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CORNELL UNIVERSITY
THE
Hlower Veterinary Cibrary
FOUNDED BY
ROSWELL P. FLOWER
for the use of the
N. Y. STATE VETERINARY COLLEGE
1897
Cc HU, Lib
QR 46.094 1897
A text-book of bacteriology, includ
iii
DATE DUE
GAYLORD PRINTEDINU.S.A.
Cornell University
Library
The original of this book is in
the Cornell University Library.
There are no known copyright restrictions in
the United States on the use of the text.
http://www.archive.org/details/cu31924000246714
A TEXT-BOOK
BACTERIOLOG
INCLUDING THE
ETIOLOGY AND PREVENTION
OF
INFECTIVE DISHASES
AND A SHORT ACCOUNT OF
YEASTS AND MOULDS, H#MATOZOA, AND
PSOROSPERMS
BY
EDGAR M. GROOKSHANK, M.B.
PROFESSOR OF COMPARATIVE PATHOLOGY AND BACTERIOLOGY, AND FELLOW OF KING'S
COLLEGE, LONDON
FOURTH EDITION
RECONSTRUCTED, REVISED AND GREATLY ENLARGED
PHILADELPHIA
W. B. SAUNDERS
925 WALNUT STREET
1897
&
To
SIR JOSEPH LISTER, BART. MB, P.RS.,
WHO HAS CREATED A NEW EPOCH IN
MEDICINE AND SURGERY,
BY APPLYING A KNOWLEDGE OF ‘MICRO-ORGANISMS
TO THE TREATMENT OF DISEASE,
This ork is, with permission, Dedicated
BY THE AUTHOR
AS A
TOKEN OF ADMIRATION AND RESPECT
PREFACE
TO THE
FOURTH EDITION.
Turs book, though nominally a fourth edition, is practically
speaking a new work. The progress of Bacteriology has been
very rapid, and many new investigations have been made in
connection with the etiology, prevention and treatment of
communicable diseases. It has been necessary to reconstruct,
enlarge and thoroughly revise the text of the third edition,
and I have added twenty-six chapters.
The most. important researches conducted in bacteriological
laboratories are those relating to the contagia. In many
diseases of man and animals it has not been possible to
identify the contagium with a bacterium, or indeed with any
micro-organism ; but when the virus is chemically examined,
or investigated with a view to protective inoculation, or utilised
for experiments in serum-therapeutics, such researches are
within the province of the bacteriologist.
The recognition of the fact that in so many diseases the
nature of the contagium has not yet been determined will
have the effect of encouraging continued activity in this
important field of scientific investigation.
I hope that this work will continue to be of use as a text-
book for the bacteriological laboratory, and that the chapters
on the etiology and prevention of the communicable diseases
viii PREFACE TO THE FOURTH EDITION.
of man and-animals will be not only of scientific interest,
but of practical value to Medical Officers of Health and
Veterinary Inspectors.
I have divided the book into three parts. Part I. is
mainly technical, and includes the most recent methods
employed in studying bacteria and investigating the etiology
of disease. Part II. deals with infective diseases and the
bacteria associated with them. Any clinical or pathological
evidence which may help to throw light on the nature and
origin of the contagia is taken into account. The most
effectual measures for stamping out these diseases are
referred to, as they are intimately connected with a
knowledge of the life-history of micro-organisms. Part III.
contains descriptions of about five hundred bacteria. Many
are of no practical importance and of very little scientific
interest, but a text-book for the laboratory cannot be con-
sidered complete unless an account is given of all bacteria
which have been more or less completely investigated. I
have endeavoured to refer to the original descriptions and
to verify them by comparison with actual cultivations, but
in a very great number of instances this has been quite
impossible, and I desire to acknowledge the assistance I have
received from the works of several authors, especially those
of Fliigge, Frankel, Hisenberg, Baumgarten, Frankland,
Sternberg, Lehmann and Neumann.
I have rearranged the bibliography according to the
chapters, and the names of authors are given in alphabetical
order. With the aid of the current numbers of the Annales
de UInstitut Pasteur, the Zeitschrift fir Hygiene, the
Centralblatt fiir Bakteriologie und Parasitenkunde, and the
Journal of Comparative Pathology and Bacteriology, it is
possible to become acquainted with the most recent litera-
ture of the subject.
Many of the coloured plates illustrating the last edition
have not been reproduced. Those substituted for them have
been drawn from my own preparations, and most of them
PREFACE TO THE FOURTH EDITION. 1x
have already appeared in my Reports to the Board of
Agriculture and in other publications.
One hundred and thirty-three woodcuts and photoraphe
have been added in the text, and I have reverted to the plan
which J adopted in the second edition, of having many of
them printed in .colours.
I take this opportunity of thanking Professor Frankel for
kindly permitting me to reproduce some of the photographs
in his excellent Atlas.
I am particularly indebted to Professor Hamilton for
the use of clich’s of figures in his classical treatise on
Pathology, and to the New Sydenham Society for several
from the’ English translation of Professor Fliigge’s well-
known work on micro-organisms.
To my Demonstrator, Dr. George Newman, D.P.H., I am
indebted for much assistance in correcting the proof-sheets,
and for the preparation of an index.
EDGAR M. CROOKSHANK.
SAInrt Hint, HAST GRINSTEAD, SUSSEX,
August 1st, 1896.
P.S.—Since this work was finally passed for press the con-
clusions of the Royal Vaccination Commissioners have been
published. I have at the last moment added extracts in the
form of a supplementary Appendix. E. M. C.
September 18th, 1896.
CONTENTS.
PART 2.
THEORETICAL AND TECHNICAL.
CHAPTER I.
HISTORICAL INTRODUCTION
CHAPTER II.
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA
CHAPTER JTL
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA
CHAPTER IV.
CHEMICAL PRODUCTS OF BACTERIA
CHAPTER V.
IMMUNITY .
CHAPTER VI.
ANTITOXINS AND SERUM THERAPY
CHAPTER VII.
THE BACTERIOLOGICAL MICROSCOPE
CHAPTER VIII.
MICROSCOPICAL EXAMINATION OF BACTERIA
CHAPTER IX.
PREPARATION OF NUTRIENT MEDIA AND METHODS OF CULTIVA-
TION .
PAGE
11
30
39
49
65
83
99
Xi CONTENTS.
CHAPTER X.
EXPERIMENTS UPON THE LIVING ANIMAL
CHAPTER XI.
EXAMINATION OF AIR, SOIL, AND WATER
CHAPTER XII.
PHOTOGRAPHY OF BACTERIA
PART 11.
ETIOLOGY AND PREVENTION OF INFECTIVE
DISEASES,
CHAPTER XIII.
SUPPURATION, PYAMIA, SEPTICA:MIA, ERYSIPELAS
CHAPTER XIV.
ANTHRAX .
CHAPTER XV.
QUARTER-EVIL.—MALIGNANT (EDEMA.—RAG-PICKERS’ SEPTI-
CASMIA.—SEPTICZMIA OF GUINEA-PIGS.—SEPTICEMIA OF
MICE .
CHAPTER XVI.
SEPTICZMIA OF BUFFALOES.---SEPTIC PLEURO-PNEUMONIA OF
CALVES.—SWINE FEVER.—SEPTICEMIA OF DEER.—SEPTI-
CMIA OF RABBITS.—FOWL CHOLERA.—FOWL ENTERITIS.
— DUCK CHOLERA.—GROUSE DISEASE
CHAPTER XVII.
PNEUMONIA.—INFECTIOUS PLEURO-PNEUMONIA OF CATTLE.—
INFLUENZA
CHAPTER XVIII.
ORIENTAL PLAGUE.—RELAPSING FEVER.—TYPHUS FEVER,—
YELLOW FEVER
PAGE
. 184
140
. 150
173
. 191
. 217
. 226
. 233
. 250
CONTENTS. xiii
PAGE
CHAPTER XIX.
SCARLET FEVER.—MEASLES ‘ 4 : ' . 261
CHAPTER XX.
SMALL-POX.—CATTLE PLAGUE. 284
CHAPTER XXI.
SHEEP-POX.—FOOT-AND-MOUTH DISEASE : 297
CHAPTER XXII.
HORSE-POX.—COW-POX z : ; 3 303
CHAPTER XXIII.
DIPHTHERIA P : : ‘ 330
CHAPTER XXIV.
TYPHOID FEVER. . 3 340
CHAPTER XXV.
SWINE FEVER. : . ‘ : . 347
CHAPTER XXVI.
SWINE MEASLES.—DISTEMPER IN DOGS.—EPIDEMIC DISEASE OF
FERRETS.—EPIDEMIC DISEASE OF MICE . , . 855
CHAPTER XXVII.
ASIATIC CHOLERA.—CHOLERA NOSTRAS.—CHOLERAIC DIARRH@A
FROM MEAT POISONING.— DYSENTERY.—CHOLERAIC DIAR-
RHGA IN FOWLS ; : ; ; : . 360
CHAPTER XXVIII.
TUBERCULOSIS : ; : ; : . 375
CHAPTER XXIX.
LEPROSY.—SYPHILIS.—RHINOSCLEROMA.—TRACHOMA 406
CHAPTER XXX.
ACTINOMYCOSIS.—MADURA DISEASE é 413
CHAPTER XXXI.
GLANDERS . j : : : 451
xiv CONTENTS.
PAGE
CHAPTER XXXII.
TETANUS.—RABIES.— LOUPING-ILL . : . . 457
CHAPTER XXXIII.
FOOT-ROT . “ . = . 2 : : ‘ . 464
CHAPTER XXXIV.
FOUL-BROOD.——INFECTIOUS DISEASE OF BEES IN ITALY.—
PEBRINE.—FLACHERIE,—INFECTIOUS DISEASE> OF CATER-
PILLARS. : ; ‘ i i : : . 469
PARE Ji.
SYSTEMATIC AND DESCRIPTIVE.
CHAPTER XXXvV.
CLASSIFICATION AND DESCRIPTION OF SPECIES. : . 475
APPENDICES.
APPENDIX I.
YEASTS AND MOULDS : . : . : . . 577
APPENDIX II.
HEMATOZOA IN MAN, BIRDS, AND TURTLES.—H#MATOZOA IN
EQUINES, CAMELS: AND FISH.—H#MATOZOA IN FROGS . 589
APPENDIX III.
PSOROSPERMS OR COCCIDIA.—AMCBA COLI . : . 609
APPENDIX IV.
APPARATUS, MATERIAL AND REAGENTS EMPLOYED IN A BAC-
TERIOLOGICAL LABORATORY . ; : : . 612
APPENDIX YV.
BIBLIOGRAPHY . : 3 . 639
SUPPLEMENTARY APPENDIX.
EXTRACTS FROM THE FINAL REPORT OF THE ROYAL VACCINA-
TION COMMISSION , : ; : ; : . 667
Dore NES
LIST OF ILLUSTRATIONS.
WOOD ENGRAVINGS AND PHOTOGRAPHS.
PAGE
. Ascococcus Billrothii, x 65 (Cohn) ; 14
. Spirocheta from Sewage Water, x 1200 (E.M. 0. ) 15
. Flagella (Koch, Brefeld, Warming, Zopf) 16
. Bacillus Megatherium (De Bary) . _ 17
. Clostridium Butyricum, x 1020 (Prazmowski) . 18
. Leuconostoc: Mesenteroides; Cocci-chains with Arthroupores (van
Tieghem and Cienkowski) ‘ 19
. Spore-bearing Threads of Bacillus Anthracis, double: stained with
Fuchsine and Methylene Blue, x 1200 (H.M.C.) 20
. Bacilli of Tubercle in Sputum, x 2500 (E.M.C.) : 21
. Comma, Bacilli in Sewage Water, stained with Gentian Violet, x 1200
(E.M.C.) 22
. Vibrios in Water eontaminated with Sewage, x 1200 a M.C.) 22
. Refraction of Light (Carpenter) » 66
. Spherical Aberration (Carpenter) 87
. Combination of Lenses in Abbé’s Homogeneous Immersion (Car-
penter) . ' 67
. Chromatic Aberration (Carpenter) 68
. Objective with Collar Correction (Zeiss) 3 69
. Microscope—English Model (Swift) 71
. Removable Mechanical Stage (Swift) 72
. Microscope—Continental Model (Zeiss) . 73
. Iris Diaphragm (Zeiss) , 74
. Abbé’s Condenser (Zeiss) . 15
. Microscope Lamp (Baker) . 6
. Large Microscope Lamp (Swift) . 5 GT
. Arrangement of Powell and Lealand’s Microscope in working ditectly
on the Edge of the Flame, with Stand for Micrometer Eye-piece to
secure Steadiness and Accuracy of! ie aa (Carpenter after
Nelson) . ‘ F 79
. Ramsden Micrometer Eye- -piece (Switt) : 80
. Micrometer Eye-piece (Zeiss) 81
. Inoculating Needles (E.M.C.) . 84
. Freezing Microtome (Swift) 94
. Microtome (Jung) . 95
. Wire-cage for Test-tubes (Muencke) ‘ 100
. Hot-air Steriliser (E.M.C.) . 101
Xvi . LIST OF ILLUSTRATIONS,
FIG. PAGE
31. Hot-water Filtering Apparatus (Muencke) 102
32. Method of making a Folded Filter (E.M.C.) 103
33, Steam Steriliser (Baird and Tatlock) . 103
34. Incubator (Muencke) 104
35. Method of Inoculating a Test-tube containing Sterile Nutrient J elly
(E.M.C.) : ; . 105
36. Levelling Apparatus (E.M.C) . 107
37. Iron box for Glass Plates (Muencke) 108
38. Method of Inoculating Test-tubes in the Preparation of Plate-cultiva-
tions (E.M.C.) . 108
39. Damp-chamber containing Plate- cultivations (Bl M.C) 110
40. Pasteur’s Large Incubator (Becker) 111
41. Petri’s Dish (Becker) : 112
42. Glass Benches and Slides (Becker) 112
43, Koch’s Serum Steriliser (Muencke) 3 114
44. Hueppe’s Serum Inspissator (Baird and Tatlock) 115
45. Box for Sterilising Instruments (Becker) ‘ 116
46. Damp Chamber for Potato-cultivations (E.M.C.) 117
47. Apparatus for Sterilisation by Steam under pressure (Baird nd
Tatlock) F E : 119
48. Drop Cultivation (Bliigge) é 121
49. Simple Method of forming a Moist Cell (Schifer) : 122
50. Warm Stage (Schafer) . E 123
51. Warm Stage shown in eciatien: (Schafer) 123
52. Warming Apparatus in Operation (Israel) 124
53. Section of Warming Apparatus and Drup-culture Slide (Israel) 125
54. Israel’s Warming Apparatus 125
55. Gas Chamber in use with Apparatus tor generating Carbonic Acid
(Schafer) : 126
56. Gas Chamber (Schafer) 126
57. Moist Cell adapted for ‘Teen antsetou of Rlsetiietty (Schafer) . 127
58. Apparatus arranged for Transmitting Electricity (Schifer) 127
59. Slide with Gold-leaf Electrodes (Schifer) p 128
60. Lister’s Flask (Becker) . 128
61, Sternberg’s Bulb (Becker) 128
62, Aitken’s Tube (Becker) 129
63. Miquel’s Bulb (Becker) . 129
64. Pasteur’s Flask (Baird and Tatlock) 130
65. Pasteur’s Double Tube (Baird and Tatlock) 130
66. Frankel’s Anaerobic Tube-culture (Frankland) ‘ 131
67. Anaerobic Culture Tube (Liborius) 132
68. Apparatus for Anaerobic Cultures (Roscoe and Lunt) é 133
69. Koch’s Syringe (Baird and Tatlock) 135
70. Syringe with Asbestos Plug (Baird and Tatlock) 135
71. Hesse’s Apparatus (Muencke) . 142
72. Sedgwick and Tucker’s Tube (Baird and Tatlock) 143
73, Pouchet’s Aeroscope (Hamilton) . 143
74. Apparatus for Estimating the number -{ Colonies in a Plate-cultiva-
tion (Muencke) 146
75. Esmarch’s Roll-culture (Frankland) 147
76. Apparatus for Counting Colonies in a Roll-culture (Becker) 148
‘
. Horizontal Micro-photographic Apparatus (Swift) 156
115.
116.
LIST OF ILLUSTRATIONS,
. Reversible Micro-photographic Apparatus (E.M.C.) , q
. Reversible Micro-photographic: Apparatus arranged in the Vertical
Position (E.M.C.) : . .
. Large Micro-photographic Apraracus (Swift) .
. Photograph of an Impression Preparation (E.M.C.) :
. Photograph of a Cultivation of Bacillus anthracis (E.M.C.) .
. Suppuration of Subcutaneous Tissue (Cornil and Ranvier)
. Pus with Staphylococci, x 800 (Fliigge)
. Subcutaneous Tissue of a Rabbit forty-eight hours sitter an Injection
of Staphylococci, x 950 (Baumgarten)
. Ulcerative Endocarditis: Section of Cardiac ‘Nidadlo, x 700 Koch)
. Pure-cultures of Streptococcus Pyogenes (E.M.C.) .
. Section of Skin in Erysipelas (Cornil and Ranvier) 2
. Streptococcus Pyogenes ke ni, Pure-cultures on Nutelent Gela-
tine (E.M.C.) .
. Streptococcus Pyogenes Bovis ; “Dies ealtanes: on Notvient Gelatine,
(B.M.C.) . : 4 ? :
|. Gonococcus, x 800 (Gunn) ‘
. Bacillus Anthracis, x 1200. Blood Corpiuctas and Bacilli unstained;
from an Inoculated Mouse (Frankel and Pfeiffer)
. Pure-cultivation of Bacillus anthracis in Nutrient Gelatine (E. M.C. )
Colonies of Bacillus anthracis, x 86 (Fltgge)
. Impression-preparation of a Colony, x 70 ee
. Margin of a Colony, x 250 (E.M.C.)
. Filaments with Oval and Irregular Elements, x 800 (B. M.C. )
. Spores of Bacillus anthracis stained with Gentian Violet, x 1500
(E.M.C.)
. Anthrax in Swine (E.M. C)
. Anthrax in Swine (E.M.C.) "
. Bacilli of Quarter-evil, x 1000 CFviinkeel and Pfeiffer)
. Pure-culture of Bacilli of Quarter-evil in Grape-sugar Caletine
(Frinkel and Pfeiffer) .
. Bacilli of Malignant &dema, x "950 {Batimgarton
. Pure-culture of Bacillus of Malignant (idema in Grape ee
Gelatine (Frankel and Pfeiffer)
. Bacilli of Malignant (Edema, x 1000 (Finkel and Pfeiffer)
. Pure-cultivation of the Bacillus of Septiceemia of Mice in Nutrient
Gelatine (E.M.C.)
. Bacterium of Rabbit Scimenats Blood of Spare, x 700 (oats
. Bacterium of Fowl-cholera, x 1200 (E.M.C.) . we A
. Bacterium of Fowl-cholera, x 2500 (E.M.C.) . :
. Bacterium of Fowl-cholera; Section from Liver of Fowl, x 700
(Fliigge)
Bacillus of Humoirhepia depicts x 950 (pauigniteny «
. Bacillus of Hemorrhagic Septicemia: Pure-culture in Gelatine
(Baumgarten) .
3. Bacterium Pneumonize Chagnon from Pleural Cavity of a "Mouse,
x 1500 (Zopf) .
. Friedlander’s Pneumococcus ; ‘Pare-caleate a Nutrient Galatians
(Baumgarten) . : j
Capsule Cocci from Pneumonia, x 1500 (Bammesrten)
Micrococcus of Sputum Septicemia, x 10C0 (Friinkel and Pfeiffer) .
b
XVii
PAGE
157
158
160
162
168
174
177
177
183
184
185
187
188
190
192
193
194
194
195
195
197
203
205
218
218
221
222
223
225
228
228
228
229
231
231
234
234
235
236
Xvili LIST OF ILLUSTRATIONS.
FIG,
117.
118.
119.
120.
121.
122.
123.
124,
125,
126. F
127.
Colonies of Sternberg’s Micrococcus, x 100 (Frankel and Pfeiffer) .
Acute Catarrhal Pneumonia, x 480 (Hamilton)
Infectious Pleuro-pneumonia of Cattle (Hamilton)
Infectious Pleuro-pneumonia of Cattle (Hamilton)
Bacillus of Influenza, x 1000 (Itzerott and ean
Bacillus of Influenza, x 1200 (E.M.C)
Bacilli of Plague and Phagocytes, x 800 (Aoyama) .
Spirillum Obermeieri in Blood of Monkey inoculated with Spirilla
after Removal of the Spleen (Soudakewitch)
Pure-cultivations of Streptococcus Pyogenes (E.M.C.)
‘ree Surface of Diphtheritic Larynx, x 350 (Hamilton)
Bacillus of Diphtheria; from a Cultivation on Blood Serum, x 1000
. (Frankel and Pfeiffer) .
28. Pure-cultures of Bacillus Diphtherie on Gigesvinny Gentine (B. M.C.)
9. Typhoid Fever. Teum of Adult, sali Sloughy and Infiltrated
Patches (Hamilton)
. Typhoid Bacilli from a Colony on Nuteioulk Gentine. x 1000
(Frankel and Pfeiffer) .
Typhoid Bacilli, x 950 (Baumgarten .
. Flagella of Typhoid Bacilli, x 1000 (Friinkel and Pfeiffer)
3. Colonies of the Typhoid Bacillus (Friinkel and Pfeiffer)
. Pure-culture of Typhoid Bacilli inoculated in the Depth of Nubrient
Gelatine (Baumgarten)
5. Typhoid Bacilli in a Section of eileen, x 800 ‘(Fltigge)
. Typhoid Bacilli in a Section of Intestine invading the Subarieors
and Muscular Layers, x 950 (Baumgarten) .
. Ulceration of the Intestine in a Typical Case of Siite: fever (E. M.C. )
. Klein's Bacillus of Swine-fever (No. 1)
. From a Preparation of Bronchial Mucus of mle s a favee
Bacillus (No. 2)
. Bacilli from an Aotiieial Cuiltare with Supres, Bacities No. 2 (Klein)
- Blood of Fresh Spleen of a Mouse after Inoculation with Swine-fever
Bacillus No. 2 (Klein). : : .
. Bacilli of Swine Erysipelas (Caenmapastony 3
. Blood of Pigeon inoculated with Bacilli of Swine Hevsinetas, « 600
(Schiitz)
. Pure-culture in Naldenk Gelatine of Basil Ppt Swine firysipelas
(Baumgarten) .
. Cover-glass Preparation of a Drop of Meat Takei Banaiantig a
Pure-cultivation of Comma-bacilli (Koch)
3, Arthrospores of Comma-bacilli (Hueppe)
7. Flagella of Comma-bacilli ; stained ny Loffler’s Method (Frankel
and Pfeiffer)
. Involution Forms of Cente taal. x ‘700 (Van inmengem))
. Colonies of Comma-bacilli on Nutrient Gelatine ; natural size (Koch)
. Colonies of Koch’s Comma-bacilli, x 60 (E.M.C.)
. Cover-glass Preparation from the Contents of a Cholera Tntesting,
x 600 (Koch)
. Cover-glass Preparation of Cholera Dejecta on Te Tinen, x 600
(Koch) .
. Section of the Mucous Membrane of a Chester inne. x 600
(Koch) . 2 . . . : .
363
364
LIST OF ILLUSTRATIONS.
. Pure-cultivations in Nutrient Gelatine of Koch’s and of Finkler’s
Comma-bacilli (E.M.C.)
. Comma-shaped Organisms with other Bacteria in Sewage- con-
. taminated Water, x 1200
. Comma,-bacilli of the Mouth,. x: 700 (Van Ermengem)
. Finkler’s Comma-bacilli, from Cholera nostras, x 700 (Fliigge)
. Deneke’s Comma-bacilli, from Cheese, x 700 (Fliigge)
. Pure-cultivation of the Spirillum Finkler-Prior, in Nutrient idlatine
(E.M.C.)
. Tropical Dysentery; Mucous Membrane of Lange Intestine (Hamilton)
. Tubercle of the Lung in a very Early Stage, x 400 (Hamilton)
. Primary Tubercle of Lung two to three weeks old, x 50 (Hamilton)
. Large Oval Giant Cell from Tubercle of Lung, x 300 (Hamilton)
. Bacillus Tuberculosis, from Tubercular Sputum, x 2500 (E.M.C.)
. Pure-cultivation of the Tubercle Bacillus on Glycerine Agar-agar
(£.M.C.)
Pure-cultivation: in Gigeetine Aipans Shae sitter ten mouths! ers
(E.M.C.)
. Pure-cultivations of Tubevele Macillne:t in a Gtyeering Aoaeapgars a sub-
culture from a Pure-culture in Glycerine Milk (E.M.C.)
. Section through a Lupus Nodule of the Nose (Hamilton)
. Tubercular Ulceration.of Mucosa of Ileum (Hamilton)
. Section of Lupus of the Skin; Giant Cell containing Tubercle
Bacillus (Fliigge)
. Tuberculosis of Pleura; “ Grapes disease” (E. M. Cc.)
. Tubercular Ulceration ‘it the Intestine of a Cow (E.M.C.)
. Tubercular Ulceration of the Intestine of a Rabbit (E.M.C.).
. Tubercular Lungs of Rabbit (E.M.C.) ‘
. Cover-glass Preparation of Pus from aChancre, x 1050 Ciastourteny
. Wandering Cell containing Bacilli (Lustgarten)
. Section of Liver from a Case of Actinomycosis in Man (E.M. ¢, )
. Actinomycotic Tumour in the Throat of a Steer (E.M.C)
. Actinomycotic Tumour of the Cheek (E.M.C.) .
. Steer with Emaciation the Result of-Actinomycosis (E.M.C. )
. Actinomycotic Growths from the. Pleura resembling ‘“ Grape-
Disease ” (E.M.C.) : z : : :
. Actinomycosis of the Skin (E.M. ©. )
. Part of Human Foot with Madura Disease (E. M. C.) .
. Bacilli of Glanders, x 700 (Fliigge)
. Section of a Branch of the Pulmonary reves sino wine Giandiors
Bacilli penctrating the Wall (Hamilton)
. Pure-culture of the Tetanus Bacillus in Grape-sugar Celatins (Frankel
and Pfeiffer)
. Foot of Sheep showing Disease of Horn (Brot)
. Section through the Foot showing a Crack extending through the
Wall
. Secreting Membrane punted wit Pungoll Girowihs (uown}
. Advanced Form of Disease of Skin between the Claws (Brown)
. Distortion of Hoof in an Advanced Form of Foot-rot (Brown)
. Diseased Comb (Cowan)
. Spores of Bacillus Alvei (E.M.C. )
. Pure-culture in Nutrient Gelatine (Cheshire anil Gheptis)
xix
PAGE.
365
366
367
367
367
370
372
376
377
377
379
380
381
381
387
388
389
390
. 393
395
396
410
410
417
424
424
425
425
430
448
452
453
458
465
465
466
466
467
469
470
470
LIST OF ILLUSTRATIONS.
. Cultivation on the Surface of Gelatine acu and one)
. Cladothrix Dichotoma (Zopf) A
. Friedlander’s Pneumococcus, x 1500 (Zopf) .
. Ascococcus Billrothii (Cohn)
. Clostridium Butyricum (Prazmowski)
. Bacillus Cyanogenus, x 650 (Neelsen)
. Pure-cultivation of Bacillus figurans on the Surface of Nutrient
Agar-agar (E.M.C.)
. Photograph of Part of an jrepieasaen Preparation of Bacillus
figurans on Nutrient Gelatine, x 50 (E.M.C.)
. Part of the same Specimen, x 200
. Bacillus Indicus : Colonies in Agar, x 60
5. Bacillus Neapolitanus, x 700 (Emmerich)
. Bacillus Megatherium (De Bary) ‘
. Pure-culture of Bacillus Megatherium in Gelatine (E, M. Cc. a
. Bacillus Putrificus Coli, x 1000 (Bienstock) . ‘ Z ‘
. Bacillus Pyogenes Feetidus, x 790 (Passet) . ‘ : 3
. Bacillus Saprogenes, No. 1 (Rosenbach)
. Bacillus Subtilis with Spores (Baumgarten)
. Pure-culture of Bacillus Subtilis in Nutrient Gelatine CBaamearten),,
. Pure-culture of Bacillus Subtilis on the Surface of Nutrient Agar
(E.M.C)
. Bacterium Zopfii ‘(Kurth) é .
5. Beggiatoa Alba (Zopf) . .
. Phase-forms of Beggiatoa Persicina (Wanniaz)
. Cladothrix Dichotoma (Zopf)
. Crenothrix Kiihniana (Zopf) . ‘
. Leuconostoc Mesenteroides (Van Tieghem and Gieckowskis.
. Micrococcus in Pyemia in Rabbits (Koch) ‘
. Proteus Mirabilis ; Swarming Islands on the Surface of Gelatine,
x 285 (Hauser)
. Proteus Mirabilis ; Involution Forms, x 624 (Hanser)
. Proteus Vulgaris, x 285 (Hauser
. Sarcina, x 600 (Fliigge)
5. Spirocheta Plicatile (E.M.C.) . ‘ :
3. Comma-bacilli in Water contaminated with Sewage .
. Comma-bacilli of the Mouth, x 700 (Van Ermengem)
. Deneke’s Comma-bacilli, from Cheese, x 700 (Fliigge)
Streptococcus in Progressive Tissue Necrosis in Mice (Koch)
. Vibrio Rugula, x 1020 (Prazmowski) .
. Black Torula; Pure-cultivation on Potate (E. M. C.) -
. Head and Neck of Calf with Advanced Ringworm (Brown)
. Non-pigmented Amceboid Forms (Marchiafava and Celli)
. Pigmented Amceboid Forms (Golgi)
. Semi-lunar Bodies of Laveran (Golgi) .
. Rosette Forms with Segmentation (Golgi)
é Hlagellated Forms (Vandyke Carter)
. “Surra” Parasites, occurring Singly and Fused, x 1200
. Parasites in the Blood of Rats (Lewis)
. A Monad in Rat’s Blood, x 3000 (E.M.C.)
. Monads in Rat’s Blood, x 1200 (E.M.C.)
. Monads in Rat’s Blood stained with Methyl Violet, x "1200 (E, MLC, )
FIG.
243.
244,
245,
246,
247.
248,
249.
250.
251,
252.
253.
254.
255.
256.
257.
253.
259.
260.
261.
262.
263.
264.
265.
266.
267.
268.
269.
270.
971.
272.
273,
LIST OF ILLUSTRATIONS.
Organisms in the Blood of Mud-fish (Mitrophanow)
Organisms in the Blood of the Carp (Mitrophanow)
Ameeba cola in Intestinal Mucus (Lésch)
Warm Stage (Schiifer)
Warm Stage (Stricker) .
Combined Gas Chamber and Warm Stage (Stricker) s
Vertical Micro-photographic Apparatus (Leitz)
Koch's Steam Steriliser (Muencke)
Hot-air Steriliser (Muencke) .
Section of Hot-air Steriliser (Muencke)
Hot-water Filtering Apparatus with Ring Tints Chohibecks
Wire-cage for Test-tubes (Muencke)
Platinum-needles (E.M.C.)
Damp Chamber for Plate-cultivations (E. M.C. )
Apparatus for Plate-cultivations (E.M.C.)
Box for Glass-plates (Muencke)
Glass Benches for Glass-plates (Becker)
Israel’s Case (Becker)
-Damp Chamber for Potato CaNawations (E.M. 0. si
Koch’s Serum Steriliser (Muencke)
Serum Inspissator (Muencke) . : ,
D’Arsonval’s Incubator (Muencke) ’
Schlosing’s Membrane Regulator (Mioenékey «
Gas Burner protected with Mica Cylinder (Muenckey
Koch’s Safety Burner (Mueéncke)
Babés’ Incubator (Muencke)
Moitessier’s Gas-pressure Regulator (Muencke)
Reichert’s Thermo-regulator (Muencke)
Meyer’s Thermo-regulator (Muencke) .
Siphon Bottle with Flexible Tube, Glass Nozle, anda : Mohr's 's Tings
cock (E.M.C.) . 2 : : 4 '
Desiccator (E.M.C.)
DESCRIPTION OF PLATES.
DESCRIPTION OF PLATE I.
Bacteria, Schizomycetes, or Fission Fungi.
Following p. 14.
1. Cocci singly and varying in size. 2. Cocci in chains or rosaries (strepto-
coccus). 3. Cocci in a mass (staphylococcus), 4 and 5. Cocci in pairs
(diplococcus). 6. Cocci in groups of four (merismopedia). 7. Cocci in packets
(sarcina). 8. Bacterium termo. 9. Bacterium termo x 4000 (Dallinger and
Drysdale). 10. Bacterium septicemia hemorrhagice. 11. Bacterium pneu-
monie croupose. 12. Bacillus subtilis. 13. Bacillus muriseptieus. 14.
Bacillus diphtheria. 15. Bacillus typhosus (Eberth). 16. Spirillum undula
(Cohn). 17%. Spirillum volutans (Cohn). 18. Spirillum cholere Asiatice.
19. Spirillum Obermeiert (Koch). 20. Spirocheta plicatilis (Fliigge). 21.
Vibrio rugula (Prazmowski). 22. Cladothrix Forsteri (Cohn). 23. Cladothrix
dichotoma (Cohn), 24. Monas Okenii (Cobn). 25. Monas Warmingii (Cobn).
26. Rhabdomonas rosea (Cohn). 27. Spore-formation (Bacillus alvei). 28.
Spore-formation (Bacillus anthracis). 29. Spore-formation in bacilli cultivated
from a rotten melon (Frinkel and Pfeiffer). 30. Spore-formation in bacilli
cultivated from earth (Frankeland Pfeiffer). 31. Involution-form of Crenothriz
(Zopf). 32. Involution-forms of Vibrio serpens (Warming). 33. Involution-
forms of Vibrio rugula.(Warming). 34. Involution-forms of Clostridium
polymyxa (after Prazmowski). 35. Involution-forms of Spirillum cholere
Asiatice. 36. Involution-forms of Bacterium aceti (Zopf and Hansen).
37. Spirulina-form of Beggiatoa alba (Zopf). 38. Various thread-forms of
Bacterium merismopedioides (Zopf). 39. False-branching of Cladothria (Zopf).
DESCRIPTION OF PLATE II.
Pure-cultivations of Bacteria.
Following p. 100.
Fie. 1.—In the depth of Nutrient Gelatine. A pure-cultivation of Koch’s
comma-bacillus (Spirillum cholere Asiatice) skowing in the track of
the needle a funnel-shaped area of liquefaction enclosing an air-bubble,
and a white thread. Similar appearances are produced in cultivations of
the comma-bacillus of Metchnikoff.
Fig. 2.—On the surface of Nutrient Gelatine. A pure-cultivation of Bacillus
typhosus on the surface of cbliquely solidified nutrient gelatine.
xxii
DESCRIPTION OF PLATES. xxili
Fig. 3.—On the surface of Nutrient Agar-agar. Pure-cultivation of Bacillus
indicus on the surface of obliquely solidified nutrient agar-agar. The
growth has the colour of red sealing-wax, and a peculiar crinkled
appearance. After some days it loses its bright colour and becomes
purplish, like an old cultivation of Micrococcus prodigiosus.
Fig. 4.—On the surface of Nutrient Agar-agar. A pure-cultivation obtained
from an abscess (Staphylococcus pyogenes aureus). ;
Fie. 5.—On the surface of Nutrient Agar-agar. A pure-cultivation obtained
- from green pus (Bacillus pyocyaneus). The growth forms a whitish,
transparent layer, composed of slender bacilli, and the green pigment
is diffused throughout the nutrient jelly. The growth appears green by
transmitted light, owing to the colour of the jelly behind it.
¥ie. 6.—-On the surface of Potato. A pure-cultivation of the bacillus of
glanders on the surface of sterilised potato.
DESCRIPTION OF PLATE III.
Plate-cultivation.
Following p. 108.
This represents the appearance of a plate-cultivation of the comma-bacillus
of Cholera nostras, when it is examined over a slab of blackened plate-glass.
The drawing was made from a typical result of thinning out the colonies by
the ane of plate-cultivation. At this stage they were completely isolated
one from the other; but later they became confluent, and produced complete
liquefaction of the gelatine.
DESCRIPTION OF PLATE IV.
Streptococcus Pyogenes.
Following p. 178.
Fig. 1—From a cover-glass preparation of pus from a pyzmic abscess.
‘Stained with gentian-violet by the method of Gram, and contrast-stained
with eosin. x 1200. Powell and Lealand’s apochromatic ~, Hom. imm.
E. P. 10.
Fig. 2.—From cover-glass preparations of artificial cultivations of the strepto-
coccus in broth and in milk at different stages of growth. x 1200. Powell
and Lealand's apochromatic 7; Hom. imm. E. P. 10.
In these preparations there is a great diversity in size and form of the
chains and their component elements. In the drawing examples are
figured of the following:
(a) Branched chains.
(b) Simple chains composed of elements much smaller than the
average size.
(ce) Chains with spherical and spindle-shaped elements at irregular
intervals. These are conspicuous by their size, and are sometimes
terminal.
(d e) Chains in which the elements are more or less uniform in size.
(f) Complex chains with elements dividing both longitudinally and
transversely, and varying considerably in size in different lengths
of the same chain.
‘
xxiv DESCRIPTION OF PLATES.
DESCRIPTION OF PLATE V.
Bacillus Anthracis.
Following p. 192. N
¥1g.'1.—From a cover-glass preparation of blood from the spleen of a guinea-
pig inoculated with blood from a sow. x 1200. Powell and Lealand’s
apochromatic 7; Hom.imm. E, P. 10.
Fig. 2.—From a section of a kidney of a mouse. Under a low power the
preparation has exactly the appearance of an injected specimen. Under
higher amplification the bacilli are seen to have threaded their way along
the capillaries between the tubules, and to have collected in masses in
the glomeruli. Stained with Gram’s method (gentian-violet), and eosin.
x 500. :
Fig. 3.— Bacillus anthracis and Micrococcus tetragenus. From a section from
the lungs of a mouse which had been inoculated with anthrax three days
after inoculation with Micrococcus tetragenus. A double or mixed infection
resulted. Anthrax-bacilli occurred in vast numbers, completely filling the
small vessels and capillaries, and in addition there were great numbers
of tetrads. Stained by Gram’s method (gentian-violet), and with eosin.
x 500.
DESCRIPTION OF PLATE VI.
Bacillus Murisepticus.
Following p. 224.
Fig. 1.—From a section of a kidney of a mouse which had died after inocula
tion with a pure-cultivation of the bacillus. With moderate amplification,
the white blood-corpuscles have a granular appearance, and irregular
granular masses are scattered between the kidney tubules. Stained by
Gram’s method with eosin. x 200.
FIG. 2.—Part of the same preparation with high amplification. The granular
appearances are found to be due to the presence of great numbers of
; extremely minute bacilli. x 1500. ;
DESCRIPTION OF PLATE VII.
Casual Cow-pox.
Following p. 278.
¥ig. 1—Case of W. P——,a milker, infected from the teats of a cow with
natural cow-pox. There was a large depressed vesicle with a small
central crust and a tumid margin, the whole being surrounded by a
well-marked areola and considerable surrounding induration.
Fic. 2.—The same case a week later, showing a reddish-brown crust on a
reddened elevated and indurated base.
DESCRIPTION OF PLATES. XxVv
DESCRIPTION OF PLATE VIII.
Bacillus diphtheriz and Bacillus typhosus.
Following p. 332.
Fig. 1—Cover-glass preparation from a pure-cultivation of Bacillus diph-
theriz on blood serum; obtained from the throat in a typical case of
diphtheria. Stained with gentian-violet. x 1200.
Fig. 2,—Cover-glass preparation from a pure-cultivation of Bacillus typhosus
on nutrient-agar; from the spleen in a case of typhoid fever, Stained
with gentian-violet. x 1200.
DESCRIPTION OF PLATES IX, AND X.
Swine Fever.
Following p. 348.
PLATE IX.—Part of intestine from a typical case of swine fever, showing
scattered ulcers and ulceration of the ileo-czcal valve.
PLATE X.—From the same case of swine fever. The lungs were extensively
inflamed and partly consolidated, and the lymphatic glands were enlarged
and of a deep red or reddish-purple colour. i
DESCRIPTION OF PLATE XI.
Bacillus tuberculosis.
Following p. 378.
The figures in this plate represent the bacilli of tuberculosis in
different animals, examined under the same conditions of amplifica-
tion and illumination. x 1200. Lamp-light illumination.
Fie. 1.—Bacilli in pus from the wall of a human tubercular cavity. In
this specimen the bacilli are shorter than those in tubercular sputum,
and are very markedly beaded.
Fig. 2.—Bacilli in pus from a tubercular cavity from another case in man.
They are present in the preparation in enormous numbers. The proto-
plasm occupies almost the whole of the sheath, and the bacilli are
strikingly thin and long.
Fig. 3.—Bacilli in sputum from an advanced case of phthisis, showing
the ordinary appearance of bacilli in sputum; some beaded, others
stained in their entirety; occurring both singly and in pairs, and
in groups resembling Chinese letters.
Fig. 4.—Bacilli in a section from the lung in a case of tuberculosis in man,
The bacilli in human tuberculosis are found in, and between, the tissue
cells ; and sometimes, as in equine and bovine tuberculosis, in the
interior of giant cells, but not so commonly.
Fig. 5.—From a cover-glass preparation of the deposit in a sample of milk
from a tubercular cow. The bacilli were longer than the average
length of bacilli in bovine tissue sections, and many were markedly
beaded.
XXVi DESCRIPTION OF PLATES.
Fic. 6.—¥rom a section of the brain in a case of tubercular meningitis in a
calf, showing a giant cell containing bacilli with the characters usually
found in sections of bovine tuberculosis.
Frc. 7—From a section of the liver of a pig with tubercle bacilli at the
margin of a caseous nodule.
Fra. 8.—From a cover-glass preparation of a crushed caseous mesenteric
gland from a rabbit infected by ingestion of milk from a cow with
tuberculosis of the udder,
Fig. 9.—From a section of lung in a case of equine tuberculosis, showing a
giant cell crowded with tubercle bacilli.
Fiq..10.—From a section of lung from a case of tuberculosis in the cat, with ,
very numerous tubercle bacilli.
Fig. 11.—From a cover-glass preparation of a crushed caseous nodule from
the liver of a fowl, with masses of bacilli. These are for the most part
short, straight rods; but other forms, varying from long rods to mere
granules, are also found.
Fie. 12.—From sections of the liver and of the lung in a case of tubercu-
losis of a Rhea, Isolated bacilli are found, as well as bacilli packed in
large cells, colonies of sinuous bacilli, and very long forms with terminal
spore-like bodies and free oval grains,
The preparations from which these figures were drawn were all
stained by the Ziehl-Neelsen method, with the exception of the first,
which was stained by Ehrlich’s method.
DESCRIPTION OF PLATE XIL
Tubercular Mammitis.
Following p. 394.
Fig. 1.—From a section of the udder of a milch cow. The tubercular deposit
is seen to invade the lobules of the gland. Lobules comparatively healthy
are marked off, more or less sharply, from the diseased ones in which the
new growth in its progress compresses and obliterates the alveoli. Stained
by the Ziehl-Neelsen method and with methylene-blue. x 50.
Fia. 2.—Part of the same preparation. On the right of the section part of a
healthy lobule is seen. On the left a lobule is invaded by tubercular new
growth composed of round cells, epithelioid cells and typical giant cells.
Tubercle bacilli can be seen both singly and collected in groups. They
are found in and between the cells, and in the interior of giant cells.
Bacilli may be seen between the cells lining an alveolus and projecting
into its lumen. x 800.
DESCRIPTION OF PLATE XIII.
Tuberculosis in Swine.
Following p. 400.
Section of liver of a pig with scattered tubercular nodules. Microscopical
sections of the liver showed tubercle bacilli in very small numbers,
DESCRIPTION OF PLATES. XXVii
DESCRIPTION OF PLATE XIV.
Bacillus Lepra. ©
Following p. 408.
Fre. 1.—From a section of the skin of a leper. The section is, almost in
its entirety, stained red, and, with moderate amplification, has a finely
granular appearance. Stained by the Ziehl-Neelsen method (catbblised
fuchsine and methylene-blue). x 200.
Fiq. 2.—Part of the same preparation with bigh amplification, showing that
the appearances described above are due entirely to an invasion of the
tissue by the bacilli of leprosy. x 1500.
DESCRIPTION OF PLATES XV. AND XVI.
Actinomyces.
Following p. 432.
PLATE XV,
Fic. 1.—From a preparation of the grains from an actinomycotic abscess in
a boy; examined in glycerine. The drawing has been made of a com-
plete rosette examined by focussing successively the central and peripheral
portions. Towards the centre the extremities of the clubs are alone
visible; they vary in size, and if pressed upon by the cover-glass give the
appearance of an irregular mosaic. Towards the periphery the clubs are
seen in profile, and their characteristic form recognised. At one part
there are several elongated elements, composed of separate links. x 1200.
Fig. 2.—Different forms of clubs from preparations in which the rosettes have
been flattened out by gentle pressure on the cover-glass. x 2500.
(a) Single club. (4) Bifid club. (¢) Club giving rise to four
secondary clubs. (d) Four clubs connected together, recalling
the form of a bunch of bananas. (¢) Mature club with a lateral
bud. (f) Apparently a further development of the condition
represented at (e). (g) Club with a lateral bud and transverse
segmentation. (h) Single club with double tranverse segmenta-
tion. (é) Club with oblique segmentation. (j) Collection of
four clubs, one with lateral gemmation, another with oblique
segmentation, (%) Club with lateral buds on both sides, and
cut off square at the extremity. (2) Club with a daughter club
which bears at its extremity two still smaller clubs. (m) Club
divided by transverse segmentation into four distinct elements,
(n) Elongated club composed of several distinct elements. (0) and
(~) Clubs with terminal gemmation. (q) Palmate group of clubs.
(r) Trilobed club. (s) Club with apparently a central channel.
(t) Filament bearing terminally a highly refractive oval body.
PLATE XVI.
Fic. 1.—From a section of a portion of the growth removed from a boy
during life. The tissue was hardened in alcohol, and cut in celloidin.
The section was stained by Gram’s method and with orange-rubin. x 50.
Fic. 2,—From the same section. A mass of extremely fine filaments occupies
the central part of the rosette. Many of the filaments have a terminal
enlargement. The marginal part shows a palisade of clubs stained by the
orange-rubin. x 500.
Xxvili DESCRIPTION OF PLATES.
Fes. 3 and 4.—From cover-glass preparations of the fungus teased out of the
new growths produced by inoculation of a calf with pus from a boy
suffering from pulmonary actinomycosis. Stained by Gram’s method and
orange-rubin. The threads are stained blue and the clubs crimson (@)
In the younger clubs the thread can be traced into the interior of the
club (4). In some of the older clubs the central portion takes a yellowish
stain, and in others the protoplasm is not continued as a thread, but is
collected into a spherical or ovoid or pear-shaped mass. In others, again,’
irregular grains stained blue are scattered throughout the central portion
(Fig. 4). x 1200.
Fig. 5.—From a pure-culture on glycerine-agar. (@) branching filaments,
(b) a mass of entangled filaments. Gram’s method. x 1200.
Fig. 6.—From a similar but older cultivation. (a) a filament with spores,
(b) chains of spores simulating streptococci. Gram’s method. » 1200.
DESCRIPTION OF PLATES XVII. AND XVIII.
Actinomycosis Bovis.
Following p. 434.
PLATE XVII.
Section of an actinomycotic tongue stained by the method of
Gram and with eosin.
Fig. 1.—This illustrates the appearance which is usually seen under a low
power, when a section is stained by Gram’s method and with eosin. The
central portion of a mass of the fungus is either unstained or tinged with
eosin, while the marginal portion is stained blue. The reverse is seen, as a
rule, in sections from man ; although under a low power the general appear-
ance of sections from these two sources is somewhat similar. x 50.
Fig. 2.—a, b, c, d, represent the earliest recognisable forms of the ray fungus
in the interior of leucocytes. In e the club-forms can be recognised. In
f and g there are small stellate groups of clubs. x 500.
Fig. 3.—A part of the section represented in Fig. 1, under a high power. The
marginal line of blue observed under a low power is now recognised as the
result of the stain being limited to the peripherally arranged clubs. At
(a) part of a rosette has undergone calcification ; the clubs are granular,
and have not retained the stain. At (4) and close to it there are the
remains of rosettes in which the process of calcification is almost complete.
x 500. ;
Puate XVIII.
The figures in this plate are taken from sections of a case of
so-called “osteosarcoma,” in which the growth of the fungus was
remarkably luxuriant. The specimens were stained by Plauts’
method.
Fig. 1.—Different forms of clubs in different specimens: x 1200.
(a) Very small club-shaped elements.
(6) A club with transverse segmentation.
(ce) A club with lateral daughter clubs.
DESCRIPTION OF PLATES, xxix
(4 and e) Clubs with terminal offshoots resembling teleutospores.
(f) A club with developing daughter clubs on the left, and on the
right a mature secondary club.
(g) A segmental club with lateral offshoots.
(A) “Two clubs undergoing calcification.
Fig. 2.—A very remarkable stellate growth comprised of nine wedge-shaped
collections of clubs radiating from a mass of finely granular material.
x 500.
Fic. 3.—A rosette undergoing central calcification, and consisting in part of
extremely elongated clubs resembling paraphyses. Calcareous matter is
also being deposited in the club-shaped structures. x 500.
Fie, 4.—Part of a rosette with continuation of the club-shaped bodies
into transversely segmented Wepnebing cells apparently representing short
hyphe. .« 500.
Fig. 5.—A ‘rosette from another section in which similar appearances are
observed as in Fig. 4. x 500.
DESCRIPTION OF PLATE XIX.
Pure-cultivations of Actinomyces.
Following p. 488.
These tubes were selected from a great number of cultivations
in which there were different appearances. In some instances the
growths had a faint tinge of pink.
Fig. 1.—Pure-cultivation on the surface of potato, showing a luxuriant
sulphur-yellow growth entirely composed of entangled masses of fila-
ments. After three months’ growth.
Fig. 2.—Pure-culture from the same series, on glycerine-agar. In this case
the culture remained perfectly white. The jelly was coloured reddish-
brown. After fifteen months’ growth.
Fig.°3.—Pure- culture on glycerine-agar in which the growth was dark-
brown, in parts black, and the jelly stained dark-brown. After nearly
two years’ growth.
DESCRIPTION OF PLATES XX. AND XXI.
Actinomycosis Bovis.
Following p. 440.
PLATE XX.
Fie. 1.—From a section of an actinomycotic tongue stained by the triple
method (Ziehl-Neelsen, logwood and orange-rubin). In this section the
separate centres of growth are clearly shown. Each neoplasm consists of
a fungus system, in which the masses of the fungus, situated more or less
centrally, are surrounded with round cells, epithelioid cells, sometimes
giant cells, and lastly fibrous tissue forming a more or less distinct
capsule. In parts the fungi have fallen out of the section. x 50.
Fie. 2.—From a section of a ‘‘tubercular” nodule from the lungs of a
Norfolk heifer with pulmonary actinomycosis. The nodule is a multiple
growth surrounding a bronchus, and is enclosed by a capsule, in the
XXX DESCRIPTION OF PLATES.
vicinity of which the pulmonary alveoli are compressed. It is composed
of a number of separate neoplasms, and each of the latter is composed of
secondary centres of growth resembling the giant-cell systems of bacillary
tuberculosis. The new growth is composed of ray-fungi, large multi-
nucleated cells, sometimes distinct giant cells, round cells, epithelioid cells,
and, surrounding them, fibrous tissue. On examination of the same
specimen with a higher power the typical rosettes of clubs are sometimes
surrounded by multinucleated cells, and sometimes small rosettes are
found like tubercle bacilli, in the interior of giant cells. From a pre-
paration stained by Ziehl-Neelsen, logwood, and orange-rubin. x 50.
PLATE XXI.
Fie. 1.—(a@) A leucocyte containing the fungus in its earliest recognisable
form. (%) A large multinucleated cell containing the fungus in an early
stage with the club-form already visible. (¢) A leucocyte containing a
small stellate fungus. (d@) A large cell containing clubs arranged in a
small rosette. (¢) A multinucleated cell with clubs arranged in a palmate
form. All the above are drawn from sections of actinomycotic tongues
stained by the triple method. x 500.
Fig. 2.—A giant cell with large vesicular nuclei at the periphery, and in the
centre a fully formed rosette of actinomyces with a smaller growth within
a “daughter” cell. From a section of the tongue of an ox stained by
the triple method. x 500.
Fic. 3.—A very large circular giant cell, with its ring of nuclei at the
periphery, enclosing several isolated tufts of actinomyces. From a section
of a nodule in the lung. Stained by the triple method. x 500.
Fic. 4.—Three rosettes of actinomyces surrounded by a row of large, some-
what angular multinucleated cells. From a section of the tongue of an
ox stained by the triple method. x 430. :
DESCRIPTION OF PLATE XXII.
Bacillus tetani.
Following p. 458,
Fic, 1.—From a cover-glass preparation of a pure-cultivation of the tetanus
bacillus in broth; stained with Neelsen’s carbolised fuchsine. » 1200.
Lamplight illumination.
Fre, 2.—From a cover-glass preparation from the same source; stained with’
Neelsen’s solution and methylene blue. x 1200. Lamplight illumination.
PART I.
THEORETICAL AND TECHNICAL.
BACTERIOLOGY:
AND
INFECTIVE DISEASES.
CHAPTER I.
HISTORICAL INTRODUCTION.
Tue researches of Pasteur into the réle played by bacteria in the
processes of fermentation and putrefaction, and the investigations of
‘the practical mind of Lister, with the resulting evolution of antiseptic
surgery, demonstrated the necessity for a more intimate acquaint-
ance with the life-history of these micro-organisms. Further re-
searches in diseases such as anthrax, the silkworm malady, pyemia,
septicemia, and fowl-cholera, invested the science of Bacteriology
with universal interest and vast importance; while the investiga-
tions which established an intimate connection between bacteria
and other infective diseases, and more especially the discovery by
Koch of bacteria in tuberculosis and in Asiatic cholera, claimed the
attention of the whole thinking world.
Those bacteria which are connected with disease, and. more
especially those which have been proved to be the causa cwusans, are
of predominant interest and importance.
The first attempt to demonstrate the existence of a contagium
vivum dates back almost to the discovery of the microscope.
Athanasius Kircher, nearly two and a half centuries ago, expressed
his belief that there were definite micro-organisms to which diseases
were attributable. The microscope had revealed that all decom-
posing substances swarmed with countless micro-organisms which
were invisible to the naked eye, and Kircher sought for similar
organisms in diseases which he considered might be due to their
agency. The microscope which he described obviously could not
1
2 BACTERIOLOGY.
admit of the possibility of studying, or even detecting, the micro-
organisms which are now known to be associated with certain
diseases ; and it is not surprising that his teachings did not at the
time gain much attention. They were destined, however, to receive
a great impetus from the discoveries which emanated from ‘the
father of microscopy.”
Antony van Leeuwenhoek had learned as 4 youth to grind and
polish lenses, and later in life employed his spare time in constructing
microscopes, and in conducting those researches which have made
for him a name which is familiar to all microscopists. His researches
were published in a series of letters to the Royal Society. In 1675
he described extremely minute organisms in rain-water, well-water,
infusions of pepper, hay, and other vegetable and animal substances,
in saliva, and in scrapings from the teeth; and, further, he was
able to differentiate these minute living things by their size, their
form, and the character of their movements. In 1683 these
discoveries were illustrated by means of woodcuts, and there can be
little doubt, from the drawings of these micro-organisms, that they
are intended to represent leptothrix filaments, vibrios, and spirilla.
Indeed, we can almost recognise these micro-organisms as bacteria
from Leeuwenhoek’s graphic descriptions, apart from his figures.
They were described as moving in the most characteristic manner,
progressing with great rapidity, or spinning round like a top, and
so excessively minute that they were only perceived with great
difficulty. The smallest forms could hardly be examined individually ;
but, viewed en masse, they closely resembled a swarm of gnats or
flies. In another communication, published in 1692, he gives
some idea of the size of these animalcules by stating that they
were a thousand times smaller than a grain of sand. Others
which were, comparatively speaking, of considerable length, were
characterised by their peculiar mode of progression, bending and
rolling on themselves—movements which, he adds, created both
delight and astonishment in the mind of the observer. Leeuwenhoek
himself was not disposed to believe in the possibility of such
organisms being found in the blood in disease; but as soon as he
had proved the actual existence of such minute creatures, theoretical
physicians were not wanting who at once attributed various maladies
to their agency. Among these, Nicholas Andry is made conspicuous
by his work published in 1701, Andry classed the minute organisms
discovered by Leeuwenhoek as worms.
In 1718 Lancisi believed that the deleterious effect. of the air of
malarial districts depended upon animalcules, and others considered
HISTORICAL INTRODUCTION, 3
that the plague in Toulon and Marseilles in 1721 arose from a
similar cause. In fact, by some, all diseases were attributed to
vermicules, and this led to the theory being ridiculed and discredited.
In spite of adverse criticism, the theory of contagium vivum
survived, and Linnzus acknowledged it by placing the micro-
organisms discovered by Leeuwenhoek, the contagia of specific
fevers, and the causes of putrefaction and fermentation, into one
class—‘ chaos.” The theory was further supported by the writings
of Plenciz, who, in 1762, very ably discussed the nature of contagium,
as well as the relation of animalcules to putrefaction and disease.
However, no proofs in support of these theories were forthcoming,
and gradually the idea of contagiwm vivum fell into obscurity, and
indeed came to be regarded by some as an absurd hypothesis.
Though a causal relation of animalcules to diseases was for a
time discredited, the natural history of these micro-organisms was
studied with increasing interest. In 1778 Baron Gleichen described
and figured a great number of micro-organisms which he had
discovered in various vegetable infusions. Joblot, Lesser, Réaumur,
Hill, and many others worked at the same subject. Hill remarked
that there was hardly the least portion of matter or the least drop
of fluid of any kind naturally found in the earth, which was not.
inhabited by multitudes of animalcules. But these observers inclined
rather to searching for new forms than to studying more thoroughly
those which had been already discovered; and, as a result, but little
scientific progress was made until the time of Miiller, of Copen-
hagen. Miller, in 1786, criticised the work of previous writers,
and pointed out that they had been too much occupied with merely
finding new micro-organisms. Miiller took into account the form
of the micro-organism, its mode of progression, and other biological
characters, and on such data based a classification. Thus the
scientific knowledge of these minute beings was considerably advanced
by his writings and illustrations.
The subject which now eclipsed all others in interest was the
origin of these micro-organisms. Two rival theories were widely
discussed—spontaneous generation, and development from pre-exist-
ing germs; and the researches that were made in the course of
this discussion, and the discoveries which resulted, indirectly yet:
materially advanced the germ theory of disease, and explain many
of the phenomena in the life-history of the pathogenic microbes.
which have been brought to light in recent years.
Spontaneous development of micro-organisms in putrescible
infusions was believed in by many, but was supported by no one
4 BACTERIOLOGY.
with greater persistency than Needham. Needham found that
animalcules readily developed when meat infusion was boiled anp
transferred to a well-stoppered flask, and he could only explain this
by supposing that they originated spontaneously from the material of
the infusion. In 1768 Bonnet strenuously opposed these conclusions
on purely theoretical grounds, and maintained that it was far more
probable that the ova of the animalcules were present in the infusions
or were suspended in the air enclosed in the flask.
Spallanzani was the first to demonstrate by experiment the
correctness of Bonnet’s arguments. It occurred to him to boil the
infusion in flasks, and to seal the vessels during the process of boiling.
As a result the flasks remained free from putrefaction, and
animalcules only developed when the infusion was exposed to the air
by making a hole in the flask. That Spallanzani’s experiments
were reliable, and his conclusions correct, was evidenced by the fact
that his simple precaution led to great practical results, as Francois
Appert introduced, on this principle, the method of preserving meats,
vegetables, and other provisions.
The disciples of Needham nevertheless brought forward counter
objections. Treviranus urged that a certain guantity and quality of
air was necessary for the spontaneous development of these infusoria,
and that by sealing the flasks, too small a quantity of air was in
contact with the infusion, and, further, that this air had become
changed in quality by the process of boiling.
Spallanzani argued against these objections, but did not support
his opinions by further experiments, so that the question remained
for a time undecided.
In 1836 Francis Schulze devised an experiment which brought
still further evidence against Needham’s theory. Schulze filled a
glass vessel half full with distilled water and different animal and
vegetable substances. This was plugged with a doubly-bored cork,
and through each perforation a glass tube was introduced, bent at
a right angle. On boiling the flask, steam issued freely from each
tube, and all parts were thoroughly sterilised. Each tube was then
connected with a bulbed tube, one bulb containing concentrated
sulphuric acid and the other a solution of potash. Fresh air was
drawn into the flask by aspiration, and this was deprived of any
germs which might be present by its passage through the sulphuric
acid. The result was that the infusion remained without any
development of micro-organisms. When, on the other hand, air was
admitted without first being drawn through the sulphuric acid, the
infusion in a short time teemed with animalcules. In other words,
~
HISTORICAL INTRODUCTION, o
Schulze demonstrated that in spite of free access to air, which had not
been heated, the infusions remained free from germs.
Schwann, in 1837, arrived at similar results. He found that
putrescible substances remained sterile if exposed to an abundant
supply of air which was heated by being passed through a melted mix-
ture of metals. This convinced him that the cause of the decompo-
sition which would otherwise have occurred must exist in the air.
The objection remained that in the experiments of Schulze and
Schwann, the air which was admitted to the flasks had undergone
either a chemical or a thermal change, and therefore the theory of
Needham was not yet entirely disposed of.
In 1854 the final blow was dealt by Schroder and Van Dusch.
These investigators demonstrated that decomposition could be obviated
without resorting either to thermal or chemical treatment of the
air, as simple filtration of the air through cotton-wool was shown to.
be efficacious in excluding germs. Finally, Hoffman in 1860, and
independently, Chevreuil and Pasteur in 1861, showed that even
cotton-wool could be dispensed with, as a sterile solution would
remain sterile when the neck of the vessel was bent into an
S-shaped curve. Micro-organisms in the air entering the flask
were deposited by gravitation in the bend of the tube.
The advocates of spontaneous generation were ready with fresh
objections. They now urged that the medium lost its power of
undergoing decomposition by being boiled. This objection was at
once set aside by the fact that when unfiltered air was admitted to
the infusion, decomposition set in. Additional evidence was brought
against spontaneous generation by the experiments of Pasteur,
Burdon Sanderson, Lister, and others, in which it was shown that
blood, urine, and milk would remain without decomposition, when all
precautions were adopted to avoid contamination in filling the
sterilised flasks.
Even at this stage of this great scientific controversy fresh
difficulties arose, for it was found that in certain solutions which had
been boiled and hermetically sealed in flasks micro-organisms made
their appearance. In 1872 Charlton Bastian published a research
which was to prove that spontaneous generation actually took
place. Decoctions of turnip and cheese which had been filtered,
neutralised, and boiled for ten minutes, and hermetically sealed
during the boiling, were found after a time to contain micro
organisms. These results, however, were before long explained by
the fact that in milk, infusions of hay, and certain other decoctions,
the spores of bacilli are present, which are much more resistant
6 BACTERIOLOGY.
than the bacilli themselves. In such cases mere scalding or boiling
for a few minutes will not sterilise the solution. The bacilli are
destroyed, but not their spores; and if the latter remain unhurt,
they will germinate, and rapidly multiply. But if, as Tyndall
found, the boiling be repeated a second and a third time, all the
spores will be destroyed; for in the intervals between the boilings
the spores sprout into bacilli, and the bacilli at the next boiling
perish ; so that after three or four repeated boilings the infusion is
rendered perfectly free from germs.
While this discussion was occupying the attention of the whole
scientific world, some investigators had been again following up the
theory of a connection between micro-organisms and disease.
In 1837 Cagniard Latour and Schwann independently made the
discovery that the yeast plant was a living organism, and the true
cause of yeast fermentation. The close analogy between the pro-
cesses of fermentation and of certain diseases had long been held;
and, therefore, when it was proved that fermentation was due to a
micro-organism, fresh advocates appeared in support of the theory
that diseases were produced by similar agencies. Boehm, in 1838,
described certain organisms in cholera, which was at that time
raging in Europe; but the researches of Bassi, who a year
previously had discovered the cause of a disease of silkworms,
attracted much greater attention.
Bassi discovered that in this disease extremely minute spores
existed on the bodies of the worms, which were conveyed from the
sick to the healthy. They destroyed the healthy worms by
germinating in their skins and growing into their bodies. These
discoveries may be said to have brought the theory of contagium
vivum to life again; and Henle, in reviewing the facts of the case in
1840, came to the conclusion that the cause of all contagious diseases
must be of a living nature, and this he maintained, although he
had searched in vaecine and small-pox lymph, in the desquamation
of scarlet fever, and in other diseases without success.
Bassi’s discovery and Henle’s doctrine encouraged a number of
investigators, and remarkable results followed. In favus, in herpes
tonsurans, in pityriasis versicolor, fungus threads and spores were
found, and were regarded as being of etiological importance,
inasmuch as the morbid lesions corresponded with the growth of the
particular fungus.
Cholera became especially a subject for research. Swaine,
Brittan, and Budd found micro-organisms in choleraic dejecta.
Davaine described certain monads in the intestinal contents, but no
HISTORICAL INTRODUCTION. 7
causal connection was established between these organisms and the
disease ; and when the cholera disappeared the interest in contagium
vivum waned, and was eclipsed by the question of fermentation.
The discoveries which followed in this subject had a very important
bearing on the micro-parasitic origin of communicable diseases.
Pasteur, following up the researches of Cagniard Latour and
Schwann, demonstrated in 1857 that the lactic, acetic, and butyric
fermentations were produced by micro-organisms.
Previously to this, in 1850, Davaine and Rayer had noted the
existence of little rod-like or filamentous bodies about the size of a
blood corpuscle in the blood of a sheep that had died of splenic fever.
Pollender had seen similar bodies in the blood of cows. Davaine
did not at first pay much heed to this discovery ; but in 1863 he
thoroughly reinvestigated the subject, and conducted a series of
experiments which led him to the conclusion that the actual cause
of splenic fever was an organised heing whose presence and
multiplication in the blood produced changes in that fluid of the
nature of fermentation, resulting in the death of the animal.
These conclusions were not accepted by all, and indeed, the
evidence was so far incomplete that sceptics were justified in con-
sidering that these experiments afforded only a working hypothesis.
But Davaine’s comparison between this disease and fermentation
attracted the attention of Pasteur, whose mind had been fully trained
for entering upon this investigation by the researches which he had
been carrying on in the interval between Davaine’s publications of
1857 and 1863.
Pasteur, as already mentioned, had been working at fermentation,
and his attention was next directed to studying the so-called diseases
of wines, and subsequently to a contagious disease which committed
ravages among silkworms. By laborious researches Pasteur was
able to confirm the belief that this disease of silkworms was due to
the presence of micro-organisms discernible with the aid of the micro-
scope. These oval shining bodies in the moth, worm, and eggs had
been previously observed by Cornalia, and described by Niageli as
Nosema bombycis, and by Lebert as Panhistophyton. But it was
reserved for Pasteur to introduce a means of combating the disease.
Pasteur showed that when a silkworm, whose body contained these
micro-organisms, was pounded up with water in a mortar, and the
mixture painted with « brush on the leaves on which healthy worms
were fed, they would all without fail succumb to the disease.
As the contagious particles were transmitted to the eggs, a
method for preventing the spread of the disease suggested itself.
8 BACTERIOLOGY.
Each female moth was kept separate from the others, and allowed to
deposit her eggs on a small linen cloth. The moth was then pinned
to the corner of the cloth, and left for future examination. When
the time for this arrived, the moth was crushed up with water in a
mortar, and a drop examined under the microscope. When any
trace of corpuscular matter was found to be present, the cloth with
its collection of eggs was burnt; and if not, the eggs were set aside
for use.
Complete as this appears to be as a demonstration of a causal
connection between the micro-organisms and the disease, it could
obviously be objected that there was no distinct proof that the
corpuscular bodies constituted the actual contagium. There was no
isolation of the organisms, no artificial cultivation of them apart
from the diseased moth or worm, and subsequent production of the
disease by means of the isolated organisms. The same objection
was applicable to Davaine’s investigations. Davaine found rods
in association with anthrax, and maintained that they were
causally related ; but others stated that it was possible to inoculate
animals with anthrax blood containing rods, and to produce the
disease without being able to detect the rods again in the blood
of the animal experimented upon. It was also urged that it was
possible to infect with anthrax blood after the rods had disappeared,
and to find a reappearance of the bacilli in the blood of the
inoculated animal.
The well-known fact that anthrax was especially prevalent in
certain seasons and certain localities appeared to lend great support
to these objections. The disease, in fact, was regarded by some
as originating from peculiar conditions of climate and soil. The
fallacies im these objections were, however, rapidly dispelled.
Bollinger, in 1872, pointed out that the blood, from which the rods
had disappeared, was still virulent owing to the presence of the
spores of the bacillus, and that it was owing to the soil being impreg-
nated with these spores that the disease broke out in certain
localities. Yet there still remained many who refused to regard
these particles as living bodies, some looking upon them simply as
crystals; and the question of their importance remained undecided
for several years.
In 1877 Robert Koch published a memoir in which he fully
described the life-history of the anthrax or splenic fever bacillus,
and gave a complete demonstration of the life-history of the micro-
organism, and the definite proofs of its pathogenic properties. He
pointed out how the rods grew in the blood and tissues by lengthen-
HISTORICAL INTRODUCTION. 9
ing and by cross division. Further, that in the blood or in serum
or in aqueous humour they not only grew into long leptothrix
filaments, but they produced enormous numbers of seeds or spores.
He traced, by continuous observation on the warm stage, the whole
life cycle, from the fission of the rods to the formation of spores and
the sprouting of the spores into fresh rods. Further, he carried
on the disease by inoculating from mouse to mouse for several
generations, and observed that in the blood of the animal and in
the swollen spleen the glass-like rods were always to be found.
Pasteur also studied the microbe of splenic fever, and amply
confirmed and extended the observations of Koch by his researches
on the attenuation of the anthrax virus.
Pasteur also met with adverse criticism. Paul Bert argued
that the bacilli were of no importance, because he could destroy
them by exposing material containing them to great pressure, and
yet the material produced the disease on inoculation. But such
measures did not destroy the spores; and finally, Paul Bert was
convinced of his error when Pasteur demonstrated cultures of the
anthrax bacillus in urine, from which successive generations were
started, and that with such cultivations the disease could always be
produced.
It was, however, principally the researches of Koch which
placed the doctrine of contagiwm vivum on a scientific basis.
Koch’s improvements in the methods of cultivation, his recom-
mendation of the necessary microscopical apparatus, his histological
methods for examining these minute organisms, and his famous
postulates for proving beyond controversy the existence of specific
pathogenic micro-organisms, elevated the theory of contagium vivum
to a demonstrated and established fact. The chain of evidence
regarded by Koch as essential for proving the existence of a
pathogenic organism was as follows :—
1. The micro-organism must be found in the blood, lymph, or
diseased tissue of man or animal suffering from or dead of the
disease.
2. The micro-organisms must be isolated from the blood, lymph,
or tissues, and cultivated in suitable media—i.e., outside the animal
body. These pure cultivations must be carried on through successive
generations of the organism.
3. A pure cultivation thus obtained must, when introduced into
the body of a healthy animal, produce the disease in question.
4, In the inoculated animal the same micro-organism must
again ke found.
10 BACTERIOLOGY.
The chain of evidence is still more complete if we can from
artificial cultures obtain a chemical substance which is capable
of producing the disease independently of living micro-organisms.
It is of very little value merely to detect or artificially to
cultivate a bacterium associated with disease. We must endeavour
to establish the exact relationship of the bacteria to disease processes,
and the determination of the true pathogenic microbe is beset
with fallacies. In many diseases bacteria have been regarded as
the actual contagia, until a searching inquiry by other investigators
has shown that the evidence was most unsatisfactory or entirely
misleading. For example, in diseases with lesions of the external
or internal linings of the body, extraneous micro-organisms may
get into the circulation and be swept into the internal organs,
where they either perish in the battle with the healthy tissues
which are opposed to their existence, or they may gain the upper
hand, and set up destructive processes. Such organisms, when
found in association with these diseases, may be discovered in the
blood and internal organs; and though only accidental epiphytes,
often associated with septic complication, they may too readily be
accepted by the enthusiast as the actual contagium of the disease
in question.
It is only when such fallacies are exposed that we are brought
once more face to face with the fact that the nature of the contagium
in hydrophobia, variola, vaccinia, scarlet fever, measles, and many
other diseases, is still undetermined.
CHAPTER II.
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA.
Bacrerta may be considered as minute vegetable cells destitute of
nuclei. They are distinguished from animal cells by being able to
derive their nitrogen from ammonia compounds, and they differ
from the higher vegetable cells in being unable to split up carbonic
acid into its elements, owing to the absence of chlorophyll. Von
Engelmann and Van Tieghem include among the bacteria certain
organisms, named by them Bacterium chlorinum, Bacterium viride,
and Bacillus virens, which are coloured green by this substance, but
it is quite possible that they may be Algz, and further researches
are required before any conclusions are definitely arrived at as to
the exact place these particular organisms occupy in the vegetable
kingdom.
Composition.—For our knowledge of the chemical composition
of bacteria we are chiefly indebted to Nencki. Their constituents
are found on analysis to vary slightly, according to whether the
bacteria are in zoolgcea or in the active state. In the latter condition
they are said to consist of 83°42 per cent. of water. In one hundred
parts of the dried constituents there are the following :—
A nitrogenous body . : ; : : 84:20
Fat. ‘ ' ‘ ; : ; 6:04
Ash . : ; : 3 ; ; : 4:72
Undetermined substances . : ‘ : 5-04
This nitrogenous body is called myco-protein, and consists of
Carbon ‘ : : : ‘ ; 52°32
Hydrogen . . : : : , ‘ 755
Nitrogen . , ‘ ‘ , : 5 14:75
but no sulphur or phosphorus.
The nitrogenous body appears to vary in different species, for
in Bacillus anthracis a substance has been obtained which does not
11°
12 BACTERIOLOGY.
give the reactions of myco-protein, and, therefore, is distinguished
as anthrax-protein.
Considering bacteria as cells, we may speak of the cell-wall and
the cell-contents. The cell-wall consists of cellulose, or, according
to Nencki, in the putrefactive bacteria of myco-protein. It may be
demonstrated by the action of iodine, which contracts the proto-
plasmic contents, and renders the cell-wall visible. By staining
cover-glass preparations of the anthrax bacillus by the method of
Gram, the rods are at first uniformly stained, by subjecting them to
iodine solution the protoplasmic contents are contracted, and alcohol
decolorises the sheath, which may be then stained in contrast, with
eosin.
The cell-wall may be either pliable or rigid. liability is
observed in the long filaments, which are endowed with a slow
vermicular movement, while rigidity accounts for the maintenance
of the characteristic form of several species, such as spirilla.
The cell-protoplasm yields myco-protein. In some itis homogene-
ous, and in others granular. The action of the aniline dyes indicates
a close relation to nuclear protoplasm, though all nuclear stains are
not suitable for bacteria. In some cases also the bacteria remain
stained under the influence of a reagent, which removes the colour
from nuclei. The power of fixing the stain is not always present,
and indicates a difference in the protoplasm of different species.
Thus in staining phthisical sputum, the nitric acid removes the
stain from all bacteria and bacilli present, with the exception of the
tubercle bacillus. This difference in the protoplasm of different
species is also illustrated by the necessity, in many cases, of using
special processes, owing to the ordinary methods being unsatisfactory
or not producing any result.
The protoplasm of some bacteria contains starch granules; thus
Clostridium butyricum gives the starch reaction with iodine.
Sulphur granules are present in some species of Beggiatoa which
thrive in sulphur springs. The colouring-matter of the pigmented
bacteria is probably external to the cell asa rule: for example, in
Micrococcus prodigiosus the pigment granules are distictly between
the cells; on the gther hand, in Beggiatoa roseo-persicina, or the
peach-coloured bacterium, the special pigment bacterio-purpurin
appears to be dissolved in the cell protoplasm. In Bacillus
pyocyaneus the pigment is certainly not localised entirely in the
cell, for it becomes rapidly diffused in the surrounding medium,
considerably beyond the confines of the growth itself.
In several species, either as a result of a secretion from the cell or
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 13
of the absorption of moisture and consequent swelling of the outer
layer of the cell-wall, a mucinous or gelatinous envelope develops
around them. This envelope may form a capsule, such as we meet
with in certain bacteria found in the rusty sputum of pneumonia, and
in Micrococcus tetragenus ; or it may occur as a continuous sheath
around a chain of bacteria, which by its disappearance sets free
the individual links.’ The capsule is soluble in water, and under
some circumstances is difficult to demonstrate. In the preumo-
coccus of Friedlinder the capsule disappears on cultivation, but
reappears in preparations made from an inoculated animal. In the
pleuritic fluid of a mouse these cocci are often found with a parti-
cularly well-marked capsule, and in other encapsuled cocci the extent
of the envelope has been observed to vary considerably in the same
species of bacterium.
When this gelatinous material forms a matrix, in which
numbers of bacteria are congregated in an irregular mass, we have
what is termed a zo00glea. The zooglean stage is a resting stage,
often preceded or followed by a motile stage. Thus bacteria may
be present in a solution in an active state, and after a time a scum
or pellicle forms on the surface of the liquid, which consists of
zooglea. At the edges of the zooglea, individuals may be seen to
again become motile, and after detaching themselves to swim off in
the surrounding fluid.
The zooglean stage may be observed sometimes in cultivations in
broth, and also in nutrient gelatine which has become liquefied.
The inoculated bacteria grow and multiply, and after a time a film
appears on the surface of the liquefied layer. In cultivations on
potato the appearances in this stage are varied, and sometimes
extremely characteristic. In the case of a bacillus which readily
develops on unsterilised potatoes, the zooglea may spread over the
cut surface, forming a pellicle which can be raised en masse like
a delicate veil. Another bacillus forms a zooglea, consisting of a
tenacious layer which can be drawn out in long stringy threads.
In Ascococcus Billrothii the gelatinous envelope develops to such
an enormous extent that it forms the characteristic feature of the
species. (Fig. 1.)
Form.—The individual cells vary in form, and may either
remain isolated or attached to each other. Round cells and egg-
shaped cells are called cocci, The spherical form is the most
common, but cocci are occasionally exclusively ovoid, as in Strepto-
coceus bombycis. The giant cocci of some species are spoken of as
megacocci, to distinguish them from the ordinary cocci, or micrococci.
14 BACTERIOLOGY.
The fission by which the cocci increase may take place in one
direction only, and if the two resulting cells remain attached to
each other they form a diplococcus. If these two cells again divide,
and the resulting cells remain linked together, we get a chain or
rosary, termed streptococcus. These chains may consist of a few
individuals linked together, or of several hundreds, in which case
the chains are generally curved or twisted. When the division
occurs in two directions, so that four cocci result, a tetrad or
merismopedia is formed ; when in three directions, one coccus divides
into eight, and the result is a packet form or sarcinacoccus.
Immediately after division, the daughter cells are not perfectly
circular, but are flattened or facetted where they are opposite to
Fic. 1.—Ascococcus Bintrorum, x 65. [After Cohn.]
each other. They gradually become rounded off, and each daughter
cell is then ready to divide in its turn. In other cases the cocci
after division only form irregular heaps or collections like bunches
of grapes. This form is sometimes distinguished as staphylococcus,
but it cannot be considered an important feature. When we find
irregular masses of cocci united by intercellular substance and
embedded in a tough gelatinous matrix, the form is described as
ascococcus.
Another type is the rod, characteristic of bacterium and bacillus.
The rods may vary considerably in length. The very short rods
with rounded ends are difficult to distinguish from the oval cocci,
but ‘differ in that a rod, however short it may be, must have
two sides parallel. The vibrio or bent rod may be considered as
the connecting link between the rods and the corkscrew forms or
DESCRIPTION OF PLATE I.
Bacteria, Schizomycetes, or Fission Fungi.
1, Cocci singly and varying in size. 2. Cocci in chains or rosaries (strepto-
coceus), 3. Cocci in a mass (staphylococcus). 4 and 5. Cocci in pairs
(diplococcus). 6. Cocci in groups of four (merismopedia). 7. Cocci in packets
(sarcina). 8. Bacterium termo. 9. Bacterium termo x 4000 (Dallinger and
Drysdale). 10. Bacterium septicemie hemorrhagice. 11. Bacterium pneu-
monie croupose. 12. Bacillus subtilis. 13. Bacillus murisepticus. 14.
Bacillus diphtheria. 15. Bacillus typhosus (Eberth). 16. Spiriliwm wndula
(Cohn). 17. Spirillum volutans (Cohn). 18. Spirillwm cholere Asiatice.
19. Spirillum Obermeiert (Koch). 20. Spirocheta plicatilis (Fliigge). 21.
Vibrio rugula (Prazmowski). 22. Cladothrix Férsteri (Cohn). 23. Cladothria
dichotoma (Cohn). 24. Monas Okenii (Cohn). 25. Monas Warmingii (Cohn).
26. Rhabdomonas rosea (Cohn). 27. Spore-formation (Bacillus alvei). 28.
Spore-formation (Bacillus anthracis). 29. Spore-formation in bacilli cultivated
from a rotten melon (Frankel and Pfeiffer). 30. Spore-formation in bacilli
cultivated from earth (Frankel and Pfeiffer). 31. Involution-form of Crenothria
(Zopf). 32. Involution-forms of Vibrio serpens (Warming). 33. Involution-
forms of Vibrio rugula (Warming). 34, Involution-forms of Clostridiwm
polymyxa (after Prazmowski). 35. Involution-forms of Spirillum cholere
Asiatice. 36. Involution-forms of Bacterium aceti (Zopf and Hansen).
37. Spirulina-form of Beggiatoa alba (Zopf). 38. Various thread-forms, of
Bacterium merismopedioides (Zopf). 39. False-branching of Cladothria (Zopf).
Fia. 30
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BACTERLA, SC HIZOVYCETES, OR: FISSION FUNGI,
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 15
spirilla, Lastly, we have the filamentous forms, which may be
straight, leptothria, or wavy, spirocheta, or the wavy thread may
be looped and entwined on itself, spirulina.
The term involution form is applied to certain peculiar shapes,
which result more especially in bacteria grown under abnormal
conditions. They are round, oval, pear-shaped, or club-formed
enlargements.
Movement.—Many bacteria are devoid of movement through-
out the whole of their life history. Others, during certain stages of
their life cycle, and possibly some forms always, are endowed with
locomotive power. The character of the movement is very varied,
and ranges from a slow undulatory motion to one of extreme
rapidity. Many appear to progress in a definite direction. Others
move continuously, first in one direction and then in another, and
others again seem to hesitate before altering their course. They
may either glide along smoothly or progress with a tremulous motion.
Fic. 2.—SprrocH£TA FROM SEWAGE WaTER, x 1200.
They appear to be able to avoid obstacles, and to set themselves
free from objects with which they have accidentally come into
contact. Vibrios have a peculiar serpentine movement, but other
forms, such as the commonly known Bacterium termo and segments
of spirilla, such as comma-bacilli, revolve around their long axis
as well as make distinct progression. The complete spirilla are
characterised by the familiar corkscrew movement. With regard
to cocci there is some doubt as to whether they are endowed with
independent movement, any quivering or oscillation being generally
regarded as only Brownian or molecular. In some straight thread-
forms, which are motile, the movement is very slow and vermicular
in character, but in wavy threads, such as the Spirocheta plicatilis,
there is not only an undulatory motion, with rapid progression across
the field of the microscope, but if they are confined by more or
less débris, they give very peculiar and characteristic spasmodic.
movements. (Fig. 2.)
The rod-forms of Proteus vulgaris exhibit very extraordinary
16 BACTERIOLOGY.
movements on the surface of solid nutrient gelatine. Groups of
rods may be observed to pass each other in opposite directions,
Single individuals meet and progress side by side, or one or more
individuals may part from a group and glide away independently,
Occasionally a number of rods progress in single file. It is, however,
difficult to believe that these movements can occur on a solid surface.
Fig. 3.—FLAGELLA.
1. Coccus with flagellum. 2. Similar coccus dividing, with two flagella. 3. Colony
of flagellated macrococci of Beggiatoa rosco-persicina. 4, Short rod from the
same Beggiatoa with flagella [all after Zopf]. 5. Bacillus with flagella [from
a photograph by Koch]. 6. Bacillus subtilis |after Brefeld]. 7, 8. Short rod-
forms of Beygiatoa roseo-persicina with one flagellum [after Zopf]. 9. Very
long rod of the same, with flagellum at both ends [after Warming]. 10. Vibrio,
with double flagellum at each end [after Warming]. 11. Vibrio, with flagella
[from a photograph by the author]. 12. Spirillum with flagella [from a photo-
graph by Koch]. 13. Spirillum with flagella [after Zopf]. 14. Spirillum with
double flagella [after Zopf]. 15. Beygiatow roseo-persicina, with a triple
flagellum at one end; and 16, with a double flagellum at both ends [after
Warming].
The author is inclined to believe that there is an almost inappreciable
layer of liquid on the surface of the gelatine, which is expressed
after the gelatine sets. In tubes of nutrient agar-agar gelatinised
obliquely and then kept upright the liquid so expressed collects at
the bottom of the sloping surface.
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 17
The means by which bacteria are endowed with the power of
spontaneous movement and of progression may still be said, in
some cases, to be unsettled. The author has watched the move-
ment of long slender threads in sewage-contaminated water, which
could only be explained by the inherent contractility of the proto-
plasmic contents; for if any drawing or propelling organ existed
in proportion to the length of the organism, it would probably have
been visible. But in many cases the organism is provided with a
vibratile lash or flagellum at one end, or with one or more at both
ends, or with numerous lateral and terminal flagella.
Some observers believe that the movement of cocci is due to the
Fic. 4.—BacILLus MEGATHERIUM.
a. A chain of rods, x 250. The rest x 600.
b. Two active rods.
d to f. Successive stages of spate. -formation.
h tom. Successive stages of germination.
[After De Bary.]
existence of a flagellum. In Bacterium termo the existence of a
lash at either end was first determined by the researches of Dallinger
and Drysdale. In motile bacilli, such as the hay bacillus and
Bacillus ulna, and in vibrios and spirilla, the flagella can be readily
recognised by expert microscopists with the employment of the best
lenses, and, what is of equal importance, proper illumination. They
are objects of extreme delicacy and tenuity, and in stained prepara-
tions may be absent from retraction or injury. Koch succeeded
in photographing them after staining with logwood, which turned
them a brown colour. The author has observed them in vibrios in
preparations stained with gentian violet, from which also they have
been photographed, in spite of the violet colour, by the use of
2
18 BACTERIOLOGY,
isochromatic dry plates, and more recently special methods have
been introduced, by Léffler and others, by which they can be stained
and photographed with comparative facility.
It is not certain whether the flagella are extensions of the cell-
wall, or derived from the internal protoplasm. Van Tieghem holds
the first view, and does not regard them as motile organs at all.
Zopf, on the other hand, adheres to the second view, aud moreover
believes that they can be retracted within the cell-wall.
Reproduction.—Bacteria multiply by fission and by processes
which may be considered as representing fructification. The
Fic. 5.—CLosTRIDIUM BUTYRICUM, x 1020.
B. Stages of spore-formation.
C. Stages of germination.
‘[After Prazmowski.]
bacteria exhibiting the latter processes have been divided into two
groups, distinguished by the formation of endospores in the one and
of arthrospores in the other. In the process of fission the cell first
increases in size, and a transverse septum forms from the cell-wall,
dividing the internal protoplasm into two equal parts; these may
separate and lead an independent existence, or remain linked
together. In chains of cocci the individual cells are easily visible
and distinct, but in the thread-forms resulting from the linking
together of rods, as in the anthrax bacillus, the composition of the
thread is only demonstrated by the action of reagents.
Endospore formation may be conveniently studied in Bacillus
anthracis, Bacillus megatherium, or Bacillus subtilis. The proto-
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 19
plasm becomes granular, and at certain points in the thread a speck
appears, which gradually enlarges and develops into a circular
or egg-shaped, sharply defined, highly refractive body. The spore
grows at the expense of the protoplasm of the cell, which in time,
together with the cell-wall, entirely disappears, and the spore is set
free. These phenomena are best seen in an immotile bacillus in a
drop-cultivation on a warm stage; the whole process may then be
observed continuously from beginning to end. Spores may form in
each link of the thread, so that a regular row results, or they may
occur at irregular intervals. Spore-formation also occurs in bacilli
which do not develop into leptothrix filaments. The spores may
develop in the centre or at one end of the rod. In the tetanus
bacillus a spore develops at the extreme end, producing the appear-
ance of a drum-stick. The spore may be considerably wider, but
is never longer than the parent cell.
Fic. 6.—LEUcONOSTOC MESENTEROIDES ; COCCI-CHAINS WITH ARTHROSPORES
(after Van Tieghem and Cienkowski).
Arthrospore formation is illustrated in Leuconostoc mesenteroides.
Certain elements in the chain of cocci, apparently not differing from
the rest, become larger, with tougher walls, and more refractive.
The remaining cells die, and these cells having acquired the pro-
perties of spores are set free, and can reproduce a new growth in
any fresh nourishing soil. That this occurs in all species which
do not form endospores is at present only a supposition.
Spores are invested by a thick membrane, which is believed to
consist of two layers. To this they probably owe the property they
possess of retaining vitality when desiccated, and of offering a
greater resistance to the action of chemical reagents and heat than
the parent cells. \
Spore-formation has been regarded by some as occurring when
the nourishing soil is exhausted, thus providing for the perpetuation
’
20 BACTERIOLOGY.
of the species. In anthrax the bacilli do not form spores in the
living body, but when the animal dies the development of spores
takes place, and hence the danger of contaminating the soil if the
body is disposed of by burial. Klein, however, has pointed out that
if mice and guinea-pigs which have died of anthrax are kept un-
opened, the bacilli simply degenerate and ultimately disappear.
Thus there is good reason for believing that spore-formation is
not due to exhaustion of the pabulum, but probably free access to
oxygen constitutes an important factor in inducing this condition.
If we inoculate a potato with anthrax, copious spore-formation
oceurs, though we cannot consider that the nourishing soil has been
exhausted. But we have in this case the surface of the potato
freely exposed to the air in the damp chamber. In the same way,
in cultivations on agar-agar solidified obliquely (so as to get a large
surface), spore-formation readily takes place. Contamination of
Fic. 7.—SPoRE-BEARING THREADS oF Bacittus ANTHRACIS, DouBLE- SEINE:
WITH FUCHSINE AND METHYLENE BLUE, x 1200.
the ground results, therefore, from animals in which a ae
examination has been made and the blood and organs freely exposed
to the air; or from carcasses the hides of which have been soiled
with excretions, and with blood which issues from the mouth and
nostrils before death.
When spores are introducéd into a suitable medium at a favour-
able temperature they develop again into rods. The spore loses its.
sharp contour, and, at one pole or on one side, a pale précess bursts
through the membrane, gradually growing into a rod from which
the empty capsule is thrown off.
Spores differ from the parent cells in their behaviour to staining
reagents. Like them, they can be stained with aniline dyes, but
not by the ordinary processes. They require to be specially treated.
This is probably due to the tough capsule, which must be altered
or softened by heat or strong acid, until it allows the stain to
penetrate.
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 21
Once stained, they again differ from the parent cells in resisting
decolorisation ; this fact is taken advantage of to double-stain spore-
bearing bacilli (Fig. 7).
In staining micro-organisms, the protoplasm is sometimes broken
up into irregular segments or granules, as in many spirilla, and we
may add the bacilli of tuberculosis and leprosy. The beaded
appearance of the tubercle bacillus is well known. Some observers
have regarded the beads, others the bright spaces -between them,
as spores. But spores in unstained preparations appear as glistening
bodies with sharp contour. They do not stain at all, or very little,
by the ordinary processes. These considerations led the author to
stain and examine tubercular sputum and pure-cultures under
careful illumination, and with such lenses as Powell and Lealand’s
gs in.hom,imm. The tubercle bacillus in sputum (Fig. 8), as a rule,
!
Fic. 8.—BactLur or TUBERCLE In SPuTuM, x 2500 (from photographs).
consists of a very delicate sheath, holding together a number of
deeply stained granules, for the most part round or cylindrical, with
irregular contour, and differing considerably in size, while the light
interspaces are seen to vary in form according to the shape of the
granules, On the other hand, particularly in old cultures, more or
less spherical, sharply defined bodies are observed in the bacilli, and
also set free. These are the true spores of the tubercle bacillus, ~
and are quite distinct from the irregular granules. There can be
no doubt that a tubercle bacillus consists of a very delicate sheath,
with protoplasmic contents which have a great tendency to break
up or coagulate into little segments or roundish granules, partly
owing to their age and the conditions under which they are grown,
and partly to the treatment they are subjected to in making a
microscopical preparation. This does not always occur, for the
bacilli at times are not beaded, but are stained in their entirety.
Tn the leprosy bacilli a similar appearance occurs. In stained
22 BACTERIOLOGY.
sections the rods have a beaded appearance, but the intervals between
the granules are sometimes very long, and occasionally the protoplasm
appears to have collected only at the extreme ends of the rod.
The appearances in the case of the bacillus of glanders and the
bacillus of hemorrhagic septicemia may be similarly explained.
The fact that tubercular sputum preserves its virulence for
several months, even after desiccation, is to be attributed to the
formation of spores. Babés claims to have succeeded in differen-
tiating them by double staining.
In his definition of spirilla, Zopf gives the spore-formation as
absent or unknown. In comma-bacilli in sewage water the author
has often noted appearances suggestive of refractive spores; and
the same also may be observed in vibrios, differing by their regular
contour from the irregular spaces occasionally observed in stained
preparations; but they are only vacuoles.
+ gd
a 2,
es
Fic. 9.—ComMa-BAcILLI In SEw- Fic. 10. Visrios In WATER CON-
AGE WATER, STAINED WITH TAMINATED WITH SEWAGE,
GENTIAN VIOLET, x 1200. x 1200.
Respiration and Nutrition.—Like all a-chlorophyllous vegeta- -
bles, bacteria require for their nutrition oxygen, nitrogen, carbon,
water, and certain mineral salts. Many require free access to oxygen,
others can derive it from the oxidised compounds in the medium
in which they grow. Pasteur divided bacteria into two great classes
—the aerobic and anaerobic, and considered that the latter not
only had no need of oxygen, but that its presence was actually
deleterious. Though this view must be considerably modified, the
terms are convenient, and are still retained. They are well illus-
trated by the bacillus of anthrax, and the bacillus of malignant
edema; anda simple plan of demonstration has been employed
by the author. A fragment of tissue from the spleen, for example,
known to contain anthrax bacilli, is deposited with a sterilised
inoculating needle, with the necessary precautions, on the surface of
nutrient agar-agar in a test-tube ; another tube of nutrient agar-
agar is liquefied, and when cooled down almost to the point of
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 23
gelatinisation, a part is poured into the first tube, so that when it
sets the piece of tissue is completely embedded. A piece of tissue
from an animal suffering from malignant cedema is treated in the
same way, and the tubes are placed in the incubator. If we
examine them after two or three days, we shall find no change in
the anthrax tube; the bacillus being eminently aerobic, no growth
whatever has occurred. In the tube containing the bacilli of
malignant cedema there will be a more or less characteristic
cultivation.
The nitrogen which is essential for building up their protoplasm
can be obtained either from albumins, or from ammonia and its
derivatives. That the albumins can be dispensed with was shown
by Pasteur, who employed an artificial nourishing solution consti-
tuted upon a formula representing the essential food constituents.
Carbon is derived from such substances as cane sugar, milk
sugar, and glycerine, and, in some cases, by the splitting up of
complex proteid bodies.
Water is essential for their growth, but deprivation of water
does not kill all bacteria. Desiccation on potato is employed for
preserving some micro-organisms, as a new growth can be started,
when required, by transferring some of the dried potato to fresh
nourishing ground. Comma-bacilli, on the other hand, are
destroyed by drying. Sugar is used in making preserves, because
by abstracting water it prevents the development of micro-
organisms.
Mineral or inorganic substances, such as compounds of sodium
and potassium, and different phosphates and sulphates, are necessary
in small proportions.
CIRCUMSTANCES AFFECTING THE GROWTH OF BacTERIA.
Nature of the Soil—_Though we know the elements necessary,
we are, nevertheless, as yet unable to provide a pabulum suitable for
all kinds of bacteria. Thus we are quite unable to cultivate some
species artificially. Others will only grow upon special media.
Many grow upon nutrient gelatine; but some species only if it be
acid or alkaline respectively. Whether in the latter case this is due
purely to the reaction or to the presence of the particular ingredients
is an unsettled point. Though the comma-bacillus of Koch, like the
majority of organisms, grows best on an alkaline medium, yet it
is well known to flourish at the temperature of the blood on the
surface of potato, which is acid.
24 BACTERIOLOGY.
Temperature.—The influence of temperature on bacteria will be
found to vary according to the species, but still for the majority we
may distinguish a maximum, optimum, and minimum temperature.
Many grow best at the temperature of the blood, and hence the
value of nutrient agar-agar, which is not liquefied at 37° C. The
tubercle bacillus will only grow satisfactorily at a temperature
varying between 30° C. and 41°C. On the other hand, many forms
grow between the limits of 5° C. and 45°C. At these temperatures
their functional activity is paralysed, but they are not destroyed,
for by removal to favourable conditions they spring again into life.
Bacteria seem to have a special power of resisting the effects of cold.
It has been stated that comma-bacilli exposed to a temperature
of—10° C. for an hour, and bacilli of anthrax after exposure to a
temperature of —110° C., still retained their vitality. Temperatures
over 50° to 60° C. destroy most bacteria, but not their spores; spores
of anthrax retain their vitality after immersion in boiling water, but
are destroyed by prolonged boiling. Roughly speaking, all patho-
genic bacteria grow best at the temperature of the blood, and
non-pathogenic bacteria at the ordinary temperature of the room.
Movement.—Bacteria probably grow best when left undisturbed.
Violent agitation of a vessel in which they are growing certainly
retards their growth, but a steady movement is stated not to affect
it; at any rate, anthrax bacilli grow with enormous rapidity in the
blood-vessels, in spite of the circulation.
Compressed Air.—Paul Bert maintained that a pressure of
twenty-three to twenty-four atmospheres stopped all development
of putrefactive bacteria. Oxygen, under a pressure of five or six
atmospheres, is stated to stop their growth. Other observers have,
however, obtained different results,
Gases.—Hydrogen and carbonic acid are stated to stop the
movements of the motile bacteria. Chloroform is believed to arrest
the changes brought about by the zymogenic species.
Electricity—Cohn and Mendelssohn found that a constant
galvanic current produced a deleterious effect owing to electrolysis.
At the positive pole the liquid became distinctly acid, and at the
negative pole distinctly alkaline. With a weak current there
appeared to be no effect, two powerful cells at the very least being
necessary.
Light.—Downes has shown that sunlight is fatal to putrefactive
bacteria. This is believed to be due to a process of induced hyper-
oxidation, from which living organisms ordinarily are shielded by
protective developments of the cell-wall, or of colouring-matter, —
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 25
which cut off injurious rays. Duclaux has investigated the same
subject, and observed that micrococci were more sensitive to sun-
light than the spore-bearing bacilli. Engelmann has described a
bacterium whose movements cease in the dark, and Zopf states that
in his cultures of Beggiatoa roseo-persicina the growth was much
more strongly developed on the side of the vessel facing the light.
Arloing, Marshall Ward, and Dieudonné have studied the effect
of the sun’s rays on anthrax spores, and on chromogenic and
other bacteria, and maintain that they are bactericidal. The
effect is due chiefly, if not entirely, to the blue rays.-
Chemical Reagents.—Many substances, such as carbolic acid,
corrosive sublimate, chlorine, bromine, have a marked effect upon
the growth of bacteria. This will be more fully described in
another chapter. In several cases the bacteria themselves secrete
a substance which is injurious to their future development.
: Propucts oF GROWTH.
Bacteria may be grouped together according to the changes pro-
duced in the media in which they grow. Thus we have pigment-
forming, phosphorescent, fermentative, putrefactive, nitrifying, and
disease-producing bacteria.
Chromogenic or pigment-forming bacteria elaborate during their
growth definite colour stuffs. Such species are exemplified by Bacillus
violaceus, which produces a striking purple growth; Bacillus
pyocyaneus, which secretes pyocyanin, a substance which has been
isolated and obtained in a crystalline form ; Micrococcus prodigiosus,
which produces a pigment allied to fuchsine; Beggiatoa roseo-per-
sicina, which is characterised by the presence of bacterio-purpurin ;
Sarcina lutea, Bacillus cyanogenus, and many others.
Photogenic, or light-producing, bacteria are found more especially
in sea-water. There are several species of phosphorescent bacilli,
and according to Beyrinck the best medium for their cultivation is
fish-broth made with sea-water. Photographs can be obtained of
cultures by their own light.
Zymogenic or ferment bacteria produce their changes in non-
nitrogenised media. Bacterium aceti, by its growth produces the
acetic fermentation in wine by which alcohol taking up atmospheric
oxygen is converted into vinegar :—
C7H5O + 0? = C’H'0? + H70.
The fermentation of urine, by which urea is converted into carbonate
of ammonia, can be brought about by several micro-organisms, but
26 BACTERIOLOGY.
notably by the Hated ure, The change produced is represented
by the following formula :—
°0 = (NH) 200°,
co(NE + 2120 = (NH)
Clostridium butyricum converts the salts of lactic acid into
butyric acid, producing the butyric fermentation in solutions of
starch, dextrine, and sugar. These bacteria are agents in the
ripening of cheese, and the production of sauerkraut.. Thus, in a
solution neutralised with calcium carbonate :—
2[Ca(C*H50")'] + H20 = Ca(C!H70%)? + CaCO* + 300? + He.
In the so-called viscous fermentation of wines, Streptococcus viscosus
produces a gummy substance. According to Pasteur, the change
may be thus represented :—
25(C"H™0") + 25(H20) = 12(C"H”0") + 24(C°HO%) +
12(CO2) + 12(H20).
And as another example, the Bacillus acidi lactici may be mentioned,
through the agency of which sugar of milk is converted into lactic
acid :—
CR AAO = 4(C%H'0*).
Saprogenic or putrefactive bacteria play a most important part
in the economy of nature. They produce changes allied to fermenta-
tion in complex organic substances, Their action on proteids,
according to Hoppe-Seyler, may be compared to digestion ; bodies
like peptones are first produced, then leucin, tyrosin, and fatty
acids ; lastly indol, phenol, sulphuretted hydrogen, ammonia, carbonic
acid, and water. They abstract the elements they require, and the
remainder enter into new combinations. Associated with the forma-
tion of these substances are certain bodies which have a poisonous
effect when introduced into animals. These poisonous alkaloids,
ptomaines, produce a septic poisoning, which must be distinguished
from septic infection. The effects of septic poisoning depend on the
dose, whereas the effects of septic infection are, to a certain extent,
independent of the dose. A small quantity of a septic poison may
produce only transient effects, and a relatively large quantity may
be necessary to produce vomiting, rigors, and death. Septic in-
fection, on the other hand, may result equally from a small dose,
because the poison introduced is a living organism which is capable
of propagation and multiplication. Our knowledge of these
alkaloids is largely attributable to the researches of Selmi, Gautier,
and Brieger, and the result of their work will be referred to again,
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 27
Nitrifying bacteria play a very important part by providing
plant life with a most necessary food. They occur in the soil, and
two kinds have been described—the one kind converting ammonia
into nitrous acid, and the other changing nitrous into nitric acid.
To Winogradsky and Frankland we are principally indebted for our
knowledge of these bacteria.
Pathogenic bacteria are those which are genetically related to
disease. Many organisms have been supposed to be pathogenic, or
have been described in connection with diseases, which are only
saprophytic associates. By saprophytic we mean organisms which
feed upon dead organic matter. They include many forms which
are found on the skin, in the intestinal canal, and sometimes in the
internal organs, especially the liver and kidneys; the tissues have
lost their vitality, and the organisms, through some lesion, have
been carried into the circulation.
That many organisms are causally related to disease, there is
strong evidence in proof. No organism can be considered to be pro-
ductive of disease unless it fulfils the conditions which have been
laid down by Koch. Great stress must be laid upon the importance
of successive cultivation through many generations, as the objection
that a chemical virus may be carried over from the original source
is thus overcome. Any hypothetical chemical poison carried over
from one tube to another would, after a great number of such
cultivations, be diluted to such an extent as to be inappreciable
and absolutely inert.
Though we may accept as a fact the existence of pathogenic
organisms, we are not in all cases in a position to assert the means
by which they produce their deleterious or fatal effects, Many
theories have been propounded. It has been suggested that the
pathogenic organisms may be compared to an invading army.
The cells or phagocytes arrayed against them endeavour to as-
similate and destroy them, but perish themselves in the attempt.
This might explain the breaking down of tissue, and the for-
mation of local lesions, but does not assist us in understanding
the fatal result in thirty-six to forty-eight hours produced by the
inoculation of the bacilli of anthrax. Another view is that the
invading army seizes upon the commissariat, appropriating the
general pabulum, which is so essential to the life of the tissues.
This would hardly account for so acute and fatal a result as in
anthrax, but would lead one to expect symptoms of inanition and
gradual exhaustion. Moreover, against this theory we have the
fact that death may result, in some cases, with the presence
28 BACTERIOLOGY.
of comparatively few bacilli in the blood; and, again, the
blood may teem with parasites such as the flagellated monads in
well-nourished, healthy-looking rats, without, apparently causing
any symptoms whatever. In the same category may be placed the
theory that eminently aerobic organisms seize upon the oxygen of
the blood and produce death by asphyxia. Another explanation is
afforded by the suggestion of interference with the functions of the
lung and kidney by mechanical blocking of the capillaries. Here
the same objection is met with in the case of anthrax, the same
fatal result may occur with only a few bacilli, while other cases
yield very beautiful sections, looking like injected preparations from
the mapping out of the capillaries with the countless crowds of
bacilli.
Putrefactive bacteria derive their necessary elements from com-
plex organic substances, and accompanying the residue we find the
presence of poisonous substances. Pathogenic bacteria, in a similar
way, give rise to virulent poisons. Anthrax bacilli produce poisonous
principles in the blood which cause death, independently of the
number of bacilli, provided there are sufficient present to develop a
fatal dose.
It has been also suggested that possibly a special ferment is
secreted by some organisms, and that by the changes ultimately
wrought by the action of this ferment the symptoms and phe-
nomena of disease arise. We have an analogy with this theory
in the alkaline fermentation of urine by means of the torula urex.
By the researches of Musculus, and later of Sheridan Lea, it has
been shown that a ferment is secreted by the cells which can be
isolated in aqueous solution, and is capable of rapidly inducing an
active fermentation of urea.
We can now understand how it is that in anthrax or in tuber-
culosis we may find the presence of only a few bacilli, or that in
tetanus we can have such a violent disturbance of the system
produced by the presence of very few micro-organisms. We may
conceive that different species of bacilli may vary greatly in their
power of producing a toxin or secreting a-ferment, just as the
elaboration of pigment is much more marked in some species than
in others; thus it need not follow that the number of micro-
organisms bears any relation to the virulence or activity of the
substance they produce. There is, however, yet another factor in
the production of disease. We know that in health we are proof
against most of these micro-organisms ; if it were not so, we should
all rapidly fall victims to the tubercle bacillus or others, which in
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 29
health we inhale with impunity. We know that a microbe may only
cause a local lesion in one animal, but death in another. It is still
more striking that the same micro-organism, as is the case with
anthrax, may have no effect whatever upon certain species of
animals, though it is deadly to others. Again, an animal naturally
susceptible to the effect of a pathogenic organism may be rendered
proof against it. These matters will be discussed in a future.
chapter.
DistriButTION OF BACTERIA.
Bacteria are commonly described as ubiquitous. They are ever
present in the air, though not in such exaggerated numbers as is
commonly supposed. In nutrient media exposed to the air one is often
astonished at times at the comparatively few bacteria which develop
in comparison to the amount of floating matter, such as mineral
particles, scales, spores of fungi and débris known to be present.
In water they are also present in considerable numbers, though of
course varying according to the character of the water. Wherever
there is putrefaction, they are present in vast numbers. In the soil,
in sewage, in the intestines and, in uncleanly persons especially,
on the skin and between the teeth, various species may always he
found, but in the healthy blood and healthy tissues bacteria are
never present. Ina previous chapter the method of examining the
blood of living persons has been described, and there is, by this
means, ample opportunity for satisfying oneself that bacteria are
never to be found in the blood in health. The organs removed from a
perfectly healthy animal, with the necessary precautions, and placed
in sterilised media, can be kept indefinitely without undergoing
putrefaction, or giving any development of bacteria. This has been
established by many observers, notably Cheyne and Hauser; and
the results of former observers to the contrary must be attributed
to imperfect methods admitting of accidental contamination.
1
CHAPTER III.
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA.
fn the previous chapter several conditions were alluded to which
affected the growth of bacteria, such as the nature of the
nutrient soil, temperature, light, and electricity. The effect of
certain chemical substances, and of excessive heat and cold, was
also mentioned; but this constitutes a subject of such practical
importance that it must be considered more fully. a
Agents which retard the growth of bacteria are generally spoken
of as antiseptics, as distinguished from disinfectants which altogether
destroy their vitality.
Though chemical disinfectants, or germicides, when diluted, act
as efficient antiseptics, the converse, that an antiseptic in a suffi-
ciently concentrated form will act asa disinfectant, is not the case.
The term ‘ antiseptic,” indeed, should be restricted to those sub-
stances or agents which arrest the changes bacteria produce, but
which do not prevent their springing into activity when removed
to favourable conditions. Thus excessive heat, which destroys
bacteria and their spores, is a true disinfectant ; and excessive cold,
which only benumbs them, retarding their development without
killing them, is an antiseptic.
Spores have a greater power of resisting the action of these
various agents than the parent cells, and many species of micro-
organisms differ from each other in their resisting power. Au
exact knowledge of the subject can, therefore, only be based upon
investigations which will determine the effect of these agents upon
pure cultivations of the different micro-organisms causally related to
putrefaction and disease. In the latter case, especially, this is not
possible in the present state of our knowledge. In some cases of
communicable disease there is considerable doubt as to the etiological
importance of the organisms which have been described; in other
cases no organisms have as yet been discovered, or the organisms
30
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA, 31
cannot be artificially cultivated, or the disease is not reproduced by
inoculation, so that there is no means of testing whether the agents
have had any effect. One can, therefore, only draw general
conclusions by selecting some well-known pathogenic and non-
pathogenic micro-organisms, and considering the influence of
chemicals, of hot air and of steam upon them, as representing the
effect upon the various contagia of disease and the causes of
putrefaction.
Such knowledge must necessarily prove of the greatest im-
portance: to the sanitarian, who is concerned in preventing the
spreading of disease and in the disposal of putrefactive matter; to
the surgeon, who is anxious to exclude micro-organisms during
surgical operations, and to arrest the development of bacteria which
have already gained an entrance in wounds; to the physician, in
the treatment of micro-parasitic diseases.
The sanitarian and the surgeon must profit directly by such
experiments, for in the disinfection of clothes and the sick-room by
the one, and in the application of antiseptic dressings and lotions
by the other, the micro-organisms are encountered, as in the experi-
ments, outside the living body.
The physician, on the other hand, is principally concerned in
_ dealing with micro-parasites when circulating in the blood, or
carrying on their destructive processes in the internal tissues. So
far as our knowledge at present goes, the physician can avail him-
self but little of the effect of the direct application of the substances
which have been found to retard or destroy the growth of the
organisms in artificial cultivations, for the concentrated form. in
which they would have to be administered would prove as deleteri-
ous or as fatal to the host as to the parasites. Thus Koch has
stated that to check the growth of the anthrax bacillus in man it
would be necegsary that there should be twelve grammes of iodine
constantly in circulation, and that the dose of quinine necessary
to destroy the spirilla of relapsing fever would be from twelve to
sixteen grammes. The retarding influence, however, of certain
substances when diluted, and the fact that disinfectants are some-
times equally efficacious in a diluted form when their application is
prolonged, seem to indicate measures which may be adopted, in some
cases, with chances of success, such as the inhalation of antiseptic
vapours in phthisis. For the most part the physician must look
rather to combating the effects of micro-organisms by restoring to
its normal standard the lowered vitality which enabled the bacteria
to get a footing.
32 BACTERIOLOGY.
There is no wider field for research than the determination of
the real effect of disinfectants and antiseptics. Painstaking and
laborious as the researches are which have been hitherto made, the
subject is so beset with fallacies, leading, in some cases, to totally.
erroneous conclusions, that it is not surprising that one meets on
all sides with conflicting statements. The author has no intention
of analysing these results, but a general idea will be given of the
methods which have been employed, and for further details reference
must be made to the original papers mentioned in the bibliography.
Chemical Substances.—It was customary to judge of the power
of a disinfectant or antiseptic by adding it to some putrescent liquid.
A small portion of the latter was, after a time, transferred to
some suitable nourishing medium, and the efficacy of the substance
estimated by the absence of cloudiness, odour, or other sign of
development of bacteria in the inoculated fluid. Koch pointed out.
the errors that might arise in these experiments from accidental
contamination, or from there being no evidence of the destruction
of spores, and we are indebted to him for a complete and careful
series of observations with more exact methods.
Instead of employing a mixture of bacteria, Koch’s plan was to
subject a pure cultivation of some well-known species with marked
characteristics to the reagent to be tested. A small quantity was _
then transferred to fresh nourishing soil, under favourable con-
ditions, side by side with nutrient material inoculated from a
cultivation without treatment with the disinfectant. The latter
constituted a control test, whichis most essential in all such
experiments. To test the resistant power of bacteria which are
easily destroyed, two species were selected, Micrococcus prodigiosus,
and the bacillus of blue pus. These were cultivated on potatoes,
the surfaces of which were sliced off and dried. A fragment trans-
ferred to freshly prepared potato gave rise to a growth of the
particular micro-organism ; but if after treatment with some reagent
no growth occurred, the conclusion was drawn that the reagent was
efficacious in destroying the vitality of the bacteria.
Anthrax bacilli in blood, withdrawn from an animal just killed,
were taken to represent sporeless bacteria, while silk threads steeped
in an artificial cultivation of the bacilli and dried, afforded a means
of testing the vitality of spores.
Even by employing pure cultivations on solid media, great
precautions were necessary to avoid mistakes. When, for instance,
a large quantity of the growth which had been subjected to some
chemical solution was carried over to the fresh tube containing
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 33
the nutrient medium, or when a silk thread, which had been dipped
in a solution, was directly transferred to the new soil, enough of the
supposed disinfectant might be mechanically carried over to retard
the development of the bacteria, though it was ineffectual in
destroying them. From a growth not appearing, it was concluded
that the spores or the bacteria had been affected, and so a
mistake occurred. To avoid this, Koch made a point of transfer-
ring a minimum of the disinfected growth to as large a cultivation
area as possible, so that any chemical substance mechanically
carried over would be so diluted as to be inert. For the same
reason, threads, after withdrawal from the disinfecting solution,
were rinsed in sterilised water, or weak alcohol, and then trans-
planted ; or, instead of judging from. the development on nutrient
gelatine, the effect of inoculation in a healthy animal was made
the test.
A few examples may be quoted in illustration. Silk threads,
impregnated with anthrax spores, were placed in bottles containing
carbolic acid of various strengths. A thread was removed from each
on successive days, and transferred to nutrient gelatine, and the
result noted. Jt was found that immersion of the thread in a 5 per
cent. solution of carbolic acid was sufficient in two days to effect
complete sterilisation, and seven days in a 3 per cent. solution was
equally efficacious. Since for practical purposes a strength should
be selected which would be effectual in twenty-four hours, Koch
recommended that for general use, allowing for deterioration by
‘keeping, a solution containing not less than 5 per cent. should be
employed, and for complex fluids probably a still higher percentage
would be necessary. In the case of sporeless bacilli the results were
very different. Blood containing the bacilli, from an animal just
killed, was dried on threads, and after exposure for two minutes to
a 1 per cent. solution, was completely sterilised; and fresh blood
mixed with a 1 per cent. carbolic solution produced no effect when
inoculated. On the other hand, when the blood was mixed with a
‘D per cent. solution, the virulence was not destroyed. The facility
with which the bacilli are destroyed, compared with their spores,
illustrates how easily errors may occur, when mere arrest of growth
or loss of motility is regarded as a sign of the efficacy of disinfection.
To test vapours, Koch exposed anthrax spores or the spores
which occur in garden earth by suspending them over solutions,
such as bromine or chlorine, in a closed vessel. After a time they
were transferred to a nutrient medium to test their vitality. To
test the power of sulphurous acid gas, the spores were spread about
3
34 BACTERIOLOGY.
in a room in which the gas was generated by burning sulphur in
the ordinary way for disinfecting a room. To test chemicals which
inight be recommended for disinfecting vans and railway carriages,
spores were laid on boards, which were then washed or sprayed, and
the spores then transferred to the nutrient gelatine.
Sternberg has also made an elaborate series of experiments with
regard to the action of germicides. In this case cultivations of.
well-known pathogenic organisms in liquid media were employed.
The supposed germicide was added to the liquid cultivation, and
after two hours a fresh flask of sterilised culture was inoculated from
the disinfected cultivation, and placed in the incubator, In twenty-
four to forty-eight hours, if the chemical proved inefficient, there
was evidence of a growth of bacteria. Blyth has investigated the
disinfection of cultivations of Bacterium termo, of sewage, and
typhoid excreta, and, in conjunction with Klein, the effect of well-
known disinfectant materials on anthrax spores. Miquel, Laws,
and others have also contributed to our knowledge of the. effect
of antiseptics and disinfectants upon micro-organisms. In spite of
all that has been done there is room for many workers; a great
deal of ground must be gone over again to rectify discrepancies,
examine conflicting results, and thus determine what observations
may be relied upon for practical application.
This may be illustrated by referring in detail to some experiments
made with corrosive sublimate. Koch investigated a long list of
chemical reagents, and according to these experiments the salts of
mercury, and the chloride especially, proved most valuable. Where
heat is not admissible, these compounds were therefore highly
recommended, though their poisonous nature is a drawback to their
indiscriminate use. Koch stated that for disinfecting a ship’s bilge,
where a 5 per cent. solution of carbolic acid must be left forty-eight
hours, a 1 in 1000 solution of mercurie chloride would only require
a few minutes.
But there was good reason for doubting the efficacy of very dilute
solutions; for, though according to Koch’s experiments anthrax
spores subjected to a 1 in 20,000 solution of mercuric chloride for
ten minutes, and then washed in alcohol, gave no growth in nutrient
gelatine, silk threads exposed for ten minutes to a 1 in 20,000
solution, or even 1 in 10,000, still proved fatal to mice.
Herroun cultivated ordinary septic bacteria in albuminous
filtrates, containing .1 in 2000, and concluded that the value of
mercuric chloride as an antiseptic was much over-rated. It is pre-
cipitated by albumins though, as Lister has shown, the precipitate
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 39
of albuminate of mercury is redissolved when there is an excess of
albumin present.
Geppert, and later Behring, recognised that the methods employed
for testing the efficacy of corrosive sublimate were unreliable. They
found, for example, that corrosive sublimate could not be removed
from silk threads by washing; and therefore to study the effect of
this antiseptic acting for a given time, it was necessary to dip the
threads in ammonium sulphide solution after the treatment with
corrosive sublimate.
The author confirmed the results of Geppert and Behring, and
made a series of experiments to test the value respectively of carbolic
acid and corrosive sublimate in antiseptic surgery. The method
of dipping an infected thread into an antiseptic solution for a few
uiinutes, and then transferring it to the surface of a nutrient medium
to test its efficacy in a given time, was discarded as-fallacious; the
thread being still wet with the solution when transferred to the
medium, it was obvious that the action of the antiseptic continued
for many days, To wash infected silk threads with alcohol after
exposure to the antiseptic to stop its further action, also proved to
be a fallacious method, for the author found in control experiments
that absolute alcohol will destroy Streptococcus pyogenes, erysipelatis,
and Staphylococcus pyogenes aureus, acting for only one minute.
Other methods were therefore resorted to, and cultures on the
sloping surface of nutrient agar were at first used. The antiseptic
was poured into the culture tube until the growth was covered,
and when it had acted for a definite time (one minute, five minutes,
or fifteen minutes) a solution was added which immediately stopped
further action. In the case of corrosive sublimate, ammonium sul-
phide was employed, which is quite inert as anantiseptic. The liquid
contents of the test tube were carefully poured off, and an inoculation
was made into a fresh tube of broth or agar from the culture still
adhering to the surface of the nutrient medium. As the results
disproved the efficacy of corrosive sublimate, it was thought possible
that the solution had not been able in the time to penetrate the
film of growth, Another plan was accordingly adopted. Cultures
were made in broth, and when fully developed the supernatant liquid
was carefully poured off. Corrosive sublimate solution was added to
the test. tube, and agitated until any flocculent masses were dis-
integrated and the whole of the liquid became uniformly turbid.
Ammonium sulphide was added when the time had expired, and
tubes of fresh broth were inoculated with the mixture. In the case
of carbolic acid the cultures, after its action, were thoroughly washed
36 BACTERIOLOGY.
with water, and its efficacy tested by making inoculations from the
cultures in fresh media. The results were entirely in favour of
carbolic acid. Staphylococcus pyogenes aureus and Streptococcus
pyogenes were not destroyed, even when corrosive sublimate solution
of 1 in 1000 was allowed to act for an hour. In the case of the
cultures of streptococcus of erysipelas the results were different.
A solution of 1 in 10,000 had no effect, but 1 in 4,000, acting for
one minute, destroyed the culture. With carbolic acid the results
were very striking. Cultures were exposed to solutions of 1 in 20,
1 in 30, 1 in 40, 1 in 50, for one minute, five minutes, fifteen
minutes. The attempts to make subcultures in every case failed.
Carbolie acid 1 in 40, acting for only one minute, was sufficient to
destroy Streptococcus pyogenes and Streptococcus erysipelatis and
Staphylococcus pyogenes aureus. Further experiments were made
with tubercular sputum, the test being subsequent inoculation of
guinea-pigs. Corrosive sublimate solution as strong as 1 in 500 had
no effect, but 1 in 20 carbolic acid, shaken up with the sputum for
one minute, completely neutralised it.
Koch’s statements with reference to the germicidal power of
corrosive sublimate in extremely weak solutions had led Lister to
substitute it for carbolic acid as a detergent in surgery. The author’s
experiments, which were undertaken in 1892, encouraged Lister to
revert to the use of carbolic acid, which, indeed, had always proved
efficacious in surgical practice. Lister pointed out that carbolic acid
has also the great advantage of combining eagerly with fats and
epidermis, so that the seat of operation can be effectually cleansed.
These experiments also point to the conclusion that carbolic acid
should be used in hospital wards for the disinfection of tubercular
sputum instead of mercuric chloride and other less efficacious dis-
infectants commonly in use.
Hot Air and Steam—Koch, in conjunction with Wolfhiigel, also
made exhaustive experiments to test the value of hot air. A similar
plan was adopted to that employed in disinfection with chemicals.
Bacteria and spores were subjected for a certain time to a known
temperature in the hot-air chamber, and then were transferred to a
nourishing soil or inoculated in animals.
Paper parcels, blankets, bags, and pillows, containing samples of
micro-organisms wrapped up inside, were also placed in the hot-air
chamber, to test the power of penetration of heat.
The conclusions from these experiments were as follows :—
Sporeless micro-organisms at a little over 100° GC. are destroyed
in one hour and a half,
EFFECT OF ANTISEPTICS AND. DISINFECTANTS ON BACTERIA. 37°
Spores of bacilli require three hours at 140° C.
If enclosed in pillows and blankets, exposure from three to four
hours to 140° C. is necessary.
Spores of fungi require one and a half hours at 110° 0. to
115° C.
Further experiments showed that at the temperature necessary
for the destruction of spores of bacilli almost all fabrics are more or
less injured.
Koch, in conjunction with Gaffky and Léffler, also investigated
the effect of steam under pressure and at the atmospheric pressure.
Rolls of flannel with anthrax spores or earth spores, and a
thermometer wrapped up inside, were subjected to steam, and
the results compared with the effect obtained with hot air.
Thus in hot air four hours’ exposure to a temperature of 130° C.
to 140° C. brought the temperature inside the roll to 85° C., and the
spores were not injured; on the other hand, exposure to steam
under pressure at 120° C. for one and a half hours, raised the
internal temperature to 117° C. and killed the spores.
By such experiments the superior penetrative power of steam-
heat was established.
To test steam-heat at the atmospheric pressure, water was boiled
in a glass flask with its neck prolonged by means of a glass tube, the
temperature in which was found to be uniform throughout. Anthrax
and earth spores placed in the tube were found to be unable to with-
stand steam at 100° C. even for a few minutes. It was, therefore,
concluded that disinfection by steam at atmospheric pressure was
superior to hot air from its greater efficiency, and to steam under
pressure from the simplicity of the necessary apparatus.
Parsons and Klein made some experiments which were more
in favour of dry heat than the above. These observers state that
anthrax bacilli are destroyed by an exposure of five minutes at
from 100° C. to 103° C. and that anthrax spores are destroyed
in four hours at 104° C., or in one hour at 118° C. Guinea-pigs
inoculated with tuberculous pus which had been exposed for five
‘minutes to 104° C., remained unaffected. They concluded that as
none of the infectious diseases, for which disinfecting measures
are in practice commonly applied, are known to depend upon the
presence of bacilli in a spore-b2aring condition, their contagia
are not likely to retain their activity after being heated for an
hour to 105° C. (220° Fahr.)
In experiments with steam the results were in accordance
with those already given, and complete penetration of an object
38 BACTERIOLOGY.
by steam-heat for more than five minutes was deemed sufticient.
They also arrived at the same result as in Koch’s experiments,
viz., that steam-chambers are preferable to those in which dry heat
is employed, though it must be borne in mind that some articles,
such as leather, are injured by exposure to steam.
PracticaL APPLICATION.
Nurses and others attending infectious cases should freely use
1 in 40 carbolic for the hands and a weaker solution for the body
generally, The skin of patients after recovery should be sponged
with 1 in 40 carbolic. The dead should be wrapped up in a sheet
soaked in 1 in 20 carbolic acid or a strong solution of chloride of
lime. Infected clothing and bedding should be burnt unless in excep-
tional cases, when they may be disinfected by boiling, or by exposure
to dry heat at 105°C. to 110°C. for three hours, or by steaming
at 100°C. for fifteen minutes. Leather and other articles which
would be destroyed by any of these processes should be sponged with
lin 40 carbolic. The walls of the sick-room and furniture should
be exposed to the fumes of burning sulphur, and next day washed
down with 1 in 40 carbolic, and the room freely ventilated by
opening all windows and doors. Rags should be burnt, or dis-
infected by boiling or exposure to steam when supplied to manu-
facturers. _The importation of rags from places where there are
eases of cholera or small-pox should be prohibited. Infected ships
must be fumigated with sulphur, and the bilge disinfected with
carbolic acid. Infected railway carriages should be disinfected in
the same way as a sick-room.
Tubercular sputum, cholera and typhoid evacuations and other
excreta should be disinfected by 1 in 20 carbolic acid, or by a strong
solution of chloride of lime.
CHAPTER IV.
CHEMICAL PRODUCTS OF BACTERIA.
Tue products of the metabolism induced by bacteria may be divided
into three classes : (1) ptomaines or alkaloids; (2) albumoses or tox-
albumins ; and (3) enzymes, Alkaloids and albumoses are directly
poisonous ; enzymes or ferments are harmless except in the presence
of proteids, which they are capable of transforming into poisonous
albumoses.
ProMAInges AND Tox-ALBUMINS.
The study of these products may be said to date back to 1822)
when Gaspard and Stick found an intensely poisonous principle
in cadaverous extracts. In 1856 Panum discovered « poisonous
substance in putrid flesh; and in 1863 Bergmann and Smiedeberg
found a nitrogenous crystallisable substance in putrid beer which
they named sepsin. In 1872 Gautier found that the decomposition
of fibrine led to the formation of various complex alkaloidal sub-
stances, and in 1875 Richardson obtained in pyemia an alkaloid,
septin. This subject, however, received most attention from the
classical researches of Selmi, the Italian toxicologist. Selmi, in a
celebrated poisoning case, demonstrated the presence of an alkaloid
as the result of post-mortem changes. Similar substances were
found in alcohol in which morbid specimens had been preserved.
Thus the researches of Gautier and Selmi established the fact that
albuminoid material undergoing decomposition leads to the forma-
tion .of cadaveric alkaloids. These animal alkaloids Selmi named
ptomaines. Brieger, finding the bases derived from the products of
putrefaction less poisonous than those obtained from the pathogenic
bacteria, suggested the term toxins for the latter. Ptomaines have
been divided into two classes—those which are non-oxygenous, liquid,
and volatile, and those which are oxygenous, solid, and crystallisable.
They are, for the most part, precipitated by the ordinary reagents
39
40 BACTERIOLOGY.
for alkaloids, such as chloride of gold, double iodide of mercury
and potassium, picric acid, and tannin. Phospho-molybdie acid
precipitates them without exception. They are powerful reducing
agents. Ferro-cyanide of potassium is converted into ferri-cyanide in
their presence, and the addition of ferric chloride gives the Prussian
blue test. Selmi discovered this test, and Brouardel and Boutmy
regarded it as absolutely characteristic of ptomaines ; but this is
not the case; some vegetable alkaloids, for example, behave in the
same way.
As examples of the non-oxygenous ptomaines there are :—
Parvolin (C9H¥N) an oily base of an amber colour prepared
from putrid mackerel and horse-flesh.
Hydrocollidin (C83H4*N), from the same source, It is highly toxic,
being compared by Gautier to the venom of the cobra di capello.
Collidin (C8HUN), from putrid gelatine and the pancreas of a
bullock, also highly toxic.
Neuridin (C@HN?), from fish, flesh, and decaying cheese.
Saprin (C6HMN?), isomeric with neuridin.
Cadaverin (CH™N?), a third isomeride, from ordinary putrefac-
tion and herring brine.
Putrescin (C4H2N?) from putrefaction.
The oxygenous ptomaines are in some instances found also in
healthy tissues. They include the following :—
Newrin (C5H4NO), found in cadaveric putrefaction.
Cholin (C*H5NO’), in bile.
Muscarin (COHBNO?), in a poisonous mushroom, Agaricus mus-
carius, and in putrid fish. These are all highly poisonous.
Gadinin (CTHISNO?), in putrefying codfish.
Mytilotoxin (COH™NO?), in poisonous mussels.
Poisonous alkaloids are of great importance in connection with
those cases of meat poisoning produced by sausages, hams, poultry,
and cheese. Tyrotoxicon is a poisonous alkaloid obtained from cheese.
The toxic substances of most interest to the bacteriologist are
those isolated from pure cultivations of pathogenic bacteria, such as
typhotoxin, isolated by Brieger from cultivations of the bacillus of
typhoid fever, and tetanin, from cultivations of the tetanus bacillus ;
and the poisons known as albumoses or tox-albumins, which are
allied to the albumose of snake poison.
Pasteur, in 1885, suggested that in anti-rabic inoculations the
immunity resulted from the action of a substance secreted by a
microbe, though the microbe has not as yet been discovered in
rabies. Salmon produced immunity from hog cholera by the injec-
CHEMICAL PRODUCTS OF BACTERIA. 4]
tion of the toxic products in filtered culture fluids. Wooldridge,
Hankin, and Martin studied the products of Bacillus anthracis.
Charrin, and later Woodhead, Wood, and Blagovestchensky, investi-
gated on these lines Bacillus pyocyaneus. Roux and Chamberland
experimented with the bacillus of malignant edema; Roux with
symptomatic anthrax; Chantemesse and Widal with the typhoid
bacillus. Roux, Yersin, Brieger, Frankel, Martin, and Behring
worked on the same lines with diphtheria. Koch introduced
tuberculin, Kalning mallein, while others have utilised the products
of streptococci and pneumococci. Anrep found an albumose in the
medulla of rabid animals, and Babés claims to have found an
albumose in both rabies and glanders.
Cholera.—Brieger found several ptomaines, including putrescin
and cadaverin, in pure cultures of the spirillum of Asiatic cholera,
and Petri found in addition to poisonous bases a proteid body which
produces in guinea-pigs muscular tremors, paralysis, and a rapidly
fatal result. Roux and Yersin obtained from cultures a tox-albumin
insoluble in water, which kills guinea-pigs in two or three days,
but has no effect on rabbits. Pfeiffer also investigated the toxic
substances in cultures. Chloroform, thymol, and drying destroyed
comma-bacilli, leaving their toxic products unaffected. Concentrated
solutions of neutral salts and boiling produced secondary toxic
substances, but the original toxic substances were ten or twenty
times more virulent.
Typhoid Fever.—Typhotoxin (C7H!7NO?), the alkaloid ob-
tained by Brieger from cultures of the typhoid fever bacillus, produces
in mice and guinea-pigs salivation, rapid breathing, dilatation of the
pupil, diarrhea, and death in twenty-four to forty-eight hours. At
the post-mortem examination the heart is found in a state of systolic
contraction, and the condition of the heart after death and the
absence of convulsions during life serve to distinguish typhotoxin
from an isomeric base obtained by Brieger from putrid horse-flesh.
Roux and Yersin have obtained a tox-albumin. It is soluble with’
difficulty in water, and more toxic to rabbits than guinea-pigs.
Tetanus.—Brieger obtained the alkaloid tetanin from impure
cultures of the tetanus bacillus. It is a base having the formula
C3H22N?04, The hydrochloride is a very deliquescent salt, and
soluble in alcohol. Tetanin injected into guinea-pigs produces
rapid, breathing, followed by tetanic convulsions. Another toxic
product, tetanotoxin (C5H™UN), produces the same effects as tetanin.
The formula of a third base, spasmotoxin, has not been determined.
Cadaverin and putrescin are also present in cultures. Kitasato and
42 BACTERIOLOGY.
Weyl analysed the products of pure-cultures, and obtained the same
substances, tetanin and tetanotoxin ; and subsequently Brieger and
Frankel found that in pure-cultures a tox-albumin could be
obtained which is soluble in water, and infinitely more active ‘than
the toxic ptomaines.
Anthrax.—-In 1887 Wooldridge succeeded in protecting rabbits
from anthrax by a new method. A proteid body obtained from the
testis and from the thymus gland was used as. the culture fluid.
This proteid substance was dissolved in dilute alkali, and the solution
sterilised by repeated boiling. This. was inoculated with the anthrax
bacillus, and kept at 37° C. for two or three days. A small quantity
of the filtered culture fluid injected into the circulation in rabbits
produced immunity from anthrax. Subcutaneous inoculation of
extremely virulent anthrax blood, made simultaneously with the injec-
tion of the protecting fluid, produced no effect. Wooldridge showed
that the growth of the anthrax bacillus in special culture fluids
gave rise to a substance which, when injected into the organism,
protected not only against an immediate but also subsequent attacks.
In 1889 Hankin worked under the guidance cf Koch in the
Hygienic Institute of Berlin. The acquired tolerance of the effect
of ordinary albumoses, and the experiments of Sewall, who pro-
duced immunity against lethal doses of the albumose of snake
poison by the injection of minute doses, led Hankin to expect that
an albumose developed in anthrax cultures, ard that the anthrax
albumose would probably confer immunity from the disease. Hankin
succeeded in isolating it from culture fluids. It was precipitated by
excess of absolute alcohol, well washed in alcohol to free it from
addition of ptomaines, filtered, dried, then redissolved and filtered
through a Chamberland filter. With this substance Hankin suc-
ceeded in producing immunity in mice and rabbits.
Sidney Martin, working quite independently, grew anthrax bacilli
in a solution of pure alkali albumin made from serum proteids. After
ten or fifteen days the organisms were removed by filtration through
a Chamberland filter. The filtrate contained proto-albumose and
deutero-albumose, a trace of peptone, an alkaloid, and small quantities
of leucin and tyrosin. The mixture of albumoses proved poisonous
to mice. The anthrax alkaloid produced symptoms and lesions
similar to the albumoses, but much more rapidly and severely. It
is an amorphous yellow body, soluble in alcohol and alkaline in
reaction. Martin concluded that the anthrax bacillus formed the
albumoses and the alkaloid by digesting the alkali albumin; and
suggested that the alkalinity of the albumoses explained their toxi¢
CHEMICAL PRODUCTS OF BACTERIA. 43
properties, the alkaloid probably being in a nascent condition in the
albumose molecule.
Tuberculosis.—Koch prepared a glycerine extract of the
product of the tubercle bacillus in pure cultivations, and found
that the injection of small doses produced a remarkable reaction,
both local and general, in tubercular cases, and especially lupus.
This extract, called tuberculin, came to be extensively used as a
therapeutic agent, but with disappointing results. Very shortly
after the first announcement of Koch’s discovery, the author, in
conjunction with Herroun, investigated the chemical properties and
physiological effects of the products of the tubercle bacillus.
Cultures in glycerine-broth were filtered through porcelain, and
a clear amber-coloured liquid was obtained, which gave important
and suggestive chemical reactions. As this filtrate contained the
products of the growth of the bacillus most probably in minute
quantities, it was evaporated at «a low temperature over sulphuric
acid. The viscous residue was dissolved in distilled water and
tested on the healthy guinea-pig. The result was a marked fall of
temperature, staring coat, extreme irregularity of the heart’s action,
muscular spasms, loss of control over the extremities, and death.
A preliminary examination of glycerine-broth cultivations having
shown the presence of non-coagulable proteid bodies of the nature
of albuthose and peptone, and a crystallisable precipitate of a
remarkable character resulting on the addition of iodine, the idea
naturally suggested itself that the tubercle bacillus might form
albumoses and an alkaloid or ptomaine similar to the substances
isolated by Martin from pure cultivations of the Bacillus anthracis.
Koch pointed out that the effective substance in his extract
could be precipitated by absolute alcohol; the author and Herroun
determined to investigate the properties and physiological effects
of the separated products. They accordingly set to work to isolate
the ptomaine, of the existence of which they had some qualitative
indication, and at the same time to examine the properties of the
albuminous bodies.
In this endeavour the general method they found satisfactory was
as follows. The clear filtrate from the culture was evaporated at
40° ©. to a very small bulk, and the residue thus obtained was
mixed with an excess ef absolute alcohol, which precipitated the
albumoses and peptone. Tt was found that by adding the alcohol
by degrees a partial separation of the albumose from the peptone
could be effected, the latter being only precipitated when the aleohol
was nearly absolute. The precipitated albumose was collected on
44 BACTERIOLOGY.
a filter and redissolved in distilled water. In another experiment
the albumose underwent a second precipitation, and after washing
was again dissolved.
The alcoholic filtrate from the precipitated albuminous bodies was
then concentrated at a very gentle heat until a viscous residue was
left containing the glycerine originally present in the cultivating
medium and the extractives and products of the bacillus soluble
in alcohol. With this residue definite reactions of an alkaloidal
substance or ptomaine were obtained.
Careful experiments, however, led to the belief that the whole
of the ptomaine was not separated from the albuminous precipitate
by simple addition of alcohol, and the above method was therefore
slightly modified.
The ptomaine is soluble in water and alcohol, and sparingly
soluble in amyl-alcohol, but insoluble in benzine, ether, or chloro-
form, which liquids therefore fail to extract it from aqueous
solutions. In its aqueous solutions it is distinctly but not strongly
alkaline to test-paper. Phospho-tungstic acid gives with it a white
flocculent precipitate. Phospho-molybdic acid gives a pale yellow
precipitate, soluble in ammonia to a blue solution which becomes
colourless on boiling. In this respect it resembles the vegetable alka-
‘ loids, aconitin and atropin. It must be remembered, however, that
albuminous bodies are precipitated by both this and the preceding
reagents, and in the case of the former a reduction of the phospho-
molybdate giving the blue solution with ammonia is obtained.
The reducing power of the ptomaine is shown by the conversion
after a short time of ferri-cyanide of potassium to ferro-cyanide,
giving the Prussian blue test with ferric chloride, to which much
undue importance was attached by Brouardel and Boutmy. The
solution of albumose and solution of peptone are both capable of
giving this reaction as well as many vegetable alkaloids. A solution
of the ptomaine is not precipitated by ferro-cyanide of potassium or
potassic bichromate.
In strong solutions it yields precipitates with platinic chloride
(yellow), gold chloride (pale yellow), and mercuric chloride (white),
That yielded by the first of these reagents is granular in character,
and quite insoluble in alcohol, though apparently soluble in water.
The precipitation by gold chloride excludes amides and ammonium
salts.
With iodine in hydriodic acid or potassic iodide a precipitate is
obtained which is occasionally erystalline, more often granular or
amorphous.
CHEMICAL PRODUCTS OF BACTERIA. 45
This precipitate is soluble in alcohol, and is redeposited when the
alcohol is evaporated. On heating it is redissolved into oily drops of
a dark colour. .With picric acid a granular precipitate is obtained,
which under the microscope is seen to consist of minute crystals.
This precipitate, on standing, is converted into rounded crystalline
masses with numerous small crystals admixed.
The ptomaine appears to be easily broken up by heating,
especially in the presence of mineral acids or of baryta. The actual
quantity obtained from a considerable amount of culture fluid was
very small, and as it was possible that when the bacilli were grown
in a medium richer in albumin, such as the animal body, more of
these products might be formed, the liquid obtained by extracting
large masses of tubercular growths from cattle was examined in a
similar manner. In this extract, after filtration through porce-
Jain, an albumose, and minute quantities of a ptomaine were
obtained which in reactions was identical with that obtained from
the artificial cultivation of the bacillus, but present in even smaller
amount. The probable explanation of this is, that. in the living
animal the ptomaine is constantly being removed ; or it may indicate
that it is only formed in minute quantity under those conditions.
Having succeeded in obtaining the albumose and the ptomaine
in separate solutions, we next proceeded to ascertain the effects of
these substauces upon healthy and tubercular guinea-pigs.
The effect of the ptomaine isolated from different series of
cultures was as follows. A rise of temperature occurred in tuber-
cular animals, and distinct enlargement of tubercular glands. There
was a slight indication of a depression of temperature or hypothermic
effect on healthy animals. The albumose, whether obtained from
pure cultivations of the bacillus or from tubercular tissue, pro-
duced a marked rise of temperature in tubercular guinea-pigs. On
the other hand, in a control experiment on a healthy guinea-pig
there was an equally well-marked fall of temperature. The effect
upon the tubercular glands in the cases associated with marked rise
of temperature was to render them well-defined, indurated, and
painful, rather than any considerable increase in volume.
Hunter made a chemical examination of Koch’s crude extract,
and confirmed the presence of albumoses and alkaloidal substances.
The albumoses consisted chiefly of proto-albumose and deutero-albu-
mose with hetero-albumose, and occasionally a trace of dys-albumose.
Two alkaloidal substances were obtained in the form of platinum
compounds of their hydrochlorate salts. In addition there were
extractives, mucin, inorganic salts, glycerine, and colouring-matter.
46 BACTERIOLOGY.
Swine Fever.—Schweinitz applied Brieger’s methods in the
investigation of the products of the swine fever, or hog cholera
bacillus. Broth-cultures were neutralised with dilute hydrochloric
acid, and “evaporated in the water bath. The residue was treated
with 96 per cent. alcohol, and the filtered solution with mercuric
chloride. A heavy crystalline precipitate was separated by filtra-
tion, treated with water, and decomposed with sulphuretted hydrogen,
and cadaverin and methylamine were isolated. The filtrate from
the mercuric chloride precipitate was freed from excess of mercury
by sulphuretted hydrogen, and the mercury sulphide filtered off.
The residue, after concentration of the filtrate, was extracted with
absolute alcohol, and the solution showed the presence of an alka-
loidal salt. The double salt obtained with platinum chloride was
submitted, after crystallisation, to an analysis, and the results gave
the formula (C4H*4N?PtClS). The hydro-chloride is soluble in abso-
lute alcohol as well as in water, and produces needle-like crystals.
On treating the culture fluids with excess of absolute alcohol a
white flocculent precipitate was obtained partly soluble in water, and
re-precipitated by alcohol. It was obtained in the form of white
crystalline plates. A watery solution gives almost insoluble needle-
crystals on the addition of platinum chloride. These products were
respectively termed sucholo-toxin and sucholo-albumin. Small doses
of these substances produce in guinea-pigs a slight rise in tempera-
ture, and ulceration at the seat of injection. Large doses produce
a fatal result in six to twenty-four hours. Schweinitz asserts that
he has produced immunity in guinea-pigs. An attempt to produce
immunity in swine by injection of the albumose gave unsatisfactory
results.
Diphtheria.—Roux and Yersin finding that filtered cultures of
the diphtheria bacillus produced paralysis, affecting chiefly the hind
legs, and a fatal result in rabbits and guinea-pigs, proceeded to
investigate the chemical products. They succeeded in obtaining a
white amorphous substance which was extremely active when
injected into guinea-pigs. It was precipitated by alcohol from an
aqueous solution, and it was calculated that -0004 gram would
destroy eight guinea-pigs of 400 grams, or two rabbits of 3 kilos.
each. They concluded that the poison was an enzyme or ferment,
as it not only acted in extremely small doses, but it was attenuated
by heat and destroyed by boiling.
Brieger and Frinkel confirmed these experiments, and asserted
that the poison was a tox-albumin; but according to Martin their
chemical analysis and reactions were vitiated by the fact that they
CHEMICAL PRODUCTS OF BACTERIA. 47
had peptone in their cultivating medium. Martin examined the
products by using as a culture medium a 1 to 2 per cent. solution
of alkali-albumin in broth made from beef, omitting the peptone.
After about thirty days the bacillus had converted the alkali-albumin
into albumoses, which gave the reactions of proto- and deutero-
albumose, with small quantities of an organic acid. A single dose
of these albumoses produced weakness of the hind limbs, which after
a time passed off. The animal was killed, and the nerves which
were examined showed degeneration. Repeated intravenous in-
jection on successive days, amounting in all to a dose of 1:69 grams
per kilo. of body weight, produced high fever, followed by depres-
sion of temperature, severe watery diarrhea, and emaciation. The
tendon reflexes began to diminish after the ninth day, on the
eleventh or twelfth day there was definite paralysis of the hind legs,
and on the seventeenth day reflexes could scarcely be obtained.
Martin thus gives his method of abstracting the poisonous pro-
ducts either from cultures or from diphtheritic tissues. In dealing
with tissues, the spleen and other organs are first finely minced and
placed in rectified spirit, and the blood is also placed in spirit, and
allowed to stand till the proteids are coagulated; they are then
filtered, and the residue extracted with cold water, all the extracts
are mixed together, and evaporated at 35° C. tv a small bulk, and
thrown into absolute alcohol. Most of the albumoses are precipi-
tated, the alcohol is poured off, evaporated to dryness at a low
temperature, and extracted by absolute alcohol until nothing more
dissolves. The residue is deutero-albumose and mineral salts. All
the proteid is mixed together, dissolved in water, and precipitated by
alcohol, the process being repeated to remove any traces of bodies
soluble in alcohol and the excess of mineral salts. At the last
precipitation the precipitate is allowed to stand under alcohol for
about two months. The alcohol is then poured off, and the pre-
cipitate dried im vacuo.
The resulting product is a light yellowish-brown powder soluble
in water, cold or boiling, giving a yellowish and faintly acid or
nearly neutral reaction. It is composed of deutero-albumose with
a slight amount of proto-albumose but no peptone. It gives the
ordinary actions of proteids and a well-marked biuret reaction. It
is precipitated from solution by ammonium sulphate, and slightly
by nitric acid. The reactions are similar to those of peptic deutero-
albumose. The alcoholic extract of the tissues is strongly acid, and
contains free fatty acid and an organic acid insoluble in chloroform.
The organic acid is readily soluble in water and absolute alcohol,
48 BACTERIOLOGY.
and insoluble in ether, chloroform, and benzine. It is a yellowish
amorphous body, becoming a deep brown when made alkaline.
Martin concludes that whereas the Bacillus anthracis produces
albumoses and an organic base, in diphtheria we find albumoses and
an organic acid,
Glanders.-—Kalming has obtained from cultures of the glanders
bacillus an extract similar to tuberculin. This crude extract is
known as mallein, and is extensively used for the diagnosis of
glanders. In a glandered horse it causes a rise of temperature
and swelling at the seat of the injection, and the glandered nodules
become swollen and painful. Finger claims to have produced
immunity from glanders by inoculation of the products contained
in sterilised cultures. Schweinitz extracted from cultures a non-
poisonous albumose, and obtained only traces of a ptomaine.
Suppuration and Pneumonia.—Brieger obtained a ptomaine
from cultures of Staphylococcus pyogenes aureus, and Roux and
Yersin a tox-albumin fatal to rabbits and guinea-pigs in a few
days. There was pus-formation at the seat of inoculation, with
redness and swelling of the surrounding parts.
From pure-cultures of the micrococcus of pneumonia Klemperer
obtained a tox-albumin, for which the name pneumo-toxin has been
suggested. .
Enzymes on FERMENTS.
Many bacteria liquefy the nutrient gelatine in which they are
cultivated. This is due to the development of a ferment or enzyme,
which dissolves the albumin and gelatine.
Enzymes are products of the vital activity of living bacteria.
Bitter, and independently Sternberg, showed that when a liquefying
bacterium is removed by filtration or destroyed by heat, the culture
fluid retains the power of liquefying gelatine. As this occurs
usually when the reaction is alkaline, bacterial enzymes resemble
trypsin and papain rather than pepsin. They can be extracted with
glycerine, and are quite harmless. If injected into animals no effect
is produced, and after a few hours no trace of them can be found.
According to Fermi, the influence of temperature on the enzymes
produced by different bacteria will be found to vary very consider-
ably. The enzyme of Staphylococcus pyogenes aureus is destroyed
at 55° C., while the enzyme of Bacillus anthracis succumbs at a
temperature of 65° C. to 70° C.
Some bacteria produce both enzymes and toxins, but many pro-
duce enzymes and not toxins, and others toxins but not enzymes.
CHAPTER V.
IMMUNITY.
THE condition of being insusceptible to an infective disease may be
either natural or acquired. In studying the pathogenic organisms
several examples of natural immunity will be encountered. The.
bacillus of septicemia, so fatal to house mice, has been shown to
have no effect upon field mice. The bacillus of anthrax is innocuous
to cats and white rats. The bacterium of rabbit septicemia is
equally inert in dogs, rats, and guinea-pigs. The immunity may
be as in these cases complete, or only partial. Ordinary sheep are
very easily affected with anthrax, but Algerian sheep succumb only
to large doses of the virus. Natural immunity may not only be
characteristic of certain species, but it may occur in certain indi-
viduals of a susceptible species. The same immunity occurs in man,
for certain individuals, though equally exposed during an epidemic
of small-pox, may escape, whereas others readily fall victims to the
disease.
Acquired immunity is illustrated by the protection afforded by
one attack of the exanthemata against subsequent attacks, Thus
one attack of measles or small-pox, as a rule, affords complete
protection. A knowledge of the immunity resulting in the latter
case led to the introduction of inoculation of small-pox as a
protection against natural small-pox,
Immunity may be acquired by acclimatization, for the inhabit-
ants of tropical climates are less susceptible to the diseases of the
country, malarial fevers, for instance, than strangers.
In civilised communities also, there appears to be a degree of
acquired immunity, for infectious diseases like measles introduced
among savages or isolated communities have assumed the most
malignant type.
The immunity acquired by protective inoculation constitutes, in
connection with the study of pathogenic micro-organisms, a subject
of pre-eminent interest and importance, Pasteur, in his researches
49 4
50 BACTERIOLOGY.
upon fowl-cholera, observed that after non-fatal cases the disease
either did not recur, or the severity of a subsequent attack was in
inverse proportion to the severity of the first attack. It occurred
to him to endeavour to obtain the virus of this disease in a form
which would provoke a mild attack of the disease, and thus give
protection against the virulent form. This attenuation or miti-
gation of the virus was successfully attained by allowing cultiva-
tions of the microbe in chicken-broth to remain with a lapse
of several months between the carrying on of successive cultiva-
tions in fresh media. The new generations which were then
obtained were found to have diminished in virulence, and ultimately
a viruS was obtained which produced only a slight disorder ; on
recovery the animal was found to be proof against inoculation with
virulent matter. The explanation given by Pasteur ofthis change
was, that prolonged contact with the oxygen of the air was the
influence which diminished the virulence, and he endeavoured to
prove this by showing that when broth was inoculated in tubes
which .could be sealed up, so that only a small quantity of air
was ‘accessible to the microbe, the virulence of the cultures was
retained.
' Toussaint investigated the possibility of attenuating the virus of
anthrax. Sheep injected with 3 cc. of defibrinated blood, con-
taining anthrax bacilli, which had been exposed to 55°C. for ten
minutes, recovered, and were afterwards insusceptible. Pasteur
subsequently argued that this method did not admit of practical
application, because difficulties would arise in dealing with infective
blood in quantity, and artificial cultivations started from this blood
could not be relied upon, as they proved sometimes as virulent as
ever.
Pasteur endeavoured to apply the same method for- obtaining
an attenuated virus of anthrax, as he had successfully employed
in fowl-cholera. A difficulty was soon encountered, for in culti-
vations of this bacillus, with free access of air, spore-formation
readily takes place, and the spores are well known to have an
extraordinary power of retaining their virulence. Pasteur found
that the bacilli ceased to develop at 45°C., and he believed that
spore-formation ceased at 42° to 43°C., the bacilli continuing to
develop by fission only. The cultivations were, therefore, kept at
this temperature, and at the end of eight days the bacilli were
found ‘to have ° lost their virulence, and were quite inert when
inoculated in guinea-pigs, sheep, or rabbits. This total destruction
was, however; preceded by a gradual mitigation; so- that a virus
IMMUNITY, 51
could be obtained, by taking it at the right time, which gave only a
mild disease, and afforded subsequent protection.
At Melun, in 1881, the protective inoculation against anthrax
was put to a practical test. Sheep and oxen were inoculated with
the mitigated virus, and then with a virulent form; at the same
time other sheep and oxen were inoculated with the virulent form
without previous vaccination, as a control experiment, The unpro-
tected sheep died without exception ; the unprotected oxen suffered
from cedematous swellings at the seat of inoculation, and a rise of
temperature ; but all the protected animals remained healthy.
As a result of these experiments an idea arose that by preventive
inoculation with attenuated virus all communicable diseases would
in time be eradicated ; but this does not follow, for all communi-
cable diseases do not confer immunity after a first attack;
influenza, the very reverse is believed to occur, and erysipelas of the
face leads to an increased liability to subsequent attacks. Even
with regard to the prevention of anthrax, Pasteur’s researches were
opposed and criticised. Koch investigated the subject, and came to
the conclusion that the process did not admit of practical applica-
tion, chiefly on the ground that as immunity lasted only a year, the
losses from the vaccination process would be as. great or even
greater than from the spontaneous disease ; further, there was
danger in disseminating a vaccine of the strength required to be
effectual.
Chauveau proved that the attenuation was due to the tempera-
ture, and not to the prolonged effect of oxygen. By keeping
cultivations at 42° to 43°C. im vacuo, the virulence was found
to disappear in twenty-four hours, and by keeping cultivations
at a low temperature with free access of air, the virulence was
retained. Chauveau considered, therefore, not only that oxygen was
not the agent, but that the mitigation was much more easily effected
in its absence. In spite of these adverse criticisms, these researches
nevertheless confirmed the principle of Pasteur’s conclusion, that
immunity could be induced by experimental measures, and further
showed that he had considerably advanced the methods by which this
could be effected.
Chauveau succeeded also in attenuating the virus by a modifica-
tion of Toussaint’s method. Sterilised broth was inoculated with
the bacilli, and placed in the incubator at 42° to 43°C. After the
lapse of twenty hours it was removed to another incubator at 47° C.
According to the time of exposure to this increased temperature, the
mitigation varied in degree. Thus inoculation with the virus,. before
52 BACTERIOLOGY.
it was exposed to 47°C., was fatal to guinea-pigs; but after one
hour at 47°C, the virulence was diminished, and, though ultimately
fatal, life was prolonged; after two hours’ exposure at 47° C. only
half the animals died; and after three hours’ exposure they
recovered, and were rendered refractory to subsequent inoculation,
Attenuation of the virus of anthrax has also been induced by
chemical means, Chamberland and Roux stated that a fresh growth
started from a cultivation of bacilli which had been subjected for
twenty-nine days to gj of carbolic acid was found to be inert in
guinea-pigs and rabbits. Bichromate of potash added to a cultiva-
tion in the proportion of ;sigy tO sono gave, after three days, a
new growth, which killed rabbits, guinea-pigs, and half the sheep
inoculated ; after ten days, rabbits and guinea-pigs, but not sheep;
and after a longer time even guinea-pigs were unaffected.
In other diseases similar results have been obtained. Arloing,
Cornevin, and Thomas found that by inoculating a small quantity
of the virus of symptomatic anthrax anywhere in the subcutaneous
connective tissue, or a moderate quantity at the root of the tail,
and even by intravenous injection, immunity was obtained from a
virulent dose.
In swine-erysipelas, Pasteur and Thuillier obtained attenuated
virus upon quite another principle. They discovered that by
passing the virus through pigeons the virulence was increased, but
by passing it through rabbits it was progressively diminished, Thus
a virus was obtained from the rabbit, which produced only a mild
disease in pigs, and after recovery complete immunity. Similarly
in rabies, Pasteur found that passage of the virus through various
animals considerably modified its properties, By inoculating a
monkey from a rabid dog, and then passing the virus through other
monkeys, the virulence was diminished ; but by inoculating a rabbit
from the dog, and passing the virus from rabbit to rabbit, the
virulence increased,
In rabies, Pasteur has employed another method of attenuating
the virus. The spinal cord of inoculated rabbits is removed with
all possible precautions, and portions a few centimetres in length
are suspended in flasks in which the air is dried by fragments of
potash. By this process the virulence is found to gradually diminish
and finally disappear. Animals inoculated with portions of these
cords, after suspension for a certain time, are rendered refractory to
inoculation with virulent cords. Having rendered dogs, which had
been previously bitten, free from the supervention of symptoms of
hydrophobia by means of protective inoculation, Pasteur proceeded
IMMUNITY. 53
to apply the same treatment to persons bitten by rabid animals, with
results which tend to the belief that a real prophylactic for rabies
has been discovered.
Immunity may also be produced by injecting the toxic products
existing in pure cultivations after removal of the bacilli. Salmon
was the first to produce immunity in this way, by utilising the toxic
products of the bacterium of hog-cholera, which were separated by
filtration from the living micro-organisms; and shortly afterwards
Wooldridge demonstrated that filtered anthrax cultures contained
a substance which conferred immunity. Behring and Kitasato
produced immunity by mixing cultures with terchloride of iodine.
Vaillard filtered the cultures through porcelain, and attenuated the
products by heating at different temperatures.
Lastly, in the course of Behring’s and Kitasato’s experiments, it
was found that the blood serum of animals rendered immune was
capable of conferring immunity on other animals. The injection
of the toxic products of pathogenic bacteria leads to the development
of substances in the blood to which the term “ antitoxin” has been
applied. These protective substances neutralise or destroy the
injected poison, and blood serum which has thus been rendered
antitoxic can be utilised to confer immunity on other animals.
Haffkine’s system of vaccination as a protection against Asiatic
cholera is supposed to be based upon the principle of inducing the
formation of antitoxins or defensive proteids.
MEcHANISM OF IMMUNITY.
Raulin has shown that Aspergillus niger develops a substance
which is prejudicial to its own growth, in the absence of iron salts
in the nutrient soil, and Pasteur suggested that in rabies, side by
side with a living microbe, there is possibly some chemical product
or anti-microbe which has, as in Raulin’s experiment, the power of
arresting the growth of the microbe. If we accept the theory of
arrest by some chemical product, we must suppose that in the
acquired immunity afforded by one attack of an infectious disease
this chemical substance is secreted, and, remaining in the system,
opposes the onset of the micro-organism at a future time. In the
natural immunity of certain species and individuals we must suppose
that this chemical substance is normally present. ,
Another theory is, that the micro-organisms assimilate the
elements which they require for their nutrition from the blood and
tissues, and render the soil impoverished or otherwise unsuitable for
54 BACTERIOLOGY.
the development of the same species of micro-organisms hereafter ;
this condition may be permanent, or the chemical constitution of
the tissues may be restored to normal, when immunity ceases. If,
however, we explain acquired immunity by the result of the growth
of a previous invasion of micro-organisms, we are still confronted ,
with the difficulty of explaining natural immunity.
A third theory is that the tissues are endowed with some
power of vital resistance to the development of micro-organisms,
similar to the vital resistance to coagulation of the blood, which is
supposed to exist in the-lining membrane of the healthy blood-
vessel; that in some species and individuals this exists to a high
degree, and hence their natural immunity. But this does not
explain how one attack confers immunity from a subsequent one,
One would expect that the vital resistance would invariably be
lowered by a previous attack, and increased liability be the constant
result,
A fourth theory was propounded by Metchnikoff, who maintains
that immunity depends upon phagocytosis. If anthrax bacilli are
inoculated in the frog, white blood-cells, or phagocytes, are observed
to incorporate and destroy them antil they entirely disappear, and
the animal is not affected. But if the animal, after inoculation,
is kept at a high temperature, the bacilli increase so rapidly that
they gain the upper hand over the phagocytes, and the animal
succumbs. ,
It has also been suggested that bacteria may attract or repel
the phagocytes, exercising either a positive or a negative chemio-
taxis, This power is supposed to depend upon some special product
of the bacteria. or possibly upon their toxins, as suggested by Roux.
We must suppose that the negative chemio-taxis has become changed
to a positive chemio-taxis in an immunised animal, so that the
phagocytes, instead of withdrawing and leaving the bacteria to
multiply, are readily drawn into the contest and destroy the
invaders, : .
In septicemia of mice, the white blood-cells are attacked and
disintegrated by the bacilli in a remarkable way. It is difficult,
however, to accept these observations as affording a complete ex-
planation of immunity. It is difficult to conceive that the leucocytes
in the blood and tissues in the field mouse are differently constituted
from those in the house mouse, so that they form an effectual
barrier to the onset of bacteria in the one case, though so readily
destroyed in the other, or that in acquired immunity the result is
due to educating the phagocytes to respond to a positive chemio-taxis.
IMMUNITY. 55
Phagocytosis cannot explain the immunity which results from
the injection of filtered cultures, or of antitoxins, but when
blood serum of immunised animals was shown to possess antitoxic
properties, a new explanation of immunity was at once forthcoming.
In the light of these djscoveries immunity, whether natural or
acquired, was regarded as due to the accumulation in the blood
and tissues of substances which have the property of counteracting
partially or entirely the products by which pathogenic bacteria
produce their poisonous effects, These antitoxins, or protecting
proteids, can be obtained not only from the blood but also from
the spleen and the lymphatic and other glands. They result
from the metabolism of the cells of the tissues of the body.
Phagocytes in their conflict with bacteria may play a small
part, but it is more than probable that immunity is altogether
independent of phagocytosis,
CHAPTER VI.
ANTITOXINS AND SERUM THERAPY.
Tr has been clearly shown by the experiments of Fodor and Nuttall
that some species of bacteria are killed by a mixture with fresh
blood. Fodor pointed this out in the case of the anthrax bacillus,
and Nuttall confirmed the experiments, and repeated them with
a number of different species of bacteria.
Behring and Nissen followed up this line of inquiry, and found
that there was a great difference in the behaviour of freshly drawn
blood to different bacteria. In some cases the bacteria were
destroyed, in others their growth was only retarded, and in others
again they were not affected at all. Bouchard pointed out that
although the normal blood serum of a rabbit may be used for the
cultivation of Bacillus pyocyaneus, the blood serum of a rabbit, which
has been rendered immune, will attenuate or entirely nullify the
pathogenic properties of the bacillus.
Ogata and Jasuhara obtained similar results by cultivating
anthrax bacilli in the blood of immune animals. Buchner demon-
strated that this property of fresh blood belonged to the serum
and not to the cellular elements, and strongly advocated the theory
that the force opposed to invading bacteria was to be found in the
serum rather than in phagocytes.
Similar experiments were made with the bacteria of swine-fever,
and Emmerich and Mastbaum discovered that the blood. serum of
immune rabbits could be used as a therapeutic agent to prevent
the progress of the disease in animals already showing symptoms
of infection.
A new light was thrown upon this question by the experiments
of Behring, Kitasato, Tizzoni and Cattani, and others in connection
with tetanus and diphtheria. In these diseases the bacteria do not
invade the body, but the poisonous principles elaborated at the
seat of inoculation are absorbed into the system and produce
deleterious effects. It was obvious that attention must be turned
towards counteracting or destroying these poisonous products.
56
ANTITOXINS AND SERUM THERAPY. 57
It was in this direction that the experiments of Behring and
Kitasato, in 1890, proved to be of profound importance. It was
shown that the blood serum of a rabbit rendered immune against
tetanus or diphtheria had no destructive or retarding effect on the
growth of the bacilli, but it possessed the power of neutralising the
poison developed by the agency of the bacilli. In short, the serum
was shown to possess an antitoxic instead of a bactericidal power.
Hankin conceived the idea that this property is due to substances
of the nature of defensive proteids, and the blood serum of the
naturally immune rat was found to contain a proteid body with well-
marked alkaline reaction, possessing the power of destroying anthrax
bacilli. Injection of this proteid into mice, together with fully
virulent anthrax spores, prevented the development of the disease.
Young rats are susceptible to anthrax, and, according to Hankin,
they can be protected from anthrax by injection of the blood serum
of the parent. Tizzoni and Cattani expressed the opinion that the
antitoxic substance in the blood serum of animals rendered immune
against tetanus is a globulin to which they gave the name tetanus
antitoxin. Buchner proposed the term alewins (dAcéw, I defend),
to signify these substances. Hankin subdivided them into sozins
and phylaxins. Sozins are defensive proteids occurring in normal
animals; phylaxins are only found in animals artificially immune ;
and each of these are sub-classed by Hankin according to their power
of attacking the bacteria themselves or the products they generate.
Myco-sozins :
Alkaline globulins from rat
(Hankin), destroying an-
pen ee sa,
Sozins: thrax bacillus.
Defensive proteids J
present in the nor-
: xX0-Sozins :
mal animal, To:
Of rabbit, destroying poison
of Vibrio Metchnikovi
\ (Gamaleia).
Defensive proteids
(Hankin) (Myco-phylaxins : : :
Alexins (Buchner) of rabbit, destroying pig
ins : typhoi acillus m-
Phylaxins : 2
Defensive _proteids merich).
present in the
animal after it has
artificially been
\ made immune.
4 Toxo-phylaxins :
Of rabbit, etc., destroying
diphtheria and tetanus
poisons (Behring and
Kitasato, anti-toxin of
\ Tizzoni and Cattani).
Tizzoni and Cattani immunised dogs and other animals against
tetanus, and employed the antitoxin as a therapeutic agent. Its
58 ... BACTERIOLOGY.
active substance was precipitated by alcohol. Behring, Kitasato, and
Schiitz experimented with a view to conferring immunity upon
horses. The cultures were mixed with terchloride of iodine, and
injected at intervals of eight days, and the antitoxic power tested
on mice. By using increasingly virulent cultures, the blood became
increasingly antitoxic..
Vaillard filtered tetanus cultures through porcelain, and heated
the filtrate at gradually diminishing temperatures. The first in-
jections were made with 10 cc., which had been raised to 60° C. for
an hour, then a filtrate was used which had been heated to 55° C.,
and lastly, a filtrate which had been heated to 50°C. The blood
became antitoxic, and by injecting increasing quantities of virulent
filtrates the antitoxic power was rapidly intensified, and animals
which were injected with antitoxin of full strength possessed
immunity many months afterwards,
Roux and Vaillard introduced another method. Virulent cultures
were filtered through porcelain, and the filtrate mixed with Gram’s
solution of iodine in iodide of potassium, To give immunity to a
rabbit, 3 cc. of toxin with 1 cc. of Gram’s solution were injected on
the first day, and increasing doses of toxin mixed with increasing
doses of Gram’s solution on the following days. The same method
was applied to horses, sheep, and cattle. The antitoxin was found
not only in the blood, but in the urine and saliva, and in the
milk in cows. With cows and goats it is necessary to proceed
with the utmost care; while horses, on the other hand, bear the
injections well, and are therefore more suitable for this purpose.
It is also very easy to obtain large quantities of blood from the
horse by inserting a trocar and cannula into the jugular vein.
Frankel was the first to produce immunity against diphtheria
by injecting guinea-pigs with toxin which had been heated to 70° C.
Behring mixed the toxin with terchloride of iodine, or employed small
doses of pure toxin. Horses, sheep, goats, and dogs were rendered
immune.
PREPARATION OF DIPHTHERIA ANTITOXIN.
For the preparation of diphtheria antitoxin Roux cultivates the
diphtheria bacillus in alkaline broth with 2 per cent. of peptone, and
by preference, in flasks in which the cultivating liquid can be exposed
to a current of moist air at 37°C. After about three weeks the
culture is filtered through a Chamberland filter, arid if tested on
a guinea-pig it will be found that jy of a cc. will kill an animal
weighing five hundred grammes in forty-eight hours. The diphtheria
ANTITOXINS AND.SERUM THERAPY. 59
toxin immediately before ‘the injection is mixed with 4 of its
volume of Gram’s solution. This is used for several weeks, and
afterwards only pure toxin is injected.
The horses employed for this purpose are animals no longer fit
for work, and it is necessary to inject them first of all with madlein
to be sure that they are not suffering from glanders.
In a horse inoculated by Roux, the injection began with 2 cc.
of iodised toxin, increased to 1 cc. by the thirteenth day, and the
injection continued daily. On the seventeenth day } cc. of pure
toxin was injected, and this was increased by the forty-first day
to 10 ce.; and on the forty-third day 30 cc, of pure toxin were
injected, causing pronounced edema. The doses were still further
increased, until on the eightieth day 250 cc. were injected. In
two months and twenty days the horse had received 800 ce. of
toxin.
On the eighty-seventh day the serum obtained had an immunising
power of over 50,000. By this is meant that a guinea-pig resisted
inoculation of 3 ce. of virulent diphtheria culture when injected
twelve hours beforehand with serum in quantity equal to the sggq5
part of its body weight. ‘
There are two tests which can be applied to the serum. First,
the antitoxic serum added to diphtheria toxin renders it inert; and,
secondly, if serum is injected into a guinea-pig and toxin injected
several hours afterwards, no result follows.
Several ways have been suggested for estimating the’ immunising
power of the serum.
In Ehrlich’s system, the unit is represented by °1 cc. of anti-
toxic serum, which, added to ‘8 cc. toxin, will neutralise it so that
the whole may be injected subcutaneously in a guinea-pig without
producing edema. The standard toxin is a toxin of which °3 ce.
is fatal to 1 kilo. of guinea-pig.
But the preventive power of the serum is best sepeee by the
result of a subsequent injection of toxin. The immunising power is
estimated by the number of grammes of guinea-pig which can be
protected against the minimum fatal dose of toxin by 1 ce. of anti-
toxic serum.
The antitoxic serum can be iten in sterilised flasks in the dark, .
with the addition of a small piece of camphor, or it may be dried
in vacuo, powdered, and thus supplied in a convenient form for trans-
port. It has merely to be dissolved in water before use.
Klein employed a modified plan by which he claimed to have
obtained antitoxin in a far shorter-time; than is possible by Roux’s
60 BACTERIOLOGY.
method. Unfiltered attenuated cultures were injected into the
horse. Later, large quantities of living diphtheria bacilli from
the surface of solid cultures, of gradually increasing virulence, were
repeatedly injected so as to allow the bacilli to grow and multiply.
In twenty-three days an antitoxic serum was obtained, one part
of which was found capable of protecting 20,000 to 40,000
grammes of guinea-pig against more than a fatal dose of both living
bacilli and the resulting toxin.
Serum Treatment of Diphtheria.—The results obtained by
Behring, Ehrlich, Kossel, and Wasserman, in the: treatment of
diphtheria in children in Germany by means of the curative serum,
and by Roux and others in France, led to the adoption of the treat-
ment in this country. It is best to use an especially constructed
hypodermic syringe, which can be easily taken to pieces, and placed
in boiling water to sterilise it. The skin surface of the flank is
washed, and disinfected with 1 in 20 carbolic, and the antitoxin is
then injected. The syringe is taken to pieces, placed again in boiling
water, and thoroughly cleaned,
The dose will depend upon the age of the patient and the
strength of the serum. From 10 cc. to 20 ce, are injected in children
under fifteen, and 30 cc. to 40 cc. in older patients, and the
injection may be repeated in 12 hours. The best results are said
to be obtained by injecting every 12 hours, for the first 12, 36 or
48 hours, according to the nature of the case, 1,000 Behring’s units,
this being the dose calculated according to the immunising power of
the serum. The result of the injection is to lower the temperature
and pulse, but frequently the reverse occurs, and in about half the
cases an urticarial and sometimes a scarlatiniform rash is produced.
Pains in the joints, in rare cases effusion, may also result from the
injection.
The beneficial results of the treatment are, according to the
Report of the Medical Superintendents of the hospitals of the
Metropolitan Asylums Board, as follows :—
(1) Diminution of the faucial swelling and of the consequent
distress ;
(2) Lessening or entire cessation of the irritating and offensive
discharge from the nose ;
(3) Limitation of the extension of membrane;
(4) Earlier separation of the exudation ;
(5) Limitation and earlier separation of membrane in laryngeal
cases ;
(6) Improvement in general condition and aspect of patients ;
ANTITOXINS AND SERUM THERAPY. 6]
(7) Prolongation of life, in cases which terminate fatally, to an
extent not obtained with former methods of treatment.
Statistics have also been brought forward which show, assuming
them to be reliable, a great reduction in the mortality after the
antitoxin treatment. A few instances may be quoted to illustrate
the statistical evidence.
According to Behring, in the four years prior to the employment
of antitoxin, there were in Berlin 15,958 cases of diphtheria, with a
mortality of 35:2 per cent. In 1894-5 there was an epidemic of
5,578 cases. Behring asserts that if the mortality had not been’
reduced by the antitoxin treatment 1,963 would have died instead
of 1,056. Behring also states that in the Charité Hospital there
were 299 patients, with 53 deaths, or 16-7 per cent, In the Bethania
Hospital, where antitoxin was not employed, there were 249
patients, with 112 deaths, or 43 per cent,
At Vienna, at the Anna Hospital for children, the mortality
in 760 cases was 50°65 per cent., but after the introduction of anti-
toxin there were 40 deaths in 159 cases, giving a mortality of
25:5 per cent.
In New York, it is said that before the introduction of anti-
toxin the mortality ranged from 30-67 to 37-34, while in 1895, under
treatment with antitoxin, the mortality fell to 19-43; but it was
also pointed out that since the introduction of antitoxin many
children with trifling attacks had been treated, and reported as
suffering from actual diphtheria, and that they would have recovered
without antitoxin, and therefore these cases have given the remedy
some credit which it does not deserve.
In London, according to the Report of the Medical Superin-
tendents of the hospitals of the Metropolitan Asylums Board there
were in 1894, before antitoxin was employed, 3,042 cases of
diphtheria with 902 deaths or 29°6 per cent., and in 1895, when
antitoxin was used, 3,529 cases with 796 deaths or 22°5 per cent. :
a reduction of 7-1 per cent, below that of 1894, The conclusions
drawn from the statistical and clinical observations are summed up
in the Report thus :—
The improved results in the diphtheria cases treated during the year
1895, are :-—
(1.) A great reduction in the mortality of cases brought under treat-
ment on the first and second day of illness.
(II.) The lowering of the combined general mortality to a point
below that of any former year.
(III.) The still more remarkable reduction in the mortality of the
laryngeal cases.
62 BACTERIOLOGY.
(IV.) The uniform improvement in the results of tracheotomy at
each separate hospital.
(V.) The beneficial effect produced on the clinical course of the
disease.
A consideration of the statistical tables and clinical observations,
covering a period of 12 months and embracing a large number of cases,
in our opinion sufficiently demonstrates the value of antitoxin in the
treatment of diphtheria.
It must be clearly understood, however, that to obtain the largest
measure of success with antitoxin it is essential that the patient be
brought under its influence at a comparatively early date—if possible not
later than the second day of disease. From this time onwards the chance
of a successful issue will diminish in proportion to the length of time
which has elapsed before treatment is commenced. This, though
doubtless true of other methods, is of still greater moment in the case
of treatment by antitoxin.
Certain secondary effects not infrequently arise as a direct result of
the injection of antitoxin in the form in, which it has at present to be
administered, and, even assuming that the incidence of the normal com-
plications of diphtheria is greater than can be accounted for by the
increased number of recoveries, we have no hesitation in expressing the
opinion that these drawbacks are insignificant when taken in conjunction
with the lessened fatality which has been associated with the use of this
remedy.
We are further of the opinion that in antitoxin serum we possess
a remedy of distinctly greater valne in the treatment of diphtheria than
any other with which we are acquainted.
On the other hand it has been urged that the decline in the
mortality in 1895 in London, which has been attributed entirely
to the antitoxin treatment, may possibly be partly due to the pre-
valence of a mild type of the disease, and that the fall in the
mortality during the seven previous years from 59 per cent. in
1888 to 29 per cent. in 1894, continued in 1895.
Tt is obvious that the whole subject requires to be very carefully
considered, and before any final conclusion can be arrived at as to
the therapeutic value of antitoxin, the evidence of others who have
had great experience in the treatment of diphtheria by the old and
the new methods must be taken into account, and reliable statistics
allowed to speak for themselves.
PREPARATION OF TETANUS ANTITOXIN,
Antitoxin for use in the serum treatment of tetanus is obtained
from the horse. The tetanus bacillus is cultivated in an atmosphere
of hydrogen, in flasks specially constructed for the purpose. In
ANTITOXINS AND SERUM THERAPY. 63
about a fortnight: the cultures are extremely toxic. The toxin is
obtained free from bacilli by filtration through porcelain. Injec-
tions may be given daily, subcutaneously or intravenously, beginning
with 1 ce, of iodised toxin, and gradually increasing the dose until
the pure toxin may be injected without danger.
- Roux and Vaillard produced immunity in about three months,
When a few days have elapsed after the last injection, the blood is
drawn, by means of a trocar and cannula, from the jugular vein into
a sterilised glass vessel, and set aside to coagulate ; next day the
serum is drawn off with a pipette, and used in the liquid state, or
dried in a vacuum over sulphuric acid, and subsequently powdered.
When required for use the powder is dissolved in cold water. About
5 grammes are used for a dose.
Serum Treatment of Tetanus.—The result, so far, of the
employment of tetanus antitoxin in animals suffering from tetanus
is disappointing, and the serum treatment is not likely to be of much
value in veterinary practice. Nocard infected sheep with tetanus
by inserting splinters of wood infected with spores into the muscles
of the leg. Tetanus supervened in eleven days, and the splinters
were removed, the tissues excised, and the wounds dressed with
iodoform. About twelve hours after the symptoms had shown
themselves, the sheep were inoculated with antitoxic serum at
intervals of one hour, but they all succumbed to tetanus. In one
case the total amount injected was 160 cc. of highly antitoxic serum.
The antitoxin has been employed in tetanus in man. Kanthack
has collected the history of a number of cases, and they indicate
that the treatment is useless in acute cases in man with a short
ineubation period, while chronic cases with a long incubation period
often recover after the treatment. At the same time it must be
remembered that recovery often took place in chronic cases before
the introduction of the antitoxin treatment.
The question must still be considered to be sub judice, and a
trustworthy conclusion can only be based upon a more extended use
-of the antitoxin and impartial reports of every individual case.
ANTITOXIN oF Septic INFECTIONS.
An anti-streptococcic serum has been prepared by Marmorek. A
culture of streptococcus was intensified in virulence by inoculation
from rabbit to rabbit, and highly virulent cultures gave rise to a
powerful toxin. Roget and Charrin also, found that the serum of
immunised rabbits and of a horse conferred immunity. A patient
64 BACTERIOLOGY,
with puerperal fever was injected with 8 cc., on the follow-
ing day with 16 cc., and on the third day with 25 cc. On the
fourth day the temperature had fallen, and the patient recovered.
Favourable results are said to have followed the use of the serum
in 46 cases of erysipelas,
Bokenham, working independently, cultivated the streptococcus
in a mixture of broth and serum. Horses and asses were inocu-
lated, and a considerable degree of immunity established. The
serum of an inoculated ass possessed antitoxic power.
Ruffer and Bullock succeeded in immunising four horses against
the toxin of Streptococcus pyogenes ; two had been previously immu-
nised against the toxin of the diphtheria bacillus. The streptococcus
was cultivated by Marmorek’s methods in a mixture of two parts
of blood-serum and one part of peptonised broth, and the virulence
of cultures maintained by inoculation of rabbits, On testing the
immunising power of the antitoxic serum on rabbits, the effect
appeared to be slight in comparison with the antitoxins of the
bacilli of diphtheria andtetanus. In treating cases of septic infection
in the human subject, it has been recommended to commence with
two injections of 10 cc., and.it is said that no unfavourable results
have been met with which could be attributed to the effect of the
serum.
Antiroxin oF TypHorp Fever anp OTHER DisEases.
An antitoxic serum has been obtained by Chantemesse for use in
cases of typhoid fever, and it is probable that attempts will be made
to extend the principle of the antitoxic treatment to other infective
diseases.
CHAPTER VII.
THE BACTERIOLOGICAL MICROSCOPE.
THE instruments sometimes in use in biological and pathological
laboratories are not sufficient for the study of bacteria. It is
absolutely essential for the examination of such minute objects that
the microscope should be equipped with an objective of sufficiently
high magnifying power and with a special illuminating apparatus,
while the mechanical arrangements of the stage must admit of the
examination of plate-cultivations. It would not be within the scope
of this work to give a detailed account of the mechanical arrange-
ments and optical principles of the microscope. These matters are
fully dealt with in special works on the subject,* but sufficient will
be said to afford assistance in the selection of a suitable instrument,
and to explain the improvements in the microscope which have been
such an aid in bacteriological investigations.
A magnified image of an object is the result of the change
produced in the direction of rays of light which are made to pass
through lenses. This alteration in the course of the rays is known
as refraction. A ray of light passing from a rarer into a denser
medium is refracted towards a line drawn perpendicularly to the
surface of the latter. A ray of light passing through air and
impinging on water will not pass on in the same direction, but will
be refracted towards a line drawn perpendicularly towards the
surface of the water. If the ray pass into glass instead of water
a greater refraction will take place, and if it pass into diamond the
bending in its course will be still greater (Fig. 11).
The sines of the angle of incidence and refraction of different
substances have a constant ratio to each other, which is known
as the index of refraction, and this is determined for different
substances by the refraction produced by the passage of rays from
avacuum. Thus the index of refraction for flint glass is about 1-6,
* Carpenter: Zhe Microscope, Nigeli and Schwenderer: The Microscope
in Theory and Practice.
65 5
66 BACTERIOLOGY.
the sine of the angle of incidence of a ray passing from a vacuum
into glass being to the sine of the index of refraction as 1:6 to 1.
If we study the course of a pencil of rays we find that some
of the rays are reflected instead of entering the medium and being
refracted. When, for example, a pencil of rays falls upon water
or glass, after passing through air, some of the rays are lost by
reflection, and the proportion of the lost rays will increase with
their obliquity. The diminution of the brightness of the image
when pencils of rays have to pass through lenses is thus accounted
for, and this loss of light increases when the number of surfaces
/Q /# ya
Fic. 11.—Tue Rerraction or Licut.
through which the rays pass are, as in high-power objectives,
increased. There is an additional loss when there is an increase in
the difference between the refractive power of the different media
through which light passes. When pencils of rays pass from glass
into air, and then into glass again, the loss is much greater than
when the air is replaced by a medium with a refractive index more
nearly approaching that of glass. This explains the value of the
immersion system, which will be referred to more fully later on,
and also the advantage of cementing pairs of lenses with Canada
balsam or glass paste. The lenses used in the optical arrangements
THE BACTERIOLOGICAL MICROSCOPE. 67
of a microscope are principally convex, and the imperfections which
result must, if possible, be entirely overcome. These imperfections
are spherical and chromatic aberration.
Spherical aberration results from the unequal refraction of
rays passing through lenses with equal curvatures. The rays passing
through an ordinary convex lens do not all come to the same focus.
The rays passing through the marginal portion come to a focus at a
7
R
2 1 Ua a
rid FoF
A |_|
l
R2
BR!
Fic. 12.—SpPHERIcAL ABERRATION.
point much nearer to the lens than the focus of the rays passing
through the more central portion of the lens (Fig. 12). If the whole
aperture of the lens is used there must of necessity be blurring, for
at the point at which the marginal rays form a distinct image the
central rays will be out of focus, and at the point at which the
central rays form a distinct image the marginal rays will have
diverged, causing indistinctness.
This is partially remedied by using a diaphragm
and shutting out the marginal rays; but this is [ee
at the cost of loss of light.and diminution of the
angle of aperture. The difficulty is approximately SH
overcome in practice by using a combination of
lenses. The aberration of a convex lens is the ‘ie.
opposite of that of a concave lens (Fig. 13). The -—*
makers of the best lenses endeavour to obtain this p. 13 Goy-
correction as perfect as possible to get the sharpness = giyattion oF
of the image, so essential in studying the mor- Lenses IN
phology of bacteria. Aspais Home
: . GENnEous Im -
Chromatic aberration is the result of the ji pasion.
unequal refrangibility of the coloured rays which
compose white light. If parallel rays of light pass through a
convex lens the violet rays, which are the most refrangible, will
come to a focus at a point much nearer to the lens than the
focus of the red rays, which are the least refrangible; and the
intermediate rays of the spectrum will be focussed at points between
the red and the violet. A screen held at either of these foci
will show an image with prismatic fringes (Fig. 14).
68 BACTERIOLOGY.
‘
The chromatic aberration may be reduced by stopping out the
marginal rays; but as it is necessary to get the most perfect
correction possible, advantage is taken of the different relations
which the refractive and dispersive powers bear to each other in
different glasses. By combining a double convex lens of crown
glass with a plano-convex lens of flint glass, correction is obtained
for the violet and red rays. An achromatic objective is constructed
on this principle, but the result is not perfect, as the intermediate
coloured rays remain uncorrected, and what is termed a secondary
spectrum gives rise to images with coloured fringes, especially at the
margin of the field. Abbé and Schott, after a great number of
experiments, succeeded in discovering a glass with optical properties
which removed the secondary spectrum, and objectives made with
the new glass are termed apo-chromatic. There is much more
Fic. 14.—CHRomatic ABERRATION.
perfect concentration of the component rays than in the ordinary
achromatic objectives, and the advantages thus obtained are very
great. The objectives can be made of higher angle and admit of
higher eye-pieces being used without materially diminishing the
brilliancy and definition of the image. There is a complete absence
of coloured fringes, and the perfect definition is invaluable in
micro-photography.
Another fault which has to be corrected is the aberration caused
by covering a microscopical preparation with a cover-glass. Ross
was the first to point out the difference in the image when the
object was examined under a cover-glass, and that by altering the
position of the front pair of lenses, in an objective corrected for an
uncovered object, the objective could be corrected for the covered
object (Fig. 15).
Objectives are generally corrected for a standard thickness of
cover-glass, but H. Lister devised a screw-collar adjustment by
which the position of the front pair of lenses could be altered at
will; and as it is almost impossible to obtain cover-glasses which
THE BACTERIOLOGICAL MICROSCOPE. 69
do not vary slightly in thickness, the most perfect definition
can only be obtained by adjusting for each separate cover-glass
preparation.
Immersion system.—All objectives were formerly used diry—
that is to say, with an air space between the objective and the
specimen to be examined—but high-power objectives are now almost
entirely made on the wnmersion system, a drop of liquid being
interposed between the objective and the cover-glass.
About fifty years ago Amici observed that if a drop of water
intervened between the cover-glass or an uncovered object and the
lens the image was more brilliant. The passage of rays from the
object or the cover-glass into air,
and again from air into glass, caused
considerable loss of light. With
objectives of wide angle of aperture
the advantages were counteracted
by the reflection of rays falling ob-
liquely upon the lens. By inter-
posing water more rays are bent
in or refracted, and enter the lens
instead of being reflected and lost.
Hartnack, Nachet, and others
adopted the immersion system, and
Fic. 15.—OBJective witH CoLLar
CorRECTION (0).
high-power water immersion lenses
were constructed with high angle
of aperture.* It was found that there was less necessity for
correcting for covers of different thickness, as the aberration from
this cause was diminished. The lenses were corrected for an average
thickness of cover, and slight deviations produced hardly any
appreciable effect.
Wenham, Stephenson, Abbé, and Zeiss carried the system to
perfection. They argued that the advantages obtained by water
immersion would be intensified if a liquid could be found of the
same refractive and dispersive power as crown glass. The media
would be optically uniform, and the result a homogeneous immersion
system.
* The angle of aperture is “the angle made by the most diverging of the
rays of the pencil issuing from any point of an object that can enter the lens,
and take part in the formation of an image of it.”
The numerical aperture is defined by Abbé as equal to “the sine of the
angle of aperture multiplied by the refractive index of the medium between
the object and the objective.”
70 BACTERIOLOGY.
After experimenting with different liquids—solutions of salts,
and various essential oils—Abbé recommended cedar oil as most
suitable for the purpose. In its optical properties it very closely
resembles crown glass, and it is far more convenient for use than
any watery solutions of salts, especially when it is necessary to
make a more or less prolonged examination of an object.
The difference between the dry, water, and oil immersion systems
may be illustrated, as Frinkel has pointed out, by a very simple
experiment. If a glass rod is inserted into an empty test-tube, it
is easily visible owing to the difference in refraction between the
glass and the surrounding air. If the tube is filled up with
water the rod is seen with difficulty, and if, instead of water, cedar
oil is used, the part of the rod immersed in the oil will entirely
disappear from view. The rays of light pass through an optically
uniform medium in the experiment with cedar oil, and no refraction
or reflection of rays of light can occur.
To use an oil immersion objective, a minute drop of cedar oil
is placed on the centre of the cover-glass, and the lens lowered
by means of the coarse adjustment until it touches the oil. The
specimen is then carefully brought into focus with the fine adjust-
ment. If the slide is held between the finger and thumb of one
hand, and moved from side to side while the other hand is working
the fine adjustment, there can be no danger of injuring either the
objective or the specimen.
Microscopes are made upon either the Ross or the Jackson
model. In the Ross model the body of the microscope is fixed
at its base to a transverse arm, which is raised or lowered with
it by the rack and pinion. In the Jackson model the body is
supported for a great part of its length on a solid * limb.”
In the Ross model, unless the body and transverse arm are very
solid as in Powell and Lealand’s microscopes (Fig. 23), there will be
vibration at the ocular end; but in the Jackson model vibration is
practically prevented, and this is most essential, especially in working
with very high powers.
The steadiness of the microscope also largely depends upon the
form of stand. There are four different types of stands. The
tripod (Fig. 23); the plate, with double columns; the single column,
ending in a plate or a bent claw; and the horse-shoe (Fig. 18).
The tripod stand with cork feet is the steadiest form of stand,
but it is cumbrous and expensive, and these objections also apply to
the model made by Ross.
The single upright should be unquestionably condemned, as it
THE BACTERIOLOGICAL MICROSCOPE. val
Fic. 16.—EneLish Mopet.
72 BACTERIOLOGY.
freely admits of vibration, and is most inconvenient for laboratory
work, The heavy horse-shoe form is compact and firm, and the
weight of it can hardly be considered an objection.
The tubular body is from eight to ten inches in length, and within
it is a draw-tube with engraved scale. By extending the draw-tube
greater magnification is obtained; but as this is at the cost of
definition it should hardly ever be used in the examination of
bacteria.
A triple nose-piece is a great convenience, saving the time which
is otherwise spent in replacing objectives of different magnifying
power, and there is less risk of injuring them.
Focus should be obtained by means of a rack and pinion coarse
Fic. 17.—RemovaBLlE MEcHANICAL STAGE.
adjustment. The sliding tube is not to be recommended, as the
motion may be stiff, encouraging the use of force, which in turn may
result in the objective being brought violently into contact with the
specimen, injuring the lens or damaging the preparation; or it may
get too loose and readily slip out of focus.
The stage should be flat and rigid, either rectangular or circular,
so long as it is sufficiently large to accommodate a plate-cultivation.
A removable mechanical stage is of great advantage for working
with high powers, as a motile bacterium can be constantly kept in
view while one hand is engaged in working the fine adjustment
(Fig. 17). It may also be employed as a finder if it is engraved
with a longitudinal and vertical scale, and provided with a stop.
The mechanical stage must be removable, so that the stage proper
THE BACTERIOLOGICAL MICROSCOPE.
Fig. 18.—ContinentaL Monet.
73
74 BACTERIOLOGY.
may be free from any attachments when required for the examina-
tion of cultures.
Diaphragms are necessary for regulating the amount of light.
The plan of using a series of discs, with apertures of different sizes,
should be avoided, as they are easily lost, and bacteriological investi-
gations may have to be made under conditions in which it is difficult
to replace them. A better plan is a revolving plate with apertures
of different sizes, but the most convenient form is the iris diaphragm
(Fig. 19).
The sub-stage condenser is quite as necessary in bacteriological
work as a high-power objective. In fact, the condenser and the
objective should be considered as forming one optical apparatus, and
the microscope regarded quite as.
incomplete without a condenser as
it would be without an objective.
