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Annals of Tropical Medicine
and Parasitology
THE UNIVERSITY OF LIVERPOOL
Annals
OF
Tropical Medicine and
Parasitology
ISSUED liY THE
Liverpool School of Tropical Medicine
Edited by
Professor J. W. W. STEPHENS, M.D.Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
AND
Professor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM
volume VII
(March 31, to December 30, 1913)
fVith forty- two plates., twenty• two figures in text, and seventeen charts
LIVERPOOL:
AT THE UNIVERSITY PRESS, 57 ASHTON STREET.
HAP ft ' -J (;*.r.v r . , ;
1 '■ * ' " ^ '* -
Editorial Secretary
Ur. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool .
CONTENTS
No. i. March 31, 1913
PAGE
Macfie, J. W. Scon.
Trypanosomiasis of Domestic Animals in Northern Nigeria. Plates I—III ... l
Stephens, J. W. W. ; and Fantham, H. B.
Further Measurements of Trypanosoma rhodesicnse and T. gambiensc ... 27
Mayer, T. F. G.
A New Mosquito-proof and Storm-proof House for the Tropics. Plate IV ... 41
Christophers, Major S. R.
Contributions to the Study of Colour Marking and other Variable Characters
of Anophelinae with Special Reference to the Systematic and Phylogenetic
Grouping of Species. Plates V—VIII . 45
Black lock, B.
A Study of the Posterior Nuclear Forms of Trypanosoma rhodesiense (Stephens
and Fantham) in Rats ... ... ... ... ... ... ... ... 101
Balfour, Andrew.
Animal Trypanosomiasis in the Lado (Western Mongalla) and Notes on Tsetse
Fly Traps and on an Alleged Immune Breed of Cattle in Southern
Kordofan. Plates IX, X . 113
Thomson, David.
Sanitation on the Panama Canal Zone, Trinidad and British Guiana. Plates
XI, XII, XIII. 125
Thomson, John Gordon ; and Thomson, David.
The Cultivation of One Generation of Benign Tertian Malarial Parasites
(Plasmodium viva.v) in Vitro, by Bass’s Method. Plate XIV . 153
Scon, H. Harold.
Fulminating Cerebro-Spinal Meningitis in Jamaica ... ... ... ... 165
CONTENTS
No. 2. June io, 1913
PACE
Kinghorn. Allan ; Yorkf, Warrington ; and Lloyd, Llewellyn.
Final Report of the Luangwa Sleeping Sickness Commission of the British
South Africa Company, 1911-1912. Plate9 XV—XXVI. 183
Stephens, J. W. W. ; and Blacklock, B.
On the Non-identity of Trypanosoma brucei (Plimmer and Bradford, 1899)
with the Trypanosome of the same name from the Uganda Ox. 303
Todd, John L.
Concerning the Sex and Age of Africans Suffering from Trypanosomiasis ... 309
Darling, S. T.
The Identification of the Pathogenic Lntamoeba of Panama ... . 321
Newstead, Professor R.
A New Tsetse Flv from the Congo Free State ; and the Occurrence of GLossina
austeni in German Fast Africa ... . . 331
Marsdf.n, Prosper H.
Examination of the Root of an /pomvea from Rhodesia. Plate XXV II ... 335
No. 3A. August 11, 1913
Macfie, J. \\ . Scott.
On the Morphology of the Trypanosome ( T . nigrritnst\ n. sp.) from a Case of
Sleeping Sickness from Eket, Southern Nigeria. Plate XXVIII ... 339
No. 3B. November 7, 1913
Macfie, J. W. Scott. !
Preliminary Note on the Development of a Human Trypanosome in the Gut
of Stomoxys nigra . ... ... ... . ... 359
Rogers, YV.
A Note on a Case of Lou loa 363
Barratt, J. O. YVakelin.
Recent Experimental Research Bearing upon Blackwater Fever ... ... 367
Ross, G. A. Park
A Fictitious Native Disease (/ sigu't'bt'dhhi) ... ... ... ... ... 371
Seideiin, FIarald.
On ‘ Vomiting Sickness ' in Jamaica. Plate* XX 1 X-XX 1 II ... ... 377
CONTENTS
No. 4. December 30, 1913
PAGE
Stephens, J. \V. VY.
Studies in Blackwater Fever—(i) Statistical . . 479
Thomson, John Gordon ; and Thomson, David.
The Growth and Sporulation of the Benign and Malignant 'Tertian Malarial
Parasites in the Culture Tube and in the Human Host. Plates XXXIV,
XXXV. 509
Chalmers, Albert J. ; and O’Farrell, Captain \V. R.
The Trichonocardiases. Plates XXXVI—XXXVII . 525
Chalmers, Albert J. ; and Stirling, Captain A. D.
Epidemic Trichonocardiasis 541
OTarrell, Captain W. R.
Hereditary Infection, with Special Reference to its Occurrence in HyaJomma
aegyptium infected with Critkidia hyalommae. Plates XXXVIII—XL... 545
Black lock, B. : and Yorke, Warrington.
Trvpatiosoma vbax in Rabbits ... ... ... ... ... ... ... 563
Fantham, H. B. ; and Porter, Annie.
The Pathogenicity of Nosnna apis to Insects other than Hive Bees. 569
Carter, Henry F.
On Certain Mosquitos of the Genera Banksinella , Theobald, and 7 aeniorhynchus,
Arribalzaga . 581
Theobald, Fred. V.
New Culicidae from the Sudan ... ... ... ... ... ... ... 591
Blxcklock. B. ; and Yorke, Warrington.
The Probable Identity of Trypanosoma congolense (Broden) and T. natinm
(Laveran) ... ... ... ... ... ... ... ... ... ... 603
Fantham, H. B. ; and Porter. Annie.
Herpetomonas straliomyiae, n. sp., a Flagellate Parasite of the Flies, Stratiomyia
ehamelt'on and S. pot am id a , with Remarks on the Biology of the Hosts.
Plate XLI ... ... ... ... ... ... ... ... 609
Thomson, J. G. ; and Fantham, H. B.
T of Babesia ( Piroplasma) canis in vitro. Plate XL 1 I
621
Volume VII
March, 1913
No. 1.
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Editor
Professor Sir RONALD ROSS, K.C.B., F.R.S., Major I.M.S. (Ret.),
M.D., D.P.H., F.R.C.S., D.Sc., LL.D.
In Collaboration with
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
Professor J. L. TODD, B.A., M.D., C.M. McGill, D.Sc. Liv.
H. WOLFERSTAN THOMAS, M.D., C.M. McGill.
ANTON BREINL, M.U.Dr.
H. B. FANTHAM, D.Sc.Lond., B.A. Cantab., A.R.C.S., F.Z.S.
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool.
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THE INCORPORATED
LIVERPOOL SCHOOL OF TROPICAL MEDICINE
{Affiliated with the University of Liverpool)
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Hon. Vice-Presidents: The Duke of Northumberland, K.G.
The Earl of Derby, G.C.V.O.
Earl Cromer, G.C.B.
Viscount Milner, G.C.B.
Lord Pirrie, K.P.
Sir Owen Philipps, K.C.M.G.
Sir Edwin Durning Lawrence, Bart.
Mr. O. Harrison Williams
COMMITTEE
Chairman: SIR WILLIAM H. LEVER, Bart.
Vice-Chairman: Mr. F. C. DANSON
Mr. H. J. Read, C.M.G.
Sir Alfred Dale
Sir W. B. Bowring, Bart.
Professor CATON
Professor SHERRINGTON
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Dr. C. J. Macalister
Mr. George P. Newbolt
Mr. C. Booth (Jun.)
Mr. T. F. Harrison
Mr. A R. Marshall
Mr. W. Roberts
Mr. Charles Livingston
Mr. George Brocklehurst
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Mr. J. A. Tinne
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B io Exchange Buildings, Liverpool
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University of Liverpool
| Council of University of Liverpool
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Staff
1. At the University of Liverpool
Professors . - JOHN WILLIAM WATSON STEPHENS, M.D.,
Cantab., D.P.H. Sir Alfred Jones Professor of
Tropical Medicine
ROBERT NEWSTEAD, M.Sc., J.P., F.R.S.,
A.L.S., F.E.S., Dutton Memorial Professor of
Entomology
Major Sir RONALD ROSS, K.C.B., F.R.C.S.,
D.P.H., M.D., D.Sc., LL.D., F.R.S., Nobel
Laureate 1902. (Indian Medical Service, retired).
Professor of Tropical Sanitation
Assistant Lecturers - HAROLD BENJAMIN FANTHAM, D.Sc., B.A.,
Assistant Lecturer in Parasitology
HAROLD WOLFERSTAN THOMAS, M.D.,
C.M.
Assistant Lecturer
in Entomology HENRY FRANCIS CARTER, S.E.A.C. Dipl.,
f.e.s.
Honor at y Lecturers - J. O. WAKELIN BARRATT, M.D., D.Sc., (Lond.)
Major JOSEPH FITZGERALD BLOOD, M.D.,
M.Ch. (Indian Medical Service, retired) '
Prof. ERNEST EDWARD GLYNN, M.A., M.D.,
(Cantab.), M.R.C.P., M.R.C.S.
Prof. E. W. HOPE, M.D., D.Sc.
WILLIAM THOMAS PROUT, M.B., C.M.G.
Assistant in
Cryotherapy JOHN GORDON THOMSON, M.A., M.B., Ch.B
Honorary Statistician WALTER STOTT
Bibliographer - - WALTER DRAWZ
Clerk to the
Laboratory - NORA MURPHY
2. At the Runcorn Research Laboratories
Director - - - WARRINGTON YORKE, M.D.
Assistant - - - B. BLACKLOCK, M.D., D.P.H.
3. At the Royal Southern Hospital, Liverpool
Physicians - - CHARLES JOHN MACALISTER, M.D.,
F.R.C.P.
JOHN LLOYD ROBERTS, M.D., M.R.C.P.
Surgeons - - . D. DOUGLAS CRAWFORD, F.R.C.S.
ROBERT JONES, F.R.C.S.
GEORGE PALMERSTON NEWBOLT, M.B.,
F.R.C.S.
Tropical Pathologist - JOHN WILLIAM WATSON STEPHENS, M.D.,
Cantab.
Clinical Pathological
Assistant - DAVID THOMSON, M.B. Ch.B., D.P.H.
4. On Expedition
HAROLD WOLFERSTAN THOMAS, M.D.,
C.M.
HARALD SEIDELIN, M.D.
Laboratory
JOHNSTON LABORATORY, UNIVERSITY OF LIVERPOOL
Research Laboratory:
RUNCORN
Hospital:
ROYAL SOUTHERN HOSPITAL, LIVERPOOL
Secretary's Office:
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NOTICE
The following courses of instruction will be given by the
Liverpool School of Tropical Medicine during 1913 :—
Full Course begins 6 January. Advanced Course begins 2 June.
Diploma Examination, 7 April. Certificate Examination, 30 June.
Full Course begins 15 September.
Diploma Examination, 15 December.
The full Course of Instruction is open to all qualified medical men,
and the Examination to all students who have taken out this full
course.
Fee for the full Course of Instruction—Thirteen Guineas.
Fee for the Diploma Examination—Five Guineas.
Fee for the Short Course of Instruction—Four Guineas.
Fee for the use of a School microscope during one term—Ten
shillings and sixpence.
For prospectus and further information, application should be made
to the Dean of the Medical Faculty, University of Liverpool.
The following have obtained the Diploma in Tropical Medicine of
the University of Liverpool: —
Diploma in Tropical Medicine
Date of
Diploma
1904 Augustine, Henry Joshua
1904 Bennett, Arthur King
1904 Bruce, William James
1904 Byrne, John Scott
1904 Clayton, Thomas Morrison
1904 Dalziel, John McEwen
1904 Dee, Peter
1904 Greenidge, Oliver Campbell
1904 Hehir, Patrick
1904 Khan, Saiduzzafor
1904 Laurie, Robert
1904 Maclurkin, Alfred Robert
1904 McConnell, Robert Ernest
1904 Nicholson, James Edward
1904 Philipson, Nicholas
1904 Sharman, Eric Harding
1904 Thomson, Frank Wyville
1904 Walker, George Francis Clegg
1905 Anderson, Catherine Elmslie
1905 Brown, Alexander
1905 Caldwell, Thomas Cathcart
1905 Critien, Attilio
1905 Hooton, Alfred
1905 Hudson, Charles Tilson
1905 Illington, Edmund Moritz
Date oj
Diploma
1905 Macfarlane, Robert Maxwell
1905 Maddock, Edward Cecil Gordon
1905 Moore, James Jackson
1905 Nightingale, Samuel Shore
1905 Radcliffe, Percy Alexander Hurst
1905 Young, John Cameron
1906 Adie, Joseph Rosamond
1906 Arnold, Frank Arthur
1906 Bate, John Brabant
1906 Bennetts, Harold Graves
1906 Carter, Robert Markham
1906 Chisholm, James Alexander
1906 Clements, Robert William
1906 Dundas, James
1906 Faichnie, Norman
1906 Jeffreys, Herbert Castelman
1906 Mackenzie, Donald Francis
1906 Pailthorpe, Mary Elizabeth
1906 Palmer, Harold Thornbury
1906 Pearse, Albert
1906 Sampey, Alexander William
1906 Smithson, Arthur Ernest
1906 Taylor, Joseph van Someron
1906 Taylor, William Irwin
1906 Tynan, Edward Joseph
Date of
Diploma
1906 Watson, Cecil Francis
1906 Willcocks, Roger Durant
1906 Williamson, George Alexander
1907 Allan, Alexander Smith
1907 Allwood, James Aldred
1907 Bond, Ashton
1907 Branch, Stanley
1907 Collinson, Walter Julius
1907 Davey, John Bernard
1907 Donaldson, Anson Scott
1907 Fell, Matthew Henry Gregson
1907 Gann, Thomas William Francis
1907 Graham, James Drummond
1907 Hiscock, Robert Carroll
1907 Keane, Joseph Gerald
1907 Kennan, Richard Henry
1907 Kenrick, William Hamilton
1907 Le Fanu, George Ernest Hugh
1907 Mackey, Charles
1907 Maddox, Ralph Henry
1907 McCarthy, John McDonald
1907 Raikes, Cuthbert Taunton
1907 Ryan, Joseph Charles
1907 Vallance, Hugh
1908 Caverhill, Austin Mack
1908 Crawford, Gilbert Stewart
1908 Dalai, Kaikhusroo Kustomji
1908 Dansey-Browning, George
1908 Davidson, James
1908 Dickson, John Rhodes
1908 Dowdall, Arthur Melville
1908 Glover, Henry Joseph
1908 Greaves, Francis Wood
1908 Goodbody, Cecil Maurice
1908 Harrison, James Herbert Hugh
1908 Joshi, Lemuel Lucas
1908 Le Fanu, Cecil Vivian
1908 Luethgen, Carl Wilhelm Ludwig
1908 Mama, Jam shed Byramji
1908 McCay, Frederick William
1908 McLellan, Samuel Wilson
1908 Pearce, Charles Ross
1908 Schoorel, Alexander Frederik
1908 Smith, John Macgregor
1908 Stewart, George Edward
1908 Tate, Gerald William
1908 Whyte, Robert
1909 Abercrombie, Rudolph George
1909 Allin, John Richard Percy
1909 Armstrong, Edward Randolph
1909 Barrow, Harold Percy Waller
1909 Beatty, Guv
1909 Carr-White, Percy
1909 Cbevallier, Claude Lionel
1909 Clark, William Scott
1909 Cope, Ricardo
1909 Fleming, William
1909 Hanschell, Hother McCormick
1909 Hayward, William Davey
1909 Henry, Sydney Alexander
1909 Innes, Francis Alexander
Date of
Diploma
1909 Jackson, Arthur Frame
1909 Kaka, Sorabji Manekji
1909 McCabe-Dallas, Alfred Alexander
Donald
1909 Meldrum, William Percy
1909 Murphy, John Cullinan
1909 Samuel, Mysore Gnananandaraju
1909 Shroff, Kawasjee Byramjee
1909 Thornely, Michael Harris
1909 Turkhud, Violet Ackroyd
1909 Webb, William Spinks
1909 Yen, Fu-Chun
1910 Brabazon, Edward
1910 Castellino, Louis
1910 Caulcrick, James Akilade
1910 Dowden, Richard
1910 Haigh, William Edwin
1910 Hamilton, Henry Fleming
1910 Hefferman, William St. Michael
1910 Hipwell, Abraham
1910 Homer, Jonathan
1910 Houston, William Mitchell
1910 James, William Robert Wallace
1910 Johnstone, David Patrick
1910 Korke, Vishnu Tatyaji
1910 Macdonald, Angus Graham
1910 Macfie, John Win. Scott
1910 Manuk, Mack Walter
1910 Murison, Cecil Charles
1910 Nanavati, Kishavlal Balabhai
1910 Nauss, Ralph Welty
1910 Oakley, Philip Douglas
19to Pratt, Ishmael Charles
1910 Sabastian, Tbiruchelvam
1910 Shaw, Hugh Thomas
1910 Sieger, Edward Louis
1910 Sousa, Pascal John de
1910 Souza, Antonio Bernardo de
1910 Waterhouse, John Howard
1910 White, Maurice Forbes
1911 Blacklock, Breadalbane
1911 Brown, Frederick Forrest
1911 Chand, Diwan Jai
1911 Holmes, John Morgan
1911 levers, Charles Langley
1911 lies, Charles Cochrane
1911 Ingram, Alexander
1911 Kirkwood, Thomas
1911 Knowles, Benjamin
1911 Liddle, George Marcus Berkeley
1911 Lomas, Emanuel Kenworthy
1911 Mackarell, William Wright *
1911 MacKniglit, Dundas Simpson
1911 Mascarenhas, Joseph Victor
x911 Murray, Ronald Roderick
1911 Oluwole, Akidiya Ladapo
1911 Rao, Koka Ahobala
1911 Sinton, John Alexander
1911 Tarapurvalla, Byramji Sbavakshah
1911 Taylor, John Archibald
1911 Woods, William Medlicott
Date of
Diploma
1912 Aeria, Joseph Reginald
1912 Anderson, Edmund Litchfield
1912 Borle, James
1912 Bowie, John Tait
1912 Brassey, Laurence Perdval
1912 Christie, David
1912 Dillon, Henry de Courcy
1912 Dunn, Lillie Eleanor
1912 Hardwicke, Charles
1912 Jagose, Jamshed Rustomji
1912 Kochhar, Mela Ram
1912 McGusty, Victor William Tighe
1912 Milne, Arthur James
1912 Mitra, Manmatha Nath
Date of
Diploma
1912 Myles, Charles Duncan
1912 Pelly, Huntly Nevins
1912 Prasad, Bindeshwari
1912 Prentice, George
1912 Ross, Frank
1912 Russell, Alexander James
Hutchison
1912 Ruthven, Morton Wood
1912 Sandilands, John
1912 Seddon, Harold
1912 Smalley, James
1912 Strickland, Percy Charles
Hutchison
1912 Watson, William Russell
EDITORIAL NOTICE
By order of the Committee of the Incorporated Liverpool School
of Tropical Medicine, the series of the Reports of the School, which
had been issued since 1899, were followed, from January 1, 1907,
by the Annals of Tropical Medicine and Parasitology, of which this
is the first number of the seventh volume.
Altogether twenty-one Memoirs, besides other works, were
published by the School since 1899, and of these ten, containing 519
quarto or octavo pages and 95 plates and figures, were published
during the two years 1904 and 1905.
The Annals are issued by the Committee of the School, and will
contain all such matter as was formerly printed in the Reports—that
is to say, accounts of the various expeditions of the School and of the
scientific work done in its laboratories at the University of Liverpool
and at Runcorn. In addition, however, to School work, original
articles from outside on any subject connected with Tropical
Medicine and Hygiene, and Parasitology may be published if found
suitable (see notice on back of cover); so that, in all probability, not
less than four numbers of the Annals will be issued annually.
Each number will be brought out when material sufficient for it has
been accumulated.
TRYPANOSOMIASIS OF DOMESTIC
ANIMALS IN NORTHERN NIGERIA
BY
J. W. SCOTT MACFIE, M.A., M.B., Ch.B.
WEST AFRICAN MEDICAL STAFF
(Received for publication 19 December , 1912)
Plates I—III
Trypanosomiasis is very prevalent amongst domestic animals in
Northern Nigeria, and, at any rate in the south-western portions of
the Protectorate, accounts not only for a heavy mortality, but also
for serious financial losses, due to sickness, forced sales and the
slaughtering of stock in unsuitable markets. It is difficult to give
any adequate idea of the grave nature of the losses, but some
conception of their extent may be gathered from the two following
instances:—During the greater part of 1912 I was stationed in
Ilorin, a province which, with the exception of one division, is
considered not unsuitable for horses. Nevertheless, during the
eight completed months of my residence, ten out of the fifteen
horses in the possession of Europeans contracted trypanosomiasis,
and six died of the disease. An experiment was also made with a
view to introducing animal transport into the province. Ten
healthy donkeys were selected for this purpose, and sent to carry
loads from Ilorin town to Agugi, a village some thirty miles to the
east. Within three months all had died of trypanosomiasis. At
Zungeru, during the year 1911, twenty-five horses were treated for
this disease in the isolation camp, and of these 40 % either died
or were so seriously affected that their owners thought it wise to
part with them at the small price a native is always willing to pay
for a sick animal. At Lokoja the disease is even more serious, and
Dr. C. F. Watson has estimated that among the horses treated for
tsetse disease there, the rate of dead and incapacitated horses is
over 73 %. There can, moreover, be little doubt that the
disease is spreading; a fact proved by the additions made from
time to time to the list of localities in which the Government declines
to compensate officers for the loss of horses which have died from
trypanosomiasis contracted in the discharge of official duties.
2
Difficult as it is to estimate the losses among horses belonging to
the small number of Europeans stationed in the Protectorate, it is
altogether impossible to ascertain those of the natives. They must,
however, be very great, and their magnitude may be gauged by the
experiences of the Emirs who attended the Coronation Durbar at
Zaria in June, 1911, one of whom is said to have lost 70 per cent,
of his horses. Before the British undertook the administration of
Nigeria, certain tracts of country known to be unsuitable for horses
were sedulously avoided. Europeans, however, are compelled to
penetrate these districts in the discharge of their duties, and are
followed by the native officials, not only at the sacrifice of their
own animals, but also greatly to the detriment of the whole
country, since the infected horses returning home, or passing
through other districts, spread the disease in every direction.
Besides horses, the cattle, sheep, goats and dogs are also affected.
In their long trek to the coast, the herds of cattle from the north
carry trypanosomiasis with them, maintaining the infection in old
localities and introducing it into new. The herdsmen are quick to
detect the symptoms of the disease, and, if they appear in one of
their animals, are accustomed to slaughter the beast wherever they
may happen to be. The fate of those that sicken is to be butchered
by the roadside, and it is a common sight to see a carcase cut up
and laid out for sale far away from any native town. Ilorin is
situated at a point on the main caravan road where the routes from
Kano and Sokoto converge, and the majority of the animals
slaughtered there for the market were found on examination to be
suffering from trypanosomiasis.
In Table I the results of the examinations of thirty-five of these
animals is shown : —
Table I.—Trypanosomes found in the blood of thirty-five sick animals being slaughtered for the
Ilorin market
Host T. brucei ]
T. vivax T. nartum or j
pecorum !
Double
infection
Cattle—Fulani breed. 1 i
18 j 1 j
1
Sheep and rams . i
8 j i I
Goats .
!
1
j 4 i -
. _
Totals . 2
1
; 30 1 2
1 «
3
In November, 1911, Sir David Bruce published in No. 31 of the
Sleeping Sickness Bulletin 1 a list of identifications of fifteen cases
collected by me in Northern Nigeria of trypanosomiasis in horses.
Since then I have added to these a number of fresh cases, both in
horses and other domestic animals, bringing the number up to
eighty-eight. An analysis of eighty-six of these cases, in which the
infection was natural, is given below (Table II).
Table II.—Showing the infecting agent in eighty-six cases of naturally acquired trypanosomiasis
in domestic animals in Northern Nigeria
Host
T. brucei
T. vivax
1 T. nantim or T. theileri
Double
p e corum
infections
Horse .
14
18
; * i ...
3
Donkey ... .
2
2
J »
Cattle—
1
'
1
1 1
Fulani breed .
.
18
1
1
1
Dwarf breed .
2
!
1
Sheep and ram .
1
8
1 i
II...
Goat .
4
1
...
Dog .
...
|
1
1 1
' 1
Totals
1 18
1
53
1
IO 1
4
As it was impossible in Northern Nigeria to carry out systematic
series of experiments on animals or to attempt cross immunity tests,
the morphological characters of the trypanosomes found in blood-
films had to be relied on for identification. By this means it was, of
course, impossible to distinguish between the more closely allied
species. T. theileri was observed once, but with this exception the
trypanosomes detected fell naturally into three main groups, and,
as individuals of each type had previously been examined by
Sir David Bruce, his identifications have been adopted. 2
T. brucei.* In twenty cases of natural infection trypanosomes
were found which closely corresponded with the type described and
figured by Sir David Bruce as T. brucei A In four cases (three
horses and one donkey) posterior nuclear forms were detected, and
• This trypanosome is probably that for which Prof. Stephens and Dr. Blacklock have
recently proposed the name T. ugandae. (Proc. Roy. Soc., B, Vol. LXXXVI, pp. 187-191).
4
in guinea-pigs and rats inoculated from three of these cases they
appeared at certain stages of the disease in relatively large numbers.
Posterior nuclear forms have been described in T. equiperdum ,
T . pecaudi and T. brucei , as well as in T. rhodesiense. Their
occurrence in these cases of trypanosomiasis in Nigeria does not
therefore assist in precise identification. Sleeping sickness, how¬
ever, is either altogether absent or quite uncommon at Zungeru and
Ilorin, where the majority of my cases were found. In a number
of films the long forms of the parasite were observed to have their
posterior extremities peculiarly blunt and almost rectangular. No
attempt was made to plot a curve to represent the variation in size
of this trypanosome. Great variations were observed from day to
day in the relative percentages of short and long forms, as is
indicated in the tables given below, where the different forms were
enumerated in the case of a horse, and in those of a rat and a
guinea-pig inoculated with his blood. (See table on page 5.)
The disease set up by these trypanosomes appears to be
extremely fatal. Of the eleven horses and two donkeys suffering
from this form of trypanosomiasis of which I have complete
records none recovered. In horses the disease seems to occur in two
forms, some cases end fatally in two to four weeks, others linger on
for as many months. All the cases in which posterior nuclear
forms were observed were of the acute type, death occurring within
a month of the onset of the disease.
The general symptoms in horses were fever, emaciation, and
more or less pronounced oedema of the legs, belly and scrotum.
The appetite was irregular, but was often ravenous almost up to
the hour of death. In the later stages restlessness was a distressing
feature. Inflammation of the eyes was observed in two cases out
of fourteen, and a greenish discharge from the nose, coughing up
of mucus, and a herpetiform eruption on the upper lip occurred
each in one case. The animals presented a dejected appearance in
the early stages of the disease, their heads drooped, they dragged
their hind legs in walking, moved slowly and with difficulty and
were apt to stumble. They were notably insensitive to pain, as
for instance, to the prick on the ear necessary to obtain blood for
examination. Finally they became so weak and wasted that they
could not rise.
5
Natural infections with trypanosomes of this type were found in
the horse, donkey, Fulani cattle, and sheep. The parasite was also
successfully transmitted by inoculation to a cow of the dwarf breed
found in pagan districts of Nigeria. (See page io.)
Date
*, 1912
Horse (No. 35)
Rat
Guinea-pig
Forms with
long, free
flagella
Stumpy and
intermediate
forms
Forms with
long, free
flagella
Stumpy and
intermediate
forms
Forms with
long, free
flagella
Stumpy and
intermediate
forms
July
8 ...
* 4 %
76%
V.
9 ...
4 *%
58%
...
10 ...
Rat injected from horse.
Guinea-pig
injected from
(Trypanosomes first
horse. (Trypanosomes
appeared
in the blood
first appeared in the
on July if
>)
blood on
July 18)
13 ...
* 9 %
7 i%
16 ...
Trypanosome
s very scanty
89%
«•%
...
17 ...
98%
2%
-
18 ...
5*%
48%
•5%
85 %
19 ...
*5%
^5 %
40%
60%
20 ...
74%
*6%
>0%
9 °%
$ 6 %
44 %
21
8%
9*%
l 6 'S%
83 - 5 %
27 ...
6 % I
94 %
88%
’ 2 %
Died
July 21
V
1
28 ...
Died
29 ...
30 ...
58 %
42 %
Aug.
1
85 %
• 5 %
4 •••]
69%
3 '%
» -1
Died
1
i
T. vivax . By far the most common form of trypanosome in
my series of cases was one presenting the morphological features of
T. vivax. It occurred in fifty-six of my eighty-six cases, and in
fifty-three cases was the only trypanosome present, being but once
associated with T. brucei and twice with T. nanum or pec or urn.
6
This parasite was found in the following hosts:—Horse, donkey,
Fulani cattle, dwarf cattle of Nigeria, sheep, goat and dog. From
the fact that it was found in the dog, it is probable that this
trypanosome is the parasite to which Ziemann 4 gave the name
T. vivax , and not the allied T. cazalboui described by Pecaud as
occurring in Dahomey in horses. For Laveran and Mesnil say:
' Le fait que Tr. Cazalboui ne peut 6tre inocul6 ni au singe, ni au
chien, ni aux petits rongeurs, permet de distinguer facilement ce
trypan, des especes qui s’en rapprochent au point de vue morpho-
logique, mais qui sent pathog&nes pour ces animaux.' 5
In horses the disease set up by this parasite seems to be mild in
Northern Nigeria. Of the fifteen cases of which I have complete
records only one died, whereas fourteen recovered. The general
symptoms were fever, emaciation and oedema of the legs, belly
and scrotum, sometimes well marked, at other times scarcely
perceptible. The animals presented a dejected appearance, moved
awkwardly and reluctantly, and appeared as though tired. In
severe cases they were somewhat insensitive to pain. The appetite
was always good. In one case cough was a symptom, and in
another nasal catarrh. No affections of the eyes were observed.
Three cases were met with in donkeys, and of these two died and
one recovered. The case of one of those that died was, however,
complicated by a concurrent infection with T. brucei. Eighteen
cases were found amongst Fulani cattle. Most of them were
animals treking down towards Lagos from Kano or Sokoto which
had sickened on the road, and were therefore about to be butchered
for the Ilorin market. Their native owners believed them to be
dying, and were therefore having them slaughtered so as to
anticipate death; but, beyond this assertion, I have no data from
which to gauge the mortality from the disease in this breed of
cattle. Both the dwarf cattle which came under my notice suffering
from T. vivax , however, recovered. Similarly the eight sheep and
rams I studied were animals about to be slaughtered so as to
anticipate death, and the same remark applies to the four goats.
The one dog in which T. vivax was found died.
T. nanum or fecorum . In a few cases small trypanosomes
9-1 2 /x in length without free flagella were found in blood-films,
and these have been identified as T. nanum or fecorum. ‘ It will be
1
remembered the name T . pecorum was introduced by Bruce to cover
the group of trypanosomes of which the T . dimorphon of Laveran
and Mesnil and T. congolense are members, and that nanum is only
distinguishable from pecorum by animal experiments/ 6
Trypanosomes of this type were met with in thirteen cases, in
ten alone and in three associated with T. vivax or T. brucei. Ten
of these cases were horses, two Fulani cows, and one a ram. Of the
ten cases in horses one had a concurrent infection with T. vivax
and another with T . brucei , leaving eight uncomplicated cases, of
which one died, three recovered, and the remaining four had
unknowm issues.
The number of cases of this variety of trypanosomiasis that
came under my notice does not, therefore, enable me to give any
general account of the symptoms of the disease in Northern
Nigeria. It may, however, be noted that oedema was present only
to a slight extent in the cases seen, although lachrymal and nasal
discharges were relatively more frequent than in infections with
either T. vivax or T. brucei.
T. theileri. T. theileri was observed once in a cow of the dwarf
breed found in Nigeria. The animal was feverish for a few days
and wasted considerably, but soon recovered from the infection.
The period when trypanosomiasis is most common is undoubtedly
the rainy season, which corresponds to the time during which the
tsetse flies, which in the dry season retire to the banks of the streams
and rivers, are most widely distributed over the country. The
accompanying chart of the rainfall and the number of admissions
of horses to the isolation camp at Zungeru for the year 1911
illustrates this point, although the number of cases dealt with is too
small to form a reliable index. A better proof is afforded by the
fact that the natives congratulate themselves at the end of the
rainy season that those of their horses which have withstood the
disease so far will be safe until the rains recommence. The dry
season is also the time of year selected by the herdsmen of the north
to undertake their long trek south with their cattle and sheep.
The province of Ilorin in Northern Nigeria is peculiar, inasmuch
as, whilst G . palpalis and G. tachinoides are distributed all over it,
tsetse flies of the morsitans group— G. submorsitans and G. longi -
palpis —are restricted to one, the Patigi or eastern, division. In
8
my series of cases of trypanosomiasis collected at Ilorin, twenty-
four were animals that had never been into the Patigi division, and
of these fourteen were infected with T. vivax , six with T. brucet
and four with T. nanum or pecorum. The Patigi division is
shunned by all herdsmen, and it is generally recognised that horses
can neither live there nor be taken into the district without
contracting tsetse disease. Two horses that had lived for years in
Ilorin town were taken to Patigi during my last tour of service.
Chart showing the rainfall in inches (continuous line) and the number of cases of
trypanosomiasis of horses admitted to the isolation camp (dotted line) at Zungeru
during 1911.
Both returned infected with T . brucet , and both died shortly after
wards. In this connexion it should be mentioned that Bruce
concludes that, in Uganda, ‘ The carrier of Trypanosoma vivax is
probably Glossina palpalis and that G. morsitans is known to
transmit T. brucei. It is possible that in the native towns and
European stations flies of the genus Stomoxys , which abound in
these places, may play a part in the transmission of trypano¬
somiasis. The horse (No. 42), for instance, had not been within
9
two miles of any spot known to be haunted by tsetse flies for five
weeks previous to the onset of his symptoms, and it is practically
certain that during this period he could not have been bitten by
tsetse flies. He was, however, tormented by Stomoxys flies, which
were exceedingly common at the time. Both 5 . nigra and
5 . calciirans have been taken at Ilorin.
Treatment. The treatment usually adopted in Northern Nigeria
for all cases of trypanosomiasis in horses is the administration of
arsenious acid and perchloride of mercury by the mouth. This is
the routine treatment adopted at the isolation camps at Zungeru
and Lokoja. The doses usually employed are nine grains of the
arsenic and three ounces of a i in i ,ooo solution of the perchloride
of mercury daily, divided into three doses; but I have also tried
much larger doses, administered for a few days at a time only, in
the hope that the sudden shock of arsenic might eliminate the
trypanosomes before they had time to react to the drug, and with a
view to avoiding the cumulative action on the host that has certainly
sometimes proved fatal. It is doubtful whether this treatment has
any beneficial results. In my experience it has not saved a single
animal infected with T. brucei, and it is impossible to judge of its
action on T. vivax, as this form of trypanosomiasis is as a rule not
fatal, the animals recovering even when untreated. At one time
it was asserted that such treatment accounted for the cure of 50 %
of the cases at Zungeru. This, however, was before it was
known that half the trypanosomiasis there was due to T. vivax , and
was capable of spontaneous cure. The higher mortality recorded
at Lokoja is probably accounted for by the larger percentage of
cases of trypanosomiasis due to T. brucei which occur at that
station.
Intravenous injections of antimony have met with no greater
success in trypanosomiasis due to T. brucei. Organic arsenical
compounds have been used in only a few cases, the high cost of the
injections and the technical difficulties of administration standing
in the way of the general use of these preparations until it is
certain, not only that a cure will result, but also that immunity
against future attacks will be established.
Arsenic and mercury have been used as prophylactics also, but
without success.
to
No immunity follows an attack of trypanosomiasis, reinfections
with the same or a different species of trypanosome being met with.
The dwarf cow (No. 48) in my series was infected successively with
T. theileri , T. vivax and T. brucet , and finally died of the last form
of trypanosomiasis. The donkey (No. 47), after recovering from
an infection with T. vivax , was infected by injection with
T. brucet , and succumbed to the disease.
Treatment of any sort is, however, applicable only to Govern¬
ment stock and horses in the possession of Europeans. The great
majority of cases, occurring in animals which are the property of
natives, cannot be brought under treatment. It might be possible,
nevertheless, to limit the spread of the disease by the establishment
of isolation camps, and much might be effected by systematic
clearing along the roads. The main caravan routes especially
should be cleared, and inducement offered to the natives to farm
the land on either side of the roads.
Dwarf cattle . In certain districts in which the Fulani cattle do
not live a few dwarf cattle are found, which, because they occur in
pagan country, are often referred to as ‘ pagan cattle. 1 In Ilorin
they are kept in all the districts occupying the south-eastern corner
of the province, from Ofa Ora to Ejiba on the north, and
extending west along the southern boundary as far as Oke Awra.
Dr. Foy, who made some interesting observations and experiments
on a bull of this breed, thus described his physical characters.
1 The body frame/ he says, 4 was thick-set and broad and compara¬
tively long for its height, the legs thick and short, the neck thick
and short, the head short, and the horns short/ 8 The accom¬
panying photograph of a bull and cow brought from Awtun will
help to give an accurate idea of the appearance of these singular
animals (PL III, fig. 5).
The most remarkable point about these dwarf cattle, which are
bred for slaughter only and are not used for milking, is that they
live in districts in which the Fulani cattle cannot exist. For this
reason it has been conjectured that they have acquired a high degree
of immunity to trypanosome infections. Dr. Foy, who, as already
stated, experimented on a bull of this breed, concluded: —
4 (1) That a certain breed of cattle found in pagan districts possess a high degree of natural
immunity in that they may harbour the trypanosome in the blood and yet keep in good condition and
II
show no signs of the disease, nor do they die from the infection so long as their environments are
favourable. These environments are a free life, with ample food, especially plenty of gTeen grass.
Confinement, poor feeding, and hard exercise, tend to make the disease manifest itself clinically.
‘ (2) That such domesticated cattle may act as a reservoir of infection since the blood may
prove infective at such times when clinical symptoms manifest themselves, although the trypanosome
may not be found on making a microscopical examination.
4 (3) That the subcutaneous injection of 26 c.c. of serum obtained from the blood of one of
these cattle when manifesting no clinical symptoms of the disease did not prove infective when
inoculated into a calf, nor did it act as a prophylactic when the calf was exposed to natural infection
subsequently/
He adds, in another part of the same paper, ‘ all strains of trypano¬
somes used or met with in the work were of the T. brucet type.’
At Ilorin I was able to carry out only two experiments with these
cattle, but as the subject requires careful investigation, and promises
to provide results of practical importance, I should, perhaps, place
them on record here. On May i ith, 1912, two of these dwarf cattle
(a cow and a bull) arrived at Ilorin from Awtun. As they had
treked up all the way by road, they arrived in rather poor condition,
but seemed, on the whole, to have stood the unusual exertion
wonderfully well. On May 13th I made a thorough examination
of the blood of each without detecting any trypanosomes, and four
days later injected a rat and a guinea-pig each with 1 c.c. of blood
taken from the cow. Neither of these animals developed any
symptoms of disease, and, although they were kept under close
and constant observation for four weeks, trypanosomes were never
seen in their blood. The subsequent history of the cattle is given
below: —
Dwarf Bull (No. 50)
May nth, 1912.—Arrived at Ilorin.
May 13th, 1912.—Blood examination—negative.
August 30th, 1912.—Trypanosomes (T. vivax) in the blood. Bull looks well. Untreated.
September 15th, 1912.—Blood examination—negative. Bull, however, looks ill.
September 19th, 1912.—Bull died of anthrax. No trypanosomes found in the blood nor in smears
made from the organs after death.
Dwarf Cow (No. 48)
May nth, 1912.—Arrived at Ilorin.
May 13th, 1912.—Blood examination—negative.
12
May 17th, 1912.—Blood examination—negative. Subcutaneoui injections of the cow’s blood
made into a rat and a guinea-pig.
Rat under observation twenty-eight days, but never showed trypanosomes.
Guinea-pig under observation twenty-eight days but never showed trypanosomes.
August 30th, 1912.—Cow feverish and emaciated, no oedemas. Trypanosomes (T. tbeiUrt and
T. vivax) present in the blood. Untreated.
September 7th, 1912.—Trypanosomes still present in the blood. Subcutaneous injection of two c.c
of the cow’s blood into a guinea-pig.
Guinea-pig under observation nineteen days. Accidentally killed. Never showed
trypanosomes.
September 18th, 1912.—Blood examination—negative. One c.c. of the cow’s blood injected
subcutaneously into a rat.
Rat under observation twenty-two days but never showed trypanosomes.
September 27th, 1912.—Cow looks well, and her blood has been free from trypanosomes since
September 15th. Two c.c. of blood from donkey (No. 45), heavily infected with T. brucei ,
injected into cow subcutaneously.
October 8th, 1912.—Trypanosomes numerous (T. brucei). The blood was negative up to October 5th.
November 3rd, 1912.—Cow died. Trypanosomes still present in the blood but scanty.
Both animals developed a natural infection with T. vivax , and
apparently recovered, for they ceased to show trypanosomes in films
made from the peripheral blood. The cow in addition had a
concurrent infection with T. theileri. The cow was then successfully
infected by injection of blood from a donkey heavily infected with
T. brucei , and, although the parasites soon became scarce in her
blood, she died five weeks after the injection still showing a few
trypanosomes in the peripheral blood.
It would be rash to draw definite conclusions from such experi¬
ments, but so far as they go they do tend to show that this breed
of cattle, like horses, possesses an immunity to T . vivax , inasmuch
as although these parasites may appear in their blood they do not
produce a rapidly fatal disease. The immunity does not appear,
however, to extend to T. brucei. The question then arises how do
these cattle live in districts where the Fulani herdsmen dare not
take their cattle to graze? The dwarf cattle are generally to be
found in the immediate vicinity of villages, and unlike the Fulani
cattle do not wander widely over the country in search of pastures.
It was thought possible, therefore, that they might thus escape the
attacks of G. submorsitans , the species of tsetse fly whose distribu-
tion (as already pointed out) is coextensive with the areas habitually
shunned by the Fulani herdsmen. In consequence, collections of
flies were obtained at Odo Okeri, Eri, Oro and Oke Onio, from
spots where the cattle were actually grazing. In none of these
collections was G. submorsitans present, although both G . palpalis
and G. tachinoides were taken. I have to thank Mr. T. A. G.
Budgen for very kindly superintending the work of a collector who
was sent with him for this purpose. I can only hope that at some
future date, and under more favourable conditions, it may be
possible to make a thorough investigation of this interesting subject.
In conclusion, I would like to express my thanks to Mr. E. C.
Duff for affording me an opportunity of examining the dwarf breed
of cattle; to Dr. C. F. Watson and Dr. W. Morrison for very
kindly sending me blood-films from Lokoja; to Dr. G. R. Twomey,
who relieved me at Ilorin, for following the last stages of my
experiments; and to Serjt. Moore for the careful record kept by
him of the cases of trypanosomiasis occurring at Zungeru.
A tabular synopsis of the cases of trypanosomiasis in domestic
animals, collected in Northern Nigeria, is appended.
*4
Identifications by Sir David Bruce.
i6
Sept., 1912 T. vivax and Died Ill about 2 month?.