By means of the sub-stage con-
denser (Fig. 20) the rays of light
are concentrated at one point or
on one particular bacterium ; and
for the best definition it is essential
that there should be mechanical
arrangements for accurately cen-
Fic. 19.—Iris DIAPHRAGM.
tring and focussing the condenser.
It may even with advantage be provided with a fine adjustment.
To sum up, a microscope for bacteriological investigation should
be provided with (1) a steady stand of either the tripod or horse-
shoe form; (2) a tubular body on the Jackson model; (3) a wide-
angled sub-stage condenser, such as Abbé’s; (4) objectives of an inch,
gth of an inch, and a j,th homogeneous immersion ; (5) a removable
mechanical stage ; and for the most accurate work there should be
centring arrangements and a coarse and fine adjustment to an oil-
immersion sub-stage condenser such as Powell and Lealand’s, and
a jth homogeneous oil-immersion apo-chromatic objective.
With regard to the choice of a microscope, it is chiefly a
question of price. The most perfect instrument is the large model
by Powell and Lealand, but it is most expensive, and quite unsuit-
able for laboratory work. For general use excellent instruments
are made by Zeiss, Leitz, Reichert, or Swift. The bacteriological
microscopes of these makers are in the necessary equipment
practically identical. The Zeiss microscope is the most finished, and
costs about twenty pounds. A similar microscope by Leitz and
by Swift costs about eighteen, and both make an excellent students’
THE BACTERIOLOGICAL MICROSCOPE. 75
bacteriological microscope, with a cheap form of adjustment to the
sub-stage condenser, at a total cost of about fifteen pounds.
Method of Illumination.—Good daylight is the best for
general work. The microscope should be placed near a window
with a northern aspect. Direct sunlight should never be utilised, and
the best light is that reflected from a white cloud. When daylight
is not available good results can be obtained with cither gas or a
Fic. 20.—ABBE’s CONDENSER CONSTRUCTED BY ZEISS.
paratin lamp. In the author’s laboratory the microscope lamps
are fitted with Welsbach incandescent mantles. These have many
advantages over an Argand burner or a paraffin lamp. A steady
and beautifully white light is obtained, and the lamps are quickly
lit, and require comparatively little attention. In using high powers
and carefully focussing the sub-stage condenser, the image of the
fabric of the mantle is embarrassing, and is an objection to this
light for the most accurate observations, but in other respects, and
76 BACTERIOLOGY.
for general use, it is the best form of artificial illumination for the
microscope.
An ordinary paraffin lamp of the cheapest form may be used,
but there are many objections to it, such as the shape of the
chimney, and the striz and defects in the glass. The best form of
paraffin lamp is constructed by Baker and by Swift from sug-
Fic. 21.—Microscore Lame.
gestions by Nelson and Dallinger (Fig. 21); there is also a similar
but much larger pattern which is made by Swift (Fig. 22). This
form of lamp has a large flat bowl for the oil. It is attached to
a standard, and can be raised or lowered to the desired position.
The chimney is of metal and blackened, so that there is no reflected
light, and it may also with advantage be provided with a shade,
so that no light reaches the eye except through the microscope.
The burner may be made to revolve, so that either the edge or
THE BACTERIOLOGICAL MICROSCOPE. 17
the flat of the flame may be utilised. Great care should be taken
to have the wick evenly trimmed. The best paraffin oil should
be burnt, and it is as well to add a small lump of camphor. The
metal chimney has an aperture in front, giving exit to the rays of
light, which is closed in by a slip of glass, The glass is very liable
to crack when exposed to the full force of the flame, and it is as well,
therefore, to be provided with a stock of glass slips, which have
‘isan
qt |
Fic. 22.,—Larce Microscope Lame.
been annealed by being enveloped in a cloth and boiled for two or
three hours.
The flat of the flame is used with low powers. The image of
the flame is reflected by a plane mirror, and a bull’s-eye condenser
interposed between the lamp and the mirror to give an equal
illumination of the whole field. In working with high powers the
lamp is turned with the flame edgewise, and the mirror is dispensed
with. By working, as it is termed, directly on the edge of the
flame, the illumination is greatly increased, and a band of light can
78 BACTERIOLOGY.
be concentrated on that part of the microscopical preparation which
requires most careful study (Fig. 23).
To obtain the best definition considerable time must be spent in
the arrangement of the illumination. The lamp and microscope
having been placed in position, a low power is first used and the
smallest diaphragm. On looking through the microscope it will
probably be observed that the image of the diaphragm is not in
the centre of the field. By moving the centring screw of the con-
denser this may be adjusted. The image of the edge of the flame
may not be central, and this must be adjusted by moving the lamp
into position. The low power is then replaced by a high power,
the largest diaphragm used, and the bacteria brought into focus,
The diaphragm must now be replaced by one of medium size, and
by racking the condenser up and down, a point will be arrived at
when the image of the edge of the flame appears as an intensely
bright band of light. If this is not exactly in the centre of the
field the centring screws of the condenser must again be adjusted.
Lastly, by trying different sizes of diaphragms, and focussing with
the fine adjustment, and using the correction collar, we arrive at the
sharpest possible image of the bacteria.
When the condenser has been accurately centred, it will still be
necessary to focus it for each individual specimen, so as to correct
for difference in the thickness of slides and the layers of mounting
medium. Correction for different thickness of cover-glasses must
in each case be made by means of the collar adjustment in the follow-
ing way. A high-power eye-piece is substituted for the ordinary
eye-piece, and the fault in the image will thereby be intensified. By
moving the collar completely round, first in one direction and then
the other, while carefully observing the effect on the image, it will
be seen to become obviously worse whichever way the collar is turned.
The collar must then be turned through gradually diminishing dis-
tances until an intermediate point is reached at which the best image
results with the high-power eye-piece, and on replacing this by the
low-power eye-piece the sharpest possible image will be obtained.
Effect of the sub-stage condenser.—The sub-stage condenser
gives the most powerful illumination when it has been racked up
until it almost touches the specimen. It produces a cone of rays of
very short focus, and the apex of the cone should correspond with the
particular bacterium or group of bacteria under observation. The
effect of the condenser without a diaphragm is to obliterate what
Koch has termed the structure picture. Tf the component parts of a
tissue section were colourless and of the same refractive power as
THE BACTERIOLOGICAL MICROSCOPE. 79
80 BACTERIOLOGY.
the medium in which the section is mounted, nothing would be
visible under the microscope. As, however, the cells and their nuclei,
and the tissue fibres do differ in this respect, the rays which pass
through them are diffracted, and an image of lines and shadows is
developed. If in such a tissue there were minute coloured objects,
and if it were possible to mount the tissue in a medium of exactly
the same refractive power, the tissue being then invisible, the
detection of the coloured objects would be much more easy. This
is exactly what is required in dealing with bacteria which have been
stained with aniline dyes, and the desired result can be obtained
by the'use of the sub-stage condenser.
Tf we use the full aperture of the condenser the greatly converged
rays play on the component parts of the tissue, light enters from
Fic. 24.—Ramspen MicroMETER HYE-PIECE.
all sides, the shadows disappear, and the structure picture is lost.
If now a diaphragm is inserted, so that we are practically only
dealing with parallel rays, the structure picture reappears. As the
diaphragm is gradually increased in size the structure picture
gradually becomes less and less distinct, while the colour picture,
the image of the stained bacteria, becomes more and more intense.
When, therefore, bacteria in the living condition and unstained tissues
are examined a diaphragm must be used, and when attention is
to be concentrated upon the stained bacteria in a section or in a
cover-glass preparation, the diaphragm must be removed and the
field flooded with light.
Micrometer.—For the measurement of bacteria a stage micro-
meter may be used with a camera lucida. The stage micrometer
consists of a slip of thin glass ruled with a scale consisting of tenths
and hundredths of a millimetre. The image of this can be projected
THE BACTERIOLOGICAL MICROSCOPE. 81
on a piece of paper, and a drawing made, and the object to be
measured can then be projected on the paper and compared with the
scale.
In the Ramsden micrometer eye-piece (Fig. 24) two fine wires
are stretched across the field of an eye-piece, one of which can be
moved by a micrometer screw. In the field there is also a scale
with teeth, and the interval between them corresponds to that of the
threads of the screw. The circumference of the brass head is usually
divided into one hundred parts, and a screw with one hundred threads
to the inch is used. The bacterium to be measured is brought into a
TN
Fic. 25.—MicromMEerer Eys-Precr By ZEISS.
position in which one edge appears to be in contact with the fixed
wire, and the micrometer screw is turned until the travelling wire
appears to be in contact with the other edge. The scale in the
field and the scale on the milled head together give the number of
complete turns of the screw and the value of a fraction of a turn in
separating the wires.
In the micrometer eye-piece constructed by Zeiss, the eye-piece
with a glass plate with crossed lines is carried across the field by
means of a micrometer screw (Fig. 25). Hach division on the edge
of the drum corresponds to ‘01 mm. Complete revolutions of the
drum are counted by means of a figured scale in the visual field.
Another method of measuring bacteria will be referred to in the
6
82 - BACTERIOLOGY.
chapter on micro-photography. The unit of measurement is one
thousandth of a millimetre or a micro-millimetre or micron, and is
expressed by the Greek letter y.
CARE OF THE MICROSCOPE.
After use the objectives, sub-stage condenser, and eye-piece
should be carefully wiped with soft linen, an old silk handkerchief,
or chamois leather, and the microscope covered with a bell-glass to.
protect it from dust. If a lens comes into contact with Canada
balsam it must be very carefully wiped with a soft rag moistened
with alcohol, and then cleaned with a soft leather. Microscopes
should not be exposed to the fumes of sulphuretted hydrogen,
chlorine, or volatile acids.
CHAPTER VIII.
MICROSCOPICAL EXAMINATION OF BACTERIA.
(4) Bacrerta in Liquips, Cutrurss, anD Fresu Tissuzs.
In conducting bacteriological researches the importance of absolute
cleanliness cannot be too strongly insisted upon. All instru-
ments, glass vessels, slides, and cover-glasses should be thoroughly
cleansed before use. A wide-mouthed glass jar should always be
close at hand, containing refuse aleohol for the reception of re-
jected slide preparations or dirty cover-glasses. When required
again for use, slides can be easily wiped clean with a soft rag. Cover-
glasses require further treatment, for, unless they are perfectly
clean, it is difficult to avoid the presence of air bubbles when
mounting specimens. They should be left in strong acid (hydro-
chloric, sulphuric, or nitric) for some hours; they are then washed,
first with water and then with alcohol, and carefully wiped with a
soft rag. The same principle applies in the preparation and
employment of culture media; any laxity in the processes of
sterilisation, or insufficient attention to minute technical details,
will surely be followed with disappointing results by contamination
of the cultures, resulting in the loss of much time.
For the preparation of microscopical specimens it will be found
convenient to use a platinum inoculating needle. This consists of
two or three inches of platinum wire fused into the end of a glass
rod about eight inches in length. Platinum is employed as it
rapidly cools after being raised to a white heat in the flame of a
Bunsen burner. It is thus completely sterilised, and in a few
moments is cool enough not to destroy the bacteria with which it is
brought into contact.
When using platinum needles, either for inoculating fresh tubes
in carrying on a series of pure cultures, or in transferring a small
portion of a cultivation to a cover-glass for examination under the
microscope, the careful sterilisation of the needle by heating the
83
84 BACTERIOLOGY.
platinum wire till it is white hot in every part, and heating also
as much of the glass rod as is made to enter the test-tube, must
be carried out with scrupulous care. Indeed it is a good plan to
Fic. 26.—InocuLtating NEEDLES.
let it become a force of habit to sterilise the needle before and after
use on every occasion, whatever may be the purposes for which it
is employed.
UnstatneD BactTERIA.
The bacteria in liquids, such as pus, blood, and culture-fluids, can
be investigated in the unstained condition by transferring a drop with
a looped platinum needle or a capillary pipette to a slide, covering
it with a clean cover-glass, and examining without further treat-
ment. If it is desirable to keep the specimen under prolonged
observation, a drop of sterilised water or salt solution must be run in
at the margin of the cover-glass to counteract the tendency to dry.
Cultures on solid media can be examined by transferring a small
portion with a sterilised needle to a drop of sterilised water on a
slide, thinning it out, and covering with a cover-glass as already
described.
Tissues in the fresh state may be teased out with needles in
sterilised salt solution, and pressed out into a sufficiently thin layer
between the slide and cover-glass. Glycerine may in many cases
be substituted for salt solution, especially for the examination of
micro-organisms such as Actinomyces and mould fungi.
There is, as a rule, no difficulty in recognising the larger micro-
organisms such as those just mentioned; but when we have to
deal with very small bacilli and micrococci, they may possibly be
mistaken for granular detritus or fat-crystals, or vice versa. They
are distinguished by the fact that fatty and albuminous granules
are altered or dispersed by acetic acid, and changed by solution
of potash; alcohol, chloroform, and ether dissolve out fat-crystals
MICROSCOPICAL EXAMINATION OF BACTERIA. 85
or fatty particles; on the other hand, micro-organisms remain
unaffected by these reagents. Baumgarten demonstrated tubercle
bacilli in sections by treating them with potash, which clarified
the tissues and brought the bacilli clearly into view. Actinomyces
and other vegetable structures will not disappear when sections are
immersed in weak hydrochloric acid and mounted in glycerine.
In examining unstained bacteria, it is necessary, in order to
obtain the structure picture, that the light entering the microscope
should be reduced by employing a small diaphragm, and the sub-stage
condenser carefully centred and focussed. To focus an unstained
specimen in which only bacteria are present, is often difficult. The
slide may be gently raised towards the objective, and the stage
may be constructed to enable this to be done with the index finger
(Fig. 16). If on tilting the slide the organisms come into focus it
will serve as a guide in working the fine adjustment. Another plan
when bacteria are examined in water, is to look for an air-bubble,
and then to focus its edge until the bacteria appear in view.
The simple method of covering the liquid with a cover-glass will
not answer for a prolonged examination, as the liquid evaporates and
the specimen dries up. To keep living bacteria under observation
for any length of time, in order to study their movements or spore-
formation, a special slide must be employed (p. 120).
SrainED BacTErRtia.
Weigert first pointed out the value of the aniline dyes for
staining bacteria, and we are principally indebted to Koch, Ehrlich,
Gram, and Léfier for many valuable processes.
The staining of fresh preparations, especially those with no
coagulable albumen to fix them, may be carried out by the method
of His. A slide is prepared as already described for the exami-
nation of micro-organisms in the fresh state. The reagents are
then applied by placing them with a pipette drop by drop at
one margin of the cover-glass, and causing them to flow through
the preparation by means of a strip of filter-paper placed at the
opposite margin.
Babés recommends another rapid means of examining cultivations.
A little of the growth, removed by means of a sterilised platinum
hook or small loop, is spread out on a cover-glass into as thin a
film as possible: when almost dry, a drop or two of a weak aqueous
solution of methyl violet is allowed to fall from a pipette upon the
film. The cover-glass with the drop of stain is, after a minute,
86 BACTERIOLOGY.
carefully turned over on to a slide, and the excess of stain gently
and gradually removed by pressure with a strip of filter-paper.
This affords a rapid means of demonstration—for example, of a
‘cultivation of Koch’s comma bacilli in nutrient gelatine—enabling
the microbes to be seen in some parts of the preparation both
stained and in active movement.
CovER-GLASS PREPARATIONS.
Bacteria may be spread out into a thin layer on a cover-glass,
and then treated with a dye, or sections of tissues containing bacteria
can be stained and then mounted in the usual way.
The method of making a cover-glass preparation is one which is
very commonly employed. In addition to its value as a means of
examining bacteria in liquids and solid culture media, it affords
the additional advantage of enabling, if necessary, a larve number
of preparations to be made, which, when dried, can be preserved,
stained or unstained, in ordinary cover-glass boxes; they are
then in a convenient form for transport, and can be mounted
permanently at leisure.
The method is as follows: A cover-glass is smeared with the
cut surface of an organ or pathological growth, or with sputum ;
or a drop of blood, pus, or culture-fluid is conveyed to it with a
looped platinum needle. It is absolutely necessary to spread out
the micro-organisms into a sutliciently thin layer, so that the
individual bacteria may be as much as possible in the same plane,
otherwise some in the field will be in focus and others out of
focus, and it would be impossible to obtain a satisfactory photograph
of such a specimen. To overcome this it will be necessary, in the
ease of cultures on solid media, to diffuse the bacteria in a little
sterilised water; and even cultures in liquids may sometimes with
advantage be diluted in the same way. By means of another
cover- glass the juice or fluid is squeezed out between them into a
thin layer, and on sliding them apart each cover-glass bears on one
side a thin film of the material to be examined; or a culture is
spread out into a thin film by means of a hooked platinum needle.
The cover-glass is then placed with the prepared side upwards, and
allowed to dry. After a few minutes, it is taken up with a pair of
flat-bladed or spring forceps, with the prepared side uppermost, and
passed rapidly from above downwards three times through the
flame of a spirit lamp or Bunsen burner, Two or three drops of
an aqueous solution of fuchsine or methyl violet will be sufficient to
cover the film, and after a minute or two the surplus stain is washed
MICROSCOPICAL EXAMINATION OF BACTERIA, 87
off with distilled water by means of a siphon apparatus or a wash-
bottle. The cover-glass may be allowed to dry, and then mounted
in Canada balsam, or it may, while still wet, be turned over on to a
slide, the excess of water removed with filter-paper, and the exposed
surface wiped dry. It may first be examined with a power of about
250 diams. ; and if a high magnification is required, which is usually
the case, a droplet of cedar oil is placed on the cover-glass, and the
specimen examined with an immersion lens.
If the specimen is to be made permanent, fix the cover-glass at
one corner with the thumb, and with a soft rag carefully wipe off
the cedar oil; then float off the cover-glass by running in distilled
water at its margin, and having made a little ledge with a strip of
filter-paper, place the cover-glass up against it upon one of its
edges and leave it to dry. When perfectly dry mount in Canada
balsam, or put it away in a cover-glass box provided with a label of
contents.
In many cases it is necessary or preferable to apply the stain
for a much longer period. This may best be effected by pouring
some of the staining solution into a watch-glass, and allowing the
cover-glasses to swim on the surface, with their prepared side, of
course, downwards. Throughout all these manipulations it is
necessary to bear in mind which is the prepared surface of the
cover-glass.
Instead of using the watery solutions of the aniline dyes the
author prefers in many cases to use stronger solutions, and to reduce
the staining by a momentary immersion in alcohol. Very beautiful
preparations of streptococci, sarcine and other bacteria can be
obtained by this method, which is as follows: Cover-glass prepara-
tions are stained with carbolised fuchsine (Neelsen’s solution) for
about two minutes, rinsed in alcohol for a few seconds, quickly
washed in water, and either examined in water or dried and
mounted in the usual way. The extent of decolorisation is a
matter of practice: a momentary immersion in alcohol is sometimes
sufficient ; too long immersion will remove too much of the colour ;
too short immersion will leave the delicate outlines indistinct. This
method is especially valuable for sarcine and streptococci, the
divisions between the elements being sharply defined, and as any
albuminous particles or débris in the preparation are decolorised,
much cleaner and sharper preparations are obtained than with the
watery solutions. lLéfler’s and other concentrated solutions may
also be used, but Neelsen’s solution may be regarded as the standard
one for this method.
88 BACTERIOLOGY.
Aniline oil, carbolic acid, and some other chemicals, when added
to the aniline dyes, have the property of acting in the manner
of mordants, in some way fixing the colour in the bacteria, so that
they are not so readily acted upon by decolorising agents.
Loffler’s Solution.—Potash intensifies the staining power, and
Koch and Léffler have both used it with methylene blue. Liffler’s
solution consists of 30 grammes of methylene blue in 100 grammes
of 1 in 10,000 solution of potash. It may be used with advantage
for almost all kinds of bacteria.
Gram’s Method.—With a solution of gentian-violet the whole
film on the cover-glass is at first stained violet. By immersing the
cover-glass in a solution of iodine in iodide of potassium the stain
is fixed in the bacilli, but not in any débris, pus cells, or tissue
elements present in the film. Consequently by transferring the
cover-glass to alcohol the bacilli alone remain stained, the violet
colour being merely changed to blue. By employing a contrast
colour, such as eosin, a double staining is obtained. In some
bacteria the sheath is by this method differentiated from the
protoplasmic contents.
The stock solution of gentian-violet is prepared by shaking up
1 ce. of pure aniline with twenty parts of distilled water, and.
filtering the emulsion.. Half a gramme of the best finely powdered
gentian-violet is dissolved in the clear filtrate, and the solution filtered
before use.
The details of the method will now be described. In the first
place, it is much better to employ the aniline-gentian-violet solution
quite freshly prepared, and the following useful method is invariably
used by the author: Place four or five drops of pure aniline in
a test-tube, fill it three-quarters full with distilled water, close the
mouth of the tube with the thumb, and shake it up thoroughly.
Filter the emulsion twice, and pour the filtrate into a watch-glass
or glass capsule. To the perfectly clear aniline water thus obtained
add drop by drop a concentrated alcoholic solution of gentian-violet
till precipitation commences. Cover-glasses must be left in this
solution about ten minutes, transferred to iodine-potassic-iodide
solution until in:two or three minutes the film becomes uniformly
brown, and then rinsed in alcohol. The process of decolorisation may
be hastened by dipping the cover-glass in clove-oil and returning it
again to alcohol. The cover-glass is once more immersed in clove-oil,
then dried by gently pressing between two layers of filter-paper,
and finally mounted in Canada balsam.
MICROSCOPICAL EXAMINATION OF BACTERIA. 89
DovusLe STAINING OF COVER-GLASS PREPARATIONS.
To double stain cover-glass preparations they can be treated by
Ehrlich’s method for staining tubercular sputum, or by Neelsen’s
modification, or by staining with eosin after treatment by the
method of Gram.
Ehrlich’s method is as follows: Five parts of aniline oil are
shaken up with one hundred parts of distilled water, and the
emulsion filtered through moistened filter-paper. A saturated
alcoholic solution of fuchsine, methyl-violet, or gentian-violet, is
added to the filtrate in a watch-glass, drop by drop, until precipitation
commences. Weigert recommended that exactly eleven parts of the
dye should be used to one hundred parts of the aniline solution.
Cover-glass preparations are floated in this mixture for fifteen
minutes to half an hour, then washed for a few seconds in dilute
nitric acid (one part nitric acid to two of water), and then rinsed
in distilled water. The stain is removed from everything except
the bacilli; but the ground substance can be after-stained brown
if the bacilli are violet, or blue if they have been stained red.
Neelsen’s Solution and Methylene Blue.—Ziehl suggested the use of
carbolic acid as a substitute for aniline oil, and Neelsen recommended
a solution composed of 100 cc. of a 5 per cent. watery solution
of carbolic acid, 10 cc. of absolute alcohol, and 1 gramme of
fuchsine. This stain is commonly known as the Neelsen or Ziehl-
Neelsen solution. Cover-glass preparations are floated on the hot
dye for two minutes, they are then rinsed in dilute sulphuric acid
25 per cent., washed in water, immersed in watery solution of
methylene blue for three minutes, again washed in water, dried,
and mounted in balsam.
Gram’s Solution and Hosin.—Double staining of cover-glasses can
be obtained by combining Gram’s method with eosin. The method
is very useful for differentiating the sheath of Streptococcus
pyogenes and Bacillus anthracis, from the protoplasmic contents,
and for staining preparations of pneumonic sputum, or of micrococci
and other micro-organisms in pus. After decolorising the prepara-
tion in alcohol, the cover-glass is transferred to a weak solution
of eosin for two or three minutes, then washed again in alcohol,
immersed in clove-oil, dried between filter-paper, and mounted in
balsam.
SraINING OF SPORES.
A slight modification of the ordinary process employed in making
cover-glass preparations has to be adopted to stain the spores of
90 BACTERIOLOGY.
bacilli. Under ordinary circumstances the stain will not penetrate
the sheath, but if it can be made to penetrate, it is not readily
removed. The cover-glass preparation must be heated to a tem-
perature of 210° C., for half an hour, or passed as many as twelve
times through the flame of a Bunsen burner, or exposed to the
action of strong sulphuric acid for several seconds, and then a few
drops of a watery solution, of an aniline dye may be applied in the
usual way.
To double stain spore-bearing bacilli the cover-glass preparations
may be floated, for from twenty minutes to an hour, on Ebrilich’s
fuchsine-aniline-water, or on the Ziehl-Neelsen solution. The stain
must be heated—by preference in a capsule placed in a sand-bath—
until steam rises. The fuchsine is removed from the bacilli by
rinsing in water and washing in weak hydrochloric acid, and then
the preparations are washed again in water, and floated for a few
minutes on a watery solution of methylene blue. They are again
rinsed in water, dried, and mounted. Neisser’s decolorising solution
consists of 25 parts of hydrochloric acid to 75 parts of alcohol.
STAINING OF FLAGELLA.
Koch first stained flagella by floating the cover-glasses on a
watery solution of hematoxylin. From this they were transferred
to a 5 per cent. solution of chromic acid, or to Miiller’s fluid, by
which the flagella obtain a brownish-black coloration. The author
succeeded in demonstrating and photographing flagella in prepara-
tions stained with a saturated solution of gentian violet in absolute
alcohol ; but these methods are now superseded owing to the much
more satisfactory method introduced by Loffler.
Léffler’s method depends upon the employment of a mordant.
Loffer tried tannate of iron, and after a number of experiments
the following method was introduced. An aqueous solution of
ferrous sulphate is added to an aqueous solution of tannin (20 per
cent.), until the mixture turns a violet-black colour, then 3 or 4 ce.
of a 1 in 8 aqueous solution of logwood are added. This constitutes
the mordant, and a few drops of carbolic acid may be added, and the
solution kept in well-stoppered bottles. The dye consists of 1 cc. of
a 1 per cent. solution of caustic-soda, added to 100 cc. of aniline
water, inwhich 4 or 5 grammes of either methyl violet, methylene blue,
or fuchsine, are dissolved. A cover-glass preparation is made in
the ordinary way, the bacteria being diffused in water, and then
spread out in a very thin film. After drying and very carefully
fixing, the film is covered with the mordant, and the cover-glass
MICROSCOPICAL EXAMINATION OF BACTERIA, 91
held over the flame until steam rises. The mordant is then washed
off with distilled water, and all traces removed from the edge of
the cover-glass with alcohol. The stain is filtered, and a few drops
allowed to fall on the film, and after a few minutes the cover-glass
is again very carefully warmed until steam rises. The stain is then
washed off with distilled water, and is ready to be examined and
subsequently mounted. For some bacteria it is necessary to modify
the solutions, either by the addition of acetic or sulphuric acid, or
by varying the quantity of soda solution.
Trenkmann introduced a modification of Léffler’s system. Cover-
glasses are floated for from two to twelve hours on a solution
consisting of 1 per cent. tannin and 2 per cent. hydrochloric acid.
After washing in water the preparation is stained with a saturated
aleoholic solution of any of the aniline dyes diluted in the propor-
tion of- 2 drops of the dye to 20 of water. The cover-glasses
which remain in the solution for from two to four hours are then
washed in water, and examined. The best results are obtained with
carbolised fuchsine, diluted in the proportion of 2 drops to 20 drops
of 1 per cent. carbolic. Trenkmann also recommended the use of
eatechu and logwood as mordants, with the addition of very dilute
acid, and subsequent staining with fuchsine.
Lutesch suggested the use of ferric acetate. To avoid any
deposit on the surface of the preparation, freshly prepared saturated
ferric acetate is used, and 5 to 10 drops of acetic acid are added to
16 cc. of the mordant. After warming the solution the preparation
is washed in water, followed by 20 per cent. acetic acid, again
thoroughly washed, and then stained with hot solution of fuchsine or
gentian-violet in aniline water.
Van Ermengem used a mordant composed of 1 part of
2 per cent. solution of osmic acid, 2 parts of 10 to 25 per cent.
solution of tannin, with to every 100 cc. of this mixture 4 or 5 drops
of acetic acid. A black ink is thus formed, and the solution is
applied for from five to thirty minutes. After washing in water and
alcohol the cover-glasses are placed in a solution of nitrate of silver
and transferred to another solution composed of 5 grammes of gallic
acid, 3 grammes of tannin, 10 grammes of acetate of soda, and 330
grammes of distilled water. In a few moments they are again placed
in nitrate of silver, and then washed and mounted in balsam.
Sclavo’s method answers well for certain micro-organisms. The
preparations are left for one minute in, solution of tannin, washed in
distilled water, transferred for a minute to 50 per cent. phospho-
molybdic acid, again washed and stained from three to five minutes
92 BACTERIOLCGY.
in hot saturated solution of fuchsine in aniline water, washed in water,
dried on filter paper, and mounted in balsam. The tannin solution
consists of 1 part of tannin to 100 cc. of 50 per cent. alcohol.
Nicolle and Morax also, have modified Loffler’s method. Per-
fectly clean cover-glasses are used, and the film is dried without
fixing in the flame. Cover-glasses are covered with the mordant,
and heated for about ten seconds, and when steam rises the mordant
is shaken off and the film rinsed with water. The same process is
repeated three or four times, and finally the cover-glass is stained
with Neelsen’s solution, holding it over the flame once or twice
for a quarter of a minute; it is then washed and examined.
Bunge prefers as a mordant a mixture of aqueous solution of
tannin with 1 in 20 aqueous solution of sesquichloride of iron in the
proportion of 3 parts of the tannin solution, 1 part of the iron
solution, with the addition of 1 ce. of a saturated watery solution of
fuchsine added to 10 ce. of the mixture. The mordant is kept before
use, and applied for five minutes. The preparation is then washed and
stained with Neelsen’s solution. In another plan the cover-glasses
are immersed for one half to one minute in 5 per cent. solution of
acetic acid, washed and dried. The mordant is then applied three or
four times, and the cover-glasses washed, dried, and then stained with
gentian-violet, dipped in 1 per cent. acetic acid, washed, dried, and
mounted. Peroxide of hydrogen may be added to the mordant,
drop by drop; it becomes reddish-brown in colour, and must be
shaken up and filtered before use. Cover-glasses are exposed to its
action for about a minute, and Neelsen’s solution is used for
staining.
Hessert dispenses with the mordant. The film is fixed by
treating cover-glasses with a saturated alcoholic solution of corrosive
sublimate. After washing, the cover-glass is stained for thirty to
forty minutes in a hot dye, by preference a 10 per cent, watery
solution of saturated alcoholic solution of fuchsine.
CovER-GLAss ImpREssIons.
One of the most instructive methods for examining micro-
organisms is to make an impression-preparation. This enables us,
in many cases, to study the relative position of individual micro-
organisms one to another in their growth on solid cultivating media,
and in some cases produces the most exquisite preparations for the
microscope. A perfectly clean, usually small-sized, cover-glass is
carefully deposited on a plate-cultivation, and gently and evenly
pressed down. One edge is then carefully levered up, with a needle,
MICROSCOPICAL EXAMINATION OF BACTERIA. 93
and the cover-glass lifted off by means of forceps. It is then
allowed to dry, passed through the flame three times, and stained
as already described. In some cases of plate-cultures, especially
where no liquefaction has taken place, the growth is bodily trans-
ferred to the cover-glass, and a vacant area left on the’ gelatine
or agar-agar, corresponding exactly with the form and size of the
cover-glass employed.
PRESERVATION OF PREPARATIONS.
After examining a cover-glass preparation with an oil immersion
objective the cedar oil must be carefully wiped off, and the slide
set aside for the Canada balsam to set. At a convenient time all
preparations should be sealed with a ring of Hollis’ glue; the
cedar oil used at subsequent examinations of the specimen will
not be able to work its way under the cover-glass, and prevent
the balsam from hardening. When it is ringed cedar oil can be
readily wiped off, and the specimen cleaned without danger of
moving the cover-glass and injuring the preparation.
(B) Bacrerra 1x Sections or Tissues.
Methods of Hardening and Decalcifying Tisswes.—To harden small
organs, such as the viscera of a mouse, they should be placed on
a piece of filter-paper at the bottom of a small wide-mouthed glass
jar, and covered with about twenty times their volume of absolute
alcohol. Larger organs, pathological growths, etc., are treated in
the same way, but must first be cut into small pieces, or cubes,
varying from a quarter of an inch to an inch in size. Miiller’s
fluid may also be employed, and methylated spirit may be sub-
stituted for alcohol, from motives of economy. Tissues hardened in
absolute alcohol are ready for cutting in two or three days, and
those hardened in Miiller’s fluid in as many weeks.
Teeth, or osseous structures, must first be placed in a decalcifying
solution, such as Kleinenberg’s. When sufficiently softened, they
are allowed to soak in water, to wash out the picric acid, and then
transferred through weak spirit to absolute alcohol. Ebner’s solu-
tion also gives excellent results, especially when the structures to
be decalcified are placed in fresh solution from time to time.
Methods of Embedding, Fixing, and Cutting.—The author
finds that freezing with ether combined with the method of em-
bedding in celloidin gives excellent results. The pieces of tissue
to be embedded are placed, after the process of hardening is com-
94 BACTERIOLOGY.
pleted, in a mixture of ether and alcohol for an hour or more.
They are then transferred to a solution of celloidin in equal parts
of ether and alcohol, and left there, usually for several hours.
The piece of tissue is then placed in a glass capsule, and some
of the celloidin solution poured over it. The capsule can be placed
Fic. 27.—Swirt’s Freezing MIcRoTome.
bodily in 60 to 80 per cent. alcohol, and left until the following
morning. The celloidin will then be of the consistency of wax.
The piece of tissue is next cut out, and after trimming off superfluous
celloidin is put in water until it sinks. It is then transferred to
gum, and frozen and cut with a freezing microtome.
For cutting with Jung’s microtome, the tissues are embedded
MICROSCOPICAL EXAMINATION OF BACTERIA. 95
in paraffine or celloidin, and mounted on cork ; or, if firm enough,
they may be fixed upon cork without any embedding material at
all. Paraffine, dissolved in chloroform, will be found very service-
able as an embedding material.
Corks ready cut for the clamp of the microtome are smeared
over with the solution of celloidin. This can be applied with a
glass rod to the surface which is to receive the piece of tissue.
The corks are then set aside for the film of celloidin to harden.
In the case of lung, or degenerated broken-down tissue, the
specimen should be left for a much longer time than is found to
be sufficient for firmer structures. When ready, it is removed
from the celloidin solution with forceps and placed upon the pre-
Fic. 28.—June’s MicroTome.
pared cork. Enough of the solution, which is of syrupy consistence,
is allowed to fall on the piece of tissue to cover it completely, and
the mounted specimen is placed in the alcohol to harden. The
specimen will be ready for cutting next day.
The specimen may be more neatly embedded by fixing it with
a pin in a small paper tray, pouring the celloidin solution over it,
and then placing the tray in alcohol to harden the celloidin. The
embedded specimen is then fixed on a cork, which has been cut for
the clamp of the microtome. The celloidin in the section disappears
in the process of clearing with clove-oil.
In the case of specimens embedded in celloidin, or mounted
directly on a cork, the tissue, as well as the blade of the knife, should
be kept constantly bathed with alcohol, and the sections transferred
from the blade with a camel’s-hair brush, and floated in alcohol.
96 BACTERIOLOGY.
For fixing directly on cork, small organs and pieces of firm tissue
such as the kidneys of a mouse, or liver, we may employ gelatine or
glycerine gelatine, liquefied over a Bunsen burner in a porcelain
capsule. Glycerine gelatine may be used with advantage for fixing
irregular pieces of tissue, as it does not become of a consistency
that would injure the edge of the knife. The cork, with specimen
affixed, is placed in alcohol, and is ready for cutting sections next
day.
Material infiltrated with paraffine must be cut perfectly dry,
‘and the sections prevented from rolling up by gentle manipulation
witb a camel’s-hair brush. They must then be picked off the blade
of the knife with a clean needle, and dropped into a watch-glass
containing xylol. This dissolves out the paraffine. The sections are
then transferred to alcohol to get rid of the xylol, and then to the
staining solution.
Staining Bacteria in Tissue Sections.—Sections of fresh tissues
made with the freezing microtome are to be floated in ‘8 per cent.
salt solution, and then carefully transferred, well spread out on a
platinum lifter, to a watch-glass containing absolute alcohol. Simi-
larly, sections selected from those cut with Jung’s microtome may
be transferred from the spirit to absolute alcohol. The sections
may be then stained by any of the methods to be described.
It is often advisable to employ some method which will enable
one to study the structure of the tissue itself; and sections, however
stained, should always be first examined with low powers, to enable
one to recognise the tissue under examination, and to examine in
many cases the topographical distribution of masses of bacteria.
With a power of about 250 diams. (one-sixth), very many bacteria
can be distinguished ; and with the oil immersion lenses the minutest
bacilli and micrococci can be recognised, and the exact form of
individual bacteria accurately determined. As most good modern
instruments are provided with a triple nose-piece, there is no loss
of time in examining a preparation successively with these different
powers.
Weigert’s Method.—A very useful method for staining both
the tissue and the bacteria is as follows: Place the sections for
from six to eighteen hours in a 1 per cent. watery solution of any of
the basic aniline dyes (methyl violet, gentian violet, fuchsine, Bis-
marck brown). To hasten the process, place the capsule containing
the solution in the incubator, or heat it to 45° GC. A stronger
solution may also be employed, in which case the sections are far
more rapidly stained, and are easily over-stained. In the latter case
MICROSCOPICAL EXAMINATION OF BACTERIA. 97
they must be treated with a half-saturated solution of carbonate of
potash. In either case the sections are next washed with distilled
water, and passed through 60 per cent. alcohol into absolute alcohol.
When almost decolorised, spread out the section carefully on a
platinum lifter and transfer it to clove-oil, or stain with picro-carmine
solution (Weigert’s) for half an hour, wash in water, alcohol, and
then treat with clove-oil. After the final treatment with clove-oil,
transfer with the platinum lifter to a clean glass slide. Dry the
preparation by pressure with a piece of filter-paper folded several
times, and preserve in Canada balsam, dissolved in xylol.
Gram’s Method.—In the method of Gram sections are stained
for ten minutes in a capsule containing aniline-gentian-violet solution.
Great care must be taken not to injure the sections. If there is
any difficulty in finding them, it is best to carefully pour off the
stain and fill up the capsule with water. The sections are then readily
visible, and can be taken up on the end of a glass rod and placed
in the iodine and iodide of potassium solution, where they remain for
two or three minutes, until stained uniformly brown and resembling
in appearance a boiled tea-leaf. They are then placed in absolute
alcohol, and washed by carefully moving the sections in the liquid
with a glass rod. When completely decolorised they are spread out
on a lifter, and transferred to clove-oil until completely clarified.
Each is transferred with a lifter to a slide, and the clove-oil is
run off and then completely removed by gently pressing two or
three layers of filter-paper upon the section. Finally, the section
is mounted in Canada balsam.
The process of decolorisation may be hastened by transferring the
section from alcohol to elove-oil, and back again to alcohol, repeating
this two or three times.
On examination the tissue appears colourless, or slightly tinged
yellow from too long immersion in the iodine solution, while the
micro-organisms are stained blue or blue-black.
Double staining is obtained by transferring the sections after
decolorisation to eosin, Bismarck brown, or vesuvin. They are left
in a watery solution for two or three minutes, then again washed in
alcohol, before clarifying in clove-oil and mounting in balsam.
Another instructive method is to place the decolorised sections
in picro-carminate of ammonia for three or four minutes, and then
treat with alcohol and clove-oil.
A similar result is obtained by placing the sections in Orth’s
solution (picro-lithium carmine), transferring to acidulated alcohol,
and then passing through clove-oil and mounting in balsam.
7
98 BACTERIOLOGY.
In Ehrlich’s method delicate sections are liable to be injured by
immersion in the nitric acid, and therefore Watson-Cheyne suggested
the use of formic acid.
The Ziehl-Neelsen method, in which sulphuric acid is used instead
of nitric acid, is much to be preferred to Ehrlich’s method.
Ziehl-Neelsen Method:—The solution is warmed, and sections
left in it for ten minutes. The red colour, which disappears when
the section is placed in weak sulphuric acid (25 per cent.), may
partly return when the section is placed in water. In this case the
section must be again immersed in acid and passed backwards and
forwards from acid to water until the red colour has completely, or
almost completely, disappeared. It must be thoroughly washed in
water to remove all traces of the acid, and then placed in a watery
solution of methylene blue for two or three minutes, washed again
in water, immersed in alcohol, clarified in clove-oil, and mounted in
the usual way. Sections are brilliantly stained, and the results are
very permanent.
Many special methods of staining have been introduced, and will
be given in subsequent chapters with the description of the bacteria
to which they apply. The methods already described are those
which are more or less in constant use in studying bacteria and in
conducting original researches.
CHAPTER IX.
PREPARATION OF NUTRIENT MEDIA AND METHODS OF
CULTIVATION.
To cultivate micro-organisms artificially, and, in the case of the
pathogenic bacteria, to fulfil the second of Koch’s postulates, they
must be supplied with nutrient material free from pre-existing
micro-organisms. Hitherto various kinds of nutrient liquids have
been employed, and in many cases they still continue to be
used with advantage, but for general use they have been, in a
great measure, supplanted by the methods of cultivation on sterile
solid media about to be described. The advantages of the latter
methods are numerous. In the first place, in the case of liquid
media, in spite of elaborate precautions and the expenditure of much
labour and time, it was almost impossible or extremely difficult to
obtain a pure culture. When a drop of liquid containing several kinds
of bacteria is introduced into a liquid medium, we have a mixed
cultivation from the very first. If in the struggle for existence
some bacteria were unable to develop in the presence of others, or
a change of temperature and soil allowed one form to predominate
over another, then we might be led to the conclusion that many
bacteria were but developmental forms of one and the same micro-
organism ; while possibly the contamination of such cultures might
lead to the belief in the transformation of a harmless into a patho-
genic bacterium. The secret of the success of Koch’s methods greatly
depends upon the possibility, in the case of starting with a mixture
of micro-organisms, of being able to isolate them completely one
from another, and to obtain an absolutely pure growth of each
cultivable species. When sterile nutrient gelatine has been liquefied
in a tube and inoculated with a mixture of bacteria in such a way
that the individual micro-organisms are distributed throughout it,
and the liquid is poured out on a plate of glass and allowed to solidify,
the individual bacteria, instead of moving about freely as in a liquid
medium, are fixed in one spot, where they develop individuals of
99
100 BACTERIOLOGY’.
their own species. In this way colonies are formed each possessing
its own biological characteristics and morphological appearances.
When an adventitious germ from the air falls upon the culture, it also
grows exactly upon the spot upon which it fell, and can be easily
recognised as a stranger. To maintain the individuals isolated from
one another during their growth, and free from contamination, it is
only necessary to thin out the cultivation, and to protect the plates
from the air. The slower growth of the micro-organisms in solid
media, affording much greater facility for examining them at various
intervals and stages of development, is an additional point in favour
of these methods; and the characteristic macroscopical appearances
so frequently assumed are, more especially in the case of morpho-
logical resemblance or identity, of the greatest importance. ‘The
colonies on nutrient gelatine (examined with a low power) of micro-
organisms such as Bacillus anthracis and Proteus mirabilis, the
naked eye appearances in test-tubes of the growth of the bacilli
of anthrax and tubercle, and the brilliant growth of Micrococcus
prodigiosus, may be quoted as examples in which the appearances are
often very striking and sometimes quite characteristic.
Sorip Menta.
i
(A) Preparation or Nutrient GELATINE AND NUTRIENT
AGAR-AGAR.
Nutrient Gelatine is prepared as follows: Take half a kilo-
gramme of beef (one pound), as free as possible from fat. Chop it
up finely, transfer it to a flask or cylindrical
vessel, and shake it up well with a litre of
distilled water. Place the vessel in an ice-
pail, ice-cupboard, or in winter in a cold
cellar, and leave for the night. Next morn-
ing commence with the preparation of all
requisite apparatus. Thoroughly wash and
rinse with alcohol about 100 test-tubes, and
allow them to dry. Plug the mouths of the
test-tubes with cotton-wool, taking care that
the plugs fit firmly but not too tightly.
Place them in their wire cages in the hot-air
steriliser, to be heated for an hour at a temperature of 150° C. In
the same manner cleanse and sterilise several flasks and a small
glass funnel. In the meantime the meat infusion must be again
Fic. 29.—Wire Cace
FOR TEST-TUBES.
well shaken, and the liquid portion separated by filtering and
DESCRIPTION OF PLATE II.
Pure-cultivations of Bacteria.
Fig. 1.—ln the depth of Nutrient Gelatine. A pure-cultivation of Kochs
comma-bacillus (Spirillum cholerze Asiatic) showing in the track of
the needle a funnel-shaped area of liquefaction enclosing an air-bubble,
and a white thread. Similar appearances are produced in cultivations of
the comma-bacillus of Metchnikoff.
Fig. 2.—On the surface of Nutrient Gelatine. A pure-cultivation of Bacillus
typhosus on the surface of obliquely solidified nutrient gelatine.
Fie. 3.—On the surface of Nutrient Agar-agar. Pure-cultivation of Bacillus
indicus on the surface of obliquely solidified nutrient agar-agar. The
growth has the colour of red sealing-wax, and a peculiar crinkled
appearance. After some days it loses its bright colour and becomes
purplish, like an old cultivation of Micrococcus prodigiosus.
‘FIG. 4.—On the surface of Nutrient Agar-agar. A pure-cultivation obtained
from an abscess (Staphylococcus pyogenes aureus).
Fig. 5.—On the surface of Nutrient Agar-agar. A pure-cultivation obtained
from green pus (Bacillus pyocyaneus). The growth forms a whitish,
transparent layer, composed of slender bacilli, and the green pigment
is diffused throughout the nutrient jelly. The growth appears green by
transmitted light, owing to the colour of the jelly behind it.
Fie. 6.—-On the surface of Potato. A pure-cultivation of the bacillus of
glanders on the surface of sterilised potato.
Plate IL
Fig 6
Fig 5
Fig 4
Fig3
Fig 1.
Vincent Brocks,Day & Son, Lith
akshanle,fecit
ATIONS.
Se
V
PURE-CULTI
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 101
squeezing through a linen cloth or a meat press. The red juice thus
obtained must be brought up to a litre by transferring it to a large
measuring glass and adding distilled water. It is then poured
into a sufficiently large and strong beaker, and set aside after the
addition of 10 grammes of peptone, 5 grammes of common salt and
100 grammes of best gelatine.
In about half an hour the gelatine is sufficiently softened, and
subsequent heating m a water-bath causes it to be completely
Fanaa _
|
Fic. 30.—Hor Arr STeRiLiser.
dissolved. The danger of breaking the beaker may be avoided by
placing a cloth, several times folded, at the bottom of the water-bath.
The next process requires the greatest care and attention. Some
micro-organisms grow best in a slightly acid, others in a neutral
or slightly alkaline, medium. For example, for the growth and
characteristic appearances of the comma bacillus of Asiatic cholera
a faintly alkaline soil is absolutely essential. This slightly alkaline
medium will be found to answer best for most micro-organisms, and
may be obtained as follows :—-
With a clean glass rod dipped in the mixture, the reaction
upon litmus-paper may be ascertained, and a concentrated solution
of carbonate of soda must be added drop by drop, until red litmus-
102 BACTERIOLOGY.
paper becomes faintly blue. If it has been made too alkaline, it can
be neutralised by the addition of lactic acid.
Finally, the mixture is heated for an hour in the water-bath.
Ten minutes before the boiling is completed, the white of an egg
beaten up with the shell is added, and the liquid is then filtered
while hot. For the filtration, the hot-water apparatus (Fig. 31)
can be used with advantage, furnished with a filter of Swedish
paper, which may be conveniently
i} made in the following way :—
i About eighteen inches square
| of the best and stoutest filter paper
egugTTO rrp is first folded in the middle, and
Fy then creased into sixteen folds. The
| Gaull i Will =
:S filter is made to fit the glass funnel
O « by gathering up the folds like a fan,
~ and cutting off the superfluous part.
The creasing of each fold should be
made firmly to within half an inch
of the apex of the filter, which part
is to ke gently inserted into the
tH tube of the funnel. To avoid
la bursting the filter at the point, the
i)
<i ir Vim
{|
HI
broth, when poured out from the
|
i
Hi flask, should be directed against
Mt the side of the filter with a glass
i
lt UO)
=a Ze
rod. During filtration the funnel
== should be covered over with a
= circular plate of glass, and the pro-
cess of filtration must be repeated,
Fic. 31.—Hor-water Firrerrng if necessary, until a pale, straw-
APPARATUS. coloured, perfectly transparent
filtrate results.
The sterilised test-tubes are filled to about a third of their depth
by pouring in the gelatine carefully and steadily, or by employing a
small sterilised glass funnel. The object of this care is to prevent
the mixture touching the part of the tube with which the plug
comes into contact ; otherwise, when the gelatine sets, the cotton-
wool adheres to the tube and becomes a source of embarrassment in
subsequent procedures. As the tubes are filled they are placed in
the test-tube basket, and must then be sterilised. They are either
lowered into the steam steriliser, when the thermometer indicates
100° C., for twelve minutes for four or five successive days, or they
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 103
may be transferred to the test-tube water-bath, and heated for an
hour a day for three successive days.
Tf the gelatine shows any turbidity after these processes it must
Fic. 32.—MeEtTHOD OF MAKING A FOLDED Fitter.
be poured back from the test-tubes into a flask, boiled up for ten
minutes, and filtered once more, and the processes of sterilisation just
described must be repeated.
Fic. 33.—STEAM STERILISER.
Nutrient Agar-agar.—Agar-agar is a substance prepared
from seaweed which grows on the coasts of Japan and India,
and is supplied in long crinkled strips. It boils at 90° C., and
104 BACTERIOLOGY.
remains solid up to a temperature of about 45° C. It is there-
fore substituted for gelatine in the preparation of a jelly for the
cultivation of those bacteria which will only grow, or grow best, in
the incubator at the temperature of the blood. It may also be
employed at ordinary temperatures for bacteria which liquefy
gelatine. The preparation is conducted on much the same principles
as those already described. Instead, however, of 100 grammes of
gelatine, only about 20 grammes of agar-agar are employed (1:5 to
2 per cent.), and to facilitate its solution it must be allowed to soak
in salt water overnight. For the filtration, flannel is substituted
for filter-paper, or may
be used in combination
with the latter. The
hot-water apparatus
is invariably employed,
unless, to accelerate
the process, the glass
funnel and receiver are
bodily transferred to
the steam steriliser. If
the conical cap cannot
be replaced, cloths laid
over the mouth of the
steriliser must be em-
ployedinstead. It may
be necessary to repeat
the process of filtra-
tion, but it must not be
expected that such a
briliant transparency
Fic. 34.—Incupator. can be obtained as with
gelatine. The final
result, when solid, should be colourless and clear; but if slightly
milky, it may still be employed.
A little liquid gradually collects in the tubes, being expressed by
the contraction of the agar-agar.
Wort-gelatine is used in studying the bacteria of fermentation.
It is made by adding from 5 to 10 per cent. of gelatine to beer-wort.
Glycerine Agar-agar.—This is prepared by adding 5 per
cent. of glycerine to nutrient agar-agar, after the boiling and before
the filtration, and other modifications can be made for special
purposes by the addition of grape-sugar or of gelatine.
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 105
After the final treatment in the steam steriliser some of the
tubes of gelatine and agar-agar are placed upright and allowed to
set, and others are placed on an inclined plane or in the blood-serum
inspissator, and left to gelatinise with an oblique surface.
(8) Mernops of empLoyinc Nurrient Jeniy 1n Trst-TuBEs
AND oN GLAss PLa‘vEs.
Test-tube-cultivations.—To inoculate test-tubes containing
nutrient jelly, the cotton-wool plug is first twisted round in case
there are any adhesions between the plug and the test-tube. It is
then removed with the thumb and
index finger of the right hand, and q
placed between the fourth and fifth |
fingers of the left hand, instead of
being put down on the laboratory table
and thereby probably contaminated
with bacteria or the spores of mould
fungi. A sterilised needle charged,
for example, with blood or pus con-
taining bacteria, or with a colony from
a plate-culture, is thrust once in the
middle line into the nutrient jelly,
and steadily withdrawn. The tube
should be held horizontally or with
its mouth downward, to avoid, as far
as possible, accidental contamination
from the gravitation of germs in the
Fic. 35.—Metuop or [nocunat-
; ; ING A TEST-TUBE CONTAINING
as possible. ‘The cotton-wool project- Qppring Nurrient JELLY.
ing beyond the mouth of the tube is
then thoroughly burnt in the flame of a Bunsen burner or blow-
air; and the plug replaced as quickly
pipe, and an india-rubber cap fitted over the mouth of the tube
The chances of error arising from contamination of the culti-
vations are reduced by avoiding draughts at the time of inoculation,
and it is best that these manipulations should be carried on in a
quiet room in which the tables and floor are wiped with damp cloths,
rather than in a laboratory in which the air becomes charged with
germs through constant sweeping and dusting, and the entrance
and exit of classes of students. In conducting any investigation
a dozen or more tubes should be inoculated, and if by chance an
adventitious germ, in spite of all precautions, gains an entrance,
106 BACTERIOLOGY.
the contaminated tube can be rejected, and the experiments con-
tinued with the remaining pure cultivations.
When, however, one tube containing a liquid medium is in-
oculated from another, as in the process of preparing plate-cultures,
or when a culture is made from a tube in which the growth has
liquefied the gelatine, it is obvious that the tubes cannot be inverted
or held horizontally, and they must then be held and inoculated as
in Fig. 38. To inoculate those tubes of nutrient media which have
been solidified obliquely, the point of a straight sterilised needle
charged with the material to be cultivated is traced over the surface
of the jelly from below upwards, or the inoculated material may be
spread out with a hooked or looped needle.
Examination of Test-tube-cultivations—The appearances pro-
duced by the growths in test-tubes can be in most cases sufficiently
examined with the naked eye. In some cases the jelly is partially
or completely liquefied, while in others it remains solid. The
growths may be abundant or scanty, coloured or colourless. The
nutrient jelly may itself be tinged or stained with products resulting
from the growth of the organisms. When liquefaction slowly takes
place in the needle track, or the organism grows without producing
this change, the appearances which result are often very delicate,
and in some cases very characteristic. The appearance of a simple
white thread, of a central thread with branching lateral filaments,
of a cloudiness, or of a string of beads in the track of the needle,
may be given as examples.
In some cases much may be learnt by examining the growth with
a magnifying glass. Here, however, a difficulty may be encountered,
for the cylindrical form of the tube so distorts the appearance of its
contents, that the examination is rendered somewhat difficult. To
obviate this, a very simple contrivance may be employed with
advantage. This consists of a rectangular vessel, about four inches
in height and two inches in width, which may be easily constructed
by cementing together two slips of glass to form the back and front,
with three slips of stout glass with ground edges forming the sides
and base (Cheshire). The front may be constructed of thin glass,
and the base of the vessel made to slope so that the test-tube when
placed in the vessel has a tendency to be near the front. The
vessel is filled with a mixture of the same refractive index as the
nutrient gelatine. The latter has a refractive index rather higher
than water, which is about 1:333; alcohol has a refractive index of
1-374, The vessel is filled with water, and alcohol is then added
until the proper density is reached. The test-tube is placed in the
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 107
vessel, and held in position by means of a clip. The vessel can be
fixed on the inclined stage of the microscope, and the contents of the
tube conveniently examined with low-power objectives.
Plate-cultivations.—By this method, as already mentioned, a
mixture of bacteria, whether in fluids, excreta, or in cultivations on
solid media, can be so treated that the different species are isolated
one from the other, and perfectly pure cultivations of each of the
cultivable bacteria in the original mixture established in various
nutrient media. We are enabled also to examine under a low
power of the microscope the individual colonies of bacteria, and to
distinguish by their characteristic appearances, micro-organisms
which, in their individual form, closely resemble one another, or
are even identical. The same process, with slight modification, is
also employed in the examination of air, soil, and water, which will
be referred to later,
The preparation
of plate-cultivations,
therefore, must be
described in every de-
tail: and to take an
example, we will sup-
pose that a series of
plates is to be pre-
pared from a test-
cube-cultivation.
Fic. 36.—LEvELLING APPARATUS.
Arrangement of
Levelling Apparatus.—In order to spread out the liquid jelly
evenly on the surface of a glass plate, and hasten its solidifica-
tion, it is necessary to place the glass plate upon a level and
cool surface. This is obtained in the following manner: Place a
large shallow glass dish upon a tripod stand, and fill it to the brim
with cold water; carefully cover the dish with a slab of plate-glass,
or a pane of window-glass, and level it by placing the spirit-level in
the centre and adjusting the screws of the tripod. Substitute for
the spirit-level a piece of filter-paper the size of the glass plates to
be employed, and cover it with a shallow bell-glass.
Sterilisation of Glass Plates.—The glass plates are sterilised in
an iron box placed in the hot-air steriliser, at 150° C., from one to
two hours. As these plates are used also for other purposes, a
quantity ready sterilised should always be kept in the box.
Preparation of Damp Chambers.—The damp chambers for the
reception of the inoculated plates are prepared thus: Thoroughly
108 BACTERIOLOGY.
cleanse and wash out with 1 in 20 earbolic acid a shallow glass dish
and bell. Cut a piece of filter-paper to line the bottom of the glass
dish, and moisten it with the same solution.
Method of Inoculating the Test-tubes.—In
i a glass beaker or an ordinary glass tumbler,
with a pad of cotton-wool at the bottom,
place the tube containing the cultivation,
the three tubes to be inoculated, three glass
rods which have been sterilised by heating
in the flame of a Bunsen burner, and a
thermometer. Provide a strip of paper, a
Rie a7 bee Mee large label, a pencil, a pair of forceps, and
Gracy Iwas, inoculating needles. All is now ready at
hand to commence the inoculation of the tubes.
Liquefy the gelatine in the three tubes by placing them in a
beaker containing water at 30° C., or by gently warming them in
the flame of the Bunsen burner. Keep the tubes, both before and
after the inoculation, in the warm water, to maintain the gelatine
in a state of liquefaction. Hold the tube containing the cultivation
Fic. 38.—Metuop or Inocunatinc TEstT-TUBES IN THE PREPARATION
or PLATE-CULTIVATIONS.
and a tube of the liquefied gelatine as nearly horizontal as possible
between the thumb and index finger of the left hand. With the index
finger and thumb of the right hand loosen the plugs of the tubes:
Take the looped platinum needle in the right hand and hold it like
a pen. Remove the plug from the culture-tube by using the fourth
and fifth fingers of the right hand as forceps, and place it between
DESCRIPTION OF PLATE III.
Plate-cultivation.
This represents the appearance of a plate-cultivation of the comma-bacillus
of Cholera nostras, when it is examined overa slab of blackened plate-glass.
The drawing was made from a typical result of thinning out the colonies by
the process of plate-cultivation. At this stage they were completely isolated
one from the other; but later they became confluent, and produced complete
liquefaction of the gelatine.
Plate TIL
H. Crovkshankfacit
Vincent Brocks,Day & Son, Lith
PLATE-CULTIVATION.
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 109
the fourth and fifth fingers of the left. Remove the plug of the
other tube in the same way, placing it between the third and fourth
fingers of the left hand. With the needle take up a droplet of the
cultivation and stir it round in the liquefied jelly. Replace the plugs,
and set aside the cultivation. Hold the freshly inoculated tube be-
tween the index finger and thumb of either hand, almost horizontally,
then raise it to the vertical, so that the liquid gelatine gently flows
back. By repeating this motion and rolling the tube between the
fingers and thumbs the micro-organisms which have been introduced
are distributed throughout the gelatine. Any violent shaking, and
consequent formation of bubbles, must be carefully avoided. From
the inoculated tube, in the same manner inoculate a fresh tube of
liquefied gelatine, introducing into it three droplets with a sterilised
needle. After tilting and rolling this tube, as in the previous case,
the same process is repeated with a third tube, which is inoculated
from the second tube. This last tube must be inoculated in different
ways, according to experience, for different micro-organisms. Some-
times a sufficient separation of the micro-organisms is attained by
inoculating the last tube with a straight, instead of a looped, needle,
dipping it from the one into the other from three to five times.
The next process consists in pouring out the gelatine on glass
plates and allowing it to solidify.
Preparation of the Gelatine-Plates.—The directions to be observed
in pouring out the gelatine are as follows :—
Place the box containing sterilised plates horizontally, and so
that the cover projects beyond the edge of the table; remove the
cover, and withdraw a plate with sterilised forceps ; hold it between
the finger and thumb by opposite margins, rapidly transfer it to
the filter-paper under the bell-glass, and quickly replace the cover
of the box. On removing the plug from the tube which was first
inoculated, an assistant raises the bell-glass, and the contents of the
tube are poured on to the plate; with a glass rod the gelatine must
be then rapidly spread out in an even layer within about half an
inch of the margin of the plate. The assistant replaces the bell-
glass, and the gelatine is left to set. Meanwhile a glass bench or
metallic shelf is placed in the damp chamber, ready for the reception
of the plate-cultivation, and when the gelatine is quite solid the
plate is quickly transferred from under the bell-glaxs to the damp
chamber; precisely the same process is repeated with the other
tubes, and the damp chamber, labelled with the details of the
experiment, is set aside for the colonies to develop. Not only plate-
cultures should be carefully labelled with date and description, but
110 BACTERIOLOGY.
the same remark applies equally to all preparations—tube-cultureg,
potato-cultures, drop-cultures, etc.
Corresponding with the fractional cultivation of the micro-
organisms obtained in this manner, the colonies will be found to
develop in the course of a day or two, the time varying with the
temperature of the room. The lower plate will contain a countless
number of colonies which, if the micro-organism liquefies gelatine,
speedily commingle, and produce, in a very short time, a complete
liquefaction of the whole of the gelatine. On the middle plate the
colonies will also be very numerous, but retain their isolated position
for a longer time; while on the uppermost plate the colonies are
completely isolated from one another, with an appreciable surface of
gelatine intervening.
Examination of Plate-cultivations:—The macroscopical appear-
ances of the colonies are best studied by placing the plate on a
Fic. 39.—Dame CHAMBER CONTAINING PLATE-CULTIVATIONS.
slab of blackened glass, or on a porcelain slab if the colonies are
coloured.
To examine the microscopical appearances, a selected plate is
placed upon the stage of the microscope. The smallest diaphragm
is employed, and the appearances studied principally with a low
power. These appearances should he carefully noted, and a
sketch or photograph of the colony made. The morphological
characteristics of the micro-organisms of which the colony is formed
can be examined in the following way: A small looped or
hooked platinum needle is held like a pen, and the hand steadied
by resting the little finger on the stage of the microscope. The
extremity of the needle is steadily directed to the space between
the lens and the gelatine without touching the latter, until, on
looking through the microscope, it can be seen in the field, above
or by the side of the colony under examination. The needle
is then dipped into the colony, steadily raised, and withdrawn.
Without removing the eye from the microscope this manipulation
can be seen to be successful by the colony being disorganised or
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 111
completely removed from the gelatine. It is, however, not easy to
be successful at first, but with practice this can be accomplished
with rapidity and precision. A preparation is then made by rubbing
S88 SSa==
| ff
| l
Fic. 40.—Pasteur’s Larce INcuBATOR.
the extremity of the needle in a droplet of water on a slide, covering
with a cover-glass, and examining in the fresh state, or by spreading
out the droplet on a cover-glass, dvying, passing three times through
the flame, and staining with a drop of fuchsine or gentian violet.
Tnoculations should be made in test-tubes of nutrient gelatine
1 BACTERIOLOGY.
and agar-agar, from the micro-organisms transferred to the cover-
glass before it is dried and stained, from any remnants of the colony
which was examined, or from other colonies bearing exactly similar
appearances. In this way pure cultivations are established, and
the macroscopical appearances of the growth in test-tubes can be
obtained. The plates should be replaced in the damp chamber
as soon as possible; drying of the gelatine, or contamination with
micro-organisms gravitating from the air during their exposure,
may spoil them for subsequent examination.
A much simpler method of plate-cultivation
ix to dispense with the levelling apparatus, and
pour the liquefied jelly into shallow, flat dishes.
Fic. 41.—PeErtri’s
Disu.
They take up much less room, and in many
ways are more convenient (Fig. 41).
Nutrient agar-agar can also be employed for the preparation
of plate-cultivations, but it is much more difficult to obtain satis-
factory results. The: test-tubes of nutrient agar-agar must be
placed in a beaker with water and heated until the agar-agar
is completely liquefied. The gas is then turned down, and the
temperature of the water allowed to fall until the thermometer
stands just above 50° C. The water must be maintained at this
temperature, and the test-tubes must be in turn rapidly inoculated
and poured out upon the glass plates, or better still, into glass
dishes, as already described.
A very much simpler plan is to liquefy the agar, pour it into
Fic. 42.—Giass BENcHES AND SLIDEs.
a shallow dish, and allow it to solidify. The culture material is
thinned out in sterilised broth, and a few drops are spread out over
the surface of the agar. The dishes are then placed in the incubator
at 37° C.
Glass plates may also be employed in a much simpler way.
The nutrient jelly is liquefied, poured out, and allowed to set. A
needle charge with the material to he inoculated is then drawn
in lines over the surface of the jelly. This method is of use for
inoculating different organisms side by side, and watching the effect
of one upon the other, or a micro-organism in this way may be
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 113
sown upon the gelatine which has been already altered by the
growth of another micro-organism; the change produced in the
gelatine, as in the case of the Bacillus pyocyaneus, extending far
beyond the limits of the growth itself.
Nutrient jelly may also be spread out on sterilised glass slides,
which after inoculation are placed in damp chambers for the growths
to develop.
Esmarch’s Roll-cultures.—Esmarch introduced a modification
of the method of plate-cultivation which may sometimes be used with
advantage. The ordinary test-tubes may be employed, or tubes
considerably larger in size.
After the liquid jelly has been inoculated in the tube, instead
of pouring it out on toa glass plate or into a dish, the cotton-wool
plug is replaced, and an india-rubber cap fitted over the mouth of
the tube.
The tube is then placed horizontally on a block of ice or
in a vessel containing iced water. The neck of the tube is steadied
with the left hand, and the tube turned round and round with the
right hand. In a very short time the gelatine sets, and the tube
is lined inside with a thin coating. ‘There is far less danger of
contamination, and the cultures are in a much more convenient
form when circumstances render it necessary to move them.
(c) PREPARATION AND EmpitoymMEent oF Soipirrep Bioop Serum.
Solid Blood Serum.—The tubercle-bacillus, the bacilli of
glanders and of diphtheria, and many other micro-organisms, thrive
well when cultivated on solid blood serum. This medium has the
additional advantage of remaining solid at all temperatures. The
technique required for its preparation and sterilisation is as follows :
Several cylindrical vessels, about 20 cm. high, are thoroughly washed
with carbolic acid (1 in 20), and then with alcohol, and finally
rinsed out with ether. The ether is allowed to evaporate, and the
vessels are then ready for use. The skin of the animal selected—
calf, sheep, or horse—is washed with carbolic at the seat of operation,
and the bleeding performed with a sterilised knife or a trocar and
cannula. The first jet of blood from the vein is rejected, and that
which follows is allowed to flow into the vessels until they are almost
full. The ground-glass stoppers, greased with vaseline, are replaced,
and the vessels set aside in ice, as quickly as possible, for from
twenty-four to thirty hours. By that time the separation of the
clot is completed, and the clear serum can then be transferred to
8
114 BACTERIOLOGY.
plugged sterilised test-tubes. These should be filled, with a sterilised
pipette, to about one-third of their capacity.
Formerly the tubes were sterilised by Tyndall’s process of dis-
continuous sterilisation. The tubes were placed in Koch’s serum
steriliser, with the temperature maintained for an hour or more at
56° C., and this was repeated for six successive days, the temperature
on the last day being gradually raised to 60°C. ‘This completed the
sterilisation, and to solidify the serum the tubes were arranged in
the inspissator at the angle required, and the temperature was kept
between 65° C. and 68° C.
Directly solidification took
place the tubes were removed.
The new process is much
less tedious, and consists in
taking every possible pre-
caution to obtain the blood
without contamination by bac-
teria in the air or in the
vessels employed. There is
then no need to sterilise the
serum, and it can be coagu-
lated immediately. The tubes
are tested by placing them
in an incubator at 37° C. for
a week, and if any show
signs of contamination they
are discarded, and the rest
Fic. 43.—Kocn’s Serum STERILISER. can be used or kept in stock.
The serum should then
present the character of being hard, solid, of a pale straw colour,
and transparent. A little liquid collects at the lowest point, and
the serum is sometimes milky in appearance at its thickest part.
Liffler’s Blood Serum is prepared by mixing two-thirds of fresh
serum with one-third of broth, prepared in the usual way but with
the addition of 1 per cent. grape-sugar. The mixture is decanted
into test-tubes, avoiding the formation of air-bubbles, and it is then
coagulated in the usual way. The serum may be employed not only
in test-tubes, but also in small flasks, glass capsules, or other vessels,
all of which must be cleansed and sterilised.
Hydrocele fluid and other serous effusions may be prepared in
the same manner. Gelatine may be added to the serum in the pro-
portion of 5 per cent.
NUTRIENT MEDIA AND METHODS OF CULTIVATION, 115
Inoculation of the Tubes.—A small portion of a culture or of the
aterial to be inoculated is taken up with a sterilised platinum
needle, and traced over the sloping surface of the serum; or a
fragment of tissue, such as diphtheritic membrane or tubercle, may
be introduced into the tube and rubbed gently over the serum so as
not to break the surface.
EA
Fic. 44.—Hueppe’s Serum INSPIssaTor.
(D) PREPARATION AND EmpPLoyMEnt oF Srerinisep Poraro.
Potato-cultivations._Sterilised potatoes form an excellent
medium for the cultivation of many micro-organisms, more especially
the chromogenic species. Potato-cultivations also give in some cases
very characteristic appearances, which are of value in distinguishing
bacteria which possess morphological resemblances.
Preparation of Sterilised Potatoes.—Potatoes, preferably smooth-
skinned, which are free from ‘‘eyes” and rotten spots, should be
selected. If they cannot be obtained without eyes and spots, these
must be carefully picked out with the point of a knife. The potatoes
are well scrubbed with a stiff brush, and allowed to soak in | in 20
carbolic for a few minutes. They are then transferred to the potato-
116 BACTERIOLOGY.
receiver, and steamed in the steam steriliser for twenty minutes to
half an hour, the time varying according to the size of the potatoes,
When cooked, the potato-receiver is withdrawn and left to cool, the
potatoes being retained in it until required for use.
Damp chambers are prepared ready for the potatoes, the vessels
being cleansed and washed with carbolic as described for plate-
cultivations. Small glass dishes of the same pattern as the large
ones may be employed for single halves of potatoes. Potato-knives
and sealpels, which have been sterilised in an iron box by heating
them in the hot-air steriliser at 151° C. for one hour, should
be ready to hand. Knives sterilised by heating them in the flame
of a Bunsen burner should afterwards be placed upon a sterilised
glass plate and covered with a bell-glass. It must not be forgotten,
Fic. 45.—Box ror STERILISING INSTRUMENTS.
however, that heating the blades in the flame destroys the temper
of the steel, and therefore knives and other instruments should
preferably be sterilised in the hot-air steriliser, enclosed in an iron
box, or simply enveloped in cotton-wool.
Inoculation of Potatoes.—The coat-sleeves should be turned back,
and the hands, after thorough washing with good lathering soap,
be dipped in 1 in 40 carbolic. An assistant opens the potato-
receiver, and a potato is selected and held between the thumb and
index finger of the left hand. With the knife held in the right
hand, the potato is almost completely divided in the direction
which will give the largest surface. The assistant raises the cover
of the damp chamber, and the potato is introduced, and while the
knife is withdrawn, allowed to fall apart. The cover is quickly
replaced, and another potato treated in the same way, is placed
in the same damp chamber. The four halves are then quite ready
for inoculation. As an extra precaution, the left hand is again
dipped in carbolic, and one half of a potato is taken up between the
tips of the thumb and index finger, care being taken to avoid
touching the cut surface. Holding it with its cut surface vertical,
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 117
a small portion of the substance to be inoculated is placed on the
centre with a sterilised platinum needle. With a sterilised scalpel
the inoculated substance is rapidly spread over the surface of the
potato with the flat of the blade, to within a quarter of an inch
of the margin, and the potato is then as quickly as possible replaced
in the damp chamber. With another sterilised scalpel a small
portion of the potato from the inoculated surface of the first half
is in the same way spread over the surface of the second half, thus
thinning out the bacteria as in plate-cultivations. LHxactly the
same is repeated with a third potato, and even a fourth, so that
a still further thinning out or fractional cultivation of the micro-
organisms may be obtained. In some cases it is necessary to place
the cultures in an incubator (Fig. 40); others grow very well at the
Fic. 46.—Dame CHAMBER FOR POTATO-CULTIVATIONS.
temperature of the room. As in plate-cultivations, the potato may
also be inoculated by simply streaking it in lines with a needle
charged with the material to be cultivated.
Potato in Test-tubes.—Large surfaces of potato are employed when
we wish to obtain cultures of micro-organisms in considerable quanti-
ties, as in the examination of the products of chromogenic bacteria ;
but under ordinary circumstances potato 1s employed in test-tubes.
The central portions of raw potatoes are cut out in cylindrical pieces
with a cork-borer. ‘These are divided obliquely in their whole
length, and each half is placed in a test-tube. The test-tubes are
plugged with cotton-wool, and then steam in the steam-steriliser
for twenty minutes. The sloping surface is inoculated in the same
way as obliquely solidified jelly, and the advantages are great. The
cultures are obtained in a more convenient form, and there is less
danger of contamination.
Potato-paste may be employed when it is desirable to obtain an
extensive growth of certain bacteria. The potatoes are boiled for
an hour, and the floury centre squeezed out of the skins. This
is then mashed up with sufficient sterilised water to produce a thick
118 BACTERIOLOGY.
paste, and is heated in the steam steriliser for half an hour for three
successive days.
(e) PREPARATION AND EmpLoyMenT or Breap-Pasre, VEGETABLES,
Fruit, WuHite oF Eaa.
Some micro-organisms, more especially mould fungi, grow very
well on bread-paste. This is prepared by removing the crust from
slices of bread and drying them in the oven. They are then
broken up, and reduced to a fine powder with a pestle and mortar.
Small, carefully cleansed, conical, or globe-shaped flasks are plugged
with cotton-wool and sterilised in the oven. When cool, a small
quantity of the powder is placed in them, and sterilised water added
in the proportion of one part to every four of the powder. The
paste is sterilised by steaming in the steriliser at 100° C. for half an
hour for three successive days. The flasks can be reversed, and may
be inoculated with a platinum needle.
Boiled carrots and other vegetables, and various kinds of stewed
fruit, are also occasionally employed for the cultivation of bacteria.
The sterilisation of these media must be carried out on the principles
already explained.
White ‘of egg may be solidified in shallow glass dishes, in the
steam steriliser. After inoculation the dishes should be placed
in a damp chamber.
Liquip Mepis.
(F) Preparation oF STERILisED Broru, Liguip Broop Serum,
Urine, Mitx, VecEraste Inrusions, AND ARTIFICIAL NourRIsH-
inc Liquips.
Nutrient liquids are still largely employed. For inoculation ex-
periments when the presence of gelatine is undesirable, for studying
the physiology and chemistry of bacteria, and when for any object
a rapid growth of micro-organisms is necessary, the employment
of liquid media is not only advisable, but absolutely necessary.
Liquid media comprise two distinct groups—natwral and artificial.
Natural media include meat broth, blood, urine, milk, and vegetable
infusions ; artificial media are solutions composed from a chemical
formula representing essential food constituents.
Broth may be made from beef, pork, chicken, or fish in the
manner which has been described for the preparation of nutrient
gelatine, simply with omission of the gelatine. After the process
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 119
of neutralisation with carbonate of soda solution, the flask of broth
is placed in the steam steriliser for half an hour at 100°C. A
clear liquid results on filtration which is transferred to plugged
sterilised flasks or test-tubes, and sterilisation effected by exposing
them in the steam steriliser for half an hour at 100° C. for two
eM manal
«TT um
Fic. 47.—APPARATUS FOR STERILISATION BY STEAM UNDER PRESSURE.
or three successive days, or by using the apparatus for sterilising
by steam under pressure. For some bacteria a more suitable
cultivating medium is obtained by the addition of glycerine or
grape-sugar.
Liquid Blood Serum.—The preparation of blood serum has
already been described. It may be required for cultivations before
the final treatment -by; which it is solidified, for example, in the
method of drop-cultivation, and may be used with the addition of
glycerine or grape-sugar. Hydrocele fluid, peritonitic and pleuritic
120 BACTERIOLOGY.
effusions, can also be employed after sterilisation in the steam
steriliser. The fluid should be withdrawn with a sterilised trocar
and cannula, and received into plugged sterilised flasks.
Urine.—In order to obtain urine free from micro-organisms the
following precautions must be observed: The orifice of the urethra
must be thoroughly cleansed with weak carbolic. The first jet of
urine should be rejected, and the rest received into sterilised vessels,
which must be quickly closed with sterilised plugs. If these pre-
cautions be not attended to, the urine must be rendered sterile by
the means described for the sterilisation of broth.
Milk.—If milk has been drawn into sterile flasks, after thoroughly
cleansing and disinfecting the teats and hands, it may be kept
without change. If procured without these precautions, it must
be steamed in the steriliser for half an hour for five successive
days.
Vegetable and other Infusions.—Infusions of hay, cucumber,
and turnip are used for special purposes, and more rarely decoctions
of plums, raisins, malt, and horse-dung. They are mostly prepared
by boiling with distilled water, after maceration for several hours.
The filtrate is received into sterile flasks and sterilised in the usual
way in the steam steriliser.
Artificial Fluids.—Pasteur’s solution is prepared by mixing the
ingredients in the following proportions :—
Distilled water . 160
Pure cane-sugar 10
Ammonium tartrate : 1
Ash of yeast : : 075
Mayer’s modification of the nourishing fluid employed by Cohn is
as follows :— ,
Distilled water. 20
Ammonium tartrate “D
Phosphate of potassium ‘ : ‘1
Sulphate of magnesium ‘1
Tribasic calcium phosphate 01
Drop-cultures.—This method of cultivation is a particularly
instructive one. It enables us to study many of the changes which
take place during the life history of micro-organisms. This is
illustrated, for example, in a drop-culture of the anthrax bacillus,
in which we can watch the gradual growth of a single bacillus into
NUTRIENT MEDIA AND METHODS OF CULTIVATION, 121
a long filament, and the subsequent development of bright oval
spores. It is necessary carefully to observe the minutest details
in order to maintain the cultivation pure. An excavated slide is
thoroughly cleaned, and then sterilised by being held with the
cupped side downwards in the flame of the Bunsen burner. A ring
of vaseline is painted round the excavation, and the slide is then
placed under a glass bell. Meanwhile a carefully cleansed cover-
glass is also sterilised by passing it through the flame, and should
be deposited on a sterilised glass plate. With a sterilised looped
needle, a drop of sterile broth is transferred to the cover-glass,
and this is inoculated by touching it with another sterilised needle
charged with the material to be examined, without disturbing
the form of the drop. It is quite sufticient just to touch the drop
h a,
Fic. 48.—Drop CUuLtivaTion.
(a) Drop of broth; (b) layer of vaseline.
instead of transferring a visible quantity of blood, juice, or growth,
as the case may be. The slide is then inverted and placed over the
cover-glass, so that the drop will come exactly in the centre of the
excavation, and is gently pressed down. On turning the slide over
again the cover-glass adheres, and an additional layer of vaseline
is painted round the edges of the cover-glass itself. The slide must
be labelled, and if necessary, placed in the incubator, and the results
watched from time to time. Instead of broth, liquid blood serum
may be employed in this form of cultivation. If it is required
to preserve the drop-cultivation as a microscopic preparation, the
cover-glass is gently lifted off and allowed to dry. Any vaseline
adhering to the cover-glass should be wiped off, and the cover-glass
can then be passed through the flame and stained in the usual manner.
Moist Cells.—Unless drop-cultures are very carefully prepared
122 BACTERIOLOGY.
they are liable to dry up, if kept for examination for several days.
Many therefore prefer employing a moist cell, of which there are
several different forms in use.
The drop-culture slide may be converted into a moist cell
by having a deep groove cut round the circumference of the con-
cavity. This groove is filled with sterilised water by means of a
pipette. A ring of vaseline is painted with the camel’s-hair brush
outside the groove, and the cover-glass, with the drop-cultivation,
is inverted and placed over the concavity. This form is very useful,
as the slide can be easily cleansed and effectually sterilised by
holding it in the flame of the Bunsen burner.
A very simple form of moist cell recommended by Schafer
Fic. 49.—SmureteE Meruop or Formuine a Moist CELL.
may be used in some cases, but possesses the disadvantage of not
admitting of sterilisation by heat. A small piece of putty or
modelling wax is rolled into a cord about two inches long and 3 inch
thick. By uniting the ends a ring is formed, which is placed on the
middle of a clean glass slide. A drop of water is placed in the
centre of the ring, and the cell roofed in by applying a cover-glass.
A cell somewhat similar in form, which has the advantage of
permitting of thorough cleansing, may be constructed by cementing
a glass ring with flat surfaces to an ordinary slide. Vaseline is
applied with a camel’s-hair brush to the upper surface of the ring,
and one or two drops of water placed with a pipette at the bottom
of the cell. The cover-glass, with the preparation, is then inverted
over the cell and gently pressed down upon the glass ring. The
vaseline renders the cell air-tight, and, to a certain extent, fixes
the cover-glass to the ring.
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 123
Warm Stages —To apply warmth while a preparation is under
continuous observation, we must either place the microscope bodily
fa SE
Fig. 50.—WarM STAGE.
within a special incubator, with the eye-piece protruding through an
opening, or we must employ some means of applying heat directly
to the preparation.
A simple warm stage may be made of an oblong copper plate,
Fic, 51.—WarM STAGE SHOWN IN OPERATION.
two inches long by one inch wide, from one side of which a rod of
the same material projects. The plate has a round aperture in the
124 BACTERIOLOGY.
middle, half an inch in diameter, and is fastened to an ordinary
slide with sealing-wax. The drop to be examined is placed on a
large-sized cover-glass and covered with a smaller one. Olive oil
or vaseline is painted round the edge of the smaller cover-glass to
prevent evaporation, and the preparation is placed over the aperture
in the plate.
The slide bearing the copper plate is clamped to the stage of
Fic. 52.—IsRaAEL’s WARMING APPARATUS IN OPERATION.
the microscope. The flame of a spirit-lamp is applied to the
extremity of the rod, and the heat is conducted to the plate and
thence transmitted to the specimen. In order that the temperature
of the copper plate may be approximately that of the body, the lamp
is so adjusted that a fragment of cacao butter and wax, placed close
to the preparation, is melted.
Israel’s Warming Apparatus.—It is obvious that in employing
very high powers a difficulty will be presented by the warm stages
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 125
commonly used for accurate observations, such as Schifer’s or
Stricker’s, owing to their interference with the illumination. To
Fic, 53.—SxecTion or Israkt’s WARMING APPARATUS AND Drop-cULTuRE
SLIDE.
overcome this an apparatus has been constructed by which the
slide is warmed from above (Figs. 52, 53).
The drop-culture slides are provided with a shallow groove, ‘1 mm.
deep and 1 mm. broad, cut round the concavity. Into this the
cover-glass fits, so that its upper surface is level with that of
Fic. 54.—IsRAEL’s WARMING APPARATUS.
the slide. The heating apparatus consists of a flat disk-shaped box
with a central conical aperture.
The entrance and exit pipes are fixed on at a right angle to the
126 BACTERIOLOGY.
side (Fig. 54). The former, z, is of metal, and,the latter, a, of glass
fitted with a thermometer, the bulb of which, &, is contained within
the box. A partition, s, keeps up a current between the openings
Fic. 55.—GAs CHAMBER IN USE WITH APPARATUS FOR GENERATING
Carponic ACID.
of the pipes, which are supported on a stand and connected by
tubing with the hot-water supply.
A mixture of paraffine and vaseline is recommended for indicating
the temperature of the chamber, and experience has shown that if
a temperature of 37° C. is required the temperature of the water
in the box must range between 42° and 47° C.
Fic. 56.—GAs CHAMBER.
Gas Chambers.—To investigate the action of gases or vapours
upon micro-organisms, a modification of the moist cell may be
employed.
A piece of glass tubing is first fixed to the slide by means of
NUTRIENT MEDIA AND METHODS OF CULTIVATION, 127
sealing-wax, and the ring of putty is so placed as to include one end
of it, leaving a small interval at the side, or a little notch is made
in the putty opposite, so as to afford an exit for the gas or vapour.
Fic. 57.—Moist CELL ADAPTED FOR TRANSMISSION OF ELECTRICITY.
Application of Hlectricity.—To study the effect of electricity
we may prepare a drop-culture in the moist cell. The cover-
glass to be used is provided with two strips of tinfoil, which are
Fic. 58.—APPARATUS ARRANGED FOR TRANSMITTING ELECTRICITY.
isolated from the brass of the microscope, and so arranged that a
current of electricity may be passed through them.
A much simpler plan, which may also be employed, is to take
an ordinary glass slide and coat the surface with gold-size. The
128 BACTERIOLOGY,
slide is then pressed firmly down on gold-leaf or tinfoil and allowed
to dry. When dry, the metal is scraped away, leaving two triangles
with a small interval between them.
Fic. 59,—Simr WITH GOLD-LEAF ELECTRODES.
The liquid containing the micro-organisms is placed between the
electrodes, covered with a cover-glass, and then subjected to the
electric current.
(@) Mernops or Empioyine AND Srorine Liquip Mepzia.
Cultivations in liquid media can be carried on in test-tubes, but
it is more satisfactory to employ special forms of flasks, bulbs, and
U tubes, such as those employed by Pasteur and his school, and by
Lister, Sternberg, and Aitken.
Lister's Flasks.—These flasks were especially introduced by Lister
as a means of storing liquid nutrient media.
They are so constructed that after removal
of a portion of the contents, on restoring
the vessel to the vertical position, a drop of
liquid always remains in the extremity of
the nozzle, which prevents regurgitation of
unfiltered air.
Sternberg’s Bulbs.—The method of intro-
ducing liquid into the bulb employed by
Fic. 60.—ListeR’s Sternberg, and of sterilising and inoculating
ess it, is as follows: The bulb is heated slightly
over the flame, and the extremity of the neck, after the sealed point
has been broken off, is plunged beneath the surface of the liquid.
As the air cools the liquid is drawn into the bulb,
usually filling it to about one-third of its capacity. (=
The neck of the flask is again sealed up, and the
liquid which has been introduced is sterilised by
repeatedly boiling the flasks in the water-bath,
They should then be placed in the incubator for two or three days ;
Fic. 61.—STERN-
BERG’S Bus.
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 129
and if the contents remain transparent and free from film, they
may be set aside as stock-bulbs, to be used when required.
To inoculate the liquid in the bulb the end of the neck is heated
to sterilise the exterior, the bulb is gently warmed, and the extremity
of the neck nipped off with a pair of sterilised forceps. The open
extremity is plunged into the liquid containing the micro-organisms,
and a minute quantity enters the tube and mingles with the fluid in
the bulb without fear of contamination by atmospheric germs, The
extremity of the neck is once more sealed up in the flame of a
Bunsen burner.
Aitken’s Tubes.—These tubes are plugged and sterilised, and the
nutrient medium introduced as into ordinary test-tubes. Instead
of withdrawing the cotton-wool plug, they are
inoculated through a lateral arm. The sealed
extremity of the arm is nipped off with
sterilised forceps, and the inoculating needle is
carefully introduced through the opening thus
made. It is directed along the arm until it
touches the opposite side of the test-tube, where
Fic. 62.—AITKEN’s —_ it, deposits the material with which it was charged.
Tee The needle is withdrawn, and the end of the
lateral arm again sealed up in the flame; the test-tube is then
tilted until the liquid touches the deposited material; on restoring
the tube to the vertical, the material is washed down with the
nutrient liquid.
Miquel’s Bulbs.—The tube a 7
boule of Miquel is also a very
useful form. It consists of a
bulb of 50 cc. capacity, blown
in the middle of a glass tube.
The part of the tube above the
bulb is contracted in two places,
and can either be left quite
straight or made to curve
slightly. Between the contrac-
tions the tube is plugged with
asbestos. The portion of the
tube below the bulb is S shaped, Fic. 63.—Mr1quet’s Bute.
and drawn out at its extremity
into a fine point. The bulb is charged with nutrient liquid and
inoculated by aspiration, and the point of the § tube sealed in the
flame of a Bunsen burner.
9
130 BACTERIOLOGY.
Pasteur’s Apparatus.—Special forms of tubes, bulbs, and pipettes
are employed by the school of Pasteur. The tubes are provided
with lateral or with
curved arms drawn
out to a fine point,
and with slender
necks plugged with
cotton-wool. A
double form (Fig. 65)
shaped like a tuning-
fork, each limb with
a bent arm, is con-
Fic. 64,—PAsTEvR’s Fiask. venient for storing
. sterilised broth. The
sealed end of an arm is nipped off with sterilised forceps, the sterile
broth aspirated into each limb, and the arm again sealed in the
flame; a series of such tubes can be
arranged upon a rack on the working
table.
Bulbs with a vertical neck drawn out
to a fine point, others with a neck bent
at an obtuse angle, plugged with cotton-
wool, and a lateral curved arm drawn
out to a fine point, are also employed.
For a description of these various vessels
and their special advantages, the works
of Pasteur and Duclaux must be con-
Fic. 65.—Pastevr’s DouB.e
sulted. =
TUBE.
(4) Cunrivation or ANAEROBIC BacrTERta.
To cultivate anaerobic organisms the same media are employed
as for aerobic organisms, but the methods must be modified, or special
apparatus used, so that the oxygen in the air may be excluded.
In the preparation of plate-cultivations, before the film of gelatine
has completely hardened it is covered with a sheet of mica, and the
edges are sealed with melted paraffine. By this process the air is not
completely excluded, so that only those organisms which are not
strictly anaerobic can be grown by this method. Liborius recom-
mends boiling a considerable volume of gelatine in a tube, cooling it,
and after thoroughly distributing the organisms in the still liquid jelly,
rapidly solidifying it by placing the tube in iced water. By this
NUTRIENT MEDIA-AND METHODS OF CULTIVATION. 131
process very little air re-enters the jelly, and colonies of even strictly
anaerobic bacteria will develop in the lower part of the tube. The
drawback is the difficulty encountered in examining the colonies,
and in preparing sub-cultures. For this purpose the tube
must be broken, or carefully warmed until the jelly can be
shaken out.
Esmarch first prepares a roll culture, and when the gelatine film
has set, the tube is completely filled with liquefied gelatine which has
been cooled down almost to the temperature at which it solidifies. The
same difficulty arises as
in the previous method, in
the examination of the
colonies.
Buchner places -the
culture tube inside a much
larger tube containing a
small quantity of pyro-
gallic acid and closed with
a gutta-percha cap. The
pyrogallic acid absorbs the
oxygen, but the method
is not altogether success-
ful.
The most satisfactory
plan is to exhaust the air
with an air pump, or to
substitute an atmosphere
of hydrogen which does
not affect the growth of
the bacteria.
Various forms of flasks
tae vale for cultivating Fic. 66.—FRANKEL’s ANAEROBIC TUBE-CULTURE™
bacteria have been devised, aa, Glass tube through which hydrogen is
which can be easily con- passed; 0, exit tube; e, india-rubber
nected with an exhausting stopper coated externally with paraffin
: . (FRANKLAND).
apparatus, and readily .
sealed by the flame of the blowpipe when the air has been removed.
If hydrogen is employed the most convenient plan is to use
a Kipp’s apparatus, from which the hydrogen is passed through
two bottles, one containing a solution of lead, to remove any
sulphuretted hydrogen, and the other pyrogallic acid, to intercept
any oxygen.
132 BACTERIOLOGY.
In the method recommended by Frankel a tube of gelatine is
liquefied, and inoculated. A gutta-percha stopper is substituted
for the cotton-wool plug (Fig. 66). It is perforated by two holes,
through which two tubes pass which are bent at a right angle.
One tube only just passes through the stopper, the other reaches
down to the bottom of the test-tube. The
horizontal part of each tube has a narrow
neck. The long tube has a plug of steril-
ised cotton-wool, and is connected with
a short piece of india-rubber tubing by
which it can be connected with Kipp’s
apparatus. The hydrogen drives the air
out of the liquefied jelly and out of the
test-tube, and after about half an hour
the horizontal tubes are sealed up, and
the test-tube is made into a roll culture.
Liborius employs a tube with a narrow
neck and a lateral arm (Fig. 67). The
tube is filled up to the height of the arm
with either nutrient agar or a mixture of
nutrient agar with 2 per cent. of grape-
sugar. The liquefied jelly is inoculated
in the usual way, and hydrogen passed
through the lateral arm. When the air
has been completely driven out, the tube
is sealed up.
To cultivate anaerobic organisms in
broth, such as the tetanus bacillus, a flask
is inoculated with the bacillus, and a
stream of hydrogen is passed through the
broth by means of a tube passing down to the bottom of the
flask. The air in the flask escapes by a lateral arm which is bent
down at a right angle, and immersed in a capsule of mercury.
When the air has been completely expelled the entrance tube is
hermetically sealed, and the mercury in the capsule prevents any
air from re-entering the flask by the lateral arm (Fig. 68).
Fic. 67.—ANAEROBIC CUL-
TURE-TUBE (LIBORIUS).
Meruop or Fixing Cuiturss.
The colonies in plate-cultivations and the growths of bacteria
in test-tubes may be stopped at any stage of their growth, and
permanently fixed by exposing the culture to the fumes of formic
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 133
aldehyde. The test-tubes, dishes, or capsules are placed in a cylin-
drical glass vessel containing a pledget of cotton-wool moistened with
Fic. 68.—APPARATUS FOR ANAEROBIC CULTURES.
(RoscoE anp Lunt.)
formic aldehyde. The vessel is fitted with a ground glass stopper
and set aside. The growth almost immediately ceases. Any
liquefied gelatine is hardened, so that the exact appearances of
cultures are obtained in a permanent form.
CHAPTER X.
EXPERIMENTS UPON THE LIVING ANIMAL.
To carry out the last of Koch’s postulates, and so complete the
chain of evidence in favour of the causal relation of micro-organisms
to disease, and to study the mode of action. of a pathogenic bacterium,
it is necessary to introduce into a living animal a pure cultivation
of the micro-organism or its chemical products. For this purpose
various animals are employed, such as mice, rabbits, guinea-pigs,
pigeons, and fowls.
Inhalation.—The animals may be made to inhale an atmosphere
impregnated with micro-organisms by means of a spray. In this -
way Friedlander succeeded in administering the bacteria of pneu-
monia to mice; and the production of tuberculosis by experimental
inhalation has thrown light upon the clinical records of cases
reported as instances of the infectiousness of phthisis.
Ingestion.— A sheep fed upon potatoes which have been the
medium for the cultivation of the anthrax bacillus dies in a few
days. Rabbits fed on cabbage sprinkled with a culture of the
bacillus of fowl cholera, rapidly succumb to the disease. Animals
fed upon the nodules of bovine tuberculosis or upon tubercular
flesh and milk will be readily infected.
Milk, or bread soaked in milk, is a very convenient medium,
and from a public health point of view, a most instructive way of
administering and testing the effect of pathogenic bacteria.
Vaccination and Subcutaneous Inoculation.—Vaccination may be
performed by making a few superficial scratches and inoculating the
wound with a sterilised platinum needle charged with the micro-
organisms. Another simple method is to take a sterilised scalpel,
infect the point with the material to be inoculated, and then make
a minute puncture or incision. In either case a situation should be
selected, such as the root of the ear, which cannot be licked by the
animal after the ope ation.
134
>)
EXPERIMENTS UPON THE LIVING ANIMAL. 135
Subcutaneous inoculation is very simple and effectual, and con-
sequently the method most frequently employed. The animal
selected—for example, a guinea-pig—is held by an assistant, who
covers it with a towel, leaving only the hind extremities exposed.
By so doing, and gently laying it upon its back, with its head low,
a guinea-pig passes apparently into a state of hypnotism, and the
Fic. 69.—Kocw’s SyRinceE.
trivial operation can be performed with little or no movement on the
part of the animal. From a spot on the inner side of the thigh the
hair is cut close with a small pair of scissors curved on the flat, and
the skin must be thoroughly purified with 1 in 20 carbolic acid. A
small fold of skin is then pinched up with a pair of sterilised forceps,
and with a pair of sharp sterilised scissors, or with a tenotomy knife,
a minute incision is made. A sterilised platinum loop is charged
with the material to be inoculated, and the loop is gently inserted
under the skin, forming a small pocket in the subcutaneous tissue.
The needle is then withdrawn, and the sides of the wound gently
pressed into apposition and painted over with collodion.
Fic. 70.—SyRINGE WITH ASBESTOS PLUG.
In inoculating a mouse the same process is adopted, with the
exception that the root of the tail is the usual site of the
operation.
In some cases it may be necessary to inoculate cultures diffused
in sterilised salt solution, or blood or lymph containing bacteria,
or a culture in broth, or a filtrate containing the toxic products,
136 ; BACTERIOLOGY.
and then a hypodermic syringe may be required. One of the ordinary
pattern may be used, but it is very much better to employ a syringe
which has been especially constructed to admit of thorough dis-
infection. Koch’s syringe is a convenient form, the liquid being
expressed by pressure on a rubber ball.
The author has generally preferred to improvise a substitute for
the hypodermic syringe which can be quickly made, and is destroyed
after use, so that there can be no possible risk of accidentally
infecting other animals. A short length of ordinary glass-tubing is
sterilised, and plugged at one end with sterilised cotton-wool ; about
three inches from the plug a bulb is blown about the size of a
marble, and two inches below this the glass is drawn out into a long
capillary tube. A sufficient quantity of the liquid to be injected rises
up into the tube by capillary attraction, or can be drawn up by
means of an india-rubber ball, until the bulb is full. The point of
.. the capillary tube is inserted through the opening in the skin, and
gently pushed into the subcutaneous tissue, and then withdrawn for
a short distance. By pressure on the bulb the contents of the tube
are injected. In dealing with chemical products there is no risk in
applying the lips and blowing out the contents of the tube, or indeed
of filling it by suction, for if too much force were applied the liquid
which might enter the mouth would be stopped by the cotton-wool
plug.
A number of these capillary tubes can be placed in a small case,
and when it is necessary to go to a distance to investigate an outbreak,
they will be found most convenient to bring back lymph or blood to
the laboratory for further study. .
Sternberg takes a piece of glass-tubing, blows a bulb at one end,
and draws out the other end into a thin tube. By heating the bulb
and then dipping the tube into the liquid to be inoculated the latter
rises in the tube as the bulb cools. After inserting the point of
the tube subcutaneously the bulb is again heated, and the liquid is
forced out into the tissues.
Intravenous Injection.—A cultivation of micro-organisms may be
mixed with sterilised water, and then injected with a syringe directly
into the circulation. This may be performed without much difficulty
by injecting, with a hypodermic syringe, the large vein at the base
of the ear in rabbits, or the jugular vein in large animals.
Special Operations.—In many cases it is absolutely necessary to
perform an operation of greater severity. After the administration
of an anesthetic, infective material may be inserted, or injected, into
the peritoneal cavity, or injected into the duodenum in the manner
EXPERIMENTS UPON THE LIVING ANIMAL. 137
employed in the case of Koch’s comma bacilli by Nicati and Rietsch.
In such cases antiseptic precautions must be rigidly followed, and
use made of iodoform and other antiseptic dressings. The disinfec-
tion of the skin of the animal, of the instruments employed, and of
the hands of the operator, are details essential to secure success.
To inoculate tubercular matter, sputum may be rubbed up with
distilled water, and some of the mixture injected into a tracheal
fistula; or the first steps of the operation of iridectomy may be
performed and tubercular material inserted into the anterior chamber
of the eye, but this method is only justifiable when it is absolutely
necessary for the results and changes to be observed from day
to day.
To inoculate rabbits or other animals with the virus of rabies,
the skull is trephined, and an emulsion prepared from the spinal cord
of a rabid animal is injected beneath the dura mater.
Before every inoculation the instruments must be sterilised in a
hot-air steriliser or by immersion in boiling water in a flat dish or
enamel tray heated by a spirit-lamp, and after each operation all
instruments should be placed in carbolic acid (1 in 20) or in boiling
water, wiped dry, and again sterilised in the hot-air steriliser, before
they are put away. If these precautions are not observed, instances
of accidental infection are sure to occur.
After the inoculation is completed a careful record must be made
of the date and details of the experiment. The form in which the
virus was used, the quantity employed, and the seat of inoculation,
must be taken into account. The animals must be kept under close
observation, the temperature taken, and any signs of illness, such as
ceasing to feed, difficulty in breathing, staring coat, and any local
signs, such as the development of a tumour or an enlargement of the
lymphatic glands, must be carefully noted.
It is perhaps hardly necessary to add that in this country no
experiments of any kind may be performed on living animals without
a license.
Meruop oF Dissection AND EXAMINATION.
All animals that die after an experimental inoculation should
be examined immediately after death. Every precaution must be
taken in conducting the dissection, to exclude extraneous micro-
organisms, and all instruments employed must have been sterilised
in the hot-air steriliser, or by immersion in boiling water. Ifa mouse,
for example, has died after inoculation with anthrax, it should be at
138 BACTERIOLOGY.
once pinned out by its feet on a slab of wood or in a gutta-percha
tray, and bathed with | in 40 carbolic. In the same way, before
examining a dead rabbit, a stream of carbolic should be directed
over it to lay the fur, which otherwise interferes with the dissection,
The hair should be cut away with sterilised scissors from the seat
of inoculation, which is the first part to be examined, and any
suppuration, hemorrhage, cedema, or other pathological change should
be carefully noted. From any pus or exudation that may be
present, material for inoculations should at once be taken, and
cover-glass-preparations made for microscopical examination.
To examine the internal organs and to make inoculations from
the blood of the heart or spleen, the skin is cut through from below
upwards in the median line of the abdominal and thoracic regions.
The abdominal cavity is then opened, and the walls pinned back.
on either side of the animal. Any abnormal appearances in the
peritoneum should be noted, and the state of the spleen should be
carefully examined by turning the intestines aside. After noting
its appearances, it should be removed with sterilised forceps and
scissors, and deposited upon a sterilised glass slide, and incised with
sterilised scissors. The cut surface is then touched with the point
of a sterilised inoculating needle, and cultures are made in test-tubes
of nutrient gelatine and agar-agar, and also on potato, and in broth
in the form of drop-cultivations. Precisely the same care must be
taken in examining lymphatic glands, tubercles, or pathological
nodules ; any chance putrefactive micro-organisms on the surface
should be destroyed by carbolic acid or the actual cautery; an
incision is then made, and a minute fragment snipped out of the
centre of the nodule, which can be inoculated in the living animal or
transferred to a cultivating medium.
The examination of the thorax is made by cutting through the
ribs on either side of the sternum with sterilised: scissors, and
turning the sternum up where it will be out of the way. The
pericardium is then opened, and the right auricle or ventricle pierced
with the point of a sterilised scalpel, and inoculations and -cover-
glass- preparations are made from the blood which escapes.
The lungs also require to be especially studied. ‘They should be
incised with a sterilised scalpel, and inoculations and cover-glass-
preparations made from the cut surface. It may be necessary to
embed a piece of lung or fragment of spleen, so that it shall be free
from air. This may be done by isolating a fragment with the
precautions just described, and depositing it upon the surface of a
test-tube of nutrient agar-agar. The contents of another tube,
EXPERIMENTS UPON THE LIVING ANIMAL. 139
which have been liquefied, and allowed to cool almost to the point
of gelatinisation, must then be poured over it. From a potato a
little cube must be cut, the tissue deposited in the trough thus
formed, and the cube replaced, or cultures may be prepared by any
of the methods which have been described for dealing with anaerobic
bacteria. Blood may also be taken directly from a vein by laying it
bare by dissection, making a small opening with sterilised scissors,
and inserting a looped platinum needle, the needle of a hypodermic
syringe, a capillary tube, or the extremity of the capillary neck of
a Sternberg’s bulb. If the cultivation, in spite of these precautions,
is contaminated, or if there was more than one organism present
in the blood or tissues under examination, it will be necessary to
separate the different kinds by plate-cultivation.
Having completed the dissection, the organs of such a small
animal as a mouse may be removed en masse, and transferred to
absolute alcohol for subsequent examination. In other cases it may
be only necessary to reserve portions of each organ. 1n experimenting
with a virulent micro-organism like anthrax, any remaining part of
the animal should be cremated, and the hands and all instruments
should be thoroughly disinfected.
Isolation of Micro-organisms during Life.—Micro-organisms in
the living subject may be isolated from the pus of abscesses, or
other discharges, and from the blood and tissues. Abscesses should
be opened, and other operations performed, when practicable, with
Listerian precautions, and a drop of the discharge taken up with
it looped needle or capillary pipette, as already explained.
To make a cultivation from the blood of a living person, the tip
of a finger must be well washed with soap and water and sponged
with 1 in 20 carbolic. Venous congestion is produced by applying
an elastic band or ligature to the finger, which is pricked with a
sterilised sewing needle. From the drop of blood which exudes the
necessary inoculations and examinations can be made. Another
way of extracting blood from the living patient is to apply a leech.
This method has been found of considerable value in experimenting
upon the blood of patients suffering from malaria, and may be
useful in other diseases, if the blood is required for further
examination, or in quantity.
CHAPTER XI.
EXAMINATION OF AIR, SOIL, AND WATER.
AIR.
TuE air, as is well known, contains in suspension, mineral, animal,
and vegetable substances. _ The mineral world is represented
by such substances as silica, silicate of aluminium, carbonate and
phospate of calcium, which may be raised from the soil by the
wind, and particles of carbon, etc., which gain | access from acci-
dental sources. Belonging to the animal kingdom we find the
débris of perished creatures, as well as, sometimes, living animals.
The vegetable world supplies micrococci, bacilli, and other forms
of the great family of bacteria, spores of other fungi, pollen seeds,
parts of flowers, and so forth. The air of hospitals and sick rooms
has been found to be especially rich in vegetable forms; fungi
and spores have been stated to ke present in particularly large
numbers in cholera wards; spores of tricophyton have been dis-
covered in the air of hospitals for diseases of the skin, and of achorion
in wards with cases of favus. The tubercle bacillus is said to have
been detected in the breath of patients suffering from phthisis. —
These points indicate that, in addition to the interest for the
micro-biologist, considerable importance, from a hygienic point of
view, must be attached to the systematic examination of the air.
A knowledge of the microbes which are found in the air of marshy
and other unhealthy districts, and in the air of towns, dwellings,
hospitals, workshops, factories, and mines, will be of practical value.
Miquel, who has particularly studied the bacteria in the air,
has found that their number varies considerably. The average
number per cubic metre of air for the autumn quarter at Mont-
souris is given as 142, winter quarter 49, spring quarter 85, and
summer quarter 105. In air collected 2,000 to 4,000 metres above
the sea-level,~not a single bacterium or fungus spore was found,
while in 10 cubic metres of air from the Rue de Rivoli (Paris) the
number was computed at 55,000.
140
EXAMINATION OF AIR, SOIL, AND WATER. 141
The simplest method for examining the organisms in air consists
in exposing plates of glass or microscopic slides coated with
glycerine, or with a mixture of glycerine and grape sugar, which is
stable, colourless, and transparent. Nutrient gelatine spread out on
glass plates may be exposed to the air for a certain time, and then
put aside in damp chambers for the colonies to develop. Sterilised
potatoes, prepared in the usual way, may be similarly exposed. In
both the last-mentioned methods separate colonies develop, which
may be isolated, and pure cultivations carried on in various other
nutrient media. Nutrient gelatine has also been employed in the
special methods of Koch and Hesse.
Koch's Apparatus.—This consists of a glass jar, about six inches
high, the neck of which is plugged with cotton-wool. In the
interior is a shallow glass capsule, which can be removed by means
of a brass lifter. The whole is sterilised by exposure to 150° C.
for an hour in the hot-air steriliser. The nutrient gelatine in a
stock-tube is liquefied, and the contents emptied into the glass
capsule. The jar‘is exposed to the air to be examined for a definite
time, the cotton-wool plug replaced, and the apparatus set aside for
the colonies to develop.
Hesse’s Apparatus.—The advantage of this apparatus is that it
enables the experimenter to examine a known volume of air. A
glass cylinder, 70 cm. long and 3°5 cm. in diameter, is closed at one
end by an india-rubber cap, perforated in the centre. Over this fits
another cap, which is not perforated. The opposite end of the
cylinder is closed with a caoutchouc stopper, perforated to admit
a glass tube plugged with cotton-wool. The tube can be connected
by means of india-rubber tubing with an aspirating apparatus,
which consists of a couple of litre-flasks, suspended by hooks from
the tripod-stand which supports the whole apparatus. The cylinder,
caps, and plug are washed with solution of carbolic acid, and
then with alcohol. After being thus cleansed, 50 cc. of nutrient
gelatine are introduced, and the whole sterilised by steaming for
half an hour for three successive days. After the final sterilisation,
the cylinder is rotated on its long axis, so that the nutrient medium
solidifies in the form of a coating over the whole of the interior.
When required for use, the cotton-wool plug is removed from the
small glass tube, and the latter connected with the upper flask by
means of the india-rubber tubing.
The apparatus is placed in the air which is to be examined, the
outer india-rubber cap removed from the glass cylinder, and the
upper flask tilted until the water begins to flow into the lower one.
142 : BACTERIOLOGY.
The emptying continues by siphon action, and air is drawn in along
the cylinder to replace the water. When the upper flask is empty,
the position of the two is reversed, atid the flow again started.
When a sufficient volume has been drawn through the cylinder, the
outer cap and the cotton-wool plug are replaced, and it is set aside
for the colonies to develop. As an example, twenty-five litres of air
from an open square in Berlin gave rise to three colonies of bacteria.
and sixteen moulds; on the other hand, two litres from a school-
a
Fic. 71.—Hessr’s APPARATUS.
room just vacated by the scholars gave thirty-seven colonies of
bacteria and thirty-three moulds.
Porous substances, such as sand, powdered glass, or sugar, may
be used for the filtration of samples of air; and an apparatus is
employed in a convenient form to be conveyed to the laboratory for
the subsequent examination. ;
Petri’s Apparatus consists of a glass-tube 9 em. long, containing
two sand-filters separated from each other. A known volume of air
is aspirated through the tube. The bacteria are arrested and can
EXAMINATION OF AIR, SOIL, AND WATER. 143
be examined by spreading the sand out in a dish and covering it
with nutrient gelatine ; or it may be shaken up with sterilised water
and plate-cultivations prepared. The sand-filter nearest to the
aspirator should remain free from bacteria.
Sedgwick and Tucker employ a glass cylinder which is drawn out
at one end into a narrow tube to contain sterilised powdered cane
sugar. Both ends of the apparatus are plugged with sterilised
cotton-wool. By means of an exhausting apparatus a known volume
of air is drawn through the tube. The cotton-wool plug is re-
: S , A
Fic. 72.,.-Sep¢wick AND TucKER’s TUBE.
moved, and liquid gelatine is introduced into the cylinder, the
plug is replaced, and the sugar is shaken into and quickly dissolves
in the jelly. The cylinder is then treated in the same way as a
roll-culture, and set aside for the colonies to develop (Fig. 72).
Various forms of “aeroscopes” and “ aeroniscopes” have from
1] = J \
Fic. 73.—PovucuHet’s AEROSCOPE.
time to time been employed. Pouchet’s aeroscope consists of a small
funnel, drawn out to a point below which is a glass slip coated with
144 BACTERIOLOGY.
glycerine. The end of the funnel and the glass slip are enclosed
in an air-tight chamber, from which a small glass tube passes out
and is connected by india-rubber tubing with an aspirator (Fig. 73).
The air passing down the funnel strikes upon the glycerine, which
arrests any solid particles. For a full description of the apparatus
employed by Maddox, Cunningham, and Miquel, reference should
be made to the writings of these authors, and particularly to the
treatise published by the last-named.
SoIL.
Surface soil is exceedingly rich in bacteria. Miquel has com-
puted that there exists in a gramme of soil an average of 750,000
germs at Montsouris, 1,300,000 in the Rue de Rennes, and 2;100,000
in the Rue de Monge. As agents in putrefaction and fermenta-
tion they play a very important réle in the economy of nature ;
but there exist in addition,~badteria in the soil which are patho-
genic in character. Pasteur has succeeded in isolating the bacillus
of anthrax from the earth. Sheep, sojourning upon a plot of
ground where animals with anthrax have been buried, may suecumb
to the disease. Pasteur considered that the spores were conveyed
by worms from buried carcasses to the surface soil. The bacilli
of malignant edema and tetanus are also present in soil. Nicolaier
produced tetanus in mice and rabbits by, inoculating a little garden
earth under the skin.
To obtain a cultivation of the microbes in soil a sample of the
latter must be first dried and then triturated. It may then be
shaken up with distilled water, and from this a drop transferred to
sterilised broth. The employment of solid media is, however,
much more satisfactory: a sample of earth is collected, dried, and
triturated, and a small quantity sprinkled over the surface of ’
nutrient gelatine prepared for a plate-cultivation. In another
method the gelatine is liquefied in a test-tube, the powder added,
and distributed, in the usual way, throughout the medium, which
is then poured out upon a glass plate or made into a roll-culture.
In the same way the dust which settles from the air in houses and
hospitals, or food substances in powder, may be distributed in
nutrient gelatine, and examined both for aerobie and anaerobic
bacteria. The different kinds which develop, must be thoroughly
investigated as regards their morphological and biological charac-
ters, and pathogenic properties.
EXAMINATION OF AIR, SOIL, AND WATER. 145
WatTER.
In the case of water, as in that of air, a knowledge of the
micro-organisms which may be present is not only of interest.
to the bacteriologist, but of the greatest importance in practical
‘ hygiene. Common putrefactive bacteria and vibrios may not be
hurtful in themselves, but they indicate the probability of the
presence of organic matter in which there may be danger. The
detection of Bacillus coli communis may be taken to indicate a
probable contamination with human excreta,
The Microzyme Test which was introduced for the detection
of putrefactive bacteria, consisted in adding three or four drops of
the sample of water to 1 or 2 cc. of Pasteur’s fluid, the nourishing
fluid having been previously boiled in a sterilised test-tube. If the
microzymes or their germs existed in the water, the liquid in a few
days became turbid from the presence of countless bacteria. This
test is of no real value, for it does little more than indicate that
bacteria are present, which we know to be the case in all ordinary
water, and even in ice. On the other hand, the bacteriological test.
of Koch is a most valuable addition to the usual methods of water
analysis. It enables us not only to detect the presence of bacteria,
but to ascertain approximately their number, and to study very
minutely their morphological and biological characteristics. The
-importance of a thorough acquaintance with the life-history of the
individual micro-organisms cannot be too strongly insisted upon.
For example, by such means the spirillum of Asiatic cholera can
be distinguished from most other comma-shaped organisms, and
inasmuch as its presence may be an indication of contamination
with choleraic discharges, such water should be condemned for
drinking purposes, even though we are not yet in a position to
affirm that the microbe is the cause of the disease. The detection
of the bacillus of typhoid fever or of the Bacillus coli communis
in suspected water or milk would be evidence of considerable
importance.
Koch’s test, in short, consists in making plate-cultivations of a
known volume of water, counting the colonies which develop,
isolating the micro-organisms, and studying the characters of each
individual form.
Collection and Transport of Water Samples.—Sternberg’s bulbs,
or Erlenmeyer’s conical flasks of about 100 cc. capacity, may be
employed with advantage for collecting the samples of water. The
latter are cleansed, plugged, and sterilised in the hot-air steriliser.
10
146 BACTERIOLOGY.
When required for use, the plug is removed and held between the
fingers, which must not touch the part which enters the neck of
the flask. About 30 cc. of the water to be examined are intro-
duced into the flask, and the plug must be quickly replaced and
covered with a caoutchouc cap. If collected from a tap, the water
should first be allowed to run for a few minutes, and the sample
should be received into the flask without the neck coming into contact
with the tap. From a reservoir or stream, the flasks may be filled
by employing a sterilised pipette. During transport contact between
the water and cotton-wool plug must be avoided, and if likely to
occur the sample must be collected and forwarded in a Sternberg’s
bulb.
Examination by Plate-cultivation.—The apparatus for plate-
cultivation should be arranged as already described. Crushed ice
Fic. 74.—APpPARATUS FOR EsTIMATING THE NuMBER OF COLONIES IN A
PLATE-CULTIVATION.
may be added to the water in the glass dish to expedite the setting
of the gelatine, so that the plate may be transferred as quickly
as possible to the damp chamber. The caoutchouc cap is removed
from the flask, and the cotton-wool plug singed in the flame to
prevent contamination from adventitious germs on the outside of
the plug. The flask is then held slantingly in the hand, and the
plug twisted out and retained between the fingers. With a
graduated pipette a measured quantity (2; or jy cc.) of the sample is
transferred to a tube of liquefied nutrient gelatine, and the plugs of
the flask and of the tube quickly replaced. If the water is very
impure, it may be necessary to first dilute the sample with sterilised
water. The inoculated tube must be gently inclined backwards and
forwards, and rolled as already explained, to distribute the germs
throughout the gelatine, and the gelatine finally poured on a plate.
When the gelatine has set, the plate is transferred to a damp
chamber, which should be carefully labelled and set aside in a place
EXAMINATION OF AIR, SOIL, AND WATER. . 147
of moderate temperature. In about two or three days the cultivation
may be examined. In some cases the colonies may be counted at
once; more frequently they are so numerous that the plate must
be placed on a dark background, and a special process resorted to.
A glass plate, ruled by horizontal and vertical lines into centimetre
squares, some of which are again subdivided into ninths, is so
arranged on a wooden frame that it can cover the nutrient-gelatine
plate without touching it (Fig. 74). A lens is used to assist in dis-
covering minute colonies. If then the colonies are very numerous,
the number in some small division is counted, if less in some large
one, and an average is obtained from which the number of colonies
on the entire surface is calculated. A separate calculation of the
liquefied colonies should be also made, and their number, as well
as the total number of colonies present in 1 cc. of the sample,
recorded. Any peculiar macroscopical appearances, colours, etc.,
should be noted, and then the microscopical appearances of the
colonies studied. Tastly, examination of the individual organisms
should be made by cover-glass preparations, and by inoculation of
nutrient gelatine, potatoes, and other media.
Instead of plates, Petri’s dishes may be used both for gelatine
and agar-agar cultivations.
i Te ran :
Nis : nara A
ahi 2° @ bs @ & be ® )
5 = o & hf), n° 8 A,
5
Fic. 75.—EsmMArcH’s Rouu-CuLtTure.
at, India-rubber caps; b 6 b, longitudinal line drawn on the tube; ¢,c,c, transverse
lines for counting colonies (FRANKLAND).
Another plan is to take a measured quantity of the sample of
water and prepare a roll-culture, using a large-sized test-tube
(Fig. 75). The colonies can be counted with the aid of a lens (Fig. 76).
Microscopical preparations and sub-cultures can be made from the
colonies, and the anaerobic bacteria can be examined by Frinkel’s
modification of this method (p. 131).
A drop of the sample of water may also be added to liquefied
nutrient gelatine in a test-tube, the organisms distributed, and the
gelatine allowed to solidify in the tube. A rough comparison of
water samples may be made in this way.
Microscopic Examination.—A drop of the water may be mounted
and examined without staining; or allowed to evaporate on a cover-
148 BACTERIOLOGY.
glass, which is then passed through the flame, and stained in the
usual manner.
Parietti’s Method.—As typhoid fever bacilli are apt to be
crowded out by more rapidly growing micro-organisms, some method
had to be devised for restraining the growth of the latter, and
Chantemesse and Widal suggested the use of carbolic acid. Parietti
put this into practice by the method he introduced. This consists in
adding to tubes of broth about five drops of a mixture composed of
sterilised water (100 parts), hydrochloric. acid (4 parts), and carbolic
acid (5 parts). The tubes are first tested by incubation, and are
then ready for use. A few drops of the suspected water are added
Fic. 76.—APPARATUS FOR COUNTING COLONIES IN A ROLL CULTURE.
to the broth, and if it becomes turbid in a day or two the typhoid
fever bacillus is present in the form of a pure-culture.
An excess of bacteria in a fresh sample indicates an excess of
organic matter, and points to possible contamination with sewage.
Where there is such contamination we are very likely to find
pathogenic bacteria; and moreover impure water is a constant
source of danger, for if the contagia of infectious diseases are
introduced they will retain their vitality in such water for a long
period, and will in some cases even multiply, whereas the same
organisms introduced into pure water would in a short time perish.
The actual number of bacteria in water is not of very great
importance, and it must be remembered that if a sample is set aside
for a few days there will be an enormous increase in the number
of bacteria present ; but in dealing with perfectly fresh samples it
EXAMINATION OF AIR, SOIL, AND WATER. 149
may be said that water containing less than 100 bacteria to the
cubic centimetre is very pure water. Water containing 1,000 or
more should be filtered. Water containing 100,000 to 1,000,000
is contaminated with surface water or sewage. It is necessary to
bear in mind that in typhoid fever and Asiatic cholera the excreta
contain the bacteria in great numbers, and wells and streams
receiving surface water may be contaminated in various ways.
The cholera bacillus dies as a rule quickly in distilled water, while
it preserves its vitality for a long time in water of a bad quality.
It is necessary to lay stress upon the fact that a bacterio-
logical analysis may show the presence of pathogenic bacteria when
their detection is not possible by any other means. They may
be present in water in such small numbers that no chemical
analysis would detect any contamination, but as they are living
organisms capable of increasing in a suitable environment, they can
readily be discovered by bacteriological methods.
The examination of rain water, drinking water, tap water, sea
water, various liquids and infusions, by these methods, opens up
a wide field for research. Pettenkofer has shown that impregna-
tion of water containing many bacteria with carbonic acid diminishes
the number of the latter. The examination of waters before and
after filtration, or after addition of chemical substances, are matters
which require further investigation, though a great deal of work has
already been accomplished. The reader will find in Micro-organisms
in Water by P. and G. Frankland, a very complete account of this
subject with valuable analytical tables.
CHAPTER XII.
PHOTOGRAPHY OF BACTERIA.
TueE production of pictures of microscopic objects by photographic
means was attempted at an early date. Some authorities regard
the very earliest recorded experiments as being the first experi-
ments alike in photography and micro-photography. The experi-
ments of Wedgwood and Sir Humphry Davy were embodied in
a paper read before the Royal Institution in 1802. They obtained
with the solar microscope impressions upon paper, and with greater
success upon white leather, though the results were transitory when
exposed to daylight.
In 1816 Nicéphore Niepce described his experiments in con-
nection with fixing the image obtained by the camera. He was
at first only able to obtain negatives, and these were transitory.
But, after joining with Daguerre, who had been experimenting
in the same direction, a process was invented which was published
in 1839 under the name of daguerreotype.
This invention, and the rapid improvements which followed,
were taken advantage of by Reade, Donné, Hodgson, Kingsley, and
Talbot, who were early workers in the field of micro-photography.
So early as 1845 it is stated that Donné produced a work
illustrated with engravings copied from daguerreotypes.
Subsequently this interesting branch of photography was taken
up by many in France and Germany, in America, and in England.
Of those to whom we are indebted for the literature of the subject,
and for many improvements, the names of Wenham, Dancer,
Draper, Maddox, Shadbolt, Redmayne, Woodward, Highley, Deecke,
Moitessier, Gerlach, Koch, Sternberg, Frankel, Pfeiffer, and Pringle
may especially be mentioned.
Of these workers the name of Woodward stands pre-eminently
foremost. His skill in microscopical manipulations, combined
with access to the very best apparatus and objectives, placed at
150
PHOTOGRAPHY OF BACTERIA. 151
his disposal in the Museum at Washington, enabled him to obtain
photographs of diatoms which probably have never been surpassed.
To Koch belongs the credit of being the first to extend the
application of micro-photography to the delineation of bacteria.
A series of instructive photographs was first published by him
in 1877. These were photographs of cover-glass preparations,
and all admirably illustrated the subjects from which they were
taken; while two, showing the flagella of bacilli and spirilla,
were triumphs in this new departure.
Lewis, in India, was one of the first to illustrate his writings on
the subject of micro-organisms by means of photographs.
About the same time Sternberg, in America, took some excellent
photographs of bacteria. Heliotype reproductions of these were
published in 1884.
Hauser and Van Ermengem and many other bacteriologists
successfully resorted to photography for illustrating their researches,
and Frinkei and Pfeiffer’s, and Itzerott and Niemann’s atlases of
photographs of bacteria, in microscopical specimens and cultivations,
are especially worthy of mention.
Opinions have differed widely as to the merits of photographic
illustrations. Many, taking the standpoint solely of a comparison
with drawings, have decried their use. By judging from such a
comparison alone the real value of photographs may be lost sight
of. On the other hand, many who have looked at the question
from all sides, have been led to value even a defective photograph
more than an ordinary drawing.
In his first publication on this subject, Koch strongly advocated
photography on the ground that illustrations would then be as true
to nature as possible. The photographs which accompanied his
paper were all taken from preparations of bacteria which had
been made from blood, cultivations, or infusions, by drying a
thin layer on a cover-glass and staining, or from specimens prepared
in the same way but left unstained. But when, having committed
himself to this opinion, Koch attempted, later, to photograph the
bacteria in animal tissues, he was led to modify his previous
conclusion. For though no trouble was spared, yet disappointing
results were met with. This was owing, he explains, to the fact
that the smallest and most interesting bacteria can only be made
visible in animal tissues by staining them, and thus obtaining the
advantage of colour.
This introduced the same difficulties which are met with in
photographing coloured objects, such as tapestry and oil paintings.
152 BACTERIOLOGY.
As these difficulties had been to a certain extent obviated by the
use of eosin-collodion, Koch adopted the same method for photo-
graphing stained bacteria. By the use of eosin-collodion, and by
shutting off portions of the spectrum by coloured glasses, he
succeeded in obtaining photographs of bacteria which had been
stained with blue and red aniline dyes. But, owing to the long
exposure which was necessary, and the unavoidable vibrations of
the apparatus, the results were so wanting in definition that they
not only proved unsatisfactory as substitutes for drawings, but did
not in some cases give any evidence of what was to be seen in the
preparations.
Koch, in consequence, stated that he would abstain from
publishing photographic illustrations until he had the advantage
of improved methods.
We find, however, in spite of this, that in 1881 Koch published
a series of reproductions from his negatives in illustration of what
could be accomplished by photography.
Here again we find that many of the photographs of cover-
glass preparations were admirable, but those of tissue-sections gave
evidence of the difficulties Koch encountered, and were undoubtedly
unsatisfactory from the want of flatness of field, some of the
ilustrations recalling rather a map of a mountainous country than
a microscopical preparation. ,
In consequence of the difficulties met with in attempting to
photograph bacteria stained with the aniline dyes most commonly
used, Koch recommended that the preparations should be stained
brown, pointing out as his reason that, though the bright and
concentrated colour of the red and blue aniline dyes catches the
eye far more readily than the somewhat sombre brown colours,
yet no one up to the time of his publication had succeeded in
obtaming good photographs of bacteria which had been stained
either blue or red, and mounted in Canada balsam, while there was
no difficulty in obtaining photographic representations of prepara-
tions stained yellow or brown.
Though this stain could be easily employed in most cover-glass
preparations, it was by no means easy to obtain a good differential
stain of bacteria in the tissues by employing Bismarck brown.
An attempt was, therefore, made to photograph preparations
stained blue and red by the aid of the dry-plate process, and by
interposing glasses of suitable tints. After many fruitless experi-
ments this method had to be abandoned, and the method of staining
the object brown was adopted. In many cases this gave excellent
PHOTOGRAPHY OF BACTERIA. : 153
results; in others again, compared with the results of staining
with blue or red stains, there was much to be desired, and further
improvement was needed.
That a stain, such as yellow or brown, must be employed which
absorbs the blue rays, and acts on the sensitive plate like black,
which absorbs all the light, constituted the first condition laid
down by Koch as an essential for success. It was further pointed
out that the suitability of the stain could be ascertained by first
passing the light, to illuminate the preparation, through a solution
of ammonio-sulphate of copper, under which condition the bacteria
would appear black on a blue ground.
The second condition was, that sunlight must be employed, but
that direct projection upon the object was disadvantageous, and it
must, therefore, be diffused by the interposition of one or more plates
of ground glass.
Lastly, an illuminating condenser was recommended, of such
construction that the diffused sunlight brightly illuminates the object
from all sides.
Sternberg encountered the same difficulty in photographing red,
blue or violet preparations, while he produced excellent pictures of
preparations stained with aniline brown, or a weak solution of iodine
(iodine grs. iij, potassic iodide grs. v, distilled water grs. 200). Thus
the results of a large number of attempts to photograph the tubercle
bacillus in sputum, only ended in producing such extremely faint
impressions, that any one unacquainted with the object as seen under
the microscope could form scarcely any idea of its form or minute
structure with even an accompanying explanation and the closest
inspection of the photograph.
Dufrenne, in attempting to photograph the same object by the
ordinary method, found the plates were uniformly acted on, or the
image was so faint, or so lacking in contrast, that they were useless
for obtaining proofs on paper or glass. By interposing green glass
between the objective and the sensitive plate, so that the red rays
were absorbed, while the green rays passed through and acted on
the plate, he states that better results were obtained.
The work of Hauser illustrated the great value of photography
in the production of pictures of impression-preparations and colonies
in nutrient gelatine. To give the general effect, as well as faithfully
reproduce the minute details in these difficult subjects would in most
eases create insurmountable difficulties, except to the most accom-
plished draughtsman.
Hauser employed Gerlach’s apparatus and Schleussner’s dry
154 BACTERIOLOGY.
plates, and obtained the illumination by means of a small incan-
descent lamp, which gave a strong, white light, with three or
four Bunsen elements. In another respect Hauser’s results were
of practical value. The preparations to be photographed were
stained brown as recommended by Koch, but they were mounted in
the ordinary way in Canada balsam. The objection to the mounting
medium most commonly employed was thus set aside. The prevalent
idea, however, that the preparations must be stained brown was still
a formidable obstacle, and the way out of this difficulty was clearly
shown by Van Ermengem’s photographs. These were pictures of
comma-bacilli which had been stained with fuchsine and methyl
violet. These photographs afforded the first practical illustration
of the value of isochromatic plates in micro-photography which had
been previously noted by Van Ermengem in 1884, and their intro-
duction marks a distinct era in the progress of micro-photography.
A short explanation may be given of what is meant by isochro-
matic, or what have been more properly termed orthochromatic dry
plates. The difficulties encountered in photographing certain stained
preparations have been mentioned. It is a familiar fact that in
portraits, blue or violet comes out almost or quite white, while
other colours, such as yellow, are represented by a sombre shade
or perhaps black. This failure in correctly translating colours is
explained by the want of equality between the strength of the.
chemical and luminous rays. If the rays of the spectrum are pro-
jected upon a photographically sensitive surface, the greatest effect
is found to take place at the violet end. In other words, the violet
and blue rays are more actinic or chemically powerful, while the
yellow and orange have scarcely any effect. The dyes employed in
staining give corresponding results: blue and violet give but a faint
impression, yellow and orange a black picture. These results are
most clearly demonstrated in a photograph of an oil painting taken
in the ordinary way ; and they led to experiments being made which
have resulted in orthochromatic photography.
The effect of interposing coloured glasses has already been
referred to. It was found later that, if plates were coloured yellow,
e.g., with turmeric, the blue and violet rays were intercepted, and
their actinism reduced. In 1881, Tailfer and Clayton produced the
so-called isochromatic plates. The emulsion of bromide of silver
and gelatine was stained with eosin, and it was claimed that colours
would be represented with their true relative intensity. Chlorophyll
and other stains have been tried, and by such methods the ordinary
gelatine dry plates can be so treated that they will reproduce
latte gm
PHOTOGRAPHY OF BACTERIA. 155
various colours, according to their relative light intensity, and thus
be rendered iso- or, what is now more commonly known as, ortho-
chromatic.
APPARATUS AND MATERIAL.
Micro-photographic Apparatus.—As is well known, various forms
of apparatus have from time to time been recommended and em-
ployed by different workers.
Many use the microscope in a vertical position, with the camera
superposed or fitted to the eye-piece end of the microscope tube;
or the microscope tube may be screwed off from the body of the
microscope, and a pyramidal camera adjusted in its place, the base of
the pyramid being represented by the ground glass screen.
Others again prefer that the microscope and camera should be
arranged horizontally.
In another form the ordinary microscope is dispensed with, the
objective, stage, and mirror are adapted to the front of the camera,
and provided with suitable arrangements for holding the object,
supporting the mirror, and adjusting the different parts.
Lastly, the camera may be dispensed with, the operating-room,
which must be rendered impervious to light, taking its place, while
the image is projected and focussed upon a ground glass screen,
which has a separate support.*
The horizontal position affords greater stability than the vertical,
so that the former is to be preferred. The vertical model with the
camera fixed to the microscope is particularly to be avoided, as the
weight of the camera bears directly upon the microscope, and must
affect the fine adjustment, and any vibration in one part of the
apparatus is communicated throughout.
The simplest apparatus consists of a camera fixed upon a base-
board four or five feet in length, upon which the microscope can be
clamped, and which carries also a lamp and a bull’s-eye condenser
(Fig. 77).
Simplicity and economy must always be borne in mind in
recommending any apparatus of this kind, for to insist upon the
“necessity of a very elaborate apparatus, or a specially fitted-up room,
or that a special room should be built with windows facing in a
definite direction, will in most cases at once place photography beyond
the reach of those who might otherwise employ it. Yet to fulfil
* For an excellent account of the forms of apparatus which have been
employed by different workers the reader is referred to the section on Micro-
photography in Beale’s How to work with the Microscope.
156 BACTERIOLOGY.
all the purposes for which the apparatus may be required, including
the employment of the highest powers, and also that one may be
enabled to work for long intervals of time with due comfort, an
accurate and complete apparatus will be found to be most desirable.
Though most preparations will admit of being photographed
when the stage of the microscope is vertical, yet if we require to
photograph micro-organisms in liquids, or colonies upon partially
liquefied gelatine, the apparatus must admit of being placed so that
the stage of the microscope becomes horizontal. In addition, the
apparatus is rendered somewhat complex if we employ powerful
TTT
IMCS
Fic. 77.—HorizontaL Mick0-PHOTOGRAPHIC APPARATUS.
artificial light. Sunlight, no doubt, is the best and cheapest, but
it is not always available, especially in a city like London; and,
moreover, evenings and dull days will probably be the very time
which can be best spared for this work. We must, therefore, fall
back upon the paraffine lamp, or the magnesium, oxyhydrogen, or
electric light.
To fulfil all these conditions Swift has constructed an apparatus
under the author’s directions (Fig. 78). It is merely a modification
of the ordinary horizontal model, which admits of being readily placed
PHOTOGRAPHY OF BACTERIA. 157
in the vertical position, while the illumination is supplied from an
oxyhydrogen lantern.
To place the apparatus in the vertical position two small hinged
‘SALVUVddW OlHAVUDOLOHA-OWOIP AIAISUAATY—'S) “OLA
brackets, at the end distant from the camera, are forced up with
a smart blow of the hand. The corresponding ends of the stretcher
bars are dislodged from their fittings, and allowed to descend ; when
158 BACTERIOLOGY.
horizontal, the opposite extremities of the bars are easily released
from their sockets. The leg or support at this end can then be
Fie. 79,—REVERSIBLE Mick0-PHOTOGRAPHIC APPARATUS ARRANGED IN THE
VERTICAL POSITION.
turned up and fixed underneath the apparatus by a button, and
the end of the apparatus itself gently lowered to the ground.
PHOTOGRAPHY OF BACTERIA. 159
A hinged end-piece is also to be turned out to increase the base upon
which the whole apparatus will stand when raised to the vertical.
The two-legged support at the opposite end of the apparatus is
next worked down by a quick thread screw, and on raising the
apparatus to the vertical, the two-legged support drops to the
ground, and assists in maintaining the stability of the whole. If
it is thought necessary, a simple means can be readily devised for
clamping the apparatus, in either position, to the wall of the room,
so as to eliminate as much as possible all chances of vibration. A
second quick thread screw moves the base-board upon which the
_camera and central sliding-board are mounted, so that the camera,
muicroscope and lantern can be raised to a convenient height from
the ground.
The various parts of this apparatus. may be described in
detail.
The Microscope and its Attachments.—It is most essential that
the microscope should be perfectly steady. The microscope was made
by Zeiss, and to ensure steadiness, the horse-shoe footpiece fits under
a projecting ledge, and is then clamped by a cross-piece, so that
it is firmly fixed.
The microscope with the means for clamping it and the oxy-
hydrogen lantern are carried upon an independent sliding-board,
which admits of movement to or from the camera. Thesliding-board
also moves upon a centre, which enables the microscope to be turned
out from the median line; in fact, to be turned at aright angle to the
position it occupies when ready for the exposure. The object of this
contrivance is to enable the operator to sit down by the side of the
apparatus, and with comfort to arrange the object in the field of the
microscope. On turning the microscope back into the median line, it
is fixed in the optical axis of the apparatus by means of a suitable
stop. The sliding-board is provided with a small grooved wheel
receiving an endless cord, made of silk or fishing-line, which passes
round the grooved, milled head of the fine adjustment of the
microscope. When the: slidmg-board is returned to the median
line of the apparatus, the milled wheel connected with the fine
adjustment impinges upon the wheel of the long focussing rod.
The latter is provided with an india-rubber tire, which grips the
teeth of the milled wheel, and thus the long focussing rod is placed
in connection with the fine adjustment of the microscope.
Illumination.—The oxy-hydrogen lamp has been more frequently
employed by the author than the paraffine lamp, partly on account
of the diminished time in exposure, especially when employing very
BACTERIOLOGY.
160
PHOTOGRAPHY OF BACTERIA. 161
high powers ; this is of great importance where there is likely to be
vibration from passing traffic. With rapid plates and the highest
powers, the exposure has only been two or three seconds, whereas
with the paraffine lamp it may vary from three to ten minutes, or
even longer.
Walmsley gives the following table for exposures with the
paraffine lamp :—
14 inch objective 3 to 45 seconds.
aa - € 5-90e 3
aa» is 3 ,, 3 minutes.
Beg 3 2 55 TF ;
to ” ” 4 ” 10 ”
The illuminating apparatus represented in the accompanying
engraving (Fig. 78) consists of a lantern which not only moves
together with the microscope on the central sliding-board, but
can be moved independently to or from the microscope, and be
clamped with screws at the requisite distance for obtaining the best
illumination. It is provided with two 3-inch condensing lenses of
long focus, constructed of optical glass, which is much whiter than
that used for ordinary lantern condensers. The lime-cylinders should
be of the hardest and best quality, as they give a more actinic light
than those made of soft lime. The “ Excelsior” lime-cylinders are
strongly recommended. They are.supplied in hermetically sealed
tins which can be easily opened and re-sealed, so that a cylinder can
be taken out and used, and the rest preserved for a future occasion.
The hydrogen can be obtained by using the coal-gas supplied to the
house, and the oxygen should be supplied preferably in a compressed
state in iron bottles. Not only are the bottles much less cumbrous
than the bags, but a small quantity of gas can be used, and the
residue left for an indefinite time; moreover, the gas is always
at hand to be turned on when required. On the other hand, the
retention of unused gas in bags is liable to cause their corrosion,
owing, it is belieyed, to impurities which are carried over in the
manufacture of the oxygen. If gas is not laid on in the house, then
it also must be procured in a compressed state in bottles. As the
blow-over jet is recommended on account of its safety, the bottles
should be supplied in this case with a supplementary valve. It is
then just as easy and free from danger to employ the compressed
gas as it is to make use of the house-supply.
The Camera.—A. long-focus, half-plate camera is mounted upon
a sliding platform. This admits of the camera being pushed up to
ll
162 BACTERIOLOGY.
the microscope when it is in the long axis of the apparatus, so as to
make a light-tight combination, The opening which is filled in an
ordinary camera by the lens can be shut off by means of an internal
shutter, which is opened and closed by turning a screw at the side
of the camera. The dark-back is provided with plate-carriers, so
that either half, quarter, or lantern-size plates can be employed. It
will be found convenient to have two or more dark-backs, so that
several plates may be exposed without rearranging the light for
each exposure,
Much more elaborate and expensive micro-photographic cameras
have been constructed by Zeiss, and also by Swift. The latter has
Fic. 81.—PHoToGRAPH Or AN IMPRESSION PREPARATION.
carried out a suggestion made by Pringle for a support at the
ocular end (Fig. 80).
The Dark-room.—In every bacteriological laboratory there should
be a developing room provided with shelves, gas, water-tap, and sink,
but these arrangements are not absolutely indispensable. All that
is essential is a room impervious to light; and a closet or cupboard,
if it can be ventilated, will answer perfectly well, with a jug and
basin instead of the tap and sink. The steam-steriliser employed
in the preparation of nutrient media for cultivating bacteria, if not
required at the time for such purposes, may be filled to the brim
with water, and will form an excellent cistern and tap, while a pail,
or small sanitary bin, may be utilised as a sink.
Various kinds of lamps are made for the dark-room, burning
PHOTOGRAPHY OF BACTERIA. 163
either candles, oil, or gas. In any case, the light must pass through
two thicknesses of ruby glass.
Dry Plates—A small supply of any of the ordinary plates in
the market may be procured for preliminary trials in acquiring a
knowledge of the processes ; but to overcome the difficulties of certain
stained preparations, the isochromatic or orthochromatic plates should
be used. The 7 plate will be found to be the most suitable size.
There are numerous formule for the requisite solutions for
developing and fixing the negatives, and instructions are usually
enclosed. in the boxes of dry plates, but it is best to abstain from
trying a number of different formule, as it leads to a great expendi-
ture of time. There is a temptation to do this, it being supposed
that there is probably some great advantage in one formula over
another. It is much better to get accustomed to the behaviour
under different exposures of one, or perhaps two methods.
In France the iron developer is much in vogue, and is recom-
mended by Tailfer and Clayton for use with their isochromatic
plates. It has the advantage of great simplicity in the mode of
employment, and, therefore, is very suitable for a beginner. In
England, on the other hand, the alkaline developer is very
much used, as it gives more command over the plate, enabling the
photographer more fully to compensate for incorrect exposure.
It is very desirable before attempting to take photographs with
the microscope to learn how to take photographs with an ordinary
landscape camera, and to get thoroughly accustomed to the use of
some good developer, so that mistakes may be corrected and the
clearest and sharpest negatives obtained.
PracticaL ManipuLation.
Arrangement of Apparatus.—For working with the paraffine
lamp, the mode of procedure is, as regards the illumination, briefly
as follows. The sub-stage condenser is dispensed with when a
low power is employed, as well as the mirror, and the lamp is
so placed that the image of the flat of the flame appears accurately
in the centre of the field of the microscope. A bull’s-eye condenser
is then interposed, so that the image of the flame disappears, and
the whole field is equally illuminated. With high powers the
sub-stage achromatic condenser is necessary, and a more intense
illumination is obtained by using the flame edgewise. In using a
low power with the oxyhydrogen light, the lantern is withdrawn
some little distance from the microscope, and the top combination
of the achromatic condenser removed.
164 BACTERIOLOGY.
It is best to begin with the use of a low power, and a trial
object, such as the blow-fly’s tongue, spine of Echinus, or trachea
of silkworm.
In order to explain the management of the apparatus (as
represented in Fig. 78) the steps in the arrangement of the
apparatus and exposure of the plate will be described in detail
for the employment of a high power and the oxyhydrogen light.
The solutions being ready for use, it is proposed to take a photograph
of tubercle bacilli in sputum, with a ~, apochromatic oil-immersion
objective. The first point to claim attention is the arrangement
“of the light. Having lighted the gas at the hydrogen jet, the
lime-cylinder should be revolved until heated equally all round.
The oxygen is then carefully turned on until only a small spot
of incandescence is produced. The central sliding-board is turned
out, a low power screwed on to the microscope, and the image of
the bright spot focussed and accurately centered. To protect the
sight, an eye-piece provided with a smoked glass shade is used.
The immersion objective is then substituted for the low power,
and the oxygen turned on until the right admixture of gas is
obtained to produce a brilliant illumination. It is well at this
stage to sit down to focus the selected object, and to spend some
little time in searching for the most characteristic part of the
specimen to be photographed. This being decided upon, the eye-
piece is carefully withdrawn, and the central sliding-board rotated
back into the median line. To make a light-tight connection
between the camera and the microscope, the camera is pushed up
until a velvet-lined tube, which occupies the position of the lenses
of an ordinary camera, is enclosed within a short wide tube which
is adapted to the eye-piec2 end of the microscope.
On opening the camera-shutter the image will be projected upon
the ground glass screen of the camera. It is necessary, however,
to obtain the exact focus, and to effect this the ground-glass
screen is turned away, and the dark-back with a piece of plain
glass is substituted. Here again time may be well spent in
getting the sharpest image, with the aid of a focussing glass of
proper focal length.
The greatest delicacy in manipulation is necessary, as in working
with such high powers a turn too much of the fine adjustment will
cause the image to vanish. Having determined the best visual
focus, which will be found with the high-power objectives of most
makers to correspond with the chemical focus, the dark-back must
be cautiously removed, to prevent any vibration, and the plain
PHOTOGRAPHY OF BACTERIA. 165
glass replaced by a sensitive plate. To effect this change, the
operator retires to the dark-room, and opens a box of plates with
as little exposure to the red light as possible. Having removed
a plate, it is necessary to ascertain which is the sensitive side.
This may be done by momentarily exposing it to the red light,
and seeing which is the sensitive side by the dull appearance of
the film. A less satisfactory way is to moisten the tip of a
finger, and press it at one corner of the plate. The film side will
be recognised by imparting a sticky sensation. The film must be
dusted with a camel’s-hair brush, as well as the dark-back, and
the plate is placed film-side downwards in the dark-back, which
is then securely closed.
Care should be taken that the plates then remaining in the box
are packed away before light is admitted to the dark-room.
Exposure of the Plate-—On returning to the apparatus, the
camera-shutter is closed. Then the dark-back is gently slid into
its place, and its slide withdrawn. A few moments are allowed
to elapse, so that the least possible vibration, which might be
caused by inserting the dark-back, has had time to cease, and all
is ready for the exposure.
In the case of the object we have selected, three seconds will
probably be the exposure required. ‘This is done by opening and
closing the camera-shutter with one hand, while a watch can be
held in the other. The slide of the dark-back is then carefully
closed, and the plate is ready to be carried off to the developing
room. :
As the light will not be again required until the next exposure,
the oxygen must be turned off, while the coal-gas may be allowed
to play over the lime.
Development and Fixation of the Image.—It is well to be
systematic, and therefore, before the plate is taken out of the
dark-back, light is admitted to the dark-room, and everything
arranged so that the position of the trays and bottles may be
remembered in the dark. First, let the ruby lamp be lit, place
two dishes or trays close by, and a row of four dishes within easy
reach. Pour out some fixing solution in the first porcelain dish,
alum in No. 2, and water in Nos. 3 and 4, Put the necessary
quantity of “pyro” solution into the glass measure, and place
it with the ammonia drop-bottle in front of the ruby light.
Then, when all light except that from the ruby lantern has been
excluded, everything is ready to commence the development of the
plate.
\
166 BACTERIOLOGY.
Opening the dark-back, the plate may be turned out on to the
palm of the hand. The film side is then uppermost, and the plate is
to be transferred in the same position to a tray, and covered with
water. This is to soak the film and obtain an equal action of
the developer ; or the solution of fresh pyro may be poured on to
the plate without previous soaking, if the flow is uniform, and the
formation of bubbles avoided. In the first case the water is run off
and the pyro allowed to flow evenly over the plate. To protect the
plate from prolonged exposure to the ruby light, a second tray may
be inverted over it, or the developing tray covered with a piece
of card-board. Gently rock the tray for a minute or so, then
to a few drops of ammonia in a measuring glass add the pyro
from the developing tray, and pour the mixture back again
over the plate. After again gently rocking the tray for a few
minutes, more ammonia is added by drops in the same way.
If the exposure has been properly timed,—and the time necessary
must be ascertained by trial for each preparation,—the image will
gradually begin to appear, and the action must be allowed to
continue until sufficient density has been obtained. To determine
this requires some experience. It is generally recommended to take
the plate out of the tray and hold it for a moment, film-side towards
the operator, in front of the ruby light. Though the plate is not
nearly so sensitive when the image has commenced to develop, and
there is, therefore, not the same danger of fogging, a safer plan is
to occasionally turn the plate film downwards in the tray, and when
the image appears on the back the development will be found to be
completed.
With such a preparation as tubercle bacilli in sputum it is
not easy to trace the gradual formation of the image, and hence the
advantage of commencing with a well-marked object such as the
blow-fly’s tongue. It is then easy to watch the gradual progress of
the image. The bright parts or high-lights appear first, then
gradually the half-tones, or less brightly-lighted parts, and lastly
every shade except the deepest shadows is represented. When,
however, all action seems to have ceased, we must still wait until
we have judged, in the manner already described, that the density
is sufficient. This being determined, we pour off the developing
solution and thoroughly wash the plate with water. It is then
ready to be placed in dish No. 1, containing “hypo,” and here it must
be left for some minutes after all appearance of creaminess has
disappeared from the back. White light may now be admitted, the
plate removed from the hypo and thoroughly washed under the tap,
PHOTOGRAPHY OF BACTERIA. 167
and then placed in dish No. 2. When another plate is ready to
take its place, transfer it to dish No. 3, and then to No. 4, and,
after a good final washing under the tap, place it upon a rack
to dry. If there is any tendency for the film to detach itself from
the plate, ‘to frill,” the alum bath must be used before fixing, as
well as after.
Frilling or blistering may be due to an error in manufacture,
and is liable to occur in hot weather, or when using a developer too
strong in ammonia. If it occurs during washing or fixing, the alum
bath must be employed before the hypo. ogging, or the appear-
ance of a veil over the plate, may arise from error in the manu-
facture, from admission of extraneous light, from over-exposure,
or from prolonged exposure to the ruby light during development.
Care must be taken that the camera and dark-room are light-tight.
Crystalisation, or powdery deposit, upon the negative when dry, is
due to insufficient washing out of the hyposulphite of soda. Thin-
ness of the image, or want of density, may be due to insufficient
development, too weak a developer, or too short or too long an
exposure. Too great density results from too long immersion in the
developer.
Spots may sometimes occur upon the negatives. They may be
caused by dust upon the plate or by air bubbles in the developer.
In the text-books of photography full accounts of failures will be
found, their causes and prevention; but it will be advantageous
when these difficulties are encountered to take the negatives to a
skilled photographer and get advice upon them. It is necessary
to persevere, and not be disheartened if several negatives have to
be made of a preparation before a successful result is obtained.
It may here be remarked that the beginner will far more
rapidly learn the technique if he avail himself of a practical demon-
stration from a photographer. When he has learnt to obtain suc-
cessful negatives, if he prefer silver prints, and time is an object, it
will be found to be true economy to get the printing and mounting
done by a professional photographer. The credit of a successful
photograph of bacteria is due to the bacteriologist who prepares
the microscopical specimen and obtains the negative.
Determination of the Amplification.—The amplification varies
not only with the objective employed, but with the distance of
the focussing screen from the object. In order to ascertain the
amplification afforded by a certain objective at a certain distance, a
photograph should be taken, under the same conditions, of the lines of
a micrometer slide. It is easy then to calculate the amplification
168 BACTERIOLOGY.
obtained in the micro-photograph ; supposing, for example, in
the micro-photograph the lines which are 759 inch apart are
delineated 1 inch apart, the magnifying power must be 1,000
diameters. Moreover, having thus ascertained the amplification,
we can accurately compute from the photograph the size of the
objects taken.
Value of Photographs.—It is not necessary to compare the
relative merits of diagrams and photographs. Diagrams which do
not purport to be accurate representations, but are
intentionally the means for simplifying instruction,
‘4 will always be valuable, even if we have the
original preparations under the microscope before
cle us. We must consider the relative merits of
photographs and of drawings which purport to be
exact representations of what is seen under the
microscope. Thus in the case of micro-organisms,
when their biological characters are studied under
low powers of the microscope, photographs are
preferable because they give a more faithful re-
presentation. At the same time, apart from this
comparative value, we must not lose sight of the
actual value of photography in placing within the
reach of the student or investigator, who may not
be a draughtsman, a most valuable means for
illustrating all kinds of preparations.
For double-stained or triple-stained tissue pre-
parations an accurately coloured drawing leaves
Fic. 82. — Puoto-
GRAPH OF A ; ‘ s
Currivation or little to be desired; but if we reproduce the same
Bacittus = AN- by a wood engraving, and so lose the advantage
kr ics of the coloured picture, which is instructive in
indicating the method of staining, a photograph will, in many cases,
be far more satisfactory.
When we have to deal with the growth of bacteria en masse,
as in test-tube and plate-cultivations, with colonies as seen under
a low power of the microscope, and with impression-prepara-
tions both under low and high powers, unless the bacteriologist
is a most accomplished draughtsman as well as an accurate and
reliable observer, photography undoubtedly affords the best mode
of illustration. The apparatus being ready and at hand, a negative
can be produced in a few minutes of a preparation which, from the
amount of detail it contains, would take perhaps several hours to
draw and colour. From that negative any number of facsimiles can
PHOTOGRAPHY OF BACTERIA. 169
be obtained, whereas an original drawing, even in the best hands,
if cut on wood or lithographed, is almost certain to fall short of
being an exact copy.
With regard to individual bacteria, the result is more satisfactory
in many cases than a drawing; for there is the advantage of being
absolutely certain that any particular structure, form, or shape
which may be represented is actually what exists, and not what
may have been evolved by unconscious bias in the mind of the
observer. Many illustrations might be given of this. Thus Lewis,
who was a most conscientious observer, published an account of
organisms in the blood of rats in India, and illustrated it with a
wood engraving and with micro-photographs. The identity of the
organisms which were found in the common brown rat of this
country was established much more readily from these photographs
than from the wood engraving or the description in the letterpress.
A micro-organism, even under the highest powers, appears as
so minute an object that to represent it in a drawing requires a
very delicate touch, and it is only too easy to make a picture which
gives an erroneous impression to those who have not seen the
original. If, on the other hand, to represent the object more
clearly we draw an enlarged picture, we can only do so by repre-
senting what we think the object would be like if it could be
amplified to the size represented. In such cases a photographic
enlargement is certainly more valuable.
Photography enables us also to record rapid changes, and it
is possible that as the art advances we may find that the film is
more sensitive than the human retina, and brings out details in
bacteria which would be otherwise unseen.
Photographs can be readily transmitted by post, and when we
can neither make a great number of preparations to illustrate
some object, nor perhaps be able to go to the expense of having a
drawing reproduced, this method will be of value in enabling others
to benefit by our observations.
The author is convinced that if the employment of photography
is encouraged in bacteriological and other research laboratories for
depicting microscopical preparations and cultivations of bacteria, the
results of increasing experience and practice will lead to its being
made more general use of as a faithful and graphic method, valuable
alike for class demonstrations and for illustrating publications.
PART ILI.
ETIOLOGY AND PREVENTION OF INFECTIVE
DISEASES.
171
CHAPTER XIII.
SUPPURATION, PYAMIA, SEPTICEMIA, ERYSIPELAS.
ABSCESS.
WHEN inflammation is followed by an accumulation of leucocytes
and of plasma which does not coagulate, the result is a white or
creamy liquid called pus, and when the surrounding tissues are
involved so that a cavity develops containing pus, we have what
is termed an abscess. The almost constant association of bacteria
with the production of pus has created a belief that they are
the direct cause of suppuration. Ogston found micrococci present
in all acute abscesses, and concluded that acute inflammation was
invariably due to their presence. The fact that inflammation
occurs more frequently in the external tissues of the body is
accordingly explained by the ready entrance of bacteria which
are in the air; and suppuration following pericarditis, pleurisy,
and other conditions in which air is excluded is attributed to the
presence of pyogenic cocci, which have gained access by the blood
stream. There is no pyogenic organism constantly present, but
several different species of bacteria have been isolated from pus
and carefully studied, and the antiseptic treatment is based upon the
principle of excluding bacteria in surgical operations, and destroying
any which may have previously obtained access to wounds and
broken surfaces. Inflamed tissue and pus form a most suitable
medium for the growth of bacteria, which in some cases are
unquestionably only accidental epiphytes,
In tuberculosis, actinomycosis, and glanders, pus formation may
take place without the presence of pyogenic cocci; and it is generally
believed that chemical irritants, such as croton oil, turpentine, iodine,
cadaverin, and tuberculin, will excite the formation of pus in the
absence of bacteria, although Klemperer, after a number of very
careful experiments, maintains that no genuine pus will be produced
if the chemical irritants are first carefully sterilised.
173
‘
174 INFECTIVE DISEASES.
The bacteria which have been isolated from pus include :—
Staphylococcus pyogenes aureus, albus, and citreus, Staphylo-
coccus cereus flavus and albus, Streptococcus pyogenes, Micrococcus
pyogenes tenuis, Micrococcus pneumonie croupose, Bacillus pyo-
cyaneus, Bacillus pyogenes fcetidus, Micrococcus tetragenus, Bacillus
intracellularis meningitidis, Gonococcus, Bacillus septicus vesice,
Urobacillus liquefaciens septicus, Bacillus typhosus, Bacillus coli
communis, Bacillus anthracis, Bacillus tuberculosis, Bacillus mallei,
and Actinomyces.
Fic. 83.—SuPPURATION OF SUBCUTANEOUS TISSUE.
d, Leucocyte containing micrococci; d’, leucocyte with pale nucleus showing
necrosis; e, fixed connective tissue cells, much enlarged, containing several
nuclei, of which some (n’) are pale and necrotic ; numerous cocci, diplococci,
and short chains. (CoRNIL and RANVIER.)
Some idea of the distribution of the bacteria most commonly
occurring in pus may be gathered from the records made by Passet
and by Karlinski.
Passet examined acute abscesses, and found Staphylococcus
pyogenes aureus and albus in 11 cases, Staphylococcus pyogenes
albus alone in 4, Staphylococcus pyogenes albus and citreus in 2,
Streptococcus pyogenes alone in 8, Staphylococcus pyogenes albus
and Streptococcus pyogenes in 1, and Staphylococcus pyogenes albus
and citreus, and Streptococcus pyogenes in 1.
SUPPURATION, PYZMIA, SEPTICEMIA, ERYSIPELAS. 175
Karlinski tabulated his cases thus :—
a} a , g lta
gee |s2l2_iea| 2/43! &
82/88/95 198/35/38|s5| 8
g | 84) 85) 8<|88 38 |ke|/38| &
DISEASE. & |B8|b8 |58 g 82/83) os a
ae la8 (a8 | s5/88 (|ae/s2| 2
SP | sel so/ sh lee | wee | B
ae | a = na ee & |e = é
Mastitis . . 36 | 22] 4 4) 6) eyes |e
Subcutaneous Abscess 30110] 2 8 6] 2 7 ee oe
Phlegmon . i .| 24) —]—)]—] 24] —] —] - _
Furuncle ‘i 90° | 6) = | TO ea) a ee
Bubo . ‘ Pe (is a ae a El a ee a ee
Subperiosteal Abscess TO Ge) aes Oe i ee ee ee ee |
Panaritium Cutaneum 16/ 7)—| S|) l— | =
Panaritium Osseum : 100) 27 ee) OB eee | ee Se S|
Dental Abscess . 10; 1/—) 4/ 1({ 38 A 4)
Hordeolum F “40: | @ |= | ets pease || | 8h eee
Abscess of the Middle Ear P a) Bae ae fee St |] 2
Carbuncle . 4) 2}/—,1 i ss ||
Osteomyelitis ens ee Bee es ee
Total. . , 00°82) 7 3) 45l 6} 3214
Pyamia AND SEPTICEMIA.
When pyogenic micrococci get access to the blood stream they
may be carried into distant parts, and by multiplying produce meta-
static abscesses in the lymphatic glands, bones, joints, and internal
organs, a condition which is recognised as pyemia.
If there is a general invasion of the blood stream by micrococci,
and absorption of their poisonous products, septicemia results, and
death may occur before the development of any secondary lesions.
When septic micro-organisms multiply locally, and their chemical
products are absorbed, or their products are separated from putrid
material and injected into the circulation, the result may be called
sapremia, The blood in septicemia contains living organisms, and is
infective. The blood in sapremia contains only the toxic chemical
products, and is not infective. The one is septic infection and the
other septic intoxication. Pysemia may follow accidental wounds,
surgical operations, parturition, acute suppuration of bones, scarlet
fever, typhoid fever, and other diseases.
To avoid pyemia in surgery and midwifery, the greatest care
must be taken to prevent micro-organisms from being conveyed by
instruments, sponges, bandages, and by the hands of the surgeon
or the obstetric physician. By the use of antiseptics and absolute
cleanliness the chances of infection are reduced to a minimum.
176 INFECTIVE DISEASES.
Rosenbach examined six cases of metastatic pyemia: Strepto-
coccus pyogenes was found five times, partly in the blood and partly
in the metastatic deposits, and twice in company with Staphylococcus
pyogenes aureus.
Baumgarten, also, found Streptococcus pyogenes in the internal
organs in pyeemic cases, and Hiselsberg found Streptococcus pyogenes
in company with Staphylococcus pyogenes aureus in the blood of
cases of septicaemia. '
Frinkel isolated a streptococcus from puerperal fever, which
he at first called Streptococeus puerperalis, but subsequently
identified with Streptococcus pyogenes. These researches have
been confirmed by others. Winkel obtained a pure cultivation of
a streptococcus from the blood of the heart in a case of puerperal
peritonitis. It produced erysipelatous redness when inoculated
in the rabbit’s ear, and in form and in cultivation was similar to
the streptococcus in erysipelas. Cushing also found Streptococcus
pyogenes associated with puerperal infection. The cocci were
, found in endometritis diphtheritica as well as in secondary puerperal
inflammation. These observations were still further confirmed by
Baumgarten, and Bumm isolated the same organism in puerperal
mastitis.
Description oF Bacrerira In Pus.
A description may now be given of the cocci most frequently
found. Staphylococcus pyogenes aureus and albus and Strepto-
‘coceus pyogenes and Gonococcus are the most important of these.
Staphylococcus pyogenes citreus, cereus albus and flavus, are pro-
bably merely epiphytic. Micrococcus tetragenus, Micrococcus
pyogenes tenuis, Bacillus pyogenes feetidus, Bacillus pyocyaneus,
Bacillus coli communis, Bacillus septicus vesice, Urobacillus lique-
faciens septicus, and Bacillus intracellularis meningitidis will be
described fully in Part III. The description of Actinomyces, of
Micrococcus pneumoniz croupose and of the bacilli of anthrax,
tuberculosis, glanders, and typhoid fever, will be found in other
chapters in Part JI.
Staphylococcus pyogenes aureus (Rosenbach).—Yellow
coccus in pus (Ogston). Cocci singly, in pairs, very short chains, and
irregular masses. Cultivated on nutrient agar-agar, an orange-
yellow culture develops, looking like a streak made with oil paint.
One variety grows on nutrient gelatine without liquefying it;
another produces rapid liquefaction, and the growth subsides as
SUPPURATION, PYAMIA, SEPTICEMIA, ERYSIPELAS. 177
an orange-yellow sediment. On potatoes and blood serum a similar
orange-yellow culture grows luxuriantly. They may also form
colourless growths in sub-cultures, and are then indistinguishable
from Staphylococcus pyogenes albus. The cocci do not cause any
septic odour in pus, nor does any gas develop. Albumin is con-
verted by their action into peptones.
They produce rapid ammoniacal fer- :
mentation in urine (Shattock). : ie
The micro-organisms injected into
the pleura or knee of a rabbit produce, = 4
asa rule, a fatal result on the following
day ; but if it surviyes longer, it eventu-
ally dies of severe phlegmon. Tf injected 2 Ges ae ieies
into the knee of a dog, suppuration ip
occurs, followed by disintegration of the ane)
jot. Injected into the peritoneal fy6, 84,—Pus wire STaPHYLo-
cavity of animals, they set up perito- coccr, x 800 (FLUGcE).
nitis, and introduced into the jugular
vein they produce septicemia and death. When a small quantity of
a cultivation is introduced into the jugular vein after previous fracture
or contusion of the bones of the leg, the animal dies in about ten days,
and abscesses ave found in and around the bones, and in sonie cases in
the lungs and kidneys,
and the cocci are found —
in the blood and pus.
Garré caused sup-
puration by inoculating
a pure-culture in ‘a
wound near his finger
nail. Bockhart suffered
from several pustules
after vaccinating his
arm witha pure-culture
suspended in salt solu-
tion, and Bumm gave
himself a hypodermic
Vic. 85.—Suscurannous Tissur ot A Rapsir 48
Hours arrer AN INJECTION OF STAPHYLOCOCCI, x
950 (BAUMGARTEN)
).
injection of a pure-
culture and produced
an abscess. This micro-
organism is practically ubiquitous. It has been cultivated from
the skin and mucous membranes and secretions of healthy persons,
and it occurs in the air, in soil, in dust, and in water, and in
12
178 INFECTIVE DISEASES.
association with suppuration, pyemia, puerperal fever, and acute
osteomyelitis.
Staphylococcus pyogenes albus (Rosenbach).—Cocci micro-
scopically indistinguishable from the above. In cultivations also
they resemble Staphylococcus pyogenes aureus, but the growth
consists of opaque white masses. They, as a rule, liquefy nutrient
gelatine very rapidly, and subside to the bottom as a white sediment ;
more rarely they liquefy very slowly ; and a variety has also been
described which does not produce any liquefaction. They are similar
to the above-mentioned in their pathogenic action. Pure-cultivations
of the organism were obtained from a case of acute suppuration
of the knee-joint.
Staphylococcus pyogenes citreus (Passet).—Cocci singly, in
pairs, very short chains, and irregular masses. If cultivated on
nutrient gelatine or nutrient agar-agar, a sulphur or lemon-yellow
growth develops. When inoculated under the skin of mice, guinea-
pigs, or rabbits, an abscess forms after a few days, from which a
fresh cultivation of the micro-organism can be obtained.
Staphylococeus cereus albus (Passet).—Cocci, morphologic-
ally similar to the above, but distinguished by forming on nutrient
gelatine a white, slightly shining layer, like drops of stearine or wax,
with somewhat thickened, irregular edges. In the depth of gelatine
they form a greyish-white, granular ‘thread. In_plate-cultiva-
tions, on the first day, white points are observed, which spread
themselves out on the surface to spots of 1 to 2mm. When culti-
vated on blood serum a_ greyish-white, slightly shining streak
develops, and on potatoes the cocci form a layer which is similarly
coloured.
Staphylococcus cereus flavus (Passet).—Cocci which produce
in nutrient jelly a growth which, at first white, becomes lemon-
yellow, somewhat darker in colour than Staphylococcus pyogenes
citreus. Microscopically Staphylococcus cereus flavus corresponds
with Staphylococcus cereus albus. Inoculation experiments with
both kinds give negative results.
Streptococcus pyogenes (Rosenbach). << teed occurring singly,
in masses, and in chains. The individual cocci are small spherical
cells, with a special tendency after fission for the resulting elements
to remain attached to each other, forming chains or rosaries. In
cultures on solid media they often occur in the form of staphylococci,
but in liquid cultures there may be a few, three or more elements,
linked together ; or a great number , forming long chains which may
be straight, serpentine, or twisted.
DESCRIPTION OF PLATE IV.
Streptococcus Pyogenes.
Fig. 1.—From a cover-glass preparation of pus from a pyzmic abscess.
Stained with gentian-violet by the method of Gram, and contrast-stained *
with eosin. x 1200. Powell and Lealand’s apochromatic 7, Hom. imm.
E. P. 10.
Fig. 2.—From cover-glass preparations of artificial cultivations of the strepto-
coccus in broth and in milk at different stages of growth. x 1200. Powell
and Lealand’s apochromatic #; Hom. imm. E. P. 10.
In these preparations there is a great diversity in size and form of the
chains and their component elements. In the drawing examples are
figured of the following:
(a) Branched chains.
(®) Simple chains composed of elements much smaller than the
average size.
(e) Chains with spherical and spindle-shaped elements at irregular
intervals. These are conspicuous by their size, and are sometimes
terminal,
(d e) Chains in which the elements are more or less uniform in size.
(f) Complex chains with elements dividing both longitudinally and
transversely, and varying considerably in size in different lengths
of the same chain.
Plate IV
Ce
Pee
pose eet: ’
gone
“eee
Fees!
.
oa”
ae sailded
0
grnmonne sere
i)
Fig 2
STREPTOCOCCUS PYOGENES
EM,Crockshank fecit, Vincent Brooks, Day & Son, lath.
SUPPURATION, PY/EMIA, SEPTICEMIA, ERYSIPELAS. 179
The individual elements composing the chains will be found to
vary considerably in size: here and there in a preparation will
be found a chain composed of excessively small cocci, in another
part the elements will be all on a larger scale, and again in
another part they will be peculiarly conspicuous on account of their
size. So great is the diversity in the size of the cocci in some of the
chains, that one might imagine that there was more than one kind
of streptococcus present in a preparation, until on examining some
of the longest chains it is observed that various sizes are repre-
sented in different lengths of the same chain. Very characteristic
appearances result from the fact that the cocci enlarge and divide
both longitudinally and transversely ; and, indeed, the largest, for
the most part, clearly show a division in two directions, resulting
in the formation of tetrads. In addition to the forms resulting
from the fission of the cocci, there are here and there in a chain,
and sometimes terminally, larger elements, which are spherical,
spindle-shaped, or in the form of a lemon. In the length of the
chains, as in the size of the individual cocci, there is usually great
diversity, In some cases they are composed of only a few, three
or four cocci; in others eight, ten, or twenty. Here and there
an exquisite rosary will extend in a straight line across the field
of the microscope, or be twisted, curved, or serpentine; in some
preparations twisted or entangled strands are observed which are
-composed of several hundred elements. Such chains will be found
to be much thicker in one part than another. Another char-
acteristic appearance is produced by separation of the elements
resulting from fission in the long direction of the chain, by which
lateral twigs or branches are formed. Another character, which
is very striking, may be seen when the individuals in a chain
have become separated; an unstained or faintly stained membrane
may be found bridging across the interval. This will become still
more visible in preparations contrast-stained with eosin.
In plate-cultivations the appearances of the colonies are not very
striking. They appear to the naked eye after three or four days
as extremely minute, greyish-white, translucent dots, which under
the microscope have a slightly yellowish-brown colour. They are
finely granular and well defined. They do uot liquefy the gelatine,
and after several weeks may not exceed the size of a pin’s head.
If the surface of nutrient gelatine solidified obliquely be traced
over once or- twice with a platinum needle bent at the extremity
into a little hook charged with the cocci, a ribbon-shaped film
develops in two or three days. This film is composed of minute,
180 INFECTIVE DISEASES.
greyish-white, translucent dots or droplets, which can be more easily
recognised with the aid of a pocket lens (Fig. 87). According to
the number of organisms sown on the jelly, the dots or colonies
may be completely isolated, or form a more or less continuous
film. The film by reflected light has an iridescent appearance like
mother-of-pearl, but has a bluish or bluish-grey tint by transmitted
light, and with a pocket lens appears distinctly brownish. The
gelatine is not liquefied, and even after several weeks the cultivation
is limited to the inoculated area, and the individual colonies are, as
a rule, not larger than pins’ heads. In gelatine-cultivations of the
same age, but kept in the incubator at 18° C., the colonies get
irregular in form, especially at the margin of the film, and give the
growth an arborescent, fringed, or serrated appearance. Cultivated
on the oblique surface of nutrient agar-agar at 37° C. the growth is
very similar, forming a film composed of minute white colonies
like grains of sand; but the film appears less transparent, is
‘whiter, and the colonies have a greater tendency to get irregular
in form. If inoculated with one tracing of the needle the growth
is scanty, but tends to get thicker in the centre than towards the
margins, which may have a terraced appearance. Inoculated in
the depth of gelatine, there appears after a day or two at 18°C. a
thread-like growth along the track of the inoculating needle. This
delicate growth is found on examination with a pocket lens to consist
of a linear series of extremely minute granules. In a few days’
more, the beads or granules become more marked, but even after
weeks, the cultivation only appears like a string of minute, white,
compact, globular masses or grains. In broth at 37° C. the cocci in
twenty-four hours create a turbidity, and gradually develop beauti-
ful chains varying in length according to the age of the cultivations.
Even in forty-eight hours there may be chains of eight, ten, twenty,
or a hundred elements. After a few days the growth settles down
at the bottom of the tube in the form of a white deposit, while the
supernatant liquid becomes again clear.
Inoculated subcutaneously in the ear of rabbits, they produce in
two days an inflammatory thickening with erysipelatous redness,
or sometimes suppuration.
They may occur in vaccine lymph, as the result of con-
tamination, and Pfeiffer suggested that before calf lymph is
employed for vaccination it should be tested on a rabbit’s ear.
If in two days no rash has been produced, the possibility of the
presence in the lymph of Streptococeus pyogenes or erysipelatis
is excluded.
SUPPURATION, PY/EMIA, SEPTICEMIA, ERYSIPELAS. 181
According to Fliigge and others, after subcutaneous inoculation
of mice with a small quantity of a cultivation, there is no result in
80 per cent. of the animals experimented upon. Sometimes there
is limited pus formation at the seat of inoculation, sometimes the
animals die without any very striking pathological appearances.
They occur in abscesses, pyeemia, and septicemia, and are often
found in diseases such as scarlet fever and typhoid, associated with
septic complications. They have been isolated from air, soil, and
water.
The streptococcus found in erysipelas agrees in description, and
is merely a variety of Streptococcus pyogenes. It has been definitely
established by the researches of Frankel and Freudenberg, and later
by those of the author, Raskin, Prudden, and Bayard Holmes,
that Streptococcus pyogenes is frequently found in scarlet fever and
diphtheria, and in other diseases associated with septic complica-
tions. The author has isolated Streptococcus pyogenes from acute
abscesses, from suppuration after surgical operations, from pyzmia,
from pyemia after scarlet fever, and from purulent peritonitis.
Some of these cultures have been kept up for very long periods,
extending over some years, so that opportunities occurred for a
complete investigation into the life history of this micro-organism.
Variations in the appearances of cultures have been observed when
obtained from the same source. A number of cultures from pus
were prepared on gelatine and agar, made according to the usual
formula, but at different dates, and, therefore, varying slightly in
composition and quality. Sub-cultures were also started in nutrient
gelatine of precisely the same composition, but from primary cul-
tures of the same micro-organism in different media—agar-agar,
milk, and broth. The descriptions of the streptococcus hitherto
published were then found to be inadequate. The different cultures
and sub-cultures presented striking variations in the microscopical and
macroscopical appearances. Some sub-cultures on gelatine, for exam-
ple, exhibited a finely dotted appearance, others showed every variety
in the size, and degree of opacity of the colonies (Fig. 89). Cultures
in broth also, varied in appearance, owing to slight variation in the
composition of the medium, to slight differences of temperature, and
other conditions difficult to determine. The addition of glycerine to
broth materially alters the appearance of the culture. It was con-
clusively proved that minute differences arise from different conditions
of the cultivating media. The author was led to study exhaustively
the streptococcus of acute suppuration in bovines. Primary cultures
of Streptococcus pyogenes from man, and primary cultures from
182 INFECTIVE DISEASES.
a case of purulent peritonitis in a cow, were carried through
sub-cultures under exactly similar conditions.. -Cultivations of the
Streptococcus pyogenes bovis exhibited variations in microscopical
and cultural characters which were even more marked than in the
case of the Streptococcus pyogenes hominis. By selecting certain
cultures from both sources there was a striking similarity if not
identity between them, but, when compared under exactly identical
conditions, there was more difference in cultural characters between
the Streptococcus pyogenes bovis and the Streptococcus pyogenes
hominis than between the Streptococcus pyogenes hominis and the
Streptococcus erysipelatis, and they may therefore be regarded as
distinct varieties (Figs. 89, 90).
Some of the diseases and -conditions in which Streptococcus
pyogenes has been found may be alluded to more in detail.
Spreading Gangrene —From a case of spreading gangrene, which was
identical with Ogston’s erysipelatoid wound gangrene, and regarded by
him as the most intense and dangerous form of erysipelas, Rosenbach
obtained pure-cultivations of a streptococcus by incising the skin of the
limb, and inoculating tubes from the turbid reddish fluid which escaped.
That the streptococcus was identical with Streptococcus pyogenes was
ascertained by comparison with a cultivation derived from pus, of the
mode of growth, and of the effect on animals.
Surgical Fever.—Eiselsberg proved the presence of a streptococcus
in the blood of several cases of surgical fever in Billroth’s clinic. The
organism was identified by cultivation with Streptococcus pyogenes.
Diphtheria.—In three cases of typical diphtheria, Loffler found a
streptococcus. He isolated it by cultivation, found that it was similar
in form, characters on cultivation, and effects after inoculation, to
Fehleisen’s streptococcus of erysipelas. Liffler was not inclined to
regard them as identical, because Fehleisen never found his cocci in the
blood-vessels. Fliigge named the organism Streptococcus articulorum,
and states, that after subcutaneous inoculation or injection of a cultiva-
tion in mice, a large proportion of the animals die, and in the sections
of the spleen and other organs the streptococci are again seen. Baum-
garten investigated the same subject, and decided that the streptococcus
was identical with Streptococcus pyogenes.
Small-pox.—Hlava has established the presence of Streptococcus
pyogenes in the pustules of variola, and Garré found streptococci in the
internal organs in a case of variola hemorrhagica. In a fatal case of
variola complicated with pemphigus, Garré found a streptococcus in
the pemphigus vesicles. Whether it was identical with Streptococcus
erysipelatis Garré left an open question.
Yellow Fever,—Babes observed the presence of streptococci in the
vessels of the kidney and liver in yellow fever. Cultivation experiments
are wanting. It was probably a case of secondary infection with Strepto-
coccus pyogenes.
SUPPURATION, PYAMIA, SEPTICEMIA, ERYSIPELAS. 183
Bilious Fever.—Babes, in a case of fidvre bilieuse typhoide, found
masses of streptococci filling the vessels of the liver, kidney, and spleen,
This was probably another instance of secondary infection with Strepto-
coccus pyogenes.
Measles.—From the blood and inflammatory post-products in measles,
Babis isolated a streptococcus, which he describes as closely resembling
the Streptococcus pyogenes.
Ulcerative Endocarditis —Wyssokowitsch found cocci in the internal
organs in ulcerative endocarditis, and produced the’ disease in animals,
after injury to the valves, by injection of Streptococcus pyogenes and
other organisms. Weichselbaum, by microscopical research and by
cultivation experiments, proved the presence of Streptococeus: pyogenes
in acute verrucous endocarditis. Baumgarten confirmed this. He found
————
ir
Poe eee
pai a
Si
Fic. 86.—ULcErative ENDOocARDITIS : SECTION OF CaRDIAc Muscig, « 700 (Kocu).
Streptococcus pyogenes alone in one case and accompanied by Staphylo-
coccus aureus in another.
Broncho-pneumonia.—Thaon found a streptococcus in the lungs of
children in fatal cases of broncho-pneumonia, complicating measles,
diphtheria, and whooping cough. It was regarded as identical with the
streptococcus isolated by Léffler from diphtheria. Frankel discovered
a streptococcus in the lungs of a case of true croup complicated with
broncho-pneumonia, and by cultivation established its identity witb
Streptococcus pyogenes.
Anthrax.—Charrin found cocci in rabbits, examined some hours after
death from anthrax. These, when isolated, produced death in rabbits
from septicemia, without suppuration. Chains composed of from fifteen
to twenty elements were found in all the organs. This was probably
another instance of Streptococcus pyogenes.
Syphilis.—Kassowitz and Hochsinger found the presence of a strepto-
184 INFECTIVE DISEASES.
coccus in the tissues and internal organs, and especially in the blood-
vessels, in fatal cases of congenital syphilis. These observers regarded
their discovery as having an important bearing on the etiology of syphilis,
but Kolisko pointed out that it was only the result of septic infection
with presence of Streptococcus pyogenes, as had already been established
in scarlet fever.
Cerebro-spinal MeningitisFrom the meningeal exudation of a case
of apparently idiopathic cerebro-meningitis, Banti found Streptococcus
pyogenes and Staphylococcus aureus and albus. The cocci probably
entered through an abscess of the jejunum.
Fic. 87.—Purt-CuLTuRES Or STREPTOCOCCUS PYOGENES.
a, On the surface of nutrient gelatine ; b, in the depth of nutrient gelatine ;
c, on the surface of nutrient agar.
Blepharadenitis and Dacryocystis.—Widmark isolated by cultivations
Streptococcus pyogenes and other organisms from cases of blepharadenitis
and phlegmonous dacryocystis. In phlegmonous dacryocystis Widmark
found Streptococcus pyogenes almost exclusively.
Leukemia.—Fliigge cultivated a streptococcus from necrotic patches
in the spleen of a fatal case of leukeemia. Cultures corresponded very
closely with Streptococcus pyogenes. Inoculation in the ears of rabbits
produced similar results to Streptococcus pyogenes or erysipelatis.
Fliigge calls 1t Streptococcus pyogenes malignus, but concludes that it
is probably dentical with the streptococcus from pus.
SUPPURATION, PYAMIA, SEPTIC/EMIA, ERYSIPELAS. 185
ErysIPELas.
Erysipelas is an acute inflammation of the skin, occurring in
connection with wounds, when it is traumatic, and on surfaces
apparently sound, when it is idiopathic, as in erysipelas of the
face. It is highly contagious in surgical wards, and it gives rise
to rapidly fatal puerperal fever in lying-in hospitals. In such cases
the virus is obviously conveyed from sick to healthy persons by
direct contact, or by instruments and sponges, or by the hand
of the surgeon, physician, or nurse, and possibly by the air.
Fic. 88.—SEcrion oF SKIN IN ERYSIPELAS.
v,v, two lymphatic vessels containing leucocytes and m,m, streptococci ; t, connective
tissue ; a, connective tissue and wandering cells. x 600 (Cornit and Ranvier).
Streptococcus of Erysipelas.—In 1882, Fehleisen isolated a
streptococcus in erysipelas, described the appearances on cultivation,
and maintained that it could be distinguished from the streptococcus
of suppuration. Rosenbach agreed that the two micro-organisms
could be distinguished by parallel experiments, and named the one
Streptococcus pyogenes and the other Streptococcus erysipelatis.
(Fehleisen). Rosenbach asserted that the colonies of the latter
were more opaque and whiter than those of Streptococcus pyogenes
and the growth more marked in the depth of nutrient gelatine,
while microscopically the chains were better marked and larger, and
the individual cocci larger than in Streptococcus pyogenes. Others
who investigated this subject could not distinguisli them with
certainty, either by their morphological or cultural characters or
effects on inoculation. Passet found that inoculation of Strepto-
coccus pyogenes induced a condition very similar to that produced by
inoculation of Streptococcus erysipelatis. Hoffa and Hajek described
minute differences, but Biondi and EHiselsberg failed to confirm these.
186 INFECTIVE DISEASES.
Baumgarten failed to prove any essential difference. Mitchell
Prudden found that Streptococcus pyogenes injected into the sub-
cutaneous tissue of the ears of rabbits, produced in one no effect ;
in four, slight transient redness ; in five, local redness followed by
abscess; in twelve, well-marked erysipelatous redness, followed by
complete resolution in seven, abscess in three, and death in two,
Passet, Biondi, Eiselsberg, Baumgarten, and Mitchell Prudden
concluded that, in their morphological, biological, and pathogenic
characters, so far as animals are concerned, the two organisms are
practically identical.
The author investigated the morphology and cultural characters
of the Streptococcus erysipelatis, which he had isolated from a
typical case. This result cleared up the conflicting statements
which had been made by different observers. By carrying out
absolutely parallel experiments, the, Streptococcus pyogenes and
Streptococcus erysipelatis were unquestionably distinguishable, as
Fehleisen and Rosenbach had asserted. In both cases, however,
inoculation of a trace of a culture from a solid medium produced
only transient redness. Injection hypodermically of a broth-culture
produced in both cases a spreading erysipelatous redness, followed
by suppuration. It was found that primary cultures of the two
micro-organisms, cultivated under precisely the same conditions,
differed in the size and character of their chains, in the size of the
individual elements, in the greater opacity of the colonies of Strepto-
coccus erysipelatis, in a greater tendency to confluence, and in
a more rapid growth. The author found that the difference was
most marked in broth-cultures. Abundant flocculi were formed by
Streptococcus pyogenes ; a powdery deposit with special tendency to
form a granular adhesive film at the bottom of the culture flask,
in the case of the streptococcus of erysipelas. Lastly, they differed
in their power of resisting germicides.
Fehleisen inoculated patients in hospital suffering with malignant
growths, and produced a typical erysipelas with sub-cultures after
an incubation of from sixteen to twenty hours. The disease was
marked by rigors, fever, and general distirbance. Patients who had
recently suffered from erysipelas had an immunity.
Emmerich succeeded in proving the presence of streptococci in
the air of a hospital where erysipelas had broken out. These cocci
in their form, their characters on cultivation, and their inoculation
results, were identified with the Streptococcus erysipelatis. It is not
therefore exclusively parasitic.
Streptococci identical or agreeing very closely in their description
SUPPURATION, PYAMIA, SEPTICAMIA, ERYSIPELAS. 187
with Streptococcus pyogenes, have been found in cattle plague, foot
and mouth disease, strangles, contagious mammitis in cows, and
progressive tissue necrosis in mice, and they will be referred to fully
in subsequent chapters.
EXAMINATION AND CULTIVATION OF STREPTOCOCCI.
Cover-glass preparations can be stained with the watery solutions
of the aniline dyes. In some cases very beautiful preparations can
be obtained by using Neelsen’s solution, and removing excess of
—_
CG. b. c.
Fic. 89.—Srreprococeus Procenes Hominis. Pure-cultures on nutrient
gelatine.
a, Sub-culture from agar. b, Sub-culture from broth.
c, Sub-culture from milk. d, Sub-culture from milk.
stain by rinsing in alcohol. To examine pus, milk, or broth, take
an ordinary platinum needle bent at the extremity into a hooklet.
Dip it into the liquid to be examined, and spread it on a cover-
glass into as thin a film as possible; the preparation is treated
in the ordinary way, that is to say, the film is allowed to dry,
and the cover is taken up with forceps, and passed three times
through the flame with its prepared side uppermost.
Gram’s Method with Eosin.—In this way the streptococci are
stained blue, and stand out in marked contrast to the rest of the
preparation. Use freshly prepared solution. Float the cover-glasses
188 INFECTIVE DISEASES.
on the solution for ten minutes to half an hour, then trarsfer them
to iodine-potassic-iodide solution, until they assume the colour of a
tea leaf; then immerse them in alcohol until they are decolorised ;
dip them in an alcoholic solution of eosin for a few moments, and
then transfer them to clove oil to clarity the film; to remove the
clove oil gently press the cover between two layers of clean filter
paper, then mount in xylol balsam.
A good method for cultivating streptococci is to employ a steril-
ised looped platinum wire, and to spread a droplet, for example, of
pus or blood, over the surface of nutrient agar-agar solidified obliquely.
a,
Cc.
Fic. 90.—Srreprococcus Pyocenes Bovis. Pure-cultures on nutrient
gelatine.
a, Sub-culture from agar, b, Sub-culture from broth.
¢, Sub-culture from milk. d, Sub-culture from milk.
The tubes are then placed in the incubator at 37° C.; the strepto-
cocci will appear in the course of two or three days in the form
of minute dotted colonies. If present alone, and in considerable
quantities, the inoculated surface will exhibit a pure cultivation
consisting of a number of such colonies, whilst a flocculent mass is
observed in the liquid which collects at the bottom of agar-agar
tubes; this flocculent mass will be found to be composed of chains.
From such a tube inoculate a number of the small flasks employed
in Pasteur’s laboratory for cultivations in liquids. In this way
a number of pure-cultivations in milk and broth are established,
which can be readily examined from time to time. From a pure-
SUPPURATION, PYMIA, SEPTICEMIA, ERYSIPELAS. 189
cultivation in broth or agar-agar tubes of nutrient gelatine can
be inoculated. Cover-glass-preparations from the growths on solid
media can be made in the usual way, and stained with either a
watery solution of fuchsine or gentian violet: but to stain prepa-
rations made from milk or broth, or from the liquid in agar-agar
tubes, use the method of Gram; the stain will then be removed,
except from the streptococci, and very beautiful preparations
result.
GoNORRHGA.
Gonorrhea is the result of a catarrhal inflammation of the
mucous membrane of the urethra, vagina, or conjunctiva caused by
a characteristic pyogenic organism discovered by Neisser in 1879.
Gonococcus of Neisser.—Cocci, usually in pairs 1:6 p in
length, ‘8 » in width, and tetrads, with those surfaces of the com-
ponent elements which are in contact, flattened. The elements
are more or less kidney-shaped, and are separated by a clear
unstained interval. They are found free in the pus and also in
the interior of the pus cells. They stain with the aniline dyes,
but are decolorised by Gram’s solution. They do not grow on
the ordinary media, such as gelatine, agar, and potato, in marked
contrast to the common pyogenic cocci; but Bumm succeeded in
obtaining a cultivation by using human blood serum, which was
procured for the purpose from the placenta. They give rise to
a very delicate growth in the form of an almost invisible film,
with a moist appearance, which attains its full development in
a few days. Steinschneider used human blood serum and agar
incubated at 35° C.
Krall recommended either agar with grape-sugar and blood serum,
or the same mixture with the addition of 5 per cent. glycerine.
Others have employed nutrient agar with the surface moistened with
sterilised human blood. More recently Keifer has been successful
with a medium which is prepared in the following way: ascitic
fluid is filtered and sterilised by Tyndall’s process, to this is added
an equal quantity of the following mixture, agar 3-5, peptone 5,
glycerine 2, salt -5 (per cent.). The ascitic agar is solidified in a
Petri’s dish, and the culture incubated at 36° C.
They have also been cultivated in albumin from plovers’ eggs,
and in the fluid obtained from a case of synovitis of the knee joint.
Inoculation of rabbits, dogs, horses, and monkeys, has been
invariably unsuccessful, but sub-cultures produce the disease in
the healthy urethra.
190 INFECTIVE DISEASES.
The cocci are found in pus from the urethra and other mucous
membranes affected by the disease. They have also been found in
urethral and inguinal abscesses in association with Staphylococcus
pyogenes aureus.
Meruop oF STAINING.
(
Cover-glass preparations are made in the usual way, and
double stained with Liffler’s methylene blue, and eosin.
Schiitz recommends floating the cdver-glasses for five or ten
minutes in a saturated solution of methylene blue in 5 per cent.
solution of carbolic acid. They are washed in water, rinsed in very
weak acetic acid, and again washed in water. Safranin may be
used as a contrast stain.
Fic. 91.—Gonococcus x 800 (Bum).
a, free cocci; b, cocci in pus cells ; c, epithelial cell containing cocci.
Eeyprian OPHTHALMIA. a
There are two forms of ophthalmia in Egypt, one associated
with Gonococcus and the other with a bacillus closely resembling
the bacillus of mouse-septicemia, but there are minute differences.
Bacillus of Ophthalmia (Koch and Kartulis). Minute rods
which do not grow on gelatine but readily on blood serum and
nutrient agar, forming a plainly visible, whitish-grey shining growth.
Animals are insusceptible, but cultures produced the disease in the
human conjunctiva in two out of six cases.
CHAPTER XIV.
ANTHRAX.
ANTHRAX is a very fatal malady, and most irregular in its be-
haviour. At one time it attacks only one or two animals, and
at another time it will destroy nearly all the stock on a farm.
Farmers formerly regarded:the disease as non-communicable, and
possibly the result of excessive or improper feeding, or faulty sani-
tation, or of climatic conditions over which no control could be
exercised. It is obvious that so long as the disease was regarded as
the result of unknown conditions, no explanation could be given of
its recurrence from time to time, or of certain animals contracting
the disease and others not, and no measures of any use could be
suggested to cope with an outbreak.
Anthrax has always been more prevalent on the Continent than
in England, and this to some extent accounts for the fact that it has
received greater attention abroad. In France, Germany, Hungary,
Russia, and in India and Persia, anthrax at times produces wide-
spread losses. In Siberia it is still known on this account as the
Siberian Plague.
On the Continent there are certain localities known as anthrax
districts on -account of their reputation for anthrax—for example, in
the Upper Bavarian Alps in Germany and in Auvergne in France.
In 1849, Pollender happened to examine the blood of a cow
after death from anthrax, and discovered peculiar rod-like bodies
among the blood cells. The same observation was made independ-
ently by Brauell and Davaine about the same time, but the greatest
importance must be attached to the publication of Davaine’s further
researches in 1863. Many ridiculed the discovery of bacilli, and
stoutly maintained that they were only blood crystals or accidental
structures of no importance.
For many years very little progress was made, and the statements
of other observers who were able to verify and add to Pollender’s
and Davaine’s discoveries, were still received with scepticism.
191
192 INFECTIVE DISEASES.
Within the last few years a great change of opinion has taken
place. Bacteriologists have investigated the whole subject, so that
at the present day we kuow exactly the cause of anthrax.
Bacillus anthracis (Bactéridie du charbon, Bacillus of splenic
fever, Wool-sorters’ disease, or malignant pustule).—Rods 5 to 20 p
long and 1 to 1:25 » broad, and threads; spore-formation present.
As a thorough knowledge of the life-history of this bacillus is of the
greatest importance, the various steps to be followed in a practical
study of it will be successively treated in detail. Its morphological
Fic. 92.—Bacittus ANTHRACIS, x 1200. Blood corpuscles and bacilli
unstained ; from an inoculated mouse (FRANKEL and PFEIFFER).
and biological characteristics have been very completely worked out,
and it serves as an excellent subject for gaining an acquaintance
-with most of the methods employed in studying micro-organisms.
A mouse inoculated with the bacillus or its spores will die im
from twenty-four to forty-eight hours, or more rarely in from
forty-eight to about sixty hours.
Examination after Death.—The spleen is found to be considerably
enlarged, and may be removed, and examined by making cover-glass
preparations, imoculations in nutrient media, and subsequently
sections.
Cover-glass Preparations —In cover-glass preparations of the
blood of the spleen the bacilli are found in enormous numbers.
Preparations should also be made with blood from the heart and
with the exudation from the lungs and other organs; it will be
DESCRIPTION OF PLATE V.
Bacillus Anthracis.
Fig. 1.—From a cover-glass preparation of blood from the spleen of a guinea-
pig inoculated with blood from a sow. x 1200. Powell and Lealand’s
apochromatic 7; Hom.imm. KE. P. 10.
Fig. 2.--From a section of a kidney of a mouse. Under a low power the
preparation has exactly the appearance of an injected specimen. Under
higher amplification the bacilli are seen to have threaded their-way along
the capillaries between the tubules, and to have:collected in masses in
the glomeruli. Stained with Gram’s method (gentian-violet), and eosin.
x 500.
Fig. 3.—Bacillus anthracis and Micrococcus tetragenus. From a section from
the lungs of a mouse which had been inoculated with anthrax three days
after inoculation with Micrococcus tetragenus, A double or mixed infection
resulted. Anthrax-bacilli occurred in vast numbers, completely filling the
small vessels and capillaries, and in addition there were great numbers
of tetrads. Stained by Gram’s method (gentian-violet), and with eosin.
x 500.
Plate V.
=~ 4
SRY!
We 4
4
‘7/
/,
N
AN Sar
a
‘
a
I)
—
ME
wa
\U
\ Ss,
= (A
~N
Fig On
Fig a.
BACILLUS ANTHRACIS
ANTHRAX. 193
noted that in these the bacilli are present in very small numbers, or
altogether absent. The bacilli should be examined both unstained
and stained. The rods are straight or sometimes curved ; rigid and
motionless, They can be stained with a watery solution of any of
the aniline dyes, and are then seen to be composed of segments with
their extremities truncated at right angles; between the segments
a clear linear space exists, which gives them a characteristic appear-
ance (Plate V., Fig. 1). By double staining, with Gram’s method and
eosin, the rods are seen to consist of a membrane or hyaline sheath
with protoplasmic contents.
Drop-cultures.—A little of the blood from the spleen or heart
may be employed to inoculate sterilised broth or blood serum.
Several of these cultures should be prepared, and some of them
placed in the incubator, and examined at intervals of a few hours.
It will be observed that the rods grow into long homogeneous fila-
ments, which are twisted up in strands, and partly untwisted in long
and graceful curves. ‘The filaments begin to swell,
become faintly granular, and bright, oval spores
develop (Plate 1). The cultures in the incubator
develop rapidly. A temperature of 30° to 37° C.
is the most favourable for spore-formation. The
spores are eventually set free, and by making
a fresh cultivation, or by injecting them into a
mouse or guinea-pig, they germinate again into
the characteristic bacilli, which in their turn
grow into filaments and spores. When the spore
germinates it swells, the envelope becomes jelly-
like, and gives way at one or other pole, and the
contents escape and grow into a rod.
Test-tube Cultivations in Nutrient Gelatine.—
Typically characteristic appearances are’ obtained
by inoculating a 5 to 8 per cent. nutrient gelatine.
A whitish line develops in the track of the inocu-
lating needle, and from it fine filaments spread out
in the surrounding medium (Fig. 93). The fila~ wre. 93,—Purn Cut-
ments are more easily observed with a magnifying TIVATION or Ba-
CILLUS ANTHRACIS
s _ : IN NUTRIENT GE-
appears-only as a thick white thread. As lique- arin,
faction of the gelatine progresses, these appearances
gradually alter, and the growth subsides to the bottom of the
tube as a white flocculent mass. In exhausted culture-media, and
sometimes in the blood, filaments are seen in a state of degeneration.
13
glass. Inamoresolid nutrient gelatine the growth
194 INFECTIVE DISEASES.
This has also been observed in sections of the internal organs of a
rabbit which had been inoculated with the anthrax bacillus and had.
died of septicemia the following morning.
Test-tube Cultivations in Nutrient Agar-agar.—Cultivated upon a
Fic. 94.—Cotonigs or BACILLUS ANTHRACIS, x 80 (FLUGGE).
a, after 24 hours; b, after 48 hours.
sloping surface of nutrient agar-agar a viscous snow-white layer is
developed, but without access of air no cultivation can be obtained,
the bacilli being aerobic. This can be demonstrated by completely
embedding a piece of lung or spleen
pulp containing bacilli, in nutrient
agar-agar (p. 22).
Potato - cultivations. — In about
thirty-six to forty-eight hours a creamy-
white or very faintly yellowish layer
forms over the inoculated surface,
usually with a translucent edge, and
sometimes a strong, penetrating odour
of sour milk.
Plate-cultivations.—From the spleen
Fic. 95,.—IMPRESSION-PREPARA- oy blood of the heart cultivations may
TION OF A COLONY, x 70.
be made in nutrient gelatine on plates.
The colonies develop in about two days, according to the temperature
of the room. They appear to the naked eye as little white spots
ANTHRAX, 195
or specks, which, on examination with a low power of the microscope
and small diaphragm, exhibit two distinct forms. One form, on
careful focussing, has the appearance of a little compact ball of
Fic. 96.—MarGIn or A Cotony, x 250
twisted threads; in the other, liquefaction of the gelatine has
commenced, and the threads spread out like locks or plaits of
hair in the neighbouring gelatine.
characteristic (Figs. 94, 96).
These appearances are perfectly
Cover-glass Impressions.—The plate-cultivations should be also
examined as soon as the colonies
appear, by making cover-glass im-
pressions (Fig. 95). The filaments,
examined with a high power, will be
seen to consist of a number of rods
or segments which are perfectly
regular in form. On the other
hand, filaments from a tube-cultiva-
tion in a solid medium will often be
found to be composed, not only of
rods, but here and there of the so-
called involution-forms (Fig. 97).
From cultures in gelatine and
Fic. 97.—FILAMENTS WITH OVAL AND
IRREGULAR ELEMENTS, x 800.
glycerine agar, very striking preparations are sometimes obtained,
with numerous large spherical and lemon-shaped elements. In! a
196 INFECTIVE DISEASES.
cover-glass preparation from a potato-culture the individual segments
will be found to have a great tendency to be isolated one from the
other, and there is copious spore-formation.
Preservation of Spores.—Spores may be preserved simply by allow-
ing anthrax blood to dry and then sealing it in a tube. The spores
from a potato-cultivation are treated as follows :—The inoculated
surface bearing the creamy cultivation is sliced off in a thin
layer, and is mashed up with distilled water in a glass capsule.
Sterilised silk-thread is cut up into lengths of about a quarter of
an inch, and allowed to soak in the paste for some hours, under a
bell-glass. The threads are then picked out with a pair of forceps,
and laid upon a sterilised glass plate, covered with a bell-glass,
and allowed to dry. From the plate, when’ perfectly dry, they
are transferred to a small test-tube, which can be plugged with
cotton-wool, or sealed in the Bunsen burner.
Examination of the Tissues——The organs should be hardened
in absolute alcohol, and sections prepared and stained by the
ordinary methods. The method of Gram -is the most instructive,
and eosin a very satisfactory contrast stain. The capillaries in
the lungs, liver, kidney, spleen, skin, mucous membrane, etc., will
be found to contain bacilli. In some cases the bacilli are so
numerous that a section under a low power has the appearance
of an injected specimen.
Inoculation of Animals——A. thread containing spores, a drop of
blood from an infected animal, or a minute portion of a cultivation,
introduced under the skin of a mouse or guinea-pig, causes a fatal
result, as a rule, in from twenty-four to forty-eight hours. Sheep
fed upon potatoes which have been the medium for cultivating the
bacillus, die in a few days. Goats, hedgehogs, sparrows, cows, horses,
swine, and dogs are all susceptible. Rats are infected with diffi-
culty. Frogs and fish have been rendered susceptible by raising the
temperature of the water in which they lived. Cats, white rats,
and Algerian sheep have an immunity from the disease.
Attenuation of the Virus.— Toussaint attenuated cultures by
exposing them for ten minutes to 55° C. Pasteur obtained a similar
result by resorting to lower degrees of temperature; and Koch,
Gaffky, and Loffler concluded from their experiments, that from
42° to 43° C. the bacillus was most easily deprived of its poisonous
properties. By cultivating. the bacillus in neutralised broth at
42° to 43° C. for about twenty days, the infecting power is weakened,
and animals inoculated with it (premier vaccin) are protected against
the disease. To obtain a still more perfect immunity, they are
ANTHRAX. 197
inoculated a second time with material (deuxiéme vaccin) which has
been less weakened. The animals are then protected against the
most virulent anthrax, but only for a time. From. a weakened
culture, according to Klein, new cultures of virulent bacilli can be
started, and a culture that can be used as a vaccine for sheep kills a
guinea-pig, and then yields bacilli that are fatal to sheep.
The virulence of the bacillus is also altered by passing the
bacillus through different species of animals. The bacillus of sheep
or cattle is fatal when re-inoculated into sheep or cattle; but if
inoculated in mice, the bacilli then obtained lose their virulence
for sheep or cattle, only a transitory illness results, and the animals
are protected for a time against virulent anthrax.
Exposure to a temperature of 55° C., or treatment with ‘5 to
1 per cent. carbolic acid, deprives the bacilli of their virulence.
Chauveau obtained a similar result by cultivating the bacillus at
38° or 39° C. under a pressure of eight atmospheres. The possibility
of mitigating the virus depends upon the species of animal; rodents
cannot be rendered immune by any known anthrax vaccine. The
nature of the toxic products has been described in a previous chapter
(p. 42).
Meruops oF Srarinine THE BacttLus ANTHRACIS.
Cover-glass preparations of blood, etc., can be stained with a
watery solution of any of the aniline dyes, or with Neelsen’s solution
and subsequent treatment with alcohol (p. 87). The preparations
may be dried and mounted permanently in Canada balsam, but the
typical appearances are best observed in freshly stained specimens
examined in water.
.The sheath and protoplasmic contents can be demonstrated in
cover-glass preparations from the
e ® ~~
blood or spleen which have been . o © v
stained with eosin after the method _ 7 —Z
of Gram.
Spores must be stained by the . ;
special methods already described. A
The most satisfactory preparations
are obtained by double-staining with Fic. 98.—Spores or BAcrLius
Ziehl-Neelsen solution and methy- ANTHRACIS STAINED WITH
. GENTIAN VIOLET, x 1500.
‘lene blue (Fig. 7).
Tissue sections are best stained by the method of Gram, and
after-stuined with eosin, picrocarminate of ammonia, or picro-lithium-
-carmine.
198 INFECTIVE DISEASES.
Origin AND Mope oF SprREAD.
As every outbreak of anthrax is the result of the introduction
into the system of the bacilli, the question naturally arises, how
are they introduced on the farm? Where do they come from? and
what are the channels of infection ?
The spores of the bacilli may get into the soil, and may remain
there in a dormant state for many years. The spores were believed
by Pasteur to be taken up by earth-worms, carried to the surface
and deposited in their castings. Animals grazing are thus liable
to be infected; but Koch’s experiments tended to disprove this
theory. Anthrax has been known to break out among cattle
grazing on a field where several years previously some Russian
hides from infected animals had been buried. By some means or
other the spores may contaminate the grass, and hay imported
from an anthrax district may start the disease on a farm on which
it had never been known to occur. The spores may in a similar
way be introduced with blood manure and bone manure, and with
refuse used as manure. The skin, hair, wool, hoofs, and horns of
infected animals, if soiled with blood, are contaminated by the
bacillus.
Another way in which the disease can be communicated may
be illustrated by the transmission of the disease to man. Those
who handle carcasses, wool or hides of infected animals are liable to
contract the disease. Slight scratches, cuts, bites, and pimples, may
readily be inoculated with the bacilli or their spores. Veterinary
surgeons, butchers, herdsmen, cattle drovers—in fact, all those whose
occupation leads them to cut open or skin cattle, sheep, or horses, or
to handle hides and wool—are liable to fall victims to this disease.
In one case which was brought to the author’s notice, a veteri-
nary surgeon had been called to see a bullock which had died
suddenly in a meadow. A post-mortem examination was made, and
the veterinary surgeon wiped his hands, which were soiled with
blood, on some rough grass, and then washed them in a stream.
The sedgy grass made some small cuts on his fingers, and the
result was that he was simultaneously inoculated with the blood
of the bullock. Local anthrax followed, two of his fingers were
amputated, and he fortunately recovered. In another case a butcher
dressed the carcass of a beast which had died suddenly, and while
doing so scratched a pimple on his neck. An anthrax pustule
developed, and after a very serious illness he also recovered; but
in many cases the attack is fatal. ‘‘ Wool-sorters’ disease” is
ANTHRAX. 199
anthrax of the lungs. Bales of foreign wool contain not only wool
from living sheep, but wool which has been clipped from skins of
dead sheep. If any of the sheep died from anthrax the wool is
sure to be contaminated with blood containing the bacilli, and then
wool-sorters engaged in picking the wool readily inoculate them-
selves through a scratch or pimple, or by inhaling the spores. In
many cases Wool-sorters’ disease is fatal.
A farm may become extensively infected by the living animal.
Blood containing the bacilli may be discharged from the mouth and
nostrils, or be passed with the contents of the intestinal canal
and bladder. The droppings contaminate the pasture or byre, and
spore formation, especially in warm weather, quickly takes place.
From this cause the disease may not only be conveyed to healthy
cattle grazing with infected animals, but fresh cases may occur, year
after year, on the same farm, and if hay is cut and sold off the farm,
other cattle at a distance are similarly infected. If the flooring of
cattle sheds is once soiled by infected animals it is easy to account
for those otherwise mysterious outbreaks which occur when the
cattle are taken in for the winter.
Another source of danger arises when blood from a diseased
animal is washed into brooks or streams, for thus the disease may
be carried to farms in which it was previously unknown.
PREVENTIVE MEASURES.
Early recognition and prompt action are essential to prevent the
spread of any communicable disease.
Unfortunately in the case of anthrax only too often the very
first indication of the existence of the disease is the sudden death
in the pasture or byre of an apparently healthy beast, or possibly
of one or more sheep. Nevertheless, the importance of being able
to recognise any early indications is very great, because an im-
mediate and careful examination should at once be made of the
stock on the farm, and suspicious cases isolated from the rest. The
stock-man may notice that one or two animals tend to keep away
from the others. They look dull and cease feeding, and possibly
shivering may be observed. In horses swelling of the throat may
occur, and in some places there is discharge of blood from the
orifices. Death follows the appearance of these symptoms in a
few hours, and often with startling suddenness, Cattle die rapidly,
but sheep, though rapidly contracting the disease, do not as a rule
die so suddenly.
200 INFECTIVE DISEASES.
The characteristic sign after death is enlargement of the spleen
to three or four times its natural size. It is not only enlarged,
but extremely soft and dark in colour. Blood spots are visible on
the internal organs generally, and the intestine often contains a
quantity of blood. The examination of a drop of blood will show
under the microscope the characteristic bacilli. It is, however,
quite unnecessary to make an elaborate post-mortem examination
in order to satisfy oneself whether the disease is really anthrax or
not. If an animal has died suddenly and has created a suspicion
of anthrax, all that it is necessary to do is to cut cff an ear—or a
foot in the case of a sheep——and make a cover-glass preparation at
the first opportunity. :
A farmer with a case of anthrax must be made to realise the
fact that an enormous quantity of poisonous material has to be
dealt with. In fact, an infected animal is more dangerous when
dead than alive. The owner or person in charge must immediately
notify to a police constable the existence, or even a suspicion of
the existence, of the disease. Prompt measures must be taken
to destroy the carcass and all traces of the blood, and thus to
reduce to a minimum the chance of the disease spreading to the
rest of the stock, and of creating fresh outbreaks in the future.
Every possible precaution must be taken to prevent the blood of
the dead animal from contaminating the pasture, byre, or water
supply. The rest of the stock should be removed from the pasture
or cowshed where the disease has broken out. It is desirable to
give a complete change of food and water, and the whole of the
stock should be examined every day for a week, and any animals
showing a rise of temperature should at once be isolated from the
rest. Preventive inoculation has been recommended to protect
the rest of the stock, but there is not sufficient evidence of the
safety of the process to lead to the adoption of this treatment.
‘Animals ready for the butcher may be removed from the risk of
infection by immediate slaughter. To disinfect the pasture the
best plan is a heavy top-dressing of lime, and after six weeks
stock may be readmitted, though not without some risk. If
year after year cases of anthrax occur on a particular pasture,
the most obvious precaution is to keep stock from it altogether
and convert it into arable land. As roots grown on anthrax-
infected soil have been known to convey the disease, the wisest
course if we have to deal with a small field or comparatively
small tract of land is to throw it out of cultivation or to plant
it with trees. :
ANTHRAX, 201
Disposat oF THE CARCASS.
The surest method to render harmless all the bacilli which
exist in the carcass is burning, but cremation offers practical
difficulties, especially if several carcasses have to be destroyed. In
the case of an animal dying in a town, the local conditions may
render it best to adopt destruction by burning or by means
of chemicals. In such a case the carcass should be covered
with quicklime, and then taken, in charge of an officer of the
Local Authority, to a horse-slaughterer’s or knacker’s-yard, and
destroyed by exposure to a high temperature, or by chemical agents
especially in the vicinity of chemical works. Under the usual
circumstances of death occurring on a farm, fortunately the simple
plan of burtal, with the addition of lime or other chemical agents,
is perfectly efficacious, and even without the use of chemicals éf the
carcass has been left unopened, as the bacilli die rapidly if air is
excluded.
Some experiments carried out by M‘Fadyean clearly indicate the
importance of leaving the carcass unopened.
On July 16th a sheep was infected with anthrax by feeding it with a
virulent culture. Five days later it died, and a microscopic examina-
tion of blood from the ear, immediately after death, showed very many
anthrax bacilli. The carcass was left unskinned and unopened until
July 27th, when the various organs were cut out of the chest and
abdomen and placed in a tin box. The box was then buried at a depth
of about two feet in garden earth, and left there undisturbed until
February 15th, when it was exhumed. The organs had become con-
verted into adipocere, and this was thoroughly mixed up with water and
administered to a sheep. The sheep remained perfectly healthy. In
another experiment a rabbit was inoculated with anthrax on June Ist.
It died on June 3rd, and blood from the ear contained the bacilli. The
rabbit was left unopened for three days, and then placed in a flower pot
and buried in garden earth at a depth of two feet. It was exhumed on
February 15th. The tissues were all destroyed by putrefaction, and the
earth in contact with the bones was administered to a sheep without
conveying the disease or producing any ill effects.
Thus, in the first experiment, the lungs and the intestines, in
‘which spore formation was most likely to occur, were used as a
test, and in the second experiment the entire carcass. In both cases
‘there was destruction or disappearance of the bacilli, and these tests,
therefore, confirm in a very marked way the opinion that prompt
burial of the unopened carcass is a perfectly safe plan to adopt.
202 INFECTIVE DISEASES.
If an animal has died in a meadow, a pit six feet deep should
be dug close to the carcass, and if quicklime can be procured with-
out delay the carcass should be buried with a layer about a foot in
depth beneath it and with about the same quantity to cover it, and
the pit filled up with the excavated soil. ;
If there are any traces of blood where the animal lay, the
contaminated ground should be covered with quicklime or drenched
with strong carbolic acid, and the whole of the site of burial fenced
off for six months. If an animal dies near a brook or stream then
the carcass must be removed for burial to a sufficient distance to
prevent any reasonable probability of contamination of the water.
If death has occurred in the byre, the carcass must be removed
to the nearest and most convenient spot for burial, any fodder or
litter which may have been in contact with the deceased animal must
be destroyed, and the shed and cart and any utensils, hurdles, etc.,
disinfected. ‘For the latter purpose thorough scouring with water
and then washing with limewash is recommended. The limewash
should be prepared immediately before use, and four ounces of
chloride of lime, or half a pint of commercial carbolic acid, be added
to each gallon of limewash.
The following is an illustration of the value of preventive
measures based upon a knowledge of the exact nature of the disease.
A farm on the banks of the Yeo was repeatedly attacked by
anthrax. One morning two sheep died, and other cases followed.
The farmer learnt that his predecessor had buried cattle which had
died of anthrax'on the very spot where the sheep were folded. He
removed his flock, and had no further losses among the sheep, but
he continued to lose cattle grazing in the pastures by the river.
These pastures were occasionally flooded by the Yeo. Another
farmer in the same locality heavily manured a field, and shortly
afterwards anthrax broke out in a most deadly form on his farm.
What was the cause of these mysterious outbreaks? The
explanation was forthcoming, and prevention an easy maiter. The
river Yeo received the washings from the wool factories at Yeovil,
and the pastures were contaminated by anthrax spores in the
deposit which was left behind when the flood subsided. In the
second instance, it was found that the manure used for dressing
the pasture consisted of a quantity of refuse from the wool factories.
Infected wool from foreign countries is one of the principal
sources of the disease in this country, and the remedy is to insist
upon the factories destroying their refuse instead of its being
allowed to contaminate the rivers or to be sold as manure.
ANTHRAX. 203
So long as this source of the disease was unknown anthrax
continued to be spread through the agency of the wool factories.
Anthrax spores may also be introduced with foreign oats, hay,
and manure, so that it is almost impossible absolutely to prevent
the importation of the disease; but the danger of its unlimited
extension and disastrous losses can be minimised, and the com-
munication of the disease to man and to swine entirely avoided by
simple precautions.
ANTHRAX IN SWINE.
The occurrence of anthrax in swine is a subject upon which
there has long been considerable diversity of opinion. Some of the
Fic. 99.—ANTHRAX IN Swine. From a photograph taken during life, showing a
swollen condition of the neck and throat six days after ingestion of part of the
viscera of a bullock which had died from anthrax.
earliest writers on the diseases of animals speak of outbreaks of
anthrax among swine, but whether any or all of these outbreaks
were examples of true anthrax has long been a matter of un-
certainty; for it is well known that diseases quite distinct were
included under the name anthraz.
Menschel states that in an outbreak in which twenty-four
persons were attacked with malignant pustule, many of them from
eating the flesh of beasts suffering from anthrax, pigs which were
fed on the same flesh also became affected, and a woman who ate
some of the diseased pork was subsequently ill.
Roche-Lubin, while apparently accepting the occurrence of
anthrax in swine, taught that the pig resisted inoculation with the
blood of a different species.
204 INFECTIVE DISEASES.
In this country accounts have been published from time to time
of a fatal disease in pigs induced by eating the flesh of animals
which had died of what was described as ‘“‘ blood-poisoning.”
Some very striking cases occurred in the practice of Mr. Wilson,
of Berkhampstead, and were reported in the Veterinarian. A
farmer consulted Mr. Wilson respecting an illness with which his
pigs were affected, stating that two or three were dead and many
others seriously ill. They were strong hogs, ranging from six to
nine months old. On inquiry it was ascertained that the farmer
had lost a beast suddenly about a week previously, that the carcass
had been opened in the yard, and the viscera thrown to the pigs.
Mr. Wilson expressed the belief that the disease was anthrax, and
stated that he found the pigs exhibiting many of the symptoms
observable in cattle, with the additional one of enlargement round
the throat from infiltration of a yellow fluid causing discoloration
of the skin.
Also, in the reports of the Agricultural Department of the Privy
Council thirteen pigs were reported as suffering from anthrax in
1886, and one hundred and fifty-nine in 1887.
But the question arose whether the disease in the pigs was
genuine anthrax or septic poisoning.
Williams says: “The flesh of animals which have died or have
been killed whilst suffering from the disease [anthrax] should not
be used as food either for men, pigs, or dogs, as it is apt to cause
death by blood poisoning”; and Steel writes: “‘ Pigs, dogs, and
poultry should not be allowed to feed on blood, flesh, and ejecta
of anthrax victims,” but no statement is made as to the nature of
the illness produced. No doubt these writers have been greatly
influenced by the opinion of many bacteriologists, for Toussaint
maintained that pigs could not be infected with anthrax, and a
similar view was at one time upheld in this country by Klein,
who stated that pigs were very insusceptible. In Germany also,
pigs have been credited with an immunity from this disease.
In the face of these conflicting statements the author carried
out a series of experiments in order to ascertain the nature of the
disease in swine resulting from the ingestion of the offal of animals
which had died of anthrax; and the result of inoculation with blood
of animals which had died of anthrax, and with pure cultivations
of the Bacillus anthracis.
As a result of these experiments genuine anthrax was produced
in swine (a) by feeding them with anthrax offal; (0) by injection
of blood of a bullock which had died of anthrax; (c) by passing
ANTHRAX. 205
bacilli through the guinea-pig, and transmitting them to swine by
injection of blood from the spleen ; (d) by injecting a pure cultiva-
tion of the anthrax bacillus; (e) and lastly, the anthrax bacillus
was isolated from swine in which the disease was accidentally induced
on a farm, and the disease reproduced by inoculation of guinea-pigs
and mice with blood from the spleen.
The Author's Conclusions.—Swine of all ages can be affected with
anthrax. If the disease is induced by ingestion of anthrax offal,
the tonsils are ulcerated, and constitute the point of access of the
bacilli to the blood. In such cases the characteristic symptom is
Fic. 100.—ANTHRAX IN Swine. From a photograph taken post-mortem. Death
occurred four days after the ingestion of offal from a bullock which had died of
anthrax, and there was well-marked cedema of the throat, cheeks, and eyelids.
enormous swelling around the throat. If the disease is induced by
hypodermic injection, the same cedematous infiltration of the tissues
occurs at the place selected for inoculation. Death may occur
in twenty-four hours, or not until after five or six days. There
is a rapid rise of temperature, usually a rash-like discoloration
of the skin, sometimes loss of power over the limbs, and general
weakness and disinclination to move; the animal may lie helplessly
on its belly, and utter plaintive cries when disturbed. At the post-
mortem the most characteristic feature is the gelatinous cdema
which, in the case of ingestion of offal, is found around the throat.
There is usually congestion of all the organs and engorgement of
206 INFECTIVE DISEASES.
the heart and large vessels, fluid in the cavities of the chest and
abdomen, and enlargement and hemorrhage into the lymphatic
glands. There is in some cases inflammation of the intestines with
submucous and subserous hemorrhages. The spleen may be normal
in size, pale and flabby, and the liver only slightly congested and
friable; in other cases the condition is characteristic, the spleen
is the seat of hemorrhage, causing more or less local enlargement,
which is superficially of a deep purple colour; the liver may also
be greatly congested, very friable, and marked with purple patches.
The examination of the bloodof the heart and spleen for anthrax
bacilli must be carried out with great perseverance and discrimi-
nation, as they are present only in small numbers, and in some
cases have given place entirely to septic organisms. Inoculation
with the blood will produce either typical anthrax, or malignant
cedema or some other form of septicemia. Possibly in the cases
arising from ingestion of offal the ulcerated condition of the throat
affords a nidus and a means of access for septic organisms. It
is also well known that blood in a state of putrefaction may
contain the bacillus of malignant cedema. In the presence of
putrefactive organisms the anthrax bacillus rapidly disappears. If,
therefore, inoculation of guinea-pigs or mice is used as a test for
ascertaining the nature of an outbreak in swine, it must not be
concluded, if Pasteur’s or some other form of septicemia result,
that the disease was not anthrax, while, on the other hand, the
discovery of the anthrax bacillus in the blood of the pig, or the
production of anthrax in guinea-pigs or mice, is positive evidence
as to the nature of the original disease.
Peuch, in France, had obtained similar results by injecting pigs
with anthrax blood and anthrax cultures. He also .carried out
some interesting experiments bearing on public health. The leg of
a pig which had died of anthrax was covered with pounded sea-salt.
Previously to the curing, a slice of the flesh was squeezed in a meat-
press, and the liquid thus obtained was employed for inoculation.
The animals inoculated died of typical anthrax. In six weeks the
curing was considered to be completed, and a slice was cut from the
ham and soaked in filtered water. The juice was extracted in the
meat-press, and employed for the inoculation of four guinea-pigs
and three rabbits. Slight swelling and a certain amount of redness
at the seat of inoculation were the only results. A few drops of
the muscle-juice were added to sterilised broth, and produced a
mixed cultivation of micrococci and motile bacilli. A rabbit and two
guinea-pigs inoculated with the cultivation remained quite healthy.
ANTHRAX. 207
These experiments demonstrated that salting destroys the viru-
lence of the flesh of pigs which have died of anthrax, but in order
to obtain this result the salting must be thoroughly carried out. If
the process be incomplete the flesh is still virulent. Thus the leg
of a pig salted for only fourteen days furnished a juice which
possessed a certain amount of virulence. Out of three inoculated
rabbits, one died in ninety-seven hours of anthrax, and the others
recovered. ‘Three guinea-pigs all succumbed, and a fourth guinea-
pig inoculated with a cultivation from the muscle-juice also died.
Peuch considers that there is danger in consuming flesh which has
not been thoroughly cured.
As it has been clearly shown that pigs may become infected with
anthrax, these animals come under the Anthrax Order of 1886.
This provides for the disposal of the carcass; and although Peuch
has shown that salting destroys the virulence of the flesh of pigs
which have died of anthrax, there can be no doubt that it is quite
right that such animals should be condemned as unfit for food.
Further, the recognition of the occurrence of true anthrax in
swine is an additional reason for condemning the Continental practice
of eating hams, sausages, etc., in the raw state. Indeed, the viru-
lence of anthrax flesh suggests one possible explanation of some
of those obscure cases of meat poisoning which have occurred in
this country. It is possible that the flesh of animals which had
died of anthrax was used in the preparation of sausages, pork-pies,
etc, and that the cooking was not sufficient to deprive the meat
of its poisonous properties.
Equine ANTHRAX.
Veterinary authorities have described “ Anthrax in the Horse,”
but it remains to be seen whether there are not two or more affec-
tions included under this heading. Fleming says: “The most
acute form of anthrax, the apoplectic, is somewhat rare in the
horse, and has perhaps been most frequently observed on the
Continent. Though cases are recorded, but through an error in
diagnosis, under other names in the veterinary literature of this
country, I have only witnessed two cases in England ; though during
the intense summer heat in the north of China I had several.”
The question to which the author is in a position to give a
definite answer is, whether the disease produced by the Bacillus
anthracis ever occurs in the horse. Whether that has been pre-
viously determined, at any rate in this country, it is difficult to say.
208 INFECTIVE DISEASES.
Fleming in describing the pathological anatomy of anthrax in the
horse, says: “The spleen is double and treble its ordinary volume ;
its surface is sometimes bosselated by tumours; its texture is
softened and transformed into a viscid reddish-brown or violet mass,
and the mesenteric glands are infiltrated. The blood in it has
been found to contain bacteridia when examined soon after death.”
Williams, who says that “anthrax in the horse rarely occurs
in this country,” adds, that it is prevalent in India, and is
there termed ‘“Loodiana disease,” and in Africa ‘ Horse-sick-
ness.” But “ Horse-sickness,” from recent researches, is certainly
not anthrax. Williams described a case which occurred in 1879 as
one of anthrax. A carriage-horse died suddenly while in harness ;
“a large black tumour was found in the lungs, and the pulmonary
arteries were engorged with black tarry blood, which, when micro-
scopically examined, was found to contain the bacilli in a most
perfect form, and very numerous indeed.” In 1884, an outbreak
of charbonous fever occurred in Liverpool. Williams proceeded
to investigate the outbreak, and found two horses dead on his
arrival, one having died only a few hours previously. The bacilli
from the blood in this case are figured, and the following statement
made : ‘‘ These bacilli seem to differ from those of splenic fever, being
rather smaller in diameter, and so far as my observations go,
multiply by fission only, not developing spores.”
On the other hand, the author investigated the blood of a mare
which was supposed to have died of anthrax, and on examining
cover-glass preparations of the blood, it was found to contain large
numbers of bacilli with the characteristic microscopical appearances
of anthrax bacilli. To place the question beyond any possible doubt
a number of tubes of agar-agar were inoculated. These, after three
days in the incubator, produced typical cultivations, and on examina-
tion by the ordinary methods and by double-staining, yielded very
beautiful preparations of filaments and spores.
At the same time that the cultivations were prepared, two mice
were inoculated at the root of the tail with a trace of the blood.
Two days afterwards they were both found dead, and with the
characteristic post-mortem appearances, spleen much enlarged, and
anthrax bacilli in enormous numbers.
There can be no doubt that true anthrax occurs in the horse ;
and the author, in 1887, recommended that it should be scheduled
under the Contagious Diseases (Animals) Act, and equine anthrax
has been included in the Anthrax Order of 1895.
More recently Pemberthy has described cases of equine anthrax
ANTHRAX.
209
which he believes to have been the result of infection from feeding
on foreign oats or imported hay.
Preventive Inoculation.—The prevention of anthrax by
means of protective inoculation or vaccination has been attempted
on a very large scale in France, and it is claimed that the results
have been very beneficial to agriculture in that country :—
Animals Mortality.
Total {No.of} Vacci- |. Pirie ———— Total |A verage
No. of | Veteri-) nated Joss | _ loss
Year. Animals | nary after After | After | pnuyin Total. | per | before
Vacci- Re- | Receipt 1st 2nd | Fest oe cent. | Vacci-
nated. | ports. of Vacci- | Vacci- Year nation.
Reports, | nation, | nation, .
1882 | 270,040 112 | 243,199 756 847 | 1,037 2,640 | 1:08 | 10%
1888 | 268,505 103 | 193,119 436 272 784 1,492 | 0°77
1884 | 316,553 109 | 231,693 770 444 | 1,083 2,247 | 0:97
1885 | 342,040 144 | 280,107 884 735 990 2,609 | 0°93
1886 | 313,288 88 | 202,064 652 303 514 1,469 | 0°72
1887 | 293,572} 107 | 187,811} 718 737 968 2,493 | 1-29
Shee
bi 1888 269,574 50 101,834 149 ~ 181 300 630 | 0°62
1889 | 239,974 48 88,483 238 285 501 1,024 | 1°16
1890 | 223,611 69 69,865 331 261 244 836 | 1°20
1891 | 218,629 65 53,640 181 102 17 360 | 0-67
1892 | 259,696 70 63,125 319 183 126 628 | 0-99
18938 | 281,333 30 73,939 234 56 224 514 | 0°69
Total 3,296,815 990 | 1,788,677 | 5,668 | 4,406 | 6,798 | 16,872 | 0-94
: (0°32%) | (0°24%) | (0°38%)
1882 35,654 127 22,916 22 12 48 $2 | 0°35 5%
ia 26,453 130 20,501 17 1 46 64 | 0°31
1884 33,900 139 22,616 20 13 52 85 | 0°37
1885 34,000 192 21,078 32 8 67 107 | 0°50
1886 39,154 185 22,113 18 7 39 64 | 0°29
Oxen
1887 48,484 148 28,083 23 18 68 109 | 0°29
or
1888 34,464 61 10,920 8 4 35 47 | 0°43
Cows
1889 82,251 68 11,610 14 7 31 52 | 0°45
1890 33,965 7 11,057 5 4 14 23 | 0-21
1891 40,736 68 10,476 6 4 4 14 | 0-18
1892 41,609 71 9,757 8 3 15 26 | 0:26
\ 1893 38,154 45 9,840 4 1 13 18 | 0°18
438,824 | 1,255 | 200,962 177 82 432 691 | 0°34
(0-09% )| (0°04% )| (021%)
210 INFECTIVE DISEASES.
The vaccine is supplied, by a company in Paris, in two strengths.
Reports are supplied by veterinary surgeons, and the results have
been tabulated by Chamberland and published, and commented upon
by Cope in a report to the Board of Agriculture (1894), The column
of deaths, in the above table, includes the animals which died from
the vaccination, and those which died from natural infection.
It is claimed that the percentage of losses has been reduced from
10 per cent. to ‘94 per cent. in sheep, and from 5 per cent. to ‘34 per
cent. in cattle. Cope, in the report just referred to, regards these
conclusions as somewhat fallacious, because in order to prove that
the animals inoculated received immunity, it should be shown that
they were subsequently exposed to the risks of natural infection.
This was not the case. But a report obtained from the Bureau in
Paris gives the actual number of animals on each of the infected
farms, and the number which have died of the disease ; and when
compared with Chamberland’s statistics it is evident that nine-tenths
were not on farms where the disease appeared—at least, during
1889-92—and that the deaths from anthrax on those farms where
it was reported to exist were, if anything, higher than they were
supposed to be prior to the introduction of the system of vaccination ;
and in spite of the immense number of animals vaccinated the
official returns obtained from Paris, by Cope, indicate that the
mortality from anthrax, calculated in the ordinary way, remains as
high as ever.
Anthrax in France.
No. of = , :
Year. Owthreas PPE N o pesmi No. Di ae a oe of
1889 618 22,599 1,458 6°5
1890 536 24,073 1,123 47
1891 570 21,356 1,444 6:8
1892 607 28,199 1,581 56
CATTLE,
1889 | — 6,059 700 116
1890 _— 5,365 V1 14:4
1891 — 7,299 849 11-6
1892 | _ 5,058 804 159
SHEEP
1889 : — i 16,540 755 46
1390 | -— | 18,708 352 19
1891 — i 14,057 545 4:2
1892 a ! 23,141 T17 34
ANTHRAX. 211
In Germany, veterinary and agricultural authorities agree that
the results have not met with the success which has been claimed
for vaccination in France. Experiments were undertaken for the
German Government, and in one set of experiments twenty-five sheep
were vaccinated with the first vaccine without an accident, but three
died five days after the second vaccine. In another experiment two
hundred and fifty-one sheep were vaccinated with only one death,
and subsequent inoculation with virulent anthrax proved that they
had immunity.
Six head of cattle were vaccinated without any loss, and six more
were used for a control experiment. Inoculation with virulent virus
proved fatal to the control animals, but the vaccinated were pro-
tected. These, with other animals similarly vaccinated, amounting
in all to two hundred and sixty-six sheep and eighty-three head of
cattle, were then turned out to graze on infected pastures with two
hundred and sixteen unvaccinated sheep as a control experiment.
Within five months four of the vaccinated and eight of the un-
vaccinated sheep died of anthrax, and one of the vaccinated and one
of the unvaccinated cattle.
The result of these experiments led to the following conclusions :—
(1) That the first vaccine is mild and harmless.
(2) That. the second vaccine, even in the hands of experts, is
dangerous and often fatal.
(3) That sheep are more affected than cattle by the injections,
exhibiting fever and other indications of illness.
(4) That cattle and sheep which recover from the vaccination
have an immunity against anthrax when tested by experimental
inoculation.
(5) That vaccinated cattle and sheep tested by exposure to
natural infection by grazing on infected pastures contract the
disease in the ordinary way.
(6) That the time for which immunity is conferred has not been
determined.
In England, Klein tested the vaccine, with the result that animals
either succumbed to the vaccine, or to virulent anthrax after recovery
from the vaccine. Protective inoculation has also been employed in
a few instances by leading agriculturists, but with very unsatis-
factory results.
_ Stamping-out System.—In Germany the conclusion is that
the safest measures are destruction of carcasses and disinfection, and
that inoculation will have no effect in lessening the loss caused by
this disease.
212 INFECTIVE DISEASES,
In England the stamping-out system has been advocated for
many years, and is still regarded as the only reliable means for
suppressing the disease ; and the possible introduction of the disease
among healthy stock by vaccination, and especially in localities in
which anthrax is unknown, would be contrary to the principles upon
which the system is based. These principles are illustrated by the
following extracts from the Anthrax Order of 1895 :—
NoviFICATION.
2.—(1) Every person having or having had in his possession or under
his charge, an animal affected with or suspected of anthrax, shall, with all
practicable speed, give notice of the fact of the animal being so affected or
suspected, to a constable of the police force for the police area wherein
the animal so affected or suspected is or was.
(2) The constable shall forthwith give information of the receipt by
him of the notice to an Inspector of the Local Authority, who shall forth-
with report the same to the Local Authority.
(3) The Inspector of the Local Authority shall forthwith give
information of the receipt by him of the notice to the Medical Officer
of Health of the Sanitary District in which the affected or suspected
animal is or was.
Duty of Inspector to act immediately.
3. An Inspector of a Local Authority on receiving in any manner
whatsoever information of the supposed existence of anthrax, or having
reasonable ground to suspect the existence of anthrax, shall proceed with
all practicable speed 1o the place where such disease, according to the
information received by him, exists, or is suspected to exist, and shall
there and elsewhere put in force and discharge the powers and duties
conferred and imposed on him as Inspector, by or under the Act of 1894
and this Order.
Public Warning as to Existence of Disease.
4.—(1) The Local Authority may, if they think fit, give public
warning by placards, advertisement, or otherwise, of the existence of
anthrax in any shed, stable, building, field, or other place, with or without
any particular description thereof, as they think fit, and may continue to
do so during the existence of the disease, and, in case of a shed, stable,
building, or other like place, until the same has been cleansed and dis-
infected in accordance with this Order.
(2) It shall not be lawful for any person (without authority or
excuse) to remove or deface any such placard.
Wilk of Diseased or Suspected Cow not to be Removed,
5. Where anthrax exists or has existed in any shed, stable, building,
or other place, it shall not be lawful to remove from such shed, stable,
ANTHRAX. 213
building, or other place the milk of any cow which is affected with or
suspected of anthrax.
Removal of Dung or other Things.
6. It shall not bei lawful for any person to send or carry, or cause to
be sent or carried, on a railway, canal, river, or inland navigation, or in
a coasting vessel, or on a highway or thoroughfare, any dung, fodder, or
litter that has been in any place in contact with or used about a diseased
or suspected animal, except with a Licence of the Local Authority for
the District in which such place is situate, on a certificate of an Inspector
of the Local Authority certifying that the thing moved has been, so far
as practicable, disinfected.
Disposal of Carcasses,
7.—-(1) The carcass of an animal which at the time of its death was
affected with or suspected of anthrax shall be disposed of by the Local
Authority as follows :—
(i.) Either the Local Authority shall cause the carcass to be buried
as soon as possible in its skin in some convenient or suitable place
removed from any dwelling house and at such a distance from any
well or watercourse as will preclude any risk of the contamination
of the water therein, and at a depth of not less than six feet
below the surface of the earth, having a layer of lime not less
than one foot deep beneath, and a similar layer of lime above,
the carcass ;
(ii.) Or the Local Authority may, if authorised by Licence of the
Board, cause the carcass to be destroyed, under the inspection of
the Local Authority, in the mode following: The carcass shall
be disinfected, and shall then be taken, in charge of aun officer of
the Local Authority, to a horse-slaughterer’s or knacker’s-yard
approved for the purpose by the Board, or other place so approved,
and shall be there destroyed by exposure to a high temperature,
or by chemical agents.
(2) With the view to the execution of the foregoing provisions of this
Article the Local Authority may make such Regulations as they think fit
for prohibiting or regulating the removal of carcasses, or for securing the
burial or destruction of the same.
(3) Before a carcass is removed for burial or destruction under this
Article it shall be covered with quicklime. In no case shall the skin of
the carcass be cut, nor shall anything be done to cause the effusion of
blood.
(4) A Local Authority may cause or allow a carcass to be taken into
the District of another Local Authority to be buried or destroyed, with
the previous consent of that Local Authority, but not otherwise.
Digging Up.
8. It shall not be lawful for any person, except with the Licence of
the Board or permission in writing of an Inspector of the Board, to dig
214 INFECTIVE DISEASES.
up.-or cause to be dug up, the carcass of any animal that has been
buried.
Disinfection in Case of Anthrax.
9.—(1) The Local Authority shall at their own expense cause to be
cleansed and disinfected in the mode provided by this Article—
(a) All‘ those parts of any shed, stable, building, or other place in
which a diseased or suspected animal has been kept or has died or
been slaughtered ;
(b) Every utensil, pen, hurdle, or other thing used for or about any
diseased or suspected animal ;
(c) Every van, cart, or other vehicle used for carrying any diseased
or suspected animal on land otherwise than on a railway.
(2) The mode of the cleansing and disinfection of such shed, stable,
building, or other place, or the part thereof, shall be as follows :—
(i.) All those parts aforesaid of the shed, stable, building, or other
place shall be swept out, and all litter, dung, or other thing that
has been in contact with, or used about, any diseased or suspected
animal shall be effectually removed therefrom ; then
(ii.) The floor and all other parts of the shed, stable, building, or other
place with which the diseased or suspected animal or its droppings
or any discharge from the mouth or nostrils of the animal has come
in contact, shall be, so far as practicable, thoroughly washed or
scrubbed or scoured with water ; then
(iii.) The same parts of the shed, stable, building, or other place shall
be washed over with limewash made of freshly burnt lime and
water, and containing in each gallon of limewash four ounces of
chloride of lime or half a pint of commercial carbolic acid, the
limewash being prepared immediately before use ; :
(iv.) Except that where any place as aforesaid is not capable of being
so cleansed and disinfected, it shall be sufficient if such place be
cleansed and disinfected so far as practicable.
(3) The mode of the cleansing and disinfection of such utensil, pen,
hurdle, or other thing, and such van, cart, or other vehicle aforesaid, shall
be as follows :—
(i.) Each utensil, pen, hurdle, or other thing, van, cart, or other
vehicle, shall be thoroughly scraped, and all litter, dung, sawdust,
or other thing shall be effectually removed therefrom ; then
(ii.) It shall be thoroughly washed or scrubbed or scoured with water ;
then
(iii.) It shall be washed over with limewash made of freshly burnt
lime and water, and containing in each gallon of limewash four
ounces of chloride of lime or half a pint of commercial carbolic
acid, the limewash being prepared immediately before use. :
(4) All litter, dung, or other thing that has been removed from any
such shed, stable, building, place, van, cart, or vehicle as aforesaid, shall
be forthwith burnt or otherwise destroyed or disinfected to the satisfac-
tion of an Inspector of the Local Authority.
ANTHRAX. 215
(5) The Local Authority may make such Regulations as they think
fit for the purpose of carrying out the provisions of this Article.
Occupiers to give Facilities for Cleansing.
10.—(1) Where the power of causing any place, thing, or vehic'e to
be cleansed and disinfected under this Order is exercised by a Local
Authority, the owner and occupier and person in charge of the place,
thing, or vehicle shall give all reasonable facilities for that purpose.
(2) Any person failing to comply with the provisions of this Article
shall be deemed guilty of an offence against the Act of 1894,
Regulations of Local Authority as to Movement of Animadls,
Fodder, etc,
11. A Local Authority may make such Regulations as they think fit
for the following purposes, or any of them :—
(a) For prohibiting or regulating the movement of any diseased or
suspected animal into or out of any shed, stable, building, field, or
other place, or any part thereof ;
(b) For prohibiting or regulating the movement of any animal into
or out of any shed, stable, building, field, or other place, or any
part thereof, in which there is or has been any diseased or
suspected animal ; and
‘(c) For regulating the removal out of any shed, stable, building,
field, or other place of any fodder, litter, or other thing that has
been in contact with or used for or about any diseased or suspected
animal ; :
but nothing in any such Regulation shall authorise movement in
contravention of any provision of any Order of the Board for the time
being in force ; and a Regulation under paragraph (0) of this Article
shall operate so long only as any animal which in the judgment of the
Local Authority is diseased or suspected remains in the shed, stable,
building, field, or other place to which the Regulation refers, and, in case
of a shed, stable, building, or other like place, until the same has been
cleansed and disinfected in accordance with this Order.
Slaughter in Anthrax and Compensation.
12.-(1) A Local Authority may if they think fit cause to be
slaughtered—
(a) Any animal affected with anthrax or suspected of being so
affected ; and
(6) Any animal being or having been in the same field, shed, or other
place, or in the same herd or flock or otherwise in contact with
animals affected with anthrax, or being or having been in the
opinion of the Local Authority in any way exposed to the infection
of anthrax.
(2) The slaughter of animals under this Articlé shall be conducted
in such mode as will so far as possible prevent effusion of blood.
216 INFECTIVE DISEASES.
(3) The Local Authority shall out of the local rate pay compensation
as follows for animals slaughtered under this Article :-—
(a) Where the animal slaughtered was affected eal anthrax the
compensation shall be one-half of the value of the animal
_immediately before it became so affected ; and
_() In every other case the compensation shall be the value of the
animal immediately before it was slaughtered. ;
“(4) Provided, that if the owner of thé animal gives notice in writing
to the Local Authority, or their Inspector or other officer, that he objects
tothe animal being slaughtered, it shall not be lawful for the Local
Authority to cause that animal to be slaughtered except with the further
special authority of the Board first obtained.
Keeping of Swine in Slaughter-houses.
16. It shall. not be lawful'for any person, in any case in which the
slaughter of any animal is authorised or required by this Order, to use
for such slaughter any slaughter-house in which swine are kept.
Whether an anthrax virus can be obtained which is absolutely
incapable of creating centres of infection, and can therefore be
recommended with safety. for vaccination as an auxiliary and
voluntary measure, is a matter for further investigation.
CHAPTER XV.
QUARTER-EVIL.~-MALIGNANT EDEMA, —RAG-PICKERS’ SEPTICEMIA.-—
SEPTICZMIA OF ODIs BIG: —SEPTICEMIA OF MICE.
QUARTER-EVIL in cattle, malignant cedema, and rag-pickers’ septicemia
in man, septicemia in guinea-pigs, and septicemia in mice, are all
varieties of septicemia produced by bacilli.
An account of quarter-evil, malignant edema, and rag-pickers’
septicemia may appropriately follow the chapter on anthrax, as
they have certain similarities to that disease. They are, however,
not only distinct from anthrax, but must be carefully distinguished
from each other. In connection with these forms of bacillary
septicemia in man and cattle we may study bacillary septicemia
in small animals.
QUARTER-EVIL.
The disease known in this country as quarter-evil or black-leg
is identical with the French Charbon symptomatique and the
German Rauschbrand. Symptomatic anthrax in a very slight degree
resembles anthrax. The disease occurs usually in young cattle from
a few weeks to about twelve months old, and attacks sheep and
horses, but not swine or poultry. It is characterised by the develop-
ment of an emphysematous swelling of the subcutaneous tissue and
muscles, generally over the hind quarter. Infected animals cease
feeding, the temperature rises, lameness supervenes, and death
occurs in about forty-eight hours. The tumour on incision is found
to contain a quantity of dark sanguineous fluid, with characteristic
bacilli.
Bacillus of Quarter-evil (Bucille du charbon symptomatique,
Rauschbrand bacillus).—Motile rods with rounded ends, 3 to 5 w in
length, +5 to 6 » in breadth. Spore-formation present. The
spores are oval, generally situated near the extremity of the rods,
and when fully developed considerably exceed the rods in diameter.
217
218 INFECTIVE DISEASES.
Tnvolution forms are freely developed in old cultures, and in
cultures made in unsuitable media. The bacilli possess numerous
Fic. 101. Bacturr or QuARTER-EVIL x 1000. From
an agar culture (FRANKEL and PFEIFFER).
liquefaction commences. In the depth of
nutrient gelatine the growth occurs in two
or three days at 20° to 25° C. towards the
lower part of the track of the inoculating
needle. ‘The gelatine slowly liquefies, and
there is considerable formation of gas with
the development of a peculiar odour. Spore-
formation occurs freely in cultures, but not
in the blood of infected animals until after
death.
Guinea-pigs inoculated with a pure-
culture, or with spore-bearing threads, die
in twenty-four to thirty-six hours. An em-
physematous infiltration with sanguineous
serum is produced at the seat of inoculation,
and the surrounding muscles are of a dark
colour. The internal organs are more or less
congested. The bacilli are found in the
local exudation and in the surrounding
tissue, and some hours after death in
flagella, and
their
power of movement at
once distinguishesthem
from anthrax bacilli.
They can be cultivated
in the ordinary media
in the absence of oxy-
gen, but more readily
with the addition of
grape-sugar or glyce-
rine. Radiating
ments grow out
fila-
from
the more or less spher-
ical colonies directly
Fie. 102. Pure-CuLTURE OF
BACILLI OF QUARTER-EVIL
IN GRAPE-SUGAR GELA-
TINE (FRANKEL
PFEIFFER):
and
QUARTER-EVIL. 219
increasing numbers in the blood of the heart and in the internal
organs.
Quarter-evil and malignant edema, though possessing points of
resemblance, are distinct diseases. Not only do the bacilli in the
two cases differ in minute morphological and biological details, but
Kitasato showed that guinea-pigs rendered immune against virulent
quarter-evil had no immunity against malignant cedema.
Protective Inoculation.—Arloing, Cornevin and Thomas have
producel iramunity by inoculating healthy cattle with a small
quantity of the fluid from the tumour of an infected animal.
Recovery takes place, and subsequent inoculation with a strong dose
is without effect. Similar results may be obtained by intravenous
injection of a few drops of the exudation, For general application
of the system of protective inoculation, the virulent liquid and
affected muscles are dried at 32° to 35°C., and the dried mass
triturated with water and heated to 100°C. This is used as the
first vaccine.
An infusion similarly prepared, but only heated to 80° C., forms
the second vaccine. The dry powder is a convenient form for
general distribution, and 7, of a gramme is triturated with 5 cc.
of water, and 3 cc. is injected into each animal. In about ten days
the second vaccine is employed, and cattle so treated are said to
have a complete immunity from fatal doses.
The place chosen for the injection is the under surface of the
tail, a short distance from the extremity. The hair is clipped at
this spot, and the point of a syringe is pushed in between the skin
and the bone, and the vaccine slowly injected.
Roux and Chamberland produced immunity by inoculation of
filtered cultures. Cultures in broth were deprived of bacilli by
heating to 115° C., or by filtration through porcelain. Guinea-pigs
were inoculated with three doses of 30 cc. at intervals of two days,
and subsequently injected with a solution of virulent black-leg
powder and lactic acid, which killed control animals in twenty-four
hours. Kitt employed cultures on agar a fortnight old, or fresh
cultures sterilised by steam for thirty minutes. lt was found
possible to confer immunity in oxen, sheep, and guinea-pigs against
the most virulent extract. Kitt’s method has the advantage over
others of only necessitating one single injection. Whether these
experiments are of scientific interest rather than of practical value
may be regarded as an open question.
On the Continent, and especially in France, vaccination against
quarter-evil has been carried out extensively ; and by comparing the
220 INFECTIVE DISEASES.
mortality among the vaccinated and unvaccinated in localities where
the disease commonly occurs, it has been said that the results are
extremely favourable. The matter was investigated in this country
by a committee | of, the Midland Veterinary Medical Association,
and in the course of the experiments some surprising results were
obtained. Six calves and four sheep were vaccinated, and five
calves and two sheep were left unvaccinated as a control experiment.
The seventeen animals were subsequently inoculated with virulent
virus in the form of dried and powdered muscle. In forty-eight
hours all the sheep died, and all the calves exhibited a swelling at
the seat of inoculation. In another set of experiments, healthy
calves inoculated with fresh juice from the tumour in a case of
quarter-evil were not materially affected. The possibility of those
_ealves which possess a natural immunity being classed as protected
by the inoculation must be admitted, and the efficacy and safety
of the process is by no means established.
Matienanr CEDEMA.
The disease known by surgecns as progressive gangrene, gan-
grenous emphysema, or surgical gangrene, has been shown by the
researches of Chauveau, Arloing, Rosenbach and Babés, to be
due to a bacillus identical with the microbe septique of Pasteur and
the bacillus of malignant cedema of Koch. The bacillus or its spores
may be spread by the neglect of antiseptics. The disease occurs
especially after compound fractures and gun-shot wounds.
If a guinea-pig is subcutaneously inoculated with earth, putrid:
fluid, or hay dust, death frequently occurs in from twenty-four to
forty-eight hours. At the autopsy the most characteristic symptom
is a widespread subcutaneous oedema accompanied by air-bubbles.
This originates from the point of inoculation, and contains a
clear reddish liquid full of motile and non-motile bacilli. The
internal organs are little changed, the spleen is enlarged and of a
dark colour, and the lungs are hyperemic, and have hemorrhagic
spots. Examined immediately after death, few or no bacilli are
detected in the blood of the heart, but in that of the spleen, liver,
lungs, and other organs, in the peritoneal exudation, and in and
upon the serous coating of the abdominal organs, they are present in
large numbers. If, on the other hand, the animal is not examined
until some time after death, the bacilli are found in the blood of
the heart, and distributed all over the body.
Bacillus Gidematis Maligni, Koch (Pasteur’s Septicemia).—
MALIGNANT CEDEMA, 221
Rods from 3 to 3°5 pw long and 1 to 1:1 » wide; they mostly lie
in pairs, and then appear to be double this length. ‘The rods are
rounded at their ends, and form threads which are sometimes straight,
but more commonly curved. In stained preparations they have a
somewhat granular appearance. They are motile, possessing flagella,
and form spores. The bacilli are distinguished from anthrax bacilli
by their being somewhat thinner, by their rounded ends, and by their
motility. Moreover, anthrax bacilli never appear as threads in fresh
blood, and are differently distributed throughout the body, They
are anaerobic, and can be cultivated on blood serum and on neutral
solution of Liebig’s meat extract in an atmosphere of carbonic acid.
By embedding material containing bacilli in nutrient agar-agar
7 \ NS |
Fie. 103. Bacirir or Maticnant Gipema x 950. From the subcutaneous tissue
of a guinea-pig. (BAUMGARTEN. )
and nutrient gelatine, characteristic cultivations are obtained. The
following process may be adopted to obtain a pure cultivation. <A
mouse inoculated subcutaneously with dust, as a rule, dies in one
to two days. It is then pinned out, back uppermost, on a slab of
wood, and the hair singed with a Paquelin’s cautery from one hind
leg up to the neck, across the latter, and down again to the opposite
hind leg. Following the cauterised line, the skin is cut through with
sterilised scissors, and the flap turned back and pinned out of the
way. With curved scissors little pieces of the subcutaneous
edematous tissue, in the neighbourhood of the inoculated spot, are
cut out, and sunk with a platinum needle in a 1 per cent. nutrient
agar-agar, or 5 per cent. nutrient gelatine. Fragments of tissue may
also be embedded by the method already described for anaerobic
bacteria.
222 INFECTIVE DISEASES.
The inoculated tubes are placed in the incubator. In a few hours
a whitish turbidity spreads out from the piece of tissue, and upwards
in the needle track. Examined microscopically, the turbidity is found
to be due solely to the development of the bacilli of edema. The
surface exposed to the air exhibits no trace of the bacilli. To
investigate the tubes microscopically, a sterilised glass tube with a
capillary end may be used, with its neck
plugged with sterilised cotton-wool, and
provided at the mouth with a suction ball.
The capillary end is thrust into the cultiva-
tion, and a small fragment removed by
aspiration.
In the course of the first day the bacilli
spread throughout a great part of the
agar-agar in such a way that a more or less
equally diffused cloudiness of the medium en-
sues, with subsequent appearance of strongly
marked clouds or lines of turbidity. At the
same time gas-bubbles develop along the
needle track, and a collection of liquid takes
place, while spore-formation also commences.
The following day these appearances are more
marked, the opacity is more pronounced, the
development of gas increases, and the liquid
contains more spore-forming bacilli and nu-
merous free-spores.
The nutrient-gelatine cultures during the
first day show no macroscopic change, but
Fic. 104. Pure-cunrurn after a few days the piece of tissue is sur-
ov Bacttius or Mauic- rounded with a white halo. This gradually
ee ie ee spreads in all directions, and is apparently
KEL and Prerrer), beset with hairs. The gelatine liquefies, and
the fragment of tissue, degenerated bacilli,
and spores, sink to the bottom. The cultivation is also very
characteristic in a 4 per cent. nutrient agar-agar. If placed in
the incubator, in a few hours a cloudiness forms around the piece of
embedded tissue, which is caused by bacilli gradually spreading in all
directions in the nutrient medium. Mice inoculated from these
cultivations die more quickly than from the original infection from
dust. On potatoes they are cultivated by introducing a piece of liver
or other tissue containing the bacilli, into the interior of a sterilised
potato, and incubated at 38° C. The bacillus is not deprived of its
virulence by cultivation.
MALIGNANT CGEDEMA. 223
The spores of the cedema-bacilli appear to be very widely dis-
tributed. They are found in the upper cultivated layers of the soil,
in hay dust, in decomposing liquids, and especially in the bodies of
suffocated animals, which are left to decompose ata high temperature.
From any of these sources animals can be successfully inoculated.
The bacillus is not only pathogenic in guinea-pigs, rabbits, and
mice, but also in man and in farm animals, including calves but not
eattle. Pure-cultures inoculated in animals produce edema at the
seat of inoculation
without appreciable
gas formation and
without any putrefac-
tive odour. The odour
and frothy effusion
resulting from the in-
oculation of earth are
due to other bacteria,
which are introduced
simultaneously with
the bacilli of malig-
nant cdema. The
spleen is sometimes
slightly enlarged. By Bc)
touching with a cover- " ss
glass the capsule of we. 105. Bacrixr or Matienanr CEpema x 1000.
the spleen, or by ex- From an agar culture (FRANKEL and P¥EIrrER).
amining the serous
effusion, the bacilli are found in abundance ; but if a preparation
is made from the interior of the spleen or from the blood of
the heart, no bacilli will be found until several hours after
death. In this respect there is a marked difference from
anthrax. Another difference is shown in spore-formation, which:
occurs in the living body in malignant cdema, but never in
anthrax. Animals which recover from the disease are said to
be protected.
Protective Inoculation.—Roux and Chamberland produced
immunity by injecting the chemical products in the filtrate obtained
from cultures in broth. The serum from fatal cases will, it is said,
confer immunity on other animals.
There is a variety of this bacillus in soil according to Fliigge,
agreeing in morphological and cultural but not in pathogenic
characters.
224 INFECTIVE DISEASES.
Rag-Pickers’ SEPTICEMIA,
Rag-pickers’ disease has a resemblance to anthrax or wool-sorters’
disease. After death the spleen is found to be enlarged, the in-
ternal organs are congested, and there are hemorrhages on the
serous membranes. Bordoni-Uffreduzzi isolated bacilli which are
quite easily distinguished from anthrax bacilli. They were found
in the blood and in sections of the internal organs.
Proteus hominis capsulatus.—Rods with rounded ends, singly,
in pairs, and in filaments, somewhat smaller than anthrax bacilli,
and often irregular in form. Spore-formation not described ; they
have a well-marked capsule. Colonies are circular, appearing at
first granular, and later possessing a filamentous structure. In
the depth of gelatine they grow in the shape of a round-headed
nail, like a culture of Friedlinder’s pneumococcus. On the surface of
gelatine they form a shining white layer. On agar the growth is
somewhat transparent. On potato a moist, glistening film gradually
spreads over the surface. They do not liquefy blood serum, and
the growth is similar to that obtained on agar. They prove fatal
to mice and dogs, but rabbits and guinea-pigs are not very sus-
ceptible.g Dogs die usually on the second day after intravenous
injection, and after death there is congestion of the internal organs
and of the intestinal mucous membrane. (Edema is produced at
the seat of inoculation in mice. There are hemorrhages in the
lymphatic glands, and congestion of the liver and kidneys. Similar
organisms have been described by Kolb and by Babés in purpura
hemorrhagica.
SEPTICEMIA OF GUINEA-PIGs.
Guinea-pigs and mice sometimes die of septicemia, characterised
by congestion of the lungs, liver, and kidneys, inflamed peritoneum,
pleural and pericardial exudation, congested spleen, and congestion
of the mucous and serous coats of the intestine. Klein isolated
a bacillus from the blood and the internal organs in these cases.
Bacillus of Septiczemia in Guinea-pigs.—Rods with rounded
ends, motile, with pleomorphic forms, cocci, short rods and filaments.
Colonies appear as small, circular, white dots, which enlarge and
become irregular in outline. In the depth of gelatine a white
filament develops, and on the surface the growth rapidly spreads
with a crenated outline. Broth becomes turbid, and after the second
day a copious white sediment is deposited. Spore-formation not
observed.
DESCRIPTION OF PLATE VI.
Bacillus Murisepticus.
Fig. 1.—From a section of a kidney of a mouse which had died after inocula-
tion with a pure-cultivation of the bacillus. With moderate amplification,
the white blood-corpuscles have a granular appearance, and irregular
granular masses are scattered between the kidney tubules. Stained by
Gram’s method with eosin. x 200.
Fig. 2.—Part of the same preparation with high amplification. The granular
appearances are found to be due to the presence of great numbers of
extremely minute bacilli. x 1500.
BACILLUS MURISEPTICUS
E,M.Crookshamk fecit. Vincent Brooks,Day & Son, Lith.
SEPTICEMIA OF MICE. 225
According to Wooldridge, the chemical products of this bacillus,
separated by filtration, produce on inoculation immunity against
virulent bacilli.
SEpTic“mM1a oF Mics.
Mice inoculated with a minimum quantity of putrid fluid often
die of septicemia. They rapidly sicken, their
eyes inflame, their eyelids stick together, they
become soporific, and death occurs in forty to
sixty hours. There is slight edema at the seat
of inoculation, and enlargement of the spleen ;
the bacilli are found free and in the interior
of white corpuscles, both in the edematous
tissue and in the blood capillaries.
Bacillus of Septiczemia of Mice (Koch).
—Extremely minute bacilli, *8 to 1 » long, and
‘1 to -2 » broad, and filaments. Jn cultivations
in gelatine they do not appear to make threads,
but the bacilli lie together in masses. Spores
have been observed, The bacilli are probably
non-motile. They are most commonly in the
interior of white blood corpuscles. In these
they increase, and in many cases a white blood
cell is represented only by a mass of bacilli.
A minimal quantity of blood containing the
bacilli produces the disease if inoculated in
house-mice or sparrows. Field-mice have an
immunity. Rabbits and guinea-pigs inoculated
in the ear suffer only from a local erythema, Te hee pate cae
TIVATION OF THE
renders them for a time immune. Rabbits Bacitius or Sepri-
inoculated in the cornea suffer from an intense C©#MIA or MIcE IN
Nutrient GELATINE,
After two days.
which disappears after five or six days, and
inflammation of the eyes. The bacilli form in
plate-cultivations scarcely perceptible cloud-like
specks, and in a test-tube of nutrient gelatine they form a delicately
clouded cultivation along the needle track.
An identical bacillus has been isolated in swine measles.
(CHAPTER XVI.
SEPTICEMIA OF BUFFALOES.—-SEPTIC PLEURO-PNEUMONIA OF
CALVES.—SWINE FEVER.—SEPTICHEMIA OF DEER.—SEPTICAMIA
OF RABBITS. — FOWL CHOLERA. —- FOWL ENTERITIS, — DUCK
CHOLERA.—GROUSE DISEASE.
THERE are several varieties of septicemia occurring naturally in
buffaloes, deer, calves, and birds, and artificially induced by inocula-
tion of rabbits with septic material. They are associated with
bacteria which agree in their morphological and cultural characters,
though in some cases differing in their pathogenic properties. As
the differences between the bacteria cultivated from these different
sources is not greater than the differences which exist between
the morphological, biological, and pathogenic effects of varieties of
the tubercle bacillus, it will be convenient and fully justifiable to
follow Hueppe and Baumgarten, and regard them as varieties of
the bacillus of hemorrhagic septicemia.
Epipemic Diszasz oF BUFFALOES.
Oreste and Armanni investigated an epidemic among herds of
young buffaloes in Italy (Biiffel-seuche). The disease was extremely
acute, death occurring in from twelve to twenty-four hours. It was
probably identical with an epidemic disease described by Bollinger in
deer. Thesymptoms were fever, rapid pulse, discharge of mucus
from the nose and mouth, and a local swelling of the head and face
leading to suffocation. The only marked feature after death was
hemorrhagic inflammation of the small intestine.
The bacilli were identical with those found by Schiitz in swine
fever. Cultures inoculated in young buffaloes produced the disease.
The bacilli were pathogenic to mice, guinea-pigs, rabbits, pigeons,
and fowls, death taking place in from one to three days.
226
EPIDEMIC DISEASE OF DEER AND BOARS. 227
Septic PLEURO-PNEUMONIA IN CALVES.
Septic pleuro-pneumonia is a disease which attacks young calves
within the first two months after their birth. Percussion and
auscultation reveal lung mischief. The disease is very rapid and
fatal, death occurring on the second or third day. In the less
acute cases one or more lobes of the lungs are found after death
in a state of lobular and inter-lobular pneumonia. The inter-lobular
connective tissue is distended with exudation, giving rise to white
or yellowish bands between the inflamed lobules, which produce a
marbled appearance, recalling the condition of the lungs in infectious
pleuro-pneumonia. The internal organs are congested, and there
are very often hemorrhagic spots on the mucous and serous coats
of the small intestine. All the organs contain rods identical with
those of septicemia of rabbits. Rabbits, guinea-pigs, and mice were
infected. A calf was injected in the pleural cavity with a broth-
culture, and died in twenty hours.
SwinE FEVER.
This disease will be described in a separate chapter. Several
bacteria have been isolated by different investigators. In swine
fever in Germany (Schwein-seuche) Loffler and Schiitz isolated a
bacillus which has been identified with the bacillus isolated by
Salmon and Smith from hog-cholera in America, and with the
bacillus of rabbit septicemia and of fowl cholera.
Eripemic Diszasr or DEER AND Boars.
A. very fatal epizootic (Wildseuche) occurred in the royal game
preserves near Munich, destroying one hundred and fifty-three deer
and two hundred and thirty-four boars (Bollinger). The disease
lasted from twelve hours to six days. In the less acute cases pneu-
monia and pericarditis supervened. In cattle there was also severe
hemorrhagic inflammation of the small intestine. In another form
it produced swelling of the head, face, neck, and tongue. The
virus proved fatal to rabbits in six to eight hours, and to sheep and
goats in about thirty hours. A pig inoculated with a few drops of
blood died in twenty-two hours. Kitt also investigated this malady.
The bacteria were found to be identical, in their appearance and
pathogenic properties, with extremely virulent bacteria from swine
fever. Schiitz distinguished them from the bacteria obtained from
swine fever by their pathogenic effect on pigeons, but cultures
obtained from swine fever do not act uniformly in this respect.
228 INFECTIVE DISEASES.
SEPTICEMIA IN RaABBIts.
Koch minutely investigated a disease of rabbits produced
by inoculation with impure river water and with putrid meat
infusion. Bacteria are found in the blood in abundance, and may
be readily cultivated.
The smallest quantity inoculated subcutaneously or in the cornea
: of a rabbit produces a rise of tempera-
Be oe € 5 ture and laboured breathing after ten
a 1160 J to twelve hours, and death in sixteen
> gy oo to twenty hours, The spleen and
lymphatic glands are found to be
Fic. 107.—Bactentum OF RaBBIT enlarged, and the lungs congested,
Seprice#mia ; Bioop or Spar- :
row, x 700 (Koon). but there are no extravasations, and no
peritonitis. Mice and birds are very
susceptible; guinea-pigs and white rats have an immunity.
Davatne’s SEPTICEMIA.
A disease was produced by Davaine by injecting rabbits with
putrid blood. Rabbits, mice, fowls, pigeons, and sparrows are sus-
ceptible, and guinea-pigs and rats are insusceptible to the bacteria
found in this disease. Rabbits inoculated with a trace of blood con-
taining the bacteria, or with a culture, died in from twenty-four to
thirty-six hours. The spleen, liver, lungs, and intestines are highly
congested, and sometimes extravasations and peritonitis are found.
Fown CHOLERA.
Fowl cholera is an epidemic disease of the poultry-yard much
dreaded in France, and well known through the researches of
Perroncito, Toussaint, Pasteur, and Kitt.
2 a
Fic. 109.—Bacterium or Fowt
Fic. 108.—Bacterium or Fown Cuosra, CHoLEra, x 2500. Muscle juice
> si :
x 1200. From blood of inoculated Fowl. of Fowl.
Fowls suffering from the disease usually die in from twenty-four
to forty-eight hours. The disease shows itself by the fowls becoming
FOWL CHOLERA. 229
somnolent, They suffer from weakness of the legs, and their wings
trail. There is frequently diarrhea, with slimy greenish evacuations,
and death usually ensues after a slight convulsive attack. On
making a post-mortem examination the viscera will be found to be
congested, and there is intense inflammation of the mucous membrane
of the intestine, with hemorrhages.
The blood from the heart, and the intestinal contents, contain
the bacilli which were at one time believed to be peculiar to
this disease. Inoculation subcutaneously, or administration with
food, of a small quantity of a broth cultivation will produce death
in twenty-four to thirty-six hours. Pigeons, pheasants, sparrows,
Fic. 110.—Bacrerium oF Fowi CuHoiera. Section from liver of Fowl x 700
(FLUGGE).
rabbits, and mice are susceptible. In guinea-pigs, sheep, and horses,
an abscess develops at the seat of inoculation. Rabbits are readily
infected by sprinkling a broth-cultivation on cabbage leaves or any
suitable food. It was with this microbe that Pasteur proposed to
eradicate the plague of rabbits in Australia.
Fowl cholera has an additional interest, as it was with this
disease that Pasteur first investigated the attenuation of virus.
Broth-cultures which were several months old were found, when
injected, to produce apparently only a local effect. This weakening
of the virus was attributed by Pasteur to exposure to oxygen.
After recovery the fowls were protected against the action of
virulent cultures, while fowls not immunised died the following day.
Kitt, by working with pure-cultures on solid media, showed that the
230 INFECTIVE DISEASES.
weakening was not due to prolonged exposure to oxygen, but that
old contaminated broth-cultures after a time completely lost their
power, owing to the antagonism of the bacteria accidentally present.
Filtered broth-cultures contain the toxic products of the bacillus,
atid produce slight illness and subsequent immunity.
Fow. Enteritis.
Fowl enteritis is an acute infectious disease of fowls, the course
and symptoms of which are regarded by Klein as distinct from fowl |
cholera. The fowls suffer from diarrhea, with liquid greenish
evacuations, but are never somnolent, and death occurs in one or
two days. After death the-mucous membrane of the intestine is
found to be congested, and coated with grey or yellowish mucus;
the liver is congested, spleen enlarged, and lungs normal. There are
a few bacilli in the blood of the heart, very many in the spleen and
liver, and they are in the form of a pure-culture in the mucus of
the intestine. Klein says that the bacilli are a little longer and
thicker than those found in fowl cholera, which they only slightly
resemble, and that the course of the disease, the symptoms and
pathological appearances, definitely distinguish it from fowl cholera,
but that nevertheless it belongs to the same family of bacilli.
Pigeons are said to be insusceptible, rabbits only slightly susceptible.
By feeding and by subcutaneous inoculation the disease can be
communicated to healthy fowls, but there is no sign of illness until
the fourth day. As regards attenuation, the bacilli behave like
those from cases of fowl cholera.
Duck CHouErRa.
Duck cholera is an epidemic disease of ducks which was investi-
gated by Cornil. The symptoms are similar to those of fowl cholera.
They suffer from diarrhoea and weakness, followed by death in two
or three days.
The bacillus cultivated from the blood of ducks is pathogenic in
ducks but not in fowls or pigeons, and large doses are required to
kill rabbits.
Grouse Disrasz.
Grouse disease is an acute infectious disease of red grouse.
According to Klein the chief pathological feature is severe pneu-
monia ; there is also patchy redness of the serous and mucous linings
GROUSE DISEASE.
231
of the intestine, and the liver is congested and dark, but the spleen
is not enlarged. The bacilli are found in the heart, lungs, and liver,
and in the extravasated blood.
Cultures inoculated in mice and
guinea-pigs produce pneumonia
and death.
ceptible, and other small birds.
Fowls, pigeons, and rabbits are
insusceptible.
Bacillus of Hzmor-
rhagic Septicemia.—Very
short rods, with rounded ends,
‘6 to ‘7 w in width and 1:4 » in
Sparrows are sus-
length.
Fic. 111.— Bacittus or Hamorrwacic
Sepric#MiA. Blood of a Rabbit after
death from Septicemia x 950 (BauM-
GARTEN).
In stained preparations the rods
Fie. 112. — Bacitnus or
Heorruacic SEepriceMia
(Rabbit Septiczemia). Pure-
culture in Gelatine after
four days (BAUMGARTEN. )
been obtained by different observers.
are observed to be deeply stained at the
ends and to have a clear interval in the
middle; they were on this account mis-
taken by earlier observers for dumb-bell
micrococci or diplococci. They are non-
motile, and spore-formation is unknown.
They grow readily in the ordinary media.
The colonies in nutrient gelatine appear
about the third day. They are circular
in form, with a sharp dark outline, and
of a yellow colour, lighter at the peri- -
phery. Later, the central zone is finely
granular, and of a dark yellowish-brown
colour, with the lighter peripheral zone
more clearly defined. In the depth of
gelatine a delicate filament develops in
the track of the needle, composed of
minute spherical colonies, somewhat trans-
parent, and yellowish-white in colour.
At the point of puncture there may be no
growth visible, or a flat and very limited
growth. Inoculated on the surface of
nutrient media a thin layer develops,
with an irregular serrated and thickened
On potato different results have
Some maintain that a
border.
greyish-white or yellowish film will develop at the temperature of
the blood ; but aecording to Caneva, the bacilli, whatever their source,
232 INFECTIVE DISEASES.
will not grow on potato, while Bunzl-Federn maintains that the
bacilli from fowl cholera and rabbit septicemia do grow upon potato,
but those from septicemia in deer, buffaloes, and swine do not.
Opinions differ with regard to their action on milk. The reaction
for phenol and indol is given in all cases, except with cultures
obtained from septicemia of buffaloes. The virulence of the bacilli
may be diminished and attenuated, but it may subsequently be
restored by successive inoculation in animals. The pathological
lesions vary in different animals. The most common result is con-
gestion of the internal organs and hemorrhage. The bacilli culti-
vated from cattle or deer produce fatal results when inoculated
in swine. The bacilli from any of these sources inoculated in
pigeons will produce fowl cholera, but the bacilli isolated by Schiitz
from swine, and those from deer, are not fatal to fowls. Further,
the bacilli cultivated from swine fever are fatal to guinea-pigs,
while the bacilli from rabbit septicemia have very little effect. upon
them. The bacilli have been found in association with diseases of
cattle, swine, deer, birds, rabbits, and mice, and have been cultivated
from healthy mucous membrane. Veranus Moore found the bacilli
in the mucus from the upper air passages, of 71 per cent. of cattle,
85 per cent. of cats, and 33 per cent. of dogs. From these sources
inoculations were made in rabbits, and rapidly fatal septicemia
was produced, associated in less acute cases with peritonitis, pleurisy,
and pericarditis.
CHAPTER XVII.
PNEUMONIA.—INFECTIOUS PLEURO-PNEUMONIA OF CATTLE.-—
INFLUENZA.
Acute Croupous PNEUMONIA.
Pyevumonia is an acute inflammation of the lungs with fibrinous
infiltration of the air vesicles and interstitial tissue. There are
varieties of pneumonia, and one form is commonly believed to be
infectious,
The lung passes through three stages—engorgement, red hepati-
sation, and grey hepatisation. In the first stage the lung is of
a deep red colour, but still vesicular; in the second stage the
affected part is*more or less solid, and has the consistency of liver,
owing to the fibrinous lymph which is poured out into the alveolar
cavities. In the grey hepatisation, the exudation contains more
leucocytes and less fibrin, and this is followed by the stage of
suppurative softening and final absorption. The sputum at the
commencement of the disease is rusty, from the presence of blood,
and later on has the appearance of prune juice. Examination of
the sputum by Gram’s method will reveal numerous micro-organisms,
and two of these are deserving of special study—the pneumococcus
of Friedlinder, which is present in a considerable proportion of
cases, and Sternberg’s micrococcus, which was found in sputum by
Talamon. --
1n 1888, there was considerable prevalence of pneumonia in
Middlesbrough, with strong tendency to occur in groups of cases;
but there was admittedly room for doubt whether the clinical
and post-mortem appearances were not identical with ordinary
pneumonia. Dr. Ballard maintained that there were facts and
considerations which appeared to show that the disease was com-
municable from the sick to the healthy, and that it was a specific
febrile disease, and Klein isolated and described the micrococcus
present in these cases.
Bacterium Pneumoniz Crouposze (Pneumococcus, Fried-
233
234 INFECTIVE DISEASES.
lander).—Cocci ellipsoidal and round, singly, or in pairs (diplococci),
rods and thread-forms. The cell-
membrane thickens, and develops
into a gelatinous capsule, which is
hel round if the coccus is single, and
| ellipsoidal if the cocci occur in pairs
or in rod-forms. Cultivated in a
YF GG a test-tube of nutrient gelatine they
! @. | grow in the form of a round-
headed nail, without liquefaction
of the gelatine (Fig. 114). The
cocci when artificially cultivated
Fic. 113.—Bacrertum PNeumonre have no capsule, but it again
Crovuros#, From Preurat Caviry appears after their injection into
or A Mousz, x 1500. A, B. animals,
Thread-forms. C, D, E. Short
rod-forms. G. Diplococci. H.
Cocci. I. Streptococci. (Zopf.) can also be
cultivated
on blood serum and on_ boiled potatoes.
They occur in pneumonic exudation. In-
oculation of dogs with a cultivation of the
cocci occasionally gave positive results; but
in rabbits no results followed. Guinea-
pigs proved to be susceptible in some cases ;
but thirty-two mice, after injection of a
cultivation diffused in sterilised water, into
the lungs, died without exception. The
lungs were red and solid, and contained the
cocei, which were also present in the blood,
and in enormous numbers in the pleural
exudation. Inhalation experiments by spray-
ing the cocci diffused in water into mouse
cages produced pneumonia and pleurisy in
three out of ten mice.
The nail-shaped cultivation is not always
produced, nor are these conclusions accepted
by all investigators.
The cocci
Fic. 114.—FRreDLANDER’S
Pyevumococcus. Pure-
culture in nutrient-
METHODS OF STAINING FRIEDLANDER’S gelatine four days old
(BauMGaRTEN).
PNEUMOCOCCUS.
Cover-glass preparations of pneumonic sputum or exudation may be
treated as follows :—
PNEUMONIA, 235
(a) Stain by the method of Gram, and after-stain with eosin. ; ;
(>) Treat with acetic acid, then stain with gentian-violet or Bismarck-
brown. Examine in distilled water, or dry and preserve in Canada
balsam.
(c) Float them on weak solutions of the aniline dyes twenty-four
hours ; differentiation between coccus and capsule is thus obtained.
(d) Stain with osmic acid; the contour of the capsules is brought
out.
Fic. 115.—Carsvre-cocct rrom Pneumonia, x 1500 (BauMGARTEN).
Sections of pneumonic lung should be stained by—
(a) Method of Gram.
(6) Method of Friedlander. This method is employed to demonstrate
the capsules in tissue sections. It consists in placing the sections twenty-
four hours in the following solution :—
Fuchsine . ‘ R 1
Distilled water i F 100.
Alcohol . ‘ A F A 5
‘ i 04 2
They are then rinsed with alcohol, transferred for a couple of minutes
to a 2 per cent. solution of acetic acid, and treated with alcohol and oil
of cloves in the usual way, and preserved in Canada balsam:
Sternberg’s micrococcus was first found in the blood of rabbits
inoculated with saliva. Three months afterwards, Pasteur encoun-
tered the same organism in rabbits inoculated with the blood of a
child suffering from rabies. The same organism in 1883 was found
by Talamon in pneumonic sputum. It was identified by Sternberg.
Two years afterwards further observations were made by Frinkel,
Gamaleia, and others. It has also been found in purulent meningitis
by Netter, and by Monti in cerebro-spinal meningitis, by Weichsel-
baum in ulcerative endocarditis, and by others in acute abscess of
the middle ear, and in purulent inflammation of the joints following
pneumonia.
Glacial acetic acid .
236 INFECTIVE DISEASES.
Sternberg’s Micrococeus. (Microbe de salive, Pasteur ; Micro-
coccus Pastewri, Sternberg; Lancet-shaped micrococcus, Talamon ;
Streptococcus lanceolatus Pastewri, Gamaleia ; Diplococcus pmneur
monie, Weichselbaum ; Bacillus septicus sputigenus, Fliigge ;
Micrococcus of sputum septicemia, Frankel.) Spherical or oval
cocci, singly, in pairs or in chains, often lanceolate and capsuled.
Stain readily with the aniline colours and by Gram’s method ;
non-motile. They flourish in alkaline media in the incubator. In
broth they produce in twelve hours a cloudiness due to the develop-
Fic. 116.—Micrococctus or Sputum Srepricem1a. From the blood of a
Rabbit. x 1000 (FRANKEL AND PFEIFFER).
ment of cocci and short chains. After a time these subside to
the bottom of the tube, and the liquid above becomes clear. In
plate-cultivations the colonies are small, circular, white, and granular.
In the depth of gelatine, minute white colonies develop along the
track of the needle without liquefaction of the gelatine; and on
the sloping surface of nutrient agar or blood serum minute trans-
parent drops appear along the line of inoculation. They grow in
milk, coagulating casein; but they do not grow on potato. Sub-
cultures quickly lose their virulence, but regain it by inoculation.
oe ad
PNEUMONIA. 237
The injection of a minute quantity (-2 cc.) of a virulent culture
subcutaneously proves fatal to mice and rabbits in from twenty-
four to forty-eight hours. Immediately afterwards there is a rise of
temperature of 2° or 3° C., later it falls, and just before death
it is several degrees below normal. After death, the post-mortem
appearances of septicemia are observed, in addition to diffuse
inflammatory cedema extending in all directions from the point of
injection. The subcutaneous connective tissue contains sanguineous
serum and micrococci in abundance. The liver and spleen are some-
Fic. 117.—Cotontes or STernBere’s Micrococcus. Agar plate-cultivation,
after 24 hours. x 100 (FRANKEL AND PFE£IFFER).
times dark and engorged, and blood from the heart and internal
organs teems with micrococci.
There is no indication of pneumonia after subcutaneous inocula-
tion, but intra-pulmonary injections produce fibrinous pneumonia,
often fatal (Talamon, Gamaleia). The result is usually fatal in
rabbits and sheep, but dogs, as a ru Injection of cultures
into the trachea of rabbits is
(Monti).
Sternberg concludes that t
infectious pneumonia, but the
with widely different pathologica
being a saprophyte, which finds in pneumonia a suitable soil for its
development, must not be overlooked.
238 INFECTIVE DISEASES.
Klein’s Micrococcus.—Klein found in pneumonic sputum a
diplococeus which does not appear to differ from Sternberg’s micro-
coceus. In cover-glass preparations the bacilli are surrounded with
a halo, but no definite capsule, as in Friedlinder’s coccus. They
appear as short rods constricted in the centre, or dumb-bell forms,
and forms intermediate between cocci and bacilli. In gelatine, after
two or three days, greyish-white spots appear, which enlarge in the
next two or three days into flat, translucent, greyish-white plaques,
with irregular serrated outline. Colonies beneath the surface are
spherical, and of a brownish-yellow colour. In test-tubes in the
depth of the gelatine a whitish-brown filament develops on incuba-
tion, composed of minute spherical colonies, and on the surface the
growth spreads out into a greyish-white film with serrated margin.
On the surface of obliquely solidified gelatine the growth forms a
thin whitish film, which enlarges in breadth with irregular outline,
reaching its maximum in about a fortnight. The growth on agar
is very similar. Broth becomes uniformly turbid in twenty-four
hours, then a powdery precipitate makes its appearance. On potato
there is a thin, moist, faintly yellowish-brown film. Cultures examined
in the fresh state show many rods in a resting stage, and others
actively motile. In addition to the dumb-bell forms there are others
of greater length, and in old cultures involuted and degenerated
forms. Spore-formation has not been observed. A broth-eulture
Mis sctuted into two rabbits producer tumour which subsided
in,a week. Death ensued in one case’ in eight days, and in the
other in three week8S. There was purulent matter at the seat of
inoculation in one ; in the other, pericardial exudation and hyperemia
of-the lungs. Broth-cultures inoculated intravenously produced no
effect. In guinea-pigs there was swelling at the seat of inoculation,
or slight~indication of disease and recovery. Cultures inoculated
in mice produced rapid breathing, drowsiness, and death in from
twenty-four to ninety-six hours. The internal organs were_con-
gested, the lungs inflamed, and the blood and organs in the iroetNtel
animals contained the diplococci in consfderable numbers.
isolated a coccus whichghe,named the Micrococcus lanceolatus
Animals either rapid septicemia
Jat a later period.
Inocula Immunity has been produced in
rabbits by the int 101 Pof the virus in a diluted form.
Blood obtained from immunised fabbits was kept at 10° C. for twelve
hours, and then filtered, and animals injected with it acquired
immunity against virulent cultures (Emmerich).
INFECTIOUS PLEURO-PNEUMONIA. 239
Filtered cultures are said to confer immunity for six months, and
raising the temperature of filtered cultures increases the strength of
the substance which gives immunity (Klemperer). The blood serum
of immune animals can confer immunity on other animals, and, it is
said, will arrest the progress of the disease produced by injection
of healthy animals with virulent cultures. The cultures contain a
proteid body, for which the name pneumo-toxin has been suggested,
and anti-pneumo-toxin has been isolated from immunised blood
serum.
Inrectious PLEURO-PNEUMONIA.
Infectious pleuro-pneumonia is a highly infectious disease
peculiar to cattle; it is characterised by rise of temperature and
exudation into the lungs. It is often fatal, and sometimes exists in
an extremely chronic form. It is believed to have been unknown in
England previously to 1840, and is supposed to have been introduced
from Holland, where in one year it destroyed seven thousand cattle.
Fic. 118.—Acutre CatTarrHaL Pneumonia (Ox).
a, Coagulated mucus with catarrhal cells (c) embedded ‘in it; b, catarrhal cells
sprouting from alveolar wall, . x (480. (Hamilton. )
The disease cannot be conveyed “artificially. A living, diseased
animal must be the medium of infection. The disease is apparently
only communicated by cohabitation. Brown injected large quanti-
ties of lymph from diseased lungs into the jugular vein, into the
240 INFECTIVE DISEASES.
tissue of the lungs, and into the trachea, without any result except
a small abscess at the seat of puncture. Administration of the
virus by the mouth gave equally negative results. The lungs from
a recently killed animal infected with pleuro-pneumonia were placed
in a shed occupied by healthy heifers, and left there for several days.
Fodder, litter, and manure were taken from places in which there
were diseased cattle, and placed in contact with healthy cattle, and sub-
sequently all the animals used in these experiments were slaughtered
and carefully examined, and the results were absolutely negative.
Similarly negative results followed experiments made by Sander-
Fig. 119.—Inrectious PLEURO-PNEUMONIA OF CATTLE, x 480.
a,a,a, Exudation in air-vesicles, composed of a network of fibrinous lymph with
entangled leucocytes; b,b, the same caseating; ¢, the air-vesicle filled with
leucocytes only. In the centre is a blood-vessel filled with a fibrinous plug.
(Hamilton. )
son and Duguid,.and thus confirmed the conclusion arrived at by
Brown, that the disease could only be communicated by actual contact
of a living, diseased animal with a healthy one.
The symptoms of the disease in cattle are a rise of temperature
to 105° or 107°, and a peculiar dry cough, and later the usual
indications of pneumonia, difficulty in breathing, and dulness on
percussion. Asarule, death follows from exhaustion ; but the disease
may also assume a chronic form, if the animal escapes slaughter,
and the lung may become gangrenous or tubercular. The period of
incubation is about thirty days, but it is uncertain. The lesions are
ft pees EA
Ce SAR fa
BNW NLA
¢, a ) Ai i
Fic. 120.—Inrecrious PLEURO-PNEUMONIA Or CaTTLE, x 50.
a,a,a, Spaces in deep layer of pleura and interlobular septa filled with fibrinous
lymph; }, deep layer of pleura running down to an interlobular septum ; ¢,¢,
air-vesicles filled with fibrinous lymph; d, blood-vessels of alveolar walls,
much congested; ¢, large congested blood-vessels; ff, interlobular septa
infiltrated with fibrinous lymph;! g, blood-vessel in interlobular septum
(Logwood, Eosin and Farrant’s solution).—Hamitton.
16
242 INFECTIVE DISEASES.
limited almost entirely to the lungs; congestion is quickly followed
by inflammation and effusion into the air vesicles and the intra-
lobular fibrous tissue which is so well marked in the lungs of cattle.
Leucocytes are entangled in the fibrinous lymph, and the intra-
lobular septa are enormously enlarged, so that the red lobules are
mapped out by the paler septa, and produce on section of the diseased
parts a very striking marbled appearance. A somewhat similar
appearance is sometimes observed in septic pleuro-pneumonia in
calves. The effusion occurs also in the air vesicles. The stages
of grey hepatisation and suppurative softening have not, as a rule,
time to develop. Hemorrhagic infarctions are sometimes produced,
which in turn become gangrenous or cheesy, and a capsule may form
round the diseased part. Roy found micro-organisms in the lymph,
but attached no importance to them. Bruylants and Verriet also
described a micro-organism in the lymph. Later, Poels and Nolen
isolated a micrococcus resembling Friedlinder’s pneumococcus,
Inoculation in the lungs produced a condition in cattle which they
considered indicative of pleuro-pneumonia.
Lustig was unable to confirm these observations, but succeeded
in isolating from lymph a bacillus and three species of micrococci.
‘One of the micrococci formed an orange growth when cultivated,
and was regarded as the specific micro-organism, as it caused sub-
cutaneous tumefaction, and, it is said, some degree of immunity.
Brown cultivated a number of organisms which on inoculation
only produced local irritation. Intravenous injection produced death
from septicemia in one case in thirty-six hours.
Arloing isolated four different organisms, including a bacillus
which was named Pneumo-bacillus liquefaciens bovis. Later, he
prepared a fluid from broth-cultures, pneumo-bacillin, which pro-
duced a more marked rise in temperature in animals suffering from
pleuro-pneumonia than in healthy animals, and its use was
suggested as an aid in diagnosis. Arloing named the micro-
organisms provisionally Pneumo-bacillus liquefaciens bovis, Pneumo-
coceus gutta cerei, Pneumococcus lichenoides, and Pneumococcus
flavescens.
Pneumo-bacillus liquefaciens bovis.—Short rods, non-
motile; spore-formation not observed. They rapidly liquefy gelatine,
and form on potato a white layer, which becomes brownish and
sometimes greenish. According to Arloing pure-cultures produce
in the ox, when injected subcutaneously or in the lung, the same
lesions which are produced by virulent lymph. Guinea-pigs and
rabbits are slightly susceptible, dogs are immune.
INFECTIOUS PLEURO-PNEUMONIA. 243
Nocard does not accept Arloing’s conclusions, and expresses the
opinion that the virus is particulate, but is not due to any micro-
organism. which can ,be detected or cultivated by the methods
at present adopted. In the opinion of the author, who has
also examined the micro-organisms in pleuro-pneumonia, it is
fully justifiable to regard the nature of the contagium as
‘unknown.
Preventive Inoculation.—In 1852 Willems introduced inocula-
tion, The liquid from the lungs of an animal with pleuro-pneumonia,
which had recently died, was inoculated in the extremity of the tail
by a puncture with a lancet. Swelling occurred at the seat of inocu-
jation, and on recovery the animals were believed to be protected.
‘A Dutch Commission reported that the inoculation gave a temporary
protection. A Belgian Commission in the following year reported
that the phenomena of inoculation could be produced several times
in succession in the same animal, and that it was not a certain
preventive. A: French Commission in 1854 concluded that a power
of resisting infection was given, but the period was undetermined. -
Protective inoculation continued to be employed, and various modi-
fications of the method were introduced. Threads soaked in lymph
were inoculated, or the lymph subcutaneously or intravenously
injected.
The usual result of the inoculation is swelling and, in about ten
or fourteen days, effusion of straw-coloured fluid, which is occasion-
ally blood-stained. Gangrene may follow, involving amputation of
the tail. Germont and Loire in Queensland adopted the method—
which was suggested by Pasteur—of inoculating calves in the loose
cellular’ tissue behind the shoulder. This produces intense edema
and a quantity of lymph. There has been much controversy with
regard to the value of protective inoculation. ey
Stamping-out System.—Brown maintains that pleuro-
pneumonia can be exterminated only by slaughter of the diseased
animals, and quotes the results experienced in the Netherlands in
support of his views.
In 1871 slaughter for pleuro-pneumonia was commenced in the
Netherlands. There were 6,000 cattle attacked by the disease. In
1872 owners were compelled to slaughter not only diseased cattle,
but those which had been in contact with them, unless inoculated,
and the attacks were, in consequence, reduced to 4,000. In 1873 it
was forbidden to move cattle out of infected districts, and the attacks
were reduced to 2,479. In 1876 slaughter of the whole herd was
decreed, and during the first year of this heroic system the cases fell
244 INFECTIVE DISEASES,
from 2,227 in 1875, to 1,723 in 1876, to 951 in 1877, to 698 in
1878, to 157 in 1879, and to 48 in 1880.
In England the Pleuro-pneumonia Act came into force on
September Ist, 1890. Notification was to be given by the owner
to a police constable of the district, who was required to transmit.
the information to the Local Authority and also to the Board of
Agriculture. An inspector, with the aid of the veterinary surgeon,
arranged for the slaughter of the suspected animal, and, if the
disease proved to be pleuro-pneumonia, of the rest of the herd. The
results are shown in the following table :—
Diseased Cattle. Cattle
Number Number Number Healthy alauepter ee
VExns of of of Cattle in |e Ea?
"| Infected Fresh Cattle contact phan font
Counties, | Outbreaks. | Attacked. : B slaughtered. Pleuro-
Killed. Died. Pneumonia.
1890 36 465 2,057 2,022 37 11,301
1891 27 192 778 778 _ 9,491 232
1892 10 35 134 134 orl 3,477 188
1893 4 9 30 30 —_— 1,157 86
1894 2 2 15 15 —_— 391 41
Thus the number of cases was reduced from 2,057 in 1890 to
15 in 1894.
A departmental committee appointed in 1892 to inquire into
pleuro-pneumonia and tuberculosis, came to the following conclusions
with regard to pleuro-pneumonia :—
(1) That the system of compulsory slaughter be applied not only to.
all diseased cattle, but also to all cattle which have been in association
with them, or otherwise in any manner exposed to the infection of the
disease.
(2) Compulsory slaughter should be accompanied by supplementary
measures, such as restrictions on the movement and sale of cattle within,
or coming from, infected districts.
(3) Any exception to, or modification of, the system of compulsory
slaughter, as provided in the Slaughter Order, 1888, should only be
applicable to cattle in the dairy yards, byres, and cowsheds of large
towns, the owners or occupiers of which may claim in writing the privi-
lege of exemption for their cattle from immediate slaughter, on the
following conditions :—
INFECTIOUS PLEURO-PNEUMONIA. 245
(a) No head of cattle that has been brought into such dairy premises
shall be removed therefrom, except for the purpose of immediate
slaughter.
(b) In the event of an outbreak of pleuro-pneumonia, all the diseased
cattle shall be slaughtered.
(c) All the remaining cattle on such premises where an outbreak has
occurred shall be branded, and regularly subjected to the ther-
mometer test ; and whenever a continuous increase of temperature, ;
rising above 104°, is shown, they shall be slaughtered.
(d) No fresh cattle shall be admitted into such premises while any
of the cattle thus branded remain alive.
(4) Inoculation cannot be recommended as a means of eradicating
pleuro-pneumonia, nor as practicable under existing conditions. Although
it is open to owners to inoculate their cattle, it should be distinctly
understood that that operation shall not give them any immunity from
the regulations above suggested.
The order at present in force is the Pleuro-Pneumonia Order
of 1895. In addition to regulations for the movement of cattle,
for disposal of carcasses, for markets, and for compensation for
slaughter, the Order contains the following provisions :—
NotiFIcaTIon.
Notice of Disease.
(1) Every person having or having had in his possession or under his
charge a head of cattle affected with or suspected of pleuro-pneumonia
shall with all practicable speed give notice of the fact of the head of cattle
being so affected or suspected to a constable of the police force for the
police area wherein the head of cattle so affected or suspected is or was.
(2) The constable receiving such notice shall immediately transmit
the information by telegraph to the Board of Agriculture.
(3) The constable shall also forthwith give information of the receipt
by him of the notice to an Inspector of the Local Authority, who shall
forthwith report the same to the Local Authority.
Pa
Duty of Inspector to act immediately.
(1) An Inspector of a Local Authority on receiving in any manner
whatsoever information of the supposed existence of pleuro-pneumonia,
or having reasonable ground to suspect the existence of pleuro-pueumonia,
shall proceed with all practicable speed to the place where such disease,
according to the information received by him, exists, or is suspected to
exist, and shall there and elsewhere put in force and discharge the powers
and duties conferred and imposed on him as Inspector by or under the
Act of 1894 and this Order.
(2) The Inspector shall forthwith report to the Board of Agriculture.
246 -INFECTIVE DISEASES.
No Movement into or out of Plewro-pnewmonia Infected Place without
Licence.
Cattle shall not be moved into or out of an Infected Place except with
a Movement Licence of an Inspector or officer of the Board, and such
cattle shall not be moved except in accordance with the conditions
contained in such Licence.,
Pleuro-pneumonia found in Market, Railway Station, Grazing Park,
or other like Place, or during Transit.
The Inspector of the Local Authority shall cause to be seized all the
cattle affected with pleuro-pneumonia, and also all cattle being in or on
the market, fair, sale-yard, place of exhibition, lair, landing-place, wharf,
railway station, common, uninclosed land, farm, field, yard, shed, park, or
other such place as aforesaid, and shall forthwith transmit the information
by telegraph to the Board of Agriculture.
The Inspector of the Local Authority shall cause all such cattle so
seized to be detained at the place where they are seized, or to be moved
to some convenient and isolated place and there detained.
Removal of Dung or other Things.
Tt shall not be lawful for any person to send or carry, or cause to be
sent or carried, on a railway, canal, river, or inland navigation, or in a
coasting vessel, or on a highway or thoroughfare, any dung, fodder, or
litter that has been in an Infected Place, or that has been in any place in
contact with or used about a diseased or suspected head of cattle, except
with a Licence of an Inspector or officer of the Board or of an Inspector
of the. Local Authority.
Report to Board of Cattle that have been in Contact with Cattle affected
with Pleuro-pneumonia.
Where it appears toa Local Authority that there is within their District
any head of cattle which bas been in the same field, shed, or other place,
or in the same herd, or otherwise in contact, with any head of cattle
affected with pleuro-pneumonia, or otherwise exposed to the infection
thereof, the Local Authority shall forthwith report the facts of the case
to the Board of Agriculture.
Disinfection.
An Inspector or officer of the Board may cause or require any shed or
other place which has been used for a head of cattle while affected with
or suspected of pleuro-pneumonia, and any utensil, pen, hurdle, or other
thing used for or about such head of cattle, to be cleansed and disinfected
to his satisfaction.
Occupiers to give Facilities for Cleansing.
*q) The owner and occupier and person in charge of any shed or other
place which has been used for any head of cattle while affected with or
suspected of pleuro-pneumonia shall give all reasonable facilities to an
INFLUENZA. 247
Inspector or officer of the Board for the cleansing and disinfection of such
place, and of any utensils, pens, hurdles, or other things used for or about.
such cattle.
(2) Any person failing to comply with the provisions of this Article
shall be deemed guilty of an offence against the Act of 1894.
Prohibition to Expose or Move Diseased or Suspected Cattle.
(1) It shall not be lawful for any person—
(a) To expose a diseased or suspected head of cattle in a market or
fair, or in a sale-yard, or other public or private place where cattle
are commonly exposed for sale ; or
(b) To place a diseased or suspected head of cattle in a lair or other
place adjacent to or connected with a market or a fair, or where
cattle are commonly placed before exposure for sale ; or
(c) To send or carry, or cause to be sent or carried, a diseased or
suspected head of cattle on a railway, canal, river, or inland
navigation, or in a coasting vessel; or
(d) To carry, lead, or drive, or cause to be carried, led, or driven,
a diseased or suspected head of cattle on a highway or thorough-
fare : or
(e) To place or keep a diseased or suspected head of cattle on common
or uninclosed land, or in a field or place insufficiently fenced, or
in a field adjoining a highway unless that field is so fenced or
situate that cattle therein cannot in any manner come in contact
with cattle passing along that highway or grazing on the sides
thereof ; or
(f) To graze a diseased or suspected head of cattle on pasture being
on the sides of a highway ; or
(g) To allow a diseased or suspected head of cattle to stray on a
highway or thoroughfare or on the sides thereof or on common
or uninclosed land, or in a field or place insufficiently fenced.
INFLUENZA.
Influenza is an infectious disease characterised by a catarrh of
the respiratory or the gastric mucous membrane, accompanied by
great prostration and mental depression, and frequently ending
fatally by pneumonic complication, One attack is not protective.
The disease has occurred in the form of great epidemics, like the
pandemic of 1890, which is said to have started from Bokhara, and
travelled to St. Petersburg, Berlin, Paris, and London, whence it
spread all over this country. The incubation period is extremely
short, only a few hours, so that numbers are attacked almost
simultaneously. The occurrence of cases in succession in a family,
the importation of the disease by an infected person, and the escape
of persons in completely isolated localities, point to the existence
of a living contagium. Pfeiffer claims to have identified it with a
248 INFECTIVE DISEASES.
bacillus which was found by him in the purulent bronchial secretion,
and, by Canon, in the blood.
Bacillus of Influenza.—Very small rods, singly or in leptothrix
filaments. They stain with the aniline dyes, but not by Gram’s
method. They are non-motile and aerobic; they do not grow in
gelatine at the temperature of the room. On glycerine-agar very
small transparent drop-like colonies develop in about twenty-four
hours. In broth there is only a very scanty growth of whitish
particles on the surface, which subside and form a woolly deposit.
They are found especially in the bronchial -secretion, and only in
cases of influenza. Canon obtained them by puncturing the finger,
Fic. 121.—Baciiius or INFLuenza.
From a culture on gelatine, x 1000. (Itzerorr anp NIEMANN.)
and allowing a few drops of the blood to fall upon the surface of
glycerine-agar in a Petri’s dish, The organism will retain its
vitality for fourteen days in sputum, but is quickly detroyed by
drying. It is said that by applying the bacillus to the nasal
mucous membrane in monkeys, symptoms similar to influenza were
produced.
METHOD oF SraINING.
To stain the bacilli use Neelsen’s solution or Liffler’s methylene-
blue; or the following method :—
Canon’s method.—Spread the blood on cover-glasses, allow them
to dry, immerse for five minutes in absolute alcohol, and stain in the
following solution :—
INFLUENZA. 249
Aqueous solution of netbytone rind ong 40 parts; 3 per
cent. solution of eosin in 70 per cent. aldohol, 20 parts; distilled
water, 40 parts.
Float the cover-glasses from three to six hours in a capsule
placed in the incubator at 37° C., wash with water, and dry and
mount in balsam. The red corpuscles will be stained pink, and
the leucocytes, with the bacilli in them, blue.
/ / ¢
aoe ad 7
au” . Pete
‘ er $ ~ t
s eo a ‘
Sweet H SON
8 Sus: %
o Ne \ Se
, shee Ne
v4 Coad
gee
?
_ Fig. 122.—BaciLtus or INFLUENZA.
From a cultivation showing filaments composed of long and short rods,
cocci-forms and irregular elements. x 1200.
Equine INFLUENZA.
Equine influenza, or “ pink-eye,” has been noticed to be prevalent
at the same time as epidemics of influenza in man, but there does
not appear to be any evidence of intercommunicability or of any
relation between the two diseases.
CHAPTER XVIII.
ORIENTAL PLAGUE,—RELAPSING FEVER.—TYPHUS FEVER.—YELLOW
FEVER.
THE PLAGuE.
THe plague is a highly infectious disease, having its origin in
putrefaction and filth, in tropical climates. The virus in its effects
resembles that of typhus. The period of incubation varies from a
few hours to a week. The disease produces high temperature and
decomposition of the blood, and dark hemorrhagic patches appear
on the skin, but there is no eruption. Lymphatic inflammation and
buboes almost invariably occur. The virus is intensified by warmth
and overcrowding in houses, and dissipated by exposure to fresh air.
When the plague occurred in this country it was recognised as a
foreign pestilence from the East, and once imported it was fostered
and intensified in virulence wherever there was filth, putrefaction,
and overcrowding. The disease, like the small-pox, was communicated
from one person to another. If a case occurred in a house other
inmates were liable to suffer from the disease, while visitors to the
house ran a similar but less risk. There was a good deal of variation
both in the infectivity of the virus and in the susceptibility of
individuals, so that one contemporary writer remarked that “no
one can account for how it comes to pass that some persons shall
receive the infection and others not.”
Medical men were credited with enjoying an extraordinary degree
of immunity, though there were members of the medical profession
who undoubtedly died of the plague. This tradition has been
supported, to a certain extent, by the experience of the plague
in modern times. In the epidemic in Egypt, in 1835, of the ten
French physicians engaged there, only one died; and while those
who buried the victims of the plague were liable to suffer from
it, and many did so, yet the medical men made more than one
hundred post-mortem examinations without any death resulting.
250
THE PLAGUE. 251
The clothes and coverings of the infected often spread the disease,
and yet there are numerous examples of persons who without having
adopted any method of protection occupied the beds of plague patients
without contracting the malady.
The plague is transmissible from one country to another
by sea. An infected ship becomes an infective centre as readily
as an infected house. Once imported, whether by land or sea, the
virus from infected persons or merchandise spreads wherever the
environment is favourable for its development and extension.
Old London afforded in every way a suitable environment for
the plague. The situation of the city was unhealthy, and the old
town ditch was a receptacle for all kinds of filth. The houses
projected over the roadway, and the streets were saturated with
constant contributions of slops and of excrement from animals and
human beings. The houses were often filthy and unventilated, and
the floors strewn with rushes, which were seldom changed. Erasmus
goes so far as to say that the rushes were piled the new upon the
old for twenty years, and were fouled with spillings of beer,
fragments of fish, expectoration, vomit, excrement, and urine.
Another very striking insanitary feature of Old London was the
overcrowded state of the graveyards, which was well calculated to
predispose to pestilence, if not actually to produce it. The burials
were so frequent in St. Paul’s Churchyard that a new grave could
scarcely be dug without bodies being exposed in all stages of
putrefaction.
In 1894 the plague broke out in China, with all the symptoms
of the fatal bubonic pest of Old London. The disease was confined
to the poorest classes and the most overcrowded and most filthy
localities. In Canton the deaths exceeded one hundred thousand,
and in Hong-Kong numbered about ten thousand. The disease
was contagious, and mainly diffused by personal contact. Death
occurred, as a rule, in from twenty-four hours to five days. The
English and European community escaped, with the exception of a
very few out of a large number, mostly soldiers, employed in cleansing
the houses. The disease was a specific fever, intensely fatal, accom-
panied by high temperature, cerebral congestion, delirium, and the
formation of buboes. The buboes consisted of exquisitely painful and
swollen lymphatic glands. All the glands, in some cases, were affected.
According to Cantlie the glandular swelling when first recognised
was almond-shaped in the inguinal region, and globular in other
regions, with peri-glandular edema. The swelling rapidly increased
in size, becoming softer, less definite in outline, and less tender, until
252 INFECTIVE DISEASES.
by the end of five or six days it consisted of an elevated mass, doughy
to the touch, almost circular, with a diameter of six inches. The
skin over the swelling was livid and dimpled. The swelling was in
some cases due to purulent effusion, but more frequently on incision
there was only an escape of sero-sanguineous fluid. The cervical
glands in very severe cases sometimes attained an enormous size.
Three out of seven Japanese medical men were attacked and one
died, but none out of eleven English doctors, though they were equally
exposed toinfection. Of eight Englishmen attacked seven were among
the soldiers employed, and only two died. No nurses or attendants
on the sick were attacked. The virus appeared to be intimately
connected with filth in the soil. According to the Chinese, rats,
poultry, goats, sheep, cows, and buffaloes are susceptible. In the
houses and hotels dead rats were found in great numbers : it was said
that they emerged from their haunts in sewers and drains, appeared
to be dazed, and limped about, owing to the formation of buboes in
their hind legs. Rats, mice, and guinea-pigs inoculated with virus
from a human lymphatic gland died with development of buboes.
It appears to be clearly proved that rats suffer from the plague in
common with man, and it has also been suggested that they may
serve to spread the disease. Bacilli were found in human blood and
in the swollen lymphatic glands by Kitasato, and independently by
Yersin.
Bacillus of Plague.—Short rods with rounded ends. They
stain with aniline dyes, but not by Gram’s method. The stain
collects at the ends of the rods, leaving a clear space in the middle.
Sometimes the rods are surrounded by a capsule. They are found
in abundance in the buboes, and in small numbers in the blood in
very serious and rapidly fatal cases.
Material from the buboes inoculated on agar gives rise to white
transparent colonies, which have an iridescent edge when. examined
by reflected light.
The bacilli grow more readily on glycerine-agar and on solidified
serum. In broth cultures the liquid remains clear, and a flocculent
deposit forms on the sides and at the bottom of the vessel.
An alkaline solution of peptone 2 per cent., with from 1 to 2 per
cent. of gelatine, is the best nutrient medium. In cultures the
bacilli develop chains of short rods and well-marked involution
forms. Swollen and degenerated forms are found most abundantly
in old cultures, and stain with difficulty.
Mice, rats, and guinea-pigs, inoculated with bubonic tissue, die in
a few days, numerous bacilli being found in the lymphatic glands,
THE PLAGUE. 253
spleen, and blood. Guinea-pigs die in from two to five days, and
mice in one to three days.
In guinea-pigs after some hours there is cedema at the seat of
inoculation, and the lymphatic glands are swollen. After twenty-
four hours the animal refuses to eat, has a staring coat, and after
a time suddenly falls on its side, and is attacked by convulsions,
which become more and more frequent until death occurs.
After death the seat of inoculation is found to be extensively
edematous, and the neighbouring lymphatic glands enlarged and
filled with bacilli, The intestine is often congested, and the liver is
Fic. 123.—Bacitui or PLacuE aND PHacocyTeEs, x 800.
From human lymphatic gland. (Aoyama.)
congested and enlarged. In less acute cases an abscess of the
abdominal wall occasionally results.
The bacilli are sometimes found in the pleural and peritoneal
exudation. The liver and spleen also contain many bacilli. Those
in the blood are a little longer than those in the lymphatic glands.
Inoculations can readily be made from guinea-pig to guinea-
pig by using the pulp of the spleen, or the blood. Cultures lose their
virulence gradually, but the virus can be intensified by successive
inoculations in animals. The disease is infectious to mice as
well as inoculable. Pigeons are insusceptible, Rats and flies may
convey the bacilli.
According to Aoyama, the bacilli found in the blood of plague
254 INFECTIVE DISEASES.
patients and in the buboes are not identical. The bacilli in the
buboes are different in form, and they stain by Gram’s method.
There is no doubt that the micro-organism which was found in
blood is very similar to the bacillus of fowl cholera, and it is quite
possible that the so-called plague bacillus is really identical with the
bacillus of hemorrhagic septicemia, and that the real nature of the
contagium in bubonic plague is unknown.
Protective Inoculation.—Yersin, Calmette, and Borrell claim
not only to have produced immunity, but to have cured animals
after infection. Cultures on agar heated to 58° C. for an hour
were attenuated, and rabbits after intravenous or subcutaneous
inoculation were protected against virulent cultures. The serum
of immunised rabbits was capable of protecting from subsequent
virulent cultures, and neutralising the effect of a previous inoculation
of a virulent culture. A horse was inoculated with cultures which
killed mice in two days, and after six weeks a serum was obtained
which produced immunity in mice and guinea-pigs.
Stamping-out System.—It is not until the sixteenth century
that we hear of preventive measures being attempted in England,
and then they appear to have been adopted only when an outbreak
threatened to be very serious.
Early in the sixteenth century all those who had the plague in their
houses were ordered to put up wisps, and to carry white rods in their
hands.
In 1543 the Plague Order of Henry VIII. was issued. In place of
wisps the sign of the cross was to be made on every infected house, and
to remain there for forty days. Persons afflicted with the disease were
to refrain, if possible, from going out of doors, or for forty days to carry
a white rod in the hand. All straw from the infected houses was to be
carried into the fields and burnt. Churchwardens were directed to keep
beggars out of churches on holy days, and all streets and lanes were to
be cleansed.
In 1547 the means of notification was a blue cross with the addition
of the inscription Lord have mercy upon us. Later on the colour of the
cross was changed to red.
With the outburst of the plague in 1563, came an attempt to enforce
a terrible system of compulsorily shutting up infected families. The
doors and windows in such houses were to be closed, and no inmates were
to leave the premises and no visitors to be allowed for forty days. No
better incubator on a large scale could possibly have been devised for
both breeding and intensifying the virulence of the plague bacillus, or
whatever may be the contagium vivum of this disease.
This compulsory shutting up of the sick with the healthy amounted
to a compulsory infection of many of the unfortunate inmates who might
THE PLAGUE. 255
otherwise have escaped, and very naturally the order was frequently
infringed.
In 1568 the Lord Mayor of London drew up instructions for the
Aldermen for dealing with the plague. It was enacted that constables
and officers should search out infected houses and report to the authorities,
In other words, that there should be notification by the police. All infected
houses were to be shut up, and no person to be allowed to come out for
twenty days, All bedding and clothes used by the victims were to be
destroyed.
At Westminster these instructions were to be enforced under a penalty
of seven days in the stocks, with imprisonment to follow, making in all
a punishment of forty days.
In 1581 the Lord Mayor transferred notification from the constables
to searchers. Two honest and discreet matrons in every parish were to
search the body of every such person that happened to die in the parish.
They were ordered to make a true report to the clerk of the parish, and
the said clerk had to report to the wardens of the parish. For failing
to notify, the penalty was an exemplary term of imprisonment. The
searchers were of course likely to be offered heavy bribes by the people
to suppress reports, owing to their anxiety to avoid the shutting up of
infected houses. i
The continued prevalence of the plague led to the publication, in
1593, of a book by Simon Kellwaye. One chapter ‘‘teacheth what
orders magistrates and rulers of citties and towns should cause to be
observed,” which included among other regulations that no dunghills
were to be allowed near the city, and the streets were to be watered
and cleansed. F
No surgeons or barbers who let blood were to cast the’ same into the
streets. All those visiting and attending the sick to carry something in
their hand to be known from other people ; and if the infection were in
few places, all the people were to be kept in their houses during the time
of their visitation, and when this was over, all clothes, bedding, and
other such things used upon the sick, were to be burnt.
In 1603 Thomas Lodge recommended that discreet and skilful men
should be appointed in every parish to notify sickness to the authorities,
and so cause them to be visited by expert physicians, and that such as
were sick should be separated from the whole, and that isolation hospitals
should be built outside the City in separate and unfrequented places.
In 1665 the Great Plague of London occurred, and was attributed by
some to the importation of an infected bale of silks from the Levant.
According to Hodges the disease stayed among the common people, and
hence was called The Poor's Plague. He criticised the system of shutting
up infected houses, and strongly recommended that those who were
untouched in infected houses should receive “ accommodation outside the
city.” The sick were to be removed to convenient apartments provided
on purpose for them. To quote his own words, “ Timely separation of
the infected from the well is absolutely necessary to be done.”
For the purification of houses his directions were to place “a chafing
256 INFECTIVE DISEASES,
dish in the middle of the room, where proper things were burnt and
exbaled all around.” The use of sulphur and quicklime was mentioned. *
Preventive measures were drawn up and published by the Lord Mayor
and Aldermen. Examiners in Health, watchmen, and searchers were
appointed. Surgeons were selected to assist the searchers in making their
reports, and a fee of twelve pence was allowed for every case. The disease
was immediately notified to the Examiner of Health. Rules for disinfec-
tion were made, and every infected house was shut up, and no one removed
except to a pest-house or tent. Orders were issued for cleaning and sweep-
ing the streets. Hackney coaches were not to be used after conveying
patients to the pest-house until they had been well aired, Regulations
were also made dealing with loose persons, assemblies, and drinking taverns.
The plague was scarcely over before the whole city was in flames. A
new city speedily rose upon the ashes of Old London. A few sporadic
cases of plague are given in the London Bills of Mortality down to 1679,
when they finally ceased. London was sterilised by the great fire. “ Great
as this calamity was,” wrote Thomas Pennant, ‘yet it proved the provi-
dential cause of putting a stop to one of far more tremendous nature.
The plague, which, for a series of ages, had, with very short intervals,
visited our capital in its most dreadful forms, never appeared there again
after the rebuilding of the city in a more open and airy manner ; which
removed several nuisances, which if not the origin of a plague, was
assuredly one great pabulum, when it had seized our streets.”
In the years 1720-22 there was a terrible outburst of plague in France.
It was attributed at Marseilles to importation by a ship from Syria. This
caused a panic in England, and the Lords Justices considered it necessary
for the public safety that measures should be taken to defend the country
from a fresh invasion of this disease. Dr. Richard Mead was entrusted
with drawing up the required recommendations. Mead laid it down as
an essential doctrine that the plague was not native to this country, and
therefore the first thing was to prevent importation, and if such a misfor-
tune occurred, it was to be prevented from spreading. How was this to
be accomplished? Briefly stated, his system was as follows : Lazarettoes
were to be provided for the reception of infected men and merchandise.
The healthy were to change their clothes and to be kept in quarantine, and
the sick were to be kept remote from the healthy and their clothes.
destroyed,
If, through a miscarriage in the public care, by the neglect of officers
or otherwise, the disease was imported, then “the civil magistrates were
to make it as much for the interest of the afflicted families to discover:
* During outbreaks of the plague amulets were extremely popular. Walnuts
filled with mercury, pieces of cloth coated with arsenic, and arsenical cakes,.
were very generally worn. The College of Physicians recommended issues on
the arms and legs. Dr. Hodges wrote, that the more of the ulcers that were
made the better, although their largeness answered as well as more in number.
If two issues were preferred, it was recommended to make one on the left arm
and the other on the opposite leg. A somewhat similar plan was adopted in
Circassia by small-pox inoculators.
RELAPSING FEVER. 257
their misfortune, as it was when a house was on fire, to call in the
assistance of the neighbourhood.” ‘The shutting up of infected houses
was condemned in the strongest terms, and a system of notification and
isolation was proposed on the lines originally suggested by Dr. Hodges.
1. A Council of Health was to be established, and entrusted with such
powers as might enable them to see all their orders executed with im-
partial justice.
2. Notificution.—The ignorant old women employed as searchers were
to be replaced by understanding and diligent men, who were to report
cases immediately to the Council of Health.
3. Isolation.—Physicians were at once to be despatched to visit the
suspected cases, and when the suspicion of plague was confirmed, all the
families in which the sickness occurred were to be isolated. The sick
were to be separated from the sound, and isolation houses to be provided
three or four miles out of the town.
The removal of the sick was to be made at night, so as to avoid the
danger of spreading infection, and all possible care was to be taken to
provide such means of conveyance for the sick that they might receive no
injury. The poor were to be isolated in houses provided for the purpose,
but the rich were to be allowed to be in their own homes provided that
care was taken to separate the healthy from the sick, and no pains were
to be spared to provide clean and airy apartments. All expenses were to
be paid by the public, and a reward was to be given to the person who
made the first discovery of infection in any place.
Mead further pointed out that general sanitation must be carefully
attended to. Officers were to see that the streets were washed and kept
clean from filth, carrion, and all manner of nuisances. Beggars and idle
persons were to be taken up, and such miserable objects as were fit
neither for the hospitals nor for the workhouses, were to be provided for
in an establishment for incurables. Houses also were to be kept clean,
and sulphur was to be used as a disinfectant.
After centuries of experience we have learnt that the necessary
conditions for avoiding the plague are more accurate knowledge on
the part of the profession and the public of the way in which the
disease spreads, and the adoption of sanitary precautions, which must
include personal cleanliness, sanitary dwellings, absence of overcrowd-
ing, immediate notification, prompt separation of the sick from the
healthy, disinfection of infected dwellings, destruction of infected
clothing, and extra-mural burial or, better still, cremation. It was
because the very reverse of these sanitary conditions existed that
the virus of the plague found a suitable environment in Old London
and in recent times in Hong-Kong.
Revapsina FEvER.
Relapsing or famine fever is a contagious disease producing a
state of high fever lasting about seven days, followed by apparent
17
258 INFECTIVE DISEASES.
recovery, and in about fourteen days by another attack of fever,
which may be repeated after another week.
Starvation, in association with overcrowding and filth, is intimately
connected with the causation of the disease. The subjects of -the
disease contaminate the air around them, and the virus is principally
conveyed by tramps and dirty people.
Obermeier discovered spirilla in the blood during the paroxysms
of fever. The constant occurrence of the spirillum in relapsing
fever, and the fact of its not being found in any other conditions,
render it very probable that it is the cause of the disease.
Fic. 124.—Spritium OBERMEIERI IN BLoop or Monxkry INocULATED WITH
SPIRILLA AFTER REMOVAL OF THE SPLEEN (SOUDAKEWITCH).
Spirillum Obermeieri (Spirocheta Obermeieri, Cohn).—
Threads similar to the Spirillum plicatile. In length they are mostly
16 to 40 mw, with regular screw-curves. They move very rapidly,
and exhibit peculiar wave-like undulations. They are absent from
the blood during the non-febrile intervals, but are found in the
interior of leucocytes in the spleen. In blood serum and 50 per
cent. salt solution, they preserve their movements. In cover-glass
preparations they are readily stained by any of the aniline dyes,
and in sections, by preference, with Bismarck brown. They are not
TYPHUS FEVER.—YELLOW FEVER. 259
found in the urine, sweat, or saliva. They have not been cultivated
artificially on any nutrient media. Monkeys have been successfully
inoculated with blood containing the spirilla by Koch, Carter and
Soudakewitch ; and Koch found the spirilla in the vessels of the
brain, liver, and kidneys, after death. According to Soudakewitch
a fatal result is produced in monkeys if the spléen is removed,
and the spirilla are found in great numbers in the blood; but if
the spleen is not excised the spirilla rapidly disappear, and recovery
follows. Miinch and Motschutkowsky transferred blood containing
the spirilla to healthy persons, and produced typical relapsing fever.
TypHus Fever.
Typhus fever is a highly contagious disease, which lasts for two
or three weeks, and produces a measly eruption. Like the plague,
it is intimately associated with overcrowding and filth, and is liable to
occur where these conditions exist in cities, in armies, and in prisons.
The virus produces profound changes in the blood, and after death
the internal organs are found to be congested, especially the lungs,
which are very friable. The spleen is softened and often enlarged,
and the blood is dark and imperfectly coagulated.
The virus is dissipated by fresh air. It is given off by the
breath of patients, and possibly from the skin. It clings to the
clothes of patients, and the disease may be conveyed by their agency.
One attack, as a rule, confers immunity. Some persons are naturally
insusceptible, failing to contract the disease though daily exposed to
it. Hlava has described a bacterium which he believes to be the
specific micro-organism, Thoinot and Calmette found the same
bacterium with others, but there was no particular micro-organism
constantly present. There can be little’ doubt that the nature of
the contagium is unknown.
Stamping-out System.—Sanitary precautions, and especially
the operation of the Public Health Acts in relation to lodging-
houses, prisons, and the better housing of the working classes, have
been instrumental in almost completely stamping out the disease in
this country.
YELLOW FEVER.
Yellow fever is a disease of tropical climates, characterised by
abdominal tenderness, hemorrhagic vomiting (black-vomit), and
jaundice. The disease may end fatally, or recovery occur in about
two or three weeks. It is especially prevalent in the West Indies
and in parts of North and South America.
260 INFECTIVE DISEASES.
The virus may be conveyed by infected ships, and has in this
way made its appearance at British and French seaport towns.
The disease is generally believed to be contagious, but the source
of the virus is not known. According to Sternberg the virus is not
conveyed by water, but spreads where there is overcrowding and
filth.
Bacteria in Yellow Fever.—Freire asserts that there is a
specific micrococeus in yellow fever which can be grown on all
ordinary nutrient media, and that cultures can be used for protective
inoculation with satisfactory results. Carmonay Valle also claims to
have discovered the contagium ; but Sternberg, who has carried ‘on
investigations extending over several years, maintains that there is no
characteristic micro-organism present in the blood or in the tissues
after death. Aerobic and anaerobic cultures were made from the
blood, liver, kidney, urine, stomach, and intestines. The liver was
found to contain after death a number of bacilli, most frequently
Bacillus coli communis and Bacillus cadaveris. Blood or fresh
liver does not produce any disease in rabbits or guinea-pigs, but
liver tissue kept for forty-eight hours and inoculated subcutaneously
in guinea-pigs is extremely pathogenic. Similar results occur after
inoculation of healthy liver which has been kept in the same way.
We may conclude from these experiments that the nature of the
contagium is unknown.
Stamping-out System.—Sternberg states that there are many
facts relating to the origin and extension of yellow fever epidemics
which support the theory that the virus is present in the evacua-
tions, and that accumulations of fecal matter and of organic
material of animal origin furnish in certain climates a suitable soil
for the development of the contagium. According to this view the
evacuations should be thoroughly disinfected, and with other sanitary
precautions and efficient quarantine at seaports, the disease may
be stamped out, and the danger of importation from the natural
home of the disease reduced to a minimum,
CHAPTER XIX.
SCARLET FEVER.—MEASLES.
ScaRLer FEvEr.
ScarLet Fever is a highly contagious disease peculiar to man. It
produces inflammation of the tonsils and adjoining parts, fever, and
a general punctiform eruption. The period of incubation is about,
a week, and the rash usually appears on the second day. In some
cases the disease manifests itself in an extremely mild form, known
as latent scarlet fever, in which there is only a slight febrile attack,
or a mild sore throat, with very little or no rash. Many cases
would not be recognisable as such if they were not capable of
conveying scarlet fever, or unless other cases followed or occurred
simultaneously which were undoubtedly typical cases of the disease.
The occurrence of such cases in the early history of an epidemic
often causes the greatest difficulty in tracing the origin of the
outbreak, and indeed in some cases renders it quite impossible to
do so.
The virus is given off by the skin, in desquamation, and possibly
by the urine. It maintains its vitality in clothing for months, and
sometimes longer. It may also be conveyed by the hands of the
physician to women during parturition. The disease may be
transferred by subcutaneously inoculating persons, who have not
previously contracted scarlet fever, with virus obtained by puncturing
the eruption on the skin.
‘After death the internal organs appear to the naked eye more or
less healthy. The liver is soft, the kidneys are congested, the ileum
is inflamed, and Peyer’s patches enlarged and congested; but these
conditions are also produced by other causes. There are inflam-
matory changes in the lymphatic follicles of the tonsils, and the
larynx and trachea. Other morbid lesions, especially in the kidneys,
are associated with the sequele and complications, and though
commonly occurring in scarlet fever are also found in other diseases.
261
262 INFECTIVE DISEASES.
These changes appear to be due to the poison which is in the blood,
and is excreted by the kidneys. The epithelium is in a state of
cloudy swelling, a condition found in other febrile diseases and in
septic poisoning.
Bacteria in Scarlet Fever.—The occurrence of micro-organ-
isms in cases of scarlet fever has been observed by several investi-
gators—Coze and Feltz, Crooke, Léfiler, Babés, Heubner and Bahrdt,
and notably by Frankel and Freudenberg, and more recently by
Klein, the author, Raskin, and others.
Coze and Feltz found cocci in the blood, and Crooke, in cases of
scarlet fever with severely affected throat, found bacilli, cocci, and
streptococci in the organs of the throat, and cocci in the internal
organs. Crooke left it an open question whether these cocci were
the specific organisms of scarlet fever, or were to be regarded as
diphtheritic or septic associates. He inclined, for clinical reasons,
to the latter view.
Loffler, in cases of scarlatinal diphtheria, found the same chain-
forming micrococeus which he had found in typical diphtheria.
Babés was able constantly to prove the presence of « strepto-
coccus in inflammatory products secondary to scarlatina.
Heubner and Bahrdt, in a fatal case of scarlet fever in a boy,
complicated with suppuration of the finger and knee-joints, and
with pericarditis, found a streptococcus identical in form with Strepto-
coceus pyogenes, but cultivations were not made. The secondary
infection started from diphtheritically affected tonsils, which were
followed by retro-pharyngeal abscesses.
Frinkel and Freudenberg examined, for micro-organisms, three
cases of scarlatina with well-marked affection of the throat. In all
three cases they obtained cultivations of cocci from the submaxillary
lymphatic glands, spleen, liver, and kidney. These cocci could not
be distinguished from Streptococcus pyogenes derived from pus, nor
from the undoubtedly identical streptococcus which one of them
(A. Frankel) had repeatedly cultivated in large numbers from
puerperal affections. In two of the cases Streptococcus pyogenes
was the only organism present, and in all three cases it was far in
excess of other colonies which developed. The organisms were also
found in sections of the organs by microscopical examination.
Frankel and Freudenberg could in no way distinguish the strepto-
coccus in scarlatina from the streptococcus in pyemia and septi-
cemia. The identity of this streptococcus with Streptococcus pyogenes
and Streptococcus puerperalis was established by comparison of their
macroscopical and microscopical appearances in cultivations on
SCARLET FEVER. 263
nutrient agar-agar, nutrient gelatine, and in broth, both at the
ordinary and at higher temperatures, and also by experiments on
animals. They concluded that it could be stated with certainty
that the organisms in question did not stand in causal relation to
searlet fever. They considered that special methods of microscopical
and biological research were apparently needed for demonstrating
the true scarlet fever contagium, which probably was especially
present in the skin. They considered that the presence of the
—_
a. b. Cc.
Fic. 125.—Pure-CurtivaTions or STREPTOCOCCUS PYOGENES.
(a) On the surface of nutrient gelatine; (b) In the depth of nutrient gelatine ;
(c) On the surface of nutrient agar.
streptococcus was due to a secondary infection, to which the door
was opened by the lesions of the throat—a view which was sup-
ported by the fact that the organisms were found in submaxillary
lymphatic glands. They preferred to use the term “secondary ”
to “complicated” or “combined” infection, because this expresses
the fact that by the effect of the scarlatinal virus the soil is
rendered suitable for this ubiquitous microbe when it has once
gained an entrance.
This streptococcus was found by Klein in five out of eleven cases
264 INFECTIVE DISEASES.
of scarlet fever in man, twice in association with certain other
micro-organisms, and three times alone. The micro-organisms were
isolated by inoculating tubes of nutrient gelatine, solidified obliquely,
by streaking the surface with blood taken from the finger, the arm,
or the heart after death. Those cases from which the organism was
obtained were all cases with ulcerated throat, and the culture
experiments, from the living patient, were made on or about the day
at which the temperature was at its maximum,
e iti Source , icro-Organisms Death or
Name of Patient. Oe a aid. ca eolaiad: Recovery.
1) L— F—_. Severely | Finger Streptococcus Ultimately
aged 5 ulcerated ‘ recovered,
‘ Staphylococcus
2; K— jf aa Much eee aureus Died of
2 : u micro- Z
age ulcerated o me ee c pyemia.
Streptococcus
3) H—— L—. | Ulcerated ‘i Staphylococcus
aged 8 Uae oni Recovered.
4 (a woman), Much Arm None (Not stated.]
aged 40 ulcerated
5 (a girl), » s »
aged 19
6}; B— AT ae Ulcerated | Finger Streptococcus Recovered,
age
7; E——W—, Much a 4 None [Not stated.]
aged 22 ulcerated i
8| R—— H——, | Ulcerated ae od es ro
aged 8 i
9| F G—, re Heart | Streptococcus Died.
aged 23 |
10; ss _— ‘3 | None 4
aged 3
11 R— B——., Advanced 45 | F -
aged 20 months | ulceration |
Klein regards this streptococcus as the actual cause of scarlet
fever in man.
The author, Raskin, Holmes, and others who have investigated this
subject agree with the conclusions of Frankel and Freudenberg. The
author is convinced that the streptococci in suppuration, puerperal
septicemia, pyeemia, and septicemia, and in certain cases of measles,
searlatina, and diphtheria, are identical; and from overwhelming
evidence we are justified in concluding that—(1) The nature of
the contagium of scarlet fever is unknown. (2) The streptococcus
regarded by Klein as the contagium is the Streptococcus pyogenes.
MILK-SCARLATINA. 265
(8) This streptococcus is found, sometimes in company with Staphylo-
coccus pyogenes aureus, as a secondary result in scarlet fever and
many other diseases, and its exact relation to scarlet fever and its
identity with the streptococcus from pus and puerperal fever, were
definitely established in 1885 by Frankel and Freudenberg.
MiLK-Scaratina.
It would not be necessary to say anything further on the etiology
of scarlet fever if the generally accepted belief, that scarlet fever is
a disease peculiar to man, were accepted by the Medical Department
of the Local Government Board; but the theory is officially held
that scarlet fever is in its origin a disease of cows. Bovine scarlatina
is supposed to be an eruptive disease of the teats, and it is maintained
that the virus, by contaminating the milk, produces scarlet fever in
the human subject. As this theory is very naturally accepted by
many medical officers of health, and is mentioned in English medical
text-books, it will be necessary to discuss this question in considerable
detail, and especially as these opinions were promulgated in this
country with official support, and have since been proved to be
erroneous.
The theory of the origin of the exanthemata in diseases of the
lower animals is a very old one. The Arabians imagined that small-
pox arose from the camel. Jenner adopted a similar theory, and
expressed his belief that small-pox originated in the horse, being
generated by horses suffering with “greasy” hocks. Thus Jenner
wrote: “‘ May not accidental circumstances have again and again
arisen, still working new changes upon it, until it has acquired
the contagious and malignant form ‘under which we now com-
monly see it, making its devastations among us? and from a
consideration of the change which the infectious matter undergoes
from producing a disease in the cow, may we not conceive that
many contagious diseases now prevalent amongst us may owe their
present appearance, not to a simple, but a compound origin? For
example, is it difficult to imagine that measles, scarlet fever, ulcerated
sore throats, and spotted skin, all spring from the same source,
assuming some variety in their forms according to the nature of
their new combinations?” Baron informs us that this idea was
prevalent in Jenner’s mind as early as 1787. It is related that in
that year he accompanied his nephew, George Jenner, into a stable
to look at a horse with diseased heels, and, pointing to them, he
remarked : “ There is the source of small-pox. I have much to say
266 INFECTIVE DISEASES.
on that subject which I hope in due time to give to the world.”
And again in 1794, when writing in connection with this subject,
he adds: “ Domestication of animals has certainly provided a prolific
source of diseases among man.”
Jenner’s views were found to be incorrect, and it was shown by
Loy and others that the grease bears no relation to cow-pox, and
it is now known that Jenner mistook horse-pox for the disease
known as the grease. No one at the present day supports Jenner’s
theory of small-pox in man arising from any disease of the horse.
Indeed, the origin of small-pox from a disease of the horse was not
upheld even by Jenner’s pupil and nephew, Henry Jenner. The
latter promulgated the idea that small-pox originated from the cow.
Tle believed that small-pox, in fact, was cow-pox intensified in its
virulence by being passed through man. He thus expressed himself -
“Nor may it, perhaps, be too hypothetical to suppose that the
cow-pox may possibly be the small-pox in its original unadulterated
state, before it became contaminated by passing through the impure
and scrofulous habits of human constitutions.” The theory of the
origin, in animals, of human febrile diseases was, later, advocated by
Copland, who stated, firstly, that scarlet fever in man was originally
a disease of the horse, and that it formerly occurred, and had recently
occurred, epidemically as an epizootic among horses; secondly, that
it was communicated in comparatively modern times from horses to
man; thirdly, that it might be, and had been, communicated to the
dog. But this opinion has not been accepted, for the disease called
scarlatina in the horse is a non-infectious disease, generally attack-
ing but one or two horses in a large stud. It neither spreads by
contagion nor infection ; and Williams states that it is impossible to
transmit it from the horde to-any other animal, and that many cases
of the so-called scarlatina of the horse are in reality identical with
purpura.
The theory was again revived, but in another form, i has
been adopted by the Medical Degarkneut of the Local Government
Board. Owing to failure in tracing,'in some cases of milk scarla-
tina, the contamination of the milk from a human source, the
theory was started that in such eases the disease is derived from
the cow—that, in other words, there is a disease, scarlet fever
in the cow, which is responsible for outbreaks of scarlet fever in
man.
In 1882 an epidemic of scarlatina in St. Giles and St. Pancras
was investigated by Mr. W. H. Power for the Board. The disease
was distributed with a milk supply from a Surrey farm. In this case
MILK-SCARLATINA. 267
two facts were ascertained : the one, that a cow recently come into
milk had been suffering from some ailment from the time of her
parturition, of which loss of hair in patches was the most con-
spicuous manifestation ; the other, that there existed no discoverable
means by which the milk could have received infective quality from
the human subject.
Tn 1885 an outbreak of scarlet fever occurred in Marylebone in
connection with milk from a farm at Hendon, and again Power
failed to establish infection from any human source in any commonly
accepted way—such, for example, as handling of milk, or milk utensils,
by persons carrying scarlatina infection. But on examining the
cows with a view to ascertain any new condition pertaining to
them, it came to light during the inquiry that some of them,
which had recently been introduced from Derbyshire, were suffering
from a vesicular disease of the teats.
At this stage Klein became associated with Power in the
inquiry; and their belief in the existence of a disease among the
cows on the farm capable of producing scarlatina among human
consumers of the cow’s milk, became unreserved. Klein took away
with him samples of milk, contents of vesicles, and discharges
from ulcers, and afterwards two of the cows were purchased and
kept under observation.
Dr. Cameron of Hendon has given a detailed description of the
clinical history of this disease. He expressed his belief that it was
a specific disease capable of being communicated to healthy cows
by direct inoculation of the teats with virus conveyed by the milker
from a diseased animal.
The condition of the teats is described as follows: The teats
became enlarged, swollen to nearly twice their natural size, and
edematous. On handling them there was no feeling of induration.
Vesicles appeared on the swollen teats and upon the udder between
or near the teats. These varied in number from two to four on a
teat, and in size from a pea to a horsebean. The vesicle contained
a clear fluid. The vesicles were rubbed and broken in milking, and
left raw sores, sometimes red, in other cases pale in colour, with
raised, ulcerated edges. Sometimes a few accessory vesicles formed
around the margin of these ulcerated sores. After the rupture of
the vesicle a brown scab formed, which might remain attached for
five or six weeks, or fall off in ten days or a fortnight, a smaller
one forming afterwards. A thin, watery fluid exuded from under
the scab, and the sore ultimately healed.
Cameron examined the teats of several cows five or six weeks
268 INFECTIVE DISEASES.
after they were attacked. The scabs then varied in size from a
shilling to a florin; they were about one-eighth of an inch thick in
the centre, thinning off towards the edges.
Some of the cows were also suffering from an eruption on the
rump and hind quarter, consisting of patches of eczematous crusts.
When a crust was picked off, the hair came off with it, exposing a
raw, moist sore, the crusts and sores looking exactly like eczematous
scabs and sores; but this condition corresponds in description with
eczema, the result of ringworm which is very common in young
stock,
In addition to his own observations, Cameron obtained infor-
mation from the farmers, and others familiar with cows, who
thought they recognised in the disease at the farm one stage of a
disease which they were able to describe. Cameron thus gives an
account of what he and his informants together would regard as
a connected clinical history of the disease.
He did not see the earlier symptoms, and hence these were
of necessity learnt from other persons. The account, therefore,
of these symptoms was to be held liable to future correction or
modification.
Cameron stated that he learnt this disease was capable of
being communicated to milkers by inoculation with virus from the
vesicles on the teats, though the milkers on the Hendon farm
escaped. ‘“ A trusty informant received the virus into a recent
scratch on the forefinger while milking a diseased cow. General
weakness, malaise, and loss of appetite resulted, and after about
four or five days a vesicle or small blister appeared on the finger.
This broke, and several others formed on the back of the hand.
The whole hand and fingers became swollen and inflamed, the
inflammation extending in broad lines as far as the elbow. The
general disturbance lasted a fortnight.”
In the course of the inquiry, Cameron adds that it was
strongly asserted by several people, who examined the cows, that
they were suffering from cow-pox. He, however, dismissed the
diagnosis of cow-pox on the ground that no papule had been
observed or subsequent formation of pustule, areola, or pitting, and
because the vesicles were not umbilicated. These reasons given
for dismissing the diagnosis of cow-pox at Hendon were totally
inadequate ; a comparison having been made between the characters
of the eruption of vaccinia as it appears on an infant’s arm, instead
of the eruption of the natural or so-called spontaneous disease on
the teats of the cow.
MILK-SCARLATINA. 269
Klein stated that on the teats and udders of two cows whicli
he investigated there were several flat irregular ulcers, varying in
diameter from one-quarter to three-quarters of an inch. Some
ulcers were more or less circular, others extended in a longitudinal
direction on the teat. The ulcers were covered with a brownish or
reddish-brown scab. The animals looked thin, but not strikingly so.
In feeding capacity, milking power, and body temperature there
was nothing abnormal.
Four calves were inoculated in the corium of the groin and the
inside of the ear, with scrapings from the ulcers after removal of
the crusts. In one, which may be taken as an example of the result
obtained, there was vesiculation at the margin of the spot inoculated,
and in the centre the commencement of the formation of a crust.
On the seventh day each sore on the ear had enlarged to about
half an inch in breadth, and was covered in its whole extent by
a brownish crust. On the eighteenth day they had all healed up
and become converted into flat scars.
To search for micro-organisms, Klein removed the crust
from an ulcer on the teat, scraped off the most superficial layer,
squeezed the ulcer, and made cover-glass preparations. Tubes of
nutrient gelatine and nutrient agar-agar were also inoculated, and
a streptococcus was isolated which in morphological and cultural
characters agreed with those of Streptococcus pyogenes.
Two calves were inoculated in the groin with the cultivated
micro-organism. One calf died in twenty-seven days. At the
necropsy there were found peritonitis, and hemorrhagic spots on
the omentum; the liver, kidneys, and lungs were congested, and
there were petechiz under the pleura, and pericarditis. The second
calf was killed, and at the necropsy the lungs and kidneys were
congested, and there were hemorrhagic patches on the spleen.
In these cases, the post-mortem appearances and anatomical
features recalled to Klein the lesions of scarlatina. In the
kidney, for example, the cortex was congested, and there were
hemorrhages, glomerulo-nephritis, and granular or opaque swelling
of the epithelial cells and infiltration with round cells. From the
blood of the heart the streptococcus, which had been used in the
inoculation, was recovered. In view of this evidence it was con-
cluded that the streptococcus was the virus of the cow disease, and
‘that it produced in calves a disease very closely resembling that of
scarlatina in man.
Two of the cows selected from the Hendon farm were killed,
and it was observed in one that the lungs were congested, and
270 INFECTIVE DISEASES.
that there were numerous adhesions by recent soft lymph between
the lower lobes of the lung and the costal pleura. In the
liver there were several reddish streaks and patches. The spleen
and kidneys, with the exception of slight congestion, appeared
normal.
Sections of the kidney showed well-marked glomerulo-nephritis
and infiltration of the sheath of the cortical arterioles with numerous
round cells. The epithelium of the convoluted tubules was swollen,
opaque, and in many places disintegrating.
In the other cow there was great congestion of the lungs and
pleural adhesions ; the cortex of the kidney was congested, but its
medulla was pale.
On microscopic examination there was a good deal of round-
celled infiltration in the walls of the infundibula and bronchi in the
Inng, and round the arterioles in the kidney.
In sections of the ulcers on the teats, the corium was found to be
infiltrated throughout the whole extent of the ulcer with round
cells. In the superficial layers of the stratum Malpighi, close to
the stratum lucidum, as also in the stratum lucidum itself, there
were numerous cavities of different sizes. These cavities lay side by
side, the most superficial ones being covered by the stratum lucidum,
or extending between the layers of this stratum.
At the marginal parts the cavities, although placed side by side,
were well separated from one another by thicker or thinner
trabecule, composed of epithelium, while at or near the centre of
the ulcer these trabecule were destroyed, the cavities had become
confluent, and the covering layers of the cuticle having here also
given way, their contents extended on to the free surface of the
ulcer. In short, Klein states that all the anatomical details of
the distribution and arrangement of these cavities recalled vividly
the conditions observed in the vesicles of cow-pox. Yet as a result
of this investigation he concluded that the cow disease at Hendon
was bovine scarlatina, and that towards its study and supervision
every effort ought to be directed in order to check the spread of
scarlet fever in man.
As a result of this conclusion, the Board of Agriculture resolved
to have the whole subject fully investigated, and the author was
directed to study the bacteriology and micro-pathology of this disease
and to report thereon. Professor Axe investigated the origin of
the outbreak of the disease in the cows, and Professor M‘Fadyean
carried out an investigation into the possibility of inoculating cows
with the virus from cases of scarlet fever in man.
MILK-SCARLATINA. 271
Tur AvutHor’s INVESTIGATION.
An outbreak of an eruptive disease of the teats, alleged to be
identical with the so-called Hendon cow disease, was raging in some
farms in Wiltshire. In this case every facility was given by the
owner of the estate for a thorough investigation into the disease.
Not only were animals sent from the farm to London, but the
author was allowed to visit the farms, to inspect all the infected
animals, and to make every investigation, with the hearty co-
operation of the bailiff of the farms, and the voluntary assistance
of the head cowmen and those under them. Some of these cowmen
were unusually intelligent, while two had had experience of cows
for more than half a century. Thus, there was not only every
opportunity for studying the disease on the lines indicated by
Klein, but it was possible by repeated visits to the farms to enter
into the clinical history of the disease in the cow, to study very fully
the nature of the disease on the hands of the milkers, and to trace
the probable mode of its introduction on the estate, and the way in
which it spread from one part of the herd to another.
Two cows were sent to London with disease of the teats and of
the udder between the teats. On the right teats of one there were
numerous sores, covered with crusts varying in size and in thickness,
and generally fissured. In some they were flat, in others conical ;
some were with difficulty removed with forceps, others were readily
detached. The crusts varied in colour from reddish-brown to very
dark brown or almost black. On detaching or scraping « crust
there was a granulating and somewhat indurated base. On the
right anterior teat there were several ulcers, from which appa-
rently the thick crusts had been detached, and new scabs were
forming. On the left posterior teat there were unusually large,
dark brown, or blackish crusts, covering a very extensive area
of ulceration, extending over the whole of the lower third of the
teat.
In the other cow from Wiltshire there was the same disease on
the teats, but not in such a severe form. The sores were covered
with thick crusts, but though varying in size they were more
regular in form, and more circumscribed.
Having entirely removed the crusts from some of the ulcers, a
number of inoculations in nutrient gelatine and nutrient agar-agar
were made from the discharge, and cover-glass preparations were
made and stained in the ordinary way. Cultures were obtained of
the organisms commonly found in pus.
272 INFECTIVE DISEASES.
With the discharge and with scrapings from the ulcers two
calves were inoculated.
Of the two calves, one was inoculated by scarification in both
ears; the other, a small calf, was scarified in the left ear. Scrapings
from the ulcers were rubbed into the places thus prepared. In
addition, in the small calf an incision was made through the corium
in the left groin, scrapings from different ulcers on the teats were
well rubbed in with the blade of the scalpel, and a portion of crust
inserted into a small pocket in the subcutaneous tissue. In the ears
and the groin there were positive results. In the large brown calf
one of two places inoculated in the right ear passed through the
following changes : On the third day there was apparent vesiculation
and commencing formation of crust. From day to day the crust
thickened, and on the eighth day the crust was at its height and
detached at its edges. By removing the scab an ulcer was exposed ;
there was slight inflammatory thickening. About the thirteenth
day the ulcer had quite healed.
Very similar appearances resulted in the ear of the smaller calf.
The result of inoculation in the groin was of a very much severer
character. In the course of two or three days the incision had
apparently commenced to heal by scabbing, but there was a surround-
ing area which was inflamed, and painful on manipulation. The
inflammatory thickening which resulted continued to increase around
the seat of inoculation, and the thickening could he felt to extend
deeply into the groin. Suppuration followed, and on firm pressure
pus welled up through the wound. The wound then showed very
little disposition to heal, and the calf began to exhibit marked
constitutional symptoms. During the second week after inoculation
the animal became very dull, and was reported by the attendant as
refusing to feed. Diarrhea supervened, and lasted for several days,
and bloody urine was passed. The calf was also noticed to cough,
and the cough gradually increased in severity. Thirty-six days after
the date of inoculation it was decided to kill the calf and examine
the condition of the viscera. The appearances which were found
at the post-mortem examination were as follows :—
The upper and middle lobes of each lung were adherent to the walls
of the chest; there was congestion, especially of the middle lobe, and
patches of recent adherent lymph. Posterior parts of the upper lobes of
both lungs were completely consolidated, and on section varied in colour
from brick-red to greyish-white. The interlobular tissue was infiltrated
with inflammatory products, which mapped out the tissue of the lung in
small indurated areas, in which the tissue was granular-looking and friable.
THE AUTHOR'S INVESTIGATION. 273
These appearances in the upper lobes were due to septic pleuro-pneumonia.
They closely resembled, and were supposed to be due to, infectious
pleuro-pneumonia. They were, however, found identical with the con-
dition observed in septic pleuro-pneumonia in calves, and the disease was
not conveyed by infection to other animals in the same stall. Scattered
through the other lobes of both lungs were white, mostly firm, nodules
raised above the level of the surface of the lung. They were surrounded
by a zone of congestion, and in some cases sections were composed of
indurated, in others of friable, lung tissue. In the posterior part of the
right upper lobe there was a recent infarct. The bronchial glands at the
roots of each lung were enlarged to two or three times their‘ natural size,
and were firm and hardonsection. The parietal surface of the pericardium
was covered with recent adherent lymph. The visceral surface of the
pericardium was normal. Along the external surface of the aorta were
chains of enlarged lymphatic glands connected by dilated lymphatic
vessels. These glands were dark red or purplish in colour, from hemor-
rhage into their substance. The heart was normal, and the endocardium
not stained. There were chains of red glands on the cesophagus similar
to those along the aorta. The appearance of the mesenteric glands was
very striking. The mesentery, along the lymphatic vessels, was dotted
with glands, varying in size from a large shot to a pea, which were deep
red or prune-coloured. In addition, there were here and there enlarged
glands without hemorrhage into their substance, and greyish in colour.
There were scattered petechiz on the spleen. The kidneys were firm
on section, and there was marked congestion in both, while it was more
pronounced in one kidney than the other. The liver was congested, the
congestion being more marked in patches.
Sections from the consolidated upper lobes showed under the micro-
scope thickening of the pleura and infiltration with round cells. The
exudation filled the alveoli, and was breaking down in some cases in the
centre. The vessels were injected, and there were hemorrhages into the
alveoli. The periphery of the lobules was infiltrated with round cells.
In sections of the kidney there was slight infiltration around glomeruli
and arterioles with round cells; the epithelium in the convoluted tubules
was granular and disintegrating ; there was hemorrhage in the straight
tubules, and engorgement of vessels. In sections of liver the inter- and
intra-lobular vessels were engorged ; there were interlobular collections
of round cells displacing the liver cells, and the interlobular connective
tissue was infiltrated with round cells; the liver cells were granular and
cloudy.
There can be no doubt from the symptoms and post-mortem
appearances that this calf had been suffering from septicemia as
the result of introducing the septic virus and crust subcutaneously
in the groin.
The two Wiltshire cows were killed, and there was nothing of
importance to note in one, but in the other an incision into the
udder revealed an enormous abscess,
18
274 INFECTIVE DISEASES.
Though the naked-eye appearances of the kidney in this case
were practically healthy, the results of examining sections of the
kidney under the microscope were extremely instructive and interest-
ing, as they showed that marked changes had taken place which
were indicative of septic complication.
The sections showed glomerulo-nephritis ; there was infiltration
of the capsule of Bowman with round cells; there was infiltration
also of the sheaths of the vessels with round cells, especially in
the cortex. The blood-vessels in the boundary zone of the medulla
were engorged, the arterioles of the glomeruli were also engorged,
and there were slight hemorrhages into the capsule. The epithelium
of the convoluted tubules was granular, opaque, and in some parts
breaking down.
Sections of the ulcers of the teats of these cows were also
carefully examined, and the appearances corresponded exactly with
the description given by Klein.
On visiting the farms it was found that there were altogether
about a hundred and sixty cows. Only a few had proved refractory,
and had not taken the disease at all. The rest had contracted the
disease in varying degrees of severity. About fifty at a time were
dry, and they escaped until they were in milk again. The milk
was drunk on the farms and in the village, and a quantity was
supplied to a large town. Most careful inquiries were instituted to
ascertain the existence of scarlatina among consumers of the milk,
So far the research was completely analogous to the Hendon
investigation ; but, in spite of the contamination of the milk, no
cases of scarlatina were found either on the farms or in the village,
and there was no epidemic in the town in which the milk was
distributed.
The disease, in fact, was cow-pox, and in no way connected with
scarlet fever; and to assist others who may undertake a similar
inquiry the details will now be given of the author's investigation
into the nature of the outbreak in Wiltshire.
THe Disease PROVED TO BE Cow-Pox.
Locality of the Wiltshire Outbreak.—There is considerable interest
attached to the fact that the farms were situated a few miles from
Cricklade. They are close to the borders of Gloucestershire, and about
twenty-five miles from Berkeley. They are, therefore, within that
district in which in Jenner's time cow-pox was particularly prevalent.
Time of Year.—The outbreak commenced about the end of September
1886, and lasted until about the middle of December. In an outbreak
AN OUTBREAK OF COW-POX. 275
in 1885, a few miles from these farms, but on a separate estate, the
disease appeared in June and July.
Origin of the Outbreak.—The author made careful inquiries as to the
origin of the outbreak, but beyond ascertaining with certainty that the
disease appeared first at one farm, and was conveyed from this to the
other farms, all evidence was negative. The milkers were unable to
say whether it commenced in one particular cow or whether it broke
out in several simultaneously.
The only information which could be obtained, which was very
suggestive, was that the milkers were in the habit of receiving their
friends from neighbouring farms on Sundays. The friends would assist
in the milking, to get the work done as quickly as possible on these
occasions. As it was reported that the same disease had occurred
that summer on a neighbouring farm, it is quite possible that it was
introduced by one of the milkers’ friends.
Mode of Dissemination.—When the disease made its first appearance,
the bailiff, attributing it to the farm being, for some reason, unhealthy,
decided to remove the cows to other farms. The herd was therefore
divided and sent to two other farms. From these cows the disease was
communicated to healthy cows, and, as this interchange was repeated,
not only of the cows, but of the milkers, the disease was communicated
to four separate farms.
In all cases the disease was limited to the teats, and was conveyed
from the teats of a diseased cow to the teats of a healthy cow by the
hand of the milker. In no case was there any evidence of the disease
being produced in healthy cows by other means than contact.
Bulls and dry cows remained free from the disease, while the cows
in milk, numbering about a hundred and twenty, were all attacked, with
the exception of about a dozen, which proved to be entirely refractory.
These facts explain how it is that the disease has been known from
time immemorial as the “ cow-pox.” We never hear of cattle-pox or
bull-pox. We have not, in other words, to deal with an infectious
disease like cattle-plague or pleuro-pneumonia, but with a disease which
is communicated solely by contact.
The disease was only observed in the cows in milk, and was limited
to the parts which come in contact with the hand of the milker. The
virus was mechanically transferred from diseased to healthy cows, being
communicated to all, or nearly all, the animals in the same shed,
whether the milker had vesicles on his hand or not.
Character of the Eruption on the Cow.—In a recent case which was
carefully examined the teats were visibly inflamed, partly red and partly
livid in colour. On each teat there were vesicles, some broken, and
others which appeared to be just forming. In other cases there was
nothing more than the remains of broken and dried vesicles, and more
or less characteristic crusts on the teats.
On visiting a byre at the time that the cows were brought in to
be milked, it was a striking sight to look along the line and see one
animal after another affected with the eruption ; and thus one character
276 INFECTIVE DISEASES.
of the disease was clearly shown—the tendency to spread through a
whole herd.
On examining the eruption carefully, the degree of severity was
found to differ very much in different animals. In a few cases the
condition was most distressing, both to the cow and to the observer. In
such cases the teats were encrusted with huge, dark brown or black
crusts, which, when handled in milking, were broken and detached,
exposing a bleeding, suppurating, ulcerated base. Such ulcers varied
in size from a shilling to a florin, and in form were circular, ovoid, or
irregular. Weeks afterwards, when the animals had recovered, the
site of these ulcers was marked by irregular scars.
All the milkers agreed as to the general characters of the malady,
laying particular stress on the teats being red, swollen, and painful when
handled. Vesicles would then appear on the teats—two, three, four, or
more on each teat. They were soon broken in milking, and irritated into
sores, which became covered with thick crusts. From four to six weeks
elapsed before they had entirely healed. Other more observant milkers
insisted that before the teats were red and swollen, spots or pimples first
appeared which came to a head. This head increased if it was not broken,
which might be the case if it was situated between the bases of the teats,
until it formed a greyish vesicle of the size of a threepenny-piece or
even larger.
General Symptoms in the Cow.—As to the general condition of the cows
nothing abnormal was observed. They appeared in the best of health,
and in only one particular was any difference from their condition in
health stated to exist. This was, that in the majority of the cases there
could be no doubt that the milk had diminished. This might escape
notice by inexperienced milkers in any particular animal, but the total
amount of milk supplied by the herd was considerably below the average.
History of the Eruption communicated to the Milkers.—The most striking
characteristic of this outbreak was the communicability of the disease
to the milkers. A milker, with vesicles which presented typically the
characters of casual cow-pox, was taken to London and kept under
observation. The various cases will be described in the order in which
they first presented themselves, their history being given as much as
possible in their own words.
CasE I.—J.R., milker, informed the author that he was the first to catch
the eruption from the cows. He stated that it came as a hard, painful spot,
which formed “ matter” and then a “ big scab.” He had been inoculated
about seven weeks previously. He pointed to the scar which remained
on his right hand. This scar presented the characters of an irregular
cicatrix, indicating considerable loss of substance. He stated that he had
also two places on his back, where he supposes he had inoculated himself
by scratching. He had continued milking ever since, but had had no
fresh places. :
Case II.—W. H., milker. He stated that he was inoculated from the
cows about the same time as J. R. They were the two milkers of
the herd in which the cow-pox first made its appearance. The eruption
AN OUTBREAK OF COW-POX. 277
appeared in one place on each hand. He pointed to two irregular scars
as the remains of the eruption.
Case ITI.—J. L., milker, stated that he also caught the disease from
the cows. On his right hand a spot appeared which formed a blister,
then discharged matter and produced a bad sore. Lumps formed at the
bend of his elbow and in his armpit. He lost his appetite, felt very
poorly, and was obliged to leave off work for two or three days.
Case IV.—W. K., « labourer on the farm, was put on as a milker to
take the place of one of the others with bad hands. After his fifth or
sixth milking—that is to say, about three days after first milking the cows,
—pimples appeared on his hands, which became blistered and then ran on
to bad sores. He pointed to three irregular scars on the first and third
fingers and palm of the right hand. Lumps appeared in his elbow and in
his armpit, but he did not feel very poorly in consequence.
Case V.—J. F., milker, stated that about a month ago he noticed
spots which appeared on both hands. His fingers swelled and were pain-
ful. He said it came first like a pimple, and felt bard. Then it ‘ weeped
out” water in four or five days. There were red marks creeping up to
his arm. There was a sort of throbbing pain, and he could not sleep at
night. On the right hand there was a scar, but on the left hand there
was an ulcer about the size of a shilling covered with a thick black crust.
The crust was partially detached, and exposed a granulating ulcer. It
was in this stage the exact counterpart of the ulcers on the cow’s teats.
Case VI.—W. H., junior, milker, stated that he had both hands bad
about a month previously: first on the index finger of the left hand,
and then on the right hand on his knuckle and between the first and
second fingers. He said that it came up like a hard pimple, and the
finger became swollen and red. After a few days it “weeped out”
water, and then matter came away. Both his arms were swollen, but
his left arm was the worst. About a fortnight after, he noticed kernels
in his armpits, which were painful and kept him awake at night. His
arms became worse, he could not raise them, and he had to give up
milking. He also had had a “bad place” on the lower lip. On examina-
tion, I found that the axillary glands were still enlarged and tender.
He volunteered the statement that the places were just like the sore
teats.
Case VII.—J. H., the bailiff’s son, also milked the cows. He had
a sore on the upper lid of his right eye and on his left hand. In both
cases he had been previously scratched by a cat, and the scratches were
inoculated from the cow’s teats. Theright hand also had been inoculated.
The eruption broke out a fortnight previously. His hands were swollen,
red, and hot. He felt very poorly and went to bed. Little spots like
white blisters appeared on the back of his right hand. His mother
remarked that they “rose up exactly as in vaccination.” Thick dark
brown scabs formed. He was very ill for two or three days, but did not
send for a doctor. He had painful lumps at the bend of his arm and in
the armpit. He gave up milking, and had not taken to it since.
On examining him, the thick crusts on his right hand were identical
278 INFECTIVE DISEASES.
with the stage of scabbing in vaccinia. The scabs fell off in about three
weeks to a month, and left permanent, depressed scars.
Case VIII.—W. P., milker. This case was pointed out on the occasion
of another visit, and is the only one in which the eruption was seen in
its earlier stages.
The history of this boy is as follows. He had taken the place of one
of the other milkers who had vesicles on his fingers, and had been obliged
to give up milking. After the seventh time of milking he noticed a
small pimple on his right cheek. This became larger and vesicular.
On examination it presented a depressed vesicle with a small central
yellowish crust and a tumid margin, the whole being surrounded by a
well-marked areola and considerable surrounding induration. On raising
the central incrustation a crater-like excavation was seen, in which
lymph welled up and trickled down the boy’s cheek. On the following
day the crust had re-formed, and was studded with coagulated lymph.
The areola became more marked, and on pricking the margin of the
vesicle, the exuding contents were slightly turbid.
From this day the surrounding infiltration increased enormously,
the whole cheek was inflamed, and the eyelids so cedematous that the
eye was almost closed. There was enlargement of the neighbouring
lymphatic glands. The crust which had re-formed thickened day by day.
It retained the character of central depression, and was situated on a
reddened, raised, and indurated base (Plate VII.).
From this date the surrounding induration gradually diminished.
The crust changed in colour from dark brown to black, and finally
fell off, leaving an irregular, depressed scar. This scar, when seen several
months afterwards, was found to be a permanent disfigurement. The
eruption appeared on the fourth day after exposure to infection, and
allowing two days for incubation, the vesicle was at its height on the
seventh or eighth day, and a typical tamarind-stone crust fell off on
the twenty-first day after infection, leaving a depressed, irregular
cicatrix.
A vesicle also formed on the thumb of the left hand. Two days
after the pimple appeared on his cheek, the lad said that he first noticed
a pimple on his thumb, and this, on examination, presented a greyish
flattened vesicle, about the size of a sixpence. Later, its vesicular
character was much more marked, and a little central crust had com-
menced to form. The margins became very tumid, giving it a marked
appearance of central depression. The vesicle was punctured at its
margin with a clean needle, and from the beads of lymph which exuded
a number of capillary tubes were filled.
Two days afterwards suppuration had commenced, the vesicle con-
tained a turbid fluid, and the areola was well marked. Later, the crust
had assumed a peculiar slate-coloured hue, and, on pressing it, pus
welled up through a central fissure. The areola had increased, and
there was considerable inflammatory thickening. The lymphatic glands
in the armpit were enlarged and painful. Though there was deep
ulceration, which left a permanent scar, the ulceration did not assume
DESCRIPTION OF PLATE VII.
Casual Cow-pox.
Fic. 1—Case of W. P——, a milker, infected from the teats of a cow with
natural cow-pox. There was a large depressed vesicle with a small
central crust and a tumid margin, the whole being surrounded by a
well-marked areola and considerable surrounding induration.
Fig. 2.—The same case a week later, showing a reddish-brown crust on a
reddened elevated and indurated base.
Plate VIL.
CASMAT YCOW PO.
oo Vincent Brocka, Day & Son, Li
AN OUTBREAK OF COW-POX. 279
quite so severe a character as in some of the other milkers. Possibly
this may be accounted for to some extent by the fact that the pock was
covered with a simple dressing instead of being subjected to the irritation
and injury incidental to working on the farm.
Revaccination of the Milkers.—There were in all eight milkers, varying
in age from seventeen to fifty-five, who had vesicles on their hands
from milking the cows. Seven had been vaccinated in infancy, but not
since ; one had been revaccinated on entering the navy at fifteen. They
were all revaccinated by a public vaccinator after complete recovery
from the casual cow-pox (that is to say, from three to four months
afterwards), and were all completely protected. On the other hand,
two of the three milkers who had escaped infection from the casual
cow-pox were also vaccinated, with the result in one of typical
revaccination, in the other of very considerable local irritation.
Retro-vaccination of Calves.—The result of retro-vaccinating calves with
the humanised lymph was strictly in accordance with the experience of
Ceely, who has pointed out that in retro-vaccination from the milker’s
hands the results are doubtful, and depend greatly on the animals selected.
“Those of a light colour and with thin skins were generally preferred,
but often without avail, scarcely one-half of the operations succeeding.”
“Vaccine lymph, in passing from the cow to man, undergoes « change
which renders it less acceptable and less energetic on being returned to
many individuals of the class producing it ; some refuse it altogether.”
Two cases out of four succeeded, and an eruption was produced with
all the typical characters of vaccinia, but running rather a rapid course,
and the protection passing off after a few weeks, while the result
obtained in calves inoculated with pus or scrapers from ulcers was in
accordance with what is well known to occur if pus instead of lymph
is taken for carrying on calf to calf vaccination.
That the cow-pox in Wiltshire was identical with the so-called
Hendon cow-disease there can be little room for doubt, for in both
cases we find that—
1. The disease spread through a whole herd of milch cows.
2. The disease was characterised by the appearance of vesicles,
which were broken by the hand of the milker, and irritated into
deep ulcerations.
3. The disease was conveyed from one cow to another by the
hand of the milker.
4, The vesicular eruption was communicable to the hand of
the milker.
5. The disease was not fatal, and in cows which were killed and
examined the post-mortem appearances could not be distinguished
from accidental complications.
6. The naked-eye appearances and the duration of the ulcers of
the teats were the same.
280 INFECTIVE DISEASES.
7. Sections of the ulcers showed under the microscope identical
appearances of a cellular character, and the purulent discharge of
the ulcers contained pyogenic cocci.
8, The results produced by inoculation of calves with the septic
virus were identical.
If we examine the chain of argument which has been brought
forward to maintain the existence of cow-scarlatina at Hendon, we
find that it was urged :—
1. That the Hendon cow disease was a disease in which the
post-mortem appearances resembled scarlatina,
2. That this disease was associated with a streptococcus, which
produced, by inoculation in calves, a disease with post-mortem
appearances similar to those of the Hendon cows.
3. That a streptococcus regarded as identical with the one
above mentioned was found in certain cases of scarlatina in man,
which when inoculated in calves produced post-mortem appearances
similar to the post-mortem appearances in the original Hendon cows
and in certain cases of scarlatina in man.
But the microscopical appearances of the kidney of a Wiltshire
cow were identical with those which were regarded as indicating
scarlatina in a Hendon cow; and, indeed, the statements as to the
post-mortem appearances in the Hendon cows, when studied, not
only do not necessarily indicate scarlatina, but they cannot even be
considered of primary importance, or as throwing much light on the
question of scarlatina at all. The description of the naked-eye
appearances in both cows only suggests coincident pleurisy or
pleurisy with pneumonia. The microscopical appearances in both
were suggestive of septic complication.
A careful examination of the post-mortem appearances of calves
inoculated with scraping of an ulcer of a Hendon cow, or with
cultivations of the streptococcus from certain cases of scarlatina,
brings to light much more striking changes. These appearances,
however, cannot be regarded as indicative of scarlatina. They are
in reality the post-mortem appearances of septic poisoning, and
oceur commonly in many diseases. This is clearly shown by com-
paring the post-mortem appearances in the calf which was killed
while suffering from septicemia as the result of inoculation from
the ulcers of a Wiltshire cow. These visceral changes are not to
be distinguished from the post-mortem appearances described in
the calves inoculated by Klein. Consequently, that the strepto-
AN OUTBREAK OF COW-POX. 281
coccus found in certain cases of scarlet fever should produce on
inoculation in calves certain post-mortem appearances which are
found in many diseases, and should fail to produce fever, ulceration
of the tonsils, or scarlatinal rash, or any condition in the least
resembling, clinically, the disease in man, and yet that the result
should be regarded as scarlatina in the calf, is a conclusion quite
untenable.
It is true that visceral lesions similar in character were produced
in calves whether inoculated with scrapings or with streptococci from
ulcers of the Hendon cows or with streptococci from certain cases of
scarlet fever. In both cases the streptococcus is pathogenic, and
inceulation of Streptococcus pyogenes or the inoculation of septic
virus, is liable to produce septicemia. These facts constitute a mass
of evidence which justifies the conviction that the pathological data
which appeared to support the theory that the vesicular disease of
the teats of cows at Hendon was scarlatina in the cow, admit of an
entirely different interpretation, and there can be no longer any
doubt that the milk was not infected by the cows but with the virus
of scarlet fever from some human source which Mr. Power failed to
discover.
All the other evidence reported to the Board of Agriculture pointed
to the same conclusion. The disease at Hendon was admittedly
introduced from Derbyshire; and from Professor Axe’s report it
appears that only a part of the herd was sold to the farmer at
Hendon; other cows with the same eruption were transferred to
other dairy farms, and the disease communicated to healthy cows as
at Hendon, but in no instance did scarlet fever occur among the
consumers of the milk. At the farm of the brother of the dealer the
disease was communicated to three of the milkers, and the eruption
diagnosed by Dr. Bates as vaccinia.
All this evidence must be regarded as conclusive. The con-
tamination of the milk at Hendon with scarlet fever must neces-
sarily have been a mere coincidence; and the conclusion that
the milk could not possibly have become infected from any
human source is untenable. Professor Axe even ascertained
that scarlet fever existed at Hendon during several months of
1885, and that the dwellings where cases occurred stood within six
hundred yards of the cowsheds which contained the incriminated
cows, and that out of fourteen men on the farm six lived in
a district where cases occurred. Professor Axe has also stated
that the father and brother of a girl with scarlet fever, visited the
dairy during her illness. Whether any of those engaged on the
282 INFECTIVE DISEASES.
farm suffered from latent scarlet fever does not appear to have
been ascertained.
There is, it is true, no evidence to show that any one daily carried
infection to the milk, but the exact path of infection is not always
easy to trace; and because it was not actually traced it was hardly
reasonable to assume that the possibility of contamination from a
human source could be altogether eliminated.
In attempting to communicate scarlet fever to cows Professor
M‘Fadyean confirmed the negative results which had been experi-
enced in some earlier experiments by Klein In 1882 Klein
inoculated and fed cows and yearling heifers with diseased products
from human patients, using desquamated cuticle and the discharges
from the throat; but the experiments all failed. M‘Fadyean’s
failures were still more marked. Cows and calves were inoculated’
with blood from scarlet fever patients, and they were made to drink
water thickened with desquamated cuticle, but all the experiments
proved unsuccessful.
The author believes that the outbreak at Hendon was one
of cow-pox, which was prevalent in this country in 1886. The
outbreak in Wiltshire could not be distinguished bacteriologically
or clinically or in its micropathology, from the disease at Hendon,
and the Wiltshire outbreak proved on investigation to be true cow-
pox. This conclusion was questioned at the time, as cow-pox was
generally believed to be extinct in England; but that view is
entirely fallacious, and the author’s conclusions have since been fully
confirmed by independent observers, whose work will be referred to
in another chapter (p. 321).
Stamping-out System.—The Notification Act of 1890 may he
voluntarily adopted in sanitary districts, but it would be a great
advantage if notification were carried out uniformly all over the
country, Prompt information may lead to detecting the origin of
cases of scarlet fever, and isolation and disinfection will assist in pre-
venting its spread. Epidemics have occurred on a large scale owing
to scarlet fever existing among those engaged in dairy work, and
the precaution not being taken of stopping the milk supplied to the
consumers. Scarlet fever cannot be so readily controlled as small-
pox, for it may be spread by mild cases before the nature of the
disease is suspected, and small-pox cannot be conveyed in milk.
MEASLES. 283
MBEASLEs.
Measles is a contagious disease peculiar to man. It lasts for
one or two weeks, and produces fever, catarrh of the respiratory
mucous membrane, anda characteristicrash. It is highly contagious,
especially before the nature of the disease is revealed; there is
consequently great difficulty in preventing its spread in schools
and households. The contagium appears to be given off from the
body, principally if not entirely, by the breath. One attack is pro-
tective against future attacks. “The whole population of « country
may acquire a certain degree of immunity. Measles introduced into
countries where it was previously unknown assumes a most malig-
nant form. There are no characteristic post-mortem appearances.
Bacteria in Measles.—Micrococci have been found in the
blood, catarrhal exudation, and skin, by Keating, Babes, and others,
but they are accidental epiphytes of no importance, or associated
with secondary complications, as in scarlet fever.
Canon and Pielicke have found in the blood small bacilli varying
in form. They do not grow on nutrient agar or blood serum, but
cultures were obtained by pricking the finger of a patient suffering
from measles, and allowing the blood to drop into sterilised broth.
After a few days the broth became cloudy, and later, a floceulent
deposit formed. The bacilli were also obtained from the nasal and
conjunctival secretions. The nature of the contagium of measles
is unknown.
Stamping-out System.— Measles is not easily controlled by the
stamping-out system ; it is, in fact, extremely difficult, almost impos-
sible, to prevent its spread, as it is especially infectious during the
period of incubation. Notification, isolation, and disinfection assist in
controlling an epidemic, but the value of the system does not apply
to thesame extent in measles as in other infectious diseases.
CHAPTER XX.
SMALL-POX.—CATTLE PLAGUE.
SMALL-POX.
SMALL-POx is an infectious and inoculable disease of man, charac-
terised by sudden and severe fever, followed in forty-eight hours by a
characteristic papular eruption which gradually becomes vesicular
and then pustular. The virus is contained in the vesicles, and in a
concentrated form in mature pustules. It also passes into the air
from the breath and skin. Infection may occur from the dead body,
and clothes and bedding may retain the contagium for months.
One attack, as a rule, gives immunity against future attacks.
Small-pox is undoubtedly a disease foreign to this country. Its
home is in the East. Some of the old writers held that it spread to
Europe from Alexandria about the year 640 a.p., following in the
wake of the Arab conquests in Egypt, Palestine, Persia, along the
Asiatic coast, through Lycia, Gallicia, along the coast of Africa,
and across the Mediterranean to Spain ; others maintained that it
was not introduced until the end of the eleventh or beginning of the
twelfth century, by the returning Crusaders. At any rate, small-
pox was imported from the East, and probably from Egypt. Hero-
dotus, who visited Egypt, leads us to infer that epidemics were
unknown there during the rule of the Pharaohs; but Egypt
undoubtedly became a hotbed of pestilence during the Mohammedan
occupation. Prosper Alpinus imagined that both the plague and the
small-pox were concocted in the putrid waters of the Nile, but he
would probably have been more correct if he had suggested that
they arose from the insanitary condition of the Arab conquerors
and their filthy camp followers, who did their best to destroy all
that remained of that magnificent civilisation which had existed in
the days of the ancient Egyptians.
We do not know the exact period at which small-pox was first
imported into England, and the records of the disease are very
meagre until the sixteenth century.
284
SMALL-POX. 285
In 1593 Simon Kellwaye appended to his work on the Plague
a short treatise on the small-pox. ‘ Oftentimes,” he wrote, “those
that are infected with the plague are in the end of the disease
sometimes troubled with the small pocks or measels, as also by good
observation it hath been seen that theyare fore-runners or warnings of
the plague to come.” According to Kellwaye the disease arose from
the “excrements of all the foul humours in our bodies, which striving
with the purest doth cause a supernatural heat and ebullition of our
blood, always beginning with a feaver in the most part.”
Small-pox steadily increased in the seventeenth century until it was
a formidable scourge, for no advantage was taken of all the experi-
ence which had been gained in dealing with the plague. No public
measures were adopted to cope with the disease, and the people came
to regard the new pestilence as a visitation which was unavoidable.
Early in the eighteenth century, small-pox inoculation was introduced,
and this was superseded in the nineteenth century by vaccination.
Examination of small-pox cases after death does not reveal any
characteristic lesions in the internal organs, but sections of small-
pox vesicles show an important structure. A vesicle is formed by
the exudation raising up the outer layer of epidermis, and the chief
feature is the formation of a vacuolated structure in which, especially
in the later stages, bacteria are found in abundance.
Bacteria in Small-pox.—Cohn and Weigert found cocci in
variolous lymph. Hlava found Streptococcus pyogenes in the pustules,
and Garré streptococci in the internal organs in a case of variola
hemorrhagica. In a fatal case of variola complicated with pemphigus
Garré found a streptococcus in the pemphigus vesicles. Klein
and Copeman have found a small bacillus which they regard as
characteristic, but its biological characters are unknown, as it will
not grow on any nutrient media. The bacteria commonly found in
variolous pus are the usual pyogenic organisms. The nature of
the contagium of small-pox is unknown.
Protective Inoculation.—Experience had taught that a person
was not, as a rule, attacked with small-pox a second time; but when
and how the method of artificially inducing a mild form of the
disease was discovered, or when this preventive treatment was first
employed, is unknown. Avicenna of Bokhara was credited with the
discovery, and it was supposed that the practice was carried by
Tartar and Chinese traders to Surat, Bengal, and China, and by
the Mahommedan pilgrims to Mecca. In Constantinople it was
supposed by some to have been introduced from the Morea by
an old woman, and by others by the women of Circassia. The
286 INFECTIVE DISEASES.
Circassian women fastened three needles together, and pricked the
skin over the pit of the stomach and heart, the navel, the right
wrist, and the left ankle. The variolous matter was applied to the
bleeding points, and the eruption came out in five or six days. In
Constantinople scarifications were made on the forehead, wrists,
and legs, and carefully selected virus applied to the incisions. The
needle used was a three-edged surgeon’s needle, or the operation was
performed with a lancet. The virus was obtained by pricking the
vesicles, and pressing out the matter into a clean glass vessel. The
Armenians preferred to be inoculated in both thighs. In Barbary
a slight wound was made between the thumb and forefinger, and
the virus obtained from a mild form of small-pox. In Hindustan
the operation was performed at certain seasons of the year, and a
preparatory regimen enforced. The inoculators were very careful
in the selection of the virus, as they had learnt its varying intensity,
and they were credited with being able to control the amount of
the eruption. They preferred to inoculate the outside of the arm,
midway between the wrist and the elbow in males, and between the
elbow and the shoulder in females. The skin over the part to be
inoculated was first well rubbed with a piece of cloth; then, with
slight touches of a small instrument, little wounds were made over
an area which might be covered by a small coin, and sufficient to
cause just an appearance of blood. A pledget of cotton-wool
charged with the variolous matter, and moistened with water, was
applied to the wound. This virus was obtained from inoculated
pustules of the preceding year.
In China the contents of the variolous pustules were dried and
kept for several years. If the virus was to be used from fresh
pustules the “acrimony” of the matter was corrected by steaming.
The dried powder was made into a paste, which was wrapped up in
cotton-wool and introduced into the nostril.
The Greeks were more cautious in their procedure, and were
said to inoculate tens of thousands without an accident. They
operated only upon those in perfect health, punctures were made
with needles, and the virus was used in the crude state, freshly
obtained from the “kindly” pustules of a young child. They were
particularly careful in the choice of the ‘“ ferment.”
Dr. Perrot Williams, in 1722, wrote that the practice of com-
municating small-pox had long been employed in South Wales. The
oldest inhabitants said that it had been a common practice with
them “time out of mind,” but Lady Mary Wortley Montagu was
responsible for the general adoption of small-pox inoculation in
SMALL-POX. 287
England by persuading physicians in London to employ it. Lady
Mary had her child inoculated in Turkey. An old Greek woman inocu-
lated one arm, and Mr. Maitland, surgeon to the Embassy, the other.
The disease ensued in due course with an eruption of a hundred
pustules. This was the first time that the Byzantine method of
inoculation was performed upon an English subject. In 1721 Dr.
Harris delivered a lecture before the College of Physicians, and
described the successful inoculation of four children of the French
consul at Aleppo, by means of a thread imbued with variolous pus.
A daughter of Lady Mary was inoculated in England by Maitland in
1721, and subsequently a number of criminals were inoculated by
him. Incisions were made through the cutis, and pledgets which had
been steeped in variolous pus from ripe pustules, were applied to the
wound. This was known as Maitland’s or the reformed operation,
but it was soon modified, as troublesome ulcers resulted. Shortly
afterwards Maitland encountered another obstacle. The child of a
Mr. Batt was inoculated, had plenty of pustules, and soon recovered,
but six of Mr. Batt’s domestic servants, ‘who all in turn were
wont to hug this child while under this operation, and whilst the
pustules were out, never suspecting them to be infectious, were all
seized at once with the right natural small-pox of several and very
different kinds.”
Dr. Jurin in 1729 reverted to the Eastern method, and recom-
mended virus from a mild case of small-pox, but the virus was still
taken from perfectly maturated pustules, and the operation continued
to be followed by bad results. In order to diminish the risks, Burgess
in 1766 advocated certain improvements. An incision about an inch
long was made on each arm through the cuticle, but not so deep as
to wound the c2llular tissue. A variolous thread was laid along the
whole length of the wound and fixed with plaster. Ulcerations and
other accidents continued to take place, and a new epoch in the
history of inoculation was the introduction of the Suttonian method,
in 1764-6.
It was said that Mr. Sutton, with his assistants, inoculated
one hundred thousand persons. The method was kept secret at first,
but the essential points were all discovered and published by Dr.
Dimsdale, Dimsdale recommended a very slight puncture with a
lancet wet with variolous matter. Subsequently, Sutton published
an account of his method, and the result of his operation may be
given in his own words.
“The lancet being charged with the smallest perceivable quantity
(and the smaller the better) of unripe, crude, or watery matter, immediately
288 INFECTIVE DISEASES.
introduce it by puncture, obliquely, between the scarf and true skin,
barely sufficient to draw blood, and not deeper than the sixteenth part of
an inch. Neither patting, nor daubing of the matter, in or over the
punctured part, is at all necessary to its efficacy. This practice, indeed,
is rather prejudicial than otherwise, as it may affect the form of the
incision, and thus be apt to confound our judgment upon it.
“ Indications of the Incision—In the incipient state of variolous
increase in the incision, a small florid spot appears on the part of access,
resembling a flea-bite in size; and on passing the finger lightly over it a
hardness is felt not larger than a small pin’s head. This florid appearance
and hardness denote that the variolous principle is effectually imbibed,
and their indications point no farther, unless the progress to vesication
be very slow, in which case an uncomfortable number of pustules may be
expected to follow. The florid spot in most instances of inoculation is
somewhat larger, or more extended on the second, than on the third day
after the insertion.
“About the fourth day from inoculation, should the incision begin
to vesicate, an itching sensation will be complained of on the place of
insertion—the occurrence of which symptom is the first indication of a
favourable event, yet not of sufficient importance to justify any present
relaxation in the preparatory proceedings.
“ The vesication of the incision in most instances will begin to be visible
on the fourth or fifth day after the insertion of the matter ; the sooner
it becomes so, the more favourable may be expected to be the event. The
extent or diameter of the vesication at this stage does not usually exceed
that of a large pin’s head, and it has invariably a dint or small depression.”
Adams obtained still more striking results by inoculating with
variolous lymph from pear]-pox, a mild variety of small-pox. Starting
with lymph obtained from this benign form of small-pox, and
selecting the cases, and carrying on arm to arm variolation, the
results obtained were practically identical with the phenomena
obtained by inoculation of the arm with cow-pox lymph. Similar
results were obtained by Guillou, but more rapidly. In 1827 there
was an epidemic of variola, and Guillou, having no vaccine virus,
took variolous lymph from a girl fifteen years of age on the fifth day
of the eruption. The case was one of varioloid or mild small-pox, attri-
buted to previous vaccination. The variolous lymph was inserted in
ten places on the arm of a healthy infant still at the breast. This
inoculation produced ten beautiful “vaccine” vesicles, from which, on
the ninth day, forty-two infants were inoculated under the eyes of
two of the local authorities. These furnished virus for the inoculation
of one hundred, who were inoculated in the presence of magistrates
and many medical men. This experiment was repeated with success.
Variolous lymph was taken from two lads at school, and in ten
SMALL-POX, 289
cases produced appearances with a perfect similarity to ordinary
vaccination, :
Thiele produced a benign vesicle in the following manner.
Variolous lymph was diluted with warm cow’s milk, and inoculated
like ordinary vaccine lymph. Large vesicles resulted. There were
_ febrile symptoms from the third to the fourth day, and a secondary
onset of fever much more pronounced between the eleventh and
fourteenth days. The areola was strongly marked, and not con-
fined to the inoculated place, which was occasionally surrounded
by minute secondary vesicles. After watching through ten removes,
the vesicles finally assumed the characters of an ordinary vaccination
with cow-pox lymph. As soon as the secondary fever ceased to
occur inoculation was practised from arm to arm without diluting
the lymph with cow’s milk. The lymph was designated lacto-varioline,
and the result was variolation in its mildest form. The result of
variolating the cow will be discussed in another chapter.
Small-pox inoculation, or variolation, protected the individual
when genuine small-pox was produced, and ,endangered the com-
munity. Persons inoculated became centres of infection, and con-
veyed the disease to others. Haygarth, although in favour of
inoculation, strongly condemned its use without precautions to
prevent the spread of the disease. ‘The most serious and solid
objection,” he wrote, ‘“‘ that has been advanced against inoculation
is deduced from a comparison of the Bills of Mortality for a series
of years in various places. They show that a larger proportion
of inhabitants have died of the small-pox in towns where it is prac-
tised than in the same before it was known, or in others where it is
prohibited.”
Even Dr. Dimsdale, an ardent inoculator, admitted that more
lives were lost in London than before inoculation commenced, and the
practice was more detrimental than beneficial to society; and he
added: ‘The disease by general inoculation throughout London
spreads by visitors, strangers, servants, washerwomen, doctors, and
inoculators, by means of hackney coaches in which the sick are sent
out to take the air, or by sound persons approaching them in the
streets. The poor in London are miserably lodged ; their habitations
are in close alleys, courts, lanes, and old dirty houses ; they are often
in want of necessaries, even of bedding. The fathers and mothers
are employed constantly in laborious occupation abroad, and cannot
attend the inoculated sick.” In 1798 Jenner, who had practised
small-pox inoculation, proposed the use of a benign non-infectious
lymph obtained from a disease of the cow or horse as a substitute
19
290 INFECTIVE DISEASES,
for variolous lymph, and in 1840 small-pox inoculation was prohibited
by Act of Parliament.
Stamping-out System.—The disappointing and dangerous
results of small-pox inoculation led to a widespread demand for
some new method for dealing with small-pox. This induced Haygarth
to turn his attention to the subject, and towards the end of the
eighteenth century to bring before the medical profession and the
public a plan for stamping out the disease. Haygarth, who was a
close observer and an able physician, studied the question of the
communicability of the disease from one person to another, and its
conveyance by infected clothing and other means, and ultimately
drew up rules and regulations for its prevention, the importance of
which we are only now beginning to fully acknowledge. Haygarth’s
essential doctrine was “ that mankind was not necessarily subject to
the small-pox, and that it was always caught by infection from a
patient or the poisonous matter,” and might be avoided by observing
his Rules of Prevention.
These rules comprised a regular system of notification and isolation.
Inspectors were to be provided to report cases of small-pox, and people
were to be rewarded for carrying out the instructions. Several examples
were given of the results at Chester, where the plan was adopted.
Haygarth met with considerable encouragement from some of the
leaders of the profession. Dr. Fothergill wrote to him in 1778, saying,
“T have mentioned the intention of freeing this country from the
small-pox to divers of the faculty, and shall continue to do so as it falls
in my way. The proposal is variously received, but in exact proportion
to their humanity.”
In 1793 Haygarth made considerable addition to his rules, and urged
that legislation should follow to make them compulsory. Provision was
to be made to reward the poor for observing the rules, and public thanks
to the wealthy for their support were to be published in the parish church
and newspapers. Transgression of the rules was to be punished by a fine of
from £10 to £50, one half to go to the informer and the other half to the
fund which supplied the expense of rewards to the poor, and all details were
to be supplied to the press. It was further suggested that Great Britain
should be divided into districts, including a certain number of parishes
or townships, and that to each of them a surgeon or apothecary should
be appointed as inspector to see that the regulations were exactly
observed. In addition, there were to be directors of inspectors, superin-
tended by a commission of Physicians in London and in Edinburgh. All
salaries were to be paid by the county rates, and the rewards for observing
the rules of prevention were to be guaranteed out of the parish funds.
On the requisition of the director and inspector of a circuit, power
was to be given to two or more justices of the peace to appoint a separate
house for the reception of patients with the small-pox. In conclusion,
SMALL-POX, 291
Haygarth maintained that the plague had been completely exterminated
from this country, for above a century, by civil regulations, and that
there seemed to be little doubt that the small-pox was propagated on
principles similar to the plague, and that it also might certainly be
exterminated from this island.