T. brncei
Synopsis or Casf.s or Trypanosomiasis of Domestic Animals. Collected in Northern Nigeria, continued .
'7
|8
Synopsis ok Cases of Trypanosomiasis ok Domestic Animals. Collected in Northern Nigeria.— continued .
19
20
SUMMARY
1. Trypanosomes presenting the morphological characters of
T. brucei y T. vivax , T. nanum or pecorum y and T. theUeri have
been found in Northern Nigeria in the blood of domestic animals;
T. brucei in the horse, donkey, Fulani cattle, dwarf cattle and
sheep; T. vivax in the horse, donkey, Fulani cattle, dwarf cattle,
sheep, goat and dog; T. nanum or pecorum in the horse, Fulani
cattle and sheep; and T. theileri in the dwarf cattle.
2. T. vivax is the most common form at any rate in the south¬
western portions of the Protectorate, being present in fifty-five out
of eighty-four cases collected in Ilorin province, the Niger province,
and at Lokoja.
3. Of twenty Fulani cattle treking down towards the coast
from the provinces of the north, and found to be suffering from
trypanosomiasis, eighteen harboured T. vivax.
4. In horses T. vivax produces a much less serious disease than
T. brucei. Of fifteen cases infected with T. vivaxy of which the
records are complete, fourteen recovered. Of eleven cases infected
with T. brucei none recovered.
5. The dwarf breed of cattle found in certain tsetse-haunted
districts of Northern Nigeria, and credited with a natural immunity
to trypanosomiasis, while apparently recovering from infections
with T. vivaxy succumb to T. brucei.
REFERENCES
1. Sleeping Sickness Bureau Bulletin, Vol. 3, No. 31, p. 422, 1911.
2. Ibid.
3. Reoorts of the Sleeping Sickness Commission of the Royal Society, No. XI.
4. H. Zhmann. Centralbl. f. Bakter., 1, Orig., t. XXXVIII, 1905.
5. A. Lavkkan et F. Misnil, Trypanosomes et Trypanosomiases, p. 552, 1912.
6. Sleeping Sickness Bureau Bulletin, Vol. 3, No. 31, p. 423, 1911.
7. Reports of the Sleeping Sickness Commission of the Royal Society, No. XI.
8. 4 A third report on experimental work on animal trypanosomiasis/ by H. Andrew Foy,
D.P.H., in The Journal of Tropical Medicine and Hygiene, October 16th, 1911.
22
EXPLANATION OF PLATE I
Fig. i. Horse (No. 35). Trypanosomiasis ( T. brucei).
July, 1912.
Fig. 2. Horse (No. 42). Trypanosomiasis ( T. brucei ).
October, 1912.
Ilorin,
Ilorin,
Fig- 3
Fig. 4
*4
EXPLANATION OF PLATE II
Donkey (No. 44). Trypanosomiasis ( 7 \ brucei ). Ilorin,
August, 1912.
Donkey (No. 45). Trypanosomiasis ( T . brucei ). Ilorin,
September, 1912.
Annals Trap. Msd. & Parasite?Vol. Vll
26
EXPLANATION OF PLATE III
Fig. 5. Dwarf cattle. Ilorin, May, 1912.
Annals Trop. Med. & ParasitolJ'ol. VII
PLATE III
‘7
FURTHER MEASUREMENTS OF
TRYPANOSOMA RHODESIENSE
AND T. GAMBIENSE
BY
J. W. W. STEPHENS, M.D. Cantab., D.P.H.
AND
H. B. FANTHAM, D.Sc. Lond., B.A. Cantab.
(Received for publication 21 December , 1912)
In our former paper (May, 1912) on this subject, we came to the
conclusion that it was advisable in measuring trypanosomes to
confine our observations to those from a single animal, for example,
a rat, as although it could not be definitely proved that the size of
the trypanosome varied in different animals, yet it appeared likely
from the general consensus of opinion that this might be so. From
the statistical side the criticism has also been brought that samples
of twenty at any particular time are too small. In our present series
of measurements therefore, we have, as far as possible, met these
objections by measuring always from a single animal, a rat, and
by measuring one hundred trypanosomes each day for the first ten
days of the infection. We may here briefly repeat our method, as
it has been subjected to some criticism.
1. We project the trypanosomes on a screen in a dark room
and trace them, instead of drawing them with a camera lucida.
It has been objected that this method cannot be used in the wilds
of Africa, but we never stated that it could or should, and that is no
reason why we should not use it in a laboratory. Our critics might
as well object to our using electric light.
2. We measure the trypanosomes by means of the * tangent
line * method. We believe that as this method is the most accurate
known—our critics have not attempted to deny this—we are again
amply justified in preferring it to the less accurate compass method,
even though the difference may be only 1 or 2/1. It seems to us
hardly a matter for argument that if it is worth measuring 1,000
trypanosomes at all, it is worth doing so accurately, especially if
length is to be considered a criterion of specificity.
We believe that it is important to give the actual data as to
measurements as Bruce does, and not simply averages, as the actual
data are necessary for a closer analysis than averages permit. We
give first, then, the fundamental data for each trypanosome
( T . rhodesiense and T. gambiense) and the tables compiled from
them. We shall subsequently make a comparative analysis of the
two sets of figures.
Tabli I.—Distribution in respect to length of 1,000 non-dividing individuals of Trypanosoma rboJesieitse from a single white rat.
foil Day ... id
These numbers refer to the consecutive groups of twenty trypanosomes, summarised on each line.
30
Table II.—Summary of measurements (in microns) of lengths of 1,000 individuals of
Trypanosoma rhodesiense from a single white rat.
Maximum
Minimum
Averages
of each 100
Averages
of each 20
Range of
averages
of each 20
I
30
21
26-0
2
28
*9
24-05
I st Day 3
30
*9
25-14
25-6
1 '95
4
29
18
24-1
5
3 *
22
25-95
6
29
20
25-65
7
29
23
26-1
2nd Day 8
28
16
22-98
21*1
5-25
9
3 °
16
20-85
IO
26
16
21-2
ii
27
19
23-25
12
27
14
22-2
3rd Day 13
3 °
18
22-51
22-85
1-25
H
26
*9
22*0
*5
29
16
22-25
16
29
18
23*0
*7
26
18
22-05
4th Day 18
26
16
22-16
20-95
2-65
*9
26
17
21*2
20
28
*9
23-6
21
3 °
16
22-8
22
28
21
2 5 -2
5th Day 23
3 i
*9
24-72
25*35
2*7
24
28
17
24*75
2 5
34
20
25*5
26
25
l 7
22-0
27
27
'7
23-1
6th Day 28
29
18
24-19
24-45
3-85
29
30
21
25-85
3 °
29
21
25-55
3 *
22
16
19-25
32
24
17
19-9
7th day 33
29
18
2I-7I
23-05
3*95
34
28
20
23-2
35
28
l 7
23*15
i
3 6
32
15
25-1
37
3 °
14
24-8
8th Day 38
33
18
26-I5
27*75
2-95
39
33
20
26*2
40
32
20
I
26-9
41
29
17
25*45
42
3 i
*9
25-85
9th Day 43
32
16
*5-67
25*5
o *45
44
30
22
25*9
45
29
>5
25-65
46
3 i
18
25-9
47
29
20
* 5-5
10th Day 48
3 i
24
25-98
26-95
i *45
49
30
19
* 5-45
50
33
17
26*1
Range =27-75—
19-25=8-5
3 1
Table III.— 7 . rbodesUnse, in which the trypanosomes are arranged in Bruce’s three
groups (0) 13-21 n ; ( b ) 22-24 fi ; ( c ) 25/A and upwards.
Day
1
2 '
3
4
| 5
6
i
1 7
8
1
9
■o
Totals
Stumpy
I 3 - 2 IM
12
41
37
45 1
.7
16
53
x 5
9
8
253
Intermediate
22-24 M
i
2 5
*9
40 !
28 |
3 1
4 °
29
18
16
21
267
Long
25-36/*
63
40
23
2 7 j
1
| 52
44
1 18
i
67
75
D
480
100
100
| 100
100
100
100
100
i 1
100
1
100
IOO
1,000
This table shows very clearly what we have already pointed out,
namely, the great variation in the figures for each group on
particular days. Thus on the tenth day there were 8 % of stumpy
forms, while on the seventh day there were 53 %. This seems to
us to make it perfectly obvious that when a sample is taken at
random from an animal on any day an erratic factor is introduced.
We proceed to represent the preceding results graphically.
Chart I shows a curve of measurement of Trypanosoma
rhodesiense. The distribution, by percentages in respect to length,
of the 1,000 non-dividing specimens of the trypanosome is plotted.
The parasites were taken from the peripheral blood of a rat. One
hundred trypanosomes were measured each day for ten consecutive
days of infection (vide Table I).
Chart I. —Graphical representation of the distribution of the lengths of 1,000 7 . rbodtiiens *
from one rat.
We next proceed to give corresponding tables for T. gambiense.
Table IV.—Distribution in respect to length of i ooo non-dividing individuals of Trypanosoma oambitnse from a single white rat.
2 Z *0
Totals
34
Tablx V.—Summary of measurements (in microns) of lengths of 1,000 individuals of 7 .
gambitiut from a single white rat
Maximum
Minimum
Averages
of each 100
Averages
of each 20
Range of
averages of
each 20
I
2 7
*7
wap
2
3 2
21
ist Day 3
28
18
24*16
■
3-85
4
2 9
22
5
29
21
mmSmm
6
3 2
22
267
7
ji
x 7
26-35
2nd Day 8
30
24
26*65
26-95
0*85
9
30
22
26*2
IO
30
2 3
27-05
I!
2 9
18
24*3
12
3 2
l 7
24-65
3rd Day ij
28
16
24*14
22*0
4*35
H
28
*7
2 3*4
*5
.. 30
21
26-35
16
36
21
2675
*7
3 i
21
27*25
4th Day 18
3°
! 9
26*42
2 5‘35
j * 9
19
3 2
21
2 7'*5
20
3 2
21
25*6
21
26
*9
22* 15
22
2 9
16
24*4
5th Day 23
2 9
20
23-87
24*75
2*6
*4
3 1
*9
24*55
2 5
30
18
2 3*5
26
3 2
20
24*4
2 7
26
*7
22*85
6th Day 28
28
«9
22*88
2 3*35
2*6
29
28
*7
22*0
30
2 5
*9
21*8
3*
2 5
16
3 2
2 4
18
7 th Day 33
26
*7
21*07
*’4
34
2 7
16
35
2 7
16
36
3°
20
26*0
37
3i
20
25*8
8th Day 38
3 1
22
26-37
27*0
2*25
39
3 2
21
27-65
40
2 9
20
25*4
4 *
2 9
21
257
42
3o
*9
26*25
9th Day 43
3 2
21
26*17
25-65
1*65
44
30
18
2 5*95
45
33
22
2 7*3
46
3 2
>9
27*5
47
34
21
2 7*7
10th Day 48
3 2
l 7
26-94
26-55
1*7
49
3 2
17
26*00
5°
33
21
26-95
Range **277—
2 °* 2 5 - 7*45
35
Table VI.— 7 . gambieme , in which the trypanosomes are arranged in Bruce’s three groups.
Day
I
fl
3
B
5
B
,0
Totals
Stumpy
13-11/*
l 7
1
25
10
23
29
57
6
5
182
Intermediate
21-24 M
1
40
16
2 9
49
33
*5
20
■
1
12
1
270
Long
* 5 - 3 « P
43
83
79
100
IOO
IOO
IOO
IOO
!
IOO
1
IOO
IOO
IOO j
IOO
1,000
Here again we note a great variation in the figures for each
group, for example, on the second day I % of stumpy forms, and
on the seventh day 57 %. The differences are due, according to
some authors, to a cycle in the vertebrate host.
In Chart II we give a curve representing the distribution, by
percentages in respect to length, of the 1,000 non-dividing
specimens of Trypanosoma gambiense, taken from a rat. One
hundred trypanosomes were measured each day for ten consecutive
days of infection (vide Table IV).
Chabt II.— Graphical representation of the distribution of the lengths of 1,000 7 . gambiense
from one rat.
3 6
Comparing now T. gambiense and T. rhodesiense we get the
following tables.
Table VII
Average length
Maximum
Minimum
T. gambiense
14-87/*
3 6 *°M
16*0/4
7 . rhodesiense
24 - 12 /i
34 '°M
I4*o/a
Table VIII.
—Comparison of distribution of the trypanosomes according to Bruce’s groups.
13—2IM
22—2^
25ft and upwards
T. gambiense
■ 8-1%
27-0 %
54 - 8 %
T. rhodesiense
* 5 ' 3 %
267 %
48*° %
Table IX.—Distribution by Octiles of both 7 . gambiense and T. rhodesiense.
125th
250th
375 th
500th
625th
75 oth j
875th
T. gambiense .
2I/i
22/4
2 5 M
26/4
29/4
T. rhodesiense .
20/1
21/4
24/4
26/4
27/4
28/4
DISCUSSION OF RESULTS
The fact that we are dealing with a dimorphic trypanosome,
the dimorphic nature of which is not thoroughly understood, is no
doubt responsible for the difference of opinion as to procedure and
for the different results obtained. Such difficulties probably do
not arise in the case of a monomorphic trypanosome.
I. THE SIZE OF THE SAMPLE
If Tables II and V be examined, it will be found that on one day
the average values of five samples of twenty trypanosomes may
vary by as much as 4‘35/* in the case of T. gambiense and by
5*25/1 in the case of T. rhodesiense. Also the differences between
37
the average value for a sample of ioo on any particular day and
any of the samples of 20 for that day may vary by as much as
2*21/4 (third day) in the case of T. gambiense , and 3*12/1
(second day) in the case of T. rhodesiense , but in the majority of
cases it is only about 1 /*. From this it may be inferred that a
sample of 100 is a fairly reliable one when trypanosomes are
plentiful in the blood; it will, however, not be so good a sample as
when trypanosomes are scanty.
II. THE INFLUENCE OF THE DAY OF INFECTION
As was indicated clearly in our previous paper, the day of
infection in an acute trypanosomiasis (when death ensues in about
ten days), such as that with which we have been dealing, is very
important in determining whether the trypanosomes are short or
long. Thus on examining our results (arranged according to Bruce's
groups) in Table VI, we find that stumpy forms vary from 1 % on
the second day to 57% on the seventh day, and this variation is,
of course, reflected in the average values of 100 for those days,
viz., 26*65/4 and 21*07/1 respectively. Whether these differences
occur in so marked a degree in a chronic infection we are not in a
position to state.
III. THE INFLUENCE OF THE ANIMAL HOST
As the day is of prime importance, it is impossible to say
whether the length of any trypanosome varies markedly in different
hosts. So far as we can see, this could only be determined by
measuring 1,000 trypanosomes (if this number suffice) from each of
the hosts in question. Consequently we consider it is advisable at
present to measure from the same species of animal if comparisons
are to be of value.
IV. COMPARISON OF OUR RESULTS WITH THOSE OF
OTHER OBSERVERS
Curves for T. rhodesiense have been constructed by Bruce and
his collaborators (1912), by Kinghorn and Yorke (1912), and by
ourselves. There is a fair correspondence between those of Bruce
and ourselves, but none between those of Kinghorn and Yorke and
3 »
ourselves. We cannot, unfortunately, in the present state of our
knowledge, explain these differences satisfactorily. We believe
that they must be due to difference of method, namely, that other
observers have taken different species of animals on a variety of
days.
We ourselves have now completed three curves (i) based on 1,000
trypanosomes from various animals, but including 600 from rats;
(2) based on 600 trypanosomes from rats alone, where samples of
twenty were taken on a variety of days from several rats; and now
finally (3) 1,000 trypanosomes comprising /oo a day from one rat
for ten days. These three curves have this in common, that each
one has its main peak at 26/4. Further, the agreement is most close
between curves (2) and (3) based on rats alone.
We have given previously our reasons for believing that the third
method is the best, and the agreement that exists between our
curves indicates, we believe, the consistency of our method.
We must admit that Kinghorn and Yorke’s three curves are also
consistent, but it is noticeable that in their measurements, though
different animals were used, the total number of trypanosomes from
each animal was the same in each case. There is further the
possibility that trypanosomes direct from the natural vertebrate
hosts or the fly have a different character from those that have been
maintained in laboratory animals.
CONCLUSION
We must admit that we had hoped to be able to distinguish
between the two species, T. rhodesiense and T. gambiense , by
measuring one thousand specimens of each organism. Though these
biometric results are not sufficiently conclusive, we think that it is
generally admitted that the two species are distinct.
Microscopically, the two trypanosomes are indistinguishable
except by the posterior nuclear character of T . rhodesiense . We
believe that a curve only expresses graphically what the eye can
appreciate under the microscope, and that if two trypanosomes
cannot be distinguished microscopically, we shall not be able to do
so by measuring them. However, provided that further experience
enables observers to agree as to the best procedure, it is no doubt
59
a great advantage to have a correct graphical expression for what
is otherwise only an impression, although it may be a quite accurate
one. Further, these measurements should not be regarded as
useless, as they will undoubtedly form the basis (provided all the
protocols are given) for a critical statistical investigation in the
future.
We have pleasure in thanking Mr. Walter Stott, Honorary
Statistician to the Liverpool School of Tropical Medicine, for kindly
examining our figures.
REFERENCES
Bruck, Sir D., Harvey, D., Hamkiton, A. E., Davey, J. B., and Lady Bruce (1912). * The
Morphology of the Trypanosome Causing Disease in Man in Nyasaland.* Proc. Roy.
Soc., B, Vol. LXXXV, pp. 423-433, 2 plates.
Kinghorn, A. and Yorke, W. (1912). ‘A further Report on the Transmission of Human
Trypanosomes by Glossina morsitans , Westw.* Annals Trop. Med. and Parasitol.,
VoL VI, pp. 269-285.
Stephens, J. W. W., and Fantham, H. B. (1912). * The measurement of Trypanosoma rbodesi-
ense.' Proc. Roy. Soc., B, Vol. LXXXV, pp. 223-234. 1 plate.
4 '
A NEW MOSQUITO-PROOF AND STORM¬
PROOF HOUSE FOR THE TROPICS
BY
T. F. G. MAYER,
WIST AFRICAN MIDICAL STAFF
( Received, for publication 16 January , 1913)
Plate IV
There are all kinds of houses in the tropics. There is the old
house built on enormous piles so that its occupants may be well
away from the damp of the ground; there is the house built on
the ground; and there is the house that is raised on short pillars
only three or four feet high.
Provision for ventilation is equally varied. In olden days the
houses were designed in England, and the window area had the
same relation to the floor area as obtains here; then the window
area was increased to one-tenth that of the floor area; this proportion
was again gradually increased, until now in some places a window
area equal to one-half of the floor area is regarded as the minimum
necessary for comfort.
The discovery of the transmission of malarial fever by
mosquitos was followed by the grafting on of mosquito-proof wire
gauze to the windows and doors of existing houses, with the result
that in those in which the ventilation space was small, the rooms
were made insufferably hot and stuffy; consequently, the mosquito-
proofed window was made to open and shut and the house was
turned into a mosquito trap.
With the closing in of the house by mosquito-proofing came the
growth of the verandah and the diminution in size of the rooms,
the verandah now being used as a living room except during cold
or stormy weather.
From these few remarks it is clear that, even taking into
account the demands of local conditions, house construction in many
parts of the tropics is not based on such fixed principles to guide
architects as obtain, for instance, in England.
42
The importance of efficient mosquito protection is well known.
Its value as compared with quinine prophylaxis against malaria
may be gauged from the following figures of Celli: —
Treatment
Infected with Malaria
Mosquito protection plus quinine.
> 75 %
Mosquito protection alone .
2*5 %
Quinine alone.
20*0 %
No protection.
33*o %
The difficulty of completely protecting a house raised on pillars
from mosquitos may also be noted. Faults in the flooring nearly
always exist which allow of the entry of mosquitos.
Dr. Leonard Hill states that to render a room uncomfortably
hot and stuffy, the air in it must be hot, damp and still. In many
parts of the tropics the first two of these conditions are almost
constant, the last only therefore concerns us here.
Three factors tend to keep still the air enclosed in a mosquito-
proofed house.
1. The mosquito proofing itself, by shutting off more than
30 % of the space to which it is applied.
2. The loss of air movement owing to friction with the wires
and to the interference with the air particles which impinge on the
netting.
3. The division of the house by walls into rooms and passages
hindering through ventilation.
Factors 1 and 2 should strictly be considered together, as a
certain amount of the air impinging on the wires will ultimately
pass through the netting. But without attempting the general
consideration of the motion of the air, it is clear that for a given
uncovered area the loss of movement will be greater for a small
mesh than for a large one, on account of the greater length of edge
of wire. If the uncovered area be equal to an open square of side
a and consist of n small squares each way, then, as each side of
a wire has its effect on the air movement, the total length of edge
is X 4 ( 4 ) = 4 an.
+3
Messrs. Humphreys, Ltd., of Knightsbridge—the makers of the
first mosquito-proofed house used in Sir Patrick Manson’s classical
experiment—have designed for me a house for use in the tropics
embodying certain principles which I believe are new.
The house here illustrated (Plate IV) is constructed almost
entirely of steel; wood-work is reduced to a minimum, and, where
its use is unavoidable, is specially treated to withstand the attacks
of insects, especially white ants.
The house is built on a plinth of concrete and has a floor and a
low wall of similar material. Their surfaces are cement-rendered.
The walls are continuous with a steel framing, filled in completely
with mosquito netting made of a specially woven composite material
proved to be stronger and more durable than any other. This netting
is sandwiched between perforated metal sheets, which prevent
bulging and render entry impossible except by the spring doors at
each end of the lobby-entrances.
The roof is covered with non-conducting fibro-cement slates
and is ventilated by means of louvre windows at each end. It is
completely shut off from the room below by an asbestos ceiling.
There are no walls or partitions within the mosquito proofing,
so that whatever breeze there may be, blows straight through the
house from one side to the other. Should this wind be too cold
or too strong, as for instance during storms, it can be cut off at
will, by shutters which are provided all round the house and which
are capable of closing the whole of it. These are manipulated
from within by turning a handle, and can be made either to come
down from above, as illustrated, or to move upwards from below.
The division of the interior of the house is left to the occupant,
the ideal being the minimum required for privacy, and it is
suggested that by the use of sun blinds, screens and curtains, the
open character of the house may be preserved, but permanent brick
or stone walls may be built if desired.
It is possible to make the house of any number of stories, and
to build it on piles if desired.
In the house we have been considering that the terms wall area
and available ventilation space are almost synonymous; it must not
be forgotten, however, that the proportion between wall space and
cubic content vary greatly according to the size and shape of the
building.
+4
In buildings of the same shape the cubic content varies as
the square of the wall area, provided that the height is constant;
while a square house has a greater cubic content for a given wall
area than an oblong one.
In conclusion I have to thank Sir Ronald Ross and Professor
Simpson for their interest, and for many useful criticisms and
suggestions. I have also to thank Messrs. Humphreys, Ltd., not
only for the amount of work they have done, but also for their
courtesy.
Parasitol ., VoL III
NEW MOSQUITO-PROOF AND STORM-PROOF HOUSE FOR THE TROPICS
45
CONTRIBUTIONS TO THE STUDY OF
COLOUR MARKING AND OTHER VARI¬
ABLE CHARACTERS OF ANOPHELINAE
WITH SPECIAL REFERENCE TO THE
SYSTEMATIC AND PHYLOGENETIC
GROUPING OF SPECIES
BY
Major S. R. CHRISTOPHERS, I.M.S.
(Received for publication 30 January , 1913)
Plates V-VIII
contents
I. Variation in Colour Marking or Anopbelinae .
II. Ornamentation of the Wings in Anopbelinae .
Nodal Leucogenetic Centres of the Wing.
Intemodal Leucogenetic Centres of the Wing .
Variation in Colour Marking shown by Individuals of the same Species...
Variation in Structure other than that connected with Scales .
Structural Variation in the Immature Stages .
Variation as Displayed by Scales .
Classification, Phylogeny and Geographical Distribution of the Anopbelinae
III.
IV.
V.
VI.
VII.
VIII.
PAGE
45
5 *
56
61
65
65
72
75
81
General Conclusions
Tabular Summary
References
Explanation of Plates
85
87
93
94
I. VARIATION IN COLOUR MARKING OF ANOPHELINAE
Nature of colour marking . With few exceptions, the markings
of Anophelines are due to the varied distribution of light and dark
scales. A mixture of light and dark scales may occur, e.g., on the
wing veins of certain species or on the legs when these show
mottling; but the most striking feature of the ornamentation of the
sub-family is the occurrence of patches of either dark or light scales
forming dark or light spots respectively. Where the dark scales
predominate the appearance is that of light spots on a dark
surface; where, on the other hand, the extent of light scaling is
greater, an appearance of dark spots on a light ground is produced.
4 6
Leucogenetic centres and residual pigment areas. In a great
many species the scheme of coloration is similar, the actual markings
being brought about merely by different degrees of development of
the scheme. The process of advancing ornamentation is one of
increased whitening, and consists in the progressive appearance and
spread of pale areas which have their origin at certain fixed points
(leucogenetic centres).
Although the pale areas developing from these centres may
spread until they coalesce with neighbouring pale areas, they do
not, as a rule, do this, and there is a distinct hesitation shown at
the final extinction of intervening dark areas. Also extension
of the pale areas does not necessarily encroach equally on both
sides of a dark area, and the behaviour of markings in this respect
suggests that there exist something very like actual pigment centres
having a definitely fixed position. Points at which the pigment
seems thus to make a last stand will be referred to as residual
pigment areas, or simply as pigment centres. In some cases
centres become quite obliterated, some being apparently less
resistant than others, but even in a high degree of whitening, such
as frequently occurs in the case of the wing, where many of the
pigment areas are displayed as mere points, the number actually
present is not much reduced. A row of these pigment centres lying
near the ends of the longitudinal veins of the wing constitute the
marginal spots of Donitz, and others have an equally fixed and
characteristic nature.
By referring markings to the leucogenetic centres which produce
them (and so far as one can see, these centres of whitening are a
very real phenomenon), and to the position of the pigment centres,
the study of colour markings is greatly simplified.
Position of colour markings. The most important colour
markings are those of the palps, legs and wings. There are, of
course, variations in the coloration of other parts of the body
which may assume importance in particular species.
Almost all Anophelines have a head covered with dark scales
behind, and with a ‘ forehead ’ patch of pale white or creamy scales
prolonged forward as a tuft between the eyes. The absence of
this marking would be an important feature in any species.
In the case of the proboscis, only a few species show any
ornamentation other than the light tip due to the fact that the
47
labellae are not dark scaled like the rest of the proboscis. But a
very striking variation is seen in a few species where the outer
portion of the proboscis is pale or snowy white.
Markings of the palps. A certain number of species show palps
without any markings (unbanded palps). In some cases there is a
faint indication of bands, due very often to the chitin showing at
the joints, but there is no actual pale ornamentation. This is the
condition which is sometimes referred to as palpal banding in
A. nigripes, Staeger. A. immaculatus, Theo., also stated to have
banded palps, shows very indefinite and indistinct ones, not at all
sharply marked as in ordinary Anophelines.
Entirely unomamented palps are present in all Stethomyias,
including S. corethroides, Theo. (vide section on scale structure),
and S. aitkeni, James and Liston. Unbanded palps are also
commonly present in Anopheles , and occur in Mysorhynchus and
Cyeloleppteron. Otherwise the palps are usually ornamented.
In Anopheles the unspotted wing species also have unbanded
palps (except A. (?) smithi, Theo.*). The species A. gigas, Giles,
and A. simlensis, James, with banded palps have been separated
by James 1 as Patagiamyia. A. forntosus, Ludlow, since it is a hill
species whose description closely resembles that of gigas , would
seem also to be a Patagiamyia. A. punctipennis, Say., which is a
Patagiamyia (vide section on scale structure), has banded palps.
It would be very interesting to know what was the condition of the
prothoracic lobes in A. eiseni, A. crucians, etc. A. welcomei , Theo.,
is a peculiar species; it has no prothoracic tuft, but I am doubtful
if it should be considered as an Anopheles.
When the palps are ornamented, this is in the form of bands,
supplemented in some cases by extra spots of white (speckling) on
certain segments.
It is usual to talk of four or three-banded palps, etc., some of
the bands being described as broad or narrow, as the case may be.
The only accurate method is to refer to the position of the bands
in respect to the palpal segments. This relation is most easily
expressed in terms of leucogenetic centres. In spite of considerable
variety of appearance, a comparatively few general schemes of
coloration associated with different relative lengths of the palpal
* Note. —Thu species has a scale tuft on the prothorax and is therefore not an Anopheles
in the sense of James.
4 «
segments (vide section on structural variation) can be made to
account for the markings.
Scheme A. Palps with leucogenetic centres at each end of the
segments, but none of these actively spreading.
The effect of the appearance of the leucogenetic areas (following
upon complete absence of banding) in this scheme is the production
of four narrow pale bands, including the pale apex. If the species
be orthodactyloust, a somewhat different appearance is given to that
produced in the case of a markedly heterodactylous* species
(compare C. pulcherrima with the variety of P. nursei showing a
dark apical band. Plate V, figs. 8 and io).
This degree of extension of whitening is not exceeded in
Myzorhynchus, and is typical of palpal ornamentation seen in
Myzorhynchus, Cycloleppteron, Arribalzagia, Patagiamyia and
Cellia.
It seems to be a stable condition, and is but little liable to
variation in individuals belonging to these groups. In Myzomyia,
Pyretophorus and Nyssorhynchus this stage is unusual, and when
it does occur it is most often as an individual variation, e.g., the
four-banded palp varieties of N. fuliginosus, Ne. fowleri and
P. nursei. A permanent four-banded condition in this group is
seen in P. cinereus, Theo.
Scheme B. Leucogenetic centres at the ends of the segments,
with that at the end of the apical segment active.
In this scheme the apical centre extends, making the apical
segment entirely pale. This results in the ordinary type of three-
banded palp. Considerable difference in general appearance again
depends on whether the palp is orthodactylous or heterodactylous
(Plate V, figs. 11-13).
The narrow apical band in Myzomyia t ( culicifacies, funestus ,
etc.) is homologous in regard to extent of palp involved with the
broad apical band of Pm. rossi, but not with the narrow apical
band in Pi. simlensis or C. pulcherrima. The broad band in
• Vide Section III.
f Note.— As pointed out by Edwards 6 , Nyssorhynchus , Blanchard (= Laver ania, Theo.)
having as type species argyrotarsis , Rob-Dcsv., must take precedence over Cellia, Theo., a new
name being required for the group generally known as Nyssorhynchus. Again rossi , Giles, is
the type species of Myzomyia , Blanchard (= Grassia , Theo.) so that the name Pseudomyzomyia ,
Theo., is incorrect, whilst a new name is required for what is now generally known as
Myzomyia. In the present paper, however, the names in general use by Theobald arc
retained as being less confusing than new ones or old ones applied in a new sense.
49
Pm. rossi again is not comparable with the broad apical band in
N. macula/us. (Plate V, fig. 14.)
Certain species, as previously mentioned, which normally or
sometimes exhibit this condition (i.e., whitening of the whole apical
segment) may at other times show a black band, due to incomplete
extension of the apical pale area. The liability to this variation
is not merely accidental. In the case of Nyssorhynchus, those
species liable to the variation are distinctly related and exhibit
affinities to Cellia. In Mysomyia incomplete extension of the apical
white area is rare. It is more common in the heterodactylous
Pyretophorus , e.g., P. cinereus, as a variation in P. nursei, etc.
A few species, all apparently related, show a peculiar variant
of palpal banding, due to the absence of development of the apical
pale area. These dark-tipped species which show three bands are
M. turkhudi, Liston, M. Hispaniola, Theo., P. myzomifacies , Theo.,
and P. chaudoyei, Theo.
In M. turkhudi this character is liable to some variation, the
dark tip not being always very clearly shown.
SCHEME C. So far, the only leucogenetic centre that has shown
active spreading properties is that at the apex of the terminal
segment. Another type of palp is seen in which the centres at both
ends of the penultimate segment and at the apex of the second
segment also become active. In this case whitening spreads up and
down the shafts of the segments to produce the appearance seen in
N. maculatus (Plate V, fig. 14). Here the residual pigment area is
displayed as occupying almost the exact middle of the segment.
The apical white band in such a case is not comparable with what
looks like a very similar band in the case of Pm. rossi, etc.
Variation here usually consists in a partial, or more rarely in a
complete effacement of the pigment centre on the penultimate
segment.
Scheme D. What seems to be essentially a distinct scheme of
ornamentation to any of the above, is one which may be described
as showing in each segment only one apically situated leucogenetic
centre and a basal ly situated residual pigment area. Species
showing this type of palpal ornamentation are chiefly included under
the many-spotted-winged species of Donitz, i.e., M. punctulata,
Nm. elegans, N. annulipes, etc.
So
The scheme is beautifully demonstrated in Ch. kochi, Donitz,
and it is apparently that on which the palpal ornamentation of
N. karwari , James and Liston, is based (vide Plate V, fig. 15)- The
condition is very distinct from that ordinarily seen in Nyssor-
hynchus.
Possibly other variations or schemes of palpal ornamentation
exist, but enough has been said to show the nature of the condition
present in the great majority of species.
Markings of the legs. The occurrence of pale areas at the
terminations of the femur, tibia and first tarsus is usual, and such
may be conveniently termed ‘ knee spots.' Except as an unusual
variation, knee spots appear to be absent only in Stethomyia.
The occurrence of pale areas at the termination of the tarsal
segments gives rise to an appearance which may be called tarsal
banding. Tarsal banding may be of several kinds, depending upon
whether the banding is a mere trace not very definite and confined
to the actual joint, or whether it is broad and distinct involving an
appreciable extent of the ends of the segments. In some cases,
though banding is narrow, it is (as in P. jeyporensis) very distinct.
In the case of broad banding, there are again two conditions
depending upon whether the light area is merely somewhat paler
than the rest of the segment or is pure white. The latter condition
is, of course, much more conspicuous.
In some species the tips of the hind legs (and very rarely those
of other legs) are white. This may be a condition in which there
is a large continuous white area (as in N. fuliginosus, My. paludis,
etc.), or one in which the whole tarsus shows alternate dark and
light bands, one of which includes the tip. In the former case it
will be found that two or three segments are completely devoid of
any dark scales. In the latter case, though the terminal segment
may be all white, the other segments still have some portion of
their extent dark. In some cases, though the tip is white, not even
one segment is altogether white, the terminal portion of white
consisting in this case, as a rule, of a half or less, of the last tarsal
segment. In separation of species the exact details regarding the
number of segments white is important, but in the present paper,
dealing somewhat broadly with the subject, I shall refer to two
conditions only, i.e., (a) a continuous area of white involving at
5 1
least two segments; (b) an interrupted black and white appearance
of the tarsus in which the tip is white (i.e., as in N. maculatus,
Theo.).
In the case of the tarsus of certain Culicinae there may be a
development of pale areas at both ends of the segment or at one
or other end only, so that there results apical and basal, apical, or
basal banding as the case may be. Seeing how important the
coloration scheme appears to be, such distinctions may have a good
deal more systematic importance than one might be ready to grant
at first sight. So far I have not sufficiently studied the nature of
broad tarsal banding, and for the present shall not lay any stress
on differences in this respect. In many species, however, the
banding is purely apical, whilst in Pm. rossi, for example, it is
distinctly apical and basal.
The condition called speckling is one in which distinct spots or
bands are produced on the tibia or femur. Under an objective
a certain amount of admixture of dark and light scales is often
seen, and this may lead to a ‘ mottled ’ appearance under a lens.
This is quite distinct from the defined spots and bands referred to
as speckling. Speckling signifies a high degree of ornamentation
and is usually associated with tarsal banding and, very frequently,
with a white tipped tarsus.
An absence of speckling is characteristic of Myzomyia and of
the superpictus group of Pyretopkorus. Speckled species of
Pyretophorus seem to be in many ways distinct from the latter
group. Note, for example, P. ardensis, Theo., with ‘furry’ palps
resembling those of a Nyssorhynchtts, and the highly ornamented
P. aureosquamiger , Theo. Vide also remarks on the scale structure
and palpal characters of P. costalis.
In Nyssorhynchtts speckling is the rule, and reaches a high
degree of perfection. In Anopheles, Myzorhynchus and
Patagiamyia, speckling is absent, but it is developed in
Cycloleppteron, evidently a specialised but closely related genus
to Myzorhynchus.
Ornamentation of the wing in Anopheles. The study of wing
markings is complicated by the number of veins which may bear
spots, the fact that the arrangement of spots on each vein may
vary and that combinations are possible.
52
II. ORNAMENTATION OF THE WING IN ANOPHEUNAE
As is well known, only the veins in mosquitos carry scales, and
any spots that may be present are, therefore, restricted to these
structures, and are linear in shape.
In this respect the spots chi Anopheles differ from those in
butterflies, not only in being linear, but in the fact that the spots
in butterflies, as has been shown by Mayer, 2 originate on the wing
between the veins, and only involve the veins by extension.
In the Nematocera two types of spots are seen (a) dark spots
situated around vein junctions, and ( b ) dark wavy bands with
serpiginous edges which extend across the wing. These often look
as if their wavy margins had been caused by encroachment of pale
areas developed between the veins.
The former type of spots are constantly encountered. On
page 92, for example, of Theobald’s Monograph, Vol. I, is figured
a wing of Rhyphus in which dark patches are seen (1) at the
junction of the subcosta with the costa, (2) at the junction of the
first and upper branch of the second with the wing edge, (3) at the
termination of the lower branch of the second, (4) at the cross
vein linking the second with the next vein, and at two other
junctions.
The most striking development of pale spots forming between
the veins, and dark spots in connection with vein junctions I have
seen has been in the case of certain species of spotted-winged
Chironomus . In these species both wing membrane and vein is
covered with minute hairs not unlike scales, and so the condition
approaches that seen in Anophelines, whilst it seems to show what
the primitive condition may have been. In these wings the two
conditions seen are (1) aggregations of the hairs (or scales) in
dense patches in the neighbourhood of the main cross veins and
at two areas which seem to represent the bifurcation of the second
and fourth longitudinal veins, and (2) distinct pale eye-like spots
which occur on the wing membrane and have the minute scales
covering them also pale (Plate VI, fig. 4). In one species examined
there was a complete series of such spots occupying the cells and
ranging along the whole outer and posterior aspect of the wing.
I have described these conditions at some length because
53
1 believe they help one to understand certain of the facts to be
described in the case of Anophelines, and because they seem to show
so distinctly the primitive character of scale aggregation at the
cross veins, and the actual existence of something very like my
leucogenetic centres.
Primitive spotting in Anophelinae. The spotting seen on the
wings of Anopheles is of two kinds:
(a) Spots due to special aggregations of scales;
(b) Spots due to alternate areas of light and dark scales.
The first type of spotting is very familiar in the case of the
wings of A. maculipennis, Meigen. In this species no pale scales
are present on the wing veins, but there are aggregations of scales
forming dark spots. Such aggregations occur at the origin of the
second longitudinal, at the origin of the third longitudinal, to a
less extent on the neighbouring parts of the fourth and fifth veins
(i.e., in the neighbourhood of the cross veins), and at the bifurcation
of the second and fourth veins. This type of spotting is
conspicuous in A. maculipennis , because the wings are devoid of
other markings; but it is by no means confined to this species or
to species with unspotted wings, and it pccurs through a wide
series of Anophelines, often along with considerable ornamentation
of the second type of spotting. As it is seen chiefly (if not entirely)
in what I shall show reason to believe are certain primitive groups,
and as it is quite distinct from the spotting due to pale and dark
scales, and as something very like it occurs in other groups than
the Culicidae, I have called this primitive spotting. All species of
Myzorhynchus which I have examined show it, also Cycloleppteron
grabhamii, Theo. It is very plainly to be seen in Pt. lindesayi,
Giles, and is prominent in L. asiatica, Leicester.
The species P. atratipes , Skuse, has an unspotted costa, its palps
are unbanded, and it shows very marked and prominent primitive
spotting. It clearly cannot be a Pyretophorus, and the Certainty
with which one can speak is a good example of the value of colour
markings in systematic work upon the Anophelinae.
Scale clusters. In Myzorhynchus and Cycloleppteron there is
a condition allied to this primitive spotting, in which dark spots
on the veins are formed by scales larger than those on the light
54
areas. Such areas of large scales are seen, especially on the sixth
vein, forming the two dark spots characteristic of these genera.
Representation of spots on the lower surface of the wing. The
veins of Anophelines carry scales on both surfaces. The character of
the scales carried on each depends, however, on the particular vein
concerned. In all Anophelines that I have examined, the upper
surface of the first, third, fifth and sixth longitudinal veins carries
blunt appressed, and what may be described as flat scales. On the
under surface, all these veins (where they have scales at all) carry
quite a different kind of scale, which in most Anophelines is pointed,
and projects freely from the vein. The arrangement of scales
carried by the second and fourth vein is reversed, flat scales being
present on the under and projecting ones on the upper surface.
What have been called lateral wing scales are, for the most part,
simply the projecting scales on one or other aspect of the vein, as
the case may be. As the end of the wing is approached, i.e., about
the level of the forks of the second and fourth, the flat scales
become increasingly long and projecting, so that in this position
both surfaces of the wing carry projecting scales. This accounts
for the appearance of the spots in this region which have a blurred
look.
Except for a patch of scales towards its free end, or at most,
over half its length, the sixth vein is devoid of scales on the lower
surface of the wing. The main fifth and the inner part of the
branch are similarly bare underneath. The result is that spots on
these veins are very distinct and, under an objective, appear to be
formed of special small flattened scales. The remaining veins carry
scales on both surfaces.
On the whole, the upper surface of the wing is the more heavily
scaled, and partly on this account, and partly owing to the
preponderance of flat scales, the colour markings are, as a rule,
more intensely represented on the upper than on the under surface.
In most Anophelines, allowing for the somewhat more intense
coloration of the upper surface and the scale characters noted, the
spots on the upper and under surface correspond. But in certain
groups there is a marked lack of pale areas on the under surface
corresponding with those on the upper, which may reach such an
extent that the under surface of the wing is practically dark scaled
55
throughout. A good test of this condition is the third longitudinal,
which is most frequently a pale vein. In species where want of
correspondence of coloration is marked, the pale, flat scales on the
upper surface of this vein will be seen to be flanked by dark lateral
scales (i.e., ventral scales), seen through the membrane.
This appearance, called admixture of dark and light scales, has
often been referred to, but, so far as I know, no one has previously
realised its nature or significance. The group in which dark scaling
of the under surface of the wing reaches its most striking develop¬
ment is the Myzorhynchus group, including Cycloleppteron and
Arribalzagia.
A rather interesting condition was noted in the halteres of a
specimen of Cycloleppteron medio punctatus, which showed,
distinctly, the appearance of an unexpanded wing. These
structures were covered with snowy white scales on the upper surface,
with a central dark spot, but on the under surface were clothed with
deep black scales.
The wing of Myzorhynchella nigra , Theo., is almost entirely
dark scaled beneath, though it has many pale spots on the upper
surface; also many pale spots present on the upper surface are
lacking on the lower in P. watsonii, Leicester {Myzorhynchus ?) a
species allied to My. (?) natalensis, Hill and Haydon, as well as in
Nm. elegans, James. Cellia squamosa, Theo., and M. lutzi, Theo.,
show a third longitudinal, light above and dark below. P. atratipes,
Skuse, is another species in which the wing is almost entirely dark
scaled beneath, though it carries quite conspicuous pale areas on
the second, fourth, fifth and sixth veins above.
In Pt. gigas and Pt. simlensis the markings are, for the most
part, reproduced on the lower surface of the wing.
In Myzomyia, Pyretophorus, Nyssorhynchus and in Cellia
pulcherrima , Theo, the wing spotting is equally represented on
both surfaces, though in some ‘ dark ’ Nyssorhynchus dusky scales,
not definitely dark or light, may be present beneath the third vein.
Admixture of dark and light scales. Apart from the effect of
admixture, due to the showing through of dark scales beneath the
wing membrane, there is in Myzorhynchus, Cycloleppteron and
Arribalzagia a true occurrence of mixed black and pale scales on
many of the veins. Such admixture of scales is not seen in the
5 ^
majority of Anophelines, and does not occur, so far as I am aware,
in Myzomyia, Pyretophorus, Nyssorhynchus or Cellia.
All the characters so far mentioned must be looked upon as
special to certain groups of Anophelines. What one may call the
more ordinary Anophelines show neither primitive spotting, nor any
degree of unequal ornamentation of the wing surfaces, nor
admixture of light and dark scales on the veins, and in them
spotting depends entirely upon the existence of definite areas of
pale and dark scales on the veins.
Another condition of possible importance, but concerning which
my observations are too imperfect to enable me to deal, is staining
of the wing membrane in the area of dark spots. This is greatly
exaggerated in Cycloleppteron mediopunctatus, but is seen in many
species.
Nodal Leucogenetic Centres of the Wing
The wings exhibit very clearly a system of leucogenetic centres
and residual pigment areas. The leucogenetic centres are of two
kinds, those associated with what may be called ‘ nodal points ’ of
the wing, and those which I shall call ‘ internodal.'
The nodal points in the wing of an Anopheline which have
relation to spotting are shown by the small circles in Plate VI,
fig. i. They may be enumerated as follows: —
(1) The apical nodal point at the junction of the first
longitudinal vein with the costa ( Ap .).
(2) The subcostal nodal point at the junction of the subcostal
vein with the costa ( S.c .).
(3) The humeral nodal point where the humeral cross vein enters
the costa near its base ( h .).
(4) Nodal points in connection with the cross veins ( C a -C 6 ).
(5) Nodal points at the bases of the forked cells (bifurcation of
veins two and four).
(6) Nodal points at the junction of all veins and their branches
with the wing margin.
There are other points whose nodal origin is possible or probable.
These are: —
(7) A point on the first longitudinal vein slightly internal to
57
the termination of the second longitudinal. If, instead of being
joined to the first longitudinal by a cross vein (cross vein 2), the
second longitudinal itself formed a direct junction with this vein
{Vide remarks on wing venation in Section III), it would presumably
enter at the nodal point in question. Some authors figure the vein
as acting in this way; but I have not found any example of an
Anopheline wing showing this arrangement. On the other hand,
it appears to occur in some other Culicidae (occasionally in the case
of Theobaldia annulata). The nodal point in question is an
important one, and, as it occurs at the origin of the Sector radius
vein, I have called it the Sector nodal point {S'). The nodal point
at the entrance of the cross vein actually linking on the Radio-
Sector, which is also a very important one in relation to wing
markings, may be called the Accessory Sector {S').
(8) It is possible that a nodal point occurs on the first longi¬
tudinal at C. in the diagram. In Anophelines, no sign of any
cross vein exists here, but such a cross vein is commonly seen in
Tipulids {Vide Plate VI, fig. 2). Another possible nodal point is
at C.V. Breaking of the subapical costal spot in P. costalis, and
some other species, also suggests a point of a nodal character,
marked with a (?) in the diagram.
These points behave as nodal points, and it is possible that a
study of the developing wing in the nymph may throw light on
their origin. But at present their structural origin can only be
suspected.
It is noticeable in regard to white areas formed at the nodal
points, that they do not show so active a tendency to spread as do
those of an internodal character.
Species showing ' dark * or * light ’ areas at the cross veins and
bifurcations. Certain nodal points may be represented by a
leucogenetic centre or by a pigment centre. The nodal points on
the costa rarely exhibit the latter condition, but the humeral junction
is not infrequently the site of a pigment spot (instead of a pale
spot), as in Cycloleppteron grabhamii. Still more rarely, the
junction of the subcostal vein may be the site of a dark spot,
Cy. mediopunctatus and Ar. maculipes, whilst a dark spot in close
association with this junction is seen in Nm. elegans, M. punctulata
and N. annulipes, Walker. Such spots form small accessory spots
on the costa (apart from the basal accessory spots to be described
later), and are possibly of much greater significance than their mere
size might suggest.
The nodal points of greatest importance in connection with their
power to be either ' dark * or ‘ light 1 are those at the cross veins and
bases of the cells. In the genera Myzomyia , Pyretophorus ,
Nyssorhynchus , Cellia , and Neomyzotnyia , the cross veins are always
the site of light interruptions. In Myzorhynchus , Patagiamyia ,
Cycloleppteron , Arribalzagia , and Myzorhnychella these areas not
only are dark, but, when given the chance, exhibit themselves as
residual pigment centres. In a great many cases, in addition, they
are the site of scale aggregations, giving rise to primitive spots
( Vide Plate VIII, figs. 1-13).
'Nodal points on the costa and first longitudinal . In those
species with unspotted costa, we may consider that none of the
nodal leucogenetic centres have displayed themselves. This is the
typical condition in Anopheles , restricting this term to the genus
as defined by James (i.e., without a prothoracic tuft). It is also
the condition in Stethomyia.
The first nodal leucogenetic centre to become evident in all other
species, without exception, is the apical , giving rise to the condition
seen in Pt. lindesayi and P. atratipes.
The next nodal centre to appear is invariably the subcostal.
This is the typical condition in Myzorhynchus , though in some
species {My. sinensis , My. pseudopictus) the sector spot has also*
made its appearance on the first longitudinal, but has not spread
on to the costa. The same condition is seen in L. asiatica and in
the related form My. wellingtonianus , Alcock.
The third nodal centre to appear as a pale spot is either the
sector spot or the humeral spot , usually together with a spot at the
extreme base of the wing and at the (? nodal) point C. I.'
If the sector spot becomes prominent without the others there
results the sinensis type of wing. If the sector spot remains in
abeyance and others develop, the gigas and punctipennis type of
wing is produced (Plate VIII, figs. 9 and 4).
So far, all species showing no further elaboration of the costal
spotting than that described, have dark cross vein areas, frequently
exhibit primitive spotting and a scheme of wing ornamentation in
59
regard to internodal spots, which is distinct from that seen in
Myzomyia, Pyretophorus, etc.
The sector spot and the (?) nodal spot at C. 1 ." appear in most
cases simultaneously, giving rise to a four spotted costa. The
condition in Cycloleppteron grabhamii is that shown in
Myzorhynchus. In the case of C. medio punct atus, the sector spot
has included the costa, pale areas have appeared at the base, and
the extent of the dark scaled portion is less. In Arribalzagia
maculipes the dark costal spots are still more sharply differentiated
and restricted. Both these greatly modified wings appear to be
distinctly a development of the Myzorhynchus pattern. In
Myzomyia, Pyretophorus, Nyssorhynchus, Neocellia, and Cellia,
the costal pattern, except as the result of occasional individual or
specific ( ?) variation, shows four main spots due to the formation
of pale centres at the apical, subapical, subcostal, and sector nodal
points, respectively. In the great majority there is also a pale area
at the humeral, at the base of the wing, and at C.iJ. The
accessory sector spot may be blended with the sector spot, merely
shortening a little the middle spot as represented on the first
longitudinal vein (e.g., M. culifacies). It may be lost in the
general extension of the sector spot (e.g., certain Pyretophorus'), or
it may form a distinct interruption towards the inner side of the
middle dark costal spot, as is the case throughout Nyssorhynchus.
In most Nyssorhynchus a very similar interruption is also seen
towards the outer end of the spot. The exact origin of this second
interruption is uncertain, but it lies opposite the junction of the
third cross vein with the stem of the second longitudinal. Where
the inner interruption only is present, I shall speak of the middle
costal spot as being * incompletely broken,’ where both interruptions
are seen, I shall refer to this spot as ‘ completely broken ’
(Plate VIII, figs. 34, 3;).
In certain species one or both of the small dark scaled portions
on the first longitudinal, separated off by the pale interruptions,
may have disappeared. The disappearance of both small spots
gives what is frequently spoken of as the ‘ T ’ shaped spot of
Pm. rossi.
The same kind of interruption may occur, but not commonly,
on the first longitudinal, under the subapical dark costal spot.
6o
This is very characteristic of P . costalis , and is seen particularly
in the many-spotted winged species. It can be referred to as a
broken subapical spot. (Plate VIII, fig. 25.)
Accessory costal spots . On the costa at the base of the wing
are very commonly seen from one to three small dark spots which
as a rule are unrepresented on the first longitudinal or even on the
subcosta. These may conveniently be termed basal accessory spots .
In Myzomyia , Pyretophorus , Nyssorhynchus , etc., two such spots
are most commonly seen lying on either side of the junction of the
humeral cross vein; but three are sometimes present, and not
infrequently they may join together among themselves, or, either
in part or wholly, be linked on to the inner main costal spot, giving
the appearance commonly seen in M. listoni. (Plate VIII, fig. 28.)
Another type of basal accessory spot is sometimes seen where
the dark area occurs at the junction of the humeral cross vein,
e.g., Cycloleppteron mediopunctatus .
The number and arrangement of accessory basal spots, or
whether any light areas exist at all on this part of the costa, does
not seem to be very important.
The selection of the site of junction of the humeral vein for a
dark spot is, however, very significant.
Accessory costal spots other than the basal accessory spots are
found in certain cases. These appear to be of considerable
significance, and their exact site is very important. The mode of
formation and homology of such spots in different species can be
traced. In Ch. kochi y Donitz., a prominent accessory spot is seen
on the costa between the inner and middle dark costal spot. The
nature of this spot is clearly shown by a study of the wings of
P. watsonii , Leicester, and of My . natalcnsis> Hill and Haydon.
The wing of My (?) natalensis , as shown in the photograph given
by its describers, is a beautiful demonstration of the typical sub¬
division of the costa into dark spots by the appearance of the
apical, subcostal, sector, accessory sector and C. I." nodal leuco-
genetic centres. In the species P. watsonii , Leicester, the type of
which is in the British Museum, a very similar arrangement of
spotting is shown, but the accessory sector spot has developed to
such an extent as to reach the costa, forming a small detached dark
spot clearly identical with that seen in Ch. kochi . This fact is the
6i
more interesting in that such a spot is not common in Anophelinae,
and that it is for the most part seen in the many-spotted-winged
species, e.g., Nm. leucosphyrus , Donitz. P. watsonii and My.
natalensis show therefore very curious affinities; showing an
approach to Myzorhynchus they so far differ as to have been
excluded, or to have been regarded as doubtfully belonging to
this genus, by their describers; they distinctly recall some
Patagiamyia characters and as pointed out they have some unusual
points in common with the many-spotted-winged group and with
Ch. kochi , Donitz, which species it should be remembered has
Myzorhynchus- like wing scales.
Bridging of costal spots on the first longitudinal. Certain
species, including C. squamosa , Theo., C. albimana , Wied., and
N. fuliginosus , Giles, show a bridging of certain of the pale costal
areas by dark areas on the first longitudinal. The same feature is
displayed in Myzorhynchella nigra , Theo. The species Myzorhyn -
chella nigra shows a four-spotted costa; it has a wing with a fair
number of markings above, but almost entirely dark-scaled below;
the cross vein nodal points are dark; it has prothoracic tufts; there
can be little doubt but that it is related to Myzorhynchus. If, as
seems possible, C. squamosa , which exhibits distinct Myzorhynchus -
like affinities, really descended from a primitive Myzorhynchus-\\kz
ancestor, one would expect this form to be something like
Myzorhynchella> though the evidence for such a descent, and for
that tentatively suggested for the elegans group, is very slender.
■»
Nodal points on wing margin. When pale spots display them¬
selves in this situation they affect not only the veins but also the
wing fringe, forming ' pale fringe spots.’ The amount of possible
spread of these spots, like most other spots of nodal origin, is
small.
Internodal Leucogenetic Centres of the Wing
The internodal spots appear as a rule almost exactly in the
centre of long stretches of dark vein lying between nodal points,
and the order of appearance of the internodal spots has a distinct
relation to the length of the dark area concerned. The internodal
points in the scheme of coloration shown by Myzomyia, Pyreto-
phorus, Nyssorhynckus, etc., are: —
62
(1) One on the upper branch of the second (2 1 ). Two on the
lower branch (2 2 and 2 3 ). These are usually so placed that they
alternate, as shown in the diagram, and the result is that' the limbs
of this forked cell are asymetrically marked.
(2) On the shaft of the third vein. Whether one or two centres
are here concerned is not quite certain, but I am inclined to think
that there are two which very readily blend (vide appearance shown
in some species).
(3) One on the upper branch of the fourth vein (4 1 ). One on
the lower branch (4 2 ). In this case the branches are marked
symmetrically, though owing to the slope of the wing edge the
actual position of the spots is not quite opposite one another.
(4) One on the upper branch of the fifth, i.e., between the
nodes C. 5. and 5'. Two on the main branch of the fifth, one above
and one below the junction of the branch.
(5) Two on the sixth, dividing this vein approximately into
three portions.
Not only have these internodal centres a nearly fixed position,
but they have an almost fixed order in which they become apparent
by the development of pale spots.
The internodal spots, once they appear, tend to extend very
rapidly along the veins, causing much more extended pale areas
than do the nodal points.
The first spot to appear is 5 1 on the main fifth beyond the fork.
In certain species this is the only internodal spot present (e.g.,
funesta var. umbrosa ). In M. culicifacies , M. nili t etc., this is also
the only conspicuous spot present.
The next spot to appear is generally one of those on the third
vein, giving rise to a variety of Af. culicifacies and M. funesta var.
subumbrosa , with more or less pale third longitudinal. In association
with this appears a spot on the upper portion of the fifth (5 2 ), and
a pale area at the base of the fourth longitudinal (4). The wing at
this stage of ornamentation is that seen in the lighter-winged
Myzomyiasy i.e., M. listoni y M. funesta , etc. The only Myzomyia
(excluding certain forms previously mentioned) which shows any
further ornamentation is M. albirostris , Theo. (M. aconita , Donitz ?),
a peculiar species in many respects, including the larval characters.
With a further degree of lightening of the wing, additional spots
appear at 5 2 on the branch of the fifth, and on the forked cells,
2 3 being a little later in appearance than the others. Whilst these
areas have, as it were, only just appeared, the ones previously noted
have spread considerably, so that as a rule the third vein is
completely pale, for example, in any species showing a pale spot
at 5 2 .
The scheme so far developed is that seen in a number of species
of Pyretophorus; though in other species of this genus an additional
spot (6 2 ) has appeared.
The nodal points on the edge of the wing behave in concert with
this progression. In the wings showing only 5 1 the fringe spots are
generally deficient (e.g., two in M. culicifacies). In association
with the appearance of the nodal centres 3 and 5 2 , fringe spots
are generally found at all veins except the sixth. The nodal point
at the termination of this last vein only occurs as a rule after 6 2 has
appeared. At about the same time another (?) nodal spot not
previously represented forms a pale area on the first longitudinal,
opposite cross vein 3. It is this which gives the double break in the
middle costal spot so characteristic of Nyssorhynchus , etc.
Whilst no further intemodal spots appear, the wing still
continues to show modifications owing to the encroachment upon,
and in some cases obliteration of, residual pigment areas. The
more important changes produced by this extension of the internodal
centres are {a) the blending of 5 1 and 5 2 , thus producing the
commonest condition of the fifth vein in Anophelines (vide Plate
VII, fig. 4); ( b ) the obliteration of one or both of the small spots on
the first longitudinal under the main spot (points of the 1 T ’ in
rossi ); (c) obliteration of one of the dark spots on the sixth vein,
either the upper or the middle one as in some species of Cellia\
( d) obliteration of one or more of the small dark spots on the forked
cells (usually the first to go is 2" b , the next most usually 2' b or
4* b ); (*) the obliteration of the dark spot 4 r.
In addition to mere extent of white involved there is a difference
in the intensity of the colour. Roughly speaking, Myzomyia and
Pyretophorus show pale but not markedly white areas. Nyssorhyn¬
chus and Cellia exhibit a striking dead-white ornamentation.
In Myzorhynchus , Cycloleppteron y Patagiamyia , etc., some of
the intemodal centres appear to be the same as certain of those
6 4
described, but there is no general correspondence in the scheme of
coloration. Also in this group there seems to be a much less orderly
sequence of such pale areas as do appear. The whole effect of the
ornamentation is of course modified by the fact that the cross vein
nodal centres are dark.
A group showing light nodal centres, but having a wing
ornamentation scheme absolutely without relation to that of
Myzomyia , etc., is the group Donitz 4 has drawn attention to as
markedly peculiar owing to the character of the wing spotting. The
most striking character of the wing in this case is the extraordinary
multiplication of spots due to alternate short lengths of dark and
light scaling on the veins. Practically none of the intemodal areas
seen so regularly throughout Myzomyia , Pyretophorus > etc., can
here be made out. The sixth vein shows from four to six dark
areas, the main fifth at least three interruptions on each portion of
the stem and on the branch respectively. The branches of the
second and fourth veins have each a double interruption. The third,
instead of being throughout most of its length either dark or pale,
is broken up into alternate light and dark areas.
Summarising what has been detailed above, we can say that
there are three absolutely distinct colour schemes on which the
spotting of wings of Anophelines is modelled. These are: —
(1) Scheme A. The scheme on which the wings of most ordinary
Anophelines are spotted, characterised by the very regular
appearance in a definite sequence of certain intemodal pale spots
and by the cross veins and bifurcation nodal points being pale.
(2) Scheme B. A less regular scheme shown by Myzorhynckus ,
Cycloleppteron , Patagiamyia , etc., in which the nodal points at the
cross veins and bifurcations are dark.
(3) Scheme C. A scheme characterised by multiple spotting of
the veins and the occurrence of more than three dark spots on the
sixth.
These three schemes not only seem of the most fundamental
importance, but as displayed in the great majority of species of
Anopheles can be distinguished at a glance.
65
Variation in Colour Marking shown by Individuals of the same
Species
In any attempt to utilise colour markings as a basis of classifica¬
tion the question of individual variation must be considered. It
may be objected that such variation as is displayed in many species
will prevent use being made of colour markings. This I think is
not necessarily so, because variations shown by individuals of the
same species have only a certain latitude, and with a correct
appreciation of what a given variation really amounts to one is able
to allow for variation in classification. The appearance of a new
centre out of its order, or the change of the markings over several
steps of normal colour marking development, would be a much more
serious matter than the final extinction of a disappearing pigment
area or the appearance of a pale centre already on the point of
appearing. In this respect I believe a knowledge of the principles
underlying colour marking will give an added importance to
observations upon individual variations. Certain stages seem
distinctly unstable, others much more stable. The knowledge as to
the relative stability of different conditions cannot but be of value
in deciding whether certain forms are species or varieties.
III. VARIATION IN STRUCTURE OTHER THAN THAT
CONNECTED WITH SCALES
Very few structural variations other than those connected with
scales are mentioned in descriptions of species of Anophelinae. This
is in itself rather suggestive that such variations as do occur are not
of an obtrusive nature. So far as I can ascertain the following are
almost the only points in which different species of Anophelines
differ from one another in respect to structure.
General shape of body. Donitz 4 has called attention to certain
characters of the head and the shape of the thorax in Anophelinae.
These require fresh material for proper study, and my observations
in this respect do not add anything to what was previously known.
Closely associated with the general shape of the body is the
attitude adopted by the species when at rest. The most importance
attaches to the possession of a ‘ culex ’-like attitude. This attitude
seems to be a definite character of Stethomyias. M. culicifacies,
66
Giles, and Chagasia fajardoi, Cruz, are the only species apart
from this group which, so far as I know, have been described as
adopting this attitude. The ‘ culex ’ attitude in the former is not
nearly so striking as in the case of S. aitkeni, which superficially
almost exactly resembles some long-legged Aedine mosquito.
Apart from the assumption or not of the ‘ culex ’ attitude, there
are marked differences in the angle made by different species with
the surface on which they are at rest. Observations in this respect
are too limited to allow any general conclusions being drawn as to
any possible significance attitude may possess in respect to generic
affinities.
The antennae. In the female antenna the arrangement of the
segments in respect to their length is somewhat peculiar. Following
upon the large globular basal (second segment according to Nuttall)
is a rather long segment. The next segment is in most species not
only the shortest in the whole antenna, but is often almost or quite
globular. In My. barbirostris, My. umbrosus, My. paludis.
My. sinensis and C. grabhamii, though this segment is still the
shortest in the antenna it is not nearly so disproportionately
shortened, nor has it the same globular appearance as in most
Anophelines. The segment is also rather longer than usual in
P. jeyporensis, James.
The male antenna possesses a very striking structural
peculiarity in the great length of the two terminal segments. But
so far as I could see the relative lengths of these structures
exhibited very little change in different species, nor do the antennae
of different species differ noticeably in other respects.
Female palpi. The number of articulated segments in the
female palps in Anophelines is four. There is, however, a vestigial
prominence carrying tufts of scales at the extreme base of the palps
which probably represent a fifth segment.
Distinct variations occur in the relative length of the different
segments. In this respect two types of palps can be distinguished,
which I have termed respectively orthodactylous and heterodactylous.
In the first or orthodactylous palps the segments observe a kind
of proportionate decrease in size as the apex of the organ is
approached. In the very orthodactylous genera, such as Myzorhyn-
chus , the terminal segment may be as much as two-thirds the length
67
of the penultimate. One may define an orthodactylous palp as one
in which, whilst the segments one and two keep their proportionate
length (they only vary to any extent in Stethomyia), the last segment
is not less than half the length of the penultimate segment.
In the second or heterodactylous type of palp there is a marked
want of regularity in the decrease exhibited by the segments. Of
heterodactylous palps two distinct types are encountered, due to
the disproportionate length of the preapical and second palpal
segment respectively.
Great increase in the length of the preapical segment is seen in
both Myzomyia and Pyretophorus, the apical segment being a third
or even a quarter of the preapical (Plate V, fig. io). Such a marked
structural variation in a sub-family otherwise showing very few
changes in structure must have considerable significance. Species
with distinctly orthodactylous palps now included in these genera,
e.g., P. costalis, Loew., would seem to require separation.
The second type of heterodactylous palps occurs in Stethomyia.
In this case the second segment is very long and the third
disproportionately short (Plate V, fig. 16). This variation is the
more remarkable in that amongst other Anophelines any marked
change in the relative length of the first and second segments is
conspicuously absent. This type of palp cannot be overlooked, if
attention is directed to it, and it would seem in itself a sufficient
and very good generic character. As previously mentioned,
S. corethroides shows the linear head scales characteristic of
S. ai/keni. It also shows the peculiar modification of the palps
mentioned above. A similar condition of the palp is figured by
Theobald for S. nimba.
Expressing the relationship of the terminal to the penultimate
segment in a decimal, one can obtain, when dealing with
orthodactylous palps or with the first type of heterodactylous palp,
a very useful index. What may be called the palpal index for a
number of species is given below. This index can be determined
very accurately from balsam-mounted specimens by direct measure¬
ment with an eye-piece micrometer, but better by making camera
lucida drawings, taking care to avoid any possibility of distortion.
Those indices given in the table where no other remark is made have
been determined in this way and checked by direct measurement.
68
Species
Palpal index
Source of information
My. barbirostris .
o-61
Mounted specimen
My. umbrosus .
0*69
My. mauritianus .
°*55
n
Cy. grabbamii .
0-70
11
A. maculipennis .
o -57
11
A. bifurcatus .
0-50
»!
Pt. lindesayi .
071
11
M. fumsta .
032
11
P. uursei .
0*32
11
P. cleopatrao .
o -35
11
P. jeyporensis .
0*32
11
Pm. rossi .
0*50
11
P. costalis .
075
N. fuliginous .
0*60
11
Ns. fowleri .
0*62
N. maculatus .
0*50
C. pulcberrima .
0*54
C. squamosa .
o-68
>1
C. olbimoua .
079
Specimen not mounted
M. aconita .
o -43
From Donitz's figures
M. btbos .
o -33
n
M. Acceptor .
059
11
Nm. leucospbyrut .
o*68
n
M. punctulata .
0*63
i»
M. tenebrosa .
o *73
i>
N. maculatus .
0-50
11
N. leucopus .
0*46—o-i7
”
P. ctnereus .
o -37
Photo by Hill Sc Haydon
P. ardensis .
075
»>
My. natalensis .
076
11
69
Given a good photograph, one can usually make a rough
determination of the palpal index; very often, however, a glance
will tell one whether a particular palp is orthodactylous or distinctly
heterodactylous.
In the drawings given in the Plate the palps of different species
have been drawn by means of a Zeiss adjustable lens ( a*), so that
they are shown of the same size (not scale). This shows very clearly
the variations in the proportions of the different segments. The
extremely low index of Mysomyia and the nursei group of
Pyretophorus is practically diagnostic.
The male palpi. These show a change more or less analogous
to that seen in the female, the palp being more abruptly clubbed
and the last two segments taken together measuring proportionately
less in the heterodactylous species than in others.
The wings. On the whole the structural characters of the wings
are more interesting from their fixed character than for the variations
they exhibit. Two conditions have, however, been made use of as
variables by Theobald and others. These are the positions of the
cross veins and the length of the forked cells. On these subjects I
have nothing new to add.
Much more important from our present point of view are certain
points connected with the homology of the wing venation in
Anophelines. According to Comstock and Needham the generalised
insect pattern of wing venation shows the following main
longitudinal vein systems, the branches of which are represented
more or less closely by tracheal branches which precede them in
development: —
(1) The costa.
(2) The subcosta.
(3) The radial system.
(4) The medial system.
(5) The cubital system.
(6) Three anal veins.
The subcosta is normally a bifurcate vein, the branches of which
enter the costa at two points. In many forms the lower branch
becomes fused with the radius for a portion of its length, so that
there appears to be (1) a branch joining the terminal portion of the
1 *
subcosta with the radius, and (2) a branch going from the radius to
the costa. The auxiliary or subcostal vein in Anophelines is an
unbranched vein entering the costa a little beyond its middle. It
seems quite clear that it is homologous with the subcosta of the
generalised pattern, and it shows the humeral cross vein joining it
and the costa near its base, which is a feature of the generalised
wing.
The radius in its typical development consists of an unbroken
vein R lt passing from the base of the wing to the neighbourhood of
the apex (Plate VI, fig. 2). Posteriorly to this there arises a branch
(the radio-sector) which sub-divides into two branches R a + ] , and
R 8 + 4 , each of which again sub-divides, forming four terminal
branches R a , R 3 , R 4 and R a . Between the lower branches of the
radio-sector (R 415 ), and the medial system is the second of Comstock
and Needham’s cross veins, the radio-medial. Nuttall and Shipley
consider the first longitudinal vein to be R 2 , the branches of the
second longitudinal to be R a and R 3 respectively, whilst the third
longitudinal is the combined R 4 + 5 . Following this homology the
radio-sector will be that portion of the second longitudinal between
its origin and the origin of the third vein, i.e., where the
supernumerary cross vein enters it.
Some figures show the second longitudinal as a branch of the
first. In reality, though it is joined near its origin to the first
longitudinal by a cross vein, the vein itself continues past this a
short distance to end on the wing membrane, as noted by Nuttall
and Shipley. Presuming the second longitudinal to be homologous
with the radio-sector, its true line of departure would be a little
internal to the cross vein, a point we have referred to when
discussing the formation of wing spots.
The division of the radio-sector into R t + t and R 4 + * would,
following Nuttall and Shipley’s homology, occur at the so-called
supernumerary cross vein. Some authors figure the third vein as
arising directly from the second, but this is not actually the case,
and the junction is brought about by means of a cross vein, the vein
itself continuing slightly beyond this to lose itself on the wing
membrane.
The cross vein joining the third with the stem of the fourth vein
would appear to be one of Comstock and Needham’s original cross
veins (radio-medial). The fourth longitudinal would appear to
represent the median, the fifth longitudinal the cubitus, and the
sixth the first anal.
The only cross veins in Anophelines actually represented as cross
veins in Comstock and Needham's generalised nomenclature are
(i) the humeral, (2) the medio-radial, (3) the radio-cubital, these
being respectively the humeral, the mid and posterior cross veins of
Theobald. But there are, as we have seen, additional cross veins
which are quite indistinguishable structurally from the first-
mentioned, and one of these (that linking the third with the second
vein) is called the supernumerary by Theobald. The cross vein at
the origin of the second vein, though noted by Nuttall and Shipley,
has not received a name. Since it links up the radio-sector to R x it
might appropriately be called the radio-sector cross vein. I would
suggest, however, the use of the terms first, second, third, fourth
and fifth as applied to the cross veins which makes no attempt, as
is very imperfectly done at present, to denote their homology. It
seems absurd to term the longitudinals by numbers and the much
less important cross veins by a hybrid nomenclature only partially
descriptive. The omission of the radio-sector cross vein, which is
every bit as important as the others, is also absurd. In the following
pages I shall refer to the cross veins according to the following
plan, which has the merit that each vein has the same number as the
longitudinal vein immediately behind it.
Homologous
nomenclature
Present
nomenclature
1
Suggested
name
Position
Humeral
Humeral
Humeral or first
Joins costa to subcosta
at base
(Radio-sector)
—
Second
Links on second vein
(Lower radial)
Supernumerary
Third
Links on third vein
Radio-medial
Mid-
Fourth
Joins fourth vein to
third
Medio-cubital
Posterior
Fifth
Joins fifth to fourth
On the system suggested the cross veins entering any given vein
can be at once named, or the veins that any cross vein affects be at
once called to mind without effort.
7 *
IV. STRUCTURAL VARIATION IN THE IMMATURE STAGES
A considerable number of larval forms have now been described,
and it seems possible to make some tentative use of the facts
gathered in attempting to ascertain the affinities of species. In
particular the variations in the antenna, i.e., whether it carries a
branched hair or not, and the occurrence in certain species of the
lanceolate type of palmate leaflet would seem to be significant.
Variations in the frontal hairs and other features have been
described for a number of species, but the application of such
variations to classification is less obvious.
Species with a branched antennal hair. So far as I have been
able to ascertain, the following species possess a well marked or
vestigial branched antennal hair.
A. maculipennis, Meig. Noted by Hill and Haydon.
A. bifurcatus, Linn. Vide Plate.
Pt. lindesayi, Giles. Stephens and Christophers.
A. crucians, Wied. Smith.
Pt. punctipennis, Say. Smith.
Pt. simlensis, James. James.
My. sinensis, Wied. Stephens and Christophers.
My. barbirostris, Van der Wulp. Stephens and Christophers.
My. paludis, Theo. Hill and Haydon.
Chagasia fajardoi, Lutz. Shown in drawing of larva by Silva.
The larval antenna of Cy. grabhamii, Theo., is figured by
Theobald without a branched hair. This structure can, however,
be very readily overlooked if attention is not especially directed
to it, and the observation requires confirmation. It is interesting
to note that no branched hair is present in My. natalensis, Hill and
Haydon.
Species with lanceolate type of palmate hair leaflet. The type
of leaflet seen in Mysorhynchus is not uncommon. The following
species have been described as showing lanceolate and serrated
palmate leaflets.
A. maculipennis, Meig. Hill and Haydon.
A. bifurcatus, Linn. Vide Plate.
My. barbirostris. Van der Wulp. Stephens and Christophers.
My. sinensis, Weid. Stephens and Christophers.
73
My. pdludis, Theo. Hill and Haydon.
Cy. grabhamii, Theo. Theobald.
C. albitnana, Wied. Graham.
C. squamosa var. arnoldi , Steph. and Christop. Newstead and
Carter.
C. albipes, Theo. Low.
Forms showing an approach to this type of leaflet, but in which
the leaflet is drawn without serrations and as of a simple fusiform
shape.
C. squamosa, Theo. By Hill and Haydon.
Mysorhynchella nigra, Theo. Silva.
Myzorhynchella parva, Chagas. Silva.
Stethomyia culiciformis, James. James.
Forms showing an approach to this type of leaflet in the
serrations at the shoulder of the leaflet are more drawn out than in
ordinary Anophelines has been noted in: —
My. (?) natalensis, Hill and Haydon. By these authors.
Pt. lindesayi, Giles. Stephens and Christophers.
5 . aitkeni, James. By this author.
Mm. elegans, James. By this author.
P. ardensis, Theo. Hill and Haydon.
Hill and Haydon figure N. pretoriensis with leaflets of this
character, but in the photograph accompanying their paper the
condition is not so clearly shown.
The extreme degree of branching of the external frontal hairs
characteristic of My. sinensis is also shown by My. paludis (Hill and
Haydon). This character may therefore be reasonably taken as
one of the features of Myzorhynchus. Its absence in any given
species would draw attention to doubtful generic position. Marked
feather-like branching resembling that in N. fuliginosus, Giles, is
also seen in C. squamosa and in C. pulcherrima. The following
tabular statement gives, as far as the information available allows,
the chief larval characters of a number of species arranged in the
order of their apparent phylogenetic significance.
Variations in the nymphal structure of Anophelines. Very little
variation can be detected in the nymph of different species. There
is, however, a slight but appreciable difference between the nymphal
trumpets of My. sinensis, My. barbirostris, Pt. simlensis,
A. maculipennis and A. bifur cat us and those of Anophelines
Table showing larval characters in order of apparent phylogenetic significance
2 i
§1
g -
6 II
So
if
!:
Palmate leaflets lanceolate
+ 4 ;
showing tendency as in
lindesayi 4-
Palmate leaflets fusiform.
Palmate leaflets with
ordinary longish filament.
Palmate leaflets with
stumpy filament,
External frontal hair
branched as in Sirumii B,
as in fuligitwus bb, other¬
wise distinctly branched b,
simple or with fine lateral
branches S.
Stctbowyia aitkeni
,, culiciformis .
o
o
4-
4
s
s
Anopheles maculipennis
+
f 4
B
,, hi/urcatus .
+
f 4
S
„ eiseni
+
?
s
crucians
+
?
s
Patagiamyia lindesayi .
+
4
s
,, simlensis ...
+
?
s
„ punctipennis
+
?
s
Myzorbyttcbus sinensis ...
+
4 4
B
,, barbirostris
4-
4 4
B
,, paludis .
+
4 4
B
Cycloleppteron grabbamii
?
4 4
S
Cbagasia fajardoi .
f-
4
Myzorbyncbus (?) natalensts
o
4
b
Neomyzomyia elegans ...
o
4
S
Cellia squamosa var. amoldi ...
o
4 4
bb
„ squamosa ...
o
4
bb
„ albimana .
o
4 4
,, albipes .
o
b
„ pulcberrima .
o
4
bb
Pyretopborus ardensis .
o
4
Myzorbyncbella nigra .
4
.. f arva .
4
Nyssorbyncbus fuliginosus
o
bb
„ nivipes .
o
4
bb
L
„ jamesi .
o
4
D
,, maculipalpis
o
4
b
c
,, pretoriensis
o
S
,, maeulatus
o
4
s
,, tbeobaldi
o
4
s
,, kartvari
o
,, willmori
o
4
s
Cbristopbersia kocbi .
o
4
s
Myzomyia culicijacies ...
o
4
s
,, listoni .
O
4
s
„ funesta .
o
4
s
,, turkbudi .
o
4
s
,, albirostris .
o
b
Pyretopborus cinereus .
o
4
s
,, nursei
o
4-
s
Pseudomyzomyia rossi .
o
4
s
,, ludlotvi
o
4 .
s
Pyretopborus jeyporensis
o
4-
b
,, costalis .
o
'{•
s
Cellia jacobi .
C)
4
s
75
belonging to the genera Mysomyia , Nyssorhynchus , etc., those of
the former being more triangular in shape and less scoup-like than
those of the latter. ( Vide Plate V, figs, i and 2.)
V. VARIATION AS DISPLAYED BY SCALES
A detailed account of scale structure would be foreign to the
purpose of this paper. But it is necessary in connection with other
variable features to summarise briefly the main facts regarding
scale variations in Anophelines.
This is the more necessary in that it seems to be taken for
granted that the only possible classification of the Anophelinae
based cm scales must be that actually employed by Theobald.
That this is a very limited view to take must be granted by all who
have paid close attention to scale characters. It may be found
that other scale characters not perhaps so conspicuous as those so
far employed may be of much greater use than has been suspected.
It will be useful therefore, even if the subject is only rapidly passed
in review, to indicate the broader principles of scale structure
variation. In doing so I shall take the opportunity of making
certain revisions in regard to the position certain species now
occupy, positions which it would be unfair later on to consider as
showing a discrepancy between the results of scale character and
colour marking classifications.
Head Scales , Marked variations in the head scaling are
restricted to a very few groups. The occurrence of linear head
scales has been used by James 1 to differentiate the genus Neoste -
thopheles (type N. aitkeni , James). This genus is clearly the same
as Theobald’s Stethomyia (type S. nitnba , Theo.), in which the
head scales are also linear. An examination of specimens of
A. corethroides , Theo., in the British Museum shows that this
unspotted winged species from Australia has linear head scales,
and should come in the genus Stethomyia and not in Anopheles
(vide also palpal characters). A . immaculatus> Theo., and
A. smithii , Theo., have head scales of the ordinary expanding
type.
Scaling of the palps . In some species the scales over most of
the palps are of small size and relatively appressed. In such
76
species the palp has a smooth thin appearance. In others the
scales are long and outstanding, giving to the palp a more or less
shaggy look. Difference in this respect, though not used in the
definition of generic groups, is by no means unimportant. One
would view with suspicion any supposed Cellia having smooth thin
palps, or a supposed Myzomyia with shaggy palps. Pm. rossi,
Giles, a species with shaggy palps until recently termed a
Myzomyia , is now known not to conform in scale characters with the
definition of this genus; presumably the related species
M. indepnata, Ludlow, M. ludlowi, Theo., M. mangyana, Banks,
are also rightly excluded. The species M. lutzi, Theo., at present
the only South American representative of Myzomyia , has shaggy
palps with a quite different type of ornamentation to that seen in
any other Myzomyia , and the correctness of the present position of
this species must be considered as problematical.
Between what may be called the suferpictus group of Pyre-
tophorus, which has very long thin palps, and P. costalis, Loew.,
there is also a very marked difference in this respect, which is
accompanied by marked structural as well as important scale
distinctions.
Antennal Scales. One of the characters of the genus
Calvertina ( C. lineata, Ludlow) is the presence of outstanding
scales on the second segment of the antenna. I have not seen a
specimen, but otherwise the scale characters from the description
seem very like those of Nyssorhynchus. The markings also read
very like those of N. fuliginosus, James and Liston.
Prothoracic Lobes. James (i) has called attention to the
importance of the patagia or prothoracic lobes from the point of
view of classification, and his separation of Patagiamyia (type
Pt. gigas, Giles) from Anopheles is based on the presence in the
former of a well-marked tuft of outstanding scales on these organs.
Using this feature one can distinguish as Patagiamyias, in
addition to those noted as belonging to the genus by James,
A. punctipennis, Say., and A. smitkii, Theo., both of which have
prothoracic tufts.
Judging from the light thrown by colour markings, a tuft of
scales on the prothorax is an extremely important character.
Unfortunately it is only in a proportion of species that the
77
description given notes the presence or absence of this character.
A prothoracic tuft appears to be of general occurrence in Myzo -
rhynchus ( barbirostris , umbrosus , sinensis , mauritianus ), there is a
tuft in Cycloleppteron grabhamii , in Arribalzagia maculipes and in
Myzorhynchella nigra. The tuft characteristic of Patagiamyia is
present also in Lophoschelomyia asiatica , Leicester. Myzorhynchus
wellingtonianus , Alcock, and Feltinella pallid opalpi , Theo., all of
which species would appear from their colour markings to be
related to P/. lindesayi , Giles. A tuft is present in Christophersia
kochi = (Cellia kochi, Donitz.), Neomyzomyia elegans , James,
CV//&Z squamosa , Theo., and curiously enough in P. costalis ,
Loew. On the other hand, a large series of species do not show
this structure.
At present, information is too scanty to enable any hard and
fast use to be made of the prothoracic scaling in classification, but
the table accompanying this section indicates approximately and
very tentatively the possible significance of the occurrence of a tuft
in this situation.
Mesothoracic Scaling. The presence or absence of broad scales
on the thorax has been used along with other characters to
differentiate a number of genera. Restricting observations to
certain groups, the presence or absence of broad mesothoracic scales
seems to serve the purpose of defining genera fairly well. By its
aid Afysomyia is fairly clearly marked off from Pyretophorus , etc.,
and if the shape and character of the scales is also taken into
account, this latter genus can be quite well differentiated from
Nyssorhynchus .
Extending our observations, a good many reasons for restricting
the significance of thoracic scaling become apparent, and any
groups in which primary divisions were formed on this character
would be very heterogeneous.
The great difficulty in scale structure is to know what
significance can be attached to particular variations. Thus
in comparing Pm. ludlowi, Theo., and P. costalis , Loew., which
differ very slightly in markings, character of the palps, etc.,
one does not know whether the presence in the latter of mesothoracic
scaling is to be taken as sufficient proof that they are unrelated or
not. The same difficulty occurs in the case of abdominal scaling
Table shewing tentative grouping on general scale characters.
78
79
in the species Ne. willmori , James, and N. maculatus , Theo.,
which it is practically impossible or very difficult indeed to
distinguish by any other characters than that one has a very much
more scaly abdomen than the other.
Metathoracic Scaling. Limited observations only, which have not
shown much promise, have been made in this respect. It was thought
that since the metathorax represented a primary division of the
thorax such scaling as was present might have more significance
than its mere extent or conspicuousness might at first suggest.
The halteres which spring from the metathorax, as one might
expect from their representing an undeveloped pair of wings, carry
scales. Usually the halteres are quite ball like, but in certain
species (e.g., Arribalzagia maculipes ) the wing-like character is
much more apparent (Plate VIII, fig. 39).
Wing Scales . Considerable differences occur in the shape of
the wing scales. The most noticeable variation is in the breadth
of the scales. There is also a difference pointed out by James, in
that whilst in some species the scales are broadest in the middle
(elliptical), in others they are broadest towards the free end
(oblanceolate). This character serves, according to James, to
distinguish between the genera Anopheles and Myzomyia .
Leg Scales . The genus Lophoschelomyia has been formed for
L. asiatica , Leicester, which has outstanding scales on the
femur. Recently another species showing the same appearance,
My. wellingtonianus , Alcock, has been described. Examination of
specimens in the British Museum shows that both these species are
not only probably related to one another, but have affinities with
Pt. lindesayi, Giles. Pi. lindesayi is at once differentiated by
possessing a very striking white band about the middle of the
femur. So far as one could see, this colour peculiarity was a pure
idiosyncrasy of the species. But the presence of a white band in
connection with the structural peculiarity of a prominent scale tuft
on the femur in L. asiatica and My. wellingtonianus suggests that
the band in lindesayi was once also accompanied by scale tufts.
The band is not quite in the same position in L. asiatica as in
My. wellingtonianus but its occurrence along with a scale tuft is
unlikely to be a mere coincidence. The three species in question in
any case appear to be closely related, and if the genus Lopho -
8o
schelomyia is retained, it should probably include Pt. lindesayi
(vide diagrams of wing markings, PI. VIII, figs. I, 3).