Haygarth’s teachings had a profound influence upon both the profession
and the educated public, but his system of compulsory notification was
never carried out, for no legislation followed to enforce his recommenda-
tions. This is a matter deeply to be regretted, for towards the end of the
eighteenth century small-pox was declining in London, general sanitation
was making rapid advances, small-pox inoculation, which created fresh
centres of infection, was falling into disfavour, small-pox hospitals were
built, which served to limit centres of infection, and the profession and
the public were influenced by the teaching of Haygarth with regard to
the various ways of avoiding the spread of the disease.
It only required the compulsory adoption of Haygarth’s system
uniformly all over the country to have kept the disease in control,
if not to have entirely extirpated it from Great Britain. That a
similar conviction existed at the time is evidenced by an article which
appeared in 1779 inthe Medical and Chirurgical Review, in which
the following statements were made :—
‘Plans for the extirpation of the small-pox have been suggested. . . «
To do this, however, the exertions of the physician are incompetent unless
they be aided by the powerful hand of Governments, but this has hitherto
been withheld. The grand means, however, of extirpating this destructive
malady is an early and strict separation of the infected from those that
are sound.”
Small-pox in the present century has been largely controlled by
legislation, especially in recent years, by the Public Health Acts for
England and Wales, for Scotland, and for Ireland; the Epidemic and
other Diseases Prevention Act ; the Public Health Amendment Act ;
the Labouring Classes’ Dwellings Acts ; the Housing of the Working
Classes Act ; the Public Health (Ships) Act; the Local Government
Board Act-—and various orders and memoranda of the Local Govern-
ment Board ; the Infectious Diseases Notification Act ; the Infectious
Diseases Prevention Act ; and the Public Health (London) Act.
By the Public Health Act of 1875 England was divided into
Urban and Rural Sanitary Districts, and powers were given to
‘enforce regulations of the Local Government Board for guarding
against the spreading of infectious diseases; to provide medical aid
and accommodation for infected persons, to promote cleansing,
ventilation, and disinfection, to provide hospitals, to provide for
292 INFECTIVE DISEASES.
destruction or disinfection of infected bedding, clothing, and other
articles, and to appoint Medical Officers of Health.
As to the value of notification and isolation in cities such as
Tiondon we have the evidence of the Metropolitan Asylums Board.
In their Report for 1889 we read in reference to the diminution ot
small-pox : “These very satisfactory results confirm the view taken
by the Committee two years ago to the effect that the rapid and
systematic removal from crowded districts of infected persons, each
of whom might have become a centre of contagion, is an important
factor in stamping out small-pox from the metropolitan population.
The notification of cases will also greatly facilitate the action of the
managers in this direction.”
More recently there has been a most striking confirmation of
these statements. An outbreak of small-pox occurred in Maryle-
bone, and by the energy of the officials of the Board this outbreak
was suppressed in a few days by means of notification and
immediate isolation.
The Isolation Hospitals Act of 1893 gives power to County
Councils to provide, or cause to be provided, an isolation hospital
in any district within their county. An application to a County
Council for the establishment of an isolation hospital may be made
by any one or more of the authorities defined as local authorities
having jurisdiction in the county or any part of the county.
Further, the County Council may direct an inquiry to be made by
two medical officers of health in the county as to the necessity of an
isolation hospital being established for the use of the inhabitants
of any particular district in the county, and in the event of such
medical officers reporting that such a hospital ought to be
established for the use of the inhabitants of a district, may take the
same proceedings in all respects for the establishment of such
hospital, as if a petition had been presented by a local authority for
the establishment of an isolation hospital for the district named in
the report of such medical officers of health.
Lastly, the Local Government Act of 1894 provides for the
formation of District Councils; and the powers, duties, and
habilities are principally those which were conferred by the Public
Health Act of 1875,
In the opinion of the author the Government of this country should
enter into friendly negotiations with the Governments of other countries,
so that there, might be concerted action to prevent an avoidable
disease like small-pox. Much good might result from the formation of
a permanent International Board of Health. If civilisation is not yet
CATTLE PLAGUE. 293
sufficiently advanced to admit of a system of international notification,
our Consular authorities should be instructed to give immediate notifica
tion of the existence of small-pox in other countries, and every measure
should be enforced to diminish the possibilities of importation. The
duties of a Central Health Office, presided over by a Minister of Healtb,
should include the collection of information as to the existence of small-
pox in other countries, and details should be published in the Annual
Reports of the Department. Regulations, for example, for dealing with
the importation of rags from small-pox stricken places should be enforced,
as in the case of cholera ; and if, in spite of these precautions, isolated
cases occurred in this country, they should be dealt with promptly.
Notification should be enforced uniformly all over the country, and
there is not the slightest reason why the authorities and the public should
not immediately receive information of the existence of small-pox, whilst
to procure immediate isolation we have only to imitate the excellent
ambulance system of the Metropolitan Asylums Board. To procure
prompt notification there must be no loophole for evading the Act, and
there should be a heavy penalty for failure to notify not only small-pox,
but any case which may reasonably be supposed to be one of small-pox.
The police should be required to report any case of small-pox in
common lodging-houses or shelters ; they should have power to require
any tramp suffering from small-pox, or from any disease which may
reasonably be supposed to be small-pox, to be examined by the medical
officer of the Union, and kept under observation, or transferred at once
to the isolation hospital ; and inmates of the workhouse should be daily
inspected, and no case allowed to leave when there is the least suspicion
of small-pox infection.
Objections no doubt will be raised to this proposal, but the frequency
with which small-pox is spread by tramps fully justifies these measures.
All these measures should be carried out as 4 matter of routine, and
without the semblance of panic.
Isolation should be uniformly enforced all over the country, and
vaccination should be relegated to the position of a voluntary auxiliary
measure, which should never be allowed to take the place of sanitary
regulations to stamp out the disease.
CarrLe PLaGuE.
Cattle plague is a highly contagious disease of bovines producing
high fever, and characterised by an eruption with a resemblance to
human small-pox. The disease is transmissible to other ruminants,
and is inoculable in man. One attack gives immunity against
future attacks. Cattle plague and small-pox are not intercom-
municable, and are specifically distinct diseases, but the resemblance
between them was recognised from early times. Ramazzini published
an account of the cattle pest in Italy in 1711, and described the
pustules which broke out over the body as similar to those of variola in
294 INFECTIVE DISEASES.
kind and appearance. Dr, Layard, in 1780, described this disease of
horned cattle as an eruptive fever of the variolous kind, with the
appearance and stages of small-pox. This resemblance was endorsed
by Murchison, one of the Commissioners appointed in 1866 to
inquire into the origin and nature of cattle plague.
Murchison pointed out that in both diseases the eruption con-
sisted of pustules and scabs, and that in both it extended from
the skin to the interior of the mouth and nostrils; in both, the
pustules and scabs were preceded or accompanied by patches of
roseola ; in both, they were occasionally interspersed with petechie ;
and in both, they sometimes left behind pitted scars and discolora-
tions on the cutis. The other prominent symptoms of rinderpest
were also those of small-pox—viz., pyrexia, lumbar pain, salivation,
and running from the nostrils, alvine flux, albuminuria, hematuria,
and “the typhoid state.” The anatomical lesions of the internal
organs in rinderpest and unmodified small-pox were identical—viz.,
congestion or inflammation of the mucous membranes of the air
passages and digestive canal, patches of ecchymosis and even
gangrene of the stomach and other mucous surfaces, and darkly
coloured blood. In both rinderpest and small-pox the duration
of the pyrexial stage was on an average about eight days. In
both diseases a peculiarly offensive odour was exhaled from the
body before and after death. The two diseases resembled one
another in their extreme contagiousness, and in the facility, with
which the poison was transmitted by fomites. Both diseases were
easily propagated by inoculation, and in both cases the inoculated
disease was milder and less fatal than that resulting from infection.
In both diseases there was a period of incubation, which is shorter
when the poison has been introduced by inoculation than when it
has been received by infection.
Ceely described the result of an accidental inoculation of cattle-
plague virus in the human subject. A vesicle was produced which
so closely corresponded with the result of inoculated cow-pox that
Ceely inclined to the belief that cattle plague was a malignant form
of cow-pox. The following is the account of this case as reported
by Ceely. Mr, Hancock, a veterinary inspector at Uxbridge, was
engaged in superintending the autopsy of a bullock recently dead of
cattle plague. His assistant, who was performing the operation,
while occupied in removing the skin from the scrotum, accidentally
punctured the back of Mr. Hancock’s hand with the point of the
knife. The puncture being slight was disregarded at the time, but
was washed as soon ag practicable, and thonght of no more. Five
.
CATTLE PLAGUE. 295
days afterwards, a small, slightly elevated, hard pimple was felt
and seen on the site of the puncture. This gradually advanced
till the ninth day of the puncture, the fourth from papulation,
when the enlargement became distinctly vesicular, At that time
there were but slight constitutional symptoms. On the next day,
the tenth from the receipt of the puncture, the fifth from papulation,
and the second from vesiculation, Mr. Hancock consulted Mr. Rayner,
of Uxbridge, who, on seeing the hand, inquired if the patient had
been handling the udder of a cow, as he theught he could recognise
a cow-pox vesicle of the ninth day. The vesicle was distended
with thin lymph, its margin elevated and slightly brown, its centre
depressed and brownish, and the whole surrounded with a large
bright red areola. There was then considerable tumefaction extend-
ing from the knuckles to above the wrist. The absorbent vessels
were considerably inflamed, and, like the axillary glands, were tender
and painful; the pulse, naturally slow, was accelerated; there was
much pain in the back and limbs, severe distracting headache, etc. ;
all of which symptoms continued to increase during the two following
days. At the end of that time the diffused areola had extended as
far asthe elbow. Fifteen days after the puncture, and ten days after
papulation, the local inflammation and constitutional symptoms had
partially subsided. The vesicle contained a rather turbid brownish
fluid, and there were present all the indications of a declining vaccine
vesicle.
Murchison also saw and described the case, and gave practically
the same account of it. He pointed out that the appearances and
the entire history were very different from the results of a poisoned
wound, but coincided with the appearances seen after vaccination.
In 1832 Macpherson, in Bengal, inoculated eleven native children
with cattle-plague crusts. There was no result in six, others
suffered from local inflammation, and in one a vesicle formed.
With lymph from this vesicle other children were inoculated. The
results in all were similar in appearance to those of vaccination.
Two children were subsequently inoculated with human variola, and
were said to be protected.
In i834 Macpherson’s example was followed by Mr. Furrell
in Assam. Furnell inoculated four children with cattle-plague
crusts without result, but his assistant succeeded with crusts taken
from the back and abdomen of the diseased cattle, and carried on
the lymph from child to child. In one case there was a general
eruption. Furnell inoculated his own child from one of the native
children : a copious eruption followed, and the child died. Furnell
296 INFECTIVE DISEASES.
after this misfortune issued a strong warning against taking the
virus from the cow. The experiments were made in the belief that
cattle plague was really small-pox in cattle, and that the virus
would protect against human variola.
Similar results were obtained by Mr. Wood at. Gowalpara
in 1838.
Bacteria in Cattle Plague.—Semner cultivated streptococci
from the blood and lymphatic glands of a sheep suffering from
cattle plague. A calf inoculated with a cultivation died in seven
days. The cocci were stated to lose their virulence by cultivation,
and the weakened cultivation to protect against the virulent disease.
The micro-organism was very probably Streptococcus pyogenes,
and the calf may have died of septic infection. There can be no
doubt that the nature of the contagium of cattle plague is unknown.
Protective Inoculation.—In the great epidemic of cattle
plague in England in 1866, owing to a belief that the analogy
between cattle plague and small-pox was closer than it really is,
vaccination with cow-pox was attempted as a preventive measure,
but was proved to be absolutely useless.
Stamping-out System.—When cattle plague was imported in
1865 into London, dairymen and stock-owners made no attempts
to prevent the extension of the disease, so that it spread rapidly
all over the country through disposal of infected cattle. The losses
were enormous, and an Order in Council was passed in July 1865,
directing dairymen and others to notify outbreaks of any contagious:
or infectious disease among the animals under their charge. <A
Veterinary Department of the State was formed, and inspectors
appointed in various parts of the country. A short Act was passed
in February 1866. A stamping-out system, consisting of compulsory
notification and the slaughter of diseased animals, was soon brought
to the notice of the public. There was violent opposition, but
nevertheless, after some delay, the system was carried out. The
number of cases of cattle plague had reached 18,000 weekly, and
on the introduction of the stamping-out system the disease rapidly
declined. The disease was again imported into Great Britain in
1872, and there were outbreaks in 1877. In each instance the
disease was promptly stamped out, and ever since that year the
disease has been kept out of this country.
CHAPTER XXT.
SHEEP-POX.—FOOT-AND-MOUTH DISEASE.
SHEEP-Pox. °
SuZEP-Pox, or variola ovina, is an acute febrile disease accompanied
by a general vesiculo-pustular eruption, highly infectious, and
capable of being propagated by inoculation or clavelisation. It is
a common disease in some parts of Europe. In France the disease
is called la clavelée, and in Italy vaccuolo. It has been introduced
on several occasions into this country, but has been effectually
stamped out. As in human small-pox, there are varieties—the
benign and the malignant, the discrete and the confluent ; and one
attack is protective against the disease in future.
It is very closely analogous to human small-pox. Vaccination
with cow-pox lymph has been employed to protect sheep from sheep-
pox, but unsuccessfully, and lymph for vaccination has been raised
from sheep-pox to protect human beings from small-pox. These
experiments were first performed in Italy.
Marchelli, in 1802, took lymph from the vesicles of sheep-pox, and
inoculated children. Sacco repeated these experiments, and found
there was no appreciable difference from the results obtained with
cow-pox lymph. Dr. Legni carried on the inoculations with ovine
virus from arm to arm for several years, and when small-pox
occurred in Pesaro, it was said that all those who were inoculated
with the sheep virus were protected.
Inoculation of children with ovine virus, direct from sheep, was
repeated by Sacco and Magnani in 1806.
Marson in England succeeded in producing on the human
subject a vesicle with the physical characters of the vaccine vesicle.
The vesicle had a bluer tinge, and subsequent inoculation of the
patient with human variola was ineffectual. Other experimenters
were unsuccessful, but their failures, as in the case of variolation of
the cow, do not invalidate the results of those who were successful.
297
298 INFECTIVE DISEASES.
Sheep-pox and cow-pox are quite distinct diseases. Sheep-pox
is highly infectious, whereas cow-pox is only conveyed by direct
inoculation, and is never infectious, and further, cow-pox inoculated
in sheep does not produce sheep-pox.
Bacteria in Sheep-pox.—Hiallier and Zurn, Klein, and others,
have found micrococci and bacteria in the lymph of the vesicles of
sheep-pox, but they are only accidental epiphytes. The nature of
the contagium is unknown.
Protective Inoculation.— Extensive experiments were carried
out in England to test the protective power of vaccination against
sheep-pox. According to Marson and Simmonds, it was very difficult
to get cow-pox to take on sheep, and when an effect was produced,
the resulting affection, even when developed to its fullest extent, was
very unlike the same disease in the human subject. In the sheep
it seldom produced anything more than a small papule, which occa-
sionally resulted in the formation of a minute vesicle, or more
commonly, a pustule, which was sometimes, although very rarely,
surrounded by a slight areola. Generally, however, neither vesica-
tion nor pustulation followed, but a small scab was produced, which
soon fell from the site of the puncture, leaving no trace behind. The
disease passed quickly and irregularly through its several stages,
and terminated by the eighth or ninth day, and not unfrequently
even before that time. Lymph was but rarely obtainable, and then
only in the smallest quantity, and this on the fifth or sixth day suc-
ceeding the vaccination. The effects were only local, and the animal’s
health was not impaired.
Sheep were found to be just as susceptible of the cow-pox virus
on subsequent repetition of the inoculation as they were in the
first instance, and hence the conclusion that cow-pox was utterly
worthless as a protective against sheep-pox. According to Depaul,
however, cow-pox takes characteristically on sheep, and sheep-pox
lymph inoculated on cows produces a result indistinguishable from
the appearances obtained with the inoculation of cow-pox lymph.
It is impossible to say whether these conflicting results depended
upon the employment in the experiments of different breeds of sheep
or different stocks of vaccine lymph.
The objection to clavelisation or ovination is that the disease
may be introduced in localities where it was previously unknown.
By ovination, as in the analogous case of variolation, fresh centres
of infection are created, whereas every precaution should be taken
to prevent the introduction of the disease.
Stamping-out System.—Sheep-pox has been imported into this
FOOT-AND-MOUTH DISEASE, 299
country on several occasions. It was introduced in 1847, and again
in 1862; in 1865 it was introduced again, and active measures of
repression were at once taken. The diseased flocks were carefully
isolated, and day by day as fresh cases occurred the diseased animals
were killed and buried. Owing to the adoption of these precaution-
ary measures, the affection did not extend beyond the flock among
which it first appeared. It was introduced again in 1866 at Long
Buckby, in Northamptonshire. In this case the disease was exter-
minated by the slaughter and burial of the whole flock, and imme-
diate application of disinfectants to the hurdles and other things with
which the sheep had been in contact. Then it was introduced
again in Cheshire, and strict isolation being enforced the infection
died out. Since 1866 we have had no outbreak of sheep-pox in
this kingdom, but foreign sheep have been landed with sheep-pox
in 1868, 1869, 1870, 1871, 1875, 1876, 1878, and 1880, but the
disease has been prevented from spreading.
The Sheep-pox Order of 1895 provides for the notification of
the disease, for disinfection and for compulsory slaughter of infected
sheep, and prohibits the movement of diseased or suspected sheep,
and the local authority may, if they think fit, order the slaughter
of suspected sheep and of sheep which have been in contact with
diseased sheep.
Foor-anp-mouTtH DisEase.
Foot-and-mouth disease is a highly contagious and infectious
febrile disease, characterised by a vesicular eruption affecting the
lips, tongue, roof of the mouth, and feet of sheep, cattle, and pigs,
and according to some observers it also attacks horses, poultry,
hares, and rabbits. Sometimes the mouth only is affected, in other
cases the principal seat of the eruption is in the feet. The vesicles
soon break and give rise to ulcers. When these occur in the mouth
they cause pain and difficulty in taking food. Extensive ulceration
may occur on the feet, causing great pain and lameness. In milch
cows it sometimes happens that the eruption occurs on the udder and
teats, and it is this manifestation of the disease which has received
so much attention from Rayer. The milk is contaminated by the
discharge of the vesicles, and is unfit for use, either as food for the
human being or for the lower animals. It induces a vesicular
eruption in the mouth, larynx, pharynx, and intestinal canal. It
acts most vigorously when administered warm to young animals,
and calves occasionally die quite suddenly after sucking cows
300 INFECTIVE DISEASES.
affected with the eruption on the teats. Fatal effects also result
when the milk is administered to young pigs.
It has been stated that no injurious consequences arise from the
consumption of the milk by human beings, but there is abundant
evidence to the contrary, and the conflicting opinions probably arise
from the fact that milk is seldom drunk direct from the cow, and
rarely in an undiluted form. Hertwig experimented upon himself
with milk freshly drawn from a cow with the eruption. He drank
a pint, and two days afterwards experienced slight fever, restless-
ness, and headache. The mouth was dry and hot, and there was
tingling in the skin of the hands and fingers. These symptoms
continued for seven days after taking the milk. On the ninth day
vesicles had formed on the tongue, principally on the edges, and on
the mucous membrane of the cheeks and lips (the largest being
about the size of a lentil). They were yellowish-white in colour, and
contained a whitish turbid liquid, which flowed when the vesicles
were pricked with a needle. At the same time a number of vesicles
developed on the hands and fingers; and most of them at the
time of their first appearance were the size of a millet seed. They
were firm to the touch, yellowish-white, and occasioned a slight
tingling. The vesicles of the mouth increased in size and eventually
broke, and the epithelium detached itself completely from the affected
parts, leaving dark red spots, which disappeared gradually. The
slight fever present during the first days ceased after the appearance
of the eruption; but from this time, until the disappearance of
the red spots, Hertwig felt a continual burning pain in the mouth,
and speaking and deglutition caused considerable uneasiness. On
the lips the vesicles dried up, and were covered with thin brownish
crusts, which fell off ten days after the appearance of the first
vesicles, The vesicles which developed on the hands ran a slower
course. From the tenth to the thirteenth day they filled with a
liquid, like turbid lymph. They were large and confluent, and
finally broke and dried up.
Bacteria in Foot-and-mouth Disease.—Klein in 1885 isolated
from the vesicles a streptococcus which in its microscopical and its
cultural characters on gelatine, agar and blood serum resembled
Streptococcus pyogenes. Minute differences in the size of the
colonies and in their rate of growth, and in the character of the
chains, were ,observed on making comparative cultures with
Streptococcus pyogenes from a human source, but no comparison
was made with Streptococcus pyogenes from acute suppuration in
cattle.. Baumgarten regarded this micro-organism as Streptococcus
FOOT-AND-MOUTH DISEASE. 301
pyogenes, and not as the contagium of the disease. The author has
pointed out the variation which exists in the size of the chains and
of the colonies, and the difference which is found in the rate of
growth of cultures of Streptococcus pyogenes, and these variations are
especially marked in Streptococcus pyogenes bovis. Klein believes
that the administrations of broth cultures produced the disease in
sheep, but the results were very probably due to accidental infection.
It is well known how very readily foot-and-mouth disease is spread.
The appearance of a case in a flock of sheep or a herd of cattle will
be almost certain to be followed by all or nearly all of the other
animals being infected with great rapidity. The virus clings to the
clothes of shepherds and others who have been in ‘contact with
infected sheep, and may be readily conveyed to healthy animals by
those who have been visiting infected premises.
Schottelius described chains composed of rounded elements, some
of which resembled an ameeba or plasmodium. The chains were said
to be motile, and delicate growths were obtained in blood serum and
agar, and in broth and on potato. Inoculation in sheep and pigs
and numerous small animals gave negative results. These organisms
were described as streptocytes, to distinguish them from bacteria.
Piani and Fiorentini investigated the contents of the vesicles, and
also described corpuscular elements exhibiting amceboid movements.
They regarded these bodies as protozoa, and concluded that foot-and-
mouth disease is due to their presence.
Until a micro-organism is cultivated which will produce sheep-
pox in sheep on a farm or on premises where the disease does not
exist, and where there can be no possibility of accidental infection,
we are fully justified in concluding that the nature of the contagium
of this disease is unknown. :
Stamping-out System.——Foot-and-mouth disease was imported
into this country in 1839. It has been successfully dealt with by the
stamping-out system, which in this case is very difficult to apply
because of the very short period of incubation, and the value of
the stamping-out method very greatly depends upon the length
of the incubation period. Foot-and-mouth disease very often,
from infection to recovery, does not exceed ten days; yet according
to the reports of the Board of Agriculture, when foot-and-mouth
disease exists in a manageable state, perfect isolation and effectual
disinfection have proved equal to the complete control of the
spreading of the infection, and the final extinction of the disease.
Nothing more is necessary in any case than to close up all the
channels through which infected matter can be conveyed; but in
302 INFECTIVE DISEASES.
order that this may be done close supervision by conscientious and
responsible officers is required ; without it the case is hopeless.
The Foot-and-mouth Disease Order of 1895 enforces notification,
isolation, and disinfection, and the question of slaughter is left to
the loca] authority.
(1) A local authority may, if they think fit, cause to be slaughtered—
(a) Any cattle, sheep, or swine affected with foot-and-mouth disease
or suspected of being so affected ; and
(b) Any cattle, sheep, or swine being or having been in the same field,
shed, or other place, or in the same herd or flock or otherwise in
contact with animals affected with foot-and-mouth disease, or being
or having been, in the opinion of the local authority, in any way
exposed to the infection of foot-and-mouth disease.
CHAPTER XXII.
HORSE-POX.— COW-POX.
ConsTITUTIONAL GREASE OR HorsE-Pox.
Horsz-pox is a vesicular disease of the horse communicable from
animal to animal by inoculation, but never infectious. It is
communicable by inoculation to man, and the attenuated virus
produces phenomena indistinguishable from the results of vaccina-
tion with cow-pox lymph.
The existence of this disease of the horse had long been known
to farmers and farriers, but Jenner was the first to draw attention
to it in writing. ‘There is a disease to which the horse from his
state of domestication is frequently subject. The farriers have
termed it the grease; it is an inflammation and swelling in the heel
accompanied at its commencement with small cracks and fissures,
from which issues matter possessing properties of a very peculiar
kind.” Jenner gave several instances in which this disease was
communicated to man and to cows.
Thus, a man named Merret attended to some horses with sore
heels and also milked the cows. The cows were infected, and the
man had several sores upon his hands.
William Smith, on another farm, attended to horses with sore
heels and milked the cows also. The cows were infected, and on one
of Smith’s hands there were several ulcerated sores.
Simon Nicholls applied dressings to the sore heels of one of his
master’s horses and at the same time milked the cows, and the cows
were infected in consequence.
A mare, the property of a dairy farmer, had sore heels, and
was attended to by the men of the farm, Thomas Virgoe, William
Wherret, and William Haynes. They contracted “sores on their
hands, followed by inflamed lymphatic glands in the arms and
axille, shiverings succeeded by heat, lassitude, and general pains in
the limbs,” and the disease was also communicated to the cows.
303
304 INFECTIVE DISEASES.
But Jenner’s experience of this disease was not limited to cases
in which the eruption occurred in the heel.
He mentions a case in which—
“ An extensive inflammation of the erysipelatous kind appeared
without any cause upon the upper part of the thigh of a sucking
colt. The inflammation continued several weeks, and at length
terminated in the formation of three or four small abscesses.” Those
who dressed the colt also milked the cows on the farm, and communi-
cated the disease to them.
Subsequently, Jenner gave a more comprehensive description of
this disease.
“The skin of the horse is subject to an eruptive disease of a
vesicular character, which vesicle contains a limpid fluid, showing
itself more commonly in the heels. The legs first become edematous,
and then fissures are observed. The skin contiguous to these fissures,
when accurately examined, is seen studded with small vesicles sur-
rounded by an areola. These vesicles contain the specific fluid. It
is the ill-management of the horse in the stable that occasions the
malady to appear more frequently in the heel than in other parts.
I have detected it connected with a sore on the neck of the horse,
and on the thigh of a colt.”
Mr. Moore, of Chalford Hill, described a case in 1797, and re-
garded the disease as virulent grease. His horse was attacked with
what was supposed to be ordinary “grease.” A cow was subse-
quently infected, and the disease communicated to the servant, who
had “eruptions on his hands, face, and many other parts of the
body, the pustules appearing large, and not much unlike the small-
pox, for which he had been inoculated a year and a half before, and
had then a very heavy burden.”
In 1798, Mr. Fewster, of Thornbury, met with a case of this
equine malady, and wrote a very full account to Jenner of its
transmission to the human subject.
“William Morris, aged thirty-two, servant to Mr. Cox of
Almonsbury in this county, applied to me the 2nd of April, 1798.
He told me that four days before he found a stiffness and swelling
in both his hands, which were so painful it was with difficulty he
continued his work ; that he had been seized with pain in his head,
small of the back, and limbs, and with frequent chilly fits succeeded
by fever. On examination I found him still affected with these
symptoms, and there was great prostration of strength. Many
parts of his hands on the inside were chapped, and on the middle
joint of the thumb of the right hand there was a small phagedenic
CONSTITUTIONAL GREASE OR HORSE-POX. 305
ulcer, about the size of a large pea, discharging an ichorous fluid.
On the middle finger of the same hand there was another ulcer of a
similar kind. These sores were of a circular form, and he described
their first appearance as being somewhat like blisters arising from a
burn. He complained of excessive pain, which extended up his arm
into the axilla, On the 5th of April I again saw him, and found
him still complaining of pain in both his hands, nor were his febrile
symptoms at all relieved. The ulcers had now spréad to the size of
a seven-shilling gold coin, and another ulcer, which I had not noticed
before, appeared on the first joint of the forefinger of the left hand,
equally painful with that on the right. I ordered him to bathe his.
hands in warm bran and water, apply escharotics to the ulcers, and
wrapped his hands up in a soft cataplasm. The next day he was
much relieved, and in something more than a fortnight got well.
He lost his nails from the thumb and fingers that were ulcerated.”
Mr. Tanner, a veterinary surgeon, was the first to succeed in
experimentally transmitting horse-pox to the teats of a cow by
inoculating some of the liquid matter from the heel of a horse. From
handling the cow’s teats he became infected himself, and had two
pustules on his hand, which brought on inflammation, and made
him unwell for several days. The matter from the cow and from his
own hand proved efficacious in infecting both human subjects and
cattle.
In 1801 Dr. Loy published his experiments. A butcher had
painful sores from dressing a horse suffering from ‘ grease,’ and Dr.
Loy succeeded in transmitting the disease to the udder of a cow.
Matter was taken from the cow and inserted into the arm of a child.
Dr. Loy also inoculated a child direct from a horse suffering from
‘ grease,’ and subsequently five other children from this child.
From his experiments and observations Dr. Loy was led to
differentiate constitutional grease from the merely local affection
commonly known as the grease, and thus he explained the failure
on the part of many experimenters to transmit this disease to.
the cow.
“This fact induces me ”—he says—‘ to suspect that two kinds
of grease exist, differing from each other in the power of giving
disease to the human or brute animal ; and there is another circum-
stance which renders this supposition probable. The horses that
communicated the infection to their dressers were affected with a
general as well as a topical disease. The animals at the commence-
ment of their disease were evidently in a feverish state, from which
they were relieved as soon as the complaint appeared at their heels,
20
306 INFECTIVE DISEASES.
and an eruption upon the skin. The horse, too, from which the
infectious matter was procured for inoculation, had a considerable
indisposition, previous to the disease at his heels, which was attended,
as in the others, with an eruption over the greatest part of his
body; but those that did not communicate the disease at all, had
a local affection only. From this perhaps may be explained the
want of success attending the experiments of the gentlemen I have
mentioned.”
Experiments with horse-pox were also made about this time on
the Continent. Sacco made some observations upon this disease at
Milan. Several horses were suffering from what was called giardoni,
and Sacco’s servant was attacked on both arms, from dressing one
of his horses troubled with this disease. Several children and cows
were inoculated from the horses, but without success. In another
instance, a coachman went to the hospital with the eruption on his
hands, and the disease was successfully communicated to three out
of nine children. é
In 1803 Dr. Marcet described some experiments which had been
made at Salonica by M. La Font. The disease was known to the
farriers in Macedonia as javart. In one case, a horse was attacked
with feverish symptoms that ceased as soon as the eruption appeared.
The fore legs were much swelled and several ulcers formed. M. La
Font took some of the discharge from an ulcer and inoculated a cow.
and three children, and succeeded in transmitting the disease to two
of. the latter. ;
Vaccinogenic grease was observed in Paris in 1812, and Baron
cites the case of a coachman who, after dressing a horse with the
“ grease,” had a crop of pustules on his hands, from which the disease
was. experimentally transmitted by inoculation to two children. A
series of inoculations was started from an infant who was infected
from one of the scabs taken from the pustules on the hand of the
coachman.
In 1813 Mr. Melon, a surgeon at Lichfield, met with vaccinogenic
grease in the horse, and some of the virus was sent to Jenner, who
carried on a series of arm to arm equinations for some months. And
again in 1817, vaccinogenic grease broke out in a farm at Wansell.
The farm-servants and the cows were infected, and Jenner employed
this equine matter for a series of inoculations for eight months.
In 1817 Baron described a case of a young man who had not less
than fifty pustules on his hands and wrists from dressing a horse with
this disease, and in the following year Baron obtained some fresh
equine virus from the hands of a boy who had been infected directly
CONSTITUTIONAL GREASE OR HORSE-POX. 307
from a horse. The disease assumed a pustular form, and extended
over both arms.
In 1818 Kahlert met with this equine disease in Bohemia, and
confirmed the experiments made by Loy and Sacco. Kahlert noticed
that the joint of the foot was swollen, and moisture exuded from it,
and that the posterior part of the pastern was slightly red, swollen
and hotter than the neighbouring parts, and a clear yellowish fluid
with a peculiar odour escaped. At the slightest touch the animal
showed signs of pain; the hair was stuck together. The disease was
successfully transmitted to cows and from cows to children.
In 1860 the horses at Rieumes, near Toulouse, were attacked by °
an. epizootic malady ; in less than three weeks there were more than
one hundred cases. According to the veterinary surgeon, M. Sarrans,
the animals suffered from slight fever, rapidly followed by local
symptoms, the most marked of which were swelling of the hocks, and
an eruption of small pustules on the surface of the swollen parts,
which were, at the same time, hot and painful. After three to five
days there was a discharge from the pastern which continued for eight
to ten days, during which the inflammation gradually diminished.
The pustules dried up, and in about a fortnight the crusts with patches
of hair fell off, leaving more or less marked scars. The eruption
appeared at the same time on different parts of the body, especially
on the nostrils, lips, buttocks, and vulva. Sarrans believed that the
mares taken to the breeding establishment at Rieumes had been
infected from the ropes which had been used in tying up other affected
animals, and had become thereby infected with the virus of this
disease. One of the mares was taken by the owner, M. Corail, to
the veterinary school to be examined by M. Lafosse. About eight
days after this visit significant symptoms appeared : loss of appetite,
lameness, stiffness of both pastern joints, and a hot, painful swelling
of the left pastern joint. The hair was staring, and there were
vesicles on the skin, from which a liquid exuded having an ammoniacal
odour but less foetid than the secretion in eaux aux jambes.
M. Lafosse successfully transmitted the disease to cows, and from
cows to children and to a horse.
In 1863 the subject of vaccinogenic grease or horse-pox again
received great attention in France. A student named Amyot was
engaged in dressing a horse on which an operation had been per-
formed. The leg which had been operated on became the seat of a
very confluent eruption of horse-pox, which was followed by such
an abundant flow of serosity that at first the nature of the affection
was mistaken, and it was thought to be a complication of eaux aux
308 INFECTIVE DISEASES.
jambes. Amyot had a wound on the dorsal aspect of the first inter-
phalangeal joint of the little finger of his right hand ; in spite of
this, he continued to dress. the horse entrusted to his care. The
wound on his finger became accidentally inoculated with the virus,
which flowed in great abundance from the horse’s leg.
The wound was made on August 3rd, and the next day it was
swollen, and rather painful. On the 5th, Amyot suffered from malaise
and great weakness; on the 6th, 7th, and 8th, vesicles appeared
successively on the fingers of his left hand, and on his forehead
between the two eyebrows. On the 9th, these vesicles were fully
developed; those of the fingers consisted of very large epidermic
bulle on a bluish-red base. On opening them, a perfectly limpid
fluid escaped in such abundance that small test-tubes might have
been filled with it. The vesicle on the forehead was surrounded by
a bluish-red areola, within which, the epidermis, of a leaden-grey
hue, was raised, and had a slight central depression. The liquid
which flowed from it when it was opened, and which continued to
ooze, was also very abundant and of a deep citrine colour.
The vesicles which had developed on the dorsal side of Amyct’s
fingers were extremely painful. The incessant shooting pains, of
which they were the seat, prevented him from getting any rest for
three days. On the 10th, inflammation of the lymphatics followed ;
both arms were swollen and very painful, with red lines indicating
the course of the lymphatic vessels. The glands of the axille were
also enlarged.
The lymphatic glands behind the jaws were also swollen and pain-
ful. Amyot’s chief sufferings were occasioned by the intense local
pain caused by the vesicles on the fingers, and by the inflammation
of the lymphatic vessels and glands, which continued in this state up
to the 18th of August. It was only at the end of the month that the
vesicles were completely cicatrised.
Bouley felt very great anxiety in the presence of the grave
symptoms which accompanied the eruption. The eruption on the
forehead was especially a cause of great uneasiness, because glanders
manifests itself in a similar way.
With virus from Amyot’s vesicles the disease was transmitted to
cows and to children.
Further, this outbreak enabled exhaustive experiments to be
made, by which it was definitely established that horse-pox is never
infectious, but, like cow-pox, is transmitted solely by contact.
In 1880, M. Baillet, Director of the National Veterinary School
of Toulouse, was informed that a contagious malady had developed
CONSTITUTIONAL GREASE OR HORSE-POX. 309
in the mares, which had been served by the stallions at the breeding”
establishment at Rieumes, belonging to M. Mazéres. M. Peuch
was delegated to investigate this outbreak, and he visited for that
purpose Bérat, Rieumes, and Labastide-Clermont.
At Bérat three mares were examined. In one, there were scars
and crusts, the remains of an eruption on the lips and in the vicinity
of the vulva ; in another, there were several reddish circular ulcers
in the same region; and in a third, there were dried pustules with
blackish adherent crusts at the circumference of the vulva and
extending over the perineum. On the lower part of the left flank
a vesicle was discovered surmounted by a crust, and when the latter
was detached a sero-sanguinolent liquid oozed from the exposed
surface. M. Peuch recognised the true nature of this disease,
having several times previously had the opportunity of examining
mares with a vesicular eruption round the vulva after coition,
which eruption he had studied from its first appearance to complete
cicatrisation, and had ascertained to be horse-pox.
On proceeding to Rieumes, M. Peuch inspected eleven stallions,
six horses, and five asses. In one ass there were several vesicles,
on the right side of the penis scattered about from the base to the
glans. In another ass there was a trace of a vesicle on the penis
and a characteristic vesicle on the left nostril.
In an old bay mare there were the remains of an eruption
on the circumference of the vulva, and in an old white mare there
were not only vesicles on the vulva, but in addition vesicles on the
inner side of the lower lip. M. Peuch drew special attention to
these cases as likely to be confounded with aphthous stomatitis, but
the existence of the same eruption on other parts of the body is an
important aid in making a diagnosis of horse-pox.
At Labastide-Clermont one mare was particularly noticed.
This mare had been served on the 19th and 21st of April, and on the
occasion of the inspection, May 11th, there were the remains of an
eruption around the vulva, and lymphangitis existed in the right
posterior limb, which was engorged, hot, and painful in its whole
extent, so that the animal walked with difficulty. The proprietor
had contracted the disease in attending to his mare, and exhibited
a vesicle on the thumb of the right hand, excoriated and blackened,
but still recognisable.
Some of the crusts collected from the cases at Bérat were used
for inoculating a cow. The result was successful, and the disease
was transmitted by inoculation to a heifer and several students and
children,
310 INFECTIVE DISEASES.
M. Peuch ascertained that horse-pox caused considerable alarm
from the fact that the breeders regard this eruptive affection as
syphilitic, and this alarm consequently brings discredit upon the
breeding establishment whence the illness has spread. He was also
led to appreciate the great necessity for further study of this disease
in relation to dourine or maladie du coit.
In 1882 M. Peuch had the opportunity of investigating a case
of horse-pox in Algeria. The disease occurred in a thoroughbred
Arab. There was an eruption of vesicles, and there was also an
weer the size of a five-franc piece in the nostril. In the mouth and
on the lips there were a number of small vesicles about the size of
a pea. The sublingual glands were engorged, hot, and painful on
pressure. The coat, in patches, on the lateral aspect of the neck,
on the shoulders, the flanks, and in the hollow of the heel, was
staring, giving the appearance of small paint-brushes. On passing
the hand over these, vesicles could be detected partly dry and partly
secreting.
The disease was transmitted to cows, and from cows to about
one thousand five hundred persons.
Cases similar to the one just described, in which there is more or
less marked ulceration of the nostril or nasal septum, must be care-
fully distinguished from glanders. And again, when the sublingual
glands are affected the disease may be mistaken for strangles.
NatuRE AND AFFINITIES.
Horse-pox and human small-pox are quite distinct diseases,
and the theory that horse-pox is derived from grooms or other
attendants suffering from small-pox may be dismissed without
further comment.
Horse-pox is never infectious, but is communicated solely by
contact—either by grooms inoculating the virus with their hands,
sponges, or brushes, or by horses coming into contact with each other,
and in breeding establishments by coition. Auzias Turenne, who
wrote exhaustively on this subject, maintained that horse-pox came
into the same category of diseases as syphilis in man.
“A un point de vue, le grease pustuleux inoculé offre la plus
parfaite ressemblance avec la verole inoculée, par le produit des
accidents secondaires. Des deux cétés nous voyons, absence de
contagion par la voie de latmosphére, travail local, retentissement
lymphatique et ganglionaire, fermentation universelle de l’organisme,
eruption générale et immunité acquise contre de nouvelles atteintes.
“A un autre point de vue, la ressemblance avec la variole est
NATURE AND AFFINITIES, 311
frappante. Mais il s’en distingue énormément par l’absence de la
contagiosité atmosphérique.”
Human small-pox belongs to a different group of diseases, and
has affinities rather with small-pox of sheep and cattle plague,
diseases which are not only inoculable, but highly infectious. Human
small-pox is an infectious disease characterised by sudden and severe
fever, followed after forty-eight hours by a generalised eruption ; horse-
pox commences as a local affection, and constitutional symptoms
follow, Auzias Turenne, guided by analogy, described the general-
ised eruptions following “‘grease” or horse-pox as greasides (* convme
on dit syphilides”).
Horse-pox and human syphilis are absolutely distinct diseases ;
and there is no more ground for believing that horse-pox originates
in human syphilis than thereis for accepting the theory that it arises
from grooms suffering from small-pox. Syphilis artificially inocu-
lated on the human subject only resembles the casual or intentional
inoculation of virulent horse-pox. The stages of papulation, vesicu-
lation, ulceration, scabbing, and the formation of a permanent scar,
occur in inoculated syphilis, and if we examine Ricord’s illustrations
and study the experiments of Auzias Turenne, we cannot fail to
be struck with the remarkable similarity to the results obtained and
depicted by Jenner.
But in order to follow the argument of Auzias Turenne we
must study the natwral and casual horse-pox. And if we are not
familiar with what has been written on this subject, and if we
restrict our knowledge to the artificially cultivated horse-pox, we
shall fail to recognise the disease when we meet with it, and we shall
be liable to attribute the results of the full effect of the virus to
accidental contamination.
Another question of very great interest is the relation of horse-
pox to cow-pox. Jenner first of all propounded the theory that all
cow-pox arose from horse-pox, or as he termed it “ the grease,” and
thus cow-pox and horse-pox were manifestations of the same disease.
But it was established that cow-pox also arose quite independently
of horse-pox, and Jenner was led to distinguish between cow-pox,
a disease peculiar to the cow, and the eruptive affection transmitted
to the cow from the horse, which farmers and others, by a strange
perversion of terms, called the cow-pox. Whether the eruption of
cow-pox can be distinguished from the eruption of horse-pox com-
‘municated to the cow, and whether cow-pox and horse-pox are
identical, or only analogous, are questions which call for further
investigation.
312 INFECTIVE DISEASES.
Bacteria in Horse-pox.—Outbreaks of horse-pox have not
been investigated from the bacteriological point of view, and the
nature of the contagium is unknown.
Cow-Pox.
Cow-pox is a vesicular disease of the teats of cows. It is never
infectious, and only attacks cows in milk, the virus being transferred
from cow to cow by the hand of the milker. The disease is com-
municable to milkers, and the virus artificially inoculated produces
what is commonly known as vaccinia. In its clinical history and
epidemiology cow-pox is totally distinct from human small-pox, and
the hypothetical and entirely erroneous suggestion that the disease
arises from milkers suffering from human small-pox is responsible
for the belief which prevailed until recently, that cow-pox was an
extinct disease in this country; but, by the author’s researches, this
has been shown to be a mistake.
Cow-pox is not a rare disease, and it has never been found to
arise from a milker suffering from small-pox. As this is a matter
of great importance in discussing the etiology of the disease, the
history of outbreaks and the clinical characters of cow-pox will be
given in considerable detail.
According to Jenner, cow-pox had been known among farmers
from time immemorial. He refers to cases occurring in 1770, 1780,
1782, 1791, 1794, 1796, and 1798. In 1799 cow-pox was raging in
the dairies in London, and outbreaks were investigated by Woodville,
Pearson, and Bradley. In the same year cow-pox broke out at
Norton Nibley, in Gloucestershire. Pearson and Aikin referred to
the prevalence of cow-pox in Wilts, Somerset, Devon, Bucks, Dorset,
Norfolk, Suffolk, Leicestershire, and Staffordshire; and Barry men-
tioned its prevalence in Ireland.
From this time onwards, for a long period, natural cow-pox
received little or no attention in this country. Fresh stocks of lymph
were raised for the purposes of vaccination, but no further attention
was given to studying the disease in the cow. In 1836 Leese
described an outbreak of cow-pox, and in 1838 Estlin discovered an
outbreak in Gloucestershire. In 1838-39 cow-pox was met with
by Mr. Fox, of Cerne Abbas, and again in 1839, in Dorsetshire, by
Mr. Sweeting. Ceely frequently met with cow-pox in the Vale of
Aylesbury, and particularly refers to outbreaks in 1838, 1840, 1841,
and 1845. But after this, outbreaks of this disease in the cow were
not recorded, though several medical practitioners met with the
disease and raised fresh stocks of vaccine lymph. Thus, when
COW-POX. 313
inquiries were made in 1857, it was found that Mr. Donald
Dalrymple, of Norwich (on two occasions), Mr. Beresford, of Nar-
borough, in Leicestershire, Mr. Gorham, of Aldeburgh, Mr. Alison,
of Great Retford, Mr. Coles, of Leckhampton, Mr. Rudge, of
Leominster, and one or two others, had met with outbreaks of
cOw-pox.
In 1885 cow-pox was discovered by the author in Wiltshire. The
publication of the fact led to the recognition of the disease in the
same year in many parts of England, and cases were met with in
man in 1888 by Mr. Forty in Gloucestershire, and by Mr. Bucknill
near London in 1894.
In Italy, cow-pox was found by Sacco in the plains of Lombardy
in 1800, and by other practitioners in 1808-9. In 1812 it was
observed at Naples by Miglietta ; in 1830 in Piedmont ; and in 1832
and 1843 at Rome, by Dr. Maceroni. More recently, several out-
breaks of cow-pox have been met with in this country, and the stocks
of vaccine lymph renewed.
In France, in 1810, cow-pox was found in the department of
La Meurthe, and in 1822 at Clairvaux; at Passy, Amiens, and
Rambouillet in 1836 ; at Rouen in 1839; at St. Illide, at St. Seine,
and at Perylhac, in 1841; in 1842 at Pagnac; in 1843 at Deux
Jumeaux, where, during the previous thirty years, several fresh
stocks of lymph had been raised and circulated. The disease occurred
in a cow belonging to M. Majendie in 1844, and it was found at
Wasseloune, in the department of Bas Rhin, in 1845 ; it occurred in
three other departments in 1846; at Rheims, and in the department
of Eure et Loire, in 1852; in the arrondissement of Sancerre, and at
Beziers in 1854; and at Guyonville in 1863. It broke out on farms
in three villages near Nogent in 1864 (the disease was introduced by
newly purchased cows ; milkers were infected, and from one of these
milkers a lymph stock was established) ; it also occurred in 1864, at
Petit Quevilly, near Rouen; and in April 1866 at Beaugency; in
1881 at Eysines, near Bordeaux, and again at the same place in
1883; and in 1844 at Cérons.
In Germany, as soon as attention had been drawn to the disease,
cow-pox was frequently discovered. There were as many as thirty-
eight outbreaks reported in one year in Wurtemberg.
It is hardly necessary, after reciting these instances, to insist
that cow-pox is far from being a rare disease, as many have sup-
posed who are unacquainted with the literature of the subject and
unfamiliar with the appearances of the natural disease in the
cow.
314 INFECTIVE DISEASES.
NatuRAL AND CasuaL Cow-pox.
To appreciate the characters of the natural disease in the cow,
we must dismiss from our minds the artificial disease vaccinia, for
the ordinary results of vaccination stand in much the same relation
to the natural disease cow-pox as the benign vesicle of variolation to
natural smail-pox.
The description of cow-pox given by Jenner, in 1798, was the
first published account. The disease in the cow was described as
consisting of irregular pustules on the teats, of a palish blue colour,
surrounded by an erysipelatous inflammation, and characterised by
a tendency to degenerate into phagedenic ulcers. The animals were
indisposed and the secretion of milk lessened.
In referring to an outbreak which occurred epizodtically in
London in February 1799, Dr. Bradley gave a coloured plate of the
disease on the arm and fingers of a milker. The cow-pox, he said,
in this instance, “ appears to have been very mild, for no loss was
experienced by the farmers from the deficiency of milk, as usually
happens.”
These early descriptions were supplemented by an account of
cow-pox by Mr. Lawrence, author of A Philosophical and Practical
Treatise on Horses, and on the Moral Duties of Man toward the Brute
Creation. Lawrence's article on cow-pox not only affords evidence
that this disease was known to those who had the care of cattle
before Jenner’s paper was published, but it shows that it had also
been made the subject of practical observation and study by veteri-
narians. Lawrence concluded by saying: ‘“ Whatever may be the
fate of cow-pox inoculation, it has and will give further occasion to
a pretty large and open discussion, which is always beneficial as
having a tendency to produce discovery and promote improvement ;
and when the public ardour for the present topic shall have become
a little cool and satisfied, I hope it will be turned by enlightened
men towards, another, perhaps of nearly as great consequence—
namely, the prevention of the original malady in the animals them-
selves. Those who have witnessed it and only reflected upon the
excessive filth and nastiness which must unavoidably mix with the
milk in an infected dairy of cows, and the corrupt and unsalubrious
state of their produce in consequence, will surely join me in that
sentiment.”
Lawrence was almost a century before his time. Cow-pox was not
again brought forward in this light until 1887-88, when the author
reported the contamination of the milk at the Wiltshire farms, and
NATURAL AND CASUAL COW-POX. 315
advocated the advisability of placing this disease under the Con-
tagious Diseases (Animals) Act.
The numerous pathological details wanting in the early accounts
of cow-pox were supplied by the painstaking and laborious re-
searches of Robert Ceely. From his classical papers in the Trans-
actions of the Provincial Medical Association, we can obtain a
complete picture of the natural disease in the cow.
In Ceely’s experience in the Vale of Aylesbury, outbreaks
occurred at irregular intervals, most commonly appearing about
the beginning or end of the spring, rarely during the height of
summer. There were outbreaks at all periods from August to May
and the beginning of June, cases being met with in autumn and
the middle of winter, after a dry summer. The disease was occa-
sionally epizootic, or occurring at times at several farms at no great
distance from each other, but was more commonly sporadic or
nearly solitary. It was to be seen sometimes at several contiguous
farms; at other times at one or two farms. Many years might
elapse before it recurred at a given farm, although all the animals
might have been changed in the meantime. Cow-pox had broken
out twice in five years in a particular vicinity at two contiguous
farms, while at an adjoining dairy, in all respects similar in local
and other circumstances, it had not been known to exist for forty
years. It was sometimes introduced into a dairy by recently
purchased cows. Twice it had been known to be so introduced by
milch heifers. It was considered that the disease was peculiar to
the milch cow; it came primarily while the animal was in milk,
and it was casually propagated to others by the hands of the milkers.
Sturks, dry heifers, dry cows, and milch cows milked by other hands,
grazing in the same pastures, feeding in the same sheds, and at
contiguous stalls, remained exempt from the disease.
For many years, the “spontaneous” origin of cow-pox had
not been doubted in the Vale of Aylesbury. In all the cases that
Ceely had noticed he could never discover the probability of any
other origin.
Condition of Animal primarily affected.—There was much diffi-
culty in determining at all times, with precision, whether this
disease arose primarily in one or more individuals in the same dairy.
Most commonly, however, it appeared to be solitary. The milkers
believed they were able to point out the infecting individual. In.
two instances, there could be very little doubt on this point. In
August 1838, three cows were affected with the disease. The first
was attacked two months after calving and seven weeks after
Aa
316 INFECTIVE DISEASES.
weaning, This animal was considered in good health, but it looked
out of condition. Heat and tenderness of the teats and udder were
the first noticed signs. The other two were affected in about ten
days. In December 1838, in a large dairy, a milch cow slipped her
calf, had heat and induration of the udder and teats, with cow-pox
eruption, and subsequently leucorrhea and greatly impaired health ;
the whole dairy, consisting of forty cows, became subsequently
affected, and also some of the milkers. In another dairy, at the
same time, it first appeared in a heifer soon after weaning, and in
about ten or twelve days extended to five other heifers and one cow,
milked in the same shed, affecting the milkers. And in another
dairy thirty cows were severely affected, and also one of the milkers,
It appeared to originate in « cow two months after calving. The
only symptoms noticed were that the udder and teats were tumid,
tender, and hot just before the disease appeared.
Condition of Animals casually affected.tIn some animals, it was
less severe than in others, depending on the state and condition of
the skin of the parts affected, and the constitution and habit of the
animal. It was sometimes observed to diminish the secretion of
milk, and in most cases it commonly did actually affect the amount
artificially obtained ; with this exception, and the temporary trouble,
and accidents to the milk and the milkers, little else was observed ;
the animal continued to feed and graze apparently as well as before.
The topical effects varied very much in different individuals; the
mildness or severity being greatly influenced by temperament and
condition of the animal, and especially by the state of the teats and
udder, and the texture and vascularity of the skin of the parts
affected. Where the udder was short, compact, and hairy, and the
skin of the teats thick, smooth, tense, and entire, or scarcely at all
chapped, cracked, or fissured, the animal often escaped with a mild
affection, sometimes with only a single vesicle. But where the
udder was voluminous, flabby, pendulous, and naked, the teats
long and loose, and the skin corrugated, thin, fissured, rough, and
unequal, then the animal scarcely ever escaped a copious eruption.
Hence, in general, heifers suffered least, and cows most, from the
milkers’ manipulations.
Progress of the Disease.—Cow-pox once arising or introduced, and
the necessary precautions not being adopted in time, appeared in ten
or twelve days on many more cows in succession, so that among
twenty-five cows perhaps by the third week nearly all would be
affected ; but five or six weeks or more were required before the
teats were perfectly free from the disease.
NATURAL AND CASUAL COW-POX. 317
Propagation by the Hand of the Milker.—Ceely was able to confirm
the way in which the disease was said to spread. In December
1838, on a large dairy farm, where there were three milking-sheds,
cow-pox broke out in the home or lower shed. The cows in this
shed being troublesome, the milker from the upper shed, after
milking his own cows, came to assist in this for several days, morning
and evening, when in about a week some of his own cows began to
exhibit the disease. It appears that, having chapped hands, he
neglected washing them for three or four days at a time, and thus
conveyed the disease from one shed to another. During the progress
of the disease throu gh this shed, one of the affected cows, which had
been attacked by the others, was removed to the middle shed, where
all the animals were perfectly well. This cow, being in an advanced
stage of the disease, and of course difficult to milk and dangerous
to the milk-pail, was milked first in order by the juvenile milker for
three or four days only, when, becoming unmanageable by him, its
former milker was called in to attend exclusively to it. In less than
aw week, all the animals of this shed showed symptoms of the disease,
though in a much milder degree than it had appeared in the other
sheds, fewer manipulations having been performed by an infected
hand.
Topical Symptoms of the Natural Disease.—For these, Ceely was
almost always, in the early stage, compelled to depend on the obser-
vations and statements of the milkers. They stated that for three
or four days, without any apparent indisposition, they noticed heat
and tenderness of the teats and udder, followed by irregularity and
pimply hardness of these parts, especially about the bases of the
teats and adjoining the vicinity of the udder; these pimples on skins
not very dark are of a red colour, and generally as large as a vetch
or a pea, and quite hard, though in three or four days many of
these increase to the size of a horse-bean. Milking is generally
very painful to’ the animal; the tumours rapidly increase in size,
vesicate, and are soon broken by the hands of the milker. Milking
now becomes a troublesome and occasionally a dangerous process.
Ceely adds: “It is very seldom that any person competent to
judge of the nature of the ailment has access to the animal before
the appearance of the disease on others of the herd, when the cow
first affected presents on the teats acuminated, ovoid, or globular
vesications, some entire, others broken, not infrequently two or three
interfluent ; those broken have evidently a central depression with
marginal induration ; those entire, being punctured, diffuse a more
or less viscid amher-coloured fluid, collapse, and at once indicate the
318 INFECTIVE DISEASES.
same kind of central and marginal character. They appear of
various sizes, from that of a pin’s head, evidently of a later date,
either acuminated or depressed, to that of an almond or a filbert, or
ever larger. Dark brown, or black, solid, uniform crusts, especially
on the udder near the base of the teats, are visible; at the same time,
some much larger are observed on the teats; these, however, are
less regular in form and less perfect. Some are nearly detached,
others quite removed, exhibiting a raw surface with a slight central
slough. On the teats, the crusts are circular, oval, oblong, or
irregular; some flat, others elevated, some thin and more trans-
lucent, being obviously secondary. The appearance of the disease
in different stages, or at least the formation of a few vesicles at
different periods, seems very evident. The swollen, raw, and én-
crusted teats seem to produce uneasiness to the animal only while
subjected to the tractions of the milkers, which it would appear are
often nearly as effectual as usual.” Referring again to the character
of the vesicle, Ceely says, that “ those fortunate enough to have an
opportunity of watching the disease in its progress may observe
that, when closely examined, they present the following characters :
In animals of dark skin, at this period, the finger detects the
intumescent indurations often better than the eye, but when closely
examined the tumours present at their margins and towards their
centres a glistening metallic lustre or leaden hue; but this is not
always the case, for occasionally they exhibit a yellowish or yellowish-
white appearance.”
In describing the crusts in detail, Ceely says that “large black
solid crusts, often more than an inch or two in length, are to be
seen in different parts of these organs, some firmly adherent to a
raw elevated base, others partially detached from a raw, red, and
bleeding surface; many denuded, florid, red, ulcerated surfaces, with
small central sloughs secreting pus and exuding blood, the teats
exceedingly tender, hot, and swollen. . . . In some animals, under
some circumstances, this state continues little altered till the third
or fourth week, rendering the process of milking painful to the
animal, and difficult and dangerous to the milker.”
“Tn many, however, little uneasiness seems to exist. The parts
gradually heal; the crusts, although often partially or entirely
renewed, ultimately separate, leaving apparently but few deep
irregular cicatrices, some communicating with the tubuli lactiferi,
the greater part being regular, smoothly depressed, circular, or oval.”
Ceely illustrated his classical memoir with a series of valuable
coloured drawings. One plate is a faithful picture of the disease on
NATURAL AND CASUAL COW-POX. 319
the teats as itis ordinarily met with ; the other is a composite picture,
consisting of the disease as ordinarily observed in the cow, to which
is superadded a number of depressed vesicles as they occur in inocu-
lated cow-pox. It is, however, an improvement on a plate published
by Sacco. The latter is an elaborate drawing, representing the udder
and teats of a cow, with an eruption purporting to be the natural
cow-pox. Jenner had described a bluish tint in the vesicles in
natural cow-pox, and Sacco deliberately represents the natural disease
by a highly coloured diagrammatic illustration in which he depicts
clusters of vesicles of inoculated cow-pox, coloured blue, and with
' a silvery lustre.
Hering has given a coloured plate of the natural cow-pox. On
the teats are a number of oval and circular bullous vesicles and
crusts. More recently, Layet has pointed out the same characters
in the cow-pox discovered near Bordeaux in 1883 and 1884. The
classical characters of the inoculated disease were wanting, particu-
larly the central depression. In Wiltshire, the author could only
distinguish, on the cow’s teats, globular and broken vesicles and
thick prominent crusts and ulcers, appearances which had very
little in common with the ordinary results of vaccination.
The early accounts of the severe character of the disease will
appear by no means exaggerated to those who have had an oppor-
tunity of studying the effects on the hands of the milkers, or indeed
to those who have made themselves familiar with the descriptions
given by Jenner, in some of his cases :—
“ Joseph Merret had several sores on his hands, swelling and stiffness
in each axilla, and much indisposition for several days.
“Mrs. H. had sores upon her hands which were communicated to her
nose, which became inflamed and very much swollen.
“ Sarah Wynne had cow-pox in such a violent degree that she was
confined to her bed, and unable to do any work for ten days.
“William Rodway was so affected by the severity of the disease that
he was confined to his bed.
“ William Smith had several ulcerated sores on his hands, and the
usual constitutional symptoms, and was affected equally severely a second
and a third time.
“ William Stinchcomb had his hand very severely affected with several
corroding ulcers, and a considerable tumour in the axilla.
“ Sarah Nelmes had a large pustulous sore on the hand, and the usual
symptoms.
‘CA girl had an ulceration on the lip. from frequently holding her
finger to her mouth to cool the raging of a cow-pox sore by blowing
upon it.
320 INFECTIVE DISEASES,
“ A young woman had cow-pox to a great extent, several sores which
maturated having appeared on the hands and wrists.
“A young woman had several large suppurations from cow-pox on the
hands.”
Pearson in his investigations encountered, and was informed of,
similar experiences.
“Thomas Edinburgh was so lame from the eruption of cow-pox on
the palm of the hand as to necessitate his being for some time in hospital.
For three days he had suffered from pain in the armpits, which were
swollen and sore to the touch. He described the disease as uncommonly
painful, and of long continuance.
“A servant at a farm informed Pearson that in Wiltshire and
Gloucestershire the milkers were sometimes so ill as to lie in bed for
several days.
“Mr. Francis said that cow-pox was very apt to produce painfal sores
on the hands of milkers.
“A servant of Mr. Francis said that cow-pox affected the hands and
arms of the milkers with painful sores as large as a sixpence.
“Mr. Dolling describes the disease as ‘ a swelling under the arm, chilly
fits, etc., not different from the breeding of the small-pox. After the
usual time of sickening, namely, two or three days, there is a large ulcer,
not unlike a carbuncle, which discharges matter.’
“Dr. Pulteney described the disease as causing ‘a soreness and swell-
ing of the axillary glands, as under inoculation for the small-pox, then
chilliness and rigors and fevers, as in the small-pox. Two or three days
afterwards abscesses, not unlike carbuncles, appear generally on the hands
and arms, which ulcerate and discharge much matter.’
“Mr. Bird wrote a short account: ‘It appears with red spots on the
hands, which enlarge, become roundish, and suppurate, tumours take
place in the armpit, the pulse grows quick, the head aches, pains are felt
in the back and limbs, with sometimes vomiting and delirium.’
“ Annie Francis had pustules on her hands from milking cows. These
pustules soon became scabs, which, falling off, discovered ulcerating and
very painful sores, which were long in healing. Some milk from one of
the diseased cows, having spurted on the cheek of her sister and on the
breast of her mistress, produced on these parts of both persons pustules
and sores similar to her own on her hands,”’
In more recent times these descriptions have been confirmed.
In 1836 cow-pox was discovered at Passy, near Paris. A black
cow, in very poor condition, had cow-pox six weeks after calving.
Bousquet had no opportunity of seeing the eruption in the early
stage, but on examination he found reddish-brown crusts on the
teats, which later gave place to puckered scars. The milk-woman,
Fleury, who had had small-pox, nevertheless contracted the disease
from the cow. She had several vesico-pustules on the right hand
NATURAL AND CASUAL COW-POX. 321
and on her lips. A vesico-pustule, when opened with a lancet,
discharged like an abscess.
In a letter to Mr. Badcock, dated April 3rd, 1845, Ceely referred
to another new stock of lymph raised from a milker’s hand. He
added :-— i
“Tn the enclosed lymph I see nothing unusually severe, except
on very thin skins; although the milker’s hand exhibits now rough
ulcers, one on the hand deep enough to encase a bean.”
Recent discoveries of cow-pox in England.-—After Ceely’s cases in
1840-41, no cases of casual cow-pox on the hands of milkers were
recognised as such and recorded in this country for nearly fifty years.
In the outbreak of cow-pox discovered by the author in December
1887, in Wiltshire, the disease was communicated to nearly all the
milkers. The reader is referred to the account of this outbreak,
which has already been given in the chapter on scarlet fever (p, 274).
The author’s researches were confirmed by Mr. Forty in 1888,
and Mr. Bucknill in 1895.
In June 1888 Mr. Forty, in practice at Wotton-under-Edge,
Gloucestershire, reported to the Local Government Board, that at
a farm at Alderley, an eruptive disease on the udder and teats was
occurring amongst cows, and that the farmer’s son, and other persons
engaged as milkers, had contracted an eruption like that of the cows.
The farmer’s son had been under Mr. Forty’s care suffering from an
eruption, and circum-anal piles. Mr. Forty had watched the course of
the eruption from papules to vesicles and scabbing, and concluded
that the eruption could not be distinguished from vaccinia. Klein
visited the farm, and found a number of cows with sores on the teats
and udders. The sores were of various sizes and outline, mostly
irregular, and covered with brown or black scabs. Those on the
teats were larger and more irregular than those on the udder.
Klein was shown several milkers who had had sores on one or more
fingers; one had had a bad arm with swollen axillary glands.
The farmer had also contracted the eruption; but in these persons
only scabs were visible as the remnants of their sores.
A girl of about twenty had taken the place of an incapacitated
milker, and noticed a red pimple form on the dorsal surface of her
right thumb. Hight days afterwards there was a slightly raised
circular vesicle, with dark centre and pale periphery ; the centre of the
vesicle was slightly depressed. It was just under half an inch in
diameter ; there was peripheral redness, but no marked areola, The
girl had three good vaccination marks.
Klein experimented on calves with lymph from the vesicle and
21
322 INFECTIVE DISEASES.
crusts from the cow’s teats, with the result that from both sources
an eruption was produced, which in appearance and course was like
vaccinia. With lymph from one of the calves, a public vaccinator
inoculated a number of infants, and fine vesicles developed, indis-
tinguishable from vaccinia.
In 1894 Mr. Bucknill met with a case in a milkman. He had
been milking a cow affected with cow-pox, and on the ninth day after
exposure to infection, and the seventh day after the eruption of the
first papule, there were three pocks on the fore-arm. The pocks
were elevated, circular, and umbilicated, with a dull, creamy-white
ring at the circumference, and there was well-marked induration
and extensive areola. There were four excellent marks of primary
vaccination. The vesicles contained clear lymph, and re-inoculation
of the arm failed to take. An attempt to re-vaccinate the man
with current calf lymph produced only topical irritation.
InocuLATED Cow-Pox.
Natural or Virulent Lymph.—Severe symptoms are not limited to
milkers casually infected from the cow. Under certain conditions,
artificial inoculation of fresh virus from the cow reproduces the
disease without any mitigation. Thus, in Jenner’s cases :—
“James Phipps. The incisions assumed at their edges rather a
darker hue than in variolous inoculation, and the efflorescence around
them took on more of an erysipelatous look. They terminated in
scabs and subsequent eschars.
“Susan Phipps was inoculated from the cow by inserting matter
into a superficial scratch on December 2nd. The child’s arm now
showed a disposition to scab, and remained nearly stationary for
two or three days, when it began to run into an ulcerous state,
and then commenced a febrile indisposition, accompanied with an
increase of axillary tumour. The ulcer continued spreading near
a week, during which the child continued ill, when it increased to
a size nearly as large as a shilling. It began now to discharge pus;
granulations sprung up, and it healed.”
Jenner’s lymph was employed by Mr. Cline with similar results.
“The child sickened on the seventh day, and the fever, which
was moderate, subsided on the eleventh. ... The ulcer was not
large enough to contain a pea.”
Precisely similar experiences have since been encountered, in the
early removes of fresh stocks of virulent lymph. Bousquet in France,
in his first trials with a new lymph, in 1836, made three punctures,
but he had soon to abandon this practice, because the intensity
INOCULATED COW-POX. 323
of the inflammation was sometimes so great that it spread over the
entire arm as far as the glands of the axilla. In one case, the
vesicles were enormous, and the inflammation so violent, that baths,
poultices, fomentations, and antiphlogistic diet scarcely sufficed
to reduce it. The crusts when they fell off left ulcerations which
were very slow to undergo cicatrisation. In some cases, the vesicles
which resulted hollowed out the skin so deeply that they left regular
holes. :
In the following year Estlin, in England, started a stock of fresh
vaccine virus from the cow, and found on inoculating children that
the new lymph was extremely active.
In 52 the disease was regular,
», 1 severe erysipelas,
» 4 erythematous eruptions of a violent character,
2 highly inflamed ulcerated arms,
1 no effect after twice vaccinating,
8 result unknown ; supposed to have been favourable.
”
”
68
Cultivated or Attenuated Lymph.—When cow-pox lymph has
been mitigated by successive transmission through the human subject,
or by cultivation on the belly of the calf, with careful selection
of vesicles, it will produce effects which are as follows: About
the end of the second day after insertion, or early on the third day,
a slight papular elevation is noticeable. By the fifth or sixth day,
it has become a distinct vesicle, of a bluish-white colour, with raised
margin and central cup-like depression. By the eighth day, the
vesicle is perfect. It is circular, pearl-coloured, distended with clear
lymph, and the central depression is well marked. On the same day,
. or a little earlier, the areola begins to appear, and gradually extends
to a diameter of from one to three inches, accompanied with
induration and tumefaction of the subjacent connective tissue. After
the tenth day, the areola begins to fade, and the vesicle at the same
time begins to dry in the centre; the lymph becomes opaque and
gradually concretes, and by the fourteenth or fifteenth day, a hard
mahogany-coloured scab is formed which contracts, dries, blackens, and
falls off between the twentieth and twenty-fifth days. A circular,
depressed, foveated, and sometimes radiated scar remains behind.
By selecting characteristic vesicles on the calf or on the human
subject, and by collecting the lymph at an early stage on the fifth,
sixth, or seventh day, this artificial disease, commonly known as
324 INFECTIVE DISEASES.
vaccinia, can be kept up in this comparatively mild form. But
under certain conditions, such as a peculiarity in the subject inocu-
lated, or if lymph be taken too late, there will be, just as in variolation,
tendency to revert to the full intensity of the natural virus.
Bacteria in Vaccine Lymph.—-Cohn, Sanderson, and Godlee
described micrococci in vaccinal vesicles. Quist and Ferré in 1883
investigated the same subject. Voigt in 1885 distinguished three
species of micrococcus—a diplococcus, a large coccus, anda third form.
Bauer in the same year described the presence of bacilli and spheero-
cocci. Marotta in 1886 regarded a tetracoccus as the specific micro-
organism, and Tenhot in 1887 distinguished a dozen micrococci, two
bacilli, and two yeasts. In the sameyear Garré isolated a micrococcus
which appeared to him to be the contagium, but inoculated on a child
it neither produced local vesicles nor immunity ; while Guttmann
pointed out three micro-organisms which appeared to be rather more
constantly present than others. Pfeiffer much more fully investigated
the bacteriology of vaccine lymph, and found Saccharomyces vaccine,
which was seldom present in human lymph but constantly found in
calf lymph ; sarcinve, both in human and calf lymph, including Sarcina
lutea, Sarcina tetragonus, Sarcina aurantiaca, Sarcina muscopus ;
bacteria and bacilli were found only exceptionally in human lymph,
but frequently in calf lymph. These included a bacterium corre-
sponding with Proteus vulgaris.
Three mice were inoculated subcutaneously with a drop of the,
liquefied gelatine, but the result was negative. The ‘injection of a
considerable quantity proved fatal to guinea-pigs and rabbits, a
result which was probably due to ptomaine poisoning.
There were also several bacilli which did not liquefy gelatine ;
these were not investigated.
Staphylococcus cereus albus was found very frequently, and
Staphylococcus pyogenes aureus occasionally. Pure-cultivations of
these micrococci inoculated on the skin of calves produced a rapid
local irritation, followed by vesiculation, but without the classical
characters of the vaccine vesicle. The inoculated part was com-
pletely healed in three to five days. According to Pfeiffer they
explain the so-called false vaccine.
Micrococcus pyogenes albus was almost constantly present.
Numerous other micrococci were found, but not constantly present ;
yaccine lymph being a splendid medium for the growth of micrococci.
Pfeiffer pointed out that the effects of Staphylococcus pyogenes aureus,
albus, and citreus, and of Streptococcus pyogenes on rabbits had an
important bearing upon the practice of vaccination, and he recom-
INOCULATED COW-POX. 325
mended that calf lymph should be tested before use upon children
by inoculation of the ear of a rabbit. If after two days no erysipelas
occurs in the inoculated rabbit, the absence of streptococci may be
considered as almost proved. ‘Two or three rabbits should be inocu-
lated at the same time.
The author’s researches into the bacteriology of vaccine lymph
extended over some years. They independently confirmed and
extended the results obtained by Pfeiffer. Having on several
occasions examined vaccine lymph and vaccine pus, and failed to find
a specific bacterium, the author proceeded to make a more systematic
examination of the different species of bacteria in samples of current
vaccinelymph. Pure-cultivations were obtained by plate-cultivation,
and inoculation of the surface of nutrient agar, obliquely solidified
in test tubes. Various current stocks of lymph were used in the
investigation. Among the specimens of calf lymph, No. 1 yielded a
torula, Bacillus pyocyaneus and Bacillus subtilis; No. 2, a bacterium, a
variety of proteus, Staphylococcus pyogenes aureus, and yellow bacte-
rium ; No. 3, a bacterium, micrococcus, yellow bacterium, and torula ;
No. 4, yellow micrococcus, white micrococcus, white torula, yellow
sarcina, white diplococcus, Staphylococcus cereus albus, and a mould
fungus; No. 5, yellow sarcina, Staphylococcus pyogenes aureus, yellow
micrococcus, white bacillus, Staphylococcus pyogenes albus, large
white micrococcus, yellow bacterium, and a white micrococcus. Among
the specimens of human vaccine lymph, No. 1 contained a white
micrococcus, proteus, and Staphylococcus pyogenes aureus; No. 2, a
micrococcus, a tetracoccus, a white liquefying micrococcus, and a
yellow bacterium; No. 3, white micrococcus, yellow micrococcus,
Staphylococcus aureus and flavus, a bacterium, a white micrococcus,
abacillus resembling Bacillus subtilis, Staphylococcus pyogenes cereus
and a brown tetracoccus. The author is familiar with these different
species of bacteria, and not one of them is peculiar to vaccine lymph ;
there was no bacterium constantly present in human and calf vaccine,
and there was not one which could be regarded as the contagium.
To sum up, most of them are well known saprophytic bacteria, and
some were identical with bacteria commonly found in suppuration.
Vaccine lymph is a most suitable cultivating medium for micro-
organisms, and bacteria invariably got access to the contents of the
vaccine vesicle. There is no evidence to be obtained by the present
methods of research as to the bacterial nature of the contagium of
vaccine lymph. Copeman obtained similar results, and thus con-
firmed the author’s conclusions.
Klein and Copeman have also observed minute bacilli in calf-
326 INFECTIVE DISEASES.
lymph and in variolouslymph. Numerous attempts to cultivate them
in nutrient media and in the living animal failed entirely, and the
identity of the bacilli could not be determined. Pfeiffer, Guarnieri,
Monti, Ruffer, and Plimmer have drawn attention to structures in
lymph, which they believe to be of the nature of parasitic protozoa.
These bodies have been studied, more especially in the tissues. They
are four times the size of ordinary micrococci, and are found in the
clear vacuole in the protoplasm of epithelial cells. Whether they
are really parasites or altered anatomical elements has not been
determined. No other conclusion can be drawn from all these
observations, except that the nature of the contagium of cow-pox
is unknown.
ORIGIN OF Cow-Pox.
Jenner’s original theory was that cow-pox was derived from
“‘orease,” but subsequently he distinguished between cow-pox, a
disease peculiar to the cow, and “grease,” a disease transmitted to
the cow from the horse, and the mistake of confounding these two
diseases was attributed to farmers and farriers. Thus he wrote :—
“ From the similarity of symptoms, both constitutional and local,
between the cow-pox and the disease received from morbid matter
generated by a horse, the common people in this neighbourhood
when infected with this disease, tirongt a strange perversion a
terms, frequently called it the cow-pox.”
Jenner’s theory of the origin of cow-pox has been discouraged ; so
also has the view of its being a ‘‘ spontaneous” disease in the cow,
though Ceely, after many years of research in the Vale of Aylesbury,
could never discover the probability of any other origin. Both
opinions have given way to the theory that cow-pox is small-pox
transmitted to the cow—an opinion advocated by Baron, and
supported by an erroneous interpretation of Ceely’s and Badcock’s
variolation experiments. Thus the cow-pox and grease of farmers
and farriers no longer attracted attention in this country, and as
natural cow-small-pox has never been discovered, cow-pox has been
credited with being extinct.
For a full discussion of this subject the reader is veferred to the
work by the author on the History and Pathology of Vaccination,
but the variolation experiments alluded to will be briefly mentioned.
In 1801 Gassner inoculated eleven cows with small-pox lymph,
and succeeded in one in producing phenomena indistinguishable from
the results of ordinary vaccination with cow-pox, and children were
inoculated from the cow.
ORIGIN OF COW-POX. 327
In 1828 Dr. McMichael reported that several physicians in
Egypt had obtained similar results, and children were successfully
“ vaccinated.”
In 1836 Dr. Martin, in America, inoculated the cow’s udder with
variolous lymph, and by inoculating children produced an epidemic
of small-pox with fatal cases. In 1839 Reiter of Munich, after fifty
unsuccessful attempts, succeeded in producing a vesicle, and a child
inoculated from the vesicle contracted small-pox.
In 1839 Dr. Thiele, after a number of unsuccessful attempts. to
inoculate cows with variolous virus, succeeded in producing a vesicle
with the physical characters of the vaccine vesicle, and from it a
stock of lymph was raised from which over three thousand persons
were inoculated. Thiele’s method was to inoculate the udder with
lymph, and to select for the purpose young cows which had recently
calved and had delicate skins. In England Ceely succeeded by
inoculating the vulva of a heifer. One of the punctures developed
into an enormous vesicle, which was undoubtedly variolous. His
assistant punctured his hand with the lancet which had been used ~
to open the vesicle, and febrile symptoms appeared, followed by an
eruption on the face, neck, trunk, and limbs, at first papular, then
vesicular, and finally pustular. The lymph was used in children,
and “vaccine” vesicles were produced. One child suffered from
vomiting delirium, and extensive roseola, but there was no eruption
in any other case.
In 1840 Badcock of Brighton inoculated a cow successfully, and
later succeeded in variolating thirty-seven out of two hundred cows
upon which he experimented.
In 1847 variolation of the cow was successfully performed at
Berlin, but the virus produced variola, and one of the children
inoculated died of confluent small-pox.
In 1864 Chauveau inoculated seventeen animals with virulent
small-pox lymph. Very small papules resulted, and the virus from
the papules produced variola in a child, which was infectious to others.
Klein in this country until recently was uniformly unsuccessful.
Voigt, Fischer, King, Eternod, Haccius, Hime and Simpson, have
all succeeded in inoculating cows and producing variola-vaccine.
The results of these experiments have been very generally misin-
terpreted, and claimed by some as conclusive evidence of the identity
of cow-pox and small-pox. Instead of the vesicle being regarded as
the most attenuated form of variola, the experimenters are said to
have ‘succeeded in producing cow-pow.
It is quite true that they produced phenomena indistinguishable
328 INFECTIVE DISEASES.
from the phenomena of an ordinary vaccination, but that does not
mean that they produced the disease cow-pox. The vesicle which
followed the inoculation, whether papular or vesicular, was small-pom.
Ceely, Badcock, Voigt, and others, succeeded in ingrafting the cow
with small-pox, and when suitable lymph and suitable subjects were
employed, the virus was so attenuated that a benign vesicle resulted.
Similar results were obtained by Sutton and Dimsdale, and identical
results by Adams, Guillou, and Thiele, by inoculating the human
subject with variolous lymph without first ingrafting the disease on
the cow.
Vaccination with variola-vaccine is simply a modification of the
Suttonian system of small-pox inoculation, only in. the first remove
the cow is substituted for the human subject. All those who were
inoculated with Ceely’s, Badcock’s, or Simpson’s variola-vaccine,
were not in the usual meaning of the word vaccinated; they were
not inoculated with cow-pox but they were variolated, and in such
an extremely attenuated form that the persons so variolated do
not convey the infection. By judicious selection it is thus possible
to obtain a strain of lymph from variola which, by direct inoculation
of the human subject or by first inoculating a cow, is deprived of
infectious properties, and produces on the arm the physical characters
of an ordinary vaccine vesicle. This has been regarded as a proof
of the identity of small-pox and cow-pox, but it is not so. Variola,
and cow-pox are not the only diseases caused by a virus which can
be attenuated until only a vesicle is produced with the characters of
an ordinary vaccine vesicle. The results which have been obtained
with the virus of cattle plague and of sheep-pox and horse-pox have
been given in previous chapters; and no one would urge on this
account that human small-pox, cattle plague, cow-pox, sheep-pox,
and horse-pox are all manifestations of the same disease. Cow-pox
has never been converted into human small-pox, and, in their clinical
history and epidemiology, natural cow-pox and human small-pox
are so different, that the comparative pathologist is no more pre-
pared to admit their identity than he is prepared to admit the
identity of cow-pox and sheep-pox, or small-pox and cattle plague.
Protective Inoculation.—Whether vaccination of all heifers
on a farm would protect them from cow-pox when they came into
milk is not known, the duration of the immunity in calves afforded
by vaccination having not been determined. Calves undoubtedly
have an immunity after vaccination, lasting for some weeks.
In 1896 Béclére, Chambon, and Menard experimented upon
the immunising power of the serum of vaccinated calves. They
COW-POX AND SMALL-POX. 329
concluded from experiments on animals and children that the serum
of a vaccinated calf from ten to fifty days after vaccination will give
immunity against inoculated cow-pox.' They further stated that,
whereas the immunity given by vaccination in the ordinary way
is not complete until the eighth day, the immunity obtained by
injection of the immunising serum is immediate. The serum has
also been credited with therapeutic properties and has, it is said,
proved efficacious in cases of small-pox.
Jenner believed that cow-pox did not protect against itself but
protected against small-pox, and for a century this has been a subject
of much controversy. The reader is referred to the Reports and
conclusions of the Royal Vaccination Commission.
Stamping-out System.—It would undoubtedly be an advan-
tage if cow-pox were scheduled under the Contagious Diseases Animals
Act. Cow-keepers and dairy-men, being anxious that their trade
should not be interfered with, very commonly conceal the existence
of the disease, and perhaps nothing is known about it, unless a milker
infected from the cows seeks for medical advice. The contamination
of the milk with lymph, pus, crusts, and sometimes blood, renders
it unwholesome, and therefore precautions ought to be taken to
prevent its occurrence. If the infected cows in a herd are the last
to be milked, and the milker washes his hands after the milking, the
disease will not spread.
CHAPTER XXIII.
DIPHTHERIA.
DiPHTHERIA is a specific infectious disease, especially of children,
characterised most commonly by inflammation, and infiltration with
lymph cells and fibrine, of the mucous membrane of the fauces,
pharynx, larynx and trachea, followed by necrosis of the mucous
membrane and the formation of a greyish-white false membrane,
the diphtheritic membrane. In some cases a diphtheritic membrane
forms in the stomach, intestine, the urinary organs and in wounds.
After the separation of the membrane an ulcer remains, which may
gradually heal. In the superficial part of the diphtheritic membrane
there are masses of bacteria including cocci, streptococci, and bacilli.
The diphtheria bacilli are not found in the blood or in the
internal organs. There is no doubt of the fact that diphtheria
is a disease which can be communicated from one person to
another; but the question of its origin is still a vexed one. There
is a close association with insanitary conditions and decaying
animal and vegetable refuse, and dampness. Localities with damp
houses, defective drainage, and a cold exposure, are favourable
to the development of diphtheria; but that does not necessarily:
indicate that these conditions can originate it. On the other
hand, assuming the disease to be due to a living contagium,
these insanitary conditions would afford a suitable environment |
predisposing to the development, and facilitating the spread, of the
disease. Scarlet fever and measles predispose to diphtheria; and
defective sanitary conditions, causing sore throat, may indirectly
act as a predisposing cause. A great many cases have been quoted
to illustrate the possibility of the conveyance of diphtheria by milk,
and the theory which best harmonises with all these observations i is.
the existence of a specific bacillus, which may be readily transferred
from the throat of the diseased to the healthy ; which finds also in
milk a suitable soil for its growth, and by its agency may be trans-
mitted to the consumer. Such a bacillus was discovered by Léffler,
330
and may be easily
obtained from the
throat of diphtheritic
patients in the fol-
lowing manner :—
Culture Outfit.—
Steel rods like or-
dinary knitting
needles, about six
inches in length, are
beaten out or rough-
ened at one end, and
a pledget of wool is
twisted round so as
to form a _ swab.
These swabs are
placed in clean test-
tubes, which are then
plugged with cotton-
wool. The test-tubes
and swabs are steri-
lised by heating in
the hot air steriliser
for an hour at 150° C.
The so-called culture
outfit consists of
a small box con-
taining a test-tube of
blood serum and a
swab. They can be
always kept ready for
use, and after use
should be conveyed
by hand for further
examination. The
danger of trans-
mitting virulent
diphtheritic material
by post is obvious.
When the examina-
tion of the tube has
been completed, the
DIPHTHERIA. 331
|
A
B
Fic. 126.—Free Surrace or Dipyrueritic Larynx
x 350 (Hamitton).—A, Deposit of diphtheria bacillus
on surface of false membrane; B, false membrane ;
C, mucosa; J, lymph-cells and false membrane
surrounded by meshes of fibrine ; ¢, surface of mucosa
deprived of its epithelium ; /,v, lymph-cells containing
shed epithelium.
332 INFECTIVE DISEASES.
culture outfit and its contents should be disinfected or destroyed.
To inoculate the tubes the patient, if it is possible, should be turned
to the light, the mouth well opened, the tongue depressed, and the
swab, without touching the teeth or the tongue, should be passed
straight to the tonsils or pharynx, and especially to the membranous
exudate. The swab is carefully and quickly withdrawn, and at
once very gently rubbed over the surface of the blood serum. The
culture outfit is then sent to the laboratory with full particulars,
and the tubes are placed in the incubator at 37° C., and can be
examined after twelve hours. If the throat has been disinfected
Fic. 127.—Bacititus or DIPHTHERIA; FROM A CULTIVATION ON BLOOD
Serum, x 1000 (FRANKEL and Prerrrer).
before examination, this must be taken into account, as the failure
to find bacilli would not then necessarily indicate a wrong diagnosis.
In all undoubted cases of diphtheria, growths will be obtained either
in the form of a pure-culture of the bacillus, or far more commonly
there will alsolbe colonies of various bacteria, especially Streptococcus
pyogenes.
Bacillus of Diphtheria.—Rods, straight or slightly curved,
‘3 to ‘8 mw in breadth, and 1:5 to 65 mw in length. They occur
singly, in pairs, sometimes in chains, and sometimes as short
leptothrix forms. Jn some cultures very irregular forms are
observed, the bacilli being swollen at one or both ends or thicker
in the middle portion, or the bacillus may contain oval or spherical
DESCRIPTION OF PLATE VIII.
Bacillus diphtheriz and Bacillus typhosus.
Fig. 1.—Cover-glass preparation from a pure-cultivation of Bacillus diph-
theriz on blood serum; obtained from the throat in a typical case of
diphtheria. Stained with gentian-violet. x 1200.
Fig. 2.—Cover-glass preparation from a pure-cultivation of Bacillus typhosus
on nutrient-agar; from the spleen in a case of typhoid fever Stained
with gentian-violet. x 1200. :
Plate VIII.
S DIPHTHERIA
U
Fig I. BACILL
Fig 2. BAC LES yeeros Us
ea, Day & Son, Lith
Vincent Broot
| EM Crackshank: fecit.
DIPHTHERIA. 333
elements, They differ greatly in size and shape, often in the same
cultures, and still more in cultures obtained from different sources.
Spore formation is unknown, In unstained preparations there are
highly refractive elements which correspond with the deeply stained
parts of the bacillus. They stain readily with the ordinary aniline
dyes. At certain stages of their growth they stain irregularly, the
protoplasm of the rod being broken up into irregular segments.
The bacillus is non-motile, and does not liquefy gelatine; it grows
at 20° C., but much more readily at higher temperatures. Colonies
in gelatine plate-cultivations are
yellowish-brown, and opaque,
granular, and circular, but with
more or less irregular margin.
In plate-cultivations on agar
and on glycerine agar the same
description applies.
On the surface of gelatine
the appearances depend greatly
on the method of: inoculation.
The growth may occur in the
form of a whitish film, but if a
sub-culture has been prepared
from broth the growth is often
composed of a number of iso-
lated white colonies (Fig. 128, a).
On blood serum, after twelve
hours the colonies appear in the
form of little elevated greyish-
white or pearl-grey dots, which
a
coalesce, forming a film if the
Fie. 128.—Purk-cuLtures or BactLius
i A DIPHTHERIA ON GELATINE: 4, isolated
of 1 per cent. alkaline glycerine colonies ; 6, filmy growth.
serum is moist. On the surface
agar, the appearances are found
to vary, and this medium is not so suitable for the cultivation of the
bacillus. In slightly alkaline broth, with or without the addition of
1 per cent. grape-sugar, the culture is cloudy, or a fine granular
deposit occurs along the sides and bottom of the tube, while the
broth remains clear.
On potato the growth is almost invisible, in the form of a dry,
thin glaze. Irregular forms are very numerous on microscopical
examination, whilst the rods are thicker than usual (Welch and
Abbott). In milk the organisms grow readily.
334 INFECTIVE DISEASES.
Dried diphtheritic membrane and cultures dried on silk threads
retain their vitality for several months.
A broth-culture in forty-eight hours may be used for inoculating
guinea-pigs. A few drops will cause death in from three to five
days ; there is hyperemia and edema at the seat of inoculation, the
lymphatic glands are enlarged, there is fluid in the peritoneal,
pleural and pericardial cavities, and the lungs are congested. The
bacillus is found at the seat of inoculation, but not, as a rule, in the
blood or internal organs. Inoculation of rabbits produces extensive
local cedema, enlargement of the lymphatic glands, and death in
from four days to three weeks. Roux and Yersin pointed out that in
less acute cases there was paralysis of the hind limbs. Mice and
rats have an immunity. Cultures lose their virulence with age, but
the filtrate from old cultures contains more toxic substance than
that from fresh cultures. The toxin has been described in a
previous chapter (p. 46).
Old cultures sterilised by heating for an hour to 60° C. or
70° C. will render guinea-pigs immune in two weeks. The toxic
substance is believed to be destroyed by this process, while according
to Frankel the immunity-giving substance which is also present in
the culture is not affected.
According to Behring’s researches, the blood of immune animals
contains diphtheria antitoxin, consequently the blood of an im-
mune animal is capable of neutralising the toxic properties in a
filtered culture, not only in the living animal but when added to the
culture in a test-tube. These researches led to the employment of
the serum of an immune animal as a therapeutic agent in the treat- ~
ment of diphtheria in man (p. 58).
Bacteriological Diagnosis.—The diphtheritic bacilli are not only
found in the throat while the lesions exist, but they are found after
all sign of the disease has disappeared. In some cases they
persist for a few days, in others for three or four weeks, and
in rarer cases they have been found several months afterwards.
They have also been found in the throats of persons in health,
especially of those who have been in contact with cases of diph-
theria, such as healthy children in infected families and healthy
nurses.
The bacilli which persist in the throat after recovery may be
virulent up to the time of their disappearance, or they may gradu-
ally become attenuated, and entirely lose their pathogenic properties.
The value of a microscopical examination as an aid in the diagnosis
of diphtheria has been considerably exaggerated, and unless the
a
‘ DIPHTHERIA. 335
bacillus when isolated is tested by inoculation the test may prove
to be entirely fallacious.
Léffler and Von Hoffman both found bacilli in healthy throats,
and thus created doubt as to the importance of the Liffler bacillus.
Hoffman found this bacillus in the throats of twenty-six out of
forty-five individuals, some of them suffering from scarlet fever,
measles or some other non-diphtheritic affections, while the rest were
healthy. The bacilli from these sources showed slight differences
in morphological and cultural characteristics, and Hoffman was
unable to decide whether these bacilli were diphtheria bacilli, which
had become harmless, or whether they were accidental epiphytes,
belonging to a closely allied but different species.
Roux and Yersin confirmed these observations. In a hospital
for children in Paris without any question of the existence of
diphtheria they found the so-called pseudo-diphtheria bacilli in
fifteen cases out of forty-five. In a school, in a seaside place
entirely free from diphtheria, the same bacilli were found in
twenty-six out of fifty-nine children. They were also found in-
children with simple sore throats, and in five out of seven cases
in measles. Roux and Yersin concluded'that these bacilli were
not distinct from the Léffler bacillus. There were slight variations,
but there was no constant difference except in their pathogenic
properties. The appearance of the colonies, the growth in broth,
and the peculiar morphological elements showed characters common
to both, and there was, in fact, less difference than there is
between attenuated anthrax and virulent anthrax in form and in
cultures; but inoculations of the bacillus did not cause death,
though in some cases in guinea-pigs there was marked cedema at
the seat of inoculation. On the other hand, Léffler’s bacilli
possess different degrees of virulence, some cultures producing only
temporary edema, while others cause death in twenty-four hours.
Virulent diphtheria bacilli subjected to a current of air can
in two weeks be deprived of their virulence partially, and in four
weeks entirely. Weakened bacilli can be raised in virulence by the
simultaneous injection of the streptococcus of erysipelas, but bacilli
deprived of their virulence and bacilli originally non-virulent cannot
be made to assume virulent properties. Escherich maintained
that they could be distinguished by comparative cultures; that the
pseudo-diphtheria bacilli made broth alkaline, so that in forty-eight
hours litmus was turned red by Léffler’s bacilli and blue by the
false bacilli. The bacilli themselves, according to Hoffman, are, as
a rule, shorter, wider and more uniform in size.
336 INFECTIVE DISEASES,
Parke and Beebe, with a view to clearing up this question, made
cultures from three hundred and thirty healthy throats. They
found bacilli of three varieties: bacilli characteristic in growth
producing acid reaction in broth, but having no virulence; bacilli
not characteristic in growth producing an alkaline reaction in broth,
not virulent; and bacilli producing acid reaction in broth and
virulent. The virulent characteristic diphtheria bacilli were found
in eight cases, non-virulent diphtheria bacilli in twenty-four, and
non-virulent false diphtheria bacilli in twenty-seven. They con-
cluded that the eight cases must have been in contact with diphtheria,
although the throats were healthy. With regard to the bacillus
in the twenty-four cases they regarded it as the true diphtheria
bacillus which had lost its virulence, and the bacillus found in the
twenty-seven cases showing differences in size and manner of staining
and the reaction produced in broth was properly designated pseudo-
diphtheria bacillus.
DipHtTHERitic DisEASES IN ANIMALS.
There are diptheritic diseases of the lower animals which are in
some respects similar to, and, some observers maintain, identical
with, human diphtheria.
In pigeons there is a disease accompanied with the formation of
false membranes associated with a bacillus described by Léffler.
Bacterium of Diphtheria of Pigeons (Bacillus columbarum,
Liffler).—Short rods with rounded ends, mostly in irregular masses.
In plate-cultivations on nutrient gelatine they formed whitish patches
on the surface, and compact, ball-like masses when embedded in the
gelatine. They were also cultivated on blood serum and potatoes.
Subcutaneous inoculation of a pure-cultivation produced in pigeons
local inflammation and necrosis ; inoculation in the mucous mem-
brane of the mouth gave the appearances of the original disease.
Other animals were only locally affected, except mice, in which
characteristic symptoms and death resulted. They were isolated
from the diphtheritic exudations in pigeons, and in sections were
found in the vessels of the lungs and liver.
A similar disease is known to attack fowls, and there are also
diseases with development of false membranes of the respiratory
passages in horses, cats and swine. Outbreaks of these diseases
have been said to occur in times of prevalence of diphtheria in
man, and their intercommunicability has been suggested.
Dr. Turner supposes that diphtheria in man originates in diseases
DIPHTHERITIC DISEASES IN ANIMALS. 337
simulating diphtheria in cats, pigs, and horses; and Klein, who
accepts this theory, maintains that cats suffer from genuine
diphtheria, and that after death the lungs are found full of grey,
consolidated lobular patches, and the kidneys are enlarged and
white.
Human diphtheritic membrane inoculated subcutaneously in cats
produces a painful swelling in the groin, and fever, and a fatal
termination in a week. The subcutaneous and muscular tissues at
the seat of.inoculation are hemorrhagic and edematous. The
internal organs are congested, and ‘in the kidneys the medulla is
congested, while the cortex is fatty.
A recent culture produces illness in twenty-four hours, a painful
tumour forms at the seat of inoculation, and death ensues in from
two days to a week. Pneumonia and fatty white kidney are found
after death, and the tissues at the seat of inoculation are hamor-
rhagic, and in parts almost gangrenous.
Klein found that diphtheritic membrane or a pure-culture in-
oculated into the cornea after removal of the superficial epithelium
produced ulceration, and in two cases perforation of the cornea,
and purulent panophthalmitis. Bacilli were again recovered from
the ulcer similar in cultural characters, but conspicuously shorter
and thinner.
An epidemic occurred amongst cats at the Brown Institution.
Five out of fourteen died. The symptoms were, running from the
eyes, sometimes a muco-purulent discharge, sneezing, coughing, and
pulmonary trouble, resulting in emaciation and death in from one
to three weeks. After death lobular pneumonia and large white
kidney were found; and in one case a diphtheritic condition of the
trachea, preparations of which showed diphtheria bacilli in crowds
under the microscope.
Klein regarded this disease as an epidemic of cat-diphtheria,
and believed that the disease was possibly induced accidentally by
the cats drinking milk, which was infected in the course of some
other experiments with diphtheria. He states that on account
of the very definite results obtained by inoculating diphtheritic
membrane and cultures of the bacillus, subcutaneously and on
the cornea, and of the condition of the lung and kidney in cats
naturally or experimentally infected, the disease must be con-
sidered as equivalent to human diphtheria, and the cat capable
of communicating the disease to other cats, and also to human
beings. These conclusions have not yet, met with the acceptance cf
veterinary authorities. The results of the experimental inoculations
22
338 INFECTIVE DISEASES.
are certainly by no means conclusive. It does not follow from these
experiments that the disease diphtheria naturally occurs in the
cat or that under ordinary circumstances cats may contract the
disease from the human subject; but the experiments show that,
like’ guinea-pigs and rabbits, cats are susceptible to the toxic effects
of the extremely poisonous principles developed during the growth
of Léffler’s bacillus.
Mitx DipHtTHeEria.
It has been shown that milk infected with diphtheria has been
the cause of epidemics among the consumers ; there have also been
epidemics apparently associated with the milk supply, in which it
hhas not been possible to trace the source from which the milk was
infected. A difficulty in tracing the origin in no way excludes the
possibility of contamination from a human source. In the light of
recent researches we should expect that it would be easy to overlook
the source of the virus, if it be true that diphtheria may exist without
any symptoms indicating its presence, and be unrecognised until the
throat has been examined for diphtheria bacilli. As this fact was
unknown until quite recently, the absence of an acknowledged
case of diphtheria was taken as evidence that no diphtheria existed,
and consequently that the milk must have been infected by a
diseased condition of the cow. Mr. Power, whose views upon milk
scarlatina have already been referred to, endeavoured to trace the
origin of a milk epidemic to the very common disease of “ garget,”
or mammary abscess. This idea may be dismissed without further
consideration ; but the theory of some disease existing in the cow
capable of producing diphtheria in man was resumed by Dr.
Cameron, who suggested that there might be an eruptive disease of
the teats producing diphtheria, and by Mr. Power, who supported the
theory in an investigation of a milk-diphtheria outbreak in 1886
at Camberley. Diphtheria in this case existed in the neighbourhood,
but as the source of human infection could not be traced, attention
was drawn to two cows in the herd which had recently calved, and
especially to one with chapped teats. Following this line of inquiry,
Klein investigated the behaviour of milch cows to the diphtheria
bacillus, Two cows were injected subcutaneously under the skin of
‘the shoulder with a Pravaz’ syringe filled with a sub-culture in broth.
There was a rise of temperature, and on the third day a painful
tumour, which enlarged to the size of a child’s head. In about a
fortnight the tumour began to decrease, and ultimately one cow
MILK DIPHTHERIA. 339
died and the other was killed. Such results might have been
anticipated as the result of injecting a large quantity of the toxic
products of the bacillus, but certain other phenomena were observed
to which importance was attached. On the fourth day, on one of
‘the cows an eruption on the teat was first noticed, consisting of
small vesicles passing into pustules and crusted ulcers. Examina-
tion of the contents of the vesicle revealed the bacillus. With
matter from the vesicles and pustules two calves were inoculated,
and a similar vesiculation produced at the seat of inoculation.
The milk of the cows was inoculated on nutrient gelatine, and
produced a culture of Bacillus diphtherie. The question naturally
arose whether this eruption had any connection with the original
experimental inoculation. No other cows in the locality from
which these cows were obtained had a similar eruption, and it was
taken for granted that it was the result of the experimental inocu-
lation. By accepting the possibility of this eruption being identical
with the chaps on the teats of the Camberley cows, the theory was
gradually built up that cows suffer from diphtheria, which manifests
itself in the form of an eruptive disease of the teats, and that the
disease is conveyed in the milk to the consumers. é
In the original experiment the bacilli were found to have
multiplied abundantly in the tumour at the seat of inoculation. The
eruption might have been, as admitted by Klein, a symptom of the
work ‘of the chemical poison, and the elimination of the bacilli by the
milk is also possible ; but that there is in cows a vesicular disease of
the teats which is the origin of human diphtheria is not accepted by
veterinarians, and there is not sufficient evidence to justify the con-
clusion that the infectivity of the milk in epidemics of milk diphtheria
has heen proved to be due to a morbid condition of the cow.
CHAPTER XXIV.
TYPHOID FEVER.
TypHorp Fever is a specific febrile disease peculiar to man, with
characteristic pathological lesions in the intestine, mesenteric
glands, and spleen. The Peyer's glands pass through three stages.
They become swollen from infiltration of round cells in lymph
follicles, due, it is supposed, to the presence of the typhoid fever
bacillus. The enlargement of the lymph follicles is followed by
coagulation necrosis until the entire patch becomes necrosed, and
sloughs away, leaving an ulcer. The disintegration of the patch
may extend in depth, and result in perforation and peritonitis, or
the ulcer may heal, and a pigmented scar take the place of the
Peyer’s patch. The lymphatic glands are found more or less
enlarged, and may be easily felt in the groin, axilla, and neck. In
some cases there is a tendency.to hemorrhage, followed by infarctions
in the spleen and lungs, which may develop into pyzmic abscesses
In the mesenteric glands similar changes take place, but without
ulceration. Pneumonia may occur as a pulmonary complication.
The bacteria of pneumonia and Streptococcus pyogenes may be
found in association with the bacillus of typhoid fever. It is
now generally accepted that the disease is conveyed by water and
food which have become contaminated with the virus contained
in typhoid evacuations. This has been practically proved by the
number of cases which have been shown to have been intimately
connected with contamination of drinking water from wells and
other sources, by sewers, cesspools and faulty drains, the sewage
presumably having been infected with typhoid excreta ; but whether
sewage independently of typhoid contamination can originate
typhoid is still an open question. Accepting the former theory
as a working hypothesis, we must assume that a typhoid fever
bacillus exists in the intestinal evacuations, and that it must be able
to retain its vitality under very varying conditions until it gains
340
TYPHOID FEVER. 341
access by the mouth to a fresh host, and by its development in the
intestine, and by the absorption of its toxic products, produces the
phenomena which we recognise as typhoid fever.
Fic. 129.—TypHor Fever. Iteum or ADULT, SHOWING SLOUGHY AND
INFILTRATED PatcHes (HaMItton).
Typhoid fever is also disseminated by milk ; sewage- contaminated
water having been added to the milk, or used for washing the milk
cans and other vessels,
342 INFECTIVE DISEASES.
Typhoid fever cannot be communicated to the lower animals.
Numerous experiments have been made by feeding and by injecting
typhoid stools, but with absolutely negative results. Murchison
gave typhoid fever discharges to pigs, Klein experimented with
rabbits, monkeys, and other animals. Motschutkowsky injected the
blood from cases of typhoid into monkeys, rabbits, and other animals,
but with negative results.
Fic. 130.—TyrHorw Bacitir rrom a CoLtony ON NUTRIENT GELATINE, x 1000
(FRANKEL AND PFEIFFER).
Various micro-organisms have been described in typhoid, but the
one to which most importance is attached is a bacillus which was
first discovered by Eberth, but cultivated and fully described by
Gaffky. Gaffky cultivated it from typhoid
evacuations, from typhoid ulcers, from
the mesenteric glands, and from the
p HEN Lin spleen. It is found in scattered colonies
in the spleen, and is rarely if ever present
in the blood.
Bacillus of Typhoid Fever.— Rods
1 to 3 in length, and ‘5 to ‘8 » in breadth, and threads (Plate VIIL.,
Fig. 2). Spore-formation has not been observed, but the protoplasm
may be broken up, producing appearances which may be mistaken
for spores. They are actively motile, and provided some with a single
and others with very numerous flagella, which are from three to five
times as long as the bacilli. They stain well with aqueous solutions
Fic. 131.—TypHorp Bactitit,
x 950 (BAUMGARTEN).
TYPHOID FEVER. 343
of aniline dyes, and grow well at the temperature of the room. In
plate-cultivations minute colonies are visible in thirty-six to forty-
eight hours ; they are circular or oval, with an irregular margin ; they
appear granular by transmitted light, and are yellowish-brown in
colour. Cultivated in the depth of gelatine a well-defined shiny film
forms at the point of puncture, and a greyish-white filament, com-
posed of closely packed colonies, develops in the track of the needle
(Fig. 134). On the surface of gelatine a greyish-white translucent
film forms, with sharply defined margin (Plate IT., Fig. 2). On agar
there is a whitish transparent layer. They flourish in milk. On
potato at the temperature of the blood there is no culture visible, but
Fig. 132.—FLAGELLA OF TypHoID BacrLui1, x 1000, STAINED BY LOFFLER’s
Metuon (FRANKEL AND PYkEIFFER).
the inoculated area appears moist and shining, and cover-glass pre-
parations made from the potato will demonstrate that there is really
a copious growth of the bacillus. This almost invisible growth is not
peculiar to this micro-organism.
Whether this bacillus is really peculiar to typhoid is much dis-
puted. Bacilli very closely resembling it, if not actually identical,
have been found under other conditions. These pseudo-typhoid bacilli
are regarded by some bacteriologists as varieties resulting from the
different environment afforded by a saprophytic existence. Numer-
ous experiments have been made on animals with pure-cultures of
the bacillus, but in the production of typhoid fever they have
been no more successful than the experiments with typhoid stools.
344 INFECTIVE DISEASES.
Frankel and Simmonds inoculated a number of rabbits in the vein of
the ear, producing death, in some cases in forty-eight hours.
Seitz
administered broth-cultures by Koch’s method of introducing them
Fic. 133.—Cotonies or TypHorp Bacituus.
Three days old. x 100 (FRANKEL AND PFEIFFER).
cultivations, a similar result following the
injection of sterilised cultures. An account of
_the products has already been given (p. 41).
Cassedebat isolated three species of bacilli
from water, which could be distinguished
with great difficulty, and only after the most
The bacillus which most
closely resembles it is the Bacillus coli com-
munis; in fact, Roux regards it as a non-
pathogenic variety of the typhoid bacillus.
Others claim to be able to distinguish it by
careful comparison and the application of
tests. Special importance is attached to
potato cultures, the typhoid bacillus forming
an invisible film, and Bacillus coli communis
a well-marked yellowish growth. Terni
pointed out that Bacillus typhosus retains its
motility in media containing hydrochloric
careful comparison.
into thestomach after
the administration of
opium in guinea-pigs,
and death resulted in
several instances.
But in all these cases
the results depended
upon the poisonous
products found in the
Fic. 134.—Pure-CuLtTure
or TypHoID BactLur
INOCULATED IN THE
DrrtH oF NUTRIENT
GELATINE (BAUMGAR-
TEN).
acid, while Bacillus coli communis and other bacilli resembling those
of typhoid lost their motility. Schild maintained that Bacillus typhosus
TYPHOID FEVER. 345
was destroyed by exposure to the vapour of formalin, while Bacillus
coli communis and similar bacilli isolated from water gave subcul-
tures after exposure for two hours. Typhoid bacilli do not give the
reaction for indol, and there is no development of gas in cultures in
the depth of nutrient agar containing 2 per cent. of grape-sugar.
According to Miiller, sterilised milk is coagulated in twenty-four
hours, at 37° C., by Bacillus coli communis, but not by the Bacillus
typhosus until several weeks have elapsed ; and, further, cultures on
acid potato give different results. The typhoid bacillus on micro-
scopical examination shows marked polar staining, but Bacillus
coli communis only shows a slight tendency of the protoplasm to
break up.
¥ic. 135.—TypHorp BAcILLI In A SEcTION oF SPLEEN, x 800 (FLUcGE).
Kitasato suggested the negative indol test, and recommended
Salkowski’s method. Broth-cultivations are treated with a solution of
sodium or potassium nitrite: 1 cc. of the nitrite solution (-02 gr. to
100 ce. of water) is added to 10 ce. of a broth-culture after twenty-
four hours in the incubator, and on adding a few drops of strong
sulphuric acid, the typhoid cultures remain colourless, but cultures
of bacilli apparently identical give the red colour. On the other
hand, Losener maintains that he has cultivated from earth, water
and healthy human evacuations bacilli which could not be distin-
guished from typhoid bacilli by any of these tests.
The detection of the typhoid bacillus in water has been
346 INFECTIVE DISEASES.
described in another chapter (p. 147) ; but sufficient has been said to
show that bacteriological reports in which it is stated that the typhoid
fever bacillus has been found in water causing typhoid epidemics
must be accepted with great reserve ; and further, no one is justified
in stating that the typhoid fever bacillus is undoubtedly the cause
of typhoid fever. It is not found in every case of typhoid, it is
not found in the blood, but it is found in those tissues which are
Fic. 136.—TypHom Bacitii in A SECTION OF INTESTINE, INVADING THE SUBMUCOUS
(T.J.) anD Muscutar Layers (M.), x 950 (BAUMGARTEN).
e
commonly the seat of secondary invasion of epiphytic bacteria,
whose normal habitat is the intestinal canal.
Lastly, as the disease does not exist in the lower animals, the
crucial test cannot be applied. The etiology of typhoid fever is
still enveloped in doubt, and the nature of the contagium has not
yet been determined.
CHAPTER XXV.
SWINE FEVER.
Pie TypHorp, or swine fever, is a highly contagious disease peculiar
to swine, causing death in from ten to thirty days, associated with
a fibrinous pneumonia, enlargement of, and hemorrhage into, the
lymphatic glands, and characteristic ulcers of the mucous membrane
of the stomach and intestines. The lesions may assume the form of
extensive croupous or diphtheritic deposit, which may fill the intes-
tinal tube. But the most characteristic appearance results when
the lower part of the ileum and commencement of the colon is
dotted all over with elevations of the mucous membrane, resembling
leather buttons or nux vomica seeds, and sometimes with concentric
rings, so that they have been compared to slices of calumba root.
Swine fever is difficult to detect in the early stage, and sometimes
symptoms are absent altogether in animals suffering from the
disease and quite capable of transmitting it; or nothing may be
noted except cough, and possibly enlargement of the inguinal glands.
In typical cases the animals are noticed not to feed, to exhibit
dulness, and to have occasional rigors. Partial paralysis may
follow, producing unsteady gait or loss of power over the hind legs.
Diarrhea sets in, and the evacuations become blood stained. There
is occasionally a diffused or patchy reddish or purplish rash on
the skin. After death the appearances most commonly found are
inflammation of the peritoneum, and redness and enlargement of the
mesenteric glands and the lymphatic glands in the lungs. There
is generally ulceration, especially of the colon and ileo-czcal valve,
or a diphtheritic exudation, sometimes pale yellow, more commonly
greyish or black, similar to the centres of necrosis within the ulcers.
The spleen is enlarged and liver congested, and there are hemorrhages
in the kidneys. As the lungs are so commonly affected, Klem
proposed the name pneumo-enteritis; but the pulmonary lesions
are not constant. Indeed, the cases in which the intestines and
~ BAT
348 INFECTIVE DISEASES.
lungs are simultaneously affected are not numerous, and sometimes
the lungs may be found to be perfectly healthy in cases with
Fic. 137.—ULcrraTioN OF THE INTESTINE IN A TYPICAL CASE OF SWINE-FEVER.
long-standing lesion of the intestine. Old pigs may linger on for
weeks, and ultimately recover, and in the meantime act as centres
for the dissemination of the disease.
DESCRIPTION OF PLATES IX. AND X.
Swine Fever.
Puate IX.—Part of intestine from a typical case of swine fever, showing
scattered ulcers and ulceration of the ileo-cxecal valve.
PLATE X.—From the same case of swine fever. The lungs were extensively
inflamed and partly consolidated, and the lymphatic glands were enlarged
and of a deep red or reddish-purple colour.
Plate IX.
SWINE FEVER.
Vincent Brooks,Day & Son, lath.
| Crockshanke fecit is
Plate X.
SWINE FEVER.
E. Crackechante foci? Vincent Brooks,Day & Son, lith.
SWINE FEVER. 349
Budd first pointed out that this disease might be compared
to human typhoid, both diseases being attended by a peculiar
ulceration of the intestinal follicles; but the diseases are not to
be considered in any sense identical or interchangeable.
Bacteria in Swine Fever.
—In 1877 Klein published a
research in a Report to the Local
Government Board, in which L y) Wie
he claimed to have discovered _ o fs 8
bacilli characteristic of the a y
—<—
disease. They were described as
similar to Bacillus subtilis, or
Iie. 138.—Bacitius or SwINe-FEVER
201 <r > “1 }
Bacillus anthracis, but smaller Riouee acne)
in size. These bacilli developed
into long leptothrix filaments, and formed spores. It was further
asserted that on inoculation, cultures produced lesions indicative of
swine fever; the bacilli were also pathogenic in mice and rabbits.
. = Later this bacillus was re-
, een \ 4 nounced in favour of another.
= | pe’ <= In the following year Det-
\ \ Ze mers described a bacillus, but
\ J AEN ,. & subsequently renounced it in
x : favour of a micrococcus.
aa ¥ ‘ fap | In 1882 Pasteur maintained
eS ] S23 al that the virus of swine fever in
os France (rouget) was a dumb-bell
Fic. 139.—Bacmtus No. 2. From a micrococcus, which produced the
PREPARATION OF BRONCHIAL Mucus
same effect in pigeons as the
or A Pic. (KuxErn.) pig :
microbe of fowl-cholera. Though
rouget or swine measles is probably a different disease, the occurrence
of this micro-organism is of interest in this connection.
In 1883 Klein again investigated swine fever, and discovered
Bacillus No. 2, and maintained
that these bacilli were found in
the blood, in the peritoneal and
bronchial exudations ; and in the
air vesicles of the lungs, in the
form of leptothrix filaments ten
Fre. 140.—Bacititus No. 2. From an
or twenty times the length of Lamm: Cumin (cai)
single rods. Cultivations were
made on solid media. The organisms in these cultures were minute
rods actively motile, occurring singly or forming chains, two or three
350 INFECTIVE DISEASES.
times as long as Bacterium termo; and in preparations made from
diseased organs they were found to possess a very narrow trans-
parent halo, a sort of hyaline gelatinous capsule, Inoculation of
cultures failed to produce the lesions found in animals naturally
infected. Two pigs were inoculated, one with a sub-culture from the
swollen bronchial gland of a pig that had died of pig-typhoid, and
a second with a culture obtained from the spleen of a mouse that
had been inoculated from another case of swine fever. After two
days the inguinal glands near the seat of inoculation became swollen,
and the temperature rose slightly, After three or four weeks the
ion recovered.
Mice on the fifth or sixth day after inoculation showed symptoms
of illness, then respiration became
superficial and slow. Death
occurred on the sixth or seventh
ay
day.
Rabbits showed a rise of
temperature, and death followed
between the fifth and eighth days,
the temperature falling before
death. At the post-mortem ex-
amination there was usually
peritonitis, with copious exuda-
tion, The kidney, spleen, and
a Mouss, arrer Inocunation witn liver were enlarged and dark in
Bacitius No. 2. (KiE1y.) many cases, there was red hepa-
tisation of the lobes of the lungs,
and generally pericarditis and hemorrhage under the pericardium.
In 1885 Salmon, in the annual report of the United States
Bureau of Animal Industry, published the result of his investigations
into American hog cholera, which is identical with English pig typhoid.
A motile figure-of-eight bacterium was isolated, each part being
about twice as long as broad. The bacterium grew on nutrient
gelatine without liquefying it, and on potato produced a brownish
growth; broth tubes became turbid on the following day. Colonies
in plate-cultivations were oval or circular, and brownish in colour.
Six pigs inoculated subcutaneously were all said to have died of hog
cholera, and the bacterium was again obtained from the blood of
the heart and spleen. The bacteria proved fatal to mice, rabbits,
guinea-pigs, and pigeons.
In 1893 Welch and Clement described the hog-cholera bacillus
as variable in form, and they further stated that a culture obtained
Fic. 141.—Bioop or FresH SPLEEN OF
SWINE FEVER. 351
from Klein, while not possessing the characters originally ascribed
to it, could not in its form, biological characters, or pathogenic pro-
perties, be distinguished from the American hog-cholera bacillus.
In 1887 pig typhoid was investigated at Marseilles by Rietsch,
Jobert, and Martinaud, and a bacillus found. This grew rapidly
on all the nutrient media. In gelatine a growth was obtained in
twenty-four hours at 18° C.; on blood serum and agar an opaque
growth developed; and on potato the growth was yellowish. It
was asserted that 'a young pig was killed by a culture in twenty-
two days, and that the characteristic ulcerations were observed in
the intestines,
In 1887 pig typhoid was prevalent in Sweden, and Bang and
Selander experimented with cultures from a rabbit that died after
inoculation with a fragment of spleen from a diseased pig. The
bacilli were motile, varying from rods to cocci, without spore-formation
and pathogenic in mice, guinea-pigs, and rabbits, but not in pigeons.
Pigs fed on broth-cultures were said to have succumbed to genuine
pig typhoid. In the blood they were generally found in the form
of short oval bacteria, but in the blood of the heart longer rods
were sometimes found. Metchnikoff described a bacillus isolated by
’ ‘Chantemesse from an outbreak in France, as highly polymorphic.
Smith identified the hog-cholera bacillus with the bacillus found
by Schiitz, and this in turn has been identified with the bacillus
of hemorrhagic septicemia.
From these researches it would appear to be probable that one of
the bacteria isolated by Klein, and those found by Salmon, Smith,
Bang, Welch and Schiitz are identical; and further, that they have
been identified with the bacillus of hemorrhagic septicemia. We
may sum up the characters thus :—
Bacillus of Klein, Salmon, Smith and Schiitz.—Very small
rods, actively motile ; spore-formation not observed ; colonies circular
and brown by transmitted light. Inoculated in the depth of gelatine
faintly yellowish-white colonies develop along the track of the needle ;
‘on the surface an opalescent film; on potato they produce a straw-
coloured layer, changing to brown. There is absence of indol in
cultures containing peptone; the bacilli are fatal to mice, guinea-
pigs, rabbits, and pigeons. Swine die after intravenous injection,
but not, as a rule, after subcutaneous injection. .
According to Caneva, the bacillus obtained from the Marseilles
epidemic would appear to be closely allied, if not identical, with
the bacillus of ferret disease (Eberth and Schimmelbusch), and the
bacillus of Texas fever (Billings).
352 INFECTIVE DISEASES.
Billings appears to have isolated two bacilli, one identical with
the Marseilles bacillus, and the other with the hog-cholera bacillus.
Bacillus of Rietsch and Jobert.—Rods about twice as
long as broad, rather shorter than the bacillus of typhoid fever,
longer and thicker than the American bacillus. They exhibit
end-staining. They possess flagella, and are actively motile.
They grow rapidly in nutrient media. They are only feebly
pathogenic, ‘They are also said to be distinguished from the bacilli
of hog-cholera by producing indol in solutions containing peptone,
and by causing an acid reaction in milk.
Bacillus of M‘Fadyean.—M‘Fadyean investigated swine fever
in 1895, and found bacilli which he differentiated from hog-cholera
bacilli. The method employed was to inoculate the surface of
nutrient agar and of potato, with fragments torn out of the centre
of a lymphatic gland, with specially constructed forceps. Inocula-
tions were also made from the spleen pulp, and blood, in the usual
way. They are from 1 to 2 yw in length, and ‘6 mw in breadth.
They never grow into filaments, they do not form spores, and
they are actively motile. They are readily stained by the watery
solutions of the aniline dyes, and are decolorised by Gram’s
method; with methylene-blue they show end-staining. The bacilli
grow on gelatine without liquefaction, forming a thin white line
along the needle track. On agar a thin, transparent pellicle forms,
which is not easily visible at first, but gradually acquires a faint
greyish tint. More characteristic appearances result in plate-
cultivations of gelatine-agar, at 37°C. The colonies are distinctly
visible in eighteen hours, appearing when viewed by transmitted
light as bluish-white, circular specks ; each colony has a dark centre
and a granular margin. In broth the bacilli produce turbidity after
twenty-four hours. On potato there is no visible growth, even when
the surface is inoculated with an abundance of material. On solid
blood serum the growth is scanty. They grow in milk without
producing coagulation. They are harmless to guinea-pigs and feebly
pathogenic to rabbits.
Several experiments were carried out upon swine. In the first,
series the most rigid precautions were taken to prevent accidental
infection with swine fever. Four young pigs were inoculated, upon
a farm where there was no previous history of the disease. These
pigs were killed, and the post-mortem examinations were said to
show indications of swine fever, principally patches of diphtheritic
material in the colon, and healing ulcers. The next series of pigs
was inoculated at the Royal Veterinary College. Cultures were
SWINE FEVER. 353
administered with milk to two pigs. Five days afterwards one pig
died; and the mesenteric glands were congested, and the mucous
membrane showed spots of necrosis. The other pig was killed, and
there were ulcers in the colon. Jn a third experiment, eight pigs
were fed with milk and broth cultures. These pigs were all killed
at different dates, and most of them had ulceration of the colon ;
in control experiments the intestine was normal.
M‘Fadyean compared his bacillus with a culture of the hog-
cholera bacillus, and found that the American organism grows at a
lower temperature in gelatine, and colonies appear in plates much
earlier. They produce a less transparent and thicker growth, and
much greater turbidity in broth and a more abundant sediment.
On potato they form an abundant growth at 37° C., at first
yellow, later brown, with considerable resemblance to a glanders
culture.
Colonies upon gelatine-agar are distinguished by their opacity
and sharp outline. Agar, potato, and broth cultures of the American
organism consist of short ovoid forms like the bacilli of fowl cholera,
while the bacillus isolated by M‘Fadyean has a closer resemblance
to the bacillus of glanders. M‘Fadyean asserts that the American
organism is not pathogenic to the pig. Pigs after feeding on broth
cultures remained healthy, and showed no trace of swine fever when
killed from one to three weeks afterwards. On the other hand,
broth cultures of his bacillus produced the characteristic ulceration
of the bowel. M‘Fadyean claims, therefore, to have discovered the
true pathogenic organism of swine fever. He does not appear to
have compared this bacillus with that obtained from the epidemic
of swine fever at Marseilles. From the description of the mor-
phological and other details there seems to be a close resemblance
between the two.
Not less than three and possibly four species of bacilli have been
cultivated from swine fever, two at different times by Klein, one
by Reitsch Jobert and Martinaud, and one by M‘Fadyean ; and
cultures of all these bacilli have been credited with producing swine
fever in experimental animals, and each one has been pronounced
to be the contagium of the disease. We must conclude either that
contaminated cultures were inoculated in some cases, or, what is
far more probable, the swine fever which resulted in experimental
animals was due to accidental infection; and until a bacillus has
been cultivated from swine fever from which another investigator
can prepare sub-cultures, and with those sub-cultures produce the
typical ulcerations of swine fever in pigs on a farm, or on premises
23
354 INFECTIVE DISEASES.
in which swine fever is unknown, we are justified in concluding that
the contagium has not yet been discovered.
The mistakes which are likely to occur when the same investi-
gator isolates bacilli from cases of swine fever, and subsequently
inoculates or feeds healthy swine, cannot be better illustrated than
by quoting from a leaflet issued by the Board of Agriculture,
pointing out the great precautions necessary to prevent accidental
infection.
“There seems reason to believe that the disease is not infre-
quently introduced by means of persons who have been in contact
with diseased animals. Pig owners, therefore, are advised to prevent
strangers from at any time approaching their pigs, and should the
admission to the premises of spayers or castrators be necessary,
those persons should be required, before approaching the animals, to
thoroughly wash their hands with soap and water, and to wash and
disinfect their boots with a solution of carbolic acid and water, or
some other suitable disinfectant. Such persons might also with
advantage be required to wear, while operating, a waterproof apron,
which should be washed and disinfected before the wearer is per-
mitted to approach the animals to be operated on.”
Protective Inoculation.—The experiments of Salmon and of
Schweinitz have been referred to in another chapter (pp. 41, 46).
A method of protective inoculation was attempted in America, but
the experiments were unsuccessful, and the plan was abandoned.
Stamping-out System.—Notification is compulsory, and the
order in force is the Swine Fever Order of 1896, but the stamping-
out system has not been applied ina thoroughly satisfactory manner,
and the disease is still very prevalent.
CHAPTER XXVI.
SWINE MEASLES.—DISTEMPER IN DOGS.—EPIDEMIC DISEASE OF
FERRETS.—EPIDEMIC DISEASE OF MICE.
Swine MEAsLEs.
Swine MEAsLes, or swine erysipelas, is described as an acute, in-
fectious disease of swine which is very prevalent in France and
Germany, but is included in this country in the term “swine fever.”
According to some, it is a distinct disease, and distinguished from pig-
typhoid by absence of the ulceration of the intestines which is so
characteristic of that disease ; while, according to others, ulceration
of the intestine and ileo-cwcal valve may be found post-mortem. The
onset of the symptoms, as in pig typhoid, is very rapid; the animals
cease to feed, and show other general signs of illness; the voice is
hoarse, and there is a rapid rise of temperature. On the neck,
chest, and abdomen, red patches make their appearance, which
extend and coalesce, and change to a dark reddish or brownish colour.
These symptoms may be followed by convulsions, and sometimes by
paralysis of the hind legs; and death occurs in from one to four
days. It is especially a disease of young pigs, and from 50 to 60
per cent. of infected animals die.
On post-mortem examination there is hemorrhage and cedema
in the patches of the skin, the lymphatic glands are swollen and
dark red, the peritoneum is ecchymosed, the intestinal mucous
membrane is congested and swollen, and the solitary follicles and
Peyer’s patches are prominent, and in the neighbourhood of the ileo-
cecal valve there are, according to Fliigge, ulcers of considerable size.
The liver and spleen are congested and enlarged. Pasteur investi-
gated swine measles or rouget, and described a figure-of-eight micro-
coceus, which he believed to be the contagium of this disease. This
organism appears to be identical with the bacterium of hemorrhagic
septicemia, which is also commonly found in pig-typhoid.
In experimenting with the virus obtained from the spleen
365
356 INFECTIVE DISEASES.
Pasteur found that, by successive inoculation of rabbits, the virulence
was exalted for rabbits, but attenuated for swine, and the virus
which had thus been passed through the rabbit was used as a
vaccine for swine, to protect them against virulent erysipelas.
Pasteur found that by passing the virus through pigeons it was
made more virulent for swine.
In the blood, and the juice of the internal organs, and of the
lymph glands, Schiitz found a minute bacillus identical with the
bacillus of mouse septicaemia.
Bacillus of Swine Erysipelas (Schiitz)—_Hxtremely minute
rods ‘6 to 1°8 » in length, morphologically and in cultural charac-
ters identical with the bacillus of mouse septicemia. Filaments
and involution forms. Spore-formation present.
House mice if inoculated with a pure culture die in two to four
days. Pigeons are also very susceptible. Fowls and guinea-pigs
i/
Nts
Me
Apri byline
\
Fic. 142.—Bactnt or Swink Fic. 143.—Bioop or PicHoN INOCULATED
ERYSIPELAS (BAUMGARTEN). WITH BaciLii or Swine ERysIPELas, x 600
(ScutTz).
are immune. Rabbits after imoculation of the ear suffer from
erysipelatous inflammation, identical with that produced by inocu-
lation of the bacillus of mouse septicemia. The bacilli are also
pathogenic in swine and sheep.
Protective Inoculation.—With Pasteur’s vaccine immunity is
said to be produced which lasts about a year. Schiitz and Schottelius
found the minute bacilliin Pasteur’s vaccine, which they had already
found in cases of swine erysipelas in Germany.
The results of vaccination in France are said to be very satis-
factory, but m test experiments in Germany they were not so
favourable. Out of 119 vaccinated swine 5 per cent. died as the
result of the inoculation, while the average loss in the ordinary way
is 2 per cent.
Metchnikoff found that the blood of immunised rabbits was
antitoxic, and Lorenz maintains that the serum of swine which
SWINE FEVER. 357
have recovered from swine erysipelas is also antitoxic, and will
produce immunity in other animals. The treatment introduced
by Lorenz is to inject serum in the proportion of 1 cc. to every
10 kilogrammes of the weight of the animal's body. Two days
afterwards -5 to 1 cc. of virulent culture is injected, and after
twelve days the dose is doubled. Lorenz inoculated 294 pigs;
12 were suffering from swine erysipelas, and
of these 6 recovered and 6 died.
In the opinion of the author this disease
requires re-investigation, for if it be true that
rouget or schweinrothlauf is associated with
ulceration of the intestines, the recognition
of it as a disease distinct from our English
swine fever apparently rests upon the pres-
ence of a bacillus, which cannot be distin-
guished from the bacillus of mouse septiccemia.
The question arises whether this bacillus
is really the cause of a distinct disease, swine
erysipelas, or, on the other hand, whether
the bacillus is really the bacillus of mouse
septicemia which has been isolated from
certain cases of swine fever. The bacillus of
mouse septicemia is widely distributed, and
it may only be an accidental concomitant in
rouget or schweinrothlauf. The presence of
the bacterium of hemorrhagic septicemia in
both rouget and pig typhoid would not prove
identity, as this micro-organism is un-
doubtedly only secondary in both diseases.
There is great need, therefore, for further Pye. 144.—Purn-cuLruRE
careful investigation. Clinical and patho- 1x Nurrrent GeELa-
TINE OF BACILLI
R In Swine ERyYSIPELAS
country, to determine whether there are (BAUMGARTEN).
really two diseases included under the name
“swine fever.” If this prove to be the case, we must ascertain the
clinical and pathological differences between rouget and pig typhoid.
How can rouget be distinguished from cases of swine fever in which
there is a patchy rash, paralysis of hind legs, but no ulceration of the
intestine 2 Further, how is swine erysipelas with ulceration of the
intestine and ileo-ccecal valve to be distinguished from an ordinary
logical observations must be made in this
case of pig typhoid ?
358 INFECTIVE DISEASES.
Distemper In Does.
Distemper is an infectious febrile disease of dogs, characterised by
bronchial catarrh and discharge from the eyes. Bronchitis and
pneumonia may supervene, or there may be intestinal catarrh ter-
minating in dysenteric diarrhea, sometimes complicated by jaundice,
The disease may affect the nervous system, and produce convulsive
contractions of the muscles of the nose, ears, lips, and limbs.
Occasionally there is an eruption, especially in animals which are
out of condition. The virus exists in the discharge from the nostrils
and eyes, and is given off from the lungs and the skin.
One attack of the disease does not confer entire immunity ;
and some dogs are completely insusceptible.
Bacteria in Distemper.—Millais has isolated a micro-organism
resembling the pneumococcus of Friedlander, which he believes to be
the cause of the disease. The bacillus occurs with other bacteria and
micrococci in the nasal discharge.
Protective Inoculation.—Mixed cultures of these bacteria
liquefy the gelatine, and the liquid has been used as a vaccine.
When applied to the nose, it is said to produce a mild attack of
distemper, which protects as much as an attack of the disease
contracted naturally. These results require confirmation.
Inoculation of the nasal discharge in healthy dogs has been
practised, so that they may have the disease under favourable con-
ditions ; but the system should not be encouraged, as dogs need not
necessarily contract distemper. Vaccination with cow-pox lymph
has been advocated, but it is perfectly useless.
Stamping-out System.—Dogs suffering from distemper must be
completely isolated. Any straw or litter which has been in contact
with a diseased dog should be burnt. Clothing, collars, chains, and
the kennel or premises inhabited, must be thoroughly disinfected.
The animal after recovery should be washed with carbolic soap.
Eripemic Disease or FERRETS.
Eberth and Schimmelbusch investigated an epidemic disease of
ferrets (frettchen-seuche), and isolated a bacillus, which in mor-
phology and cultivation agrees very closely with the bacillus of
hemorrhagic septicemia.
Eripemic Disease or Mice.
Lofler investigated an epidemic disease which occurred in mice
kept in confinement, and isolated a bacillus resembling Bacillus
typhosus.
EPIDEMIC DISEASE OF MICE. 359
Bacillus Typhi Murium.—Rods varying in length; and fila-
ments; motile; flagellated. The colonies are circular, brownish
and granular on the surface of obliquely solidified gelatine. The
bacteria inoculated on the surface produce a greyish-white semi-
transparent growth, and on agar and potato the appearance of the
growth is very similar. They can be cultivated readily in milk and
in broth. White and field mice are killed in from one to two
weeks, when given bread moistened with a culture.
Liffier claims to have used this method with success in Thessaly,
where there was a plague of field mice causing great losses to
agriculturists.
CHAPTER XXVII.
ASIATIC CHOLERA. — CHOLERA NOSTRAS. — CHOLERAIC DIARRH@A
FROM MEAT-POISONING.—DYSENTERY.—CHOLERAIC DIARRHGA
IN FOWLS.
ASIATIC CHOLERA.
THERE are several diseases in man associated with diarrhcea, which
have certain characters in common, but are totally distinct. They
include Asiatic cholera, cholera nostras, dysentery, and choleraic
diarrhea. Asiatic cholera is an endemic disease of the Delta of
the Ganges, a locality which has become notorious as the home
of cholera. Cholera is a filth disease; and the accumulation of
filth on the banks of the Ganges, with contamination of the
water, and the climate, afford most favourable conditions for the
development of the cholera virus.
Four great cholera epidemics have originated in, and spread
from, India: in 1817, in 1826, in 1846, and in 1865. Cholera
follows the routes of pilgrims and caravans, and now, owing to
the rapid means of communication by steamers and railways, it
spreads to the most distant parts of the world, covering in a few
weeks or days distances which in former times could only be traversed
in several months or even years.
In 1892 the epidemic passed from India, through Afghanistan,
to Russia in Asia, and quickly spread westwards along the route
of the trans-Caspian railway ; and all this occurred within the space
of a few weeks. By Russian emigrants it was carried to Hamburg
and Antwerp; and the virus, finding a suitable environment in the
former place, produced a severe epidemic there. Thus, in about three
months, it was brought into close proximity with England. Mecca
is one of the great infective centres of the world, for there all
the conditions are found for the propagation of cholera, including
filth, overcrowding, and the water of the famous Holy Well, which
is used for ablutions and drinking purposes. The return of
360
ASIATIC CHOLERA. 361
the pilgrims to Egypt, and the proximity of England to Egypt,
necessitate the greatest possible precautions to prevent the intro-
duction of the disease into this country.
In 1884 a German Commission was sent out to India, and Koch
discovered a micro-organism which he described as a curved or
comma-shaped bacillus, and pronounced to be the contagium of
this disease.
Fic. 145.—Cover-cLass Preparation or A Dror or Meat INFUSION, containing
a pure-cultivation of comma-bacilli, with (a) spirilliform threads, x 600. (Koc#.)
Spirillum cholerz Asiaticze (Comma-bacillus, Koch).—Curved
rods, spirilla, and threads. The curved rods or commas are about
half the length of a tubercle-bacillus. They occur isolated, or
attached to each other forming S-shaped organisms or longer
screw-forms, the latter resembling the spirilla of relapsing fever.
a pfs ~25
Ce § Lie fe
8 ad S oe 080 20) x eter
GR 88 BE % me te
7 oul f
Fic. 146.—ARTHROSPORES ; (c) Comma-bacillus breaking up into spheres; (b, ¢),
formation of spheres in spiral forms; (d, e), groups of spheres; (f) spirilla
with spheres from an old cultivation; (yg) germination of the spheres.
(HvEppE.)
Finally they may develop into spirilliform threads. In old cultiva-
tions threads are found with swellings or irregularities (Fig. 148).
The commas are actively motile, and possess flagella (Fig. 147).
Their movements) and development into spirilla may be studied in
drop-cultivations. Arthrospore formation has been described by
Hueppe (Fig. 146). In plate-cultivations, at a temperature of from
362 INFECTIVE DISEASES.
16° to 20° C., the colonies develop as little specks, which begin to
be visible after about twenty-four hours. Examined with a low
power, and a small diaphragm, these colonies have the following
characteristics. They appear as little masses, granular, and
Fic. 147.—FLAGELLA OF COMMA-BACILLI; STAINED BY LOFFLER’S MElHon
(FRANKEL AND PFEIFFER).
yellowish-white in colour, and sometimes very faintly tinged with
red, which have liquefied the gelatine, and sunk down to the bottom
of the resulting excavations. :
In test-tubes of slightly alkaline nutrient gelatine (10 per cent.),
Fic. 148.—Invotution Forms, x 700 Fic. 149.—CoLonres Or COMMA-BACILLI
(Van ERMENGEm). on Nutrient GELATINE, NATURAL
Size (Kocw).
the appearance of the growth is very striking. In typical cultures
it begins to be visible in about twenty-four hours. Liquefaction
sets in very slowly, commencing at the top of the needle track
ASIATIC CHOLERA. 363
around an enclosed bubble of air, and forming a funnel continuous
with the lower part of the growth; the latter preserves for several
days its resemblance to a white
thread (Plate II., Fig. 1). In
about eight days, however, lique-
faction takes place along the
whole of the needle track.
On the surface of agar-agar
the cultivation develops as a Fic. 150.—Cotonres or Kocn’s Comma-
BACILLI, x 60.
white, semi-transparent layer,
with well-defined margin. The
appearance on blood serum is very similar ; liquefaction very slowly
takes place. In broth they form a wrinkled film on the surface,
there is a rapid and abundant growth at the temperature of the
WT \
iG) 4
tile oe
Nl Maram
Wi (HITS a]
Hh} | PS
{| ‘
; /
a NG My
TaN
Fic. 151.—Cover-ciass Preparation Fic. 152.—Cover-cLass PREPARATION
FROM THE CONTENTS OF A CHOLERA or CHOLERA DrJEcTA oN Damp LinEN
INTESTINE, x 600. (a) Remains of the (two days old), x 600. Great prolife-
epithelial cells ; (6) Comma-bacillus ; ration of the bacilli with spirilla (a)
(ec) Group of comma-hacilli (Koch). (Koch).
blood, and the same applies to sterilised milk; and they will even
multiply in sterilised water. In potato-cultivations the microbe
will only grow at the temperature of the blood (37° C.), forming a
slightly brown, transparent layer. Inoculation of a cultivation of
the bacillus in the duodenum of guinea-pigs, with and without
364 INFECTIVE DISEASES.
ligation of the bile-duct, has given positive results. More recently
these results have been confirmed by the following method: Five
ce. of a 5 per cent. solution of potash were injected into the
stomach of a guinea-pig, and twenty minutes after, 10 cc. of a
cultivation of comma-bacilli, diffused in broth, were similarly intro-
duced. Simultaneously with the latter, an injection of tincture of
opium was made into the abdominal cavity, in the proportion of
1 cc. for every 200 grammes weight of the animal. Those who
have had success with inoculation experiments maintain that choleraic
symptoms were produced without any trace of peritonitis or putrid
infection, and that the comma-bacilli of Koch were again found
in the intestinal contents, and fresh cultivations established.
Fic. 153.—SecTIon or THE Mucous MEMBRANE OF A CHOLERA INTESTINE, x 600.
A tubular gland (a) is divided transversely ; in its interior (b) and between
the epithelium and the basement membrane (c) are numerous comma-bacilli
(Koch).
On the other hand, these results have been disputed, the fatal
effects of the inoculation attributed to septiceemic poisoning, and
the proliferation of the bacilli considered to be dependent upon an
abnormal condition of the intestines, induced by the injection of
tincture of opium. It has, however, been shown that these organisms,
like several others which have been isolated from intestinal dis-
charges, produce definite poisonous substances. The comma-bacilli
were found in the superficial necrosed layer of the intestine, in
the mucous flakes and liquid contents of the intestinal canal of
cases of Asiatic cholera. It is stated that they were also detected
ASIATIC CHOLERA. 365
in a tank which contained the water supply of a neighbourhood
where cholera cases occurred; but comma-shaped organisms are
frequently present in sewage-contaminated water. Koch’s comma-
bacilli are aerobic, and their development is arrested by deprivation
of oxygen. They are destroyed by drying on a cover glass, but
retain their vitality longer when dried on silk threads. Cultures
are sterilised by exposure for fifteen minutes to 55° C., and by
various antiseptic substances.
|)
'
ii
Hi i),
ah Win
Fic. 154.—Pure-cuntivations In Nurrient GELATINY. «a, Kocu’s CHOLERA
Baciiius, twenty-four hours old. 0b, Finkier’s Bacrius, twenty-four
hours old.
Meruops or SrarniInG THE CoMMA-BACILLI or Kocu.
In cover-glass preparations they may be well stained in the ordinary
way, with anaqueous solution of methyl-violet or fuchsine, or by the
vapid method, without passing through the flame (p. 85, Babés’ method).
Nicati and Reitsch’s method.
A small quantity of the stools, or of the scraping of the intestinal
mucous membrane, is spread out on a glass slide and dried, then steeped
during some seconds in sublimate solution, or in osmic acid (1 to 100).
It is then stained by immersion in fuchsine-aniline solution (1 or 2
grammes of Bale fuchsine dissolved in a saturated aqueous solution of
aniline), washed, dried, and mounted in Canada balsam.
366 INFECTIVE DISEASES.
In sections of the intestine their presence may be demonstrated by :—
(a) Koch’s method.
Sections of the intestine, which must be well hardened in absolute
alcohol, are left for twenty-four hours in a strong, watery solution of
methylene-blue, or for a shorter time if the solution is warmed; then
treated in the usual way.
(b) Babes’ method.
Sections, preferably from a recent case of cholera, and made as soon as
possible after death, are left for twenty-four hours in an aqueous solution
of fuchsine, then washed in distilled water, faintly acidulated with acetic
acid, or in sublimate solution 1 in 1000, passed rapidly through alcohol,
and finally treated in the usual way.
Klein investigated cholera in India, and does not accept Koch’s
conclusions. With regard to the inoculation experiments, Klein
believes that the living choleraic comma-bacilli, even if introduced
in large numbers into the small intestine, are quite innocuous,
but capable of great multiplication if the intestine is previously,
from some cause or another, diseased; the chemical products of the
comma-bacilli then act as poisons analogous to the ptomaines
obtained from other putrefactive bacteria. The observations made
by Roy, Brown, and Sherrington, in Spain, tended to confirm
Koch’s views. Comma-bacilli were found to be present, in some
cases, in enormous numbers, and the frequency of their occurrence
led these observers to believe that they must bear some relation to
the disease. At the same time,
a as they failed to find them in all
cs Wee te \ cases, they regarded the existence
~ a C ; % ) Z “ of a causal relation as not proven.
) ) f /y They failed to find the Naples
~ a yA %, bacterium, or a small, straight
wo bacillus noted by Klein; and
they drew attention to certain
Fic. 155.—CoMMA-SHAPED ORGANISMS peculiar mycelium-like threads
WITH OTHER BacTERIA IN SEWAGE- .
CONTAMINATED WATER, x 1200. in the mucous membrane of the
: intestines ; but these cannot be
considered to have any significance. Methylene-blue has been
employed by Koch and others, including the author, for staining
sections of the intestine from cholera cases, and had they been
constantly present, it is hardly possible that such striking objects
could have been overlooked. Again, we must bear in mind that
hypho-mycetous fungi occasionally have been found to occur sapro-
phytically in the intestinal canal, as well as in the lungs, external
auditory meatus, and elsewhere. Cunningham, of Calcutta, maintains
ASIATIC CHOLERA.
367
that Koch’s comma-bacilli are not constantly found; and that
the comma-bacilli obtained from typical cholera cases show a
great variation in cultivation, and cannot be distinguished from
comma-bacilli from other sources.
Cunningham asserts that
comma-bacilli resembling Koch’s
are found in the intestine in
health. Sternberg, on the other
hand, made a number of examina-
tions of the evacuations of yellow
fever patients and healthy indi-
viduals, and failed to find any
¥Fia.156.—ComMaA-BACILLI OF THE MOUTH,
x 700 (Van ERMENGEM).
micro-organism resembling the cholera spirillum.
Various comma-bacilli have been isolated from different sources
and compared with Koch’s comma-bacillus. Comma-bacilli have been
Ny
Wey
= NaN
US;
Noyiira u
Lnr 2 AAS
=) LOvVe
WD Ute
») ASHES ON
= ss
nN) a Dic bs alll
Ls Ne
oo
we
Fic. 157.—FinKLER’s CoOMMA-BACILLI 5
FROM CHOLERA NOSTRAS, Xx 700
(Fiueer).
found in the mouth by Lewis ;
in cholera nostras by Finkler and
Prior; in cheese by Deneke ;
in hay infusion and sewage by
Weibel ; in the intestines of fowls
by Gamaleia, and in water by
Sanarelli,
Whether the comma-bacillus
is the cause of cholera or
not, its detection is an aid in
diagnosis. If we are dealing
with a case alleged to be one of Asiatic cholera, and a micro-
organism is found in the intestinal evacuations, which can be
differentiated from the comma-bacillus described by Finkler in
cholera nostras, and identified
with the comma-bacillus de-
scribed by Koch, we are justified
in regarding the case as one of
Asiatic cholera. But we cannot
diagnose Koch’s comma-bacillus,
with certainty, unless we know
the source of the culture. The
clinical symptoms of cholera in
man, and especially the presence
Fic. 158.—DENEKE’s COMMA-BACILLI,
yrom CHEESE, x 700 (FLiece).
of rice-water stools, must be taken into account, together with
the biological, morphological, and chemical characteristics of
the bacilli which are found to be present.
There are several
368 INFECTIVE DISEASES.
chemical tests which can be applied to cultures. According to
Frinkel, the Bujwid-Dunham test can be relied upon to distin-
guish Koch’s comma-bacillus from the comma-bacillus of Finkler-
Prior (cholera nostras), and from those found by Gamaleia. The
comma-bacilli are inoculated in broth containing peptone, and,
after twelve hours in the incubator, a drop of strong sulphurie
acid added to the culture will produce a red colour, owing to the
presence of indol. A test which distinguishes Koch’s comma-
bacillus from Finkler-Prior’s and Deneke’s was introduced by Cahen.
A solution of litmus is added to the broth, and the culture placed
in the incubator, until the following day; in the case of Koch’s
commas, the colour will have disappeared.
Koch points out that in the bacteriological. diagnosis of cholera
the first step is to examine the mucus in the evacuations, or in the
intestine if the examination is made after death. Cover-glass
preparations should be stained with dilute Ziehl-Neelsen solution.
Cultures are next made in peptone, and in eight hours will give the
indol reaction. In twenty-four hours the colonies may be examined
on plate-cultivations. The peptone cultures are prepared by adding
a trace of the choleraic evacuations, or of mucus containing the
bacilli, to a sterilised 1 per cent. solution of peptone, with ‘5 to 1
per cent. of common salt. The solution must be alkaline, and the
culture is placed in the incubator at 37° C. The pathogenic effects
can be ascertained by diffusing the bacilli from an agar-culture in
broth, and injecting it into the peritoneal cavity.
Toxic Products.—Brieger isolated several toxic products which he
had found in association with putrefaction, such as cadaverin and
putrescin ; but there were also present two new toxic substances, one
producing cramps and muscular tremors in inoculated animals, and
the other lowering the temperature and depressing the action of
the heart. Later, Brieger in conjunction with Frankel, succeeded
in isolating a tox-albumin from pure cultures. Guinea-pigs were
killed in two or three days, but rabbits had an immunity. Pfeiffer
found that cultures contained a poisonous principle which proved
fatal to guinea-pigs in extremely minute doses. It is broken up
by aleohol and by boiling, and secondary products formed, of very
much mitigated virulence. Similar toxic products were obtained
from cultures of both Finkler-Prior’s and Metchnikoff’s commas.
Protective Inoculation.—Haffkine has introduced a system of
protective inoculation, which is founded on the principle of inducing
the formation of antitoxins, or defensive proteids. Comma-bacilli
when first cultivated from a cholera patient are not sufficiently
ASIATIC CHOLERA. 369
virulent, and the virulence is increased by cultivation in the
peritoneal cavities of a succession of guinea-pigs. This successive
cultivation is carried on until a virus is obtained which proves fatal
in a few hours when inoculated into the peritoneum. A culture
from the peritoneum is obtained on an agar plate-cultivation, and
a pure sub-culture on agar is thoroughly shaken up with broth.
This constitutes the vaccinating fluid. It may be used as a living
vaccine, or the comma-bacilli killed by the addition of carbolic
acid.
Haffkine, having studied the pathological and physiological effects
on some sixty persons, mostly scientists interested in the subject, and
finding the treatment to be harmless, transferred his operations to
localities in India affected by cholera. The inhabitants of the
northern part of India were the first to come forward and submit
themselves to the inoculation. In the course of the first year
22,703 were inoculated in the North-West Provinces and Oudh,
and in the Punjab. All classes of, the population were included.
In the second year operations were carried out in those parts of
the country where cholera always prevails, and where, therefore, the
method could be more satisfactorily tested.
From March 1894, to July 1895, 19,473 individuals were
inoculated in some of the most affected localities.
From observations made at Calcutta by Dr. Simpson, from March
1894 to August 1895, cholera occurred in 36 houses containing
inoculated people. There were 521 inhabitants in the infected
houses, of whom 181 were inoculated from 1 to 459 days before the
occurrence, while 340 remained uninoculated. The uninoculated had
45 cases with 39 deaths from cholera; the inoculated had 4 deaths,
1 occurring 451 days after the first inoculation, and 3 others from
1 to 4 days after the first inoculation. These four cases had not
been re-inoculated. If the occurrences in inoculated and non-inocu-
lated during the first 10 days were set aside, and those considered
that occurred after the 10 days expired, then, according to Dr.
Simpson, the proportion of cases was 19:27 and that of deaths 17:24
times smaller in the inoculated then in the uninoculated.
Cholera broke out in the Gya gaol, and inoculations were made
after 6 cases, with 5 deaths, had occurred. During the stay of the
prisoners in the gaol, there were 209 uninoculated, with 7 cases and
5 deaths, and 211 inoculated, with 5 cases and 4 deaths.
In July and August in the same year cholera attacked the East
Lancashire Regiment. Out of 773 men there were 133 inoculated
and 640 uninoculated,
24
370 INFECTIVE DISEASES.
The occurrences of cases and deaths were :—
In 640 uninoculated 120 cases (18°75 7%), 79 deaths (12°34 %).
In 133 inoculated 18 cases (13°53 %), 13 deaths (9°77 7).
These results were, it is said, due to the weakness of the vaccines
procurable at that period of the work, and to the small doses used.
There were a great many records kept of the results of inocula-
tion of coolies on tea estates in different localities. After a summary
of the results Haffkine concludes, in his Report to the Government
of India, that, in his opinion, the experimental stage was not yet
in so advanced a condition as to be completely closed; but that
the observations made and records collected justified steps being
taken to give the inoculations a more
: renee i nt extended trial.
a ci r in (
ul
\ |
CHOLERA NOostTRAs.
Cholera nostras, English cholera, or
English dysentery, produces an inflamma-
tion of the mucous membrane of the
bowels with croupous exudation. The
large intestine is commonly affected, and
| i
i}
the mucous membrane may be covered
with small superficial ulcers. The disease
is associated with severe diarrhea.
Finkler and Prior obtained a comma-
bacillus from the evacuations, which they
believed to be identical with the comma-
bacillus found by Koch in Asiatic cholera.
Koch pointed out that there were marked
differences in the biological character of
the two micro-organisms.
Spirillum Finkler-Prior (Comma-
:
:
I
Hh
iil
ry)
iy
i Hi
: (a
Fic. 159.—PursE-cuLtTivaTion
7
OF THE SPIRILLUM FINKLER-
Prior, In Nutrient Gera-
TINE. In thirty-six hours.
bacillus in Cholera nostras).—Curved rods,
thicker than the comma-bacillus of Koch,
and spirilla. The colonies on plate-
cultivations are very much larger than
those of the comma-bacillus of Koch of the same age. They have a
very faint yellowish-brown tinge, a well-defined border, and a distinctly
granular appearance. They liquefy nutrient gelatine very rapidly,
so that the first plate of a series is, as a rule, completely liquefied on
the day following inoculation, and the second plate in two or three
days more. In a test-tube cultivation in nutrient gelatine the
appearances are especially characteristic: the gelatine is very rapidly
CHOLERAIC DIARRHGiA FROM MEAT POISONING. 371
liquefied along the whole track of the needle, so that the cultivation
resembles a conical sack, or the finger of a glove turned inside out.
On a sloping surface of nutrient agar-agar a white moist layer forms
very quickly, On potato they grow at the ordinary temperature
of the air, producing a brownish layer and corrosion of the surface
of the potato. They have been shown to be pathogenic.
CHoLtERAIc Diarrua@a From Mzat Poisontna.
There are two varieties of choleraic diarrhea from meat poisoning,
and both are associated with vomiting, diarrhea, pain in the abdo-
men, in severe cases followed by suppression of urine, collapse, and
death. These conditions are brought about by poisonous foods, and
include those cases of poisoning by tinned meats, pork pies, hams,
cheese, sardines, and other articles of food improperly prepared. In
most cases putrefaction has taken place, owing to the action of
various bacteria. Associated with their growth we find highly
poisonous substances, but no bacteria are found in the body in these:
cases. They are all due to chemical poisoning ; but Klein has also
described cases of poisoning due to the growth of bacteria without
the presence of putrefaction. The latter were of the nature of an
infectious disease. In the Welbeck. poisoning cases, described by
Ballard, the poisonous hams contained a short bacillus, which was
also found in the kidney and spleen in the fatal cases in man. In
the Carlisle epidemic, which was due to poisonous pork pies, the pork
and gravy stock proved fatal to mice, and from the infected mice
a bacillus was cultivated, which, administered to mice by feeding or
subcutaneous inoculation, produced enteritis, diarrhcea, and congestion
of the lungs.
Gartner cultivated Bacillus enteritidis from the spleen in a
fatal case of meat poisoning. Gaffky obtained a similar bacillus in
cases of gastro-enteritis, following the consumption of meat and
sausages, which had been made of horseflesh.
Bacillus of Choleraic Diarrhoea from Meat-poisoning
(Klein).—Rods from 3 to 9 win length, 1°3 » wide, rounded at their
extremities, singly or in chains of two. Spore-formation occurs,
the spores being 1 m thick, oval, and situated in the centre or at
the end of the rod.
Feeding mice with the bacilli and inoculation produced positive
results. At the autopsy, pneumonia, peritonitis, pleuritis, enlargement
of the liver and spleen, and hemorrhages were observed, and bacilli
were present in the blood and exudations of these animals, They
372 INFECTIVE DISEASES.
occurred in the blood and juices, and especially in the glomeruli of the
kidneys, of several fatal cases of choleraic diarrhea.
Bacillus enteritidis (Girtner).—Short rods in pairs, and short
chains. They are motile; spore-formation not observed. Colonies
are granular, and old colonies at the margin have an appearance of
Fic. 160.—TropicaL DysEnTERY. Mucous membrane of large intestine some
months after an acute attack: a,a, representing remains of mucosa ; b,b, inter-
vening parts corresponding to the muscularis (HAMILTON).
powdered glass. On the surface of gelatine a thick greyish-white
film develops, which in time becomes wrinkled. In the depth of
gelatine a white filament forms. The gelatine is not liquefied. On
agar the film is slightly yellowish. On potato it is similar in colour,
moist and shining. On blood serum it is very similar, Mice fed
CHOLERAIC DIARRHEA IN FOWLS. 373
with the bacilli die in one or two days. Subcutaneous injection is
fatal in guinea-pigs and rabbits in from two to five days. Dogs,
cats, and fowls are immune.
The bacilli were obtained from a cow suffering from a disease
associated with diarrhoea, and from the spleen of a man who died
twelve hours after partaking of the flesh of this animal.
DysEnTEry.
Dysentery is a disease of tropical climates associated with in-
flammation and ulceration of the large intestine (Fig. 160). At first
the discharge from the bowel is a whitish or brownish mucus,
which soon becomes blood-stained; later the evacuations become
thin and watery, with altered blood clots, fragments of mucous
membrane, and pieces of false membrane ; and in some cases they
become purulent. The virus is believed to be in the intestinal
discharges, which by contaminating water or soil may give rise to
other cases,
Micrococci have been found in dysentery, but the micro-organism
which has received most attention is a protozoon, the Ameba coli,
which will be described in another chapter.
Cuoteratic Diarruaa In Fow1s.
Choleraic diarrhea in fowls, or gastro-enteritis cholerica, is an
infectious disease of fowls, occurring in Russia during the summer.
The disease is very like fowl-cholera. The birds are sleepy, and
suffer from diarrhea, but the temperature is not raised, as in
fowl-cholera. After death there is usually an abundance of greyish
liquid in the small intestine, which is stained with blood. It was
investigated by Gamaleia, who found a comma-bacillus, to which he
gives the name Vibrio Metchnikovi.
Spirillum of Fowl-enteritis (Vibrio Metchnikovi).—Curved
rods and spirilla; thicker, shorter, and more curved than Koch’s
commas. They are motile, and possess a single flagellum at one end.
They stain with the usual dyes. Spore-formation doubtful. In plate-
cultivations minute white colonies appear in from twelve to sixteen
hours, and the gelatine is liquefied. The colonies in about three
days resemble those of both Finkler-Prior’s and Koch’s comma-bacilli,
some colonies being more like the one kind, and some like the other.
In the depth of gelatine the growth is very much like that of Koch’s
comma-bacillus, possessing the characteristic air-bubble appearance.
374 INFECTIVE DISEASES.
On agar a slightly yellowish growth is obtained, resembling that
of Koch’s commas ; on potato a yellowish-brown or chocolate layer
develops after incubation at the temperature of the blood, very
similar to cultures from Asiatic cholera. Broth becomes turbid, and
a wrinkled film forms on the surface; the addition of sulphuric acid
gives the indol test. The spirilla grow in milk, and coagulate it;
the milk becoming strongly acid, and the casein being precipitated.
They are pathogenic in chickens, pigeons, and guinea-pigs. Pigeons
die in about twelve hours after a subcutaneous injection ; and the
spirilla are found abundantly in blood from the heart. Guinea-pigs
die from acute septicemia in about twenty-four hours. The spirilla
are found in the blood and internal organs. Inoculation of pigeons
and guinea-pigs with sterilised cultures will produce immunity.
CHAPTER XXVIII.
TUBERCULOSIS.
TUBERCULOSIS is a communicable disease of man and animals, charac-
terised by the formation of new growths associated with the presence
of the tubercle bacillus. Von Bayle, in 1810, was the first to describe
little growths like millet seeds, which were considered to be character-
istic of consumption or phthisis. Laennec, in 1834, attached much
more importance to the existence of caseous matter and classified
miliary tubercle, crude tubercle, granular tubercle, and encysted
tubercle, as varieties of tuberculosis. Virchow would not accept
all these varieties as tubercular, and only regarded those conditions
associated with the presence of miliary tubercles as genuinely
tubercular. Laennec’s so-called crude tubercle, for example, was
simply due to pneumonic caseation. Villemin threw entirely fresh
light upon this controversy by proving that tuberculosis was a
communicable disease. Rabbits and guinea-pigs, inoculated with
tubercular sputum or caseous tubercle, developed miliary tubercle in.
a few weeks. Sanderson confirmed these experiments, and pointed
out that foreign bodies would produce experimental tuberculosis
in rabbits. Cohnheim also confirmed the experiments of Villemin,
and maintained that tuberculosis was a specific inoculable disease,
and, therefore, everything was tubercular which, on inoculation,
produced tuberculosis. Koch, in 1882, announced the discovery of
the tubercle bacillus, and expressed the opinion that without the
tubercle bacillus there could be no tuberculosis, Tubercle was
defined as tissue containing the tubercle bacillus, whatever might
be the clinical manifestations of the case, or the microscopical and
naked-eye appearances of the diseased parts,
A tubercle is a small growth about the size of a millet seed. In
the early stage it is circular, hard, grey in colour, and lustrous; but
when it undergoes necrosis and caseation it becomes soft and yellowish.
‘Tn the very early stage it consists of a little collection of round
375
376 INFECTIVE DISEASES.
cells, in which it is possible, though often with extreme difficulty,
to demonstrate the tubercle bacillus. The cells originate in the
proliferation of endothelial connective tissue and white blood cells.
Later on, large oval or circular multi-nucleated cells, or giant cells,
make their appearance. The tubercle bacilli are only occasionally
found in the interior of human giant cells, whereas in the lower
animals, in equine and bovine tuberculosis more especially, the bacilli
are often present in great numbers, and very commonly in the form
of conspicuous rings, visible under a low power of the microscope.
Hic. 161.—TuBERcLE oF THE LUNG IN A VERY EARLY STAGE, x 400: a, An alveolar
wall; 6, blood-corpuscles in capillaries of the same; c, blood-corpuscles
extravasated into the alveolar cavities; d, alveolar capillaries filled with
blood-corpuscles carried forward by the tubercle which is growing into the
alveolar cavity ; ¢, large endothelium-like cells, of which the tubercle in this
stage is mainly composed ; f, portion of a branch of the pulmonary artery
injected (HamILtTon).
Whether the absence of blood-vessels or the action of the bacillus is
the main factor in producing caseation, is an open question. When
suppuration follows caseation, as commonly happens in tuberculosis
of the lungs in man, and in experimental tuberculosis in animals, an
wbscess forms. In cattle there is a remarkable tendency to the
formation of calcareous deposit in the caseous masses.
The tubercle may not degenerate and die, but live and develop.
‘TUBERCULOSIS. 377
The giant cells, which are more or less central, have been described
as sending off processes, which, by dividing and subdividing, and
Fic. 162.—Primary TupercLe or LuNG TWO TO THREE WEEKS OLD, x 50:
a, Portion of wall of a branch of the pulmonary artery ; b,b, giant cells with
concentric arrangement of fibrous tissue; c, centre of tubercle beginning to
caseate ; d, small branch of pulmonary artery seen on transverse section ;
¢, injected capillaries of the alveolar walls (HAMILTON).
interlacing, form a reticulum, or
meshwork. Towards the periphery
of the tubercle the reticulum may
become arranged in the form of a
capsule as the age of the tubercle
advances, and the reticular giant
cell becomes eventually converted
into fibrous tissue. The bacillus has
disappeared, and the tubercle has
healed.
Giant cells cannot be relied upon —
to indicate tuberculosis. They are
not always present in tubercu-
losis, and they are not peculiar to
tubercle, being found, for example,
in actinomycosis. The only certain
indication of tuberculosis is the pre-
sence of the tubercle bacillus, which
Fic. 163.—Larce Ova Giant CELL
yRoM TUBERCLE oF Lune x 300:
a, Granular centre; b, nucleated
periphery forming a mantle-like
sheath; c, processes from the
same.
378 INFECTIVE DISEASES.
can be revealed either by microscopical examination of the suspected
tissue, or after inoculation in guinea-pigs.
Bacillus Tuberculosis (Koch).—Rods, 2 to 4 » and occa-
sionally 8 » long, very thin, and rounded at the ends. They are
straight or curved, and frequently beaded, and occur singly, in pairs,
or in bundles; there are algo involution forms and short branched
threads. Spore-formation is observed in old cultures. They are
non-motile. In the interior of giant cells they are often accompanied
by grains which exhibit the same colour reaction.
The bacilli in tissue sections of bovine tuberculosis are shorter
and less granular than those in human tubercular sputum, but in
milk they are quite as long, and even longer, and very distinctly
granular or beaded, and are thus brought much closer, morpho-
logically, to the bacilli in human sputum. Speaking generally,
however, the average length of the human bacilli is greater than
the average length of the bacilli in cow’s milk, but the longest of
the bovine bacilli cannot be distinguished in length from the longest
human bacilli. There are, however, exceptional cases, for in some
preparations of pus from human lungs the bacilli are remarkable,
not only for their thinness, and their uniformly beaded character,
but more particularly for their extraordinary length. They should
be compared with other preparations, in which the bacilli, though in
human sputum, are sometimes much more distinctly rod-shaped,
much shorter and thicker, with complete absence of any beaded
appearance. Neither length nor granularity is a characteristic
sufficient to denote any specific difference between human and bovine
bacilli. The author has examined minutely the bacilli in tuberculosis
of other animals, such as the horse, pig, and cat; and of birds—the
fowl, guinea-fowl, pheasant, and ostrich. Here, again, minute
morphological differences can be observed. For example, in many
cases in fowls the bacilli are conspicuously short and straight. In
the liver and lungs of an ostrich, packets of short rod-forms are
found, while in other parts of the same sections the bacilli attain
a very great length. Many of the long, sinuous forms exhibit a
peculiar terminal enlargement. There are also short rods with a
similar appearance, and free ovoid bodies, singly and in groups, which,
from their connection with the bacilli, and their sharply defined
outline in the free state, are similar to spores in old cultures.
Thus, morphological differences are found under different circum-
stances, and within limits the morphology of the tubercle bacillus
varies with its environment.
Koch first succeeded in cultivating the bacillus by employing
DESCRIPTION OF PLATE XI.
Bacillus tuberculosis.
The figures in this plate represent- the bacilli of tuberculosis in
different animals, examined under the same conditions of amplifica-
tion and illumination. x 1200. Lamp-light illumination,
Fig. 1.—Bacilli in pus from the wall of a human tubercular cavity. In
this specimen the bacilli are shorter than those in tubercular sputum,
and are very markedly beaded.
Fig. 2.—Bacilli in pus from a tubercular cavity from another case in man.
They are present in the preparation in enormous numbers, The proto-
plasm occupies almost the whole of the sheath, and the bacilli are
strikingly thin and long.
Fig. 3.—Bacilli in sputum from an civaned case of phthisis, showing
the ordinary appearance of bacilli in sputum; some beaded, others
stained in their entirety; occurring both singly and in pairs, and
in groups resembling Chinese letters.
Fie. 4.—Bacilli in « section from the lung in a case of tuberculosis in man,
The bacilli in human tuberculosis are found in, and between, the tissue
cells ; and sometimes, as in equine and bovine tuberculosis, in the
interior of giant cells, but not so commonly.
Fic. 5.—From a cover-glass preparation of the deposit in a sample of milk
from a tubercular cow. The bacilli were longer than the average
length of bacilli in bovine tissue sections, and many were markedly
beaded.
Fia. 6.—From a section of the brain in a case of tubercular meningitis in a
calf, showing a giant cell containing bacilli with the characters usually
found in sections of bovine tuberculosis.
Fig. 7.—From a section of the liver of a pig with tubercle bacilli at the
margin of a caseous nodule.
Fig. 8.—From a cover-glass preparation of a crushed caseous mesenteric
gland from a rabbit infected by ingestion of milk from a cow with
tuberculosis of the udder,
Fig. 9.—From a section of lung in a case of equine tuberculosis, showing a
giant cell crowded with tubercle bacilli.
Fig. 10.—From a section of lung from a case of tuberculosis in the cat, with
very numerous tubercle bacilli.
Fic. 11.—From a cover-glass preparation of a crushed caseous nodule from
the liver of a fowl, with masses of bacilli. These are for the most part
short, straight rods; but other forms, varying from long rods to mere
granules, are also found.
Fig. 12.—From sections of the liver and of the lung in a case of tubercu-
losis of a Rhea, Isolated bacilli are found, as well as bacilli packed in
large cells, colonies of sinuous bacilli, and very long forms with terminal
spore-like bodies and free oval grains.
The preparations from which these figures were drawn were all
stained by the Ziehl-Neelsen method, with the exception of the
first, which was stained by Ehrlich’s method.
Plate XI.
BACILLUS ee a SIs.
| EMCrockeshanle fect. \
Vincent Brooks,Day & Son, lath.
\
TUBERCULOSIS. 379
blood serum. Solid blood serum, with or without the addition of
gelatine, was employed, and the cultures incubated at 37°C. The
growth takes place very slowly, and only between the temperatures
of 30° C.and 41° C. In about eight or ten days the growth appears
as little whitish or yellowish scales and grains.
The bacillus can also be cultivated in a glass capsule, on blood
serum, and the appearances of the growth studied under the
microscope. The scales or pellicles were described by Koch as made
up of colonies of a perfectly characteristic appearance, which may
be still further studied by making a cover-glass impression. They
are then seen to be composed of bacilli, arranged more or less
with their long axis corresponding with that of the colony itself,
and with an appreciable interval between the individual bacilli.
The colonies themselves appear as fine curved lines, the smallest
being mostly S-shaped. Longer colonies have serpentine twistings
and bendings, which often
recall the curves of fancy
lettering. The ends of the ~ ‘N
lines run to sharp points, 4 oN
but the middle of the
growth is spindle-formed. ewe
The youngest colonies are =
extremely delicate and
narrow, but the older ‘
colonies increase in size,
are thicker across, and,
blending with each other,
gradually obliterate the
characteristic appearances; a lamellated growth results, which
increases, and gives the appearance to the naked eye of the scale
or pellicle already described. The blood serum is not liquefied
unless putrefactive bacteria contaminate the culture. A fresh tube
can be inoculated with one of the little scales, and a new generation
started. The scales gradually increase in size, and consist entirely
of bacilli. In about three to four weeks the cultivation ceases to
increase, and it is then necessary to inoculate a fresh tube.
In liquid blood serum a film forms on the surface of the liquid,
and is easily broken by agitation. In neutralised broth there is
very little indication of success. When a triturated culture is added
to the broth, a granular, sandy, whitish deposit collects’ at the
bottom of the vessel, with indications of an increase in amount.
Koch also tried nutrient agar-agar, which did not prove to be at
Fic. 164.— Bacintus TUBERCULOSIS, FROM
TUBERCULAR Sputum, x 2500. From Photo-
graphs.
380 INFECTIVE DISEASES,
BACILLUS TUBERCULOSIS.
a c
Fic. 165.—PUuRE-CULTIVATIONS ON GLYCERINE-AGAR FROM HuMAN TUBERCULAR
Spurum. a, After six months’ growth. (Fifth sub-culture.) b and c, After
ten months’ growth. (Fourth sub-cultures. )
TUBERCULOSIS.
381
all a favourable medium. Some increase took place, but there
was no continuous growth over the inoculated area.
Glycerine Agar-agar.—Nocard and Roux were among those who
worked at the subject and confirmed Koch’s
observations. Nocard attempted to get
cultures of avian tuberculosis on blood serum
to which peptone, salt, and cane sugar had
been added. The results were more success-
ful than with ordinary serum. But he
encountered a difficulty in the rapid drying
of the surface of the medium, which rendered
the tubes unfit for use. It occurred to
Nocard and Roux to obviate this by the
addition of a hygroscopic agent, and accord-
ingly they added sterilised glycerine. The
result, which far exceeded their expectation,
evidently was not solely attributable to the
prevention of
Fic.
167.—PURE-CULTI-
VATION IN GLYCERINE
AGAR-AGAR,—A SUB-
CULTURE FROM A PURE-
CULTURE IN GLYCERINE-
MILK.
In two months.
desiccation. Following up
their discovery, and
being anxious to find he. 1G3—euaaeonme
a medium more easily VATION IN GLYCERINE
prepared than blood JAGUTEAA after ten
months’ growth.
serum, they added
6 to 8 per cent. of glycerine to ordinary
nutrient agar-agar. The bacillus grew so
abundantly in this mixture that a culture
in fifteen days equalled in extent a culture
on blood serum which was several weeks old.
The bacillus was found to grow abundantly
in veal broth, to which glycerine had been
added in the proportion of 5 per cent., the
bottom of the flask being covered in about
three weeks with a flocculent deposit, having
some resemblance to anthrax cultivations
in liquid media. In beef broth, chicken
broth, and in Cohn’s liquid, cultures were
obtained after the addition of glycerine.
Description of Cultivations on Glycerine
Agar-agar.—the cultivations on the sloping
surface of obliquely solidified glycerine agar-
agar begin to appear in from four toysix days as very minute white
colonies, These steadily increase in size, and either look moist and
382 INFECTIVE DISEASES.
smooth, or, even at this early stage, appear dry and crinkled.
According to the number of bacilli inoculated, the colonies will
either remain isolated or coalesce and form a more or less continuous
film. If the nutrient agar-agar has only recently been prepared,
there is usually a quantity of liquid present, and the bacillus forms
a white coating over the inoculated area and beyond it. The
appearances are much more characteristic when this medium is,
comparatively speaking, dry. A semi-transparent membranous
growth develops, thickens, and assumes a characteristic lichenous
appearance. Such a culture, examined with a pocket lens, resembles
a model in wax in miniature of the folds of the gastric mucous
‘ membrane. In about six weeks to two months the culture has fully
developed. In old cultures, especially when the individual colonies
remain isolated, the appearance is very characteristic. Some cul-
tures in appearance closely resemble cultivations on blood serum,
The consistency of the growth depends upon the character of the
soil and the age of the culture. When the medium is moist the
growth is moist and viscous, but more often it is distinctly tallowy,
and in old and dry cultures scaly and friable.
Cultivations in Glycerine Broth.—In a few days minute flakes are
visible, which steadily increase in size, and subside to the bottom of
the flask, forming in time a very copious deposit. On shaking the
flask, this deposit, which is extremely tenacious, rises in stringy
masses, and gives an appearance which is more or less character-
istic. If the flask is left undisturbed, a delicate veil-like film forms
over the surface, which can be readily broken up by gentle agitation,
forming flakes which gradually sink in the liquid. If undisturbed
for several weeks this film increases in thickness, is irregularly
fissured, and has more the appearance of masses of tallow floating
on the surface. The growth also may be seen to extend up the side “_
of the flask above the liquid. Pasteur or Erlenmeyer flasks can be
employed for these cultures. Solidified egg-albumin added to the
glycerine broth seems to increase the amount of growth, which
clings to the albumin, and waves to and fro in the liquid when the
flask is gently shaken. The author has confirmed the observation
of Nocard and Roux, that sub-cultures from glycerine agar-agar, or
from glycerine broth, will give cultures in ordinary broth without
glycerine. Ordinary broth with egg-albumin, and without glycerine,
will also give a good growth when inoculated from previous sub-
cultures, although the attempt to produce primary cultures in these
media has hitherto failed.
Cultivations in Glycerine-Milk, and other Media.—In milk the
TUBERCULOSIS. 383
author found there was only a slight increase in the number of bacilli
inoculated, but milk with glycerine in the proportion of 5 per cent.
forms a more favourable medium. The author has also cultivated the
bacillus on sterilised urine and glycerine, and ordinary nutrient
gelatine with 5 per cent. of glycerine. On potato the growth of
the bacillus is extremely slow. Beevor succeeded in obtaining
cultures at the ordinary temperature of the room.
Examination of Cultivations.—To examine the bacilli in these
various preparations the author prefers to use Neelsen’s method,
floating the cover-glasses for from five to ten minutes on warm
carbolised fuchsine, and passing them through dilute sulphuric acid.
In some cultures the bacilli are shorter and thicker than is commonly
observed in human sputum, and they are for the most part without
the beaded appearance. In old cultures on glycerine agar-agar the
number of granular or beaded bacilli increases, and there are also
numerous peculiar forms, There are bacilli, sometimes two or
three times the length of an ordinary bacillus, provided with a
club-shaped enlargement at one or both extremities, and in rare
cases with lateral branches. They are no doubt identical with the
bacilli with swollen extremities and the branched forms observed
by Nocard and Roux.
In milk the appearance is very striking, many bacilli attaining
in old cultures a great length, and all are more uniformly beaded
than in any other cultivations. Staining preparations by the method
of Gram in all cases exaggerate this appearance.
The important part played by the environment is shown by the
morphological differences observed in artificial cultivation under
varying conditions, and by the fact that by successive cultivation
the bacillus can be educated to grow upon a medium which is un-
suitable for obtaining primary cultures.
Impression preparations of the growth of the bacillus on the
surface of glycerine agar-agar in capsules show a tendency to the
formation of serpentine colonies, composed of bundles of more or
less parallel bacilli.
Spore-formation.-_In old cultivations true spore-formation can
readily be observed, both in stained and unstained preparations. In
the latter case they are recognised in the form of one or two highly
refractive bodies in individual bacilli.
Toxic Products of Cultwres.—The poisonous substances found in
cultures, and the composition and use of tuberculin, have already
been described (p. 43).
Inoculation Experiments.— A relatively small portion of a culti-
384 INFECTIVE DISEASES.
vation inoculated into the subcutaneous tissue, into the peritoneal
or pleural cavities, into the anterior chamber of the eye, or directly
into the blood stream, produces after three or more weeks artificial
tuberculosis in guinea-pigs and rabbits. Dogs and cats can also be
infected by experimental inoculation.
When a trace of tubercular virus is inserted subcutaneously in
the thigh of a guinea-pig, in about a week or ten days a chain of
enlarged glands will be easily felt in the vicinity of the seat of
inoculation. This affords an unfailing test, which can be applied
when there is difficulty in ascertaining by the microscope the presence
of the bacilli in the material under examination. It also affords a
valuable method for testing the effects of antiseptics on tubercular
virus. The appearances observed at the autopsy are swollen
lymphatic glands, in the neighbourhood of the inoculation, followed
by softening and abscess ; enlargement of the spleen and liver, with
formation of caseous tubercles ; and tubercular deposits in the lungs,
bronchial glands, and peritoneum.
After inoculation of the eye, grey tubercles appear on the iris,
and undergo enlargement and caseation, followed by tuberculosis
of the eyeball and organs generally.
The bacilli appear to be the direct cause of tuberculosis, and
the presence of the bacillus in the sputum of patients is a distinctive
sign of the existence of this disease. The detection of the bacillus
has, consequently, become a test which is constantly applied.
The bacilli are found in all tubercular growths of man, monkeys,
cattle (Perlsucht), birds, and many other animals, and in cases of
artificial tuberculosis, in rabbits, guinea-pigs, cats, etc. In man the
bacillus can be detected in the tissues, in the sputum, in the blood,
and in the urine.
Tuberculosis may also be produced by inhalation and feeding
experiments. The channels of infection in man are also most
probably the pulmonary and intestinal mucous membranes. The
possibility of inoculation of skin wounds is open to doubt. The
bacilli or their spores are inhaled from the air, or taken in with
food. Morphologically identical bacilli have also been observed, but
very sparsely, in sections of lupus.
Meruops oF EXAMINING THE TUBERCLE BaciILuus.
Numerous methods have been recommended for examining the
tubercle bacillus. A few of these will be described, as many are
only of historical interest,
TUBERCULOSIS. 385
The Ziehl-Neelsen method is preferred by the author both for
sections and cover-glass preparations.
Koch's original method.—Cover-glass preparations or sections are laid
in Koch’s solution (No. 23, ¢) for twenty-four hours, or for one hour if
the solution is warmed to 40°C. Rinse in water ; immerse in a watery
solution of vesuvin for two minutes ; rinse again in water, and examine ;
or, after rinsing in water, treat with alcohol, clove-oil, and Canada
balsam.
Ehalich’s method.—Cover-glass preparations are allowed to float in a
watch-glass, containing a solution of gentian-violet or fuchsine, added to
aniline water. A saturated alcoholic solution of the dye is added till
precipitation commences (10 ce. aniline water, and 10 to 20 drops of the
colour solution). The cover-glasses are left in the solution for about
half an hour ; then washed for a few seconds in strong nitric acid (one
part commercial nitric acid to two of distilled water), and rinsed in
distilled water. After-stain with vesuvin or methylene-blue, rinse in
water, dry and preserve in Canada balsam.
Ehrlich-Koch method. °
Saturated alcoholic solution of methyl-violet or fuchsine 11
Aniline water : : j 100
Absolute alcohol . : ; 10
Preparations are left for twelve hours in this solution (colouring of
the cover-glass preparations can be expedited by warming the solution).
Treat the preparations with (1 to 3) solution of nitric acid a few
seconds.
Wash in alcohol (60 per cent.) for a few minutes (cover-glass prepara-
tions need only be rinsed a few times). After-stain with diluted solution
of vesuvin or methylene-blue for a few minutes.
Wash again in 60 per cent. alcohol, dehydrate in absolute alcohol.
Clear with cedar-oil, mount in Canada balsam.
Rindfleisch’s method.—Prepare a solution composed of
Saturated alcoholic solution of fuchsine : 10 drops
Aniline water . F : . 2 drams.
Pour it into a watch-glass, and float the cover-glass ; warm the watch-
glass over a spirit-lamp until steam rises. Remove it from the flame,
and set it aside for five minutes. Take out the cover-glass, and transfer
it for a few seconds to acidulated alcohol (two drops of nitric acid in a
watch-glass full of alcohol). Wash in distilled water, dry, and preserve
in balsam. After-stain, if necessary, with Bismarck-brown, or methylene-
blue.
Gibbcs’ method.— Cover-glass preparations are placed in Gibbes’ double-
staining solution which has been warmed in a test-tube, and, as soon as
steam rises, poured into a watch-glass. They are allowed to remain for
five minutes, and then are washed in methylated spirit till no more colour
comes away, dried in the air or over a spirit-lamp, and mounted i
Canada balsam, If the sulution is used without warming, the cover-glasses
25
386 INFECTIVE DISEASES.
must be left in it for an hour. Sections are treated on the same
principles, but must be left in the solution for several hours. The
crumpling of the sections by the action of nitric acid is avoided.
Baumgarten’s method.—Cover-glass preparations of sputum are made
as already described, and immersed in a very dilute solution of potash
(1 to 2 drops of a 33 per cent. solution of potash in a watch-glass of dis-
tilled water). The cover-glass is pressed down on a slide, and examined
with a high power. The bacilli can be thus examined in the unstained
condition, and to avoid any mistake from confusion with other species,
the cover-glass can be removed, dried, passed through the flame, and
stained with a drop of an aqueous solution of fuchsine, or gentian-violet.
The putrefactive bacteria are stained, but the tubercle bacilli remain
absolutely colourless.
Baumgarten’s new method.—A solution is prepared as follows: Drop
4 to 5 drops of concentrated alcoholic methyl-violet solution into a small
watch-glass full of water. (a) Stain the sections in this solution, wash
them in water, and decolorise in absolute alcohol (five to ten minutes) ;
or, before treating with alcohol, immerse the sections for five minutes in
a half-saturated solution of carbonate of potash. Pass through clove-oil,
and mount in a mixture of Canada balsam, free from chloroform, and
clove-oil (equal parts). The object of this process is to differentiate the
tubercle bacilli from chance bacteria, inasmuch as the tubercle bacilli
are gradually decolorised by the clove-oil. (0) Sections stained in the
above solution are placed for five minutes in alcohol, and then in a
concentrated solution of Bismarck-brown in 1 per cent. solution of acetic
acid. The after-treatment may be conducted as already described.
_ Ziehl-Neelsen method.—Cover-glass preparations may be quickly stained
in Neelsen’s solution warmed in a watch-glass till steam rises. Sections
are left for from five to ten minutes in the solution, and then washed in a
watery solution of sulphuric acid (25 per cent.), rinsed in distilled water,
and immersed in methylene-blue solution. After two or three minutes
they are passed through alcohol and oil of cloves, and mounted in Canada
balsam.
Frénkel’s method.—Sputum preparations are rapidly double-stained
by the following method: Prepare a solution by adding concentrated
alcoholic methyl-violet or fuchsine solution, drop by drop, till opalescence
arises, to 5 ccm. of aniline-water heated to 100° OC. Float the prepared
cover-glasses two minutes in the warmed solution. The process of after-
staining and decolorisation is effected by placing the preparation for one
to two minutes in one of the following solutions: for fuchsine-stained
preparations, a saturated solution of methylene-blue in a mixture of
Alcohol ; : 50
Distilled water : ; 30
Nitric acid : , : 20
which is filtered before use ; for preparations stained in methyl-violet, a
saturated solution of vesuvin may be used in
Alcohol , : 70
Nitric acid ‘ , : ; : ; 30
TUBERCULOSIS. 387
Ehrlich’s Method and Eosin.—The author has found that after sections
have been stained with methyl-violet and Bismarck-brown by Ehrlich’s
method, as described by Koch, they may with advantage be immersed in
a weak alcoholic solution of eosin, then rinsed in clean absolute alcohol,
clarified with clove-oil, and mounted in Canada balsam. The giant cells
are then stained pink, while their nuclei are brown, and the bacilli blue.
TUBERCULOSIS’ IN Man,
The disease manifests itself in various forms in man, and most
frequently in the lungs, producing phthisis or consumption. The
sputum contains the bacilli in large numbers, and is extremely
virulent. Scrofula and lupus are forms of tuberculosis; they are
Fic. 168.--SecTION THROUGH A LUPUS NODULE OF THE Noss.
probably produced by an attenuated variety of the tubercle bacillus.
Lupus can be distinguished from tuberculosis of the skin; and
scrofulous lymphatic glands are distinguished from tubercular glands
by the tendency of the latter to produce generalised tuberculosis.
This difference in the intensity of the virus in the two cases,
Lingard illustrated by the effect upon inoculated guinea-pigs.
Cavities in the lungs are often thickly lined with bacilli, They
are present in great numbers in the caseous matter, though in
equine and bovine tuberculosis this is not the case.
Whether the disease in man is contagious is an open question,
though numerous cases of supposed communication between husband
and wife, brothers and sisters, have been reported, and Ransome
388 INFECTIVE DISEASES.
showed that tubercle bacilli were present in the breath in phthisis,
On the other hand, the experience in consumption hospitals does not
Fic. 169.—Tupercutar ULceration ior Mucosa or Human Iteum.
Between the ulcers there are tubercular lymph-follicles (Hamiuton).
support this view, there being no evidence of the communication of
the disease to nurses and hospital attendants,
TUBERCULOSIS. 389
TUBERCULOSIS OF CATTLE.
In cattle the disease may occur as the result of inhaling bacilli,
or of ingestion with food. - It is very frequently found in the lungs;
and calves may be infected by milk from cows with tubercular
udders. Calves may also suffer from congenital tuberculosis, the
bacilli having been transmitted from the mother during gestation.
Breeding in-and-in, over-production of milk, and confinement with
insanitary surroundings, predispose to tuberculosis. The disease is
known in Germany as “ Perlsucht”; and in this country the lesions
on the pleura are known as
“grapes,” and the animals
themselves are commonly
called, “‘ wasters.”
The disease may also exist
in the lungs or in other
organs, in a limited form,
without any indication of
ill health. In such cases
the disease can be detected
by injection of tuberculin, a
marked rise of temperature
occurring in tubercular
animals.
In advanced cases, the
symptoms commonly observed Fc. 170. --SECTION OF Lurus OF THE SKIN,
A é x 700. Giantcell containing a tubercle
are cough, difficulty in breath-
bacillus (FLUGGE).
ing, staring coat, wasting, and
diarrhea; and if the udder is infected, nodules in the gland, and thin
bluish milk. In the lungs, after slaughter, a few small cheesy tubercles
may be found in animals apparently in perfect health and in prime
condition for the market. In advanced cases, the lungs on section
show large yellow masses, containing calcified matter, and the
bronchi may be full of yellowish pasty contents. The disease will be
found to involve the bronchial glands. The serous membraue may
be covered with little warts or grape-like masses. The lymphatic
glands may be enlarged to an enormous size. Tubercular ulceration
of the intestine is sometimes found, but not commonly. In tubercular
disease in the udder, a painless swelling is found which may affect
one or more quarters of the gland.
Transmission of Tuberculosis from Man to Cattle.—It is
390 INFECTIVE DISEASES.
for obvious reasons impossible to ascertain by experiment whether
tuberculosis can be transmitted from cows to man by milk or other-
wise; but some light may be thrown upon this important, question
by ascertaining the result of inoculating bovines with human tuber-
culosis. If calves can be infected with tuberculosis from a human
source by inoculation or ingestion experiments, and especially if
the effect of administering human and bovine tubercle to calves
by these means is found to be the same, such experiments will
not only serve to dispel any doubt there may be as to the identity
of the two affections, but they will strengthen the hands of those
Fic. 171.—Tusrrcu.osis or PLEURA; ‘‘GRAPE-DISEASE.”
who insist upon the necessity of more thorough inspection of dairy
cows, and of power to deal with tubercular animals.
Inoculation of a Calf with Human Tubercular Sputum.—The
author obtained sputum containing numerous bacilli from an
advanced case of phthisis. The sputum was shaken up with sterilised
salt solution and injected into the peritoneal cavity. A few weeks
afterwards the calf showed signs of illness. The animal looked dull,
did not feed well, had a slight cough, and showed less inclination to
move about than usual. These symptoms gradually increased, and
death occurred forty-two days after inoculation, Extensive lesions
TUBERCULOSIS. 391
were discovered at the post-mortem examination. The mesentery
was adherent to the abdominal wall, at the seat of the inoculation,
and to the rumen ; the liver was adherent to the diaphragm. There
was extensive tubercular deposit at the seat of inoculation, and
an abscess the size of a walnut. Extending over the mesentery
from this point there were hundreds of wartlike, fleshy, new growths,
some quite irregular in form, others spherical or button-shaped.
There were similar deposits on the under surface of the liver, on the
spleen, in the gastro-splenic omentum, and on the peritoneal surface
of the diaphragm. The spleen was adherent to the rumen, and
on dissecting away the adhesions another abscess was opened. The
lungs were congested and the pleure thickened. On microscopical
examination of sections extremely minute tubercles were found to
be disseminated throughout the whole of the substance of the lungs
_and liver, and tubercle bacilli were found in these and in the
peritoneal deposits. The abscesses contained Streptococcus pyogenes.
The calf died of pyemia, but sufficient time had elapsed for marked
local infection leading to generalised miliary tuberculosis.
TuBERCULOSIS IN RELATION TO THE PusLic Mink SupPty.
There is not the slightest doubt that when the udder is involved
the milk is highly virulent to the lower animals, and presumably
is, therefore, dangerous to man. The virulence of the milk was
first insisted upon by Klencke in 1846, and confirmed by Gerlach in
1869, and later, by others.
This subject was again brought forward with the discovery of
the tubercle bacillus, and the demonstration of its existence in the
milk in certain cases of bovine tuberculosis. Koch pointed out that
the milk only contained bacilli, and was only infective, when the
udder itself was tubercular. By this he explained the contradictory
results obtained by various experimenters with milk from cows un-
doubtedly suffering from “ Perlsucht.” Koch considered that positive
effects were obtained with milk when it happened to contain tubercle
bacilli, and negative with milk from which they were absent. Bang
in a number of cases verified the presence of tubercle bacilli in milk,
and, owing to the contradictory results of previous investigations,
repeated the ingestion experiments. The milk was found to be
virulent both to pigs and rabbits.
In this country Woodhead and M‘Fadyean tested milk for
tubercle bacilli. They examined six hundred cows in the Edinburgh
dairies, and fonnd thirty-seven suffering from mamumitis, but in only
392 INFECTIVE DISEASES.
six were they able to demonstrate the presence of tubercle bacilli in
the milk, and then only in small numbers.
Hirschberger found in twenty cases of tuberculosis in cattle
that the milk of eleven was virulent to guinea-pigs. Three cows out
of nine in which the disease was restricted to the lungs gave infected
milk. On the other hand, Nocard inoculated milk from eleven
tuberculous cows, of which only one had diseased udder, and only
this one gave infective milk. Bang injected rabbits with milk from
twenty-one cases of tuberculosis, with the udders apparently normal,
and the milk was virulent in two.
The author had two cases of udder tuberculosis under observation,
and as no experiments had at the time been made in this country
with milk known to contain tubercle bacilli, it was decided to study
the effect on rabbits, and test the results obtained by Bang. These
cases were both interesting and instructive, and may be referred to
in detail.
One was a case of advanced general tuberculosis. There was extreme
emaciation, general apathy, and a peculiar dull expression of countenance.
The skin was dry and harsh, the coat staring, and there was loss of hair
in patches about the face and neck. There was dulness on percussion
over a large area of the thorax, and the respirations were increased in
rapidity. There was also occasional cough and some diarrhoea. But the
most interesting condition was observed on examination of the udder.
The gland was swollen, especially posteriorly, and distinct induration
could be felt on examination. The deposit appeared to be more or less
limited to the posterior quarters. The cow evinced no pain during the
examination of the udder, not even on the application of firm pressure.
The author took samples in test-tubes of the milk from all four teats ;
when freshly drawn, it differed noticeably from the normal secretion. It
was a thin, watery, turbid fluid with whitish flakes in suspension, but it
was not gelatinous or muco-purulent in character, and was free from any
markedly yellow colour. After being set aside in the laboratory for
some hours it separated into a layer of cream and a turbid liquid of a
yellowish tint, while at the bottom of the test-tube there was a whitish
flocculent deposit, especially in the samples from the posterior quarters.
There were tubercle bacilli both in the cream and in the deposit. In
the cream they were only present in small numbers, and were detected,
therefore, only after careful search. But in the deposit they were readily
found, as in a cover-glass preparation there were sometimes four or five
in the field of the microscope.
The method adopted for the examination of this deposit was as
follows: The whole of the liquid in the test-tube was carefully poured
off, and a trace of the sediment spread out on a cover-glass. This was
allowed to dry, and passed through the flame, and stained in hot Ziehl-
Neelsen solution in the usual manner.