Abdominal Scales. Abdominal scaling, as pointed out by
James, shows two distinct characters depending upon whether the
scales project to form tufts or not.
Tufts are usually, but not always, associated with ordinary
scaling. They may be lateral or ventral. The genus Cellia , as is
well known, shows a marked development of lateral tufts; the same
condition is shown in Arribalzagia maculifes , Theo. The peculiar
species Ch. kochi , Donitz., shows a marked development of ventral
tufts, whilst the (apparently) totally unrelated Myzorhynchus
group has one such tuft (not, however, present in all species).
Pyretophorus is defined as having no scales on the abdomen.
On close inspection I find that there are a few scales in P . costalis ,
Loew., especially in the male, the appearances being much like
those seen in Pm. rossi , Giles. P. costalis , Loew., is the type of
the genus, so that the definition of Pyretophorus (abdomen without
scales) now held will not serve. The group of mosquitos like
P. nursei seem to have a quite scaleless abdomen.
A common condition is that in which the last two or three
segments only carry scales. This is best known in Nyssorhynckus ,
but it also occurs in Lophoschelomyia , Manguinosia , and Kerteszia.
In Cellia , Neocellia , Christ ophersia, and Arribalzagia the
abdomen carries large numbers of scales.
In the table I give a quite tentative and provisional grouping
based on scale structure, adopting as broad an outlook as possible.
It will be noticed that I have given great prominence to the
condition of the prothoracic lobes. It is possible that this may
require modification, but at present all my observations have gone
to show the extreme significance of the presence or absence of a tuft
of scales on the prothorax.
VI. CLASSIFICATION, PHYLOGENY AND GEOGRAPHICAL
DISTRIBUTION OF THE ANOPHEUNAE
A tabular statement of groups as characterised, and to some
extent defined, by structural characters and colour markings
accompanies this paper. Though scale characters are given in this,
they only enter very occasionally into the formation of the groups,
and are purposely placed in parenthesis in order that their close
agreement with the general scheme of classification given may be
apparent. It will be seen that in the main Theobald’s genera stand
out clearly. Considerati6n of other characters, however, modifies
Theobald's divisions as follows: —
(1) It shows certain genera to be composite.
(2) It groups the genera together so that one has some realisation
of their zoological value.
(3) It shows that it is not impossible that a system of quite
good genera, more or less on Theobald’s line, could be
with advantage retained, and that it is premature to
resort as yet to the abolition of this grouping.
I do not propose, in the present paper at least, to offer any new
system of nomenclature, which would require, in order not to add to
to the confusion already too apparent, the most careful and detailed
systematic study of the subfamily. It will be sufficient for the
present if the quite natural character of Theobald’s genera, taken as
a whole, is emphasized. What seems mostly now required is
accurate and detailed descriptions of species, with a really
comprehensive and authoritative revision of the Anophelinae. But
a list of the groups as defined by structural characters and colour
markings, and a tentative arrangement of these as they appear to
be phylogenetically related, should not be open to any objection.
Retaining Theobald's nomenclature, but noting the necessity of a
number of emendations as regards the species at present contained
in these genera, and subdividing the genera where necessary, the
following represents, as far as one can judge, the phylogenetic
relationship of these groups.
Tentative Phylogenetic Arrangement of the Anophelinae
82
Towards elaboration of ornamentation and scale development
l
Stethomyia
«3
In this arrangement, phylogenetic advance is associated with
both an increase in colour markings (i.e., the development of white)
and in scaling. It is not difficult to arrange a correlation table in
which increasing degrees of whitening and increased degrees of
scaling form the ordinates and abscissae respectively. The striking
nature of the correlation between the two series of characters is then
made very apparent by the grouping of species about the diagonal
line. The same correlation is also very evident in the phylogenetic
scheme given.
Considering the fact that the Culicinae are all scaly, one might
think it probable that Anophelines were a branch in which scales
were disappearing. But my own observations on this point make
it more probable that it is the other way. The more scaly an
Anopheline is the more advanced phylogenetically it would appear
to be.
Certain of the groups, it will be seen, occupy a much more
primitive position in the scheme than others, and as this seems to be
demonstrated by whatever character they are considered, and as
there are definite characters by which one can distinguish these
apparently old type species, I have named them Protoanopheles.
The peculiar Australasian type I have called Neoanopheles. The
ordinary Anophelines exhibiting the regular colour scheme I have
outlined may approximately be termed the Deuteroanopheles. The
nature of these groups, and the characters on which they are based,
can be obtained from the large table accompanying this paper.
Briefly stated, the constitution of the main groups noted is as
follows: —
Protoanopheles .
Stethomyia.
Anopheles.
Patagiamyia and Lophoschelomyia.
Myzorhynchus, Cycloleppteron, Arribalzagia, etc.
Deuteroanopheles .
Mysomyia.
Pyretophorus.
Pseudomyzomyia, etc.
Nyssorkynchus.
Neocellia.
Cellia.
Branch A.
Branch B.
Branch C.
*4
Neoanopheles.
Many spotted winged species and their related types.
Between branches A and C of the Deuteroanopheles there are
distinct differences in ornamentation, such as, for example, a much
greater tendency to a completely broken third costal spot and
narrow costal interruptions, but the most striking difference is in
the much greater intensity of the black and white ornamentation.
The origin of the branch B is difficult to determine. It is especially
interesting as including some of the most dominant species,
i.e., Pm. rossi, P. costalis, Ne. stephensi, Pm. ludlowi.
Geographical Distribution
Having on structural grounds arrived at certain groupings, it is
interesting to note the geographical distribution of these groups.
Stethomyia, though it contains very few species, has a
distribution area which includes South America and Australia, as
well as India and Malay.
Myzorhynchus , if we consider the closely-related genus Cyclo-
leppteron, also has a very extended distribution, which includes
South America and Australia.
Anopheles and Patagiamyia are the dominant genera of Europe
and North Amerca. There is, however, what appears to be an
Australian representative of this branch, as well as hill species in
the East.
In the case of the Deuteroanopheles, certain of the genera have
a much more restricted distribution. Thus Pyretophorus, in the
restricted sense in which I have been using the term, is dominant
in Africa (North), occurs in South Europe and in the extreme
North-west of India. It does not occur in South America or
Australia, and it is scarcely represented in the Anopheline fauna of
India and Malay.
Myzomyia is essentially African, Indian and Malayan.
Nyssorhynchus is also African and Malayan, with a distinct
dominance in the latter area and in India.
The most widely distributed of the genera of the Deutero-
85
anopheles is Cellia , and this fact is not out of keeping with the
position assigned to this genus in the scheme of phylogeny.
The Neoanopheles are distinctly Australasian and Malayan.
More than the above outline of the main features of geographical
distribution of species cannot be attempted here, but it will be seen
that to a large degree consideration of the geographical distribution
of species gives distinct support to the ideas respecting phylogeny
which I have put forward in this paper.
VII. GENERAL CONCLUSIONS
Colour markings, equally with structural characters, can be utilised
in natural classification of the Anophelinae and in the placing
of species in groups to which they have affinities. A classification
based on colour marking, supported by structural differences in the
palps and larval characters, approximates very closely in regard to
the groups formed with one based on scale structure. It shows,
however, the relation and affinities between the groups much more
clearly than does scale structure classification as at present employed
in the distinction of genera.
Colour markings, and general characters as a whole, seem to
point to there being a group of more primitive forms (PROTO-
anopheles) occurring as old world species (Anopheles, Myzo-
rhynchus and Patagiamyia ), and as new world representatives of
this group Cycloleppteron , Arribalzagia and Myzorhynchella. There
is also a distinct group corresponding to Donitz’s Australasian
species with more than three spots on the sixth vein (NEOanopheles).
Their area of special prevalence is Australasia and Malaya.
The majority of Anophelines belong to a group the colour
markings of which are all on a given colour scheme (DEUTERO-
ANOPHELES). This group shows two main divisions, characterised
by a notable difference in the intensity of their coloration and the
‘ effectiveness * of ornamentation, possibly arising from two different
lines of evolution. Their area of special prevalence is an area
including Africa, South Asia and Malay.
I am very greatly indebted to Prof. R. Newstead and to
Mr. H. F. Carter for their great kindness in furnishing me with
86
material for studying a number of African and South American
species, including specimens of Cy. grabhamii, without which I
could not have carried out many of my observations. I am also
indebted to Prof. Newstead and Mr. Carter, as well as to
Mr. F. W. Edwards, for their kindness in giving me every facility
to make use of the collections at Liverpool and the British Museum
respectively.
I also wish to acknowledge my indebtedness to Prof. G. H. F.
Nuttall and to Mr. H. Scott, at Cambridge, for help in regard to
literature and in other respects.
»7
VIII. TABLE SHEWING GROUPING OF SPECIES OF ANOPHELINAE
ACCORDING TO COLOUR MARKINGS AND OTHER VARIABLE
CHARACTERS
A. NODAL CENTRES AT CROSS VEINS AND BIFURCATIONS
DARK. (PROTOANOPHELES.)
A'. Costa devoid of any light areas even at apex. Wings
USUALLY WITHOUT ANY PALE MARKINGS.
1. Palpi markedly heterodactylous with marked relative elongation of
the second segment. Attitude culex-MVc. Wings entirely
unspotted. Palps unbanded. Legs entirely without markings
even knee spots being absent. Small mosquitos.
(Larval antenna without branched hair. Palmate leaflets lanceo¬
late.)
Head scales linear. No scales on prothorax, mesothorax or abdomen.
Corresponds exactly with Stelbomyia , Theo.; Neostetbopbeles.
James.
5 . nimba y Theo.
S. aitkeniy James and Liston.
S. culiciformtSy James and Liston.
5 . coretbroidesy Theo.
Probably synonyms— S. treacberi , Leicester ; S. fragilis ,
Theo.; 5. pallida , Ludlow.
2. Palps orthodactylous. Attitude anopbeles-WVt. Palps thin,
unbanded. Primitive spotting may be present. No admixture
of light and dark scales. Knee spots usually present otherwise
legs are quite unornamented.
(Larval antenna with branched hair (small), palm-leaflets lanceolate
and serrated).
Head scales expanded. No prothoracic tuft. No scales on
mesothorax or abdomen.
Corresponds with Anopheles sense of James.
A. maculipennisy Mcigen.
A. bifureatuSy Linn.
A . nigripeSy Staeger.
A. algerienstSy Theo.
A. barianensisy James.
A. barberiy Coq.
A. immaculatusy James. (?).
A. eiseniy Coq. <?).
A. crucians , Wied. (?).
88
A". Costa with at least one pale interruption which may
BE AT THE APEX.
(b') Wings without prominent admixture of pale and dark
scales .
3. Cotta dark broken oply at apex or at tubcoita with or without
•mall basal interruption. Palpi usually unbanded. Primitive
•potting prominent. Tartui unbanded.
(Larva with branched hair on antenna. Palmate leaflets with
filament but showing approach to lanceolate type.)
Head scales expanded. Tuft on prothorax. No broad scales on
mesothorax. May or may not be scales on last segments of
abdomen.
3«. With white band on femur which may be associated
with tufts of outstanding scales in this situa¬
tion.
Lopboscbelomyia.
L. cuiatica, Leicester.
L. linietayi , Giles.
L. wcllingtonianus, , Ale ode.
3 b. Without white band or femur.
L. (?) atratipes, Skuse.
4. Costa prominently spotted. (Usually two, not more than three,
main spots). Palps banded. Primitive spotting may be present
(but not conspicuous). White spots on wings for most part
represented on lower surface. Large mosquitos.
(Larval antenna carries a branched hair. Palmate leaflets with
filament.)
Head scales expanded. Tuft on prothorax. No scales on meso-
thorax (or abdomen).
Corresponds to Patagiamyia , James.
Pt. gigas , Giles.
Pt. lindens is, James.
Pt. punctifennis , Say.
Pt. mitbi , Theo. (Unspotted wings.)
Probably
A. for mas us , Ludlow.
Perhaps
A. franciscanus , McCracken.
A. perplex am, Ludlow.
A. pseudopunctipennis , Theo.
Would probably include also
FeltineUa pallidopalpi, Theo.
5. Costa dark with narrow interruptions but with four main costal
•pots. Wings markedly dark beneath. Palps shaggy.
(Larval antenna without branched hair. Palmate leaflets lanceolate.)
Head scales flattish. Broad scales on meso thorax. No scales on
abdomen.
Myzorbyncbella.
8 9
(£") Wings with prominent admixture of dark scales. Pale
spiffs on the wing deficient as a rule on under surface.
Hence , when third spot is light it shows dark border
of projecting dark scales seen through wing membrane.
Primitive spotting the rule. Sixth vein with at
least one scale cluster of dark scales.
{Larval antennae with branched hair. Palmate leaflets
lanceolate and serrated. Frontal hairs markedly
branched.)
Prothoracic lobes with tuft.
6. Legs not speckled. Costal edge dark with two (at most three)
minute pale interruptions. Basal portion of costa unbroken
by pale areas. Palps markedly orthodactylous, shaggy, un¬
handed or with scheme A ornamentation. Large mosquitos
Old World species.
No broad scales on mesothorax. Some ordinary scales present
or absent on abdomen. A ventral tuft of dark scales usually
present on penultimate segment of abdomen.
Corresponds to Myzorbyncbus.
My. bar biros tr is, Van der Wulp.
My. pseudobarbirostris, Ludlow.
My. bancroftii , Giles.
. My. umbrosus , Theo.
My. stracbani , Theo.
My. sinensis , Wied.
My. pseudopictus , Grassi.
My. paludisy Theo.
My. mauritianus , Grandprc.
7. Legs markedly speckled. Costal edge may be dark as in last group
or broken into distinct spots. Accessory spot often present.
Palps shaggy with scheme A ornamentation. Large mosquitos.
New World species.
2 a. Sixth vein with two dark spots. Wing with inflated
scales but resembling that of Myzorhynchus
in general arrangement.
No broad scales on mesothorax.
CycloUppteron
CyclaUppteron grabbamii , Theo.
2b. Sixth vein with many spots.
Broad scales on mesothorax. Abdomen with many
scales and lateral tufts.
Arribalzagia.
Ar. maadipes , Theo.
Cy. mediopunctatus , Theo.
{Ar. pseudomaadipes).
{Ar. malefactor).
{Cy. intermedium).
90
B. NODAL CENTRES AT CROSS VEINS AND BIFURCATIONS
PALE. COSTA WITH FOUR DISTINCTLY MARKED
MAIN COSTAL SPOTS.
B'. Palps Ornamented on Scheme A—C. Costa without
Accessory Spots other than the basal ones. Not
MORE THAN THREE DARK SPOTS ON SlXTH LONGITUDINAL
Vein. (DEUTERO ANOPHELES.)
(a) Tips of hind tarsi not white.
a!. Palps markedly heterodactylous. Tips of hind tarsi never
white. Banding of front tarsal joints never broad .
Legs never markedly speckled. fVing markings of
species on fixed scheme {Scheme A). If three spots
are present on sixth vein there is a pale interruption
on upper branch of second longitudinal. Completely
broken third costal spot unusual.
{Larval antenna without branched hair. Palmate leaflets
with filament. Frontal hairs usually simple
unbranched.)
No prothoracic tuft. No scales on abdomen , even in male.
1. Sixth vein with two or 1 cm dark areas. No pale internodal spot
on branch of fifth. Branches of second long, rarely with pale
interruption. Still more rarely interruptions on branches
of fourth. Fifth vein dark at junction of branch.
Mesothorax without broad scales.
Corresponds to Myzomyia in restricted sense (i.e., not including
Pm. rossi , Af. lutzi , etc.).
Af. culicifacies, Giles.
Af. nUi , Theo.
Af. rbodesicnsis , Theo.
Af. bebcSy Donitz.
Af. umbrosa, Theo.
Af. listoniy Liston.
Af. funesta , Giles.
P. sergtntii , according to wing.
2. Sixth vein with three dark spots or two. Pale internodal spot
on branch of fifth. Branches of two and four veins with pale
interruptions. Fifth vein usually pale at junction of branch.
Palps peculiarly long and thin. Not infrequently show apical
dark band (four palpal bands) or apex dark.
Broad scales on Mesothorax.
Pyretapborus 1 (superptetus group).
P. super pic l us, Grassi.
P. nursei, Theo.
P. nigrijasciatusy Theo.
P. cleopatrae , Willcocks (MSS.)
P. cardamatisiy Newstead and Carter.
P. distinctly Newstead and Carter. Costa
unusual but wing otherwise showing typical
Pyretopborus condition.
P. paUstinensisy Theo.
P. cirureuSy Theo.
9 1
a”. Palp not markedly heterodactylous.
3. Upper branch of second vein with pale interruption.
Tip of hind tarsus not white. Palps destinctly orthodactylous but
with whole of apical segment pale. Banding of front tarsus
broad. Apical segment of palps in spite of length pale through¬
out. Hind tarsi not white but speckling usual. Whitening of
wing considerable with completely broken third costal spot
usual. All intemodal pale spots developed and some residual
pigment areas usually obliterated.
A composite group according to scale structure.
Prothoracic tuft. Preapical costal spot broken. Scales
on mesothorax. A few scales on last segment
of abdomen especially in male.
Pyretopborus II.
P. costal is, Loew.
(P. pseudocostalis , Theo. ?).
(P. merus y Donitz ?).
(P. marsbaUii, Theo. ?).
No prothoracic tuft. Preapical costal spot not broken.
A few broad scales on mesothorax. A few
scales on last abdominal segment especially
in male.
P seudomyzomyia, Theo. = Nyssomyzomyia, James.
Pnt. rossi, Giles.
Pm. indefinata , Ludlow.
Pm. ludlotoiy Theo.
No prothoracic tuft. Nyssorhynchus-like scales on thorax.
Ne. stepbensi.
4. Upper branch of second without pale interruption. Palps ortho¬
dactylous and with scheme A pattern. Bridging of pale spots
on costa by dark areas on first long vein marked feature.
Prothoracic tuft. Broad scales on mesothorax. Abdomen with
lateral tufts.
Cellia squamosa , Theo. Related to Myzorbyttcbus and
Cellia. A peculiar species.
Isolated position in this table due to fact it does not
possess white tips to tarsus. Note prothoracic
tuft.
(b) Tips of hind tarsi white. Ornamentation very black
and white. Speckling and banding usual.
Full development of internodal pale areas of
wing, but delayed appearance of pale area on upper
branch of second vein.
(Larval antenna without branched hair. Palmate
leaflets with filament. Frontal hairs often show
some degree of branching.)
92
Characteristic broad white creamy scales on meso-
thorax.
5. Palp* ordinarily three banded (scheme B). Orthodactylous or
with some tendency in light forms to be slightly heterodactylous
(N. maculatus).
Abdomen with scales on last few segments ( Nyssorbyncbus ) or on
many segments ( Neocellia ) but without lateral tufts. No
prothoracic tuft.
Dark group. Upper branch of second long vein without
interruption. Hind tarsi usually show un¬
interrupted white area extending over several
segments. Palps destinctly orthodactylous,
ornamented scheme B, with tendency in
individuals to revert to A.
N. fuliginosus , Giles.
N. nivipes , Theo.
( N . freerae , Banks.)
(N. pbilUpinetuiSy Ludlow.)
Ne. fowleri, Christophers.
N. jamesi, Theo.
N. pretoriensis, Theo.
N. maculipalpisy James and Liston.
Light group. Upper branch of second long vein
with interruption. Hind tarsi usually with
alternate black and white areas. Palps with
shorter terminal segment than last group
and with scheme C ornamentation (two
broad apical bands).
N. maculatus, Theo.
N. tbeobaldiy Giles.
Ne. tcillmoriy James.
6. Palps ordinarily four banded (scheme A) and markedly orthodac¬
tylous.
Abdomen with lateral tufts. Prothoracic tuft in some species.
Cdlia. Dark group. Bridging of pale spots on costa common.
Also absence of interruption on upper branch
second.
C. argyrotanisy Desvoidy.
C. albimandy Wied.
C. jacobiy Hill and Haydon.
C. cineta , Newstead and Carter.
(C. squamosa, Theo. Hind legs not white, vide
above.)
(C. squamosa , tw. arnoldi. Hind legs not white,
vide above.)
Light group. Upper branch of second interrupted.
Costal spots not bridged.
C. pulcberrima , Theo.
C. pharoensisy Theo.
93
B". Palps with Scheme D ornamentation. More than
THREE DARK SPOTS ON THE SlXTH VEIN. ACCESSORY
COSTAL SPOTS OFTEN PRESENT IN ADDITION TO BASAL ONES.
(NEOANOPHELES.)
With prothoracic tuft.
My. nataltiuit , Hill and Haydon. •
P. xcatsoniij Leicester. *
Nm. elegant, James. *
(Nm. UucospbyruSy Donitz.) *
N. annulipes, Walker. •
N . master t, Skuse. •
Cb. kocbty Donitz.
Query any prothoracic tuft.
M. punctulata, Donitz. •
N. deeeptor, Donitz. •
N. tborntoniiy Ludlow.*
N. kartoariy James and Liston.
All species show same character of palpal ornamentation.
• Shows more than three dark spots on sixth.
REFERENCES
1. Jams, S. P. (1910). A new arrangement of the Indian Anophelines. Records of the
Indian Museum, VoL IV, No. 5.
2. Mayx* (1897). On the color and color patterns of moths and butterflies. Proc. Bost.
Soc. Nat. Hist., Voi. XXVII, pp. 243-330.
3. Hill and Haydon (1907). A contribution to the study of the characteristics of larvae of
species of Anophelina in South Africa. Annals of the Natal Gov. Museum, Vol. I,
Part 2.
4. Donitz (1902, 1903). Beitrage zur Kenntnis der Anopheles. Zeit. f. Hygiene, Bd. XLI,
p. 15, and Bd. XLIII, p. 215.
5. Nuttall and Shiplzy (1901, 1902, 1903). The structure and biology of Anopheles.
Jour, of Hygiene, VoL I, Nos. 1, 2 and 4, pp. 45, 269, 451 j Vol. II, No. 1, p. 58 ;
Vol. Ill, No. 2, p. 166.
6 . Edwazds, F. W. (1911). Some new West African species of Anopheles (sensu lato),
with notes on nomenclature. Bull. Ent. Res., Vol, II, Part 2, p. 141.
94
Fig. i.
Fig. 2.
F>g- 3 -
Fig. 4.
Fig- 5 -
Fig. /•
Fig. 8.
Fig- 9 -
Fig. 10.
Fig. 11.
Fig. 12.
Fig- 13 -
Fig. 14.
Fig. 15.
Fig. 16.
Fig. 17.
EXPLANATION OF PLATES
Plate V
Nymphal trumpets of A. bifurcatus , Linn.
Nymphal trumpet of Ne . willmori l James.
Leaflet of palmate hair of My. sinensis , Wied.
Leaflets of palmate hairs. (d) C. squamosa var.
arnoldi, after Newstead and Carter, (b) N. annulipes ,
Walker, after Theobald. ( c) A. bifur catus. (d) Cyclo -
leppteron grabhamii y Theo., after Theobald, (e) Myzo-
rhynchus (?) natalensis , Hill and Haydon, after these
authors. (/) P. ardensis , Theo., after Hill and Haydon.
Antenna of larva, (a) A. bifur catus. ( b ) A. puncti -
pennis , Say., after Smith.
Female palp of Afy. barbirostris , Van der Wulp, showing
orthodactylous character and unbanded condition.
Female palp of C. pulcherrima , Theo. Orthodactylous
with scheme A ornamentation.
Female palp of lindesayi , Giles. Orthodactylous and
unbanded.
Female palp of P. nursei , Theo. Heterodactylous with
appressed scales and scheme A ornamentation.
Female palp of P. jeyporensis , James. Heterodactylous
with appressed scales and ordinary Deuteroanopheles
type of ornamentation, i.e., scheme B, with involvement
of the whole of the apical segment.
Female palp of P . costalis , Loew. Orthodactylous with
scheme B ornamentation.
Female palp of M. funesta , Giles. Heterodactylous with
appressed scales and scheme B ornamentation.
Female palp of N. maculatus y Theo. Scheme C ornamenta¬
tion.
Female palp of N. maculatus contrasted with palps
ornamented on scheme D. {a) N. maculatus. (o) M.
punctulata , Donitz. (c) Ch. kochi , Donitz. ( d ) N.
karwari t James. After James.
Sketch of palp of S. corethroides y Theo., showing remark¬
able modification in the relative lengths of the different
segments. Heterodactylous. Type 2.
Palp of S. nimba , Theo., after Theobald.
96
Plate VI
Fig. i. Diagram showing structural features of wing and leuco-
genetic centres. Nodal points are shown by small
circles, intemodal by larger ones. Pigment centres are
shown dark.
Pigment centres.
AcAc n . Basal accessory pigment areas.
i.m.c. Inner main costal spot centre.
m.m.c. Middle main costal spot centre.
s.a.m.c. Subapical main costal spot centre.
a.m.c. Apical main costal spot centre.
2 r. t 2 a. Basal and apical centres of stem of second
vein.
2 r b. t 2'a. Inner and outer centres of upper branch of
second longitudinal vein.
2 2 n b ., 2 n c. Outer, inner and middle centres of
lower branch of second vein.
3 a. y 3 b. Residual pigment centres of third longi¬
tudinal.
4 r., 4 a. Centres on stem of fourth longitudinal.
4#., 4 'b . 9 4 n a. y 4 ”b. Centres on fork of fourth longi¬
tudinal.
5V7., 5 'b., 5 f c. Centres on branch of fifth longitudinal.
5r., 5"<7. Characteristic centres of fifth longitudinal
vein.
6 r t 6 m , 6 a. Centres for sixth vein. 6 r. is Donitz's root
spot of sixth.
C2» ^a» C 5 . Nodal
Nodal points. points in connection with
h . Humeral. cross veins.
Sc. Subcostal. C.i f . (?) Nodal point con-
Ap. Apical. cerned in the formation of
S. Sector. the inner pale spot on costa.
5 '. Accessory sector. C. i*. (?) Nodal point con¬
cerned with subapical spot.
i, 2', 2", 3, 4', 4*, 5', 5", 6. Nodal points on wing
margin.
7c S 7 CyclolefifiCtron.
3l<§.S. 7*a amy ca
Si^.1%.. h<fSsorh.ynttr.us j CtULX tic
, I $ * iea(e
° = . °_ ■ ' 3, dmkica(«d. U^catfc.. /tig./ 3 . htotny2.omyt.a..
O » (nlirnodal 0 .«,,«< d M »t aMd.m*(r£cA(c
^ r (UULlux^
C. Tinting fr* Co /w^.
97
Internodal points.
2 1 , 2 2 , 2 3 . Internodal points on branches of second vein.
The relatively late appearance of 2 1 is
characteristic of Nyssorhynchus and dark
Cellias.
3, 3. One or two internodal points of third vein.
4, 4 1 , 4 2 . Internodal points of stem and branches of
fourth vein.
5 1 , 5 2 » 5 3 - Points on main vein and branch of fifth vein.
6 l , 6 2 . Points on sixth vein.
Fig. 2. Portion of the wing of a species of tipulidae , showing
venation homologies and extra cross veins.
h. Humeral cross vein.
C. 1'. Cross vein joining subcosta to first longitudinal.
C. 1". Ditto upper branch of second to
first longitudinal.
R lt R 2 , R 39 /? 4 + 8 , Radial system.
R. S. Radio-sector.
5 , As in figure 1.
S'. Ditto.
Fig. 3. Showing area of cross veins, and relation to nodal point S.
Cv 2% Cv 3 , Cv 4 , Cv s . Cross veins 2-5.
S. = Sector, S'. = Accessory sector nodal point.
sc. = Subcosta.
Fig. 4. Wing of a species of Chironomus , showing pale spots on
wing membrane in^lving the hair-like scales covering
the wing and aggregation of hair-like scales to form
dark spots at the area of the cross veins.
Fig. 5 to Fig. 13. Diagrams of representative wings, showing
arrangement of nodal and internodal points as developed
in different groups.
Dark spots represent nodal points which remain
pigment areas. Circles half dark represent pale spots
not developed on under surface of the wing. Small
black dots in circles mean admixture of dark and light
scales present. Dark stars mean scale aggregations.
98
Plate VII
Fig. i. Wing of A. maculipennts , showing primitive spotting.
Fig. 2. Wing of My . umbrosus , showing primitive spotting, want
of representation of pale spots on under surface of wing,
and admixture of dark and light scales. A scale cluster
on the sixth vein is also shown.
2a. Under surface.
Fig. 3- Wing of M. funesta var., showing first appearance of
nodal points with few internodal centres. Pale scaling
not intensely white.
Fig. 4. Wing of N . maculatus , showing extreme development of
nodal and internodal centres, with residual pigment
areas well displayed. White scaling intensely white.
too
Plate VIII
Diagrams of the spotting of the wings in a number of species of
Anophelinae, especially to show arrangement of costal spots.
Figs. 1-38.
Fig. 39. Halter of Cy. mediopunctata , showing resemblance to
unexpanded wing, with light scaling above and
intensely dark scaling beneath.
1. Patagiamyia lindesayi % Giles.
2. My . wellingtonianuSy Alcock.
3. Lophoschelomyia asiatica , Leicester.
4. Patagiamyia gigas , Giles.
5. Patagiamyia simlensis , James, var.
6. Myzorhyrtchus umbrosus , Theo.
7. Ditto under surface.
8. Myzorhyrtchus mauritianus , Grandpre.
9. Myzorhynchus*sinensis , Wied.
10. Cycloleppteron grabhamit , Theo.
11. Ditto under surface.
12. Cycloleppteron mediopunctatus , Theo.
13. Arribalzagia maculipes , Theo.
14. Myzorhynchclla nigra , Theo.
15. Ditto under surface.
16. Cellia squamosa , Theo.
17. Cellia squamosa , Theo.
18. N yssorhynchus fuliginosus , Giles.
19. Myzorhyrtchus (?) natalensis , Hill and Haydon.
20. Pyretophorus (?) watsonii , Leicester.
21. Neomysomyia leucosphyrus> Donitz.
22. Myzomyia (?) punctulata , Donitz.
23. C hristophersia hoc hi , Donitz.
24. Nyssorhynchus (?) annulipes , Walk. (Costa only
shown.)
25. Pyretophorus costalis , Loew.
26. Cellia jacobi , Hill and Haydon.
27. Myzomyia culicifacies , Giles.
28. Myzomyia listoni , Liston.
29. Pyretophorus cinereus , Theo.
30. Pyretophorus nursei , Theo.
31. Pyretophorus cardamitisi , Newstead and Carter.
32. Pyretophorus chaudoyei 9 Theo.
33. Pyretophorus jeyporensis , James.
34. Id yssorhynchus theobaldi , Giles.
35. Neocellia stephensi , Liston.
36. Pseudomyzomyia rossi , Giles.
37. Pyretophorus ardensis , Theo.
38. Pyretophorus (?) atratipes t Skuse.
C. Tittling Co.. Ltd.. Imp.
101
A STUDY OF THE
POSTERIOR NUCLEAR FORMS OF
TRYPANOSOMA RHODESIENSE
(Stephens and Fantham) IN RATS
BY
B. BLACKLOCK, M.D., D.P.H.
From the Runcorn Research Laboratories
(Received for publication 5 February , 1913)
Trypanosomes, obtained from a human source, presenting a
posterior position of the macronucleus were first described by
Stephens and Fantham. 1 Upon this morphological peculiarity they
founded their species Trypanosoma rhodesiense. Since the publica¬
tion of their paper, a similar condition has been described in
trypanosomes from human sources by various observers in different
regions. The patient whose strain they described contracted the
disease in a portion of Rhodesia in which the absence of Glossina
palpalis rendered it necessary to seek a different carrier. Stannus
and Yorke 2 discovered posterior nuclear forms in a strain taken
from a case of human trypanosomiasis in Nyasaland. More
recently Stannus 3 has noted the presence of these forms in a strain
obtained from a case of human trypanosomiasis from Portuguese
East Africa, while the Royal Society's Commission 4 confirms the
observation of Stannus and Yorke with regard to the presence of
posterior nuclear forms in strains from human trypanosomiasis in
Nyasaland. The Commission bears out their conclusion that the
trypanosome of the human trypanosome disease of Nyasaland is
probably identical with T. rhodesiense .
The occurrence of posterior nuclear forms is not, however,
confined to strains of human trypanosomiasis. Such forms have
also been recorded among animal trypanosomiases, for example in
T. pecaudi by Wenyon, 6 in a strain of T. equiperdum by Yorke
and Blacklock, 6 and in a strain of T. brucei from Uganda by
Blacklock. 7
102
THE INCIDENCE OF POSTERIOR NUCLEAR FORMS IN
T. RHODES1ENSE
Stephens and Fantham (loc. cit.) observed that the forms of
trypanosome which presented a posterior position of the nucleus
were the short and stumpy ones, and that in rats forms having this
peculiarity made their appearance in the peripheral blood about the
fifth or sixth day of the disease, and increased in number to the
seventh or eleventh day, when they formed about 6 % of the
parasites present.
SHORT AND STUMPY
The term ‘ short and stumpy ’ appears to be used in somewhat
different senses by different observers, in relation to trypanosomes.
For example, Bruce (loc. cit.) says ‘ There is no free flagellum in
the short and stumpy forms.’ In a table attached, however, he
shows that the amount of free flagellum in the ‘short and stumpy’
forms (620 individuals) averages 0*6. Stephens and Fantham
(loc. cit.) say of the stumpy forms of T. rhodesiense which have a
posterior nucleus that there is a well-marked blepharoplast and
a very short free flagellum. It appears, therefore, to be not quite
decided what constitutes a ‘ short and stumpy ’ form. Is it a
trypanosome which measures less than a given number of microns,
e.g., 21; is it a trypanosome which has absolutely no free
flagellum; or is it a trypanosome which is not only less than a
certain length, but also has absolutely no free flagellum? In the
experiments given below the large majority of forms which had a
posterior nucleus measured less than 21/u, but numbers were found
which exceeded this length. Again, of the parasites which
conformed to the definition as regards shortness, numbers had
absolutely no free flagellum, while numbers had a very definite
portion of flagellum free. In some cases this portion was of
considerable length. It was not impossible, for example, to
discover forms which while they measured only 19 /i in length, yet
possessed a free portion of flagellum amounting to 4 /t.
One finds thus that when studying the posterior nuclear forms
in these experiments, one is not dealing with short forms in the
strict sense that they measure in all cases less than 21 p nor with
>°3
stumpy forms in the sense that they possess absolutely no free
flagellum. There are overlapping forms which do no come under
both categories, in fact there are a few which come under neither,
yet present the posterior nucleus. For practical purposes, however,
it may be said that the posterior nuclear forms, whether free-
flagellated or aflagellar, which exceed the measurement of ‘short
forms,’ were few in number. The experiments detailed below were
undertaken in order to determine in a given infection in rats the time
of appearance of posterior nuclear forms in the peripheral blood,
and the numerical relationship which these forms bear to other
forms of trypanosome present from day to day.
METHOD OF CARRYING OUT THE EXPERIMENTS
A guinea-pig infected with T. rhodesiense and having numerous
parasites in its blood was utilized as the source of infective material.
Four groups (A, B, C, D) of rats, each group containing six
rats, were selected, the rats being as nearly as possible of the same
weight. The parasites per cubic millimetre in the blood of the
guinea-pig were estimated by means of the Thoma Zeiss haemo-
cytometer, and dilutions made in a warmed mixture of I % sodium
citrate and 0'85 % sodium chloride. The dilutions were made so
that the rats of the first two groups received 1,000,000 trypano¬
somes, the rats of the third group 4,000,000, and the rats of the
fourth group 8,000,000. The animals were all inoculated intra-
peritoneally, the amount of the injection in each case being o'5 c.c.
INCUBATION AND DURATION
The average incubation period of Group A (1,000,000 trypano¬
somes) was 4 8 days, the average duration of the disease 13 6 days.
Group D (8,000,000 trypanosomes) had an average incubation
period of 4'5 days and an average duration of 15*5 days. Table I
gives details as to the incubation and duration of the disease in the
individuals of each group. From this table it appears that no
definite variation in either incubation or duration could be
attributed to the relative numbers of trypanosomes injected.
io 4
Table I.—Giving incubation and duration of infection with T. rbodesunse in 24 rats.
Group
No. of Experiment
Incubation in days
Duration
A
Rat 2231 A .
5
*3
1,000,000
n B
4
>3
Trypanosomes
« c .
5
f 4
inoculated
„ D .
6
14
" E .
4
>4
„ F .
5 -
>4
B
Rat 2232 A .
5
13
1,000,000
„ B .
6
*3
„ C .
5
16
U D .
5
16
„ E .
4
*3
.. F .
6
*3
C
Rat 2229 A .
5
»3
4,000,000
,, B .
4
12
c .
4
11
;
D .
4
10
1
E . !
4
13
„ F .
6
*3
D
| Rat 2230 A .
4 |
<7
;
8,000,000
i „ B .
4
>7
„ c .
4
15
i „ D .
5
*4
„ E .
5
15
! ,, F .
5
1 *5
Average ...
l
47
137
ENUMERATIONS
From the day on which parasites first appeared in the peripheral
blood until the death of the animal, a film was examined daily in
the fresh state, and a thin film dried and stained. The dried films
were fixed in absolute alcohol and stained with IGiemsa’s stain.
i. Preliminary count. In each stained film a count of 200
parasites was made wherever possible. Every trypanosome
met with was counted, whether long, intermediate or short,
dividing or non-dividing. The number of posterior nuclears was
noted, non-dividing forms only being chosen. The result, therefore,
gave the number of non-dividing posterior nuclear forms per
200 of all forms. The posterior nuclear forms were classified
carefully according to the position of the nucleus. Those
forms were classified as A in which the nucleus, although
definitely posterior to the centre of the parasite (excluding free
flagellum) still lay close to the centre. C forms were those in which
the nucleus lay adjacent to the blepharoplast, while B forms were
intermediate in position. The drawings (Text-fig. 1) give the
positions indicated for each group.
f §
B C
Tixt-Fig. 1. Showing various position* of nucleus. A. B. C.
RESULT OF THE PRELIMINARY COUNT
The first trypanosomes which appeared in the peripheral blood
of each rat were of the long and slender, or intermediate types.
Subsequently, short forms made their appearance, and after a few
days posterior nuclear forms. In no case was a posterior nucleated
io6
form found in the first day’s count of 200 in any rat. The
posterior nuclear forms made their appearance in the peripheral
blood with considerable regularity, in the following order: —
first A forms, next B forms, and lastly C forms. Thus of the
twenty-four rats, A forms appeared first in thirteen, B forms first
in six. In four, A and B forms appeared together, while in one,
A and C forms appeared together. In none of the rats was a
C form the first type of posterior nuclear to appear.
DAY ON WHICH POSTERIOR NUCLEAR FORMS FIRST
APPEARED
The earliest day of the disease on which posterior nuclear forms
were first seen in the peripheral blood in this count was the seventh,
the latest on which they first appeared was the thirteenth. The
average day of their appearance in the twenty-four rats was the
ninth day of the disease. From the time of their first appearing,
posterior nuclear forms were usually to be found up to the death
of the animal. It was observed that the posterior nuclears
increased not only actually as the disease progressed, but also
relatively to other forms of parasite. The greatest number of
posterior nuclear forms in any 200 counted occurred on the
day of death in fourteen out of the twenty-four rats, on the day
preceding the day of death in seven rats, two days before the day
of death in two rats and previous to this day in the remaining rat.
From the enumeration of the parasites in films from the infected
animals by this preliminary count, the results obtained were as
follows: —
(1) The first forms of parasite found present in the peripheral
blood were long and intermediate free flagellated forms.
(2) Short forms appeared later.
(3) Posterior nucleated forms only appeared after the disease
had developed somewhat.
(4) Posterior nucleated forms increased in numbers from the
time of their appearance, both actually and relatively to other forms
of trypanosome.
(5) Of the posterior nuclear forms those with the nucleus near
the centre of the trypanosome appeared first, those with the nucleus
io7
near the blepharoplast last, while the forms with the nucleus
between those two extremes made their appearance at an intermediate
stage of the disease.
THE CONSTANCY OF APPEARANCE OF POSTERIOR
NUCLEAR FORMS IN THESE EXPERIMENTS
Laveran 8 states that the morphological peculiarity of
T. rhodesiense (posterior nuclears) in rats and mice is not constant.
In these experiments, comprising twenty-four rats, there was no
exception. Posterior nuclear forms were found in all cases in the
preliminary count. Two rats only did not present the advanced
form (C form) during this count, but this form was easily found
on further search.
2. Second count . Two rats out of each group, each having a
good infection, were chosen for the purposes of this count. A
thousand trypanosomes were enumerated whenever possible in each
daily film, and the posterior nuclear forms noted as before. The
results of this more laborious procedure confirmed the conclusions
arrived at as the result of the first examination, the earliest day on
which posterior nuclears were first found being the sixth to the tenth.
It appeared also that C forms occurred in greater proportion in
those animals which lived the longest time, and that within the
group of posterior nuclear forms the C forms generally increased
relatively to the A and B forms towards the end of the disease.
Table II gives an illustration of some of these points; it gives the
number of posterior nuclear forms of the three kinds per thousand
of trypanosomes during the course of the disease in two rats.
POST-MORTEM RELATIVE INCREASE OF POSTERIOR
NUCLEAR FORMS
In several of the rats from which films were taken after death
it was observed that the proportion of posterior nuclear to other
forms increased considerably. This phenomenon appears to be
related to the observation that frequently after death the ‘short
stumpy 9 forms of parasite in T. rhodesiense resist the processes of
disintegration in the blood of the dead host better than the long
forms. Bevan and MacGregor 9 drew attention to this fact. It
has also been recorded by Swellengrebel 10 in the mouse, and by
Blacklock 11 in the rat.
io8
Taw. i II. — To thow the date* of appearance of poaterior nuclear formi of various grade* in
the peripheral hood of two rats infected with T. rbtdtsienu, — per 1000 trypanosome*.
Rat 2230 C
•
: __ ■
Rat 2231 A
Date
A
B
C
A
B
C
Oct. 4. ...
,-
1 _
(Inoculation)
5 ...
—
—
—
—
-
„ 6 ...
—
—
—
—
—
-
.. 7 ...
—
—
—
—
—
-
„ 8 ...
—
—
—
—
-
9 - -
i
—
—
•_
—
-
i? ...
6
3
—
1
—
-
ii ...
-
2
—
*
—
-
„ 12 ...
• _
—
—
4
2
-
- «3 ....
l
— 1
1
—
—
—
m *4
i
2
4
1
—
n 15 *
—
1
1
1
6
1
, 2
1
„ 16 .J
4
3
' 3
10
i 5
4
n 17 -
l
—
1
8
1
1
„ i8 ...
14
12
6
7
1
5
- 19 —
6
2^
21
—
| -
[
.. I9t ...
3*
31
22
-
1 —
i
• Lett than
loco
counted on film.
-
t After death of rat.
THE SIGNIFICANCE OF POSTERIOR NUCLEAR FORMS
IN r. RHODESIENSE
Many explanations have already been advanced to account for
these forms of parasite. Bevan 12 states that the presence of short
forms of trypanosome with the macronucleus slightly posterior to
the centre is a common feature in many species, and especially in
trypanosomes undergoing degeneration or taki ng on the resistant
form in the presence of adverse or unusual conditions. This
io9
explanation does not suffice to explain the typical posterior nuclear
forms in T. rhodesiense , because in this species it is not a case of
the nucleus being slightly posterior to the centre merely. There are
certainly many forms which conform to this description, but the
typical posterior nuclear forms do not fall under this category;
they are something much more definite. As regards the suggestion
that they are the product of degeneration, it is difficult to explain
why they are not found when T. gambiense degenerates. As
previously stated, there is some evidence that such forms are
capable of great resistance to the processes of disintegration in the
cadaver. It appears improbable that the same phenomenon,
4 posterior nuclears/ could be evidence at one and the same time of
degeneration and resistance: it is possible, of course, that the
trypanosomes may assume this arrangement of the nucleus in
response to the demands of an unsuitable environment, and that
subsequently if the environment continues to be unsuitable they
degenerate, still retaining this arrangement of the nucleus. But
the study of T. gambiense under conditions where the parasites are
obviously degenerating has not, up to the present, led to the
discovery of such forms.
ABERRANT FORMS
Bruce (loc. cit.) refers to the posterior nuclear forms seen by
him in the T . rhodesiense of Nyasaland as ‘ aberrant 9 forms. In
view, however, of the fact that they are so constant in their
appearance in this strain and form an integral part of it, it seems
hardly justifiable to treat them merely as ‘ aberrant * forms.
DUE TO TECHNIQUE
It has been stated that they may be due to methods of taking
films, fixing or staining, but so many observers have noted their
appearance, using different methods of treatment for the films, that
it seems reasonable to discard this view.
In seeking for an explanation of their presence, it is essential
to discover, if possible, whether or not they ever occur in
T. gambiense. Up to the present, so far as one is aware, they
have not been described in this trypanosome. It may be argued
iio
that the number of T. gambiense strains which has been examined
is small. If, however, it can be established that they are entirely
absent from T. gambiense , it will be difficult to attribute their
presence in T . rhodesiense to such changes as degeneration, or to
resistance, or to relegate them to the class of ' aberrant * forms.
If further examination of strains of T . gambiense fails to reveal
their presence, it must be concluded that these forms constitute an
important distinction between the two parasites T. gambiense and
T. rhodesiense. The fact that such forms are found in animal
strains does not diminish their claim to attention, since animals
are known to be infected with human trypanosomes. The argument,
that if so many animal strains were capable of infecting human
beings there would be a great amount of human trypanosomiasis
in regions where it does not at present abound, is not conclusive.
It has to be definitely proved that there is actually no human
trypanosomiasis in such animal-infected regions. The experience
in Nyasaland, and in Rhodesia, Northern and Southern, renders it
necessary to be cautious in considering a region free from sleeping
sickness. Even if human cases were rare, it may fairly be argued
that we do not by any means fully understand the factors that
govern the successful infection of human beings.
FLY VARIATION
If we regard T . rhodesiense simply as a variation of the human
parasite of the Gambia, it is possible that the explanation of their
occurrence is to be sought, not so much in the conditions of their
environment in the blood of human beings and in that of inoculated
animals, as in other factors, for example, the transmitting agent,
the fly. Against this hypothesis we have the facts that although
T. vivax and T . pecorum are each known to be transmitted by both
G. palpalis 13 and G. morsitans , 14 they do not appear to be modified
morphologically by the difference of carrier.
It would be of considerable interest to ascertain whether in cases
where T . gambiense is transmitted by G . morsitans such posterior
nuclear forms are to be found in the blood of infected animals.
A
Ill
MULTIPLE INFECTION
The possibility of there being an infection in human beings
with more than one species of trypanosome suggests itself. In
favour of this one might adduce the facts that not only do animals
in nature frequently suffer from mixed infections, but also that
certain species of fly have proved themselves capable of
transmitting more than one species of trypanosome. It seems
probable that human beings are susceptible to mixed infection.
The proof and demonstration of a double infection, given that the
human being harboured species which were each universally
pathogenic to laboratory animals would, with the means at present
at our disposal, be practically impossible. In this connection one
might refer to the original Gambian Horse Trypanosome described
by Dutton and Todd 15 ; in this case infection was considered to be
caused by a single species of trypanosome, a view which is not now
generally accepted. It is only where the pathogenicity of the
trypanosomes taking part in a mixed infection is widely divergent
as regards laboratory animals that proof of the presence of more
than one sepcies of parasite can be given.
Against the hypothesis of mixed infection in the case of
T . rhodesiense is the evidence derived from transmission experi¬
ments. Kinghorn and Yorke, 16 transmitting T. rhodesiense by
means of Glossina morsitans bred in the laboratory, found posterior
nuclear forms in all animals which became infected. This implies,
accepting for a moment the mixed infection hypothesis, that the
flies which became infected acquired and passed on the mixed
infection in each case. This is, perhaps, the strongest argument
against the hypothesis: A further argument against the possibility
is somewhat of the same order, namely, the results of infection by
culture of T. rhodesiense , which Bayon 17 records.
CONCLUSIONS
1. Posterior nuclear forms first appear in the blood of rats
infected with T. rhodesiense from the sixth to the tenth day of the
disease, taking a count of a thousand trypanosomes.
2. They increase in numbers in the later stage of the disease.
3. They increase relatively to other forms of trypanosome.
112
4. They cannot be explained as the result of either faulty
technique or degeneration.
5. They show definite powers of resistance to disintegration in
the cadaver of the animal host.
6. The presence of posterior nuclear forms may be due to: —
(a) The occurrence of such forms as a constant constituent of
certain strains.
(< b ) A mixed infection,
( c ) Certain unexplained influences in the blood environment,
affecting the parasites.
( d ) The transmitting agent.
7. There are, at present, stronger arguments against the last
three explanations of their presence than against the first.
RBPBRENCBS
1. Proc. Roy. Soc., 1910, Series B, LXXXIII, No. B 561, pp. 28-33.
2. Proc. Roy. Soc., 1911, Ser. B, LXXXIV, pp. 136-160.
3. Diary P.M.O. Nyasaland Protectorate, Part xvii, * Sleeping Sickness,* p. 7.
4. Proc. Roy. Soc., 1912, Ser. B, LXXXV, No. B 581, pp. 428-33.
5. Journal Trop. Med. and Hyg., 1912, July 1, XV, No. 13, p. 193.
6. Brit. Med. Journal, 1912, Aug. 31, p. 473.
7. Brit. Med. Journal, 1912, Oct. 19, p. 1057.
8. Vide Sleeping Sickness Bull., 1912, Vol. IV, No. 33, p. 3.
9. Jour. Compar. Path, and Ther., 1910, June, pp. 160-167.
10. Centralblatt fiir Bakteriologie, 1911, Heft 3, pp. 103-206.
11. Ann. Trop. Med. and Parasit., 1912, May, VI, No. 1 B, pp. 55-68.
12. Letter, Sleep. Sick. Bull., 1912, Vol. IV, No. 38, p. 214.
13. Report Sleep. Sick. Com. Roy. Soc., No. XI, pp. 136 and 175.
14. Communication, Dr. Warrington Yorke.
15. Memoir XI, Liver. School Trop. Med.
16. Ann. Trop. Med. and Parasit., 1912, VoL VI, No. 1 A, p. 3.
17. Vide Sleep. Sick. Bull., 1912, Vol. IV, No. 40, p. 319.
IIJ
ANIMAL TRYPANOSOMIASIS IN THE
LADO (WESTERN MONGALLA) AND
NOTES ON TSETSE FLY TRAPS AND
ON AN ALLEGED IMMUNE BREED OF
CATTLE IN SOUTHERN KORDOFAN
BY
ANDREW BALFOUR, C.M.G., M.D., B.Sc., F.R.C.P.E.,
D.P.H.,
DIRECTOR, WELLCOME TROPICAL RESEARCH LABORATORIES, GORDON COLLEGE, KHARTOUM
(j Received, for publication 24 February , 1913)
Plates IX, X
When, some years ago, the Sudan Government took over the
region known as the Lado Enclave, which now stretches from the
sixth parallel of North Latitude southward to Lake Albert and
extends from the Bahr-el-Jebel on the east some 120 miles inland to
the frontier of the Belgian Congo, it found itself in possession of an
area infected with sleeping sickness. Since that time a great deal has
been done by Captain Thompson and Lieut. Ranken, R.A.M.C.,
in studying this disease and to combat it both prophylactically and
therapeutically, but, so far, little attention has been paid to the
forms of animal trypanosomiasis present in this part of Africa.
The following short and, of necessity, incomplete paper is intended
as a preliminary contribution to the subject, and also contains notes
on certain other matters intimately associated either with human or
animal trypanosomiasis.
I. In May of last year I visited the sleeping sickness camp at
Yei, in what is now known as Western Mongalla, the name Lado
having been abandoned, as the old Belgian station of Lado on the
Nile has disappeared. On the way thither, and when at Yei, I had
a few opportunities of examining the blood of transport and other
domestic animals. At Loka, a post intermediate between Rejaf
and Yei, I found mules infected with trypanosomiasis, and at Yei
found a donkey which had been infected when in the fly region of
the Moru district further to the north and west.
Lately Captain R. J. C. Thompson, R.A.M.C., who has done
such good work in charge of the sleeping sickness camp and in
connection with clearance operations, sent me the blood of three of
the transport bulls working on the old Belgian automobile road
which runs from Rejaf on the Nile, through Loka and Yei, to Aba
on the Congo frontier.
He had found these animals infected, and was anxious to know
the type of trypanosome present. We will, therefore, first consider
the infection in the case of these bulls. A mere glance at the films
showed that we were dealing with a trypanosome of the pecorum-
nanum-congolense group. The films had been fixed in absolute
alcohol, and, though rather old, stained sufficiently well with
Giemsa to permit of their being accurately drawn at a magnification
of 2,000 diameters by the aid of Greil’s projection apparatus. This,
combined with Bruce’s method of measuring, is probably the most
accurate means of determining the dimensions of stained trypano¬
somes. The best results are, however, obtained by fixation with
osmic acid. From the three slides 140 parasites were drawn and
measured. Attached is the curve (Chart I) obtained on plotting the
measurements according to Bruce. It will be found to answer very
closely to that for the Sudan T. nanum, shown in Vol. A of the
Fourth Report of these Laboratories (1911, p. 55). The trypano¬
some is small, short, and shows no free flagellum. The following
statement gives the figures in respect to length : —
Minimum Maximum Average
Length in microns ... 9’8 ... 17 ... 126
These dimensions are quite near enough those given by Bruce,
Laveran and Mesnil and other observers for T. nanum to permit of
its being considered as this trypanosome, so far as length goes.
The breadth at the broadest part varied from 1/1 to 2/1, which is
also in keeping with that of T. nanum. The accompanying
microphotographs show sufficiently well the general aspect of the
parasite. PI. IX, fig. 1 shows a typical short T. nanum -like form;
fig. 2 a longer variety with a better marked undulating membrane.
Neither shows any free flagellum. The infection was, in none of
the three cases, very heavy, but I am informed it is proving fatal to
the transport bulls. Captain Thompson also informed me that he
thinks it may be associated with trypanosomiasis in a herd of cattle
n 5
at Mongalla on the east bank of the Nile. I wired to the Governor as
to the locality from which this herd came, but failed to elicit
any definite information. Archibald, 1 however, has described
T. fecorum as occurring in a cow from Bor, which is in the more
Values or
Chart^I. Curve of measurements of trypanosome from transport bull.
northerly part of the Mongalla Province and on the east bank of
the river. Until animal inoculations were carried out, it was of
course impossible to decide accurately the species of trypanosome
with which we were dealing. Accordingly, I wired to Captain
Thompson, asking him if at all possible to inoculate laboratory and
other animals, and to let me know the result. I have now heard
from him to the effect that an inoculated dog showed trypanosomes
in its blood fourteen days after the injection.* It would, therefore,
• * These trypanosomes then disappeared and were not again found, while the dog has
remained throughout in good condition. Fifteen rats which were inoculated died within 48
hours of inoculation from some unknown cause. It would seem, therefore, that the bull
trypanosome may have been T. nanum after all.
seem that we are dealing with T. pecoruniy not T. nanurn.
Certainly fig. 2 shows a trypanosome more of the pecorum than the
nanum type, and also (if we accept the views of Laveran and
Mesnil 2 as to T. congolense being a distinct species) than the
congolense type. As a matter of fact, so far as the measurements
go, they are nearer those given for T. pecorum than for T. nanum.
In any case the positive result of animal inoculation puts the matter
beyond doubt so far as these two species are concerned. (But see
footnote on p. 115.)
If the infection has been derived along the automobile road in
Western Mongalla (late Lado Enclave) the vector is in all
probability Glossina palpalis , as G. morsitans is not found in this
locality, and the only other biting flies in evidence are
Haematopota. It is, however, very probable that these bulls were
infected before going to Western Mongalla, and that some species
of TabanuSy Chrysops y or Stomoxys is the active agent in
transmitting this form of animal trypanosomiasis in the Southern
Sudan. I have mentioned the donkey from the Moru country.
I kept only one film of this animal’s blood, and, as the infection
is very small, have not troubled to draw and measure the parasites.
The trypanosome, however, is undoubtedly of the same type as that
found in the bulls (fig. 3). Again, if we accept Laveran’s view,
I think it will be admitted that it is very like T. congolense.
Considering the history of this trypanosome, it is exceedingly
likely that it is T. congolense , but there is, of course, no definite
proof so far.
As regards the infected mules at Loka, I took films and fixed
them in absolute alcohol, having no osmic acid with me. Later
these films were stained with Giemsa. The infection was somewhat
sparse, and at a magnification of 2,000 diameters it took a long
time to draw even as many as a hundred from different slides.
This, however, was done, and they were measured by Bruce’s
method, the curve being given in Chart II.
The measurements in respect to length are as follows: —
Minimum Maximum Average
Length in microns ... 10 5 ... 28 ... 176
Figs. 4 and 5 show the general aspect of the parasites. It will
be seen that this trypanosome is of the brucei or gatnbiense type
ii7
in that there is a long and comparatively slender form with a free
flagellum, and a short, stout form with hardly any free
flagellum. (PI. IX, fig. 5, shows a form with the blepharoplast just
beginning to divide.) The breadth of the former varies from
i'2 m to 2 ft, of the latter from i jp to as much as $/*, there being
a few very stout and swollen-looking forms. These did not seem
in any way degenerate, but the appearance they presented was
possibly artificial and due to slow fixation permitting of contraction
and broadening. The usual breadth of these forms was from
2‘5fl to 3 fi.
A marked feature of this trypanosome was the frequent
occurrence of an unstained area, usually of a spherical shape, in the
neighbourhood of the blepharoplast. This is, of course, sometimes
Coast II. Curve of measurements of mule trypanosome.
seen in T. gambiense and other species, but I have never known it
so frequent or well defined as in this trypanosome. It was
specially evident in the short, stout forms, but was occasionally
seen in the forms showing a long free flagellum (figs. 4 and 5).
Possibly it is merely the product of a faulty fixation. It will be
noticed from the curve that a few dwarf or stumpy forms were
present. Whether these indicate that there is a double infection,
or whether they are like the so-called ‘ tadpole * forms which have
been described in T. dimorphon infections, I am unable to say.
One of the sick mules accompanied me on my return journey from
Loka to Rejaf. At the latter place I had a couple of gerbils on
board our floating laboratory. These animals (fi. pygargus) are as
a rule very susceptible to trypanosome inoculation,* but though
* Infection invariably remits after inoculation with 7 . gambit nst, 7 . tvsnsi, 7 . brucei
(ptceudt), and 7 . pecarum.
I gave each of them a large dose of citrated blood from the sick
mule neither developed any infection.
Under these circumstances, I am unable to express any opinion
as to the species which this trypanosome represents. The curve,
Chart II, is certainly rather peculiar, there being two apices. It may
be that a sufficient number of trypanosomes has not been measured,
or, again, it is possible that had the fixation been better the staining
of the free flagella might have been better also, and the two apices
might then have been compounded into a single apex at or about
igfi. Even in this case, however, it would not have answered very
closely to that given by Bruce for the Uganda strain of T. brucei.
I suggest that it may be the same pleomorphic trypanosome
{Trypanosoma, sp. ?) found by Kinghom and Yorke 3 in a
bushbuck. The mule transport had been working in the
southern Bahr-el-Ghazal before coming to Western Mongalla,
so that it is possible the infection was derived in the former
province, where but little is yet known regarding the different
strains of trypanosome affecting domestic animals. In the past
there has been free communication between what was the Lado
Enclave and both Uganda and the Belgian Congo, so that, in all
probability, various trypanosome infections of bovines and equines
have been introduced into Western Mongalla. It is very desirable,
as has been shown by the work of Kinghom and Yorke in many
reports, notably those cited, 3 4 as also by Duke 6 and by numerous
French observers in West Africa, to determine accurately the
different species of trypanosome affecting domestic stock and their
insect vectors. These short and incomplete notes may be of some
help to those whose duty it will be to work out the problems on the
spot, for there is now a good laboratory at Yei, and Lieutenant
Ranken intends to extend his researches, both to the disease in
domestic animals and to the question of the presence of trypano¬
somes in big game. The region is so remote and difficult of access
that it is well-nigh impossible to carry out the work satisfactorily in
Khartoum.
II. In the British Medical Journal for July 6th, 1912, I 6 drew
attention to the subject of tsetse fly traps. I have now had large
traps constructed by the Andr6s Maire Company. These (PI. X,
fig. 6) are not exactly what I intended. Indeed, they differ very
ii9
little from the cotton moth trap invented by this firm, but both
Mr. King and I think they may serve the purpose of trapping
tsetses, provided it is found the flies will enter them.
As regards methods of attraction, the following measures will
be tried: —
1. Soaking the central bands or wicks in a mixture of water
and either human or animal sweat.
2. Placing a live animal in the trap—a suggestion made by
Captain O’Farrell.
3. Soaking the central bands in citrated blood, and at the
same time hanging up in the trap a piece of fresh meat
from ox or buck with skin still adherent.
4. Soaking the bands as above and having in addition a tube
of citrated blood arranged according to the ingenious
device of Rodhain and his colleagues. 7 If desired,
the blood in this tube may be poisoned.
It is hoped that one or other of these measures may be effective.
Lieutenant Ranken will shortly take two of these traps with him to
Yei, and carry out the necessary experiments.
III. Fig. 7 is a photograph of a full-grown bull of the small
black breed found in Southern Kordofan. It was kindly sent me,
at my request, by Major R. S. Wilson, Governor of the Nuba
Mountains Province.
This breed is said to be immune to trypanosomiasis, and is the
only breed which can live in the infected Koalib area, where
G. morsitans abounds and conveys an infection due to what
I believe is T. brucei. It is interesting to note that an apparently
similar breed exists in Northern Nigeria. Pollard 8 reports that
There is in the Munshi Division, and in the northern part of the
province, a small black breed of cattle which is apparently immune
to tsetse. At any rate, these cattle can be kept in the Munshi
district, where no horses can live and where imported Fuhlani cattle
all die. I have not yet obtained any blood-films from these
animals.*
The Kordofan breed may, of course, only be immune to one
species of trypanosome, and, where only one specimen of these cattle
is available, it does not seem desirable to make experiments in this
120
direction. On the other hand, it will be interesting to note the
effect of the blood serum of this bull on various trypanosome
strains. Some work along this line has been commenced, and will
be continued as circumstances permit. At present it is too early to
make any statement regarding it.
I am indebted to Mr. G. Hunt, of the Engineering School of the
Gordon College, for kindly plotting the curves for me. The
photographs are the work of Mr. George Buchanan.
REFERENCES
1. Archibald, R. G. (1912). A Trypanosome of Cattle in the Southern Sudan. Journal
of Comparative Pathology and Therapeutics, XXV, 4, pp. 292-297.
2. Laviran, A. Sc Mksnil, F. (1912). Trypanosomes et Trypanosomiases. 2nd edition.
Masson, Paris.
3. Kinohorn, A. Sc Yobkx, W. (1912). Trypanosomes infecting Game and Domestic Stock
in the Luangwa Valley, North Eastern Rhodesia. Annals of Tropical Medicine and
Parasitology, VI, pp. 301-323.
4. Kino horn A. Sc Yoan, W. (1912). Further Observations on the Trypanosomes of
Game and Domestic Stock in North Eastern Rhodesia. Annals of Tropical Medicine
and Parasitology, VI, pp. 483-493.
5. Dun, H. L. (1912). The transmission of Trypanosoma nanum , Lave ran, and Some
observations on Trypanosoma ptcontm , Bruce, and 7 . uniform*, Bruce. Proceedings of
the Royal Society, LXXXV, pp. 4-9 and 554-561.
6. Balfour, A. (1912). Notes on Sleeping Sickness. British Medical Journal, U, pp. 10-11.
7. Rodhain, J., Pons, C., Vandrnbranden, J., Sc Brquarrt, J. (1912). Contribution au
M^canisme de la Transmission det Trypanosomes par les Glossines. Arch. f. Schiffs.
u. Trop. Hyg., Nov., XVI, ai, pp. 732-739.
8. Pollard, J. (1912). Notes on the Tsetse Flies of Muri Province, Northern Nigeria-
Bulletin of Entomological Research, III, 2, pp. 219-221.
122
EXPLANATION OF PLATES
Trypanosomes magnified 2,000 diameters approximately.
Plate IX
Fig. 1. Trypanosoma pecorum ? From bull. Note short, stout
forms.
Fig. 2. T. pecorum ? Long form. From bull.
Fig. 3. T. congolense ? From donkey.
Fig. 4. Trypanosoma , sp. ? From mule.
Fig. 5. Trypanosoma , sp. ? From mule.
Annals Trop. MtJ. S3 Parasitol., Vo\. VI1
PLATE IX
c. Tinting A- Co., Ltd., Imp
Fig. 5
Plate X
Tsetse fly trap.
Pigmy bull of black breed from Southern Kordofan, said
to be immune to ‘ fly.’
Annals Trap. Med. & Parasitol., Vo!. VII
PLATE X
inling < 5 - Co., Ltd., Imp.
125
SANITATION ON THE PANAMA CANAL
ZONE, TRINIDAD AND BRITISH GUIANA
BY
DAVID THOMSON, M.B., Ch.B. Edin., D.P.H. Cantab.
(i Being part of the Report on the Malaria Expedition to
Panama , 1912)
(Received for publication I March, 1913)
PLATES XI, XII, XIII
CONTENTS
PAGI
Introduction . 125
COMMENCEMENT OF SANITATION ON THE ISTHMUS . 128
Sanitation in Colon . 128
Sanitation in Panama Town. 129
Sanitation on the Zone or the Canal—Anti-Malaria Work ... 130
Anti-Yellow Fever Work . 136
Sanitary Measures adopted to safeguard against Plague . 137
Measures against Aneti.ostomiasis . 137
Anti-Typhoid and Anti-Dysentery Work and General Sanitation ... 138
The Hospitals on the Canal Zone 138
Results of Sanitation on the Canal Zone . 140
Sanitation in Trinidad . 141
Sanitation in British Guiana . 143
References 146
Explanation of Plates 148
INTRODUCTION
In September, 1912, I was sent on an expedition to the Panama
Canal Zone by the Liverpool School of Tropical Medicine to study
certain malarial problems with Dr. James, Chief Assistant
Physician to the Ancon Hospital. The American Canal Commission
kindly allowed me to stay in the Ancon Hospital, and it was there
that we conducted our researches. Although occupied for the most
part with special research, I was nevertheless able, during my three
months* stay, to see most of the sanitary work going on. I received
much information on the subject from the medical men in the
126
hospital, and Colonel Gorgas, Chief Sanitary Officer, Colonel
Phillips and Mr. Le Prince, Chief Sanitary Inspector, were always
willing to give me all the information that I required regarding their
organisation and the methods employed. Moreover, I walked over
most of the line of the Canal Zone with Dr. Orenstein, Chief
Assistant Sanitary Inspector, who was most kind in showing me
everything that he possibly could regarding the anti-malaria work.
After all that I have seen there, I feel compelled to express much
admiration regarding the great system of sanitation, and nothing
was apparently left undone in the eradication of malaria and other
diseases in general. There was no apparent defect in Colonel
Gorgas’s sanitary administration. All trusted him in his work
with implicit confidence, and the highest authorities gave him power
to do what he considered best in great as well as in small measures.
The money required was granted with the full belief that it was
necessary for the good health of the American inhabitants, and for
all workers on the Canal. They realised that good health was to be
obtained at all costs, not only for its own sake, but in order to
enable them to finish the great undertaking which they had
commenced. The Americans, in the Canal Zone at least, are
convinced that sanitation 1 pays.* They had before them the great
lesson of the disastrous failure of the French Canal diggers, and
they have learnt that lesson well. No one who had not lived on
the Canal Zone before the Americans took it over could possibly
realise fully the great changes that have been brought about. The
Ancon cemetery, which is a veritable forest of tombstones erected
chiefly to French victims (PI. XII, fig. 9), helps one to realise
that in bygone times the risk of death was very high. In an
unhealthy tropical country like West Africa, there is always a
feeling of insecurity. People are constantly sick, many die, and
one feels that perhaps one’s own turn is coming next. Mosquitos
abound, one is constantly bitten; mosquito nets may be used,
quinine may be taken every day, and even though one is careful to
drink only filtered or boiled water, yet a feeling of security against
malaria and dysentery is never felt. So it was in Panama less
than ten years ago. Formerly mosquitos were a plague on board
vessels lying in Colon harbour, but since the American occupation
that is all changed, and now no mosquitos come aboard. I
127
arrived on the Canal Zone fully equipped, among other things,
with a mosquito net and plenty of quinine which are always so
necessary in tropical expeditions, but soon realised that both the
mosquito net and the quinine were not needed. There were no
mosquitos about, and every American house was mosquito-proof.
My three months’ stay covered the latter part of the rainy season
and the commencement of the dry season, yet during all that time
I feel sure that I was not bitten by a single mosquito,* and in
consequence I did not take one single grain of quinine. The water
supply was good, and was frequently examined bacteriologically.
Healthy European women and children were met everywhere, no one
seemed to be ill, and one felt as secure from disease as anywhere
in temperate climates. I have been in Sierra Leone, Forcados,
Bonny and Old Calabar on the West Coast of Africa, also in
Trinidad, British Guiana and Jamaica. In all of these places
I was bitten by mosquitos, even though my stay had been only for
a few days. In all I had to take quinine as a prophylactic against
malaria. In these places, moreover, there are not so many white
women and children seen as on the Canal Zone at Panama. In
West Africa there are no white children and few white women, for
the simple reason that it is too unhealthy. I think I have said
enough to indicate, without going into the statistics of death-rate,
etc., that Panama, once notorious as a white man’s grave, is now as
healthy as most temperate countries, and the favourable reports
which may be read concerning this matter are in no way exaggerated.
I have known several traders whose occupation has brought them
many times to the Canal Zone during the last ten to fifteen years,
and all alike marvel at the great changes which have occurred for the
better in so short a time. Most of the authentic reports on the
improved health in Panama have been gleaned from the statements
made by Colonel Gorgas. He has in no way exaggerated the
results of his sanitary work. Again, it must be remembered that
this greatest achievement in tropical sanitation, which the world has
yet seen, reflects great credit, not only on Colonel Gorgas and his
staff, but also on the highest officials on the Canal Zone and on
the American Government who have supplied the money without
* Mofquitot are fairly common still at Frijoles and Monte Lirio.
128
complaint and without putting obstructions in the way, actively or
passively, and who have been among the first to realise that
thorough sanitation in the tropics ‘ pays financially.’
COMMENCEMENT OF SANITATION ON THE ISTHMUS
Plans for the sanitation of Colon and Panama formed part of
the discussion at the American Commission of 1899-1901. These
towns, although situated on the Canal Zone, belong to the Republic
of Panama. In January, 1904, the quarantine of Colon and
Panama was turned over to the United States, and in June of that
year the permanent sanitary organisation was established with
Colonel W. C. Gorgas as head and Dr. H. R. Carter, a yellow
fever expert, as the Director of Hospitals. The work was hampered
by scarcity of supplies, notably copper wire screening, which could
not be purchased in the United States in large quantities. An
epidemic of yellow fever, lasting from July, 1904, to December.
1905, accelerated the delivery of supplies, and made it necessary
to expedite the sanitation work, lest the force slowly organising be
depleted. There were 246 cases and 84 deaths, all of which were
among the non-immunes who had come to the Isthmus on account
of the Canal work. The Americans commenced sanitation,
therefore, before attempting any operations on the Canal. They
realised that it would be advantageous to make the place healthy
before bringing the white population there. They were pioneers in
that they built every house mosquito-proof.
SANITATION IN COLON
This town is situated at the Atlantic end of the Canal. It has
a population of 18,000. In 1904 it had a population of 10,000,
and about 9,000 of these lived in shanties built on piles. At high
tide the houses were surrounded with water, so that no one could
walk along the streets without danger of falling into the mire
(PI. XI, figs. 1, 3). Behind the town was a large mangrove
swamp. There was no proper water supply, and no sewage system.
Mosquitos abounded, and malaria and other diseases ran riot. Since
then the town site has been filled in by the Americans. It has been
raised several feet, so that it is now dry, well paved (fig. 2),
129
clean, well drained and healthy, with a modern sewage system and
water supply. The Panama Republic and the Panama Railway
Company are paying for the work.
SANITATION IN PANAMA TOWN
Panama is situated near the Pacific end of the Canal, and has
a population of about 37,000. As already stated, there was a
yellow fever epidemic raging there from July, 1904, to December,
1905. This, however, was the last of the disease in that town. The
Americans in a little over a year eradicated it entirely, and the
conditions are now such that it could hardly exist there again.
This alone is a triumph which must have already paid financially
many fold. Panama Town had previously been an endemic yellow
fever centre for centuries, and in a little over one year the disease
was stamped out and not a single case has occurred there since.
The methods adopted to control the epidemic were isolation of all
cases as well as suspected cases, in mosquito-proof quarters, and
thorough fumigation of all houses which had cases or suspected
cases, in order to kill the mosquitos in them. At the same time a
campaign was started to destroy the breeding places of the
Stegomyia fasciata . At that time every house in Panama had roof
gutters and water tanks or barrels, this being almost the sole water-
supply of the town. There was no proper drainage or sewage
system. The streets were unpaved, without proper gutters, and in
the rainy season became a hopeless quagmire (fig. 4). The
conditions, therefore, favoured greatly the breeding of the
Stegomyia fasciata. The Americans employed no makeshift
methods in remedying these defects. They brought in a proper
pipe-borne water-supply from a reservoir. They installed into
every house modern closets, with a pipe-borne sewage system. They
paved and guttered every street (fig. 5), and removed as far as
possible all the roof gutters from the houses along with the water
barrels. They enforced strict sanitary regulations on the
inhabitants of the town, and established a health department to see
that these regulations were carried out. In addition, the ground
floors of the houses were rendered uninhabitable to rats by
cementing f and cement gutters were constructed alongside the pave-
ments in order to catch the rain from the gutterless eaves of the
houses. These were the drastic measures enforced on the
inhabitants of Colon and Panama. The Americans lent the money
and supplied the labour. It cost the two towns about £455,000.
They are paying it back with interest through water rates which are
collected directly by the United States. The inhabitants who had
lived there in a slovenly way without interference, as their
forebears had done for centuries, were naturally very angry at this
apparent tyranny, but already they are becoming satisfied. Those
who have families have already noticed an unprecedented freedom
from disease among their children. Plague, typhoid fever,
dysentery and malaria are no longer to be feared, and now they
realise that what was done was good. The sanitation of Colon and
Panama is to-day as up-to-date as in any town in Europe. The
water-supply is examined very frequently by expert bacteriologists.
The quarantine at Panama and Colon is most rigid. Guayaquil in
Ecuador is an endemic yellow fever centre, and is only about four
days’ sail from Panama, hence a very strict quarantine is necessary.
In fact, if the Ecuadorians do not soon take steps to make their
town sanitary, it is highly probable that the Americans will force
them to do it, as it is a constant menace to the health of Panama.
Trinidad and all Venezuelan and Colombian ports are under
quarantine for yellow fever, though I believe that since my return
Trinidad has been removed from the list.
SANITATION ON THE ZONE OF THE CANAL—ANTI-MALARIA
WORK
On the Canal Zone the most prevalent disease has been malaria,
and much of it is pernicious malaria. The sanitary measures
adopted were, therefore, principally anti-mosquito measures.
Countless swamps and Anopheles breeding places existed along the
line of the Canal. A huge swamp lay behind Colon, and extensive
swamps also surrounded the town of Panama. After enormous
labour, these have been filled up by material excavated from the
Canal. Two methods have been adopted in filling the swamps.
One consists in dumping material (rock and earth) into them from
railway waggons. The other method, known as a ‘ hydraulic fill ’
(PI. XII, fig. 8), is to force into them a liquid stream of mud
and sand through pipes leading from large suction dredgers
employed in deepening the Canal. In many cases these pipes are
laid long distances, even a mile or more distant from the Canal.
I have seen swamps filled in this way to the depth of many feet,
so that only the tops of the trees which grew in these swamps are
now visible (fig. 8). Large unhealthy areas have been obliterated
in this way. The present town of Balboa on the Pacific entrance
to the Canal is being built on one of these filled-in swamps.
Strictly speaking, however, these huge undertakings can hardly be
considered as anti-malaria work. In describing this, I cannot do
better than give an abstract of a paper by Colonel Gorgas (1909)
on the subject.
He states that the anti-malaria work on the Isthmian Canal
cannot be well understood unless preceded by a brief description of
the sanitary organisation as it bears upon this work. The bulk of
the activities of the Department of Sanitation on the Isthmus has
no immediate bearing upon sanitation, namely, that which deals
with religious instruction, care of the sick, care of the insane,
of lepers, street cleaning, garbage collection, etc. The Department
of Sanitation spent in 1908 some two million dollars, but only
five hundred thousand dollars (£100,000) of this was spent on pure
sanitation. The American Canal Zone consists of a strip of land
ten miles wide, of which the Canal is the centre. This strip extends
about forty-five miles in length from North to South. The
population to be protected against malaria consists of about 50,000
labourers and their families, and is scattered all over this
five hundred square miles, though they are principally collected
along the line of the Canal, more particularly into some forty camps
and villages near this line. The temperature, rainfall, and
character of the terrain are all excellently suited for the breeding
of Anophelines all over this territory (fig. 7). The rainfall
averages over 100 inches yearly, and though there are four months
in which there is practically no rainfall, there is enough water for
the Anophelines to breed freely during these four months. During
the five years of occupancy of the Isthmus up till 1909, 250,000
people have resided in this zone, and these have been located
principally in places formerly unoccupied along the line of the
Canal, and as the villages are intended for only temporary
I 3 2
occupancy, the conditions are a good deal like those of an army
going into a new country, and increase the difficulty of sanitation
enormously.
The anti-malarial measures consist: —
1. In destroying the habitat of the Anophelines, during their
larval stage, within a hundred yards of dwellings.
2. Destroying within the same area all protection for the adult
mosquito.
3. Screening all habitations so that mosquitos cannot have
access.
4. Where breeding places cannot be destroyed by draining, use
is made of crude oil and larvicide.
These measures are given in the order in which they are
considered important.
For the purpose of carrying into effect these measures, the
five hundred square miles of territory have been divided into
seventeen districts. These districts are under the charge of a
Chief Sanitary Inspector, who has in his office the necessary clerical
force and three assistants. One of these assistants is especially
competent in the life-history and habits of the mosquito, another
in knowledge of ditching, tile-draining, etc., and the third in
knowledge of general executive work. Each of the seventeen
districts has a District Sanitary Inspector in charge. Each District
Inspector has a sufficient force of labourers (forty to fifty) to do the
necessary ditching and draining. A force of carpenters is required
to keep the screening in repair, and there are one or two quinine
dispensers who are kept constantly going around giving doses of
quinine to those who desire it. It is not attempted to enforce
prophylactic quinine. The Chief Sanitary Assistants are kept
constantly going over the work advising and instructing the District
Inspectors. The District Physician sends in daily to the central
office a report of the number of cases of malaria and the number of
the employees from which these cases come. This report is made
up weekly in the central office. A copy is sent to each District
Inspector, and he is held responsible for any excess of malaria in
his district. If the admission rate for malaria during the week
rises above i| per cent, something is considered wrong, and the
assistants of the Chief Sanitary Inspector are sent to look over the
133
ground to try to discover the cause. The District Inspector, for the
purpose of doing away with the breeding places of larvae, puts
down tile drains wherever they would be suitable. This is
considered the most effective and economical form of drainage.
After it is laid down it requires no more attention. There is no
breeding place left for mosquitos, as no water whatever is exposed
at the surface. A horse mower or scythe can be used for cutting the
grass over it. Where tile-drainage cannot be used, open concrete
ditches are put down. The first cost of this is nearly as great as
that of tiling, and a certain amount of labour is necessary to keep
the ditches clear. They must be swept out once a week. If the
ground cannot be drained in either of the above ways, open ditches
are used. This is the least effective and most expensive form of
drainage. In Panama they fill up rapidly with grass, and have
to be cleaned out about once in two weeks. They are always
breeding places for mosquitos, and have to be constantly treated
with oil or larvicide.
For the purpose of doing away with places which will harbour
the adult mosquito, the inspector clears the ground of brush and
grass for a hundred yards around the place to be protected. Where
the locality is to be occupied for a year or more, it is more
economical to grade the ground and plant grass, which can then be
kept down with a horse mower or scythe. A limited amount of
shrubbery and a few trees about a dwelling are not objectionable.
The inspector keeps the wire screening in repair by constantly going
over it with a force of carpenters. Good wire should last on the
Isthmus at least three years. The inspector uses crude petroleum or
larvicide in such places as cannot be drained, and in temporary
pools caused by construction or at temporary camps where it would
not be economical to drain. Usually when a new location is
occupied the malaria rate is high, frequently as high as 25 per cent,
a week, but always in the course of a month or two, when the
ground is drained and the brush cut, this drops to a
rate somewhere about 1 per cent. The above methods could be
applied to a considerable extent to military organisations. Where
troops remained at one camp for a week or longer, it would be
practicable to clear and drain the ground. It is most important that
the Sanitary Inspector should attend to the work himself. The men
! 34
who do the ditching and brush-cutting, etc., should be immediately
under his control, and he should be held responsible for the proper
performance of the work, or in other words, the work must be
constantly supervised by a man who has a considerable knowledge
of these anti-malarial measures. Prophylactic quinine is looked
upon as an important measure, and is offered in three grain doses
to all employees who will take it. It is placed on the table in all
the messes in liquid and pill form, and one to three dispensers are
employed in each district who go around the various villages.
Fuller details may be had from articles by the following
authors:—W. C. Gorgas(i904, 1906, 1907, 1908, 1909, 1910, 19 12 )*
A. J. Orenstein (1911, 1912), J. A. Le Prince (1908), and
S. T. Darling (1910, 1912); also from the Reports of the
Department of Sanitation of the Isthmian Canal Commission by
W. C. Gorgas, and from the Canal Record.
The anti-malarial work, therefore, on the Canal Zone consists
simply in carrying out thoroughly and in a straightforward manner
the anti-malarial principles laid down by Ross soon after his
discovery of the transmission of malaria by the Anopheles mosquito.
Yet simple though these principles seem, the methods and
organisation of Colonel Gorgas deserve the most careful study. His
system has produced most remarkable results in a country which
was exceedingly unhealthy and in which the difficulties were great
and many, and although the construction of the Canal brought
unlimited labour and material to his aid, yet it also greatly increased
his difficulties, because it is well known that malaria tends to
increase greatly where land excavation is going on, on account of
the innumerable new breeding pools which are produced by these
excavations. This alone necessitates the constant inspection and
tackling of ever-changing conditions. The working population is
constantly changing its location, and this also involves many
difficult sanitary problems and increase of sanitary work. The
working population on the Canal moves about so much that these
methods are even suitable, as Colonel Gorgas has said, for military
organisations in the tropics. It seems to me that the keynote of the
success is thoroughness in every detail. With regard to mosquito-
proof buildings, even the signal-boxes on the railway are mosquito-
proof, and special arrangements have been made so that mosquitos
i35
cannot gain an entry where the wires run into these buildings to the
operating levers. The quarters in which the European labourers
live are mosquito-proof, but it is found that they are careless in
leaving the doors open at times. To safeguard them, mosquito
catchers are sent into these dwellings every morning to catch the
mosquitos which have gained entry. Most of the insects are filled
with blood, having fed during the night. Many mosquitos which
might have become infected are therefore destroyed. Dr. Orenstein
(1912) has found that this procedure reduces the malaria rate. Oil
and larvicide are used so freely that it is very difficult to find any
trace of larvae in pools not yet drained or filled in or in ditches not
yet concreted. In order to emphasise further the thoroughness of
the sanitation on the Canal Zone, I give the following list of the
personnel under Colonel Gorgas.
The personnel of the Division of Zone Sanitation is as
follows: —
1 Chief Sanitary Inspector.
1 Chief Assistant Sanitary Inspector.
2 Division Inspectors.
26 Inspectors.
1 Inspector—Entomologist.
18 Foremen and 226 labourers.
The following is a summary of the routine work accomplished
by this personnel during the month of July, 1912 : —
Anopheles work:
Linear feet of ditches cleaned.
Linear feet of new ditches dug.
Square feet of grass cut and removed.
Number of loads of grass removed to dump.
Number of cubic yards of earth used in filling
holes, etc.
Districts oiled, each four times in routine:
Chorrillo, Cocoa Grove and Avenida Ancon;
San Felipe, Santa Ana and Trinchera; Pueblo
Nuevo, San Miguel and Caledonia; Guachapali,
Maranon and Panama Railroad yards; special
oiling as found necessary.
Larvacide used .gallons
129,150
710
736,300
494
28
500
136
Disinfection and fumigation:
Houses disinfected and fumigated for diphtheria... 3
Houses disinfected and fumigated for typhoid fever 1
Houses fumigated for yellow fever. 6
Number houses disinfected . 4
Number houses fumigated . 10
Number rooms disinfected. 10
Number rooms fumigated . 38
Total number cubic feet disinfected . 16,300
Total number cubic feet fumigated. 369,300
Destruction of rats:
Rats caught in traps . 496
Number of rat traps in use daily . 200
Inspection of houses and yards:
Number of yards cleaned. 132
Loads of refuse hauled to dump. . 33
Number of old buildings condemned and
demolished . 1
Number of notices to abate nuisances served by
inspectors . 42
Number of inspections by reinspector . 123
Number of notices complied with . 39
Number of letters to alcade requesting enforcement
of sanitary rules and regulations . 1
New buildings :
Number of plans submitted to Health Officer and
approved . 19
The Sanitary Inspectors are thoroughly trained men. They
undergo courses of instruction and have to pass examinations.
I would refer those interested to the ‘ Manual of Instruction for
Sanitary Inspectors/ published by the I. C. C. Press, Mount Hope,
Canal Zone, 1912. Everything goes to show the thoroughness of
this great sanitary organisation.
ANTI-YELLOW FEVER WORK
Despite the fact that no case of yellow fever originated on the
Canal Zone since May, 1906, yet the efforts to eradicate the
Stegomyia fasciata have not been abated. The Sanitary Inspectors
137
exercise great vigilance in preventing accumulations of water in
containers where Stegomyia might breed. Roof gutters are not
allowed, except self-draining, short gutters over the entrance to the
house. No water containers are permitted at any house within a
300 foot radius of a public water supply, nor, of course, at any
house where there are water connections. In localities more than
300 feet distant from a public water supply, only screened
containers are allowed. The Sanitary Department supplies the
material for screening one container at each house. Measures for
eradicating Stegomyia have been so successful that an adult
Stegomyia is considered a great curiosity, and it is becoming
difficult to obtain Stegomyia larvae in the sanitated area of the
Canal Zone.