TUBERCULOSIS. 393
The other cow was also a case of general tuberculosis, and presented
somewhat similar lesions of the udder. The induration of the gland was
readily detected, and examination of the milk showed, as in the previous
case, the presence of tubercle bacilli.
It will be observed that in neither of these cases was the disease
limited to the udder ; in both the implication of the gland was part of
general tuberculosis.
Fre. 172.—Tusercutak ULCERATION OF THE INTESTINE OF a Cow.
The first cow was killed, and the following lesions were found at the
post-mortem examination.
Thoraw.—The lungs and bronchial glands were extensively invaded
with tubercular deposit. The glands were greatly enlarged and densely
fibrous, in many cases with central, stone-like masses, grating on section
against the edge of the knife. In the lung there was every stage, from
the early deposit to purulent cavities, cheesy masses, and calcified débris.
Abdomen.—There were a few caseous nodules in the liver, but none
in the spleen. The mesenteric glands formed an almost continuous chain
394 INFECTIVE DISEASES.
of large tumours, mostly with central cretification. Tubercular deposit
in the intestines could be recognised from the outside, and on laying
them open the mucous membrane was found to be studded with tuber-
cular ulcers. These ulcers were most numerous in the large intestine,
and varied in size from a sixpence to a florin. Some were circular, others
slightly irregular in form, and others again distinctly oval. In the latter
case they were generally situated with their long diameter transversely.
The base of the ulcer involved the muscular coat, and was irregularly
radiated. The margin was broad, and elevated above the general surface,
producing a ring-like appearance.
Mammary Gland.—The udder was infiltrated throughout with tuber-
cular new growth, but the invasion was most marked in the posterior
quarters. There was apparently very little tendency to caseation.
Microscopical Examination of the Udder.—In order to study the histo-
logical characters of the gland, and the distribution of the bacilli, sections
were stained with logwood and rubin, and others again with fuchsine
and methylene-blue, The tubercular new growth consisted of the usual
histological elements, round cells, epithelioid cells, and giant cells.
Healthy lobules here and there were sharply marked off from those in
which the growth was compressing and obliterating the alveoli in its
progress. Bacilli were present in the giant cells, and also distributed in
vast numbers throughout the tubercular tissue generally. Bacilli were
found in epithelioid cells close to the alveolus, and also between the cells
lining the alveoli, In parts also the new growth had involved the milk
ducts, and therefore it was easy to account for the presence of the bacilli
in the milk.
The bacilli were found in considerable numbers also in sections of the
intestinal ulcers,
EXPERIMENTAL INFECTION OF RABBITS.
Ingestion.—A rabbit received the contents of a test-tube which
had been filled with milk from one of the posterior teats, mixed with
a small quantity of bran. In four weeks there was commencing
emaciation ; later, diarrhoea set in, and death occurred exactly fifty-
eight days after administration of the milk. At the post-mortem
examination the mesenteric glands were found to be much enlarged
and caseous. A cover-glass preparation from a crushed gland
revealed numerous tubercle bacilli. On opening the intestines there
was a patch of ulceration, showing the point of access of the bacilli.
The intestinal ulceration was a reproduction, to a certain extent, of
the condition in the cow which had been the source of the virus.
Subcutaneous Injection.—A second rabbit was injected under the
skin of the back by means of a capillary pipette with about ten
drops of milk, including some of the deposit from the bottom of the
test-tube. The sample of milk had in this case also been taken from
DESCRIPTION OF PLATE XII.
Tubercular Mamumitis.
N
Fig. 1.—From a section of the udder of a milch cow. The tubercular deposit
is seen to invade the lobules of the gland. Lobules comparatively healthy
are marked off, more or less sharply, from the diseased ones in which the
new growth in its progress compresses and obliterates the alveoli. Stained
by the Ziehl-Neelsen method and with methylene-blue. x 50.
Fig. 2.—Part of the same preparation, On the right of the section part of a
healthy lobule is seen. On the left a lobule is invaded by tubercular new
growth composed of round cells, epithelioid cells and typical giant cells.
Tubercle bacilli can be seen both singly and collected in groups. They
are found in and between the cells, and in the interior of giant cells.
Bacilli may be seen between the cells lining an alveolus and projecting
into itslumen. x 800.
nt Brooks, Day & Som Lith
TUBERCULOSIS. 395
one of the posterior teats. The rabbit was placed in a separate
hutch, and death from general tuberculosis occurred ninety-two days
after inoculation.
The diaphragm and mesentery were studded with tubercles the
size of a pin’s head. The kidneys superficially showed whitish
rounded nodules projecting above the surface. These were found
on section to be continuous, with wedge-shaped deposits in the sub-
stance of the kidney. The lungs presented a very striking appear-
ance, being, in short, a mass of tubercular deposit; and the bronchial
and tracheal glands were similarly affected. In sections of the
kidney and lung the bacilli were present, but they were distributed
irregularly ; in one part of a section it was difficult to detect a
single bacillus, in other parts they were present in large numbers.
The milk from the two cows,
previously to their coming under
observation, had been mixed with
the general supply of a dairy. There
is indeed ample evidence that, both
in this and in other countries, the
milk of tuberculous animals finds its
way into the market. The question
which naturally arises is the possi-
bility of any manifestation of tuber-
culosis in man, arising from the
consumption of unboiled milk con-
taining tubercle bacilli. We must
admit that there is no direct Fic. 173.—TupercuLaR ULcERA-
at! TION OF THE INTESTINE OF A
RaBBit.
evidence of the transmission
tuberculosis by milk from cow to
man; but this may arise from the difficulty in tracing such a
source of infection, owing to the long time which elapses before
symptoms manifest themselves in man. Yet, if milk be a source of
infection, we should naturally expect that primary tuberculosis of the
intestine would be by no means an uncommon manifestation of the
disease; and this in the adult is not in accordance with clinical
experience. Such an argument would tend to contra-indicate
danger to adults; but, on the other hand, the possible danger to
children has been rightly insisted upon by the earliest writers on
this subject. Woodhead has recently stated that, from his experi-
ence in two large hospitals, he has been much struck by the fact
that, in children who had died from other diseases during the course
of tubercular disease of the abdominal glands, there was frequently
396 INFECTIVE DISEASES.
not any trace of tubercular disease in other parts; thus pointing to
the intestine as the channel by which the bacillus made its way into
the body. Woodhead also remarks that in a large number of cases
Fic.’ 174. TUBERCULOSIS OF THE LUNGS.
From a photograph of the lungs of a rabbit which had been injected sub-
cutaneously with about ten drops of milk, including in suspension a small
quantity of the deposit at the bottom of a sample of milk from a cow with
tuberculosis of the udder. Death occurred from general tuberculosis ninety-two
days afterwards. The appearance of the lungs was very striking. They were
almost completely composed of tubercular deposit. The bronchial glands,
as well as the tracheal, of which one is seen in the photograph, were also
enlarged and caseous. There were tubercular deposits in the kidneys and
other organs, and also at the seat of inoculation.
of general tuberculosis, where the possibility of infection by the
pulmonary passages was evidently excluded, the tubercular process
TUBERCULOSIS. 397
appeared to have invaded the body by the intestinal canal. These
facts, taken in connection with the occasional existence of tubercle
bacilli in milk, went far to prove, in his opinion, that milk was a
source of tubercular infection, especially to young children.
From his own experiments and observations the author has
drawn the following conclusions :—
1. Cows with tuberculosis of the udder are to be found in dairies
in this country.
2. The milk of these cows is, as a rule, mixed with the general
supply.
3. The milk in cases of udder tuberculosis contains tubercle
bacilli.
4. Rabbits inoculated with, or fed upon, milk containing tubercle
bacilli contract tuberculosis.
5. Direct evidence of transmission of tuberculosis by milk to man is
wanting, but from the effect of such milk onthe lower animals
it is reasonable to conclude, in the present state of our know-
ledge, that there may be danger in using the milk of cows
with tubercular udders, and therefore strict inspection of
dairies should be enforced; and boiling of milk before
use will, as a rule, be a wise, if not absolutely a necessary
precaution.
Bollinger has shown that the virulence of cow’s milk is reduced
by dilution with water in the proportion of 1 in 40 and even of
1 in 100, and that therefore there would be much less danger
in consuming tubercular milk which had been mixed with the
milk of healthy cows, than there would be in taking it direct
from the infected cow. This is a matter of scientific interest; but
it would be no justification for a dairyman to mix the milk of a
tubercular cow with milk of cows known to be healthy. The milk
of cows suffering from tuberculosis should undoubtedly be rejected.
TUBERCULOSIS AND THE Pusitic Meat Suppty.
The question of the advisability of allowing the flesh of tuber-
cular animals to be sold for food, especially when the disease exists
in a very small degree, is a vexed one. Numerous experiments
have been made upon the infectivity of the flesh of tubercular
animals. Kastner inoculated the juice expressed from the flesh of
tukercular cows, Sixteen guinea-pigs were unaffected after injection
398 INFECTIVE DISEASES.
of 1 to 2 cc. into the peritoneal cavity. Nocard injected ten to
twenty drops of the muscle juice of the hearts of tubercular cattle, in
which the disease was well marked, and none of the guinea-pigs were
infected. With juice of the muscles of the thigh derived from ten
tubercular cows Nocard inoculated forty guinea-pigs, and one only
showed signs of tubercle. Nocard concluded that if there was any
danger in the flesh of tuberculous animals, it was the exception and
not the rule. On the other hand, Chauveau and Arloing produced
tuberculosis in two guinea-pigs out of ten inoculated with muscle
juice from a tubercular steer.
In 1890 a Royal Commission was appointed to investigate this
subject, and the report was issued in 1895. Martin, on behalf of
the Commission, tested the flesh of twenty-one tubercular cows. In
two cases only was evidence obtained of the presence of the bacillus
by inoculation of guinea-pigs. The flesh of eight cows affected with
mild tuberculosis produced tubercle in one instance by inoculation,
but the ingestion experiments were negative. The flesh of five cows
severely affected with tubercle gave the disease in four cases, either
by feeding or inoculation, but only one gave the disease both ways.
Martin thought that some of the results were due to the butcher
infecting the meat in the process of dressing the carcase, either by
his hands or knives. Woodhead made a series of experiments to test
the effects of roasting and boiling on the tubercular virus in meat.
It was found that in boiling and roasting experiments, as ordinarily
carried out in the kitchen, the temperature, however high it may be
on the surface, seldom reaches 60° C. in the centre, except in the
case of joints less than about six pounds in weight. Boiling and
roasting were found insufficient to destroy tubercular virus enveloped
in rolls of meat.
The following were among the conclusions of the Commissioners :—
We have obtained ample evidence that food derived from tuberculous
animals can produce tuberculosis in healthy animals. The proportion of
animals contracting tuberculosis after experimental use of such food is
different in one and another class of animals; both carnivora and
herbivora are susceptible, and the proportion is high in pigs. In the
absence of direct experiments on human subjects, we infer that man also
can acquire tuberculosis, by feeding upon materials derived from tuber-
culous food-animals.
The actual amount of tuberculous disease among certain classes of
food-animals is so large as to afford to man frequent occasions for
contracting tuberculous disease through his food. As to the proportion
of tuberculosis acquired by man, through his food or through other means,
we can form no definite opinion, but we think it probable that an
TUBERCULOSIS. 399
appreciable part of the tuberculosis that affects man is obtained through
his food.
The circumstances and conditions with regard to the tuberculosis in
the food-animal which lead to the production of tuberculosis in man are,
ultimately, the presence of active tuberculous matter in the food taken
from the animal, and consumed by the man in a raw or insufficiently
cooked state.
Tuberculous disease is observed most frequently in cattle and in
swine. It is found far more frequently in cattle (full grown) than in
calves; and with much greater frequency in cows kept in town cow-
houses than in cattle bred for the express purpose of slaughter. Tuber-
culous matter is but seldom found in the meat substance of the carcase ; it
is principally found in the organs, membranes, and glands. There is
reason to believe that tuberculous matter, when present in meat sold to
the public, is more commonly due to the contamination of the surface
of the meat with material derived from other diseased parts, than to
disease of the meat itself. The same matter is found in the milk of cows
when the udder has become invaded by tuberculous disease, and seldom
or never when the udder is not diseased. Tuberculous matter in milk is
exceptionally active in its operation upon animals fed either with the
milk or with the dairy produce derived from it. No doubt the largest
part of the tuberculosis which man obtains through his food is by means
of milk containing tuberculous matter.
Provided every part that is the seat of tuberculous matter can be
avoided and destroyed, and provided care be taken to save from contami-
nation by such matter the actual meat substance of a tuberculous
animal, a great deal of meat from animals affected by tuberculosis may
be eaten without risk to the consumer.
Ordinary processes of cooking applied to meat which has got con-
taminated on its surface are probably sufficient to destroy the harmful
quality. They would not avail to render wholesome any piece of meat
that contained tuberculous matter in its deeper_parts. In regard to milk
we are aware of the preference by English people for drinking cow’s milk
raw—a practice attended by danger, on account of possible contamination
by pathogenic organisms. The boiling of milk, even for a moment, would
probably be sufficient to remove the very dangerous quality of tuber-
culous milk.
TUBERCULOSIS IN EQUINES.
Tuberculosis is not very common in the horse, but when it does
occur, it is frequently mistaken for glanders. There may be miliary
tuberculosis in the lungs, or nodules disseminated throughout the
lungs, liver, spleen, and bones. In a number of cases investigated
by Nocard, the disease commenced in the abdominal organs, and
the affection of the lungs appeared to be secondary. The author
has examined several cases of equine tuberculosis. In some cases
400 INFECTIVE DISEASES.
the lungs were affected with the disease in a miliary form. The
bacilli could not be distinguished from bacilli in sections of the bovine
disease. Giant cells were extraordinarily numerous, and in many
cases were densely packed with bacilli, so that they could be recog-
nised en masse under a low power. The bacilli were also distributed
in the tissue generally, but were much more numerous in the giant
cells.
TUBERCULOSIS IN Dogs.
Peters described a case of tuberculosis in a pet dog, from eating
sputum from a tubercular patient. This is said to be a not
uncommon cause of canine tuberculosis.
TUBERCULOSIS IN Cats.
Nocard reported a case of tuberculosis in a cat from eating tuber-
cular sputum. The abdominal organs were diseased. Bollinger
has described two cases of miliary tuberculosis. M‘Fadyean also has
described a case. The bacilli are very plentiful in the lung. A
minute examination of the individual micro-organisms by the author
did not reveal any distinctive character.
TUBERCULOSIS IN SWINE.
The author examined the tubercular liver of a pig. The pig
was about six months old, and after suffermg from cough and
emaciation, died.
The liver had caseous nodules scattered throughout its substance,
some the size of a pea, and others larger. Tubercle bacilli without
distinctive characters were found on examination of sections; but
it was in some parts of a preparation difficult to detect any bacilli,
and in other parts there were not more than five or six in the
field of the microscope. Tuberculosis in swine is said to be very
rare in America.
TUBERCULOSIS IN Brrps.
Hens, guinea-fowls, turkeys, pheasants, and partridges, are sub-
ject to tuberculosis, and ostriches and other birds kept in confinement
may contract the disease.
Tuberculosis in fowls appears to be introduced principally with
the food, the disease occurring commonly in the intestines and
liver.
DESCRIPTION OF PLATE XIII.
Tuberculosis in Swine.
Section of liver of a pig with scattered tubercular nodules. Microscopica:
sections of the liver showed tubercle bacilli in very small numbers.
th a FD
“‘OId AO AAAIT UVITINOY
Sa.
Tah.
Be pec Loe re aa paper ie
TUBERCULOSIS. 401
The author has examined several cases of so-called spontaneous
tuberculosis in fowls. Sections of the liver were in one case remark-
able on account of the extraordinary invasion of the caseous deposits
with bacilli. Cover-glass preparations had been made from the
liver in the following way for diagnostic purposes: A tubercle was
readily picked out on the point of a scalpel and crushed between two
slides, and the cover-glass preparations stained with the Ziehl-
Neelsen solution. The bacilli are for the most part very small. A
few attain a considerable length, but the majority are in the form
of small, straight rods, with many sizes intervening between. these
rods and isolated granules.
In July 1888 the author received from Mr. Bland Sutton the
liver and lungs of a Rhea, which had died in the Zoological Gardens.
The lung was infiltrated with caseous deposits, and there were
scattered caseous nodules in the liver varying in size from a pea
to a marble. The naked-eye appearance of a section of the liver
through these nodules, at once recalled to mind the naked-eye
appearance of the deposits in the pig’s liver already described. But
* whereas in microscopical preparations of the pig’s liver, bacilli were
very scantily present, the sections of the lung and liver of the Rhea
contained bacilli in such extraordinary numbers that, under a power
of fifty diameters, the collections of bacilli could be recognised as red
granular masses. These red masses under a high power were re-
solved into dense colonies of bacilli. In their number and their
distribution in the tissues, in their varying size, and in the extra-
ordinary length of the longest forms, they presented very interesting
points for observation. From the naked-eye appearance of the
disease and the general microscopical characters, as well as the
presence of bacilli agreeing in their staining reactions with the
classical tubercle bacilli, the author had no hesitation in pronouncing
the disease to be avian tuberculosis.
Klein, who had examined a similar case, alluded to it in a
description of leprosy; but this disease is unknown in the lower
animals, and all attempts to infect them from man have been
almost, if not entirely, negative.
The bacilli in the Rhea are principally collected in the caseous
parts, but they are also found in the tissue generally, and often
collected in large cells. In size they vary toa marked extent. In
the cells they often form compact masses of short bacilli, but in
other parts, both in collections and singly, they attain a greater
length than is observed in any other form of tuberculosis. Some of
the bacilli present a very interesting appearance. They are provided
26
402 INFECTIVE DISEASES.
terminally with a sharply defined ovoid body. There are also
collections of short bacilli, many with these spore-like appearances.
The author has also seen free ovoid forms, sometimes singly, some-
times in groups. From their connection with the bacilli and their
sharply defined outline they are very suggestive of spores.
Johne examined the livers of a number of fowls accidentally
infected by phthisical sputum. Nocard reported an outbreak in
a poultry-yard where the man in charge had consumption. He
also found the disease amongst fowls fed with the infected organs
of tubercular cattle. Subcutaneous inoculation, and feeding of fowls
with sputum or bovine virus, will produce the disease.
Experimental inoculation of. tubercular virus from different
sources affords an illustration of the different pathogenic effects
obtained by varieties of the same species of bacillus. The bacillus
of fowl-tuberculosis is a distinct variety. A very small proportion
of guinea-pigs, inoculated in the peritoneal cavity with fowl-tubercle,
succumb to the disease, though so susceptible to the effects of human
or bovine virus. Maffucci maintains that guinea-pigs have an
immunity, and that rabbits rarely develop a generalised tuberculosis.
Cultures are not identical in appearance with those obtained from
man, and on microscopical examination show many long, thick, and
branched forms, which are only rarely found in cultures from a
human source.
Stamping-out System.—In 1888 a Departmental Committee
was appointed to inquire into pleuro-pneumonia and tuberculosis,
and they considered that legislation ought to be directed not only
to the protection of cattle from tuberculosis, but also to prevent
the possibility of the disease being communicated to man.
The following extracts are from the recommendations of the
Committee, which were made on the lines of :—
A. PREVENTION.
B. Exrirparion.
A.—Preventive Measures.
These should include provision for :—
Improved hygiene of cattle sheds, etc. (especially in the direction of
providing proper ventilation, pure water supply, and adequate disinfection
of stalls, etc., wherein tubercular animals have been kept). This has
been partly met in the Dairy and Milk Shops Order, but its administra-
tion by the local health authorities is at present imperfect ; and we would
suggest that it should be much more stringently enforced, and that
veterinary inspectors should be given more extended powers of entry into
all places where animals are kept.
TUBERCULOSIS, 403.
Improvement in the hygienic surroundings of animals should include
isolation of all suspected cases, precautions against the flesh or milk of
diseased animals being given as food to others—e.g., to pigs, fowls, etc.—
and care that fodder, litter, and water should not be taken from one
animal or stall and given to ‘annthen.
Our attention has been drawn to the frequency with which animals,.
obviously diseased, sometimes even in the last stage of the malady, are
sold in open market.
Although in England and Ireland, under the provisions of the
Nuisances Removal Act as embodied in the Public Health Act, 1885,
the medical officer of health or inspector of nuisances may seize such
animals, yet such seizure is rarely performed.
We find the veterinary inspector has no power to prevent such sales,
or to seize the beasts for slaughter, since tuberculosis is not included in
the Contagious Diseases (Animals) Act of 1878.
We further find that there is actually a regular trade in such stock
infected with tuberculosis, and that they go by the name of “ wasters”
and “ mincers,” being frequently slaughtered in the neighbourhood of the
larger towns, to which such portions of the meat as are likely to escape
the observation of the inspector of nuisances are sent, for the purposes
of sale among the poorer inhabitants, and especially for the making of
sausages.
We are, therefore, very strongly of opinion that power should be given
to the veterinary inspector to seize all such animals in fairs, markets, or
in transit.
Notwithstanding the uniform prevalence of the disease in Europe and
elsewhere, there seems to be no reason to apprehend that, with our
present regulations for the slaughter of animals at the port of debarka-
tion, and for quarantine of those imported for breeding, there is any
special danger of increasing the infection in England by introduction
from abroad. The danger, however, exists in regard to the stock brought
from countries, which are exempt from slaughter on landing, and sub-
jected to the ordinary veterinary inspection during the present period of
detention of twelve hours.
It is, therefore, evident that the present rules for the prevention of
‘the introduction of disease into the United Kingdom from abroad, are
incomplete.
Since all authorities are agreed that the disease is very marked by
heredity, we think it highly desirable that breeders should in their own,,
as well as in the public interest, discontinue breeding from tuberculous.
stock.
B.— Extirpation.
In order to insure the gradual extirpation of tuberculosis, we are of
opinion that it should be included in the Contagious Diseases (Animals)
Acts for the purposes of certain sections of those Acts, so as to provide :—
(a) For the slaughter of diseased animals, when found diseased on
the owner’s premises.
404 INFECTIVE DISEASES.
(b) For the payment of compensation for the slaughter of such
animals,
(c) For the seizure and slaughter of diseased animals exposed in fairs,
markets, etc., and during transit.
(d) For the seizure and slaughter of diseased foreign animals at the
place of landing in this country.
Notification of this disease should not be compulsory, because it may
exist without developing any sufficient outward evidence to enable the
owner to detect it, and its growth is so slow, that non-notification of its
existence, even in a large number of cases, would do little to nullify the
stamping-out effect of the Act of 1878.
The powers and responsibilities of inspectors in ordering the slaughter
of diseased animals should be the same for tuberculosis as for pleuro-
pneumonia, according to section 51 (5) of the Act of 1878.
Further, tubercle, though hereditary, is nevertheless much less conta-
gious than the other diseases included under the Act of 1878, and it is
clear, therefore, that the immediate slaughter of diseased animals would
go far to stamp it out, though, doubtless owing to heredity, this stamping-
out process would be gradual in its effect.
A supplementary report was made by Professor Horsley, in
which he expressed the opinion that there ought to be legislation
to prevent breeding from diseased animals, and compulsory notifi-
eation :—
1. Breeding.
Tuberculosis is notorious, even among the laity, as a disease which is
transmitted from parent to offspring. This is a fact with which cattle
breeders are specially familiar, and which finds strong expression in the
evidence attached to this report. Further, this generally received truth
has been completely confirmed by the results of scientific investigation,
as is also duly set forth in the report. Considering, therefore, the
extreme importance of this point, I think that the act of wittingly breed-
ing from animals so affected should be made an indictable offence. The
only objection that can be raised to such legislation, which if effected
would prevent the dissemination of the disease among cattle in this
country, is that, owing to the present state of want of knowledge among
cattle owners, and even veterinary surgeons, of the early symptoms, and
physical signs on examination, of this disease, prosecutions would occa-
sionally occur in cases in which no fault could properly be attributed to
the owner, and that, therefore, such prosecutions would be needlessly
vexatious.
Considering, however, the extreme rarity with which such cases would
occur, and that, as in the matter of non-notification, each case would be
tried before district magistrates on its own merits, this objection is
deprived of the force it might have possessed.
_ TUBERCULOSIS. 405
2. Notification of the Existence of the Disease.
This point requires no explanation, since it is clear that, unless the
veterinary inspectors or authorities receive information of occurrence of
diseases, it is impossible to ensure the thorough carrying out of the
provisions of the Contagious Diseases (Animals) Act.
That deliberate non-notification should be punished cannot be doubted
by any one. Objection, however, to legislation in this direction has been
put forward, on the same grounds as those upon which the prevention of
breeding from diseased animals was contested. As, however, I consider
that these objections have been already shown to have no weight, I
recommend that both the forbiddance of breeding from diseased animals,
and the notification of the disease, should be included in any legislation
for tuberculosis.
The difficulty referred to by the Committee, is presented by
cases of the disease which cannot be detected by the ordinary
methods of examination, and might possibly be overcome by the
use of tuberculin as a diagnostic agent.
CHAPTER XXIX.
LEPROSY.—SYPHILIS.—RHINOSCLEROMA.— TRACHOMA,
LEpRosyY.
Leprosy occurs in three forms—tubercular, anesthetic, and mixed
tubercular. It may be classed with the granulomata, as the most
common form of the disease is characterised by deposits in the skin,
mucous membrane, and internal organs. These deposits are composed
of small cells, and large cells resembling giant cells. The cells
become deposited in the surrounding tissues, and so the tubercle
enlarges, involving the epidermis and developing into an ulcerating
sore; or, after a certain stage of development, beginning to decline,
and finally leaving a puffy discoloration. In the anesthetic form
the cells invade the connective tissue of nerves. In the mixed
form the varieties occur together, but the tubercular character
predominates.
Tubercular leprosy commences with the development of an
erythematous patch, which becomes infiltrated, and finally tubercu-
lated, the tubercles varying in size from a millet seed to a marble,
or even larger. The eruption on the head and face produces a
characteristic leonine expression. The progress of the disease is very
slow. After death the following changes may be found in the
internal organs: Cirrhosis of the liver and spleen, enlargement of
the lymphatic glands, and a condition of the lungs corresponding
to cheesy bronchial pneumonia.
In the anesthetic form patches develop on the skin, which
become anesthetic; ulceration follows, and the fingers and toes, or
the entire hand and foot, may slough off.
The disease is undoubtedly communicable, but the infectivity is
of a very low type.
The infectiousness is illustrated by the well-known case of Father
Damien. Arning inoculated a man named Keanu, a condemnéd
criminal, and leprosy developed three years afterwards, but this case
406
LEPROSY. 407
is not regarded as conclusive, as the man had a family history of
leprosy. The disease has never been known to spread from patients
in this country, who have contracted the disease abroad.
The bacilli of leprosy were first observed by Hansen in 1874, and
subsequently fully described by him, and his observations confirmed
by Neisser, in 1879.
Bacillus Lepree.—Rods 5 to 6 » in length and 1 » in breadth.
The bacilli are straight or curved, resembling very closely the
tubercle bacilli. They are present in the leprous tubercles of the
skin and mucous membrane, in the lymphatic glands, and in the
liver, testicles, and kidney; and in the nerves in the anzsthetic
variety. They are found between the cells, and in colonies in the
cells. They stain readily with the aniline dyes, especially by the
Ziehl-Neelsen and Gram’s methods. The bacilli are found in extra-
ordinary numbers in the skin, and they are rather straighter than
tubercle bacilli, and stain more readily.
Numerous unsuccessful attempts to cultivate the bacillus have
been made by many bacteriologists. The author has made repeated
inoculations upon glycerine-agar, upon which the tubercle bacillus
grew abundantly, but always with disappointing results. On the
other hand, Bordoni-Uffreduzzi showed the author a cultivation
which he had obtained from the bone marrow of a leper. The
cultivation was made on blood serum and glycerine, and cover-
glass preparations resisted decolorisation with acid. There were
slight morphological differences when compared with the appear-
ance of bacillus lepre in the tissues, and the results were hardly
conclusive.
The English Leprosy Commission also reported successful cultiva-
tion of the leprosy bacillus. The author had the opportunity of
examining one of the first cultures received in this country, and
found that the bacilli stained deeply in ordinary cover-glass pre-
parations, they did not resist decolorisation by the Ziehl-Neelsen
method, and they corresponded in culture with one of the varieties
of Bacillus subtilis, commonly found on the skin.
Inoculation of animals has given equally unsatisfactory results.
Numerous experiments have been made by Beaven Rake on small
animals and birds, with invariably negative results. The blood of
leprous patients, tubercles from the living subject, fragments of the
skin and of the internal organs after death, have been inoculated by
different observers without result. Melcher and Ortmann alone claim
to have produced really definite results. These observers excised
leprous tubercles from the living subject, and inoculated fragments
408 INFECTIVE DISEASES.
in the anterior chamber of the eye of rabbits. The animals died
after some months with extensive deposits in the cecum, lymphatic
glands, spleen, and lungs.
These tubercles varied in size from a pin’s head to a millet seed,
and contained bacilli, resembling leprosy bacilli in their staining
reactions. The question naturally arises whether the lesions were
really indicative of leprosy or tuberculosis. Until the experi-
ments are independently confirmed, and the result of inoculation
differentiated from tuberculosis, it would be rash to accept these
experiments as conclusive.
It has been suggested that tuberculosis and leprosy are identical.
There is a similarity in the bacilli and in the lesions of leprosy and
tuberculosis, the injection of tuberculin produces a reaction in leprosy
nodules, and many lepers die from tubercular disease of the lung.
But while tuberculosis is very readily transmitted to guinea-pigs
and rabbits by inoculation of fragments of tubercular tissue, leprosy
is inoculable, if at all, in most exceptional instances. The bacilli
of tubercle are cultivated with the greatest facility, the bacilli of
leprosy, if at all, only with exceptional difficulty; tubercle bacilli
are found in giant cells, leprosy bacilli in the so-called leprosy
cells. Leprosy bacilli are straighter than human tubercle bacilli,
and differ slightly in their behaviour to staining reagents. On
the other hand, the morphological differences are not greater than
those existing between different forms of tubercle bacilli obtained
from tuberculosis in animals and birds. It would be premature
to regard leprosy as a variety of tubercle until cultivations of
the bacillus have been obtained, and carefully compared with
those of the tubercle bacillus. Differences in morphological details
and results of inoculation would then carry less weight as a means
of differentiation.
The tubercular pneumonia of lepers would be regarded, if the
bacilli are identical, as a development of leprosy in the lungs, and
not, as at present, a result of double infection with tuberculosis.
Mernops or Examinine tHe Bacitius oF LEpRosy.
Cover-glass preparations may be made in the ordinary way, or by a
special method, which consists in clamping a nodule with a pile clamp.
until a state of anemia of the tissue is produced. On pricking with a
needle or sharp knife a drop of clear liquid exudes, from which cover-
glass preparations may be made, and stained by Neelsen’s method.
For sections the author prefers Neelsen’s method and methylene-blue.
They can also be stained by Gram’s method, which, as a rule, brings out
very clearly the beaded appearance of the bacilli.
DESCRIPTION OF PLATE XIV.
Bacillus Lepr.
Fic. 1.—From a section of the skin of a leper. The section is, almost in
its entirety, stained red, and, with moderate amplification, has a finely
granular appearance. Stained by the Ziehl-Neelsen method (carbolised
fuchsine and methylene-blue). x 200.
Fie. 2.—Part of the same preparation with high amplification, showing that
the appearances described above are due entirely to an invasion of the
tissue by the bacilli of leprosy. x 1500.
Plate XIV.
BACILLUS LEPRZ
LEPROSY. 409:
Method of Babes.—Preparations are stained in rosaniline hydrochlorate
in aniline water, decolorised in 33 per cent. hydrochloric acid, and after-
stained with methylene-blue.
Stamping-out System.—The history of leprosy in the British
Islands during the Middle Ages, and the conditions under which it
both increased and declined, have been discussed by several writers.
A large number of institutions of a charitable and ecclesiastical
character were established in endemic areas and were occupied by
the lepers either voluntarily, or compulsorily by means of the Act
De leproso amovendo. These institutions were to a very small extent
a means of segregation. According to Dr. Newman the disease,
which had reached its zenith about the twelfth or thirteenth century,
began to decline from that time owing to “a general and extensive
social improvement in the life of the people, to a complete change
in the poor and insufficient diet (which it is evident consisted far
too largely of bad meat, salt, putrid and dried fish, and an almost
entire lack of vegetables) and to agricultural advancement, improved
sanitation and land drainage.” Of all the unfavourable conditions
it would appear that food in some way was especially associated with
the cause of the disease, either by introducing the bacillus or by
rendering the tissues a suitable soil for its reception and development.
In other countries segregation has been attempted voluntarily
or compulsorily, but it has never been completely carried out. There
can be very little doubt that the presence of a leper in a healthy
community is no greater source of danger than the presence of
an individual suffering from tuberculosis, but, for other reasons,
voluntary isolation should be carried out as completely as local
circumstances will permit.
The Leprosy Commission in India recommended—
(a) That the sale of articles of food and drink by lepers should
be prohibited, and that lepers should be prevented from
following certain specified occupations.
(6) That the concentration of lepers in towns should be discouraged.
(c) That Leper Asylums should be established in which lepers
might live voluntarily.
(d) That Leper Farms scattered over the country should be
encouraged.
(e) That the few children who are born of lepers should be
removed to Orphanages.
They concluded that by means of improved sanitation and good
dietetic conditions a diminution of leprosy will result.
‘
410 INFECTIVE DISEASES.
SYPHILIS.
Syphilis is a disease peculiar to man, and communicable ouly by
inoculation. The local infection is followed by a period of latency,
and by a period during which generalised eruptions appear. One
attack confers immunity from future attacks. The virus in its
most virulent form is found in the primary seat of inoculation, and
in the indurated glands which
follow. It is also supposed to be '
present in the blood and secretions.
Lustgarten, Eve and Lingard have
found bacteria which they believed
to be specific.
Bacillus in Syphilis (Lustgar-
ten).—Rods resembling the bacilli
of leprosy and tuberculosis, 3 to 4 p
Fic. 175. — CovER-GLASS PREPARA-
TION OF Pus FROM A CHANORE, X long, “8 p thick. ; Two or more
1050 (LUSTGARTEN). colourless, ovoid points in the course
of the rod are visible with a high
power; it is thought that possibly they are spores. The bacilli
are always found in the interior of nucleated cells, which are
more than double the size of
leucocytes. They have been ob-
served in the discharge of the
primary lesion, and in tertiary
gummata.
Alvarez and Tavel state that
an identical bacillus is found in
normal secretions (smegma). Eve
and Lingard have described a bacil-
lus associated with specific lesions,
which differs from the above in its
morphology and behaviour towards Fic. 176.—WanprRING CELL con-
TAINING BaciLi1 (LUSTGARTEN).
staining reagents.
Meruops oF Srarning THE Bacituus oF SYPHILIS.
Method of Lustgarten :—
Sections are placed for twelve to twenty-four hours in the following
solution, at the ordinary temperature of the room, and finally the solution
is warmed for two hours at 60° C. :—
Concentrated alcoholic solution of gentian-violet . 11
Aniline water . ; ‘ F : : 100
SYPHILIS. All
The sections are then placed for a few minutes in absolute alcohol,
and from this transferred to a 1:5 per cent. solution of permanganate of
potash. After ten minutes they are immersed for a moment in a pure
concentrated solution of sulphurous acid. If the section is not completely
decolorised, immersion in the alcohol and in the acid bath must be
repeated three or four times. The sections are finally dehydrated with
absolute alcohol, cleared with clove-oil, and mounted in Canada balsam.
By this method the bacillus is distinguished from many bacteria, but
not from the bacilli of tubercle and leprosy which are stained by this
process.
Method of De Giacomi :—
Cover-glass preparations are stained with hot solution of fuchsine
containing a few drops of perchloride of iron. They are then decolorised
in strong perchloride of iron, and after-stained with vesuvin or Bismarck-
brown.
Method of Doutrelepont and Schiitz :—
Sections are stained in a weak aqueous solution of gentian-violet and
after-stained with safranin.
The nature of the contagium in syphilis is unknown.
Protective Inoculation.—Incculation of the virus, or syphilisa-
tion, as a protective measure, was at one time practised and strongly
advocated; but it is rightly regarded in this country as dangerous
and unjustifiable. From the experiments of Ricord it would appear
that the local results in the vesicular stage resemble the results of
the inoculation of virulent vaccinogenic grease or horse-pox. The
inoculation goes through the stages of papule, vesicle, ulcer, scab,
and scar. The accidental inoculation which occurs in cases of
vaccino-syphilis may so closely resemble the results of inoculation
with very virulent cow-pox, that it is sometimes difficult to decide
as to the exact nature of these cases.
RAINOSCLEROMA.
Rhinoscleroma is a rare disease, resembling lupus, and pro-
ducing in the nostrils and neighbouring parts nodular swellings,
composed of granulation-tissue. The disease is met with in
America, Egypt, Austria, and Italy. There are no giant cells, but
peculiar large cells, which were first described by Mikulicz. Frisch
discovered bacteria in sections, and Cornil and Alvarez pointed out
the existence of a capsule. In morphology and cultivation they
resemble, according to Dittrich, Friedlander’s pneumococcus. They
are probably identical with this micro-organism, and Paltauf and
Eiselsberg, and others, found that they produced septicemia in rabbits
and guinea-pigs.
Bacterium of Rhinoscleroma (Bacillus of Rhinoscleroma,
412 INFECTIVE DISEASES.
Cornil and Alvarez).—Cocci and short rods, 15 to 3 yw in length,
‘5 to ‘8 p thick. Deeply coloured points or granules may occur
in the course of the rods when stained, but it is very doubtful
whether these can be considered as spores. The bacteria are en-
capsuled, the capsule being round when enclosing a coccus, and
ovoid when enclosing a rod. The capsule is composed of a tough
resisting substance; two or more capsules may unite by fusion,
enclosing two, three, or a greater number of rods. The bacilli were
observed in sections of the tumours, which developed on the lips
and in the nasal and pharyngo-laryngeal regions.
Mernop or Stainine THE BactLius oF Ruaino-ScLERoMmA.
Methad of Cornil and Alvarez :—
Sections are immersed in a solution of methyl-violet (B) for twenty-
four to forty-eight hours, with or without the addition of aniline-water ;
and are then decolorised after treatment with the solution of iodine in
iodide of potassium. If the sections are left to decolorise in alcohol for
forty-eight hours, the capsule is rendered visible.
TRACHOMA.
Trachoma is a disease of the conjunctiva, common in Egypt.
The new growth is composed of round cells, and may be regarded,
according to Kartulis, as the chronic stage of either gonorrheeal or
Egyptian ophthalmia. Koch failed to find any micro-organisms
in the swollen lymph follicles. Sattler asserted that he had culti-
vated a micrococcus which produced the disease when inoculated
on the conjunctiva, Other observers have found the common
pyogenic micrococci in the secretions, especially Staphylococcus
pyogenes aureus and albus.
CHAPTER XXX.
ACTINOMYCOSIS.—MADURA DISEASE.
ACTINOMYCOSIS.
Actinomycosis belongs to the class of infective granulomata. It is
a chronic inflammatory affection characterised by the presence of a
special microphyte, which by irritation produces a neoplasm, composed
of round cells, epithelioid cells, giant cells, and fibrous tissue. These
neoplasms form nodular tumours of various sizes. In some cases
there is a tendency to develop very large tumours, and in others to
break down early and suppurate. In cattle, cretification takes
place in the fungus tufts. Actinomycosis closely resembles tuber-
culosis in its histological characters. The disease attacks man,
horses, cattle, and pigs.
Many interesting observations have been made upon the origin
of this disease in man. Two cases have been recorded in support of
the theory of direct infection from the cow. Stelzner described a
case of actinomycosis in a man who had had the care of animals,
some of which had suppurating glands. Hacker had a case of
actinomycosis of the tongue in a man who had charge of cows, one
of which had a tumour of the jaw which he had opened. On the
other hand, Moosbrugger found that out of 75 cases, 54 were in men,
and 21 in women, including 2 children. In 11 of these men the
occupation was not stated. In 33 their occupation did not bring
them into contact with diseased animals; they were, for example,
millers, glaziers, tailors, shop people, and students. Only 10 cases
occurred among farmers, peasants, and farm-labourers, and in only
one case out of the 10, had the patient been brought into contact
with diseased animals.
Out of the 21 women, there were only 4 peasants, and none
of them had been associated with diseased cattle.
Infection by the flesh of diseased animals has also been dis-
cussed. But there is no evidence of prevalence of the disease
413
414 INFECTIVE DISEASES.
among slaughterers and butchers, who would be particularly liable
to it, if flesh were a source of infection. The chances of infection
by ingestion are minimised by the flesh being almost always cooked.
Actinomycosis occurs also in pigs, and pork is very often eaten in an
uncooked state; but Israél has pointed out that this may probably
be excluded, as many of the cases occurred among strict Jews.
The evidence points to the disease originating in man and lower
animals from the same source, and there is a very strong suspicion
attached to cereals. This view is supported by important obser-
vations, with reference to the part played by cereals in inducing
the disease in cattle, and it gains additional support from a case
described by Soltmann, where the disease resulted from an awn of
wall barley. A boy, aged eleven, accidentally swallowed an awn
of Hordeum murinum. He became very ill, and suffered great pain
behind the sternum, extending to the back. An abscess formed,
covering an area extending over six intercostal spaces, and when
opened, the awn of this grass was found in the evacuated pus.
The pain, however, continued, and fresh deposits occurred, and when
the boy was taken to the hospital, the ray-fungus was detected.
Possibly the spores of the fungus can be conveyed both by air and
water.
This disease in cattle has long been known in this country, but.
its various manifestations were either mistaken for other diseases,
or simply received popular names. Indeed, the various forms are
still familiar to many as wens, clyers or crewels, scrofulous, tuber-
cular or strumous abscesses, polypus, lymphoma, cancer of the
tongue, scirrhous tongue, indurated tongue, ulcerated tongue, cancer
of bone, bone tubercle, osteo-sarcoma, fibroplastic degeneration of
bone, spina ventosa, and carcinoma.
Bovine antinomycosis is especially prevalent in river valleys,
marshes, and on land reclaimed from the sea. The disease occurs.
at all times of the year, but general experience leads to the belief
that it occurs more commonly in the winter.
It is more frequently met with in young animals, and usually
occurs between one and three years, but it may be found at almost
any age, and probably affects equally both sexes.
There is little if any evidence to show that the disease is heredi-
tary. In numerous cases, the family history has been most carefully
inquired into by the author; and in the case of some imported
pedigree animals, the disease was quite unknown on the farm where
they had been bred.
The tongue is so commonly the seat of the disease, that suspicion.
ACTINOMYCOSIS. 415
at once falls on food as the means by which the parasite is conveyed.
Skin wounds produced by rubbing against the mangers, posts, or
wire fencing, may also become infected.
The evidence is very strong in favour of believing that the micro-
organism gains access to the system through wounds or lacerations
of the mucous membrane and skin, or through carious teeth. It
has been pointed out that the common occurrence of the disease
at the time of the second dentition may be owing to the wounds
produced in the alveolar mucous membrane by the shedding of
the teeth. Experience also points to straw being sometimes a
factor in the production of the disease, and it is possible that thistles
and frozen roots also, by wounding the mucous membrane, may
afford a way for the entrance of the micro-organism. The disease
in the jaws, both in man and in cattle, is very commonly associated
with carious teeth.
The cowsheds, pastures, and drinking tanks may become infected
with the discharges from diseased animals. The discharge con-
taminates the fodder in the sheds, and falls on thistles and siliceous
grasses in the pasture, which may first wound, and then introduce
the micro-organism. The discharge is also coughed out of the
mouth, and expelled from the nose, in cases in which a tumour in
the pharynx, or the nasal chambers, has undergone suppuration.
Jensen believed that the disease was produced by different kinds
of grain, especially when cultivated on ground reclaimed from the
sea. He mentions an instance of a farm, where nearly the whole
of the young stock, about thirty in number, had actinomycosis after
feeding on mixed forage, grown on a certain field. Two years after-
wards the same disease occurred in the same stalls in four animals,
after being fed on barley-straw from the same field. According to
Jensen, the fungus grows on grain, husks, and straw of different
cereals, but most abundantly on barley, which is also the most
likely to wound the mucous membrane. Johne’s observations tend
to corroborate this view, for in twenty-two out of twenty-four cases
in which he found barley sticking in the tonsils of pigs, he found
the beard thickly beset with a fungus very similar to, if not identical
with, the ray-fungus: These observations are of great interest in
connection with Soltmann’s case.
Experience points to the belief that the disease is not readily
communicable from animal to animal, and it is possible that when
it affects a large number of cattle in a herd, the same causes have
been acting to produce the disease in a number, which in another
instance may only produce it in one. At the same time, isolated
416 INFECTIVE DISEASES.
cases are possibly not quite so common as they are reported to be.
It is well known that, as a rule, the services of a veterinary surgeon
are not called for except in hopeless or very severe cases. The
cowmen themselves, in many districts, treat the cows successfully,
and then send them into the market, and thus the existence of
previous cases may not have come to the knowledge of the veterinary
surgeon.
Historical.—In 1845 Professor von Langenbeck, of Kiel, made
notes of a ease of vertebral caries in a man, and prepared drawings
of peculiar bodies in the pus from an abscess. The drawings were
published together with a reference to the case by Israél in 1878.
There can be little doubt that these structures were the fungi of
actinomycosis. But the first to publish observations was Lebert
in 1848,
Lebert received from M. Louis some pus, of a thick, almost
gelatinous consistency, which had been obtained from an abscess of
the thoracic wall in a man aged fifty. The patient had been attacked
four months previously by a pulmonary affection, which was
suspected by M. Louis to be cancerous in nature. The pus contained
a very considerable number of little spherical bodies of a slightly
greenish-yellow colour, about the size of a pin’s head. They
could be readily crushed between two strips of glass, and on
examination with a power of fifty diameters two elements could be
distinguished: a soft connective substance, and many hard, narrow,
wedge-shaped corpuscles, arranged in a radiating manner. Under
a high power these bodies were observed to be 2, to 4, of an inch
in length, y§> in width at the base, and ;1, in width at the
apex. Some of these corpuscles were regular, while others showed
one or two constrictions, with intermediate flask-shaped swellings.
Lebert tested these structures with reagents, with the following
results. The bodies were found to remain unaltered by concentrated
mineral acids. Acetic acid freed them . from foreign elements
adhering to their surface. Solution of caustic potash did not affect
them if used cold, but a boiling solution reduced the cuneiform
structures to a fine greyish powder without dissolving them. Ether,
alcohol, and chloroform had no effect upon them when used either
hot or cold. Solution of potash, in which these bodies had been
heated, mixed with a solution of sulphate of copper and brought to
boiling point, did not offer any uniform red colour, which would
have been the case if they had contained albumin. Thus, the chief
chemical characters of albuminous and fatty substances were
wanting, and they resembled chitine in their behaviour to reagents.
ACTINOMYCOSIS. 417
Lebert bore in mind the possible existence of some helminthic débris,
of which these bodies might be hooklets, but he sought in vain for
echinococci and cysticerci.
Actinomycotic pus was later described and figured by Robin.
In the illustration accompanying the description, the fungi are most
Fic. 177.—Srction or Liver rrom A CAsE OF ACTINOMYCOSIS 1N Man.
accurately depicted. Robin states that he had found, in two or
three cases in the pus of deep-seated chronic abscesses, yellowish grains
attaining a diameter of one-tenth of a mm., surrounded by a sort of
halo or thin, viscous, finely granular stratum, containing leucocytes.
These rains were composed of elements 2 to 6 mm. in length, swollen
5 te)
27
2
A18 INFECTIVE DISEASES.
at one end and tapering off at the other, arranged in a regular series,
radiating from a common centre which consisted of granular matter,
‘They were highly refractive, possessed a brilliant centre and sharply
defined outline ; they were dissolved, or at least rendered indistinct,
by acetic acid, and proved insoluble in ammonia and ether.
The disease in man was next described by Israél in the paper
mentioned above. Ponfick was the first to clearly recognise the
identity of the disease in man with the disease in cattle, and he
described a number of cases in man. Israél subsequently published
a work on the subject. The various cases which had been observed
up to that date were described, and the disease classified according
to the seat of invasion.
From this time onwards numbers of cases in man have been
described, and various important researches published, of which
those of Bostrém and Moosbrugger may be especially mentioned.
In England, Acland recognised a case on examining the liver
after death (Fig. 177). H. Taylor was the first,.in this country,
to detect the fungus during the life of a patient. Shattock found
Specimens of the disease in museums. Skerrit, Powell and Godlee,
Eve, Delepine, Ransome, Poore, Malcolm Morris and others have
published cases.
In Italy, Perroncito studied the sarcomata of cattle, and claims
to have first observed the micro-organism in 1863. In 1875 he
described it in the Encyclopedia Agraria, and, from the negative
results obtained by inoculation experiments, was led to regard it,
not as the cause, but as a result of the disease.
Rivolta of Turin also claims to have been the first to have
‘discovered the fungus in actinomycosis bovis. As early as 1868
he published a paper on a sarcomatous tumour of the jaw of
an OX,
Hahn of Munich, in 1870, undoubtedly met with the fungus, for
he states that in a case of “‘ wooden-tongue ” he found characteristic
organised structures, which he provisionally described as a species of
mould fungus.
Bollinger was the first to recognise the nature of this disease in
cattle. In 1876 he pointed out that new growths occasionally
occurred on the upper and lower jaws of cattle, which either started
from the alveoli of the back teeth, or from the spongy tissue of the
bone, and by increasing in size loosened the teeth. In their progress
they destroyed bone, muscles, mucous membrane, and skin. After
‘some time they frequently broke down, forming ulcers, abscesses,
and fistule ; but in some cases tumours were formed, which attained
ACTINOMYCOSIS. 419
the size of a child’s head. Bollinger stated that this disease had
been known by various names,—Osteosarkome, Winddorn (Spina
ventosa), Knochenkrebs, Knochenwurm ; in other instances it had been
regarded as bone tuberculosis, or mistaken for a simple chronic
glossitis. Among breeders of cattle and owners of stock in Germany
it had been known under the following names: Ladendruck, Laden-
geschwulst, dicker Backen, Backel, Kinnbeule, Kiefergeschwulst, etc.
Bollinger pointed out that these swellings ‘consisted of several
centres of growth, bound together by connective tissue. They were
often as large as a walnut or a hen’s egg, and of a pale yellow
colour and moist appearance. The cut surface presented yellowish-
white, suppurative foci, while in other cases the growths had a
spongy texture, owing to the formation of lacune or hollow spaces
in a fibrous stroma, which contained a turbid, thick, yellow, caseous
pulp.
Microscopical examination of the tumour showed that it had a
structure like a sarcoma, while the squeezed-out pulp consisted
principally of pus cells, granulation cells, fat granules, and granular
detritus. In addition, there were numerous opaque, pale-yellow, and
coarsely granular bodies of different sizes, which had a mulberry-
like appearance, and were sometimes encrusted with chalk. After
careful examination Bollinger found that these bodies were true
fungi, and he further maintained, from the constancy of their
appearance in all parts of the sarcomatous growth, that they were
not accidental, but of pathogenic significance. This was found to
be the case, not only in fresh preparations, but in old specimens
preserved in the museum. This remarkable form of mycosis was
observed by Bollinger, not only in the upper and lower jaws, but
also in the tongue. It had long been observed that the tongue was
sometimes covered with more or less tubercular growths, scattered
abundantly over the surface of the mucous membrane, mostly the size
of a millet seed or hemp seed, but often reaching the size of a cherry
or walnut, or even larger. In the fresh state these nodules were
greyish-white, and semi-transparent, but they soon became cloudy
or distinctly puriform in the centre; they were surrounded externally
with a connective tissue capsule. If the nodules were situated on
the surface of the tongue, destruction of the mucous membrane very
readily followed, leading to the formation of ulcers. The tongue
also might become affected with an interstitial glossitis, which often,
in spite of the partial atrophy of the muscular fibres, led to a great
enlargement and wood-like hardness of the tongue. On account
of this peculiar character, such a tongue was long known in South
420 INFECTIVE DISEASES.
Germany as Holzzwnge. In other cases the condition was regarded
as “tubercle of the tongue,” “ chronic sarcoma,” “ chronic interstitial
glossitis,” or simply “degeneration of the tongue.”
Bollinger described this disease as occurring in cattle of all ages,
developing itself gradually, and being always incurable. As a rule,
the animals were slaughtered, because the diminished mobility and
enlargement of the tongue interfered with feeding. He also pointed
out that this disease of the tongue was by no means rare, as he had
had no less than six such tongues from different parts of Bavaria in
the space of a year, and he also had been able to prove the existence
of the disease in museum specimens.
On further continuing his researches, Bollinger found the same
fungus in tumours which occurred in the pharynx, larynx, and
the mucous membrane of the stomach. These tumours were very
common in the throat in some parts of North Germany, where as
many as 5 per cent. of the animals had been known to be affected.
The disease frequently occurred in the form of subcutaneous
neoplasms, called Lymphome, Hohzgeschwiilste, Fibrome, Tuberkel,
Tuberkel-scropheln.
This disease also appeared in the form of abscesses, which were
called, in many districts, Schlundbeulen. These growths were found
in the neighbourhood of the parotid gland, the larynx, and pharynx,
and were similar in every respect to the affection of the jaw. They
were described as starting apparently from lymphatic vessels in
these parts. Bollinger discovered the fungus in a case of so-called
fibroid of the second stomach of a cow, a spongy growth nearly the
size of the fist; and he believed that in another case the disease
manifested itself in the form of tubercular ulceration of the
intestines.
Bollinger submitted the fungus to Dr. Harz, a botanist, who
described the fungi as mulberry-like masses from -5 to 1 mm. in
diameter. They appeared to the naked-eye as opaque, white grains,
and when calcified were difficult to recognise. On slight pressure
the tufts of the fungi fell apart into segments of unequal size, each
of which appeared to correspond to an individual fungus. The
latter was described as beginning at the pointed end of the wedge,
with a somewhat cone-shaped basal cell, which, in the absence of a
mycelium, perhaps took its place, and bore a great number of short
linked hyphe. At the ends of the hyphe there were oval, globular,
or elongated club-shaped bodies, the reproductive cells or gonidia.
Cultivation experiments, and inoculation of the tongue of a calf
with liquid containing the micro-organism, failed. Harz proposed
ACTINOMYCOSIS, 42]
to call the fungus, from its ray-like appearance, actinomyces ; but
what the position of the fungus in nature might be, was difficult to
determine. It did not, he believed, belong to the yeast fungi, but to
the mould fungi, and might be compared to Botrytis, Monosporium,
and Polyactis.
Bollinger concluded that there could be no doubt that actino-
mycosis occupied an important position in the pathology of cattle
diseases. As further evidence of the prevalence of the affection, he
remarked that Zippelius of Obernburg had observed in the course of
about ten years’ practice not Jess than 254 cases of lymphoma, in
the neighbourhood of the larynx and pharynx, besides 157 cases
of disease of the jaw; and Bollinger says that he had very little
doubt that the greater part of the former, and very likely all the
cases in the jaw, were due to the fungus which he had discovered.
Tn certain parts of Franconia, according to a communication received
from Professor Frank, these tumours of the throat were extremely
abundant in cattle.
Bollinger’s researches were followed by those of Siedamgrotzky,
and later by a communication from Johne. Johne described the
various forms of the disease which had up to that date been
recognised, including a description of actinomycosis of the ‘bones
of the jaws, of the fauces, of the larynx, of the cesophagus, of the
stomach and intestinal canal, and of the udder. He carried outa
series of experiments, by which it was clearly established that the
disease could be communicated from cattle to cattle. Previously
Bollinger, Harz, Perroncito, Ponfick, Siedamgrotzky, and Johne had
failed, but subsequently by employing fresh material from the
living animal, both Johne and Ponfick succeeded.
Siedamgrotzky not only confirmed Bollinger’s researches, but he
described the presence of the fungus in so-called ‘‘ multiple sarcomas ”
of the mucous membrane of the esophagus. Rabé described the
presence of the fungus in tumours known as Winddorn, and pointed
out that, in at least one case, he considered that the disease had
been carried by the lymphatics. There were eleven subcutaneous
tumours in a row on the face, which were connected by swollen,
rope-like, lymphatic vessels. They appeared to be secondary to a
growth on the nostril, the size of a hen’s egg.
Perroncito described a case of ‘‘sarcoma” of the intestines and
stomach, which proved to be actinomycosis.
Many additional communications were made on the subject of
this disease. Ponfick produced it in the lungs by intravenous
injection, and subsequently three cases occurring naturally in the
422 INFECTIVE DISEASES.
practice of veterinary surgeons were published. They not only
deserve especial mention, but as this form of the disease appears to
be so seldom recognised, they will be given in detail.
Plug described a case in the lungs. The cow had been out of
health for four weeks, did not eat, and had a cough, and two days
previous to the visit had become rapidly worse. Schmidt found
dyspnea with abdominal respiration ; the nostrils were dilated, ‘the
head protruded, and the mouth kept open. There was dulness on
percussion, and crepitation. The animal was killed, and the lungs,
which alone were diseased, were sent to Plug. The pleura on exami-
nation was normal, but beneath it were numbers of miliary
tubercles, many equal in size to a pin’s head. On section the lung
had a granular appearance from the presence of countless numbers
of minute deposits, which all had the appearance of grey tubercles ;
in none was there any central softening. They were present in
enormous numbers around the bronchi, and in the vessels of the
interlobular tissue. Microscopical examination showed, in the middle
of most of these nodules, the presence of greenish-yellow, radiating
bodies, which under a high power appeared to be undoubtedly
actinomycotic granules. In many there were only rudimentary fungi
consisting of four or five clubs; there was only one rosette in each
tubercle. The fungus was surrounded by round cells and fibrous
tissue. Larger nodules resulted from the agglomeration of several
tubercles, or from diffuse infiltration of round cells in the neighbour-
hood of a tubercle.
Hink met with a somewhat similar case. A ten-year-old cow
was slaughtered, and in the middle lobe of the right lung there were
yellowish nodules about the size of a pea, scattered over an area
the size of the palm of the hand. These nodules were not at
first sight distinguishable from ordinary tubercles, but on closer
inspection they appeared to be somewhat different, and could be
easily shelled out from the thickened lung tissue. On making a
section, pus welled up at several points, and contained yellowish,
calcareous particles. These particles, on microscopical examination,
were found to be strongly calcified tufts of the actinomyces embedded
in granulation cells. Addition of hydrochloric acid dissolved the
calcareous matter, but had no action on the fungus.
Pusch described a third case. The lungs of a cow, which had
been killed on suspicion of having pleuro-pneumonia, were sent for
examination. The front lobe of the left lung was collapsed and
firm, the pleura was thickened and opaque; the larger bronchi were
enlarged, filled with pus, and their walls thickened. In the posterior
ACTINOMYCOSIS. 423
lobe of the left lung there was a cavity the size of the fist, which
had been opened, and the contents had, for the most part, escaped ;
what remained was a greyish, purulent liquid, full of yellowish
bodies. By the side of this cavity there was another collection of
pus, the size of a walnut. In the lower part of the second lobe of
the right lung there was a firm, grey tumour, the size of a hen’s
egg, over which the pleura was much thickened. On section this
was cavernous, with similar purulent contents, and yellow grains.
These grains under the microscope proved to be ray-fungi. The wall
of the cavity consisted of dense connective tissue lined with a soft
granulation tissue, bathed in pus. There was no disease of any
other parts in this case, so that it corresponded in this respect with
the two previous ones. Pusch adds that it was difficult to determine
whether the organism had gained access to the lungs by the blood-
vessels, or by the inspired air. In his case he inclined to the latter
view, and concludes by saying that the organism is probably very
common and attached to the most varied objects, from which it
may be conveyed by the air.
Pusch refers in the same paper to an interesting case which
occurred in the practice of Eggeling. The latter had under his care
a cow with extensive paralysis. The spinal cord was compressed
by a compact swelling in the neck, consisting of the nodules of
actinomycosis. There were no manifestations of disease in any other
part of the body.
Prevalence of the Disease.—The author found that the disease
was not generally recognised as a common affection of cattle
in this country, in spite of the interest excited by the work of
Fleming, to whom is due the credit of first recognising a case in
England. In 1887 there was a disease prevailing in Norfolk, and
in the following year outbreaks were investigated by the author
in Essex, Hertfordshire, Cambridgeshire, and Middlesex. In the
Norfolk outbreak the author found on one farm & per cent. of the
beasts affected with the so-called “‘wens” or “sitfasts,” which
proved on microscopical examination to be cases of actinomycosis.
These growths had previously been described in veterinary text-books
as the result of strumous or scrofulous inflammation ; but in all
the specimens of wens received from this country and the colonies,
the author has been able to demonstrate the presence of the ray-
fungus.
A case of pulmonary actinomycosis, with grape-like growths on
the pleura, indicated that wens were not the only manifestation of
this disease, which had keen lost sight of under the designation of
424 INFECTIVE DISEASES.
tuberculosis. Many other cases were examined, and the disease
was shown to be prevalent in this country.
Fic. 178.—From a photograph of a Norfolk steer. There is a growth about the
size of an orange in front of the throat, an example of a so-called ‘‘scrofulous ”
or “strumous” tumour. This growth was associated with a large polypoid
growth in the pharynx which, by interference with deglutition, produced
emaciation (Fig. 180).
In Australia actinomycosis commonly occurs in the form of
tumours of the upper and lower jaw, which were attributed to
“cancer” or to ‘ scrofulous
inflammation.” The diseace
is still commonly known in
Australia as “cancer” and
“lumpy jaw.”
Reports of the prevalence
of actinomycosis in the United
States have been published
by the Board of Live Stock
Commissioners for the State
of Illinois. In their Report
for 1890 several interesting
communications were pub-
Fie. 179.—A Norroik EIF 7 : :
Sa Norrolx HEWER WITH A ished. Mr. Casewell, State
Larcr ‘ WEN” IN THE Parorip REGION. 4 x P :
Veterinarian, investigated an
outbreak of this disease, known also in America as “lumpy jaw,’
ACTINOMYCOSIS. 425
on a farm in Yates City, where there were 80 head of cattle, and
16 were found to be suffering from actinomycosis. Mr. Casewell
Fic. 180.—Photograph of a steer nearly three years old, but about the size of a
yearling. The emaciation and deplorable aspect recall the appearance of
“a, piner ” or ‘‘ waster ” (tuberculosis).
reported that the disease was prevalent in nearly every county in
that State, and that in his opinion it was spreading. In one
instance 109 cases were slaughtered.
Actinomycosis in Relation to Tuberculosis—When we consider
the very high percentage of cases
of tuberculosis which has been
reported in some localities, the im-
portance of differentiating actino-
mycosis from tuberculosis cannot
be over-estimated. The very
great contrast in the appearance
of the micro-organisms in the
two cases renders this a very
easy matter for the pathologist.
But practical veterinarians and
breeders of cattle are liable to
mistake some manifestations of
actinomycosis for tuberculosis.
It is of the greatest im-
portance to bear in mind that
wens or clyers are really not tubercular, but actinomycotic; and
Fic. 181.—Actinomycotic NopuLEs
FROM THE PLEURA.
426 INFECTIVE DISEASES. \
that a condition of the lungs. may occur as the result of actino-
mycosis, which from the naked-eye appearances may be mistaken for
“orapes” or “angleberries.” It will be well also to remember in
connection with the above remarks, that extreme emaciation may
result in actinomycosis, producing a condition which, without a
post-mortem examination, would probably be attributed to tuber-
culosis, the animal being regarded as a “piner” or ‘“ waster.”
If these possible fallacies are taken into account, the excessive per-
centage of tubercular cases so commonly reported will be very
considerably reduced.
There is no evidence to show that the flesh of animals suffering
from actinomycotic tumours is unfit for human consumption. In
very severe cases it is unwholesome, but there is no evidence that
it can produce actinomycosis in man.
MAanirestaTIons oF ACTINOMYCOSIS IN Man.
(I.) Invasion by the Mouth and Pharynx.—The fungus may gain
access through carious teeth, or wounds or fistule of the jaw, and
very possibly by inflammatory processes in the pharynx and tonsils.
The disease attacks the ‘lower jaw most frequently. The tumour
is found in close connection with the bone, or in the sub-maxillary
or sub-mental regions, and also in the pre-tracheal region. It occurs,
though rarely, in the interior of the bone.
In a case described by Israél, which occurred in a woman aged
forty-six, there was a small tumour about the size of a cherry
attached to the external surface of the lower jaw, with an opening
through which a probe could be passed into the bone. The tumour
was incised and scraped away, and a cavity discovered in the bone,
admitting a small sharp-spoon. Later, a further operation was
performed: the periosteum was detached, the cavity of the bone
enlarged, and the contents scraped out, consisting of granulation
tissue, fragments of bone, and the yellowish fungi. At the bottom
of the cavity the fang of the canine tooth was found. No return of
the growth occurred.
The first cases of actinomycosis which were observed in America
were connected with the jaw. In 1884 Dr. Murphy described two
cases at Chicago. The first was that of a woman aged twenty-eight.
Two weeks previously she had suffered from severe toothache, with
swelling in the throat and great pain in swallowing. It disappeared
after poulticing, but she was again attacked with toothache, and a
swelling appeared on the angle of the jaw on the left side. The
ACTINOMYCOSIS. 427
mouth could not be opened without difficulty ; ‘the tonsil was much
enlarged, and pus was set'free on incision. She still suffered with
toothache, and a small swelling now occurred on the left side of the
neck below the jaw. She had several carious teeth. The swelling,
which was about the size of a walnut, was punctured, and a drainage
tube inserted ; a creamy-looking discharge containing yellow granules
continued to escape, but the swelling and induration increased. A
further operation was decided upon. The carious tooth was removed,
and a probe passed into the alveolus showed a communication with
the external wound; the angle of the jaw was chiselled away, and
the alveolus scraped out. Iodoformed gauze was applied, and the
case recovered.
The second case was a man aged eighteen, who had also suffered
with severe toothache and swelling at the angle of the jaw. On
examination a carious tooth was noticed. The swelling was well
marked, and there was fluctuation ; it was as large as a pigeon’s
egg, and situated below the jaw. When punctured, thick creamy
pus escaped containing the fungi; the sinus was scraped out, and in
ten days the wound was healed. Another swelling appeared, and
this was treated as before, and the case recovered.
The peculiar feature of these growths is their apparent migra-
tion. Israél states that in one case a tumour occurred on the alveolar
process, close to carious teeth, and later was close to the edge of
the jaw in the sub-maxillary region. From thence it disappeared,
and a large swelling formed below the hyoid bone, and after this had
been incised and had healed, an abscess formed above the clavicle.
Actinomycotic tumours in this region would sometimes appear to
correspond very closely with wens or clyers in cattle; they may dis-
charge through the skin, and the opening close, or a fistula result ;
but they differ, from their tendency to form burrowing abscesses
instead of recognisable tumours. In this respect they recall chronic
inflammation rather than the sarcoma-like growths in cattle.
Cases in which the upper jaw is attacked are not so frequent as
those in the lower jaw. The progress is usually described as slow,
and there is a tendency for the deep-seated soft parts to be involved,
while in the lower jaw there is a tendency for the tumour to come
to the surface. There may be burrowing suppuration, or small
tumours, which, after a time, fluctuate and form distinct abscesses.
These may involve the skin, discharge their contents, and leave
fistulous openings.
In other cases the disease has been described as extending from
the alvcolar process to the temporal bone, or the base of the skull,
428 INFECTIVE DISEASES.
destroying bones and even reaching the brain; or the growth may
descend by the spinal column, implicating the vertebra, and travel-
ling and pointing in various directions.
(I1.) Invasion by the Respiratory Tract.-_In one recorded case
the disease existed for seven years, was localised to the bronchi
(Bronchitis actinomycotica), and did not extend into the lungs.
The sputum was examined, and contained the characteristic fungus.
If the micro-organisms are inhaled they pass into the bronchioles
and alveoli, and produce proliferation of round cells, which undergo
fatty degeneration. The resulting patches of peri-bronchitis or
pneumonia become yellowish-white; suppuration and hemorrhage
from the capillaries follow, and small cavities result, containing pus
cells, fat granules, blood, and the fungi. In the neighbourhood of
the new growth there is compression of the alveoli, and ultimately the
formation of a dense stratum of connective tissue, separated from the
cavities by a lining of granulation tissue containing the character-
istic fungus. The symptoms are usually obscure; but the sputum
may contain the fungi, which are often visible to the naked eye.
The apices of the lungs are not, as a rule, affected. There is con-
siderable clinical resemblance to chronic phthisis: cough, night-
sweats, pallor, shortness of breath, and hemoptysis are symptoms
common to both. Light may be thrown upon the case by the examina-
tion of the sputum. The presence of the actinomyces will be positive
evidence as to the nature of the disease. The existence of these
symptoms, with absence of tubercle bacilli, would lead to the
suspicion of actinomycosis, even failing the discovery of the fungus
in the sputum.
In the second stage the symptoms are more characteristic. The
disease spreads to neighbouring parts, and pleurisy commonly super-
venes. This extension may involve the peri-pleural tissues. Thus
the disease may follow the pre-vertebral tissues, descend behind the
insertion of the diaphragm, and point as an ordinary psoas or
lumbar abscess; it may perforate the diaphragm and reach the
abdominal cavity. Peritonitis or sub-phrenitic abscess may then
result. In some cases adhesions have formed, and the disease has
extended to the liver or spleen, or other abdominal organs. The
disease may also extend forwards in the direction of the anterior
mediastinum and the pericardium.
The primary affection of the lung becomes of secondary import-
ance. Grave symptoms occur, hectic fever, night-sweats, rigors,
and marked pallor. In the third stage, the disease comes to the
surface, either over the chest, or in the neighbourhood of the dorsal
ACTINOMYCOSIS. 429
or lumbar vertebre ; a swelling appears of a livid colour, and if
punctured no fluid escapes, but if allowed to make its own way to the
surface, the skin gives way, a muco-purulent discharge mixed with
pieces of the growth escapes, and the fungi can readily be recognised.
(III.) Invasion of the Digestive Tract—In a case under Chiari,
death, with general marasmus, took place at the age of thirty-four,
after two years’ illness. The mucous membrane of the intestines was
almost completely covered with whitish patches, raised in the centre,
and covered with yellow and brown granules closely adherent to.
the adjacent tissues. The teeth were carious.
Small nodules about the size of a pea may be found in the sub-
mucous tissue, and in the mucous membrane itself. They soften
and form ulcers with undermined edges, the base reaching the:
muscular layer. They may undergo cicatrisation, but generally
the disease extends through the peritoneum to the abdominal cavity,
and perforates the bladder or the intestines, or makes its way through
the abdominal wall. Symptoms are either absent or not character-
istic. The fungus may sometimes be found in the evacuations, or
by exploratory puncture.
(IV.) Undetermined.—In, addition there are a number of recorded
cases presenting very varied symptoms and anatomical relations, in
which it has not been possible to satisfactorily determine the path
of infection. Delépine has described a most interesting case of an
actinomycotic tumour of the brain.
1
MANIFESTATIONS or ACTINOMYCOSIS IN CATTLE.
(I.) In the Digestive system we find the disease attacking :—
(a) The lips, gums, buccal mucous membrane and palate, and
appearing as nodules, wart-like growths, or ulcers. The nodules
and ulceration of the palate were well shown in a specimen sent to-
the author for examination, under suspicion of being the result of
severe foot and mouth disease.
(6) The upper and lower jaw, where it probably originates in
carious teeth, and extending and invading the neighbouring cavities
and sinuses destroys the tissues with which it comes in contact,
expanding the bones into thin plates or reducing them to the
appearance of pumice-stone.
(c) The tongue, where we see it most commonly in the form of
nodules or wart-like patches under the mucous membrane, with a
special tendency to ulcerate, through the irritation of the teeth. These
nodules may extend into the deep muscles, and often collect in rows.
430 INFECTIVE DISEASES.
more or less parallel to the superficial muscular fibres. Complete
tranverse sections of the tongue, double-stained, readily show this
arrangement, even to the naked eye. Induration of the tongue
results from secondary interstitial glossitis. The author has seen,
in one case only, a tumour embedded in the substance of the tongue
about the size of a small Tangierine orange, and more or less isolated
from any surrounding growth.
(d) The pharynx, where the disease may occur in the form of
polypoid growths producing asphyxia.
Fic. 182.—A Norroik STEER WITH EXTENSIVE ACTINOMYCOTIC ULCERATION OF
THE SKIN OF THE FLANK. D
(II.) In the Respiratory system we may meet with the disease in :—
(a) The nasal cavities, originating primarily there or resulting
from extension of a growth from the lips, or the pharynx.
(b) The larynx and trachea, generally in the form of polypoid
growths, sessile or pedunculated, which arise primarily or occur
secondarily, by extension from the tissues in the neighbourhood. .
(c) The lungs, where the differentiation of the disease is most
important, as neoplasms in the lungs, especially in the early
stages, and nodular growths on the pleura, may be mistaken for
tuberculosis.
The disease is very rarely found in connection with the
Nervous system (II1.), but probably does not so rarely attack the
Reproductive system (1V.).
ACTINOMYCOSIS. 431
(V.) The skin and subcutaneous tissues are a favourite seat of this
disease, producing the so-called wens or clyers socommonly seen in the
fen country. A wen is first recognised as a small tumour, the size
of a marble or walnut, which increases in size sometimes with great
rapidity, and breaks down and discharges its muco-purulent contents
through the inflamed and ulcerated skin ; or it may go on increasing,
and form a large compact growth, the size of a child’s head. These
growths when excised, hardened, and cut, have a characteristic
honeycombed appearance, produced by the interlacing bands of
fibrous tissue, which form a spongy structure, from the interstices
of which the fungus tufts and thick yellowish pus have for the
most part dropped out.
Actinomyces Hominis.—Careful examination of pus from
a case of actinomycosis in man will reveal to the naked eye little
yellowish-white or yellow bodies, which a casual observer might
mistake for grains of iodoform. On collecting some of the discharge
in a test-tube, and holding it between the light and the eye, the tufts
of fungi appeared as brownish or greenish-brown grains, embedded
in a muco-purulent matrix.
On spreading some of the discharge on a glass slip, the largest
tufts of the fungus are found to be about the size of a pin’s head.
They have’ a distinctly sulphur-yellow colour by reflected light, but
appear of a yellowish or greenish-brown tint by transmitted light.
With a sewing needle, or a platinum wire flattened at the end into
a miniature spatula, the grains can be readily picked out of the
discharge, or taken off the dressing, transferred to a clean slide, and
gently covered with a cover-glass. Examined with an inch objective,
they have the appearance of more or less spheroidal masses of a
pale greenish-yellow colour. On removing the preparation from the
microscope, and gently pressing down the cover-glass with the finger,
the grains flatten out like specks of tallow; and on again examining
with the same power they are found to have fallen apart into a
number of irregular and sometimes wedge-shaped fragments of a
faintly brown colour, affording a characteristic appearance. By
preparing another specimen, and covering it with a cover-glass
without completely flattening out the grains, the spherical, oblong
and reniform masses of which the tufts are composed can be
recognised with a }-in. objective as rosettes of clubs. By examining
the peripheral part of a rosette with « 7'y-in., and especially after,
pressing the grains into a thin layer, with or without the addition
of a drop of glycerine, the characteristic clubs are most readily
demonstrated, and the most varied shapes observed by carefully
432 INFECTIVE DISEASES.
examining the form of the individual elements. As in the bovine
fungus, every variation in form is found, from single clubs to clubs
with lateral offshoots, clubs bifid at the extremity, palmate or
fan-shaped groups, and banana-like bunches. In many cases the
clubs are divided by transverse fission into two, three, or more
segments. As a rule, the clubs are irregular in shape, and of about
equal size, while a few are conspicuous by their length. In other
parts of the preparation the clubs are replaced by long slender
forms, which are sometimes transversely divided into a number of
short links. With suitable illumination many clubs are seen to
taper off into slender filaments. In addition there are free filaments,
which are twisted, branched, and sometimes distinctly spirilliform.
Many of the clubs are.composed of layers differing in their refractive
power, and many have the appearance of a central channel. There
are also in the preparation small, highly refractive bodies, fat
granules, granular detritus, round cells, pus cells, and sometimes
blood corpuscles.
The grains differ, as a rule, from those from a bovine source, in
the absence of that sensation of grittiness so often transmitted to the
finger when pressing the cover-glass upon them, and in the slightly
greater tendency of the tufts to retain their compact form. By
teasing the grains in a drop of water on a slide,.and examining
the preparation with a % or a 74 objective, the explanation
of the latter is forthcoming ; for by this process the clubs are
gradually washed away, and a central core remains, which is com-
posed entirely of a dense network of filaments. This can readily be
observed by using a small diaphragm, and it will be found that
the rosettes of clubs are now replaced by tangled masses, having some
resemblance to miniature tufts of cotton-wool. These filaments
constitute the delicate network which is seen in sections stained
by the method of Gram. This can be readily verified by making
a cover-glass preparation of the grains, and staining by that method.
the characters of the fungus can readily be studied by proper
illumination, without staining. The clubs have a faintly greenish
tint, and in form and arrangement are quite characteristic and easily
recognisable. Permanent preparations may be made by mounting
the fungus in glycerine.
Description oF STAINED SPECIMENS.
The fungus may be stained in alcoholic solution of eosin in the
manner to be described for the bovine organism, or in orange-rubin,
DESCRIPTION OF PLATES XV. AND XVI.
Actinomyces.
PLATE XV.
Fic. 1—From a preparation of the grains from an actinomycotic abscess in
a boy; examined in glycerine. The drawing has been made of a com-
plete rosette examined by focussing successively the central and peripheral
portions. Towards the centre the extremities of the clubs are alone
visible; they vary in size, and if pressed upon by the cover-glass give the
appearance of an irregular mosaic. Towards the periphery the clubs are
seen in profile, and their characteristic form recognised. At one part
there are several elongated elements, composed of separatelinks. x 1200.
Fig. 2.—Different forms of clubs from preparations in which the rosettes have
been flattened out by gentle pressure on the cover-glass. x 2500.
(a) Single club. (0) Bifid club. (¢) Club giving rise to four
secondary clubs. (d) Four clubs connected together, recalling
the form of a bunch of bananas. (¢) Mature club with a lateral
bud. (f/f) Apparently a further development of the condition
represented at (e). (g) Club with a lateral bud and transverse
segmentation. (h) Single club with double tranverse segmenta-
tion. (4) Club with oblique segmentation. (j) Collection of
four clubs, one with lateral gemmation, another with oblique
segmentation, (%) Club with lateral buds on both sides, and
cut off square at the extremity. (2) Club with a daughter club
which bears at its extremity two still smaller clubs. (m) Club
divided by transverse segmentation into four distinct elements.
(2) Elongated club composed of several distinct elements. (0) and
(p) Clubs with terminal gemmation. (7) Palmate group of clubs.
(7) Trilobed club. (s) Club with apparently a central channel.
(t) Filament bearing terminally a highly refractiveoval body.
PLATE XVI.
Fie. 1.—From a section of a portion of the growth removed from a boy
during life. The tissue was hardened in alcohol, and cut in celloidin.
The section was stained by Gram’s method and with orange-rubin. x 50.
Fic. 2.—From the same section. “A mass of extremely fine filaments occupies
the central part of the rosette. Many of the filaments have a terminal
enlargement. The marginal part shows a palisade of clubs stained by the
orange-rubin. x 500.
Figs. 3 and 4.—From cover-glass preparations of the fungus teased out of the
new growths produced by inoculation of a calf with pus from a boy
suffering from pulmonary actinomycosis. Stained by Gram’s method and
orange-rubin. The threads are stained blue and the clubs crimson (a)
In the younger clubs the thread can be traced into the interior of the
club (0). In some of the older clubs the central portion takes a yellowish
stain, and in others the protoplasm is not continued as a thread, but is
collected into a spherical or ovoid or pear-shaped mass. In others, again
irregular grains stained blue are scattered throughout the central portion
(Fig. 4). x 1200.
Fig. 5. From a pure-culture on glycerine-agar. (a) branching filaments, (b) a
mass of entangled filaments. Gram’s method. x 1200.
Fie. 6.—From a similar but older cultivation. (a) a filament with spores
(6) chains of spores simulating streptococci. Gram’s method. x 1200.
(nw
INC A ISOM AC ons)
<S = | K\ , if he A
ARNIS Ly. ARS FIC AWS
Fig 4.
"en
On
see —
% b
et We ;
ee 6
Fig6.
Vincent Brooks, Day & Son, Lith.
. ACTINOMYCOSIS. 433
and in either case mounted in glycerine. But although the fungus
can be detected without any staining process, there may sometimes
be doubtful appearances, and then cover-glass preparations should be-
made and stained by the method of Gram with eosin. The filaments
can be readily recognised, and this is of great value, as it forms.
an additional means for the diagnosis of the disease. In combination
with orange-rubin we have a test that is as characteristic and useful
as staining for tubercle bacilli. The discharge, scraping from a
growth, sputum, or the isolated fungus is.squeezed between two.
cover-glasses, which are then slid apart; they are allowed to dry,
passed through the flame in the ordinary manner, and then stained.
The cover-glasses can be cleared in clove-oil, the excess of clove-oil
being removed by gentle pressure between pieces of blotting-paper,.
and then the preparation can be mounted in balsam and rendered per--
manent. On examination of these specimens the masses of filaments.
will be found to be stained blue, and the tissue elements pink. These
filaments vary very much in extent and character in different pre-
parations. In some cases there are masses of short threads, which
are either straight, sinuous, or twisted, and branched. In other
parts the field is occupied by very short, straight, or curved and
sometimes spiral fragments; in others, again, there are comparatively
long strands. On examination with a high power, and with careful
illumination, some filaments will be observed to be moniliform,,.
while others are provided with a terminal oval body. There are
also free spherical, and oval, bodies stained blue, which represent the-
spores of the organism. When orange-rubin is used instead of
eosin, the clubs will be stained and easily recognised. This method
enables one to determine the exact relation of the threads to the
club-shaped bodies ; and this is an interesting point, asit has been
suggested that the threads are not connected with the clubs, but are:
merely an adventitious micro-organism growing in the track of the
ray-fungus. The threads are stained blue and the clubs crimson.
In the younger clubs the protoplasm of the thread can be traced into
the interior of the club. In some of the older clubs the central.
portion takes a yellowish stain, and in others the protoplasm is not
continued as a thread, but is collected into a spherical, ovoid, or
pear-shaped mass. In others again, irregular grains, stained blue, are
scattered throughout the central portion. The sheath of the thread
is stained pink; and the protoplasm, stained blue, fills the sheath, or-
consists of small spherical or irregular grains, giving a distinctly
beaded appearance.
The effect of various reagents should be tried upon the isolated
28
434 INFECTIVE DISEASES.
grains. The grains are picked out of the pus and transferred to
watch-glasses containing strong potash, xylol, and benzol. If
returned to a slide and covered with a cover-glass, the clubs are
found unaltered. Water or weak potash washes away the clubs,
and the filaments become easily distinguished; ether and strong
acids have no effect upon them. Corallin soda, Hanstein’s violet,
and iodine zinc-chloride fail to give any particular reaction,
Hoffman’s blue stains the clubs, but without bringing out any
structural details which could not be observed in the unstained
Specimens.
Actinomyces bovis.—The fungus in cattle may in the same
way be detected with the naked eye in the muco-purulent discharge,
or in a scraping from the cut surface of a growth. The tufts of the
fungus vary in size under different circumstances, from that of a grain
of fine sand to that of a pin’s head. If the pus or scraping be spread |
out on a slide and examined against a dark background, the grains
appear to be white or yellowish-white in colour; but if examined by
transmitted light, they appear distinctly brownish. On pressing the
cover-glass on the slide the grains readily flatten out, being of a
soft, tallowy consistency; or in the process of gently pressing the
cover-glass on the slide with slight lateral movement, a distinct
gritty sensation is transmitted to the finger, owing to the presence
of caleareous matter. On examination with a low power the
fungus will be recognised in the form of irregular patches scattered
over the field, which might readily be regarded as collections of.
granular débris of a brownish or yellowish-brown colour, but on
careful examination they are observed to have a more or less
characteristic appearance. On examining with a higher power,
spherical, ovoid, or reniform bodies are to be seen, which are
either typical rosettes of clubs or granular masses, with here
and there a club-shaped body at the periphery. Pus cells, round
cells, fat granules, and minute spherical bodies may also be
distinguished. If the grains consist of typical rosettes, and be
merely covered with the cover-glass, and examined without being
flattened out between the cover-glass and the slide, they will recall
to mind, on focussing alternately the centre and the periphery,
the appearance of the capitulum of a composite flower. The central
portion appears to consist of spherical forms; these are the ex-
tremities of the component elements, and as we focus the edge of
the rosette these elements are seen laterally, and their characteristic
club-form is readily distinguished, The central portion may be
flattened against the cover-glass, and as the individual clubs vary
DESCRIPTION OF PLATES XVII. AND XVIII.
Actinomycosis Bovis.
PLATE XVII.
Section of an actinomycotic tongue stained by the method of
Gram and with eosin.
Fiq. 1.—This illustrates the appearance which is usually seen under a low
power, when a section is stained by Gram’s method and with eosin. The
central portion of a mass of the fungus is either unstained or tinged with
eosin, while the marginal portion is stained blue. The reverse is seen, as a
rule, in sections from man ; although under a low power the general appear-
ance of sections from these two sources is somewhat similar. x 50.
Fic. 2.—za, b, ¢, d, represent the earliest recognisable forms of the ray fungus
in the interior of leucocytes. In é¢ the club-forms can be recognised. In
f and g there are small stellate groups of clubs. x 500.
Fic. 3.—A part of the section represented in Fig. 1, under a high power. The
marginal line of blue observed under a low power is now recognised as the
result of the stain being limited to the peripherally arranged clubs. At
(a) part of a rosette has undergone calcification ; the clubs are granular,
and have not retained the stain. At (0) and close to it there are the
remains of rosettes in which the process of calcification is almost complete.
x 500. \
PLATE XVIIL
The figures in this plate are taken from sections of a case of
so-called “osteosarcoma,” in which the growth of the fungus was
remarkably luxuriant. The specimens were stained by Plauts’
method.
Fic. 1.—Different forms of clubs in different specimens: x 1200.
(a) Very small club-shaped elements.
(0) A club with transverse segmentation.
(e) A club with lateral daughter clubs.
(@ and ¢) Clubs with terminal offshoots resembling teleutospores.
(f) A club with developing daughter clubs on the left, and on the
right a mature secondary club.
‘ (g) A segmental club with lateral offshoots.
(2) Two clubs undergoing calcification.
Fic. 2.—A very remarkable stellate growth comprised of nine wedge-shaped
collections of clubs radiating from a mass of finely granular material.
x 500. :
Fic, 3.—A rosette undergoing central calcification, and consisting in part of
= extremely elongated clubs resembling paraphyses. Calcareous matter is
also being deposited in the club-shaped structures. x 500.
Fie. 4.—Part of a rosette with continuation of the club-shaped bodies into
transversely segmented branching cells apparently representing short
hyphe. x 500..
Fic. 5.—A rosette from another section in which similar appearances are
observed as in Fig. 4. x 500.
Plate XVII.
ACTINOMYCOSIS BOVIS
Orodkshande fecit. Vincent Brocks,Day & Son, Lith
Plate XVIII.
ACTINOMYCOSIS BOVIS
ACTINOMYCOSIS., . 435
considerably in size, the appearance of an irregular mosaic is thus
produced,
By pressing upon the cover we break up the rosette, and then
the clubs are recognised either singly or in pairs, or attached together
in the form of wedge or fan-shaped segments. Calcareous material,
if present, may readily be demonstrated by the action of acids. It
will be found that on the addition of dilute hydrochloric, nitrie or
acetic acids, the calcareous deposit is dissolved while the clubs are
not affected, and even with the addition of the strongest acids the
only result will be to dissolve out the calcareous matter and clarify
the tufts of the fungus, the form of the clubs being still recognisable.
They are not affected by ether or potash, and thus the effects of
chemical reagents clearly distinguish them from fat erystals or
calcareous particles. By breaking up the growth into small frag-
ments, we may readily study the shape of the individual club-like
elements. By using high-power objectives, and properly arranging
the illumination, various forms will be clearly delineated. In some
cases the club will be found to be bifid at the extremity ; in other
cases there are lateral offshoots or daughter-clubs. Here and there
will be found clubs closely pressed together like a bunch of bananas,
and in other cases the broken-off pieces have a palmate form. By
teasing out the grains in water, and pressing them apart between
the slide and cover-glass, we find that the central portion is com-
posed, as a rule, of a structureless core. More rarely there are the
delicate filaments which are found in cultures and in the fungus from
man,
If the grains are mounted in glycerine, the appearance of the
organism in the fresh state may be preserved.
The granules may be stained by picking them out with needles
and transferring them to a watch-glass containing alcohol, to which a
few drops of concentrated alcoholic solution of eosin have been added.
They remain in the solution until distinctly stained, and they are
then placed on a glass slide in a drop of glycerine.
The muco-pus may be spread out into as thin a film as possible
on a cover-glass, allowed to dry, fixed by warming slightly over
the flame, and stained by the method of Plaut or of Gram. The
, characters of the fungus can be so readily recognised in the perfectly
fresh state, that methods of staining are of secondary importance
in diagnosis, though there are certain minute points which can only
be satisfactorily determined by means of suitable dyes.
436 INFECTIVE DISEASES.
CULTIVATION OF ACTINOMYCES.
Bostrém cultivated actinomyces from five cases in animals, and
from one case in man. In all cases he obtained a similar result.
The fungi were isolated from pus with sterilised needles, and placed
in liquefied nutrient gelatine in which they were teased out, and the
gelatine then spread on glass plates. The growth is stated to have
become visible in a few days. The fungi were isolated from the
plates, crushed between sterilised glass slides and inoculated on the
surface of nutrient agar-agar and blood serum. In this way pure
cultivations were obtained. Nutrient gelatine was not liquefied.
The cultures in blood serum and agar-agar grew best at from 33°
to 37° C. The track of inoculation gradually spread out during
the first two days, having a finely-granular, whitish appearance.
During the next few days small yellowish-red spots appeared in the
centre of the inoculated area, while the edge was apparently com-
posed of fine processes. The yellowish spots continued to increase
for about seven or eight days, and became confluent, and the
periphery also was dotted with yellowish-red points. Finally, there
were also isolated colonies consisting of a yellowish-red centre with
a greyish, downy periphery. The cultivated fungus, if suitably
stained, “corresponded exactly with that found in human and
animal actinomycosis. In the cultures during the first two days
threads were found, with true branchings; later the threads were
divided into shorter pieces or rods, and when the yellowish centres
appeared there were also a number of very short rods and cocci-
like forms. Bostrém also described attenuated club-shaped swell-
ings at the end of the threads. He concluded by saying that
Actinomyces is not one of the mould fungi, the central threads do
not therefore constitute a mycelium; he was inclined to regard it
as a branched Cladothrix, and cultivation seemed to prove this. He
suggested that it might be the Streptothrix Forsteri of Cohn. In
any case he relegated Actinomyces to the fission fungi or bacteria.
In 1888 the author made cultures on glycerine-agar from a case of
human actinomycosis of the thoracic wall. An abscess was opened, the
discharge collected in sterilised tubes, and cultivations prepared with
as little delay as possible. Some of the discharge was spread out on
a sterilised glass slide, and the grains isolated with sterilised needles
and quickly transplanted on the surface of the nutrient medium.
The tubes were placed in the incubator at 37° C., and the result
watched from day to day. For several days there was to the
naked eye no promise of success; but gradually the grains began
ACTINOMYCOSIS. 437
to change, and by the end of a fortnight there was an appreciable
increase in size. Numerous cover-glass preparations were made
from what was originally a single grain, and on examination by the
method of Gram the appearance was very striking. There could be
no doubt as to the increase of the mycelial structure. The dense
masses of filaments covered almost the whole area of the preparation.
In parts less thickly covered there were very numerous oval
bodies, and rod-like segments with terminal enlargements. 'These
“crocus” forms corresponded with the appearances previously de-
scribed as met with in the interior of certain clubs. From this it
would appear that some other condition is necessary for the develop-
ment of the fully formed club, which is the result of the sheath
undergoing some change, possibly mucilaginous, resulting in the
formation of a thick investment of the clubbed mass of protoplasm
at the end of the thread.
These club-shaped bodies represent organs of fructification, rather
than the results of degeneration or death. The difficulty in accept-
ing the view of their being entirely lifeless forms lies in the fact that
the author has observed daughter-clubs growing from the mature
clubs; and, further, in the bovine fungus the author has been
able to trace the stages in the development of a single club to a
completely formed rosette.
In the unstained condition, the clubs are found, on the whole, to
be very regular in their form and arrangement, and by certain
staining methods they can be shown to have a somewhat complex
structure. If we take all the characters into account, and par-
ticularly the minute structure and the relation to each other of the
threads and clubs, we are justified in the opinion that the club in
the early stages is an integral part of the living fungus, and that
these characters bring the fungus into relation with a higher group
of micro-fungi, Basidiomycetes, although the filaments, regarded
by themselves, correspond with the characters of Streptothrix. The
life-history of the micro-organism may be summed up thus :—
The spores sprout into excessively fine, straight, or sinuous, and
sometimes distinctly spirilliform threads, which branch irregularly
and sometimes dichotomously. The extremities of the branches
develop the club-shaped bodies. The clubs are closely packed to-
gether, so that a more or less globular body is formed, with a central
core composed of a dense mass of threads. The threads can be
differentiated by the method of Gram into an external sheath, and
protoplasmic contents. The club-shaped body externally appears to
be mucilaginous, while internally it is continuous with the protoplasm
438 INFECTIVE DISEASES.
of the thread. It is difficult to say what further changes occur in
the club-shaped bodies ; in all probability they represent organs of
fructification. If so, the protoplasm in the interior of the club may
possibly undergo changes leading to the development of spores,
which are ultimately set free; in some cases the terminal segment
of a club is separated by transverse fission in the form of a globular
body, a process resembling the formation of spores by abjunction.
In others, the forms sprouting from the club are suggestive of
teleutospores. There are occasionally long, slender forms, very
different from the ordinary clubs; they possibly represent paraphyses
or abortive elements. In whatever way they may be formed, there
can be little doubt that spores are set free in the vicinity of
a rosette, and give rise to fresh individuals; the ultimate result
recalling, as has been suggested, the appearance of “ fairy rings.”
There can be little doubt that spores and young fungi are taken
up by wandering cells, and conveyed to a distance from the parent
fungus, and thus fresh centres of growth are established.
Appearances of cultwres.—Bostrom, Wolff, Israél, Paltauf, and
others have shown that actinomyces can be cultivated in the
ordinary nutrient media. More recently the author has carried on
a series of cultivations for some years on glycerine agar, gelatine
and milk, broth, bread-paste, and potato, in order to observe the
changes which take place, and to study the variations which he
found in the appearance of sub-cultures. The actinomyces after a
few days on glycerine agar at the temperature of the blood forms
little, white, shining, moist colonies, which may remain stationary, or
increase and coalesce. In a week or ten days, sometimes earlier, and
sometimes after several weeks, the cultures turn a bright yellow colour,
but some remain, though white; others, again, have a tinge of pink,
and others are yellowish-brown (Plate XIX). After atime, a powdery
efflorescence makes its appearance on the surface of the culture,
which may be either yellow or white in colour. The culture may go
on increasing, spreading over the surface of the medium, and retain
its yellow colour, or it may turn black in parts or completely so,
while the agar is coloured brownish-black. Cultures have a peculiar
sour smell; the variations in cultures were all proved, by careful
testing by sub-cultures, to be due to the growth of the actinomyces
under varying conditions of soil, temperature, and the supply of air.
The stage of efflorescence corresponds with the breaking up of the
filaments into masses of cocci, and chains often closely resembling
streptococci. Gelatine is slowly liquefied.
Wolff and Israél cultivated actinomyces on raw and boiled
DESCRIPTION OF PLATE XIX.
Pure-cultivations of Actinomyces.
These tubes were selected from a great number of cultivations
in which there were different appearances. In some instances the
growths had a faint tinge of pink.
Fig. 1.—Pure-cultivation on the surface of potato, showing a luxuriant
sulphur-yellow growth entirely composed of entangled masses of fila-
ments. After three months’ growth,
Fig. 2,—Pure-culture from the same series, on glycerine-agar. In this case
the culture remained perfectly white. The jelly was coloured reddish-
brown. After fifteen months’ growth.
Fic. 3.—Pure-culture on glycerine-agar in which the growth was dark-
brown, in parts black, and the jelly stained dark-brown. After nearly
two years’ growth,
Plate XIX.
Fig 1 Fig 2 Fig 3
PURE-CULTIVATIONS
OF
ACTINOMYCES.
H. Crookshamls,fecit Vincent Brocka, Day & Son, Lith
ACTINOMYCOSIS. 439
eggs, and succeeded, by inoculation in the peritoneal cavity, in
producing the disease in rabbits and guinea-pigs, in the form of
tumours of the peritoneum. Sauvageau and Radais consider that
actinomyces should not be included with bacteria, and have suggested
the name Oospora bovis.
PREPARATION AND EXAMINATION OF TISSUES.
In order to examine the microscopical appearances, the tissues should
be hardened in absolute alcohol and embedded in celloidin. The sections,
when stained, are to be dehydrated as a rule in strong spirit instead of
absolute alcohol, as the latter dissolves the celloidin. If the sections are
very friable, they can be cleared with clove-oil on the slide. By these
means the little fungus tufts, which have a great tendency to fall out of
the sections, may be preserved in situ after passing through the various
staining processes. To cut the sections, we can use either Jung’s micro-
tome, cutting in alcohol, or the freezing microtome. In the latter case,
after the celloidin has hardened, it is necessary to shave off all that
surrounds the piece of tissue. It is placed in water until it sinks, and
then transferred to gum, and frozen and cut in the ordinary way.
Staining Methods.
There are several methods by which the organism can be stained in
the tissues, but it is best to employ for this purpose Gram’s method and
modifications of Plaut’s method.
Gram’s Method.—By Gram’s method the clubs in the bovine disease
are distinctly stained, especially if the sections contain the fungus at a
suitable stage. Use freshly prepared staining solution. A few drops of
aniline-oil are placed in a test-tube, which is filled up with distilled
water, the mouth of the tube closed with the thumb, and the mixture
shaken up thoroughly. An emulsion forms, which is then filtered, until a
perfectly clear solution of aniline-water is obtained. To this is added,
drop by drop, an alcoholic solution of gentian-violet until precipitation
commences. About fifteen to twenty drops in a small capsule of aniline-
water will be sufficient. Sections are floated in this dye for about ten
minutes, then transferred to the iodine-potassic-iodide solution until they
turn brown like a tea-leaf. They are then decolorised in alcohol ; then
stained in a weak alcoholic solution of eosin, dehydrated in strong com-
mercial alcohol, cleared in clove-oil, and mounted in balsam. It will be
found that the clubs are stained blue, and that there is a central area,
which is, as arule, tinged by the eosin. There are various modifications
of the method, and some of them are extremely successful in affording
not only a picture of the fungus, but also the structure of the surrounding
tissue. Very instructive results may be obtained by combining the
method of Gram with Ehrlich’s histological stain. In this case, after
the section has been decolorised in alcohol, it is ready to be transferred
to logwood and treated as described below.
Weigert’s Method.—This also gives very beautiful results. The sections
440 INFECTIVE DISEASES.
are placed for an hour in Wedl’s solution of orseille, which is prepared
as follows :—Add liquid extract of orseille to a mixture of absolute
alcohol 20 parts, strong acetic acid 5 parts, distilled water 40 parts,
until a dark-red liquid results. This must be filtered before use. The
sections are left in this solution for an hour, then just rinsed in alcohol,
and transferred to a solution of gentian-violet. Such sections show the
nuclei of a violet-blue colour, and the peripheral part of the central core
in the larger masses of the fungus also takes a blue colour, while the
club-shaped structures are stained a striking wine-red colour.
Plaut's Method and Modifications.—This is one of the most valuable
methods for staining the clubs. The original method was to float sections
for ten minutes in magenta solution warmed to 45° OC. This solution
consisted of magenta two parts, aniline-oil 3 parts, alcohol of specific
gravity 0:830, 20 parts, distilled water 20 parts (Gibbes). The sections
were then rinsed in water, stained in concentrated alcoholic solution of
picric acid for from five to ten minutes, immersed in water five minutes,
50 per cent. alcohol fifteen minutes, passed through absolute alcohol and
clove-oil, and preserved in Canada balsam. The clubs are stained a
brilliant red and the tissue yellow. Instead of employing the magenta
solution, we now use Ziehl-Neelsen’s solution.
By removing the picric acid in Plaut’s method by prolonged immersion
in alcohol, and then staining with gentian-violet or methylene-blue, a
very successful contrast can be obtained. The most instructive histo-
logical picture can be obtained by first staining with Neelsen’s solution,
removing the stain from the tissue in the way that has been already
described, and then transferring the sections to distilled water, and
subsequently staining with Ehrlich’s histological stain.
Ehrlich’s New Histological Stain,—This is a combination of Ehrlich’s
logwood with orange-rubin. It is of especial value for sections of
actinomycosis, and particularly in combination with carbolised fuchsine.
It is employed in the following way :—The sections must be placed in
alcohol or distilled water, and then in Ehrlich’s logwood for about half
a minute. From this solution they are transferred to distilled water,
washed to remove the excess of stain, and then placed in a large dish of
tap-water, where they are left for half an hour or more, until the
sections turn blue; if preferred, they may be left overnight. They are
then stained for one or two minutes in a solution of rubin, 8, and
orange, and washed again in distilled water to remove the excess. They
must then be dehydrated in alcohol, cleared in clove-oil, and mounted in
balsam.
Preparation of Large Sections.
The method of cutting large sections of organs may be employed
in studying actinomycosis. The value of these sections depends not
‘only upon their affording an instructive picture of the naked-eye
appearances, but they can also be studied with a pocket lens, or under
the microscope with a 4 or }-in. objective. By staining the sections, the
relation of the morbid to the healthy structures is brought out in greater
‘contrast, and thus the topography of the disease can be studied more
‘
DESCRIPTION OF PLATES XX. AND XXI.
Actinomycosis Bovis.
PLATE XX.
Fr¢. 1.—From a section of an actinomycotic tongue stained by the triple
method (Ziehl-Neelsen, logwood and orange-rubin), In this section the
separate centres of growth are clearly shown. Each neoplasm consists of
a fungus system, in which the masses of the fungus, situated more or less
centrally, are surrounded with round cells, epithelioid cells, sometimes
giant cells, and lastly fibrous tissue forming a more or less distinct
capsule. In parts the fungi have fallen out of the section. x 50. |
Fie. 2.—From a section of a “tubercular” nodule from the lungs of a
Norfolk heifer with pulmonary actinomycosis. The nodule is a multiple
growth surrounding a bronchus, and is enclosed by a capsule, in the
vicinity of which the pulmonary alveoli are compressed. It is composed
of a number of separate neoplasms, and each of the latter is composed of
secondary centres of growth resembling the giant cell systems of bacillary
tuberculosis. The new growth is composed of ray-fungi, large multi-
nucleated cells, sometimes distinct giant cells, round cells, epithelioid cells,
and, surrounding them, fibrous tissue. On examination of the same
specimen with a higher power the typical rosettes of clubs are sometimes
surrounded by multinucleated cells, and sometimes small rosettes are
found like tubercle bacilli, in the interior of giant cells. From a pre-
paration stained by Ziehl-Neelsen, logwood, and orange-rubin. x 50.
PLATE XXI.
Fie, 1.—(@) A leucocyte containing the fungus in its earliest recognisable
form. (6) A large multinucleated cell containing the fungus in an early
stage with the club-form already visible. (¢) A leucocyte containing a
small stellate fungus. (d@) A large cell containing clubs arranged in a
small rosette. (¢) A multinucleated cell with clubs arranged in a palmate
form. All the above are drawn from sections of actinomycotic tongues
stained by the triple method, x 500.
Fig. 2.—A giant cell with large vesicular nuclei at the periphery, and in the
centre a fully formed rosette of actinomyces with a smaller growth within
a “daughter” cell. From a section of the tongue of an ox stained by
the triple method. x 500.
Fic, 3.—A very large circular giant cell, with its ring of nuclei at the
periphery, enclosing several isolated tufts of actinomyces. From a section
of a nodule in the lung. Stained by the triple method. x 500.
Fie. 4.—Three rosettes of actinomyces surrounded by a row of large, some-
what angular multinucleated cells. From a section of the tongue of an
ox stained by the triple method. x 430.
Plate XX.
asso
ise
es
Fig 2.
ACTINOMYCOSIS BOVIS
E.M.Crookshank fecit, Vincent Brooks,Day & Son, Lith
Plate XX1.
Fig 1.
a a® & Go
@..@ “e
e sae Bs
ge ; oe
cf ee)
¢ ‘ b &
e ® we G8
iS
NU, 2? @@/ @ Mr Ne y \e
a i | Ge 2 SY!
Mae) gs : .
a = IS ’ @ % ent 4- 8
gt Aas seg c Ns a a
A “(22 2
ACTINOMYCOSIS BOVIS
EM.Grookshanle,fecit
Vincen? Brocles,Day & Son, Lith.
ACTINOMYCOSIS, 441
minutely than by simply observing the cut surfaces of organs or growths.
And, further, it affords a means of rendering permanent many of the
instructive appearances observed at the autopsy, without preserving the
whole structure in the form of a museum specimen. Very satisfactory
results can be obtained with material hardened either in spirit or Miiller’s
fluid. The fresh material is cut with a large, very sharp knife into slices
about a quarter of an inch, or less, in thickness. These slices are placed
between filter-paper in large porcelain dishes, such as are employed for
photographic purposes, and well covered with the hardening solution,
which should be frequently changed. By covering the slice with a small
sheet of glass, which is lightly weighted, any curling or turning up of the
edges is prevented, and the slice not only kept flat, but hardened with
smooth surfaces. Several weeks are required for hardening in Miiller’s
fluid. .The slices, after a short time in water, are placed in gum, and
then frozen and cut ; the slices which are hardened in alcohol are soaked
in water until all trace of the spirit has been removed. A large micro-
tome on the Bruce model is used to freeze and cut the sections. But in
some cases it will be found better to embed the slices in celloidin, and
cut under alcohol with a large microtome of Jung’s pattern. The
sections are carefully removed from the blade of the knife with a large
camel’s-hair brush, and in the case of frozen sections floated in water.
The next process is to float a section out in spirit, and with the
camel’s-hair brush to unfold it and spread it out on a sheet of glass.
The glass with the section is lifted out and examined, and if the section
is sufficiently thin, transferred to the staining solution. In the same
way the section is passed through the various stains, as it should be
- prevented from rolling up or folding in the dye, or it may not be evenly
stained throughout. Modifications of this process will suggest them-
selves, such as pouring off the dye and leaving the section spread out at
the bottom of the dish, and then using the same dish for the next
process. The sections are so easily injured, that it is better, as much as
possible, to avoid handling them. If the sections are only a few inches
in diameter, such as transverse sections of the anterior portion of the
tongue of an ox, they can readily be transferred from dish to dish by
means of a large spatula, made by soldering a piece of sheet German
silver to thick copper wire.
To stain them employ carbolised fuchsine and picric acid, or alum
cochineal, or logwood and orange-rubin. The processes of staining are
precisely the same as with ordinary sections ; but, from their unusual
size, experience and practice are required in their manipulation.
When the section is dehydrated, it is ready to be cleared in clove-oil.
The glass on which it is to be permanently mounted should be selected
without scratches or flaws, and thoroughly cleaned and polished. It is
slipped under the section, which is evenly spread out upon it, and then
lifted out of the dish. The excess of spirit is drained off, the glass
placed on a level surface, and clove-oil poured on the section. It is left
until completely clarified ; the clove-oil, as much as possible, drained off,
and the rest entirely removed by gentle pressure with several thicknesses
442 INFECTIVE DISEASES.
of best filter-paper. In this way several large sections can be cleared at
the same time. But when only one or two sections are dealt with, they
are cleared in clove-oil in a dish, and the mounting glass at this stage
passed underneath them as already described. Another plan, which will
be found of advantage, is as follows :—A piece of clean, thick, filter-paper
rather larger than the section, is slipped underneath it, and then raised
with the section upon it. After allowing the excess of clove-oil to drain
back into the dish, it is carefully laid on the glass with the section down-
wards, and gently pressed down. By taking up a corner, the filter-paper
is peeled off, and the section left behind on the glass. Any creases or
folds are adjusted with needles. After removal of the clove-oil, balsam
is run over the section, and a cover-glass gently and dexterously lowered,
so as to avoid the presence of air-bubbles. The preparations are set aside
to harden in a warm place and on a level surface, and are then ready for
fixing in suitable frames.
Naked-eye Appearances of Large Sections.
In the sections of an actinomycotic tongue it is at once apparent that
the new growth is more or less limited to the periphery of the section.
In parts there are dense clusters of little nodular neoplasms, the fungus
systems, each having a rounded form, and averaging in size that of a
small pea. In other parts small nodules, varying in size from a millet-
seed to a hemp-seed, have a linear arrangement between bundles of
muscular fibres. The appearance is suggestive of an invasion of the
tongue along the lymphatics.
In many of the nodules the largest tufts of the fungus can be seen,
with the naked eye, to occupy a more or less central position. In parts
the muscular fibres are replaced by fibrous tissue.
If now these sections be placed under the microscope, the minute
structure may be examined ; but as it is obvious that still better results.
may be obtained by small sections, any part which it is necessary to
examine with high powers can be selected from a corresponding part of
the growth, and prepared in the ordinary way.
In the case of a “wen,” the whole growth can be excised with the
surrounding tissues, sliced and treated in the way already described, and
sections stained by different methods.
The nature of the growth is at once recognisable as actinomycosis,
from the characteristic honeycombed appearance produced by the trabecule
of fibrous tissue which form a spongy structure, from the loculi of which
the fungus tufts and caseous matter have for the most part dropped out.
In other parts this structure is intact, and the tufts of the fungus can be
detected with the naked eye, and readily recognised with a pocket lens.
A fungus system may be studied more minutely in ordinary sections
of the tongue. Each nodule is composed of the actinomyces surrounded
by round cells and epitheloid cells, and fibrous tissue which often forms
a distinct capsule. In some specimens the fungus is surrounded by a
single row of large multinucleated cells, and in other specimens the
fungus is found in the interior of large oval giant-cells.
ACTINOMYCOSIS. 443
TRANSMISSION OF AcTINOMYCosIS From Man ro roe Lowrr ANIMALS.
Inoculation of cultures has already been referred to. The
author successfully inoculated a calf with material direct from
a living patient.
A calf which had been inoculated in the peritoneal cavity, and
killed seventy days afterwards, presented the following lesions. The
peritoneum of the rumen, in the vicinity of the seat of inoculation,
was studded with hundreds of growths, varying in size from a millet-
seed toa pea. The large growths were composed of several small
ones collected together. On stripping off the peritoneum, and holding
it between the light and the eye, the fungus could be seen with
the naked eye in each individual growth. By incising a growth,
and examining a scraping under the microscope, the characteristic
clubs, and the filaments also, were found to be present. By staining
cover-glass preparations with the method of Gram and orange-rubin,
the appearances were very striking. The clubs were conspicuous on
account of their size, and brilliantly stained, In many, the proto-
plasm of the thread was demonstrated in the interior of the club.
In sections of the peritoneal nodules stained by Gram’s method
the mycelium was found to be present, and the clubs in part
took the stain. With Plaut’s method the clubs were most clearly
demonstrated.
Israél and Johne failed to infect a calf by intravenous injec-
tion, and Ponfick failed to infect dogs, but Israél succeeded with
a rabbit. Israél obtained a small piece of actinomycotic granula-
tion tissue from a peri-pleural abscess in a patient with primary
disease of the lung, and introduced it into the peritoneal cavity.
The rabbit showed no sign of illness, and was killed about ten
weeks afterwards. On examination numbers of tumours were
found in the abdominal cavity, varying in size from a hemp-seed
to a cherry. Larger ones had a somewhat nodular surface with
yellowish points; others were found on the abdominal wall on
the right side. There was « small growth over the psoas muscle,
and one large one attached to an adhesion of the colon. The
growths were on the peritoneum, or attached by longer or shorter
adhesions. Some of the larger tumours showed on section a hollow
space in the centre, which was filled with a pulp, the result of
fatty degeneration of the transplanted tissue, which was sharply
differentiated in colour and consistency from the new growths. The
latter consisted of granulation tissue with abundant formation of
fat granules, blood pigment, aciculay fat crystals, and actinomycotic
444 INFECTIVE DISEASES.
grains. From some of the tumours, radiating processes of a yellowish
colour penetrated into the retro-peritoneal tissue.
Rotter inoculated calves, pigs, guinea-pigs, and rabbits. In one
case he had a positive result. A piece of actinomycotic growth was
introduced into the peritoneal cavity of a rabbit. The animal was
killed six months afterwards, and the piece of tissue which had been
introduced was found to be encapsuled, and around it were twenty
tumours, from the size of a pin’s head to that of a hazel-nut,
containing the ray-fungus.
The author also succeeded in transmitting the disease to a rabbit.
A small quantity of human pus, containing the yellow grains, was
diffused in broth, and injected with a hypodermic syringe into the
abdominal cavity of a rabbit. This rabbit died seventy-nine days
afterwards. On examination several nodules were found, about the
size of a millet-seed, ou the peritoneum of the stomach, in the gastro-
splenic omentum, and on the peritoneum of the diaphragm. There
was a vounded nodule, about the size of a pea, attached to the
stomach. There were adhesions between the intestines, and a
tumour about the size of a marble attached to an adhesion of
the cecum. One of the small nodules was excised and divided, and
the scraping from the interior contained typical rosettes of clubs.
The successful transmission of actinomycosis from man to bovines
suggests intercommunicability, though the negative évidence as to
infection of man from bovines supports the view that the disease
is derived from some source which is common to both species.
TRANSMISSION OF ACTINOMYCOSIS FROM CATTLE TO CATTLE.
Johne was the first to prove that actinomycosis could be trans-
mitted from cattle to cattle, and his results were confirmed and
extended by Ponfick.
A calf was inoculated subcutaneously in the neck and cheek,
in the gum, and the abdominal cavity. The animal died in forty
days after inoculation with development of actinomycosis.
A calf was inoculated in the cheek and abdominal cavity. Death
occurred 114 days after inoculation. In the peritoneal cavity
numerous tumours had formed, and the yellowish grains were ae
to the naked eye in sections of the new growth.
A cow in calf was inoculated in the left posterior quarter of the
udder ; phlegmonous mastitis followed, and subsided leaving a small
induration, which then increased until the inoculated part of the
udder was, in three months, nearly double the normal size, from a
ACTINOMYCOSIS. 445
deposit resembling a fibroma. The cow was slaughtered 133 days
after inoculation. Typical actinomycosis had been produced.
A colt three and a half years old, was inoculated in the jaw
and in the forehead after trephining, and also in the trachea. ‘The
animal died without any result.
Ponfick also conducted a series of experiments which amply
confirmed the results which had been obtained by Johne.
Feeding Experiments.—Repeated experiments, made with masses
of the growth chopped up, or with isolated grains of the fungus, gave
negative results,
Inoculation Experiments.—The growth was inoculated in various
regions of the body. Small particles of the growth from quite fresh
tumours were introduced into the anterior chamber of the eye in
rabbits, with negative results. Rabbits were inoculated in the
peritoneal cavity from an animal recently slaughtered, but they
died of peritonitis. In dogs also the results were negative.
Seven calves were operated on. ‘In five the abdomen was opened
with antiseptic precautions, and in two cases the growth was
introduced by injection. In the latter cases the pieces of tumour
were suspended in salt solution, but the animals died from peritonitis
a few days after the injection. The same result occurred in two out
of the five cases-in which the abdomen was opened. The three
remaining cases gave the following results :—
l.—-Pieces of tissue, about 1:5 cm. long, were taken from the
lower jaw of a recently killed ox. Twelve of these pieces were
introduced into the peritoneal cavity; death occurred after 266
days, from exhaustion and recent lobular pneumonia. At the post-
mortem examination several patches of peritonitis were found, with
encystment of the remains of fragments of the inoculated tissue,
but there was an independent development of several nodules in the
neighbourhood of the stomach and urinary bladder. Examination
of these new formations showed, even to the naked eye, that they
contained yellowish grains, which, on further examination, proved
to be the fungi.
2.—Ten pieces of tumour from the jaw of a cow were introduced
as before; the calf died suddenly sixty days afterwards, during the
injection of fresh pieces of actinomycosis into the jugular vein, At
the post mortem it was found that various adhesions had occurred,
as the result of peritonitis; and in the false membranes there were
sixty-three nodules, varying in size. Microscopical examination
showed that all these nodules consisted of typical actinomycotic new
formation.
446 INFECTIVE DISEASES. .
3.—In this case twelve pieces of growth were introduced into
the abdomen of a calf eight weeks old. Seven days afterwards fresh
pieces were introduced under the skin, in the region of the left lower
jaw. A swelling occurred, which was opened. Pus escaped,
together with the material which had been used for inoculation, in
a state of decomposition. Pieces of tumour were inoculated sub-
cutaneously in the neighbourhood of the right side of the neck,
ninety-nine days after the first experiment. The animal was bled to
death seven months (210 days) after the first experiment. At the
post mortem, peritonitis and adhesions were found, with twenty-one
large and several small nodules in the mesentery, in the false mem-
branes between the viscera, and on and in the serous linings of most
of the abdominal organs. There were also several large and many
small tumours in the subcutaneous and intermuscular tissue, in the
region of the lower jaw and neck on the right side, and numerous
large and small nodules in both lungs, some undergoing softening
in the centre.
Isolated fungi were inoculated in dogs, with negative results,
nothing remaining after 45 to 80 days, except a thick emulsion.
Pieces of tumour were introduced in the submucous and sub-
cutaneous tissue of dogs, but no change occurred after 600 days.
In a calf, inoculation under the gum of the upper jaw showed
what was possibly only the remains of the growth which had been
introduced. At any rate, the experiment was doubtful; but in
a second calf there were numbers of nodules developed around
the points of inoculation in the subcutaneous and muscular tissue
in the neighbourhood of the lower jaw, and in the region of the
neck. The results were observed in these regions in 210 and 110
days respectively.
Inoculation of rabbits and dogs, with isolated fungi, produced
no results after 156, 165, and 170 days. Experiments on dogs,
with grains from a human source, were also unsuccessful. They
were examined after 470 days, and there was no sign of any
result.
After intravenous injection, positive results were discovered in
the only calf which survived this operation, on bleeding the animal
to death 110 days afterwards. Numerous nodules (27) were dis-
covered in the parenchyma of both lungs, without suppuration.
Two dogs were injected in the jugular vein with isolated fungi
mixed with 60 grammes of salt solution. Examined after 45 and
80 days respectively, the lungs and all the other organs were found
to be free from growths.
ACTINOMYCOSIS. 447
Ponfick thus summarised these experiments with the bovine
fungus :—
1. Rabbits and dogs possess a marked immunity from actino-
mycosis, whether pieces of tumour or isolated grains are administered
by feeding, or by inoculation in the serous cavities, in the subcutaneous
or submucous tissue, or by intravenous injection.
2. The most common subject of actinomycosis, the cow, possesses
a not less marked susceptibility to the artificial production of the
disease. By feeding, an infection was not obtained, probably
because the mucous membrane had not been injured; but by
inoculation, on the contrary, an independent growth of fresh
neoplasms was produced in the subcutaneous and intermuscular
tissues, occasionally in the submucous tissue, and in a decided
manner in the abdominal cavity. Clear evidence of this growth
is obtained in some cases within a month, or after three or
four months.
3. By intravenous injection, also, it is possible in a few months
to cause typical new growths in the lungs.
Mapura DIsEAsE.
Mycetoma, or Madura foot, is a chronic local disease, attacking
chiefly the hands and feet, and having considerable resemblance to
actinomycosis. It is a disease of tropical climates, and is commonly
known as the “ fungus-foot disease” of India. A small tumour forms
on the hand or foot, which after a year or two suppurates and
bursts, leaving one or more sinuses, from which peculiar black
particles, or white or pinkish roe-like bodies, are discharged.
The disease in the foot may commence in the big toe and spread
upwards, involving the leg as far as the knee, and even the thigh.
In a typical case the foot is enlarged and painful, and later there
are several sinuses from which a purulent and blood-stained discharge
can be expressed, containing the characteristic particles.
According to Bocarro all early growths are superficial. Dissec-
tion of the growths during an operation, or sections made through
the diseased tissues after excision or amputation, show that the
disease begins generally in the loose cellular tissue, generally the
subcutaneous tissue, and thence extends along the sheath of muscles
and tendons to other soft tissues, and finally the bones.
There are several facts in connection with the causation of the
disease, which are of great interest when it is compared to actino-
inycosis in cattle. Bocarro states that the disease originates in
448 INFECTIVE DISEASES.
wounds, sores and pricks of thorns, and that the points of the
thorns of the Acacia Arabica have been found embedded in the
diseased parts. The disease is common among the agricultural
class, and in 90 per cent. of the cases observed in the Hyderabad
Civil Hospital it occurred in the hands and feet.
Vandyke Carter was the first to point out the resemblance to
actinomycosis, and he believed that the two varieties of the disease,
the black and the white, were the result of the growth of a mycelial
fungus, Chionyphe Carteri.
Kanthack pointed out, that if portions of the growth were
placed in ether or chloroform, and afterwards well washed in
Fic. 183.—Part or Human Foor with Mapura DIskAse.
caustic potash, small rounded bodies were left, which showed rays
under the microscope closely resembling the appearances in actino-
mycosis, and that the reaction of the fungus to staining reagents
was identical with actinomyces. Hewlett examined sections from
the disease in the foot, and also found filaments and clubs.
Boyce and Surveyor examined a number of cases, and care-
fully studied the fungus in the black and white varieties of
the disease. In the black variety the particles were found to
vary greatly in size, from that of a grain of gunpowder to that
of a marble. If the particles were boiled for from a few minutes
to one hour in concentrated caustic potash, and then transferred
to distilled water, the brown colouring matter was removed and a
MADURA DISEASE. 449
mycelial fungus could be seen. If tissue, containing particles, was
washed for about a minute in eww de Javel and then stained, the
colouring matter was removed, and the relation of the fungus to
the tissue could be observed. This fungus consisted of large radiating
and branched hyphe, like those of a species of aspergillus, or mucor.
Sections of the white, fish-roe bodies showed, usually in the centre,
numerous, small, reniform, deeply-stained masses, surrounded by
a radiated zone, with the presence of dwarfed club-like elements
resembling actinomyces.
The author suggested that possibly the presence of the coarser
septate mycelium of the black variety might be attributed to a
mixed infection.
Vincent in Algiers succeeded in cultivating the micro-organism,
and showed that it was a new species of streptothrix.
Streptothrix madurz.—Vincent found that the streptothrix
at first grows scantily in the ordinary culture media, and in such
liquids as Cohn’s solution. In broth, at the end of about a fortnight,
there is a limited growth composed of small, round, greyish masses,
and in sub-cultures the growth becomes more abundant.
The little colonies float in the clear liquid when the tube is
shaken, and subside to the bottom when the liquid is at rest, while
some adhere to the sides of the tube. They may be very small, or
attain the size of a pea; after two months they acquire.a reddish
tinge. Later, on the surface of the liquid, there is a white efflores-
cence composed of spores.
The streptothrix grows well in slightly acid infusions of hay or
straw, the proportion of hay to water being 15 grammes to the
litre. Vegetable infusions, made with carrots, turnips, and potatoes
(20 grammes to 1,000 of water), are suitable media, The
streptothrix grows at the temperature of the room, but best at 37° C.
and with free access to air. Inoculated in the depth of gelatine there
is a scanty growth in the track of the needle and on the surface ;
but it grows best in a nutrient medium, composed of infusion of
hay or potato 100 cc., gelatine 9 grammes, glycerine 4 grammes,
and grape-sugar 4 grammes. Glelatine is not liquefied.
Ordinary nutrient agar is not a very favourable medium, but
on glycerine-agar with grape-sugar there is an abundant growth of
circular, projecting, shining colonies, slightly yellowish-white, which
later become pink or bright red. When the colonies are numerous.
they remain small, but isolated colonies increase rapidly ; they are
depressed in the centre or umbilicated, and the central part remains
white while the periphery becomes red. Later, the culture loses its.
29
450 INFECTIVE DISEASES,
colour and becomes a dull white. The growth is very adherent to
the surface of the jelly, and so tough that it is almost horny. The
Streptothrix can be cultivated in milk, which it slowly pepto-
nises. It cannot be grown on blood serum or egg. On potato, on
.the fifth day at 37° C., there are little prominent colonies, which
slowly increase. After a month the growth acquires a pale-rose
colour, which gradually increases and changes to orange or dark
red. The colour is most intense on acid potato, and after a time
an efflorescence, or whitish dust, appears on some of the colonies,
consisting of spores. The growth is hard and friable.
Rabbits, mice, guinea-pigs, and a cat, were inoculated sub-
cutaneously with particles from the disease, or with cultures; but
only a local nodule was produced in each case, which, after a slight
increase, subsided. Nocard confirmed these results. Intra-peritoneal
and intravenous and subcutaneous inoculations in guinea-pigs, rabbits,
pigeons, fowls, dogs, and sheep were negative; and no trace could
be found of the cultures in any animal subsequently killed and
examined. According to Bocarro, though fresh particles from the
disease inoculated in rabbits and dogs gave negative results, inocu-
lation of cultures in guinea-pigs, rabbits, monkeys, and rats, pro-
‘duced a local tumour of slow growth, which, on section, had the
character of the inoculated material.
From these experiments we must conclude that the disease has
not been shown to be transmissible to the lower animals, by either
inoculation of the diseased tissue, or by cultures of the Streptothrix ;
and the exact relation of both the Streptothrix and the mycelial
fungus to the disease must be considered an open question.
CHAPTER XXXI.
GLANDERS.
GLANDERS is a specific inoculable disease of equines, characterised
by the formation of nodules and suppurating tumours, with which
characteristic bacilli are associated. The disease has been known
from very early times. It is described in books of the sixteenth
century and in very early treatises on farriery. It attacks horses,
asses, and mules. Man and many of the lower animals ean be
readily inoculated, but cattle and swine have an immunity. The
disease is especially prevalent in towns, or wherever horses may be
crowded together without those sanitary arrangements which are
so much attended to in private stables; and in large establishments
‘fresh horses are being constantly introduced to replace others, and
thus the opportunities for the importation of the disease are multi-
plied. The disease varies in its virulence. It may occur in a form
which proves fatal in a few days, or it may exist for months or
years without attracting notice, and yet be capable of being trans-
mitted to other animals. The term ‘“‘farcy” is applied when the
disease manifests itself in the form of tumours in the skin.
Glanders in the horse most commonly produces ulceration of the
nostrils and enlargement of the glands. It commences in the form
of nodules of the mucous membrane resembling miliary tubercles,
and, like them, consisting of a collection of round cells. They
suppurate and coalesce, forming irregular ulcers and raised, congested
nodules. The lymphatic glands become enlarged and suppurate,
and the disease extends to the respiratory organs. In the lungs, in
the early stage, the disease is readily mistaken for tuberculosis.
The nodules suppurate, and cavities are formed, but they do not
tend to caseate. A glairy or muco-purulent discharge from the
nostrils should lead to very careful inspection, with every possible
precaution ; and it will probably be easy to detect the ulceration of
the nostrils. In other cases there may be slight discharge from the
451
452 INFECTIVE DISEASES.
nostrils and swelling of the glands, and nothing more will be visibl
until a post mortem examination has been made.
Glanders in man is found amongst those whose duties brin
them into contact with diseased horses, such as stable-men, cavalr
soldiers, and veterinary surgeons; and it is generally the resul
of accidental inoculation of a wound. An abscess develops, followe
by metastatic purulent infiltration of the lungs, liver, spleen, an
bones. There may be cedematous swelling of the face and ulcer
in the nostrils. The joints may become swollen and painful.
The nodules consist of fibrous tissue and cells, with a tendency t
suppuration. In the lungs the disease spreads by the lymphatics
Fic. 184.—BactLut or GLANDERS; section of a glanders nodule, x 700 (FLUGGE).
The infiltrated patches are necrosed in the centre, which is sur-
rounded by leucocytes and fibrous tissue.
The bacilli were discovered by Loffler and Schiitz in 1882. They
are found in the discharge from the nostrils, in the pus, and in the
nodules of animals artificially infected.
Bacillus mallei._Rods, with rounded ends, shorter and
thicker than the tubercle bacillus, occurring singly, or in pairs,
and sometimes in filaments. The protoplasm in the rod is broken
up in stained preparations, as in the tubercle bacillus. They stain
with the watery aniline dyes, and intensely so with alkaline methylene
blue or Neelsen’s solution ; they are non-motile and aerobic ; spore
formation has been described. They can be cultivated on the usual
media, especially on glycerine- agar and on potato; but they will not
grow in infusions of hay, straw, or stable manure. On the surface
of glycerine-agar a colourless, transparent growth occurs on either
GLANDERS. 453
side of the track of the needle; on glycerine-agar with milk a
whitish layer develops, which gradually changes in colour from
amber yellow to a reddish-brown. On blood serum the growth is
transparent and yellowish ; on potato it is much more characteristic.
After two or three days at the temperature of the blood, a film
develops in the vicinity of the inoculated area, which is honey-like,
transparent and yellow; the transparency disappears, and in a
week the cultures have become reddish-brown. (Plate II., Fig. 6.)
The disease has been communicated to man by accidental
inoculation with a hypodermic syringe which had been used for
inoculating cultures. Horses, asses, cats, goats, field mice, and
Fic. 185.—Suction OF A BRANCH OF THE PULMONARY ARTERY SHOWING
GLANDERS BACILLI PENETRATING THE WALL (HaAMILToy).
guinea-pigs, can all be infected with pure cultures; rabbits, sheep,
and dogs are slightly susceptible; cattle, swine, and white mice
have an immunity. In the guinea-pig a swelling occurs at the seat
of inoculation, followed by the formation of a prominent tumour,
developing into an abscess. The skin becomes involved, and an ulcer
with indurated margin results. The lymphatic glands also become
implicated, and general; infection follows, extending frequently to
the testicles or ovaries, and death results after several weeks. In
rabbits there is generally only a local abscess induced, which termi-
nates in a quickly-healing sore. Mice die in three or four days
from general infection; glanders nodules are found in the liver and
454 INFECTIVE DISEASES.
spleen, closely resembling miliary tubercles. Léffler recommends
inoculation of guinea-pigs as the most reliable method of diagnosis.
Sub-cultures of the bacillus rapidly lose their virulence. The
toxic products have been already described (p. 48). Mallein can
be prepared from a culture on sterilised potato by extracting
with glycerine and water, or from a culture of the bacillus in
broth. A virulent culture is obtained from a glandered horse,
or from a guinea-pig inoculated with fresh virus. Sub-cultures are
prepared in glycerine broth, and incubated at 37° C. for a month
or six weeks. If the cultures are found to be pure they are sterilised
in the usual way, in the steam steriliser, and by filtering: through
porcelain a pale, amber-coloured liquid is obtained. To test for
glanders, a few drops (24 cc.) are injected underneath the skin, in the
middle of the side of the neck. In healthy horses there is no re-
action, or a very slight elevation of temperature. In glandered horses
there is marked rise of temperature (101° to 105°), considerable local
swelling at the seat of inoculation, and signs of general disturbance,
while the glandered tumours become more swollen and painful. The
temperature of the horse to be injected should be taken night and
morning for two or three days before the operation; and in horses
suffering from febrile disturbance the test should be delayed.
Thomassen made a number of experiments on horses suffering from
pleurisy, bronchial catarrh, strangles, and other diseases, without
any reaction, except in a glandered horse, used as a control experi-
ment. Hunting and M‘Fadyean, in this country, have made most
careful observations and experiments, and there is no doubt that
mallein is a very valuable aid in the diagnosis of glanders. In
many cases the reaction has been obtained in horses, and the
existence of glanders has been discovered only after a most searching
post-mortem examination, With this means of diagnosis it is now
possible to determine exactly which are the infected animals in a
stable where the disease has broken out. The animals can then
be slaughtered, and the disease prevented from spreading.
Stamping-out System.—Whatever may be the stage of the
disease or the extent or variety of it, isolation ought to be carried
out in its most complete form—namely, slaughter. The disease
‘might be completely stamped out, if it were not for the difficult
question of compensation. It can undoubtedly be checked by the
existing laws.
In 1869 glanders was included in the list of contagious diseases,
and the provisions with regard to giving notice of the disease, the
regulation of movement, or exposure, and disinfection, were applied to
GLANDERS. 455
horses suffering from glanders. Under an earlier Act it was made.
an offence to expose glandered horses in markets or on commons.
In 1878 power to slaughter was incorporated in the Animals Order.
(1) Where a person having a horse, ass, or mule in his possession, or
under his charge, gives notice to a constable that the horse, ass, or mule is
‘ affected with glanders, or any person is convicted of an offence against
the Act of 1878 by reason of his having failed to give such a notice, then,
if at any time thereafter it appears to the Local Authority, on a special
report of a Veterinary Inspector, that the horse, ass, or mule is affected
with glanders, and the horse, ass, or mule is alive at the end of fourteen days
after the receipt by the Local Authority of that special report, the Local
Authority may serve on the owner of the horse, ass, or mule a notice in
writing requiring him to slaughter it, or to permit them to slaughter it,
within a time specified in the notice.
(2) If in any case the owner fails to comply with the requisition of
the notice of the Local Authority, he shall be deemed guilty of an offence
against the Act of 1878, unless he shows to the satisfaction of the court
of summary jurisdiction before which he is charged that the horse, ass,
or mule, is not affected with glanders, or that the slaughter thereof is.
for any reason unnecessary or inexpedient.
(3) The provisions of this Article may be put in force, from time to
time, as often as occasion requires, in relation to the same horse, ass, or
mule, on a further special report as aforesaid.
In the order of 1892 it was provided that glanders should include
farcy, and power was given to compel slaughter, and to compensate
by payment of half the value of a diseased animal, not exceeding £20,
and full value in the case of healthy animals. Owing to objections
urged against the payment of compensation, another order was
passed, which came into operation at the end of 1894; the order to
slaughter being amended as under :—
(1) A Local Authority may if they think fit cause to be slaughtered .
any diseased horse, ass, or mule, provided that if the owner of the horse,
ass, or mule gives notice in writing to the Local Authority, or their
inspector or other officer, that he objects to the horse, ass, or mule being
slaughtered, it shall not be lawful for the Local Authority to cause that
horse, ass, or mule to be slaughtered except with the further special
_ authority of the Board of Agriculture first obtained.
(2) A Local Authority may, if they think fit, cause to be slaughtered
any suspected horse, ass, or mule, having previously obtained the consent
of the owner thereof.
(3) The Local Authority shall out of the local rate pay compensation
as follows for any horse, ass, or mule slaughtered under this article :—
(a) Where the horse, ass, or mule was diseased the compensation shall
456 INFECTIVE DISEASES.
be such sum as the Local Authority think expedient, being a
minimum in the case of a horse of two pounds, and in the case of
an ass or mule of ten shillings ; provided that in no case shall the
amount of compensation, if above the said minimum, exceed one-
fourth of the value of the animal immediately before it became
diseased,
(b) In every other case the compensation shall be the value of the
horse, ass, or mule immediately before it was slaughtered.
CHAPTER XXXII.
TETANUS, RABIES, AND LOUPING-ILL.
TETANUS.
TETANUS is a communicable disease of man and the lower animals,
characterised by spasmodic contraction of the muscles. It is cow-
monly the result of an injury, and occurs especially after wounds
produced by means of splinters of wood or contaminated with earth
or dust, and may follow after surgical operations.
Carle and Rattone first showed, in 1884, that the disease could
be communicated from man to other animals. Rabbits inoculated
with pus from a case in man developed tetanus, and from these
rabbits the disease was conveyed to others. Nicolaier, the following
year, found that mice and rabbits inoculated with earth often
contracted tetanus, and that the pus which formed at the seat of
inoculation contained, amongst other organisms, characteristic bacilli.
Pure cultures were first obtained by Kitasato.
Bacillus of Tetanus.—Slender, straight rods, and filamentous
forms. Spore formation takes place at the end of a bacillus, giving
it a drumstick appearance. They stain with aniline dyes, but best
with Neelsen’s solution, or by Gram’s method. By the Ziehl-Neelsen
method and methylene-blue, the spores can be stained red, in
contrast to the bacilli, which are stained blue. ‘The bacilli are
anaerobic, liquefy gelatine, and_are slightly motile. They can be
grown at the ordinary temperature, but most readily at the tem-
perature of the blood, in an atmosphere of hydrogen, especially after
the addition of 1 or 2 per cent. of grape sugar to the nutrient
medium. The young colonies on plate-cultivations somewhat
resemble those of Bacillus subtilis. They have an opaque centre,
and are surrounded by fine rays, extending in all directions like
thistle-down. In the depth of gelatine a ray-like growth occurs
in the lower part of the track of the needle. The gelatine is lique-
fied very slowly, and gas is given off. The cultures possess a
457
458 INFECTIVE DISEASES.
characteristic odour. In slightly alkaline broth, and peptone with
alkaline reaction, in an atmosphere of hydrogen, the gas formed
will be sufficient to break the flask if it is sealed up. Kitasato
obtained his cultures from pus, by taking advantage of the resist-
ance of the spores to high temperatures. By raising cultures.
to 80° C. for three-quarters of an hour, the micrococci and bacilli
in the mixed culture were destroyed, while the spores of the
tetanus bacillus retained their vitality, and then sub-cultures were
obtained in a pure state.: The spores are said to be killed by
exposure to steam for five minutes. A 5 per
cent. solution of carbolic acid with ‘5 per cent.
of hydrochloric, will destroy the spores in two:
hours. Kitasato and Weyl obtained tetanin
from pure cultures of the bacillus, Brieger
having previously obtained it from impure
cultures. A tetano-toxin, indol and phenol, and
butyric acid are also found. Brieger and
Frankel attribute the pathogenic properties to
a tox-albumin. These products have been de-
scribed more fully in a previous chapter (p. 41).
A pure-culture produces tetanus in a mouse
in twenty-four hours, and rabbits, guinea-pigs,
and rats can also be infected. No pus forms
at the seat of inoculation, as after inoculation
of earth, but the spasms commence in the
muscles nearest to the seat of inoculation. A
trace of a broth culture will kill a guinea-pig,
the symptoms developing in three days.
Kitasato succeeded in making animals im-
mune to tetanus, and subsequently the discovery
was made that the blood in immune animals
Fic. 186.—Purn-cur. Will produce immunity in other animals, the
TURE or Terranus explanation being that the toxic principle of
Bacinir IN GRAPE-
SUGAR GELATINE. wee a
Four days old, tetanus antitoxin; and if equal parts of the
the tetanus bacillus induces the formation of
(FRANKEL AND serum of an immune animal, and a fatal dose of
PEEIEFER)) tetano-toxin, are together injected into a healthy
guinea-pig, tetanus will not follow, showing that the virus has
been neutralised. Tizzoni and Cattani found that blood. from an
immunised dog was not only capable of completely neutralising the
toxic power of filtered cultures, but that the injection of the blood-
serum produced immunity in otherwise susceptible animals, except
DESCRIPTION OF PLATE XXII.
Bacillus tetani.
Fig. 1.—From a cover-glass preparation of a pure-cultivation of the tetanus
bacillus in broth; stained with Neelsen’s carbolised fuchsine. - 1200.
Lamplight illumination,
Fic. 2.—From a cover-glass preparation from the same source ; stained with
Neelsen’s solution and methylene blue. x 1200. Lamplight illumination.
Plate XXIL.
BACILLUS TETANI
EM.Crookshamle fect Vincent Brocks,Day & Son, Lith.
RABIES. 459
guinea-pigs or rabbits. This antitoxin is possibly secreted by special
glands, such as the thymus and thyroid; and, according to Brieger
and others, extracts of the thymus gland are antitoxic to other
toxins, as well as the tetanus toxin.
These experiments have resulted in the employment of tetanus
antitoxin as a therapeutic agent (p. 63).
RaBiEs.
Rabies, or hydrophobia, is a disease like tetanus, the symptoms
being produced by a virus acting upou the nervous system. The
specific virus appears to originate in dogs, wolves, and jackals ; and
by wounds inflicted by rabid animals, or by inoculation, the disease
may be communicated to man, cattle, sheep, deer, cats, rabbits, and
swine. d
Among the early symptoms observed in the dog are sulkiness
and restlessness, depraved appetite, and irritability. A peculiar
expression of the countenance may be noticed, with twitchings of the
eyes and face, and the animal's attention appears to be fixed upon
some imaginary object. A rabid dog is constantly trying to drink,
and at times howls or barks in a peculiar tone, whilst the breathing
becomes very irregular. On the fourth day, or later, death follows.
After death the glands are enlarged and congested, the tonsils are
inflamed, and the vessels of the epiglottis injected. In some cases
there is inflammation of the lungs, and the stomach may contain a
mass of straw, hair, and horse-dung. The membranes of the brain
and the spinal cord may be also congested. All these symptoms
may be present, or some only, or they may be entirely absent; and
it is partly for this reason that Pasteur’s researches have been of
such enormous value. Very little was known of the experimental
production of rabies until Pasteur commenced his investigations ; and
the test, which can be applied by inoculating rabbits, is invaluable
as a means of diagnosing rabies with absolute certainty. Dogs
suffer from symptoms simulating those of rabies ; and formerly, when
human beings were bitten, it was impossible in some cases to
determine whether the dog had been suffering from rabies or not.
We are indebted to Pasteur for the only reliable test which can
be applied ; and we are now in a position, when a human being is
bitten by a dog supposed to be, but not really, rabid, to remove
all cause for the anxiety which would otherwise remain for months
and even years,
In man the period of incubation lasts from eight days to
A460 INFECTIVE DISEASES.
several months, and in rare cases a much longer period. The wound
from a bite may have healed, and may again become inflamed,
and symptoms follow, owing to the poison affecting the brain, spinal
cord, or the peripheral nerves. In the dog the disease appears in
two forms—raging madness and dumb madness; and the identity of
the virus in the dog and in man is shown by the fact that virus
from man can produce both forms of the disease in the dog. The
virus may be obtained by inoculation of the saliva of a rabid dog;
but this method is uncertain, as other micro-organisms are present.
Pasteur endeavoured to obtain it in a pure state, and was able
to demonstrate that the spinal cord, the brain, and the nerves
contain the virus. It was also found that by direct inoculation of
the nervous system the most certain results followed.
Bacteria in Rabies.—Cocci have been described in connection
with hydrophobia by Fol, Babés, and Dowdeswell. The cocci, it is
said, were observed in sections of the spinal cord of rabid dogs. The
descriptions given by different observers vary considerably, and there
is not any particular coccus constantly associated with the disease.
Nor have any of the bacteria been isolated from the diseased animal,
which were alleged to be present in stained preparations. By many,
hydrophobia is believed to be due to the presence of a micro-
organism, but at present the nature of the contagium is unknown.
Protective Inoculation.—Pasteur found that a dog inoculated
under the dura-mater with virus from the spinal cord of a rabid animal
will develop rabies, as a rule, within eighteen days. By trephining
rabbits and inoculating the virus, and by, in the same way,
transmitting the virus from rabbit to rabbit, the incubation period
gradually shortens, until it is reduced to six or seven days. The virus
has then reached its maximum virulence in the rabbit, and is “ fixed.”
Pasteur then studied the possibility of producing immunity. The
medulla of a rabbit, containing the virulent virus, was suspended in
a glass bottle over caustic potash at a temperature of 25°C. If a
number of spinal cords were thus treated, and examined froin day
to day, it was found that they gradually lost their virulence,
becoming completely inert in from sixteen to twenty days. A series
of cords was thus obtained with diminishing virulence ; by injecting
subcutaneously an infusion of rabid spinal cord crushed in broth,
and beginning with an inert cord on the first day and using the
next in the series on the second day, and so on till a fresh spinal
cord could be injected, it was found that dogs were rendered
insusceptible to the strongest virus, administered by inoculation or
by exposing them to the bites of rabid dogs. Dogs have usually
RABIES, 461
an incubation period of several weeks, and Pasteur conceived that it
would be possible to anticipate the symptoms, which would naturally
follow in a dog which had been bitten or inoculated, by giving
them a mild form of hydrophobia by the injection of attenuated
virus of short incubation period. These experiments showed that
it was possible to do this, and the outcome was the introduction of
a system of protective inoculation in the human subject. Pasteur
succeeded in giving immunity from hydrophobia to about fifty dogs
of every age and breed.
"In 1885 Joseph Meister, a boy nine years of age, bitten badly
by a mad dog upon the hands, legs, and thighs, was brought to
Pasteur. At a post-mortem examination of the dog, its stomach
was found full of bits of hay, straw, and wood, and it had been
unquestionably rabid. On July 6th, sixty hours after Meister had
been bitten, a syringe full of marrow from a rabbit which had died
on June 21st, and therefore fifteen days old, was injected beneath
the skin over the right hypochondriac region. The next morning
Meister was inoculated with a spinal cord fourteen days old, and so
on every day, till on the sixteenth a cord only one day old was used.
So many injections, however, need not have been given, as it was
subsequently found that the spinal marrows injected during the
first five days were inert when tested on rabbits. The marrows of
the next five days showed an ascending scale of virulency, until, on
the last two days of the treatment, Meister had been inoculated
with a virus so virulent that it was capable of causing hydrophobia
in dogs after ten days’ incubation. Meister remained completely
free from hydrophobia. From that time to the present day many
thousands of patients have been treated in Paris by slightly modified
methods, and it is very generally believed that a real prophylactic
agent has been discovered.
Pasteur suggested that the rabic virus might consist of two
distinct substances—a living virus capable of developing in the
nervous system, and a secondary product which, in sufficient pro-
portions, might have the property of hindering the development
of the living virus. The nature of this living virus is quite
unknown.
According to a return of the inoculations at the Pasteur In-
stitute, the total number of persons treated in 1895 was 1,523, of
whom five died. In three cases the symptoms of hydrophobia
ocenrred within a fortnight of the last inoculation. If these three
cases are omitted, the number of persons treated is reduced to 1,520
and the deaths to two. The results are shown in the following table,
462 INFECTIVE DISEASES.
for the nine years previous to 1895, during which Pasteur’s method
has been in operation :—
Year, vpemons | Saat | Rate ot
inoculated.
1886... teas 2,671 25 0-94
1887... me eis 1,770 14 0-79
1888... zi “ee 1,622 9 0°55
1889... ston hie 1,830 ve 0:38
1890... sis aie 1,540 5 0:32
1891... ue wis 1,559 4 0:25
1892... ae he 1,790 4 0°22
SOS Ste ad nse 1,648 6 0:36
1894... = 1,387 vi 0°50
1895... — ~_ 1,520 2 0-13
Of the 1,520 persons treated, 156 were bitten on the head or
face, 829 upon the hands, and 535 on other parts of the body; 122
were bitten by animals experimentally proved to be rabid, 949 by
animals declared by veterinary certificate to be rabid, and 449 by
animals supposed to be rabid.
Babés at Bucharest inoculated 300 persons in one year, and
claimed to have reduced the mortality to -4 per cent.
Stamping-out System.—There is every reason to believe that
rabies could be stamped out in England in six months, if a general
order for muzzling were enforced, and all ownerless dogs were
slaughtered. It is the ownerless cur, the vagrant dog, which is
mainly responsible for the spread of rabies ; and if a general muzzling
order cannot be put into force, it would undoubtedly check the disease
if all dogs were compelled to wear a collar with the name and address
of the owner, and all dogs without owners were destroyed.
LovuPING-ILL.
Louping-ill is regarded by some as an infective disease. It is
a disease of sheep, characterised by symptoms due to an affection of
the central nervous system. The symptoms consist in contractions
LOUPING-ILL. 463
of the muscles of the head and limbs, loss of co-ordination and finally
complete loss of the power of movement. The name is derived from
the peculiar jumping movements in the early stage.
Klein and M‘Fadyean independently investigated this disease.
Klein found bacteria in the cerebral fluid. No micro-organisms
were found in the blood. Special attention was drawn to a
bacterium which was found by Klein in six out of seventeen cases,
and to a micrococcus by M‘Fadyean.
Bacteria in Louping-Ill.—Klein’s Bacterium. Oval cocci and
rods ‘6 tol w in length, ‘2 to -3 win breadth. Colonies in gelatine,
yellowish by reflected light, are brown by transmitted light. On the
surface of gelatine the bacteria form a film, which is crenated at
the edge, and thick in the middle, at first grey and later yellowish.
In the depth of gelatine a filament forms, composed of closely
aggregated minute greyish colonies, and a prominent yellow growth
occurs on the free surface. The gelatine is not liquefied. The
bacteria grow in milk, and broth becomes turbid in two days, and
there is a copious flocculent greyish precipitate.
Injection of broth cultures subcutaneously in rabbits, guinea-
pigs, and mice, produced no result, except local swelling at the seat
of inoculation, which subsided without causing any constitutional
symptoms. The results were equally negative when the cultures
were injected subcutaneously in lambs.
M‘Fadyean’s Micrococcus.—Cocci *3 in diameter. The colonies
are flat, nearly circular, and have a smooth edge. In old colonies
the centre appears as a dark spot. Gelatine is rapidly liquefied,
and a nearly colourless precipitate forms at the bottom of the
tube. Cultures on the surface of agar have a faint yellow tinge.
On potato the colour is deeper but the growth not so well marked.
Milk is coagulated. In broth there is an abundant growth render-
ing the liquid turbid and depositing a white precipitate. The micro-
cocci stain by Gram’s method. Inoculated in rabbits or guinea-pigs,
they produce suppuration ; in horses and bovines, an inflammatory
swelling results without suppuration. They produce abscesses in
sheep and lambs. The cocci were isolated from abscesses in lambs
suffering from louping-ill. Though it is admitted that louping-ill
belongs to the class of infective diseases, there can be no doubt from
these experiments that the nature of the contagium is unknown.
CHAPTER XXXIII.
FOOT-ROT.
SHEEP are subject to several diseases which are classed as foot-rot.
There is one form, known as contagious foot-rot, which prevails in
certain localities, especially on wet land. Brown describes the
disease as primarily a disease of the skin, inducing exfoliation of
the cuticle, and exudation of fluid containing epithelial scales. The
inflammation extends to the membrane of the foot, leading to exfolia-
tion of the hoof, and development of epithelial scales, which form an
imperfect horny layer on the diseased membrane. In one outbreak
investigated by Brown, the disease in the early stage was confined
to the skin between the digits of the fore-feet; the surface was
red, tumid and pulpy, and white purulent matter existed on the
inflamed parts. Later, the hoof grew to an extraordinary length,
fungoid growths made their appearance, developing into foot-rot in
an advanced form.
In France, according to Fleming, the contagious character of
this disease has long been recognised. So long ago as’ 1805 Pictet
imported 200 half-bred merino sheep, some of which were infected
with foot-rot, and placed them with 200 healthy sheep, and in a
short time all the sheep were infected. Several years later Favre
and Sorillon carried out investigations which conclusively proved
the infective nature of the virus. Among other experiments it
was found that when healthy sheep were inoculated in the feet
with virus from diseased sheep, the disease was communicated.
Contagious foot-root may be spread by healthy sheep receiving
the virus from infected sheep in fairs and markets. Ships, railway-
trucks, and carts in which diseased sheep have been conveyed, unless
subsequently thoroughly disinfected, may be the means of trans-
mitting the virus to healthy sheep. Healthy sheep turned into
pastures quite recently occupied by diseased sheep may be inoculated
from the discharges from the feet of the diseased sheep, which
contaminate the grass and soil.
464
FOOT-ROT. 465.
Law believed that the disease arose from an undiscovered micro-
organism, which was probably present in infected pastures. Others
in this country have disputed the contagious character of the disease,
and considered that the same conditions of the pasture which
produced the disease in a flock would produce it again in imported
animals, which would account for the apparent contagiousness.
Brown, in order to test the question of contagion, placed infected
sheep in a pen, the bottom of which was covered with straw which
was not removed while the experiments were in process. Three
healthy sheep, from a locality where foot-rot was unknown, were
placed with the infected sheep. At the end of ten days the feet
of the sound sheep were still healthy. Subsequently two of the
sheep were inoculated, and it was found that the virus introduced
Fic. 187.—Foot or SHEEP SHOWING Fic. 188.—SEcTION THROUGH THE
Disease or Horn (Brown). Foot SHOWING A CRACK EX-
TENDING THROUGH THE WALL.
subcutaneously in the vicinity of the foot produced the incipient
stage of the disease. On making further experiments the contagious
nature of one form of foot-rot was established, but it appears that
the contagious property is only developed after a long period of
exposure, and under certain conditions. On a dry soil the disease
will quickly subside, but on moist land the contagious form of foot-
rot may be communicated by simple contact, in from six weeks to
three months.
From these and other experiments Brown has drawn the
following conclusions :—
1. That so far as the evidence goes it justifies the statement that
foot-rot is a contagious disease ; the infective matter being active
when brought into contact with the skin between the claws, or
30
INFECTIVE DISEASES.
466
*(NMOUG) SMVID
GHL NAXMLAG NING JO ASVASICG,
10 WYO GHONVATY—
O6T “OTH
‘(NMOUG) SHLMOWS
CIOONO,T HIIM CHYTAOO ANVUANAYL ONILALOTS—"GgT
“SI,
rt
FOOT-ROT. 467
when introduced into the system by inoculation, and probably when
taken in by the mouth from contaminated pastures.
2. That it cannot be produced by long-continued exposure to
undrained, moist soils, with an abundant, coarse and wet herbage.
3. That animals exposed to these conditions for many months,
and resisting entirely the influences named above, contract foot-rot
Fie. 191.—Distortion oF Hoor In AN ADVANCED Form oF Foot-rot (Brown).
in from fourteen to twenty-one days on being placed among sheep
suffering from the disease.
4, That sheep affected with foot-rot may improve, and from
time to time become worse; and finally may recover and present a
perfectly healthy condition of foot, notwithstanding that they have
been kept the whole period under the conditions which induced the
disease.
468 INFECTIVE DISEASES.
5. That the contagium of foot-rot remains for some time in
the system (ten to twenty days and longer) without any indication
of disease appearing in the skin between the claws. An infected
sheep may therefore escape detection even by an expert, and may
introduce foot-rot into a sound flock.
One attack does not confer immunity. The disease has been
known to recur in sheep which have only recently recovered from
an outbreak.
The disease is no’, communicable to other animals, including
man, The flesh is harmless, but as in severe cases the sheep are
emaciated, the carcass is in consequence of little, if of any, value
as food.
The nature of the contagium is unknown.
Stamping-out System.—Sheep should not be allowed to be
moved from an infected district except for slaughter. When sheep
are purchased to add to’ the stock on a farm, they should be
isolated for two or three weeks, and carefully watched before they
are allowed to mix with other sheep. If this disease is detected
in a flock, every animal should be carefully examined, and any
suspicious as well as any diseased sheep should be completely isolated
from the rest. Fleming recommends that those which have been
in contact, though still apparently quite healthy, should as a pre-
cautionary measure, be made to pass through a trough containing
a solution of chloride of lime to a “depth of about four inches.
The solution is made by adding one or two pounds of chloride of
lime to two buckets of rain-water. If the trough is placed at the
entrance to the sheepfold, the sheep will be compelled to traverse
it at least twice a day. It is also recommended, when diseased
sheep are treated by this plan, that after recovery, all manure
in the fold should be removed and destroyed, and the soil dug up
to the depth of six inches, or lime freely spread over the surface.
Troughs and hurdles must be thoroughly disinfected, and buildings
freely ventilated after similar treatment. No locality can be con-
sidered free from suspicion until one or two months have elapsed
since the recovery of the last case.
CHAPTER XXXIV.
FOUL-BROOD—INFECTIOUS DISEASE OF BEES IN ITALY—PEBRINE—
FLACHERIE—INFECTIOUS DISEASE OF CATERPILLARS.
FouL-BRooD IN BEEs.
FovL-Broop is a contagious disease attacking bees and especially the
larve. The larve rapidly lose their healthy appearance, die and
decompose, turning into a coffee-coloured mass. The cells of the
honeycomb are mapped out by the dark-brown cappings of the cells
Fic. 192.—DisEasED ComB (Cowan).
containing the diseased larv. The decomposition is associated with
the production of an unpleasant odour, which can be detected at
some distance from an infected hive. The disease has been known
from very early times. Preuss investigated it microscopically, and
attributed it to ‘ micrococci.” These were really the spores of a
bacillus, which was first observed by Cohn. Later, Cheshire and
469
470 INFECTIVE DISEASES.
Cheyne investigated foul-brood, isolated the bacillus, and fully
described its morphological and biological characters in nutrient
media. By removing the cap of
one of the diseased cells, the de-
composing larva can be withdrawn,
and cover-glass preparations will
reveal delicate bacilli and large oval
spores. The bacilli can be readily
isolated and cultivated in nutrient
gelatine.
Bacillus alvei (Cheshire and
Cheyne). Rods varying in size,
and forming large oval spores.
Fic. 193.—Spores or BaciLius 4
on They are motile and possess flagella.
Cultivated in nutrient gelatine in
test-tubes, a delicate ramifying growth appears on the surface,
and irregular whitish masses arise along the needle track. Pro-
cesses shoot out from these masses, and
extend through the gelatine for long
distances. They are thickened at points
in their course, and are clubbed at the
ends. The gelatine is gradually liquefied,
and the bacilli form a loose, white, floccu-
lent deposit at the bottom of the tube.
The liquid in the tube becomes yellowish
in colour after a time, and gives off an
odour of stale, but not ammoniacal, urine.
On the surface of nutrient gelatine the
bacilli grow out in chains of rods in single
file, or of rows of several side by side.
The processes which are formed tend to
curve, and at a short distance from the
track of the needle form a distinct circle,
from which another process grows out,
and a fresh circle is developed. The
gelatine in the vicinity of the bacilli
gradually liquefies, and channels are
formed in the gelatine in which the Fic 194.—Pure-cuntorE IN
bacilli move backwards and forwards. HAV GATED Ents
: ae (CHESHIRE AND CHEYNE).
On nutrient agar-agar a whitish layer .
develops, consisting of bacilli arranged side by side, which in a
few days are replaced by rows of spores similarly arranged. On
INFECTIOUS DISEASE OF BEES IN ITALY. 471
potatoes they form a dryish, yellow layer, and in milk a tremulous
jelly. A cultivation of the bacillus in milk, sprayed over a honey-
comb containing a healthy brood of bee larve, produced foul-brood.
Adult bees fed on material containing bacilli became infected.
Inoculation of mice and rabbits with the bacillus gave doubtful
results.
Stamping-out System.—The infected bees, combs, frames and
quilts must be destroyed, and the hives thoroughly disinfected, as
this is the only way in which the resistant spores can be got rid of.
Cowan believes that if foul-brood were under Government inspection
and infected hives were destroyed, the disease could be stamped out.
Fic. 195.—-CULTIVATION ON THE SURFACE OF GELATINE, x 80
(CHESHIRE AND CHEYNE).
Infectious Disrase or Bers in ITAty.
In Italy bees are subject to another infectious malady, and
Canestrini has found bacilli in the bees and in the larve, which are
believed to be the cause of the malady.
Bacillus of Infectious Disease of Bees (Canestrini).—Rods
2» in width and 4 to 6 yw in length, occurring singly, in pairs
and in chains, sometimes capsulated. They are motile, and spore-
formation is present. They liquefy gelatine, colouring the liquid
pink and forming a white deposit. On agar they form a white
growth, and on potato /a claret-coloured layer. Cultures are said
to be capable of producing the disease in bees and larve,
PEBRINE.
The silkworm disease known in France as pébrine is characterised
by the appearance of black patches on the skin of the worms. It
was investigated by Cornalia, Nageli, and Pasteur. Pasteur’s
472 INFECTIVE DISEASES.
prophylactic measures have been described in another chapter
{p 7).
Panhistophyton ovatum. (Lebert. Nosema bombycis, Micro-
coccus ovatus, Corpuscles du ver-d-soie).—Shining oval cocci, 2 to 3 uw
long, 2 » wide, singly and in pairs, or masses; or rods, 2°5 mw thick,
and twice as long. They multiply by subdivision. They were
experimentally proved to be the cause of pébrine, gattine, maladie
des corpuscles or Flecksucht; and were discovered in the organs of
diseased silkworms, as well as in the pupe, moths, and eggs.
Metchnikoff believes that these micro-organisms are not bacteria,
but psorosperms. /
FLACHERIE.
Silkworms are also subject to a very destructive disease known
as flacherie, flaccidezza, maladie de morts blancs. The worms cease
feeding, die and become a putrid mass. The disease is dependent
upon bad hygienic conditions, and is very infectious. The cause
has not been determined with certainty, but it has been attributed
to a streptococcus. ‘
Streptococcus bombycis (Mikrozyma bombycis, Béchamp).—
Oval cocci 15 » diam., singly, in pairs, and in chains. They are
said to be present in dust from infected localities.
‘
Disease oF CATERPILLARS.
Forbes has described an infectious disease of the larve of a
caterpillar (Pieris rape). Cocci which were found singly and in
masses, were regarded as the cause of the malady.
PART III.
SYSTEMATIC AND DESCRIPTIVE.
473
CHAPTER XXXV.
CLASSIFICATION AND DESCRIPTION OF SPECIES.
In reviewing the history of the various classifications which have
from time to time been proposed, we shall see that the gradual
improvements in the means of studying such minute objects,
the methods of cultivating them artificially, and of studying their
chemistry and physiology, and the ever-increasing revelations of the —
microscope, have resulted in establishing these microscopic objects as
members of the vegetable kingdom, ranking among the lowest forms.
of fungi, but with regard to the division into genera and species we
are still in a position of doubt and uncertainty.
Miiller, in 1773, was the first to suggest a classification. He
established two genera, Monas and Vibrio, and grouped them with
the Infusoria. In 1824 Bory de Saint Vincent also attempted
a classification; but it was not until Ehrenberg in 1838, and
Dujardin in 1841, worked at the subject, that a scientific distinction
of species was attempted.
Ehrenberg described four genera :—
I. Bacterium . . filaments straight, rigid.
II. Vibrio . filaments snake-like, flexible.
TIL. Spirillum . filaments spiral, rigid.
IV. Spirocheta . . filaments spiral, flexible.
Dujardin united Spirillum and Spirocheta, and classed them
thus :—
I. Bacterium . filaments rigid, vacillating.
II. Vibrio ’ . filaments flexible, undulatory.
III. Spirillum . . filaments spiral, rotatory.
Bacteria were still considered as Infusoria, but in 1852 Perty
maintained that some of the smallest living organisms belonged to
the animal and others to the vegetable kingdom, and that Vibrios
without question belonged to the latter. In 1853 Robin pointed out
475
476 SYSTEMATIC.
the affinity of the Bacteria and Vibrios to Leptothrix; and Davaine,
in 1859, insisted that the Vibrios were vegetables, and were in
fact allied to the Alge.
Since that time a flood of light has poured in upon this subject
through the writings of Hoffmann, Pasteur, Cohn, Rabenhorst,
Hallier, Billroth, Warming, Nageli, Magnin, Marchand, Sternberg,
Van Tieghem, Koch, Fliigge, De Bary, Zopf, Biichner, Hueppe,
Marshall Hall, and many others who have studied the morphology,
life-history, and classification of bacteria.
Of all these writers we are most indebted to Cohn, not only
on account of his researches, which extended over many years, but
also for his system of classification, which has since been almost
universally adopted.
Tn his first. classification, published in 1872, Cohn considered the
Bacteria as a distinct group belonging to the Alge, and divisible
into four tribes, including six genera :—
I. Sphzrobacteria . globules (Micrococcus).
II. Microbacteria . short rods (Bacterium).
III. Desmobacteria long rods (Bacillus and Vibrio).
IV. Spirobacteria spirals (Sphirocheta and Spirillum).
Cohn noted, in spite of placing them with the Alge, that the
absence of chlorophyll connected the Bacteria to Fungi, and we find
Naegeli subsequently adopting this view, and employing the term
Schizomycetes or Fission-fungi.
Billroth, in 1874, disputed the division into species, and con-
sidered that all the forms described by Cohn were but developmental
forms of one micro-organism, Coccobacteria septica. In the following
year Cohn answered the criticism of Billroth, and produced a second
classification, in which he still maintained that distinct genera and
species existed. Cohn considered the genera to be distinguished by
definite differences in shape, which were adhered to throughout life,
while some special feature, as a difference in size or physiological
action, or some minute difference in form, determined the various
species.
The second classification of Cohn (1875) only differed from the
first in that, instead of keeping the bacteria as a separate group, he
placed them, from their close relationship with the Phycochromace,
under a new group, the Schizophytes, and added the genera Lepto-
thrix, Beggiatoa, Crenothrix, Sarcina, Ascococcus, Streptococcus,
Myconostoc, and Streptothrix.
Fliigge retained the term Schizomycetes, and divided them thus :—
CELLS ROUND OR OVOID.
CELLS CYLINDRICAL.
CLASSIFICATION AND DESCRIPTION OF SPECIES.
Lal
477
SCHIZOMYCETES (Fiiteen’s OriIcInAL CLASSIFICATION).
/Isolated, or in chains,
or united in amor-
phous. gelatinous
families
Forming gelatinous
families of definite }
form.
Colonies solid, filled
with cells
Colonies, with simple
layer of cells at the
In large numbers,
in irregular
colonies 7 :
In small but defi-
nite numbers, in
regular groups.
Micrococcus.
Ascococcus.
Sarcina.
. Clathrocystis.
q \ periphery
(Short. Isolated, or in smail
heaps loosely united,
or in irregular gela-
tinous families
Short,
distinctly
jointed
(Straight Long,
filaments. | not dis- | Thin
tinctly {| Thick
( Without jointed.
- ramifica-<
tions.
( Threads
isolated, Short rigid
inter- Wavy, or
Ls .
laced, or inepialb Long flexile . .
in bun-
Long. « dies.
Pseudo-ramifications .
\ \ Threads in roundish gelatinous masses
. Bacterium.
. Bacillus.
. Leptothria.
. Beggiatoa,
. Spirochete
(vibrio).
Spiriilum.
Streptothria.
Clathrothrix.
Myconostoc.
478 SYSTEMATIC.
‘
The belief is nevertheless rapidly gaining ground that the lowest
forms of vegetable life cannot be divided by a hard-and-fast line
into a series with chlorophyll (Algz), and a series without it (Fungi),
and the tendency now is to solve the difference of opinion between
Cohn and Nigeli by following the example of Sachs, and amalga-
mating the two series into one group, the Thallophytes.
Researches by competent observers have more recently clearly
demonstrated that several micro-organisms in their life cycle exhibit
successively the shapes characteristic of the orders of Cohn.
This doctrine of pleomorphism, now widely accepted, was dis-
tinctly foreshadowed in a publication by Lister in 1873, though
this memoir contained certain conclusions which have since been
abandoned. Lister described forms of cocci, bacteria, bacilli, and
streptothrix in milk, which he regarded as phases of the same
micro-organism, Bacterium lactis. As a result of his observations,
Lister remarks that “any classification of bacteria hitherto made
from that of Ehrenberg to that of Cohn based upon absolute mor-
phological characters is entirely untrustworthy.” To Lankester,
however, belongs the credit of having definitely and precisely formu-
lated this doctrine. In a paper, also published in 1873, Lankester
observed that the series of form-phases which he had discovered in
the case of a peach-coloured bacterium led him to suppose that
the natural species of these plants were “ within the proper limits
protean, and that the existence of true species of bacteria must be
characterised, not by the simple form-features used by Cohn, but by
the ensemble of their morphological and physiological properties as
exhibited in their complete life-histories.” Lankester inferred that
these phase-forms were genetically connected, in that they all pos-
sessed the common characteristic of a special pigment, bacterio-
purpurin. These conclusions were vigorously opposed by Cohn, and
doubt still remains in the minds of some as to whether the different
forms are really only stages in the life-history of a single species.
Nevertheless the theory of pleomorphism has steadily gained ground
ever since.
Cienkowski and Neelsen worked out the different forms assumed
by the bacillus of blue milk ; Zopf has in a similar manner investi-
gated Cladothrix, Beggiatoa, and Crenothrix, and traced out various
forms (Fig. 196) ; Van Tieghem has investigated Bacillus amylobacter
with a similar result; Hauser has described bacillar, spirillar,
spirulinar, and various other forms in the Proteus mirabilis and
Proteus vulgaris. These facts obviously shake the very foundation
of Cohn’s classification, and we are left without possessing a sound
CLASSIFICATION AND DESCRIPTION OF SPECIES. 479
basis for classification into genera or species. The mode of repro-
duction is not sufficiently known to afford a better means for
distinction than the other morphological appearances taken alone;
nor can we depend upon physiological action, which is held by
many to vary with the change of form, according to altered
surroundings.
Pe
ce
Pe
yy)
45
ri?
Pes,
\
V4
F
Fic. 196.
Ciaporarix DicHotoma.—A. Branched Schizomycete : («) Vibrio-form ; (b) Spiril-
lum-form [slightly magnified]. B. Screw-form at the ends: (a) Spirillum-
form ; (b) Vibrio-form. C. Very long Spirocheta-form. D. Branch fragment,
at one end Spirillum-form, at the other Vibrio-form. E. Screw-form:
(a) Continuous ; (6) Composed of rods; and (c) Cocci. F. Spirochzta-form :
(a) Continuous; (6) Composed of long rods; (c) Short rods; and (d) Cocci
(Zopf).
Zopf, who has warmly supported the pleomorphism of bacteria,
has suggested as a result of his investigations a division of the
Schizomycetes, Spaltpilze, or Fission-fungi, into the following four
groups :—
480 SYSTEMATIC.
ZOPF’S CLASSIFICATION.
Group I. Coccacr#.—Possessing (so far as our knowledge at present
reaches) only cocci, and thread-forms resulting from the juxtaposition of
cocci. The fission occurs in one or more directions.
Genus I. Streptococcus (Chain-cocci).—Division in one or more direc-
tions, Individual cocci remain united together to form chains.
Genus IL. Merismopedia (Plate-cocci),—Divisions in two directions,
forming lamelle or plates.
Genus III. Sarcina (Packet-cocci).—Division in three directions, form-
ing colonies in cubes or packets.
Genus IV. Micrococcus (Mass-cocci).—Division in one direction, cocci
after division remain aggregated in irregular clusters, or singly, or
in pairs or in chains of three or four elements.
Genus V. Ascococcus (Pellicle-cocci).—Like micrococcus, but the cocci
grow in characteristic gelatinous pellicles.
Group II. Bacrerrace#.—Possessing mostly cocci, rods (straight
or bent), and thread-forms (straight or spiral). The first may be absent,
and the last possess no distinction between base and apex. Division (as
far as is known) occurs only in one direction.
Genus I. Bacterium.—Cocci and rods, or only rods, which are joined
together to form threads. Spore- formation absent or unknown.
Genus II. Spirillum.—Threads screw-form, made up of rods (long or
short) only, or of rods and cocci. Spore-formation absent or
unknown.
Genus III. Leuconostoc.—Cocci and rods. Spore-formation present in
cocci.
Genus IV. Bacillus——Cocci and rods, or rods only, forming straight or
twisted threads. Spore-formation present either in rods or cocci.
Genus V. Vibrio—Threads screw-form in long or short links. Spore-
formation present.
Genus VI. Clostridium.—Same as bacillus, but spore-formation takes
place in characteristically enlarged rods.
Grovr III. LeprorricuEa#.—Possessing cocci, rods, and thread-
forms (which show a distinction between base and apex). The last
straight or spiral.
Genus I. Crenothriz.—Threads articulated ; cells sulphurless ; habitat
water.
Genus II. Beggiatoa.—Threads unarticulated; cells with sulphur
granules ; habitat water.
Genus III. Phragmidiothrix.—Threads jointless ; successive subdivision
of cells is continuous ; cells sulphurless ; habitat water.
Genus IV. Leptothrix.—Threads articulated or unarticulated ; successive
subdivisions of cells not continuous ; cells sulphurless.
Grove IV. CxiaporricH#E.—Possessing cocci, rods, threads, and
spirals. Thread-forms provided with false branchings.
Genus :—Cladothrix.
CLASSIFICATION AND DESCRIPTION OF SPECIES. 481
Zopf, however, does not assert that all the fission-fungi exhibit
this pleomorphism, nor does he pretend that his classification will
include all the micro-organisms described. Cohn, on the other
hand, was ready to admit that all the forms described by him were
not truly independent species. De Bary, Hueppe, Baumgarten, and
Fliigge have expressed other views with regard to the classification
of bacteria.
De Bary divides them into two great groups—-bacteria which
form endospores, and bacteria which form arthrospores. This
affords but little practical assistance, though regarded by
botanists, from a scientific standpoint, as a step in the right
direction.
Hueppe, acknowledging that the fructification must eventually
be made the basis for classification, suggests an arrangement for
provisional use in which this view is introduced (p. 482).
It has already been mentioned that the production of arthrospores
is only established in a very few species. Therefore, we are
hardly justified in assuming that all bacteria, the spore-formation
of which is quite unknown, are to be included with those in which
this kind of fructification has been observed, and consequently to
distinguish genera on the same grounds may be considered, to say
the least, somewhat premature. In Baumgarten’s classification the
genus bacterium is dispensed with, and the genera divided into two
groups, the monomorphic and the pleomorphic.
Groupe I.—MonomorpPHic.
Genera.—Coccus.
Bacillus.
Spirillum.
Grover II.—PLeEomMoRPHIC.
Genera.—Spirulina.
Leptothrix.
Cladothrix.
Fliigge also, in his revised classification, includes the genus
bacterium in the genus bacillus. The new classification differs
also from the original one in the grouping together of the different
species according to the character and behaviour of the colonies
in nutrient gelatine. The abolition, in Fliigge’s and Baumgarten’s
classification, of the genus bacterium is no doubt owing to confusion
having arisen from the distinction, between a bacterium and a
bacillus, being made to depend upon length. Observers differed
as to whether a rod of a certain length ought to be considered a
31
SYSTEMATIC.
482
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CLASSIFICATION AND DESCRIPTION OF SPECIES. 483
bacterium or a bacillus. To meet this difficulty a rough-and-
ready rule was suggested—viz., that a rod less than twice its
breadth in length should be considered as a bacterium, and other-
wise a bacillus. But this purely arbitrary division was inadequate,
from the fact that a rod at one stage of its growth or under certain
conditions might, as far as length went, truly be a bacterium, and
under other circumstances be of such a length as to entitle its being
considered a bacillus (Fig. 197). We should avoid such confusion
if we followed Zopf, and acknowledged as a difference between a
bacterium and a bacillus the presence or absence of that form of
spore-formation now distinguished as endogenous spore-formation,
We might then conveniently retain this generic term, to include
that group of rod-forms in which this spore-formation is as yet
t
Fie. 197.—F R1EpLANDER’s PnEuMococcus, x 1500 (Zopf).
unknown; moreover, we should, by so doing, with one or two
exceptions, collect together those short rod-forms which appear to
link the simple cocci to the spore-bearing rods or bacilli.
The grouping together of the different species according to the
character of the colonies in nutrient gelatine is also of questionable
advisability. These characters can hardly be considered to be of
sufficient importance, or indeed in many cases to be sufficiently
constant, to serve by themselves for this purpose. In many cases
a slight variation in the composition of the nutrient medium may
considerably affect the appearances of the colonies. At the same
time, the appearances are very characteristic of certain species of
bacteria, and in some cases the characters of the colonies, together
with the characters of the growth in test-tubes, assist us in
distinguishing species which are morphologically similar, as in the
case of the comma-bacilli of Finkler and of Koch.
484 SYSTEMATIC.
The classification of Zopf will lead the investigator to work upon
the same lines, and by tracing the life-history of individual forms
in pure-cultivations either to extend the work of establishing
protean species or to restrict the doctrine of pleomorphism to a few
forms. For though the author prefers the classification proposed by
Zopf, he is not prepared to accept his views entirely—for instance,
to regard the bacterium of rabbit septicaemia as a micrococcus.
Any arrangement at present can only be considered provisional,
and therefore the most practical classification must be considered
the best. In fact, much more investigation is required before we
can arrive at a permanent and thoroughly scientific classification
of the known bacteria. Many bacteria have been described by
different observers as different species which are really identical.
Many micro-organisms have been described and named as new
species with very insufficient investigation. The determination of
species rests upon the accumulated evidence afforded by a thorough
knowledge of their life-history. The morphological appearances
under different conditions must be carefully studied, the presence
or absence of movement and of spore-formation, and when present
the exact character; the appearances of colonies and of test-tube
cultivations in different media and under different circumstances ;
the liquefaction and other changes in nutrient media; the nature
of the chemical products, if any, and the effect on the living animal
of the bacterium itself and of its products, in varying doses, must
all be taken into account. We must also ascertain whether the
bacterium is an aerobe requiring the presence of oxygen, or an
-enaerobe growing only in the absence of it, or a facultative anuerobe
growing equally well with or without it; and lastly, we must know
whether the bacterium is a parasite requiring a living host, or a
saprophyte existing on dead animal or vegetable matter, or a
facultative parasite capable both of growing in the living animal
and of leading a saprophytic existence. Several writers have
classified the bacteria which have been described hitherto by taking
some of these characters into account, and so preparing a list which
is convenient for the purpose of bacteriological diagnosis. A system
of this kind is of value in leading investigators to supply information
which is wanting in order to verify and amplify the information
upon which the classification is based, and to identify species which
have been described under different names.
CLASSIFICATION OF
SPECIES
FOR BACTERIOLOGICAL DIAGNOSIS
(4) GELATINE NOT
(a) Chromogenic.
Micrococcus violaceus
Micrococcus carneus .
Micrococcus cerasinus siccus
Micrococcus cinnabareus
Micrococcus aurantiacus
Micrococcus versicolor
Micrococcus luteus
Micrococcus citreus
Micrococcus ochroleucus
Micrococcus cereus flavus
Micrococcus agilis citreus .
Micrococcus flavus tardigradus .
Staphylococcus viridis flavescens
COCCI.
LIQUEFIED.
COLOUR.
Violet
Flesh-red .
Cherry-red
Cinnabar-red
Orange
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Greenish- yellow
(b) Non-Chromogenic.
Micrococcus candicans
Micrococcus candidus
Micrococcus concentricus .
Micrococcus fervidosus
Micrococcus cereus albus -
Micrococcus aquatilis
Micrococcus aquatilis invisibilis
Micrococcus cumulatus tenuis .
Micrococcus rosettaceus
Micrococcus viticulosus
Micrococcus plumosus
Micrococcus amylivorus
Micrococcus acidi lactici
Micrococcus uree ¢ é
Micrococcus gingive pyogenes
Micrococcus salivarius septicus
Micrococcus of Manfredi .
Micrococcus in pleuropneumonia
Micrococcus in trachoma .
‘Heematococcus bovis .
Diplococcus albicans tardissimus
HABITAT,
Water.
Water.
Water.
Air; water.
Water.
Water.
Water.
Water.
Water.
Water.
Air.
Air; water.
Lymph.
Air; water.
Water.
Water.
Water.
Pus; water.
Water.
Water.
Nasal mucus.
Water.
Air; water.
Water.
Pear-blight.
Milk.
Air; urine.
Pus.
Saliva.
Sputum.
Lymph.
Conjunctiva.
Urine.
Vaginal secretion
486 CLASSIFICATION OF SPECIES
Diplococcus coryze
Pseudo-diplococcus pncuineate
Micrococcus tetragenus
Micrococcus tetragenus stcirilia wenihtieall
Pediococcus cerevisize
Pediococcus acidi lactici
Streptococcus pyogenes
Streptococcus brevis .
Streptococcus septicus . . ’
Streptococcus vermiformis
Streptococcus of mastitis in cows
Streptococcus in strangles
Streptococcus acidi lactici
(ns) GELATINE LIQUEFIED.
(a) Chromogenic.
COLOUR.
Micrococcus agilis . 3 Pink.
Micrococcus roseus . 5 Pink.
Staphylococcus pyogenes aureus Orange
Micrococcus in pemphigus Orange
Staphylococcus salivarius pyogenes Orange
Micrococcus fuscus. Brown
Micrococcus flavus destdens Yellowish-brown
Micrococcus cremoides Yellowish-white
Micrococcus botryogenus Yellowish.
Micrococcus Finlayensis Pale- ce
Staphylococcus pyogenes citreus Yellow
Micrococcus citreus liquefaciens Yellow
Micrococcus flavus liquefaciens . Yellow
- Micrococcus tetragenus versatilis Yellow
Diplococcus flavus liquefaciens tardus Yellow
Diplococcus subfiavus Yellow
Diplococcus citreus songlomenstus Yellow
Diplococcus luteus Yellow
Diplococcus roseus_. Pink.
Diplococcus fluorescens perides Green
(b) Non-Chromogenic.
Micrococcus albus liquefaciens .
Micrococcus aerogenes
Micrococcus radiatus
Micrococcus feetidus .
Micrococcus in Biskra- iuttinis or Pendijch sore
Micrococcus in influenza
Micrococcus Freudertreichi
Micrococcus in yellow fever
Micrococcus lactis viscosus
Micrococcus acidi lactici liquefaciens
Micrococcus uree liquefaciens .
Micrococcus in gangrenous mastitis in sheep .
Staphylococcus pyogenes albus
Staphylococcus pyosepticus
Diplococcus albicans amplus
HABITAT.
Nasal mucus.
Meningitis.
Sputum.
Stomach.
Beer ; air.
Malt ; hay-dust.
Pus.
Saliva.
Soil.
Water.
Pus.
Pus.
Milk.
Water.
Sputum.
Pus.
Bulle.
Saliva.
Water.
Air ; water.
Water.
Equine tumours.
Yellow fever.
Pus.
Eczema.
Air ; water.
Blood.
Eczema.
Vaginal mucus.
Pus ; air.
Water.
Air.
Posterior nares.
Nasal mucus.
Intestine.
Air ; water.
Nasal mucus.
Pus.
Blood.
Milk.
Blood.
Cream.
Cheesy butter.
Urine.
Milk.
Pus.
Pus.
Vaginal secretion.
FOR BACTERIOLOGICAL DIAGNOSIS.
Pediococcus albus
Streptococcus liquefaciens
Streptococcus septicus liquefaciens
Streptococcus albus .
Streptococcus of Mannaberg
Streptococcus coli gracilis
(c) NO GROWTH IN GELATINE.
Micrococcus pneumoniz croupose
Micrococcus endocarditidis rugatus .
Nitromonas of Winogradsky
Micrococcus in pemphigus
Micrococcus in influenza .
Micrococcus gonorrhoese ‘
Diplococcus intercellularis meuiagiaaie
Micrococcus tetragenus subflavus
Streptococcus giganteus urethre
Streptococcus of Bonome .
487
HABITAT,
Water.
Blood.
Blood.
Water.
Urine.
Feeces.
Saliva.
Heart.
Soil.
Bulle
Sputum.
Pus.
Meningitis.
Nasal mucus.
Urethra.
Meningitis.
(D) GROWTH IN GELATINE UNDETERMINED.
Ascococcus Bilrothii .
Leuconostoc mesenteroides
Streptococcus of progressive tissue necrosis in mice
Micrococcus pyogenes tenuis
Micrococcus of pyeemia in rabbits
Micrococcus of progressive abscess isemalien in mice
Micrococcus of Forbes
Micrococcus in syphilis
Streptococcus Havaniensis
Streptococcus perniciosus psittacorum
Streptococcus bombycis
PACKET COCCI.
(s) GELATINE NOT LIQUEFIED.
(a) Non-Chromogenic.
Sarcina pulmonum
Sarcina ventriculi
(8) GELATINE LIQUEFIED.
(a) Chromogenic.
COLOUR,
Sarcina mobilis . Red
Sarcina rosea Red .
Sarcina flavea Yellow
Sarcina lutea Yellow
Sarcina aurantiaca Orange-yellow .
Meat infusion.
Beet juice; mo-
lasses.
Putrid blood.
Pus.
Meat infusion.
Putrid blood.
Cabbage — cater-
pillars.
Blood
Liver.
Blood of parrot.
Diseased silk-
worms.
Phthisical sputum.
Stomach.
Ascitic fluid.
Air.
Beer.
Air.
Air; water.
488 CLASSIFICATION OF SPECIES
(b) Non-Chromogenic.
HABITAT,
Sarcina alba. i ‘ é é ‘ F Air; water.
Sarcina candida : . : . Air of breweries.
RODS.
(I.) AEROBES OR FACULTATIVE ANAEROBES
(A) GELATINE NOT LIQUEFIED.
(a) Chromogenic.
(4) SPORE-FORMATION PRESENT.
(a) Motile.
COLOUR. ‘
Bacillus cyanogenus Greyish-blue . Blue milk.
- Bacillus erythrosporus ‘ Greenish-yellow . Water.
Bacillus in infantile diarrhea (Lesagey Green Intestine.
(b) Non-Motile.
Bacillus brunneus P Brown . . Water.
(8) SPORE-FORMATION UNKNOWN.
(a) Motitle.
Bacillus rubefaciens Pale-pink Water.
Bacillus rubescens Pale-pink i Sewers.
Bacillus fuscus limbatus . ‘ Brown . : Rotten eggs.
Bacillus beroliniensis Indicus . Indigo-blue Water.
Bacillus cyanogenus Jordaniensis Bluish . Sewers.
Bacillus aurantiacus . Orange . ‘ Water.
Bacillus fluorescens aureus Orange . Water.
Bacillus fluorescens longus . Greyish-yellow Water.
~ Bacillus fluorescens tenuis . . Greenish-yellow Water.
Bacillus constrictus Pale-yellow . Water.
Bacillus subflavus Pale-yellow Water.
Bacillus aureus. i Golden-yellow . Water; eczema.
Bacillus flavescens . Yellow . Swamp-water.
Bacillus heminecrobiophilus Yellowish Caseous glands.
Bacillus canalis parvus Yellowish Sewer-water.
Bacillus fluorescens putidus Greenish . Water.
‘Bacillus dentalis viridans Opalescent-green Carious dentine.
“Bacillus virescens A Green. Sputum.
(6) Non-Motile.
Bacillus latericeus Brick-red Water.
Bacillus spiniferus . Greyish-yellow Eczema.
Bacillus in purpura lesinomhgien Greyish-yellow Blood.
Bacillus uteus . Orange-yellow . Water.
Bacillus in cholera in ducks Yellowish : Blood.
Bacillus flavo-coriaceus Sulphur-yellow Water.
Bacillus striatus flavis . Sulphur-yellow Nasal mucus.
Bacillus fuscus . Deep-yellow . Water.
Bacillus fluorescens non- liquetacions . Greenish-yellow Water.
FOR BACTERIOLOGICAL DIAGNOSIS.
(b) Non-Chromogenic,
(4) SPORE-FORMATION PRESENT.
(a) Motile.
Bacillus putrificus coli
Bacillus septicus vesica
Bacillus in cancer
(6) Non-Motile.
Bacillus acidi lactici (Hueppe) .
Bacillus coprogenes feetidus
Bacillus subtilis similans .
Bacillus epidermidis .
Bacillus of Colomiatti
(8) SPORE-FORMATION UNKNOWN.
(a) Motile.
Bacillus cedematis aerobicus
Bacillus of Fulles (I.)
Bacillus stolonatus
Bacillus venenosus brevis .
Bacillus venenosus é
Bacillus gracilis anaerobiescens
Bacillus invisibilis
Bacillus albus ‘
Bacillus venenosus invisibilis
Bacillus aquatilis sulcatus
Bacillus argenteo-phosphorescens
Bacillus gliscrogenus .
Bacillus cystiformis .
Bacillus of Guillebeau
Bacterium Zopfii
Bacillus ventriculi
Bacillus coli communis
Bacillus cavicida
Bacillus of Utpadel .
Bacillus aerogenes
Helicobacterium aerogenes
Bacterium aerogenes .
Bacillus meningitidis purulentz
Bacillus pyogenes foetidus
Proteus Zenkeri
Bacillus enteritidis
Bacillus hyacinthi septicus
Proteus lethalis . :
Bacillus endocarditidis griseus .
Bacillus of Roth (I.) .
Bacillus Schafferi
Bacillus of swine-plague
Bacillus typhi abdominalis
Bacillus cavicida Havaniensis .
Bacillus cuniculicida Havaniensis
489
HABITAT.
Human feces.
Cystitis.
Stomach.
Sour milk.
Swine measles.
Human feces.
Skin.
Conjunctiva.
Earth.
Earth.
Water.
Water.
Water.
Water.
Water.
Water.
Water
Vater.
Sea-water.
Urine. -
Urine. -
Milk.
Intestine ; air.
Stomach of dogs.
Intestine.
Intestine.
Intestine.
Intestine.
Intestine.
Intestine.
Pus.
Pus.
Putrid substances.
Poisonous meat.
Rotten hyacinths.
Septicemia.
Endocarditis.
Old rags.
Cheese ; potato.
Lymphaticglands.
Spleen and glands.
Intestine.
Intestine
490 CLASSIFICATION OF SPECIES
(0) Non-Motile.
Bacillus of Okada
Bacillus pyogenes soli
Bacillus of Fulles (II.)
Bacillus candicans
Bacillus septicus agrigenus
Bacillus scissus .
Bacillus ubiquitus
Bacillus multipediculus
Bacillus albus anaerobiescens
Bacillus Zurnianus
Bacillus canalis capsulatus
Bacillus of Roth (II.)
Bacillus tenuis sputigenus
Bacillus crassus sputigenus
Bacillus coprogenes parvus
Bacillus of Fiocca
Bacillus striatus albus
Bacillus capsulatus mucosus
Bacillus pseudo- ee nt
Bacterium ure
Bacillus nodosus parvus
Bacillus oxytocus perniciosus .
Bacillus lactis pituitosi
Bacillus limbatus acidi lactici
Bacillus ovatus minutissimus
Bacillus of Belfanti and Pascarola
Bacillus of Tommasoli
Bacillus capsulatus .
Bacillus of purpura hosmorrhagiéa (Babes)
Bacillus of purpura hemorrhagica (Kolb)
Bacillus septiceemiz hemorrhagice .
Proteus capsulatus septicus
Bacillus acidiformans
Bacillus coli similis .
Bacillus hepaticus fortuitus
Bacillus filiformis Havaniensis
Bacillus of Martinez.
Bacillus diphtherize columbraram
Bacillus diphtherie .
Bacillus of Schimmelbusch
Bacillus capsulatus Smithii
Bacillus erysipelatis suis .
Bacillus in rhinoscleroma .
Bacillus of Friedlinder
Bacillus pneumosepticus .
Bacillus endocarditidis capsulatus
Bacillus septicus keratomalaciz
Bacillus of intestinal diphtheria in rabbits
Bacillus of acne contagiosa of horses
Bacillus pseudo-tuberculosis
Bacterium tholeeideum
Bacillus gallinarum .
HABITAT.
Dust.
Earth.
Soil,
Soil.
Manured soil.
Soil.
Air; water.
Air; water.
Water.
Water.
Sewer-water.
Old rags.
Sputum.
Sputum.
Feeces.
Saliva.
Nasal mucus.
Nasal secretion.
Healthy throat.
Urine.
Healthy urethra.
Milk.
Milk.
Milk,
Eczema.
Pus.
Hair with sycosis.
Blood.
Blood.
Blood.
Blood.
Blood.
Liver.
Liver.
Liver.
Liver.
Liver.
Diphtheritic de-*
posit.
Diphtheritie
throat.
Cancrum oris.
Intestine.
Blood.
Rhinoscleroma.
Sputum.
Septic pneumonia,
Endocarditis.
Internal organs.
Intestine.
Pus.
Internal organs.
Intestine.
Blood.
FOR BACTERIOLOGICAL DIAGNOSIS.
Bacillus argenteo-phosphorescens ..
Bacillus smaragdino-phosphorescens
Bacillus phosphorescens gelidus
Proteus hominis capsulatus
Bacillus of grouse disease.
Bacillus lactis aerogenes .
(4) GELATINE LIQUEFIED.
(a) Chromogenic.
(A) SPORE-FORMATION PRESENT.
(a) Motile.
Bacillus violaceus
Bacillus in disease of bees (Canestrini)
Bacillus in ‘‘red-cod ”
Bacillus mesentericus ruber
Bacillus fluorescens _ liquefaciens
minutissimus .
COLOUR.
Deep-violet
Pink.
Red .
Reddish-yellow
Greenish-yellow
(8) SPORE-FORMATION UNKNOWN.
(a) Motite.
Bacillus ianthinus
Bacillus violaceus TLiereaklis
Bacillus lividus .
Bacillus carnicolor
Bacillus rubidus .
Bacillus Indicus .
Bacillus rosaceus mietatiniden
Bacillus ochraceus
Bacillus citreus cadaveris .
Bacillus buccalis minutus .
Bacillus arborescens .
Bacillus fulvus
Bacillus plicatilis
Bacillus pyocyaneus .
Bacillus fluorescens Liqnefacians
Bacillus cyanofuscus .
Bacillus fluorescens nivalis
Bacillus chromo-aromaticus
Bacillus viscosus.
Bacillus pyocyaneus .
Bluish-violet
Deep-violet
Violet-black
Dark flesh-colour
Brownish-red .
Sealingwax-red
Magenta-red
Yellow
Yellow
Yellow
Yellow
Yellow
Vellowis.
Yellowish-green
Greenish-yellow.
Greenish-brown
Bluish-green
Green or brown
Green
Green
(b) Non-Motile.