SANITARY MEASURES ADOPTED TO SAFEGUARD AGAINST
PLAGUE
The quarantine is exceedingly strict, as plague is endemic in
Guayaquil, which is only four days’ sail from Balboa. Houses are
rendered as far as possible rat proof by cementing the floors or by
building them on piles. Rats are constantly being trapped and
poisoned, and all rats caught or found dead are immediately
immersed in antiseptic and sent to the laboratory for examination.
Night soil and garbage are placed in special garbage tins with lids.
It is removed daily to be burnt and buried. It is the duty of the
Sanitary Inspectors to attend to this.
MEASURES AGAINST ANKYLOSTOMIASIS
This disease is very common, especially among the negroes.
It is combated by the establishment of a very thorough latrine
system along the line of the Canal. The Sanitary Inspectors look
after the latrines in their respective districts. The latrines are
fly-proof earth closets, and their position is frequently changed.
When the contents of the pit reach within two feet of the ground
surface, the pit must be filled, after liberally covering the contents
with unslaked lime or other disinfectant. The fill must be banked
at least 18 inches above the surrounding ground surface. Patients
coming into the hospitals and dispensaries are constantly examined
for ankylostome ova, and they are given a thorough course of
santonin treatment if these are found.
138
ANTI TYPHOID AND ANTI-DYSENTERY WORK AND GENERAL
SANITATION
The reservoirs and their watersheds are kept free from human
habitation and trespass, and the water is periodically examined
chemically and bacteriologically.
Garbage cans are cleaned daily, and the garbage is either burnt
or buried each day. Manure is burnt to prevent fly breeding. Pit
closets are rendered fly proof. Literature is distributed among
the people to call attention to the danger of flies.
Sanitary Inspectors carry out all the fumigation and disinfection
necessary. In bacterial diseases, such as diphtheria and typhoid
fever, thorough disinfection of utensils, floor clothing and bed
clothes is carried out.
THE HOSPITALS ON THE CANAL ZONE
There are two principal hospitals on the Canal Zone. One at
Colon on the Atlantic side of the Canal, with 450 beds, and the
other at Ancon on the Pacific side of the Canal, with 1,500 beds.
In addition there are several dispensaries along the line of the
Canal. These hospitals were in part located and built by the
French. They consist of a series of wooden buildings built on
piles and are entirely mosquito-proof (PI. XII, fig. 6). The buildings
are spread out after the fashion of a fever hospital in this
country, and the grounds are beautifully laid out. These
buildings, like all other American dwellings on the Canal Zone, are
for the most part two stories high. A few are three stories in height
and many only one storey. They are cool, airy and well shaded.
The wire-screening as a rule goes all round the building (fig. 6),
but in some cases, especially in the smaller buildings, only two or
three sides are screened. The remaining sides consist of wood, with
one or two screened windows. The outside walls, so to speak, are
therefore composed of wire screening in a wooden framework.
The roof consists of corrugated zinc or tile. The eaves, which
have no gutters except for the part immediately above the entrance
to the house, project about four feet beyond the screened frame¬
work, and the water from these falls into a concrete open ditch all
round the house. Inside the wire-screening is a space varying from
about three to four yards wide, forming a sort of protected
i39
verandah. Then comes the house proper with sitting-rooms and
bedrooms, or wards in the case of a hospital building. The walls
of the house proper consist of wood with doors and large lattice
windows, so that air or wind can pass through the whole building.
The rain practically never gets within the wire screen on account
of the projecting eaves, and the inhabitants live for the most part
in the verandahs immediately within the wire-screening, and
therefore practically in the open air. The doors forming the
entrance to the buildings are usually double, and consist of a
skeleton framework screened with wire. They have strong springs
attached so that they shut automatically. The screening consists of
pure copper gauze, eighteen meshes to the inch, and lasts as a rule
for three years or more.
The hospitals are as well equipped and up to date as any
European hospital. All the nurses are white, and each separate
building has a laboratory attached with up-to-date microscope and
other necessary equipment. The pathological department is
likewise large and up to date. At least two of the medical men
are employed chiefly on research work.
I feel sure that Ancon Hospital is the most modern and best-
equipped hospital in the whole of the tropics. There is only one
criticism, it seems to me, to be made regarding the anti-malaria work
on the Canal Zone at Panama, namely, that the malarial patients
are not subjected to a sufficiently long treatment with quinine.
Malarial patients (non-American) remain in hospital only for a few
days after their attack of fever has subsided under quinine treat¬
ment, and hence a relapse is almost certain. Many patients come
to hospital several times a year for malaria, and in the present state
of sanitation, it seems improbable that each of these attacks is
likely to be a fresh infection. In fact, I am inclined to believe that
probably 80 per cent, of the cases of malaria now occurring on the
Canal Zone are relapses due to an insufficient course of quinine
treatment. The mild character of the cases coming into hospital
and the number of re-admissions lead me to this belief. This fault
does not lie with the medical men, but is due to the fact that the
patients (non-American) do not receive their pay during their stay
in hospital, and therefore they are anxious to leave as soon as they
feel well, before being properly cured. I believe that a three weeks’
140
course of thorough quinine treatment safeguards against relapse in
most cases. If such a treatment could be given to all patients
before they were allowed to leave hospital, I feel sure that malarial
fever among the workers on the Canal would finally disappear.
The American patients, as a rule, receive a thorough course of
treatment before leaving hospital, and malaria is now practically
non-existent among them.
RESULTS OF THE SANITATION ON THE CANAL ZONE
In the introduction I have already mentioned the remarkable
improvement in health and comfort brought about since the
American occupation.
With regard to the remarkable statistical results of the sanitation
in Panama, I will again quote from Colonel Gorgas (1911), as
follows:— 4 The health conditions at Panama when the United
States took charge in 1904 were very bad. For four hundred years
this Isthmus had been considered the most unhealthy spot in the
world and the mortality records will sustain this opinion. . . .
At one time the construction company of the old Panama Railroad
imported 1,000 negroes from the West Coast of Africa, and
within six months these had all died off. At another time, for the
same reasons, they brought over 1,000 Chinamen and within six
months these had all died off. One of the stations at present on
the Panama Railroad is called Matachin. The tradition is that
this name is derived from the Spanish words mata, ‘ killed,’ and
chin, 'Chinamen/ because this was the point where the 1,000
Chinamen were housed and where most of them died. . . . The
French lost 22,189 labourers by death from 1881-1889. This
would give a rate of something over 240 per thousand per year.
. . . Our maximum rate in the early days was 40 per thousand;
our rate at present is 7 5 per thousand. . . . The malaria sick
rate we have reduced from 821 per thousand to 187 per thousand.
But most important of all, yellow fever has been entirely banished.
We have not had a single case since May, 1906. . . . The
sanitation has cost less than 1 per cent, of the total appropriation
for all purposes. . . . We hope that our success at Panama will
induce other tropical countries to try the same measures; and that
thereby gradually all the tropics will be redeemed and made a
suitable habitation for the white man. But if this is to come about,
it must be shown that it can be done at reasonable cost and within
the moderate means of the tropical region. ... I wish therefore
to say most emphatically that considering the results and
difficulties surrounding the subject, the sanitation of Panama has
not been costly. When the Canal shall have been finished it can
be shown that sanitation cost about 365,000 dollars (£73,000) per
year. For a population of 150,000 this means an expenditure of
about 1 cent (one halfpenny) per caput per day, and this sum
is well within the means of any tropical country.’
In another article (1910) he made the following interesting
statements:—‘But I do not believe that posterity will consider the
commercial and physical success of the Canal the greatest good
it has conferred upon mankind. I hope that as time passes our
descendants will see that the greatest good the construction of the
Canal has brought was the opportunity it gave for demonstrating
that the white man could live and work in the tropics, and maintain
his health at as high a point as he can, doing the same work, in the
temperate zone. That this has been demonstrated none can justly
gainsay. . . We therefore believe sanitary work on the Isthmus
will demonstrate to the world that the white man can live and work
in any part of the world and that the settling of the tropics by the
Caucasian will date from the completion of the Panama Canal.’
SANITATION IN TRINIDAD
On my return from Panama I was able to spend seven days in
Trinidad and four days in British Guiana, and although the time
was too short to look thoroughly into the question of sanitation in
these places, yet in that short period I was able to gain at least
some superficial information. My account of the sanitation in
these latter places is therefore much shorter than I would have
liked.
With regard to malaria in Trinidad, the report of the Surgeon-
General for 1911-1912 shows that this disease stands third in the list
as a cause of death. The number of deaths from malaria in that year
was 722. This is sufficient to show that malaria is very prevalent,
I 4 2
yet very little anti-malarial work is carried on there. One is
bitten by mosquitos frequently, yet there are practically no mosquito-
proof houses on the island except those of the American Asphalt
Company and some of the oil companies. Only one Government
residence has been made mosquito-proof, namely, that of the
District Medical Officer at Erin. A residence for the District
Medical Officer at La Brea is about to be Erected, but we believe
that the Finance Committee refused to vote the necessary funds for
screening it. Furthermore, neither funds nor encouragement are
forthcoming to eradicate even the smallest Anopheles breeding
places. Some oil and larvicide are employed in some breeding
places about the Port of Spain and at a few other places, but this
is only done spasmodically.
There are practically no men employed in anti-mosquito work
except in Port of Spain, where Stegomyia reduction against yellow
fever is persistently and more or less successfully pursued, but
much more might be done if more trained and efficient Sanitary
Inspectors were employed. There are five Assistant Sanitary
Inspectors in Port of Spain and three or four for the rest of the
colony, but how far these have been trained in tropical sanitation
I am unable to state.
With regard to anti-yellow fever work, there are no legal powers
to enforce the abolition of eaves gutters, which are an incessant
source of Stegomyia breeding.
Water barrels are rarely in evidence in the town because of the
ample pipe-borne water supply. This water supply, by the way,
is not above suspicion, as a considerable epidemic of typhoid fever
occurred last year. The streets in Port of Spain are beautifully
laid out and well paved, but there is no proper law with regard to
garbage disposal, and I understand that proper garbage receptacles,
with metal covers, are not provided by the General Board of Health.
Plague is endemic, and large sums of money are spent on rat
extermination, etc., yet all kinds of filthy house garbage are placed
in open boxes, old baths, barrels or baskets on the pavements, and
their contents are liable to be scattered about by fowls, dogs, etc.
There is much ankylostomiasis on the island, yet no estate
provides latrine accommodation for its coolie labourers, whether
indentured or not.
I need hardly point out, therefore, that in sanitation Trinidad
is far behind Panama. This is not the fault of the medical staff,
who are constantly urging the advisability of improvement in all
these matters. The fault lies with the municipalities and those in
power, who are slow to act or to give support either morally or
financially.
SANITATION IN BRITISH GUIANA
I have more praise for British Guiana, not because it is more
healthy than Trinidad, but because one can see more serious efforts
being made there with regard to sanitation than in the latter.
British Guiana is an exceptionally difficult place to deal with. All
the sea coast, including Georgetown the capital, lies below the
level of the sea at high water. A sea wall keeps out the sea during
high water, and numerous canals empty their contents through the
gates at low water. These canals are essential to drain off the
water which collects on this low-lying land. They cannot possibly
be dispensed with, but fortunately they are kept comparatively free
of mosquito larvae by the swarms of little fish which live in them.
Georgetown swarms with mosquitos, one cannot avoid being bitten.
It is a large town, beautifully laid out like a garden city, and most
of the mosquitos there have been bred in the town itself, or at
least very close to it. There are no mosquito-proof houses in the
town or elsewhere, and every house has rain gutters on the eaves,
leading into barrels or large tanks, as this is practically the sole
water supply for drinking purposes. These rain gutters are a
constant source of supply of mosquitos. Dr. Wise (1911) has
found that 58*3 per cent, of the premises in Georgetown are breeding
grounds for mosquitos, the great majority of these being Stegomyia
fascia/a. Several of the streets in the town also have old canals
running through them (PI. XIII, fig. 10). These are a serious
defect on account of mosquito breeding. Several of them have
been filled in by town rubbish (fig. 11), and finally converted
into very fine avenues (fig. 12). Unfortunately, however, the
amount of material available for filling in is small, so that these
old canals are only being filled slowly, and there are many of
them. The land around for miles is so flat that if material were
dug out for filling these canals, it would simply mean the
! 44
formation of other pools where the material was obtained. With
regard to the sugar estates, which employ thousands of coolies, the
anti-mosquito problem is even more difficult. The only alternative
is prophylactic quinine. In British Guiana, therefore, a decided
effort is being made to counteract malaria in spite of the difficulties,
but they fall far behind Panama in these efforts.
The Americans cleaned up their towns in Panama thoroughly
and at once. It would be wisest in the end to do the same in
Georgetown. The old canals in the town ought to be filled in
without delay. Material cannot be obtained from the land, but
it could easily be obtained from the muddy sandy river mouth by
means of quite a small suction dredger. A single steam shovel
could also do marvels in the soft land, in making further drainage
canals in the surrounding country if necessary. During my short
visit the floods were very extensive, and it was quite evident that
the canals could not cope with the rainfall.
Concerning the water supply and sewage system, they should
keep the example of Colon before them. It seems to me that in
the long run it would pay to lay down a pipe-borne water supply,
even though it had to be brought a long way. After all, water
tanks and rain gutters are a makeshift system and must give way
to a better some day. The sewage system is a very difficult
problem, yet it was solved in Colon by raising the level of the town.
In Georgetown I am glad to be able to state that the streets
are well paved, and that even in heavy rain there are no puddles.
Cement open gutters run all over the town. The money for this
purpose had to be borrowed, but the improvement brought about
is immense. There is not a single mosquito-proof house in
the town, but water barrels and tanks are carefully proofed
so that mosquitos cannot breed in them. This is excellent, but
there are still numerous cans, bottles and other rubbish, suitable as
breeding places for mosquitos, lying about in yards, especially
about the native dwellings. This should be immediately remedied.
With regard to the sugar estates, all water supply barrels are
mosquito-proofed (fig. 13), and ankylostomiasis is counteracted by
a good latrine system (fig. 14). In addition, a very large amount of
quinine is given as a prophylaxis against malaria to the coolies.
The result of this has been to reduce the malaria rate to about one-
*45
third of what it was originally. A good cold storage building has
been constructed, steps have been taken to improve the abattoir, and
a trained sanitary inspector has recently been obtained for the town.
Good work is therefore being done, but the great example of
Panama shows that it is possible to do much more, and more should
be attempted. For further details of the work being done in
British Guiana see papers by Dr. K. S. Wise (1911), Dr. E. P.
Minett (1912), and Wise and Minett (1912); also the reports of
the Surgeon-General (1912). The majority of the medical men in
both of these colonies are thoroughly progressive, but unfortunately
their progressive ideas are not always duly appreciated. The
sanitary organisation in these colonies cannot be compared with
the extraordinarily thorough system in Panama. I have stated
that there are a few doubtfully trained sanitary inspectors in
Trinidad, and only one recently appointed in British Guiana. The
medical men have, in consequence, to do practically everything
single-handed. It is extraordinary that, alone, they have been able
to do so much. The medical men unaided cannot organise such
a system as that in Panama. That is a matter which requires the
aid of the community and those in authority. Colonel Gorgas
has stated emphatically that his system has paid financially and
that, furthermore, the money required is well within the means of
any tropical country.
ments in order to catch the rain from the gutterless eaves of the
houses. These were the drastic measures enforced on the
inhabitants of Colon and Panama. The Americans lent the money
and supplied the labour. It cost the two towns about £455,000.
They are paying it back with interest through water rates which are
collected directly by the United States. The inhabitants who had
lived there in a slovenly way without interference, as their
forebears had done for centuries, were naturally very angry at this
apparent tyranny, but already they are becoming satisfied. Those
who have families have already noticed an unprecedented freedom
from disease among their children. Plague, typhoid fever,
dysentery and malaria are no longer to be feared, and now they
realise that what was done was good. The sanitation of Colon and
Panama is to-day as up-to-date as in any town in Europe. The
water-supply is examined very frequently by expert bacteriologists.
The quarantine at Panama and Colon is most rigid. Guayaquil in
Ecuador is an endemic yellow fever centre, and is only about four
days’ sail from Panama, hence a very strict quarantine is necessary.
In fact, if the Ecuadorians do not soon take steps to make their
town sanitary, it is highly probable that the Americans will force
them to do it, as it is a constant menace to the health of Panama.
Trinidad and all Venezuelan and Colombian ports are under
quarantine for yellow fever, though I believe that since my return
Trinidad has been removed from the list.
SANITATION ON THE ZONE OF THE CANAL-ANTI-MALARIA
WORK
On the Canal Zone the most prevalent disease has been malaria,
and much of it is pernicious malaria. The sanitary measures
adopted were, therefore, principally anti-mosquito measures.
Countless swamps and Anopheles breeding places existed along the
line of the Canal. A huge swamp lay behind Colon, and extensive
swamps also surrounded the town of Panama. After enormous
labour, these have been filled up by material excavated from the
Canal. Two methods have been adopted in filling the swamps.
One consists in dumping material (rock and earth) into them from
railway waggons. The other method, known as a ‘ hydraulic fill ’
(PI. XII, fig. 8), is to force into them a liquid stream of mud
and sand through pipes leading from large suction dredgers
employed in deepening the Canal. In many cases these pipes are
laid long distances, even a mile or more distant from the Canal.
I have seen swamps filled in this way to the depth of many feet,
so that only the tops of the trees which grew in these swamps are
now visible (fig. 8). Large unhealthy areas have been obliterated
in this way. The present town of Balboa on the Pacific entrance
to the Canal is being built on one of these filled-in swamps.
Strictly speaking, however, these huge undertakings can hardly be
considered as anti-malaria work. In describing this, I cannot do
better than give an abstract of a paper by Colonel Gorgas (1909)
on the subject.
He states that the anti-malaria work on the Isthmian Canal
cannot be well understood unless preceded by a brief description of
the sanitary organisation as it bears upon this work. The bulk of
the activities of the Department of Sanitation on the Isthmus has
no immediate bearing upon sanitation, namely, that which deals
with religious instruction, care of the sick, care of the insane,
of lepers, street cleaning, garbage collection, etc. The Department
of Sanitation spent in 1908 some two million dollars, but only
five hundred thousand dollars (£100,000) of this was spent on pure
sanitation. The American Canal Zone consists of a strip of land
ten miles wide, of which the Canal is the centre. This strip extends
about forty-five miles in length from North to South. The
population to be protected against malaria consists of about 50,000
labourers and their families, and is scattered all over this
five hundred square miles, though they are principally collected
along the line of the Canal, more particularly into some forty camps
and villages near this line. The temperature, rainfall, and
character of the terrain are all excellently suited for the breeding
of Anophelines all over this territory (fig. 7). The rainfall
averages over 100 inches yearly, and though there are four months
in which there is practically no rainfall, there is enough water for
the Anophelines to breed freely during these four months. During
the five years of occupancy of the Isthmus up till 1909, 250,000
people have resided in this zone, and these have been located
principally in places formerly unoccupied along the line of the
Canal, and as the villages are intended for only temporary
132
occupancy, the conditions are a good deal like those of an army
going into a new country, and increase the difficulty of sanitation
enormously.
The anti-malarial measures consist: —
1. In destroying the habitat of the Anophelines, during their
larval stage, within a hundred yards of dwellings.
2. Destroying within the same area all protection for the adult
mosquito.
3. Screening all habitations so that mosquitos cannot have
access.
4. Where breeding places cannot be destroyed by draining, use
is made of crude oil and larvicide.
These measures are given in the order in which they are
considered important.
For the purpose of carrying into effect these measures, the
five hundred square miles of territory have been divided into
seventeen districts. These districts are under the charge of a
Chief Sanitary Inspector, who has in his office the necessary clerical
force and three assistants. One of these assistants is especially
competent in the life-history and habits of the mosquito, another
in knowledge of ditching, tile-draining, etc., and the third in
knowledge of general executive work. Each of the seventeen
districts has a District Sanitary Inspector in charge. Each District
Inspector has a sufficient force of labourers (forty to fifty) to do the
necessary ditching and draining. A force of carpenters is required
to keep the screening in repair, and there are one or two quinine
dispensers who are kept constantly going around giving doses of
quinine to those who desire it. It is not attempted to enforce
prophylactic quinine. The Chief Sanitary Assistants are kept
constantly going over the work advising and instructing the District
Inspectors. The District Physician sends in daily to the central
office a report of the number of cases of malaria and the number of
the employees from which these cases come. This report is made
up weekly in the central office. A copy is sent to each District
Inspector, and he is held responsible for any excess of malaria in
his district. If the admission rate for malaria during the week
rises above per cent, something is considered wrong, and the
assistants of the Chief Sanitary Inspector are sent to look over the
133
ground to try to discover the cause. The District Inspector, for the
purpose of doing away with the breeding places of larvae, puts
down tile drains wherever they would be suitable. This is
considered the most effective and economical form of drainage.
After it is laid down it requires no more attention. There is no
breeding place left for mosquitos, as no water whatever is exposed
at the surface. A horse mower or scythe can be used for cutting the
grass over it. Where tile-drainage cannot be used, open concrete
ditches are put down. The first cost of this is nearly as great as
that of tiling, and a certain amount of labour is necessary to keep
the ditches clear. They must be swept out once a week. If the
ground cannot be drained in either of the above ways, open ditches
are used. This is the least effective and most expensive form of
drainage. In Panama they fill up rapidly with grass, and have
to be cleaned out about once in two weeks. They are always
breeding places for mosquitos, and have to be constantly treated
with oil or larvicide.
For the purpose of doing away with places which will harbour
the adult mosquito, the inspector clears the ground of brush and
grass for a hundred yards around the place to be protected. Where
the locality is to be occupied for a year or more, it is more
economical to grade the ground and plant grass, which can then be
kept down with a horse mower or scythe. A limited amount of
shrubbery and a few trees about a dwelling are not objectionable.
The inspector keeps the wire screening in repair by constantly going
over it with a force of carpenters. Good wire should last on the
Isthmus at least three years. The inspector uses crude petroleum or
larvicide in such places as cannot be drained, and in temporary
pools caused by construction or at temporary camps where it would
not be economical to drain. Usually when a new location is
occupied the malaria rate is high, frequently as high as 25 per cent,
a week, but always in the course of a month or two, when the
ground is drained and the brush cut, this drops to a
rate somewhere about 1 per cent. The above methods could be
applied to a considerable extent to military organisations. Where
troops remained at one camp for a week or longer, it would be
practicable to clear and drain the ground. It is most important that
the Sanitary Inspector should attend to the work himself. The men
•34
who do the ditching and brush-cutting, etc., should be immediately
under his control, and he should be held responsible for the proper
performance of the work, or in other words, the work must be
constantly supervised by a man who has a considerable knowledge
of these anti-malarial measures. Prophylactic quinine is looked
upon as an important measure, and is offered in three grain doses
to all employees who will take it. It is placed cm the table in all
the messes in liquid and pill form, and one to three dispensers are
employed in each district who go around the various villages.
Fuller details may be had from articles by the following
authors:—W. C. Gorgas(i904, 1906, 1907, 1908, 1909, 1910, 1912),
A. J. Orenstein (1911, 1912), J. A. Le Prince (1908), and
S. T. Darling (1910, 1912); also from the Reports of the
Department of Sanitation of the Isthmian Canal Commission by
W. C. Gorgas, and from the Canal Record.
The anti-malarial work, therefore, on the Canal Zone consists
simply in carrying out thoroughly and in a straightforward manner
the anti-malarial principles laid down by Ross soon after his
discovery of the transmission of malaria by the Anopheles mosquito.
Yet simple though these principles seem, the methods and
organisation of Colonel Gorgas deserve the most careful study. His
system has produced most remarkable results in a country which
was exceedingly unhealthy and in which the difficulties were great
and many, and although the construction of the Canal brought
unlimited labour and material to his aid, yet it also greatly increased
his difficulties, because it is well known that malaria tends to
increase greatly where land excavation is going on, on account of
the innumerable new breeding pools which are produced by these
excavations. This alone necessitates the constant inspection and
tackling of ever-changing conditions. The working population is
constantly changing its location, and this also involves many
difficult sanitary problems and increase of sanitary work. The
working population on the Canal moves about so much that these
methods are even suitable, as Colonel Gorgas has said, for military
organisations in the tropics. It seems to me that the keynote of the
success is thoroughness in every detail. With regard to mosquito-
proof buildings, even the signal-boxes on the railway are mosquito-
proof, and special arrangements have been made so that mosquitos
cannot gain an entry where the wires run into these buildings to the
operating levers. The quarters in which the European labourers
live are mosquito-proof, but it is found that they are careless in
leaving the doors open at times. To safeguard them, mosquito
catchers are sent into these dwellings every morning to catch the
mosquitos which have gained entry. Most of the insects are filled
with blood, having fed during the night. Many mosquitos which
might have become infected are therefore destroyed. Dr. Orenstein
(1912) has found that this procedure reduces the malaria rate. Oil
and larvicide are used so freely that it is very difficult to find any
trace of larvae in pools not yet drained or filled in or in ditches not
yet concreted. In order to emphasise further the thoroughness of
the sanitation on the Canal Zone, I give the following list of the
personnel under Colonel Gorgas.
The personnel of the Division of Zone Sanitation is as
follows: —
1 Chief Sanitary Inspector.
1 Chief Assistant Sanitary Inspector.
2 Division Inspectors.
26 Inspectors.
1 Inspector—Entomologist.
18 Foremen and 226 labourers.
The following is a summary of the routine work accomplished
by this personnel during the month of July, 1912 : —
Anopheles work:
Linear feet of ditches cleaned.
Linear feet of new ditches dug.
Square feet of grass cut and removed.
Number of loads of grass removed to dump.
Number of cubic yards of earth used in filling
holes, etc.
Districts oiled, each four times in routine:
Chorrillo, Cocoa Grove and Avenida Ancon;
San Felipe, Santa Ana and Trinchera; Pueblo
Nuevo, San Miguel and Caledonia; Guachapali,
Maranon and Panama Railroad yards; special
oiling as found necessary.
Larvacide used .gallons
129,150
710
736,300
494
28
500
136
Disinfection and fumigation:
Houses disinfected and fumigated for diphtheria... 3
Houses disinfected and fumigated for typhoid fever 1
Houses fumigated for yellow fever. 6
Number houses disinfected . 4
Number houses fumigated .
Number rooms disinfected. 10
Number rooms fumigated . 3 ^
Total number cubic feet disinfected . 16,300
Total number cubic feet fumigated. 369,300
Destruction of rats :
Rats caught in traps . 496
Number of rat traps in use daily . 200
Inspection of houses and yards:
Number of yards cleaned. 13’
Loads of refuse hauled to dump. . 33
Number of old buildings condemned and
demolished . 1
Number of notices to abate nuisances served by
inspectors . 4 2
Number of inspections by reinspector . 123
Number of notices complied with . 39
Number of letters to alcade requesting enforcement
of sanitary rules and regulations . 1
New buildings:
Number of plans submitted to Health Officer and
approved . 19
The Sanitary Inspectors are thoroughly trained men. They
undergo courses of instruction and have to pass examinations.
I would refer those interested to the 4 Manual of Instruction for
Sanitary Inspectors/ published by the I. C. C. Press, Mount Hope,
Canal Zone, 1912. Everything goes to show the thoroughness of
this great sanitary organisation.
ANTI-YELLOW FEVER WORK
Despite the fact that no case of yellow fever originated on the
Canal Zone since May, 1906, yet the efforts to eradicate the
Stegomyia fasciata have not been abated. The Sanitary Inspectors
137
exercise great vigilance in preventing accumulations of water in
containers where Stegotnyia might breed. Roof gutters are not
allowed, except self-draining, short gutters over the entrance to the
house. No water containers are permitted at any house within a
300 foot radius of a public water supply, nor, of course, at any
house where there are water connections. In localities more than
300 feet distant from a public water supply, only screened
containers are allowed. The Sanitary Department supplies the
material for screening one container at each house. Measures for
eradicating Stegomyia have been so successful that an adult
S/egomyia is considered a great curiosity, and it is becoming
difficult to obtain Stegomyia larvae in the sanitated area of the
Canal Zone.
SANITARY MEASURES ADOPTED TO SAFEGUARD AGAINST
PLAGUE
The quarantine is exceedingly strict, as plague is endemic in
Guayaquil, which is only four days’ sail from Balboa. Houses are
rendered as far as possible rat proof by cementing the floors or by
building them on piles. Rats are constantly being trapped and
poisoned, and all rats caught or found dead are immediately
immersed in antiseptic and sent to the laboratory for examination.
Night soil and garbage are placed in special garbage tins with lids.
It is removed daily to be burnt and buried. It is the duty of the
Sanitary Inspectors to attend to this.
MEASURES AGAINST ANKYLOSTOMIASIS
This disease is very common, especially among the negroes.
It is combated by the establishment of a very thorough latrine
system along the line of the Canal. The Sanitary Inspectors look
after the latrines in their respective districts. The latrines arc
fly-proof earth closets, and their position is frequently changed.
When the contents of the pit reach within two feet of the ground
surface, the pit must be filled, after liberally covering the contents
with unslaked lime or other disinfectant. The fill must be banked
at least 18 inches above the surrounding ground surface. Patients
coming into the hospitals and dispensaries are constantly examined
for ankylostome ova, and they are given a thorough course of
santonin treatment if these are found.
138
ANTI TYPHOID AND ANTI-DYSENTERY WORK AND GENERAL
SANITATION
The reservoirs and their watersheds are kept free from human
habitation and trespass, and the water is periodically examined
chemically and bacteriologically.
Garbage cans are cleaned daily, and the garbage is either burnt
or buried each day. Manure is burnt to prevent fly breeding. Pit
closets are rendered fly proof. Literature is distributed among
the people to call attention to the danger of flies.
Sanitary Inspectors carry out all the fumigation and disinfection
necessary. In bacterial diseases, such as diphtheria and typhoid
fever, thorough disinfection of utensils, floor clothing and bed
clothes is carried out.
THE HOSPITALS ON THE CANAL ZONE
There are two principal hospitals on the Canal Zone. One at
Colon on the Atlantic side of the Canal, with 450 beds, and the
other at Ancon on the Pacific side of the Canal, with 1,500 beds.
In addition there are several dispensaries along the line of the
Canal. These hospitals were in part located and built by the
French. They consist of a series of wooden buildings built on
piles and are entirely mosquito-proof (PI. XII, fig. 6). The buildings
are spread out after the fashion of a fever hospital in this
country, and the grounds are beautifully laid out. These
buildings, like all other American dwellings on the Canal Zone, are
for the most part two stories high. A few are three stories in height
and many only one storey. They are cool, airy and well shaded.
The wire-screening as a rule goes all round the building (fig. 6),
but in some cases, especially in the smaller buildings, only two or
three sides are screened. The remaining sides consist of wood, with
one or two screened windows. The outside walls, so to speak, are
therefore composed of wire screening in a wooden framework.
The roof consists of corrugated zinc or tile. The eaves, which
have no gutters except for the part immediately above the entrance
to the house, project about four feet beyond the screened frame¬
work, and the water from these falls into a concrete open ditch all
round the house. Inside the wire-screening is a space varying from
about three to four yards wide, forming a sort of protected
139
verandah. Then comes the house proper with sitting-rooms and
bedrooms, or wards in the case of a hospital building. The walls
of the house proper consist of wood with doors and large lattice
windows, so that air or wind can pass through the whole building.
The rain practically never gets within the wire screen on account
of the projecting eaves, and the inhabitants live for the most part
in the verandahs immediately within the wire-screening, and
therefore practically in the open air. The doors forming the
entrance to the buildings are usually double, and consist of a
skeleton framework screened with wire. They have strong springs
attached so that they shut automatically. The screening consists of
pure copper gauze, eighteen meshes to the inch, and lasts as a rule
for three years or more.
The hospitals are as well equipped and up to date as any
European hospital. All the nurses are white, and each separate
building has a laboratory attached with up-to-date microscope and
other necessary equipment. The pathological department is
likewise large and up to date. At least two of the medical men
are employed chiefly on research work.
I feel sure that Ancon Hospital is the most modern and best-
equipped hospital in the whole of the tropics. There is only one
criticism, it seems to me, to be made regarding the anti-malaria work
on the Canal Zone at Panama, namely, that the malarial patients
are not subjected to a sufficiently long treatment with quinine.
Malarial patients (non-American) remain in hospital only for a few
days after their attack of fever has subsided under quinine treat¬
ment, and hence a relapse is almost certain. Many patients come
to hospital several times a year for malaria, and in the present state
of sanitation, it seems improbable that each of these attacks is
likely to be a fresh infection. In fact, I am inclined to believe that
probably 80 per cent, of the cases of malaria now occurring on the
Canal Zone are relapses due to an insufficient course of quinine
treatment. The mild character of the cases coming into hospital
and the number of re-admissions lead me to this belief. This fault
does not lie with the medical men, but is due to the fact that the
patients (non-American) do not receive their pay during their stay
in hospital, and therefore they are anxious to leave as soon as they
feel well, before being properly cured. I believe that a three weeks'
140
course of thorough quinine treatment safeguards against relapse in
most cases. If such a treatment could be given to all patients
before they were allowed to leave hospital, I feel sure that malarial
fever among the workers on the Canal would finally disappear.
The American patients, as a rule, receive a thorough course of
treatment before leaving hospital, and malaria is now practically
non-existent among them.
RESULTS OF THE SANITATION ON THE CANAL ZONE
In the introduction I have already mentioned the remarkable
improvement in health and comfort brought about since the
American occupation.
With regard to the remarkable statistical results of the sanitation
in Panama, I will again quote from Colonel Gorgas (1911), as
follows:— 4 The health conditions at Panama when the United
States took charge in 1904 were very bad. For four hundred years
this Isthmus had been considered the most unhealthy spot in the
world and the mortality records will sustain this opinion. . . .
At one time the construction company of the old Panama Railroad
imported 1,000 negroes from the West Coast of Africa, and
within six months these had all died off. At another time, for the
same reasons, they brought over 1,000 Chinamen and within six
months these had all died off. One of the stations at present on
the Panama Railroad is called Matachin. The tradition is that
this name is derived from the Spanish words mata, ‘killed/ and
chin, ‘Chinamen/ because this was the point where the 1,000
Chinamen were housed and where most of them died. . . . The
French lost 22,189 labourers by death from 1881-1889. This
would give a rate of something over 240 per thousand per year.
. . . Our maximum rate in the early days was 40 per thousand;
our rate at present is 75 per thousand. . . . The malaria sick
rate we have reduced from 821 per thousand to 187 per thousand.
But most important of all, yellow fever has been entirely banished.
We have not had a single case since May, 1906. . . . The
sanitation has cost less than 1 per cent, of the total appropriation
for all purposes. . . . We hope that our success at Panama will
induce other tropical countries to try the same measures; and that
thereby gradually all the tropics will be redeemed and made a
suitable habitation for the white man. But if this is to come about,
it must be shown that it can be done at reasonable cost and within
the moderate means of the tropical region. ... I wish therefore
to say most emphatically that considering the results and
difficulties surrounding the subject, the sanitation of Panama has
not been costly. When the Canal shall have been finished it can
be shown that sanitation cost about 365,000 dollars 0673,000) per
year. For a population of 150,000 this means an expenditure of
about 1 cent (one halfpenny) per caput per day, and this sum
is well within the means of any tropical country/
In another article (1910) he made the following interesting
statements:—‘But I do not believe that posterity will consider the
commercial and physical success of the Canal the greatest good
it has conferred upon mankind. I hope that as time passes our
descendants will see that the greatest good the construction of the
Canal has brought was the opportunity it gave for demonstrating
that the white man could live and work in the tropics, and maintain
his health at as high a point as he can, doing the same work, in the
temperate zone. That this has been demonstrated none can justly
gainsay. . . We therefore believe sanitary work on the Isthmus
will demonstrate to the world that the white man can live and work
in any part of the world and that the settling of the tropics by the
Caucasian will date from the completion of the Panama Canal.’
SANITATION IN TRINIDAD
On my return from Panama I was able to spend seven days in
Trinidad and four days in British Guiana, and although the time
was too short to look thoroughly into the question of sanitation in
these places, yet in that short period I was able to gain at least
some superficial information. My account of the sanitation in
these latter places is therefore much shorter than I would have
liked.
With regard to malaria in Trinidad, the report of the Surgeon-
General for 1911-1912 shows that this disease stands third in the list
as a cause of death. The number of deaths from malaria in that year
was 722. This is sufficient to show that malaria is very prevalent,
1 + 2
yet very little anti-malarial work is carried on there. One is
bitten by mosquitos frequently, yet there are practically no mosquito-
proof houses on the island except those of the American Asphalt
Company and some of the oil companies. Only one Government
residence has been made mosquito-proof, namely, that of the
District Medical Officer at Erin. A residence for the District
Medical Officer at La Brea is about to be Erected, but we believe
that the Finance Committee refused to vote the necessary funds for
screening it. Furthermore, neither funds nor encouragement are
forthcoming to eradicate even the smallest Anopheles breeding
places. Some oil and larvicide are employed in some breeding
places about the Port of Spain and at a few other places, but this
is only done spasmodically.
There are practically no men employed in anti-mosquito work
except in Port of Spain, where Stegomyia reduction against yellow
fever is persistently and more or less successfully pursued, but
much more might be done if more trained and efficient Sanitary
Inspectors were employed. There are five Assistant Sanitary
Inspectors in Port of Spain and three or four for the rest of the
colony, but how far these have been trained in tropical sanitation
I am unable to state.
With regard to anti-yellow fever work, there are no legal powers
to enforce the abolition of eaves gutters, which are an incessant
source of Stegomyia breeding.
Water barrels are rarely in evidence in the town because of the
ample pipe-borne water supply. This water supply, by the way,
is not above suspicion, as a considerable epidemic of typhoid fever
occurred last year. The streets in Port of Spain are beautifully
laid out and well paved, but there is no proper law with regard to
garbage disposal, and I understand that proper garbage receptacles,
with metal covers, are not provided by the General Board of Health.
Plague is endemic, and large sums of money are spent on rat
extermination, etc., yet all kinds of filthy house garbage are placed
in open boxes, old baths, barrels or baskets on the pavements, and
their contents are liable to be scattered about by fowls, dogs, etc.
There is much ankylostomiasis on the island, yet no estate
provides latrine accommodation for its coolie labourers, whether
indentured or not.
I need hardly point out, therefore, that in sanitation Trinidad
is far behind Panama. This is not the fault of the medical staff,
who are constantly urging the advisability of improvement in all
these matters. The fault lies with the municipalities and those in
power, who are slow to act or to give support either morally or
financially.
SANITATION IN BRITISH GUIANA
I have more praise for British Guiana, not because it is more
healthy than Trinidad, but because one can see more serious efforts
being made there with regard to sanitation than in the latter.
British Guiana is an exceptionally difficult place to deal with. All
the sea coast, including Georgetown the capital, lies below the
level of the sea at high water. A sea wall keeps out the sea during
high water, and numerous canals empty their contents through the
gates at low water. These canals are essential to drain off the
water which collects on this low-lying land. They cannot possibly
be dispensed with, but fortunately they are kept comparatively free
of mosquito larvae by the swarms of little fish which live in them.
Georgetown swarms with mosquitos, one cannot avoid being bitten.
It is a large town, beautifully laid out like a garden city, and most
of the mosquitos there have been bred in the town itself, or at
least very close to it. There are no mosquito-proof houses in the
town or elsewhere, and every house has rain gutters on the eaves,
leading into barrels or large tanks, as this is practically the sole
water supply for drinking purposes. These rain gutters are a
constant source of supply of mosquitos. Dr. Wise (1911) has
found that 58*3 per cent, of the premises in Georgetown are breeding
grounds for mosquitos, the great majority of these being Stegomyia
fasciata. Several of the streets in the town also have old canals
running through them (PI. XIII, fig. 10). These are a serious
defect on account of mosquito breeding. Several of them have
been filled in by town rubbish (fig. n), and finally converted
into very fine avenues (fig. 12). Unfortunately, however, the
amount of material available for filling in is small, so that these
old canals are only being filled slowly, and there are many of
them. The land around for miles is so flat that if material were
dug out for filling these canals, it would simply mean the
! 44
formation of other pools where the material was obtained. With
regard to the sugar estates, which employ thousands of coolies, the
anti-mosquito problem is even more difficult. The only alternative
is prophylactic quinine. In British Guiana, therefore, a decided
effort is being made to counteract malaria in spite of the difficulties,
but they fall far behind Panama in these efforts.
The Americans cleaned up their towns in Panama thoroughly
and at once. It would be wisest in the end to do the same in
Georgetown. The old canals in the town ought to be filled in
without delay. Material cannot be obtained from the land, but
it could easily be obtained from the muddy sandy river mouth by
means of quite a small suction dredger. A single steam shovel
could also do marvels in the soft land, in making further drainage
canals in the surrounding country if necessary. During my short
visit the floods were very extensive, and it was quite evident that
the canals could not cope with the rainfall.
Concerning the water supply and sewage system, they should
keep the example of Colon before them. It seems to me that in
the long run it would pay to lay down a pipe-borne water supply,
even though it had to be brought a long way. After all, water
tanks and rain gutters are a makeshift system and must give way
to a better some day. The sewage system is a very difficult
problem, yet it was solved in Colon by raising the level of the town.
In Georgetown I am glad to be able to state that the streets
are well paved, and that even in heavy rain there are no puddles.
Cement open gutters run all over the town. The money for this
purpose had to be borrowed, but the improvement brought about
is immense. There is not a single mosquito-proof house in
the town, but water barrels and tanks are carefully proofed
so that mosquitos cannot breed in them. This is excellent, but
there are still numerous cans, bottles and other rubbish, suitable as
breeding places for mosquitos, lying about in yards, especially
about the native dwellings. This should be immediately remedied.
With regard to the sugar estates, all water supply barrels are
mosquito-proofed (fig. 13), and ankylostomiasis is counteracted by
a good latrine system (fig. 14). In addition, a very large amount of
quinine is given as a prophylaxis against malaria to the coolies.
The result of this has been to reduce the malaria rate to about one-
k
HS
third of what it was originally. A good cold storage building has
been constructed, steps have been taken to improve the abattoir, and
a trained sanitary inspector has recently been obtained for the town.
Good work is therefore being done, but the great example of
Panama shows that it is possible to do much more, and more should
be attempted. For further details of the work being done in
British Guiana see papers by Dr. K. S. Wise (1911), Dr. E. P.
Minett (1912), and Wise and Minett (1912); also the reports of
the Surgeon-General (1912). The majority of the medical men in
both of these colonies are thoroughly progressive, but unfortunately
their progressive ideas are not always duly appreciated. The
sanitary organisation in these colonies cannot be compared with
the extraordinarily thorough system in Panama. I have stated
that there are a few doubtfully trained sanitary inspectors in
Trinidad, and only one recently appointed in British Guiana. The
medical men have, in consequence, to do practically everything
single-handed. It is extraordinary that, alone, they have been able
to do so much. The medical men unaided cannot organise such
a system as that in Panama. That is a matter which requires the
aid of the community and those in authority. Colonel Gorgas
has stated emphatically that his system has paid financially and
that, furthermore, the money required is well within the means of
any tropical country.
146
REFERENCES
Darling, S. T. (1910). * Studies in Relation to Malaria,' Isthmian Canal Commission, Labora¬
tory of the Board of Health, Department of Sanitation, Gov. Printing Office, Washington,
1910.
- (1912). 'A Mosquito Larvacide-Disinfectant and the Methods of its Standardisation,’
American Jour, of Pub. Health, Feb., 1912.
Gorgas, W. C. (1904). ‘ Report on the Isthmian Canal,’ Engineering Record, New York City,
May, 1904.
- (1906). * Malaria in the Tropics,' Jour. Amer. Med. Assoc., May 5th, 1906.
- (1907). 1 Sanitary Work on the Isthmus of Panama during the last three years,' Medical
Record, May 18, 1907. Wm. Wood & Co., New York.
- (1908). 4 Method of the Spread of Yellow Fever,’ Medical Record, June 27, 1908.
- (1909). ' The Sanitary Organisation of the Isthmian Canal as it bears upon Antimalarial
work,' Jour, of the Assoc, of Military Surgeons of the United States, 1909.
- (1909). 'The Conquest of the Tropics for the White Race,’ Jour. Amer. Med. Assoc.,
June 19, 1909, Vol. LII, pp. 1967-1969.
- (19*0)- ‘The Expenses Necessary for Sanitation in the Tropics,’ Jour. South. Med.
Assoc., July, 1910, and Address of the President of the American Soc. of Trop. Med.
at the St. Louis Meeting, June 11, 1910.
- (*9*2)- ' Sanitation at Panama,’ Jour. Amer. Med. Assoc., March 30, 1912, Vol. LVIII,
pp. 907-909.
Le Prince, J. A. (1908). * Mosquito Destruction in the Tropics,’ Jour. Amer. Med. Assoc.,
Dec. 26, 1908, Vol. LI, pp. 2203-2208.
Minett, E. P. (1912). 4 Further Report on the Nastin Treatment of Leprosy carried out at
the Leper Asylum, Mahaica, British Guiana, from September, 1910, to September, 1911,’
Jour. London School of Trop. Med., Vol. I, Part iii, pp. 273-281.
Orenstein, A. J. (1911). ‘ Sanitary Inspection of the Canal Zone,’ Amer. Jour. Pub. Health,
March, 1912.
-(>9^2). 'Screening as an Anti-Malaria Measure,’ Engineering Record, June 29, 1912.
New York.
Wise, K. S. (1911). 'An Examination of the City of Georgetown, British Guiana, for the
Breeding Places of Mosquitos,* Annals Trop Med. and Parasit., Liverpool, Vol. V, No. 3,
Dec., 1911, pp. 435 - 441 -
Wise, K. S., and Minett, E. P. (1912). * Experiments with Crude Carbolic Acid as a Larvicide
in British Guiana,’ Annals Trop. Med. and Parasit., Liverpool, Vol. VI, No. 3, B,
Oct., 1912, pp. 327-330.
-(1912)* ‘ A Cheap and Simple Process for the Combined Clarification, Decolorisa-
tion, and Sterilisation of Peaty Waters,* Jour. Lond. School of Trop. Med., Vol. I,
Part iii, 1912, pp. 265-272.
-(1912). 4 Review of the Milk Question in British Guiana,’ Colonial Supplement,
Jour. Roy. Sanitary Instit., Oct., 1912, pp. 75-84.
OTHER LITERATURE
Reports of the Department of Sanitation of the Isthmian Canal Commission. Washington.
‘ The Canal Record,' Ancon, Canal Zone, Isthmus of Panama.
Report of the Surgeon-General, British Guiana, for the year 1911-1912.
Report of the Surgeon-General, British Guiana, dealing with Malarial and Anti-Malarial
Measures, 1912.
148
EXPLANATION OF PLATES
Plate XI
Fig. 1. Old drainage ditch. Colon.
Fig. 2. Street scene to-day, Colon.
Fig. 3. Early street scene, Colon.
Figs. 4 and 5. Thirteenth Street, Panama, before and after the
sanitary improvements. All the streets have been
renovated in the same manner.
Photos after Avery & Garrison , Panama .
Annals Trop. Med. & ParasitolVol. Vll
PLATE XI
150
Plate XII
Fig. 6. Mosquito-proof building (Nurses’ Home), Ancon
Hospital, Panama.
Fig. 7. Native labourers of sanitary staff clearing a swamp
preparatory to application of oil. Canal Zone,
Panama.
Fig. 8. ‘Hydraulic Fill,’ Canal Zone, Panama. Only tops of
trees visible. Sand and mud not yet dry.
Fig. 9. Ancon cemetery, Panama. Graves of French victims.
Thomson , photo , C. Tinline A*.. i.td.. /
* 5 *
Plate XIII
Fig. io. Old canal in Georgetown, British Guiana, not yet
filled in.
Fig. ii. Old canal, Georgetown, British Guiana, which has been
filled in with town rubbish.
Fig. 12. Old canal, Georgetown, British Guiana, which has been
filled in and transformed into an avenue.
Fig. 13. Coolie range, Diamond Sugar Plantation, Demerara,
showing water barrels which are all mosquito-proof.
Fig. 14. Latrines for the coolies at Diamond Sugar Plantation,
Demerara.
Annals Trop. Med. & Parasitol .. Vol. VII
>53
THE CULTIVATION OF ONE GENERA¬
TION OF BENIGN TERTIAN MALARIAL
PARASITES [PLASMODIUM VIVAX)
IN VITRO, BY BASS’S METHOD
BY
JOHN GORDON THOMSON, M.A., M.B., Ch.B.
(SIR EDWIN DURNING-LAWRENCE RESEARCH ASSISTANT)
AND
DAVID THOMSON, M.B., Ch.B., D.P.H.
(CLINICAL PATHOLOGICAL RESEARCH ASilSTAMT, SCHOOL OF TROPICAL MEDICINE, LIVERPOOL)
With Plate by H. B. FANTHAM, D.Sc., B.A.
(Received for publication 5 March, 1913)
Plate XIV
INTRODUCTION
In this Journal (Vol. VI, No. 4, Dec. 1912) J. G. Thomson
and S. W. McLellan described the cultivation of one generation
of Plasmodium falciparum. We are now able to give the following
details of the cultivation in vitro of one generation of Plasmodium
vivax. It may be mentioned that this parasite was cultivated up
to three-quarters growth by Dr. H. Carter and one of us (D. T.)
in Panama in December last.
CASE
J. F., male, aged 35, was admitted to the Royal Southern
Hospital, Liverpool, to Dr. Lloyd Roberts’s clinic, in February,
19 * 3 -
The patient had resided for some time in the Island of Java,
where he developed fever, with vomiting. On admission to hospital
a history of four weeks’ illness was given, and he complained of
rigors, fever, aching pains in the limbs and back, and severe
vomiting. Examination of his blood showed numerous young ring
*54
parasites. The temperature on admission was ioi° F. It dropped,
without quinine treatment, and on the third day another rigor
occurred, the temperature rising to 100*4° F. Ten c.c. of blood were
taken, while the temperature was at its height, for cultivation of
the parasites. Next day the fever had subsided, but on the
following day another severe rigor occurred, the temperature rising
to 104*6° F. Ten c.c. of blood were again taken from a vein, at the
height of the fever, for purposes of cultivation. Quinine was then
given in doses of ten grains thrice daily by the mouth, and no
further attacks of fever resulted, the patient making a good
recovery. The temperature chart is given below.
TECHNIQUE
As stated, 10 c.c. of blood were drawn, with aseptic precautions,
from the median basilic vein on two occasions, at the time when the
temperature was at its height, on the nth February and again on
the 13th February (see chart).
1 55
The blood on both occasions was transferred to a large sterile
test tube and defibrinated; it was then distributed into several
smaller test tubes and incubated at 39 0 C. The corpuscles settled
gradually, leaving about one half-inch of clear serum above them.
The amount of dextrose solution added was slightly more than
the amount recommended by Bass (1912).
EXAMINATION OF CULTURES
Experiment 1 . A blood smear, taken at the time of the with¬
drawal of the first 10 c.c. of blood on the nth February, showed
85 per cent, of young ring parasites and 15 per cent, of three-quarter
grown forms, also a few gametocytes. After nineteen hours’
incubation there were only 3 per cent, of young rings, 17 per cent,
were one-quarter grown, while 70 per cent, were one-half to three-
quarters grown, and 10 per cent, showed signs of presegmentation.
After twenty-three hours the majority of the parasites were three-
quarters grown.
No further examination was made till the forty-fifth hour of
incubation, and it was then found that 80 per cent, were small rings,
and 20 per cent, were one-half to three-quarters grown. Sporulation
had, therefore, occurred between the twenty-third and the forty-fifth
hours of incubation.
Experiment II. Blood smears taken at the time of the second
withdrawal of 10 c.c. of blood, on 13th February, showed 96 per
cent, of young rings, the remaining 4 per cent, of the parasites being
about one-half grown (PI. XIV, figs. 1-3). After eight hours’
incubation, 4 per cent, were young rings, 80 per cent, were one-
quarter grown, and the remaining 16 per cent, were three-quarters
to full-grown (figs. 4-10). After twenty hours, 65 per cent, were
one-half to three-quarters grown, with only 2 per cent, of young
rings, and a few sporulating forms (figs. 11-17). After twenty-nine
hours, 89 per cent, were young rings (fig. 18), 8 per cent, were
three-quarters grown, and again a few sporulating forms were
found. It is, therefore, evident that in this experiment the maximum
sporulation, which we again unfortunately missed, occurred between
the twentieth and twenty-ninth hours of incubation.
Although the blood was drawn during the height of the
temperature, that is, when the majority of the parasites would have
*56
been expected to be very young, yet they attained their full
development and sporulated in little more than half the time which
would have been thought requisite for this development.
This, however, is in accord with the results obtained in the
cultivation of P. falciparum by J. G. Thomson and McLellan
(1912). They found that sporulation occurred after twenty-six
hours of incubation. We do not think that the 96 per cent, of
young rings shown at the time the blood was drawn could have
been twenty hours old, so that it must be concluded that growth
took place more rapidly in the tube than it would have done in the
circulating blood of the patient.
MORPHOLOGY OP THE PARASITES IN CULTURES
The following remarks refer to Experiment II, and are illustrated
by the accompanying plate (PI. XIV). The morphology of the
benign tertian parasite (Plasmodium vivax ), as it occurs in the
peripheral circulation, is well known. Unlike the malignant parasite,
all stages may be found in the peripheral blood. In malignant tertian,
as a rule, only ring forms are found, as the further development of
the parasite takes place in the capillaries of the internal organs.
In the benign tertian several stages of the development are usually
found in a single smear of peripheral blood. Even the full-grown
parasites of benign tertian, which, by the way, are much larger than
the full-grown malignant Plasmodia , are not arrested in the fine
capillaries of the internal organs, but are apparently able to
circulate freely. A pure case of benign tertian malaria, in which
the parasites are all in the same stage of development, is very
rarely obtained, yet, as a rule, the majority of the Plasmodia are
found to be of the same age. In the cultivation of Plasmodium
vivax , therefore, if a full-grown parasite, or even a sporulating
form, is found, it must not be concluded that development has
taken place in the culture tube. In order to avoid being deceived
by this, we have made careful differential counts in the smears taken
at different periods of cultivation.
In our cultures the blood was drawn shortly after sporulation
of most of the parasites, so that the majority were young rings.
Figs. 1 and 2 show corpuscles with young rings, and already the
corpuscles contain many Schiiffner’s dots, and are somewhat
*57
enlarged, having a diameter of Qfi and io/ti respectively. In fig. 3
the corpuscle is 11*5/1 in diameter, and contains a parasite rather
more than one-quarter grown.
After eight hours’ incubation the blood was again examined, and
it was found that the parasites had increased in size in a very marked
degree; the majority were now one-quarter grown, and many were
three-quarters to full-grown. Fig. 5 shows a parasite about one-
quarter grown. Figs. 7 and 8 show half-grown parasites with
scattered pigment and the chromatin greatly increased in quantity.
Figs, g and 10 represent full-grown parasites, and show the
collection of pigment into a loose mass and marked increase of
chromatin.
In twenty hours the majority of the parasites were half, three-
quarters, or full-grown, and several were sporulating (figs. 11-17).
Fig. 15 shows the parasite with chromatin divided into seven
particles, or daughter portions. Figs. 16 and 17 show typical
sporulation, with production of 14 or 15 merozoites. In culture it
appears that the number of spores produced in benign tertian is less
than that produced in the malignant form of the parasite (Thomson
and McLellan, 1912). The maximum number of spores in
Plasmodium vivax averages about sixteen, but in some cases a few
more may be produced.
The culture was not again examined until the twenty-ninth hour,
and it was found that the majority of the parasites were then young
rings (fig. 18). There was, therefore, evidence that sporulation had
occurred and the young merozoites produced had entered new
corpuscles. No further asexual development took place.
DISCUSSION OF RESULTS
In the paper by J. G. Thomson and McLellan (1912) it
was noted that the parasites, after an incubation of twelve hours,
showed a definite increase in size, the pigment being collected
together into a circular compact mass. In twenty-four hours the
malignant tertian parasite was found to undergo segmentation. The
maximum number of merozoites counted was thirty, and it is quite
probable that thirty-two is the greatest number of merozoites
produced by Plasmodium falciparum after complete segmentation.
In their remarks, J. G. Thomson and McLellan pointed out the
i5«
great difference of opinion among various observers regarding the
number of spores produced by the segmentation of the malignant
tertian parasites, and reasons were given for this.
In some slides of smears of the spleen and bone marrow made
during autopsies of several cases of comatose malaria ( P . falci¬
parum ), sent by Dr. James from Panama, very numerous sporulating
parasites were found. The number of spores in these preparations
was counted by Sir Ronald Ross and one of us (D. T.). The
following table gives the number of spores found in one hundred
fully-developed malignant tertian parasites ( P . falciparum ) from a
spleen smear of a case of untreated comatose malaria, Panama : -
13
per
cent, contained
32
spores
2
> »
n )»
3i
t»
I I
»*
> t it
30
1 1
7
> >
» » if
29
1 f
25
t >
if » »
28
f t
3
>»
» » » »
27
t »
17
> f
it ft
26
11
2
tt
ft ft
25
t f
10
tt
H ft
24
f t
3
»»
ft a
23
11
5
» t
ft *t
22
y t
2
» t
ft tt
20
11
ioo ,, ,, ,, from 20 to 32 spores.
These numbers lead us to conclude that the maximum number
of spores, after complete segmentation, may be as high as thirty-
two. This is entirely in agreement with the findings in culture by
J. G. Thomson and McLellan, and is a higher number than has, up
till recently, been given. Some observers have put the number as
low as eight to ten (Stephens and Christophers, 1908).
Again, in a placenta smear from a case of comatose malaria,
P. falciparum , Panama, sporulating parasites were found which did
not fill the entire corpuscle. The average number of spores found
in these was only eight. The patient, however, had received
60 grains of quinine during the twenty-four hours preceding death,
so that the sporulation was either atypical due to the quinine, or
else the parasites had not reached their maximum development.
Dr. James, of Panama, also believes that those forms containing
few spores are not completely developed, or are atypical sporulating
forms due to the action of quinine.
*59
It may now be stated with confidence that the malignant tertian
parasite is capable of producing, under favourable conditions, a
maximum of thirty-two spores, as proved both by culture and by
examination of autopsy smears of cases of untreated comatose
malaria. Last year Sir Ronald Ross taught his students that the
maximum number of spores produced by the malignant tertian
parasite was 2 5 , while that of the benign tertian parasite was 2 4 .
Another point of great importance, noted in the cultivation of
P. falciparum by Thomson and McLellan (1912), was that there
was a great tendency to clumping of the parasites, both before and
after segmentation. This point is of great interest since we have
found no such tendency in the case of cultures of benign tertian.
This phenomenon explains why the malignant tertian parasites
accumulate in the capillaries of the internal organs after attaining
a certain size, only the corpuscles containing the young forms being
able to circulate in the peripheral blood. It would also explain the
blocking of the capillaries in the brain in the comatose form of the
disease.
In our cultivation of benign tertian, as already stated, no such
tendency to clumping of the parasites has been noted, and they
always remain well distributed throughout the smears. This
observation is in accordance with what we should expect, since in
the benign tertian there is little tendency to blocking of the
capillaries, with the resulting coma and other pernicious symptoms.
The corpuscles containing all stages in the development of this
parasite are able to circulate in the peripheral blood.
Another marked feature to be made out in cultivation is the
distribution of the pigment. In the figures given by J. G. Thomson
and McLellan for malignant tertian, it is clearly shown that the
pigment collects into a circular compact mass before segmentation
occurs, and remains so until the complete formation of merozoites.
This has also been seen in smears from the internal organs. In the
cultivation of P. vivax the pigment does not collect so early in the
growth of the parasite, and at no time does it appear to form so
dense or compact a mass as found in cultures of P. falciparum.
There was no evidence of haemolysis in the culture tubes.
i6o
POSSIBLE FORMATION OF GAMETOCYTES IN CULTURES
In a paper by one of us (D. Thomson, 1911), it was concluded
that crescents develop from the youngest forms of the asexual
parasite when a certain amount of immunity or resistance towards
the latter had developed in the human blood, and that the time
required for their development into an adult gametocyte was about
ten days. Reverting now to a consideration of the cultural
development of P. vivax, it was found that only one generation of
asexual parasites developed in the culture tubes. After the first
sporulation the parasites remained small, and did not pass through
another asexual phase. On the 5th day, however, it was noticed
in our cultures (P. vivax) that the parasites had changed much in
appearance, since all of them had become heavily pigmented and
compact (figs. 19-22), even the very youngest forms contained
pigment (fig. 19). There were no amoeboid straggling forms, all
were round and compact, and varied in size from very small circular
pigmented bodies to forms as large as 4/1 to 5 p in diameter (fig. 22).
The pigment was scattered more or less evenly throughout, and the
chromatin consisted of a single mass (fig. 21). The protoplasm had
a greyish-blue colour, quite unlike that of the asexual phase. On
the eighth day there was a very marked increase in the number of
large forms, varying from 4/1 to 6 n in diameter (figs. 23, 24). All
were circular, compact, and very heavily pigmented. On the tenth
day the cultures had dried up too much to permit of a further
examination.
We are doubtful as to the true explanation of these bodies.
They suggest gametocyte formation from their appearance, being
circular, compact, and heavily pigmented, and there were many
more large forms on the eighth day (figs. 23, 24) than on the fifth
day, which would appear to indicate that these bodies had been
growing in size. On the other hand, we must consider the
possibility that these are degenerating parasites, though it is
difficult to understand why degenerate parasites should develop such
a large amount of pigment, or why they should become compact and
show an apparent slow growth. We were unable to obtain any
flagellation in these bodies, and the corpuscles containing them
appeared to be shrunken.
SUMMARY
1. The benign tertian malarial parasite is capable of being
cultivated up to the stage of sporulation, for at least one
generation.
2. In our cultures the growth of the parasite from young rings
to sporulating forms took place more rapidly than in the blood of
the patient.
3. The cultures of benign tertian differed from those of
malignant tertian in that there was no tendency to clumping of the
parasites in the former, either before or during sporulation.
4. This difference appears to us to explain in a satisfactory
manner why only young forms of malignant tertian are found in
the peripheral blood, as the clumping tendency of the larger forms
causes them to be arrested in the finer capillaries of the internal
organs. It also explains the tendency to pernicious symptoms, such
as coma, in malignant tertian malaria. All stages of the benign
tertian parasite are found in the peripheral blood, and there are
seldom pernicious symptoms, because there is no tendency to
clumping.
5. Heavily pigmented, compact parasites were found in our
cultures of benign tertian on the fifth day. On the eighth day these
had grown larger, and their appearance suggested the development
of gametocytes.
6. The malignant tertian parasite ( P . falciparum ) is capable of
producing, in maximum segmentation, thirty-two spores. On the
other hand, benign tertian ( P . vivax) produces, as a rule, during
maximum segmentation, sixteen spores; sometimes more may be
produced, but the number is never thirty-two.
7. The pigment in P. falciparum collects into a definite,
circular, and very compact mass early in the growth of the parasite.
On the other hand, during the growth of P. vivax the pigment
remains scattered in definite granules throughout the body of the
parasite, till just before segmentation, when it collects into a loose
mass of granules in the centre of the full-grown Plasmodium.
ifo
REFERENCES
Bass, C. C. & Johns, F. M. (1912). The Cultivation of Malarial Plasmodia (Plasmodium vivax
and Plasmodium falciparum ) in vitro. Journ. Exp. Med., XVI, pp. 567-579.
Stxphxns, J. W. W. & Christoph tas, S. R. (1908). The Practical Study of Malaria, p. 34.
Thomson, D. (1911). A Research into the Production, Life, and Death of Crescents in
Malignant Tertian Malaria, in Treated and Untreated Cases, by an Enumerative
Method. Ann. Trop. Med. and Parasitol., V, pp. 57-82. 6 charts.
Thomson, J. G. & McLillan, S. W. (1912). The Cultivation of one Generation of Malarial
Parasites {Plasmodium falciparum ) in vitro, by Bass’s method. Ann. Trop. Med. and
Parasitol., VI, pp 449-462. 2 plates.
164
EXPLANATION OF PLATE XIV
The figures were drawn with an Abb€ camera lucida, using
ocular 4 and a Leitz i/i2th-inch oil immersion objective, from
Romanowsky stained preparations. The pigment is represented by
coarser dots in the protoplasm of the organism. The magnification
is 1,600 diameters, approximately.
Figs. 1 and 2 represent young plasmodia from the peripheral blood
before incubation, showing the corpuscles just beginning
to enlarge. SchiifFner's dots are well seen; no pigmenta¬
tion of the parasite is observed.
Fig. 3. Parasite is rather more than one-quarter grown. Corpuscle
gradually increasing in size. Chromatin increasing in
quantity. Schiiffner's dots present. Drawn immediately
before incubation.
Figs. 4-10 represent parasites after eight hours' incubation, showing
marked increase in size. Forms varying from small
rings to the full-grown parasite. Figs. 7 and 8
represent half-grown forms. Figs. 9 and 10 represent
practically full-grown forms. The parasites are now
distinctly pigmented, and in figs. 9 and 10 the pigment
is tending to collect in the centre of the parasite into a
loose mass. Note also the great increase in the amount
of chromatin.
Figs. 11-17 show parasites drawn after twenty hours’ incubation.
Figs. 15 to 17 show sporulating forms, the chromatin
being divided into fifteen daughter portions in fig. 17.
Fig. 18. Young parasite after twenty-nine hours' incubation.
Figs. 19-22. Highly pigmented small parasites in culture of five
days' incubation, suggesting the formation of young
gametocytes.
Figs. 23-24 show parasites from eight days' culture. These are
highly pigmented, and have increased in size. They are
probably young gametocytes.
FULMINATING CEREBRO-SPINAL
MENINGITIS IN JAMAICA'
BY
H. HAROLD SCOTT, M.D. Lond.
GOVERNMENT BACTERIOLOGIST
(Received for publication 7 March , 1913)
INTRODUCTION
Three short official reports have been already devoted to this
subject, namely: (1) On September 16th, 1912, in connection with
certain obscure cases of so-called 1 Vomiting Sickness 1 which
suddenly attacked a family in Franklin Town (Kingston) and
terminated fatally in four children within a space of twelve hours.
(2) On December 25th, 1912, when reporting upon the result of
an examination of cases of ‘ Vomiting Sickness * at Porus, in
Manchester.
(3) On January 7th, 1913; a further report on cases occurring
in the same district (Porus), where the disease had apparently taken
on an epidemic form.
Before entering on a description of the cases with which we
are essentially concerned, I should like to mention four cases (two
in April, one in June, and one in July, 1912) which were typical of
the ordinary sub-acute variety of Cerebro-spinal Meningitis, and
which, so to say, led up to the subsequent investigations. The
first was a female child of 12 months, and the duration of illness
was three weeks; the second was a male, aged 3 years, who lived
for 6 days; the third was a lad of 17 years of age, ailing for a
fortnight or so before the onset of any typical symptoms, and
death took place a fortnight later; the fourth, a woman of
19 years, who was ill for fully three weeks, possibly longer. This
also terminated fatally.
• This paper contains observations made up to Jan. 9th, 1913, at which date
Dr. Harald Seidelin, on expedition for the Liverpool School of Tropical Medicine, arrived
in Jamaica in order to investigate the so-caUed Vomiting Sickness. Dr. Seidelin suggests
that these observations should be published in order to make known the work which had
been done previously to his arrival.—H.H.S.
In all four a pure growth of meningococcus was obtained on
nasgar, and proved by sub-culturing on the same medium and on
ordinary agar, and incubating at 37 0 C. and at 25 0 C. Growth
occurred only at the higher temperature, and in no case on ordinary
agar until the third or fourth sub-culture. Maltose, dextrose and
galactose were fermented, saccharose, lactose and mannite were
unchanged.
SOME CASES IN KINGSTON
No similar cases came to my notice between July and September.
On the 4th of the latter month a sudden outbreak of cases of a
very acute nature occurred in Kingston itself, of which the following
is a brief history, as reported on the 16th.
A cigar-maker, named Adolphus Peart, his wife and seven
children, lived in a small house of two apartments, No. 6, Norfolk
Lane, Franklin Town. Three of the children, named Constantine,
Adolphus and Ruby, aged 8, 5 and 3J respectively, were a little
‘ out of sorts ’ and had slight attacks of vomiting during
Wednesday, September 4th, but were not, to all appearances,
sufficiently ill to call for any treatment, and at night the whole
family retired in ordinary health. About midnight, the youngest
(Ruby) began to vomit, and shortly afterwards Adolphus and
Constantine did the same. This, however, seems to have ceased
temporarily and the children fell asleep again. In the early
morning the second child (Adolphus) woke up, asked for some
food, but, before having any, was seized by a convulsive attack;
the other two (Constantine and Ruby) within a short period were
similarly convulsed, and all three lapsed into a state of coma.
A doctor was called in, but shortly after 7 a.m. the one first
attacked (Adolphus) died. The other two were brought to the
hospital at 9 a.m., suffered at intervals of a few seconds from
convulsive seizures, the spastic condition of the neck in the case
of the older patient, at all events, not completely relaxing even in
the intervals, and the younger (Ruby) died at 10 a.m., the older
(Constantine) at 10.55 a m., without recovery of consciousness in
either case.
In the meantime, the eldest girl of the family, Ethel, aged
14J years, started to vomit and within a few minutes became, to a
167
certain degree at least, unconscious. She was also brought to the
hospital, arriving at 9.30 a.m. When I saw her at 10.30, she
had recovered consciousness, but showed rigidity of the neck (and
possibly of the spinal muscles) and, while being examined, had a
convulsive attack; these were repeated, but were not so severe as in
the case of the boy. Nevertheless, the patient sank more deeply
into coma, and died at 5.55 p.m. Thus, four of the children had
died between 7 a.m. and 6 p.m.
No dietetic error could be discovered in any of the cases, and,
apparently, the only previous history obtainable was of a * cold in
the head’ for two or three days preceding. This, however, had
been disregarded as the condition is common in the city.
Post-mortem examinations were held in all four cases; the
details of three of them performed by me will be recorded shortly,
after the history has been completed.
At the funeral of these four on September 7th, another sister,
Violet, aged 11 years, suddenly complained of headache, vomited,
and became collapsed and semi-conscious. She, too, was hurried
off to the hospital. She had recovered consciousness by the time
she arrived, but showed a slight degree of rigidity of the neck
muscles; Kemig’s sign present on the right side, but less pronounced
or doubtful on the left. The vomiting was effortless, and not
accompanied by marked nausea; the pupils were equal and some¬
what dilated. This patient made a good recovery.
PATHOLOGICAL DETAILS
Cerebro-spinal fluid was taken by lumbar puncture from
Constantine, Ethel and Violet. (Ruby was dead in the hospital,
and Adolphus had died at home.) The fluid in the first two cases
was clear, in the last-named distinctly turbid. Enumeration of the
leucocytes in this last revealed 92 per cent, polymorphonuclar cells,
and in several were small Gram-negative diplococci. Cultures were
made from all three, and in every case the meningococcus was
isolated. The nature of the organism was proved by sub-cultivation
on various media and by fermentation tests. The same organism
was recovered from the intraventricular fluid of the child Ruby,
obtained post-mortem.
The macroscopic post-mortem signs were similar in every case,
but more marked in Ethel, who had been longest ill. There was
a distinct turbid haziness over the surface and base of the brain,
extending down the cord, with excess of fluid (cultivation of this also
yielded the organism), while in the case of Ethel the whole surface
of the convexity of the brain, and particularly the interpeduncular
space at the base, showed a much thicker layer of pearly lymph,
and here and there definite flakes of it.
These meningeal signs were common to all, varying only in
degree. The only other morbid conditions found were, in the case
of Constantine, some oedema of the lungs, and some scattered
petechiae on the surface (he had exhibited the most violent
convulsions), and in the case of Ethel there were firm adhesions
of the left pleura of old standing, and a similar but less marked
condition of the right. There were no signs of tuberculosis
anywhere in any of the cases.
The viscera of all four were sent in sealed jars to the Island
Chemist for examination, but he reported that no signs of any
poison were discovered.
The initial ' catarrhal* symptoms were in all probability what
has been noted as the 1 Premeningeal Catarrh,* set up by the presence
of the organism at its ‘ site of election * in the upper part of the
naso-pharynx, whence it spreads by way of the nerve canals through
the cribriform plate of the ethmoid to reach the meninges.
The mother, baby, and the son Reuben, were admitted to
hospital for observation, and swabs were taken from the upper
reaches of the naso-pharynx in each case. The meningococcus was
isolated from the boy’s throat, not from the other two. He was,
therefore, a ‘carrier* of the organism, and was detained at the
hospital.
The history of these patients is typical of nearly all the acute
cases of ‘Vomiting Sickness*; in fact, the medical man who sent
the patients to hospital stated that had they been attacked during
the winter months, he would have described them as typical
‘ vomiting sickness * cases.
This position is taken up by many physicians practising in
country districts, namely, that if they meet with cases in the colder
months they call them Vomiting Sickness, but at other times of the
169
year the deaths are certified as due to worms (which are present in
most of the native children), gastro-enteritis, food poisoning, etc.
In my opinion, this idea is responsible for many sporadic cases
being overlooked, and it is possible that such carry on the infection
through the summer and form the starting point of the localised
epidemics in the colder season.
From the cultures of the Peart cases I prepared a vaccine, not
so much with a view to curative treatment of subsequent cases,
for as a rule the course is so acute that no favourable result from
such treatment could be expected, death often occurring in
two or three hours after the onset of the first observed symptom,
but as a prophylactic in districts where the outbreaks mostly take
place.
Summary
1. In four cases presenting the typical symptoms of ‘ Vomiting
Sickness ’ the Weichselbaum’s diplococcus has been isolated.
2. These symptoms correspond to those associated with
foudroyante types of Cerebro-spinal Meningitis.
3. The organism was obtained from the cerebro-spinal fluid of
all those from whom this fluid was taken during life, and from
the cerebral ventricular fluid post mortem.
4. The organism was obtained from the naso-pharynx of one
of the contacts of these cases, though himself apparently in perfect
health.
5. The intracranial post-mortem signs were such as are
consistent with death from Cerebro-spinal Meningitis at such an
early stage, that is, with fulminating cases of the disease.
This focus did not spread, and the child (Reuben), who was
proved to be harbouring the organism in his naso-pharynx, did not
develop the disease.
METHODS OF INVESTIGATION
Having been struck by the peculiar hyperacuteness of these cases,
and by the remarkable similarity of the symptoms to those reported
from various districts of the island as occurring in the devastating
epidemics which arise every cold weather, preparations were made
170
for thoroughly testing the hypothesis that some at least of these
cases might be specially acute Cerebro-spinal Meningitis.
(< a ) Forms of report for filling in the histories of those patients
who were seen during life, and special forms for the
reporting of post-mortem findings were drawn up and
distributed to the various District Medical Officers.
(b) Considering that the meningococcus does not grow well on
ordinary nutrient agar, sloped tubes of nutrose ascitic agar
were prepared, placed in boxes containing also sterilised
swabs, slides, and ampoules of vaccine. These boxes
were likewise distributed to the District Medical Officers
with directions as to the use of the material contained.
The tubes of sloped nasgar were for inoculation with
cerebro-spinal fluid obtained by lumbar puncture, and for
inoculation with material obtained by means of the
sterile swab from the upper reaches of the naso-pharynx
of contacts. The slides were for smears of the spinal
fluid and of blood (as some of the cases might be
malarial). The vaccine was for use with contacts for
prophylactic purposes.
( c) Anti-meningococcal serum was ordered.
Since the organism, presuming that a variety of the meningo¬
coccus might be the cause, is far from being a resistant one, it was
considered that if the spinal fluid itself were sent, the organism
might die before the specimen reached the laboratory, so that
cultural tests could not be carried out. In some instances there is
an interval of over twenty-four hours between the time of despatch
of a specimen and its arrival at the laboratory. Secondly, that if
the organisms were present in small numbers only, they might be
overlooked in a smear. Thirdly, that if the inoculation of the
selective medium were made on the spot, colonies would in many
cases develop by the time the specimen reached the laboratory, and
examination of these and sub-cultivation could be made the same
day, and in this way a considerable saving of time would be
effected.
On the whole, I may say that this system has worked very well,
since in several instances the tubes on arrival at the laboratory
showed pure cultures, and in others there was sufficient growth to
enable plating to be carried out at once for purposes of isolation.
The procedure subsequent to the box and its contents reaching
the laboratory has been as follows: —
1. Smears were examined for enumeration of leucocytes and
the relative proportions in which they were present, and
for observations of any diplococci, intracellular or
otherwise. A second smear was stained by Gram's
method and counterstained. Sometimes, where nothing
definite could be made out from the smears sent of the
cerebro-spinal fluid, another smear of the liquid in the
culture tube would give indications of the varieties of
organisms present.
2. Any likely-looking colonies on the culture media were sub¬
cultured and examined.
3. The growth in the culture tube was plated for purposes of
isolation.
4. The colonies so isolated were then examined, and sub¬
cultures made on nasgar and ordinary agar and incubated
at 37 0 C. and at 25 0 C.
5. From the growth so obtained (only the former showed any,
in many instances) the effect on various sugar media was
noted. Dextrose, maltose, galactose, saccharose, mannite
and lactose were generally employed.
GENERAL SYMPTOMS
Returning to the description of the various cases with the
examination of which I have been concerned (prior to the
9th of January), it would be tedious, and would unduly prolong
this short paper, to give details of the history of each one. The
usual train of symptoms is as follows: —
The patient, usually a child, goes to bed apparently in its
ordinary health, or there may be a history of slight indisposition,
a cold in the head, or some loss of appetite, or a tendency to be
‘ droopy * and to lie down during the day preceding the actual onset.
During the night the child wakes up and vomits—perhaps only once,
perhaps three or four times—and complains of feeling ill. After
» 7 *
an hour or so he falls asleep again, and some three or four hours
later, more or less, again wakes up, complains of pain in the
abdomen (pain is used as a term for mere discomfort frequently
among the natives), and almost at once begins again to vomit,
usually frothy mucus, occasionally bile-stained, and later only
watery fluid, with, in most instances, little or no effort, unless the
stomach is quite empty, when troublesome retching ensues; if,
however, food or liquid is taken, there is apparently effortless
vomiting.
In a very short time, often a matter of a few minutes,
convulsions come on, and there is * stiffness of limbs and drawing
back of the head f (as the parents describe it), coma rapidly succeeds
and terminates in death.
In some there is no stiffness or retraction, but a general limp
condition.
The total duration is short, the average being some four to twelve
hours; the most rapid in my experience was thirty-five minutes.
Frequently, therefore, the patient is not seen during life, and
the history is both incomplete and unreliable, as it is obtained by
questioning the parents, who, not having noticed any details of
symptoms, will deny their presence, or affirm with equal readiness
in order to ingratiate themselves, for the acuteness of the course
formerly in many instances gave rise to the suspicion of poisoning.
When seen during life, the child is usually in the convulsive or
comatose stage. The temperature is rarely high, usually
ioi°-io2 °F., but it may be normal. The pulse-rate is between
90 and 100, fairly strong; respiration 26-30, regular till towards the
end, when the Cheyne-Stokes’ type may appear. Kemig’s sign is
present in some of the cases, and may be distinctly more marked
in one leg than in the other; rigidity of the neck muscles is more
common than retraction of the head, and this rigidity is often
overlooked because the flexion is not attempted in the strictly middle
line. Rigidity may be fairly marked, but when the flexion is
combined with lateral movement (as is the case where the test is
applied with the child lying on its side) the stiffness may be
masked, since lateral movement may be comparatively free in spite
of distinct rigidity of the neck muscles.
The pupils are usually equal, moderately dilated, and, if the
173
coma is not deep, react normally. In a few there is photophobia,
and in those retaining consciousness, general irritability and
complaint of headache—not always by any means severe—usually
frontal, sometimes general. Delirium is, so far as I have seen,
quite uncommon; shortly before passing into the comatose state, the
child may remark that it ‘ feels very bad,’ but does not call attention
to any particular symptom, or locate the pain, if such is complained
of, to any particular spot.
In cases which do not end fatally, the state of coma is rarely
present; there is vomiting, headache, convulsions with temporary
loss of consciousness only, and recovery is almost as rapid as the
onset. Within twenty-four hours a child, who has been seriously ill,
may be sitting up in bed, and in twenty-four to forty-eight hours
later be up and about, showing practically no symptoms except a
little pallor, general debility which rapidly clears up, and some
residual headache of no great severity, while others in the family,
who did not seem to be any worse at the time, have passed into a
state of coma and difed in a few hours.
FURTHER CASES
In the following descriptions, where the words ‘ usual history ’
are employed, the course of events has been that sketched above.
The next case, after the Franklin Town outbreak, which came
under my own personal notice (the tenth of the series of which
I have notes) occurred also in Kingston, on October 1st, 1912.
This, too, ran a more prolonged course than the generality of cases
here.
(10) Male child, aged 6 years, was apparently quite well on
September 25th. During the night he awoke and complained of
intense headache, calling out ‘ My head, my head.’ Went to sleep
again, but early in the morning of the 26th awoke again and
called out with headache, was seized with convulsions and lost
consciousness. Slight vomiting only, but much more severe next
day.
When admitted to hospital (October 1st) there was definite
retraction of the neck, Kernig’s sign marked, pupils dilated and
equal. Child quite conscious. Cerebro-spinal fluid turbid and
1 74
showing well-marked intracellular cocci, Gram-negative and diploid
in arrangement, with 84 per cent, of the leucocytes polymorpho¬
nuclear.
A good growth of meningococci was obtained, from which a
vaccine was made.
During the succeeding three weeks the patient's condition got
gradually worse; there was general flexion and irritability,
retraction of neck, wasting, etc., but very few twitchings and still
some vomiting. There was no serum at hand, so an injection of
100 cocci was administered on October 24th. Note made on the
26th:—‘Patient brighter and improving, temperature lower.' On
November 7th:—‘Improvement steadily maintained; retraction
gone, Kemig’s sign absent, is getting up and taking plenty of
nourishment.’ Two days later he went home, and reports state that
he is in good health but exhibits violent outbursts of temper on
trivial grounds, quite contrary to his former disposition.
This was in onset like the ordinary hyperacute Cerebro-spinal
Meningitis, but later took on the characters of the subacute and
recovered.
(11) P. McM., female, aged 4 years. History typical, namely,
apparently well at 11 p.m., October 12th; early on the 13th was
dull and restless (usually very 'bright and cheery), doubtful
headache—child held its head, but did not make any definite
complaint of it. About 7 a.m. child had a convulsion with the
head thrown back, and general twitching and stiffness. Rapidly
lapsed into coma and died without recovering consciousness.
At the autopsy the vessels of the surface of the brain were
congested; there was a pearly haze over the hemispheres, with flakes
of lymph in two or three spots. Ventricles contained excess of
fluid; smear of cerebro-spinal fluid showed intracellular Gram¬
negative diplococci. Culture obtained on nasgar, and proved by
subsequent tests to be the meningococcus.
(12) , (13), (14). Three children, sisters, aged 8 years, 2 years,
and 9 months, were brought to the hospital on October 23rd, 1912.
All three had a history of headache and vomiting, and Amy (the
one of 2 years) showed some staggering—she was old enough to
walk straight when well—stiffness of neck muscles was present in
all, Kernig’s sign in the two older, but doubtful in the baby.
i7S
Swabs taken from high up the naso-pharynx (lumbar puncture was
not permitted) gave meningococcus on cultivation, mixed with
Micrococcus catarrhalis and some Gram-positive bacilli (Hoffmann),
but separated by plating. They were all three seriously ill for the
next week or so, especially the two younger, but after that made
steady progress towards recovery.
It may be added that the mother of these children complained of
headache and of pain along the neck and shoulder, but the organism
was not obtained in her case.
(15) J. L. B., male, aged 4 years, October 19th. History
given was that which has been described at length above. The
duration of illness was only two hours. At the post-mortem
examination the ‘ meninges were markedly congested, fluid in excess
in the cerebral ventricles, and at the base of the brain. The fluid
was turbid uniformly.’ (Quoted from notes sent by Dr. Purchas.)
No specimens forwarded, so the cause was not proved.
(16) C. M., male, aged 13 years. Reported December 1st, 1912,
from Montego Bay. The only indication of anything which might
be regarded as a premonitory symptom was a ‘ fresh cold ’ for a
week preceding the sudden onset of vomiting at 4 p.m. The patient
became unconscious at 6 p.m., convulsions started at 7 p.m. and
recurred at intervals till death at 3 a.m. the following day. The
total duration was, therefore, 11 hours.
In this case there were a few ascarides found; the mesenteric
glands were enlarged. The organs, except the brain, were reported
as normal. But here the ‘ meninges were markedly congested,
especially over posterior halves of the hemispheres, and cerebellum,
and at the base. Fluid was in excess, turbid and milky.'
The meningococus was isolated from the culture sent.
(17) C. M. A., female, aged 4 years. December 7th. Usual
history, except that the patient was said to have retained conscious¬
ness till death. The medical attendant states that he did not see
the patient during life, and that the ‘ father’s statements as regards
the symptoms are unreliable.’ Duration of illness 15 hours.
Culture of spinal fluid yielded the meningococcus.
(18) H. W., female, aged 6 years. December 7th. Usual
history.—First symptom—vomiting—‘ started early in the morning,
and death occurred at 10 a.m.’ Smears of spinal fluid gave a
176
differential leucocyte count of 82 per cent, polymorphonuclear,
13 per cent, lymphocytes and 5 per cent, endothelial cells, and
showed Gram-negative diplococci mixed with coliform organisms.
Smears of intraventricular fluid gave 85 per cent, polymorphs,
12 per cent, lymphocytes and 3 per cent, endothelial cells, and the
same organisms.
The culture was badly contaminated, probably owing to the long
interval (fifty-six hours) between death and the autopsy, and I was
unable to isolate the diplococci. This case, therefore, was associated
with a Gram-negative diplococcus in the spinal fluid, the nature of
which was not definitely proved.
(19) A. J., male, aged 2 years. December 19th. Usual history
of vomiting, succeeded by coma, but there was no mention of
convulsions. Duration of illness six hours. Meninges dull, vessels
congested; fluid in ventricles and spinal canal in excess and turbid.
The fluid sent showed Gram-negative diplococci, but they did
not grow on the culture medium.
(20) R.T., female, aged 6 years. Porus District. December
22nd, 1912. This is a very good instance of the condition under
description, because, having been sent down to investigate a small
outbreak at Porus, I was able to see the patient myself during life
and to take specimens of the blood and spinal fluid, and to perform
the autopsy almost immediately after death took place.