Bacillus cceruleus
Bacillus glaucus .
Bacillus membranaceus amethystinus
Bacillus lactis erythrogenes
Bacillus mycoides roseus
Blue.
Grey.
Violet
Red .
Red .
491
HABITAT,
Fish.
Fish.
Cuttlefish.
Blood.
Blood.
Intestine.
Water.
Larvee of bees.
Salted codfish.
Potatoes.
Skin.
Water.
Water.
Water.
Water.
Water.
Intestine.
Water.
Water.
Blood.
Saliva.
Water.
Water.
Water.
Pus.
Water.
Cheese ; glue.
Water.
Intestine.
Water.
Pus.
Water.
Water.
Water.
Milk.
Soil.
492 CLASSIFICATION OF SPECIES
Bacillus prodigiosus .
Bacillus hydrophilus fuscus
Bacillus cuticularis
Bacillus helvolus
Bacillus tremelloides
Ascobacillus citreus
COLooR,
Blood-red .
Yellow
Yellow
Yellow
Yellow
Yellow
Bacillus igenkeo. phos ghovcseons
liquefaciens
Bacterium termo (Vignal) .
Bacillus smaragdinus foeetidus
Yellowish
Yellowish
Green
(b) Non-Chromogenic.
(4) SPORE-FORMATION PRESENT.
Bacillus infiatus
Urobacillus Freudenreichi_
Bacillus mycoides -
Bacillus ramosus
Bacillus gracilis .
Bacillus circulans
Urobacillus Duclauxi
Urobacillus Maddoxi
Bacillus limosus .
Bacillus butyricus
Bacillus Hessii .
Bacillus lactis albus .
Bacillus liodermos
Urobacillus Pasteuri .
Bacillus mesentericus fuscus
Bacillus mesentericus vulgatus .
Bacillus of potato rot
Bacillus maidis .
Bacillus megatherium
Bacillus tumescens
Bacillus subtilis .
Bacillus subtilis similis
Bacillus vacuolosis
- Bacillus of Scheurlen
Bacillus alvei
Bacillus aerophilus
Bacillus implexus
Bacillus filiformis
Bacillus vermicularis
Bacillus incanus
Bacillus inunctus
Bacillus granulosus
Bacillus carotarum
(a) Motile.
(b) Non-Motile.
HABITAT.
Air.
Frog’s lymph.
Water.
Water.
Water.
Skin.
Sea-water.
Saliva.
Ozzena.
Air.
Air ; dust 5 sewers.
Soil; water.
Soil; water.
Water.
Water.
Water.
Water.
Sea-dredgings.
Milk.
Milk.
Milk.
Milk.
Urine.
Potato; dust;
water.
Potato; water, etc..
Rotting potatoes.
Maize infusion.
Boiled cabbage.
Beet-root.
Dust ; water ; soil.
Liver.
Liver.
Cancerous tissues.
Larve of bees.
Air.
Water.
Water.
Water.
Swamp-water.
Swamp-water.
Sea-dredgings.
Boiled carrot.
FOR BACTERIOLOGICAL DIAGNOSIS,
Bacillus brassicze
Bacillus in gangrene .
Bacillus of Letzerich .
Bacillus anthracis
(8) SPORE-FORMATION UNKNOWN.
(a) Motile.
Bacillus pestifer
Bacillus diffusus
Bacillus gasoformans
Bacillus liquidus
Bacillus guttatus
Bacillus liquefaciens .
Bacillus radiatus aquatilis
Bacillus nubilis . 4
Bacillus albus putidus
Bacillus hyalinus
Bacillus vermiculosus
Bacillus delicatulus
Bacillus punctatus
Bacillus reticulans
Bacillus figurans CVn ind
Urobacillus Schutzenbergi
Bacillus devorans
Bacillus venenosus liqudliciens:
Bacillus aquatilis
Proteus sulphureus
Bacillus stoloniferus .
Bacillus phosphorescens frdiews
Bacillus phosphorescens indigenus
Bacillus cyaneo-phosphorescens
Bacillus litoralis
Bacillus halophilus
Bacillus superficialis .
Bacillus cloace .
Proteus microsepticus
Proteus vulgaris.
Proteus mirabilis
Proteus septicus
Bacillus foetidus ozenz ,
Bacillus septicus ulceris gangrenosi .
Bacillus albus cadaveris
Bacillus of Guillebeau
Bacillus Havaniensis liquefaciens
Bacillus carabiformis P -
Bacillus of Schou
Bacillus leporis lethalis
Bacillus liquefaciens communis
(0) Non-Motile.
Bacillus buccalis fortuitus .
493
HABITAT.
Cabbage infusion.
Senile gangrene.
Urine.
Blood.
Air.
Soil.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Water.
Swamp-water.
Sea-water.
Sea-water.
Sea-water.
Sea-dredgings.
Sea-dredgings.
Sewage.
Sewage.
Uterine dis-
charges,
Putrid substances.
Putrid substances.
Septicemia.
Nasal mucus.
Blood and organs.
Blood.
Milk.
Skin.
Stomach.
Rabbit.
Liver.
Liver.
Saliva.
494 CLASSIFICATION OF SPECIES
HABITAT.
Bacillus ulna (Vignal) Saliva.
Leptothrix buccalis (Vignal) Mouth.
Bacillus varicosus conjunctive . Conjunctiva.
Bacillus gingive pyogenes Alveolar abscess.
Bacillus pulpe pyogenes . Gangrenous tooth-
pulp.
Bacillus graveolens ° Skin of feet.
Pneumo-bacillus liquefaciens bovis . Lung.
(c) NO GROWTH IN GELATINE.
(A) SPORE-FORMATION PRESENT.
(a) Motite.
Bacillus ulna. . é White of egg.
Bacillus in putrid bronchitis Sputum.
Bacillus mallei . Glandered tissue.
(b) Non-Motile.
Bacillus in erythema nodosum . Blood.
Bacillus tuberculosis . Tubercular tissue-
(8) SPORE-FORMATION ABSENT.
Bacillus sanguinis typhi Blood.
Bacillus septicus acuminatus Septic infection.
Bacillus necrophorus . Condyloma.
(c) SPORE-FORMATION NOT STATED.
,
Bacillus allantoides Air.
Bacillus nitrificans Soil.
Bacillus in measles Blood.
Bacillus in ophthalmia Conjunctiva.
(D) GROWTH IN GELATINE NOT STATED.
Bacillus senilis .
Bacillus leptosporus .
Bacillus allii
Bacillus indigogenus .
Blood.
Air.
Putrefying onions.
. Infusion of indigo.
Il. ANAEROBES.
(4) GELATINE LIQUEFIED.
Bacillus rubellus
Bacillus butyricus (Botkin)
Clostridium foetidum
(4) SPORE-FORMATION PRESENT,
(a) Motile.
Dust.
Milk;water; dust
Earth.
FOR BACTERIOLOGICAL DIAGNOSIS,
‘Bacillus radiatus
Bacillus liquefaciens magnus
Bacillus spinosus
Bacillus thalassophilus
Bacillus of symptomatic anthrax
Bacillus edematis maligni
Bacillus tetani. .
(6) Non-Motile.
Bacillus liquefaciens parvus
Bacillus anaerobicus liquefaciens
(8B) GELATINE NOT LIQUEFIED.
(4) SPORE-FORMATION PRESEN.
(a) Motile.
Bacillus amylozyma .
Bacillus solidus .
Bacillus polypiformis
(b) Non-Motile.
Bacillus muscoides
(8) SPORE-FORMATION ABSENT.
(b) Non- Motile.
Bacillus aerogenes capsulatus
495
HABITAT.
Earth.
Earth.
Earth.
Sea-dredgings,
Tissues (quarter-ill).
Lymph.
Wounds ; earth.
Earth.
Yellow fever.
Water.
Earth.
Earth.
Earth.
Blood.
(c) GROWTH IN GELATINE NOT STATED.
(4) SPORE-FORMATION PRESENT.
(a) Motile.
Bacillus butyricus . : .
(3) SPORE-FORMATION UNKNOWN.
Bacillus cadaveris
CURVED RODS.
(a) GELATINE NOT LIQUEFIED.
(a) Chromogenic.
(a) Motile.
: COLOUR.
Spirillum rubrum ‘ ‘ . Deep-red
(b) Non-Chromogenic.
Spirillum suis
Vegetable infu-
sions, etc.
Liver.
Putrid mouse.
Intestine of swine.
496 CLASSIFICATION OF SPECIES
Spirillum concentricum
Spirillum saprophiles
(b) Non-Motile.
(a) Chromogenic.
COLOUR.
Spirillum flavescens Yellowish-green
Spirillum flavum Ochre-yellow
Spirillum aureum Orange-yellow .
(b) Non-Chromogenic.
Spirillum lingue
Spirillum nasale
GELATINE LIQUEFIED.
(a) Non-Chromogenic.
(a) Motile.
Spirillum cholere Asiatice
Spirillum of Finkler and Prior.
Spirillum Metchnikovi
Spirillum of Miller
Spirillum of Sanarelli
Spirillum tyrogenum
Spirillum marinum
HABITAT.
Putrefying blood.
Hay-infusion;
sewage.
Sewers.
Sewers.
Sewers.
Deposit on tongue.
Nasal mucus.
Intestine.
Intestine.
Intestine of fowls.
Carious teeth.
Water.
Old cheese.
Sea-dredgings.
NO GROWTH IN GELATINE, OR UNDETERMINED.
(@) Motitle.
Spirillum Obermeieri
Spirillum anserum
Spirillum undula
Spirillum sputigenum
Spirillum serpens
Spirillum tenue .
Spirillum volutans
Spirillum plicatile
(b) Non-Motile.
Spirillum dentium
Spirillum sanguineum
Blood.
Blood of geese.
Putrid infusions.
Gums.
Stagnant water.
Putrid infusions.
Swamp-water.
Swamp-water.
Gums.
Brackish water.
BRANCHING FILAMENTS.
Streptothrix actinomycotica.
Streptothrix alba.
Streptothrix liquefaciens.
Streptothrix musculorum suis.
Streptothrix Hofmanni.
Streptothrix farcinica.
Streptothrix asteroides.
Streptothrix carnea.
FOR BACTERIOLOGICAL DIAGNOSIS. 497
Streptothrix aurantiaca.
Streptothrix chromogenes.
Streptothrix odorifera.
Streptothrix violacea.
Streptothrix Forsteri.
Streptothrix madure.
NOT CLASSIFIED.
Bacillus indigonaceus.
Bacillus proteus fluorescens.
Beggiatoa alba.
Beggiatoa mirabilis.
Beggiatoa roseo-persicina.
Cladothrix dichotoma.
Cladothrix invulnerabilis.
Crenothrix Kuhniana.
Diplococcus citreus liquefaciens.
Leptothrix buccalis.
Leptothrix gigantea.
Micrococcus aquatilis invisibilis.
Micrococcus crepusculum.
Micrococcus feetidus.
Micrococcus Havaniensis.
Micrococcus of septicemia in rabbits.
Monas Okenii.
Monas vinosa.
Monas Warmingii.
Myconostoc gregarium.
Rhabdomonas rosea.
Sarcina hyalina.
Sarcina intestinalis,
Sarcina litoralis.
Sarcina Reitenbachii.
Sarcina urine.
Spherotilus natans.
Spirillum attenuatum.
Spirillum leucomelaneum.
Spirillum rosaceum.
Spirillum Rosenbergii.
Spirillum rufum.
Spiromonas Cohnii.
Spiromonas volubilis.
Streptococcus cadaveris.
Streptococcus flavus desidens.
Vibrio rugula.
32
DESCRIPTION OF
SPECIES ARRANGED FOR
REFERENCE IN ALPHABETICAL ORDER.
Ascococcus Billrothii—Small
globular cocci, united into charac-
teristic colonies.
They form on the surface of
nourishing fluids a cream-like skin,
divisible into an enormous number
of globular or oval families. Each
family is surrounded by a thick
capsule of cartilaginous consistency.
In a solution containing acid tar-
trate of ammonia the fungi generate
butyric acid, and change the origin-
ally acid fluid into an alkaline one.
Fig. 198.—Ascococcts BILLROTHIT
(Cohn).
They were first observed on putrid
broth, and later on ordinary nourish-
ing solutions; they also readily de-
velop upon damp slices of boiled
roots, carrots, beetroots, etc.
Ascobacillus citreus (Unna,
Tommasoli).— Rods sometimes
curved, 1°3 u in length, ‘3 » in width,
singly, in pairs, and masses. The
colonies develop slowly, and are
yellowish in colour.
The cocci inoculated in the depth
of gelatine form small colonies in
the track of the needle, anda slimy
pale-yellow growth on the surface ;
liquefaction sets in slowly.
On agar the growth is gelatinous,
and orange in colour, and rapidly
extends over the surface.
On potato the growth is abundant,
and pale yellow.
They were isolated from the skin
in eczema seborrhceicum.
Bacillus acidiformans (Stern-
berg).—Short rods, 1:5 to 3 pw in
length, 1°2 « in width, and filaments.
5 to 10 p.
Colonies circular ; iridescent by
reflected light.
Inoculated in the depth of gela-
tine they grow freely in the track of
the needle, and form a hemispherical
mass on the surface. They produce
gas bubbles.
On agar the growth is milk-white,
and the jelly becomes strongly acid.
On potato the growth is abundant.
In broth with 5 per cent. glycerine
they produce opacity and a copious
viscid deposit, and the surface is
covered with gas bubbles,
Injected into the peritoneal
cavity of rabbits and guinea-pigs,
they produce death in twenty-four
hours,
They were isolated from the liver
in a fatal case of yellow fever.
Bacillus acidi lactici (Hueppe).
—Rods 1 to 2°8 » long, and ‘3 to
‘4 » wide, and thread forms. Spore-
formation present. In_ gelatine
498
DESCRIPTION OF SPECIES.
cultures the breadth of the rods is
diminished. They grow best be-
tween 35° and 42° C., and cease
under 10° C. Over 45°5° C. they
no longer produce acidity,
Whitish colonies appear on the
second day.
In gelatine a delicate growth
appears along the whole track of
the needle, with spherical forms
here and there.
In milk they produce lactic acid
and the casein is precipitated.
Bacillus aerogenes (Miller).—
Small rods varying in length.
Colonies white or yellowish-white ;
concentric.
In the depth of gelatine they
produce a yellowish filament, and
on the surface a grey patch with
dentated periphery ; later the fila-
ment is brown.
On potato the growth is yellowish
and dry.
They were isolated from the in-
testine in health.
Bacillus aerogenes capsulatus
(Welch).—Rods straight or slightly
curved, 3 to 6 yw; threads and
chains ; capsulated.
Colonies on agar greyish-white,
with hairy processes.
They peptonise gelatine and pro-
duce gas. Broth becomes turbid,
and there is an abundant sediment.
Milk is coagulated. Cultures have
a faint smell of glue.
Injected into rabbits they pro-
duce gas in the blood and internal
organs.
They were isolated from a patient
after death, with blood-vessels full
of gas.
Bacillus aerophilus (Liborius).
—Rods and filaments.
Colonies punctiform ;
yellow.
Inoculated in the depth of gela-
tine the bacilli produce a funnel of
liquefied jelly, with flocculi in the
lower part.
On potato they form a smooth
yellowish layer.
They were isolated from con-
taminated cultures.
Bacillus albus (Hisenberg).—
Rods and chains.
greyish-
499
Colonies circular, white.
In gelatine the bacilli grow in the
track of the needle, and form a
white hemispherical mass on the
free surface.
On agar the growth is pure white,
and on potato yellowish- white.
They occur in water.
Bacillus albus anaerobiescens
(Vaughan).—Short rods.
Colonies circular,
brown.
Inoculated in the depth of gela-
tine they grow in the track of the
needle, and on the free surface.
On agar the growth is pure white,
and on potato yellowish-white.
They occur in water.
Bacillus albus cadaveris
(Straussmann and Stricker).—Rods
2-5 win length, ‘75 » in width, and
filaments.
Colonies yellowish ; circular, and
later radiated.
Inoculated in the depth of gela- -
tine they produce a funnel of lique-
fied gelatine with a thick deposit.
On agar there is an abundant
white growth.
Ou potato the growth is white or
yellowish-white, and colours the
potato in the vicinity bluish-brown.
The cultures have a putrefactive
odour.
Mice inoculated subcutaneously
die in six hours, and guinea-pigs in
twenty-four.
They were isolated from putrid
human blood.
Bacillus albus putidus (De
Bary).—Rods and filaments.
Colonies circular and brownish.
Inoculated in the depth of gela-
tine they . produce rapid lique-
faction. >
On agar and potato the growth
is slimy. Cultures develop a strong
putrefactive odour.
They occur in water.
Bacillus allantoides (L. Klein).
—Rods 2 to 2°5 p in length, ‘5 » in
width, and in chains. The rods
develop cocci-forms united by a
gelatinous substance into zoogleea
masses. They were isolated from
a contaminated culture.
Bacillus allii (Griffiths).—Rods
yellowish-
500 DESCRIPTION
5 to 7 » in length, 2°5 » in width,
singly, in pairs, and zoogloea.
On agar they produce a bright
green film, and cultures are said to
emit traces of sulphuretted hydro-
gen.
, They were isolated from putrid
onions.
Bacillus alvei (p. 470).
Bacillus amylozyma (Perdrix).
—Rods 2 to 3 » in length and ‘5 p
in width, in pairs, and in chains.
They are anaerobic.
Colonies white, and producing gas
bubbles.
On potato in an atmosphere of
hydrogen the bacilli partly liquefy
it, and there is abundant formation
of gas.
They ferment sugar and starch.
Bacillus anaerobicus lique-
faciens (Sternberg).—Slender rods,
about ‘6 » in diam., in pairs, and
in filaments.
Colonies granular and white ; sur-
rounded by liquefied gelatine.
They grow along the track of
the needle when inoculated in the
depth of agar.
They were isolated from the in-
testine in a fatal case of yellow
fever.
Bacillus anthracis (p. 192).
Bacillus aquatilis (Frankland).
—Rods 2°5 » in length, and filaments
17 » or longer. They resemble
Bacillus arborescens.
Colonies after liquefaction of the
gelatine have a yellowish-brown
nucleus from which proceed twisted
strands of filaments.
Inoculated in the depth of gela-
tine the growth in the track of the
needle is at first almost invisible,
later liquefaction occurs.
On agar the growth is shining
and yellowish.
Broth becomes turbid, and a
sediment forms at the bottom of
the tube.
On potato there is a slightly
yellowish streak.
They occur in water.
Bacillus aquatilis fluorescens
(Lustig).—Short thin rods with
rounded ends. Non-motile.
Colonies fern-like and iridescent.
4
OF SPECIES.
Compare Eisenberg’s Bacillus fluo-
rescens non-liquefaciens.
Bacillus aquatilis graveolens
(Tataroff).—Slender rods 1:3 » in
length. They rapidly liquefy gela-
tine and produce an odour like that
of perspiration from the feet.
They occur in water.
Bacillus aquatilis sulcatus
(Weichselbaum), No. I.—Rods mor-
phologically, and in cultures resem-
bling Bacillus typhosus.
Colonies in gelatine exhibit lin
and furrows.
The growth on the surface of
gelatine is said to be greater than in
cultures of Bacillus typhosus grown
for comparison.
They occur in water,
No. IJ.—Rods also resembling in
morphology and cultivation the
Bacillus typhosus.
Colonies are said to be thicker than
those of No. I., and not dentated.
The growth on potatoes is yel-
lowish-brown, and emits a faint
odour of urine.
They occur in water.
No. III.—Very short rods.
Colonies show lines and furrows,
and are yellowish,
On the surface of gelatine the
growth develops as a thin whitish
film. ;
On agar the growth is white and
abundant.
On potato the growth is yellow.
They were isolated from water.
No. IV.—Rods and filaments.
Colonies circular and bluish.
On the surface of gelatine the
growth is greyish-white, and on
agar there is a similar appearance.
They do not grow on potato,
They occur in water.
No. V.—Rods rather thicker than
those of Bacillus typhosus.
Colonies similar to those of No. I.
The growth on the surface of
gelatine is yellow.
On agar the growth is viscid and
yellow, and on potato the growth is
faintly yellow and the surrounding
medium stained bluish-grey.
They occur in water.
Bacillus arborescens (Frank-
land)—Rods 2°5 p in length, and
es
DESCRIPTION OF SFECIES.
‘5 p» in width, singly, in pairs, and
in short chains, and filaments.
Colonies throw out delicate
branches with a highly characteristic
appearance ; the gelatine slowly
liquefies, the nucleus of the colony
becomes yellow, and the periphery
iridescent.
Inoculated in the depth of gela-
tine the bacilli form a cloudiness in
the track of the needle, and an
iridescent layer on the surface with
central depression of the gelatine
and commencing liquefaction.
Later the liquefaction produces
a funnel, and there is a yellow
deposit.
On the surface of agar the layer
is a dirty orange colour.
On potato the growth is orange
red with irregular protuberances,
and limited in growth.
They occur in water.
Bacillus argenteo-phosphores-
cens (Katz), No. I.—Rods slightly
curved with pointed ends, 2°5 » in
length, width one-third of their
length ; singly, in pairs, and long
wavy filaments.
Colonies circular ; at first trans-
parent droplets, later yellowish in
colour.
On the surface of gelatine they
form a greenish-yellow film.
In broth they produce turbidity,
and later a skin on the surface, and
on sterilised fish a pale-yellow sticky
growth.
Cultures are photogenic.
They were isolated from the sea
at Sydney.
No. II.—Rods with rounded ends
-27 » in length, 67 » in width, and
filaments.
Colonies on gelatine are circular
with sharp contours and greyish-
yellow in colour; later they are
irregular and granular.
Inoculated in gelatine the bacilli
form a greyish-white filament in
the track of the needle, and a shining
patch on the surface,
On the surface of obliquely solidi-
fied gelatine they form a bluish-
grey film.
In broth they produce only
turbidity.
501
Cultures are photogenic.
They were isolated from phospho-
rescent fish.
No. IfI.—Rods not so thick as
those of No. IT., singly, in pairs, and
filaments. They are motile.
Colonies are white, scaly, and
wrinkled.
On the surface of gelatine the
growth spreads over the medium.
On agar the growth is scanty.
In broth they produce turbidity
and a skin floating on the surface.
Cultures are photogenic after a
few days’ growth.
They were isolated from a piece
of cuttle-fish.
Bacillus argenteo-phosphores-
cens liquefaciens.—Rods straight
or slightly bent, 2 » in length, and
in width one-third of their length ;
filaments.
Colonies circular, pale brown or
pale yellow and, after liquefaction,
with radiating processes extending
into the surrounding gelatine.
Inoculated in the depth of gela-
tine there is a growth in the track
of the needle, and near the surface
a cup-sbaped area of liquefaction.
In broth they produce turbidity,
and form a skin on the surface.
On sterilised fish they form a
yellow layer.
They are photogenic but not
markedly so.
Bacillus aurantiacus (Frank-
land).—Rods short and thick, singly,
in pairs, and in filaments.
Colonies are prominent and pale
orange in colour.
Inoculated in the depth of gela-
tine there is a slight growth in the
track of the needle and an orange
patch on the free surface.
On agar and potato the growth
is also orange.
They occur in water.
Bacillus aureus (Adametz).—
Slender rods straight or slightly
bent, 1:5 to 4 » in length, and ‘5 »
in width; in pairs, filaments, and
masses. They are motile.
Colonies circular or oval and
yellow in colour.
Inoculated in the depth of gela-
tine the growth is very limited in
502
the track of the needle, while
small chrome-yellow hemispherical
masses develop on the free surface.
On potato they form a chrome-
yellow growth.
They occur in water and on the
skin.
Bacillus berolinensis Indicus
(Clissen).—Slender rods, singly, in
pairs, and short chains ; capsulated.
Colonies at first whitish acquire
in a few days an indigo-blue colour.
On the surface of gelatine they
form a blue layer, which slowly
spreads.
On the surface of agar the indigo-
blue colour is very marked.
On potato they grow abundantly,
and develop the same colour. The
pigment is insoluble in alcohol,
chloroform or water, soluble in
strong acids, and decolorised by
ammonia.
They were isolated from river
water at Berlin.
Bacillus brassice (Pommer).—
Rods 1:9 to 5-4 » in length, 91 to
1:2 » in width, and filaments. They
form spores.
Colonies have the appearance of
a fine mycelium.
Inoculated in the depth of gela-
tine the growth in the track of the
needle sends off fine filaments, and
liquefaction quickly follows.
In the depth of agar white
colonies form in the track of the
needle, and on the surface the
growth is first cloudy and later
yellowish.
They were isolated from infusion
of cabbage.
Bacillus brevis (Mori).—Rods
‘25 pw long and ‘8, broad. Non-
motile.
Colonies pale-yellow ; non-lique-
fying.
Inoculated in the depth of gela-
tine small dots appear along the
needle track and a pale yellowish
growth on the surface.
On agar at 35° C. a yellowish
and on blood serum a greyish
growth appears in two or three
‘days. They do not grow on potato.
In broth they form a white cloudy
deposit.
DESCRIPTION OF SPECIES.
Mice inoculated subcutaneously
die in from sixteen to thirty hours.
They are also pathogenic in guinea-
pigs and rabbits.
They were found in drain water.
Bacillus brunneus (Adametz
and Wichman).—Rods small and
slender. Spore formation present.
Colonies at first white, later
brownish.
Inoculated in the depth of gela-
tine the growth occurs along the
track of the needle and also on
the surface, developing a brownish
colour in the surrounding gelatine.
They occur in water.
Bacillus buccalis fortuitus
(Vignal)—Rods 1:4 to 3 mw in
length, singly, and in pairs.
Colonies circular and liquefying.
Inoculated in the depth of gela-
tine liquefaction occurs slowly, and
white flocculi occur in the liquid,
and later subside to the bottom. -
In broth they produce turbidity
and a skin on the surface.
They were isolated from saliva.
Bacillus buccalis maximus
(Miller).—Rods 2 to 10 » in length,
1 to 13 win width, and filaments
30 to 150 » in length. j
They occur in the mouth.
Bacillus buccalis minutus (Vig-
nal).—Rods 5 to 1 p» in length,
and slightly less in width.
Colonies circular and faintly
yellow.
Inoculated in the depth of gela- -
tine they form a yellowish-white
growth in the track of the needle,
and a patch of the same colour on
the surface; liquefaction com-
mences slowly, and extends down-
wards until the gelatine is com-
pletely liquefied and a yellow mass
collects at the bottom of the tube.
In broth there is a similar deposit,
and an iridescent pellicle on the
‘surface.
On potato they form a yellow film.
They were isolated from saliva.
Bacillus butyricus, (Praz-
mowski. Bacillus amylobacter, Van
Tieghem ; Bacillus of butyric acid
JSermentation).—Rods 3 to 10 p» long,
under 1 » wide, often indistinguish-
able from Bacillus subtilis. They
DESCRIPTION
grow out into long, apparently un-
jointed threads. They are mostly
actively motile, but also occur in
zoogloeea. The rods and threads
are sometimes slightly bent, like
vibrios. They are anaerobic. The
shorter rods asa rule swell in the
middle, becoming ellipsoidal, lemon,
A
"0 b¢) 7) "Ca
Fic. 199.—CLostRIDIUM BUTYRICUM.
A. Active stage. (a, b) Bent rods (vibrio-
form) and threads. (c) Short
rods. (d) Long rods. .
B. Spore-formation. C. Spore-germina-
tion. (Prazmowski.)
or spindle-shaped ; the long rods,
and sometimes the short ones, swell
at one end ; in either case ellipsoidal
‘Spores are developed (Fig. 199).
Cultivated in nutrient gelatine,
the medium is liquefied, and a scum
formed on the surface. They grow
h Ny
OF SPECIES. 503
best between 35° and 40° C. The
spores are widely distributed in
nature, and grow readily on fleshy
roots, old cheese, etc. They convert
the lactic acid in milk into butyric
acid, and produce the ripening of.
cheese.
They occur also in solutions of
starch, dextrine, and sugar, and are
the active agents in the fermenta-
tionof sauerkrautand sour gherkins.
Bacillus butyricus (Botkin).—
Rods and filaments, spore-formation
present. They are anaerobic.
Colonies consist of felt-like
masses.
Inoculated in the depth of gela-
tine with 1:5 per cent. of grape-
sugar, the growth commences in
the lower part of the needle track
with abundant formation of gas
bubbles and liquefaction of the
jelly.
In milk there is abundant gas
formation, which will break the
flasks if closed.
They were isolated from milk,
earth, and water.
Bacillus butyricus
(Hueppe).—Rods slightly bent,
21 » in length, 38 » in width,
and filaments.
Colonies yellowish; rapidly
liquefying.
Inoculated in the depth of
gelatine liquefaction occurs
along the track of the needle, and
later a wrinkled skin floats on
the surface.
On agar the growth is yel-
lowish.
On potato the growth is
wrinkled and faintly yellow.
They coagulate milk, precipita-
ting and then dissolving the casein.
They occur in milk.
Bacillus cadaveris (Sternberg),
—Rods 1° to 4 p» in length, and
1:2 » in width, singly, in pairs, and
in filaments.
They are anaerobic. Colonies in
glycerine-agar are irregular, granu-
lar, and white.
They produce an acid reaction
in cultures. :
Subcutaneous injection in guinea-
pigs may produce extensive
504 DESCRIPTION
oedema and death in twenty-four
hours.
They were obtained from the
liver in fatal cases of yellow fever.
Bacillus ceruleus (Smith).—
Rods 2 to 2°5 pin length, and ‘5 p
in width, singly and in chains.
Colonies blue.
Inoculated in the depth of gela-
tine they form a colourless growth
in the track of the needle, and a
cup-shaped cavity in its upper part,
with bluish contents.
On agar they form a blue layer
and a deep-blue growth on potato.
They occur in water.
Bacillus canalis capsulatus
(Mori).—Rods ‘9 to 1°6 » 1m width,
capsulated.
Colonies milk-white.
The growth in the depth of gela-
tine is similar to Friedlander’s
pneumococcus.
On agar the growth is viscid, and
on potato it is yellowish.
In broth a skin forms on the
surface.
They are pathogenic in mice.
They occur in sewage.
Bacillus canalis parvus (Mori).
—Rods 2 to 5 » in length, ‘8 tol p
in width.
Colonies very minute, pale yellow.
On the surface of gelatine they
very slowly form a yellowish film.
On agar the growth is dry and
yellow.
They are pathogenic in mice and
guinea-pigs.
They occur in sewage.
' Bacillus candicans (Frankland).
—Very short rods and filaments.
Colonies pure white.
Inoculated in the depth of gela-
tine they form isolated colonies in
the track of the needle, and a white
button on the free surface.
On agar they form a greyish
layer, and flourish on potato.
They occur in soil.
Bacillus capsulatus (Mori).—
Oval forms and rods, sometimes
encapsuled. Non-motile. Colonies
white.
_ Inoculated in the depth of gela-
tine and agar a nail-shaped growth
occurs.
OF SPECIES.
In broth they form a white
| turbidity, and a white pellicle
develops on the surface and on the
sides of the vessels.
On potato at 37°C. an abundant
moist, yellowish, stringy growth is
formed, with production of gas
bubbles. They are pathogenic in
mice and in rabbits if injected into-
the pleural cavity.
They occur in drain water.
Bacillus capsulatus (Pfeiffer)..
—Rods singly, in pairs, or in chains
and in filaments. They have a well-
marked capsule. Colonies white.
Inoculated in gelatine they grow
in the track of the needle, and
form a white button on the free
surface.
On agar and on potato the growth
is also white and very viscid, so-
that it can be drawn out into long
threads.
They produce a fatal result in
mice in two or three days, when
inoculated subcutaneously. A.
minute quantity of a broth culture
injected into the peritoneal cavity
of guinea-pigs will prove fatal in.
thirty-six hours. The bacilli are
found in the blood which is made
viscid.
They were isolated from a guinea-
pig found dead.
Bacillus capsulatus mucosus.
(Fasching).—Rods 3 to 4 » in
length, ‘75 to 1 » in width ; capsu--
lated. Colonies white.
Cultures in gelatine resemble
Friedlinder’s pneumococcus.
They form gas.
They produce a fatal result in
mice in thirty-six hours.
They were isolated from nasal
mucus in cases of influenza.
Bacillus capsulatus suis.
(Smith).—Rods from 1:2 to 1°8 p
in length and ‘8 to ‘9 » in width.
There are three varieties of this
bacillus having a closeresemblance to
the pneumococcus of Friedlander.
They were isolated from the in-
testines of swine.
Bacillus carabiformis (Kac-
zynsky).—Rods short and slender.
Colonies develop characteristic
processes.
DESCRIPTION
Inoculated in the depth of gela-
tine, the bacilli produce liquefaction
and colour the liquid greenish-
yellow.
On agar they form a yellowish-
white layer.
They were isolated from the
stomach of a dog.
Bacillus carnicolor (Tils).—
Rods 2 » long, and °5 yp broad.
Singly actively motile. Spore-
formation not observed.
Colonies are in the form of cup-
shaped depressions.
Inoculated in the depth of gela-
tine they grow rapidly along the
whole track of the needle, forming
a funnel-shaped area of liquefaction
at the bottom of which there is a
pale pink deposit.
On potato they form slowly a
dark flesh-coloured growth.
They occur in water.
Bacillus carotarum (A. Koch).
—Rods ‘97 to 1:05 p» in lengthand
filaments.
Colonies white and circular.
Inoculated in the depth of gela-
tine the bacilli grow slightly in the
track of the needle and abundantly
on the surface.
On agar they form a white, and
on potato a faintly brown layer.
They occur on boiled carrot and
beet.
Bacillus cavicida (Brieger).—
Rods morphologically and in culti-
vations similar to Bacillus coli com-
munis, |
Cultures: are said to be patho-
genic in guinea-pigs.
They were isolated from feces.
Bacillus cavicida Havaniensis
(Sternberg).—Rods 2 to 3 mw in
length, and ‘7 » in width, singly
and in pairs.
Coionies are of a pale-straw
colour. :
Inoculated in the depth of gela-
tine the bacilli form small trans-
lucent pearl-like spherical colonies,
and on the free surface the growth
is limited.
On potato the growth is at first
thin and dirty yellow, and later
gamboge yellow.
Guinea-pigs inoculated subcuta-
OF SPECIES. 505
neously die
hours.
They were isolated from the
intestinal contents in a fatal case
of yellow fever, by inoculation of
guinea-pigs.
Bacillus chromo-aromaticus.—
Rods which liquefy gelatine and
form a yellowish-white scum on
the surface.
On potato the growth is irides
cent and brownish.
In broth a scum forms on the
surface and the broth is coloured
greenish-blue. Cultures have an
aromatic odour.
They are said to produce pneu-
monia and pleurisy in rabbits.
They were isolated from a pig
with post-mortem appearances of
swine-fever.
Bacillus circulans (Jordan).—
Rods 2 to 5 w'in length and 1 p» in
width, singly and in short chains.
Colonies are brownish.
Inoculated in the depth of gela-
tine they liquefy the medium in
the track of the needle, forming
a conical cavity at its upper part.
On agar they form a translucent.
film.
Milk is slowly coagulated.
In broth they produce turbidity
and a slimy deposit.
They occur in water.
Bacillus citreus cadaveris
(Strassmann).—Rods °9 » in length,
‘6 w» in width, singly and in chains.
Colonies pale yellow.
Inoculated in the depth of gela-
tine the bacilli form minute colonies
along the track of the needle, and
at its upper part liquefy the gela-
tine and produce a yellow deposit.
They were found in the blood
after death.
Bacillus cloace (Jordan).—
Short rods ‘8 to 1:9 » in length,
‘7 to 1 » in width, singly and in
pairs.
Colonies circular, yellowish.
Inoculated in the depth of gela-
tine liquefaction occurs in the track
of the needle, an iridescent scum
forms on the surface, and there is
an abundant deposit. ;
On agar the growth is milk-
in ten or twelve
506
white, and on potato yellowish-
white.
In broth they produce turbidity
and ascum on the surface. They
reduce nitrates,
They occur in sewage.
Bacillus coli communis
(Escherich).—See p. 344,
Bacillus coli similis (Stern-
berg).—Rods 1 to 3 p in length, ‘4 to
‘5 p in width ; singly and inpairs.
Colonies circular and pale brown
in colour.
In the depth of gelatine they
form a scanty growth in the track
of the needle, and on the free sur-
face a translucent film with irregu-
lar margins,
On potato the growth is pale
brown or dirty white.
They were isolated from human
liver after death.
Bacillus constrictus (Zimmer-
mann).—Rods from 1°5 to 6°5 pw in
length, and ‘75 w in width. The
rods are segmented.
Colonies are circular, granular,
and greyish-yellow.
In the depth of gelatine they
form a filament in the track of the
needle and irregular yellow heaps
on the free surface.
On the surface of agar the growth
consists of a yellow shining layer,
and on potato the same colour is
produced.
They occur in water.
Bacillus coprogenes fetidus
(Schottelius).—Rods aboutas large
as Bacillus subtilis, but shorter.
They are non-motile. Spore-for-
mation occurs when the bacilli have
access to the air, but not in the
animal body.
In the depth of gelatine a fila-
ment forms composed of yellow-
ish compact colonies; and on the
surface a fine transparent film;
cultures emit a strong putrefactive
odour.
On potato they form alight grey,
dry layer. : :
Subcutaneous injection of small
doses had no effect on mice and
rabbits, but very large quantities
produced a toxic effect in rabbits.
Swine are not affected.
DESCRIPTION OF SPECIES.
They were found by Schottelius
in the intestine in cases of swine
erysipelas. —
Bacillus coprogenes parvus
(Bienstock).—Very short rods.
On the surface of gelatine they
form a very limited, almost in-
visible, growth in the track of the
needle.
In mice they produce oedema and’
death in thirty-six hours, and in
rabbits a local rash and death in
eight days.
They were isolated from human
evacuations. j
Bacillus crassus aromaticus
(Taratoff).—Rods 3°5 to 5 yw long,
1:5 » in width ; constricted in the
centre.
Colonies appear in the form of
cup-shaped depressions and pro-
duce a fruit-like odour.
In gelatine they grow in the
track of the needle, and later
produce a funnel-shaped area of
liquefaction.
They occur in well water.
Bacillus crassus sputigenus
(Kreibohm).—Short thick rods,
sometimes curved; capsulated.
Colonies greyish-white. Cultures
in gelatine resemble those of Fried-
lander’s pneumococcus. They are
pathogenic in smallanimals. They
were isolated from human sputum.
Bacillus cuniculicida, Bacillus
of Rabbit Septicemia (see p. 228).
Bacillus cuticularis (Tils).—
. Rods from 2 to 3 w in length, 3 to
‘5 » in width, and filaments.
Colonies are yellow and the gela-
tine is liquefied.
In the depth of gelatine they
produce liquefaction, and a skin
forms on the surface.
On potato the growth is slimy
and yellow. ae:
They occur in water. .
Bacillus cuticularis albus
(Taratoff).—Rods 3:2 » long, con-
stricted in the middle. Actively
motile. :
Colonies are opalescent and
bluish-white.
Inoculated in the depth of gela-
tine they form a white rosette-
shaped growth on the surface and
DESCRIPTION OF SPECIES.
a shining white growth
along the needle track
which sends off long
rounded processes.
On agar, glycerine agar
and blood serum they
produce a luxuriant white
‘shining growth.
Broth becomes turbid
with a whitish deposit and
pellicle.
Qa potato there is a thick
moist brown growth.
They are found in water.
Bacillus cyaneo-fuscus
(Beyerinck).—Rods °2 to -6
p in length, and half their
length in width. Motile.
In the depth of gelatine
they produce colonies in
the track of the needle,
which later develop a black
pigment.
In broth with 3 per cent.
of peptone they produce a
blue colour, changing to
brown and finally black.
They were isolated from
cheese, size and glue.
Bacillus cyaneo-phosphores-
eens (Katz).—Rods 2°6 » in length,
1 » in width, singly, in pairs, and
filaments. Colonies circular and
brownish or greyish-yellow.
Inoculated in the depth of gela-
tine they form a grey-white fila-
ment in the track of the needle and
liquefaction follows at the upper
part ; later a skin forms on the sur-
face and a yellow deposit occurs at
the bottom of the liquefied jelly
which has a reddish-brown tinge.
In broth a similar skin floats
on the surface and the broth is
turbid.
On sterilised fish the growth is
viscid and yellow. ;
Cultures are phosphorescent,
especially in media containing
excess of common salt.
They were isolated from sea-
water at Sydney, and are possibly
identical with Bacillus phosphores-
ens of Fischer.
Bacillus cyanogenus (Hueppe),
—Bacterium syncyanum. Bacillus
of Blue Milk.—Motile rods, 2°5 to
short rods.
¥F, G. Spore-forming rods. H.
forms (NEELSEN).
ia) Cc A
AiG wy
gs dria !
f off 4 M7
i
Fic. 200.—BacILLUS CYANOGENUS, x 650. A. Ac-
tive rods. B. Rods in zooglea, C. Chain of
D. Chain of cocci. E. Cocci stage.
Involution-
3°5 w in length, and ‘4 » wide (Fig.
200). The rods after division may
remain linked together, and form
chains. Non-motile rods occur
enveloped in a gelatinous capsule,
and involution forms.
Colonies appear after two days
as small greyish-white points which
gradually assume a moist appear-
ance. The gelatine becomes steel-
grey, throwing the white colonies
into strong relief.
In the depth of gelatine a whitish
growth appears in the track of the
needle near the upper part, and on
the free surface, producing also a
dark steel-blue discoloration of
the jelly which spreads down-
wards.
On agar a greyish growth ap-
pears, and the agar is coloured dark
brown. ,
On potato a yellowish moist
growth develops, the potato around
it is stained grey-blue. Milk be-
comes slightly alkaline and of a
slate-grey colour, which on the
addition of acid changes to an
intense blue. Milk in which the
508 DESCRIPTION
lactic acid bacillus is growing be-
comes sky-blue from the first.
They are non-pathogenic.
They are present in blue milk.
Bacillus cyanogenus (Jordan).
—Rods 1:3 » in length, °8 p in
width. Slightly motile.
Colonies granular and irregular.
They colour the surrounding gela-
tine brown.
Inoculated in the depth of gela-
tine the bacilli produce a scanty
growth in the track of the needle,
and a film on the surface with
coloration of the gelatine beneath
it.
On agar they form a white layer,
and the jelly is coloured brown.
On potato the growth is brown.
They were isolated from sewage.
Bacillus cystiformis (Clado).—
Short slender rods. Motile.
Colonies circular, yellowish, gra-
nular.
Inoculated in the depth of gela-
tine there is a scanty growth in the
track of the needle, and a white
patch on the free surface.
On agar the growth is yellowish-
white.
They were isolated from urine.
Bacillus delicatulus (Jordan).
—Rods 2 » long and 1 p» broad,
often in pairs or short chains.
Actively motile. Spore-formation
not observed.
Colonies at first whitish with
a radiating edge. Later they
liquefy the gelatine and the centres
become dark.
Inoculated in the depth of gela-
tine they rapidly liquefy it and
form a whitish pellicle and a brown
deposit.
On agar a greyish crinkled
growth appears, which gradually
becomes white and shining.
On potato there is a grey flat
growth.
Milk is coagulated and becomes
strongly acid.
Broth is made turbid and a white
serum and precipitate formed.
They occur in sewage.
Bacillus dentalis viridans
(Miller).—Rods slightly bent, singly
and in pairs.
OF SPECIES.
Colonies circular, yellowish and
concentric.
Inoculated in the depth of gela-
tine they grow in the track of the
needle and on the free surface, and
the jelly is coloured green:
On agar the growth is colourless.
or slightly grey.
Intraperitoneal injections in mice
and guinea-pigs produce a fatal re-
sult. Subcutaneous injectidns cause
suppuration.
They were isolated from caries.
of the teeth.
Bacillus dentriticus (Bordoni
Uffreduzzi and Lustig).—Rods -85-
to 2°8 w long, and ‘5 to °85 y» broad,
singly and in zooglea. Motile.
Spore-formation not observed.
Colonies have an arborescent.
appearance.
Inoculated in the depth of gela-
tine they form a circular raised
growth at the point of puncture,.
and white colonies along the needle
track. The jelly is gradually
liquefied.
On agar and blood serum there
is a scanty growth on the surface
and an abundant growth in the
track of the needle. Blood serum
is liquefied after some time.
Broth is rendered turbid; a
white firm pellicle forms.
On potato there is a thick moist
‘white growth, which later becomes
yellow.
Found in water.
Bacillus devorans (Zimmer-
mann).—Rods -99 » to 1:2 p in
length, 74 » in width; singly, in
pairs and in chains.
Colonies are circular, granular,
and grey, with periphery formed of
radiating processes.
In the depth of gelatine they
produce a whitish filament and an
excavation at the upper part, which
may or may not contain liquefied
jelly.
On agar a greyish film is found.
They do not grow on potato.
They occur in water.
Bacillus diffusus (Frankland).
—Rods 1:7 yp in length, °5 p» in
width ; singly, in pairs, and fila-
ments.
DESCRIPTION OF SPECIES.
Colonies circular, bluish-green,
with a granular nucleus and delicate
irregular periphery.
In the depth of gelatine there is
scarcely any growth in the track of
the needle, but a shining greenish-
yellow film on the surface, and
liquefaction below it.
On agar the growth is faintly
yellow.
In broth they produce turbidity
and a yellowish deposit.
On potato the growth is yellow-
ish.
They occur in earth.
Bacillus diphtherie (p. 332).
Bacillus diphtherie colum-
barum (p. 336).
Bacillus dysodes(Zopf).—Cocci,
Jong and short rods, and spores.
They were observed in bread,
making it greasy and unfit for food,
and generating a penetrating odour
resembling a mixture of pepper-
mint and turpentine. <A great loss
may result to bakers if the fungus
is introduced with the yeast.
Bacillus endocarditidis cap-
sulatus (Weichselbaum),—Cocci
resembling Friedlinder’s pneumo-
coccl.
Colonies faintly yellow,
dentated contours.
In the depth of gelatine the
growth produces a filament in the
track of the needle, and a patch like
stearin on the free surface.
Large doses injected subcutane-
ously, or into the peritoneal cavity
prove fatal to rabbits.
They were isolated from infarcts
in a fatal case of endocarditis.
Bacillus endocarditidis griseus.
—Rods motile.
Colonies granular and brown or
yellowish-brown.
In the depth of gelatine there is
a filamentous growth in the track
of the needle, and a circular whit-
ish patch on the surface. On agar
and potato the growth is greyish-
brown.
Cultures cause a fatal result in
mice and guinea-pigs.
They were isolated from a case of
endocarditis.
Bacillus enteritidis (p. 372).
with
509
| _ Bacillus epidermidis (Bordoni
| Uffreduzzi).—Rods 2°8 to 3 pw in
| length, and 3» in breadth. Spore-
| formation occurs at 25° C.
| They grow very sparingly on
gelatine.
On agar there is a surface growth.
On potato at 15° to 20°C. the
growth appears first in the form
of drops, which gradually extend
and coalesce and form a thin layer
| Fic. 201.—Pure Curtivation or Ba-
| cCILLus FIGURANS ON THE SURFACE
| ov NUTRIENT AGAR-AGAR.
over the surface, On blood serum
| they form a thin film.
Inoculation in rabbits and guinea-
pigs and on the human skin pro-
duced no result.
They were isolated from flakes
of cuticle from between the toes.
Bacillus erysipelatis suis (p.
356).
Bacillus erythrosporus
(Eidam). — Slender rods and
510 DESCRIPTION OF SPECIES.
filaments, Motile. Spore-forma- On potato the growth is brown-
tion present. They occur in water.
Fic. 202.—PHorocraPH or Part or AN IMPRESSION PREPARATION OF
Bactntus Ficurans ON NuTRIENT GELATINE, x 50.
Colonies circular, with brown nu- Bacillus figurans (Crookshank).
cleus and yellowish-green periphery. | —Rods, with rounded ends, varying
In the depth of gelatine they | inlength. Spore-formation present.
é
f
?
Fic. 203.—Part or THE SAME SPECIMEN SHOWN IN Fic. 202 x 200.
grow in the track of the needle In plate-cultivations they cause
and on the free surface, colouring | a cloudy growth, spreading from
the jelly green by transmitted light, | various points; if a cover-glass
yellow by reflected light. impression is made, this is found
DESCRIPTION OF SPECIES.
to consist of the regularly-arranged
parallel rods. The chains of rods
become twisted at intervals into
curious convolutions, from which
offshoots are continued in various
directions. These long shoots or
processes are again twisted at
intervals into varying shapes and
patterns (Figs. 202, 203). Culti-
vated in nutrient gelatine, the bacilli
form on the surface visible windings,
from which fine filaments grow
down into the gelatine. They
spread out also in almost parallel
lines transversely from the needle
track. The gelatine is not liquefied.
On an oblique surface of nutrient
agar-agar the filaments spread down-
wards into the substance of the
jelly, and outwards from the cen-
tral streak on the surface, forming
a feather-like cultivation (Fig. 201).
They were obtained from the
air, and later were identified by
the author with Bacterium Zopfii..
Bacillus figurans (Vaughan).—
Rods and threads.
Colonies composed of curved and
nterlacing lines.
In gelatine they grow in the track
of the needle, and very slowly
liquefy it.
On agar they form a thin white
layer.
They were found in water.
Bacillus filiformis (Tils).—Rods
4 » in length and 1 mw in width,
singly and in chains. Spore-forma-
tion present.
Colonies are granular, with yel-
lowish nucleus.
Inoculated in the depth of gela-
tine there is no growth in the
track of the needle, but a whitish
growth on the surface, liquefying
the jelly slowly. |
In broth they form a skin on
the surface.
On agar the growth is white.
On potato the growth is dry and
after a time brownish. They
coagulate milk.
They occur in water. :
Bacillus filiformis Havaniensis
(Sternberg).—Long slender rods,
3 p» in diam., and filaments.
Colonies circular, irregular ; deep
ol}
colonies are brownish ; superficial
colonies thin and translucent.
Inoculated in the depth of gela-
tine the growth is scanty in the
track of the needle.
In the depth of agar an opaque
branching growth occurs in the
track of the needle, and a scanty
milk-white growth on the surface.
In broth they cause opalescence.
They were isolated from the
liver in fatal cases of yellow fever.
Bacillus flavescens (Pohl).—
Rods 2:1 to 2°2 w in length, ‘8 p.
in width. Slightly motile.
Colonies yellow and granular.
Inoculated in the depth of gela-
tine the bacilli produce a filament
in the track of the needle, and a
growth spreads over the free sur-
face.
On the surface of agar the growth
is composed of isolated yellow
colonies.
On potato they grow rapidly,
forming a shiny yellow layer.
They occur in marsh water.
Bacillus flavocoriaceus (Ada-:
metz).—Minute rods occurring in
zoogloea.
Colonies circular, sulphur-yellow.
Under a low power they show a
brownish-yellow nucleus and yellow
periphery.
Inoculated in the depth of gela-
tine the growth is granular in the
track of the needle and on the free
surface.
They occur in water.
Bacillus fluorescens aureus
(Zimmermann).—Short rods, 1:9 p
in length, ‘74 p in width.
Singly, in pairs and in masses.
Motile ; flagellated.
Coloniescircular, granular, yellow.
Inoculated in the depth of gela-
tine they form a filament in the
track of the needle and a yellow
patch on the surface.
On agar the growth is golden-
yellow, and the same on potato.
They occur in water.
‘Bacillus fluorescens lique-
faciens (Fliigge).—Short rods with
rounded ends.
Colonies on plates develop an
iridescence around them.
512 DESCRIPTION
In the depth of gelatine a white
filament forms in the track of the
needle, with liquefaction in , the
upper part, and an iridescent sheen
is produced in the jelly.
On potato they develop a brown-
ish layer.
They occur in water and in
putrid infusions.
Bacillus fluorescens lique-
faciens minutissimus (Unna and
Tommasoli).—Rods 1°5 to 2 » in
length, ‘3 » in width.
Colonies circular, with brownish
nucleus and yellowish marginal
zone.
In the depth of gelatine lique-
faction forms in the track of the
needle. The liquid is turbid and
yellowish, and a white sediment
forms at the bottom and a fluores-
cent scum on the surface.
On agar and potato the growth is
brownish.
They were isolated from eczema-
tous skin.
Bacillus fluorescens longus
(Zimmermann).—Rods varying in
length, some curved, 1°45 to 1°65 p
in length, °83 » in width, and wavy
filaments 14 » in length. Motile.
Colonies circular, well-defined,
yellowish, with broad twisted
markings.
On the surface of gelatine they
produce a layer with a bluish-green
fluorescent colour.
On agar a thin film forms, and
the jelly is coloured greenish-
yellow.
On potato the growth is slimy
and yellowish.
They occur in water.
Bacillus fluorescens nivalis
(Schmolck)—Rods and chains.
Motile.
Colonies circular, surrounded by
liquid fluorescent jelly.
Inoculated in the depth of gela-
tine they produce liquefaction in
the track of the needle, and the
jelly is coloured green.
On agar the growth is white and
the jelly green.
On potato the growth is brownish.
They are probably identical with
Bacillus fluorescens liquefaciens.
\
OF SPECIES,
They were isolated from snow.
Bacillus fluorescens non-lique-
faciens (Eisenberg)—Short deli-
cate rods. Non-motile. ;
Colonies have the appearance
of fern-leaves, and are opales-
cent.
Inoculated in the depth of gela-
tine the growth is very slight in
the track of the needle, and on the
surface filmy and fluorescent.
On agar they form a greenish
layer.
They produce a brownish layer
on potato, and bluish-grey dis-
coloration.
They occur in water.
Bacillus fluorescens putidus
(Fliigge).—Short rods with rounded
ends. Motile ; spore-formation not
known.
They form small dark colonies
with a greenish sheen, which have
a penetrating odour.
Inoculated in the depth of gela-
tine they produce a_pale-grey
growth, and after three days colour
the medium with a greenish tinge
spreading down from above.
On potato they rapidly develop
a brownish layer.
They occur on decomposing sub-
stances, producing a greenish
coloration.
According to Lehmann and Neu-
mann the Bacillus fluorescens albus
and Bacillus fluorescens longus of
Zimmermann and Bacillus fluores-
cens non-liquefaciens are merely
varieties of Bacillus fluorescens
putidus ; and further, cultures of
this bacillus on agar and on potato
and in milk and in broth cannot
be distinguished from those of
Bacillus fluorescens liquefaciens.
Bacillus fluorescens tenuis
(Zimmermann).—Rods 1 to 1°85 uh
in length, ‘8 » in width. Singly,
in masses, and filaments. Motile.
Colonies irregular.
Inoculated in the depth of gela-
tine a delicate filament forms in
the track of the needle, and on
the surface a greyish-white growth
spreads and colours the gelatine
yellow.
On agar the growth is shining
DESCRIPTION
-and greenish, and on potato yellow-
ish.
They occur in water.
Bacillus fetidus (Bacterium
fetidum, Thin).—Cocci, short rods,
long rods, and leptothrix. The
cocci, 1:25 to 1:4 y in diam., occur
singly or in pairs. Spore-formation
present in the rods.
They were isolated from the
exudation in a case of profuse
sweating of the feet,.and the odour
was noticeable in the cultivation
(vide Bacillus saprogenes).
Bacillus fetidus ozene (Ha-
jek).—Short rods, singly, in pairs,
and in short chains. Motile.
Colonies irregular and liquefying.
In the depth of gelatine lique-
faction occurs along the track of
the needle.
On agar the growth is moist and
shiny, and on potato yellowish-
brown.
Cultures emit a disagreeable
odour.
They produce a fatal result in
mice and local inflammation in
rabbits.
They were isolated from cases
of ozeena.
Bacillus fulvus (Zimmermann).
—Rods 88 to 1:3 yw in ITength,
‘77 w in width. Singly, in pairs
and in chains.
Colonies vary in form ; granular,
yellowish-grey.
In the depth of gelatine there is
a scanty faintly yellow growth in
the track of the needle, and a
hemispherical yellow mass on the
free surface..
On agar and potato the growth
is yellow and shining.
They occur in water.
Bacillus fuscus (Zimmermann).
—Rods ‘63 » in width, varying in
length ; sometimes bent and irregu-
lar in form.
Colonies irregular, granular, and
greyish-yellow.
In the depth of gelatine a
hemispherical mass appears on the
free surface, which later spreads
and forms a yellow wrinkled layer.
On agar and potato the layer is
similarly coloured,
OF SPECIES. 513
They occur in water.
Bacillus fuscus limbatus
(Scheibenzuber).—Rods and fila-
ments.
Colonies with brown nucleus and
light periphery.
Inoculated in the depth of gela-
tine the growth in the track of the
needle is branching and the jeliy
coloured brown.
On agar and potato the growth
is dark brown.
They occur in rotten eggs.
Bacillus gallinarum (Bacillus
of Fowl Enteritis) (see p. 230).
Bacillus gasoformans (Eisen-
berg).—Rods. ,
The colonies are granular, and
liquefy the gelatine.
The bacilli inoculated in‘ the
depth of gelatine rapidly produce
liquefaction in the track of the
needle, and formation of gas
bubbles.
They occur in water.
Bacillus gingive pyogenes:
vide Bacterium gingiva pyogenes.
Bacillus glaucus (Maschek).—
Rods.
The colonies are well-defined,
greyish in colour, and after a time
liquefy the gelatine.
The bacilli inoculated in the
depth of gelatine produce a rapid
growth in the track of the needle
and on the surface, followed by
liquefaction and a sediment at the
bottom of the liquid.
On potato and agar they form a
greyish layer.
They occur in water.
Bacillus gliscrogenus (Malerba).
—Rods ‘57 to 1:14 win length, 41 »
in width.
Colonies spherical, granular.
The bacilli inoculated in the
depth of gelatine give rise to a
growth in the track of the needle
composed of closely packed disc-
shaped colonies.
On agar they produce an opales-
cent film.
On potato they form a viscid
yellowish growth.
They were isolated from viscid
urine.
Bacillus (Zimmer-
33
gracilis
514
mann).—Rods sometimes curved,
2:4 to 3:6» in length, ‘77 » in width,
and filaments.
The colonies are greyish or
yellowish-grey, with concentric
markings.
The bacilli inoculated in the
depth of gelatine produce isolated
colonies in the track of the needle,
and a translucent film on the free
surface ; followed after some time
by slight liquefaction.
On agar they produce a bluish-
white layer.
There is scarcely any growth on
potato,
They occur in water.
Bacillus gracilis anaerobies-
cens (Vaughan).—Rods.
The colonies are brownish.
The bacilli inoculated in the
depth of gelatine produce a copious
growth in the track of the needle,
and gas bubbles, and a film on the
surface.
On agar they produce a thin
layer, and on potato a yellowish-
white mass.
They occur in water.
Bacillus granulosus (Russell).
—Rods singly, in pairs and in
masses, and long filaments. Spore-
formation present.
The colonies have concentric
linear markings.
The bacilli inoculated in the
depth of gelatine cause slow lique-
faction spreading downwards in
the track of the needle.
On the surface of agar they form
more or less isolated whitish or
yellowish colonies.
In broth they produce turbidity,
and on potato a thick shining layer,
which is at first white and later
brownish.
They were isolated from deep-sea
dredgings.
Bacillus graveolens (Bordoni-
Uffreduzzi).—Very short rods, ‘8 »
in length.
The colonies are greyish-white,
and liquefy the gelatine.
The bacilli inoculated in the
depth of gelatine produce rapid
liquefaction in the track of the
needle. They colour the gelatine
DESCRIPTION OF SPECIES.
greenish-yellow, and produce a foul
odour.
On potato the culture is brown-
ish.
They were isolated from skin
from between the toes.
Bacillus guttatus (Zimmer-
mann).—Rods 1 to 1-13 » in length ;
‘93 in width ; singly, in pairs, and
in chains.
Colonies bluish-grey ; granular.
The bacilli inoculated in the
depth of gelatine develop colonies
in the track of the needle, and a
greyish opalescent film on the
surface.
On agar the growth is greyish-
white. On potato it is yellowish
and slimy.
They occur in water.
Bacillus halophilus (Russell).—
Rods 1°5 to 3°5 w in length, ‘7 » in
width ; singly and in pairs; and
toruloid involution forms. Motile.
The colonies liquefy gelatine and
become frothy from abundant for-
mation of gas.
The bacilli inoculated in the
depth of gelatine grow in the track
of the needle, and excavate the
jelly at the upper part.
They were isolated from deep-
sea dredgings.
Bacillus Hansenii (Rasmussen).
—Rods 2°8 to 6 pw long, 6 to 8 p
wide.
Cultivated on sterilised potato,
they form in four days a chrome-
yellow layer with an agreeable
fruit-like smell. Two or three days
later the growth dries, and changes
to an orange-yellow colour ; later
it becomes yellowish or brown, and
at the same time spores are formed
1:7 wlong, 1:1 wide. The colour-
ing matter is insoluble in most
reagents.
The bacilli occur as a yellow or
whitish skin on nourishing solu-
tions, malt infusion, broth, and
wine, which have been kept at
31° to 33° C.
Bacillus Havaniensis lique-
faciens.—Rods ‘8 » in width, 1:2
to 5 p» in length, singly, and in
pairs ; and filaments. Motile.
The colonies are milky, irregular
DESCRIPTION
in outline, and liquefy the gela-
tine.
The bacilli inoculated in the
depth of gelatine cause liquefaction
in the track of the needle.
On agar they form a pale-brown
layer.
They do not grow on potato.
They were isolated from the skin.
Bacillus helvolus (Zimmer-
mann).—Rods 1°5 to 4'5 win length,
‘5 » in width; in pairs, and in
chains.
Colonies are pale yellow.
The bacilli form a yellow growth
on the surface of gelatine, and pro-
duce slow liquefaction.
On agar the growth is yellow.
They occur in water.
Bacillus _heminecrobiophilus
(Arloing).—Rods highly polymor-
phic, and filaments 1 to 20 » in
length. Slightly motile.
On the surface of obliquely
solidified gelatine they form a
yellowish layer.
On potato the growth is yellowish-
white.
They produce cedema when sub-
cutaneously inoculated in the
vicinity of wounds.
They were isolated from a caseous
lymphatic gland in a guinea-pig.
Bacillus hepaticus fortuitus
(Sternberg). — Rods _ resembling
Bacillus coli communis.
The colonies, marked with radia-
ting striz, are dark brown in colour.
The bacilli inoculated in the
depth of gelatine produce a very
slight growth at the upper part of
the track of the needle, and a hemi-
spherical mass on the free surface.
On potato they form a creamy
white growth.
They were isolated from the
liver in a fatal case of yellow
fever.
Bacillus Hessii (Guillebeau).—
Rods 8 to 5 » in length, 1:2 » in
width, cocci-forms and filaments.
The colonies are filamentous,
and liquefy gelatine.
The bacilli inoculated in the
depth of gelatine produce lique-
faction, and the liquid jelly is made
extremely viscous.
OF SPECIES. 515
On potato the growth is brownish.
They coagulate milk.
They were isolated from milk.
Bacillus hyacinthi septicus
(Heinz).—Rods 4 to 6 y in length,
1 w in width.
The colonies are transparent and
bluish-white.
The bacilli inoculated in the
depth of gelatine produce a fila-
ment in the track of the needle
and a layer on the surface.
On potato they produce a slimy,
dirty-yellow layer.
They were isolated from diseased
hyacinths.
Bacillus hyalinus (Jordan).—
Rods 3°6 to 4 in length, 15 p» in
width, and chains. ;
The colonies are surrounded by
radiating filaments.
The bacilli inoculated in the
depth of gelatine produce liquefac-
tion in the track of the needle, a
scum on the surface and a deposit
at the bottom.
On agar they produce a dry, grey
growth.
On potato the growth is greyish-
white and tuberculated.
They coagulate milk.
In broth they produce turbidity
and a pellicle on the surface; and
they are powerful nitrifying
agents.
They occur in water.
Bacillus hydrophilus fuscus
(Sanarelli)—Rods 1 to 3 p» in
length, and filaments 15 to 20 » in
length.
They rapidly produce a funnel-
shaped area of liquefaction when
grown in gelatine, followed by
complete liquefaction and _ the
formation of a white flocculent
deposit.
Inoculated in glycerine agar they
grow rapidly and produce gas
bubbles. On potato they produce
a straw-coloured layer which be-
comes distinctly yellow and later
brown.
They are pathogenic in cold-
blooded animals and in small warm-
blooded animals. Guinea-pigs suc-
cumb in twelve hours; the spleen
is enlarged and the bacilli are found
516 DESCRIPTION
in great numbers in the blood and
internal organs.
They were isolated from the
lymph of diseased frogs.
Bacillus ianthinus (Bacterium
ianthinum Zopf, Bacillus violaceus).
—Slender rods, about four times
their width in length, with rounded
ends. They also form threads, and
are actively motile. Spore-forma-
tion present in the rods.
The colonies occur as circum-
scribed liquefied areas, in the centre
of which is a collection of the
coloured growth.
The bacilli inoculated in the
depth cf gelatine produce a funnel-
shaped liquefaction, and a granular-
looking violet mass subsides to the
bottom.
On agar-agar and potato a
beautiful violet growth rapidly
develops. The colouring matter is
soluble in alcohol.
They were observed oun pieces of
pigs’ bladder floating on the surface
of water rich in bacteria. They
occurred only on the surface of the
bladder exposed to the air, and never
on the part under water. They occa-
sionally occur in common tap water.
Bacillus implexus (Zimmer-
mann). Rods 2:5 » in length,
115 » in width. Non - motile.
Spore-formation present.
Colonies white, granular, develop-
ing in three days into masses of
interlacing white filaments.
Inoculated in the depth of gela-
tine, a growth develops in the track
of the needle and fine filaments
penetrate the gelatine. The jelly is
liquefied, and a pellicle forms on
the surface, and there is a flocculent
deposit.
On agar the growth is white, and
on potato yellowish-white.
They occur in water.
Bacillus in acne contagiosa in
horses (Dieckerhoff and Grawitz).
—Short rods ‘2 » in diam.
Inoculated in the depth of
nutrient gelatine they form a scanty
growth in the track of the needle
and a white patch on the free sur-
face. They thrive best on blood
serum and on agar.
OF SPECIES.
The bacilli inoculated on the
surface of the skin of horses, calves
and other animals are said to pro-
duce acne pustules. Inoculated
subcutaneously in guinea-pigs they
produce a fatal result in twenty-
four hours.
' They were isolated from pus
in cases of acne contigiosa in horses.
Bacillus in cancer (Koubasoff).
—Rods ; spore-formation present.
Inoculated in the depth’ of gela-
tine an irregular filament develops
in the track of the needle and a
transparent growth with central
depression on the surface.
They are said to be pathogenic
in small animals, and to produce
nodules and ulcers of the mucous
membrane of the stomach.
They were isolated from a case
of cancer of the stomach.
Bacillus in cholera in ducks
(Cornil and Toupet), p. 230.
Bacillus in choleraic diarrhea
(Bovet).—Rods 2 to 4 » in length,
1 to 1:5 » in width, singly and in
pairs, and filaments.
In the depth of gelatine a filament
forms in the track of the needle and
a greyish transparent layer on the
surface.
On agar a greyish film is formed.
On potato the growth is yellowish
and abundant.
Intra-peritoneal injections in
guinea-pigs cause peritonitis and
death.
They were isolated from a case
of choleraic diarrhoea.
Bacillus indiphtheritic disease
of calves (Bacillus vitulorum
Loffler).— Rods about five or six
times as long as wide, mostly united
in long threads.
A piece of membrane from a diph-
theritic disease in a calf, placed on
blood serum developed a white layer
composed of the bacteria. Succes-
sive generations were not obtainable.
Mice inoculated directly from the
calf died of a characteristic illness,
and the same long bacteria were
again found in the imoculated
animals accompanying widespread
infiltration, starting from the point
of inoculation. Inoculation of
DESCRIPTION OF SPECIES.
guinea-pigs and rabbits gave doubt- |
ful results. They were found in
the deeper stratum of pseudo-
diphtheritic patches in calves.
Bacillus in disease of bees (p.
471).
Bacillus in erythema nodosum
(Demme).—Rods 2:2 to 2°5 w in
length, ‘5 to ‘7 » in width. They
can be cultivated at 37°C.
Colonies on agar are white with
radiating lines.
The bacilliinoculated in the depth
of agar grow in the track of the
needle, and produce peculiar off-
shoots in the surrounding jelly.
They are said to produce an
eruption resembling erythema
nodosum when inoculated subcu-
taneously in guinea-pigs.
They were obtained from the
eruption and the blood in cases of
erythema nodosum.
Bacillus in fowl enteritis
(Klein), p. 230.
Bacillus in gangrene (Tricomi).
—Rods 3 p» in length, 1 » in width,
singly and in pairs.
Colonies circular, granular, dirty-
yellow.
In the depth of gelatine they
produce a filament composed of
closely aggregated colonies, and at
the upper part conical liquefaction
of the jelly, beneath a cup-shaped
excavation.
On agar and potato the growth
is white.
Injected subcutaneously in rabbits
and guinea-pigs they produce gan-
grene and death in a few days.
They were isolated from a case
of senile gangrene.
Bacillus in grouse disease
(Klein), p. 230.
Bacillus in hog cholera (p.
351).
Bacillus in infantile diarrhea
(Booker).— Rods morphologically
identical with Bacillus coli com-
munis, There are seven varieties
of this bacillus. They were iso-
lated from cases of infantile diar-
rhoea.
Bacillus in infantile diarrhea
(Lesage).—Rods 2-4 » in length,
“75 w in width, and filaments.
517
Colonies irregular ia contour,
colouring the gelatine green.
On the surface of agar they form
a greenish growth, and the gelatine
is coloured green.
Injected intravenously ina rabbit
they produced diarrhea.
They are said to be identical with
Bacillus fluorescens liquefaciens
Bacillus in intestinal diph-
theria in rabbits (Ribbert)—
Rods 3 to 4 » in length, 1 to 14 p
‘in width ; singly, in pairs, and in
filaments.
Colonies greyish ; granular.
They produce in gelatine a deli-
cate growth in the track of the
needle. They are pathogenic.
They were isolated from the
intestine of rabbits suffering from
.a diphtheritic inflammation of the
mucous membrane.
Bacillus in jequirity infusion
(see Bacillus of Sattler).
Bacillus in measles (p. 283).
Bacillus in noma (Schimmel-
busch).—Rods singly, in pairs, and
filaments.
Colonies circular, greyish-white,
granular, with irregular margins.
In the depth of gelatine they
produce agranular filament anda
patch on the surface.
On agar and potato the growth
is greyish-white.
They are pyogenic in rabbits.
They were inoculated from a
case of noma.
Bacillus in ophthalmia (p.190).
Bacillus in potato rot.—Rods
25 to 4 » in length, ‘7 to 8 mw in
width ; singly, in chains, and in fila-
ments. Spore-formation present.
The bacilli inoculated in the
depth of gelatine produce a funnel-
shaped area of liquefaction.
On agar the growth is composed
of greyish-white slimy colonies.
They were isolated from diseased
potatoes. ;
Bacillus in purpura hemor-
rhagica (Tizzoni and Giovannini).
—Rods °75 to 1°3 p in length, ‘2 to
‘4 » in width, singly, in pairs, and
in masses.
Colonies have a greyish-yellow
nucleus and a marginal zone of fine
518
filaments both in gelatine and agar.
Cultures produce a disagreeable
odour.
Subcutaneousinjectionsin guinea-
pigs and rabbits produce local oedema
and death, with hemorrhages in the
internal organs.
They were isolated from the blood
in fatal cases of purpura in children.
Bacillus in putrid bronchitis
(Lumnitzer).—Rods 1°5 to 2 » in
length, slightly curved. They can
be cultivated at 37°C. Thecolonies
on agar are greyish-white.
The bacilli inoculated on blood
serum produce colonies which co-
alesce and form a greyish-white
film. Cultures have a disagreeable
odour.
Injected into the lungs of rabbits
they produce purulentinflammation.
They were isolated from the
sputum in cases of putrid bronchitis.
Bacillus in “red-cod” (Dautec).
—Rods similar to Bacillus tetani,
with terminal spore-formation.
S Colonies are circular ; reddish.
,On the surface of obliquely
solidified gelatine they form a red
gtowth in the track of the needle
slowly followed by liquefaction.
Cultivated on dried cod they
produce a red colour.
They were isolated from red-cod.
oo in rhinoscleroma (p.
Bacillus in saliva (Fiocca).—
Very short rods °2 to ‘33 p in width.
Colonies circular, granular, and
yellowish.
On the surface of obliquely
solidified gelatine they form a
growth composed of transparent
droplets.
On potato they form a transparent
film.
In broth flocculi appear.
They are pathogenic in rabbits
and other small animals, and they
are probably a variety of the bacillus
of hemorrhagic septicemia.
They were isolated from saliva of
cats and dogs.
Bacillus in whooping cough
(Afanassiew).--Rods °6 to 2-2 win
length, singly, in pairs, and short
chains.
DESCRIPTION OF SPECIES.
Colonies granular, brownish.
Inoculated in the depth of gela-
tine there is a scanty growth in the
track of the needle and a greyish
growth on the free surface.
On agar the growth is greyish.
On potato the growth is shining
and yellowish or brownish.
They are said to produce symp-
toms in rabbits and dogs compar-
able to those of whooping cough.
They were isolated from the
throat in cases of whooping cough.
Bacillus incanus (Pohl).—Rods
1-2 » in length, ‘8 » in width.
They produce rapid liquefaction
in the track of the needle when
inoculated in the depth of gelatine.
On agar a thick white growth
develops.
On potato a whitish growth
spreads over the surface.
They were isolated from the
water of marshes.
Bacillus indicus (Koch).—Very
short rods with rounded ends.
Fic. 204.—Bacittus Innicus CoLoNiEs
In Nutrient AGAR, x 60.
The colonies have a scarlet tint.
They are round, ovoid, or spindle-
shaped, and have granular margins.
In the track of the needle beneath
the surface no pigment is formed.
Cultivated in nutrient gelatine
they liquefy it and colour it crimson,
and the growth of a darker crimson
hue subsides to the bottom of the
tube.
On the surface of nutrient agar-
agar the appearances are very
characteristic. In a pure cultiva-
tion a brilliant vermilion-coloured
reticulated pellicle develops on the
surface. (Plate IT. Fig. 3.)
DESCRIPTION OF SPECIES.
They form a vermilion layer on
potato,
They were isolated by Koch in
India from the intestinal contents
of an ape.
Bacillus indigogenes (Alvarez).
Rods 3 p» in length and 1°5 p in
width, singly, and in chains ; capsu-
lated.
On agar they produce a yellowish-
white layer, and are said to develop
an indigo-blue colour in infusions
of leaves of the indigo plant.
Intravenous injections in guinea-
pigs are said to produce death in a
few hours.
They were isolated from the
leaves of the indigo plant.
Bacillus indigonaceus (Clis-
sen).—Rods 1°6 to 3 u long, ‘8 to
‘9 » wide ; non-motile.
They form a sky-blue layer on
the surface of gelatine.
On potato the growth is dark-
blue, and later hasa metallic lustre.
Bacillus indigoferus, which was
found in water at Kiel, is only
to be distinguished by its motility.
Bacillus inflatus (A. Koch).—
Rods 4°6 to 5°5 » in length, °6 to
18 » in width, and filaments.
The colonies send out delicate
processes.
The bacilli inoculated in the
depth of gelatine send out fine
filaments in the track of the needle
followed by slow liquefaction.
On agar they form a shining
brownish layer.
In broth a pellicle forms on the
surface,
They occur in the air.
Bacillus inunctus (Pohl).—
Rods 3°5 » in length, °8 to 9 » in
width.
Inoculated in the depth of gela-
tine they grow both in the track of
the needle and on the surface ;
liquefaction follows in time.
On agar they form a whitish
growth.
They were isolated from the
water of marshes.
Bacillus invisibilis (Vaughan).
—Rods ; motile.
The colonies are irregular and
yellowish.
519
Inoculated in the depth of gela-
tine they grow both in the track of
the needle and on the surface.
On agar they form a white
growth.
On potato they develop an in-
visible layer.
They occur in water.
Bacillus iridescens (Tataroff).-—
Rods from 3:5 to 52 » in length
and threads. Spore-formation pre-
sent ; slightly motile.
The colonies have a characteristic
appearance recalling that of the
convolutions of the brain.
Inoculated in the depth of gela-
tine there is a depression at the
point of puncture, and a thread-
like growth along the needle
track.
On agar they form a thick, un-
even, moist, greenish-yellow, irides-
cent growth, with a pitted surface.
Blood serum is liquefied.
On potato there is a dry, thick,
dark yellow growth like honey.
Broth is rendered turbid, and
there is a yellow deposit.
They are found in water.
Bacillus lactis aerogenes (Es-
cherich).—Rods short and thick,
‘5 to ‘8 » broad, and 1 to 2 pu long,
with rounded ends ; usually in pairs
side by side and also in irregular
heaps. Non-motile; spore-forma-
tion not observed. They grow best
at 37°C.
Colonies on the surface are raised,
moist, shining and porcelain-white.
Below the surface they have a
yellowish nucleus.
Inoculated in the depth of gela-
tine the rods form an abundant
nail-shaped growth. On potato the
culture is composed of white colo-
nies, and bubbles are formed. The
colonies may coalesce and produce
a creamy layer.
On blood serum there is a raised,
moist, shining, white growth.
In milk sugar or grape sugar
solutions they produce gas.
Injected subcutaneously in rab-
bits and guinea-pigs they cause
death in from one to three days,
and the bacilli are found in the
blood and internal organs.
520
They were found in the intestinal
tract of animals fed with milk and
of infants at the breast.
Bacillus lactis albus (Léffler).
—Rods, 3:4 » in length, 96 p in.
width, and filaments. Spore-forma-
tion present.
Inoculated in the depth of gela-
tine they slowly liquefy the upper
part, and a white scum forms on the
surface.
On agar they form a white layer.
On potato the growth is dry and
white. They coagulate milk.
They occur in milk.
Bacillus lactis erythrogenes
(Hueppe).—Short rods, 1 to 1-4 pu
in length and 3 to ‘5 » in width,
and filaments. Colonies small and
circular ; greyish-white ; later yel-
low and surrounded by liquefied
gelatine with a pink tinge.
In the depth of gelatine the
growth in the track of the needle
is scanty, but on the surface a
whitish patch forms which after-
wards turns yellow, and the gela-
tine is coloured pink. Later lique-
faction sets in, and the liquefied
gelatine is turbid and pink.
On agar a shining yellow layer
develops, and the same on potato.
In broth the bacilli produce tur-
bidity, and they coagulate milk.
They occur in “red milk.”
Bacillus lactis pituitosi (Lof-
er).—Rods slightly bent.
Colonies circular, greyish-white.
On agar and potato they pro-
duce a greyish-white layer.
They render milk viscid.
They occur in milk.
‘Bacillus latericeus (Adametz
and Eisenberg).—Rods and _fila-
ments.
Colonies circular, granular, red-
dish-brown.
In the depth of gelatine there is
a scanty growth along the track
of the needle and a_brick-red
growth on the surface.
On potato the growth is also
brick-red.
They occur in water.
Bacillus leporis lethalis (Gibier
and Sternberg).—Rods 1 to 3 » in
length, ‘5 » in width.
DESCRIPTION OF SPECIES.
Colonies transparent and with
the appearance of broken glass.
In the depth of gelatine there
is a growth along the track of the
needle with a conical area of lique-
| faction at the upper part, and a
white sediment.
On agar they form a_ trans-
lucent film. They liquefy blood
serum.
On potato the growth is pale-
yellow.
Cultures injected into the peri-
| toneal cavity of rabbits are toxic.
They were isolated from the in-
testinal contents in cases of yellow
fever.
Bacillus leprz (p. 407).
Bacillus leptosporus (L. Klein).
—Rods_ resembling _ hay-bacilli,
singly, in chains and long twisted
filaments.
The spore-membrane is said to
form part of the newly grown
bacillus, and the filaments are de-
scribed as possessing peculiar spas-
modic movements.
They were isolated from a con-
taminated culture. f
Bacilius limbatus acidi lactici
(Marpmann).—Rods short, thick ;
singly, in pairs ; capsulated.
Colonies white.
In the depth of gelatine they
develop slightly in the track of the
needle, and produce a white patch
on the free surface.
In milk they produce coagula-
tion and lactic acid.
They occur in milk,
Bacillus limosus (Russell).—
Rods 3 to 4 in length, 1:25 » in
width ; singly, in pairs and chains ;
spore-formation present.
Colonies transparent, surrounded
by filamentous processes.
In the depth of gelatine pre-
pared with sea-water, liquefaction
occurs rapidly in the track of the
needle, and a deposit forms at
the bottom and a thin skin on the
surface.
On agar they form a white layer,
and in broth turbidity and a thick
scum.
On potato the growth is greyish-
white.
DESCRIPTION OF SPECIES.
They were isolated from deep-
sea dredgings.
Bacillus liodermos (Fliigge). —
Small short rods with rounded
ends ; actively motile.
Colonies with irregular outlines
float on liquefied gelatine in the
form of small white flakes.
Inoculated in gelatine a greyish
growth occurs along the track of the
needle, but the medium later be-
comes liquefied and a greyish-white
floceulent deposit settles at the
bottom.
On potato a smooth shining yel-
lowish-white layer spreads quickly
over the whole surface, and after
some days becomes opaque and
slightly wrinkled.
They occur on potato.
Bacillus liquefaciens (Hisen-
berg).—Rods short and thick, with
rounded ends. Very motile.
Colonies round, with smooth
edges and slimy centres. Lique-
faction follows, and a putrefactive
odour is noticed.
In gelatine they make a funnel-
shaped whitish growth along the
track of the needle.
On potato the growth is pale
yellow.
They occur in water.
Bacillus liquefaciens commu-
nis (Sternberg).—Rods 1 to 2» in
length, and “7 » in width; singly
and in pairs.
In the depth of gelatine they
produce rapid liquefaction in the
track of the needle.
On potato a wrinkled pinkish
layer is formed.
They were isolated from the eva-
cuations of yellow-fever patients.
Bacillus liquefaciens magnus
(Liideritz).—_Rods 3 to 6 pw in
length, -8 to 1:1 p in width, and
filaments. They are anaerobic.
Colonies develop below the sur-
face of the gelatine, and liquefaction
extends upwards to the surface.
The bacilli inoculated in the
depth of gelatine cause liquefaction
in the lower part of the track of
the needle.
In the depth of agar the colonies
have delicate branches.
521
They liquefy blood-serum, and
produce a putrefactive odour.
They occur in earth.
Bacillus liquefaciens parvus
(Liideritz).— Rods 2 to 5 » in
length, 5 to ‘7 w in width, and
filaments. They are anaerobic.
Colonies are white, and liquefy
gelatine ; but in agar they are
spherical or almond-shaped.
In the depth of gelatine isolated
colonies appear in the track of the
needle, and in the depth of agar
there is gas formation.
They occur in earth.
Bacillus liquidus (Frankland).
—Rods short and flat with rounded
ends, usually in pairs, the length
of each pair varying from 1:5 » to
3°5 p. They are very variable in
size ; highly motile; spore-forma-
tion not observed.
Colonies form cup-shaped excava-
tions, with almost clear, colourless
contents. The edges are at first
smooth and circular, but they
become serrated and granular, and
soon coalesce.
A broad funnel-shaped depression
forms along the whole track of the
needle, containing turbid liquid and
masses of flocculent material. Later
a thin pellicle forms on the surface,
which sinks if the tube is shaken.
On agar they grow quickly, form-
ing a smooth shining layer.
On potato a thick flesh-coloured
growth appears.
Broth is rendered turbid with an
abundant sediment, and after a few
days a pellicle forms.
They are common in unfiltered
water.
Bacillus litoralis (Russell).—
Colonies granular, with regular
contour ; slowly liquefying.
In the depth of gelatine they
develop a growth in the track of
the needle, and at the upper part
produce a cup-shaped cavity lined
with the culture. The gelatine is
tinged with brown in the vicinity.
On agar they produce a greyish-
white film, and in broth turbidity.
Inoculated in the depth of
gelatine the bacilli form a
tunnel-shaped liquefaction along
522 DESCRIPTION
the track of the needle, and the
whole of the gelatine gradually
becomes liquid, with a flocculent
deposit at the bottom and a greyish
wrinkled stain on the surface.
On potato a thick wrinkled whitish
skin forms, which rapidly grows
over the whole surface. On at-
tempting to raise this skin it will
be found to be attached to the
potato by a mucous substance
which may be drawn out in long
threads. According to Hueppe the
bacilli cannot form any ropy sub-
stances from sugar, but they have
an energetic diastatic action. They
coagulate the casein in milk in a
similar manner to rennet.
The bacilli are ubiquitous.
Bacillus lividus (Plagge and
Proskauer ).—Rods.
Colonies blue-black, liquefying.
In the depth of gelatine they
produce a colourless thread in the
track of the needle and a violet
layer on the surface followed by
gradual liquefaction.
On agar the growth is blue-black,
and on potato violet.
They were isolated from water.
They are probably identical with
Bacillus ianthinus, or merely a
luteus
variety.
Bacillus (Fligge). —
Short immotile rods.
Colonies irregular in form, appear
brownish under a low power, and
yellow to the naked eye.
In test-tube cultivations they
form a yellow growth without
liquefying the gelatine.
They occur contaminating plate-
cultivations.
Bacillus maidis (Cuboni). —
Rods 2 to 3 p» in length, singly,
in pairs, and in chains; spore-
formation present.
Colonies granular, with wrinkled
periphery ; later, liquefying.
In the depth of gelatine they
produce rapid liquefaction in the
track of the needle.
On agar a dry wrinkled white
film spreads over the surface.
On potato the growth is wrinkled,
and later yellowish-brown. They
liquefy blood serum.
OF SPECIES.
They were isolated from human
evacuations and infusions of maize.
Bacillus mallei (p. 452).
Bacillus megatherium (De
Bary).—Rods 2°5 » wide and four
to six times as long, with rounded
ends and slightly curved, and in
short irregular chains.. Transverse
division occurs, each segment
attaining the length of the original
rod. In the fresh state they appear
non-articulated, but when treated
Fic. 205.—Bacittus M&cATHERIUM.
(a) A chain of rods x 250, the rest
x 600. (b) Two active rods: d and f,
successive stages of germination; h
and 1, successive stages of germina-
tion. (De Bary.)
with a dehydrating agent they are
seen to be composed of short seg-
ments with granular contents. They
are motile.
Colonies are small and circular,
and the gelatine is liquefied.
In the depth of gelatine the
bacilli grow rapidly, forming a
funnel-shaped liquefaction in the
upper part.
On agar they form a whitish
layer on the surface, and the jelly
acquires a dark colour.
On potato yellowish-white cheesy
colonies are formed round the point
of inoculation. In cultures there
is copious spore-formation. They
grow best at 20° C.
They were isolated originally
from boiled cabbage.
DESCRIPTION
Fic. 206.—PuRE-CULTURE OF BACILLUS
MEGATHERIUM IN GELATINE,
Bacillus membranaceus ame-
thystinus (Hisenberg).—Short rods
with rounded ends from 1 to 1:4 p
long, and ‘5 to ‘8 » broad. They
are grouped together irregularly.
Some individual bacilli stain more
deeply at the ends than in the
middle. Non-motile. They grow
only between 15° and 20° C. Spore-
formation uncertain.
The colonies gradually assume a
violet hue, and after liquefying the
gelatine float on the surface as violet
pellicles, resembling a membrane
stained with gentian violet.
Inoculated in the depth of
gelatine a yellowish-white growth
appears on the free surface, which
after ten days or more becomes
violet. lL.iquefaction takes place
gradually, and in about a month
a thick violet layer covers the gela-
tine which lies beneath the liquid
part.
On agar the growth, which at first
has a yellowish milky appearance,
becomes violet after eight or ten
days. In three or four weeks it
has become very much wrinkled,
and has a beautiful deep-violet
colour with a metallic lustre. The
OF SPECIES. 323
| jelly is not stained, and the growth
can be easily removed from its
surface.
On potato they grow slowly, and
form a dirty yellow or olive-green
colour.
In broth they grow very slowly ;
after some weeks a violet deposit
and pellicle are formed, and the
liquid between becomes dark brown.
They were found in well water.
Bacillus meningitidis puru-
lentae (Neumann and Schaffer).—
Rods 2 » in length, 6 to ‘7 » in
width, and filaments.
Colonies granular, greyish.
In the depth of gelatine a greyish-
yellow filament develops, composed
of closely packed colonies, and on
the surface a greyish layer.
On potato the growth is moist
and white.
They are pyogenic in small ani-
mals and dogs.
They were isolated from a case
of purulent meningitis.
Bacillus mesentericus fuscus
(Fliigge).—Rods small and short,
singly, in chains of two and four.
Actively motile. Spore-formation
present.
Colonies are at first roundish and
rather white, with a sharp outline ;
later delicate brownish-yellow pro-
cesses appear. Liquefaction occurs
rapidly.
Inoculated in the depth of gela-
tine a whitish growth forms along
the track of the needle, the upper
portion of which soon liquefies ;
greyish flakes float in the liquefied
portion.
On potato a smooth yellowish
growth appears on the first day,
but it soon becomes brown and
wrinkled. It remains relatively
thin and superficial, and quickly
spreads over the whole surface.
They are found in hay dust, in
the air, on the surface of potatoes,
and are very widely distributed.
Bacillus mesentericus ruber.—
Slender rods, singly, in pairs, and
in filaments.
Colonies are circular and yellow-
ish until they come to the surface,
when they produce a network and
524 DESCRIPTION
liquefy the gelatine. The network
disappears and a little deposit occurs
at the bottom of the liquefied
area..,
Inoculated in the depth of gela-
tine a whitish cloudy growth forms
along the needle track, liquefaction
sets in and extends-until the gela-
tine is completely liquefied.
On potato a thin crinkled film is
formed, which is yellowish or red-
dish-yellow in colour.
They occur on potato.
Bacillus mesentericus vulga-
tus (Fliigge).—Rods large and
thick, often forming pseudo-threads.
They have an oscillating movement.
Spore-formation present.
The colonies are bluish-white and
almost transparent, though the
centres become gradually opaque.
They sink in the liquefied gelatine,
and are granular with irregular
contour.
Bacillus multipediculus
(Fliigge).—Rods long and slender.
Non-motile.
The colonies consist of a central
oval nucleus, from which numerous
tapering processes shoot out mostly
towards one pole. This form of
growth gives a curious resemblance
to an insect with feet and antenne.
Inoculated in the depth of gela-
tine a whitish line forms along the
track of the needle, from which
short processes grow out.
On potato a rather scanty dirty-
yellow growth forms, and the sur-
face of the potato becomes dis-
coloured around it.
They are often found as a con-
tamination on potato.
Bacillus muscoides (Liborius)
—Rods 1 p thick, sometimes form-
ing threads; slightly motile, and
with round or oval spores at one
end, They are anaerobic.
The colonies ramify and resemble
a délicate moss.
They were found in the cedema-
tous fluid of field mice inoculated
with garden earth and stale cheese.
Bacillus mycoides (Fligge).—
Rods rather thick, nearly the size
of Bacillus anthracis. Motile,
often forming long pseudo-threads,
OF SPECIES.
Oval and highly refractive spores
both in the rods and threads.
Colonies consist of a whitish tur-
bidity in which colourless branched
and interwoven processes are seen ;
these increase rapidly, and after
twelve to twenty hours appear like
the mycelium of a fungus.
Inoculated in the depth of gela-
tine they form very fine and closely
set hairs extending from the track
of the needle. Later liquefaction
occurs.
On potato a whitish layer gradu-
ally extends over the surface.
They occur in earth from the
surface of cultivated ground.
Bacillus mycoides roseus
(Scholl).—Rods.
Colonies composed of interlacing
filaments. :
Inoculated in the depth of gela-
tine they produce liquefaction in
the track of the needle ; a reddish
scum forms on the surface, and a
reddish deposit at the bottom of
the liquefied area.
On agar they produce, in the
absence of light, a pink growth.
Bacillus neapolitanus (Emme-
rich).—Short rods °9 » in width.
Fic. 207.—Bacittus NEAPOLITANUS, x
700 (EMMERICH). «, From intes-
tinal contents in a case of cholera ;
6, From_ peritoneal fluid of an
inoculated guinea-pig.
DESCRIPTION OF SPECIES.
Colonies circular, later irregular,
granular, strongly refractive and
yellowish - brown. They are
probably identical with Bacillus coli
communis.
They were isolated from cases of
cholera at Naples.
Bacillus necrophorus (Léffer).
—Rods and filaments.
They cannot be cultivated on the
ordinary media. In rabbit broth
they give rise to fluffy masses of
filaments.
Intravenous injection produced
in rabbits a pyemic condition in
about a week. The bacilli were
found in the pus.
They were isolated from a rabbit
which had been inoculated with
fragments of a condyloma.
Bacillus nitrificans (Wino-
gradsky).—Very small rods ‘5 » in
in length, singly and in zoogloea.
Colonies in silica jelly are
lenticular, and sub-cultures in liquid
media produce a gelatinous de-
posit. They are powerful oxidising
agents.
They were isolated from the soil.
Bacillus nodosus parvus
(Lustgarten).—Rods 1:2 to 2°4 » in
length, -4 » in width ; singly and
in pairs.
Inoculated in the depth of agar
they produce a white filament in
the track of the needle composed
of crowded colonies, and on the
surface a hemispherical glistening
growth.
They were isolated from the
human urethra.
Bacillus nubilus (Frankland).—
Slender rods 3 » long and ‘3 » wide,
and threads. Single bacilli have an
active rotatory movement, but the
long threads in broth cultures are
quite motionless. Spore-formation
not observed. :
The colonies appear as cloudy
undefined patches, which rapidly
liquefy the gelatine. They consist
of a tangled mass of threads.
Inoculated in the depth of gela-
tine they produce along the track
of the needle horizontal circular
plates, with a delicate cloud-like
appearance, and liquefaction at the
525
upper part. Later the whole of
the gelatine is liquefied.
On agar they form a thin opales-
cent blue-violet film, the edges of
which exhibit later a distinct violet
fluorescence.
On potato there is a slightly
yellow growth- which is scarcely
visible.
Broth is rendered turbid with a
dirty-white deposit, the surface
being covered by a thin pellicle.
They occur in water.
Bacillus ochraceus (Zimmer-
mann).—Rods 1:25 to 45 » in
length; °65 to -75 » in width;
singly, in pairs, chains, and fila-
ments ; capsulated. ;
Colonies circular, granular, yel-'
low, liquefying.
Inoculated in the depth of gela-:
tine they produce liquefaction in
the track of the needle, and a
yellow deposit.
On agar and potato the growth
is yellow ochre in colour.
They occur in water.
Bacillus edematis aerobicus
(Klein).—Rods ‘8 to 2°4 «in length,
‘7 » wide, and long filaments.
Colonies greyish, transparent,
with irregular contour.
In the depth of gelatine a fila-
ment occurs in the track of the
needle, and gas bubbles in isolated
colonies in its lower part, and a
transparent patch with irregular
margin on the free surface.
On the surface of agar they pro-
duce a greyish-white layer.
In broth there is turbidity with
flocculi.
On potato the growth is yellowish
and viscid.
They give rise to extensive oedema
in guinea-pigs, and in a less marked
form in rabbits.
They occur in earth.
Bacillus cdematis
p. 220).
Bacillus of Belfanti and Pas-
carola.—Very short rods.
Colonies circular, granular, yel
lowish-grey.
Inoculated in the depth of gela-
tine they produce a filament com-
posed of closely-packed minute
maligni
526 DESCRIPTION
colonies, and on the surface a
greyish film.
On agar they produce a greyish-
white growth.
On potato a transparent whitish
film.
They are fatal to rabbits, guinea-
pigs and small birds.
They are probably identical with
Bacillus septiceemize heemorrhagicee.
They were isolated from pus in
a case of tetanus.
They were isolated from deep-sea
dredgings.
Bacillus of Colomiatti—Minute
rods. Spore-formation occurs at
the ends of the rods. They can
be cultivated at 37° C.
They form a thin film on agar
and on blood serum.
They were isolated in cases of
conjunctivitis.
Bacillus of Fulles, No. I.—Rods
1 to 1:2 » in length, °6 » in width.
Colonies circular, granular, yel-
lowish-brown.
On the surface of gelatine they
produce a thin film, and in broth
turbidity and flocculi.
On potato the growth is yel-
lowish.
No. II. Very short rods.
Colonies circular, granular, yel-
lowish.
In the depth of gelatine the
growth resembles Friedlinder’s
pneumococcus.
On potato the growth is yellowish.
They were isolated from earth.
Bacillus of Guillebeau.—No. I.
Short rods 1 to 2 » in length, 1p
in width.
Colonies spherical, granular.
In gelatine the bacilli produce a
growth in the track of the needle
and a white patch on the surface.
On agar the growth is white, and
on potato yellowish, viscid, and con-
taining gas bubbles.
They coagulate milk.
No. II. Rods resembling the
above described but distinguished
by the production of viscid colonies
and, extremely slowly, of liquefac-
tion in the jelly. p
No. ITI. Rods also resembling the
above mentioned, but colonies are
OF SPECIES.
adherent to the jelly and coarsely
granular.
Milk and other liquid culture
media are rendered extremely vis-
cid.
Bacillus of Letzerich.—Rods
sometimes bent, and filaments.
They rapidly liquefy gelatine.
They produce purulent peritonitis
and death in rabbits.
They were isolated from urine.
Bacillus of Martinez (Stern-
berg).—Short rods 1 to 12 » in
length and 5 to ‘8 » in width;
non-motile.
Colonies circular and translucent,
with a central, nipple-like projec-
tion, and the surface covered with
mosaic markings.
In the depth of gelatine the
growth consists of large spherical
translucent colonies in the track of
the needle, and a thin, translucent,
scanty growth upon the surface.
They were isolated from the liver
in a fatal case of yellow fever.
Bacillus of Nocard. (Vide Strep-
tothrix farcinica.)
Bacillus of Okada.—Short rods
rather thicker than the bacilli of
mouse-septicemia, singly, in pairs
and in filaments. Spore-formation
not observed.
Colonies granular and brownish.
Inoculated in the depth of gelatine
they form a white filament, and on
the surface a milk-white patch.
Inoculated on agar the growth
spreads over the surface forming a
milk-white layer.
In broth they produce cloudiness
and a layer floating on the surface.
They do not grow on potato.
Cultures produce death in mice,
guinea-pigs and rabbits in twenty
hours.
They were isolated from dust.
Bacillus of Roth—No. 1 and
No. 2. Rods.
Two varieties were isolated from
old rags. They appear to be
varieties of Bacillus coli communis.
Bacillus of Sattler.—2 to 4°5 p
long and ‘58 p thick.
They can be cultivated on nutrient
gelatine and blood serum.
Anfusion of jequirity containing
DESCRIPTION OF SPECIES.
the bacilli, inoculated into the con-
junctiva of healthy rabbits, produces
severe ophthalmia. The poisonous
principle is a chemical ferment
abrin. Boiling, which does not
destroy the spores of the bacillus,
destroys the ferment, and cultiva-
tions started from these spores,
though teeming with jequirity
bacilli, are quite harmless (Klein).
The bacilli occur in infusions
of the beans of Abrus precatorius
or jequirity.
Bacillus of Schaffer (Freuden-
reich).—Rods 2 to 3 » in length,
1, in width, and long filaments.
Colonies circular, granular, yel-
lowish.
In the depth of gelatine a growth
develops in the track of the needle
and a greyish layer on the surface.
On agar the growth is greyish
and sometimes brownish, and on
potato yellowish.
In broth with peptone and milk
sugar there is copious formation of
gas-bubbles.
They closely resemble Bacillus
coli communis.
They were isolated from cheese
and potato.
Bacillus of Scheurlen.— Rods
1:5 to 2°5 p in length, ‘5 » in width.
They were isolated from cancerous
growths by Scheurlen, and were
later identified with Bacillus epider-
midis.
Bacillus of Schou.—Short rods
and cocci-forms.
Colonies are spherical, opaque,
and granular.
The bacilli inoculated in gelatine
rapidly liquefy it, and a white de-
posit forms at the bottom of the
liquid.
Rabbitsinoculated in the trachea,
or made to inhale pure-cultures, are
said to develop fatal pneumonia.
They were isolated from rabbits
with pneumonia, following section
of the vagi.
Bacillus of
(p. 351).
Bacillus of Tommasoli.—Short
rods from 1 to 1'8 » in length, and
25 to 3 » in width, singly, and in
short chairs.
swine plague
527
Colonies grey and shiny.
In the depth of gelatine they’
form a filament composed of closely-
packed colonies, and on the surface
a shining mass.
On agar the growth consists of
greyish patches.
On potato the growth is granular
and yellowish-white.
Cultures rubbed into the skin
are said to produce a vesicular
eruption.
They were isolated from the
scalp in a case of sycosis.
Bacillus of Utpadel—Rods 1:25
to 1:5 w in length, and ‘75 to 1 p
in width, singly, in pairs and short
chains.
Colonies milk-white.
On the surface of gelatine the
growth is milk-white, and on agar
yellowish-white.
Injected subcutaneously in cats,
guinea-pigs and mice, they produce
extensive cedema, and a fatal ter-
mination.
They were isolated from the
human intestine.
Bacillus of Winogradsky. (See
Bacillus nitrificans.)
Bacillus ovatus minutissi-
mus (Unna).— Short rods with
pointed ends °6 to ‘8 » in length,
‘4 » in width, singly, and in
masses.
Colonies are minute, granular and
yellowish.
The bacilli inoculated in the
depth of gelatine form a filament
of closely packed greyish-white
colonies, and on the free surface
there is a shiny, greyish-white
layer.
On agar the growth is very simi-
lar, and on potato also.
They were isolated from the skin
in eczema seborrhceicum.
Bacillus oxytocus spernicions
(Wyssokowitch).—Rods short and
thick.
Colonies circular, granular, yel-
lowish, or yellowish-brown.
The bacilli inoculated in the
depth of gelatine produce a growth
resembling Friedlinder’s pneumo-
coccus.
They coagulate milk.
528 DESCRIPTION
The products injected intrave-
nously produce death in from three
to twenty-four hours.
They occur in sour milk.
Bacillus pestifer (Frankland).
—Rods 2:3 pw in length, 1 » in
width, and filaments. Motile.
Colonies resemble those of Bacil-
lus vermicularis.
On agar they produce a dentated
transparent layer, and on potato a
flesh-coloured growth.
They occur in the air.
Bacillus phosphorescens geli-
dus (Forster).—Very short rods.
Colonies circular, granular, yel-
lowish or greenish.
The bacilli, inoculated in the depth
of gelatine, produce very little
growth in the track of the needle,
and a white film on the surface.
On agar and potato the growth
is whitish.
Cultures are photogenic.
They were isolated from phos-
phorescent fish.
Bacillus phosphorescens Indi-
cus (Fischer ).—Rods singly and in
pairs, and filaments. Motile.
Colonies circular, well-defined,
greenish.
The bacilli, inoculated in the
depth of gelatine, produce a greyish
filament in the track of the needle,
and a hemispherical excavation of
the jelly at the upper part. Later
the jelly is liquefied, and there is a
yellowish scum on the surface.
On agar and potato the growth
is white.
Cultures are photogenic.
They were isolated from sea-
water.
Bacillus phosphorescens indi-
genus (Fischer).—Rods 1:3 to 1:2
# in length, ‘4 to ‘7 » in width,
singly, in pairs, and filaments.
Colonies circular, greenish, and
later yellowish.
The bacilli, inoculated in the
depth of gelatine, produce a conical
excavation in the upper part of the
needle track without liquid con-
tents, but with a dry growth on
the sides.
There is no growth on potato.
Cultures are photogenic.
OF SPECIES.
They occur in sea-water and on
phosphorescent fish.
Bacillus plicatus (Zimmer-
mann).—Minute rods, singly, in
pairs, and in short chains.
Colonies yellowish-white.
The bacilli, inoculated in the
depth of gelatine, form minute
isolated colonies, and on the surface
a wrinkled patch with gradual lique-
faction. ;
On potato the growth is dry and
yellowish.
They occur in water.
Bacillus pneumosepticus
(Babés).—Short rods, ‘2 «in width.
Colonies irregular, semi - trans-
parent.
In gelatine, the bacilli grow in
the track of the needle. On agar
the growth is whitish and shining.
Rabbits, guinea-pigs and mice
die in two or three days of septi-
cemia when a culture is injected
subcutaneously.
They were isolated from a fatal
case of septic, pneumonia.
Bacillus polypiformis (Libo-
rius).—Slender rods, spore-forma-
tion present. They are anaerobic.
Colonies composed of peculiar
convoluted processes.
In the depth of blood serum they
produce a cloudiness at the lower
part of the needle track.
They occur in soil.
Bacillus prodigiosus (Jficro-
coccus prodigiosus : Cohn.— Blood
rain, Bleeding host). Very short
rods with rounded ends, and thread
forms ‘5 tol » in width, forming
at first rose-red and then blood-red
zoogloea.
They liquefy gelatine.
They grow luxuriantly on the
sloping surface of nutrient agar-
agar, and on sterilised potato, and
the colour varies from blood-red to
bright-red with sometimes a metal-
lic lustre. The cells themselves
are colourless. The colouring-matter
resembles fuchsine ; it is insoluble
in water but soluble in alcohol.
The addition of acids changes it to
carmine red, and of alkalies to a
yellow colour.
They appear occasionally on
DESCRIPTION OF SPECIES.
bread, boiled rice, and starch paste,
and more rarely on boiled white
of egg and meat. Milk sometimes
becomes coloured blood-red by the
growth of this fungus, an appear-
ance formerly attributed to a disease
of the cow.
‘In Paris in 1843 the micro-
organism was peculiarly prevalent,
attacking especially the bread pro-
duced in the military bakehouses.
Bacillus proteus ‘fluorescens
(Jager).—Short thick rods and
threads. Actively motile.
Colonies resemble minute drops
of water.
The rods inoculated in gelatine
produce a growth similar to that
of Koch’s comma-bacilli.
The jelly becomes greenish, and
a pellicle forms on the surface.
On agar the growth when fully
developed is yellowish-white, with
a green fluorescence.
On potato they form a brown
layer.
They are pathogenic in mice.
They were isolated from the
internal organs of fowls suffering
from an epidemic disease.
( Bacillus pseudo-diphtheriticus
p. ists 5).
Bacillus _pseudo-tuberculosis
(Pfeiffer).—Rods varying in length.
Colonies circular, with dark nu-
cleus and transparent zone.
In the depth of gelatine they
produce a filament composed of
small colonies, and on the surface
a patch with concentric markings.
They grow on agar, but not readily
on potato.
Inoculated in mice, guinea-pigs,
rabbits, and hares, they produce a
fatal result in from six to twenty
days. An abscess forms locally,
the lymphatic glands enlarge and
caseate, and the internal organs
contain nodules resembling tubercle.
They were isolated from the in-
ternal organs of a horse supposed
to be glandered.
Bacillus pulpe pyogenes.—
Rods slightly bent and with pointed
ends ; singly, in pairs, and in chains.
Colonies circular, yellowish-
brown.
529
_ Inoculatedin the depth of gelatine
liquefaction occurs in the upper
part of the needle track and ex-
tends downwards.
Intraperitoneal injection in mice
produces death in from eighteen to
thirty-six hours.
They were isolated from putrid
dental pulp.
Bacillus punctatus (Zimmer-
mann).—Rods 1 to 1-6 in length,
“77 yw In width, singly, in pairs, and
chains.
Colonies composed of stringy
masses in liquefied gelatine.
The bacilli inoculated in the
depth of gelatine produce rapid
liquefaction in the track of the
needle, and a white deposit.
On agar the growth is smooth
and shining.
On potato the growth is brownish.
They occur in water.
Bacillus putrificus coli (Bien-
stock).—Slender, motile rods, 3 »
in length, often less, sometimes
ae
: | {7
ie Ft ¢
i i ee
a)
ae
Fic. 208.—Bacitius Purriricus Cou,
x 1000 (BrENnsTock),
forming long threads.
mation present.
Cultivations in gelatine are iri-
descent.
They are constantly present in
faeces.
Bacillus pyocyaneus (Gessard).
—Slender rods, singly, in twos.
and threes, or in irregular masses.
Spore-formation present. :
White colonies appear in twenty-
four hours, which liquefy the gela-
tine. The whole of the medium
acquires a greenish shimmer.
If the bacilli are cultivated in
gelatine, the jelly is liquefied, and
coloured green byreflected light, and
a deep orange by transmitted light.
On agar they form a white layer,
and colour the medium a pea-green.
34
Spore-for-
530 DESCRIPTION
On potato a dry rust-brown
growth appears at the seat of
Inoculation, which becomes green
when treated with ammonia.
The pigment formed by the micro-
organism is a definite principle—
pyocyanin. It can be extracted
with chloroform from pus and from
washing of bandages; it is soluble
in acidulated water, which it colours
red. In neutral solution it becomes
blue. It crystallises in chloroform
in long needles ; and forms some-
times lamelle and prisms.
They cause death in guinea-pigs
when injected into the abdominal
cavity. Rabbits are not killed by
intravenous injection.
The bacilli are antagonistic to
anthrax bacilli. Charrin and others
have shown that rabbits inoculated
with a pure-culture of Bacillus
pyocyanus after inoculation with
Bacillus anthracis will not succumb
to anthrax. Woodhead and Wood
produced similar results by using
sterilised cultures, showing that the
results were due to the chemical
products of the bacilli.
The rods occur in the pus of those
cases in which the wounds and pus-
stained bandages exhibit a greenish-
blue colour.
Bacillus pyogenes fetidus
(Passet).—Small rods, about 1:45 p
in length, and -58 w in width ;
Fic. 209.—Bactttus PyocEnes
Fartipus, x 790 (PASsEtT).
often in pairs. or linked together
in chains. They are motile, and
‘spore-formation occurs.
Colonies like white points appear
after twenty-four hours, and de-
velop into greyish spots, and these
enlarging coalesce into a layer.
Cultivated in nutrient gelatine
OF SPECIES,
a greyish, veil-like growth forms
on the surface.
In nutrient agar-agar the culti-
vation resembles the growth in
gelatine. On blood serum a moder-
ately thick greyish-white streak
develops, and on sterilised potato
an abundant, shining, brownish
culture.
From all these media a putrid
odour emanates, but no smell is
detected from a cultivation in milk,
Inoculated into mice and guinea-
pigs, abscesses are produced or death
from septicemia results.
They were isolated from putrid
pus.
Bacillus pyogenes soli( Bolton).
—Rods resembling Bacillus diph-
therie.
Colonies granular, faintly yellow.
The bacilli inoculated in the depth
of gelatine form colonies in the
track of the needle.
They are pyogenic in mice and
rabbits.
They are present in earth.
Bacillus radiatus (Luderitz)—
Rods 4 to 7 pw in length, 8 w in
width, and filaments. Motile.
They are anaerobic. Spore-forma-
tion present.
Colonies are composed of delicate
interlacing filaments.
In the depth of gelatine a growth
occurs at the-lower part of the
needle track, from which fine fila-
ments are given off in the sur-
rounding gelatine, and liquefaction
follows. The growth in the depth
of agar is also composed of fine
filaments.
In sub-cultures they produce a
cloudy liquefaction.
Cultures have a peculiar odour.
They occur in earth.
Bacillus radiatus aquatilis
(Zimmermann).—Rods 1 to 6:5 p
in length, °65 » in width.
Colonies white, with a marginal
zone of radiating filaments.
The bacilli inoculated in the
depth of gelatine grow in the track
of the needle, and excavate and
liquefy the surrounding gelatine,
and form on the free surface a
wrinkled patch, which later subsides
DESCRIPTION
to the bottom of the liquefied
area.
On agar a_ smooth slightly
brownish layer is formed, and on
potato it is yellowish.
They occur in water.
Bacillus ramosus (Hisenberg).—
Rods singly, and in chains ; fila-
ments. Spore-formation present.
Colonies are composed of curi-
ously twisted filaments.
The bacilli inoculated in the depth
of gelatine produce delicate fila-
ments extending in all directions
from the track of the needle, fol-
lowed by liquefaction ; later a skin
forms on the surface. There is a
sediment at the bottom of the tube.
On agar they form a greyish
filamentous layer, and on potato
a whitish growth.
They occur in earth and water.
Bacillus reticularis (Jordan).—
Rods 5 pw in length, 1 » in width ;
singly, and in short chains. Motile.
Colonies are composed of radi-
ating filaments, and liquefy the
gelatine, forming an excavation with
a reticulated lining.
The bacilli inoculated in the
depth of gelatine produce filaments
extending from the track of the
needle, and at the upper part the
jelly is excavated in the form of
a cup. On agar they form a dry
layer, and on potato a white woolly
growth.
Broth is made turbid, and milk
slowly coagulated.
They occur in water.
Bacillus rosaceus metalloides
(Bacterium rosaceum metalioides,
Dowdeswell ; Magenta bacillus).—
Rods °6 to ‘8 » in breadth.
Colonies in the depth of gelatine
are colourless, but superficial ones
are prominent and magenta in
colour.
On the surface of obliquely solidi-
fied gelatine they form a beautiful
magenta band with a metallic
lustre. The gelatine is not lique-
fied. Similar growths are obtained
on agar and potato.
It is one of the most striking
of all the chromogenic bacteria.
Cultures have the appearance of
OF SPECIES. 531
having been stained with an alco-
holic solution of fuchsine. The
colour varies in subcultures from
magenta to a sealing-wax-red,
Bacillus rubefaciens (Zimmer-
mann).—Rods ‘75 to 165 pw in
length, °32 » in width, singly, in
pairs, and in chains.
Colonies are faintly reddish-
yellow.
The bacilli inoculated in the
depth of gelatine grow along the
track of the needle and form a
greyish layer on the surface ; later
the jelly acquires a reddish tint.
On agar the growth is grey and
abundant.
On potato the growth is at first
grey, later reddish-brown, and the
surface of the potato has a pink
discoloration.
They occur in water.
Bacillus rubellus (Okada).—
Rods resembling those of malignant
edema. They occur singly, in
pairs, and filaments; are motile,
and possess flagella, and are often
capsulated. They are anaerobic.
Colonies are whitish, with off-
shoots in the surrounding gelatine,
which, later, is liquefied, and has a
reddish tinge.
The bacilli inoculated in the
depth of gelatine produce a growth
in the lower part of the needle
track composed of isolated colonies
with radiating processes. The‘jelly
is liquefied, at first in the part
corresponding with the growth, and
later completely. The liquefied
gelatine is coloured red.
In agar the growth extends from
below upwards, and the jelly is
coloured red.
In broth they grow rapidly.
They were isolated from dust.
Bacillus ruber (Breunig, Bucille
rouge de Kiel, \aurent).—Rods
2:5 to 5 » long, and °7 to ‘8 » broad.
Slightly motile.
Colonies below the surface of
gelatine are pale yellow, and super-
ficial ones are blood-red.
Inoculated in the depth of gela-
tine they liquefy it and colour it
bright red. There is also forma-
tion of gas bubbles.
532 DESCRIPTION
- Potato is rapidly covered with a
purplish-red growth. Broth be-
comes turbid, and pink in colour.
Milk is coagulated, and a blood-
red colour develops on the surface
and gradually extends.
They were found in water.
Bacillus rubescens (Jordan).—
Rods 4 » in length, ‘9 » in width,
singly, in pairs, and short chains.
Colonies pure-white.
Inoculated in the depth of gela-
tine there is a little growth in the
track of the needle, and a pure-
white prominent patch on the free
surface. :
On agar the growth is white and
shining, and later has a pink tinge.
On potato the growth is flesh-
coloured. )
Broth becomes turbid, and a scum
forms on the surface. ;
Milk after a time acquires a pink-
ish colour.
They occur in sewage. : i
Bacillus rubidus (Eisenberg).—
Rods and filaments.
Colonies circular, granular, and
slightly red.
The bacilli inoculated in the depth
of gelatine produce liquefaction and
a brownish-red colour.
On agar and potato they form a
brownish-red growth, and liquefy
blood serum.
They occur in water.
Bacillus sanguinis typhi
(Brannan and Cheeseman).—Rods
1 to 2°5 » in length, ‘5 to ‘8 p in
width ; singly, in pairs, and in
chains, and involution forms.
. Colonies granular, pale-brown.
The bacilli inoculated in the
depth of glycerine-agar produce a |
growth in the track of the needle
composed of isolated, minute, white
colonies.
Rabbits inoculated die in from
two weeks to a month.
They were isolated from the |
blood of patients suffering from
typhus fever.
Bacillus saprogenes (Rosen-
bach).—Three rod-formed organ-
isms have been described by
Rosenbach as intimately associ-
ated with putrefactive processes.
OF SPECIES.
No. 1.—Large rods (Fig. 210),
which form an irregular sinuous
streak with a mucilaginous appear-
ance when cultivated on nutrient
agar-agar. Spore-formation pre-
sent. They grow also very readily
on blood serum, and all cultivations.
‘yield the odour of rotting kitchen
refuse. They are not pathogenic.
No. 2.—Rods shorter and thinner
than No. 1. They develop very
rapidly on agar-agar, forming trans-
parent drops, which become grey.
The cultivations yield a character- -
istic odour similar to the last.
Dy al
o Zz,
‘SI oh
& 4
we Wil
Fig. 210.—BaAcILLUs SAPROGENES, No. 1.
(Rosenbach. )
They are pathogenic in rabbits.
They appear to be identical with
Bacillus foetidus (Bacterium foeti-
dum, Thin). They were isolated
from a patient suffering from pro-
fusely-sweating feet.
No. 3—See Bacterium
genes. ;
Bacillus scissus (Frankland).—
Very short rods, 1 to 2 » in length,
and 1 » in width..
Colonies yellowish, opaque in the
centre, and periphery dentated.
Inoculated in the depth of gela-
tine there is no growth in the
track of the needle, but a shining
layer forms on the surface, and the
jelly is coloured greenish.
On agar the growth is shining
and the jelly coloured green.
On potato the growth is flesh-
coloured.
They occur in earth.
Bacillus septicemie hzemor-
rhagice (p. 231).
Bacillus septicus (Klein).—
Rods varying in size. Non-motile.
They form threads or leptothrix
filaments, and are rounded at the
ends. They are anaerobic, and form
spores independently of access to air.
sapro-
DESCRIPTION OF SPECIES,
In-a nourishing fluid they are
overcome by the presence of micro-
cocci, Bacterium termo or Bacillus
subtilis.
They occur in the soil, in putrid
blood, and many putrid albuminous
fluids, and occasionally in the blood-
vessels of man and animals after
death.
Bacillus septicus acuminatus
(Babés).—Rods with lancet-shaped
ends, about the size of the bacilli
of mouse-septicemia. They exhibit
polar staining. They can be culti-
vated at 37°C.
On agar and blood serum the
colonies are circular, transparent,
and later coalesce and form a yel-
lowish layer.
They are fatal to rabbits and
guinea-pigs in from two to six
days.
They were isolated from an infant
after death from-septic infection
occurring five days after birth.
Bacillus septicus agrigenus
(Nicolaier).—Rods resembling Ba-
cillus septiczemize hemorrhagic.
Colonies circular, granular, with
concentric zones of varying tints of
brown.
Intravenous injections are fatal
to rabbits in twenty-four to thirty-
six hours, and bacilli abound in the
blood.
They occur in recently manured
soil.
Bacillus septicus keratoma-
lacie (Babés).—Short thick rods
singly, and in pairs ; often capsu-
lated.
Colonies white, with dentated
contours.
The bacilli, inoculated in the depth
of gelatine, grow in the track of
the needle and on the surface ; gas
bubbles are developed.
On agar the growth is arbores-
cent and opalescent.
-On blood serum they form a
shining, somewhat transparent,
dentated film. Cultures have an
ammoniacal odour:
They produce purulent inflam-
mation of the cornea.
They were isolated from the
cornea in a case of. septicemia
533
following keratomalacia in a
child.
Bacillus septicus ulceris gan-
grenosi (Babés).—Short rods ‘5 to
6 pin width. ‘
Inoculated in the depth of gela-
tine they produce liquefaction and
gas in the track of the needle.
On agar greyish-white shining
patches are found.
On potato they develop a trans-
parent film.
They are pyogenic in rabbits and
mice.
They were isolated from the
internal organs and blood in a case
of septicemia following gangrene.
Bacillus _septicus vesice
(Clado).—Rods 1°6 to 2 » in length,
‘dD p in width.
Colonies circular, ‘transparent,
yellowish.
The bacilli inoculated in the
depth of gelatine form a delicate
filament composed of closely-packed
colonies, and on the surface there is
a filmy growth.
On agar they form a greyish-
white layer, and on potato the
growth is dry and brown.
They are poisonous to rabbits,
guinea-pigs, and mice.
They were isolated from urine
from a case of cystitis.
Bacillus sessilis (L. Klein).—
Rods resembling those of the hay
bacillus.
They are said to be distinguished
by fission commencing in a newly
formed rod before it has been set
free from the spore.
They were isolated from the blood
of a cow.
Bacillus smaragdino-phospho-
rescens (Katz).—Rods with pointed
ends, 2 » in length, 1 » in width.
Colonies circular, faintly yellow,
with concentric rings.
Inoculated in the depth of gela-
tine a white filament forms in the
track of the needle and a greyish-
white patch on the free surface ;
and there is sometimes liquefaction.
In broth they produce turbidity.
On potato they produce a thin
brownish-yellow film.
They are photogenic, and: the
534 DESCRIPTION
phosphorescence is most marked in
cultures containing an excess of
salt.
They were isolated from a phos-
phorescent herring.
Bacillus smaragdinus fotidus
(Reimann).—Slender rods slightly
bent.
Colonies on agar irregular, with
a yellowish granular nucleus, and
transparent marginal zone.
The bacilli inoculated in the
depth of gelatine produce a growth
in the track of the needle, and
liquefaction at its upper part,
and a greenish coloration.
In the depth of agar the medium
is coloured green.
On potato the growth is brown.
Cultures emit a strong odour.
Intravenous injection produces
death in rabbits in forty-eight
hours.
They were isolated from nasal
mucus in ozzena.
Bacillus solidus (Liideritz).—
Rods 1 to 10 p» in length, °5 » in
width.
They are anaerobic. Cultivated in
grape-sugar gelatine they produce
gas bubbles and a penetrating foul
odour. The colonies‘ are spherical,
and in agar under a low power
are seen to be composed of fine
filaments like cotton wool.
In broth with exclusion of oxygen
they produce a copious growth with
abundant formation of foetid gas.
They were isolated from earth.
Bacillus spiniferus (Unna).—
Rods sometimes curved, 2 p» in
length, -8 to 1 » in width, singly,
in pairs, and masses.
Colonies have peculiar spines, and
later a radiated marginal zone.
In the depth of gelatine minute,
yellowish, isolated colonies develop
in the track of the needle, and a
furrowed, yellowish-grey patch on
the free surface.
On agar the same yellowish
wrinkled growth appears.
On potato they form a shining,
faintly-yellow layer.
They were isolated from the skin
in eczema.
Bacillus spinosus (Luderitz)—
OF SPECIES,
Rods sometimes bent, 3 to 8 p» in
length, ‘6 » in width, and long
filaments. Spore-formation present.
They are anaerobic.
Colonies are composed of fine
radiating filaments, and liquefy the
jelly.
In agar the growth is composed
of colonies of matted filaments,
and there is gas-formation.
They liquefy blood serum.
They occur in earth.
Bacillus stolonatus (Adametz).
—Rods motile.
Colonies on gelatine, circular,
granular, and whitish, or yellowish-
brown. Colonies on agar send off
peculiar wavy processes.
Inoculated in the depth of gela-
tine a granular filament develops in
the track of the needle, and a white
patch on the free surface ; later the
gelatine is excavated in the upper
part, and the culture lines the
cavity.
On potato the growth is whitish.
They occur in water.
Bacillus stoloniferus (Pohl).—
Rods 1:2 » in length, °8 » in width.
Motile. :
Inoculated in the depth of gela-
tine they produce rapid liquefaction
in the track of the needle.
On the surface of agar they form
a white growth.
On potato they grow abundantly,
but scarcely at all in milk.
They occur in the water of
marshes.
Bacillus striatus albus (Bes-
ser).—Rods sometimes bent.
Colonies on gelatine appear as
minute dry points.
On agar the colonies have a brown
nucleus and clear marginal zone.
On the surface of agar the bacilli
produce a greyish-white thin layer.
On potato the growth is trans-
parent and slightly gelatinous.
They occur in nasal mucus.
Bacillus striatus flavus (Bes-
ser).—Short rods, straight or
curved ; involution forms.
Colonies granular, yellowish,
On the surface of agar they pro-
duce a white growth, which later
becomes sulphur yellow.
DESCRIPTION
On potato a similar colour is
produced.
They were isolated from nasal
mucus.
Bacillus subflavus (Zimmer-
mann).—Rods 1°5 to 3 w in length,
‘77 win width, and in chains. Motile.
Colonies prominent, yellowish-
white.
On the surface of gelatine they
form a yellowish-grey layer, and
on the surface of agar and potato
the growth is yellow.
They occur in water.
Bacillus subtilis (Hay bacillus).
—Cylindrical rods as much as 6 w
in length. Single forms grow to
double their length, and then
undergo division. They also form
threads which may be composed of
OESN
WS qe
ROUTE
Fic. 211.—Bacititus Susrinis witTH
Spores (BAUMGARTEN).
long rods, short rods, and cocci.
They are motile, and provided with
a flagellum at "each end. If the
nourishing medium is impoverished,
the multiplication of the rods by
division gradually ceases, and spore-
formation commences. The rods
become motionless, and a dark spot
is visible, either in the middle or
towards one end. This gradually
develops into a shining spore with
a dark outline. The rods swell
slightly during this process ; their
contour becomes undefined, and
soon disappears entirely ; spores
being set free in about twenty-
four hours. The spores are 1°2 p
long, and ‘6 » broad. They develop
into rods in the following way :—
On one side of the spore a swelling
appears, at the summit of which
an opening in the spore-membrane
results, and the germ escapes ; this
lengthens into a rod, and remains
for a time attached to the empty
spore-membrane.
OF SPECIES. 535
The spores are widely distributed,
and occur in the air, soil, dust, etc.
On the excrement of herbivorous
animals the bacilli form a white
efflorescence, and a thick crumpled
skin on liquid manure.
They flourish equally in liquids
and upon damp, solid, nourishing
media. They are aerobic ; depriva-
tion of oxygen causes the growth
of the bacilli to cease, and the rods
degenerate.
In plate-cultivations the colonies
are white, and, under a low power,
granular and irregular in outline
and faintly-greenish. Liquefaction
sets in, producing depressions like
‘saucers. The centre is opaque, and
is surrounded by a network of
filaments, which extend into the
gelatine surrounding the colony.
| iv
| i
i)
i
HA ! ll iI | /
ie ome )
Fic. 212.—PuRE-cULTURE OF BacrLLus
Sustitis in NuTRIENT GELATINE
(BAUMGARTEN).
Inoculated in the depth of gela-
tine, liquefaction occurs rapidly in
| the track of the needle, and a film
536 DESCRIPTION
floats on thesurface. The liquefied
gelatine, at first turbid, becomes
clear as the bacilli settle at the
bottom of the tube.
On agar a wrinkled film develops,
and also on serum.
Fic. 213. PuRE-cULTURE OF BaciLLus
SUBTILIS ON THE SuRFACE oF Nu-
TRIENT AGAR.
On potato the growth is white,
and there is copious spore-forma-
tion.
On ordinary nutrient liquids they
develop at first a thin, and subse-
quently a thick, dense, crumpled
pellicle, with copious spore-forma-
tion.
The simplest way to obtain a
culture of the bacillus is to make
a decoction of hay. The hay is |
chopped into small pieces, and
boiled with distilled water in a
flask for a quarter of an hour.
The infusion is then filtered into a
beaker, covered with a glass plate,
and set aside in a warm place. In
two or three days the liquid swarms
OF SPECIES.
with the bacilli, the spores of which
exist in great numbers in ordinary
hay. A more sure method for
obtaining a pure cultivation is as
follows :—
(a) Add only a small quantity of
water to some finely chopped hay,
and set aside for four hours at
36° C.
(b) Pour off the extract, and
dilute it to the sp. gr. 1-004.
(c) Boil gently for one hour in
a bulb plugged with cotton wool.
(d) Set aside 500 ccm. of the
extract at 36° C.
In about twenty-four hours, as
a rule, a pellicle has commenced
to develop upon the surface of the
liquid. If the reaction is definitely
acid, carbonate of soda solution
must be added to the decoction.
Metuops or Srarntina Hay Baciiuus.
To demonstrate the flagella of the
bacilli, they may be stained with
hematoxylin solution (Koch), or by
Loffler’s method.
The linking together of cocci, long
rods and short rods in the threads, is
shown by treating with alcoholic solu-
tion or fuchsine, or with iodine solution
(Zopf).
To stain the spores the cover-glass
preparations must be heated to a very
high temperature (210°C.), in the hot-
air steriliser for half an hour, or they
may be exposed for a few seconds to
the action of concentrated sulphuric
acid (Biichner), or floated for twenty
minutes on hot solution of the dye.
Bacillus subtilis similans—
| There are several bacilli closely
resembling Bacillus subtilis. /
Two have been isolated from
human feces by Bienstock which
do not liquefy nutrient gelatine.
No. I. Rods and filaments ;
spore-formation present.
On agar they produce a delicate
wrinkled veil.
No. II. Rods morphologically
identical with No. I.
On agar they produce a smooth,
shining layer.
Bacillus superficialis (Jordan).
—Rods 2°2 » in length, and ‘1 p in
width ; singly, and in pairs. Motile.
Colonies have a yellowish-brown
DESCRIPTION.
nucleus and transparent marginal
zone.
Inoculated in the depth of gela-
tine there is a slight growth in the
track of the needle, and after a time
liquefaction at the upper part.
On agar the growth is smooth and
shining.
In broth they produce turbidity.
They will not grow on potato.
They occur in sewage.
Bacillus tenuis sputigenus
(Pansini).—Short rods, singly, and
in pairs ; capsulated.
They produce a whitish growth
on the surface of gelatine.
They coagulate milk.
‘They are pathogenic in rabbits.
They were isolated from sputum.
Bacillus termo (Macé).—Thick
rods 1-4 » long, and -8 p wide,-
usually in pairs,sometimesin chains.
Actively motile.
Colonies whitish, with a grey edge
surrounded by liquefied gelatine.
Inoculated in the depth of gela-
tine they form a funnel-shaped
area of liquefaction, and later the
whole of the jelly is liquefied.
Broth is rendered turbid and a
thin brittle pellicle is formed. |
They are associated with decom-
position.
‘Bacillus tetani (p. 457).
Bacillus thalassophilus (Rus-
sel).—Slender rods varying in
length ; and filaments.
anaerobic. Spore-formation pre-
sent.
Wecutated in the depth of gela-
tine the growth appears in the
lower part of the track of the
needle“#a the form of cloudy colo-
nies, liquefying the jelly and pro-
ducing gas-bubbles. Cultures emit
a penetrating odour.
They were isolated from sea-mud.
Bacillus thermophilus (Mi-
quel).—Rods varying in size accord-
ing to the temperature at which
they are cultivated. In broth they
grow best between 65° and 70° C.,
forming a copious deposit. They
occur in air, soil, and water.
Bacillus tremelloides (Tils).—
Rods ‘75 to 1 » in length, °25 » in
width ; and in masses. ,
They are |
OF SPECIES. 537
Colonies
brown.
The bacilli inoculated in the:
depth of gelatine produce a growth
composed of isolated yellow colo-
nies in the track of the needle,
and a yellow mass on the surface.
They liquefy the gelatine.
On agar the growth is slimy and
golden-yellow. :
On potato they form an abund-
ant yellow growth.
They occur in water.
Bacillus tuberculosis (p. 378).
Bacillus tuberculosis galli-
narum (p. 402).
Bacillus tumescens (Zopf).—
Cocci, long and short rods. They
form a jelly-like disc °5 to 1 cm. in
diam. on slices of boiled carrot,
with the appearance of a rather
tough, crumpled skin of a whitish
colour. Examination of this pel-
licle shows that it is formed of
rows of rods lying closely together.
These rods can be observed to
divide into short rods and coeci.
Spore-formation occurs in two
stages of development—viz., in the
cocci and in the short rods. A
cultivation is obtained by exposing
slices of boiled carrot, slightly
moistened, to the air at the tem-
perature of the room.
Bacillus typhi
(p. 342).
Bacillus ubiquitus (Jordan). —
Rods 1:1 to 2m in length, -1 p»
in width ; and filaments.
Colonies granular
defined. j
The bacilli inoculated in the
depth of gelatine produce a growth
resembling that of Friedlander’s
pneumococcus. _
On agar and potato the growth
is greyish-white.
They coagulate milk and reduce
nitrates.
They occur in air and water.
Probably a variety of Bacillus
candicans. ;
Bacillus ulna (Cohn).—Cocci,
short rods, long rods, and threads.
Diam. of the cocci 1°56 to 2°2 yp.
Spore-formation in both short and
long rods. No septic odour is pro-
circular, _yellowish-
abdominalis
and well
538
duced by this bacillus in a nourish-
ing liquid. Cloudy masses are
found on the surface of the liquid,
which later form a thick dry
pellicle, consisting of bundles of
threads matted together. The for-
mation of ellipsoidal spores occurs
in the usual way; they measure
25 to 28 » long, and more than
1 » wide, The bacillus is found
in rotting eggs, and can be culti-
vated on boiled white of egg.
Bacillus ulna (Vignal) —Rods
2p in length ; singly, and in pairs,
and in short chains.
Colonies composed of concentric
zones varying in granularity.
Inoculated in the depth of gela-
tine, liquefaction occurs rapidly in
the track of the needle ; later, there
is a deposit at the bottom of the
liquefied area and a pellicle on the
surface.
On agar they form a white ad-
herent layer, and the jelly is tinged
with brown.
In broth a pellicle forms on the
surface.
On potato they form a pellicle
with characteristic linear markings.
They liquefy serum. Cultures
produce a putrefactive odour.
They occur in human saliva.
Bacillus vacuolosis (Sternberg).
—Rods 1°5 to 5 w in length, 1 » in
width, containing vacuolated proto-
plasm; filaments, and involution
forms. At times slowly motile.
Inoculated in the depth of gela-
tine, liquefaction occurs slowly at
the upper part of the track of
the needle, forming a cup-shaped
cavity ; the liquefied gelatine is
viscid, and a cream-white layer
forms on the surface.
In agar the development in the
track of the needle is scanty ; on
the surface a cream-white layer is
formed, and the bacilli are united
in long jointed filaments.
On potato a similar growth is
produced,
They were isolated from the in-
testine in fatal cases of ycllow fever.
Bacillus varicosus conjunctive
(Gombert).—Rods 2 to 8 » in length,
1 pin width.
DESCRIPTION OF SPECIES,
Inoculated in the depth of gela-
tine they produce a greyish-white
filament in the track of the needle,
and a greyish-white patch ‘on the
surface ; liquefaction follows with-
out turbidity.
On the surface of agar a white,
dry, adherent film is formed.
On potato the growth is, at first,
white and dry, later, reddish-brown.
They produce hyperemia when
injected into the conjunctiva. _
They were isolated from the
healthy human conjunctiva.
Bacillus venenosus (Vaughan).
—Motile rods.
Colonies circular, whitish.
Inoculated in the depth of gela-
tine there is growth in the track of
the needle and on the free surface.
' On agar they form a white film,
and on potato a moist brownish
layer.
They are pathogenic in small
animals.
They occur in water.
Bacillus venenosus brevis
(Vaughan).—Rods short and thick.
Colonies are yellow and composed
of concentric rings.
Inoculated in the depth of gela-
tine they grow in the track'of the
‘| needle and over the free surface.
On agar they produce a white
film.
On potato the growth is brownish.
They are pathogenic in small
animals,
They occur in water.
Bacillus venenosus invisibilis.
—Slender rods.
Colonies irregular, granular.
Inoculated in the depth of gela-
tine the growth is extremely slow
both in the track of the needle and
on the surface.
On agar there is a whitish film,
and on potatoes a brownish layer.
They are pathogenic in small
animals.
They occur in water.
Bacillus venenosus
faciens (Vaughan).-—Rods.
Colonies circular, granular, yel-
lowish.
Inoculated in the depth of gela-
tine they grow in the track of
lique-
DESCRIPTION OF SPECIES.
the needle and on the surface, and
liquefaction occurs after some
weeks.
On agar they produce a white
growth, and on potato it is brown-
ish or yellowish. i
They are pathogenic in small
animals.
They occur in water.
Bacillus ventriculi (Raczyn-
sky).—Rods 1:5 to 3 » in length,
1 » in width, singly, in pairs, and
in short chains.
Colonies have a dark nucleus and
transparent periphery.
On agar they forma white layer.
They were isolated from the
digestive tract of dogs.
Bacillus vermicularis (Frank-
land).—Large bacilli 2 to 3 » in
length, 1 p» in width, and long
threads. Spore-formation present.
Colonies are irregular in contour,
the irregularity increasing as the
colony comes to the surface. The
peripheral part is composed of
closely packed, wavy bands of bacilli,
and the centre is irregular and
wrinkled.
The bacilli inoculated in the
depth of gelatine form a flattened
band in the track of the needle, and
a grey layer on the surface ; lique-
faction slowly follows.
On agar they produce a smooth, .
shining, grey layer, and on potato
a thick, irregular, flesh-coloured
growth.
They reduce nitrates.
They occur in water. Probably
identical ‘with Bacillus vermicu-
losus.
Bacillus vermiculosus (Zimmer-
mann).—Rods 1°5 » in length, 85 p
in width, singly, in pairs, very short
chains and long filaments. They
are slowly motile.
: Colonies irregular ; grey, granu-
ar.
Inoculated in the depth of gela-
tine they produce, after four days,
liquefaction in the upper part of
the needle track, which spreads
downwards, and a reddish-grey sedi-
ment collects at the bottom of the
liquefied area.
On agar the growth is smooth and
539
| shining, and on potato yellowish-
grey.
They occur in water.
Bacillus violaceus (vide Bacil-
lus ianthinus).
Bacillus violaceus Laurentius
(Jordan).—Rods 3 to 3-6 » in length,
‘7 p in width.
Colonies violet, surrounded by
liquefied gelatine.
Inoculated in the depth of gela-
tine liquefaction occurs in the track
of the needle, and a violet sediment
collects at the bottom.
On agar the growth is violet, later
black.
On potato there is a copious
growth, changing in colour from
violet to black.
In broth a violet colour is pro-
duced in the presence of nitrates.
They coagulate milk, and render
it bluish-violet.
They occur in water. Probably
identical with Bacillus ianthinus
(Zopf). /
Bacillus virescens (Frick).—
Rods and filaments.
Colonies irregular,
een.
On the surface of gelatine they
colour the medium green.
They grow on agar.
On potato they form a brownish
growth.
In broth a pellicle is formed on
the surface, and beneath it the
liquid is coloured green.
They were isolated from green
sputum.
Bacillus viscosus (Frankland).
—Rods 1°5 to 2 yw in length, singly
and in pairs. F
Colonies granular, with hairlike
processes extending into the gela-
tine, which is liquefied and has a
green colour.
Inoculated in the depth of gela-
tine they produce liquefaction and
a green fluorescence.
On agar they form a greenish-
white layer, and colour the jelly
green.
On potato the growth is brown.
Probably identical with Bacillus
fluorescens liquefaciens.
Bacillus Zurnianus (List).—
granular,
540
Rods 1:2 to 1°5 win length, *6 to °8
in width. Colonies greyish-white,
viscid.
The bacilli inoculated in the
depth of gelatine develop slightly
in the track of the needle, and pro-
duce a prominent grape-like growth
on the free surface.
On potato the growth is grey or
tinged with yellow.
They occur in water.
Bacterium aerogenes (Miller).
—Short rods, singly and in pairs.
Motile.
The colonies are circular, well
defined, and yellowish.
Inoculated in the depth of gela-
tine the growth in the track of the
needle is brownish-yellow, and a
flat greyish button forms on the
free surface.
On agar a pulpy layer develops.
On potato the growth is pulpy
and yellowish-white.
The bacteria possess great power
of resisting the effect of acids.
They were isolated from the diges-
tive tract.
Bacterium brunneum
(Schréter).—Motile rods, produ-
cing a brown colour.
They were observed on a rotting
infusion of maize.
Bacterium decalvans (Thin).—
Cocci, singly or in pairs, 1°6 » in
length. é
They were observed in the roots
of the hair in cases of Alopecia
areata. ;
Bacterium fusiforme (Warm-
ing).—Rods_ spindle-shaped, with
pointed ends, 2°5 » long, and ‘5 to
“8 » thick. They were described
as forming a spongy layer on the
surface of sea-water.
Bacterium gingivae pyogenes
(Miller).-Shott rode, ee
The colonies are circular and
rapidly liquefy gelatine.
The bacteria inoculated in the
depth of gelatine produce rapid
liquefaction in the track of the
needle and a white sediment.
On agar they produce a moist
white growth.
They are pyogenic when -inocu-
lated subcutaneously in small ani-.
DESCRIPTION OF SPECIES.
mals, and cause a fatal result when
injected to the peritoneal cavity.
They occur in the deposit on the
teeth. ;
Bacterium hyacinthi (Wakker)..
— Cocci resembling Bacterium
termo.
They were observed in the yellow
slime of diseased hyacinth bulbs.
Bacterium —hydrosulfureum
ponticum (Zelinsky). — Long °
motile rods.
On agar a dark coffee-coloured
pigment is produced, which turns
black when exposed to air.
Cultures give off sulphuretted
hydrogen.
They were isolated from dredgings
in the Black Sea.
Bacterium litoreum (Warming).
—Cocci ellipsoidal, 2 to 6 » long,
1:2 to 2-4 w wide; singly, never
as chains or zooglea.
They occur in sea-water.
Bacterium luteum (List).—
Rods from 1:1 to 1°3 nlong. Non-
motile. The colonies are slimy, with
orange centres. :
Inoculated in the depth of
gelatine an orange growth occurs,
principally at the point of puncture.
Milk is coagulated.
They occur in water.
Bacterium merismopedioides
(Zopf).—Threads 1 to 15 uw in
thickness; these subdivide into
long rods, short rods, and finally
into cocci. The cocci divide first
in one and subsequently in two
directions, forming characteristic
groups, which appear like merismo-
pedia. These groups may eventually
consist of 64 by 64 cells or more,
and ultimately form zoogloea.. The
cocci develop again into rods and
threads.
They were observed in water
containing putrefying substances
(River Panke, Berlin).
Bacterium navicula (Reinke
and Berthold).—Cocci spindle-form
or ellipsoidal, including motile and
non-motile forms. They have one
or more dark spots, which may be
coloured blue by iodine.
They have been observed in rot-
ting potatoes.
DESCRIPTION
Bacterium photometricum (En-
gelmann).— Rods slightly reddish in
colour ; motile.
The movements are stated to
depend on light.
Bacterium synxanthum (Ehren-
berg ; Bacterium canthinum ; Bac-
terium of yellow milk).—Cocci °7 to
1 » in length, and rod-forms.
They produce a yellow colour in
boiled milk, which at first becomes
acid, and then strongly alkaline.
They also occur on boiled potatoes,
carrots, etc., where they form small
lemon-yellow masses.
The colouring-matter is soluble
in water, insoluble in ether and
alcohol, unchanged by alkalies, de-
colorised by acids. It is similar
to yellow aniline colours, both
spectroscopically and in ordinary
reactions.
Bacterium termo (Vignal).—
Rods 1:5 to 2 » in length, °5 to ‘7 p
in width.
Colonies white, surrounded by
liquefied gelatine.
The bacilli inoculated in the
depth of gelatine produce a funnel-
shaped area of liquefaction ; later,
the jelly is completely liquefied and
coloured green. Cultures have a
strong putrefactive odour.
In broth they form a white
deposit and colour the medium
green.
They were isolated from human
saliva.
Bacterium tholoeideum (Gess-
ner)—Rods similar to Bacillus
lactis aerogenes.
Pathogenic in small animals.
They were isolated from healthy
human evacuations.
Bacterium urex (Cohn).—Cocci
1:25 to 2 » in diam., singly or in
chains, and rods. The rods split
‘up by division into chains of cocci,
which after a time are set free. The
cocci increase further by subdivi-
sion, and a jelly-like membrane
develops around them. Masses of
cocci exist in the form of irregular
or roundish lumps. They are
aerobic.
Cultivations, after twenty-four -
hours, consist exclusively of rods ;
OF SPECIES, 541
after forty-eight hours, of cocci
chains; and in fourteen days, of
zoogloea ; the cocci transplanted
into fresh nourishing solution again
grow into rods. These observations
point to the existence of a pleo-
morphic species, Bacterium urece ;
and the former nomenclature, Jficro-
coccus ureee, taust be regarded as
untenable.
In urine they set up ammoniacal
fermentation, converting urea into
carbonate of ammonia. Rods, 2 »
long and 1 » wide, have been iso-
lated from stale urine (Bacillus
urez, Leube), which also most
energetically cause the ammoniacal
fermentation of urine.
Bacterium uree# (Jaksch).—
' Rods 2 y in length, 1 » in width.
Colonies on gelatine semi-trans-
parent.
Inoculated in the depth of gela-
tine the bacilli form a delicate
branching growth in the track of
the needle.
They convert urea into carbonate
of ammonia, and cultures smell of
herring brine.
They occur in ammoniacal urine.
Bacterium violaceum (Bergon-
zini)—Rods similar to Bacterium
termo, ‘6 to 1 » thick, 2 to 3 plong..
They occur on white of egg,
forming a violet pigment.
Bacterium Zopfii (Kurth).—
Cocci, 1 to 1:25 » in diam. ; rods
and threads. Cultivated in a streak
on nutrient gelatine spread out on
a glass slide, a peculiar develop-
ment takes place. In twenty-four
hours after inoculation threads
have developed; in forty-eight
hours windings of the threads
are observed, and in six days the
threads have broken up into cocci.
They were observed in the intestine
of fowls, especially in the contents
of the vermiform appendix. In-
oculation of rabbits was followed
by negative results. Identical with
Bacillus figurans (Crookshank).
Beggiatoa alba (Vauch).—Cocci,.
rods, spirals and threads (Fig. 215).
The threads are indistinctly articu-
lated, actively oscillating, and colour-
less; their protoplasm contains.
542
numerous strongly refractive gran-
ules consisting of sulphur. They
occur as greyish or chalk-white
gelatinous threads, 3 to 35 p
thick, in sulphur springs and
- moarshes.
Beggiatoa mirabilis (Cohn).—
Threads distinguished by their
breadth, which may reach 30 up.
They are motile, bent and curled
in various ways, and rounded at
the ends. Around the threads,
isolated cells have been observed,
DESCRIPTION OF SPECIES,
families, bound together by gela-
tinous substance. Later they be-
come larger, globular or ovoid in
shape, and hollow, containing
watery fluid in their interior. The
families reach a diameter of 660 p,
in which the cocci form simply a
peripheral layer. The hollow fami-
lies or vesicles are often perforated,
presenting a delicate reticulated ap-
pearance, which finally may become
broken up into irregular structures.
The red colouring-matter can be
Fic. 214.—Bactertum Zopru.
Successive CHANGES IN THE SAME THREAD,
- x 740. a, A thread form ; }, breaking up into rod forms; ¢, into cocci (Kurth).
macrococci, but spiral forms are as
yet unknown. The threads are
filled with sulphur granules. They
occur in sea-water, forming a white
gelatinous scum on decomposing
alge.
‘Beggiatoa roseo-persicina
(Cohnia roseo-persicina. Bacterium
rubescens, or Peach-coloured bac-
terium, Lankester).—Cocci, rods,
spirals, and threads (Fig. 216). The
cocci, globular or oval, reach 2°5 p
in diam. They form at first solid
distinguished from other red pig-
ments, and it is designated by the
name bacterio-purpurin. It is quite
distinct from the pigment produced
by Micrococcus prodigiosus, being
peach-blossom red, and_ insoluble
in water, alcohol, etc. Examined
spectroscopically, it shows a strong
absorption in the yellow, and a
weaker band in the green and blue,
as well as a darkening in the more
refrangible half of the spectrum.
In the cocci, especially of the older
DESCRIPTION OF SPECIES.
-vesicles, dark granules are to be
seen, which consist of sulphur.
‘The micro-organisms occur on the
surface of marshes, or on water in
which alge are rotting. They
form a rose-red, blood-red, violet-
red, or violet-brown scum; and
sometimes in such quantity that
543
Cladothrix dichotoma (Cohn).
—Threads resembling those of lep-
tothrix ; slender, colourless, not
articulated, ‘straight or slightly
undulated, and in places twisted
in irregular spirals with pseudo-
branchings. The development can
be traced from the cocci to rods and
Fic. 215.—BrcGIATOA ALBA.
‘A. Threads at base distinctly linked, partly spiral.
C, D. Fragments detached from threads ; immotile.
whole length.
B. A thread, spiral in its
E. Active
spirillum-forms, with a flagellum at either end. F,G. Thin and short spiral
forms.
whole marshes andi ponds may be
coloured blood-red by them.
Spirillum sanguineum, rosaceum,
violaceum, monas vinosa and Okeniil,
and Rhabdomonas rosea are pos-
sibly phase-forms of Beggiatoa
roseo-persicina.
H. A spiral showing the individual links.
x 540 (Zopf).
threads. The latter are at the
beginning simple threads, which
were formerly described as Lepto-
thrix parasitica, or, if coloured by
impregnation with iron, as Lepto-
thriz ochracea. Later they form
false branches by single rods turning
544
aside, which by repeated division '
lengthen into threads. A thread
appears to be first composed of
long rods, then of short rods, and
lastly of cocci. The iodine reaction :
must be applied to distinguish these
forms, especially when the sheath
of the threads has a yellow, rust-
red, olive-green, or dark brown
coloration. The cocci may grow
into rods while still in the sheath,
and finally become leptothrix
threads, surrounded by a delicate
gelatinous sheath, from which the
DESCRIPTION OF SPECIES,
media small tufts, about 1 to 3 p,
and floating masses.
Cladothrix Forsteri (vide Strep-
tothrix Férsteri, Cohn).
Cladothrix intricata (Russell).
—Rods and filaments.
Colonies are composed of a net-
work of twisted threads.
Inoculated in the depth of gela-
tine fine filaments spread out from
the track of the needle, and the
gelatine is liquefied.
Grown on agar the
penetrate the jelly.
filaments
Fic. 216.—PHASE-FORMS OF BEGGIATOA ROSEO-PERSICINA (WARMING).
false branching proceeds. Frag-
ments may break off, which are
actively motile, arid appear as
vibrios, spirilla, and spirocheta-
forms. They may also occur in
zoogloea (Fig. 217).
They are the commonest of all
bacteria in both still and running ©
water, in which organic substances
are present. They are observed
also in the waste water of certain
manufactures, such assugar. Arti-
ficially they can be cultivated on
infusions of rotting alge and ani-
mal substances, forming on these
In broth the growth is abundant.
They were isolated from sea
dredgings.
Cladothrix invulnerabilis (Ac-
osta, y Grande Rossi).—Filaments
‘ which produce in gelatine a white
. thread, and liquefy it very slowly.
On potato the growth is abundant
; and chalky in appearance.
In milk they form a firm yel-
lowish pellicle ; andin broth and in
water the growth is abundant.
They occur in water.
Clostridium butyricum (vide
Bacillus butyricus).
DESCRIPTION OF SPECIES. 545
Clostridium fetidum (Libo- gas-formation with unpleasant smell
rius).—Rods 1 » in width, singlyand and splitting up of the jelly.
in filaments. Spore-formation re- | They were isolated from earth.
sembles that of Bacillus butyricus. | Crenothrix Kihniana (Raben-
They are anaerobic. ' horst).—Cocci, rods, and thread-
Colonies rapidly liquefy gelatine. | forms. The cocci are globular,
te es
7 ete et OD
orm geeen een a&
ot
Fic. 217.—CLADOTHRIX DICHOTOMA.
A. Branching schizomycete :—(a) Vibrio-form ; (6) Spirillum-form [slightly mag-
nified].
B. A screw-form with («) Spirillum-form ; (b) Vibrio-form.
C. Long spirochzta-form. ate
D. Fragment with spirillum-form at-one end, vibrio-form at the other. .
# Screw-forms :—(a) continuous; (b) composed of rods ; (¢) composed of cocci.
Spirochzta-form :—(a) continuous ; (b) composed of long rods; (¢) short rods ;
(d) cocci (Zopf).
On agar the colonies form branch- | 1 to 6 »indiam. The threads are
ing processes resembling colonies of | colourless, 1-5 to 5 p thick, and club-
Bacillus cedematis maligni. | shaped at the extremity, reaching
Inoculated in the depth of gela- ; a diam. of 6 to9y. The threads
tine liquefaction spreads from be- | form colonies with a brick-red, olive-
low upwards. There is abundant . green, or dark-brown to brown-black
35
546 DESCRIPTION OF SPECIES.
coloration, caused by impreg- | set free when the sheath bursts,
nation with oxide of iron. The | and develop into new threads. In
threads are distinctly articulated, | other cases the segments remain
and ensheathed. The segments are | enclosed, and subdivide into discs,
@) Sef
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Th
Fic. 218.—CreNoTHRIxX KiMniana.
a, b, e, d, e. Cocci in various stages of fission, x 600.
f. Googloea of cocci, x 600.
g. Various forms of zooglea, natural size.
h. Colony of threads composed of rods grown out of a zooglcea of cocci.
i—r. Thread-forms ; some straight, others spiral, with more or less differentia-
tion between base and apex. (7) is composed of short rods at the base, and above
these of cylindrical segments, and at the apex these segments have divided into
cocci, x 600 (Zopf).
&
DESCRIPTION
which, by vertical fission, break up
into globular forms (cocci). These
again develop into new threads,
either within the sheath, eventually
penetrating it, or after they are set
free.
The micro-organism appears in
little whitish or brownish tufts in
wells and drain-pipes, and it not
only renders drinking-water foul,
but may stop up ‘the narrower
‘Pipes
iplococcus albicans amplus
(Bumm).—Cocci resembling gono-
cocci but much larger, singly and
in tetrads.
Colonies are prominen
greyish-white.
In the depth of gelatine they
produce a greyish-white growth in
the track of the needle and on the
free surface. They slowly liquefy
the gelatine.
They were obtained from the
vaginal mucous membrane.
Diplococcus albicans tardis-
simus (Bumm).—Cocci morpho-
logically identical with gonococci.
They grow extraordinarily slowly
on gelatine.
They form very minute colonies,
which are opaque and greenish-
brown in colour.
Inoculated in the depth of gela-
tine isolated greyish-white colonies
develop in the track of the needle,
and on the free surface a thin,
white, waxy film with dentated
edge.
On agar the growth is very
similar.
They were isolated from the
vaginal mucous membrane.
Diplococcus citreus conglome-
ratus (Bumm).—Cocci in pairs '
resembling gonococci, 1°5 « in diam.,
in tetrads and in masses.
Colonies lemon-yellow ; irregular
in form.
Inoculated in the depth of gela-
tine the cocci develop in the track
of the needle, and liquefaction com-
mences at its upper part.
The growth on the free surface
is yellow, and floats on the liquefied
gelatine or subsides to the bottom
of the liquefied area.
and
OF SPECIES. 547
They are present in gonorrhceal
pus, in air and in dust.
Diplococcus citreus lique-
faciens (Unna)—Oval cocci -4 to
‘1 » in diam., in pairs, tetrads,
short chains, and masses.
Colonies appear in the form of
circular discs, at first greyish-white,
later lemon-yellow. They are finely
granular, and have sharply defined
contours.
Inoculated in the depth of gela-
tine, at the end of a week the
growth is found on the free
surface, forming a shining yellow
layer; in two weeks liquefaction
commences, and the growth floats
on the liquefied gelatine which is
also yellowish and turbid.
On the surface of agar a yellow-
ish-brown layer is rapidly formed.
The appearance is similar on
potato.
They were isolated in cases of
eczema seborrheicum.
Diplococcus coryze (Hajek) —
Large diplococci.
Colonies are white, prominent.
Inoculated in the depth of gela-
tine the growth resembles the
pneumococcus.
On agar a white layer is formed.
They are probably identical with
Friedlinder’s pneumococci.
They were isolated from the
mucus in acute nasal catarrh.
Diplococcus flavus liquefaciens
*tardus.—Cocci resembling gono-
cocci.
Colonies are circular, shining,
and chrome-yellow in colour.
Inoculated in the depth of gela-
tine a yellowish growth occurs
along the needle track, and also
on the free surface. In a month
the surface is depressed, but the
gelatine is not liquefied until about
two months have elapsed.
On agar a yellowish-white layer
is formed, and on potato the colour
is more proneunced.
They were isolated from the
skin in eczema seborrhoeicum.
Diplococcus fluorescens feti-
dus (Klamann).—Cocci in pairs and
chains. ;
Colonies circular, forming a
548 DESCRIPTION
brownish deposit surrounded by
liquefied gelatine which has a violet
or greenish tinge.
Inoculated in the depth of gela-
tine they produce liquefaction along
the track of the needle, with a
hemispherical excavation of the
gelatine at the upper part. An
iridescent film floats on the surface
and a greenish sediment forms at
the bottom of the liquefied area.
On agar the layer is brownish,
On potato the growth is granular,
and the potato in the vicinity has
a bluish colour.
They were cultivated from the
nasal mucus.
Diplococcus intercellularis ©
meningitidis (Weichselbaum).—
Cocci singly, in pairs, tetrads and
masses. They grow at 37°C.
Colonies on agar are granular and
yellowish-brown.
On the surface of agar they form
a greyish-white viscid growth. In
the depth of agar the growth only
occurs in the upper part of the
needle track.
On blood serum and broth there
is very little growth, and none on
potato.
Cultures quickly lose their
vitality.
They are pathogenic in mice,
guinea-pigs, rabbits, and dogs.
They were isolated from the exu-
dation in cases of cerebro-spinal
meningitis, and were observed in
the interior of pus cells.
Diplococcus luteus (Adametz).
—Cocci 1-2 to 1'3 » in diam., singly
and inchains. Motile.
Colonies are circular and slightly
yellow, and granular. Old colonies
are bright yellow.
On the surface of gelatine a
growth occurs in concentric circles
of a lemon-yellow colour, and the
gelatine is coloured reddish-brown.
After several weeks liquefaction
sets in. :
On agar a yellow layer forms, and
the jelly is coloured reddish-brown.
On potato the growth changes
from yellow to brown. Milk is
coagulated.
They were obtained from water.
OF SPECIES.
Diplococcus of pneumonia in
horses (Schutz).—Oval cocci, singly
or in pairs, capsulated.
Colonies small and white.
In the depth of gelatine a row of
colonies develops in the track of the
needle,
On agar the growth is composed
of transparent droplets.
Injection into the lung is said to
produce pneumonia, ending fatally
in eight or nine days.
They are pathogenic in rabbits,
guinea-pigs, and mice.
They were isolated from the
lungs of a horse suffering from
‘pneumonia,
Diplococcus roseus (Bumm).—
Cocci identical in description with
gonococci.
Colonies are pink, granular, and
irregular in form.
Inoculated in the depth of gela-
tine the cocci grow freely in the
track of the needle and on the
surface, developing a pink colour
and slowly producing liquefaction.
They are present in the air.
Diplococcus subflavus (Bumm).
—Diplococci similar to gonococci.
Colonies greyish-white, later
yellow.
They grow in gelatine and on
blood serum, and liquefy broth.
They produce suppuration when
injected subcutaneously in man.
They were isolated from lochial
discharges, the vesicles of pem-
phigus, and from the secretion in
colpitis in children.
They stain by Gram’s method.
Hematococcus bovis (Babés).—
Cocci oval, singly, in pairs, and in
masses.
Inoculated in the depth of gela-
tine minute colonies develop in the
track of the needle.
On agar the growth is composed
of transparent droplets.
On potato a yellowish shining
film is formed.
On blood serum the growth is
similar to that on agar.
They produce a fatal result in
rabbits and guinea-pigs in a week
or ten days.
They were isolated from the
DESCRIPTION
blood and organs of cattle which
died of an epidemic disease asso-
ciated with hemoglobinuria.
Helicobacterium aerogenes
(Miller).—Bacilli singly, in chains
and long wavy filaments. Motile.
Colonies whitish, varying in form.
Inoculated in the depth of gela-
tine the bacilli give rise to a faintly
yellow growth in the track of the
needle, and an almost invisible,
rapidly growing layer on _ the
surface.
On potato the growth is dry and
brownish.
They were isolated from the
healthy intestinal tract.
Leptothrix buccalis (Robin).—
Long, thin threads, -7 to 1 » broad,
colourless, often united in thick
bundles or felted together. Masses
of cocci occur with the threads,
and the threads themselves are com-
posed of long rods, short rods, and
cocci, The threads may break up
into spiral, vibrio, and spirochata
forms. The last-named occur in
large numbers in the mouth, and
have been named Spirocheta buc-
calis. Weptothrix buccalis is found
in teeth slime, and is believed
to be intimately connected with
dental caries. The threads pene-
trate the tissue of the teeth, after
the enamel has been acted upon by
acids generated by the fermentation
of food. The short rods, long rods,
cocci, leptothrix-forms, and screw-
forms are found in the dental
canals.
The threads of Leptothrix buc-
calis have a special staining reaction
(Leber). They become coloured if
placed in an acid medium with
iodine ; if the medium be alkaline,
it must first be acidified with very
dilute hydrochloric acid or acetic
acid. The contents are stained
violet, and contrast with the sheath
and septa, which remain uncoloured.
Leptothrix buccalis (Vignal).—
Rods 1 to 1°5 » in width, 1°6 to
30 pw in length.
Colonies greyish-white, promi-
nent and furrowed.
Inoculated in the depth of gela-
tine a filament forms in the track
OF SPECIES. 549
of the needle, and a growth occurs
on the free surface. Liquefaction
sets in at the upper part, forming
a cup-shaped cavity, and a bluish
skin floats upon the liquid. The
liquefaction gradually extends to
the side of the tube, and a deposit
is formed at the bottom of the
liquefied gelatine.
On agar the layer is white,
wrinkled and transparent, and later
yellowish.
In broth there is turbidity, but
no skin on the surface.
On potato the growth is greyish-
white.
They are occasionally present
in the mouth in health, and
are possibly identical with lepto-
thrix buccalis (Robin).
Leptothrix gigantea (Miller).—
Long rods, short rods and cocci
can be observed in the same thread.
There are also screw-threads, which
may take the form of spirals,
vibrios, or spirochete. The threads
increase in diameter from base
to apex ; corresponding with the
thickness of the threads, the rods
and cocci show different dimensions.
They have been observed in the
diseased teeth of dogs, sheep, cats
and other animals.
Leuconostoc mesenteroides,
Cienkowski (Gomme de sucrerw,
Froschlaichpilz, Frogspawn fungus).
—Cocci and rods singly, in chains,
and in zoogloea, surrounded by a
thick gelatinous envelope. The life-
history has been very thoroughly
investigated. The spores, 18 to
2 p in diam., are of a round or
ellipsoidal form, with thick mem-
brane and shining contents. The
outer membrane-layer bursts, and a
middle lamella oozes out, and forms
a thick gelatinous envelope, while
the inner layer remains adherent
to the plasma. Thus the spore-
germination leads to the formation
of a coccus with a gelatinous en-
velope. The coccus then elongates
into a short rod-form, and the
gelatinous envelope becomes ellip-
soidal. The rod divides into two
cocci, and each of these lengthens
intoarodand divides. By repetition
550 DESCRIPTION
of this process a chain of cocci
results, encased in a cylindrical
or ellipsoidal envelope. The chains
increase in length, become twisted
up, and eventually fall apart into
pieces of various lengths.
In nourishing liquids a great
number of little masses are formed,
which adhere together, and produce
pseudo-parenchymatous structures.
These latter may join together,
forming still larger agglomerations.
+
2° ©
OF SPECIES.
This micro-organism occurs occa-
sionally in beet-root juice and the
molasses of sugar-makers, forming
large gelatinous masses resembling
frog-spawn. The vegetation is so
rapid that forty-nine hectolitres of
molasses, containing 10 per cent.
of sugar, were converted within
twelve hours into a gelatinous
mass ; consequently, it is a for-
midable enemy of the sugar manu-
facturers.
Fic. 219.—LEvcoNnostoc MESENTEROIDES.
1. Spores. 2. Spores after germination, showing gelatinous envelope. 3, 4, 5, 6.
Increase by division.
mass of zoogloa.
kowski).
The masses of zoogloea are of
almost a cartilaginous consistency,
and admit of sections being made
with a razor. After a long time
the envelope liquefies, and the cocci
are set free ; the latter introduced
into fresh nourishing media develop
new colonies, In the chains some
of the cocci become enlarged with-
out changing their' form. These
acquire the properties of spores,
and are arthrospores.
7. Glomerular form of zoogloa.
9. Cocci chains with arthrospores (Tieghem and Cien-
8. Section of an old
Micrococcus acidi lactici
(Marpmann).—Large cocci, singly
and in pairs.
Colonies yellowish-white.
On the surface of gelatine the
cocci produce a yellow layer.
They grow in milk, producing a
reddish colour, and coagulation due
to the formation of lactic acid.
They were isolated from milk.
Micrococcus acidi lactici lique-
faciens (Kreuger).—Cocci oval, 1
DESCRIPTION OF SPECIES.
to 1-5 » in diam,, in pairs and in
tetrads.
Colonies white.
Inoculated in the depth of gela-
tine the cocci produce a granular
filament, changing in two or three
days to liquefaction in the form of
a funnel; later, a wrinkled mem-
brane floats on the surface of the
liquid.
In milk they produce lactic acid.
They were isolated from butter
which had turned cheesy.
Micrococcus aerogenes (Miller).
—Oval cocci.
Colonies are dark and regular in
contour, but have a peculiar spotted
appearance.
Inoculated in the depth of gela-
tine «a brownish-yellow growth
occurs along the track of the needle,
and on the free surface a white
button-like elevation. After a time
the gelatine is slowly liquefied.
On agar a yellowish-white pulpy
layer forms, and a similar growth
appears on potato.
They resist the action of acids, so
that the presence of gastric juice
does not impede their develop-
ment.
They were obtained from the
intestine.
Micrococcus agilis (Ali-Cohen).
—Cocci 1 » in diam., singly, in
pairs, tetrads, and in chains. They
are motile, and possess flagella.
Inoculated in the depth of gela-
tine they grow in the track of the
needle, and produce, after two or
three weeks, liquefaction or excava-
tion of the jelly. .
On agar and potato the growth
is pink.
They occur in water.
Micrococcus agilis citreus
(Menge).—Cocci in pairs, chains
and masses. They are motile, and
each coccus possesses a single fla-
gellum.
Colonies appear surrounded by
clouded gelatine.
Tnoculated in the depth of gela-
tine there is a scanty growth in the
track of the needle, and on the sur-
face a bright yellow patch.
On agar they form a yellow layer,
551
which is viscid, and may be drawn
out in long threads.
In broth they produce cloudiness
and a viscous deposit.
The growth on potato is bright
yellow.
Milk is not coagulated.
They were isolated from an in-
fusion of peas.
Micrococcus albus liquefaciens
(Besser ).—Large cocci in chains and
in masses. They are anaerobic.
Colonies on agar exhibit concen-
tric rings of different shades of
brown.
Inoculated in the depth of gela-
tine they produce liquefaction in
the track ot the needle.
They occur in mucus from the
nose.
Micrococcus amylivorus (Bur-
rill).—Oval cocci, 1 to 1:4 » long,
‘7 » broad, singly, in pairs, and rarely
in fours, never in chains, are found
embedded in an abundant mucilage
which is very soluble in water.
They have been described as pro-
ducing the so-called ‘fire blight”
of the pear tree and other plants.
Micrococcus aquatilis (Bolton).
—Small cocci in masses.
Colonies circular, prominent, and
pure-white.
Inoculated in the depth of gela-
tine, there is a white growth in the
track of the needle and also on the
free surface.
On agar the growth is white.
They occur in water.
Micrococcus aquatilis invisi-
bilis (Vaughan).—Cocci oval.
Colonies brown.
In gelatine there is a slight
growth in the track of the needle,
and a more abundant growth on
the free surface.
On agar they form a white film.
On potato the growth is in-
visible.
They occur in water.
Micrococcus aurantiacus
(Cohn).—Cocci spherical or oval,
1-3to 1:5 win diam., singly, in pairs,
and in groups.
Colonies orange-yellow.
Inoculated in gelatine they form
minute colonies in the track of the
552 DESCRIPTION
needle, and a prominent hemi-
spherical yellow growth on the free
surface.
On agar the growth is orange-
yellow, and on potato yellow and
slimy.
They occur in water.
Micrococcus botryogenus
(Johne, Rabe).—Cocci 1 to 1°5 p in
diam., in wavy chains.
Colonies circular, sharply defined.
At first silver-grey, later yellowish-
grey with metallic lustre, they
produce an odour like that of
strawberries.
Inoculated in the depth of gela-
tine a greyish-white filament de-
velops, with slight liquefaction of
the gelatine; later, it becomes
milk-white, and at its upper part a
characteristic bubble appears.
They make hardly any growth
on agar.
On potato they grow very abun-
dantly, forming a yellowish layer
with the same odour as the colonies
on plate cultivations.
Inoculated guinea-pigs die of
septicemia ; in sheep and goats
severe inflammation spreads from
the point of inoculation. Mice are
immune. In horses an inflamma-
tory cedema is at first set up,
followed in four to six weeks by
the formation of new growths,
which sometimes suppurate and
contain large numbers of micro-
cocci.
They were found in tumours of
the spermatic cord and of the
connective tissue in other parts in
horses.
Micrococcus candicans
(Fliigge)—Cocci which collect in
masses.
In plate-cultivations they form
in two or three days milk-white
colonies; while those below the
surface of the gelatine are yellow-
ish. Under a low power the deep
colonies are quite circular, with
smooth margins, of a blackish-brown
colour, and very slightly granular ;
the superficial colonies are quite
irregular in outline, and are finely
granular.
Cultivated in test-tubes they form
OF SPECIES.
a white nail-shaped cultivation.
They were isolated from contami-
nated plate-cultivations.
They occur in the air.
Micrococcus candidus(Cohn).—
Cocci forming snow-white points
and spots upon slices of cooked
potato.
Micrococcus carneus (Zimmer-
mann).—Cocci °8 » in diam., occur-
ring in masses.
Colonies circular, greyish-white,
with the centre tinged with red.
Inoculated in the depth of gela-
tine they form a white, granular
filament in the track of the needle,
and a pale pink layer on the free
surface.
On the surface of oblique gela-
tine a flesh-coloured layer develops,
which later assumes a violet colour.
On agar the growth is similar.
On potato the growth is abundant
and red in colour.
They were isolated from water.
Micrococcus cerasinus siccus
(List).—Cocci -25 to *12 » in diam.,
singly and in pairs. They can best
be cultivated at 37° C.
On agar they form a cherry-red
layer, and a similar growth on
potato.
The pigment is insoluble in alco-
hol, ether, and water, and is not
destroyed by acids or alkalies.
They occur in water.
Micrococcus cereus albus
(p. 178).
Micrococcus cereus flavus
(p. 178).
Micrococcus cinnabareus
(Fliigge).—Large cocci occurring in
twos, threes, and fours.
Colonies develop very slowly, and
are punctiform, and bright red at
first, and afterwards reddish-brown.
The cocci inoculated on the sur-
face of gelatine form a heaped-up,
red-coloured growth. ‘
They were found contaminating
old cultivations.
Micrococcus citreus (List).—
Cocci 1°5 to 2:2 » in diam., singly,
in pairs and chains.
Colonies are irregular in form,
moist and shining, and yellowish in
colour,
DESCRIPTION OF SPECIES.
In the depth of gelatine the
growth is very scanty.
On the surface of agar the growth
is yellowish.
On potato the growth is similar
but more abundant.
Micrococcus concentricus
(Zimmermann).—Cocci ‘9 pin diam.,
in masses,
Colonies bluish-grey.
Inoculated in the depth of gela-
tine there is no growth in the track
of the needle, but concentric rings
form on the free surface.
On agar the growth is greyish-
white and smooth.
On potato yellowish and slimy.
They occur in water.
Micrococcus cremoides (Zim-
mermann).—Cocci ‘8 » in diam.,
occurring in masses.
Colonies are spherical, granular
and yellowish. The margins are
dentated or irregular, and processes
extend intothe surrounding gelatine.
Inoculated in the depth of gela-
tine liquefaction occurs in the
track of the needle in a few days.
A yellow growth floats on the
liquefied gelatine, and a yellowish
mass subsides to the bottom of the
liquid.
On agar a smooth shining layer
is formed, and on potato the growth
is abundant.
They occur in water.
Micrococcus crepusculum
(Cohn. Monas crepusculum, Ehren-
berg. Mikrokokken in faulenden
Substraten, Fligge).—Round or
short oval cells, scarcely 2 » in
diam. ; singly or in zooglea.
They occur in various infusions
and putrefying fluids in company
with Bacterium termo.
Micrococcus cumulatus tenuis
(Besser).—Large cocci, oval; in
masses.
Colonies on agar have a brown
nucleus.
Inoculated in the depth of gela-
tine they form a white filament,
and on the surface a transparent
layer.
In broth there is an abundant
deposit, and the supernatant liquid
is clear.
053 +
They occur in mucus from the
nose.
Micrococcus endocarditidis ru-
gatus (Weichselbaum).—Cocci re-
sembling pyogenic staphylococci.
Colonies have a brown or yellow-
ish-brown nucleus.
In the depth of agar there is a
slight growth in the track of the
needle, and a wrinkled, waxy layer
on the surface,
On potato the growth is dry and
brownish.
On blood serum the growth is
colourless and adherent.
Injected subcutaneously, they pro-
duce, in rabbits, local swelling and
redness, and suppuration in guinea-
pigs. Injected into the veins after
injury to the aortic valves, they
produce endocarditis.
They were isolated from a case
of ulcerative endocarditis.
Micrococcus fervidosus (Ada-
metz).—Cocci ‘6 » in diam., in pairs
and in masses.
The deep colonies are pale-yellow,
and look like watery droplets ; but
superficial colonies are granular and
irregular with jagged edges.
Inoculated in the depth of gela-
tine a granular filament develops in
the track of the needle, and on the
surface a circular patch with den-
tated margin.
On agar the growth is white and
slimy, and on potato greyish-
white.
They occur in water.
Micrococcus Finlayensis
(Sternberg).—Cocci 5 to ‘7 p in
diam., singly, in pairs, tetrads, and
in masses.
In the depth of gelatine they
produce a growth in the track of
the needle, with liquefaction at
the upper part with a pale-yellow
deposit.
On agar the growth is pale-yellow.
They were isolated from the liver
in a fatal case of yellow fever.
Micrococcus flavus desidens
(Fligge).—Cocci singly, in- pairs,
or chains of a few elements.
Colonies yellowish-white.
Incculated in the depth of gela-
tine they grow along the track of
554
the needle, and form a yellowish-
brown layer at the point of punc-
ture.
Later liquefaction sets in, and a
deposit forms at the bottom of the
turbid liquid.
They occur in air and in water.
Micrococcus flavus lique-
faciens (Fliigge).—Cocci mostly in
twos and threes, also in masses.
Small yellow colonies appear after
two or three days, which have a
shallow depressed zone surrounding
them. Under a low power they
are granular and yellowish-brown,
with lines radiating from the centre |
to the circumference. Later they
liquefy the gelatine, and coalesce,
Inoculated in the depth of gelatine
the cocci produce spherical yellow
colonies in two days along the track
of the needle. These become con-
fluent, and at the end of eight days
the whole of the jelly has become
liquid ; later the upper part becomes.
clear, and a yellow mass subsides to
the bottom of the tube.
They occur in air and in water..
Micrococcus flavus tardigra-
dus (Fliigge).— Large cocci showing |.
at times peculiar dark poles ; gener-
ally arranged in masses.
Colonies develop slowly; the
superficial ones have a smooth wax-
like surface with projecting centre ;
those below the surface are of a
dark chrome-yellow colour, and are
round or oval.
Inoculated in gelatine the cocci
develop slowly along the track of
the needle, forming small isolated
colonies; the gelatine is not
liquefied.
They occur in air and in water.
~~ Micrococcus fetidus(Klamann).
—Cocci singly, in pairs, and short
chains and masses.
Colonies circular or oval, white.
Inoculated in the depth of gela-
tine a pure white, shining growth
forms in concentric circles at the
point of puncture, and develops a
brownish colour ; and liquefaction
occurs after a time, and extends
along the needle track.
A white layer spreads over the
surface of agar.
DESCRIPTION
OF SPECIES.
On potato the growth is slimy
and grey in colour, with a red tinge.
Cultures produce an odour like
that of ozzna.
They were isolated from the
nose.
Micrococcus fcoetidus (Rosen-
bach).—Small oval cocci.
Cultivated in agar-agar they
develop gas-bubbles and a foetid
odour. They were isolated from
carious teeth.
Micrococcus Freudenreichi
| (Guillebeau).—Large cocci, singly
and, in chains.
Colonies are granular and puncti-
form.
In broth turbidity is produced,
and, later, a flocculent deposit.
On potato a shining film develops,
yellowish or brownish-yellow in
colour.
In milk the cultures become
viscous, and can be drawn out into
threads several yards in length.
They were isolated from milk
with viscous fermentation.
Micrococcus fuscus (Maschek).
—Cocci oval.
Colonies pale-brown or black.
Inoculated in the depth of gela-
tine there is a slight growth along
the track of the needle, and a brown
layer forms on the surface followed
by liquefaction.
On potato the growth is brown
or brownish-black and slimy.
Cultures give off an odour of
putrefaction.
They occur in water.
Micrococcus gingive pyogenes
(Miller).—Large cocci, singly and
in pairs.
Colonies spherical, with sharp
contours. ©
Inoculated in the depth of- gela-
tine there is an abundant growth
along the track of the needle and
on the free surface.
On agar a thick film develops,
with a faint tinge of purple by
transmitted light.
Injected into mice subcutaneously
they produce local suppuration, and
sometimes death. Injected into
the peritoneal cavity they produce
_ peritonitis and death.
DESCRIPTION OF SPECIES.
They were isolated from an
abscess of the gums.
Micrococcus gonorrhee
(p. 190).
Micrococcus havaniensis
(Sternberg).—Cocei 4°5 » in diam.
The colonies are circular and of
a blood-red colour,
The cocci inoculated in the depth
of gelatine produce a colourless
growth in the track of the needle
and a carmine patch on the surface.
On agar and on potato they form
a thick irregular carmine layer.
Micrococcus in Biskra-button
(Heydenreich).—Cocci in pairs,
*86 to 1 » in length, occasionally
tetrads ; capsulated.
Inoculated in the depth of gela-
tine they form a greyish-white fila-
ment composed of closely packed
colonies, and a yellowish-white film
on the free surface. Liquefaction
commences at the upper part of the
needle track in a few days, forming
a funnel which extends until, in
two weeks, the gelatine is com-
pletely liquefied.
On the surface of agar a shining
white or yellowish-white layer de-
velops in twenty-four hours.
On potato the growth is similar.
Inoculations are said to produce
in rabbits, dogs, fowls, sheep and
horses a morbid condition of the
skin similar to the disease known as
Biskra-button or Pendjeh sore. In
man they produce suppuration when
rubbed on the skin.
They were isolated from the
disease known as Pendjeh sore,
Biskra-button or alow de Biskra.
Micrococcus in gangrenous
mastitis in sheep.—Cocci singly,
in pairs, and in masses.
Colonies are spherical, white, and
under a low power have a brown
nucleus and transparent margin.
The cocci inoculated in the depth
of gelatine produce a conical area
of liquefied jelly with a copious
white deposit.
On agar they produce a white
layer, which later turns yellowish
in colour.
On potato they form a greyish
growth.
555
Injected into the mamary gland
of sheep they produce inflammatory
cedema, and a fatal result in
twenty-four to forty-eight hours.
In rabbits they are pyogenic.
They were isolated from the milk
in cases of gangrenous mastitis in
sheep.
Micrococcus in infectious
pleuro-pneumonia (Poels and
Nolen)—p. 242.
Micrococcus in influenza (Fis-
chel).—-Cocci from 1 to 1:25 p» in
diam., singly, in pairs, and chains.
Extremely minute colonies appear
in three days.
Inoculated in the depth of gela-
tine a milk-white filament forms
along the track of the needle. Lique-
faction commences in four days
at the upper part, and extends
slowly.
On agar the colonies are pure-
white.
On potato the growth is yellowish-
white.
They do not grow on blood serum
or in milk. \
Intravenous injection in dogs is
said to produce symptoms like
distemper.
They were obtained from the
blood in cases of influenza.
Micrococcus in influenza
(Kirchner).—Cocci in pairs and
chains; capsulated. They grow at
387° C.
The colonies are transparent,
whitish.
On the surface of agar there is
an abundant growth, but it is
limited in the depth of the jelly.
Inoculation experiments were
inconclusive.
They were obtained from the
sputum in cases of influenza. —
Micrococcus in pemphigus
(Almquist).—Cocci ‘5 to 1 pm in
diam., singly and in pairs ; identi-
cal with Staphylococcus pyogenes
aureus.
The cocci vaccinated on the arm
are said to have produced bulle.
They were isolated from pem-
phigoid bulla in children. :
Micrococcus in pemphigus
(Demme).—Cocci ‘8 to 14 » in
556 DESCRIPTION
diam., singly, in pairs, and in
masses. They can be cultivated
at 37° C.
The colonies on agar are milk-
white and prominent. Later, off-
shoots occur from the margin,
producing a rosetted appearance.
Inoculated in the depth of
gelatine the cocci form clubbed or
stalactitic out-growths from the
filament which develops in the
track of the needle.
On the surface of agar a creamy
layer is formed with similar off-
shoots.
Injected into the lungs of guinea-
pigs they are said to produce
broncho-pneumonia.
They were obtained from the
bull in acute pemphigus.
Micrococcus in pneumonia
(Manfredi).—Oval cocci ‘6 to 1
in width, 1 to 1°5 p in length, singly,
in pairs, and short chains.
Colonies on gelatine are circular,
whitish, and later spread out and
become bluish by transmitted light,
and of a pearly lustre by reflected
light.
Inoculated in the depth of gela-
tine there is a limited growth along
the track of the needle.
On blood serum they form a
shining, granular, faintly greenish-
yellow layer.
They also can be cultivated on
potato and in broth.
They are pathogenic in dogs,
rabbits, guinea-pigs, mice and birds.
Birds die in a few days; mammals
in from one to three weeks. After
death new growths composed of
granulation tissue are found in the
internal organs, varying in size
from a millet seed toa pea. They
were obtained from the sputum of
pneumonia complicating measles.
Micrococcus in progressive
abscess formation in rabbits
(Koch).—Coceci only about +15 » in
diam., principally in thick zoogloea.
The disease was induced by the in-
jection into rabbits of decomposing
blood. At the place of injection a
spreading abscess formed, which was
fatal to the animal in about twelve
days. No bacteria were observed
OF SPECIES.
in the blood, but in the walls of the
abscess thick masses of cocci were
found. The pus is_ infectious,
causing the same disease in healthy
rabbits.
Micrococcus in pyemia in
rabbits (Koch).—Round cocci and
diplococci ‘25 p in diam.
The disease was produced by the
subcutaneous injection, in a rabbit,
of distilled water in which the skin
of a mouse had been macerated.
At the autopsy there were found
great infiltration around the site of
injection, peritonitis, and accumu-
lations in the liver and lungs; in
short, the appearances of pyzmia.
Fic. 220.—Micrococcus in Pyzmra
In RasBits: VESSEL FROM THE
Cortex or THE KipNEY x 700.
a, Nuclei of the vascular wall;
c, Masses of micrococci adherent
to the wall and enclosing blood-
corpuscles (Koch). y
In the capillaries of the organs
examined, masses of cocci were
observed enclosing blood-corpuscles.
Fresh inoculations in rabbits with
exudation-fluid, or blood from the
heart, reproduced the same disease.
Micrococcus in septicemia in
rabbits.—Ellipsoidal cocci 8 to 1
p in largest diam. The disease
was produced by the injection of
putrid meat infusion. After death
slight cedema was noted at the site
of injection, slight extravasation of
blood, and great enlargement of
the spleen. No emboli or peri-
tonitis resulted. Masses of cocci
were found in the capillaries of
DESCRIPTION OF SPECIES.
different organs, especially in the
glomeruli of the kidneys. Rabbits
and mice inoculated with blood
from the heart proved susceptible
_to the disease.
Micrococcus in syphilis (Disse
and Taguchi).—Cocci and diplo-
cocci.
They produce a greyish-white
growth in nutrient media. |
They are said to produce inflam-
matory changes in the internal
organs and disease of the blood-
vessels when inoculated in dogs,
rabbits and sheep.
They were obtained from the
blood in cases of syphilis.
Micrococcus in trachoma
(p. 190).
Micrococcus in yellow fever
(p. 260).
Micrococcus lactis viscosus
(Conn).—Cocci in pairs and chains.
Colonies circular and granular.
Inoculated in the depth of gela-
tine liquefaction begins at the upper
part of the needle track, and extends
until the gelatine is completely
liquefied. The liquefied gelatine is
viscous, and may. be drawn out in
long threads.
On the surface of agar they form
a white, shining layer.
In broth there is an abundant
growth, and a film on the surface.
They coagulate milk, producing
butyric acid, and giving it a bitter
taste. They were obtained from
bitter cream.
Micrococcus luteus (Cohn).—
Oval cocci 1 to 1:2 » in diam.
Colonies yellow, with irregular
contours, and granular.
Inoculated in the depth of gela-
tine a granular filament develops in
the track of the needle, and on the
free surface a yellow patch.
On agar the growth is slimy and
yellow.
On potato the growth is yellow,
and after a time wrinkled.
The pigment is insoluble in water,
ether and alcohol, and not destroyed
by acids or alkalies.
They occur in water.
Micrococcus luteus (Schriter).
—Cocci similar in size to the above,
- gam).—Cocci
557
elliptical, with highly refractive cell
contents.
They form yellow drops of 1 to
3mm. diam. on boiled potato ; and
a thick, wrinkled, yellow skin on
nutrient liquids. :
The colouring-matter is insoluble
in water, and unchanged by sul-
phuric acid or alkalies.
Micrococcus ochroleucus
(Prove).—Cocci ‘5 to ‘8 y in diam.,
singly, in pairs, and short chains.
Colonies minute and colourless,
with crenated margin, from which,
later, processes extend into the gela-
tine, while the centre of the colony
becomes yellow.
On the surface of gelatine a film
develops, which in afew days turns
yellow. Old cultures have a pecu-
liar smell.
The yellow pigment can be ex-
tracted with alcohol. It is insoluble
in water, and decolorised by acids.
They were obtained from human
urine.
Micrococcus
472).
Micrococcus plumosus (Brauti-
‘8 » in diam., in
of Forbes (p.
masses.
Colonies yellowish-white.
Inoculated in the depth of gela-
tine long delicate acicular processes
‘shoot out from the needle track
and on the free surface.
On potato the growth is similar.
They were isolated from water.
Micrococcus pneumoniz crou-
pose (p. 236).
Micrococcus pyogenes tenuis
(Rosenbach).—Cocci irregular,
somewhat larger than staphylococci,
and with much less tendency to
form masses. The ends colour
deeply, leaving a clear space in the
middle.
Inoculated in the depth of gela-
tine a slightly opaque growth is
formed.
On agar a thin deposit appears
along the needle track, which is
almost as clear as glass.
They occur in the pus of un-
opened abscesses, but not often, as
they were found by Rosenbach only
in three out of thirty-nine cases.
558
Micrococcus rosettaceus (Zim-
mermann).—Cocci ‘7 to 1 win diam.,
singly, and in masses.
Colonies circular,
greyish-yellow.
Inoculated in the depth of gela-
tine the growth is very scanty in
the track of the needle, but
spreads over the surface as a grey
rosette.
On agar a smooth layer with den-
tated margin is formed.
On potato the growth is faintly
whitish, or
yellowish.
They occur in water. ;
Micrococcus roseus (Hisen-
berg).-—Cocci forming pink colonies,
and a rose-coloured growth on the
surface of nutrient agar-agar.
They were found in sputum ina
case of influenza.
Micrococcus salivarius septi-
cus (Biondi).—Oval cocci, diplo-
cocci, in chains and small masses ;
capsulated.
They grow best on acid gelatine,
or in an atmosphere of carbonic
acid.
Colonies are small and circular,
with an opalescent centre and a
transparent margin, with sharply
defined outline. In the interior of
the colonies there is an appearance
of a network.
Inoculated in the depth of gela-
tine the cocci form a delicate fila-
ment and white dots on the free
surface.
On agar the cultures should be
made direct from the blood. The
growth appears on the surface and
resembles dewdrops.
Broth cultures remain clear.
They are fatal to mice in twenty-
four to seventy-two hours, and to
rabbits in fifteen to thirty days,
producing septicemia. Attenuated
cultures are said to give immunity.
They were found in the saliva of
healthy and diseased persons.
Micrococcus stellatus (Mas-
chek).—Cocci singly.
Colonies stellate.
Inoculated in the depth of gela-
tine a branching growth appears in
the track of the needle. The jelly
becomes brownish.
DESCRIPTION OF SPECIES.
On potato the growthis brownish-
yellow and shining.
They occur in water.
Micrococcus tetragenus
(Gafikey).—Cocci about 1 p» in
diam., in tetrads, and surrounded
by a hyaline capsule.
Colonies form in twenty-four to
forty-eight hours as small white
dots, which are finely granular, and
have a vitreous lustre ; when they
reach the surface they form thick
raised masses.
Inoculated in the depth of gela-
tine the cocci form an irregular
white growth, especially in the
upper part of the track of the
needle.
On agar the colonies occur along
the needle track, and are white,
round and circumscribed.
On potato they form a thick,
slimy, viscous layer.
White mice inoculated with a
minute quantity of a pure-cultiva-
tion die in from two to ten days,
and the groups of the characteristic
tetrads may be found in the capil-
laries throughout the body, especi-
ally in the spleen, lung and kidney.
Double infection can be produced
by inoculating a mouse with a
pure cultivation of Bacillus an-
thracis two or three days after
inoculation with Micrococcus tetra-
genus. On examination after
death, the capillaries of the lungs,
liver and kidney are filled with
both anthrax bacilli and masses of
tetrads (Plate V., Fig. 3).
Micrococcus tetragenus
mobilis ventriculi (Mendoza).—
Cocci in tetrads; capsulated ;
motile. :
Colonies circular, whitish and
granular.
Inoculated in the depth of gela-
tine they grow on the free surface
only, and give off a peculiar odour.
They were isolated from the
stomach.
Micrococcus tetragenus sub-
flavus (Von Besser).—Cocci singly,
and in tetrads.
They do not grow on gelatine.
Colonies on agar are brown and
irregular in contour.
DESCRIPTION OF SPECIES.
_ On the surface of agar the cocci
form a greyish-white band, which
turns brown at the periphery, and
later is all dark or orange-yellow.
On potato the growth is brown.
They occur in nasal mucus.
Micrococcus tetragenus ver-
satilis (Sternberg and Finlay).—
Cocci varying in size from ‘5 to 1-5
#, in tetrads and irregular groups.
Colonies are circular and lemon-
yellow in colour.
Inoculated in the depth of gela-
tine there is very scanty develop-
ment along the line of puncture,
and the gelatine is liquefied in the
form of a cup near the surface.
At the bottom of the liquefied
gelatine a viscid, pale-yellow mass
accumulates.
On the surface of agar a thick,
viscid, yellow layer is formed along
the line of inoculation, which gradu-
ally extends over the entire sur-
face. The colour varies from cream-
yellow to lemon-yellow, and the
surface is moist and shining.
On potato there is a similar
growth. ,
They were isolated from the
skin of patients suffering from
yellow fever, from mosquitoes after
attacking these patients, and from
the air.
Micrococcus ure liquefaciens
(Fliigge).—Cocci spherical, 1:25 to
2 w» in diam., singly, or in chains
of three to ten elements, or in
irregular groups.
Colonies appear in two days
as small white points. They have
sharply defined edges anda granular
surface. The gelatine gradually
liquefies, and the edges of the colo-
nies become irregular.
The cocci inoculated in the depth
of gelatine produce a continuous
white line along the track of
the needle. Finally, the whole of
the gelatine liquefies, and appears
as a whitish-turbid fluid with a
thick whitish-yellow deposit at the
bottom.
They were obtained from urine.
Micrococcus versicolor
(Fliigge).—Small cocci, in pairs
and in masses.
» 559
White colonies develop in twenty-
four hours ; after two days they are
yellowish, with sharp contours of
yellowish-green colour, and finely
granular. The superficial colonies
form flat deposits, 2-6 mm. in size,
increasing to 10 mm. after four or
five days.
On the surface of gelatine the
cocci form a shining layer with a
greenish or bluish shimmer like
mother-of-pearl.
Inoculated in the depth of gela-
tine the growth is composed of
spherical yellowish colonies, and
on the free surface they form an
iridescent film.
They occur in the air.
Micrococcus violaceus (Schré-
ter).—Cocci or elliptical cells, de-
scribed as uniting into violet-blue
gelatinous spots, which again unite
to form larger patches.
The colonies on gelatine are violet
in colour.
Inoculated in the depth of gela-
tine the growth is scanty in the
track of the needle.
On the surface of gelatine they
form a bluish-yiolet layer, and the
same on agar and potato.
They were observed on boiled
potatoes exposed to the air, and are
also found in water.
Micrococcus viticulosus (Katz).
—Oval cocci 1 » in width, and 1:2 p
in length, in masses, but without
formation of much gelatinous ma-
terial.
The superficial colonies are quite
different in appearance from the
deep colonies. From the deep
colonies fine hairlike tendrils are
thrown off from a centre, forming
a very delicate and extensive net-
work. The threads are found to
consist of zooglcea masses, irregular
in size, arranged like strings of
beads. The colonies which are ex-
posed to the air form a thin layer.
of muddy-white gelatinous sub-
stance, which rapidly spreads, some-
times sending out hairlike processes
into the depth of the gelatine.
Inoculated in the depth of gela-
tine a delicate feather-like growth
occurs along the track of the needle,
560 DESCRIPTION
and on the free surface they pro-
duce the appearance which has been
described in colonies. This micro-
organism is exceedingly rare. It
was obtained from a contaminated
culture.
Monas Okenii,—Shortcylindrical
cells, 5 » wide, 8 to 15 » long, with
rounded ends. They exhibit lively
movements, each end being provided
with a flagellum twice as long as
the cell itself. They have pale-red
cell-substance, with dark grains.
They occur in stagnant water.
Monas vinosa.—-Round or oval
cells of about 2°5 » in diam., often
united in pairs. Their motion is
slow and tremulous, and the cell-
substance is pale-red with dark
grains interspersed. Flagella have
not been observed.
They were found in water with
decaying vegetable matter.
Monas Warmingii.—Cylindrical
cells, rounded at the ends, 15 p long,
5 to 8 wu broad. They are possessed .
of a flagellum at each end, and
exhibit rapid, irregular movements.
The cell-substance is pale-red, inter-
spersed at the ends with dark-red
grains.
Myconostoc gregarium (Cohn).
—The threads are very thin, colour-
less, unarticulated, but fall apart
into short cylindrical links when
dried.
They form gelatinous masses,
10 to 17 » in diam., singly or
heaped into slimy drops on water
in which alge are decomposing.
Nitromonas of Winogradsky.—
Very short rods, ‘9 to 1 » in width,
1:1 to 18 » in length. Singly, in
masses, and in very short chains.
They can be cultivated in silica-
jelly.
They are active agents of nitrifi-
cation.
They were obtained from the soil.
Pediococcus acidi lactici
(Lindner).—Cocci ‘6 to 1 in diam.,
singly, in pairs, and tetrads.
Colonies colourless.
On the surface of agar the cocci
form a colourless layer.
On potato the growth is almost
invisible.
OF SPECIES.
The cocci produce lactic acid in
solutions containing sugar.
They occur in hay infusion and
malt.
Pediococcus cerevisiz (Balcke).
—Cocci singly, in pairs, and tetrads.
Colonies at first colourless, later
yellowish-brown.
Inoculated in the depth of gela-
tine a greyish-white filament
occurs in the track of the needle,
and a white layer on the free
surface.
On agar the growth is transparent
and iridescent, and on potato almost ;
invisible.
They were isolated from the air
of a brewery.
Pneumobacillus
bovis (p. 242).
Proteus capsulatus septicus
(Banti).—Rods isolated from a case
of septicaemia, and identical with
Proteus hominis capsulatus.
Proteus hominis capsulatus
(p. 224).
Proteus in gangrene of the
lung (Babés).—Rods ‘8 to 15 p
thick, irregular in form, and fila-
ments with irregular enlargements.
Colonies whitish and transparent,
with ramifications extending over
the gelatine.
In the depth of gelatine a growth
occurs along the track of the
needle, and a ramifying growth on
the free surface.
On agar the growth is slightly
yellowish.
On potato the growth is brownish.
They are extremely pathogenic
in mice and guinea-pigs.
They were isolated from a case
of gangrene of the lung.
Proteus microsepticus (Kar-
linski).—Cocci, rods and filaments
in morphology, and cultures re-
sembling Proteus vulgaris.
Inoculated in the depth of gela-
tine liquefaction occurs in the
track of the needle, forming a
funnel with cloudy contents, and
in a few days the whole of the
gelatine is liquid.
They produce a general infection
in mice, and death in twenty-four
hours, and occasionally death in
liquefaciens
DESCRIPTION OF SPECIES. 561
rabbits, and local suppuration in | concentric circles, which in time
guinea-pigs and white rats. liquefies the medium. Similar
They were isolated from pus in | movements are observed in capsule-
a fatal case of puerperal pyeemia. cultivations as in Proteus vulgaris.
They were isolated from
putrid meat infusion.
Proteus septicus
(Babés).—Rods “4 yw in
width, and filamentous
forms.
Colonies rapidly lique
the gelatine. wg peas
Inoculated in the depth
of gelatine the bacilli form
a turbid funnel, or com-
pletely liquefy the gelatine
in twenty-four hours.
On agar the growth is
reticulated.
On potato brownish-
white.
Cultures have an un-
pleasant odour.
They are pathogenic in
mice.
They were isolated from
the organs in a case of
human septiczemia.
Proteus sulfureus (Lin-
denborn).—Rods -8 » in
Fig. 221.—Prorrus Murasiis: SwarMine width, varying in length,
ISLANDS ON THE SURFACE OF GELATINE, X 285 and long filaments. —
(Hauser). They correspend in mor-
phology and cultures with
Proteus mirabilis.—Cocci -4 » | Proteus vulgaris.
to’9 uw. They occur singly and in | They produce sulphuretted hy-
zoogloea, and sometimes in tetrads, | drogen in cultures.
pairs, chains, or as short. rods in They were isolated from water.
twos resembling Bacterium termo— Proteus vulgaris (Hauser).—
Fic. 222.—Prorevs Mrrasitis: InvorvTion Forms, x 524 (HAUSER).
in fact, in all conceivable transition Rods varying in size; some mea-
forms. sure 4 » in length, and are almost
Cultivated on nutrient gelatine | as broad as long, and others vary
they form a thick whitish layer in | from ‘94 to 1:25 » long and ‘42 to
36
562
63 p
motile.
Cultivated on nutrient gelatine
they convert it into a turbid, grey-
ish-white liquid. If cultivated in
a capsule containing 5 per cent. of
nutrient gelatine, a few hours after
inoculation the most characteristic
movements of the individual bacilli
wide. They are actively
fs oA
BN G s)
he : aie
oe is ann
@ oy Airy Wy — ie i
ay” ses a, u
y Ee Bi ANG
as ad wy il ) Wwe
Ma
Fic. 223.—Proreus VuLcanris,
Surrace oF NUTRIENT GELATINE, x
(Hauser).
FROM
are observed on the surface of the
nutrient gelatine, although at this
early stage no superficial liquefac-
tion can be detected. Probably the
movements depend upon the exist-
ence of a thin liquid layer, as they
are not observed if the nutrient
medium contains 10 per cent. of
gelatine.
They were isolated from putrid
meat infusion.
Proteus Zenkeri.—Cocci -4 », in
twos like Bacterium termo, and
short rods 1°65 p long.
Cultivated on nutrient gelatine
no liquefaction results, but a thick
whitish-grey layer is formed. The
THE
.DESCRIPTION OF SPECIES.
bacilli are motile, and the same
phenomena are observed on the
solid medium as in Proteus vulgaris.
In cover-glass impressions most
varied groupings of the bacilli are
seen, and also developmental and
involution-forms.
They were isolated from putrid
meat infusion.
Pseudo-diphtheritic bacil-
lus (p. 335).
Pseudo-diplococcus pneu-
moniz (Bonome).—Oval cocci
in pairs and short chains ; cap-
sulated.
Inoculated in the depth of
gelatine small colonies develop
in the track of the needle in
twenty-four hours.
On agar there is a scanty,
moist growth.
On potato an almost in-
visible film.
In broth the cocci grow
rapidly, and the cultures give
off a peculiar odour.
They produce septicemia in
mice, guinea-pigs and rabbits.
This micro-organism is
probably a variety of the
pneumococcus.
They were isolated from a.
fatal case of cerebro-spinal
meningitis.
Rhabdomonas _ rosea. —
Spindle-form rods, 3°8 to 5 p»
broad, 20 to 30 w long. They
exhibit slow, trembling move-
ments, having at each end of
the cell a flagellum. The
cell-substance is very pale, with
dark grains interspersed.
They occur in brackish water.
Sarcina alba—Small cocci.
They form small white colonies on
nutrient gelatine.
Inoculated in the depth of gela-
tine they grow slightly along the
needle track, but are heaped up on
the surface without liquefying the
gelatine.
They are present in the air.
Sarcina aurantiaca,—Cocci
singly, in pairs, in tetrads, and in
packets.
Colonies orange-yellow.
Inoculated in the depth of
285
‘DESCRIPTION
gelatine they slowly liquefy it along
- the whole needle track, and form
on the surface an orange-yellow
growth. On potatoes they slowly
develop the same pigment.
Sarcina candida (Reinke).—
Cocci 1:5 to 1°7 » in diam., singly,
in pairs, and in tetrads.
Colonies are circular and shining,
white, and later yellowish.
Inoculated in the depth of gela-
tine liquefaction quickly takes place
along the track of the needle.
On the surface of agar a white,
moist layer develops.
They were found in the air of
breweries.
Sarcina flava (De Bary).—Small
cocci in packets.
Inoculated in the depth of gela-
tine they produce liquefaction.
On agar they form a yellow
layer.
On potato the growth is limited
and yellow.
They were isolated from beer.
Sarcina hyalina (Kiitzing).—
Cocci round, 2°5 » in diam., almost
‘colourless. United in families of
4 to 24 cells, reaching 15 p in diam.
They occur in marshes.
Sarcina intestinalis (Zopf).—
Cocci in groups of four or eight.
Very regular in form; never in
the large packets which occur in
-Sarcina ventriculi.
They are found in the intestinal
canal, especially the cecum, of
poultry, particularly fowls and
turkeys.
_ Sarcina litoralis (Oersted).—
Cocci 1-2 to 2 w in diam., bound
together in 4 to 8 families, which,
in their turn, may unite and in-
clude as many as 64. tetrads.
-Plasma colourless; in each cell
1 to 4 sulphur granules.
They were found in sea water
containing putrefying matter.
Sarcina lutea (Schréter).—
Cocci singly, in pairs, tetrads and
packets. <A single individual in a
‘tetrad may be divided into two, or
into four, so that a tetrad within a
tetrad results.
Colonies are round, slightly.
granular in appearance, and yellow.
OF SPECIES, 563.
Inoculated in the depth of
gelatine they grow rapidly; the
gelatine becomes liquefied, and the
yellow growth sinks to the bottom
of the tube.
Greys
Saeer 2
dings, int
RZ O Sree
enh EP ihe, ‘a SB
SEC! aay,
Gis re CkOe C hel
Fic. 224,—Sarcina x 600 (Fiiiccr.
Cultivated in agar they form a.
colourless growth along the track
of the needle, and a bright canary-
yellow layer upon the surface.
On potato they form a yellow
layer.
They are present in air.
Sarcina mobilis (Maurea),—
Cocci 1°5 » in diam., in pairs, and
in tetrads. They are motile.
Colonies, at first white, become:
brick-red. ;
Inoculated in the depth of gela-
tine, there is, after several days, a.
slight growth along the track of
the needle, and a patch of growth
on the free surface which gradually
turns-red. In about two weeks.
liquefaction produces a_ funnel-
shaped appearance ; later the lique--
faction extends to the sides of the
test-tube.
In broth turbidity is produced,.
and a yellowish-red deposit.
On agar the growth, at first white,
‘changes to a brick-red colour.
There is no growth on potato,.
and milk is not coagulated.
They were isolated: from ascitic.
fluid. ;
Sarcina pulmonum (Hauser).—
Cocci from 1 to 1°5 » in diam., in
tetrads and packets.
Colonies white and small. They
are coarsely granular.
Inoculated in the depth of gela-
tine the growth is scanty in the
track of the needle, but on the free
surface there is a circular, well-
‘defined, translucent patch, which
564 DESCRIPTION OF SPECIES.
later becomes greyish-brown, shin- -
ing, wrinkled and irregular.
_On potato the growth is very
slight and limited.
They cause ammoniacal decom-
position of urine.
They were isolated from phthisi-
cal sputum.
Sarcina Reitenbachii (Caspary).
—Cocci about 1:5 to 2°5 » in diam.,
at the time of division lengthened
to 4 pp. Mostly united together
from 4 to 8 in number ; occasion-
ally 16 or more. Colourless cell-
wall, lined with rose-red layer of
plasma.
‘They were found on rotting
water-plants.
Sarcina rosea (Schroter).—
Large cocci, in packets.
Inoculated in the depth of gela-
tine, liquefaction quickly takes
place, and cultures after a time
have a reddish colour.
On agar the growth is slow and
limited.
On potato the growth is abundant
and of a bright-red colour.
In broth they produce turbidity,
and a red deposit. :
They occur in the air.
Sarcina urine (Welcker).—Very
small cocci, 1:2 » in diam., united
in families of 8 to 64. They were
observed in urine.
Sarcina ventriculi (Goodsir).—
‘Cocci reaching 4 » in diam., united
in groups of four, or multiples of
four, producing cubes or packets
with rounded-off corners. Contents
of the cells are greenish or yellow-
ish-red.
Colonies are round and yellowish.
A yellowish growth forms on the
surface of oblique gelatine without
liquefaction.
On potato they form a yellow
growth, and on serum also.
They grow well in hay-infusion,
forming brownish scales and a simi-
larly-coloured deposit.
They occur in the stomach of man
and animals in health and disease,
and were first detected in vomit.
Spherotilus natans.—Cells 4 to
9 p» long, and 3 p thick, united in
a gelatinous sheath to form threads.
The cells comprise rods and cocci-
forms ; the cocci are set free, and .
develop into rods, which again form
threads. In the last a false branch-
ing has been observed. The plasma
of the cells break up into minute,
strongly-refractive portions, which
develop into round spores, at first
of a red and afterwards a brown
colour.
They occur in stagnant and flow-
ing water contaminated with or-
ganic matter, and form floating
flakes of a white, yellow, rust-red,
or yellow-brown colour.
Spirillum amyliferum (Van
Tieghem).—Filaments 6 » in length
and 1:4 to 1°5 w in width; with
from 2 to 4 screw curves.
They act as a strong ferment in
the absence of air.
They occur in water.
Spirillum anserum (Sakharoff),
—Spirilla resembling the Spiro-
cheta Obermeieri Extremely
motile.
They have not been cultivated
artificially.
Blood from diseased geese, con-
taining the spirilla, produces the
disease when inoculated in healthy
birds. The geese suffer from
diarrhoea, and die in about a week.
They were found in the blood of
geese suffering from an epidemic
form of septicemia prevailing at
some of the stations on the Trans-
caucasian Railway.
Spirillum attenuatum (Warm-
ing).—Threads much attenuated at
the ends, which consist usually of
three spirals. The middle spiral is
about 11 » high and 6 » in diam.,
and the end ones 10 » high and 2 »
in diam.
They are found in brackish water.
Spirillum aureum (Weibel).—
Curved rods with blunt ends, spi-
rilla and spirilliform filaments, and
involution forms.
Colonies are circular and golden-
yellow.
Inoculated in the depth of gela-
tine, a finely granular growth forms
in the track of the needle, and a
yellow-ochre prominent mass on
the free surface.
-
DESCRIPTION
On agar a greyish growth ex-
tends over the surface, and later
prominent yellowish heaps make
their appearance.
On potato there is an abundant
growth of a golden-yellow colour.
They were found in sewage
mud.
sr ee cholere Asiatice
Spirillum choleroides (Buj wid).
—Curved rods very similar mor-
phologically and in cultures to
Koch’s comma-bacilli.
They were isolated from river
water.
Spirillum choleroides (Orlow-
ski)._Curved rods very similar to
Koch’s comma-bacilli.
They were found in well water.
Spirillum concentricum
(Kitasato)—Short spirilla, and
spirilliform filaments.
Colonies are circular, and com-
posed of concentric rings alternately
opaque and transparent.
Inoculated in the depth of gela-
tine there is only a little growth
in the track of the needle, and a
cloudy growth on the surface
extending into the jelly.
On agar the’ growth is extremely
adherent.
In broth they produce turbidity,
which disappears after a time ; and
there is a slimy deposit at the
bottom of the tube.
They were found in putrefying
blood. f
Spirillum dentium (Miller).—
Spirals 10 to 20 » in length, pointed
at the ends.
They have not been cultivated.
They occur in the deposit on the
teeth, and in company with Lepto-
thrix buccalis in carious teeth.
Spirillum flavescens (Weibel).
—Commas thicker than those found
in Asiatic cholera, spirilla, and
spirilliform filaments.
Colonies yellowish.
Inoculated in the depth of gela-
tine a finely granular filament
develops in the track of the needle,
and on the free surface a pale-
yellow patch.
On agar the growth, at first
OF SPECIES. 565
greyish-white, becomes yellow, and
forms a thick layer.
On potato the growth is abun-
dant, and similar in colour.
They were found in
mud.
Spirillum flavum (Weibel).—
Spirilla morphologically identical
with spirillum aureum.
Colonies on gelatine are pale-
yellow, and later the colour is more
intense.
On agar and potato they forma
layer the colour of yellow-ochre.
They were isolated from sewage
mud.
Spirillum leucomelaneum
(Koch).—Dark and glass-like spaces
alternate in the spirillum, resulting
from a regular arrangement of the
dark granular contents. A rare
form observed in water covering
rotting algze.
Spirillum linguze (Weibel).— -
Curved rods, spirilla, and spirilli-
form filaments, and involution
forms.
Colonies are composed of inter-
lacing filaments, and offshoots
extend into the surrounding gela-
tine.
Inoculated in the depth of gela-
tine a delicate growth occurs in the
needle track.
On agar the growth is whitish
and granular.
In broth a cloudiness is produced,
‘as well as a flocculent deposit.
They are especially distinguished
from other spirilla described by
Weibel by their staining by Gram’s
method.
They were isolated from the
tongue. :
Spirillum marinum (Russell).
—Curved rods, and spiral filaments.
Colonies circular, granular and
striated ; later, flocculent masses
float in liquefied areas.
Tnoculated in the depth of gela-
tine liquefaction occurs in the
track of the needle, and a mem-
brane forms on the surface of the
cloudy liquid. ;
On agar the growth is yellowish
and abundant.
On potato a thick, waxy mass
sewage
566
develops, and extends over the
surface.
Broth made with sea-water be-
comes rapidly turbid.
They were obtained from sea-
water.
Spirillum Metchnikovi (p. 373).
Spirillum nasale (Weibel).—
Large curved rods and spirilliform
filaments. Non-motile.
Colonies circular, finely granular,
and brownish-yellow.
Inoculated in the depth of gela-
tine a delicate growth develops in
the track of the needle.
On the surface of agar they form
a whitish slimy film.
They occur in nasal mucus.
Spirillum Obermeieri (p. 258).
Spirillum of Finkler and
Prior (p. 258).
Spirillum of Giinther (Vibrio
aquatilis)—Curved rods, very simi-
lar to Koch’s comma-bacilli, but
there is no growth on potato.
They occur in water.
Spirillum of Miller—Curved
rods, singly, in pairs, and spiral
filaments.
They liquefy gelatine.
They were isolated from carious
teeth.
Spirillum of Neisser (Vibrio
berolinensis)—Similar to Koch’s
comma-bacillus, but smaller.
Colonies colourless, granular and
transparent, liquefying the gelatine
much more slowly than Koch’s
comma-bacillus.
Inoculated in the depth of gela-
tine they produce a growth similar
to that of Koch’s comma-bacilli,
but much more slowly in milk.
In broth they grow abundantly.
They were found in water.
Spirillum of Rénon.—Curved
rods longer and broader than the
comma-bacilli of Koch.
Colonies yellowish, with dark
nucleus.
In gelatine the cultures resemble
those of Koch’s comma-bacillus.
On agar the growth is white and
abundant.
They cause turbidity in broth.
They were isolated from impure
water from a well.
DESCRIPTION OF SPECIES,
Spirillum of Smith (vide
Spirillum suis).
Spirillum of Weibel —Curved
rods resembling Koch’s comma-
bacilli, morphologically, and in
cultures on jelly.’ The gelatine is
more quickly liquefied.
There is no growth on potato.
They occur in well water.
Spirillum plicatile (Ehrenberg).
—Thin threads 2°25 » in breadth,
110 to 125 p» long, occurring also in
spirulinar forms. The threads have
primary and secondary windings ;
the former are in each example of
equal size, but the latter are often
Fic 225.—Sprocua#ra PLicaTite.
irregular. Their ends are cut off
bluntly, and they exhibit rapid
movement.
They occur abundantly in marsh-
water in summer, and can be ob-
tained by allowing alge to decom-
pose in water. On cultivation the
threads break up into long rods,
short rods, and, finally, cocci. This
change is rendered visible by making
cover-glass preparations, and stain-
ing with aniline dyes.
Spirillum rosaceum (Klein).—
Resembles Spirillum undula, but is
reddish in colour ; the colouring-
matter is insoluble in water, alcohol
or chloroform.
Spirillum Rosenbergii—
Threads with 1 to 14 windings, 4
DESCRIPTION OF SPECIES.
to 12 p» long, 1:5 to 2°6 p» thick. |
They are colourless, but the con-
tents include strongly refractive
sulphur granules. Also spirals 6 to
75 » in height, which are actively
motile.
They were found in brackish
water.
Spirillum rubrum (Esmarck).
—Curved rods, spirilla and spiro-
chetz. They are actively motile.
The growth on artificial media is
extremely slow.
Inoculated in the depth of gela-
tine they grow along the track of
the needle, forming a filament of a
wine-red colour, without causing
liquefaction ; and on the free sur-
face the growth is colourless.
In broth long spirillar threads
are formed.
They were isolated from the
putrid tissues of a mouse.
Spirillum rufum (Perty).—
Filaments from 8 to 16 » in length,
with 13 to 4 screw curves; non-
segmented ; chiefly motile ; tinged
with red.
They form rose or dark red spots
on the sides of wells.
Spirillum sanguineum (Cohn).
—Threads 3 » and more in thick-
ness, with 2 to 24 spirals, each 9 to
12 phigh. The ends are provided
with flagella. Their colour is due
to the presence of reddish granules
contained in the cells.
They were observed in brackish
water containing putrefying sub-
stances. (Vide Beggiatoa roseo-
persicina. )
Spirillum _ saprophiles.—(..)
Curved rods with pointed ends, ‘6 p
in width, 3 p» in length; spirilla,
spirilliform filaments, and involu-
tion forms.
Colonies yellowish or greenish-
yellow.
Inoculated in the depth of gela-
tine a white growth forms in the
track of the needle, later becoming
yellowish ; and on the free surface
there is a white growth, and
beyond this a transparent film
spreads over the jelly.
On agar the growthis creamy,
and the jelly clouded beneath it.
567
‘On potato the growth is slimy
and yellowish or dark-brown in
colour, :
They were obtainéd from sewage
Fig. 226.—ComMa-BAcILLI IN WATER
CONTAMINATED WITH SEWAGE.
mud and decomposing hay infu-
sion.
CII.) Curved rods about 2 » in
length, with blunt ends and in pairs.
Extremely motile.
Colonies circular and yellowish-
brown.
Inoculated in the depth of gela-
tine a white growth develops in
the track of the needle, and later
becomes yellowish-red ; on the free
surface a white patch forms, sur-
rounded by a transparent film.
In the depth of agar there is no
growth in the track of the needle,
but a yellowish-white patch on
the free surface adherent to the
jelly.
On potato the growth is also
adherent, and in appearance shining
and brownish-green.
They were isolated from decom-
posing hay infusion. —
‘(II.) Curved rods, spiriila, and
spirilliform filaments, and involu-
tion forms.
Colonies are circular, granular,
and with irregular margin ; yellow
in the centre, and white at the
| periphery.
Inoculated in the depth of gela-
tine a white growth develops in
‘the track of the needle, and on the
_ surface, without producing lique-
faction.
On the surface of agar the growth
is white.
. On potato the growths distinctly
brown.
568
They were isolated from sewage
mud.
Spirillum serpens (Miiller).—
Long spirilliform filaments ; often
collected in masses.
They were observed in vegetable
infusions and stagnant water.
Spirillum sputigenum (Lewis).
—Curved rods, very similar to the
comma-bacilli of Koch ; but many
observers having failed in repeated
Wes.
sy
Bas Ss
BENGE oO
EM
AM ae Ie
ON. 2 wt i
eA y
Kg FEF
Si
Fie. 227.—Comma-BacILLI OF THE
Mourn, x 700 (Van ERMENGEM).
attempts to cultivate them, main-
tain that they are biologically dis-
tinct from those associated with
Asiatic cholera. Klein asserts that
they can be cultivated in an acid
gelatine, and that they are iden-
tical with Koch’s comma-bacilli in
their mode of growth. They
occur with other bacteria in saliva
and in scrapings from carious
teeth.
Spirillum suis (Smith).—Com-
mas and spirilla.
Colonies in gelatine are circular,
granular and brownish, and later
appear to be composed of concen-
tric rings. The gelatine is not
liquefied.
Broth with 1 per cent. of peptone
becomes in a few days clouded.
On potato they develop a thin
yellowish layer. :
larger than those obtained from
Asiatic cholera, and are
pathogenic.
They were obtained from the
large intestine in swine.
Spirillum tenue.—Very thin
threads, with at least 13, usually
2 to 5 spirals. Height of a single
screw is 2 to 3 yw, and the length of
spiral therefore 4 to 15 p. They
are very swiftly motile.
not |.
DESCRIPTION OF SPECIES.
They often occur in dense felted
swarms in vegetable infusions.
Spirillum tyrogenum (Deneke).
—Curved rods, slightly smaller
than Koch’s comma-bacilli, with
a great tendency to form long
spirillar threads (Fig. 228).
il k
s SNe = at
o ~ =~
~ ay |
~%) an as =
ee =
se is” (s/7 a?
Shee a
. aE eR
~—
~
Fic. 228.—Denexe’s ComMma-BaciLitr
FROM CHEESE, x 700 (FitiecE).
Colonies on _plate-cultivations
are sharply defined and of a
greenish-brown colour. After a
time they liquefy the gelatine, but
the liquefaction is much more
marked than in colonies of Koch's
commas of the same age, though
not so rapid as in the case of the
commas of cholera nostras.
Inoculated in the depth of nu-
trient gelatine a turbid liquefaction
occurs along the needle track, and
on the surface of nutrient agar-agar
a yellowish-white layer develops.
Inoculation of potatoes gives no
result.
Administration of the bacilli by
the mouth, in the manner employed
for testing the pathogenic effect of
Koch’s bacilli, produced a fatal
result in a few cases ; on the other
| hand, injection into the duodenum
| failed entirely.
. properties may be therefore con-
‘| sidered as not yet established.
The commas are said to be slightly |
The pathogenic
| They were isolated from old
cheese.
Spirillum undula.—Threads
11 to 1:4» thick, 9 to 12 » long;
: spirals 4°5 » high ; each thread has
' 1$ to 3 spirals.
‘motile, and possess a flagellum at
, each end.
They are actively
They occur in various infusions.
Spirillum volutans (Ehrenberg).
_ Threads 1:5 to 2 » thick, 25 to
30 » long, tapering towards their
DESCRIPTION
extremities, which are rounded off.
They possess dark granular con-
tents. Each thread has 23 to 34
windings or spirals, whose height is
9to13 ». They have a flagellum
at each end, and are sometimes
motile, sometimes not.
They are found in the water of
marshes and in various infusions.
Spiromonas Cohnii—Colourless
cells, consisting of 14 spirals, with
both ends acutely pointed and pro-
vided with a flagellum. Breadth
of the cells, 1:2 to 4 p.
They occur in water containing
decomposing matter.
Spiromonas volubilis (Perty).
—Colourless, transparent cells, 15
to 18 » long. Rapidly motile, and
revolving round a longitudinal
axis.
They occur in marsh-water and
putrefying infusions,
Staphylococcus pyogenes albus
(p. 178).
Staphylococcus pyogenes
aureus (p. 176).
Staphylococcus pyogenes cit-
reus (p. 178),
Staphylococcus _pyosepticus
(Heucourt and Richet).—Cocci
identical with Staphylococcus pyo-
genes aureus.
Subcutaneous injection causes in
rabbits intense cedema, and death
in twenty-four hours.
They were isolated from pus
from an abscess in a dog.
Staphylococcus salivarius pyo- |.
genes (Biondi).—Cocci 3 to ‘5 p
in diam., singly and in masses.
Colonies white and opalescent,
producing liquefaction.
Inoculated in the depth of gela-
tine the growth appears in the
track of the needle, and is followed
by liquefaction.
On agar the growth is orange-
yellow.
The cocci produce local suppura-
tion when inoculated in animals.
They were
subcutaneous injection of saliva.
This coccus is probably identical |
with Staphylococcus pyogenes |,
aureus.
isolated from an |
abscess in a guinea-pig following |
OF SPECIES. 569
Staphylococcus viridis flaves-
cens (Guttmann).—Cocci singly, in
pairs and masses ; morphologically
agreeing with Staphylococcus pyo-
genes aureus.
Colonies are greenish-yellow.
Inoculated in the depth of gela-
tine a filament forms composed of
greyish colonies.
On agar the growth is greenish-
yellow.
They grow well on potato.
They were isolated from the
vesicles of chicken-pox.
Streptococcus acidi lactici
(Grotenfeld).—Oval cocci ‘5 to 1 p
long, ‘3 to *6 » in width, and long
chains. They are partially anaerobic.
Colonies are circular and white.
Inoculated in the depth of gela-
tine a growth occurs only in the
track of the needle.
Milk is coagulated.
They were isolated from coagu-
‘lated milk.
Streptococcus albus (Tils).—
Cocci forming motile chains.
Colonies are flat and circular,
with white periphery and dark
nucleus, rapidly liquefying.
Inoculated in the depth of gela-
tine there is rapid liquefaction in
the track of the needle, and a
white deposit.
On potato they form a white
slimy layer.
They were found in water.
Streptococcus bombycis(p. 472).
Streptococcus brevis (Lingels-
heim).—Cocci_ singly, in pairs
and chains, of eight to ten
elements.
Colonies on gelatine are circular
| and very minute.
Inoculated in the depth of gela-
} tine there is a funnel-shaped cavity
| near the surface, and below this,
| in the track of the needle, small
| isolated colonies.
On agar a yellowish-grey film
| develops along the line of inocula-
tion. ; :
On potato there is a copious white
growth in forty-eight hours. -
Broth is made turbid.
They were isolated from healthy
| saliva.
570
Streptococcus cadaveris (Stern-
berg).—The description corresponds
with that of Streptococcus pyo-
genes. ;
¥{ Inoculated in the depth of gela-
tine the colonies are said to be
larger and more opaque.
On the surface of agar they form
a thin translucent layer.
In broth little flocculi develop,
composed of chains in which in
some cases the elements varied
considerably in size.
They were obtained from the
liver in a fatal case of yellow fever.
Streptococcus coli gracilis
(Escherich).—Cocci from ‘2 to “4 p
in diam., forming chains composed
of from six to twenty elements.
Some elements in a chain are irre-
gular in form, and show transverse
fission.
The colonies are spherical and
sink down in the liquefied gela-
tine.
Inoculated in the depth of gela-
tine liquefaction occurs in the track
of the needle on the second day,
and a white deposit forms at the
bottom of the liquid. In about a
week the gelatine is completely
liquefied.
On agar there is a very slight
growth.
On _blood-serum
develop.
On potato the growth is com-
posed of small white prominent
colonies.
Milk is coagulated.
They were found in the evacua-
tions of healthy infants.
Streptococcus conglomeratus
(ieurih)—Cood and chains, identi-
cal with Streptococcus pyogenes.
They form an adherent film at
the bottom of the tube, which is
not broken up by agitation. This
is observed in other varieties of
Streptococcus pyogenes, and is not
sufficient to distinguish it.
They are pathogenic in mice.
They were isolated from cases of
scarlet fever.
Streptococcus flavus desidens
(Fliigge).—Cocci, diplococci, and
short chains. They form yellowish-
small scales
DESCRIPTION OF SPECIES.
white colonies, which gradually
sink down in the gelatine.
Inoculated in the depth of gela-
tine the cocci form china-white,
confluent masses in the track of
the needle, and on the surface a
yellowish-brown slimy layer,
They occur in air and in water,
and were originally isolated from
contaminated cultures.
Streptococcus giganteus
urethre (Lustgarten).—Cocci
to 1 » in diam., forming chains
composed of several hundred
elements. In description they
correspond with Streptococcus
pyogenes.
They do not grow at the tem-
perature of the room.
Colonies on agar are transparent
and iridescent.
They were isolated from the
healthy urethra.
Streptococcus Havaniensis
(Sternberg).—Cocci -6 to "9 » in
diam., forming long chains, com-
posed of cocci, in pairs, and oval
elements showing transverse divi-
sion.
This streptococcus is probably
a variety of Streptococcus pyo-
genes.
They were found in the acid
vomit of a yellow-fever patient.
Streptococcus in contagious
mammitis in cows (Nocard and
Mollereau).—Cocci spherical or
oval, united in long chains.
Colonies are spherical, granular,
pale-yellow, or brownish by trans-
mitted light.
The cocci inoculated in the depth
of gelatine produce a granular
filament in the track of the
needle.
On the surface of nutrient gela-
tine minute spherical colonies are
formed, which are bluish by re-
flected light.
‘Injected into the mammary gland
of cows and goats they produce
mastitis.
They were isolated from the milk
of cows suffering from contagious
mammitis.
From the description this strepto-
coccus appears to be closely related
DESCRIPTION
to, if not identical with, Strepto-
coccus pyogenes bovis(Crookshank).
Streptococcus in progressive
tissue necrosis in mice—Koch
produced a disease in mice by sub-
cutaneous injection of putrid blood.
In tissue sections a chain coccus
was found which was similar to
Streptococcus pyogenes.
Fig. 229.—Srreptococcus Pro-
IN
GRESSIVE TissuE NEcROsIS IN MICcE.
a, Necrotic cartilage cells, and (b)
chains in masses; ¢, isolated chains.
(Koch.)
Streptococcus in Strangles
(Schutz).—Cocci forming long
chains, which, it is said, do not grow
on nutrient gelatine or agar, but
form a transparent iridescent cul-
ture on blood serum. Cultures in
broth produced the disease in horses
and mice. Rabbits, guinea-pigs,
and pigeons are not affected. ~
Strangles is a disease of the horse,
associated with suppuration of the
glands of the head and neck, prin-
cipally in the sub-maxillary, sub-
parotideal, and retro-pharyngeal
regions. Schutz found that the pus '
contains streptococci and produces
a fatal disease in mice.
Streptococcus liquefaciens
(Sternberg).—Spherical and oval
OF SPECIES. 671
cocci, ‘4 to 6 » in diam., singly, in
pairs and short chains.
Inoculated in the depth of gela-
tine liquefaction occurs rapidly in
the track of the needle, and in a
week the gelatine is completely
liquefied, slightly opalescent, and a
scanty deposit forms at the bottom
of the tube.
In the depth of agar a filament
is formed composed of closely-
crowded colonies,
On potato a thin and limited
dry white layer is formed along the
line of inoculation in four to five
days.
They are non-pathogenic.
They were isolated from the liver
and intestines of fatal cases of
yellow fever.
Streptococcus mirabilis (Ros-
coe and Lunt).—Cocci 4 yw in diam.,
singly, and in long chains.
The growth on nutrient media is
very scanty.
In broth the growth is composed
of a mass of delicate -filarnents
which collect at the bottom of the
liquid.
They were isolated from sewage.
Streptococcus of Bonome.—
Cocci forming chains. They corre-
spond in description with Strepto-
coccus pyogenes, but, it is said,
they do not grow in gelatine or
on blood-serum, and they are said
to be distinguished by the characters
of the colonies on agar plates.
They are pathogenic.
Inoculated in rabbits and white
mice they produce symptoms
similar to those produced by in-
oculations of the pneumococeus.
Sub-cultures rapidly lose their
virulence.
They were isolated from cases
of cerebro-spinal meningitis.
Streptococcus of Manneberg —
Cocci ‘9 » in diam., singly, in pairs,
and in chains.
Inoculated in the depth of gela-
tine a white filament forms along
the track of the needle composed
of minute colonies. In about a
month the filament is replaced by
a funnel of semi-liquefied gelatine.
On the surface of agar the
572
growth resembles Streptococcus
pyogenes.
On potato they form a slimy
layer.
Milk is rapidly coagulated.
They are pyogenic in dogs and
rabbits. Injected into the veins
they produce inflammation of the
kidneys.
They were isolated from the
urine in a case of Bright’s disease.
Streptococcus perniciosus
psittacorum (Parrot disease).—
Cocci, singly, in chains, and in
zoogloea, have been described in
connection with a disease of the
grey parrot (Psittacus erithacus).
This disease is fatal to about'80 per
cent. of these parrots imported to
Europe. They suffer from diarrhoea
and general weakness ; their feathers
are ruffled;
loosely, and their eyelids close ;
convulsions set in, and death fol-
lows.
nodules are found in the lungs, liver,
spleen and kidney. In and around |
the capillaries of these nodules, and
in the blood of the heart, the cocci |
are found in great numbers in |
zoogloea, and more rarely in chains.
Inflammatory change in the sur-
rounding tissue is absent.
Streptococcus pyogenes (p. 178). _
Streptococcus radiatus
(Fliigge).—Cocci less than 1 p in |
diam., singly, in small masses, and
sometimes in short chains.
Colonies appear in twenty-four .
hours. They are white, with a
yellowish-green sheen; later they
liquefy the gelatine and develop a
circlet of rays.
Inoculated in gelatine, isolated
centres form along the track of the |
needle which throw out horizontal |
rays. At the same time a funnel-
shaped area of liquefaction forms |
very slowly in the upper part.
On potato the growth is yel-
lowish-brown. :
They occur in air and in water.
Streptococcus septicus (Fliigge).
—Cocci in chains, indistinguishable
microscopically from Streptococcus
pyogenes.
Colonies on gelatine grow more
their wings hang |
At the autopsy greyish |
DESCRIPTION OF SPECIES.
slowly than those of most strepto-
cocci.
They are pathogenic, Mice die
in forty-eight to seventy-two hours
after subcutaneous inoculation of
a minute quantity of a cultivation.
During the last twenty-four hours
there is a distinct motor and sensory
paralysis of the hind legs. In
rabbits inoculation of the ear pro-
duces local redness, then a general
disease, and death in two or three
days.
They were found by Nicolaier,and
independently by Guarneri, in earth.
Streptococcus septicus lique-
faciens (Babés).—Cocci °3 to °4 p,
in pairs, in short chains.
Inoculated in the depth of gela-
tine a granular filament forms in
twenty-four hours along the track
of the needle, followed by lique-
faction of the gelatine forming a
funnel in which the gelatine is
clouded ;_ flat, whitish deposits
form on the side of the funnel.
On the surface of agar minute
shining, transparent colonies are
formed.
On blood-serum the growth is
almost invisible.
They are pathogenic. Subcu-
taneous injection in mice and
rabbits produces local inflammation
and cedema, followed by death in
about a week.
They were found in the blood
and organs of a child which had
died of septicemia complicating
scarlet fever.
Streptococcus | vermiformis
(Tils)—Cocci forming motile
chains. ;
Colonies are yellowish-white, the
central portion finely granular,
the periphery radiated.
Inoculated in the depth of gela-
tine there is rapid liquefaction,
and a yellowish deposit at the
bottom of the liquid.
On potato the culture forms a
dirty-yellow layer.
They were found in water.
Streptothrix actinomycotica
(p. 431).
Streptothrix alba (Gasparini).
—A variety of Actinomyces bovis.
DESCRIPTION
Streptothrix asteroides
(Oospora asteroides, Sauvageau
and Radais ; Cladothrix asteroides,
Eppinger). — Branching filaments
which form on the surface of
grape-sugar-agar a whitish growth,
which is later of a brownish-yellow
colour.
Broth remains clear, and small
pellicles float on the surface re-
sembling drops of stearin.
On potato they form snow-white
points, which turn brick-red in
colour, and are later covered with
a delicate white efflorescence.
The streptothrix is pathogenic
in rabbits and guinea-pigs.
It was isolated from pus.
Streptothrix aurantiaca
(Oospora aurantiaca, Sauvageau and
Radais, and Doria).—Similar to -
Streptothrix asteroides.
Streptothrix carnea (Doria ;.
Oospora carnea, Sauvageau and
Radais).—Similar to Streptothrix
asteroides, but the cultures on
gelatine are pink.
They are not pathogenic.
Streptothrix chromogenes
(Gasparini; Oospora chromogenes,
Lehmann and Neumann).—Culti-
vated on the surface of gelatine the
filaments produce a chalky growth,
and the jelly is coloured brown, and
is slowly liquefied.
\On potato the growth is yellowish
or brown, and the potato itself is
coloured dark brown or black.
The streptothrix has been isolated
from air and water and the con-
tents of the stomach.
Streptothrix farcinica (Bacille
du farein de beuf, Nocard ; Oospora
facinica, Sauvageau and Radais).
Inoculated on the surface of
gelatine there isin about two weeks
a very scanty granular growth.
In broth greyish pellicles develop
with a dusty surface. They are
pathogenic in cattle, guinea-pigs,
and sheep.
They were isolated from the
disease known as farcin de beuf.
Streptothrix Forsteri (Cohn).
—Cocei rods, and leptothrix threads.
The threads are twisted in irregular
spirals, and branch sparingly and
OF SPECIES. 573
irregularly. Screw-forms are pro-
duced by the threads breaking up
into small pieces.
Colonies slowly liquefy gelatine.
On agar they form a whitish
growth.
In broth they form shining
masses, floating in clear liquid.
They occur in the lachrymal
canals of the human eye, in the
“form of closely felted masses, and
in the air, and in fresh- and sea-
water.
Streptothrix Hofmanni (Jficro-
myces Hoffmanni, Gruber ; Oospora
Hoffmanni, Sauvageau and Radais).
—The filaments flourish in the ordin-
ary culture media with the addition
of sugar, but they do not grow on
potato.
They produce
rabbits. ~
They were isolated from the air.
Streptothrix liquefaciens
(Cladothrix liquefaciens, Garten).—
A variety of Actinomyces bovis.
Streptothrix madurae (p. 449).
Streptothrix musculorum
suis (Actinomyces suis, Dunker).
—A variety of actinomyces found
in the muscles of swine.
Streptothrix odorifera(Oospora
odorifera, Rullmann). Probably
identical with Oospora chromogenes.
Streptothrix violacea (Qospora
violacea, Sauvageau and Radais,
and Doria—This streptothrix
liquefies gelatine, and gives it a
pale wine-red colour.
Agar is coloured a violet tint,
and potato becomes a reddish-
brown.
Urobacillus Duclauxi
(Miquel).—Rods 6 to ‘8 p in
diam., and filaments 2 to 10 win
length. Motile. Spore-formation
present. ;
In gelatine containing ammonia
or urea they develop in the track
of the needle and cause liquefac-
tion. The liquefied gelatine is
viscid.
Broth containing ammonia be-
comes turbid, a sediment forms, and
the liquid gives off an unpleasant
odour.
They occur in sewage.
suppuration in
574
Urobacillus Freudenreichi
(Miquel).—Rods 1 to 1°3 » in width,
and filaments 5 to 6 » in length.
Colonies circular, white.
Inoculated in the depth of gela-
tine growth occurs in the track of
the needle, and a pure white growth
on the surface, followed by slow
liquefaction.
In broth they produce turbidity.
They decompose urea.
They occur in air, sewage and |
dust.
Urobacillus Maddoxi (Miquel). |
—Rods 1 p» in width, 3 to6 » in
length, and involution forms.
Inoculated in the depth of gela-
tine containing urea they produce
white colonies and crystals.
In broth they produce turbidity.
They decompose urea.
They occur in sewage.
Urobacillus Pasteuri (Miquel).
—Rods attaining 1-2 » in width,
and 4 to 6 » in length, singly and
in pairs. Spore-formation present.
They grow in ammoniacal gela-
tine, slowly liquefying it and form-
ing crystals. The liquefied gelatine
is viscid.
They ferment urine, producing
a copious deposit of crystals.
They were isolated from decom-
posing urine.
Urobacillus Schutzenbergi
(Miquel).—Short rods ‘5 » in width,
1 p» in length.
They rapidly liquefy gelatine.
On agar they form a white layer.
They grow readily in broth,
especially after the addition of
urea. The liquid is made cloudy,
but after « few days it becomes
clear again.
They occur in water.
Vibrio rugula (Miiller).—Rods
and threads, 6 to 16 » long, about
‘5 to 2:5 » thick. The rods are
either simply bowed, or possessed
of one shallow spiral (Fig. 230): |
DESCRIPTION OF SPECIES.
They bear a flagellum at each end.
The rods form swarms when caus-
ing decomposition, and then, or
after, grow out into threads, curved
in a screw-like manner. In the
1020.
A. Bowed threads.. B. Slightly-
Fic. 230.—Visrio RucuLa, x
curved rods. C. Rods swollen pre-
Fay to spore-formation. D.
ods swollen at the spore-forming
HE. Various stages of the
(Prazmowski. ),
end.
developing spores.
next stage of development the rods
cease to move, and become swollen
with granular contents. One ex-
tremity develops an enlargement,
giving the rod the appearance of
a pin. The spore formed by
the contraction of the plasma in
the swollen end finally becomes
globular.
The vibrios appear in vegetable
infusions, causing fermentation of
cellulose.
APPENDICES.
575
APPENDIX I.
YEASTS AND MOULDS.
Yeast-fungi and mould-fungi, like bacteria or jission-fungi, are
achlorophyllous Thallophytes. They belong to two separate orders—
the Sacchuromycetes and Hyphomycetes—which are intimately related
to each other, but quite distinct from bacteria. Their germs occur
widely distributed in air, soil and water, and are constantly
encountered in bacteriological investigations. In addition, many
species are of hygienic and pathological interest and importance in
being either accidentally associated with, or the cause of various
morbid processes and fermentations. For a complete account of
all the described species and full details of the various forms of
development, reference must be made to botanical and other
works.* <A description of certain species is appended here, and may
afford some useful information to the worker in a bacteriological
laboratory.
YEAST-FUNGI OR SACCHAROMYCETES.
Saccharomyces cerevisiz (Z’orula cerevisie).—Cells round or
oval, 8 to 9 p long, singly or united in small chains. Spores
occur three or four together in a mother-cell, 4 to 5 mw in diam.
S. cerevisie, S. pastorianus and S. ellipsoideus are active alcoholic
ferments. According to Jérgensen they will produce in fourteen
days in beer-wort from 4 to 6 per cent., by volume, of alcohol.
Saccharomyces ellipsoideus (Hansen). I.—Elliptical cells,
mostly 6 p long, singly or united in little branching chains. Two to:
four spores found in a mother-cell, 3 to 3°5 p in diam. Cultivated
on the surface of wort-gelatine they produce in eleven to fourteen
days, at 25° C.,a@ net-like growth by which they can be recognised
* Sachs, Teat-book of Botany ; Jorgensen, Micro-organisms and Fermen-
tation.
577 37
578 APPENDICES.
with the naked eye. II.—Round, oval, and rarely elongated
cells. They produce yeast-turbidity. There are two so-called
disease-yeasts allied to this species. The colonies of one kind form
a network. This yeast causes turbidity in beer, and a bitter after-
taste. In the other kind the colonies are sharply defined. It
produces a disagreeable aromatic taste to beer, and an astringent
after-taste. It is widely distributed, and is the principal agent in
accidental fermentation. i
Saccharomyces conglomeratus (Reess).—Cells round, 5 to
6 » in diam., united in clusters, consisting of numerous cells
produced by budding from one or a few mother-cells. There are
2 to 4 spores in each mother-cell. They occur on rotting grapes
and'in wine at the commencement of fermentation.
Saccharomyces exiguus (Reess).—Conical or top-shaped
cells, 5 p long, and reaching 2°5 m» in thickness, in slightly
branching colonies. Spore-forming cells are isolated, each contain-
ing 2 or 3 spores in a row. They occur in the after-fermentation
of beer; but, according to Hansen, they do not produce disease
in. beer.
Saccharomyces Jérgensenii (Lasche).—Cells small, round
or oval. On the surface of wort-gelatine the culture is greyish-
white, and the gelatine is slowly liquefied. They ferment saccharose
and dextrose, but not maltose. When grown in wort with other
yeasts they are rapidly crowded out.
Saccharomyces pastorianus, I.—Cells oval or club-shaped.
Colonies consist of primary club-shaped links, 18 to 22 yw long,
which build lateral, secondary, round or oval daughter-cells, 5 to
6 » long. Spores 2 to 4. They occur in the after-fermentation
-of wine, fruit-wines, or fermenting beer, and in the air of breweries.
‘They produce a bitter taste and unpleasant odour and turbidity
in beer. II.—Cells mostly elongated, but also oval or round.
‘Cultivated on the surface of gelatine and yeast-water a growth is
produced with smooth edges, by which it can be differentiated
from No. III. They occur in the air of breweries, but do not
produce disease in beer. III.—They produce yeast-turbidity in
beer. On the surface of yeast-water gelatine the cultures, after
sixteen days, have hairy edges.
Saccharomyces apiculatus.—Cells lemon-shaped, both ends
bluntly pointed, 6 to 8 » long, 2 to 3 4 wide. Budding occurs only
at the pointed ends. Rarely united in colonies. Spores unknown.
They occur with other yeasts in various accidental fermentations
and in ripe fruits. .
YEASTS AND MOULDS. 579
Saccharomyces spheericus.—Cells varying in form; the
basal ones of a colony oblong or cylindrical, 10 to 15 » long,
5 p thick; the others, round, 5 to 6 w in diam. United in ramified
families. Spores unknown.
Saccharomyces anomalus (Hansen).—Cells small, oval, and
sometimes elongated. Spores are hemispherical, with projecting rims
at the base. They were found in impure brewery yeast.
Saccharomyces mycoderma (Mycoderma cerevisice et vini).—
Cells oval, elliptical, or cylindrical, 6 to 7 « long, 2 to 3 yp thick,
united in richly-branching chains. Spore-forming cells may be
20 » long. Spores 1 to 4 in each mother-cell. The colonies in
gelatine are greyish and filmy. They form the so-called “mould”
on fermented liquids, and develop on the surface without exciting
fermentation. When forced to grow submerged, a little alcohol is
produced, but the fungus soon dies. They occur on wine, beer, fruit-
juices and sauerkraut.
Saccharomyces albicans (Oidiwm albicans, Fungus of thrush).
—Cells round, oval, or cylindrical, 3-5 to 5 » thick; the cylindrical
cells 10 to 20 times as long as they are thick. The bud-colonies
mostly consist of rows of cylindrical cells, from the ends of which
oval or round cells shoot out. Spores form singly in roundish cells.
In plate-cultivations the colonies are pure white. In the depth
of gelatine a filament is formed composed of white colonies, some
with ray-like processes extending into the gelatine. On potato the
fungus forms a rapid white growth, and on bread also. They
ean be easily cultivated in a nutrient solution containing sugar
and ammonic tartrate. The cells germinate according to the rich-
ness of the fluid in sugar; they either grow into long threads,
or, in a very strongly saccharine solution, many daughter-cells are
formed and bud out in various directions. According to Klemperer
the thrush-fungus is pathogenic in rabbits, death taking place
twenty-four to ‘forty-eight hours after an intravenous injection of
a pure-culture. Long mycelial threads are found in the internal
organs. They occur on the mucous membrane of the mouth,
especially of infants, in greyish-white patches, which consist of
epithelium, bacteria, yeasts, and the mycelia of various moulds.
Saccharomyces pyriformis (Marshall Ward).—Cells oval.
They convert saccharine solutions containing ginger into ginger-
beer. They occur with other micro-organisms in the so-called
“ ginger-beer plant.”
Saccharomyces glutinis.—Cells round, oval, or short
cylinders, 5 to 11 » long, 4 mw wide, isolated, or united in twos.
580 APPENDICES,
Cell-membrane and contents are. colourless in the fresh state, but
when dried and re-moistened ‘possess a pale-reddish nucleus in the
middle. Spore-formation unknown. They form rose-coloured, slimy
spots on starch paste, and on sterilised potatoes. The colouring
matter is not changed by acids or alkalies.
Saccharomyces ilicis (Grénlund).—Cells spherical. Spore-
formation present without vacuoles. Cultures on the surface of
gelatine have a powdery appearance. They produce about 2°8
per cent., by volume, of alcohol in beer-wort, and cause a disagree-
able, bitter taste. They were obtained from the fruit of Ilex
aquifolium.
Saccharomyces aquifolii (Grénlund)—Cells large and
spherical. Spores contain vacuoles. Cultures on gelatine are
variable, smooth and shining, or powdery. They produce about
-3°7 per cent. alcohol in beer-wort, and cause a sweet taste with
bitter after-taste. They also were obtained from the fruit of Zlex
aquifolium.
Saccharomyces Marxianus (Hansen).—Cells elongated.
They develop a mycelial growth on solid nutrient media. They
occur on grapes.
Saccharomyces membranefaciens (Hansen).—Cells elon-
gated and vacuolated. Spore-formation abundant. Cultivated on
wort-gelatine they produce circular, flattened and wrinkled colonies,
greyish, and sometimes with a reddish tinge. The gelatine is slowly
liquefied. They occur in the slimy secretion of the roots of the
elm, and were also isolated from well-water.
Saccharomyces Hansenii (Zopf).—Cells with small spherical
spores. They set up alcoholic fermentation in solutions containing
sugar. They were found in cotton-seed flour.
Saccharomyces Ludwigii.—Cells irregular in form, oval,
bottle-shaped, lemon-shaped, and elongated, and mycelial filaments..
On wort-gelatine the growth is greyish or yellowish.
Saccharomyces acidi lactici (Grotenfelt).—Cells oval, 2 to
4-35.» in length, and 1:5 to 29 win width. Colonies on nutrient
gelatine are porcelain-white. They coagulate milk.
Saccharomyces minor (Engel).—Cells spherical. Spore-
formation present. They are said to be the most active ferment
in the fermentation of bread.
Saccharomyces rosaceus (Pink Torula).—Cells 9 to 10 p
in diam. They form a coral-pink growth in nutrient gelatine,
nutrient agar-agar, or on sterilised potatoes. They are present.
in the air.
YEASTS AND MOULDS. 581
Saccharomyces niger (Black Torula).—Cells also present
in the air. Cultivated in nutrient gelatine they form a black
crust (Fig. 231).
Mowu.p-Funer on HYPHOMYCETES.
The mould-fungi have been divided into
five orders: Hypodermii, Phycomycetes, Asco-
mycetes, Basidiomycetes and Myxomycetes.
The following species, with the orders to
which they belong, are of especial interest :—-
HYPoDERMII.
Ustilago carbo (Mildew, Smut).—Spores
brown, circular ; episporium smooth ; sporidia,
ovoid cells. The spores or conidia occur as
a black powder in the ears and panicles of
wheat, barley and oats.
Tilletia caries.—Spores round, pale
brown; episporium with reticulated thicken-
ings. In germinating, the sporidia grow
out radially from the end of the promyce-
lium ; these, at their lower part, conjugate oe eee
by a cross branch and separate from the vLA. Pure Cuutiva-
promycelium, and at some point of the pair TION ON Porato.
a hypha grows out, on which abundant
secondary sporidia develop. The latter are long, oval cells, which
can in turn germinate. The fungus occurs in the form of a
stinking powder in grains of wheat, which renders the meal im-
pure, and gives it a disagreeable smell.
Uroceystis occulta.—The spores consist of several cells united
together ; partly, large dark-brown cells in the interior, and out-
side, several flat, semicircular, colourless cells. The promycelium
germinates as in Tilletia, but the cylindrical cells produce a hypha,
without, as a rule, previous conjugation. They occur as a black
powder in rye-straw in long disintegrated stripes, which are at first
greyish. The affected plant produces abortive ears.
Empusa muscze.—A spore or conidium of this fungus
alighting upon the white area of the under surface of the body
of the house-fly germinates into a hypha. The latter, penetrating
the skin, forms toruloid cells, which multiply by germination, and
are disseminated in the blood throughout the body of the fly.
582 APPENDICES.
These cells again grow into hyphe, which penetrate the skin, each
forming a conidium, which is cast off with considerable force. The
parasite is fatal to flies, especially in the autumn. They are often
ohserved attached to the walls or window-panes, surrounded by a
powdery substance, consisting of the extruded conidia.
Empusa radicans.—The spores form long hyphe, which pierce
the transparent skin of the caterpillar of the cabbage white butter-
fly. The terminal cells ramify, and fill the body of the caterpillar
with a network of mycelial filaments. The caterpillars attacked
become restless, then motionless, and death ensues.
Tarichium megaspermum.—tThe spores are black in colour,
and provided with a thickened episporium. They occur at the
sides and ends of mycelial threads, attacking caterpillars (Agrotis
segetum).
PHYCOMYCETES.
Saprolegnia.—Colourless threads, forming dense radiating tufts,
occur on living and dead animal and vegetable matter in fresh
water. The filaments penetrate into the substratum, and branch
more or less in the surrounding water. The cylindrical ends of the
threads are shut off by a septum—forming zoosporangia, or mother-
cells, in the interior of which a number of spherical zoospores
develop. These are set free through an apical opening in the
thread, and after a time coming to rest, give rise to new plants.
In the sexual mode of reproduction a spherical bud, the oogoniuwm,
develops at the end of a mycelial thread; from the thread small
processes or antheridia sprout out laterally towards the oogonium
and blend with its protoplasm. The latter breaks up into a number
of oospores, which clothe themselves with a membrane while still
within the mother-cell, and, eventually being set free, grow into
fresh mycelial filaments. The fungus attacks fish and tritons,
and produces a diseased condition of the skin, which may be
ultimately fatal. In salmon it produces the common “disease of
salmon.”
Peronospora infestans.—The conidia-bearers of this fungus
have as many as five branches, each bearing an egg-shaped
conidium. The contents of the conidia falling off and reaching a
drop of moisture, break up into a number of swarming zoogonidia,
which in turn develop upon plants. Fixing themselves to the
cuticle of the host, they throw a germinating filament into an
epidermal cell; after piercing first its outer wall, and then its inner
YEASTS AND MOULDS. 583.
wall, the filament reaches an intercellular space, where the mycelium
develops. This continues to grow and spread throughout the plant,
In tubers it can hibernate and develop in the young shoots in the
following spring. The fungus appears in the form of brown
patches on the green parts of the plants, especially the leaves.
The attacked parts wither and turn yellow or brown in colour.
If the under surface of a diseased leaf is examined, a corresponding
dark spot may be observed, accompanied with a faint greyish-white
bloom, which covers it. The latter consists of the conidia-bearing
branches.
Pilobolus.—The fruit-hyphe possess spherical receptacles
containing conidia. When ripe the receptacles with their conidia
are detached at their bases, and spring by their elasticity to some
distance. The fungus occurs as glassy tufts on the excrement of
cows, horses, ete. A cultivation can generally be obtained by
keeping fresh horse-dung under a bell-glass.
Mucor mucedo.—Hyphe colourless, simple or branched ; spo-
rangia yellowish-brown or black; spores ovoid. They form the
familiar white mould on fruits, bread, potatoes and excreta, and
penetrate into the interior of nuts and apples. A network of
fibrils develops in the substance of nutrient gelatine, with forma-
tion of sporangia on the free surface. The germination of the
spores and development into hyphe can be observed in a few
hours if the fungus be cultivated in a decoction of horse-dung.
Mucor racemosus.—Hyphe short; sporangia, yellowish to
pale-brown; spores round. By continued cultivation in liquids
saturated with carbonic acid, the hyphze become still shorter
and exhibit a yeast-like sprouting. These yeast-like or toruloid
cells can, when the carbonic acid is withdrawn, germinate into
normal mycelium. They occur on bread and decaying vegetable
matter.
Mucor stolonifer (Lichtheim).—Mycelium grows in the air and’
then bends down and re-enters the nutrient substratum ; sporangia
black, and spores globular. The mycelium can penetrate through
the shell of eggs, and, form conidiophores within them.
Mucor aspergillus (Lichtheim).—Fruit-hyphz thinned at the
base, and with many fork-like divisions ; dark-brown spores.
Mucor phycomyces (Lichtheim),—Mycelium thick-walled ;
olive-green fruit-hyphe ; black sporangia, and oblong spores.
Mucor macrocarpus (Lichtheim).—Spindle-formed, pointed
spores.
Mucor fusiger (Lichtheim).—Ovoid spores.
584 APPENDICES.
Mucor mellittophorus (Lichtheim).—Spores elliptical. Found
in the stomach of bees.
Mucor corymbifer (Lichtheim).—This fungus forms branched
fruit-hyphe. The sporangia have a smooth membrane. It has
been found in the external auditory meatus, and on bread it forms
a dense snow-white growth. Pathogenic in rabbits.
Mucor rhizopodiformis (Lichtheim).—The spores of Mucor
rhizopodiformis and Mucor corymbifer, when introduced into the
vascular system of rabbits, can germinate in the tissues, especially
in the kidneys, where they set up hemorrhagic inflammation.
Dogs are immune, and.only artificial mycosis is known. It occurs
on bread.
Mucor erectus.—Resembles Mucor racemosus. It occurs on
rotting potatoes.
Mucor circinelloides.—Mycelium much branched, and
sporangium carrier is curved.
Mucor spinosus.—Sporangia chocolate. Columella has short
processes or spines.
ASCOMYCETES.
Oidium Tuckeri.—Fruit-hyphe bearing single ovoid conidia.
Observed in the form of brown patches, covered with a white mildew-
like layer on the leaves, branches and young fruit of the vine,
producing “ grape disease.”
Oidium lactis.—Fruit-hyphe simple, erect and colourless,
bearing at their ends a series or chain of conidia. In some cases,
the fruit-hypha branches beneath the chain of spores. Spores are
short cylinders. The conidia germinate into filaments of varying
length, which by subdivision form septate mycelial hyphe; these
and their branches give rise in turn to spores or conidia. The
fungus is deeply stained by the ordinary aniline dyes. In a-plate-
cultivation the colonies appear as white points, and develop into
delicate stellate colonies which ultimately coalesce and form a fine
mycelial network covering the surface of the gelatine. The gela-
tine is not liquefied. The growth on the surface of agar is similar
to that on gelatine. The fungus occurs in sour milk.
Achorion Schonleinii (Fungus of favus).—Threads branching
at right angles, Favus in.man forms yellow crusts on the hairy parts
of the body. The crusts are composed of epidermis and mycelial
filaments and spores. In plate-cultivations whitish colonies are
formed surrounded by liquefied gelatine. Cultivated on the surface
YEASTS AND MOULDS. 585
of gelatine the growth resembles that of Tricophyton tonsurans, but
the liquefaction takes place more slowly, and there is a more fistinct
yellow colour, On agar the growth is white, dry and firmly
adherent.
Tricophyton tonsurans (Fungus of ringworm).—M ycelial
filaments and spores occur on the crusts and in diseased hairs.
In plate-cultivations white colonies are formed, and liquefaction
quickly follows. In test-tube cultivations the gelatine is liquefied
Fic. 232.—Hrap anp Neck or Cate witH AbvANcED Rineworm (Brown).
and the fungus forms a membrane on the liquid jelly which is white
above and yellow beneath. The surface of the growth is powdery
In man the disease varies in appearance in different parts of the
body. Cattle, horses and dogs also suffer from ringworm ; but sheep
and pigs rarely, if ever. The disease is very common in calves.
Sometimes a. small portion of the skin is diseased; in other cases,
the head, neck, chest and abdomen, or even the whole trunk, may be
covered with scabs or crusts. There is often loss of hair in patches,
and the skin may be covered with scurf. The disease is transmissible
586 APPENDICES,
to the human subject. In one case, according to Brown, seven
grooms were infected on the arms from a grey pony which was
suffering from the disease in an aggravated form.
Fungus of fowl-scab.—Fowls are liable to a disease similar
to favus, According to Schiitz this disease is characterised by
greyish-white patches on the comb and wattles of fowls, which may
extend over the neck and body. On nutrient gelatine a white
mycelium is formed; and the gelatine is liquefied, and acquires a
reddish tint. The fungus can be readily cultivated on bread-paste,
agar-agar and potato. Cultures inoculated in fowls produce the
disease, but have no effect on mice and rabbits.
Fungus of mouse-favus.—Mice suffer from a form of favus
which can be communicated to healthy mice by inoculation of scabs
or infected skin (Nicolaier). On nutrient agar the fungus forms a
thick mycelium, at first white, and later of a red or reddish-brown
colour. Mice can be infected with cultures.
Microsporon furfur.—This fungus occurs in Pityriasis
versicolor, Grawitz regarded it as identical with Oidium lactis, and
it is very closely related. Cultivated on gelatine the jelly is hollowed
out and the mycelial growth sinks down, and is yellowish in colour.
‘Oidium albicans.—Vide Saccharomyces albicans.
Aspergillus glaucus (Zurotium aspergillus glawcus).—Mycelium
at first whitish, becoming grey-green or yellow-green. Spores
grey-green, thick-walled. It is found on various substances, chiefly
cooked fruit, and is non-pathogenic.
Aspergillus repens (Hurotium revens, De Bary).—Fruit-heads
fewer than in the above, which are at first pale and then blue-green
to dark-green in colour. Conidia mostly oval, smooth, colourless.
or pale to grey-green,
Aspergillus flavus.—Gold-yellow, greenish and brown tufts.
Fruit-heads round, yellow, olive-green or brown. Conidia round,
seldom oval, sulphur-yellow to brown in colour. Saprophytic in
man, pathogenic in rabbits.
Aspergillus fumigatus.—Greenish, bluish or grey tufts.
Fruit-heads long and conical. Conidia round, and rarely oval,
smooth, mostly pale and colourless. This fungus occurs on bread,
and has been found in the human lungs, external auditory
meatus and middle ear, and in the lungs of birds. The spores
introduced into the vascular system of rabbits, or into the peritoneal
cavity, establish metastatic foci in the kidneys, liver, intestines,
lungs, muscles, and sometimes in the spleen, bones, lymphatic
glands, nervous system and skin.
YEASTS AND MOULDS. 587
Aspergillus niger (Lurotium aspergillus niger, De Bary).—
Dark chocolate-brown tufts. Conidia round, black-brown, or grey-
brown when ripe. This mould can be cultivated readily on bread
moistened with vinegar, on slices of lemon, and on acid fruits and
liquids. It flourishes best of all, according to Raulin, in a liquid
of the following composition :—
Grammes.
Water. : 1500:
Sugar-candy ‘ : : ‘ : 70:
Tartaric acid. 3 : : ‘ i 4:
Nitrate of ammonia . : ‘ : 4:
Phosphate ‘ : : : : ‘6
Carbonate of potassium _ . ‘ ‘ i : 6
5 » Magnesium . : . “4
Sulphate of ammonia é : F : F 25
‘5 », zine : : : ‘ : ‘07
By », iron : : ; ‘ 07
Silicate of potassium 3 , ‘07
It was also found that the fungus grew best when the liquid
was spread out in a layer 2 or 3 cm. in depth in a shallow dish;
and a temperature of 35° C. proved to be the most favourable.
The abstraction of zinc from the nutritive liquid reduced the weight
of a crop from 25 (the average) to 2 grammes, and the presence
of sgas000 part of nitrate of silver, or z54yg part of corrosive
sublimate, stopped the growth altogether. It is saprophytic in the
living body.
Meruop ofr Examinine Aspercitius NIGER.
Species of aspergillus stain intensely with carmine, fuchsine or methyl-
violet ; but to examine Aspergillus niger with a high power a little
special technique is employed, as follows :—A drop of glycerine is placed
on a clean slide, and a drop of alcohol on a cover-glass. With a fine pair
of forceps a few of the fruit-hyphz with their black heads are immersed
in the alcohol. The cover-glass is then turned over on to the drop of
glycerine, and the slide held in the flame of a Bunsen burner till the
spores or conidia are dispersed. To make a permanent preparation
remove the cover-glass, and transfer the fruit-hyphe so treated to a
mixture of glycerine and water (1 to 5); a drop may be conveniently
placed ready on a slide provided with a ring of Canada balsam. The
specimen is then, permanently mounted by employing a circular cover-
glass, and surrounding it with a ring of cement in the usual way.
588 APPENDICES.
Aspergillus ochraceus.—At first flesh-coloured, and then
ochre-yellow heads.
Aspergillus albus.—Pure-white fruit-heads..
Aspergillus clavatus.—Club-shaped fruit-heads on long stems.
Aspergillus nidulans.—Bread and potatoes acquire a reddish-
brown colour. Pathogenic in rabbits. Occurs on bread.
Aspergillus subfuscus.—The growth is olive-yellow in colour.
Pathogenic in rabbits. Occurs on bread.
Aspergillus flavescens.—The growth is yellowish-green.
Pathogenic in dogs and rabbits. Occurs on bread.
Penicillium glaucum.—Occurs as a white, and later a blue:
green, mould, on which dew-like drops of liquid may appear. Its
spores are present in large numbers in the air, and are liable
to contaminate cultivations. The fruit-hypha bears terminally a
number of branched cylindrical cells, from which chains of greenish
conidia are developed. It is the commonest of all moulds,
Botrytis Bassiana._lyphe and spores colourless. Hyphe
usually simple, but sometimes united in arborescent stems. It is
the cause of muscardine, a fatal disease of silkworms, and occurs
also in various other caterpillars and insects.
Chionyphe Carteri.—Mycelial filaments observed by Carter
in Madura disease.
APPENDIX IL.
HAMATOZOA.
HEMATOZOA IN MAN, BIRDS AND TURTLES.—HAMATOZOA IN
EQUINES, CAMELS, RATS AND FISH.—HAMATOZOA IN FROGS.
Hematozoa in Man (Manarta).
Iv 1880 Laveran, in Algiers, noticed the existence of peculiar
structures in the blood of a patient suffering from malaria, and
his researches were communicated to the Academy of Medicine in
Paris in 1881 and 1882, and subsequently published in extenso in a
treatise on the subject.
Laveran described various bodies which he was led to regard as
different stages in the life-history of the same micro-parasite. The
most striking forms were cylindrical elements: with pointed extre-
mities. They were crescent-shaped and pigmented in the middle.
There were other forms, more frequently found, which were either
free in the serum or in contact with the red _blood-corpuscles.
They were more or less spherical, pigmented, and endowed with
ameboid movement. Other forms, again, were provided with motile
filaments three or four times as long as the diameter of a red blood-
corpuscle. And, lastly, there were little masses of hyaline material,
which Laveran regarded as dead forms. :
These observations at first attracted little attention; but they
have since been confirmed and extended by Richard, Councilman and
Abbot, Marchiafava and Celli, Golgi, Sternberg, Osler, the author,
Vandyke Carter, Manson, and others, and their importance fully
recognised.
The different forms assumed by the hematozoon of malaria may
be described in two groups: those within the red blood-corpuscles,
and those free in the serum.
Intra-corpuscular bodies.—These are of three kinds. First,
589
590 APPENDICES.
structureless protoplasmic bodies much smaller than, and within
or attached to, the red blood-corpuscles (Fig. 233). These rapidly
change their shape, exhibiting amcboid movement. They were
first described by Marchiafava and Celli, and possibly represent the
first stage in the life-history of the hematozoon. Marchiafava and
Celli suggested the name Plasmodium malarie. Second, minute
Fic. 233.—Non-PIGMENTED AM@BOID Forms (Marchiafava and Celli).
masses of finely granular or of hyaline protoplasm enclosing granules
of pigment (Fig. 234). These forms are sometimes present in large
numbers, and at other times can be found only with difficulty.
They are more or less spherical, but exhibit amcboid movement,
and rapidly change their form. The pigment granules are also in
active movement. There may be one or more of these amceboid
Fic. 234.—Picmentep AmM@BoIp Forms (Golgi).
bodies to a blood-corpuscle, and they vary in size; one may occupy
the whole of the corpuscle. In cases of pernicious malaria, similar
bodies may be seen, in tissue sections, in the corpuscles filling the
capillaries. Third, forms which appear like isolated grains, and
larger homogeneous bodies surrounded by clear spaces which change
in outline.
Extra-corpuscular bodies.—These are the most striking, and
—
Fic, 235.—SeMi-LUNAR Bopies or LAVERAN (Golgi).
perhaps the most interesting, forms. J irst, the semi-lunar bodies
of Laveran. These are crescent-shaped bodies, sometimes pointed
ANIMAL: MICRO-PARASITES. 591
at the extremities, but more usually rounded off (Fig. 235). They
are not always curved; some, indeed, are almost spherical, and
others sausage-shaped. They are motionless. In many specimens
a delicate line is visible on the concave side of the crescent connect-
ing the extremities. On careful examination this is found to be
o\| Ves eNO) Z af
2 2 5288 Se
Fim 3) a6) Oe
lke a) ©
Fic. 236.—Roserre Forms with SEGMENTATION (Golgi).
the edge of a very delicate membrane. The body is composed of
homogeneous protoplasm. Centrally placed is a collection of pigment
granules, which on careful examination can be distinctly seen to be
in movement. The semi-lunar bodies vary in number in different
cases. Sometimes several can be seen in the field at the same time,
and in other cases they are only observed after a long and patient
search. They are, as a rule, free in the serum; but they have also
been seen within the red blood-cells. Second, finely granular masses
of protoplasm, which arise, according to Golgi, from the intra-
corpuscular pigmented bodies. The pigment is collected in a rosette,
and the protoplasm by segmentation gives rise to a number of small
Fic. 237.—FLAGELLATED Forms (Vandyke Carter).
1. A flagellated spherule; a, the same in the interior of a phagocyte; 0, free
motile filaments.
spherical forms, which are ultimately set free (Fig. 236). Golgi
believes that these changes occur in definite relation to the develop-
ment of the paroxysm. Third, spherical, pear-shaped, or ovoid
bodies, rather smaller than the red blood-corpuseles, and provided
with one or more actively motile flagella (Fig. 237). These flagella
592 APPENDICES.
are long lash-like filaments, which by their activity set the neigh-
bouring blood-corpuscles in motion. Free filaments in active move-
ment have also been observed. owrth, small spherical pigmented.
bodies about one-quarter the size of a red blood-corpuscle, which
exhibit amceboid movement.
Inoculation experiments.—Marchiafava and Celli assert that
inoculation of a healthy subject with blood containing the parasites
will produce a paroxysm of ague with development of the hematozoa.
The pathogenic power of these parasites, however, has not been
established. There has been no cultivation of the parasite outside
the animal body, and reproduction of the disease with a pure culti-
vation. In favour of its being a pathogenic organism, Laveran
points out its invariable presence in some form or other in cases of
malaria; the marked changes it effects in the red blood-cells; the
increase in the number of the parasites in proportion to the severity
of the attack; and, lastly, their disappearance after the admini-
stration of quinine. Others, again, have doubted the parasitic
nature of these bodies, and have looked upon them as representing
pathological changes in the blood-cells.
Laveran first of all suggested the name Oscillaria malarie ; but
subsequently he recognised that these bodies belonged to the animal,
not to the vegetable, kingdom. Osler has suggested that, tempo-
rarily at any rate, the organism should be placed in the genus
Hematomonas of Mitrophanow, thus: “Genus, Hematomonas;
species, Hematomonas malarie. Definition—Body plastic; ovoid
or globose ; no differentiation of protoplasm, which contains pigment.
grains ; flagella variable, from one to four; highly polymorphic,
occurring in (1) amceboid form, (2) crescents, encysted form, (3)
sporocysts, (4) cellular free pigmented bodies.”
EXAMINATION OF THE Hamatozoa oF LAVERAN.
In the Living Condition.—Select a patient by preference who has had
several attacks of malaria, and is markedly anemic. Examine before the
invasion of the febrile paroxysm. Take two perfectly clean cover-glasses
and two clean slides ; wash one of the fingers of the patient with soap
and water, and then cleanse with alcohol ; apply a ligature, and with a
clean needle puncture the thin skin near the root of the nail; touch the
drop of blood which collects, with a clean slide; cover quickly with a
cover-glass, and gently press it if the layer of blood be too thick.
Examine with a 7; 0. 1.
In Stained Preparations—Puncture the finger again if necessary ;
touch the droplet of blood with a clean cover-glass ; apply another cover-
glass; press them gently together, and then slide them apart ; stain with
ANIMAL MICRO-PARASITES, 593
two or three drops of alcoholic solution of methylene-blue ; wash off
excess, and examine in water, or allow the preparation to dry, and mount.
in balsam.
Hematozoa or Brrps.
According to Danilewsky, birds suffer from malaria in both an
acute and a chronic form. The hematozoa are very similar to
those found in malaria in man, and any slight difference may be
attributed to the different character of the blood in birds. Grassi
and Feletti have described two kinds of malarial hematozoa in
birds, one kind belonging to the genus Hemameba and the other
to the genus Laverania.
Hamatozoa oF TuRtLEs,
Danilewsky has also minutely described and figured hematozoa in
the blood of turtles, which in some stages of their life history very
closely resemble those found by Laveran in man.
Hamatozoa or Equives anp Camets (Surra Diszase.)
Surra is a blood disease occurring in horses, mules, and.camels,.
characterised by fever accompanied by jaundice, petechi of mucous
membranes, great prostration, and rapid wasting terminating in
death. The average duration of the disease is about two months.
No organic lesions are found after death, but a parasite exists in
the blood during life. By means of subcutaneous inoculation, and
by the introduction into the stomach of blood containing the
parasite, the disease, according to Evans, can be transmitted to
healthy animals. The importance of this disease may be realised
from the fact that on one occasion in India the 3rd Punjab Cavalry
lost no less than three hundred horses from it.
The disease has not been observed to be contagious or infectious.
in the ordinary sense, but the possibility of its conveyance by means
of large brown flies has been suggested. These flies attack the
horses so vehemently that the blood frequently streams from the
bites ; and the opinion that they propagate the disease is prevalent
among the natives. At the same time it has been particularly
noted that where the disease has broken out the water was very
impure.
Evans discovered a hematozoon, in 1880, in all the diseased horses.
and mules examined ; in all diseased camels, with one exception ; and.
in the dogs which had been subjected to experimental inoculations,
38
594 APPENDICES.
Evans stated that when he first discovered the parasite he
thought it was a spirillum, but very speedily on closer examination
arrived at an opposite opinion.
To him the organism presented the appearance, when fresh and
active, of an apparently round body, tapering in front to form a
neck and terminating in a blunt head. Posteriorly he described a
tapering tail, from which there extended a long slender lash. At
the head end there appeared in one or two cases a circlet of
pseudopods, and as the body slowly died in serum it gave the
appearance of flattening out. After watching very closely all its
changes of form and movements, Evans came to the conclusion that
there existed on either side of the body two fin-like papille, one
near where the neck began and the other close to where the tail
began. In only very few instances he was able to see the four at
once. He suggested that these processes were of the nature of
pseudopods.
The parasite he described as extremely active in its movements,
with an undulatory, eel-like motion, progressing for the most part
head-end foremost, but occasionally moving in the direction of the
lash when tugging at a corpuscle. In fresh blood these organisms
resembled spermatozoa in colour; but their peculiar characteristic
was the power they possessed of attacking and disintegrating the
red corpuscles.
Occasionally two were observed to unite and swim off as one
body ; but the mode of union was a disputed point. Evans thought
that they joined with their respective heads and tails in the same
direction, overlapping each other; but others to whom they were
shown were of opinion that they fastened with their tails in
opposite directions.
The parasites were not always present in the blood, but were
observed to come and go in successive broods. Evans referred the
organism to Lewis for his opinion as to its nature. Lewis arrived
at the conclusion that the parasite was “more nearly related to
that which he found in the blood of rats than to any other”; but
he was of opinion at the time that they did not appear exactly
the same.
Five years later Surra broke out in British Burma, and Steel
was deputed to investigate the outbreak. Steel confirmed the
communicability of the disease to dogs, horses and mules by
ingestion and inoculation, but he considerably supplemented Evans’
views as to the nature of the disease by careful thermometric
observations : these finally led him to regard the disease as a true
ANIMAL MICRO-PARASITES. 595
relapsing fever, closely resembling relapsing fever in man. At the
same time it is worth recording that_until Steel observed the
presence of the parasite described by Evans he regarded the out-
break as malarious in origin, and provisionally termed it gastric
typhoid. In the Burma outbreak, as in the Punjab~ epidemic,
considerable evidence was adduced in favour of regarding the disease
as being due to bad water supply.
Steel succeeded in staining the organism with aniline dyes, but
his description of the parasite in the fresh state differs very
materially from that given by Evans.
Steel failed to recognise the round. body tapering in front, to a
neck. To him the bodies appeared thick in the middle, gradually
diminishing in size in either direction, with a blunt and rigid
extremity at one end. The opposite end he described as tapering
in such a way as to produce a subspiral prolongation, which was
uncurled and lashed about freely like a whip. This tail was
described as slender in relation to the general size of the parasite;
but under the highest power available the presence of a colourless
flagellum could not be detected, nor, he adds, did the movements of
the blood-constituents indicate its existence. |
Steel also failed to see the slightest sign of the two fin-like papille
on each side as described by Evans—an opinion in which he was
supported by Lewis.
These two observers, Evans and Steel, also differed as to whether
the movement could be called spiral. Steel felt convinced that their
movement was as much of that nature at times as can be expected
from organisms with so open a corkscrew shape; while Evans
maintained an opposite view. In the dried and stained specimens
Steel observed that they retained their subspiral form of body and
markedly spiral form of tail.
Steel found that the disease could be communicated to the dog
and to the monkey, and then discussed the resemblance of the parasite
to the spirillum of relapsing fever in man.
From the different appearances presented by the parasite when
in the living state and when dried and stained, Steel thought that
there was probably a still closer resemblance to the living spirillum
than to the dried and stained one, and argued that the figures of
spirilla like corkscrews must be purely imaginary. Steel, it
must be observed, founded these remarks upon figures in text-
books, and not on photographs or on a practical acquaintance
with the spirillum of relapsing fever. One cannot refrain from
pointing out the value of photomicrographs, for they cannot be
596 APPENDICES.
called into question; and had Steel studied photographs of
spirilla he would not have regarded the corkscrew appearance as
imaginary.
Steel found the parasite in all cases, and further observed that
it appeared as the temperature rose and disappeared during the
apyrexial periods.
From all these observations Steel concluded as follows :—That
relapsing fever of mules is an invariably fatal disorder, characterised
by the periodical occurrence of attacks of high fever, during which a
special organism closely resembling the spirillum of relapsing fever
in man is found in the blood. This organism is one-sixth the size of
a red corpuscle in width and three to six times in length. It is
eel-like, and, when dried and stained, presents a thick portion—the
body—and a spiral tail. The latter takes less of the dye than the
former, and commences as a sudden narrowing of the body, termi-
nating by a fine point. This, he insisted, had nothing of the nature
of an infusorian flagellum. The thick portion tapers in either
direction from its centre, and terminates in front abruptly in a rigid
process, with probably some holdfast organ. The sharpness of the
head end varies in different animals. The body portion he described
as spiral, and so closely in general appearances to resemble the
spirillum of relapsing fever that he concluded that the organism
was undoubtedly a spiral bacterium and named it after its discoverer
Spirocheta Evansi. This view, however, would not be accepted by
Evans, who maintained that, whatever it might be, it was not a
member of the family of bacteria.
In the face of these conflicting opinions Evans, in 1885, submitted
to the author preparations of the organism in the blood as well
as material from the lungs and intestines of a camel that had
succumbed to the disease.
On examining a stained preparation the author found that
with a power of 200 diameters a number of the parasites could
be distinguished in the field of the microscope, and with jj, and
qs ©. i, objectives the individual characteristics were clearly brought
out. These were quite sufficient at once to dispel the idea of its being
a spirillum. It was obvious that it was a more highly organized
micro-parasite, presenting very peculiar and distinctive structural
appearances,
The author came to the following conclusions :—
The somewhat tapering central portion, or body, of the parasite
is continuous at one end with a whip-like lash, and at the other end
terminates in an acutely-pointed stiff filament, or spine-like process.
ANIMAL MICRO-PARASITES, 597
Here and there, possibly from injury or want of development, the
spine-like process appears to be blunted or absent. By very careful
focussing on the upper edge of the central portion, the author
discovered the existence—much more markedly in some of the
parasites than in others—of a longitudinal membrane with either
a straight or undulating margin. The membrane is attached along
the body, arising from the base of the rigid filament, and becomes
directly continuous at the opposite end with the flagellum. In some
cases the edge only is deeply stained, giving the appearance of a
thread continuous with the flagellum, so that one might be easily
led to overlook the membrane, and imagine that the flagellum arose
from the opposite end of the body, at the base of the spine-like
process.
Close to the base of the spine-like process a clear unstained spot
is in many parasites easily distinguished; and at the opposite end
there is, in some, the appearance of the deeply-stained protoplasmic
contents having contracted within the faintly-stained membranous
investment. When the longitudinal membrane has a wavy outline
the undulations are much more marked in some cases than in others.
Here and there the wavy outline appears first on the one side of"
the central portion and then on the other; but there never is any
waving outline on both sides of the same part of the body, and this
was explained by a careful examination, which showed that the
somewhat ribbon-like parasite had become doubled on itself. The
discovery of this undulating membrane at once suggested to the
author an explanation of the lateral pseudopodia described by
Evans. If we imagine that we are looking down upon the parasite,
with the edge of ‘the membrane towards us, one can conceive that
the rapid undulations, first on one side and then on another,
might give an image upon the retina which could be construed
as due to the protrusion of lateral pseudopodia. In stained
preparations no trace of the circlet of pseudopods could be
discovered, and the undulating membrane may account for this
appearance also.
Owing to the somewhat curved and twisted shape of the parasite
and the curling of the flagellum in the stained preparations, it
was difficult to make exact measurements; but the average width,
according to whether the membrane was visible or not, varied from
1 to 2 », and the length of the body from 20 to 30. The flagellum
was about the same length as the body.
Here and there in a stained preparation there were the forms
already described by Evans resulting from the fusion of two para-
598 APPENDICES.
sites. But the union obviously took place by the non-flagellated
ends, for the two flagella were frequently turned in the same
direction, so that the fused parasites resembled, as Evans sub-
sequently suggested, a trophy of buffalo horns. Here and there
more than two parasites had united, forming a stellate group; and
in one case the author noticed that the individuals had apparently
united with their non-flagellated ends just overlapping, so that the
unstained spot in one was just situated in a line with the unstained
spot of the other.
In Evans’s Report, Lewis's opinion is given that these parasites
differed slightly, but still were closely allied to certain flagellated
organisms which had been observed by him in rats in India. On
Fic. 238.—‘‘Surra” PARASITES OCCURRING SINGLY AND Fussp.
(From preparations stained with magenta, x 1200. Lent by Dr. Evans.)
referring to his original memoir, it will be found that his description
and woodcut differed very materially from the Surra parasite as
just described, though a microphotograph which Lewis had appended
to the memoir after it was written, indicated a great similarity
to this organism. To the author the organisms appeared not only
closely allied, but, as far as one can judge from figures and descrip-
tions, morphologically identical with the parasites described by
Mitrophanow in the carp, and as a matter of fact, instead of a
mere resemblance, the rat and the Surra parasites, when stained,
are found to be morphologically identical.
ANIMAL MICRO-PARASITES. 599
Hamatozoa or Rats,
In describing these organisms, Lewis remarked that it was.
strange that they had not occupied attention before, and suggested
as an explanation that possibly European rats did not harbour these
parasites. The author examined a few white rats, but without
success, and then proceeded to examine the blood of common brown
rats, trapped from the London sewers, and discovered that these
organisms are to be found in no less than 25 per cent. of apparently
healthy animals. The first question which naturally. arose was
whether these organisms in European rats were identical with those
described by Lewis in Indian rats.
If we refer to the description given by Lewis, we find that he
states that when he first noticed them he thought they were vibrios
or spirilla. The drop of blood
under examination appeared to
uiver with life; and on dilutin
ie blood, motile filaments ‘oad Sa
be seen rushing through the \
serum and tossing the blood- :
corpuscles about in all directions.
The filaments were pale and
translucent, without any trace
of visible structure or granu-
larity, and they were more un-
dulatory in movement than Fic. 239.— Parasites In THE BLoop
spirilla. A corpuscle might be or Rats (Lewis).
observed to quiver, and this
could be distinctly traced to be due to the existence of a flagellum,
apparently a posterior flagellum, as the organisms seemed generally
to move with the thicker end forward; no flagellum could be
detected at the opposite end. The greater number of the figures
in the woodcut (Fig. 239) are described as representing these
organisms a few hours after the blood had been obtained, when
their movements are not so rapid, and the flagellum becomes more
easily recognisable.
This observation led Kent, who named the organism Herpetomonas
Lewisi, to remark that if, as Lewis is inclined to maintain, that
organ “propels instead of draws the animalcule through the in-
habited serum, we have presented a structural and functional feature
without parallel among the other representatives of these Protozow
flagellata, the recognition of which would demand the creation of a
600 APPENDICES.
distinct generic and family group for the reception of these singular
organisms.” In his later paper, however, Lewis came to the con-
clusion that, like the generality of flagellated organisms, the rat
parasites moved with the lash in front.
On careful examination the plasma which constituted the thicker
portion of their substance was observed to suddenly swell out so as
to divide the body into two parts, as seen in the centre of the figure ;
at other times two or three such constrictions or dilatations were
detected, and at other times the body assumed an arrow shape, as
depicted at the lower part of the figure. When dried, and stained
with a little weak solution of aniline-blue, the body presented a very
different appearance. It was found to have contracted irregularly,
and to manifest a somewhat granular and shreddy appearance,
suggestive of a coagulated fibro-albuminous substance. The body
portion became flattened towards its middle to double its original
width, and both ends almost acutely pointed, while the flagellum
was only partly visible. After fixing with osmic acid they measured
0-8 to 1 » in width, and 20 to 30 w in length; the flagellum was about
as long as the body: so that the total length of the organism was
about 50 p. Lewis detected these parasites in 29 per cent. of the
species Mus decumanus and Mus rufescens, but failed to find them
in mice. He considered that they had many features in common
with motile organisms of vegetable origin; but they appeared to
approach much more closely to the Protozoa, more particularly
several of the species of Dujardin’s Cercomonas. He points out
that many, however, believe that these organisms are zoospores and
not animalcules. To him they also seemed to be not unlike the
flagellated parasite described by Biitschli.
The latter observer detected flagellated organisms (Leptomonas
Biitschlii) in the intestinal canal of a free nematode (Trilobus
gracilis). They, too, form stellate colonies, like the Surra parasite,
owing to their being attached by their non-flagellated ends. When
detached from these colonies they presented a somewhat spindle-
shaped body about 11 » in length, with a somewhat thick flagellum
about double this length, so that the total length of the protozoon
would be 33 p, or, as Lewis states, about half the length of the
flagellated organism in the rat's blood. Near the base of the
flagellum, Biitschli’s protozoon presented a contractile vacuole,
but Lewis was unable to detect any such vacuole in the rat
hematozoa.
In conclusion, Lewis observed that very probably these organ-
isms corresponded with the vermicules observed by Goss in the
ANIMAL MICRO-PARASITES. 601
blood of a field mouse, and he also mentions that Chaussat found
minute “‘ nematodes” in the blood of a black rat.
Wittich discovered in the blood of hamsters whip-like bodies with
lively movements. They resembled frog's spermatozoa, possessing
a thick portion continued into a long lash-like thread. Wittich
considered them identical with the organisms described by Lewis, and
they also were observed in apparently healthy animals. Koch later
met with the same organisms.
Like Lewis, the author found that the blood of the common
brown rat in England appeared to quiver with life, and that the
parasites were extremely difficult to examine until their movement
was arrested for a moment or they became imprisoned in the serum
areas. After examining with various powers, from a 4 dry
to a ; 0. i, of Powell and Lealand, the author came to the
following conclusion :—That they are polymorphic, presenting for
Fic. 240.—A Mownap 1n Rav’s Bioop. The organism is represented at partial
rest with its posterior filament impinging on a corpuscle, and showing the
undulating longitudinal membrane, the long flagellum, and the refractive
spherules in the granular protoplasm ( x 3000).
the most part slightly tapering bodies which terminate at one end in
astiff, immotile, acutely-pointed flexible filament or spine-like process,
and at the opposite end are provided with a long flagellum, while,
longitudinally attached, a delicate undulating fin-like membrane can
be traced, which starts from the base of the posterior filament, and
becomes directly continuous with the flagellum (Fig. 240).
With careful illumination the body is found to be distinctly
granular, with one or more highly-refractive spherules. When the
rapid movement is arrested the undulating membrane is distinctly
visible. The best opportunity occurs for seeing this when the
organism comes to partial rest with its stiff filament against a
corpuscle, as if to obtain a point dappui, while lashing its flagellum
in all directions (Fig. 241, 6). At other times, when the parasite has
impinged with its posterior extremity against a corpuscle, or the
stiff filament is apparently entangled in débris, the movements of
the organism give one the idea of its endeavouring to set itself free,
602 APPENDICES.
but the author has not been able to persuade himself that they
attack and disintegrate” the red blood-corpuscles.
In the active state the thicker portion, or body, appears to
twist and bend from side to side with great activity. The organism
can turn completely round with lightning rapidity, so that the
flagellum, at one moment lashing in one direction, is suddenly
observed working in the opposite direction. Then suddenly the
organism makes progression, and it can be distinctly seen to move
in the direction of the flagellum, the flagellum threading vis way
between the corpuscles and drawing the rest of the organism after
it. Currents set up by evaporation may undoubtedly here and
there produce the appearance of the organism “ wriggling along”
with its flagellum posterior; but the author was convinced, after
Fic. 241.—Mownaps 1n Rat’s Bioop, x 1200. a, A monad threading its way among
the blood-corpuscles ; b, another with pendulum movement attached to a cor-
puscle ; c, angular forms ; d, encysted forms ; ¢ and f, the same seen edgeways.
hours of patient observation, that in the normal mode of progression
the flagellum acts as a tractellum and not as a pulsellum. By
treating cover-glass preparations with osmic acid the appearances
obtained are very similar to those shown in Lewis’s photographs,
so that there is no doubt, in spite of the descriptions not completely
according, that they are one and the same organism. There was a
great likeness to the organisms described by Mitrophanow, and to the
Surra parasite ; and when the author had stained the rat parasites,
the closest examination confirmed his belief that they were morpho-
logically identical with the stained parasites of Surra.
Cover-glasses with a thin layer of blood may be passed three
times through the flame of a Bunsen burner in the way commonly
employed for examining micro-organisms, and stained with an
ANIMAL. MICRO-PARASITES. 603
aqueous solution of fuchsin, methyl-violet, or Bismarck-brown,
or with aurantia, nigrosin, and other aniline dyes. The following
method will, however, be found most instructive:—Use freshly
prepared saturated solution of fuchsin or methyl-violet in absolute
alcohol, and put a drop with a pipette on the centre of the prepara-
tion; do not disturb the drop-form for a few moments ; then, before
the alcohol has evaporated, wash off the excess of stain. It will be
found that where the drop rested the organisms will be very deeply
stained, while in the surrounding area the colour will vary in
intensity. By the effect of the different degrees of staining much
Fic. 242,—Monaps 1n Rat’s BLooD STAINED WITH METHYL VIOLET, SHOWING
MEMBRANE UNDER DIFFERENT ASPECTS, BLOOD-CoRPUSCLES, SOME CRE-
NATED AND StaIneD Discs (x 1200).
may be learnt (Fig. 242). In one organism the body and entire
membrane will be equally stained; in another the margin of the
membrane only. In some the posterior stiff filament is stained,
and at its base a darkly stained speck is very striking; and in
other cases, again, the posterior filament is only faintly tinged, or
an unstained spot occurs near its base.
Hamatozoa oF FisH.
In the year 1883 Mitrophanow published a paper in which he
gave an account of organisms in the blood of the mud-fish and the
carp.
In the blood of the mud-fish (Cobitis fossilis) the organisms at
the first glance looked like minute nematodes, but the appearances.
and changes which took place on further examination showed
nothing in common with worms (Fig. 243). As a 1 per cent. salt.
solution had been added to the blood under examination, it occurred
604 APPENDICES.
to Mitrophanow that they were possibly the cytozoa described by
Gaule; but this idea was dismissed by the fact that they were
found in blood to which no salt solution was added. Their size
varied from 30 to 40 » in length and 1 to 14 @ in width. At first
their rapid movements baffled exinuiniion, but as the rapidity
lessened there was the appearance of a curling movement in the
body portion and a swinging movement of the lash. The organism
moved in the direction of the lash, the anterior end of the body
being more pointed than the posterior, and gradually fining off into
the lash. When the body seemed to rest, the lash might be seen to
Fic. 243.—ORGANISMS IN THE BLoop o¥ Mup-¥IsH (Heematomonas cobitis), u, First
variety ; b, second variety; ¢, third variety. d, First variety in a state of
diminished activity. e, The same after treatment with osmic acid. (Mitro-
phanow.)
whip out in all directions. As the movement of the body gradually
diminished? it appeared to have a complicated screw form, the axis
of the screw corresponding to the body to which an undulating
membrane is fastened spirally. This could be distinguished when
the organism was dying, because the body in death contracted, and
the membrane then looked like a spiral addition. Thus the
organism consisted of a body, a spiral membrane, and a flagellum.
With higher magnification the organism appeared to consist of a
refractive, strongly contractile protoplasmic substance, which, when
death occurred, formed a shapeless mass. In the same blood two
other forms were observed: one without a membrane, but having
ANIMAL MICRO-PARASITES. 605
two highly refractive spherules in the protoplasm ; and another with
neither membrane nor flagellum, consisting of very granular proto-
plasm with several refractive spherules, and capable of protruding
processes like pseudopodia.
In the carp (Fig. 244) the parasite is perceptibly larger, and
possesses an undulating membrane fastened along the edge of the
long body. When the body bent first towards one side and then to
the other, a wave-like movement was observable at the free edge of
this membrane.
These parasites were found in all the mud-fish examined except
Fic. 244.—OrRGANISMS IN THE BLooD or THE CARP.
a, b, ¢, Hematomonas carassit ; d, ¢, f, g, h, other organisms in the same blood
(Mitrophanow).
one, and in greater numbers in the hot months. In the carp
they were only found occasionally. Mitrophanow described other
varieties, which he considered were possibly not complete organisms,
but developmental forms. He considered that these organisms were
infusoria between the genera Cercomonas and Trichomonas, with
great similarity to the Trichomonas described in the Lieberkuhn’s
glands of fowls and ducks (Eberth).
On account of their special habitat, Mitrophanow suggested a
new genus—Hematomonas, defining this genus as follows :—Parasites
of normal fish-blood, worm-like, actively moving organisms, with
indistinct differentiation of body parenchyma. Bodies pointed at
606 APPENDICES.
both ends, 30 to 40 » long and 1 to 14 » wide. May possess in
front a flagellum, and on one side an undulating membrane.
Species :—
Hematomonas cobitis.—Body provided with a spiral membrane
and a flagellum at the fore-end. Parenchyma of body homogeneous.
Second variety, body and flagellum only. Movement undulatory,
body containing highly refractive spherules. Third variety, plasma-
like body, without membrane or flagellum; quickly changes form
by sending out processes laterally, and contains two to four refractive
spherules. Blood of mud-fish.
Hematomonas carassiiimLong bodies, with narrow membrane
attached along the whole length; less actively motile. Several
forms also observed strikingly smaller than the above; many dise-
shaped. Often seen attached to a red corpuscle, setting them in
motion by their movements. Blood of carp.
The morphological identity of the rat and Surra parasites has been
established by the author, and both szem morphologically identical
with the organism of Mitrophanow. If we follow Mitrophanow, we
must obviously enlarge his genus of Hematomonas. The author does
not agree with Mitrophanow in the advisability of adopting this
entirely new generic name. Mitrophanow suggested this new term
because of the special habitat, normal fish-blood, of the species he
discovered. But the characteristic features of these organisms are the
characteristic marks of the genus Trichomonas. It seems, therefore,
that they are embraced by the old genus Trichomonas, and that there
is no need to create a new one—Hzmatomonas. The common habitat
of these species may be expressed by grouping them together in one
sub-genus—Trichomonas sanguinis ; but the question arises whether
they are distinct species. If it were not for the different description '
given by Mitrophanow of the organism in the mud-fish, the author
would be inclined to say that all these organisms belonged to one and
the same species, which might well be named Trichomonas sanguinis.
The monad in the rat and the-Surra parasite are morphologically
identical with each other, and both, as far as one can judge from
the description, morphologically identical with the monad in the
blood of the carp. We have, however, seen that the organism in
Surra is believed to be pathogenic, and too much stress must not be
laid on morphological identity. There is strong evidence in favour
of believing in its pathogenic properties; but at the same time it
must be borne in mind that the organism has never been isolated
apart from the blood, and the disease then produced by its introduc-
tion into healthy animals. It is quite possible that the parasites in
ANIMAL MICRO-PARASITES, 607
Surra are only associated with the disease, the impoverished blood
affording a suitable nidus for their development, while the con-
taminated water may be the common source of the organism and of
the disease. On the other hand, the organism in the rat is found in
apparently perfectly healthy, well-nourished animals, The author
suggests that the parasites observed in the rat and hamster should
be named after Lewis, Trichomonas Lewisi; the organism in the
mule, camel and horse after its discoverer, Trichomonas ELvansi ;
and that the names Trichomonas cobitis and Trichomonas carassii
should be substituted for the names of the Species described by
Mitrophanow. Thus we should have added provisionally to the
Genus—TRICHOMONAS.
Sub-genus—Trichomonas sanguinis. Definition: Elongated
tapering bodies, provided with a spiral (7. cobitis), or
longitudinal (7. carassii, Lewisi, Hvansi) membrane, ter-
minating in a rigid filament and an anterior flagellum.
Highly polymorphic. Habitat, the blood.
Species.—Trichomonas cobitis (Hamatomonas cobitis
Mitrophanow)—Mud-fish.
Trichomonas carassii (Hematomonas carassii
Mitrophanow)—Carp.
Trichomonas Lewisi (Herpetomonas Lewisi
Kent)—Rat, hamster.
Trichomonas Evansi—(Spirocheta Evansi
Steel)—Horse, mule, camel ; (pathogenic ?).
Hamatozoa OF THE FRoG.
Lankester described an organism which he had discovered in the
blood of the frog (Rana esculenta). It consisted of a minute pyri-
form sac, with the narrower end bent round on itself somewhat
spirally, and the broader end spread out into a thin membrane,
which exhibited four or five folds and was prolonged on one side into
a very long flagellum. The wall of the sac was striated, nucleated
and granular; the membrane undulated during life, and the
flagellum was also motile. It was named Undulina ranarum, but
subsequently recognised as idnetical with Trypanosoma sanguinis
described by Gruby. In the same blood Lankester also discovered
little oblong bodies, in many cases attached to the end of the red
corpuscles, and suggested a genetical connection with the Undulina.
608 APPENDICES.
One or more motionless filaments were occasionally observed attached
to these bodies, Gaule subsequently observed the same bodies, and
regarded them as resulting from the metamorphosis of the cells of
the frog’s blood. Gaule’s observations were refuted by Lankester in
1882, the parasitic nature insisted upon, and the organism named
Drepanidium ranarum. Lankester suggested that they were
probably the young stage of a sporozoon allied to Sarcocystis or to
Coccidium
APPENDIX III.
PSOROSPERMS OR COCCIDIA—AMCBA COLI,
PsoROSPERMS OR Coccrpta,
GREYISH-WHITE nodules may occasionally be found in the liver of a
rabbit, the result of a disease which may be mistaken for tuber-
culosis, This disease often proves fatal, and may occur in an
epidemic form in rabbit warrens. The nodules have cheesy or
purulent contents, which are found, 9n microscopical examination,
to contain great quantities of Coccidium oviforme.
The coccidia pass from the intestine into the bile-ducts. The
walls of the bile-ducts become dilated and folded; and irregular
cavities result from the partial or complete disappearance of the
dividing walls of the altered ducts. The folds are composed of
connective tissues lined with columnar epithelium, and the coccidia,
in different stages of development, are found between the cells, and
free in the cavities of the nodules.
The individual coccidia are egg-shaped bodies. They possess a
thick smooth shell, with an opening, or micropyle, at one end, and
protoplasmic contents which may completely fill the capsule or be
collected into a spherical mass.
After passing from the liver and intestine, these oval bodies
undergo a further development. According to Leuckart, who has
very fully described this parasite, the protoplasmic contents divide
into four masses, and from each is developed a. C-shaped hyaline
rod, the cavity of which is occupied by closely packed granules. In
this condition they remain until they gain access to a fresh host.
Coccidium oviforme has been found in the human liver, and also
in sheep, dogs, and cats. Similar, but not identical, bodies occur in
mice, and also in fish and other cold-blooded animals.
Miescher’s tubes are peculiar structures found in swine, cattle,
sheep, deer, and mice, They consist of a firm envelope inclosing a
number of reniform or bean-shaped bodies.
609 39
610 APPENDICES.
Pfeiffer’s bodies.—Pfeiffer has described certain appearances
which he attributes to coccidia, in epithelial cells in small-pox,
vaccinia, and other vesicular diseases. They are probably only
derived from the cell nucleus, and are not parasites.
Cancer bodies.—In sections of malignant growths stained by
aniline dyes, certain bodies have been found and minutely described
and figured by various investigators, and a causal relation sug-
gested. Darier first described bodies like cysts, with spores, in
Paget’s disease of the nipple. Wickham found similar structures
and figured them. Nils Sjébring described a cancer parasite, and
illustrated his researches with plates. Russell drew attention to
certain bodies in cancerous tumours, with a great affinity for
fuchsine. Soudakewitch, Podwyssozki, Sawtschenko, Ruffer, and
Walker have, among others, contributed to the literature of the so-
called cancer parasites. These bodies appear in the form of refractile
spherical elements, which stain well with reagents, such as the
Ehrlich-Biondi stain. Sections are left in this stain for twenty-four
hours, washed in alcohol, cleared in xylol, and mounted in xylol
balsam. The spherical bodies have sometimes a radiate appearance,
These bodies have not been cultivated, and inoculation experiments
with cancerous tissue have been negative. The opinion is now very
generally held that these bodies are not parasites, but that changes
occur in the cells and nuclei, resulting in the formation of peculiar
structures, which have been brought to light by the use of aniline
dyes and complex staining methods. We are justified in concluding
that the cause of cancer is unknown.
Ballance and Shattock have made repeated attempts to cultivate
parasitic protozoa from malignant tumours, and they have extended
their researches to vaccinia and molluscwum contagiosum, but with
negative results. Sand and water were used as the medium for
these experiments, Cultivations were made from nine scirrhous
carcinomata of the breast, five sarcomata from different sources, two
melanotic sarcomata from horses, and a sarcoma from a dog. In
every instance the result was negative. No traces of protozoic life
could .be found, in spite of examinations at regular intervals, and
repeated for periods of many months. ,
Am@BA Cott.
Losch, Grassi, Kartulis, and others have described an amoeba in
the intestines of patients suffering from dysentery. Lésch adminis-
tered the fresh dejecta of a patient containing the amcbe to dogs,
ANIMAL MICRO-PARASITES. 611
and in one case a mucous mass was passed containing a number
of amebe. Highteen days afterwards the dog was killed, and the
mucous membrane of the intestine was reddened, swollen, and
Fic. 245.—Ama@pa Coit in Intestinan Mucus (Loscu).
ulcerated in three places. The mucus in the rectum and in the
ulcers contained numerous amcebe. Cunningham, who has found
the amcebe in choleraic and other cases, and in the intestine of the
cow and horse, does not attach any importance to their presence.
APPENDIX IV.
APPARATUS, MATERIAL, AND REAGENTS EMPLOYED
IN A BACTERIOLOGICAL LABORATORY.
(A) HisroLogicaL APPARATUS.
Microscope.—For the investigation of micro-organisms a good
microscope with oil-immersion system and a condenser, such as
Abbé’s, is essential, Such instruments are supplied by Zeiss, Leitz,
Reichert & Hartnack in Germany, and Powell & Lealand, Swift
& Baker in England. Zeiss supplies a micrometer eyepiece, with
directions for use. Some such arrangement is essential for the
measurement of bacteria. Other accessories to the microscope are:
A large bell-glass, for covering the microscope when not in use.
About a foot square of blackened plate-glass,
A white porcelain slab of the same size.
Glass bottles, with ground-glass stoppers, for alcoholic solutions of
aniline dyes, etc.
Glass bottles, with funnels, for aqueous solutions of the dyes, and
others provided with pipettes.
A small rod-stoppered bottle of cedar oil, This is recommended by
Zeiss in preference to other oils for his immersion lenses,
Set of small glass dishes or capsules and watch-glasses, for section-
staining, etc.
Stock of best glass slides, in packets of fifty.
Several boxes of round and square thin cover-glasses, in various sizes,
-of the best quality.
Needle-holders, with a couple of platinum needles, and a packet of
ordinary sewing-needles.
Glass rods drawn out to a fine point ; useful for manipulating sections
when acids are employed.
Platinum or plated copper section-lifters.
One pair of small brass or spring-steel platinum-pointed forceps, for
holding cover-glasses.
One pair of brass tongs.
612
APPARATUS, MATERIAL, AND REAGENTS. 613
Collapsible tubes, for containing Canada balsain ; very serviceable for
transport and general use.
Turn-table for sealing cover-glass preparations, with rings of cement.
Boxes for preparations, book-form.
Tickets and labels, various sizes.
Soft rags or old pocket-handkerchiefs, for removing cedar oil from
immersion lens, cleaning cover-glasses, etc.
Chamois leather for wiping lenses.
Warm Stages.—In addition to those already described, Schafer
and Stricker have constructed warm stages for accurate observations.
Schafer’s apparatus consists of a vessel (/f), filled with water which
has been boiled to expel the air, and heated by means of a gas-flame
at g. The warmed water ascends the indiarubber tube (c) to the
PY
wey >
"<<
“ing
Fic. 246.—Scuarer’s WARM STAGE.
brass box (a). The box is pierced by a tubular aperture to admit
light to the object, and has an exit tube (c'), by which the cooled
water from the stage returns to be reheated by the flame g. Atd
is a gas-regulator, so that a constant temperature at any desired
point can be maintained.
Stricker’s stage, in which warm water or steam can be used for
heating, and by the employment of iced water also used for observing
the effects of cold, is shown in Fig. 247. It consists of a hollow
rectangular box, with a central opening (C) permitting the passage
of light. The water makes its exit and entrance at the side tubes
(a, a), and the temperature is indicated by a thermometer in front.
614 APPENDICES.
A more complicated apparatus, combining both a warm stage
and a gas chamber, is shown in Fig. 248. This consists of a rect-
angular piece of ebonite (# #) fixed to a brass plate which rests
on the stage of the microscope. On the upper surface of the ebonite
is‘another brass plate (P), with an aperture (C’/) leading into a brass
.
C a”
1 T 1 T
Fig. 247.—STRICKER’s Warm STAGE,
tube closed below by a piece of glass. To heat the apparatus the
copper wire B is placed on the tube a, and its extremity heated by
the flame of the lamp. The nearer the lamp to the stage the higher
the temperature, which is indicated by the thermometer (¢). To
wv
o
alo z
Fic. 248.—Stricker’s ComBinED Gas CHAMBER AND WaRM Stace.
employ it as a gas chamber the wire B is laid aside, and the gas
is conducted into the chamber by the tube a’, and escapes by the
tube a.
Microtome.—Schanze’s is much in favour in Germany, but
Jung’s of Heidelberg, though a somewhat cumbrous instrument, is
APPARATUS, MATERIAL, AND REAGENTS. 615
preferred by many workers. Smaller accessories, which should be
within reach, are—
A small can of sewing-machine oil.
A soft rag and chamois leather, for wiping the knives immediately
after use.
Stone and leather, for setting and sharpening the same.
Two or three camel’s-hair brushes.
A freezing microtome is very useful: such as Swift’s, which is
used by the author; and the method of embedding in celloidin is
combined with the ordinary process of freezing.
(B) Reacents anp Marerta, Empioyep IN THE PRocEssEs oF
Harpenine, Decatciryinc, EmBrppine, Fixina anp Currine
oF TissvEs.
Alcohol, absolute.
Bergamot oil.
Celloidin.
‘Dissolved in equal parts of ether and alcohol.
Cork, or stock of ready-cut corks.
Ebnevr’s solution. A mixture in the following proportions :—
Hydrochloric acid . : ‘ : : ‘ 5
Alcohol : : 3 3 ‘ é z : 100
Distilled water. 2 5 . 20
Chloride of sodium : : ; ; : 5
Formalin.
Gelatine.
Melted in a small porcelain capsule, and set aside ready to be
re-melted when required for use.
Glycerine gelatine (Klebs).
Best well-washed gelatine. 10
Add distilled water, allow aointiae: to aril up, pour
off excess of water, melt gelatine with gentle heat,
add
Glycerine
ee a few dine of phensl for praca:
10
Gum.
616 APPENDICES.
Kleinenberg’s solution.
Saturated watery solution of picric acid ‘ . 100
Strong sulphuric acid . ‘ : 2
Filter, and add
Distilled water i : ; . 3800
Miiller’s fluid.
Bichromate of potash
Sulphate of sodium : : ; ' : 1
Distilled water : : ‘ : . 100
Osmice acid.
Distilled water ‘ : : . 100
Osmic acid . ; ‘ : ; : : D
Paper trays (or small glass capsules).
Paraffine.
Spermaceti.
Xylol.
(C) Reacents ror Examining anp Srainina MicroscopicaL
PREPARATIONS.
. Acetic acid, strong.
. Alcohol, absolute.
. Alcohol, 60 per cent.
. Aleohol, acidulated. ;
Alcohol. : ; : : ‘ , 100
Hydrochloric acid. ; ‘ 1
em Oo bb Fe
5. Alum Carmine (Grenacher).
Carmine . . : 5 1
Five per cent. soliton of — : 100
Boil twenty minutes; filter when veld,
6. Ammonia, strong.
7. Aniline.
8. Aniline water.
Distilled water j j ; ; . 100
Aniline . i ‘ ; ; ; : 5
Shake well, and filter emulsion.
APPARATUS, MATERIAL, AND REAGENTS, 617
9. Bismarck-brown.
(a) Concentrated solution in equal parts of glycerine and water.
(6) Aqueous solution.
Bismarck-brown ‘ : ; 2
Alcohol . 3 : ; : 3 . 15
Distilled water ‘ B : : : . 85
10. Borax-carmine (Grenacher).
Borax , ‘ : : : 2
Carmine . : 3 3 . : : F 5
Distilled water : ‘ é : : . 100
To the dark purple solution add a 5 per cent, solution of acetic
acid until a red colour is produced ; set aside twenty-four hours ;
filter, and add a drop of carbolic acid. :
11. Cedar oil.
12, Ehrlich-Biondi solution (Heidenhain).
To
Saturated aqueous solution of Orange. G. . 100
Add
Saturated aqueous solution of Rubin. 8. . 20
Pr 5 “i Methyl-green. OO. 50
To the mixture : ‘ : : 5 j 1
Add water. : F . 100
13. Eosin.
(a) Saturated alcoholic solution.
(6) Aqueous solution.
Distilled water. . 100
Eosin. ‘ ‘ ; : 5
14, Ether.
15. Fuchsine.
(a) Saturated alcoholic solution.
(6) Aqueous solution.
Fuchsine . : : : 2
Alcohol . : ‘ : ‘ .
Water . : : . 85
16, Gentian-violet.
(a) Saturated alcoholic solution.
6) Aqueous solution.
(6) Aq sh
Gentian-violet . z :
Distilled water ' : . 100
618 APPENDICES.
17. Gibbes’ solution, for double staining.
Take of
Rosaniline hydrochlorate .
Methylene-blue
Triturate in a glass mortar.
Dissolve aniline oil .
In rectified spirit
and add slowly to the on
Lastly, slowly add distilled water
Keep in stoppered bottle.
18. Giycerine, pure.
19. Hematoxylin solution.
” -Hematoxylin .
Alcohol .
Distilled water
Glycerine
Alum
20. Iodine solution.
- Iodine, pure
Iodide of potassium .
Distilled water
21. Iodine solution (Gram).
Iodine ;
Todide of potassium .
Distilled water
22. Lithium-carmine solution (Orth).
Saturated solution of carbonate of lithium
Carmine .
23. Magenta solution (Gibbes).
Magenta .
Aniline oil
Alcohol (sp. gr. 830)
Distilled water
24. Methylene-blue.
(a) Concentrated alcoholic solution.
(6) Aqueous solution.
Methylene-blue
Alcohol .
Water
15
85
APPARATUS, MATERIAL, AND REAGENTS,
(c) Koch’s solution.
Concentrated alcoholicsolution of methylene-blue
Ten per cent. potash solution . ;
Distilled water
(d) Liffler’s solution.
Concentrated alcoholic solution of methylene-blue
Solution of potash, 1 to 10,000
25. Methyl-violet.
(a) Concentrated alcoholic solution.
(6) Aqueous solution.
Methyl-violet .
Distilled water
(ce) Koch’s solution.
Aniline water .
Alcoholic solution of methyl: valele
Absolute alcohol
26. Neelsen’s solution.
Dissolve fuchsine
In alcohol
Add a. 5 per cent. watery solution of catbalts acid
27. Nitric acid, pure.
28. Orseille (Wedl).
Dissolve pure ammonia-free orseille in
Absolute alcohol
Acetic acid
Distilled water ; :
until a dark red liquid seal Filter.
29. Picric acid.
(a) Concentrated alcoholic solution.
{2) Saturated aqueous solution.
30. Picro-carmine (Ranvier).
Carmine .
Distilled water
Solution of ammonia .
Triturate ; add cold saturated aliases af. picric
acid
100
2:25
100
100
10
100
20
40
619
620 : APPENDICES.
31. Picro-lithium-carmine (Orth).
To above-mentioned lithium-carmine solution
add saturated solution of picric acid. . 23
32, Potash solution.
(a) 1 to 3 per cent.
(2) 10, »
(c) G3 se yg
33. Safranine.
(a) Concentrated alcoholic solution.
(6) Watery solution. : : - 1 per cent. —
34, Sulphuric acid, pure.
35. Salt solution ‘ ‘ : ‘ . 0°8 per cent.
36. Turpentine.
37. Vesuvin.
(a) Concentrated alcoholic solution.
(6) Watery solution.
Water, distilled.
Water, sterilised.
Distilled water can be kept for use in a wash bottle, or far
better in a siphon apparatus. Sterilised water is convenient in
plugged sterile test-tubes, which may be kept close at hand in a
beaker, or tumbler, with a pad of cotton wool at the bottom. The
numbered reagents can be conveniently arranged on shelves within
easy reach. Alcoholic solutions of the aniline dyes and other special
preparations should be kept in bottles with ground-glass stoppers.
Aqueous solutions of the dyes may be kept in bottles with funnel
filters, and the solution filtered before use. To both aqueous and
alcoholic solutions a few drops of phenol, or a crystal of thymol,
should be added as a preservative. For the rapid staining of cover-
glass preparations, it is convenient also to have the most frequently
used stains (fuchsine, methyl-violet) in bottles provided with pipette
stoppers.
(D) Reagents ror Movuntine anp Preservina PREPARATIONS.
Acetate of potash.
Concentrated solution.
Asphalte lac.
APPARATUS, MATERIAL, AND REAGENTS, 621
Canada balsam.
Dissolved in xylol.
Glycerine gum (Farrant’s solution),
Glycerine.
Water.
Saturated solution of arsenious acid,
Equal parts ; mix, and add of picked gum arabic half a part.
Hollis’ glue.
Zine-white.
(EZ) Drawine anp PHorograPHic APPARATUS.
Camera Lucida.—The camera lucida of Zeiss is an excellent
instrument, though many prefer the pattern made by Nachet of
Paris. Combined with the use of a micromillimeter objective, it
affords also a simple method for the measurement of bacteria.
For drawing microscopical appearances, and for illustrating
microscopical specimens with or without the use of a camera lucida,
the following materials should be within reach :—
Pencils.
Hiching pens.
Prepared Indian ink.
Water-colour paints and brushes. .
Ordinary and tinted drawing paper and other usual accessories.
Photo-micrographic Apparatus.—Zeiss of Jena, Seibert &
Kraft of Wetzlar, Nachet of Paris, and Swift & Son of London, may
all be recommended for constructing an arrangement in which the
photographic camera is combined with the microscope.
The best models have been described fully in the chapter on
Photography of Bacteria. The accompanying figure (Fig. 249)
illustrates a model in which the microscope is used in the vertical
position.
For illumination either sunlight or artificial light may be em-
ployed. In the case of sunlight a heliostat is necessary to procure
the best results; but as sunlight is not always available by day, and
it is also more convenient for many to work at night, it is better to
have recourse altogether to artificial light. Excellent results may
be obtained with an ordinary paraffine lamp, or with magnesium,
oxycalcium, or electric light.
622 . APPENDICES.
a
Fic. 249.—VeErticaL Micro-PHoToGRAPHIC APPARATUS.
(F) Sreritisarion APPARATUS.
Steam-steriliser.—A cylindrical vessel of tin about half a metre
or more in height, jacketed with thick felt, and provided with a
conical cap or lid (Fig. 250). The lid is also covered with felt, has
handles on either side, and is perforated at the apex, to receive a
thermometer. Inside the vessel is an iron grating or diaphragm
about two-thirds the way down, which divides the interior into
two chambers—the upper or ‘“ steam-chamber,” and the lower or
““water-chamber.” A gauge outside marks the level of the water
in the lower chamber; this should be kept about two-thirds full.
APPARATUS, MATERIAL, AND REAGENTS. 623
The apparatus stands upon three legs, and is heated from below with
Fic. 250.—Kocu’s Steam-
STERILISER.
dicated by a thermometer inserted through
a hole in the roof; in a second opening a
gas regulator can be fixed. Test-tubes,
flasks, funnels, cotton
sterilised by exposure to a temperature
of 150° C. for an hour or more.
two or three Bunsen burners, or a Fletcher’s
burner. It is employed for sterilising
nutrient media in tubes or flasks, for cooking
potatoes, or hastening the filtration of agar-
agar, When the thermometer indicates
100° C. the lid is removed, and test-tubes
are lowered in a wire basket by means of
a hook and string, and the lid quickly re-
placed. Potatoes or small flasks are lowered
into the cylinder in a tin receiver with a
perforated bottom, which rests upon the
grating and admits of its contents being
exposed to the steam. A larger model is
shown in Fig. 33.
Hot-air Steriliser.—A cubical chest of
sheet iron with double walls, supported on
four legs ; it may also be suspended on the
wall of the laboratory, with a sheet of
asbestos intervening (Figs. 251 and 252).
It is heated with a rose gas-burner from
below, and the temperature of the interior in-
wool, etc., may be
Fic. 252.—Section or Hor-
Fic. 251.—Hor-arr STERILISER. AIR STERILISER.
624 APPENDICES.
(G) Apparatus AND MArerIAL ror PREPARING AND SroRING
Nourrient GELATINE AND NutTRIENT AGAR-AGAR.
Water-bath.— A water-bath on tripod stand is required for
boiling the ingredients of nutrient jellies and for general purposes.
The lid may be conveniently composed of a series of concentric
rings, so that the mouth of the vessel may be graduated to any
size required.
Test-tube Water-bath.—This consists of a circular rack for
test-tubes within a water-bath. It is sometimes employed instead
of the steam cylinder for sterilising nutrient jelly in tubes, by
boiling for an hour for three successive days.
Hot-water Filter.—A copper
funnel with double walls, the inter-
space between which is filled with
hot water. A glass funnel fits in-
side the copper cone, the stem of
the glass funnel passing through
and being tightly gripped by a per-
forated caoutchouc plug, which fits
in the opening at the apex of the
cone. The water in the cone is
heated by applying the flame of a
burner to a tubular prolongation
of the water-chamber. In a more
recent model, as represented in
Fig. 31, this prolongation is dis-
pensed with, and the temperature is
maintained by means of a circular
burner which acts at the same time
as a funnel ring. In Rohrbeck’s
model the funnel of the filter is
connected with a flask, from which
the test-tubes can be easily filled with the liquid jelly (Fig. 253).
Glass Vessels.—A number of glass vessels should be kept in
stock according to requirements.
Fic. 253.—Hort-watTer FILrerine
APPARATUS WITH RING BURNER.
Bohemian hard glass flasks are employed in several sizes, for
boiling nutrient media. The conical forms are especially used in the
larger sizes for storing nutrient jelly.
Glass funnels, large and small, are necessary, not only in the
processes of preparing nutrient jelly, but for filtering solutions of
aniline dyes and for general purposes.
APPARATUS, MATERIAL, AND REAGENTS. 625
A liberal supply of test-tubes should always be kept in stock, as
they are not only employed for the tube-cultivations, but can be
conveniently used for storing bouillon, sterilised water, etc.
Cylindrical glasses graduated in cubic centimetres, 10 ccm., 100
cem., 500 ccm., are required for measuring the liquid ingredients
of nutrient jelly, and also in preparing the various staining
solutions.
A large wide-mouthed glass jar, with a glass cover, is extremely
useful. It must be padded at the bottom with cotton wool for
containing a stock of tubes of sterilised nutrient jelly, and should
be placed within reach on the working table.
Balance and Weights.—A balance, with large pans and set of
gramme weights, is constantly required.
Cotton Wool.—The best or “ medicated” cotton wool should be
procured.
Gelatine.—The gelatine for bacteriological purposes must be of
the very best quality (gold label).
Agar-agar.—This is also called Japanese Isinglass; it consists
of the shrivelled filaments of certain Alge (Gracilaria lichenoides
and Gigartina speciosa).
Peptonum Siccum.
Table Salt.—Prepared table salt can be obtained in. tins or
packets.
Litmus Papers.—Blue or red litmus paper in cheque-books, for
testing the gelatine mixture, etc.
Carbonate of Soda.—aA bottle, containing a saturated solution
of carbonate of soda, and provided with a pipette stopper, may be
kept, especially for use in the preparation of nutrient jelly.
Lactic Acid.
Filter Paper.—For filtering gelatine, stout Swedish filter paper
of the best quality is recommended.
Flannel or Frieze.—This is employed as a substitute for, or
combined with, filter paper in the preparation of nutrient agar-
agar.
(H) Apparatus FoR EMPLOYMENT OF Nourrienr Jeviy in Test-TUBE
AND PLATE-CULTIVATIONS.
Wire Cages.—These cages or crates are used for containing
test-tubes, especially when they are to be sterilised in the hot-air
steriliser ; or for lowering tubes of nutrient jelly into the steam-
steriliser, etc. (Fig. 254). ;
0
626 APPENDICES.
Test-tube Stands.—The ordinary wooden pattern, or the
metallic folding stands, are called into use
for holding cultivations. Pegged racks are
also recommended for draining test-tubes
after washing.
Caoutchoue Caps.—These are caps for
fitting over the cotton-wool plugs, and may
be used in different sizes for test-tubes and
stock-flasks.
Platinum Needles.—A platinum
needle for inoculating nutrient media, ex-
Fic. 254.—Wire Cace amining cultivations, etc., consists of two or
YOR Tes! TUBES: three inches of platinum wire fixed to the
end of a glass rod. Several of these needles should be made with
platinum wire of various thicknesses. A piece of glass rod, about
seven inches long, is heated at the extreme point in the flame of
Fic. 255.—Piatinum NEEDLES; STRAIGHT, HooKED, Looprep.
a Bunsen burner, and a piece of platinum wire, held near one
extremity with forceps, is then fused into the end of the rod.
Some needles should be perfectly straight, and kept especially for
Fic. 256.—Dame CHAMBER FOR PLATE-CULTIVATIONS.
inoculating test-tubes of nutrient jelly. For other purposes the
needles may be bent at the extremity into a small hook, and
others provided with a loop (Fig. 255).
APPARATUS, MATERIAL, AND REAGENTS. 627
Tripod Levelling-stand.—-A triangular wooden frame sup-
ported upon three screw-feet which enable it to be raised or lowered
to adjust the level.
i
wn
Fic. 257.—APPARATUS EMPLOYED FOR PLATE-CULTIVATIONS.
Tripod Stand ; Glass Dish, filled with cold or iced water; Sheet of Plate-glass ;
Spirit Level, and Glass Bell.
Large Glass Plate.—A piece of plate-glass, or a pane of
ordinary window-glass, about a foot square.
Spirit Level.
Glass Bells and Dishes.—Shallow glass bells and dishes, for
making a dozen or more damp chambers im
(Fig. 256), and for completing the apparatus \
for pouring out liquefied nutrient al on i = vA
glass plates or slides (Fig. 257).
Iron Box.—A box of sheet-iron
(Fig. 258), for containing glass plates during
their sterilisation in the hot-air steriliser,
and for storing them until required for use.
Glass Plates.—Small panes of glass,
about six inches by four. Not less than
Fic. 258.—Box For
three dozen are required for a dozen damp Guass Pratss.
chambers.
Glass Benches.—These are necessary for arranging the glass
plates or slides in tiers in the damp chambers (Fig. 256). Metal
Fic. 259.—G Lass BENcHES FOR GLASS PLATES OR SLIDES.
shelves may be substituted for them, but the former are to be
preferred. They can be easily made, in any number required, by
628 APPENDICES.
cementing a little piece of plate-glass at either end of a glass
slip (Fig. 259).
Glass Rods.—One dozen or more glass rods, twelve to eighteen
inches in length. They are employed for smoothly spreading out
the liquefied nutrient gelatine or agar-agar on the glass plates, etc.
Thermometers.—Two or three centigrade thermometers.
(I) Apparatus FoR PREPARATION OF PoTATO-CULTIVATIONS,
Israel’s Case.— Sterilising instruments in the flame of a Bunsen
burner is most destructive. It is better, therefore, to have a sheet-
iron case (Fig. 260) to
contain potato-knives,
scalpels and other in-
struments, and to ster-
ilise them by placing
the case in the hot-air
steriliser for an hour
at 150° C. The box
can be opened at the
Fic. 260.—Isragu's Case. side, and each instru-
ment withdrawn with
a pair of sterilised forceps when required for ase
Glass Dishes.—Several shallow glass dishes are required for
preparing damp chambers for potato-cultivations (Fig. 261). The
upper, being the larger, fits
over the lower, and having
no handle, admits of these
damp chambers being placed,
if necessary, in the incubator
in tiers. The large size may
also be used in the same
way for plate-cultivations, Fie. 261.—Damp CuamBer ror Porato-
Potato Knives.— A Sy tae
common broad smooth-bladed knife set in a wooden handle is sold
for this purpose.
Scalpels.—Half a dozen scalpels, preferably with metal handles
may be kept especially for inoculating sterilised potatoes.
Brush.—A common stout nail-brush, or’ small scrubbing-brush,
is essential for cleansing potatoes.
i
APPARATUS, MATERIAL, AND REAGENTS. 629
(J) APPARATUS FOR PREPARATION oF SoLIDIFIED STERILE
Bioop-sERUM.
Glass Jar.—A tall cylindrical glass jar, on foot, with a broad
ground stopper, for receiving blood.
Pipette.—An ordinary or graduated pipette, for transferring the
serum from the jars to sterile test-tubes or glass capsules.
Koch’s Serum Ster-
iliser.—A cylindrical case,
with double walls forming an
interspace to contain water,
closed with a lid, also double-
walled and provided with a
tubular prolongation of the
enclosed water-chamber (Fig.
262). The water in the
cylinder is heated from below,
and that in the lid by means
of the prolongation.
In the centre of the
cylinder is a column which
communicates with the water-
chamber of the cylinder, and
from it pass four partitions,
which serve to support the
test-tubes. Fic. 262.—Kocw’s Serum STERILISER.
In the lid are three
openings, one of which communicates with the water-chamber in the
lid by which the latter is filled, and into which a thermometer is
then fixed. In the centre an
opening admits a thermo-
meter, which passes into the
central pipe of the cylinder ;
through a third opening a
thermometer passes to the
cavity of the cylinder. The
cylinder and cover are jacketed
with felt, and the apparatus
is supported on iron legs.
Koch’s Serum Inspis-
sator.—A shallow tin case
Fic. 263.—Srerum INSPISSATOR.
with glass cover, both case and cover jacketed with felt (Fig. 263)
630 APPENDICES.
The case is double-walled, and the water contained in the interspace
is heated from below. It is supported on four legs, and the two
front ones move in grooves in the case, so that the latter can be
placed obliquely at the angle required and secured in position by
screw-clamps. It is employed for coagulating sterile liquid serum,
and for solidifying nutrient agar-agar so as to give them a sloping
surface. 7
Hueppe’s Serum Inspissator.—By the new process the serum
is obtained with every possible precaution, and solidified at once in
Hueppe’s apparatus (Fig. 44).
Glass Capsules.—Small capsules or hollowed-out cubes of
crystal glass are employed for cultivation on solid blood-serum, on
nutrient gelatine, and on agar-agar. They may be, procured of
white and blackened glass, and are provided with glass slips as
covers. :
(K) APPARATUS FoR STORING, AND FoR CuLtivarions 1n, Liquip
Mepia.
Lister’s Flasks.—Lister devised a globe-shaped flask with two
necks—a vertical anda lateral one, The lateral one is a bent spout,
tapering towards its constricted extremity. When the vessel is
restored to the erect position after pouring out some of its contents,
a drop of liquid remains behind in the end of the nozzle, and
prevents the regurgitation of air through the spout. A cap of
cotton wool is tied over the orifice, and the residue in the flask kept
for future use. The vertical neck of the flask is plugged with
sterilised cotton wool in the ordinary way (Fig. 60).
Sternberg’s Bulbs.—Sternberg advocates the use of a glass
bulb, provided with a slender neck drawn out to a fine point and
hermetically sealed (Fig. 62).
Aitken’s Test-tube.—This is an ingenious device for counter-
acting the danger of entrance of atmospheric germs on removal from
the ordinary test-tube of the cotton-wool plug. Each test-tube is
provided with a lateral arm tapering to a fine point, which is
hermetically sealed (Fig. 62).
Drop-culture Slides.—-About a dozen or more thick glass
slides with a circular excavation in the centre are required for
drop-cultures (Fig. 48).
Vaseline.—A small pot of vaseline with a camel’s-hair brush
should be reserved especially for use in the preparation of drop-
cultures.
APPARATUS, MATERIAL, AND REAGENTS, 631
Bulbed Tubes.—Glass vessels, such as test-tubes, flasks
and pipettes, which are used in dealing with liquid media, have
already been mentioned under other headings; but bulbed tubes,
Pasteur’s bulbs, and various other forms are also required for special
experiments,
(L) Apparatus For IncuBATION.
There are several forms of incubator, each of which has its
advocates. They are mostly rectangular chests, with glass walls
front and back, or in front y
only. A cylindrical model
is preferred by some. Two
only will be described
here—D’Arsonval’s and
Babes’. The former admits
of very exact regulation of
temperature, and the latter
is a very practical form for
general use.
D’Arsonval’s Incu-
bator.—The “twee
DArsonval” (Fig. 264) is
a very efficient apparatus,
and is provided with a heat-
regulator, which enables
the temperature to he
maintained with a mini-
mum variation. It consists
of a cylindrical copper
vessel, with double walls,
enclosing a wide interspace
for containing a _ large
volume of water. The roof =
of the water-chamber is Fic. 264.—D’Arsonvat’s IncupaTor.
oblique, so that the wall
rises higher on one side than on the other. This admits of the inter-
space being completely filled with water. At the highest point is
an opening fitted with a perforated caoutchouc stopper, through
which a glass tube passes. The mouth of the cylinder itself is
horizontal, and is closed by a lid, which is also double-walled to
contain water. In the lid are four openings: one serves for filling its
632 APPENDICES.
water-chamber, aud the others for thermometers and for regulating
the air supply in the cavity of the cylinder. The cylinder is con-
tinued below by a cone, also double-walled, and there is a perforated
grating at the line of junction of the cylinder and cone. The cone
terminates in a projecting tube provided with an adjustable
ventilator. The apparatus is fixed on three supports united to
one another below. One of them is utilised for adjusting the height
of the heating apparatus. Situated above this leg is the heat-
regulating apparatus (Fig. 265), attached to a circular, lipped
aperture in the outer wall of the
incubator. To the lip is fixed with
six screws the corresponding lip of
a brass box, with a tightly-stretched
diaphragm of indiarubber inter-
vening. Thus the diaphragm
separates the cavity of the box from
the water in the interspace of the
incubator. The cap of the box,
Fic. 265.—Scuiosine’s MEMBRANE wel sonews 4 as bored im the
REGULATOR. centre for the screw-pipe, by which
the gas is supplied. Another pipe
entering the box from below is connected with the gas-burners.
Around the end of the screw-pipe a collar loosely fits, and is pressed
against the diaphragm by means of a spiral wire spring. Close
to the mouth of the screw-pipe a small opening exists, so that the
gas supply to the burners is not entirely cut off even when the
diaphragm completely occludes the mouth of the screw-pipe.
To work the apparatus the tube and plug must be removed, and
the water-chamber filled completely with distilled or rain water at the
temperature required. The caoutchouc plug is replaced and the tube
placed in position. Gas enters through d (Fig. 265), and passes through the
opening at its extremity into the chamber of the box. Thence it passes
through the vertical exit which is connected with the gas-burners. As
the temperature rises the water rises in the tube, and at the same time
exercises a pressure on every part of the walls of the incubator, and
hence on the diaphragm. In consequence of this, the diaphragm bulging
outwards approaches the end of the tube d, and gradually diminishes the
gas supply. Asa result the temperature falls, the water contracts and
sinks in the tube, and the diaphragm receding from d, the gas supply
is again increased. By adjusting the position of the tube d to the
diaphragm, any required temperature within the limits of the working of
the apparatus can be regulated to the tenth of a degree—provided (1)
that the gas supply is rendered independent of fluctuations of pressure
APPARATUS, MATERIAL, AND REAGENTS. 633
by means of a gas-pressure regulator ; (2) that the height of the water
in the tube is controlled daily by the
withdrawal or addition of a few drops of ca
distilled water; and (3) that the apparatus |
is kept in a place with as even a tempera-
ture as possible, and sheltered from currents
of air.
The burners in Fig. 264 are protected
with mica cylinders similar to the burner
represented in Fig. 266. The flames of G2
these burners can be turned down to the
smallest length without danger of extinction,
and the temperature may be regulated very
satisfactorily without using the heat-regulator
just described, if the gas first passes through =
a pressure-regulator (Fig. 269). To provide
against the danger resulting from accidental
extinction of the gas, Koch has devised a
self-acting apparatus (Fig. 267), which, simultaneously with the extinction
of the flame of the burner, shuts off the supply of gas. :
IG. 266.—GAS-BURNER
PROTECTED WITH Mica
CYLINDER.
Fy |
a tll
Il
eran
BF)
Fic. 267.—Kocw’s Sarety BuRNER.
Babés’ Incukator.—The pattern used by Babés is a very
simple one, and may be recommended for economy and efficiency
(Fig. 268).
It consists of a double-walled chest with sides and roof jacketed
with felt. Water fills the interspace between the walls, and on
the roof are two apertures—one for a gas-regulator and the other
634
for a thermometer.
Fic. 268.—Basks’ IncuBATOR.
APPENDICES.
In front the chest is closed in by a sheet
of felt, a glass door, and a sliding
glass panel. The apparatus can be
suspended on the wall or supported
on legs, and is heated from below
by means of protected burners.
The gas should pass first through
a pressure-regulator, and then
through a thermo-regulator to the
burners.
Moitessier’s Gas - pressure
Regulator.—This apparatus is best
explained by reference to the dia-
gram (Fig. 269). In the bottom of
the cylinder (A) are the entrance (h)
and exit (2) gas-tubes. The tap (m)
regulates the size of the flame. The
cover (x 7) roofs in the cylinder (4).
The bell (8) supports, by means of
e and f, the ball valve (d), which
The gas,
entering by &, passes through the
lies in the cover (c c).
valve (d), and is thence conducted by the tube @ to the tube 7. The
bell (B) and the weighted dish (A) are screwed on to the connecting-
rod (g). To diminish as
much as_ possible the
friction of g in 2, g
only touches z~ by three
projecting ridges. Section
of i and g is shown at s.
To put the apparatus in
use it is first levelled, then
h is screwed off, and the
cover (m ) removed. A
mixture of two parts of
pure acid-free glycerine to
one of distilled water is
poured into the cylinder
until it flows out at gq,
which is then closed, and
the cover (n 1) replaced.
Fic. 269.—MoIressigr’s GAS-PRESSURE.
REGULATOR.
The manometers are filled with coloured water, and & and 1
APPARATUS, MATERIAL, AND REAGENTS. 635
connected with the entrance and exit gas tubing respectively. The
pressure of the incoming gas raises the bell (B) ; and with it the valve
(d) is raised towards the opening at cc. The weight (h), which is
replaced on g, by its downward pressure counteracts this upward
pressure of the gas and opens the valve (cc). Thus the flame is
best regulated in the morning, when the pressure is at a minimum;
then supposing an increase of pressure occurs, the weight of /, is
overbalanced, B is raised, and with it d,and the gas supply pro-
portionately diminished by the gradual closing of the valved opening.
Reichert’s Thermo-regulator.—This regulator (Fig. 270)
consists of three parts—a hollow T-piece, a stem and a bulb.
The T-piece fits like a stopper in the upper widened
portion of the stem. One arm of the T is open
and connected with the gas supply; the vertical
portion terminates in a small orifice, and is also
provided with a minute lateral opening. The stem is
provided with a lateral arm, and this arm, the stem,
and the bulb contain mercury. The regulator is
fixed in the roof of the incubator, so that the bulb
projects either into the interior of the incubator or \
into the water-chamber. When the incubator reaches
the required temperature, the mercury is forced up
by means of the screw in the lateral arm, until it Fie. 270.—
closes the orifice at the extremity of the vertical ar
portion of the JT. The gas which passes through ggcprator.
the lateral orifice is. sufficient to maintain the
apparatus at the required temperature. If the temperature of the
incubator falls, the mercury contracts, and gas passing through the
terminal orifice of the JT increases the flame of the burner, and
the temperature is restored.
Page’s Thermo-regulator resembles the above, but instead
of the T-piece there are two pieces of glass-tubing. The outer
tubing envelops the upper part of the stem of the regulator, and
admits of being raised or lowered. The upper end of this tubing
is closed by a cork, which is perforated to admit the narrow glass-
tubing, which represents the vertical arm of the T, passing within
the stem of the regulator. This has a terminal and a lateral
opening, and is the means of entrance for the gas. This regulator
is adjusted by noting when the thermometer indicates the desired
temperature, and then pushing down the outer tube until the
terminal opening of the inner tube, which is carried down with
it, is obstructed by the mercury.
636 : APPENDICES.
Meyer’s Thermo-regulator is represented in Fig. 271. No. I.
shows the construction of the regulator : its inner tube terminates
in an oblique opening, and is also provided with a minute lateral
perture, which prevents the complete shutting off of the gas supply.
=| ————S
Sao
BPR ee eee oe oe
Fic. 271.—Meyer’s THERMO-REGULATOR.
No. IT. illustrates the method of introducing the mercury by suction
through a filling tube, which is substituted for the inner tube of the
regulator. No. III. represents Frinkel’s modification of the same
instrument.
(M) Inocutatine anp Dissectina InsrrumENTS AND APPARATUS
In Common Ussz.
Mouse-cages.—As mice are the animals most frequently
employed for experimental purposes, mouse-cages have been
APPARATUS, MATERIAL, AND REAGENTS. 637
especially introduced, consisting simply of a cylindrical glass jar
with a weighted wire cover.
Dressing-case.—A small surgical dressing-case, with its usual
accessories—forceps, knives, small, straight and curved scissors,
needles, silk, and so forth—will serve for most purposes.
Pravaz’ Syringe.—Koch’s modification of Pravaz’ syringe
admits of sterilisation by exposure to 150° C. for a couple of hours.
Special Instruments and Material.—Instruments required
for special operations, and the materials necessary for strict anti-
septic precautions, need not be detailed here.*
Dissecting-boards.—Slabs of wood in various sizes, or gutta-
percha trays, provided with large-headed pins, are employed for
ordinary purposes.
Dissecting-case.— A dissecting-case, fitted with scalpels,
scissors, hooks, ete., should be reserved entirely for post-mortem
examinations.
Fic. 272.—SipHon Borris, witH FLEXIBLE Tube, Guass NozzLE, AND A
Mour’s PINcHcock.
(N) Genera Laroratory ReEqQuisitss.
Siphon Apparatus.—Two half-gallon or gallon glass bottles,
with siphons connected with long flexible tubes provided with
glass nozzles and pinchcocks (Fig. 272), should be employed for the
* Vide Cheyne, Antiseptic Surgery. 1882.
638 APPENDICES.
following purposes :—One is used to contain distilled water, with
the nozzle hanging down conveniently within reach of the working
table; the other is to contain a solution of carbolic acid (1 in 20),
and may be placed so that the nozzle hangs close to the lavatory
sink or basin. The former replaces the use of the ordinary wash-
bottle, in washing off surplus stain from cover-glasses, etc., and the
latter is conveniently placed for disinfection of vessels and hands
after cleansing with water. They should be placed on the top of a
cupboard or on a high shelf.
Desiccator.—The desiccator (Fig. 273) consists of a porcelain
pan containing concentrated
sulphuric acid and covered
over with a bell-glass receiver.
The sheet of plate-glass upon
which the pan rests is ground
upon its upper surface, and
the rim of the glass bell is
also ground and well greased.
In the centre of the pan is a
column supporting a circular
frame, which is covered with
wire gauze. Slices of potatoes,
upon which micro-organisms
have been cultivated, are
rapidly dried by the action of
sulphuric acid in confined air.
A detailed description of other kinds of apparatus commonly in
use in a research laboratory—-such as the various forms of apparatus
for filtering cultures in liquids, and the reagents necessary for
special chemical investigations—must be sought for elsewhere.
Much information may be obtained about the most recent improve-
ments in bacteriological, chemical and physical apparatus by
reference to manufacturers’ catalogues.*
mei
lL
Fie. 273. —DesiccaTor.
* All bacteriological apparatus may be obtained from Berlin from Dr.
Muencke, 58, Louisen Strasse, or Dr. Hermann Rohrbeck, 24, Karlstrasse.
Dr. George Griibler, Leipzig, is recommended for special staining reagents.
In London, chemicals and bacteriological apparatus can be obtained from
Becker & Co., Hatton Wall, or from Baird & Tatlock, 14, Cross Street, Hatton
Garden, E.C. Mr. Baker, of High Holborn, W.C., is recommended as the
agent for microscopes and objectives by Continental makers, including
Zeiss’ apochromatic objectives.
APPENDIX V.
BIBLIOGRAPHY
CHAPTER I.
HISTORICAL INTRODUCTION.
Andry, De la Génération des Vers dans le Corps de l’Homme, 1701. Charlton
Bastian, Proc. Royal Soc. 1872. Bonnet, Considérations sur les Corps orga-
nisés, 1768. Davaine, Compt. Rend., T. lviii. and lix. Gleichen, Dissertation
sur la Génération, 1778. Hill, Essays in Natural History and Philosophy.
Kircher, Ars magna lucis et umbre, 1646. Koch, Beitrige zur Biologie der
_Pflanzen, 1876. Lister, Pharmaceut. Journal and Transact., 1877. Miiller,
Animalia infusoria, 1786. Pasteur, Compt. Rend., 1859, 1880; Etude sur la
Maladie des Vers-d-soie, 1870. Plenciz, Opera Medico-Physica, 1762. Schréder
and Von Dusch, Ann. der Chem. und Pharm, vol. lxxxix. Schiiltze, Poggen-
dorff’s Ann., vol. xxxix. Schwann, Poggendorff’s Ann., vol. xli. Tyndall, Essays
on floating matter of the air, 1881.
CHAPTER II.
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA.
MoRPHOLOGY.
Baumgarten, Lehrbuch der Pathologischen Mykologie, 1890. Biedert,
Virch. Archiv, Bd. 100, 1885, Billroth, Unters. iiber d. Veg. Form der Cocco-
bacteria septica, 1874. Brefeld, Botanische Untersuch. tiber Schimmelpilze,
Heft 1, 1881. Cohn, Beitriige zur Biologie der Pflanzen, Bd. I., 1872, 1875.
Cornil and Babés, Les Bactéries, 1885. Dallinger and Drysdale, Monthly Micro-
scop. Journ., 1875. De Bary, Verg. Morph. und Liolog. der Pilze, Mycetozoen,
und Bacterien, 1884; Vorlesungen tiber Bacterien, 1885. Dujardin, Histoire
Naturelle des Zoophytes, 1841. Ehrenberg, Die Infusionsthierchen als Volkom.
Organism, 1838. Fisch, Biolog. Centralbl., V., 1885. Fliigge, Handbuch
der Hygiene, 1883; Die Micro-organismen, 2nd Edition, 1886. Gram,
Fortsch der Med., IL, No. 6. Grove, Synopsis of the Bacteria and Yeast
Fungi, 1884. Hallier, Die Pflanzlichen Parasiten, 1866. Hauser, Ueber
Faulniss Bacterien, 1885. Hueppe, Die Formen der Bakterien, 1885. Lankester,
Quart. Journ. Microscop. Science, 1873. Leunis, Synopsis d. Pflanzenkunde :
639
640 APPENDICES.
Hanover, 1877. Lister, Quart. Journ. Microscop. Science, 1873. Lutz,
Fortschr. d. Med., 1881. Marpmann, Die Spaltpilze, 1884. Miller, Deut. Med.
Woch., 1884. Nageli, Die Niederen Pilze, 1877. Neelsen, Biol. Centralbl., IIL,
No. 18, 1883. Sachs, Text-book of Botany, 1882. Wan Tieghem, Compt.
Rend., 1879; Traité de Botanique, 1883. Zopf, Die Spaltpilze, 1885.
GENERAL BIOLOGY.
Arloing, Archiv de Physiol., 1886. Bordoni-Uffreduzzi, Fortschr. d. Med.,
1886. Cheyne, Brit. Med. Journ, 1886. Cohn and Mendelsohn, Beitr. z.
Biol. d. Pflanzen, Bd. III., Heft 1. Cortes, Compt. Rend., T. 99, p. 385, 1884.
Downes, Proc, Roy. Soc., 1886. Duclaux, Compt. Rend., 1885. Engelmann,
Arch. f. d. Ges. Physiologie, Bd. 26, 1881; Botan. Zeitg., 1882. Fodor,
Archiv f. Hygiene, 1886. Hauser, Archiv f. Exper. Patholog. u. Pharmacologie,
1886. Hofmann, Allgem. Med. Centralbl., 8. 605. Liborius, Zeitschrift f.
Hygiene, 1886. Nageli, Die Niederen Pilze: Miinchen, 1877; Unters. iiber
Niedere Pilze: Miinchen, 1882. Nencki, Virchow’s Archiv, 1879 ; Beitrige zur
Biol. der Spaltpilze : Leipzig, 1879; Ber. d. Deutschen Chem. Gesellsch., 8.
2605, 1884, Regnard, Compt. Rend., T. 98, p. 744, 1884. Tumas, St. Petersb.
Med. Wochenschr., 1879. Wyssokowitseh, Zeitschrift f. Hygiene, 1886.
CHROMOGENIC BACTERIA.
Babés, Biolog. Centralbl., Bd. 2, 1882. Chabert and Fromage, D’une Altéra-
tion du Lait de Vache, désignée sous le Nom du Lait Bleu, 1880. Charrin,
Communication faite a la Société Anatomique, 1884. Cohn and Miflet, Cohn’s
Beitriige zur Biol. d. Pflanzen, Bd. III., Heft 1, 1879. Eberth, Centralbl. f. d.
Med. Wissensch., 18638. Ehrenberg, Micr. Prodigiosus, Verhandl. d. Berl.
Acad., 1839, Fordos, Compt. Rend. de l’Acad. de Sc., 1860. Frank, Cohn’s
Beitr. z. Biol. d. Pflanzen, Bd. I., Heft 3, 1875. Gessard, De la Pyocyanine
et de son Microbe, 1882 ; Ann. de l'Institut Pasteur, T. iv. Gielen, Mag. f.
Ges, d. Thierheilkunde, 1852. Girard, Unters. tiber Blauen Hiter; Chirurg.
Centralbl., II., 1875; Revue des Sc., T. 5, 1877. Hermstadt, Ueber die
Blaue und Rothe Milch., 1833. Hugues, Echo Vétérinaire, 1884. Klein,
Quart. Journ. of Micr. Sc., Vol. 15, 1875. Lankester, Quart. Journ. of Micr. Sc.,
Vol. 13, 1873, 1876. Liieke, Arch. f. Klin. Chir., 1862. Mosler, Virchow’s
Archiv, Bd. 43, 1868. Neelsen, Cohn’s Beitr. z. Biologie d. Pflanzen, Bd. III.,
Heft 2, 1880. Schréter, Cohn. Beitr. z. Biol. der Pflanzen, Bd. I., Heft. 2,
1872. Steinhoff, Neue Ann. d. Mecklenb. Landw. Ges., 1838. Wernich, Cohn’s
Beitrige zur Biol. d. Pflanzen, Bd. III., Heft 1, 1879. Van Tieghem, Bull. de
la Soc. Bot. de France, 1880,
ZYMOGENIC BACTERIA AND FERMENTATION.
Béchamp, Compt. Rend., T. 60, p. 445, 1865; T. 93,1881. Boutroux, Compt.
Rend., T. 86, 1878. Brefeld, Landwirthsch. Jabresber., Bd. 3, 1874; Bd. 4,
1875; Bd. 5, 1876. Cienkowski, Die Gallertbildungen d. Zuckerrtibensaftes,
1878. Colin, Bull. de l’Acad. de Méd., 1875. Dubrunfaut, Compt. Rend.,
T..73, 1871. Duclaux, Théses présentées 4 Ja Faculté de Paris, 1865.
Dumas, Compt. Rend., T. 75. Nr. 6, 1872; Ann. de Chim, et de Phys., 1874.
Eriksson, Unters. aus. d. Botan.; Institut in Tiibingen, Heft 1, 1881. Feltz
and Ritter, Journ. de l’Anat. et Phys., 1874. Fermi, Centralb. f. Bact.,
1891. Fitz, Berichte d. Chem. Ges., Bd. 6, 8. 48, 1875; Bd. 10, p. 216, 1878;
BIBLIOGRAPHY. 641
Bd. 11, pp. 42 and 498, and Bd. 12, p. 474, 1879; Bd. 13, p- 1309, 1880;
Bd. 15, p. 857, 1882; Bd. 16, p. 844, 1883; Bd. 17, p. 1188,.1884, Fleck,
Ber. d. Chem. Centralst.. Dresden, 1876. Frankland, Cantor Lectures, 1899.
Gessard, De la Pyocyanine et de son Microbe. Guiard, Thése de Paris, 1883.
Hallier, Gihrungserscheinungen, 1867. Hansen, Untersuch. aus. d. Prax. der
‘Gihrungsindustrie, 1890, 1892, Harz, Grundaziige der alkoholischen Gih-
rungslehre, 1877. Hiller, Centralbl. f. d. Med. Wiss., 8. 53,1874. Hofmann,
Aerztl. Verein 2u Wien, Mai 1873; Allgem. Med. Centralbl., §. 605, 1873.
Hoppe-Seyler, Medic.-Chem. Untersuchungen, Heft 4, 1871. Hueppe, Mitth.
a. d. Ges. Amt, Bd. ii., 1884; Deut. Med. Woch., 1884, Jacksch, Zeitschr. f.
Physiol. Chemie, Bd. 5, 1881. Jorgensen, Microorg. of Ferment Trans., 1893.
Karsten, Chemismus der Pflanzenzelle, 1869. Kern, Bull. de la Soc, Imp. des
Naturalistes de Moskau, No, 3, 1881. Krannhals, Deut. Arch. f. Klin. Med.,
Bd. 35, 1884. Ladureau, Compt. Rend., T. 99, p. 877, 1884. Lépine and Roux,
Compt. Rend., T. 101, 1885. Leube, Virch. Arch., Bd. 100, §.,540, 1885. Lex,
Centralbl. f. d. Med. Wiss., §. 291, 1872. Liebig, Verhandl. der Miinchener
Akad. d. Wiss., 1861; 5 Nov. 1869; Ueber Gihrung, Quelle der Muskel-
kraft und Ernaéhrung; Leipzig u. Heidelberg, 1870. Lister, The Pharmae.
Journ. and Transact., 1877. Mayer, Lehrbuch der Gahrungschemie; 2 Aufl.,
1876. Monoyer, Thése de Strassburg, 1862. Miller, Journ. f. Prakt. Chem.,
1860. Musculus, Ber. d. Chem. Ges., 8. 124, 1874; Compt. Rend., T. 78, 1874.
Nageli, Theorie der Gahrung: Miinchen, 1879. Pasteur, Annal. de Chim. et
de Phys., III. Sér., T. 58, 1860; Compt. Rend., 1860, 1861, 1863, 1864, 1871,
1872; Bull. de la Soc. Chim., 1861; Ann. de Chim. et de Phys., T. 64, 1862;
Etudes sur le Vin, 1866; Bull. de l’Acad. de Méd., No. 27, 1876; Etudes sur
la Biére, 1876. Pasteur and Joubert, Compt. Rend., T. 83,1876. Popoff, Botan.
Jahresber., 1875. Prazmowski, Untersuchungen iiber die HEntwicklungs-
geschichte und Fermentwirkung einiger Bakterien, 1880. Richet, Compt.
‘Rend., T. 88, 1879. Scheibler, Zeitschr. f. Riibenzuckerindustrie, 1874.
Schiitzenberger, Die Gihrungserscheinungen, 1874. Sheridan Lea, Journ.
of Physiology, 1885. Trécul, Compt. Rend., T. 61, 1865; T. 65, 1867; Ann.
des Se., Sér. 7, T. 7, 1867. Tyndall, Compt. Rend., T. 58, 1864; Essays on
the Floating Matter of the Air, 1881. Van Tieghem, Compt. Rend., 1864,
1874, 1879, 1880, 1884.
PHOTOGENIC BACTERIA.
Beyerinck, Archiv Neerland, XXIII. Fischer, Zeitschr. f. Hygiene, 1887 ;
Centralbl. f, Bakteriolog., 1888. Forster, Centralbl. f. Bakteriolog., 1887.
Girard, Compt. Rend., 1890. Girard and Billet, Compt. Rend., 1889. Katz,
Centralbl. f. Bakteriol., 1891. Lehmann, Centralbl. f. Bact., 1889. Ludwig,
Centralbl. f. Bact., 1887.
CHAPTER III.
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA.
Arloing, Cornevin and Thomas, Lyon Méd., 1883. Blyth, Proe. Roy. Soc.,
1885. Buchholz, Ueber das Verhalten von Bakterien zu einigen Antiseptics,
1876; Arch. f. Exp. Pathol., Bd. 7,1877. Chairy, Compt. Rend., 1884. Chamber-
‘land and Roux, Compt. Rend., 1883. Chauveau, Compt. Rend., 1883, 1884.
Cheyne, Antiseptic Surgery, 1882. Colin, Compt. Rend., T. 99, ie De la
642 APPENDICES.
€roix, Arch. f, Exp. Pathol., Bd. 13, 1881. Dujardin-Beaumetz, Bull. de l’Acad,
de Méd.-de Paris, 1884, Eidam, Cobn’s Beitr. zur Biol., Bd. I., Heft 3, 1875,
Fischer, Berl. Klin. Woch., 1882. Fischer and Proskauer, Mitth, a. d. Kaiser.
Ges. Amt, Bd. IJ., 1884, Frank, Ueber Desinfection von Abtrittsgruben, 1885.
Frisch, Sitzungsber. d. Wiener Akad., Bd. 75 u. 80, 1877. Giirtner and Plagge,
Deut. Med. Woch., 1885, Haberkorn, Das Verhalten von Harnbakterien gegen
einige Antiseptica ; Dissert. Dorpat., 1879. Handford, Brit. Med. Journ., 1885,
Heydenreich, Compt. Rend., T. 98, 1884. Hoffmann, Experimentelle Unter-
suchungen iiber die Wirkung der Ameisensiiure Diss. Greifswald, 1884.
Hueppe, Mittheilg. a. d. Kaiserl. Ges. Amt, Bd. 1., 8. 341, 1881 ; Deut. Militar-
Grztl. Zeitschr., 1882. Koch, Cohn’s Beitr. zur Biol. der Pflanzen, Bd. II,
Heft 2, 1876 ; Mitth. a. d. Ges, Amt, Bd. I., 8. 234, 1881. Koch and Gaffky,
Arbeit, a. d. K. Gesundh. Amt, 1885. Koch, Gaffky and Loffler, Mitth. a.
d. Kaiserl. Ges. Amt, Bd. I., 8. 322, 1881. Koch and Wolffhiigel, Mitth. a. d.
Kaiserl. Ges. Amt, Bd. I., §. 301, 1881. Konig, Chirurg. Centralbl., 1885.
Laillier, Ann. d’Hygiéne, 1883. Larrivé, L'Eau Oxygénée: Thése de Paris,
1883. Lassar, Deut. Med. Woch., 1880. Lebedeff, Arch. de Physiol. Norm. et
Pathol., 1882. Maly and Emich, Sitzungsber. d. Kais. Akad. d. Wiss. zu Wien..
Jan, 1883. Marié-Davy, Revue d’Hygiéne, 1884. Merke, Virchow’s Archiv,
Bd. 81, 1880. Meyer, Ueb. d. Milchsiiureferment u. sein Verhalten gegen
Antiseptica, 1880. Mignet, Annuaire de Observatoire de Montsouris, 1884-
Miquel, Semaine Médicale, 1883. Moérscheli, Deut. Med. Woch., 1880. 'Nageli,.
Die Niederen Pilze, 1877, Pasteur, Ann. d’Hyg., 1880; La Vaccination Char-
bonneuse, 1883. Perroncito, Arch. Ital. de Biol., 1883. Pictet and Young,.
Compt. Rend., T. 98, 1884. Plaut, Desinfection der Viehstalle, 1884. Reinl,
Prager Med. Woch., Nr.10 u. 11,1885, Rochefort, Herscher, Revue d’Hygieéne,.
1884, Rossbach, Berl. Klin. Woch., 1884. Schede, Sammlung Klin. Vortrige,
Nr. 25,1885. Schill and Fischer, Mitth. a. d. Kaiserl. Ges. Amt, Bd. II. 1884.
Schnetzler, Archiv de Géneve, 1884. Schréter, Cohn’s Beitr. zur. Biol, der
Pflanzen, Bd. I., Heft 3, 1875. Schultz, Deut. Med. Woch., Nr. 17, 1883;
Nr. 24, 1885. Schwartz, Sitzungsber. d. Dorpater Naturf. Ges., 1879, Soyka,
Ber. d. Bayr. Akad. d. Wissensch., 1879. Steinmeyer, Ueber Desinfectionslehre,
1884. Sternberg, Amer. Journ. Med. Soc., 1883; Report of Com. on Disin-
fectants, 1888. Thol, Ueber d. Einfluss nicht aromat. organ, Séuren. auf
Faulniss u. Gaibrung: Diss. Greifswald, 1885. Toussaint, Bull. de l’Acad.,
1880. Tyndall, Phil. Trans. of the Roy. Soc., 1877. Vallin, Ann. d’Hyg., 1877;
Traité des Désinfectants et de la Désinfection, 1883; Les Nouvelles Etuves
& Désinfection: Revue d’Hygiéne, 1883; Ann, d’Hygiene, 1884. Wernicke,
Virchow’s Archiv, Bd. 78, 1879; Diss. Dorpat., 1879. Grundriss der Desin-
fectionslehre, 1880. Wolff, Centralbl. f. d. Med. Wiss., Nr. 11, 1885. Wolff-
hiigel, Mittheilg. a. d. Kais. Ges. Amt., Bd. I., 8. 188, 1881. Wolffhiigel and
Knorre, Mitth. a. d. Kaiserl. Ges. Amt, Bd. 1., 8. 352, 1881.
CHAPTER IV.
CHEMICAL PRODUCTS OF BACTERIA.
Backlisch, Ber. d. Deutsch. Chem. Gesellsch., Bd. 18, 1880. Bergmann,
Das Putride Gift., 1866; Deut. Zeitschr. f. Chirurgie, Bd. I., 1872. Bergmann
and Angerer, Wiirzburger Jubil. Festschr., 1882. Bergmann and Schmiedeberg,.
Med. Centralbl., 1868. Blumberg, Virch. Arch., Bd. 100, 8. 377, 1885. Boeci,
BIBLIOGRAPHY. 643
Centralbl, f. d. Med. Wiss., 1882. Bouchard, Compt. Rend. de Biol., 1882.
Brieger, Zeitschr. f. physiol. Chemie, Bd. 7, 1883; Ber. d. Deutsch. Chem. Ges.,
Bd. 17, 1884; Berl. Klin. Woch., Nr. 14, 1884 ; Ueber Ptomaine, 1885; Weitere
Untersuchungen iiber Ptomaine, 1885; Ueber Ptomaine: Berl. Klin..Woch.,
1886. Brouardel and Boutmy, Compt. Rend., T. 92, p. 1056, 1881. Clementi
and Thin, Wien. Med. Jahrb., 1873. Eber, Centralb. f. Bact., 1892. Etard and
Olivier, Compt. Rend., 1882. Frisch, Exper. Studien iib. d. Verbreitung d.
Faulnissorganismen, 1874. Gautier, Compt. Rend., T. 94, 1882. Gautier and
Etard, Compt. Rend., T. 94, 1882. Groebner, Beitriige z. Kenntniss der
Ptomaine, 1882. Guareschi and Mosso, Arch. Ital. de Biolog., 1883. Hauser,
Ueber Faulnissbakterien, 1885. Hemmer, Exper. Studien iiber d. Wirkung
Faulender Stoffe, 1866. Hiller, Centralbl. f. Chirurgie, 1876 ; Die Lehre von der
Faulniss, 1879. Husemann, Arch. d. Pharmac., 1880, 1882, 1883. Kaufmann,
Journ, f. Prakt. Chemie, Bd. 17, 1878. Kehrer, Archiv f. Exper. Pathol.
Bd. I, 1874. Kénig, Ber. iib d. Veterinirwesen im Kénigreich Sachsen,
1881. Maas, Fortschr. d. Med., II., 729, 1884. Martin, Rep. Med. Off. Local
Govt. Board, 1890-91. Nencki, Ueber die Zersetzung der Gelatine und des
Hiweisses bei der Faulniss mit Pancreas, 1876; Journ. f. Pract. Chem.,
Bd. 26, 1882. Offinger, Die Ptomaine,1885. Otto, Anleitung zur Ausmittelung
der Gifte, 1884. Panum, Virchow’s Arch., Bd. 60, 1874. Raison, Zur Kenntniss
der Putriden Intoxication, 1866. Ravitsch, Zur Lehre von der Putriden In-
fection, 1872. Rosenbach, Deut. Zeitschrift fiir Chir., XVI., 8. 342, 1882.
Salomonsen, Die Faulniss des Blutes, 1877. Schiffer, Arch. f. Anat. u. Physiol. :
Physiol. Abtheil, 1882. Schweninger, Ueber d. Wirkung Faulender Org. Sub-
stanzen, 1866. Selmi, Chemische Ber., Bd. 6, 7, 12, 1878. Tanret, Compt.
Rend., T. 92,1881. Tappeiner, Med. Centralbl., 1885. Vandevelde, Arch. de
Biol. par van Beneden, 1884. Willgerodt, Ueber Ptomaine, 1884. Ztilzer and
Sonnenschein, Berl. Klin. Woch., 1869,
CHAPTER V.
IMMUNITY.
Arloing, Cornevin and Thomas, Du Charbon Bactérien; Pathogénie et
Inoculations Préventives, 1883. Behring, Deutsche Med. Woch., 1890.
Behring and Kitasato, Deutsche Med. Woch., 1890. Blazekovic, Oesterr
Monatschr. f. Thierheilk., 1884. Bouchard, Compt. Rend., 1889. Bouley,
LInoculation Préventive de la Fiévre Jaune: Compt. Rend., T. 100, 1885.
Brieger and Frankel, Berl. Klin. Woch., 1890. Biichner, Eine neue Theorie
iiber Erzielung v. Immunitat gegen Infectionskrankheiten, 1883. Chamberland,
Le Charbon et la Vaccination Charbonneuse d’aprés les Travaux Récents de
M. Pasteur, 1883. Chamberland and Roux, Compt. Rend., T. 96, Nr. 15, 1883.
Chauveau, Compt. Rend., T. 89, 1879; T. 96, Nr. 9; Nr. 10; Nr. 11, 1883; Gaz.
Hebdom. de Méd. et de Chir., 22, 1884. Feltz, Compt. Rend., T. 99, p 246,
1884. Frank, Jahresber. d. K, Thierarzneischule in Miinchen, 1883. Gamaleia,
La Semaine Med., 1890. Grawitz, Die Theorie der Schutzimpfung: Virchow’s
Arch., Bd. 48, 1881. Hankin, Brit. Med. Journ., 1889 and 1890; Proc. Roy.
Soc., 1890; Centralb. f. Bacteriolog., Bd, IX., Lancet, 1891. Hess, Schweiz.
Arch. £, Thierheilk., Bd., 27,1885. Kitasato, Zcitschr. f. Hygiene, Bd. X. Koch,
Ueber die Milzbrandimpfung, 1882. Koch, Gaffky and Loffler, Mitth. a. d. Ges.
Amt, Bd, II., 1884, Léffler, Mitth. a. d. Ges, Amt, Bd. L,1881. Martin, Reports
644 ‘APPENDICES.
Med. Dept. Loc. Govt. Board, 1890-91; Brit. Med. Journal, 1891. Massé, Des
Inoculations Préventives dans les Maladies’ Virulentes, 1883. Metschnikoff,
Virchow’s Archiv XCVI. and XCVIL, Ann, de l'Institut Pasteur, 1887, 1889,
1890, 1891, 1895. Nuttall, Zeitschr. f. Hygiene, 1888. Oemler, Arch. f. Wiss.
u. Pract. Thierheilk., 1876, 1881. Ogata, Centralb. f. Bacteriolog., 1891. Ollive,
Compt. Rend., T. 89, 1879. Pasteur, Bull. de l’Acad. de Méd. and Gaz.
Méd, de Paris, Nr. 18, 1880; Compt. Rend., 1883; La Vaccination Charbon-
neuse, 1883; Revue Scientifique, 1883; Bull. de lAcad. de Méd., 1883.
Perroncito, Atti R. Acc. d. Lincei., 1883. Pitz, Vortrige f. Thierirzte,
Ser. 7, Heft 1, 1884. Roux, Ann. de 1l’Institut Pasteur, 1888. Részahegyi,
Pester Med.-Chir. Presse, 1882. Salmon and Smith, Centralb. f. Bacteriolog.,
1887, Semmer, Virchow’s Arch., Bd. 83, 1881. Semmer and Krajewski,
Centralbl. f. d. Med. Wiss., 1880. Strebel, Schweiz. Arch. f. Thierheilk., 1885.
Tizzoni and Cattani, Centralb. f. Bacteriolog., 1891. Toussaint, Bull. de
VAcad. de Méd.; and Compt. Rend., 1880, 1881; Gazette Médicale de Paris,
Nr. 32, 1881. Wooldridge, Proc. Roy. Soc., 1887; Archiv f. Anat. and Phys.,
1888.
CHAPTER VI.
ANTITOXINS AND SERUM-THERAPY.
Béclére, Chambon and Ménard, Ann. de l'Institut Pasteur, 1896. Behring
and Kitasato, Deutsche Med. Woch., 1890; Trans. Internat. Cong. f. Hygiene,
1891, Behring and Wernicke, Zeitschrift f. Hygiene, 1892. Buchner, Munich
Med. Woch., 1891. Calmette, Ann. de l'Institut Pasteur, 1895. Emmerich and
Mastbaum, Archiv f. Hygiene, 1891. Fedoroff, Zeitschr. f. Hygiene, 1893.
Gromakowsky, Ann. de l'Institut Pasteur, 1895. Heubner, Trans. Internat.
Med. Congress, 1894. Hewlett, Practitioner, 1895. Kossel, Zeitschrift f.
Hygiene, 1894. Marchouz, Ann. de l'Institut Pasteur, 1895. Marmorek, Ann.
de l'Institut Pasteur, 1895, 1896. Ogata, Centralb. f. Bakteriolog., 1891.
Report, Med. Sup. Metropolitan Asylums Board, 1896. Roux, Trans. Internat.
Congress of Hygiene, 1894; Ann. de l'Institut Pasteur, 1894. Roux, Martin,
Chaillou, Ann. de I’Instit. Pasteur, 1894. Tizzoni and Cattani, Centralb. f,
Bakteriolog., 1891. Welch, Trans, Assoc. Amer. Physicians, 1895. Wladmoroff,
Zeitschr, £. Hygiene, 1893.
CHAPTERS VII. VIIL, IX. X.
VII.—THE BACTERIOLOGICAL MICROSCOPE. VIII.—MICROSCOPICAL
EXAMINATION OF BACTERIA, IX.—PREPARATION OF NUTRIENT
MEDIA AND METHODS OF CULTIVATION. X.—-EXPERIMENTS
UPON THE LIVING ANIMAL.
Almquist, Hygeia, XLV.; Stockholm, 1883. Banti, Manuale di Tecnica
Batteriologica, 1885. Baumgarten, Zeitschr. f, Wissensch. Mikr., 1884. Behrens,
Hilfsbuch zur Ausfiihrung Mikroskop. Untersuch,, 1883. Bizzozero and Firket,
Manuel de Microscopie Clinique, 1885. Blanchard, Rev. Inter. Sci., ILL, 1879.
Bordoni Uffreduzzi, Microparasitici, 1885. Brefeld, Bot. Untersuch. iiber
Schimmelpilze, Bd. IV., 1881; Botan. Unters. iiber Hefenpilze, Bd. V., 1883;
Verhandl. d. Physik. Med. Ges. in Wiirzburg, 1875. Biichner, In Nigeli’s
BIBLIOGRAPHY. 645
Untersuch. tiber Niedere Pilze: Munich, 1882; Aerztl. Intelligenzbl., No. 33,
1884. Carpenter, The Microscope and its Revelations (6th Edition), 1881.
Cohn, Beitriige zur Biologie der Pflanzen, Bd. I., Heft 3, 1875, 1876. Cornil
and Babés, Les Bactéries, 1886. Crookshank, Manuel Pratique de Bactério-
logie, traduit par Bergeaud; avec 4 Photomicrographies, 1886. Dolley, Tech-
nology of Bacteria Investigation, 1885. Duclaux, Ferments et Maladies, 1881.
Ehrlich, Deut. Med. Woch., No. 19, 1882 ; Zeitschr. f. Klin. Med.; Bd. I. 1880.
Bd. Il., Heft 3, 1881. Eisenberg, Bakteriologische Diagnostik, 1886. Esmarch,
Zeitschrift f, Hygiene, 1886. Fehleisen, Ueber Neue Method. der Untersuch.
u. Cultur Pathogen. Bakterien ; Physik. Med. Ges. zu Wiirzburg, 1882, Fligge,
Handbuch der Hygiene und der Gewerbe Krankheiten, 1883. Friedlander,
Microscopische Technik (Ist Edition), 1884, Gibbes, Practical Histology and
Pathology, 1885. Gram, Fortsch. d. Med., IL, No. 6, 1884. Hauser, Ueber
Faulnissbacterien; mit 15 Tafeln in Lichtdruck, 1885. Hazlewood, American
Monthly Microscop. Journ., 1883. Hiiber and Breker, Die Path. Histolog. und
Bacteriologischen Untersuch. Methoden, 1886. Hueppe, Bakteriologische
Apparate : Deut. Med. Woch., 1886; Die Methoden der Bakterien Forschung,
1886; translated by Biggs, 1886. Johne, Ueber die Kochschen Reinculturen,
1885. Klebs, Archiv f. Exp. Pathoi., Bd. 1, 1873. Klein, Micro-organisms and
Disease, 1886. Koch, Biol. Klin. Woch., No. 15, 1882; Mitth. a. d. Kais. Ges.
Amt, Bd. IL, 1881.; Bd. II, 1884; Untersuchungen iiber Wundinfections
Krankheiten, 1878; Beitriige z. Biol. d. Pflanzen, Bd. II., Heft 3, 1877. Lee,
The Microtomist’s Vade Mecum,1885. Magnin and Sternberg, Bacteria, 1884.
Malley, Photomicrography, 1885. Orth, Path. Anat. Diagnostik, 1884. Pasteur,
Etudes sur la Biére, 1867. Perty, Zur Kenntniss Kleinster Lebensform,
1852. Plaut, Farbungs Methode z. Nachweis. der Micro-organismen, 1885 ;
Salomonsen, Bot. Zeit., No. 39, 1879; No. 28, 1880. Schafer, Course of Practical
Histology, 1877. Woodhead and Hare, Pathological Mycology, 1885.
CHAPTER XI.
EXAMINATION OF AIR, SOIL AND WATER.
Angus Smith, Rep. to the Loc, Gov. Board, 1884; Sanitary Record, 1883.
Becker, Reichsmedicinalkalender, 1885. Beumer, Deut. Med. Woch., 1886.
Bischof, Journ. Soc. Chem. Industry, 1886, Biichner, Vortrige im Aeratl.
Verein zu Miinchen, 1881. Chamberland, Compt. Rend., T. 99, p. 247, 1886.
Cramer, Die Wasserversorgung von Ziirich, 1885. Crookshank, Notes from a
Bact. Labor., Lancet, 1885. Cunningham, Micr. Exam. of the Air: Calcutta,
1874, Fodor, Hygienische Unters. tiber Luft, Boden u. Wasser., 1882.
C. Frankel, Zeitschr. f. Hygiene, 1886. Frankland, P. and G., Proc. Roy. Soc.;
1885, 1886; Microorganisms in Water, 1894, Gunning, Arch. f. Hyg., 3, 1883.
Hereus, Zeitschr. f. Hygiene, 1886. Hesse, Deut. Med. Woch., Nr. 51, 1884 ;
2, 1884; Mitth. a. d. Ges. Amt, Bd. IL., 1884; Ueber Wasserfiltration : Deut.
Med. Woch., 1885; Zeitschr. f. Hygiene, 1886. Klebs and Tommasi-Crudeli,
Archiv f. Exper. Path., Bd. 11, 1879. Koch, Mitth. a. d. Ges. Amt, Bad. L.,
1881. Laurent, Journal de Pharmacie et de Chimie, 1885. Lemaire, Compt.
Rend., T. 57, 1863. Letzerich, Exp. Unters. iib. die Aetiologie des Typhus
mit bes. Beriicksichtigung der Trink. u. Gebrauchswasser, 1883, Maddox,
Month. Microscop. Journal, 1870. Meade Bolton, Zeitschr. f. Hygiene, 1886,
Miflet, Cohn’s Beitr. z. Biol. d. Pflanzen, Bd, III., 1879. Miquel, Annuaire
*
646 APPENDICES.
de l’Observat. de Montsouris, 1877, 1882; Compt. Rend., T. 86, 1878; Bull.
de la Soc. Chim., 1878; Ann. d’Hygiene, 1879; Les Organismes Vivants
de lAtmosphére, 1883. Miquel and Freudenreich, La Semaine Médicale, 1884,
Moreau and Plantymausion, La Semaine Médicale, 1884. Nageli, Unters. iiber
Niedere Pilze., 1882. Olivier, Les Germes de l’Air, Thése, Rev. Scientif.,
1883. Pasteur, Ann. de Chim. et de Phys., T. 64,1862; Compt. Rend., T. 50,
1860; T. 52, 1861; T. 56, 1863; T. 85, 1877. Pfeiffer, Zeitschr. f. Hygiene,
1886. Pouchet, Compt. Rend., T. 47, 1858. Schrakamp, Archiv f. Hygiene,
Bd. IL, 1884. Sehlen, Fortschr. d. Med., Bd. II., 8. 585, 1885. Smart, Germs,
Dust and Disease, 1883, Soyka, Sitz.-Ber. der. K. Bayr. Akad. d. Wiss, : Math,
Physik. Classe, 1881; Vortriige im Aerztl., Verein in Miinchen, 1881; D.
Vierteljsch. f. Oeff. Ges., Bd. 14, 1882; Prager Med. Woch., 1885; Fortschr,
d. Med., 1885. Tissandier, Compt. Rend., T. 78, 1874. Torelli, La Malaria
in Italia, 1883, Tyndall, Brit. Med. Journ., 1877; Essays on the Floating
Matter of the Air, 1881; Med. Tim. and Gaz., 1870. Wernich, Cohn’s Beitrige
z. Biol. d. Pflanzen, Bd. III., 8. 105, 1879. Wolffhtigel and Riedel, Arbeit.
a. d, K. Ges. Amt, 1886. Wollny, Viert. f£. Oeff. Ges., S. 705, 1883. Zander,
Centralbl. f. Allg. Ges., 1883.
CHAPTER XII.
PHOTOGRAPHY OF BACTERIA.
Crookshank, Photography of Bacteria, 1887. Frankel and Pfeiffer, Mikro-
photog. Atlas, 1889. Giinther, Photogram. Path. Mikroorg., 1887. Itzerott
and Niemann, Atlas der Microphotograph, 1895. Koch, Cohn’s Beitraige zur
Biol. der Pflanz., 1877 ; Mitth. a. d. K. Gesundheitsamte, Bd. 1, 1881. Neuhaus,
Centralbl. f. Bakteriolog., Bd. IV. Sternberg, Photomicrographs and How to
Make them, 1884. Woodward, Rep. to Surg. Gen. U. 8. Army, 1870.
CHAPTER XIII.
SUPPURATION. PYMIA. SEPTICHMIA. ERYSIPELAS. GONORRHGA.
OPHTHALMIA.
Ainstie, Lancet, 1870. Arloing, Recherches sur les Septicémies, 1884.
Babés, Compt. Rend., 1883. Balfour, Edinb. Med. Journ., 1877. Barthold,
Pyzmisch-Metast, Dissert. Berlin, 1875, Bastian, Brit. Med. Journ., 1878.
Béchamp, Compt. Rend., 1881;. Trans. Internat. Med. Cong. London, 1881.
Beck, Rep. Loc. Govt. Board, 1880. Birch-Hirschfeld, Untersuchungen iiber
Pyaimie, 1873. Braidwood and Vacher, Brit. Med. Journ., 1882. Burdon-
Sanderson, Trans. Path. Soc., 1872; Brit. Med. Journ., 1875. Crookshank,
Trans. Internat. Congr. of Hygiene, 1892. Dowdeswell, Quart. Journ. Micr.
Sc. London, Vol. 18, 1878; Proc. Roy. Soc. London, Vol. 34, 1883. Dreschfeld,
Brit. Med. Journ., 1883. Drysdale, Pyrexia, 1880, Garré, Fortschritte d.
Med., 165, 1885. Heiberg, Die Puerperalen u. Pyimischen Processe, 1873.
Hoffa, Fortsch. d. Med., 1885. Horsley, Rep. Med. Officer Loc. Govt. Board,
1881. Klemperer, Zeitschr. f, Klin. Med., 1885. Koch, Wundinfectionskrank-
heiten: Leipzig, 1878; Mittheil. d. Kais. Ges. Amts, Bd. I., 1881. Lister,
Lancet, 1867; Med. Times and Gazette, 1877; Quart. Journ. Microscop. Science,
»
BIBLIOGRAPHY. 647
1887. Oertel, Zur Aetiologie der Infectionskrankheiten, 1881, Ogston, Brit,
Med. Journ., Vol. I., 1881; Journ. of Anat. and Phys., Vol. 17, 1882. Passet,
Ueber Mikroorganismen der Eitrigen Zellgewebsentziindung des Menschen ;
Fortschritte d. Med., Nr. 2, 1885, Bd. 3, 1885. Perret, De la Septicémie :
Paris, 1880. Rindfleisch, Lehrb. der Pathol. Gewebelehre: 1 Aufl. S. 204,
1866. Rosenbach, Mikrooganismen bei den Wundinfectionskrankheiten des
Menschen: Wiesbaden, 1884. Sternberg, Amer. Journ. Med. Sc.; Johns
Hopkins Univ. Stud. Biol. Lab., 1882. Steven, Glasgow Med. Journal, 1884.
Sutton, Trans. Path. Soc., 1883. Tiegel, Ueber d. Fiebererregenden Higen-
schaften des Microsporon Septicum: Bern. Diss., 1871; Virchow’s Archiv, Bd.
60, 1874. Waldeyer, Virchow’s Arch., Bd. 40, 1867 ; Vortrag. i. d. Med. Ges.
zu Breslau, 1871. Watson-Cheyne, Trans. Path. Soc., xxxv., 1884.
OSTEOMYELITIS,
Becker, Deut. Med. Woch., and Berl. Klin. Woch., 1883. Collmann, Bak-
terien im Organismus eines an einer Verletzung am Oberschenkel verstorbenen
Miadchens: Gottingen, 1873. Colzi, Lo Sperimentale, 1890. Courmont and
Jaboulay, Compt. Rend. Soc. de Biolog., 1890. Eberth, Virchow’s Arch.,
Bd. 65, 1875. Fehleisen, Phys. Med. Ges. Wiirzburg, 1882. Friedmann, Berl.
Klin. Woch., 1876. Garré, Fortschr. d. Med., 1885. Giordano, Prog. I. Mic.
Pyog. infett. u. Eziolog. d. Osteom, Impett. Acuta, 1888, Krause, Fortschr. d,
Med., Bd. 2, 1884. Lannelongue and Achard, La Semaine Med., 1890; and
Compt, Rend. Soc. de Biolog., 1890. Peyroud, Compt. Rend., 1884. Rodet,
Compt. Rendus, T. 99, 1884. Rosenbach, Centralbl. f, Chirurgie, 1884.
Schiiller, Centralbl. f. Chirurgie, Nr. 12, 1876.
ENDOCARDITIS.
Birch-Hirschfeld and Gerber, Archiv d., Heilkunde, 1876. Bramwell,
Diseases of the Heart, 1884. Bristowe, Brit. Med. Journ., 1884, Coupland,
Brit. Med. Journ., 1885. Gibbes, Brit. Med. Journ., 1884. Goodhart, Trans.
Path. Soc., vol. xxxi., 1880, Hamburg, Berlin: Inaug.-Diss., 1880. Klebs,
Archiv f. Exper. Pathol., Bd. 9, 1878. Koch, Mittheil. a. d, Kais, Ges. Amt,
Bd. I., 1881. Koester, Virchow’s Arch., Bd. 72, 1875. Kundrat, Sitz.-Ber.
d. Kais. Acad. d. Wissensch. zu Wien, 1883. Leyden, Zeitschr. f. Klin. Med.,
1881. Nocard, Recueil de Méd. Vét., 1885. Oberbeck, Casuistische Beitrige
zur Lehre von der Endocarditis Ulcerosa : Inaug.-Diss., 1881. Orth, J., Ver-
sammlung Deutscher Naturf. zu Strasburg, 1885. Osler, Brit. Med. Journ., 1885 ;
Trans. Int. Med. Congress, 1881. Ribbert, Fortsch. d. Med., 1886. Rosenbach,
Archiv fiir Exper. Pathol., Bd. 9, 1878. Wedel, Berl. Klin. Wochenschr., 1877.
Weichselbaum, Wien. Med. Woch., 1885. Weigert, Virchow’s Arch., Bd. 84,
1881. Wilks, Brit. Med. Journ., 1882. Wyssokowitech, Centralbl f. d. Med.
Wissensch., Nr. 33, 1885.
ERYSIPELAS.
Baader, Schweiz. Naturf. Gesellsch., 1875. Denuce, Etude sur la Pathogénie
et VAnatomie Pathologique de VErysipéle, 1885. Dupeyrat, Recherches
Cliniques et Expérimentales sur la Pathogénie de YErysipele, 1881. Fehleisen,
Wiirzburger Phys. Med. Ges., 1881; Deut. Zeitschr. f. Chir., Bd. 16, 1882; Die
Aetiologie des Erysipels. : Berlin, 1883. Hiiter, Med. Centralbl., Ny. 34, 1868.
648 -APPENDICES.
Janicke and Neisser, Centralbl. f. Chir., Nr. 25, 1884. Klebs, Archiv f. Exper:
Pathol. u. Pharmacol., Bd. 4, 1875, Lukomsky, Virchow’s Archiv, Bd. 60, 1874.
Nepven, Des Bactéries dans l’Erysipéle, 1885. Orth, Archiv f. Exper. Pathol.
u. Pharmacol., Bd. I., 1873. Raynaud, Union Méd., 1873, Recklinghausen and
Lankowski, Virchow’s Arch., Bd. 60, 1874. Rheiner, Virchow’s Arch., Bd. 100,
Heft 2, 1884. Tillmanns, Verhandl. d. Deutsch. Ges. f. Chirurgie, 1878;
Archiv f, Klin. Chirurgie, Bd. 23, 1879. Trogsier, Bull. Soc. Anat. de Paris,
1875. Wolff, Virchow’s Arch., Bd. 81, 1880.
PUERPERAL FEVER.
Aufrecht, Naturforsch. Versamml., 1881. Doléris, La Fiévre Puerpérale et:
les Organismes Infect., 1886. Heiberg, Die Puerperalen und Pydmischen
Processe, 1873. Karewski, Zeitschr. f. Geburtsh. u. Gynadkologie, Bd. 7, 1881.
Laffter, Bresl. Aerztl. Ztg., 1879. Mayrhofer, Monatsschr. f. Geburtsk. u.
Frauenkrankheiten, Bd. 25, 1865. Orth, Virchow’s Arch., Bd. 58, 1873.
Pasteur, Bull. de Acad. de Méd., T. 9, 1880. Recklinghausen and Lankowski,.
Virchow’s Arch., Bd. 60, 1873. Waldeyer, Arch. f. Gynakologie, iii., 1872.
GONORRHGA.
Arning, Viertelj. f. Dermatol. u. Syph., 8. 371, 1883. Aufrecht, Patho-
Jogische Mittheilungen,-1881; Centralbl. f. d. Med. Wiss., Nr. 16, 1883,
Bockhart, Sitzungsbericht d. Phys. Med. Ges. zu Wiirzburg, 1882; Viertelj. f.
Dermatol. und Syph., 1888. Bokai, Allgem. Med. Centralzeitung, Nr. 74,
1880. Bokai and Finkelstein, Prager Med. Chir. Presse, 1880. Biicker,.
Ueber Polyarthritis Gonorrhoica. Diss., 1880. Bumm, Der Mikroorganismus
der Gonorrhoischen Schleimhauterkrankungen, 1885. Campona, Italia Medica,
1883. Chameron, Progrés Medical, 43, 1884. Eschbaum, Deut. Med. Woch.,
8. 187, 1883. Frankel, Deut. Med. Woch., Nr. 2, 1883; S. 22,1885. Haab,
Der Mikrokokkus der Blennorrhcea Neonator, 1881. Hirschberg and Krause,
Centralbl. f. Pract. Augenheilk., 1881. Kammerer, Centralbl. f. Chirurgie,
Nr. 4, 1884. Krause, Die Mikrokokken der Blenorrhcea Neonator, 1882.
Kroner, Naturforschervers. in Magdeburg, Arch. f.'Gyn., xxv., 8. 109,.1884..
Leistikow, Charité-Annalen, 7 Jabrg., 8. 750, 1882. Lundstrém, Studier dfver
Gonokokkus : Diss. Helsingfors, 1885. Martin, Rech. sur les Inflamm. Métast.
& la Suite de la Gonorrhée, 1882, Neisser, Centralbl. f. d. Med.- Wiss.,
Nr. 28, 1879; Deutsche Med. Woch.,:1882. Newberry, Maryland Med. Journ.,
1883. Petrone, Rivista Clin., No. 2. Reter, Centralbl. f. d. Med. Wiss., 1879.:
Sanger, Naturforschervers, in Magdeburg ; Ibid., 8. 126, 1884. Schrétter and
Winkler, Centralbl. f. Bact., Bd. ix. Smirnoff, Vrach., 1886. Steinschneider,.
Verhandl. der Deutsch. Dermat. Gesellsch., 1889; Beri. Klin. Woch., 1890.
Sternberg, Med. News, Vol. 45, Nr. 16, 1884. Weiss, Le Microbe du Pus
Blennorrhagique, 1889. Welander, Gaz. Med. de Paris, 1884. :
OPHTHALMIC DISEASES.
Balogh, Med. Centralbl., xiv., 1879. Bock, Virchow’s Arch., Bd., 91, 1883.
Cornil and Berlioz, Compt. Rend. de l’Acad. d. Sc., 1883. Deutschmann,’
v. Graefe’s Archiv, Bd. xxxi., 1885. Gifford, Archiv f. Augenheilkunde, 1886:
Goldzieher, Centralblatt f. Prakt. Augenheilk., 1884. Kahler, Prager Zeitschr,.
£. Prakt, Heilk., Bd. ‘I, 1882, Klein, Centralbl. f. d: Med. Wiss., Nr. 8, 1884:
BIBLIOGRAPHY. 649
Kroner, Verh. d. Naturforscher-Vers. Magdeburg, 1884. Kuschbert, Deutsch.
Med. Woch., Nr, 21, 1884. Kuschbert and Neisser, Bresl. Aerztl. Zeitschr.,
Nr. 4, 1883. Michel, Graefe’s Archiv f, Augenheilkunde, 1882. Neisser,
Fortschr. d. Med., Bd. 2, 8. 73, 1884. Reuss, Wien. Med. Presse, 1884.
Roth, Virchow’s Archiv, Bd. 55, 1872. Salomonsen, Fortschr. d. Med., Bd. 2,
8. 78, 1884. Sattler, Ber. iib. d. Ophthalmologen Congress zu Heidelberg,
1882; Zehender’s Klin. Monatsblatt, and Wien. Med. Woch., Nr. 17, 1883.
Sattler and de Wecker, L’Ophthalmie Jéquiritique, 1883. Schleich, Verh. des
Ophthalmologen Congr. zu Heidelberg, 1883. Vennemann and Bruylants, Le
Jéquirity et son Principe Pathogéne, 1884. Vossius, Berl. Klin. Wochenschr.,
Nr. 17, 1884. Widmark, Hygeia: Stockholm, 1885. Zehender, Bowman
Lecture: Lancet, 1886.
CHAPTER XIV.
ANTHRAX.
Archangelski, Centralbl. f. d. Med. Wiss., 1882, 1883. Bert, Compt. Rend.
Soc. de Biol., T. 4, 1877; T. 5, 1878; T. 6, 1879. Bleuler, Correspondenzbl. a.
Schweiz Aerzte, 1884. Bollinger, Arbeit. a. d. Patholog. Inst. zu Mitinchen,
1886; Centralbl. f d. Med. Wiss., Bd. 10, 1872; Sitzungsber. d. Ges. f.
Morphol. Physiol. zu Miinchen, 1885. Bouley, Bull. Acad. de Méd.. Paris,
JT. 9, 1880; T. 10,1881; Compt. Rend., T. 92 and 93, 1881. Brauell, Virchow’s
Arch., Bd. 11, 1857; Bd. 14, 1858. Biichner, Ueber die Exper. Erzeugung
des Milzbrandcontagiums aus den Heupilzen, 1880; Sitgungsber. d. K. Bayer.
Akad. d. Wissensch., 1880; Vortrige im Aerztl. Verein zu Miinchen, 1881;
Virchow’s Arch., Bd. 91, 1883. Chauveau, Compt. Rend., T. 90 and 91, 1880;
T. 92, 1881; T. 94, 1882; T. 96, 1883. Chelechowsky, Der Thierarzt, 1884. Colin,
Bull. Acad, de Méd.: Paris, T. 2, 1873; T. 7, 1878; T. 8, 1879; T. 9, 1880;
T. 10, 1881. Crookshank, Rep. Agric. Dept., 1888. Davaine, Compt. Rend.:
Paris, T. 77, 1873; T.57 and 59, 1863 ; T. 60, 1865 ; T. 61, 1866, 1877 ; Rec. de
Méd. Vét., T. 4,1877. Dowdeswell, Rep. Med. Off. Local Gov. Board, 1883. Esser
and Schiitz, Mitth. a K.*Preuss Amt]. Vet. Sanitatsbericht, 1882. Ewart,
Quart. Journ. of Microsc. Sc., 1878. Feltz, Compt. Rend., T. 95, 1882. Fodor,
Deut. Med. Woch., 1886. Fokker, Centralbl. f. d. Med. Wissensch., Bd. 18,
1880; Centralbl. f, d. Med. Wiss., 1881; Virchow’s Archiv, Bd. 88, 1882.
Frank, Zeitschr. f. Hygiene, 1886. Friedrich, Zur Aetiologie des Milzbrands.,
1885. Greenfield, Quart. Journ. Micr. Sc. London, 1879; Proc. Roy. Soc.
London, 1880. Hoffa, Die Natur des Milzbrandgiftes: Wiesbaden, 1886.
Huber, Deut. Med. Woch., Bad. 7, 1881. Johne, Ber. ii. d. Veter. Wesen. i. K.
Sachsen, 1886. Kitt, Sitzungsb. d. Ges. f. Morphol. u. Physiol. zu Miinchen,
1885. Klein, Rep. of the Medical Officer of the Local Govt. Board, 1881; Quart.
Journ. Micr. Sc., 1883. Koch, Beitrage zur Biologie der Pflanzen, Bd. IL.,
Heft 2, 1876; Wundinfectionskrankheiten, 1878; Mitth. aus d.-Ges. Amt,
Ba. 1., 1881 ; Milzbrand und Rauschbrand : Stuttgart, 1886. Martin, Proc. Roy.
Soc., 1890; Rep. Med. Off. Local Govt. Board, 1890-91; Brit. Med. Journal,
1891. Oemler, Archiv f. Wiss. u. Pract. Thierheilk., Bd. 4, 1878, 1879, 1880.
Osol, Experiment. Untersuch. ii. das Anthraxgift: Inaug. Diss. Dorpat, 1885.
Pasteur, Bull. Acad. de Méd., 1877, 1879, 1880 ; Compt. Rend., Paris, T. 84, 1877 ;
T. 90 and 91, 1880; T. 92, 1881; T. 95, 1882. Pollender, Vierteljabrschr.
£. Ger. Med., Bd. 8, 1855. Prazmowski, Acad. d. Wissensch. in Krakau, 1884 ;
650 APPENDICES.
Biol. Centralbl., Bd. 4, 1884. Rodet, Contribution 4 I’Etude Expérimentelle
du Carbon Bactéridien, 1881; Compt. Rend., T. 94, 1882. Roloff, Archiv f.
Wissensch. u. Pract. Thierheilk., Bd. 9, 1883 ; Der Milzbrand: Berlin, 1883.
Schmidt, Deut., Zeitschr. f. Thiermed. u. Verg]. Pathol., 1879. Schrakamp,
Archiv f. Hygiene, Bd. 2, 1884. Semmer, Centralbl. f. d. Med. Wissensch.,
Bd. 18, 1880, 1884; Der Milzbrand und das Milzbrandcontagium, 1882.
Sternberg, Am. Monthly Micr. Journ., 1881. Szpilman, Zeitschr. f. Physiol.
Chemie: Strassburg, Bd. 4, 1880. Toepper, Die Neueren Erfahrungen tiber
d. Aetiologie d. Milzbrands., 1883. Toussaint, Compt. Rend., T. 85, 1877,
1878, 1880; Recherches Expérimentales sur la Maladie Charbonneuse, 1879.
Wachenheim, Etude Expérimentelle sur la Septicité et la Virulence du Sang
‘Charbonneux, 1880.
CHAPTER XV.
QUARTER-EVIL. MALIGNANT CEDEMA. RAG-PICKER’S SEPTIC/MIA
OF GUINEA-PIGS. SEPTICEMIA OF MICE.
QUARTER-EVIL,
_ Arloing, Cornevin and Thomas, Compt. Rend., 1880; Bull. de lAcad. de
Méd., and Revue de Méd., 1881; Du Charbon Bactérien, Charbon Symptoma-
tique, etc., 1883; Revue de Méd., 1884; Chabert’s Disease : Transl. by Dawson
Williams in Micro-parasites and Disease (New Syd. Soc.), 1886. Babés,
Journ. de ’Anatomie, 1884. Bollinger and_Feser, Wochenschr. f. Thierheil-
kunde, 1878. Ehlers, Unters. tib. d. Rauschbrandpilz: Inaug. Diss. Rostock,
1884. Hess, Bericht iiber die entschddigten Rauschbrand u. Milzbrandfalle
im Canton Bern, 1886. Kitt, Jahresber. der K. Thierarzneisch. in Miinchen,
1884. Neelsen and Ehlers, Ber. d. Naturforsch. Ges. zu Rostock, 1884.
Strebel, Schweiz. Archiv f. Thierheilk., 1886.
MALIGNANT CEDEMA.
Brieger and Ehrlich, Berl. Klin. Wochenschr., N. 44, 1882. Chauveau and
Arloing, Archiv Vét., 1884; Bull. Acad. de Méd., 1884. Davaine, Bull. de
YAcad. de Méd., 1862. Gaffky, Mitth. as. d. K. Ges. Amt., 1881. Hesse, W.
and B., Deut. Med. Woch., 1885. Kitasato and Weyl, Zeitschr. f. Hygiene,
Bd. VIII. Kitt, Jahresber. der K. Thierarzneischule in Miinchen, 1884. Koch,
Mitth. aus dem Ges. Amt, L., S. 54, 1881. Lebedeff, Arch. de Phys. Norm. et
Path., 1882. Lustig, Jahresber. d. K. Thierarzneischule zu Hannover, 1884.
Pasteur, Bull. de Acad. de Méd., 1877, 1881. Roger, Compt. Rend. Soc. de Biol.,
1889. Roux and Chamberland, Ann. de l'Institut Pasteur, 1887. Trifaud, Rev.
de Chirurg, T. iii, Van Cott, Centralb. f. Bact., 1891. Verneuil, La Semaine
Méd., 1890.
RAG-PICKER’S SEPTICH/MIA. SEPTICH@MIA OF GUINEA-PIGS. SEPTICH MIA
OF MICE.
Bordoni-Uffreduzzi, Zeitsch. f. Hygiene, 1888. Klein, Centralbl. f. Bac-
teriolog., 1890. Koch, Wundinfectionskrankeit, Trans, New Syd. Soc., 1880.
Paltauf, Wiener Klin. Woch., 1888,
BIBLIOGRAPHY. 651
CHAPTER XVI.
HEMORRHAGIC SEPTICEMIA.
SEPTICEMIA OF BUFFALOES. SEPTIC PLEURO-PNEUMONIA OF CALVES.
SwINE FreveR. SEPTICHMIA OF DEER. SEPTICM@MIA OF RABBITS.
Fow.L CHoLerRa. FowL ENTERITIS. DUCK CHOLERA. GROUSE DISEASE.
Babés, Compt. Rend. de l’Acad. d. 8c., 1883; Arch. de Physiol., 1884.
Barthélémy, Compt. Rend., T. 96, No. 18, 1883, Bunzl-Federn, Centralbl. f.
Bacteriolog., 1891. Camera, Centralbl. f. Bacteriolog., 1891. Cornil, Arch. de
Physiol., Bd. 10, 1882. Cornil and Chantemesse, Le Bulletin Méd., 1887.
Cornil and Toupet, Bull. de la Soc. Nat. d’Acclimation., 1888. Davaine,
Bull, de ?Acad. de Méd., 1879. Eberth and Schimmelbusch, Virchow’s
Archiv, 1889 ; Fortschr. d. Med., Bd. VI. Gaffky, Mitt. aus dem K. G. Amte,
1881. Gamaleia, Centralb. f. Bacteriolog., 1888. Hueppe, Berl. Klin. Woch.,
1886. Ioannés and Mégnin, Journ. d’Acclimatation, 1877. Kitt, Allg. Deut.
Gefliigelzeitung, 1885. Klein, Rep. Med. Off. Local Govt. Board, 1877-78;
Fortschr. der Med., 1888; Centralb. f. Bakteriolog., 1889. Koch, Aetiologie
der Wundinfections R., 1878. Oreste and Armani, Atti de R. Instit, d’Incorrag.
Alle Sci. Nat., Econ e Tech., 1887. Pasteur, Compt. Rend., T. 90, 1886.
Perroncito, Arch. f, Wiss. u. Prakt. Thierheilk., 1879, Petri, Centralbl. f. d.
Med. Wiss., 1885. Rietsch and Jobert, Compt. Rend., 1888. Salmon, Reports
Bureau of Animal Industry, 1886, 1887, 1888. Salmon and Smith, Amer.
Monthly Med. Journ., 1881. Schiitz, Archiv f. Wiss. und Prakt. Thierheilk., 1888.
Semmer, Deut. Zeitschr. f. Theirmed. u. Vergl. Path., 1878. Smith, Journ. f.
Comp. Med. and Surg., 1887; Zeitschr. f. Hygiene, 1891. Smith and Veranus
Moore, Rep. Bureau of Animal Industry, 1895. Ziirn, Die Krankheiten des
Hausgefliigels, 1882.
CHAPTER XVII.
PNEUMONIA. INFECTIOUS PLEURO-PNEUMONIA OF CATTLE.
INFLUENZA.
PNEUMONIA.
Afanassiew, Compt. Rend. Soc. de Biol. Paris, T. 5,1884. De Blasi, Rivista
Internaz. di. Med. e Chir., 1885.: Bruvlant and Verriers, Bull. de l’Acad.
Belge, 1880. Dreschfeld, Fortschr. d. Med., Bd. 3, 389, 1885. Foa and
Bordoni-Uffreduzzi, Deut. Med. Woch., 1886. Frinkel, Verhandl. d. Congr.
f. Innere Med., Fortschr. d. Med, Nov., 1884; Deut. Med. Woch., 1886;
Zeitschr. f. Klin. Med., Bd. x. and xi, 1886. Friedlander, Virchow’s Arcb.,
Bd. 87, 1882; Fortschr. d. Med., Bd. L, 1883; Bd. IL, 1884; Bd. 3, 92, 1885.
Friedlander and Frobenius, Berl. Klin. Woch., 1883. Germain-Sée, Compt.
Rend. Acad. de Sc. Paris, 1884; Des Maladies Spécifiques du Poumon, 1885.
Giles, Brit. Med. J., Vol. IL, 1883. Griffini and Cambria, Centralbl. f. d. Med.
Wiss., 1883. Jaccoud, La France Médicale, 1886. : Jiirgensen, Berl. Klin.
Woch., Bd. 22, 1884. Klein, Centralbl. f. d. Med. Wissensch., 1884. Koranyi
and Babés, Pester Med. Chir. Presse, 1884. Kiihn, Deutsch. Arch. f. Klin.
Med., 1878; Berl. Klin. Woch., Nr. 38, 1881, Lancereaux and Besancon,
652 APPENDICES.
Archiv Gén. de Méd., 1886. Maguire, Brit. Med. Journ., Vol. IL, 1884.
Manfredi, Fortsch. d. Med., 1886. Matray, Wien. Med. Presse, Nr. 23, 1883.
Mendelssohn, Zeitschr. f. Klin. Med, Bd. 7, 1884. Nauwerck, Beitr. zur
Pathol. Anat. von Ziegler, 1884. Neumann, Berl. Klin. Woch., 1885. Paw-
lowsky, Berl. Klin. Woch., 1885. Peterlein, Bericht ii. d. Vet.-Wesen. i. K.
Sachsen, 1885. Pipping, Fortsch. d. Med., Nos. 10 and 14, 1886. Platanow,
Mitth. a. d. Wiirzburg. Med. Klinik, 1885; Ueber die Diagnostische
Bedeutung d. Pneumoniekokken: Inaug.-Diss. Wiirzburg, 1884, Ribbert,
Deut. Med. Woch., Nr. 9, 1885. Salvioli and Zaslein, Centralbl. f. d. Med.
Wissensch., 1883. Arch. pour les Sc. Méd., T. 8., 1884. Schou, Fortschr. der Med.,
Bd. 3, Nr. 15, 1886. Sternberg, Amer. Journ. Med. Sciences, 1885; Journ.
Roy. Micr. Soc., 1886. Talamon, Progr. Médic., 1883. Thost, Deut. Med.
Woch.,, 1886. Weichselbaum, Wien. Med. Woch., 1886. Ziehl, Centralbl. f. d.
Med. Wiss., 1883; Centralbl. f. d. Med. Wiss., 1884.
CEREBRO-SPINAL MENINGITIS.
Bonome, Centralbl. f. Bact. u. Parasitolog., 1V. Foa, Zeitschr. f. Hygiene,
IV. Leichtenstern, Deut. Med. Woch., Nr. 23 u. 31, 1885. Leyden, Centralbl.
f, Klin. Med., Nr. 10, 1883. Weichselbaum, Wien. Klin. Woch., 1888.
PLEURO-PNEUMONIA.
Arloing, Compt. Rend. cix., 1889. Bruce and Loir, Ann. de l'Institut
Pasteur, 1891. Bruylants and Verriers, Bull. de l’Acad. Belg., 1880. Cornil
and Babés, Arch. de Physiol. Norm. et Path., T. 2, 1883. Lustig, Centralb. f.
die Med. Wiss., 1885. Mayrwieser, Woch. f. Thierheilk, u. Viehzucht, 19, 1884.
Pasteur, Recueil de Méd. Vét., 1882. Poels, Fortsch. d. Med., 1886. Poels
and Nolen, Centralbl. f. d. Med. Wiss., Nr. 9, 1884; Fortsch. d. Med., 1886.
Putz, Thier, Med, Vortrige, Bd. 1, 1889. Report on Pleuro-pneumonia and
Tuberculosis, 1888. Schiitz and Steffen, Archiv f. Wiss. und Prakt. Thierheilk.,
Bd. xv. Sussdorff, Deutsche Zeitschr. f. Thiermed. u. Vergl. Pathol., 1879.
INFLUENZA.
Babés, Centralbl. f. Bacteriolog., 1890; Deutsche Med. Woch., 1892. .Bein,
Zeitschr. f. Hygiene, 1890. Bouchard, La Semaine Méd., 1890, Canon, Deutsche
Med. Woch., 1892. Fischel, Prager Med. Woch., 1890; Zeitschr. f. Heilkunde,
1891. Jolles, Wiener Med. Blatt., 1890. Kirchner, Centralbl. f£. Bacteriolog.,
1890; Zeitschr. f. Hygiene, 1890. Kitasato, Deutsche Med. Woch., 1892. Klein,
Brit. Med. Journ., 1892. Klebs, Centralbl. f. Bakteriolog., 1890 ; Deutsche Med.
Woch., 1890. Pfeiffer, Deutsche Med. Woch., 1892. Prudden, New York Med.
Rec., 1890.
CHAPTER XVIII.
ORIENTAL PLAGUE. RELAPSING FEVER. ‘TYPHUS FEVER. ° YELLOW
FEVER.
ORIENTAL PLAGUE.
Aoyama, Mitth. ii. d. Pest. Epidemie im Jahre 1894; in Hong Kong, 1895.
Yersin, Ann, de l'Institut Pasteur, 1894. Yersin, Calmette and Borrel, Ann.
de l'Institut Pasteur, 1895.°.
©
BIBLIOGRAPHY. 653
RELAPSING FEVER.
Albrecht, St. Petersb. Med. Woch., 1879. Carter, Lancet, 1879; Trans.
Internat. Med. Congress, 1881. Cohn, Deut. Med. Woch., 1879. Engel, Berl.
Klin, Woch., 1873. Giinther, Fortschr. d. Med., 1885. Guttmann, Virchow’s
Arch., 1880, Heydenreich, St. Petersb. Med. Woch., 1876 ; Der Parasit des Riick-
falltyphus, 1877. Jaksch, Wien. Med. Woch., Juli, 1880. Koch, Deut. Med.
Woch., 1879. Laptschinsky, Centralbl. f. d. Med. Wiss., Bd. 13, 1875. Manas-
sem, St. Petersb. Med. Woch., No. 18, 1876. Metchnikoff, Virchow’s Archiv,
1877. Moczutowsky, Deutsches Archiv fiir Klin. Med., Bd. xxiv., 1876.
Miihlhaduser, Virchow’s Arch., Bd. 97, 1880. Obermeier, Med. Centralbl., 11;
Berl. Med. Ges.; Berl. Klin. Wochenschr., 1873. Soudakewitch, Ann. de
l'Institut Pasteur, 1891. Weigert, Deut. Med. Woch., 1876.
YELLOW FEVER.
Babés, Compt. Rend., 17 Sept., 1883. Bouley, Compt. Rend., T. 100, p. 1276,
1885. Carmona y Valle, Lecons sur l’Etiol. et la Prophylax. de la Fiévre
Jaune, 1885. Cerecedo, El Siglo Medico, 1886. Domingos Freire, Recherches
sur la Cause de la Fiévre Jaune, 1884; La Fiévre Jaune et ses Inoculations
Préventives, 1896. Domingos Freire and Rebourgeon, Compt. Rend., T. 99,
p. 804, 1884. .
CHAPTER XIX.
SCARLET FEVER AND MEASLES.
SCARLET FVER.
Coze and Feltz, Les Maladies Infectieuses, 1872. Crooke, Lancet, 1883 ;
Fortsch. d. Med., 1885. Crookshank, Report Agric. Dept., 1887. Hahn, Berl.
Klin. Woch., No. 38, 1882. Heubner and Bahrdt, Berl. Klin. Woch., Nr. 44,
1884. Klein, Nature, xxxiv., 1886; Report of the Medical Officer of the
Privy Council, 1887, 1888, 1889. Laure, Lyon Médical, 1886. McKendrick,
Brit. Med, Journal, 1872. Pohl-Pincus, Centralbl. f. d. Med. Wiss., No. 36,
1883. Roth, Miinch. Aerztl. Intelligenzbl., 1883.
MEASLES.
Cornil and Babés, Archiv de Phys., 1883. Keating, Phil. Med. Times, 1882.
CHAPTER XX.
SMALL-POX. CATTLE PLAGUE.
SMALL-POX.
Chauveau, Compt. Rend., 1868. Cohn, Virchow’s Archiv, Bd. 55, 1872.
Copeman, Brit. Med. Journ., 1896; The Practitioner, 1896. Cornil and Babés,
Soc, Méd. des Hép., 1883. Crookshank, History and Path. of Vaccination, 1889.
Haccius, Variola-vaccine, 1892. Hamerink, Ueber die sog. Vac. u. Variola, 188+.
Ischamer, Aerztl. Verein. Steiermark, 1880. Klebs, Arch. f. Exp. Path. u.
Pharmakol., Bd. 10, 1874. Klein, Rep. Med, Off. Loc. Govt, Board, 1893-4.
654 APPENDICES.
Luginbuhl, Verhdl. d, Phys. Med. Ges. in Wiirzburg, 1873, Marchand, Revue
Mycologique, 1882. Pfeiffer, Die Protozoen als Krankheitserreger, 1890;
Behandl. und Prophylax. der Blittern, 1893. Pissin, Berl. Klin. Woch., 1874.
Plaut, Das Organis. Contagium der Schafpocken, 1883. Pohl-Pincus, Vaccina-
tion, 1882. Quist, St. Petersburg Med. Woch., Nr. 46, 1883. Reports, Royal
Vaccination Commission, 1888-96. Ruffer and Plimmer, Brit. Med. Journ.,
1894. Weigert, Ueber Bakterien in der Pockenhaut, 1871; Anat. Beitr. z
Lehre v. d. Pocken, 1874. Wolf, Berl. Klin. Woch., Nr. 4, 1883. Ziilzer, Berl.
Klin. Woch., 1872.
CATTLE PLAGUE.
Crookshank, History and Pathology of Vaccination, 1889. Report of the
Cattle Plague Commission, 1865. Report on Indian Cattle Plague, 1871.
Semmer and Archangelski, Ueber das Rinderpestcontagium und dessen Miti-
gation ; Centralbl. f. d. Med. Wiss., 1883. Simpson, Brit. Med. Journal, 1896.
Smart, Reports on Cattle Plague, Edinburgh, 1865.
CHAPTER XXI.
SHEEP-POX. FOOT AND MOUTH DISEASE.
SHEEP-POX,.
Crookshank, History and Path. of Vaccination, 1889.
Foor anp Moura DISEASE,
Klein, Report Med. Off. Local Govt. Board, 1885. Schottelius, Centralb. f.
Bakteriolog., 1892.
CHAPTER XXII.
HORSE-POX. COW-POX.
HORSE-POXx.
Crookshank, History and Pathology of Vaccination, 1889.
Cow-Pox.
Bucknill, Prov. Med. Journ., 1895. Crookshank, Brit. Med. Journ., 1888;
History and Pathology of Vaccination, 1889 (Vol. II. contains reprints of the
works of Jenner, Pearson, Woodville, Loy, Rogers, Birch, Bousquet, Estlin,
Ceely, Badcock, Auzias-Turenne, Dubreuilh, Layet). Reports of the Royal
Vaccination Commission.
CHAPTER XXIII.
DIPHTHERIA.
Abbot, Bull. Johns Hopkins Univ., 1891, 1893 ; Journ. of Path. and Bact.,
1893. Babés, Virchow’s Archiv. 1890. Behring, Deutsche Med. Woch.
1890. Behring and Kitasato, Deutsche Med. Woch., 1890. Birch-Hirschfeld,
Archiv fiir Heilk., 1872. Brieger and Frankel, Berl. Klin. Woch., 1890.
BIBLIOGRAPHY. 655
Buhl, Zeitschr. fiir Biol., 1867, Cornil, Arch. de Physiol., 1881. Eberth, Med.
Centralbl., XI., Nr. 8, 1873. Emmerich, Compt. Rendus et Mémoires du V.
Congrés Internat. d’ Hygiene, 1884 ; Deut. Med. Woch., 1884. Everett, Med.
and Surg. Reporter, 1881. Forster, Wien. Med. Woch., 1881. Francotte, La
Diphbthérie, 1885. Freidberger, Deut. Zeitschr. fiir Thiermed. u. Vergl. Pathol.,
1879. Fiirbringer, Virch. Arch., Bd. 91, 1883. Gerhardt and Klebs, Verhandl.
d. Congresses f. Inn. Med., 1882, 1883. Heubner, Die Experimentelle Diph-
therie, 1883. Hueter and Tommasi, Centralbl. f. d. Med, Wiss., 1868. Klebs,
Arch. f. Exp. Pathol. Bd. 4, 1875. Klein, Report Med. Dept. Local Govt.
Board, 1889. Letzerich, Berl. Klin. Woch., xi., 1874; Virch. Arch., Bd. 53,
1872; Bd. 68, 1876. Loffler, Mittheil. a. d. Kais. Ges. Amt, Bd, IT., 1884 ;
Microparasites and Disease (New Syd. Soc.), 1886 ; Centralbl. f. Bacteriolog.,
1887, 1890; Deutsche Med. Woch., 1890. Lumner, Aerzil. Int. Bl, No. 31,
1881. Martin, Rep. Med. Off. Loc. Gov. Board, 1890. Neumayer, Neue
Thesen zu Diphtheritisfrage, 1880. Nicati, Compt. Rend., T. 88, 1879. Oertel,.
Deut, Arch. f. Klin, Med., Bd. 8, 1871; Zur Aetiologie der Infectionskrank-
heiten, 1881. Park, New York Med. Record, 1892, 1893. Prudden, Amer.
Journal of Med. Sci., 1889 ; New York Med. Rec., 1891. Rivolta, Giornale de
Anat. Fisiol. e Patol. delli Anim., 1884. Roux and Yersin, Ann. de l'Institut
Pasteur, 1888, 1889, 1890, Salisbury, Gaillard’s Med. Journ.: New York, 1882.
Talamon, Bull. de la Soc. Anat. de Paris, T. 56, 1881. Welch, Amer, Journal
of the Med. Sci., 1894. Welch and Abbott, Bull. Johns Hopkins Univ., 1891.
Wood and Formad, Bull. Nat. Board of Health, Wash. and Med. Times and
Gazette, 1882. Zahn, Beitriige zur Pathol. u. Histol. der Diphtherie, 1878.
Zarniko, Centralbl. f. Bact., 1889.
CHAPTER XXIV.
TYPHOID FEVER.
Almquist, Typhoidfeberus-Bakterie, 1882. Beumer and Peiper, Zeitschr. f.
Hygiene, 1886. Birch-Hirschfeld, Zeitschr. f. Epidemiologie, I., 1874. Boens,
Acad. Roy. de Méd. de Belgique: Bull. 3 Sér., T. 17, 1883. Brautlecht,
Virchow’s Arch., Bd. 84, 1881. Coats, Brit. Med. Journ., 1882. Crooke, Brit.
Med. Journ., 1882. Eberth, Arch. f. Pathol. Anat., Bd. 8), 1880; Volkmann,
Klin. Vortriige, 1883. Eppinger, Beitr. zur Pathol. Anatomie aus d. Patholog,
Institut. Prag., 1880. Feltz, Compt. Rend., T. 85, 1877. Fischel, Beitr. zur
Pathol. Anat. aus d. Pathol.-Anat. Inst. Prag., 1880. Fraenkel and Simmonds,
Die Aetiologische Bedeutung des Typhus-Bacillus, 1886. Gaffky, Mitth, a, d.
Ges. Amt, Bd. IL, 1884. Klebs, Arch. f. Exper. Pathol. u. Pharmakol., 1880.
Klein, Med. Centralbl., xii., 1874, Letzerich, Virchow’s Archiv, Bd. 68, 1876 ;
Archiv f. Exper. Pathol., Bd. 9, 1878; Ba. 10, 1881; Experimentelle Unter-
suchungen tiber die Aetiologie des Typhus Abdominalis : Leipzig, 1883. Luca-
tello, Bollet. d. R. Accad, Med. di Genova, 1886. Maraghano, Centralbl. f. d.
Med. Wiss., Bd. 15, 1882. Meisels, Wien. Med. Woch., 1886. Meyer, Unters.
jiber den Bacillus des Abdominaltyphus: Inaug. Diss., 1881. Michael, Fortsch.
d. Med., 1885. Neuhauss, Berl. Klin. Woch., 1886. Pfeiffer, Deut. Med.
Woch., Nr. 29, 1885. Rappin, Contrib. 4 l’Etude des Bact. de la Bouche,
4 Etat Normal et dans la Fievre Typhoide, 1881. Seitz, Bakteriolog. Studien
z. Typhusitiologie, 1886. Sirotinin, Zeitsch. f. Hygiene, 1886. Tayon, Compt.
Rend., T. 99, p. 331, 1884. Tizzoni, Studi. di Pat. Sperim. sulla Gen. d. Tifo.:
Milan, 1880. Wernich, Zeitschy. f. Klin, Med., Bd. 6, 1882.
656 APPENDICES.
CHAPTER XXvV.
SWINE-TYPHOID.
Klein, Rep. of the Med. Offic. of the Privy Council, 1877-78 ; Virchow’s
Archiv, 1884. M‘Fadyean, Journ. of Comp. Path. and Therapeutics, 1895.
Report of a Conference on Swine-fever: Board of Agriculture, 1896. Rietsch,
Jobert and Martinaud, Compt. Rend., T. 106. Selander, Centr. f. Bakt. u.
Parasitenk., 1888 ; Ann de l'Institut Pasteur, 1890. Smith and Veranus Moore,
Bull. of Bureau of Animal Industry U.S., 1894. Welch and Clement, Trans.
Internat. Vet. Congress of Amer., 1893.
CHAPTER XXVI.
SWINE-MEASLES. DISTEMPER IN DOGS. EPIDEMIC DISEASE OF
FERRETS. EPIDEMIC DISEASE OF MICE.
SWINE-MBASLES.
Cornil and Babés, Arch. de Physiol. 1883. Detmers, Science, 1881.
Eggeling, Fortschr. d. Med., 1883. Léffler, Arbeiten aus dem Kaiser]. Gesund-
heits Amt, Bd. 1., 1885. Lydtin and Schottelius, Der Rothlauf der Schweine :
Wiesbaden, 1885. Pasteur, Compt. Rend., T. 95, 1882. Pasteur and Thuillier,
Bull. de Acad. de Méd. de Paris: Compt. Rend., T. 97, 1883. Salmon,
Report Depart. Agricul. Washington, 1881, 1884. Schiitz, Ueber den Rothlauf
der Schweine und die Impfung desselben, 1885.
CHAPTER XXVII,
ASIATIC CHOLERA. CHOLERA NOSTRAS. CHOLERAIC DIARRHG&A
FROM MEAT-POISONING. DYSENTERY. CHOLERAIC DIARRHGA
OF FOWLS.
CHOLERA.
Ali-Cohen, Fortschr. der Med., 1887, 1888. Almquist, Zeitschr. f. Hygiene,
1887. Babés, Virchow’s Archiv, 1885. Bianchi, Lancet., 1884. Bitter, Archiv f.
Hygiene, 1886. Bochefontaine, Expér. pour servir 4 l’Etude des Phénoménes
déterminés chez Homme par l’Ingestion Stomacale du Liquide Diarrhéique
du Choléra: Compt. Rend., 1884. Brieger, Deutsche Med. Woch., 1887; Berl.
Klin. Woch., 1887; Virchow’s Archiv, 1887. Brunetti, Fatti Considerazioni
Conclusioni sul Coléra, 1885, Biichner, Archiv f. Hygiene, 1885; Miinchn. Aerztl.
Intelligenzbl., 8. 549, 1884. Btichner and Emmerich, Miinch. Med. Woch., 1885.
Bujwid, Zeitschr. f. Hygiene, Centralbl. f. Bacteriolog., 1888. Cameron, Brit.
Med. Journ., 1884. Cattani, Deut. Med. Woch. 1886. Carter, Lancet, 1884.
Cheyne, Brit. Med. Journ., 1885. Crookshank, Lancet, 1885. Cunningham,
Scientific Memoirs of Med. Off. of Indian Army, 1885, 1891. Deneke, Deut.
Med. Woch., Nr. 3, 1885. Doyen, Soc. de Biol. de Paris, 1884. Drasche,
Allg, Wien. Med. Zeit., 1885. Dunham, Zeitsch. f. Hygiene, 1887. Emmerich,
Deut, Med. Woch., Nr. 50, 1884, Ermengem, Deut. Med. Woch., Ny. 29, 1885;
BIBLIOGRAPHY. 657
Recherches sur le Microbe du Choléra Asiatique, 1885. Ferran, Compt. Rend.,
T. 100, p. 959, 1885. Finkler and Prior, Naturforscherversammlung Magde-
burg, 1884; Deut. Med. Woch., Nr. 36, 1884; Ergiinzungshefte zum Centralbl.
f, Allg.. Gesundheitspflege, Bd. I, Heft 5 u. 6, 1885. Fliigge, Deut. Med.
Woch., Nr. 2, 1885. Gaffky, Arb., aus d. K. Gesundheitsamte, 1887. Gamaleia,
Compt. Rend., 1888; Compt. Rend. Soc. de Biolog., 1889. Gibier and van
‘Ermengem, Compt. Rend., T. 101, 1885. Haffkine,. Brit, Med. Journ., 1895.
Héricourt, Compt. Rend., 1885. Hueppe, Berl. Klin. Woch., 1887; Deutsche
Med. Woch., 1887; Compt. Rend., 1889; Prag. Med. Woch., 1889. Hunter,
Brit. Med. Journ., 1884. Johne,*Deutsche Zeitschr. f. Thiermed., Bd. XI, 1885.
Kitasato, Zeitschrift f. Hygiene, 1488, 1889. Klebs, Ueber Ciolera Asiatica,
1885. Klein, Brit. Med. Journ, and Proc. Roy. Soc. London, No. 38, 1885;
Bacteria of Asiatic Cholera, 1889. Klein and Gibbes, An Inquiry into the
Etiology of Asiatic Cholera: Bluebook, 1885. Koch, Deut. Med. Woch., 1884 ;
Etiology of Cholera: Berlin Cholera Conference: Translated by Laycock,
in Microparasites and Disease (New Syd. Soc.), 1885. Lewis, Med. Times
and Gazette, 1884. Lustig, Centralbl. f. die Med. Wiss., 1887; Zeitschrift f.
Hygiene, 1887. Macnamara, Brit. Med. Journ., 1884, Miller, Deut. Med.
Woch., Nr. 9, 1885. Neuhaus, Centralbl. f. Bacteriolog., 1889. Nicati and
Rietsch, Arch. de Physiol. 1885: Revue de Médecin, T. 5, 1885; Revue
dHygiéne, 1885. Petri, Centralbl. f. Bacteriolog., 1889. Pettenkofer, Lancet,
1886. Pfeiffer, Corresp.-Bl. des Allgem. Aerztl. Ver. in Thiiringen, Nr. 9, 1884;
Deut. Med. Woch., 1886-88. Pfuhl, Zeitschr. f. Hygiene, 1889. Roy, Brown-
and Sherrington, Proc. Roy. Soc., Vol. xli. Salkowski, Virchow’s Arcuiv, 1837.
Schottelius, Deut. Med. Woch., Nr. 14, 1885, Strauss, Roux, Thuillier and
Nocard, Compt. Rend. Soc. de Biol. T. 4, 1883. Tizzoni and Cattani,
Centralb. f, die Med. Wiss., 1886, 1887. Vineenzi, Deutsche Med. Woch., 1887.
Wassiljew, Zeitschr. f. Hygiene, 1847. Weisser, Zeitschrift f. Hygiene, 1886.
Weisser and Frank, Zeitschr. f. Hygiene, 1886. Zaslein, Deutsche Med.
Zeitung, 1887-88.
CHAPTER XXVIII.
‘TUBERCULOSIS.
Albrecht, Arch. f. Kinderheilk., Bd. 5, 1884. Andrew, Lancet, 1884.
Arloing, Compt. Rend., 1884. Aufrecht, Centralbl. fiir d. Med. Wiss., 1882, 1883.
Babés, Compt. Rendus, 1883 ; Centralbl. f. d. Mei. Wissensch., 1883. Babés
and Cornil, Journ. de /’Anat. et de la Physiol. Norm. et Pathol. 1884, Balogh,
Wien. Med. Woch., 1882. Baumgarten, Berl. Klin. Wo-h., Bd. 17, 1879;
Centralbl. f. d. Med. Wiss., Bd. 19, 1881 ; Bd, 20, 1882; Bd. 21, 1883; Bd. 22,
1884; Deut. Med. Woch., Bd. 8, 1882; Zeitscbr. f. Klin. Med., Bad. 6, 1883,
1886. Biedert, Virchow’s Archiv, Bd. 98, 1884 ; Berl. Klin. Woch., 1885. Black,
Lancet, 1886. Bock, Virchow’s Archiv, Bd. 91, 1883, Bollinger, Centralbl. f.
d. Med. Wiss., Bd. 21, 8. 600, 1883 ; Miinch. Aerztl. : Intelligenzbl., Nr. 16,
1883. ,Bouley, La Nature Vivante de 1h Contagion, Contagiosite de la Tuber-
84. Brouilly, Rev. de Chir., T. 3., 1883. Celli and Guarneri,
Arch, pour les Sciences Médic, 1883. Cheyne, Brit. Med. Journ., Vol. I., 1883 ;
Practitioner, Vol. XXX., 1883. Chiari, Wien. Med. Presse, 1883. Cochet,
Compt. Rend. Soc. de Biol. : Paris, I. 5, 1883. Cohnheim, Uebertragbarkeit
der Tuberculose: Berlin, 1877. Cornil and Leloir, Arch. de Physiol. Norm. et
42
culose: Paris, 18
658 _APPENDICES.
Pathol, 1884, Cramer, Sitzungsber. d. Phys. Med. Soc. zu Erlangen, 1883,
Creighton, Trans. Path. Soc., 1882 ; Brit. Med. Journ., 1885; Lancet, 1885.
Crookshank, Rep. Agric. Dept., 1888 ; Proc. Phys. Soc., 1890 ; Trans. Path. Soc.,
1891. Damsch, Deut. Med. Woch., Nr. 17, 1883. Déjérine, Rev. de Méd.: Paris,
T. 4,1884. Demme, Jahresber. d. Jennerschen Kinderspitals: Bern, 1883. Dett-
weiler, Berl. Klin, Woch., Bd. 21, 1880, Dieulafoy and Krishaber, Arch. de
Physiol. Norm. et Path., T. I., 1883. Doutrelepont, Vierteljahrschr. f. Dermato-
logie u, Syphilis, 1884; Deut. Med. Woch., Nr. 7, 1885. Ehrlich, Deut. Med.
Woch., 1882, 1883. Ermengem, Ann. de la Soc. Belge de Microscopie, 1882.
Ewart, Lancet, 1882. Formad, The Bacillus Tuberculosis: The Philad.
. Medical Times, 1882. Frantzel and Palmers, Berl. Klin. Woch., 1882, 1883,
Fiitterer, Virch. Arch., Bd. 100, Heft 2, 1885. Gaffky, Mitth. a. d. Kaiserl.
Gesundheitsamte, Bd. II., 1884. Giacomi, Fortschr. d. Med., Bd. L, 8. 145,
1883. Gibbes, Lancet, 1883. Goldenblum, Vrach., Nos. I. and XI., 1886.
Green, Brit. Med Journ., 1883 ; Lancet, 1887. Harries and Campbell, Lupus:
London, 1886. Harris, St. Barthol. Hosp. Reports, 1885. Heron, Lancet,
1883. Hiller, Deut, Med. Woch., Bd. 8, 1882. Johne, Die Geschichte der
Tuberculose, 1883; Ber. iib. d. Veterindrwesen im KG6nigr: Sachsen, 1883;
Fortschr. d. Med., Bd. 3, 198, 1885. Karg, Centralbl. f. Chir., 1885. Kirstein,
Deut. Med. Woch., 1886. Klebs, Virchow’s Arch., Bd. 44, 1868; Arch. f. Exp.
Pathol. u. Pharmakol., Bd. I., 1873 ; Bd. 17, 1883. Koch, Die Aetiologie der
Tuberculose: Berl. Klin. Woch., 1882; Deut. Med. Woch., Nr. 10, 1883;
Mittheilungen aus dem Kais. Ges. Amt, Bd. II., 1884. Kundrat, Wien. Med,
Presse, 1883. Kiissner, Deut. Med. Woch., Nr. 36, 1883. Landouzy and
Martin, Rev. de Méd., T. 3, 1883. Leube, Sitzungsber. der Phys.-Med. Soc. zu
Erlangen, 1883. Levinsky, Deut. Med. Woch., Bd. 9., 1883. Leyden, Zeitschr.
f. Klin. Med., VIII, 1884. Lichtheim, Fortschr. d. Med., Bd. I., 1883. Lustig,
Wien. Med. Woch., Nr. 48, 1884. Lydtin, Badische Thierarztl. Mittheil, 1883.
Malassez and Vignal, Compt. Rend., T. 97, 1883 ; Compt. Rend., T. 99, p. 200,
1884. Marchand, Deut. Med. Woch., Nr. 15, 1883. Max-Bender, Deut. Med..
Woch., 1886. Meisels, Wien. Med. Woch., 1883. Middeldorpf, Fortsch. d.
Med., 1886. Miiller, Centralbl. f. Chir., 3; 1884, 1886. Nauwerck, Deut.
Med. Woch., 1883. Obrzut, Deut. Med. Wocb., Nr. 12, -1885. Pfeiffer,
Berlin. Klin. Woch., Bd. 21, 1883. Piitz, Ueber die Beziehungen der
Tuberculose des Menschen zu der Thiere, 1883. Raymond, Arch. Gén. de
Méd., T. 11, 1883. Ribbert, Deut. Med. Woch., 1883, 1885. Rindfleisch, Phys.
Med. Ges. zu Wiirzburg, Nr. 8, 1882. Rosenstein, Centralbl. f. d. Med. Wiss.,
1883. Schill and Fischer, Mitth. a. d. Kaiserl. Ges. Amt, Bd. IL, 1884.
Schlegtendal, Fortschr. d..Med., Bd. I., 1883. Schottelius, Virchow’s Archiv.
Bd. 91, 1883. Schuchardt and Krause, Fortschr. d. Med. 1883. Smith, Bristol
Med. Chir. Journ., 1883. Somari and Brugnatelli, Redii R. Instit. Lombardo,
1883. Spina, Casopis Lekaru Ceskych, Nr. 4, 1885 ; Studien tiber Tuberculose :
Wien., 1883. Sticker, Centralbl. f. Klin. Med., 1885. Strassmann, Virchow’s
Archiv, Bd. 96, 1884. Sutton, Trans. Path.’ Soc. London, Vol. XXXV., 1884.
Toussaint, Compt. Rend., T.93,18%1. Tscherning, Fortschr. d. Med., Bd. 3, 65,
1885. Veraguth, Arch. f. Exp. Path. u. Pharmakol., Bd. 16, 1883, Vignal,
Compt. Rend. Soc. de Biol., T. 5, 1883. Villemin, Etude sur la Tuberculose,
1868, Voltolini, Deut. Med. Woch., Nr. 31. Wahl, Deut. Med. Woch., Nr. 46,
1882, Weichselbaum, Wien. Med. Jahrb., 1883. Weigert, Deut. Med. Wocb.,
Nr. 24ff. 1883. Wesener, Fiitterungstuberculose, 1885. West, Lancet, Vol. I.,
1883 ; Trans. Path. Soc., 1883. Williams, Lancet, 1883; Journ. Roy. Mier.
Soc., 1884. Ziehl, Deut. Med. Woch., Nr. 5. 1883.
BIBLIOGRAPHY. si 659
CHAPTER XXIX.
LEPROSY. SYPHILIS. RHINOSCLEROMA. TRACHOMA.
LEpPRosy.
Arning, Virchow’s Archiv, Bd. 97, 1884. Babés, Compt. Rend., 1883. Babés
and Kalindero, La Semaine Med., 1891. Baumgarten, C'entralbl. £ Bact.,
1887; Berl. Klin. Woch., 1889. Beaven-Rake, Trans. Path. Soc., 1887 ;
Reports Leper Asylum, Trinidad. Bordoni-Uffreduzzi, Zeitschr. f. Hygiene
1887; Berl. Klin. Woch., 1888. Campava, la Reforma Med., 1889, 1891.
Cornil and Suchard, Ann. de Dermat. et Syph., 1881. Creighton, History
of Epidemics in Great Britain, 1894. Damsch, Virch. Arch., Bd. 92:
Centralbl. f. d. Med. Wissensch.. Bd. 21, 1883. Gaucher and Hillairet,
Progrés Méd., 1880. Guttmann, Berl. Klin. Woch., 1885. Hansen, Virchow’s
Archiv, 1880, 1882, 1886. Hillis, Trans. Path. Soc., 1883. Hills, On Leprosy
in British Guiana, 18%1. Kaposi, Wiener Med. Woch., 1X83, Kébner,
Virchow’s Arch., 1882 ; Leloir, Ann. de Dermat. et de Syph., 1887. Lubimoff,
Centralbl. f. Bacteriolog., 1888. Melcher and Ortmann, Berl. Klin. Woch.
1885, 1886. Moretti, Tl Primo Caso di Lebbra nelle Marcho Confermato dalla
Presenza del Bacillus Lepr, 1883, Miiller, Deut. Archiv f. Klin. Med., Bd.
34, 1883. Neisser, Breslauer Aerztl. Zeitschr., 1879; Jahresber. d. Schles
Ges. fiir Vaterl. Cultur., 1879 ; Virchow’s Archiv, Bd. 84, 1881, 1886. Newman,
Leprosy as an Endemic Disease in the British Islands, 1895. Report, Leprosy
Commission, 1893. Steven, Brit. Med. Journ., 1885. Thin, Brit. Med.
Journ., 1884. Touton, Fortsch. d. Med.. 1886. Unna, Deut. Med. Woch..
Nr. 32, 1885, 1886 ; Virchow’s Archiv, 1886. Unna and Lutz, Dermatologische
Studien, Heft I., 186. Vidal, La Lepre et son Traitement, 1884. Virchow,
Berl. Klin. Woch., N. 12, 1885. Vossius, Ber. iiber d. Ophthalmologen Con-
gress in Heidelberg, 1881. Wesener, Central. f. Bacteriolog., 1887 ; Munch.
Med. Woch.,-1887.
SYPHILIS.
Alvarez and Tavel, Bull. de l’Acad. de Méd. et Archiv de Phys. Norm. et
Path,, 1885. Aufrecht, Centralbl. f. d. Med. Wissensch., Bd. 19, 1881.
Bienstock, Fortsch. d. Med., 1886. Birch-Hirschfeld, Centralbl. f. d. Med.
Wissensch., Nrs. 33, 34, 1882. Disse and Taguchi, Deut. Med. Woch., 1886.
Doutrelepont and Schiitz, Deut. Med. Woch., Nr. 19, 1885. Eve and Lingard,
Brit. Med. Journ., 1886. De Giacomi, Correspondenzbl. f. Schweizer Aerzte,
Bd. 15, 1885. Gottstein, Fortschr. d. Med.. Bd. 3, S. 543, 1885. Heyden,
Préservation de la Svphilis, etc.: Traduit par Roberts, 1883, Kassowitz
and Hochsinger, Wien. Med. Blatter, 1886. Klebs, Arch. f. Exp. Pathol.
Bd. 10, 1879. Klemperer, Deut. Med. Woch., 1885. Koniger, Deut. Med.
Woch., 8. 816, 1884. Letnik, Wien. Med. Wochenschr., 1883. Lostorfer, Arch.
f. Dermat. u. Syph., 172. Lustgarten, Wien. Med. Woch., Nr. 47, 1884; Die
Syphilisbacillen, 1885. Martineau and Hamonic, Compt. Rend., Pp. 443, 1882,
Morison, Maryland Med. Journ., 1882 ; Ibid. : Baltimore, 1883; Wiener Med,
Wochenschr., 1843. Peschel, Centralbl. f. Augenheilk., 1882. Petrone, Gaz.
Medica Ital., 1884, Torney and Marcus, Compt. Rend., p. 472, 1st 4.
RHINOSCLEROMA.
Cornil and £7var2z, Acad. de
Cornil, B. de Ja Soc. Anatom., 15 Fev., 185. :
Davies, Brit. Med. Journ,
Méd. et Archiv de Phys. Norm. et Path., 1885,
1886. Paltauf and Eiselsberg, Fortsch. d. Med,, 1888.
660 APPENDICES,
CHAPTER XXX.
ACTINOMYCOSIS AND MADURA DISEASE,
ACTINOMYCOSIS.
Acland, Brit. Med. Journ. and Trans. Path. Soc., 1886 ; and Allbut’s System of
Med., 1896. Bang, Tidskrift far Veterinaerer, 1883. Baumgarten, Berl. Klin.
Woch., 1885. Bollinger, Centralbl. f. d. Med. Wiss., 1877. Bostrém, Verh, d.
Congr. f. Inn. Med. Wiesbaden, 1888. Chiari, Prager Med. Woch., Nr. 10, 1884.
Crookshank, Report of the Agric. Dept. of the Privy Council, 1888; Trans.
Roy. Med. and Chirurg. Soc., 1889. Firket, Rev. de Méd., 1884. Fleming,
Actinomycosis, 1883. Gannet, Boston Med. and Surg. Journ., 1882. .Hertwig,
Archiv f. Wiss. u. Prakt. Thierheilk., 1886. Hink, Centralbl. f. d. Med. Wiss.,
1882. Israél, Archiy f. Klin. Chir., 1868 ; Virchow’s Arch., Bd. 74, 1878 ; Bd. 78,
1879; Bd. 96, 1884; Kenntniss der Actinomykose des Menschen: Klinische
Beitrage, 1885. Johne, Deutsche Zeitschr. f. Thiermed., 1881 ; Bericht ii. d.
Veter.-Wesen i. K. Sachsen, 1885. Karsten, Deut. Med. Woch., 1884. Mag-
nussen, Beitriige zur Diagnostik u, Casuistik der Actinomykose : Diss. Kiel.,
1885. Mitteldorpf, Deut. Med. Woch., 1884. Maleolm Morris, Brit. Med. Journ.,
1896. Murphy, New York Med. Journ., 1885. O’Neill, Lancet, 1886. Pflug,
Centralbl. f. d. Med. Wiss., 1882. Ponfick, Breslauer Aerztl. Zeitschr., 1885 ;
Die Actinomykose: Berlin, 1887. Pusch, Arch. f. Wiss. u. Pr. Thierheilk.,
1883. Ransome, Brit. Med. Journ., 1896. Report of the Board of Live-Stock
Com. for the State of Illinois, 1890, Roser, Deut. Med. Wocb., 1886. Soltmann,
Breslauer Aerztl, Zeitschr., 1485. Treves, Lancet, 1884. Zemann, Wien. Med.
Jahrb., 8. 477, 1883.
Mapvura DISEASE,
Boyce and Surveyor, Proc. Roy. Soc., 1893 ; Trans. Roy. Soc., 1894; Brit.
Med. Journ., 1894. Hewlett, Lancet, 1894. Surveyor, Brit. Med. Journ., 1892.
Vincent, Ann, de l'Institut Pasteur, 1894.
CHAPTER XXXI.
GLANDERS.
Babés, Acad. de. Med., 1888. Baumgarten, Centralbl. f. Bacteriolog., 1888.
Bouchard, Capitan and Charrin, Bull. de lAcad. d, Sc., Nr. 51, 1882.
Cadeac and Malet, Progrés Méd., 1886, 1887; Rec. de Méd. Vt., 1886 ; Oester.
Monatschr. f, Thierheilk., 1888. Fréhner, Rep. d. Thierheilk., 1883, Griinwald,
Oesterr. Monatsschr. fiir Thierheilk., Nr. 4, 1884, Hunting, Vet. Record.,
1896. Israél, Berl. Klin. Woch., Nr. 11., 1883. Kitt, Jahresber. d. Miinchen.
Vhierarzneisch, 1884. Kranzfeldt, Centralb. f. Bacteriolog., 1887. Liffler,
Arbeit. a, d. K. Gesundh. Amt., 1886. Loffler and Schutz, Deut Med. Woch.,
Nr. 52, 1882, ‘Molkentin, Zur Sicherstellung der Diagnose von Rotz: Inaug.-
Diss., 1883. Raskin, Zeit. f. Wiss. Mikroscop., 1837. Salmon, Reports Bureau
of Animal Industry, 1887, 1888. Smith, Journal of Comp. Med. and Veterin.
Archives, 1890. Struek, Deut. Med. Woch., Nos. 51 u. 52, 1883. Vulpian and
Bouley, Bull. de 1’Acad. de Méd., 1883. Wassilieff, Deut. Med. Woch., Nr. 11.,
1883, Weichselbaum, Wiener Med. Woch., 21-24, 1884.
BIBLIOGRAPHY. 661
CHAPTER XXXII.
TETANUS. RABIES, LOUPING-ILL.
TETANUS.
Carle and Rattone, Studio Sperimentale sull’ Etiologia del Tetano. Giorn.
della R, Acad, di Medicina di Torino, 1884. Hewlett, Brit. Med, Journ,, 1895.
Martin, Rep. Med. Off. Local Govt. Board 1895. . Nicolaier, Deut. Med.
Woch., Nr. 52, 1884. Rosenbach, Archiv f. Klin. Chir., 1886. Roux and
Vaillard, Ann. de l'Institut Pasteur, 1893. Vogel, Deut. Med. Woch., Nr. 31,
1884,
RABIES.
Babés, Les Bactéries, 1886. Bauer, Minch. Med. Woch., 1886. Bert,
Compt. Rend., 1882. Colin, Bull. Acad. de Méd. Paris, T. 10, 1881. Doléris,
Gaz. Méd. de Paris, T. 3: Tribune Méd. Paris, T. 14, 1881. Dowdeswell,
Journ, Roy. Micro. Soc. and Lancet, 1886. Fol, Acad. des Sciences, 1884.
Frisch, Wien. Med. Woch., 1886. Gibier, Compt. Rend., 1883. Kerr, Brit.
Med, Journ., 1886. Pasteur, Compt. Rend., 1881, 1884, 1886 ; Ann. de Méd.
Vétérin, 1884. Pasteur, Chamberland, Roux and Thuiller, Compt. Rend., 1882.
Percheron, La Rage et les Expériences de M. Pasteur, 1884. Report of the
English Hydrophobia Com. Vignal, Brit. Med. Journ., 1886.
LOUPING-ILL.
Klein, Journ. of Royal Agric. Soc., 1895. M‘Fadyean, Journ. Royal Agric.
Soc., 1895, and Journ. of Comp. Path. and Bacteriology, 1895.
CHAPTER XXXIII.
FOOT-ROT.
Brown, Journ. Royal Agric. Soc., 1892. Nott, Journ. Royal Agric, Soc.,
1890.
CHAPTER XXXIV.
FOUL-BROOD. INFECTIOUS DISEASE OF BEES IN ITALY. PEBRINE.
FLACHERIE. INFECTIOUS DISEASE OF CATERPILLARS.
Béchamp, Compt. Rend., 1867. Cheshire and Cheyne, Journ. of Roy. Micros.
Society. Cowan, Journ. Royal Agric. Soc., i892. Forbes, Bull. Illin, State
Lab. of Nat. Hist., 1886. Klamann, Bienenwirthschaft. Centralbl., 1888.
Pasteur, Etudes des maladies des ver a soie, 1870.
CHAPTER XXXV. is
CLASSIFICATION AND DESCRIPTION OF SPECIES.
Acosta and Rossi, Centralbl. f. Bact., 1893. Adametz, Die Bakt. der Nutz.
u. Trinkwiisser, 1888 ; Landwirthschaft. Jarhb., 1890. Afanassiew, St. Peters-
burg Med. Wech., 1887, Ali-Cohen, Centr. f. Bact., Bd. VI., 1889. Almquist,
662 APPENDICES.
Zeitschr. f. Hygiene, Bd. X., 1891. Alvarez, Compt. Rend., T. cv., 1887.
Arloing, Compt. Rend., cvi. and cvii, Arthur, Proc. Acad. Nat. Sci. Phil., 1886.
Babés, Bakt. Untersuch. ii, Sept. Proz. des Kindesalters, 1889 ; Virch. Archiv,
Bd. exy.,1889 ; Centralb. f. Bacteriolog., Bd. IX., 1891 ; Progrés Med. Roumain.
Babés and Oprescu, Ann. de VInstitut Pasteur, Vol. v., 1891. Baginsky,
Deutsche Med. Woch., 188%. Banti, Giornale Medico, 1888. Beyerinck, Bot.
Zeitung, Vol. xlix., 1891. Bienstock, Zeitschr. f. Klin. Med., VIII. Billet,
Compt. Rend., T. 100., 1885. Biondi, Zeitschrift. f. Hygiene, Bd. IL., 1887.
Bizzozero, Virchow’s Archiv, xcviii. Bolton, Amer. Journ. Med. Sci., 1892 ;
Zeitsch. £. Hygiene, Bd. I., 1886. Bonome, Archiv per le Sci. Med., Vol. xiii.,
1890, Booker, Transac. Ninth Internat. Med. Congress, Vol. III. Bordoni-
Uffreduzzi, Fortsch. der Med., 1886; Zeitschr. f. Hygiene, Bd. III., 1888.
Botkin, Zeitschr. f. Hygiene, Bd. XI., 1892. Bouchard, Compt. Rend.,
T. cviii., 1889. Bovet, Ann. de Micrographie, Vol. iii, 1891. Brannan and
Cheeseman, New York Medical Record, 1892. Brefeld, Ges. Nat. Freunde,
1878. Breunig, Bakt, Untersuch. d. Trinkwiassers der Stadt Kiel, 1888.-
Brieger, Berl. Klin. Woch, 1884. Brouardel and Boutmy, Compt. Rend., T. 92,
p. 1056. Bujwid, Centralb. f. Bacteriolog., 1893. Bumm, Der Mikr. der
Gon. Schleimk., 1887. Burri, U. eim. « Zuleck. d. Artchar auz Bact. Unter-
such. a. Rhein W. Isol. Bact., 1893, Burrill, Proc. Amer. Assoc. Advance. Sci.,
1880. Caldo, Bull. de la Soc. d’Anatom. de Paris, 1887. Canon and Pielicke,
Berl. Klin. Woch., 1892. Caspary, Schriften der Physik. Oekon. Ges, zu
Konigsberg, Bd. 15,1874. Cazal and Vaillard, Ann. de |’ Instit. Pasteur, Vol. v.,
1891. Charrin, La Maladie Pyocyanique, 1889. Cheyne, Brit. Med. Journ.,
1886. Cienkowski, Die Gallertbild. des Zuckerrubensaftes, 1878; Zur Moyr-
phol. des Bakter., 1876. Clado, Bull. de la Soc. d’Anat. de Paris, 1887.
Classen, Centr. f. Bakt., Bd. VIL, 1890. Cohn, Max Schultz’s Archiv, Bd. III. ;
Ueber zwei Neue Beggiatoen: Hedwigia, 1865; Beitr. z. Biol. d. Pflanzen,
Bd. I., 1872. Conn, Centralbl. f. Bakteriol, Bd. IX., 1891. Dallinger,
Journ. of the Roy. Micro. Soc. London, 1878. Dallinger and Drysdale,
Monthly Microsc. Journ., 1875. Dantec, Ann. de l'Institut Pasteur, 1891.
De Bary, Vergl. Morph. und Biol. der Pilze, 1884. Demme, Fortschr. der
Med., 1888; Verhandl. der V. Congress fiir in Med. in Wiesbaden, 1886.
Deneke, Deutsch. Med. Woch., 1885. Dickerhoff and Grawitz, Virchow’s
Archiv, Bd. cii., 1885. Dowdeswell, Ann. de Micrographie, II. berth,
Virchow’s Archiv, Bd. 13, 1858. Ehrenberg, Verhandl. der Berl. Acad., 1839.
Eidam, Cohn’s Beitr. zur. Biol. d. Pflanzen, 1875. Eisenberg, Bacteriolog.
Diagnostik, 1891. Emmerich, Deutsche Med. Woch., 1884. Engelmann,
Pfliiger’s Arch. f. d. Ges. Physiol., Bd. 30, 8. 95, 1882, Engler, Bericht. d.
Kommission z. Erfolsch. Deutsch. Meere, 1881. Ernst, Zeitschr. f. Hygiene,
Il. Escherich, Die Damkbakterien des Sauglings, 1886. Esmarch, Centralb.
f. Bact., 1887. Ewart, Proceedings of the Roy. Soc., 1874. Falkenheim, Arch.
f. Exp. Patholog. u. Pharmakol., Bd. 19, 1885. Fasching, Centralb. Wiener
Akad. Wiss., 1891. Finkler and Prior, Deutsche Med. Woch. 1884. Fischel,
Zeitschr. f. Heilkunde, XII., 1891. Fischer, Zeitschr. f. Hygiene, 1887, 1888 ;
Centralb, f. Bacter., III]. and ]V. Fliigge, Microorganisms and Disease (New
Syd. Soc.). Foutin, Centralb. f. Bacteriolog., 1890. Forster, Centralbl. f.
Bacter., JI., 1887. Frankel and Franke, Knapp. Schweiger’s Archiv, XVII.
Frankland, P. and G., Zeitschr. f. Hygiene, 1889 ; Proc. Royal Soc., Vol. xlvii.,
1890, Frendenreich, Ann. de Micrographie, Vol. ii., 1890, and Vol. iii.,
1891. Frick, Virchow’s Archiv, CXVI. Friedreich, Virch. Arch., Bd. 30,
1864. Fulles, Zeitschr. f. Hygiene. Gaffky, Langenbeck’s Archiv f. Chir.,
BIBLIOGRAPHY, 663
Bd. XXVIII. Galtier, Compt. Rend., T. CVI., 1888. Gamaleia, Ann. de l'Institut
Pasteur, Vol. ii., 148%. Geddes and Ewart, Proceed. of the Roy. Soc. of London,
1878, Gessard, De la Pyocyanine et de son Microbe, 1882. Gessner, Archiv f.
Hygiene, Bd. 1X., 1889. Giard, Compt. Rend., 1882. Giltay and Aberson, Arch.
Neerland Sci., XXV., 1891. Globig, Zeitschr. f, Hygiene, IJ]. Gombert, Rech.
Exp. sur les Mic. des Conjunctives, 1889. Goodsir, Edinb. Med. and Surg,
Journ., 1842. Grotenfelt, Fortschr. des Med., Bd. VIT., 1889. Guillebeau, Ann.
de Micrographie, Vol. IV., 1892. Giinther, Deutsche Med. Woch., 1892; Arb.
a. d. Kaiserlich. Gesundh., 1893. Guttmann, Virch. Archiv, Bd. CVII. Hajek,
Berl. Klin. Woch., 1888. Hanser, Deutsch. Archiv. f. Klin. Med., Bd. XLII. ;
Ueber Faulnissbacterien, 1885. Heimer, Deut. Arch. f. Klin. Med., 1877.
Heinz, Centralbl. f. Bakt., V. Heller, Heller’s Arch. f. Chemie, 1847. Heri-
court and Richet, Compt. Rend., cvii., 1888. Heim, Arbeit aus. d. Ges., V.
Heydenreich, Centralb. fiir Bact., Bd. V., 1888. Holschewnikoff, Fortschr. der
Med., VII. Hueppe, Mittheil. aus d. Kaiserl. Gesund., IL, 1884. Itzigsohn,
Virchow’s Archiv, Bd. 13, 1858. Jacksch, Zeitsch. f. Phys. Chem., Bd. V.
Jaeger, Zeitschr. f. Hygiene, 1892. Johne, Deutsche Zeitschr. f. Thiermed. und
Path., Bd. X1J., 1886. Jordan, Rep, Mass. Board of Health, 1890. Jordan and
Richards, Report Mass. State Board of Health, Vol. ii., 1890. Karlinsky,
Centralb. f. Bacteriolog., Bd. V., 1889. Kartulis, Zeitschr. f. Hygiene, Bd. VI.,
1889, Katz, Centralbl. f. Bacter., ix., 1891. Kirchner, Zeitschr. f. Hygiene,
Bd. IX, 1891. Kitasato, Central. f. Bacteriolog., Bd. III, 1888. Kitt,
Deutsche Zeitschrift. f. Thiermed. und Path., Bd. XVIT., 1885. Klamann, All.
Med. Centralz., 1887. Klein, Centralb. f. Bakteriol. X. Koch, A. Habitita-
tims schr, Giéttingen, 1888; Mitth. aus dem K. Gesundheitsamte, Bd. IT. ;
Wundinfectionskrankheiten. 1878. Kolb, Arbeit. aus dem K. Gesundheits-
amte, Bd. VIII., 1891. Kramer, Oesterreich Landwirthschaft Centralbl.,
1891. Kriiger, Centr. f. Bacter., Bd. VII. Kurth, Botanische Zeitg., 1883 ;
Trans. Ninth Internat. Med. Congress, 1891. Kiischbert and Neisser, Bresl.
Artzl. Zeitschr,, 1883. Kiitzing, Linnea, 8, 1833. Lankester, Quar. Journ.
Mic. Sci., Vol. xiii, 1873, and xvi, 1876. Laser, Centralb. f. Bacteriolog.,
Bd. XL, 1892. Laurent, Ann. de l'Institut Pasteur. Lehmann and Neumann,
Bacteriolog, Diagnostik, 1896. Lépine and Roux, Compt. Rend., 1885.
Lesage, Bull. Mced., 1887. Letzerich, Zeitschr. f. Klin. Med., Bd. XIII. Leube
and Grasser, Virchow’s Archiv, Bd. C. Liborius, Zeitschr. f. Hygiene, I.,
1886, Lindenborn and Holschewnikoff, Fort. der Med. Lindner, Die Sar-
cineorg. der G&hrungsgeurche, 1888. Lingelsheim, Zeitschr. f. Hygiene.
List, Die Bakt. der Nutz- und Trinkwisser, 1888. Léffler, Berl. Klin. Woch.,
1887; Mitth. aus d. K. Gesund., Bd. II. Loeb, Centralb. f. Bacteriolog.,
Bd. X., 1891. Losdorfer, Med. Jahrb., Heft 3, 1871. Lucet, Ann. de VInstitut
Pasteur, Vol. v., 1891. Liiders, Ueber Abstammung u. Entwicklung des
Bacterium Termo, 1867. Lumnitzer, Centralb, f. Bakteriolog., Bd. III.
Lustgarten, Vierteljah. f. Derm. und Syph., 1887. Lustgarten and Manneberg,
Vierteljahresber. fiir Dermat. und Syph., 1887. Lustig, Diagnostik der Bakt.
d. Wassers. Macé, Traité pratique der Bacteriolog., 1892. Maierba, Giorn.
Intern. d. Sci. Med., 1888. Manfredi, Fortschr. der Med., 1886. Manneberg,
Centralb. f. Klin. Med., 1888. Marpmann, Ergiinzungshefte des Centr. f. Allgem.
Gesund., Bd. IJ. Maurea, Centralbl. f. Bakteriol., Bd. XL, 1892. Mendoza,
Centralbl. £. Bakteriol., Bd. VI. Menge, Centralbl. £. Bakteriol., Ba. VI, 1889 7
Bad. XIL., 1892. Miller, Deutsch. Med. Woch., 1884, 1886, 1887 : Microorganisms
of the Human Mouth, 1890, Miquel, Ann. de Micrographie, 1888-92. Mori,
Zeitschr. f, Hygiene, Bd. IV., 1888. Miilhduser, Virch. Arch., Bd. 97, 1884.
664 APPENDICES.
Munk, Virch. Arch., Bd. 22, 1861; Med. Centralbl, 1864. Muntz, Compt.
Rend., T. cxii. Neelsen, Beitrag. z. Biol. der Pflanzen, IIT. Neisser, Archiv f.
Hygiene, 1893. Neumann and Schaeffer, Virchow’s Archiv, Bd, CIX., 1887.
Nocard, Ann. de l'Institut Pasteur, 1887. Nocard and Mollereau, Ann. de
VInstitut Pasteur, T. 1, 1887. Oersted, Naturhist. Tidsskrift, Bd. III., 1840.
Okada, Centralbl. f. Bacteriolog., Bd. IX., 1891. Paltauf and Heider, Med.
Jabrbuch., 1889. Passet, Zeitschr. f. Hygiene, Bd. IX. ; Untersuch. u. die
eitr. Phleg. des Wiensch., 1885 ; Fortschr. der Med., 1888. Pasteur, Ann. de
Chim. et de Phys., T. 64, 1862; Compt. Rend., 1876; LII., 1861. Pansini,
Virch. Archiv, Bd. cxxii.. 1890, Perdrix, Ann. de l’Institut Pasteur, T. v., 1891.
Perty, Zur Kentniss. Kleinst. Lebensform, 1852. Pfeiffer, Deutsche Med.
Woch., 1888 ; U. die Bac. Pseudotuberculose bei Nagethiere, 1889 ; Zeitschr. f.
Hygiene, .Bd,.VI.; Bd. VIII, 1889. Plagge and Proskauer, Zeitsch. f.
Hygiene, IJ. Pohl, Centralb. f, Bacteriolog., 1892. Pommer, Mitth. des Bot.
Tost. zu Gratz., I. Popoff, Ann. de l'Institut Pasteur, 1890. Prazmowski,
Untersuch. ii. die Entwick. und Fermentwirkung einer Bakterien, 1880.
Prove, Beitr. z. Biolog. der Pflanzen, Bd. III. Raczynsky, Diss. der Militar.
Med. Akad., 1888. Reimann, Inaug. Diss. Wiirzburg, 1887. Rénon, Ann.
de l'Institut Pasteur, 1892. Roscoe and Lunt, Phil. Trans. Roy. Soc., 1892.
Rosenthal, Zeitschr. f. Hygiene, V. Roth, Zeitschr. f. Hygiene, Bd. VIII.,
1890. Russell, Zeitschr. f. Hygiene, Bd. XI., 1891. Sakharoff, Ann. de
VInstitut Pasteur, 1891. Sanarelli, Centralbl. Bd. IX. 1891. Scheibenzuber,
Allgem. Wien, Med. Zeit., 1889. Scheurlen, Deutsche Med. Woch., 1888.
Schimmelbusch, Deut. Med. Wocb., 1889. Scholl, Fortschr. der Med.,.VII.
Schottelius, Biol. Untersuch. ii. dem Micrococcus Prodig., 1887; Zeitschr.
f. Hygiene, 1890, Schréter, Beitriige z. Biol. d. Pflanz, Bd. 1, H. 2.
Schutz, Archiv f. Wiss. und Prakt. Thierheilk., 1888, Senger, Berl. Klin.
Woch., 1888. Smith, Central. f. Bakteriolog.,, Bd. X.; Med. News, 1887.
Sternberg, Report on Etiology and Prev. of Yellow.Fever, 1891; Manual
of Bacteriology, 1896. Strassmann and Strecker, Zeitschr. f. Meditinalbeamte,
1888. Suringar, Arch. Neerland, 1866 ; Bot. Zeit., 1866. Tappeiner, Fortschr ,
der Med., Bd. J. and II. Tataroff, Die Dorpater. Wasserbact., 1891. Tils,
Zeitsch. f. Hygiene, Bd. IX., 1890. Tizzoni and Giovannini, Zeigler’s Beitrige,
Bd. VI., 1889. Tommasoli, Giornale d. Mall. Ven. e. d. Pelle., 1889 ; Monat. f. Prakt.
Dermat., Bd. IX. Trambusti and Galeotti, Centralb. f. Bacteriolog., 1892.
Trécul, Compt. Rend., lxi. Tricomi, Atti. d. Soc, Ital. de Chir., 1887. Utpadel,
Archiv f. Hygiene, Bd. VI. Vandevelde, Studien zur Chemie des Bacillus Sub-
tilis; Zeitschr. f. Phys. Chemie, Bd. 8, 1884, Van Ermengem, Bull. Soc. Belge
de Microscop., 1888. Van Zieghem, Compt. Rend., Ixxxviii. and Ixxxix., 1879.
Vaughan, Amer. Journ. Med. Sci. 1892. Vignal, Archiv de Phys., viii., 1886 ;
Le Bacille Mesentericus Vulgatus, 1889; Archiv de .Phys., Vol. xxiii. Von
Besser, Beitr. z. Path, Anat., Bd. VI. Von Esmarch, Central. f. Bakteriolog.,
Bd. I. Warington, Journ. of Chemical Soc., 1890. Weibel, Central. f. Bak-
teriolog., 1887, 1888, 1893. Weichselbaum, Fortschr. der Med., Bd. V., 1887 ;
Ziegler’s Beitrige, Bd. IV., 1888; Das Osterreich. Samtiilswesen, 1889. Weisse,
Bull. Phys,-Mathemat, de St. Pétersbourg, III., 1845. Weisser, Zeitsch. f.
Hygiene, Bd. I., 1886. Welch and Nuttall, Bull. John Hopkins’ Hospital, 1892.
Welcker, Zeitschr. f. Ration. Med., 1859. Wimnogradsky, Ann. de l'Institut
Pasteur, 1890, 1891; Bot. Zeitung, 1893. Woodhead and Wood, Compt. Rend.,
T. cix., 1889. Zelinsky, Proc. Russ. Phys. Chem. Soc., 1893. Zopf, Sitz.-Ber.
-d, Botan. Ver. d. Prov. Brandenb.: Juni, 1882; Die Spaltpilzl. Zimmer-
mann, Die Bakt. und Nutz. u. Trinkwasser, 1890.
BIBLIOGRAPHY, 665
APPENDICES,
HEMATOZOA.
Bardels, Gaz. des Hépit., 1882. Crookshank, Journ. Roy. Micr. Soc.,
1886. Cuboni and Marchiafava, Arch. f. Exp. Pathol., Bd. 13, 1880; Atti della
R. Acad. dei Lincei., 1881. Danilewsky, La Parasitologie Comparée du Sang,
1889; Centr. f. Bakt. u. Parasitenk., 1891. Gerhardt, Zeitschr. f. Klin. Med.,
Ba. 7, 1884. Golgi, Archivio per le Scienze Mediche, 1886. Grassi and
Feletti, Centr. f. Bakt. u. Parasitenk., 1891. Klebs and Tommasi-Crudeli, Arch.
f. Exp. Pathol., Bd. 2,1879. Kotelmann, Virchow’s Arch., Bd. 97,1884, Laveran,
Compt. Rend., 1881; Ibid., No. 17, 1882; Traité des Fiévres Palustres, 1884.
Leoni, Gazetta Medica di Roma, 184, Mannaberg, Trans. New Syd. Soc.
1894, Manson, Brit. Med. Journ., 1896. Marchand, Virch. Archiv, Bd. 88,
1882. Marchiafava and Celli, Atti della R. Academia dei Lincei., 18&4 ;
Fortschr. d. Med., Bd. 3, 1885. Mariotti and Ciarrocchi, Lo Sperimentale, T. 54,
1884. Maurel, Ann. d’Hygiene, 1883. Richard, Compt. Rend., No. &, 1882.
Roszahegyi, Biol. Centralbl., Bd. 2, 1882. Sehlen, Fortschr. d. Med., Bd. II.,
1884. Sternberg, Buil. Nat. Board of Health: Washington, 1881. Tommasi-
Crudeli, Arch. f. Exp. Pathol., Bd. 12, 1880; Die Malaria von Rom., 1882 ;
Conférence faite 4 la 8, Sess. du Congrés Intern. Méd. & Copenhague, 1884.
Torelli, La Malaria in Italia, 1883. Ziehl, Deutsch. Med. Woch., Nr. 48, 1882.
COCCIDIA AND CANCER “ PARASITES.”
Albarran, Comp. Rend., 1889. Borrel, Archiv de Méd. Exper. et d’Anat.
Path., 1890. Bowlby, Brit. Med. Journ., 1891. Cazin, Internat. Congress of
Hygiene and Demog., 1891; Archiv Gen. de Méd., 1892. Darier, Compt.
Rend. Soc. de Biol., 1889. Delepine, Brit. Med. Journ., 1891 ; Trans. Internat.
Congress of Hygiene, 1891. Foa, Centralb. f. Bakt. u. Parasitenk., 1892.
Heidemann, Virchow’s Archiv., 1892. Leuckart, Parasites of Man,. 1886.
Loseh, Archiv f. Patholog. Anat., 1875, Nils Sjobring, Fortsch. der Med.,
1890. Noeggerath, Beitr. z. Struktur u. Entw. des Carcinoms., 1892. Pfeiffer,
Die Protozoen als Krankheitserreger, 1890. Podwyssozki and Sawtschenko,
Centr. f. Bakt. u. Parasitenk., 1892. Ruffer and Walker, Brit. Med. Journ.
1892; Journ. of Comp. Path., Vol. I. Russell, Brit. Med, Journ., 1890.’
Schiitz, Munch. Med, Woch., 1890. Shaltock and Ballance, Proc. Roy. Soc.,
1895. Soudakewitch, Ann. de l'Institut Pasteur, 1892. Wiekham, Archiv. de
Méd. Exper., 1890,
SUPPLEMENTARY APPENDIX.
EXTRACTS FROM THE FiNAL REPORT OF THE ROYAL
VACCINATION COMMISSION.
Tue final Report of the Royal Commission ito inquire into the
subject of Vaccination was published on September 18th, 1896.*
The author desires to gratefully acknowledge the permission granted
him, by the Controller of Her Majesty’s: Stationery Office, to make
extracts bearing more especially on the history and pathelogy of
protective inoculation and the prevention of small-pox. The reader
is recommended by the author to study the whole of the report.
History of Smatl-Pox.t
The early history of small-pox, like that of many similar diseases, is
obscure, is subject to much debate, and, save perhaps on one point, is of
antiquarian interest only.
The records of the eighteenth century show that the disease was very
prevalent in western Europe during the whole of that century. The
records of the seventeenth century also show that small-pox was a very
common disease during that century : this is especially the case as regards
the latter half of the century. The statistics which exist with respect to
Geneva, and various scattered statements, further show that small-pox
was a well-known disease in the sixteenth century ; but, except for the
records which are said to exist of severe epidemics in Iceland taking
place as early as 1241, as we go further back the evidence as to the
existence of the disease becomes less and less clear, and indeed debate-
able, depending as it does largely on the interpretation of incidental
*The report may be obtained either directly, or through any bookseller,
from Eyre & Spottiswoode, East Harding Street, Fleet Street, E.C., and 32,
Abingdon Street, Westminster, S.W.; or John Menzies & Co., 12, Hanover
Street, Edinburgh, and 90, West Nile Street, Glasgow; or Hodges, Figgis &
Co., Limited, 104, Grafton Street, Dublin.
+ The heading to the extracts from the Report of the Commission are mine.
—E. M. C.
667
668 SUPPLEMENTARY APPENDIX.
statements in various medical and other writings. There seems, how-
ever, to be adequate proof of the prevalence of small-pox in the East,
in Asia Minor and other countries, even in the earlier centuries of the
Christian era.
A view very generally taken teaches that small-pox, introduced from
the East, began to be common in western Europe during the fifteenth
century, though perhaps existing still earlier; that, it increased during
the sixteenth and seventeenth centuries, especially the latter ; and that it
was very prevalent during the eighteenth century.
In dealing with the eighteenth century it must be borne in mind that
during the second half of the century the natural conduct of small-pox,
as we shall see later on, was modified by the practice of inoculation—that
is, by the artificial giving of the disease by the introduction of the virus
through a wound in the skin.
Our knowledge of the history of small-pox in western Europe during
the seventeenth and eighteenth centuries is very largely based on the
official records known as the ‘London Bills of Mortality.” Official
records bearing on small-pox are furnished by Geneva, going back as
far as the sixteenth century, by Sweden, going back to the year 1749,
and by some other places. Data are also furnished, especially for the
latter part of the eighteenth century, by parish records in. various parts
of Great Britain reaching over a variable number of years, as well as by
scattered statements in various works.
These Bills of Mortality form by far the most complete source of our
knowledge of small-pox in England in past times ; but it must be borne
in mind that in respect to any contagious disease like small-pox the
conditions of London were peculiar. The population was to a marked
extent a moving one; a large number of persons were continually
entering London or leaving it, were passing to and from it, from and to
the provinces of England and other countries. Of these persons, some,
coming from infected districts, brought into London fresh sources of
contagion ; others again, coming from districts free from small-pox, and
never having had the disease, brought into London fresh material to serve
as food for the disease. Further, London presented in an exaggerated
degree the two features of a great city which have a great influence on
the progress and characters of a contagious disease like small-pox. The
crowding both of the dwelling-places and the thoroughfares, as well as
the movement continually going on, multiplied the opportunities for the
spread of disease, and the accompanying insanitary conditions, as well as
the greater inducement to irregular living, tended to increase the severity
of the disease when taken, and to heighten the mortality from it. The
history of small-pox in London must not be taken as representative of
the history of small-pox in England generally.
Inoculation of Small-pox.
The practice of inoculation for the small-pox—that is, the artificial
introduction of the virus into the system by the insertion of fluid from
a variolous pustule into wounds of the skin made for the purpose—
began definitely in England towards the end of the first quarter of the
REPORT OF THE ROYAL VACCINATION COMMISSION, 669
eighteenth century. Attention was directed to the matter by letters
from Timoni of Athens (dated 1713) and Pylarini, published in the
29th volume of the Philosophical Transactions (1716), and especially
by a letter from Lady Mary Wortley Montagu in 1717. Though
there are indications that in Great Britain and Ireland, as in other
countries, some sort of inoculation had occasionally been practised at
a much earlier date, the first clearly recorded case in England is that
of the daughter of Lady Mary Wortley Montagu (whose son had some
time before been inoculated at Constantinople), inoculated by Maitland,
in London, in April 1721. Other cases soon followed in England, and
about the same time the practice was also introduced in other countries
of western Europe, and into the United States of America, namely, at
Boston.
It was found that the attacks induced by inoculation were as a rule
milder, and very much less fatal, than the attacks of the “natural ”
disease, the fever and constitutional disturbance being less and of shorter
duration, and the eruptive pustules much fewer : the number of these
varied, being commonly a dozen or two, som:tines only two or three,
sometimes a hundred or more. In som: cases there was no eruption
at all, the effect being limited to constitutional disturbances and to
changes in the wounds of inoculation themselves ; it was maintained that
in such cases the disease had really been taken, and immunity against
a subsequent attack secured, as in case: of natural small-pox or of
inoculated small-pox manifesting itself in an eruption of pustules.
In England the practice of inoculation at its introduction, though
much lauded and strongly urged by some, was bitterly opposed by others.
Moreover, the initial enthusiasm in favour of it soon declined, so that
in the years 1730-40 very little inoculation seems to have been practised.
About 1740, however, a revival appears to have taken place: in 1746
an Inoculation and Small-pox Hospital was started in London ; and
during the whole of the latter half of the eighteenth century the practice
may be said to have been very general. It was especially so during the
last quarter of the century, the increase being at least largely due
to the ‘improved methods” of inoculation introduced by one Sutton
in 1763, and known as “ the Suttonian method.”
Since an inoculated person was infectious, each inoculation was
a source of danger to those, not protected by a previous attack, who
came into the company of, or even near, the inoculated person during
the attack; and this danger was increased by the fact that the mild
character of the inoculated disease permitted, in many cases at least,
the patient to move about among his fellows. Moreover, as Haygarth,
himself a zealous advocate of inoculation in a systematic regulated
manner, points out, the beneficial results of inoculation had robbed
the disease of its terrors to so great an extent that the rich and powerful
no longer made the efforts which they formerly did to prevent its.
entrance into, or its spread in, their neighbourhood, and thus Peon
its spread among the unprotected poor; so that inoculation though
eminently useful to the rich appeared to be injurious to the poor.
Adding, therefore, together the cases of inoculated small-pox, and the
670 SUPPLEMENTARY APPENDIX.
cases of natural small-pox of which the inoculated cases were in one way’
or other the cause, it seems probable that inoculation did tend to increase
the prevalence of small-pox ; but there are no recorded data to show that
this really was the case, and this supposed influence may have been
counterbalanced by other influences.
The evidence as to the influence which inoculation had on the
mortality from small-pox is in many respects conflicting. Haygarth,
though he admits that in other parts of the kingdom the practice may
have saved many lives, was persuaded that in his own part of England
and Wales the deaths by the small-pox had been augmented by it; and
he points out that in London, Geneva, and other “towns in different
situations and circumstances, the mortality from this distemper has
increased since the introduction of inoculation.” Several writers in the
_ latter part of the last, and the early part of the present century, held
a similar view. Other writers, again, opposed this view.
Tradition of the Dairy-folk.
There was at the close of the eighteenth century, if not earlier, in
districts where cow-pox had appeared, a belief among the dairy-folk that
those who had taken the cow-pox never took the small-pox ; and indeed
one Jesty, a Dorsetshire farmer, had in 1774, in the case of his wife and
sons, purposely introduced the matter of cow-pox into the human subject
with the view of protecting from small-pox.
Cow-pox.
Vaccinia or cow-pox is a disease affecting milch cows, and marked by
an eruption on the udder and teats. The disease can be communicated
from the cow to man. Dairymen and maids engaged in milking cows
affected with cow-pox are apt to have sores of a special kind on their hands
or elsewhere, the development of the sores being frequently accompanied
by febrile symptoms. There can be no doubt that, in a certain number
of cases at all events, such sores are the local manifestations of cow-pox ;
the virus from the eruption on the cow being introduced into some scratch
or other imperfection in the skin of the milker and there producing its
local effects, accompanied more or less by general symptoms.
Inoculation of Cow-poa.
The practice, however, of inoculating with the matter of cow-pox, or
vaccination as it was subsequently called, may be considered as dating
from the publication of the “Inquiry into the Causes and Effects of the
Variola Vaccine ” of Edward Jenner, published in the summer of the
year 1798. The practice rapidly spread, and prevailed widely in this
country and other parts of western Europe during the first quarter of the
present century. It was, beyond all question, so adopted in the genuine
belief that it afforded protection against small-pox.
In the treatise to which reference has been made Jenner records in the
first place a number (19) of cases in which a person who had accidentally
taken cow-pox from the cow had never had small-pox, and appeared
REPORT OF THE ROYAL VACCINATION COMMISSION, 671
incapable of taking that disease ; the insusceptibility being shown on the
one hand by the failure to contract the disease after ample exposure to
contagion, such as nursing and attending to or even sleeping with persons
suffering from small-pox, and on the other hand by the fact that when
the person in question was inoculated with the matter of small-pox in the
manner then usual (the matter being tested as to its efficiency on
susceptible persons) the inoculation failed to excite small-pox. In the
course of the inoculation practice it had been observed that when the
operation was performed upon a person who had already had small-pox,
either naturally or by inoculation, the wound of inoculation, instead of
developing, as it did when the operation was successful in a person who
had not had the small-pox, into a vesicle and so into a pustule with the
variolous characters (the development being accompanied by febrile
symptoms and, save in exceptional cases, by the appearance of a smaller
or greater number of variolous pustules on parts of the skin other than
the seat of inoculation), presented as a rule nothing more than some slight
inflammation, dying away in a few days without any other symptom, or even
healed at once without any symptoms at all, local or general ; and in the
exceptional cases in which further changes took place in the wound, these
were not accompanied or followed by an eruption of pustules or even by
the febrile and other general symptoms of small-pox. Accordingly, in
cases of small-pox inoculation where it was doubtful whether the disease
had been communicated, it had become not an uncommon practice to
repeat the operation, in order to judge by the effects produced whether
the earlier inoculation had or had not produced the disease ; and the
practice, thus originating in connexion with small-pox inoculation, had
come to be spoken of as the “ variolous test.”’
In his treatise Jenner distinguishes between what he calls true cow-pox
and other eruptions which he speaks of as spurious, and which he regarded
as not affording protection against small-pox, although he gives no details
to show that the cases quoted by him as affording protection were cases
of his true cow-pox. He also developed the view that matter derived from
horses suffering from the disease known as the grease is capable of giving
rise to cow-pox in the cow, and indeed is the real origin of the true
disease. It may be added that Jenner also expressed the opinion that the
protection thus afforded by cow-pox was permanent in character.
Jenner further recorded in the same treatise how he had in 1796
inoculated a healthy boy of eight years of age in the arm with cow-pox
matter taken from a sore on the hand of a dairymaid who had been
infected with the disease by milking cows suffering from cow-pox. He
describes the appearances subsequently presented by the wounds, and
states that, six weeks afterwards, the results of inoculating the boy with
variolous matter were those commonly seen to follow the inoculation of
persons who had previously had the cow-pox or the small-pox : that is
to say, the ‘‘ variolous test” showed the boy to be insusceptible to small-
pox, Some months afterwards the boy was again inoculated, but no
sensible effect was produced on the constitution. J enner then relates that
subsequently, in the spring of 1798, he inoculated a child, and obtained a
similar result with matter taken direvtly from the nipple of a cow infected
672 SUPPLEMENTARY APPENDIX.
with cow-pox ; from the pustule on the arm of this child he inoculated
another, and from this again several, and from one of these latter a fourth
in succession, and then a fifth. To three of these the “ variolous test”
was applied, and it is stated with the same results.
Woodville’s Lymph.
The experiences of Jenner did not stand alone. His results and
views attracted great attention, and in the early part of the year 1799
Woodville and Pearson, who were physicians to the Small-pox Hospital
in London, commenced making experiments with vaccine matter with
a view to ascertain whether it afforded protection against small-pox.
They arrived, like Jenner, at the conclusion that it did.
In January 1799 Woodville, having found cow-pox to be present in
a “dairy” at Gray’s Inn Lane, inoculated seven persons at the Small-
pox Hospital with matter from one of the cows at the “dairy,” and
other persons with matter from sores on a dairymaid employed at
the same place who had become infected from the cows, From these
cases he inoculated in succession others at the Hospital, eventually to
the number of many hundreds, and thus established the stock of what
has been spoken of as ‘ Woodville’s lymph.” Pearson also at the same
time occupied himself with the question of inoculation with the
cow-pox, writing a pamphlet about it. Woodville and he distributed
to many persons in this country and abroad quantities of the lymph
from the Hospital ; and this was the beginning of the more general
practice of vaccination, for Jenner’s stock of lymph, the results of
which he had described in his treatise, had come to an end.
Although Woodville’s ‘ Hospital lymph” appears to have been
widely distributed by himself and by Pearson, and thus to have been
the source of the lymph used in various places in the early days of
vaccination, it was not the only source, even in those days. Pearson
also obtained lymph from cow-pox at a dairy in the Marylebone Road,
and used this “in certain situations,” which may be presumed to
include places elsewhere than in the Hospital. He also speaks of
having obtained lymph from the cow from a third source. Jenner
again, who received and used some of Woodville’s Hospital lymph, also
obtained lymph from some other sources: for instance, from a cow
at a Mr. Clark’s farm in Kentish Town. Further, Woodville in 1800
speaks of his having at various times procured the vaccine virus as
‘produced in different cows, which when used at the Hospital produced
the same effects as the Gray’s Inn Lane lymph. We are not justified
in assuming that an account of every new source of lymph was
published ; and there may have been others, it is impossible to say
how many, than those just mentioned. In any case Woodville’s
Hospital lymph was not the only lymph used in those early days; not
only, however, was it largely used (indeed, we have no evidence of so
widespread a use of lymph derived from any other source), but the
use of it marks the definite beginning of the practice of. vaccination ;
and the history of it demands special notice.
REPORT OF THE ROYAL VACCINATION COMMISSION. 673
Of the cases recorded by Woodville in his Reports, the larger
number (about three-fifths)ipresented an important, and, as compared with
Jenner’s cases, a new feature, in that, in addition to the changes taking
place at the seat of inoculation and constituting what Woodville called
the ‘‘cow-pox tumour,’’ which may here be spoken of as the “ vaccine
vesicle,” an eruption over the body of a greater or less number of
pustules was observed. These eruptive pustules occurred in the very
first cases: of the seven cases inoculated from the cow, four, and of
the five inoculated from the dairymaid, four, had such pustules ; and
their appearance is recorded again and again in the series, down to the
case which appears last but one in the tabular statement forming part.
of the Reports. ’
Moreover, an eruption of pustules is described in certain of the cases.
of which accounts were published at about the same time by Pearson
and many others. In some of these cases the lymph used was supplied
from the Small-pox Hospital by Woodville or Pearson.
It must be admitted that these pustules were pustules of small-pox,
and that, therefore, Woodville’s cases, which did so much to establish
the practice of vaccination, were not cases simply of cow-pox but of
cow-pox mixed, so to speak, with small-pox. It has indeed been
maintained that Woodville’s cases were not cases of cow-pox at all—
that small-pox was inadvertently introduced into the very first cases ;
that the history of the whole series is the history of a series of small-pox
cases putting on special characters, and that therefore the lymph used
and distributed by Woodville and Pearson was in reality not cow-pox
lymph but small-pox. lymph. A review of all the evidence available
leads to no other conclusion than that, however much in Woodville’s,
Pearson’s and other cases cow-pox was mixed up with small-pox, the
lymph used and distributed by Woodville and Pearson and called by
them cow-pox lymph (excluding of course all the cases, of which there
were not a few, in which matter was taken not from the local “ cow-pox
tumour” at the seat of inoculation, but from one of the eruptive
pustules) was veritable cow-pox lymph having the true characters of
cow-pox lymph only.
It of course follows that the cases, both in Woodville’s practice and
jn that of others, in which the inoculation of cow-pox matter was.
accompanied by an eruption of pustules, due to small-pox being present
as well as cow-pox, when appealed to as showing immunity against
small-pox (by the test either of exposure to contagion or of inoculation),
furnished false evidence as to that immunity being due to cow-pox ;
it might have been due to the accompanying small-pox. So far then as
the adoption of vaccination was assisted by cases of this description, it.
may be held to have rested on erroneous data.
The Decline of Small-pox.
One effect of the introduction of vaccination was a very great decrease
in the practice of inoculation, which had become very prevalent during the
later part of the previous century. And the view has been ve forward
3)
674 SUPPLEMENTARY APPENDIX.
that, the prevalence of inoculation having greatly increased the amount
of small-pox, the diminution of small-pox in question was the result of
the decrease of inoculation.
The question how far the behaviour of small-pox in the eighteenth
century and earlier was influenced by sanitary conditions, is one rendered
difficult by the lack of exact information. We may distinguish between
overcrowding as one insanitary condition and all other insanitary condi-
tions, such as lack of cleanliness and the like. A priori we should ex-
pect that a dense population, especially one of great internal movement,
and one in continual interchange with surrounding populations, by
offering greater facilities for the conveyance of contagion, would lead to
a greater amount of small-pox. London was a conspicuous instance of
the above, and the apparent greater prevalence of small-pox in London
than in the provinces may be attributed to these causes: but it would
appear that the increase was felt—as indeed would, a priori, seem pro-
bable—rather in the constant presence of small-pox to a considerable
amount at all times than in the mortality of the epidemics when these
occurred. And the same seems also to be shown to a less extent in other
large cities, such as Liverpool. But in this matter of dense and moving
populations the eighteenth century did not differ markedly from the
early part of the nineteenth, We might @ priori expect the other
acknowledged imperfect sanitary conditions of the eighteenth century to
increase the fatality of, and so to a corresponding extent the mortality
from, small-pox ; but there is no exact evidence to confirm this supposi-
tion. If on the contrary we recognise that in the course of the eighteenth
century the general mortality, the relative number of deaths from all
causes, went on decreasing, and attribute, as has been done, this decrease
to improved sanitary conditions, no like decrease of small-pox took place.
Again, the places which were deemed the most salubrious appear to have
been visited by epidemics of small-pox as severe as those which fell on
unhealthy places. Thus the epidemic in Chester in 1774 was undoubtedly
a severe one, and yet Haygarth writes, ‘‘The healthiness of Chester,” as
shown by statistics, must appear so very extraordinary as to be almost
incredible.” And in general both the incidence of, and mortality from,
small-pox seem to have been far less affected by sanitary conditions than
might a priori have been expected.
It may be urged against the view that the decline of small-pox was
due to improved sanitary conditions, in the first place, that, admitting
the introduction of sanitary improvements, no evidence is forthcoming
to show that during the first quarter of the nineteenth century these
improvements differentiated that quarter from the last quarter, or half,
of the preceding century in any way at all comparable to the extent
of the differentiation in respect to small-pox. In the second place,
admitting & priori that crowded dwellings tend to increase the liability to
contagion, and so the prevalence of the disease, while other insanitary
conditions tend in addition to increase the fatality among those attacked,
so that insanitary conditions as a whole must tend to increase the
mortality from small-pox,—no evidence is forthcoming which distinctly
shows that the dependence of the prevalence of, or the mortality from,
REPORT OF THE ROYAL VACCINATION COMMISSION. 675
small-pox, on the lack of sanitary conditions, was a feature of the history
of small-pox during the eighteenth century.
Upon the whole, then, we think that the marked decline of small-pox
mortality in the first quarter of the present century affords substantial
evidence in favour of the protective influence of vaccination,
Age Incidence of Small-pox.
A study of the age incidence of small-pox mortality is very instructive.
In connexion with this point it is necessary to bear in mind that experi-
ence has led to the conclusion that, whatever be the protective effect of
vaccination, it is not absolutely permanent ; the most convinced advocates
of the practice admit that after the lapse of nine or ten years from the
date of the operation its protective effect against an attack of small-pox
‘rapidly diminishes, and that it is only during this period that its power in
that respect is very great ; though it is maintained that, so far as regards
its power to modify the character of the disease and render it less fatal,
its effect remains in full force for a longer period, and never altogether
ceases. The experience upon which this view is founded is derived almost
exclusively from the case of infantile vaccination. It has been supposed
by some that the transitory character of the protection results from
-changes connected with the growth from infancy to adult years. Whether
this be so or not, we have no means of determining.
No doubt, when Jenner drew the attention of the public to the value
-of vaccination, he believed that a single successful inoculation of vaccine
matter secured absolute immunity for the future from an attack of small-
pox. It is certain that in this he was mistaken. It may well be doubted
whether the anticipation was a reasonable one. No such immunity is
secured by an attack of small-pox, though there are few who would
maintain the proposition that it is without protective influence against
another attack. A priori there would seem to be no sound ground for
expecting that vaccinia would afford more potent protection than small-
pox itself. The extent of the protection afforded (assuming that there is
‘some protective influence) could only be determined by experience. It soon
became apparent that Jenner had, in the first instance, overrated the
effect of vaccination. That he should thus have overestimated it
is not to be wondered at, when the tendency to be unduly sanguine,
which besets the discoverer of any new prophylactic, and, indeed, every
discoverer, is borne in mind. ;
We think, taking it all together, that the evidence bearing upon
the question whether the vaccinated are less liable to be attacked by
small-pox than the unvaccinated, points to two conclusions: first, that
‘there is, taking all ages together, less liability to attack among the
vaccinated than among the unvaccinated ; and next, that the advantage in
this respect enjoyed by vaccinated children under ten years of age is
greatly in excess of that enjoyed at a more advanced period of life.
In considering whether vaccination has been the principal cause of the
decline, we must inquire whether the other causes suggested by those who
deny the efficacy of vaccination will satisfactorily account for it.
676 SUPPLEMENTARY APPENDIX.
Effect of Sanitation.
It is said that the decline has, in the main, been due to changes in the
general conditions of life in the different parts of the United Kingdom,
apart from the spread of the practice of vaccination,—amongst other
things, to improvement of sanitary conditions.
It is beyond doubt that an infectious disease like small-pox is, other
things being equal, more likely to spread in towns than in country districts,
and more likely to spread in crowded town districts than in others not so
densely populated ; so that we should expect a lessened proportion of
overcrowded dwellings, by diminishing the opportunities for contagion,
to check the prevalence of the disease and consequently to render its
mortality less.
Effect of Isolation.
It has been maintained that the decline in small-pox mortality is
largely due to more frequent and systematic attempts to isolate those
suffering frora small-pox. We think an answer to this contention is to be
found in the fact that it is only in quite recent years that there has been
any systematic practice of isolating small-pox patients, and that it has
been confined even then to a very limited number of localities. The
fact to which we are about to call attention in greater detail than
hitherto, that the decline in the deaths from small-pox is found almost.
exclusively among those of tender years, appears also to militate against.
the contention. The risk of contagion is not confined to children.
Adults also are subject to it. If a better system of isolation had been
a main cause of the reduced mortality, we should have expected to see it.
operate in the case of adults as well as of children. . At the same time
we are far from thinking, as will appear when we come to deal with
that subject, that the efforts at isolation which have characterised recent.
years have been without a beneficial effect on small-pox mortality.
Sanitary Legislation.
We have already pointed out that on & priori grounds it is reasonable
to think that improved sanitary conditions would tend to diminish
the fatality of, and so to a corresponding extent the mortality from,
small-pox. And there can be no doubt that the period with which
we are dealing has been characterised by an improvement of this
description. There has been better drainage, a supply of purer water,
and in other respects more wholesome conditions have prevailed.
Tt may be useful at this point to furnish a brief summary of the
principal Sanitary Acts which have been passed relating to the different.
parts of the United Kingdom.
In 1848 was passed the first great and comprehensive measure which
may be called the groundwork of our sanitary legislation as regards
England. The Public Health Act of 1848 was, however, principally
designed for towns and populous places in England and Wales, not
including the Metropolis, which was dealt with in Acts passed in the
same year. The powers of local government supplied by the Act were
generally an extension of those before given by sundry local Acts to
REPORT OF THE ROYAL VACCINATION COMMISSION. 677
Commissioners of Sewers in the Metropolis, and to authorities in a few
large towns. Many provisions corresponding to sections in the Towns
Improvement Clauses Act of 1847 are found in the Public Health Act,
and communities were thus enabled to obtain by a simple process powers
which they could not previously obtain except by a local Act incorporating
sections of the Towns Improvement Clauses Act.
In 1848 was also passed the Nuisances Removal and Diseases Preven-
tion Act of that year, in substitution for a similar Act of 1846 which was
about to expire; and in 1849 this Act of 1848 was amended. The
provisions of all these three Acts extended to England, Scotland and
Ireland. In 1855 a comprehensive Nuisance Removal Act was, as regards
England, substituted for the Acts passed in 1848 and 1849; and in the
following year there was similar legislation for Scotland. In 1860 the
English Act was amended ; and in 1866, by the Sanitary Act of that year
(to which we shall again refer), the provisions of the English Acts of 1855
and 1860, as then amended, were applied to Ireland.
In 1855, by the Metropolitan Local Management Act of that year,
provision was made for the appointment of a medical officer of health
and an inspector of nuisances by every vestry and district board in
the Metropolis. This provision did not extend to the City of London,
where, in 1848, a medical officer of health had been appointed under
power given by a local Act.
In 1858, the Local Government Act of that year, to be construed with
the Public Health Act of 1848 as one Act, was passed, and took effect in
all places where that Act was in force at the time of its passing ; and,
as regards England, these two Acts together constituted until 1872 the
principal sanitary legislation on the statute-book.
There followed, however, within the next ten years many public Acts
having sanitary objects, some applying to all, and some to particular, parts
of the United Kingdom, besides numerous other Acts of local application.
We need only now specially refer to one of these public statutes—the
Sanitary Act of 1866, which was probably the most important, and applied,
in part at least, to England, Scotland and Ireland, This Act, amongst —
other things, extended the powers of local authorities for the disposal of
sewage, and, in amending the English Nuisances Removal Acts of 1855
and 1860, added to the definitions of nuisances, especially as regards
crowded houses and workshops, and to the duties and powers of local
authorities for their abatement, especially in the way of providing means
for disinfection and places for the reception of dead bodies.
In 1867 the Public Health (Scotland) Act was passed—a comprehensive
measure which consolidated into one Act, with certain amendments, the
whole statute law relating to the public health in Scotland.
In 1872 a complete distribution of England into sanitary districts took
place, and some further amendments were made in the sanitary laws. In
1875 these laws were consolidated in the Act of that year. In 1891
a Sanitary Act was passed relating to the Metropolis.
In 1874 an Act was passed for Ireland, containing substantially the
game provisions as those which had been enacted in the case of England
in 1872.
678
SUPPLEMENTARY APPENDIX.
Value of Vaccination.
We have not disregarded the arguments adduced for the purpose of
showing that a belief in vaccination is unsupported by a just view of the
facts. We have endeavoured to give full weight tothem. Having done so,
it has appeared to us impossible to resist the conclusion that vaccination.
has a protective effect in relation to small-pox.
We think :—
1.
2%
3.
That it diminishes the liability to be attacked by the disease.
That it modifies the character of the disease, and renders it
(a) less fatal, and (0) of a milder or less severe type.
That the protection it affords against attacks of the disease is.
greatest during the years immediately succeeding the operation
of vaccination. Jt is impossible to fix with precision the
length of this period of highest protection. Though not in
all cases the same, if a period is to be fixed, it might, we think,
fairly be said to cover in general a period of nine or ten years.
. That after the lapse of the period of highest protective potency,
the efficacy of vaccination to protect against attack rapidly
diminishes, but that it is still considerable in the next quin-
quennium, and possibly never altogether ceases.
. That its power to modify the character of the disease is also
greatest in the period in which its power to protect from
attacks is greatest; but that its power thus to modify the
disease does not diminish as rapidly as its protective influence
against attacks, and its efficacy during the later periods of life
to modify the disease is still very considerable.
. That re-vaccination restores the protection which lapse of time
has diminished ; but the evidence shows that this protection
again diminishes, and that, to ensure the highest degree of
protection which vaccination can give, the operation should be
at intervals repeated.
. That the beneficial effects of vaccination are most experienced
by those in whose case it has been most thorough. We think
it may fairly be concluded that where the vaccine matter is.
inserted in three or four places, it is more effectual than
when introduced into one or two places only—and that if the
vaccination marks are of an area of half a square inch, they
indicate a better state of protection than if their area be at all
considerably below this. .
Question of Specific Protection or of Antagonism.
When an attack of disease secures immunity or protection against.
another attack of disease, the two attacks are, as a rule, attacks of the
same disease. Some pathologists have, it is true, of late years been led to
suppose that one disease may confer some degree of immunity or protec-
tion against another different disease ; but instances of this are few, and,
moreover, cannot be regarded as thoroughly established. The ordinary
instances of immunity are so clearly those in which the attack, natural or
REPORT OF THE ROYAL VACCINATION COMMISSION. 679
artificial, which confers the immunity is of the same disease as that towards
which immunity is conferred, that identity of disease has been considered
as eon to the conferring of immunity. And it has been argued that
it is a priori improbable that cow-pox should confer immunity from small-
pox, seeing that the two are different diseases. Such a purely theoretical
argument can have little weight against positive evidence of vaccination
having actually conferred immunity. If this be definitely proved to be the
fact, proof is thereby at the same time afforded that the theory is unsound,
either because a particular disease may confer immunity against a different
disease, or because small-pox and cow-pox are not different diseases. For
the practical object with which alone we are concerned, it is not material
that we should reach any conclusion upon the question what is the real
source of error in the theory alluded to, supposing it to be erroneous ?
We shall content ourselves, therefore, with a very brief notice of the
subject.
It appears to us that we may dismiss for practical purposes the
theoretical questions which were discussed before us so fully. If the fact
be established that the introduction of vaccine matter and the consequent.
vaccinia produce some effect upon the human body which renders it less
susceptible to small-pox, or which modifies that disease when the small-pox
virus enters the system, it will not be a strange or unwonted experience
that we should be unable to explain how this comes about. Science
has not yet succeeded in freeing therapeutics or kindred subjects from
obscurity, or in solving all the problems which they present. The precise
modus operandi by which a previous attack of a disease furnishes security
against a subsequent attack by the same disease has not yet been elucidated.
There can be no cause for astonishment, then, if we are unable to trace
the steps by which vaccination exerts a protective influence, supposing
the fact that it does so be established, nor is it essential that we should
succeed in tracing them. Our inability to accomplish this does not seem
to us to be the slightest reason for regarding with doubt the conclusions
to which the facts lead us. :
Professor Crookshank, than whom no one has more strongly insisted on
the theoretical arguments against the protective influence of vaccination in
relation to small-pox, gives it as his opinion that vaccination creates a
transient antagonism to that disease. We understand his view to be that
an attack of disease can only afford protection against the same disease,
and that small-pox and cow-pox are not the same but different diseases.
We gather, however, that, in his opinion, so long as the state of antagonism
lasts, the person in whose system it exists is less likely to suffer from
small-pox than he would be if the state of antagonism were wanting.
This seems to us to amount in effect to the same thing as saying that.
during that period vaccination has conferred some protection. Whether
the effect be to create antagonism or to confer protection, and whatever
difference there be between the modus operandi in the one case and in the
other, we know equally little about it. If a condition of transient
antagonism to small-pox is induced by vaccination, theoretical considera-
tions will not afford a guide of the slightest value to the conclusions how
long this transient antagonism will last, or how soon it will pass away.
680 SUPPLEMENTARY APPENDIX.
Experience, and experience alone, can answer that question. A priori we
do not see that there is any better reason for supposing that it would last
for two or three years than that its duration would extend to ten years.
Cow-pox and Small-pox not Convertible.
Jenner himself, in his first paper, advanced the view that the cow-pox
and small-pox were identical with each other; and since his time
numerous observers have attempted to prove the identity of the two
diseases experimentally—namely, by giving rise to cow-pox in the cow
through the inoculation of small-pox matter, or by the introduction of
contagion in other ways. It may at once be stated that while cow-pox
is readily transferred from the cow to man and back again from man to
the cow, the disease in man being identical with that in the cow, small-
pox cannot be transferred from man to the cow so as to give rise toa
disease in the latter identical in its features with the small-pox of man.
Nor can cow-pox be so transferred to man as to give rise in him to
small-pox. The two diseases are not in this sense convertible.
Small-pox Vaccine.
In most cases the attempt to transfer small-pox from man to the
cow has had simply a negative result ; no obvious effect of any kind has
been observed. This has been the case in the attempts to introduce the
contagion through absorption by the respiratory or digestive organs, and
in most of the attempts to introduce the contagion by inoculation. In
certain instances these latter attempts have produced results which may
be briefly described in three categories. (We may pass over the isolated
experience of Thiele, who in 1838 asserted that by keeping small-pox
matter sealed between glass plates for ten days before using it, and
by diluting it with milk when using it for inoculation, the matter thus
treated through ten removes through the human body—the cow not
intervening at-all—was converted into something which gave results
identical with those of ordinary vaccine matter. We are not aware of
any attempt to corroborate this experiment.)
The first category includes the experiments in which the inoculation
of small-pox matter into the udder, or adjoining parts, of the cow gave
rise at or near the seat of inoculation to a vesicle, either identical in
visible characters with the ordinary vaccine vesicle produced by inocula-
tion with the matter of cow-pox, or to a vesicle the features of which,
while not corresponding wholly with those of a perfect vaccine vesicle,
so closely resembled them as to justify the vesicle being called a vaccine
vesicle. Further, the matter from a vesicle which at the first inoculation
had not the characters of a perfect vaccine vesicle, when carried through
a second or third remove in the cow, fully acquired those characters, and
when transferred to man gave results indistinguishable from the ordinary
vaccine vesicle. Indeed, lymph of such an origin has come into general
use for vaccination purposes. Of the experiments, the best known or
most quoted are those of Thiele (1838), Ceely (1840), Badcock (between
1840 and 1860), Voigt (1881), Haccius and Eternod (1890), King (1891),
REPORT OF THE ROYAL VACCINATION COMMISSION. 681
Simpson (1892), and Hime (1892); but there are several others. The
details of the experiment are very scanty in the cases of Thiele and
Badcock, but more full in the others, especially, perhaps, those of Ceely
and Haccius.
In the second category may be placed the experiments of Klein and
Copeman. Klein, who had in 1879 obtained in 31 trials what then
appeared simply negative results, found in a renewed research in 1892
that the result of the first inoculation in the cow of small-pox matter
was not a distinct vesicle but merely a thickening and redness of the
wound. Lymph pressed from the thickened wound produced, when
inoculated into a second cow, a like result, but rather more marked ;
the thickening and reddening still further increased with a third and
a fourth cow. Lymph squeezed from the wounds of the fourth cow
produced in a child typical vaccine, and crusts from the child produced
typical vaccine in a cow. Copeman obtained somewhat similar results ;
the appearances increasing in three removes and approaching those of
typical vaccine, but not reaching them.
The third category consists of the results obtained in an elaborate
inquiry conducted by a Commission of the Society of Medical Sciences at
Lyons, with Chauveau at its head. Those results, reported in 1865, were
briefly as follows :—
Inoculation of the cow with small-pox matter in any one of the 30
animals used did not give rise to a vaccine vesicle. Nevertheless a
definite result was obtained ; in the form, however, not of a vesicle, but
of a thickening and inflammation of the wound ; when a puncture was
employed this became a papule. Lymph squeezed from such a papule
and inserted into a second animal gave rise to a like papule ; and this,
again, might be used for a third animal, but often failed; and the effect
could in no case be carried on through more than three or four removes.
When the inoculation was repeated on an animal in which a previous
inoculation had produced such a papule, no distinct papule was formed ;
and, moreover, lymph squeezed from the seat of inoculation produced no
effect at all when used for the subsequent inoculation of another animal.
This shows that the development of the papule was the result of the
specific action of the virus: The same is shown by the fact that no such
papule was produced when the small-pox matter was inserted into an
animal which had previously had cow-pox naturally or artificially, as well
as by the fact that when an attempt was made to vaccinate, with vaccine
matter of proved efficacy, an animal on which a papule had been so
developed by inoculation with small-pox, the vaccination failed, though
the animal had never had natural cow-pox or had never been vaccinated.
The specific nature of the lymph of the papule is further shown by the
fact that such lymph when used'on the human subject gave rise to verl-
table small-pox. It has been urged that in this case the virus producing
the effect was simply the old virus used in the inoculation, producing
the papule and still clinging to the wound. This is disproved by the
experience that lymph from a papule of the second remove also gave rise in
the human subject to veritable small-pox.
Thus Chauveau and his Commission found that small-pox implanted
682 SUPPLEMENTARY APPENDIX.
in the cow gave rise to a specific effect which was not cow-pox btt was.
of the nature of small-pox, though its manifestations in the cow were
different from those of small-pox in man. They also obtained similar
results in attempting to transfer small-pox to the horse.
It must be admitted that the results finally obtained in some of the
successful cases were indistinguishable from those of vaccination ; the:
characters of the local vesicle, the absence of eruptive pustules and of
contagiousness, show that the lymph thus apparently originating from
small-pox in the hands of Ceely, Badcock, and others, was vaccine lymph.
It has been urged that a vaccine vesicle making its appearance in the
wound of inoculation with small-pox was due to the accidental intro-
duction of cow-pox matter into the wound; the small-pox matter in
the wound produced no effect, and the cow-pox matter its usual effect.
Several considerations support this view. The cow is peculiarly suscep-
tible to cow-pox. In some cases (Ceely, Voigt), the animal was
vaccinated as well as inoculated with small-pox: thus, in Ceely’s first
case, the animal was inoculated with small-pox on one side of the body,
and a few days after vaccinated on the other side. In many cases the
experiments were conducted in an animal vaccine establishment, the
stalls, the operating tables, and the assistants being those used or engaged
in vaccination. It is true that in some cases at least special precautions,
sterilisation of instruments and the like, were taken to avoid the accidental
introduction of cow-pox ; but in observations of this kind the difficulties
of avoiding all such sources of error are notorious. Still the successful
cases are now so numerous that it is difficult to resist the conclusion that
the same accident could not have occurred in all, and that a transformation
of small-pox into cow-pox—that is to say, into the artificially inoculated
cow-pox which we call vaccine *—really took place.
Accepting this view provisionally, it may be remarked that in most.
cases the transformation was sudden and complete ; the small-pox virus,
under the influence of the tissues of the cow, became immediately
converted into vaccine virus, and this produced a typical vaccine vesicle.
In some cases (ea. gv., that of Hime) the transformed virus produced its
effect not in the wound of inoculation, or not chiefly so, but at some little
distance from it. In some cases the characters of the vesicle first formed,
though sufficiently distinct to justify the vesicle being called a vaccine
vesicle, were not those of a perfect vaccine vesicle, but the lymph from
such a vesicle, at least after one or two removes, gave rise to most typical
vaccine vesicles.
In Klein’s experiments the transformation was gradual. In his fourth
cow, the virus was as yet not typical vaccine, since it did not produce a
typical vesicle ; yet it was so far already vaccine that, transferred to the
child, it produced typical vaccine (unless we suppose some accidental
introduction of vaccine to have taken place). That the vesicle on the
child was vaccine, and not small-pox unaccompanied by eruptive pustules,
was shown not only by its characters but also by the fact that lymph from
it produced typical vaccine in the cow.
In Chauveau’s experiments no transformation at all took place.
* The italics are mine.—E.M.C.
REPORT OF THE ROYAL VACCINATION COMMISSION. 683
As has been urged in another place, there are no adequate reasons
leading us to believe that in the human subject the small-pox virus and
the cow-pox virus can so act on each other as to form a hybrid disease.
But this does not preclude the view that, accepting the conclusion that
the body of the cow has the power to convert small-pox into vaccine, the
virus may exist for a while in a phase in which, while ceasing to be
typical small-pox, it has not yet fully acquired the characters of vaccine,
and we may regard Klein’s results as illustrating this. In some of the
experiments—for instance, those of Ceely and Voigt—the results obtained
with the lymph of the vesicle produced by the inoculation of small-pox
give rise to the suspicion that the lymph had small-pox qualities, as seen,
for example, in the case of Ceely’s assistant, Taylor ; but the facts cannot
be said to be more than suspicious—they are not decisive. Moreover,
admitting that the vesicle itself in such cases was the result of the trans-
formed virus, some not transformed old virus might still remain dormant
in the wound, and might be present in the lymph of the vesicle, mixed
with the transformed and generating virus; this old virus might have
happened to be in excess on the point of the lancet which wounded
Taylor.
Smaill-pox Vaccine—Cow-pox Vaceine—Horse-pox Vaccine—Cattle-
plague Vaceine—Sheep-pox Vaceine.
Taking all the various facts into consideration, we seem led to the
provisional conclusion that under certain conditions the tissues of the
cow are able to transform small-pox into vaccine, that these conditions
may be such as to lead to the transformation being sudden and complete,
that under certain other conditions the transformation may be gradual
and incomplete, and that under certain other conditions (and these seem
most commonly to obtain) the transformation into vaccine does not take
place at all. But what the above conditions are has not as yet been
- clearly made out. It has been suggested that one condition favourable
to the transformation is extreme youth of the subject: to effect the
change the animal used should be a calf of not more than three or four
months old ; but this is not definitely proved.
Until these favourable conditions have been clearly recognised, so that,
the conditions being fulfilled, the transformation will always be secured,
the conclusion cannot be regarded as indisputable. Moreover, it must
be borne in mind that effects more or less closely resembling a vaccine
vesicle have been at various times obtained by various observers through
inoculating man or the cow or another animal with material other
than that obtained from the pustules of the small-pox of man. Much
discussion has taken place concerning the “ grease ” of the horse, which
Jenner believed to be the origin of the cow-pox of the cow. Without
entering into any discussion of the matter, it may be said that investiga-
tion has shown that horses do suffer from a malady which, transferred to
the cow, gives rise to a vaccine identical apparently with that produced
by the inoculation of the natural cow-pox. Hence this malady is spoken
of as the “horse-pox,” and some cases at least of so-called ‘ grease”
appear to be cases of this horse-pox. But it is at least not proved that
684 SUPPLEMENTARY APPENDIX.
all the cases of “grease” which by inoculation were found to give rise to
vaccine vesicles in man were cases of true horse-pox. And this at least
must be said, that no investigations as complete and varied as those which
have been carried out with regard to the development of vaccine vesicles
through the inoculation of small-pox matter, have been carried out with
regard to the alleged development of vaccine vesicles by the inoculation
-of other material, such as the matter from the eruptions of the sheep-pox,
the cattle plague, and the like. Nor have there been like extended in-
-quiries as to the production of vesicles resembling those of vaccine by the
inoculation of small-pox matter into animals other than the cow or the
horse ; such results as have been obtained by observers are conflicting.
‘There is still room for much inquiry ; meanwhile it may be said that, in
any case, the evidence in favour of a possible transformation of small-pox
into vaccine is.sufficiently strong. to remove the force of the theoretical
objection to the power of vaccination to secure immunity towards small-
pox, on the ground that the two diseases are absolutely distinct.
Risks of Vaccination.
It must not be forgotten that the introduction into the system of even
a mild virus, however carefully performed, is necessarily attended by the
production of local inflammation and of febrile illness. If these results
did not in some measure follow, the practice would probably fail in its
protective influence. As a rule, the inflammation and illness are of a
trifling character,; in exceptional cases, however, they may exhibit more
severity, and, as certain facts submitted to us in evidence have shown,
there are cases, though these are rare, where a general eruption may
follow vaccination.
In order to determine how far the risk of erysipelas is a necessary
incident of vaccination, what is the extent of that risk, and how it may
best be avoided, it is necessary to consider the various circumstances
which may occasion erysipelas and allied diseases in the case of vaccinated
children. It is established that lymph contains organisms, and may
contain those which under certain circumstances would be productive of
erysipelas. It is therefore possible that some contagious material (the
specific virus of erysipelas, for instance,) may be conveyed at the time of
vaccination, owing either to its presence in the lymph employed, or to its
being conveyed by the vaccinator himself, or by those with whom the
‘child comes in contact at the time of vaccination. We believe that the
cases in which the virus is conveyed at the time of vaccination are rare.
It has, however, in some instances been clearly established, the immediate
occurrence of erysipelas in several co-vaccinees making it practically
certain that some virus was conveyed at the time of the operation. In
some instances where this has been the case, and there is every reason for
believing that the contagion was conveyed through the medium of the
lymph, it is nevertheless in evidence that the vaccinifer did not display
anything more than a slightly inflamed arm. The scrupulous avoidance
of inflamed arms in vaccinifers will do much to reduce the risk of
conveying erysipelas in the act of vaccination (a risk which, as we have
REPORT OF THE ROYAL VACCINATION COMMISSION. 685-
Seen, has been proved to be a very slight one), but it is possible it would
not wholly remove it.
We have dwelt upon features presented by the cases of erysipelas and
various forms of septic disease which have followed vaccination, because-
they suggest precautions which may be adopted to lessen, if not to.
prevent, such evils in the future. If, for example, vaccination were
performed at the patient’s home instead of ata public vaccination place,.
the chance of disease being contracted at the time of vaccination would
be to some extent diminished ; and the same may be said of the inspection
of the vaccinated person which takes place eight days after the operation.
On these points we shall have some remarks and recommendations to-
make at a later stage of our report.
A study of the cases which have been made the subject of careful
examination and report points to the conclusion that an exercise of
greater care would largely diminish the risk, already small, of erysipelas-.
contagion and blood-poisoning.
Although it may be confidently hoped that by additional care on the
part both of vaccinators and parents, the number of inflamed arms and
of cases of erysipelas may be reduced to very few, yet it is not to be
expected that such occurrences will be wholly prevented. A vaccination
wound is, like one from any other cause, so long as it exists, a source of
some risk.
The use of calf-lymph, though it may be supposed to be more free-
from the risk of conveying erysipelas, does not appear to prevent inflamed
arms. Some witnesses have indeed supposed that it is attended with
more risk of inflammation than the employment of that taken from the.
human subject. This opinion has not, however, been corroborated by
some of those of widest experience.
Nothing has produced so deep an impression hostile to vaccination as
the apprehension that syphilis may be communicated by it. It was at
one time doubted whether syphilis could result, and it was even confi-.
dently asserted that it could not. The fact that this was possible had
been fully established, and was generally acknowledged by the medical
profession, before we commenced our inquiries.
The very close resemblance in certain very rare cases of the results
of vaccination, whether with calf-lymph or humanised lymph, to those
attributed to syphilis (a resemblance so close that it has caused in a few
cases a difference of opinion whether the disease was syphilis or vaccinia)
has led to the expression by Dr. Creighton of the opinion that there is
some essential relationship between the two diseases. This, however,.
is a point of speculative, almost it might be said of transcendental
pathology, upon which for practical purposes it is useless to enter. Tt
must be sufficient to remark that, whatever may be the relationship.
alluded to, it exists, if it exists at all, equally between small-pox and
syphilis as between vaccination and syphilis. For all practical purposes.
variola and vaccinia are both wholly distinct from syphilis, and their
differences are, with the rarest exceptions, easily recognised, They are
alike in being attended by affections of the skin and mucous membranes,
and exceptionally by disease of the bones, eyes, and other parts ; but in all.
686 SUPPLEMENTARY APPENDIX.
these it is a question of resemblance and not of identity with which we
have to deal.
Only a few items of the evidence produced before us appear to require
special notice : among these, the most prominent is what has been known
as the “Leeds case,” . upon which we have heard the evidence of Mr.
Ward, Mr. Littlewood and Dr. Barrs. The witnesses named regarded it as
a case of syphilis, conveyed by vaccination, but all of them admitted that
the course of events was most unusual. We have carefully investigated
this case, and notwithstanding the opinion formed by the witnesses, there
appears good reason to doubt whether it was one of syphilis. The case was
made the subject of careful inquiry by Dr. Barlow on our behalf, who
shared the doubt we have expressed. The view taken by the medical
inspector of the Local Government Board who in the first instance in-
vestigated the case was that it was a case of hereditary syphilis. It seems
certain, however, that the parents of the child whose death was in question
were not in any way affected with syphilis. The vaccinifer also appeared
to be free from any taint of that disease, and its family history confirmed
this view. The co-vaccinees from the same lymph also exhibited no trace
of syphilis. These facts of themselves make out a strong case against
that having been the nature of the disease. Coupled with the fact that
it could not have been communicated by the vaccinator himself, they seem
to render it practically impossible that syphilis was the cause of death.
If the symptoms exhibited had in all respects corresponded with those
which are known to characterise syphilis, the proper inference might have
been that there was some error in ascertaining the facts of the case. But
it is beyond question that the course of events was very different in some
respects from that experienced in undoubted cases of syphilis, and we
think the true conclusion is that it was nota case of that disease. It may
probably be classed with a few others as examples of gangrene and blood-
poisoning, the direct result of vaccination, which are not to be explained
by supposing the introduction of any syphilitic or other poison. Fortu-
nately, such cases are extremely rare—so much so that the witnesses
concerned knew of no case precisely parallel.
The evidence offered to us would lead to the belief that, whilst with
ordinary care the risk of communication of syphilis in the practice of arm-
to-arm vaccination can for the most part be avoided, no degree of caution
can confer an absolute security. The rejection as vaccinifers of young
‘infants, say below four months of age (in whom congenital syphilis may
be as yet undeclared), and of adults (in whom the disease may possibly
have been recently acquired) are precautions which would probably shut
out almost the whole of the risk. The outbreaks of syphilis in connection
with vaccination which have been mentioned to the Commission (all of
which had been previously published) have occurred chiefly in.arm-to-arm
vaccination amongst soldiers, or from the use as vaccinifers of young
infants the offspring of parents whose history was not known to the
vaccinator. It must, however, be admitted that neither the examination
of the vaccinifer if taken alone, and without a knowledge also of the
parents, nor the most scrupulous avoidance of any visible admixture of
blood with the lymph, are in themselves, however valuable, sufficient
\
REPORT OF THE ROYAL VACCINATION COMMISSION. 687
absolutely to exclude risk. The evidence given by Dr. Husband, of the
Vaccine Institution of Edinburgh, established the fact that all lymph,
however pellucid, does really contain blood cells. Absolute freedom from
risk of syphilis can be had only when calf-lymph is used ; though where
the antecedents of the vaccinifer are fully ascertained, and due care is
used, the risk may for practical purposes be regarded as absent.
It is obvious that the employment of ealf-lymph only would wholly
exclude the risks as regards both syphilis and leprosy. Respecting the
latter disease, however, there appears to be reason to doubt whether any
risk exists, and at any rate it does not concern the British population.
Even in leprosy districts the employment of English human lymph would
be, so far as leprosy is concerned, as safe as that from the calf.
There can be no doubt that vaccination ought to be postponed when
-erysipelas, scarlet fever, measles, or chicken-pox are prevalent in the
neighbourhood of the child’s residence, or, if the child is not to be
vaccinated at home, either there or near the place of vaccination. Here
again there would be « gain if the home was more often the place of
vaccination.
It would, in our opinion, be an advantage if the postponement of
vaccination were expressly permitted, not only on account of the state of
the child, but of its surroundings and any other conditions rendering the
operation at the time undesirable. If more discretion in this respect
were possessed and exercised, we think untoward results would become
even rarer than they are.
We are quite alive to the objections which may be urged against a
prolongation of the period within which vaccination must be performed.
Jt will naturally be said that a number of children, who otherwise would
‘be protected against small-pox, would be left without that protection,
and would thus be liable to suffer from the disease themselves, and be a
source of danger to others. It must be remembered, however, that so
long as children cannot walk, the risk of their contracting contagion is
less than if they were able to move freely about and mix with other
people, and that, for the same reason, the risk of their communicating
-contagion to others is less, We cannot trace in the statistics relating to
Scotland any grounds for believing that the later compulsory vaccination
cage which prevails in that country.as compared with England has affected,
to any substantial extent, the general small-pox mortality of Scotland,
though no doubt it may have led to some deaths among children under
six months of age which otherwise would not have taken place.
We have already shown how satisfactory a position Germany has
occupied in relation to small-pox since the year 1874. The age of com-
pulsion in that country is the end of the next calendar year after birth.
It is true that re-vaccination has been there made compulsory as well as
primary vaccination ; but we think the experience of Germany is not
without its bearing on the question we are now considering. Wherever
the line is drawn, whether at three months or six months, it will always
leave a class of unvaccinated persons. The age to be fixed isa question
of policy into which many considerations must enter. If an extension
of the age within which vaccination was required rendered its untoward
688 SUPPLEMENTARY APPENDIX.
incidents fewer in number, and diminished hostility to the operation, it-
may be that on the whole it would promote the cause of vaccination, and
secure, as its result, that the number of vaccinated persons would be
greater than at present.
Means, other than Vaccination, for diminishing the Prevalence of Small-pox ;
and how far such means could be relied on in place of Vaccination.
Another question upon which we are asked to report is, what means,
other than vaccination, can be used for diminishing the prevalence of
small-pox ; and how far such means could be relied on in place of
vaccination.
The means, other than the inoculation of small-pox or cow-pox, which
have been referred to by witnesses as being capable of diminishing the
prevalence of small-pox, are such means as have been employed against
infectious diseases generally ; they may be summarised as—(1) Measures
directed against infection, ey., prompt notification, isolation of the
infected, disinfection, etc. ; (2) Measures calculated to promote the public
health, the prevention of overcrowding in dwellings or on areas,
cleanliness, the removal of definite insanitary conditions, etc.
The principle underlying the practice of isolation with its accompany-
ing machinery is obviously the very opposite of that which recommended
the practice of inoculation ; it aims at exclusion of the disease, whereas.
inoculation aimed at universal acceptance by artificially “sowing or
“buying” the disease. Except in regard to the plague, our knowledge
and practice of measures of isolation and quarantine against epidemics is
of relatively recent growth. As the result of increased knowledge of the
mode of propagation of infectious diseases, of greater sanitary activity,
and under the stimulus of legislation, organised effort, more or less
thorough, is now, in this as in other countries, directed against the spread
of dangerous infectious diseases. Side by side with a vaccination system,
means of isolation, etc., have been successfully employed to check the
‘spread of small-pox. They have also been sometimes so employed in
recent years in places where vaccination has fallen into disuse.
It will be well to commence with a brief statement of the growth of
our knowledge on the subject of isolation as a means of dealing with
infectious or contagious diseases. We have already adverted to the fact-
that small-pox is highly contagious, and that contagion from those
suffering from it is the means by which the disease is propagated,
Although reference to infection appears in some of the Arabian writers,
the contagiousness of small-pox attracted little attention in this country
and in western Europe until the eighteenth century. Sydenham (1624-89),
though he refers to the contagiousness of small-pox, did not dwell upon
the matter, and did not regard it as so important an element in the spread.
of the disease as some peculiar constitution of the atmosphere to which
he attributed epidemics. Boerhaave was the first, at the commencement
of the eighteenth century, distinctly to formulate the now generally
accepted doctrine that small-pox arises only from contagion.
In 1720, Mead drew up an elaborate system of notification, isolation,
disinfection, etc., in view of a threatened invasion of the plague ; but no
REPORT OF THE ROYAL VACCINATION COMMISSION. 689
attempt to deal with small-pox in a similar fashion appears to have been
made until the last quarter of the eighteenth century. This was in all
probability largely due to the adoption of inoculation as the recognised
defence against small-pox, and the acceptance of Sydenham’s doctrine
of epidemic causation may have exercised an influence in the same
direction.
No writer appears to have suggested methods of isolation, disinfection,
ete., against small-pox prior to 1763. In that year Rast of Lyons published
his “ Reflections on Inoculation and Small-pox, and upon the means which
might be taken to deliver Europe from that malady.” He maintained—
(1) That small-pox was not a necessary and inevitable malady ; (2) That it
arose only from contagion ; (3) That it resembled plague in most of its
features. His conclusion was expressed in these terms: ‘“‘T say, that to
“deliver Europe from small-pox we must act upon principles directly
“ opposed to inoculation ; far from multiplying the contagion, we must
“keep it away by taking the same precautions and employing the same
“ measures against that malady as have proved so successful against leprosy
“and the plague.”
The earliest account of the practical employment of such means is
from Rhode Island, U.S.A. Haygarth, on the authority of Drs. Moffat and
Waterhouse, states that for many years prior to 1778 small-pox had been
successfully prevented from becoming epidemic there by regulations for
isolation of the infected on a neighbouring small island specially used for
that purpose, and for quarantining infected vessels, destruction of infected
clothing, etc. Moreover, inoculation was discouraged at Rhode Island,
and those who wished to be inoculated had to go to some place away from
the Island, and were not to return until there was no danger of their
infecting others.
A passage in Dimsdale’s work on Inoculation, published in 1781, shows
that in some towns of England pest-houses were beginning to be used for
small-pox. In 1784 Haygarth, of Chester, published his “ Inquiry how to
prevent the Small-pox,” and in 1793 “A Sketch of a Plan to exterminate
the Small-pox from Great Britain.”
The great epidemic of smiall-pox at Chester in 1774, to which allusion
has already been made, was the occasion of Haygarth’s first attempts at
organised dealing with epidemics of small-pox with a view to preven-
tion. In his “Inquiry” he combated Sydenham’s doctrine that epidemics
are due to some occult condition of the atmosphere, and argued that
small-pox was always spread by infection only. He further maintained
that the variolous poison could be carried as an infection for a little
distance only through the air, and “ consequently that the small-pox may be
“ prevented by keeping persons liable to the distemper from approaching
“within the infectious distance of the variolous poison till it can be
“destroyed.” These views led him, upon the return of an epidemic in
1777, to propose a plan for the prevention of the natural small-pox, and
in 1778 a society was formed to carry out the plan in Chester. The plan
consisted on the one hand of a general inoculation at people’s homes at
stated intervals, on the ground that the inoculated small-pox was far less
fatal or injurious than the natural small-pox, and on the other hand of
44
690 SUPPLEMENTARY APPENDIX.
“Rules of Prevention” based on Haygarth’s views of infection. In th
report of the Society, called shortly ‘‘The Small-pox Society,” date
September 1782, it is stated that in the four and a half years of it
existence two general inoculations had been held, and that the death
from small-pox had been greatly lessened. Great difficulties, howevei
were met with. ‘“ A large proportion of the inhabitants” refused inocula
tion, and a large proportion also, ‘‘ being fearless, or rather desirous, tha
their children should be infected with the natural small-pox,” refused t
obey the Rules of Prevention. Hence, though the same report state
that the example of Chester had been followed by Liverpool, wher
“a general inoculation was successfully executed in the autumn of 178
and another in the spring of 1782,” and in Leeds, where a genera
inoculation was held in 1781 and another proposed in 1782, with sucl
success that the Royal College of Physicians in Edinburgh appointed :
committee to inquire into “ the modes of conducting the general inocula
tions of the poor” thus adopted in these places, the plan met with sucl
difficulties that it was ultimately abandoned. It will be observed that ;
general inoculation was an essential part of the plan proposed anc
carried out in 1778-82; but, writing in 178+, Haygarth looked forwarc
to being able ultimately to dispense with inoculation, and in the prefac
+o his later edition, published in 1793, he states more definitely that th:
adoption of his Rules of Prevention without any general .inoculatior
might exterminate small-pox in some country other than Great Britain
It must be remembered, however, that Haygarth entertained the opinior
that the infection of small-pox could not be carried through the air above
about half a yard, and that no one could be infected by the clothes of :
person visiting a small-pox patient provided that he kept beyond thi:
distance from the patient. It is obvious that if this had been establishec
the control of the disease by isolation would be a much simpler matte:
than it really is.
In the Medico-Chirurgical Review for 1796 there appeared an accoun’
of a work by Dr. Faust, of Leipsic, entitled “An Essay on the Duty ol
“Man to separate persons infected with the Small-pox from those ir
‘Health, thereby to effect the extirpation of that disease equally fron
“the towns and countries of Europe,” in which it was argued that the
first person ill in a place is the only source from which all the rest
perhaps hundreds and thousands, become affected, and that if he wer«
put immediately into a situation where he could not injure by contact
those who had not had the disorder, the spread of the disease would be
‘prevented.
In the same Review for 1799 appeared an account of establishments
for the extirpation of small-pox. The failure of inoculation to attain the
‘desired end is referred to, and legislation is urged to facilitate isolation
lt is further stated that in 1796 the Prussian College of Physicians re
ported favourably to the King on the project, and that at Halberstadt ii
had been resolved to establish a house for the purpose. At Cédte d’Or ix
France a similar plan had been tried with success.
In 1798 Jenner’s “Inquiry” was published, and in the early years of thi:
century inoculation began to be discouraged ; for a while the prospects o1
REPORT OF THE ROYAL VACCINATION COMMISSION. 691
annihilating small-pox by vaccination appear to have superseded, in the
minds of many, the plans of Haygarth and others. Some vaccinators,
however, like Willan and Ring, still looked to methods of quarantine and
to national and municipal regulations promoting isolation to exterminate
the small-pox.
It is worthy of notice, too, that Haygarth himself, in a letter quoted
by Dr. Cappe of York in a communication to the London Medical and
Physical Journal (vol. iv., p. 429), dated October 13th, 1800, remarked,
“ An introduction of the vaccine still more than of the variolous inocula-
tion would effectually promote the great object of my publications.”
Prior to the year 1866 there was no provision made by law for enabling
sanitary authorities to establish hospitals for infectious diseases, and thus
to promote the isolation of such cases. The only institutions of that
description then existing were the result of private effort. So far as
regards small-pox there was, practically speaking, no provision for its
treatment by means of isolation.
The Sanitary Act of 1866 empowered, though it did not compel,
local authorities throughout England and Wales, Scotland and Ireland,
to provide or to join in providing isolation hospitals for the use of the
inhabitants of their districts. There was further legislation on the
subject by the Public Health Act, 1875; the Public Health (London) Act,
1891 ; the Public Health (Scotland) Act, 1867 ; and the Public Health
(Ireland) Act, 1878, into the details of which it is not necessary to enter.
The most recent Act relating to the matter is the Isolation Hospitals Act
of 1893, which applies to the small towns and rural districts of England
and Wales. .
Stamping-out System in Leicester.
Leicester suffered severely from small-pox in 1872, 346 deaths having
been registered as caused by it. Two deaths from that disease occurred
in 1873, but no other until 1877, when there were six, and one in the
following year. The next year in which small-pox deaths were registered
was 1881. There were two in that year, and five and three in the follow-
ing years. No other death took place until 1892 and 1893, in which
years the fatal cases numbered 21. ;
Prior to 1875 the vaccination laws were well observed in Leicester.
In that year the number of children born who were unaccounted for was
only some 4 per cent. Since then there has been, as we have seen, a
marked and progressive decline in the number of vaccinations, especially
since 1883, until at the present time 80 per cent. of the children born
remain unvaccinated.
The borough hospital for infectious diseases was erected in 1871-2
outside the town; though within the last few years houses have been
built in proximity to it. It appears to have been with Dr. Crane, the
Medical Officer of Health in 1875, that the quarantining the inmates of
an infected house, in addition to isolating the patient, originated. His
successor, Dr. Johnston, established it in 1877 as a regular system. He
was aided in this, after 1879, by the notification of infectious diseases
then rendered compulsory by a private Act which Leicester, anticipating
692 SUPPLEMENTARY APPENDIX.
most other towns, obtained in that year. Dr. Johnston reported that up
to 1884 the spread of small-pox from imported cases had been arrested in
20 instances by the means he adopted.
His successor, Dr. Tomkins, though, like his predecessors, regretting
the increasing disuse of vaccination, bore testimony in his annual reports
to the efficacy of the measures adopted in Leicester, and expressed his
opinion that had such a system been in force at Sheffield in 1887 it would
not have suffered in the way it did.
In 1892 small-pox became prevalent in different parts of England,
especially in Lancashire and Yorkshire. Many of the large provincial
towns suffered, and Leicester amongst them. There were, in 1892-3, 357
cases of small-pox in Leicester, of whom 21, or 5°8, died ; 193 households
were invaded, containing 1234 persons. The first importation was by a
tramp, whose disease, passing unrecognised, caused infection at a common
lodging-house and at the workhouse. Eleven other importations of the
disease by tramps occurred in the course of 1892-3.
Leicester suffered less than many of the other large towns which have
been invaded by small-pox during recent years, both in the number of
cases and in the number of deaths. In connection with this, however,
a point to which we have already called attention must be borne in mind.
The disease was remarkably slight there in its fatality, even as regards
those who, by reason of their age, could not be affected by the change of
practice in relation to vaccination. Dr. Priestley, the Medical Officer of
Health, claims, in his report to the Sanitary Committee for 1893, that
it was by reason of the energetic methods adopted that the disease had
been prevented running riot through the town. His claim may be well
founded. At all events, the experience of Leicester affords cogent
evidence that the vigilant and prompt application of isolation, etc., even
with the defects which were brought to light during the recent epidemic,
is a most powerful agent in limiting the spread of small-pox. It is true
that the system and appliances which appeared adequate for some years
failed to prevent a serious outbreak of small-pox in 1892-3. We think
its value was none the less real.
Stamping-out System in London.
In the Report of the Royal Commission of 1881, already alluded to,
suggestions were made with regard to notification and isolation which
have since been largely carried into. effect. As we have said, it was con-
sidered proved that the existing small-pox hospitals had caused a spread
of the disease in their neighbourhood, We cannot but think that this
may in some measure account for the greatly increased mortality from
small-pox in London during the 1871-72 epidemic as compared with the
rest of the country. It is true that the statistics relating to England and
Wales outside the Metropolis include those of other large towns where the
same evil was present ; but it probably did not exist there in so aggravated
a form, and the effect may be neutralised by the statistics relating to
smaller towns and rural districts with which they are combined. This
idea has been suggested to us, as the result of the inquiry, how it has come
about that whilst the Metropolis, in the decennium 1867-76, and again
REPORT OF THE ROYAL VACCINATION COMMISSION. 693
down to 1885, compared so unfavourably with the rest of the country, the
condition has since that date become so entirely changed? We think it
is impossible to attribute this change to vaccination. There is no reason
to suppose that the position of the Metropolis in respect to vaccination
has, since the year 1885, become superior to the rest of England and
Wales: rather the other way, as the decrease in infantile vaccination has
been greater during the last few years than in the rest of England and
Wales. The change, therefore, must be due to some other cause.
The hospitals which, in the opinion of the Commissioners, were
propagating the disease in their neighbourhood, were in operation
down to July 1882, when their Report was made. In 1877 and 1878, and
again in 1881, small-pox was epidemic in London to a considerable extent.
We have stated in detail in paragraph 471 * the steps which were taken
by the Metropolitan Asylums Board in consequence of the recommenda-
tions of the Royal Commission. It will be seen that the intra-urban
hospitals still continued in use, and that complaints were made in 1884 that
they were spreading small-pox in their vicinity, although the number in
each of them was not allowed to exceed 50. In October 1884 this number
was reduced to 25. It was not, however, until 1885 that the system now
in operation was inaugurated, and all cases of small-pox were treated in
hospital ships. . It is impossible not to be struck with the fact that it is
since the year 1885 that the Metropolis has presented so satisfactory an
aspect as regards small-pox mortality. The facts to which we have been
calling attention certainly seem to point to the conclusion that this has
been due to a system of isolation, well organised and administered, the
beneficial effect of which is no longer neutralised by a spread of the
disease from the hospitals in which the isolation is carried out.
Upon the whole, we think the experience of London affords cogent
evidence. of the value of a sound system of isolation in checking the
spread of small-pox. ;
Stamping-out System in Australia.
The experience of isolation systems in Australia is interesting and
worthy of special notice, because whilst in this country the quarantining
of persons who have come in immediate contact with those suffering from
small-pox has only been possible with the consent of the persons whom it
was proposed to subject to quarantine, in Australia their removal to a
place of isolation has been made compulsory.
Australia, by virtue of its geographical position, and the consequent
separation by long sea voyage from infected ports, enjoyed for a long
time a sort of natural isolation. Thus, Hirsch, in his “ Historical and
Geographical Pathology,” vol. i., pp. 133-4 (1881), remarks :—
“The continent of Australia up to 1838 had enjoyed an absolute
“jmraunity from small-pox ; towards the end of that year the disease
“appeared at Sydney, having been imported probably from China ; it
“Jasted, however, only a short time, and remained absent from the
« continent until 1868. In that year it was introduced into Melbourne by
« a ship, and again it spread only to a slight extent, and quickly died out.
* Final Report.
694 SUPPLEMENTARY APPENDIX.
“By a rigorous inspection of ships on their arrival, it has been found
“possible to prevent subsequent importations, a notable instance of pre-
“ vention having occurred in 1872. Tasmania has hitherto quite escaped
“the disease ; so also has New Zealand, where an importation of it in
“(1872 was prevented by strictly isolating a vessel that had arrived with
* small-pox on board.”
In New South Wales, Dr. MacLaurin, who has been President of the
Board of Health since 1889, informed us that the Government act on the
assumption that small-pox is an exotic disease, and that every case must
have come from outside the colony, and it is therefore dealt with under
a quarantine Act of William IV., originally instituted for dealing with
cholera. By an Act passed in 1882, notification of small-pox was made
compulsory on medical men and householders under heavy penalties. At
Sydney notification of small-pox is followed up by the compulsory
removal of the patient and all persons who have been in the house with
the patient to the quarantine station at North Head. This station is 670
acres in extent, and situated on the peninsula at the mouth of Sydney
Harbour, and is seven miles from the Health Office, with which there is
telephonic and telegraphic communication. The persons are conveyed to
the station by a steamboat comfortably fitted expressly for the purpose,
and no difficulty has been experienced in effecting their removal. It was,
in Dr. MacLaurin’s opinion, by carrying out this practice of isolation and
quarantine that ‘‘ the epidemic of 1881-82 was suppressed,” and small-pox
‘‘has never become epidemic since this plan has been adopted.” The
persons who have been in the house with the patient are detained 21 days
in quarantine from the date of the last possible contagion. Should a case
of small-pox arise among them, those who had been in contact with such
infected person would be detained for a further period of 21 days, and so
on. To facilitate this, the exposed persons are distributed in separate
groups within the station. They are allowed to receive letters or parcels,
etc., and a telegraph operator is employed, ‘‘ whose special business it is
“to work the telegraph at their request.” ‘“ Reasonable compensation is
“given by the Government for loss; ” and there are heavy penalties under
the original Act whereby the quarantine is secured. The station is, accord-
ing to Dr. MacLaurin, “a pleasant place to stay in, and everything is done
“that can be done to make the people comfortable : they have nothing
“whatever to do, and are free from all care, and they can spend the day
“pleasantly enough ; but they do not like it.” No one, however, raises
any objection to the Sydney system : “the people are all very sensible
about it.” In all Australian towns the same system is carried out as
strictly, with the result that there was not a case of small-pox in Australia
on February 5th, 1890 ; and Dr. MacLaurin is of opinion that the risk of
dying of small-pox in Australia is smaller than in any other part of the
world. As regards vaccination :—In New South Wales it is very little
practised ; there is no compulsory Act ; and though medical opinion is in
favour of it, an opinion shared by Dr. MacLaurin, it is not likely that a
compulsory Vaccination Act could be passed or would be tolerated. The
proportion of young persons in New South Wales who are not vaccinated
is accordingly very large ; probably much more than half of those under
REPORT OF THE ROYAL VACCINATION COMMISSION. 695
ten years of age are unvaccinated. Although Dr. MacLaurin favours
vaccination and respects it highly, he is satisfied that the system of isola-
tion as supervised by him is perfectly successful. As President of the
Board of Health he considered it his business to produce extinction of the
disease ; he does not consider vaccination a sufficiently absolute protection
for such purpose ; and he is “ fully of opinion that the only way in which
“you can bring to an end an outbreak of small-pox, that is to say, bring
“it under control, and not leave it to work itself out, is by notification
‘‘and isolation. Of course, in any small community, if you let the disease
‘in it will work itself out in time, because all the susceptible people will
“have had it; but the only way in which you can absolutely control an
“epidemic of small-pox is by a system of notification and isolation.”
Small-pox has never been epidemic in Western Australia. Only one
case has occurred within the last 31 years, and that was an imported one;
quarantine was carried out, and no infection occurred ; the immunity from
the disease is mainly at least due to isolation. Before 1879 vaccination
was not generally practised—a great majority of those born in the colony
were unvaccinated ; in that year a compulsory Vaccination Act was passed
in consequence of Sir H. Ord and Dr. Waylen’s representations, and in
consequence of reports of small-pox in other colonies, and not on account
of the existence of small-pox in Western Australia.
In Tasmania there was a compulsory vaccination Jaw, but it was found
to be inoperative because no one was appointed to conduct the prosecu-
tions, and it has now fallen into desuetude. The same system of isolation
and quarantine is exercised as in the other Australian colonies; Small-pox
was for the first time introduced into Tasmania in 1887, and although
preparations for isolation were inadequate, the disease was soon stamped
out. Communication between Launceston and Melbourne was temporarily
suspended, and to this precaution the non-invasion of Victoria was
attributed. The particulars of this, the first introduction of small-pox
into Tasmania during the history of that colony, are to be found in a
report to the Central Board of Health by Mr. A. Mault, dated Novem-
ber 17th, 1887. The origin of the outbreak is not clear, but it was
presumed to have been imported, probably by a ship from China, into
Launceston. The earliest case reported to the Local Board of Health
was on September 23rd, though it appears that earlier cases had passed
unnoticed, or had been notified as measles. Thirty-three cases in all
occurred, every one of which was traced to direct infection from the first.
case. By September 27th a temporary hospital had been erected, and
thither patients and suspects to the number of 72 were removed. The last
case appeared on October 13th. Other persons who had been to the infected
houses were isolated in ‘their houses and watched. Only four of the 47
persons quarantined at the station were attacked. The clothing was
burnt, and very thorough disinfection of the infected houses was carried
out, and the dead were interred in a special cemetery. The other colonies
were communicated with, and quarantine, at first unduly rigid and after-
wards relaxed, was practised against ships proceeding from Tasmania.
Although vaccination had been nominally compulsory in Tasmania, it was
estimated that two-fifths of the population were unvaccinated.
696 SUPPLEMENTARY APPENDIX.
Suggested Stumping-out System in the United Kingdom.
We have no difficulty in answering the question, what means other
than vaccination can be used for diminishing the prevalence of small-pox ?
We think that a complete system of notification of the disease, accom-
panied by an immediate hospital isolation of the persons attacked, together
with a careful supervision, or, if possible, isolation for sixteen days of
those who had been in immediate contact with them, could not but be
of very high value in diminishing the prevalence of small-pox. It would
be necessary, however, to bear constantly in mind, as two conditions of
success : first, that no considerable number of small-pox patients should
ever be kept together in a hospital situate in a populous neighbourhood ;
and secondly, that the ambulance arrangement should be organised with
scrupulous care. If these conditions were not fulfilled, the effect might
be to neutralise or even do more than counteract the benefits otherwise.
flowing from a scheme of isolation.
When we turn to the other branch of the inquiry, haw far such means
could be relied on in the place of vaccination, we find ourselves involved
in questions of a much more complicated matin: We have little or no
experience to fall back upon. The experiment has never been tried.
The nearest approach to a trial of it has probably been in Australia. But
even in the parts of that country to which we have alluded the population
has not been entirely unvaccinated, though there has been a large
unvaccinated class amongst it. Moreover, in applying the experience of
Australia to this country, two things must be borne in mind. In the first
place small-pox has only appeared from time to time, introduced from
without at one or other of the ports of the country, and the several
colonies of which Australia is composed are of great territorial extent,
with few large centres of population. In this country small-pox is
always present in some part of it. There has not been a single year
without several deaths from the disease. Large centres of population
are numerous, and the intercourse between them constant. In the several
colonies of Australia the number of ports is not great, the vessels which
enter them are comparatively speaking not numerous, and the ports from
which they arrive are many days’ voyage distant ; and there are careful
arrangements for quarantining vessels to exclude disease. The shipping
which enters English ports is of vast quantity, and passengers are brought
in large numbers from the continent of Europe not only daily, but it may
almost be said hourly ; the voyage, too, is but brief. The other matter to
be remembered is, that part of the Australian system is the compulsory
removal to quarantine for 21 days of thése who have been in the house
with the patient, in addition to the transfer of the patient himself toa
hospital. There can be no doubt that such a system, if completely carried
out, would be of the highest efficacy. But it is obvious that in this
country the practical difficulties of working such a scheme in the large
towns would be really insuperable, to say nothing of the difficulty of
procuring legislative sanction for it.
REPORT OF THE ROYAL VACCINATION COMMISSION. 697
Value of Isolation.
We can see nothing, then, to warrant the conclusion that in this country
vaccination might safely be abandoned, and replaced by a system of
isolation. If such a change were made in our method of dealing with
small-pox, and that which had been substituted for vaccination proved
ineffectual to prevent the spread of the disease (it is not suggested that it
could diminish its severity in those attacked), it is impossible to contem-
plate the consequences without dismay.
To avoid misunderstanding, it may be well to repeat that we are very
far from underrating the value of a system of isolation. We havealready
dwelt upon its importance. But what it can accomplish as an auxiliary
to vaccination is one thing ; whether it can be relied on in its stead is quite
another thing.
Even admitting fully the protective effect of vaccination, it does not,
in our opinion, diminish the importance of measures of isolation or dis-
pense with their necessity. We think that steps should be taken to secure
a more general provision for the isolation of small-pox patients than
exists at present. We have already called attention to the fact that
mischievous results are likely to follow the use as a small-pox hospital of
a building situate in 4 populous place. We think that wherever it is
placed it should have sufficient space around it to enable the sanitary
authority to add rapidly to the accommodation by the erection of tem-
porary buildings.
Compulsory Provision of Isolation Hospitals.
Sanitary authorities are now sometimes reluctant to provide isolation
hospitals. We think that, on a petition by a prescribed number of the
ratepayers in a sanitary district, the Local Government Board, if satisfied
that the hospital accommodation ought to be provided, should have power
to make an Order for such provision.
Compulsory Notification of Small-pox.
We think that notification of small-pox should everywhere be compul-
sory, and, whenever the disease showed a tendency to become epidemic,
a notice should be served by the sanitary authority upon all persons in
the neighbourhood who would be likely to come within the reach of con-
tagion, urging them to submit to vaccination or re-vaccination, as the case
might be, if they had not been recently successfully vaccinated or re-
vaccinated ; and attention should be called to the facilities afforded for
their doing so. Attention should also be called to the importance of
avoiding contact with persons suffering from the disease, or coming into
proximity to them, and of avoiding contact with any person or thing
which may have become infected. It is important to notice that, even
where vaccination has been neglected, there is great readiness to submit
to it in the presence of a threatened epidemic ; a large number of vacci-
nations are then obtained willingly and without opposition. Whenever
698 SUPPLEMENTARY APPENDIX.
a sanitary authority has received notification of a case of small-pox,
we think the fact should be at once communicated to the vaccination
authority of the district in which the case of the disease has occurred.
Regulations as to Tramps, Inmates of Lodging-houses, ete.
Our attention has been drawn to the circumstance that outbreaks of
small-pox have not unfrequently had their origin in the introduction
of the disease to common lodging-houses by tramps wandering from place
to place. In view of this we make the following recommendations :—
(i.) That common shelters which are not now subject to the law
relating to common lodging-houses should be made subject.
to such law.
(ii.) That there should be ‘power to the local authority to require
. medical examination of all persons entering common lodging-
houses and casual wards to see if they are suffering from
small-pox, and to offer a reward for prompt information of
the presence of the disease.
(iii.) That the local authority should have power to order the keeper
of a common lodging-house in which there has been small-
pox to refuse fresh admissions for such time as may be
required by the authority.
(iv.) That the local authority should be empowered to require the
temporary closing of any common lodging-house in which
small-pox has occurred.
(v.) That the local authority should have power to offer free’
lodgings to any inmate of a common lodging-house or casual
ward who may reasonably be suspected of being liable to
convey small-pox.
(vi.) That the sanitary authority should give notice to all adjoining
sanitary authorities of the occurrence of small-pox in
common lodging-houses or casual wards.
(vii.) That where the disease occurs, the Public Vaccinator or the
Medical Officer of Health should attend and vaccinate the
inmates of such lodging-houses or wards, except such as
should be unwilling to submit themselves to the operation.
Relaxation of the Vaccination Law.
After careful consideration and much study of the subject, we have
arrived at the conclusion that it would conduce to increased vaccination
if a scheme could be devised which would preclude the attempt (so often
a vain one) to compel those who are honestly opposed to the practice to
submit their children to vaccination, and, at the same time, leave the law
‘to operate, as at present, to prevent children remaining unvaccinated
owing to the neglect or indifference of the parent. When we speak of
an honest opposition to the practice, we intend to confine our remarks to
cases in which the objection is to the operation itself, and to exclude
REPORT OF THE ROYAL VACCINATION COMMISSION. 699
cases in which the objection arises merely from an indisposition to incur
the trouble involved. We do not think such a scheme impossible.
‘i Tt must of course be a necessary condition of a scheme of this
escription that it should be such as would prevent an objection to the
practice being alleged merely as an excuse to save the trouble connected.
with the vaccination of the child. We may give the following as
examples of the methods which might be adopted. It might be provided
that if a parent attended before the local authority and satisfied them
that he entertained such an objection, no proceedings should be taken
against him. Or, again, a statutory declaration to that effect before any
one now authorised to take such declaration, or some other specified
official or officials, might be made a bar to proceedings. We do not think
it would be any real gain to parents who had no conviction that the
vaccination of their children was calculated to do mischief, to take either
of these steps rather than submit them to the operation.
It is in England that the point we have been recently discussing is of
most practical importance, but if our suggestion were adopted the change
should, of course, be made in all parts of the United Kingdom.
(Signed) HERSCHELL.
JAMES PAGET.
CHARLES DALRYMPLE.
W. GUYER HUNTER.
EDWIN H. GALSWORTHY.
JOHN 8. DUGDALE.
M. FOSTER.
JONATHAN HUTCHINSON.
FREDERICK MEADOWS WHITE.
SAM. WHITBREAD.
JOHN A. BRIGHT.
Bret Incr,
August 1896. Secretary.
The undersigned do not find themselves able to go so far in recom-
mending relaxation of the law as is implied. We think that in all cases.
in which a parent or guardian refuses to allow vaccination, the person so-
refusing should be summoned before a magistrate, as at present, and that
the only change made should be to permit the magistrate to accept a
sworn deposition of conscientious objection, and to abstain from the
infliction of a fine. '
We are also of opinion that, in spite of the difficulties as set forth in
paragraph 533,* a second vaccination at the age of twelve ought to be made
compulsory.
W. GUYER HUNTER.
JONATHAN HUTCHINSON.
* Of the Final Report.
700 SUPPLEMENTARY APPENDIX.
‘We the undersigned desire to express our dissent from the proposal to
retain in any form compulsory vaccination.
We cordially concur in the recommendation that conscientious ob-
jection to vaccination should be respected. The objection that mere
negligence or unwillingness on the part of parents to take trouble might
keep many children from being vaccinated would be largely, if not wholly,
removed by the adoption of the Scotch system of offering vaccination at
the home of the child, and by providing for medical treatment of any
untoward results which may arise. 4
We therefore think that the modified form of compulsion recommended
by our colleagues is unnecessary, and that in practice it could not be
carried out.
The hostility which compulsion has evoked in the past toward the
practice of vaccination is fully acknowledged in the Report. In our
opinion the retention of compulsion in any form will in the future cause
irritation and hostility of the same kind.
The right: of the parent on grounds of conscience to refuse vaccination
for his child being conceded, and the offer of vaccination under improved
conditions being made at the home of the child, it would in our opinion be
best to leave the parent free to accept or reject this offer.
SAM. WHITBREAD.
JOHN A. BRIGHT,
W. J. COLLINS.
J. ALLANSON PICTON.
Note.—Dr. Collins and Mr. Picton sign the abure note of reservation,
though they have not signed the Report, A statement of their grounds of
dissent from the Report will be found in the form of an Appendie (65 pages)
to the Report. They make the following recommendations :—
In accordance with the sub-head No. 2 of the reference to the
Commission, we would suggest the following as the means other than
vaccination which should be employed for protection of a community
from small-pox :—
1. Prompt notification of any illness suspected to be small-pox.
Improved instruction in the diagnosis of small-pox.
2. A hospital, suitably isolated, of adequate accommodation, in per-
manent readiness, and capable of extension if required. No
other disease to be treated at the same time in the same
place.
3. A vigilant sanitary staff ready to: deal promptly with first cases,
and if necessary to make a house-to-house inspection. The
medical officer of health to receive such remuneration as to
render him independent of private practice.
4. Prompt removal to hospital by special ambulance of all cases
which cannot be properly isolated at home. Telephonic com-
munication between Health Office and hospital.
REPORT OF THE ROYAL VACCINATION COMMISSION, 701
5. Destruction of infected clothing and bedding, and thorough dis-
infection of room or house immediately after removal of the
patient. :
. Daily observation (including, where possible, taking the tempera-
ture and inspection for rash) of all persons who have been in
close contact with the patient during his illness ; such super-
vision to be carried out either in quarantine stations (away
from the hospital) or at their own homes.
. Closure of schools on the occasion of the occurrence of small-pox
among the scholars or teachers.
8. Hospitals and quarantine stations to be comfortable and attractive,
and so administered as to secure the confidence of the public.
Hospital treatment to be free to all classes, and compensation
to be paid to those detained or otherwise inconvenienced in the
public interest, at the public expense.
9. Tramps entering casual wards to be medically inspected, their
clothing to be disinfected, and bath provided. The measures
for detection and isolation of small-pox in common lodging-
houses suggested in section 507 of the Report to be carried
out.
10. International notification of the presence of small-pox, and
special vigilance at seaports in communication with infected
places, after the plan adopted in the case of cholera.
11. Attention to general sanitation—prevention of overcrowding.
abundant water supply, and frequent removal of refuse.
a
4
They conclude as follows :—
We believe the methods of isolation of the infected, disinfection, and
the observance of strict cleanliness, are both more successful and more
legitimate methods for the State to encourage. They have the advantage
of applying the preventive only where it is required ; and they do not
necessitate an operation upon the person of every healthy individual.
We therefore recommend that the law be amended by the repeal of
the compulsory clauses of the Vaccination Acts. But in consideration
of the prevalent belief in the value of vaccination as a prophylactic for
an indefinite period, we suggest that in other respects the law should be
left as it is, subject, however, to such modifications as are recommended
for the diminution of attendant risks. The precedent established in the
case of the abolition of compulsory church rates might be followed with
advantage. In that case all machinery for laying and collecting the rate
was left intact though the power of enforcement was taken away. The
effect of our recommendation, if adopted, would be that vaccination
would continue to be provided as at present for those who desire to avail
themselves of it, but efforts to secure vaccination would be limited to
moral influence—in a word, the whole country would be in the position of
those unions in which the guardians have abandoned compulsion. 55 of
The grounds on which we object to the enforcement of vaccination
by penalties necessarily lead us to object to any method of indirect
702 SUPPLEMENTARY APPENDIX.
compulsion. We regard as both inexpedient and unjust exclusion from
any branch of the public service because of the refusal to submit to
vaccination or re-vaccination. The injustice is perhaps most severely felt
in the case of candidates for employment as pupil-teachers in public
elementary schools. There are now districts in which, owing to the
general opposition to vaccination, scarcely a girl or boy can be found who
is legally eligible, and candidates have to be brought in at great incon-
venience from surrounding districts. The existence of an exceptional
case ov cases in which such rejected candidates have at some time after-
wards taken small-pox is in our view no justification for the continuation
of this grievance. Statistics furnished to the Commission prove that
large numbers of vaccinated or re-vaccinated persons have taken the
disease ; and we are not aware of any evidence to show that vaccinated
pupil-teachers have any special immunity. If our recommendations were
carried out, the danger of contagion would be greatly diminished in
schools, as elsewhere.
On the whole, then, while there is much in the report of our colleagues
from which we dissent, and we have accordingly abstained with reluctance
from adding our signatures to theirs, we are at one with them in holding
that it is unwise to attempt to enforce vaccination on those who regard it
as useless and dangerous. We, however, go further, and agree with our
colleagues, Mr. Whitbread and Mr. Bright, that it would be simpler and
more logical to abolish compulsory vaccination altogether.
INDEX.
703
INDEX.
A
Abbé’s condenser, 74
Aberration, chromatic, 67
spherical, 67
Abscess, 173
Achorion Schonleinii, 584
Actinomycosis, 413—447
bovis, 434
— cattle to cattle, 444
cultivation of, 436
hominis, 431
—— in cattle, 429
—— in man, 426
——- transmission of, from man to
lower animals, 443
Agar-agar, 625
Air, compressed, 24
—— examination of, 140
— Hesse’s method, 141
— Koch’s >» Al
— Petri’s » 142
—— Pouchet’s ,, 143
--— Sedgwick’s ,, 143
Aitken’s tubes, 129, 630
Alexines, 57
Alum carmine, 616
Ameeba coli, 610
Anaerobes, classified, 494
cultivation of, 130—133
Aniline water, 617
Animals Order, 1878, 455
Anthrax, 42, 183, 191—216
—— bacillus of, 192, 197
—-— in horses, 207
—— in swine, 203
origin and spread of, 198
—— preventive inoculation, 209
_— »» Measures in, 199 ;
Anthrax, stamping-out system, 210
Antiseptics, 30
Antitoxins, 56, 57
—— diphtheria, 58—62
—— septic infections, 63
—— tetanus, 62
—— typhoid, 64
Arthrospores, 19
Artificial fluids as media, 120
Ascococcus, 14
--— Billrothii, 498
— citreus, 498
Ascomycetes, 584
Asiatic cholera, 360
Aspergillus albus, 588
—— clavatus, 588
—— flavescens, 588
—— flavus, 586
—— fumigatus, 586
—— glaucus, 586
-—— nidulans, 588
—— niger, 587
—— ochraceus, 588
—— repens, 586
— subfuscus, 588
B
Babés’ incubator, 633
Bacillus, 13, 488
—~— acidiformans, 498
—— acidi lactici, 498
—— aerogenes, 499
capsulatus, 499
——- aerophilus, 499
—— albus, 499
—— —— anaerobiescens, 499
—— — cadaveris, 499
—— —— putidus, 499
5 45
706
Bacillus allantoides, 499
---~ allii, 499
—— alvei, 470
—-- amylobacter, 502
—— amylozyma, 500
—— anaerobicus liquefaciens, 500
—— anthracis, 192
—— aquatilis, 500
— -—— fluorescens, 500
—— graveolens, 500
—— sulcatus, 500
arborescens, 500 —
argenteo-liquefaciens, 501
— phosphorescens, 401
aurantiacus, 501
aureus, 501
beroliniensis indicus, 502
—— brassice, 502
—— brevis, 502
——- brunneus, 502
——- buccalis fortuitus, 502
— —— maximus, 502
minutus, 502°
—- butyricus, 502, 503 «-
-—— cadaveris, 503 :
—— canalis capsulatus, 504
—— —— parvus, 504
—— candicans, 504
—— capsulatus, 504
.— —— mucosus, 504
suis, 504
-——— carabiformis, 504
—-— carnicolor, 504
--— carotarum, 505
—— cavicida, 505
——- chromo-aromaticus, 505
SS Havaniensis, 505
-—— circulans, 505
-— citreus cadaveris, 505
—— cloace, 505
—— coeruleus, 505
—— coli communis, 344
—— -— similis, 506
—— constrictus, 506
| —— coprogenes foetidus, 506
| —— —— parvus, 506
ih
|
‘.
*
iy
crassus aromaticus, 506
—— sputigenus, 506
cuniculicida, 228
cuticularis, 506
--~ albus, 506
la
| 7 cyaneo-fuscus, 507
Bacillus cyaneo-phosphorescens, 507
—— cyanogenus, 507, 508
— cystiformis, 508
—— delicatulus, 508
—— dentalis viridans, 508
—— dentriticus, 508
— devorans, 508
—— diffusus, 508
—— diphtheriz, 332—336
columbarum, 336
—-— —— vitulorum, 509
—— dysodes, 509 ~
--— endocarditidis capsulatus, 509 |
——- —— griseus, 509
—- enteritidis, 372 |
—— epidermidis, 509
—— erysipelatis suis, 356
erythrosporus, 509
—— figurans, 510, 511
— filiformis, 511
Havaniensis, 511
—— flavescens, 511
— flavocoriaceus, 511
—— fluorescens aureus, 511
—— —— liquefaciens, 511
—— -— longus, 512
— —— minutissimus, 512 —
— —- nivalis, 512
—— —— non-liquefaciens, 512
—— —— putidus, 512
———~ —- tenuis, 512
—— feetidus, 513
‘ozene, 513
—— fulvus, 513
-_— fuscus, 513
limbatus, 513
—— gallinarum, 513
—— gasoformans, 513
—— glaucus, 513
—— gliscrogenus, 513
—— gracilis, 514
— granulosus, 514
-—— graveolens, 514
— guttatus, 514
—— hemorrhagic septicemia, 231
—— halophilus, 514 :
— Hansenii, 514
—— Havaniensis, 505
—— —— liquefaciens, 514
-—— helvolus, 515
—— heminecrobiophilus, 515
—— hepaticus fortuitus, 515
!
INDEX,
Bacillus Hessii, 515
—-— hyacinthi septicus, 515
——— hyalinus, 515
—— hydrophilus fuscus, 515
—— ianthinus, 516
—— implexus, 516
—— in acne contagiosa in horses, 516
—— in cancer, 516
—— incanus, 518
-—— in cholera in ducks, 516
——-- in choleraic diarrhcea, 516
~-— indicus, 518
—— indigogenus, 519
-—- in diphtheritic disease of calves
516
—-- in erythema nodosum, 517
—— in infectious disease of bees, 471
inflatus, 519
in fc wl enteritis, 230
—— in gangrene, 517
—— in grouse disease, 517
—— in hog cholera, 517
—— in infantile diarrhoea, 517
-—— in intestinal diphtheriain rabbits,
517
—— in jequirity infusion, 517
in measles, 517
in noma, 517
—— in potato rot, 517
—— in purpura hemorrhagica, 517
in putrid bronchitis, 518
—— in “red-cod,” 518
——— in rhinoscleroma, 518
— — in saliva, 518
-.—— in swine fever, 346-352
—— —— erysipelas, 354
——- —— measles, 354
inunctus, 519
invisibilis, 518
— in whooping cough, 518
— iridescens, 519
— iris, 519
—- lactericeus, 520
— lactis aerogenes, 519
albus, 520°
= erythrogenes, 520
— — pituitosi, 520
—— leporis lethalis, 520
—— leprx, 407
—— leptosporus, 520
_—— limbatus acidi lactici, 520
——— limosus, 520
7
707
Bacillus liodermos, 521
—— liquefaciens, 521
—— —— communis, 521
—— -—— magnus, 521
—— —— parvus, 521
—— liquidus, 521
_—— litoralis, 521
—— lividus, 524
—— luteus, 524
-—— maidis, 524
—— mallei, 452
—— megatherium, 524
—— membranaceus amethystinus, 525
—— meningitidis purulente, 525
—— mesentericus fuscus, 525
-——— —— ruber, 525
—— —— vuilgatus, 526
—— multipediculus, 522
—— muscoides, 522
—— mycoides, 522
—— —— roseus, 522
~—— neapolitanus, 522
—— necrophorus, 523
— nitrificans, 523
nodosus parvus, 523
nubilus, 523
— ochraceus, 523
— cedematis aerobicus, 523
—— —— maligni, 220
—— of Belfanti and Pascarola, 523
— of Colomiatti, 526
—— of Fulles, 526
-~— of Guillebeau, 526
——- of Letzerich, 526
— of Martinez, 526
—— of Nocard, 526
— of Okada, 526
—— of quarter-evil, 217
—— of Roth, 526
—— of Sattler, 526
—— of Schaffer, 527
—— of Scheurlen, 527
—— of Schou, 527
of septicemia of buffaloes, 226
—— of guinea-pigs,224 —
-~—— —— of mice, 225
—— of swine plague, 351
— of Tommasoli, 527
—— of Utpadel, 527
—— of Winogradsky, 527
—— ophthalmia, 190
—— ovatus minutissimus, 527
708
Bacillus oxytocus perniciosus, 527
— pestifer, 528
—— phosphorescens gelidus, 528
—— —— indicus, 528
—— —— indigenus, 528
—— plicatus, 528
—— pneumonie croupose, 233
—— pneumosepticus, 528
—— polypiformis, 528
prodigiosus, 528
—— proteus fluorescens, 529
—— pseudo-diphtheriticus, 529
—— —— tuberculosis, 529
--— pulp pyogenes, 529
---— punctatus, 529
— putrificus coli, 529:
—— pyocyaneus, 529
-— pyogenes foetidus, 530
soli, 530
radiatus, 530
—— —— aquatilis, 530
—— ramosus, 531
—— reticularis, 531
—— rhinoscleroma, 411
— rosaceus metalloides, 531
—-— rubefaciens, 531
—— rubellus, 531
—— ruber, 531
—--~ rubescens, 532
~— rubidus, 5382 ‘
—— sanguinis typhi, 532
—— saprogenes, 532
—— scissus, 532
—— septicemiz hemorrhagice, 231
—— septicus, 532
—— —- acuminatus, 533
—— —— agrigenus, 533
—— -—— keratomolacie, 533
— — ulceris gangreenosi, 533
—— —— vesice, 533
——- sessilis, 533.
——— smaragdino-phosphoreseens, 533
-—— smaragdinus foetidus, 534
—— solidus, 534
—— spiniferus, 534
——- spinosus, 534
—-— stolonatus, 534
—— stoloniferus, 534
striatus albus, 534
—— —— flavus, 534
—— subflavus, 535
——- subtilis, 535
, INDEX.
Bacilius subtilis similans, 536
—— sulfureus, 536
—— superficialis, 537
-~---- syphilis, 410
—— tenuis sputigenus, 537
—— termo, 537
—-- tetani, 457
-—— thelassophilus, 537
—— thermophilus, 537
—— tremelloides, 537
—— tuberculosis, 378
gallinarum, 402
——— tumescens, 537
-—- typhi abdominalis, 342—346
— -— murium, 359
-— ubiquitus, 537
» —— ulna, 538
vacuolosis, 538
—— varicosus conjunctive, 538
——- venenosus, 538
brevis, 538
— —— invisibilis, 538
—— —— liquefaciens, 539
—— ventriculi, 539
—— vermicularis, 539
vermiculosus, 539
——- violaceus, 539
—— —— laurentius, 539 ©
virescens, 539
—— viridis pallescens, 539
—— viscosus, 540
—- Zurnianus, 540
Bacteriacez, 480
Bacteria, chemical action on, 25
products of, 39
— composition of, 11
distribution of, 29
form of, 13
growth of, 23, 25
—— in cattle plague, 296
in diphtheria of pigeons, 336
—— in distemper, 358
—— in foot and mouth disease, 300
—— in horse-pox, 312
—— in liquids, 83
— in louping-ill, 463
—— in measles, 283
in pus, 176
—— in rabies, 460
— in scarlet fever, 262
—— in sheep-pox, 298
—— in small-pox, 285
INDEX.
Bacteria in vaccine lymph, 324—826
-—— in yellow fever, 260
—— microscopic examination of, 83
—— nitrifying, 27
—— pathogenic, 27
-~— saprogenic, 26
stained, 85
-— unstained, 84 '
Bacterium aerogenes, 540
— brunneum, 540 |
——- fusiforme, 540 |
—— gingivee pyogenes, 540
— hyacinthi, 540
—— hydrosulfureum ponticum, 540
—— litoreum, 540
— luteum, 540
—— merismopedioides, 541
navicula, 541
—— photometricum, 541
—— synxanthum, 541
—-— termo, 541
—**holeideum, 541
urex, 541
-—— violaceum, 541
-— Zopfii, 542
Beggiatoa alba, 542
roseo-persicina, 542
Bibliography, 639
Bilious fever, 183
Biondi’s stain, 615
Bismarck-browa, 617
Blepharadenitis, 184
Blood-serum, 113, 114, 119
Borax carmine, 617
Botrytis Bassiana, 588
Bread paste, 118
Broncho-pneumonia, 183
Broth, 118
Bunge’s method of staining flagella,
92
C
Camera lucida, 621
Cancer bodies, 610
Caoutchouc caps, 626
Caterpillars, disease of, 492
Cattle plague, 293—296
—— tuberculosis of, 389
Cerebro-spinal meningitis, 184
Chemical action on bacteria, 25
__— disinfectants, 32—36
__— products of bacteria, 39
709
Chemiotaxis, 54
Chionyphe Carteri, 588
Cholera, 360
—— bacteria of, 371
—— comma bacilli of, 361—367
—— diarrheea from meat-poisoning.
371
—— diarrheea in fowls, 373
—— nostras, 370
—— protective inoculation, 369
—— Ptomaines, 41
—— spirillum of, 361
Chromogenic bacteria, 25
Cladothricez, 480
Cladothrix, 479
—— dichotoma, 543
—— Forsteri, 544
— intricata, 544
—— invulnerabilis, 544
Classification, 475, 487, 482, 481
Clostridium butyricum, 5
— feetidum, 545
Coccacere, 480
Cocci, 485
Coccidia, 609
Comma bacilli, 361—367
Cover-glass preparations, 86
Cow-pox, 274—282, 312—324, 326—
328
Crenothrix Kiihniana, 545
D
Dacryocystis, 184
Damp chambers, 628
D’Arsonval’s incubator, 631
Davaine's septicemia, 228
De Bary’s classification, 481
Decalcifying, 93, 615
Defensive proteids, 57
Deneke’s comma bacillus, 367
Desiccators, 638
Diarrhea, choleraic, 371
Diphtheria, 330, 182
——- antitoxin in, 55—62
-— bacillus of, 332
—— in milk, 338
—— in pigeons, 336
—— Lifiler’s stain for, 88
—— Ptomaines of, 46
Diplococeus, 14
—— albicans amplus, 547
___.. —— tardissimus, 547
710
Diplococcus
547
citreus conglomeratus,
liquefaciens, 547
—— coryze, 547
—— flavus liquefaciens tardus, 547
—— fluorescens foetidus, 547
-—- intercellularis meningitidis, 548
—- luteus, 548
—— pneumoniz, 233—238
-—— -—— of horses, 548
—-— roseus, 548
~—— subflavus, 548
Distemper in dogs, 358
Drop-cultures, 120
Duck cholera, 230
Dysentery, 373
E
Ebner’s solution, 615
Egyptian ophthalmia, 190
Ebrlich’s staining method, 89
Electricity, 24, 127
Embedding, 93, 615
Empusa muscz, 581
—— radicans, 582
Endospores, 18
Enteric, 41, 340
Enzymes, 48
Eosin, 89,617
Epidemic disease of deer and boars,
227
—— —— of ferrets, 358
—— —— of mice, 358
Erysipelas, 185—189
Esmarch’s roll-cultures, 113
Experiments on animals, 134, 394
F
Farrant’s solution, 621
Favus, 584
Filter, hot water, 624
Flacherie, 472
Flagella, i7, 90
Fliigge’s classification, 481
Foot rot, 464
Form of bacteria, 13
Foul-brood, 469
Fowl cholera, 228
-—- enteritis, 230
— scab, 586
Friedlinder’s bacillus, 233
Fuchsine, 617
INDEX.
G
Gangrene, 182
Gas chamber, 127
Gases produced by bacteria, 24
Gentian violet stain, 617
Giant cells, 376
Gibbes’ solution, 618
Glanders, 451
— bacillus of, 452
—— mallein in, 454
—— ptomaine, 48
Glycerine agar, 104
Glycerine gelatine, 615
Gonococcus of Neisser, 189
Gonorrheea, 189
Gram’s method, 88, 89, 97, 618
Grease, 303
Grouse disease, 230
H
Hematococcus bovis, 548
Hematomonas carassii, 605
——- cobitis, 603
Hematoxylin, 618
Hamatozoa, 589, 593, 599, 603
Hardening, 93, 615
Helicobacterium aerogenes, 549
Herpetomonas Lewisi, 599
Hessert’s stain, 92
Historical introduction, 1
Horse-pox, 303, 312
Hot air and steam, 36, 623
Hot water filter, 624
Hueppe’s classification, 482
—— inspissator, 630
Hydrophobia, 459
Hyphomycetes, 581
Hypodermii, 581
I
Tllumination, 75
Immersion system, 69
Immunity, 50—57
Impression preparations, 92
Incubators, 631
Infectious pleuro-pneumonia, 239, 247 ,
Influenza, 247—249
Tnspissators, 629
Involution forms, 15
Iodine, 618
Isolation of micro-organisms, 139
Israel’s case, 628
K
Klein’s micrococcus of pneumonia, 238
Kleinenberg’s solution, 616
Koch’s comma bacillus, 361
—— postulates, 9
—— serum steriliser, 629
— steam steriliser, 622
L
Leprosy, 406
-—— bacillus of, 407
—— stamping out system, 409
Leptothrix, 480
— buccalis, 549
— gigantea, 549
Leuconostoc, 549
Leukemia, 184
Light, effect of, on bacteria, 24
Lister’s flasks, 128, 630
Lithium carmine solution, 618
Loffler’s solution, 619
—— stain, 88, 90
Louping ill, 462
bacillus of, 463
Lutesch’s stain, 91
M
Madura diséase, 447
_ Magenta solution, 618
Malaria, 589
Malignant cedema, 220
Mallein, 454
Measles, bacteria in, 183, 283
Media, 99
Merismopedia, 14
Methyl violet, 619
Methylene blue, 618, 619
Micrococcus acidi lactici, 550
—— —— —— liquefaciens, 550
—— aerogenes, 551
—— agilis, 551
—_+ + eitreus, 551
—~— albus liquefaciens, 551
—— amylivorus, 551
—— aquatilis, 551
—— —-— invisibilis, 551
—— auranticaus, 551
—- botyogenus, 552
-—— candicans, 552
— candidus, 552
carneus, 552
cerasinus siccus, 552
INDEX.
Micrococcus cereus albus, 178
—— — flavus, 178
-—— cinnabareus, 552
—— citreus, 552
—— concentricus, 553
—— cremoides, 553
—— crepusculum, 553
-——— cumulatus tenuis, 553
—— endocarditidis rugatus, 553
—— fervidosus, 553
—-— Finlayensis, 553
-——- flavus desideus, 553
—— —— liquefaciens, 554
—— —— tardigradus, 554
—— feetidus, 554
—— Freudenreichi, 554
——- fuscus, 554
—— gingive pyogenes, 554
—— gonorrhee, 190
—— Havaniensis, 555
—— in abscess in rabbits, 556
— in Biskra button, 555
—— in gangrenous mastitis in sheep,
555
—— in infectious pleuro-pneumonia,
555
—-— in influenza, 555
—— in pemphigus, 555-6
—-— in pneumonia, 236, 238, 556°
—— in pyzmia in rabbits, 556
-— in septicemia in rabbits, 556
—— in syphilis, 557
—— in trachoma, 190
— lactis viscosus, 557
_ —— luteus, 557
—— melitensis (Malta fever), 557
—— ochroleucus, 557
——— plumosus, 557
—-— pneumoniz croupose, 236
—— pyogenes tenuis, 557 :
— rosaceus, 558
--—— rosettaceus, 558
—— salivarus septicus, 558
—- stellatus, 588
—— tetragenus, 558
—— —— mobilis ventriculi, 558
--- —— subflavus, 558
—— —— versatilis, 558
— urez liquefaciens, 559
——— versicolor, 559
-—— violaceus, 559
—— viticulosus, 559
711
712
Micrometer, 80
Microscope, 65, 612 ~
Microsporon furfur, 586
Microtomes, 94,614
Miescher’s tubes, 609
Mildew, 581
Milk, 120
—— scarlatina, 265, 282
—— tubercular, 291
Miquel’s bulbs, 129
Moist chambers and cells, 121
Moitessier’s gas regulator, 634
Monads, 601
Monas Okenii, 560
—— Vinosa, 560
—— Warmingii, 560
Mouse favus, 586
Movements of bacteria, 15, 24
Mucors, 583
Miiller’s fluid, 616
Myconostoc gregarium, 560
Myco-protein, 11
N
Nature of soil for bacteria, 23
Neelsen’s solution, 89, 98, 619
Nicols’ stain, 92
Nitromonas of Winogradsky, 560
Nutrient agar, 103
Nutrient gelatine, 100
ie)
Oidium albicans, 586
— lactis, 584
—— Tuckeri, 584
Oil immersion, 70
Orseille, 619
P
Parietti’s method, 148
Pasteur’s apparatus, 130
Pathogenic bacteria, 27
Pébrine, 471
Pediococcus acidi lactici, 560
cerevisize, 560
Penicillium glaucum, 588
Peronospora infestans, 582
Petri’s method exam. of air, 142
Pfeiffer bodies, 610
Phagocytosis, 27, 55
Photogenic bacteria, 25
Photography of bacteria, 150, 168
INDEX.
Photo-micrographic apparatus, 621.
Phycomycetes, 582
Phylaxins, 57
Picrie acid, 619
Picro-carmine, 619
Picro-lithium carmine, 620
Pilobolus, 583
Plague, bacillus of, 250, 252
Plate cultivations, 106
Platinum needles, 83, 616
Pleuro-pneumonia, 239
Pneumobacillus liquefaciens
242, 560
Pneumonia, 233—238
—— ptomaines, 48
Potash solution, 620
. Potato medium, 115—117
Pouchet’s aeroscope, 143
Preservation of preparations, 93°
Protective inoculation, 49
—— —— in anthrax, 209
in cholera, 369
—— —— in rabies, 460
—— —— in sheep-pox, 298
—— —— in small-pox, 285, 293
—— in swine erysipelas, 336-
Proteus capsulatus septicus, 560
hominis capsulatus, 224
—— in gangrene of the lung, 560
— wmicrosepticus, 560
—— mirabilis, 561
—— septicus, 561
—— sulfureus, 561
—- vulgaris, 561
—— Zenkeri, 562
Pseudo-diphtheritic bacillus, 335
Pseudo-diplococcus pneumoniz, 562:
Psorosperms, 609
Ptomaines, 39
Pus, bacteria in, 176
Putrefactive bacteria, 26, 28
Pyemia, 175
Q
Quarter-evil, 217
R
Rabies, 459—462
Rag-pickers’ septicaemia, 224
Refraction, 65
Relapsing fever, ‘257
Reproduction, 18
bovis,,
INDEX,
Respiration of bacteria, 22
Rhabdomonas rosea, 562
BRhinoscleroma, 411
Ringworm, 585
8
Saccharomyces acidi lactici, 580
— albicans, 579
—— anomalus, 579
—— apiculatus, 578
—— aquifolii, 580
—— cerevisiz, 577
—— conglomeratus, 578
—— ellipsoideus, 577
—— exiguus, 578
—— glutinus, 579
—— Hansenii, 580
-—— ilicis, 580
— Jorgensenii, 578
—— Ludwigii, 580
-—— Marxianus, 580
—— membranefaciens, 580
—— minar, 580
niger, 580
—— pastorianus, 578
-— pyriformis, 579
rosaceus, 580
sphericus, 579
Safranine, 620
Sapremia, 175
Saprogenic bacteria, 26
Saprolegina, 582
Saprophytic bacteria, 27
Sarcinz, 487
Sarcina alba, 562
—— aurantiaca, 562
-—— candida, 563
——~ flava, 563
—— hyalina, 563
—— intestinalis, 563
--— litoralis, 563
——- lutea, 563
-—- mobilis, 563
—— pulmonum, 563
—-~ Reitenbachii, 564
rosea, 564
— urine, 564
—- ventriculi, 564
Scarlet fever, 261
Schizomycetes, 477
Schlosing’s membrane regulator, 632
Sclavo’s stain, 91
Septiczemia, 175
—— of calves, 227
—— of Davaine, 224
—— of guinea-pigs, 224
—— of mice, 225
—-— of rabbits, 228
—— of rag-pickers, 224
Serum therapy, 56
Sheep-pox, 297
Small-pox, 182, 284-293, 326
Smut, 581
Soil, examination of, 144
Sozins, 57
Spherotilus natans, 564
Spirilla, 495
Spirillum amyliferum, 564
—— anserum, 564
—— attenuatum, 564
———— aureum, 564
— cholere Asiaticz, 361
—— choleroides, 565
— concentricum, 565
—— dentium, 565
—— Finkler and Prior, 258. -
— flavescens, 565
— — flavum, 565
— Giinther, 566
— leucomelaneum, 565.
—— lingue, 565
— marinum, 565
—— Metchnikovi, 373
—— Miller, 566
—— nasale, 566
—— Neisser, 566
—— Obermeieri, 258
—— plicatile, 466
—— Renon, 566
—— rosaceum, 566
—— Rosenbergii, 566
—— rubrum, 569
—— rufum, 566
—— rugula, 567
—— sanguineum, 567
—— saprophiles, 567
—— serpens, 568
—— Smith, 566
—— sputigenum, 568
—— suis, 568
—— tenue, 568
—— tyrogenum, 568
— undula, 568
— volutans, 568
713:
714
Spirillum Weibel, 566
Spirocheetz, 596
Spiromonas Cohnii, 569
volubilis, 569
Spontaneous generation, 3
Spores, 18
Staining spores, 90
Staphylococcus, 14
—— cereus albus, 178, 324
flavus, 178
—— pyogenes albus, 178
—— —— aureus, 176, 324
—— —— citreus, 178
—— pyosepticus, 569
—— salivarius pyogenes, 569
—— viridis flavescens, 569
‘Stémberg’s bulbs, 128, 630
‘Streptococcus, 14
—— acidi lactici, 569
—— albus, 569
-—— bombycis, 472
—— brevis, 569
—— cadaveris, 570
—— coli gracilis, 570
—— conglomeratus, 570
—— flavus desidens, 570 |
-—— giganteus urethre, 570
—— Havaniensis, 570
—— in contagious mammitis in cows,
570
571
in strangles, 571
—— liquefaciens, 571
—— mirabilis, 571
-—~— of Bonome, 571
—— of erysipelas, 185, 189
—— of Manneberg, 571
—— perniciosus psittacorum, 572
—— pyogenes, 178-184
radiatus, 572
——- septicus, 572
liquefaciens, 572
vermiformis, 572
‘Streptothrix, 496
—— actinomycotica, 431
—— albus, 572
——- asteroides, 573
—— aurantiaca, 573
carnea, 573
—— chromogena, 573
—- farcinica, 573
in progressive necrosis in mice, -
INDEX.
Streptothrix Forsteri, 573
-—~ Hoffmanni, 573
— liquefaciens, 573
—— Madure, 449
—— musculorum suis, 573
— odorifera, 573
——- violacea, 573
Substage condenser, 74
Suppuration, 48
Surgical fever, 182
Surra, 593
Swine erysipelas, 354
——- fever, 46, 227
—— measles, 364
Syphilis, 183, 410
T
Tarichium megaspermum, 582
Temperature for bacteria, 24
Test-tube cultivations, 105
Tetanus, 457
ptomaine, 41
Thermo-regulators, 635
Tilletia caries, 581
Toxalbumins, 39
Trachoma, 412
Trenkmann’s stain, 91
Trichomonas, 607
Tricophyton tonsurans, 585
Tripod levelling apparatus, 627
Tuberculosis, 43, 375
—— and meat, 397
and public health, 391
—— antitoxin, 64
— bacillus of, 378
—— giant cells in, 376
—~- in animals, 389, 394, 399, 400
—— in man, 387
Typhoid, 340
Typhus, 259
U
Ulcerative endocarditis, 183
Undulina ranarum, 607
Uredo, 581
Urine as a medium, 120
Urobacillus Duclauxi, 573
Freudenreichi, 574
—— Maddoxi, 574
-—— Pasteuri, 574
—— Schutzenbergi, 574
INDEX. 715
Urocystis occulta, 581
Weigert’s stain, 296
Ustilago carbo, 581 Wort gelatine, 104
v x
Van Ermengen’s stain, 91 Xylol, 616
Vegetable infusions as media, 120
Vesuvin, 620 Y
Vibrio rugula, 574 Yeasts, 577
Yellow fever, 182, 259
WwW
Warmstage, 123, 124 Z
Water bath, 624 Zooglea, 13
—— examination of, 145 Zopf’s classification, 480
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