The patient was a well-nourished child, and was said to have
been quite well until 6 p.m. on December 21st, when an attack of
vomiting came on. There was little or no complaint of headache.
After sleeping for several hours, she commenced to have attacks of
convulsions at intervals, from 4 a.m. on December 22nd, during
which she ‘went quite stiff.’ When I saw the patient at 12 noon
she was comatose. There was a general limp condition of the
muscles, but on flexing the head in the middle line the neck muscles
felt hard and stiff. Kernig’s sign was marked in both legs, but
more so in the left than in the right. There was a small group of
herpetiform spots on the lips at the right side of the mouth, but no
discernible rash on the body.
I took a blood smear and also spinal fluid smears and some of
the fluid for culture. There was no recovery of consciousness, and
l 77
death took place at 12.30 p.m.; the total duration from the first
onset of symptoms being i8£ hours.
All the organs appeared normal except that the mesenteric
glands were enlarged, and the brain and meninges were markedly
congested; there was a general haziness over convexity and base,
more over the right hemisphere, and in parts there was distinct
opalesence to milkiness. The ventricles were filled with clear fluid.
The smears of the spinal fluid taken showed many Gram¬
negative diplococci, several intracellular, but more lying free. The
differential count of the leucocytes gave polymorphonuclears
26 per cent., lymphocytes 67 per cent., endothelial 7 per cent. The
blood smear showed well-marked diplococci; this is the only case
in which I have seen them in a blood smear taken during life,
though it is only right to say that I have had very few opportunities
of examining blood smears taken shortly before death.
The culture tubes showed several small colonies of Gram¬
negative diplococci which subsequent tests proved to be meningo¬
cocci; and in this instance, therefore, not only was there definite
Cerebro-spinal Meningitis, but a condition of meningococcaemia was
shown to be present.
(21) D. D., male, aged n years. December 20th, 1912. The
history in this case was typical, except that the symptoms were a
little more drawn out. He complained of frontal headache and
chilliness on December 18th at 11 a.m., and vomited in all sixteen
times during that and the succeeding day. Was walking about in
the evening of the 19th, and complained about n p.m. that the
lamplight hurt his eyes. He was restless all that night, and had
frequent convulsive movements of the limbs with retraction of the
head and neck, and general signs of cerebral irritation till 4 a.m.
on the 20th, when coma supervened, and he died at 11 a.m. The
duration, therefore, was forty-eight hours.
Post-mortem report stated that there were some round worms
in the intestines and some congestion of the mucous membrane of
the stomach, but beyond this the only abnormality noticed was
marked congestion of the meninges, and a general dulling and loss
of glistening appearance of them, with excess of fluid in the
ventricles.
No cultures or specimens were sent from this case, so the proof
176
differential leucocyte count of 82 pci
13 per cent, lymphocytes and 5 per
showed Gram-negative diplococci mixt
Smears of intraventricular fluid gave
12 per cent, lymphocytes and 3 per cei
same organisms.
The culture was badly contaminates
interval (fifty-six hours) between death
unable to isolate the diplococci. This c.
with a Gram-negative diplococcus in t!
which was not definitely proved.
(19) A. J., male, aged 2 years. D<
of vomiting, succeeded by coma, bi
convulsions. Duration of illness six h
congested; fluid in ventricles and spin
The fluid sent showed Gram-nega'
not grow on the culture medium.
(20) R.T., female, aged 6 years.
22nd, 1912. This is a very good in
description, because, having been sen
outbreak at Porus, I was able to see
and to take specimens of the blood ai
the autopsy almost immediately after
The patient was a well-nourished
been quite well until 6 p.m. on De<
vomiting came on. There was littl
After sleeping for several hours, sh
convulsions at intervals, from 4
which she ‘went quite stiff.’ Wh
she was comatose. There was
muscles, but on flexing the head
felt hard and stiff. Kernig’s s
more so in the left than in the
herpetiform spots on the lips ;r
discernible rash on the body.
I took a blood smear and
the fluid for culture. Then
intestinal upset, or for the matter
• I with vomiting, but it may be
•is ted contacts have so far been
r J >r proved cases who had been
1 irom a boy of 7 years of age,
•se case the meningococcus was
• irochaete-like bodies, or long
■tther these have any causal
p>t say, but I may mention, in
h I have seen these bodies, the
bi'i. The second died in seven
filing. In neither case did this
edium used.
p rted with symptoms similar to
bid cultures were sent up from
Bigococcus was isolated; in three
g tube there was a slight growth
transit of the culture to the
ci the laboratory had been closed
was, in excess of zeal on the part
d the organism had been killed,
ier.
i:e treatment
il effects from vaccine treatment in
ft has been already stated. The
lactic use mainly, and secondarily
ic met with. For the acute and
lit^ococcic serum was ordered, but
| 1st its employment even in many of
t ulty of keeping the serum in warm
ce is obtainable; secondly, though it
T arts of the island where ice can be
f >f the cases are so rapidly fatal that
am arrives, although it may have been
It ice of the earliest symptoms and have
messenger without delay; thirdly, the
178
of the cause was not forthcoming. The case is described for two
reasons: firstly, because several similar cases followed it in which
the organism was detected; and secondly, because it is a good
example of the partial recovery or comparatively calm interval
between a violent onset and an equally violent return of more
serious symptoms which usually terminate fatally.
(22) R. E., male, aged 7 years. December 24th. Same district.
Usual history. 4 Duration 24-30 hours.’ Spinal fluid gave typical
cultures, and the meningococcus was proved.
The last-mentioned cases were the forerunners of a small
epidemic in this district. Details of the individual cases need not
be given, as they would be largely a repetition of what has been
already stated; nevertheless, there are some points of interest about
them, which may be briefly summed up as follows: —
Nineteen cases occurred during the last week of the year 1912,
of which notes were sent to me. In nine of these no lumbar puncture
was performed, and therefore, however typical the symptoms, they
cannot be said to be proved cases of Meningitis. In seven of the
nine, however, nearly all the classic symptoms—headache, vomiting,
irritability, rigidity, Kernig’s sign—were present, so that there is a
fairly strong probability that these were instances of the disease;
two of them recovered within three days and the rest within a week.
In six of the remaining twelve there was no symptom recorded
except vomiting; these rapidly cleared up with a stomachic mixture,
and I think they were probably merely gastric disturbances of a
mild type.
Lastly, in six of the ten specimens of fluid sent up, the meningo¬
coccus was isolated from the spinal fluid and proved; three of them
terminated fatally—one aged 7 years was ill for thirty hours; a
second, aged 3! years, died after ten hours’ illness; the duration in
the third, an adult, could not be ascertained.
Twenty-two subjects, nine of them contacts, and thirteen showing
what was thought to be early symptoms of the condition—vomiting
(with no ascertainable cause), headache and malaise—were given an
injection of the vaccine which had been prepared by me from a
former case. Of these thirteen, three were proved by culture from
the spinal fluid to be definite cases of Cerebro-spinal Meningitis; of
the remaining ten there is no evidence to show whether they were
i?9
cases of meningitis, worms, gastro-intestinal upset, or for the matter
of that any other disease associated with vomiting, but it may be
mentioned that none of the vaccinated contacts have so far been
attacked, and none of the suspected or proved cases who had been
vaccinated have died.
In the smear of the spinal fluid from a boy of 7 years of age,
who died in ten hours, and in whose case the meningococcus was
isolated, I found also several spirochaete-like bodies, or long
somewhat tapering bacilli. Whether these have any causal
connection with the disease I cannot say, but I may mention, in
passing, that in two cases in which I have seen these bodies, the
course has been exceptionally rapid. The second died in seven
hours after the onset of the vomiting. In neither case did this
peculiar organism develop on the medium used.
Of eleven cases subsequently reported with symptoms similar to
those already detailed, spinal fluid cultures were sent up from
seven. In three instances the meningococcus was isolated; in three
no growth occurred; in the remaining tube there was a slight growth
which had developed during the transit of the culture to the
laboratory, but on arrival there after the laboratory had been closed
for the day, the box containing it was, in excess of zeal on the part
of the recipient, placed on ice, and the organism had been killed,
for sub-cultivation failed altogether.
NOTE ON VACCINE TREATMENT
I did not expect any beneficial effects from vaccine treatment in
the acute cases of the disease, as has been already stated. The
vaccine was intended for prophylactic use mainly, and secondarily
for chronic cases should such be met with. For the acute and
hyperacute cases the anti-meningococcic serum was ordered, but
there are points militating against its employment even in many of
these. Firstly, there is the difficulty of keeping the serum in warm
climates in districts where no ice is obtainable; secondly, though it
has been stored in various parts of the island where ice can be
procured, nevertheless, many of the cases are so rapidly fatal that
death occurs before the serum arrives, although it may have been
telegraphed for on appearance of the earliest symptoms and have
been despatched by special messenger without delay; thirdly, the
i8o
serum must be injected intraspinally and repeated on three successive
days at least, and if the case terminates fatally, the parents are
almost certain to ascribe the death to the medical man’s action in
injecting fluid into the spinal canal.
Apart from the cases in which the vaccine was used prophy-
lactically, I do not think that the conclusion is justified that the
happy results following its employment in some of the cases referred
to above can be attributed to its use. The condition was too severe
for the small dose employed to have much effect, if any; also the
amelioration was too rapid and has been too well maintained to
justify any inference that the improvement leading to recovery was
the direct result of the small injections made. I feel that, though
‘ post hoc/ there are no grounds for regarding the recovery as
' propter hoc/ at all events until further reports of the efficacy of
the vaccine in similar cases have been received.
GENERAL SUMMARY
1. In several cases of illness with sudden onset in apparently
healthy subjects, terminating fatally in a high percentage of those
attacked, I have isolated a Gram-negative diplococcus from the
spinal fluid.
2. This organism gives the morphological and cultural
characters of Weichselbaum’s Diplococcus intracellular is meningi¬
tidis , except that in galactose it does not always give a definite
reaction, the medium in some instances not being affected. The
maltose and dextrose, however, are always typically acted upon.
3. These cases exhibit in many instances a symptom-complex
which has for years been spoken of in Jamaica as ‘ Vomiting
Sickness.’
4. The disease breaks out in localised epidemic form in various
parts of the island every cold weather, that is, from about the
middle of December to the end of February or beginning of March.
5. The disease occurs at other times as sporadic cases, but
apparently does not spread extensively.
6. It is a curious thing that practically all the cases are of the
hyperacute variety, either recovering in a few days, or dying in a
few hours. One rarely sees the subacute cases during the epidemic
times, and never, so far as I am aware, the chronic ones, unless
some of the natives one meets with who are chronically deaf, or have
seriously defective vision, or who suffer from fits, are instances of
Cerebro-spinal Meningitis with permanent sequelae. Of this there
is no proof.
7. In two instances a spirochaete-like body, or long curving
bacillus has been seen in the smears made directly from the fluid
obtained by lumbar puncture. What part, if any, this takes in
producing the symptoms I have had no means of discovering.
8. The disease is very rare amongst the white population;
I have known of only three instances so far during the past year.
Possibly this is due in part to better hygienic conditions, less
crowding, and so forth, but not entirely, because
9. I have not personally met with the condition once amongst
the East Indian population, where overcrowding and bad hygiene
are nearly, if not quite, as marked as in the case of the native.
For this, also, I am unable to offer any explanation at present,
though it may be noted that the food supply of the East Indian is
usually better than that of the West.
Pathological Laboratory,
The Public Hospital,
Kingston, Jamaica, Feb. 18, 1913.
Volume VII
June, 1913
No. 2.
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Editor
Professor Sir RONALD ROSS, K.C.B., F.R.S., Major I.M.S. (Ret.),
M.D., D.P.H., F.R.C.S., D.Sc., LL.D.
In Collaboration with
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
Professor J. L. TODD, B.A., M.D., C.M. McGill, D.Sc. Liv
H. WOLFERSTAN THOMAS, M.D., C.M. McGill.
ANTON BREINL, M.D.
H. B. FANTHAM, D.Sc.Lond., B.A. Cantab., A.R.C.S., F.Z.S.
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University,
Liverpool.
C. Tinting 6^ Co., Ltd.
Printers to the University Press of Liverpool
S3 Victoria Street
■83
FINAL REPORT OF THE LUANGWA
SLEEPING SICKNESS COMMISSION OF
THE BRITISH SOUTH AFRICA COMPANY
191 i - i 912
BY
ALLAN KINGHORN, WARRINGTON YORKE
AND
LLEWELLYN LLOYD,
ENTOMOLOGIST TO THE COMMISSION
(Received for publication 7 February , 1913)
Plates XV-XXVI
CONTENTS page
Introduction. 184
Section I. The Human Trypanosome—
(a) Distribution of the Disease. 185
(b) Clinical Features . 186
(c) Identity of Parasite with T. rbodesiense ... ... ... ... ... 187
(d) Transmission of the Trypanosome. 187
(e) Influence of Meteorological Conditions on Development of the Trypano¬
some in Glossina morsitans ... ... ... ... ... ... 204
(/) Reservoir of the Trypanosome ... ... ... ... ... ... 214
( g) Occurrence of the Trypanosome in Glossina morsitans in Nature ... 215
(b) Identity of 4 Game ’ and ‘ Fly ’ Strains with the ‘ Human * Strain of
T. rbodesiense ... ... ... ... ... ... ... ... 217
(s’) Summary . „.. 224
Section II. Trypanosomes or Game and Domestic Stock—
(1 a ) Methods . 227
(b) Examination of Game at Nawalia (Luangwa Valley) and Ngoa (Congo-
Zambesi Watershed). 228
(c) Examination of Domestic Stock . 235
(d) Summary . 237
Section III. Trypanosomes found in Wild Glossina morsitans —
(a) At Nawalia . 239
(b) At Ngoa . 241
(c) Summary . 243
Section IV. Description of Trypanosomes—
(a) Trypanosoma rbodesiense . 245
(b) Trypanosoma vivax ... ... ... ... ... ... ... ... 245
(c) Trypanosoma nanum . 248
(d) Trypanosoma pecorum ... ... ... ... ... ... ... 251
(e) Trypanosoma multiforme , sp. nov. 254
(/) Trypanosoma montgomeryi . 261
(g) Trypanosoma ignotum , sp. nov. ... ... ... ... ... ... 263
(b) Trypanosoma tragelapbi. sp. nov.* . 269
Section V. Development of Trypanosoma rbodesiense in Glossina morsitans — 273
Summary . 282
Section VI. Report of Entomologist—
(a) Glossina morsitans in the Laboratory ... ... ... ... ... 285
(b) Breeding Places of Glossina morsitans . 289
(c) Bloodsucking Insects collected at Nawalia and Ngoa . 293
Appendix
A. An Experiment to ascertain whether Tabanids transmit Trypanosomes
in Nature . 299
B. An Attempt to Transmit Trypanosoma rbodesiense by means of
Ornitbodoros moubata ... 301
184
INTRODUCTION
During the years 1909 and 1910, the diagnosis of several cases
of human trypanosomiasis amongst Europeans, who had never
been in contact with Glossina palpalis , drew attention to the
occurrence of the infection in portions of Rhodesia and Nyasaland
in which this particular insect was not known to exist. As the
result of prolonged and careful search, it was definitely proved that
Glossina palpalis did not occur in these areas, and accordingly, in
the beginning of 1911, this Commission was instituted by the
Chartered Company to ascertain the transmitting agent.
Nawalia, in the Luangwa Valley, was chosen as the site of the
laboratory (PI. XV, figs. 1-2). Work was commenced at the end of
June, 1911, and was continued until April, 1912. Our investiga¬
tions quickly placed it beyond doubt that Glossina morsitans was
the vector of the human trypanosome, and further revealed the fact
that a considerable percentage of game and of 1 wild * Glossina
morsitans were infected with the same parasite.
In April, 1912, the headquarters of the Commission were
removed to Ngoa, on the Congo-Zambesi watershed, in order that
experiments might be undertaken to ascertain what influence, if
any, was exerted by climatic conditions on the transmission of the
trypanosome. Work at Ngoa (PI. XVI, figs. 1-2) was continued
until the end of August, 1912, when the Commission left for
England. By this time it had been definitely determined that the
relatively low temperatures experienced during the cold season on
the plateau were inhibitory to the completion of the developmental
cycle of the’human trypanosome in Glossina morsitans.
Records of the greater portion of the work embodied in this
report have been published from time to time in the form of
separate papers. As, however, these preliminary communications
were necessarily somewhat incomplete, it was considered desirable
to collect and correlate all the results in the final report.
We desire to acknowledge our indebtedness to Dr. Aylmer May,
Principal Medical Officer, Northern Rhodesia; to Dr. A. F.
Wallace, M.O., N. Rhodesia; and to E. A. A. Jones, Esq.,
Assistant Magistrate, Mpika, for the great assistance they rendered
the Commission.
The report was completed at the Runcorn Research Laboratories
of the Liverpool School of Tropical Medicine.
i8 5
SECTION I
THE HUMAN TRYPANOSOME
BY
ALLAN KINGHORN
AND
WARRINGTON YORKE
<«) OCCURRENCE OF THE DISEASE
The distribution of human trypanosomiasis, due to infection
with T. rhodesiense , is comparatively wide. When the parasite was
first described,* the Luangwa Valley was the only region known
to be implicated, but it has since become apparent that the disease
is much more widely disseminated.
In the territory formerly known as North-Eastern Rhodesia,
cases have been diagnosed not only in the Luangwa Valley, but
also in the districts south of Fort Jameson and to the west of
Serenje. It may be mentioned further that this parasite was
isolated from wild game in the vicinity of Ngoa, and from a
native dog living in a village on the Nyasaland boundary. It is
possible, therefore, that sporadic cases of the disease may exist
in these localities also, but on this point no definite information
can be given.
In North-Western Rhodesia, it would appear that T. rhodesiense
exists in at least one locality, namely, between Broken Hill and
the Anglo-Belgian boundary. The details of a case, due to this
organism, and contracted in the district named, have recently been
described by Ellacombe.f
It is, of course, occasionally very difficult to ascertain the exact
locality in which the infection was acquired, more particularly in
native cases, but there are good grounds for believing that those
from the districts mentioned were autochthonous.
• Stephen! and Fantham, (1910). Roy. Soc. Proc., B, Vol. 83, p. 28. Annals of Trop.
Med. and Parasit., IV, p. 343.
t EUacombe, G. W. Sleeping Sickness Bulletin, 1912, Vol. IV, p. 185.
Beyond the confines of Northern Rhodesia, an instance of
infection by T . rhodesiense in Nyasaland has been recorded by
Stannus and Yorke,t an observation which has been confirmed more
recently by the Royal Society’s Commission. + The parasite has
been isolated from a case of sleeping sickness originating in
Portuguese East Africa, §, and it is accordingly probable that cases
reported from the neighbouring portions of German East Africa
are due to the same trypanosome. Finally, indigenous cases of
sleeping sickness have recently been diagnosed in Southern
Rhodesia, though we are not in a position to state the identity of
the organism.
It will be seen, therefore, that Trypanosoma rhodesiense is
widely spread over South Central Africa, and its distribution is in
close association with that of Glossina morsitans, which has been
shown to be the vector.
(A) CLINICAL FEATURES OF THE INFECTION
There is no essential difference between the clinical manifesta¬
tions of the disease caused by T. rhodesiense and that due to
T gantbiense> except possibly the greater virulence of the former.
In the earlier stages, enlargement of the lymphatic glands is
commonly seen, and more or less irregular fever, with the
accompanying symptoms of malaise, anorexia, headache, pains in
the limbs, &c., is the rule. During the attacks of fever trypano¬
somes can readily be demonstrated in the peripheral blood, but as
a rule are rather scanty. As the disease progresses oedemata of
various parts of the body, more particularly the face, are common
features, together with emaciation, loss of muscular power, ataxic
gait, tremors, difficult speech, loss of memory, and other signs of
nervous derangement. These symptoms gradually increase in
intensity until finally the patient sinks into a condition of coma,
and dies.
No exact data exist as to the duration of the disease in natives,
but it would appear to be short. Many of the cases complained of
t Stannua and Yorke. Proc. Rov. Soc., B 84, 1911, p. 156.
I Royal Society Commission. Rov. Soc. Proc., B 85, 1912, p. 423.
>: Sleeping Sickness Diary. Nyasaland Protectorate.
* 7
no subjective symptoms of the disease when diagnosed, and
presented very few objective signs, but in general they lived only
a few months. Occasionally, however, a patient is more resistant,
and one native definitely proved to be infected with T. rkodesiettse
is still alive and in a state of apparent good health a year later.
(c) IDENTITY OF TRYPANOSOME WITH T. RHODESIENSE
The essential characteristic of T . rhodesiense , Stephens and
Fantham, is the occurrence of ‘posterior nuclei/ i.e., amongst the
short forms of the parasite, organisms in which the macronucleus
is markedly displaced from the usual central to a decidedly
posterior position are commonly seen in certain laboratory animals,
more especially rats. This displacement may proceed to such a
degree that the macronucleus may actually be situated behind the
micronucleus at the aflagellar end of the trypanosome. As this
peculiarity has not been observed in T. gambiense , the two strains
of human trypanosomes can readily be distinguished. A further
difference is noticed in the pathogenicity of the parasites,
T. rhodesiense being much more virulent for all species of animals
than T. gambiense .
At Nawalia and Ngoa, rats were subinoculated from sixteen
cases of sleeping sickness, and in every instance the posterior
nuclei were observed in stained preparations. The pathogenicity
of these strains agreed closely with that of the Armstrong strain,*
from which T. rhodesiense was originally described.
It will be understood, therefore, that in all our transmission
experiments the strains of human trypanosomes utilised answered
in all respects to the description of T. rhodesiense .
(d). TRANSMISSION OF THE TRYPANOSOME
Experiments to transmit the human parasite by Glossina
morsitans were made at Nawalia in the Luangwa Valley, and at
Ngoa on the Congo-Zambesi watershed. Nawalia, the site of an
old station, is situated on the right bank of the Nyamadzi river,
Bevan. Journal of Comparative Path, and Therapeutics, 1910, p. 160.
Yorke. Annals of Tropical Medicine and Parasitology, 1910, p. 351.
a tributary of the Luangwa, at an altitude of about 2,100 feet above
the sea. Its position Is approximately 12 0 25' S. and 32 0 2' E.
Ngoa, ii° 40' S. and 31 0 30* E. lies some 30 miles North of the
station of Mpika, at a height of 4,400 feet above sea level.
The meteorological observations at Nawalia and Ngoa are
synopsised in Tables 1 and 2.
Certain general conditions which attach to all the experiments
may be mentioned.
The identity of the flies has been controlled both by direct
examination of the external characters and by the preparation of
the male genitalia, as recommended by Newstead,* so that we can
state with some degree of confidence that we have been dealing
only with Glossina morsitans, Westw.
All the experimental animals have been kept in fly-proof cages,
the fronts of which were protected by a double layer of wire gauze,
the inner composed of coarse, and the outer of mosquito meshing.
The two layers were separated by a space of one inch in order to
obviate the possibility of an animal being bitten while pressing its
body against the front of the cage.
The feeding of the flies, and the changing into fresh bottles
daily, was supervised personally, while the flies were kept in such a
manner that they had no opportunity of obtaining food from other
than the animals used in the actual experiments.
Experiments at Nawalia
A. With Laboratory-bred Glossina morsitans
Experiment 1
Commenced August 20th, 1911.
It is somewhat difficult to tabulate this experiment, owing tb the
fact that it was not started on a definite date with a definite number
of flies. Between August 20th and September 29th twenty-six flies
had hatched out, and each, as it did so, was given its first meal
on an animal showing numerous parasites in the peripheral blood,
so that on any given date the periods which had elapsed since the
infecting feeds of the flies varied considerably. In Table 3 the
main facts in connection with the flies are given.
• Newstead, R. Bull. Entomol. Research, Vol. II, Part I, May, 1911.
Annals Trof>. Med. G? Parasitol ., Vol. VII
PLATE XV
External Shade Temperatures Laboratory Temperatures Relative Days on
189
I9«
Tabu 3.—Giving date of infecting meal, date of death, and duration of life after infecting meal.
No. of fly
Date of infecting meal
Date on which fly died
Duration of life from
date of infection
A 1
20.8.11
22.9.11
33 days
A 2
21.8.11
20.1O.11
60 „
A 3
25.8.11
6.9.11
12 „
A 4
26.8.11
23.10.11
58 n
A 5
27.8.11
28.10.11
62 „
A 6
27.8.11
17.10.11
51 SI
A 7
28.8.11
11.10.11
44 1,
A 8
29.8.11
11.11.11
74 is
A 9
30.8.11
12.9.11
13 IS
A 10
31.8.11
20.10.11
5 ° is
A 11
31.8.11
12.9.11
1* n
A 12
3.9.11
27.9.11
24 s.
A 13
3 - 9 -n
1
5 - 9 -n
2 1,
A 14
5 - 9 -ii
K»
OO
vO
23 l,
A 15
6.9.11
25.IO.il
49 n
A 16
7 - 9 -ii
4 .II.II
58 n
A 17
8.9.11
3 . IO. I I
25 »
A 18
8.9.11
i4.IO.il
36 „
A 19
9 - 9 -ii
i9.IO.II
40 11
A 20
9 - 9-11
13.9.11
4 11
A 21
13.9.11
23.IO.II
40 n
A 22
16.9.11
14 . 11 .II
59 ,1
A 23
17.9.11
27.IO.I I
4 ° v
A 24
25.9.11
3-11*11
39 si
A 25
26.9.11
29.9.11
3 11
A 26
26.9.11
29.10.11
33 11
1
192
Twenty-four hours after each fly had fed on an infected animal,
it was afforded an opportunity of feeding on a clean monkey
(No. 41), after forty-eight hours on a second (No. 42), and from
the third day onwards on a third (No. 52). The schedule of
feedings is given in Table 4.
Table 4. —Showing transmission of human trypanosomes by laboratorv-bred Clossina moriitans.
Date
Animal
No. flics
Result
Remarks
fed
Aug. 21—Sept. 18...
Monkey
4 *
5
Negative
Flies fed 24 hours after infecting
feed.
„ 22 „ 28...
„
42
■ 4 *
Flies fed 48 hours after infecting
feed.
„ 23 „ 26...
52
23
Infection
Flics fed 72 hours and onwards
after infecting feed.
”
68
S
Negative
(<i) Infecting feed over 30 davs
before.
Sept. 27 anti 28 ...
69
6
(h) Infecting feed between 2oto 30
1
davs before.
1
..
70
S
Infection
(r) Infecting feed less than 20
davs before.
Sept. 29—Oct. 3 ...
72
16
All the flies fed.
Oct. 4 .
White rat
77
16
All the flics fed.
» 5 .
Monkey
68
*4
Negative
Infected fly did not feed.
>* 6—9 .
»»
58
16
Infection
All the flies fed.
» 9 .
61
»»
Infected flv, only, fed.
„ I! —12
n
68
*4
| Infected fly fed on 13th and 14th
as well.
„ 13—16
69
13
Negative
fflfected flv did not feed.
„ l6—19
83
12
Infection
Infected fly commenced feeding
on 16th, others on 17th.
1 Infected fly did not feed.
„ 20—28
n
69
10
Negative
„ 29—Nov. 11...
56
4
Infected fly did not feed.
• The remaining flics refused to feed.
From this table it will be seen that neither of the first two
monkeys on which the flies were allowed to feed became infected,
whereas No. 52 did so on the 27th September.
A reference to Table 3 will show that up to, and including the
26th September, twenty-three flies had fed on an infected
animal more than three days previously, and had, accordingly,
193
been fed on Monkey No. 52. The three flies, A 24, 25 and 26, had
never fed on this animal, and therefore had not to be considered
in the attempt to isolate the infected fly. Moreover, six of
the flies, A 1, 3, 9, u, 13 and 20, had died prior to the
26th September, and of these, three proved to be negative
throughout on examination. The other three, namely, A 3, 11 and
20, were found to show a heavy intestinal infection of trypano¬
somes. Fly A 3 died on September 6th, A 11 on September 12th,
and A 20 on September 13th, while the monkey did not become
infected until September 27th, much too long an incubation period
for one of these flies to have been the infecting one. We have
additional proof for the conclusion in that the abdominal contents
(gut + salivary glands) of flies A 3 and A 20, on inoculation into
monkeys, did not determine an infection.
On the 26th September there were, then, twenty flies with which
to deal, amongst which was at least one infective fly. As
stated above, three flies, A 24, 25 and 26, had never fed on
Monkey No. 52, so that the inquiry was limited to seventeen, and
this was further reduced to 15 by the death of flies A 12 and 14 on
September 27th and 28th—both flies negative on examination.
These were accordingly split up into three groups, based on
the length of time which had elapsed since the date of the
infecting feed, and each group was allowed to feed for two days
on a clean monkey, Nos. 68, 69 and 70.
Group ( a ) Infecting meal over thirty days previously.
,, ( b ) ,, ,, between twenty and thirty days
previously.
,, (c) ,, ,, less than twenty days previously.
Of the three monkeys, No. 70 was the only one to become infected,
and the transmitting fly was thus located in Group ( c ), consisting
of A 19, 21, 22 and 23.
While waiting to ascertain which of the three monkeys would
become infected, all the flies were fed from September 29th to
October 3rd on Monkey No. 72, and on October 4th on White Rat
No. 77. Both of these animals became infected in due course.
On October 5th all the flies, with the exception of A 19, were
re-fed on Monkey No. 68, and from the 6th to the 9th all were fed
on Monkey No. 58, except on October 9th when fly A 19 alone,
i 9 4
was fed on Monkey No. 61. Of these animals, No. 68 did not
become infected, while Nos. 58 and 61 did. The fly A 19 (c?) was
thus proved to be the infecting one.
No particular interest attaches to the further experiments. From
the table it will be seen that both those animals on which fly A 19
fed became infected (Nos. 68 and 83), whereas those on which it
did not feed remained quite healthy (Nos. 69 and 56).
When it had been definitely ascertained which was the infecting
fly, it was possible to determine fairly accurately the duration of
the cycle of the trypanosomes in the insect. Fly A 19 had its
infecting meal on September 9th, and Monkey No. 52, the first to
become infected, showed trypanosomes in the peripheral blood on
September 27th. On the 26th, therefore, the last day on which
the flies were fed on this animal, a period of eighteen days had
elapsed since A 19 fed on the infected animal. The average
incubation period of the local strain of human trypanosomes in
monkeys is four to five days, and subtracting this from the eighteen
days, it is evident that the fly must have become infective in thirteen
days.
This fly, A 19, lived for forty days from the date of the
infecting meal, and between the time of becoming capable of
transmitting the parasite and the date of death had fed on eight
animals, all of which became infected.
The other flies were fed continuously from the date of the
possible infecting meal to that of death, which occurred at varying
periods from two to seventy-four days, but none of them became
infective.
Experiment 2
Commenced November 14th, 1911, with sixteen laboratory-bred
Glossina morsitans.
In this experiment the flies were infected directly on a case of
human trypanosomiasis, each fly being allowed to feed on one
occasion only. Ten fed on the 14th of November, when the patient
showed three trypanosomes per field in the blood, and the remaining
six on the 15th, when there was, on an average, one trypanosome to
seven fields (Zeiss Oc. 4, Obj. D.D.). The subsequent meals were
as shown in Table 5.
'95
Table 5.—Showing transmission of human trypanosome by laboratory-bred Glossina morsitans .
Days after
infecting feed
Animal
No. of
flics fed
Result
Remarks
1 to 5
White rat
116
1 5
Negative
6 to 10
n
118
15
n
I! to 15
124
*5
Infection
16 to 20
»
116
*3
n
21 to 22
n
118
12
11
23 to 27
Monkey
137
6
Negative
) Flies divided into two groups to
1 separate the infective one.
23 to 27
n
00
5 *
Infection
28 to 42
”
*48
Varied
Negative
• One fly of this group (6) refused to feed, and died on the 26th day of the experiment.
Rat No. 124 became infected on December 4th, five days after
the flies had fed last, and as the incubation period of the trypano¬
some in these animals, on an average, is five days, it seems probable
that the fifteenth day was the one on which the infecting fly became
capable of transmitting the parasite.
On the 7th of December (23rd day after infecting meal)
the twelve flies then alive were divided into two groups in order to
effect an isolation of the infective one, and were fed on Monkey
Nos. 137 and 138, as indicated in the table. On December 12th,
the fly numbered B 23 (< 5 ) died, and on examination proved to be
heavily infected throughout the alimentary canal, and in the
salivary glands. No infection of the proboscis, however, was
observed.
The other flies were fed on a clean monkey until the 42nd day,
but without result.
Experiment 3.
Commenced December 29th, 1911, with twenty laboratory-bred
flies.
These flies were infected directly from a patient in whose
peripheral blood three trypanosomes per field (Zeiss Oc. 4,
196
Obj. D.D.) were seen. They were afterwards fed daily for sixty-
five days on a series of healthy monkeys, none of which became
infected. From the 67th—70th day of the experiment, the seven
flies then alive were fed on a guinea-pig heavily infected with the
human trypanosome, and were then fed for a further period of
thirty days on a clean monkey. This did not become infected.
Experiment 4
Commenced January 12th, 1912, with twenty-three laboratory-
bred flies.
These were fed for four days on a patient showing, on an
average, one trypanosome to three fields in the peripheral blood,
and afterwards on healthy monkeys, as indicated in Table 6.
Table 6. —Showing transmission of human trypanosome by laboratory-bred Glossina morsitans .
Day
Animal
No. of
flies fed
Result
Remarks
0 to 3
j Patient
23
1 —
4
-
—
—
Flies starved.
5 to 8
Monkey
237
22
| Negative
*) to 12
V
23S
22
13 to 16
V
240
1
i “
M
1
17 tO 20
-
254
20
Infection
!
21 tO 23
-
237
18
1
— ;
Died on 24th day.
24 !
240
' '7 1
Infection ;
25 to 29
260
1 9 ;
Negative
|
- Flics divided into two groups.
261
, 9
Infection
30 to 60 |
-
272
: 16—0
Negative
Infected fly did not feed.
On February 20th, the 29th day of the experiment, the
fly numbered D 18 died, and on dissection .was found to show a
massive intestinal infection of trypanosomes. Unfortunately, the
fly had been dead for some hours before it was examined, and it
was found impossible to dissect out the salivary glands. The
whole abdominal contents, therefore (gut and glands) were crushed
'97
up in normal saline solution and inoculated into a healthy monkey,
which became infected five days later. The disease ran a typical
course.
None of the other flies—dissected as they died—were found to
harbour trypanosomes in the proboscis, gut, or salivary glands.
In this instance the time which elapsed from the date of the
first infective meal until the date on which the fly became capable
of transmitting the trypanosome (allowing five days for the
incubation period in the monkey) was nineteen days.
B. By ‘ WILD ’ Glossina morsitans
Experiment 5
Commenced on November 14th, 1911, with ninety-eight ‘wild’
flies.
Prior to infecting these flies with the human trypanosome, they
were fed for three days, November I4th-i6th, on a healthy
monkey (No. 95), and for the next four days on a native fowl.
The monkey never became infected. From the 21st to the 24th of
November the insects then alive, fifty-seven in number, were fed on
an infected monkey showing twenty to thirty trypanosomes per
field in the peripheral blood, and were afterwards fed on healthy
animals, as in Table 7.
Table 7. —Resuit of feeding * wild 1 Glossina morsitans on dean monkeys, after a preliminary
meal on an animal infected with the human trypanosome.
Days after first
infecting feed
Animal
No. of
Hies fed
Result
Remarks
4 to 6
Monkey
,2 5
48
—
Monkey died on 7th day.
7 to 9
7?
I2 7
4*
Negative
10 to 13
«
130
34
Infection
14 to 16
1?
i«9
3*
77
17 to 18
77
140
7
Negative
Monkey died on 19th day.
17 to 23
*7
1 4 1
10
Infection
Flics divided into two groups to
isolate infected one.
20 tO Zj
57
1 44
4
V
198
This experiment was finished after the flies had fed on the
25th day, the flies being then killed and embedded.
The duration of the cycle of the parasites in the flies, in this
instance, would appear to be slightly over eleven days. The first
infecting meal was taken on November 21, and Monkey No. 130
showed parasites in the peripheral blood on December 7, a difference
of sixteen days. As stated already, the incubation period of this
trypanosome in monkeys is about five days, and by subtracting this
from the sixteen days, we obtain eleven for the duration of the
development cycle.
Experiment 6.
Commenced January 12, 1912, with forty-two freshly-caught
flies.
After being fed for one day cm a monkey infected with the
human trypanosome, and showing numerous parasites in the
peripheral blood, the flies were fed on a clean monkey for nine days.
They were then starved for one day, and subsequently allowed to
feed on clean monkeys and rats from the 1 ith to the 33rd day.
None of these animals became infected. The flies were dissected
as they died, and while trypanosomes were found in the gut and
proboscis of several, in no instance was an infection of the salivary
glands observed.
Experiment 7
Commenced January 12, 1912, with forty-two freshly-caught
flies.
The details of this experiment are exactly similar to those of
Experiment 6, with the exception that from the 1st to the 9th day
the flies were fed on a native fowl instead of on a monkey. They
were starved on the 10th day, as before, and afterwards fed on
clean monkeys and rats from the nth to the 38th day. None of
these animals became infected. Trypanosomes were found in the
proboscis and gut of several of the flies when dissected, but in no
case were the salivary glands implicated.
Experiment 8
Commenced February 14th, 1912, with 104 freshly-caught flies.
On the 13th of February, the flics were fed on a healthy monkey
199
which did not become infected, thus excluding the possibility that
they were already infected with the trypanosome. On the four
succeeding days they were fed on a guinea-pig infected with the
human trypanosome, and showing numerous parasites in the
peripheral blood, and afterwards on clean monkeys as indicated
in Table 8.
Table 8. —Showing transmission of the human trypanosome by freshly-caught Glossina morsitans.
Day
Animal
No. of
flies fed
Result
Remarks
4
5 to io
Monkey
269
98
i
Negative
Flies starved.
11
12
99
269
64
99
Flies starved.
IJ to 27
99
280
41
99
Died on 28th day. Flies divided
13 to 29
99
28l
47
Infection
into two groups, A and B.
28 to 29
99
286
33
Negative
Group A, only, fed.
30
99
269
17
Infection
Group B, only, fed.
30 to 38
99
300
10
Negative
| A 1
30 to 40
99
301
IO
99
I Flies of group A divided
f A 2 into 3 sub-groups, A 1,
30 to 40
99
302
12
99
>
u>
>
>
3 * 33
99
303
12
99
B 1
31 to 37
99
3°4
11
Infection
Flies in group B divided
■ B 2 into three sub-groups,
31 to 34
99
3 ° 5
12
Negative
B 1, B 2, and B 3.
B 3
34 to 39
99
310
I I
99
Sub-group B 1 fed.
35 to 52
99
3 i 5
12
99
Sub-group B 3 fed.
39 to 51
99
300
*5
99
Sub-group A 1 and B 2 fed.
41 to 52
99
301
28
1
99
Sub-group A 2, A 3, and B 1 fed.
The insects were dissected as they died, but only in one, the
infective fly, was an infection of the salivary glands observed,
though in a considerable number an infection of the proboscis and
gut was found.
200
The duration of the development cycle of the trypanosomes in
the fly would appear to be twenty-five days in this experiment.
The flies were fed for the first time on the infected guinea-pig
on February 14th, and the first monkey became infected on
March 15th, thirty days later. The average incubation period of
the disease in monkeys is five days, so that the cycle took twenty-five
days to complete.
Discussion of Results
In these transmission experiments, there are at least three sources
of error which must be considered, (1) accidental infection of the
experimental animals by other than the experimental flies,
(2) hereditary transmission of trypanosomes from infected female
flies to their progeny, and (3) natural infection in the experimental
animals.
(1) With regard to the first of these, the conditions under which
the experimental flies and animals were kept have been mentioned
already, and it seems more than improbable that accidental infection
would account for the unfailing regularity with which the animals
became infected after the infective flies had fed on them. More¬
over, in all our experiments, 644, such an occurrence as the
unexpected infection of an animal has not been observed.
(2) The number of bred flies which we have been able to obtain
has been too small to permit us to examine many of them prior to
use in the experiments, but such as were, have been found uniformly
free of infection. Stuhlmann,* Kleine.t and Bruce* with his
colleagues have examined large numbers of bred flies belonging to
the species Glossina brevipalpis, Glossina morsitans, and Glossina
palpalis , and are unanimous in the opinion that hereditary trans¬
mission of trypanosomes does not occur amongst the tsetse flies.
(3) With reference to the third point, we have used in the course
some 256 monkeys, and have never seen a naturally-occurring
trypanosome infection in any of them. Plasmodium kochi and
microfilaria have been observed, but beyond these, nothing.
• Stuhlmann. Arbeit aus d. Kaiser. Gesundheitsamte, Band XXVI, Heft 3, p. 374.
f Kleine. Deutsche med. Wochenschrift, No. 45, 1909.
j Bruce, Hamerton, Bateman and Mackie. Reports of S.S. Commission of the Royal Society,
No. 11, pp. 122-125.
201
Experiment 2 was specially devised to obviate the possibility of
error through the use of local monkeys (Cercopitkecus pygerytkrus).
The flies, as they were obtained, were fed on healthy, imported
rabbits which showed no signs of infection throughout; they were
infected directly from the human host; and they were then fed on
white rats. With the exception of Experiment 1, all the bred flies
used in the transmission work were infected directly on the human
host.
The trypanosomes transmitted by these flies were identical with
the human one, both morphologically and in their animal reactions.
There are certain points in connection with the experiments
which appear to be worthy of emphasis. The number of bred flies
which has been used in each is strikingly small, very much more so
than in any other similar work, of which the records are available.
In the four experiments a total of eighty-five was employed, and of
these three only became infective. A percentage of
The time occupied by the trypanosomes in completing their
cycle in the flies is also strikingly short, approximately two weeks
(thirteen, fifteen, eleven, nineteen and twenty-five days).
It may be pointed out, however, that all our estimations of the
latent periods of the trypanosomes in the flies represent the probable
durations only. Although the average incubation period in
monkeys is five days, this has been found to vary from three to
eight days, and it is possible, therefore, that the cycle may have
been slightly shorter, or longer, in any one instance.
Moreover, a further source of error is introduced in those
experiments in which the flies were fed on an infected animal for
more than a single day. It has yet to be determined whether only
a definite percentage of flies are inherently capable of transmitting
the disease, or whether any fly will do so provided that it has an
opportunity of feeding on an infected animal at some particular
time during its existence. If the latter alternative be correct, the
peculiar factors governing their infectability have still to be
ascertained. Assuming the first view to be correct, then the latent
period of the trypanosomes in the flies must date from the first
occasion on which the insects were fed on the infected animal, while,
if the second be correct, the latent period may date from any of the
meals on the infected animal.
202
So far as our results go, we have seen no indication of late
infection in any of our flies, although some of them have lived as
long as seventy-four days after the potentially infecting meal.*
All our results go to show that mechanical transmission of the
trypanosomes does not occur, that is, if a period of twenty-four
hours has elapsed since the infecting meal. We have not made any
experiments to ascertain whether infection could be accomplished
by interrupted feeding. This has been proved with various insects,
but practically, would account for very few, if any, cases of the
disease.
The infective flies have been found to retain the power of
transmitting the parasites during life, and do not require to feed
more than a single time on an animal in order to infect it, neither
do they require, prior to becoming infected, to feed more than once
on an animal suffering from trypanosomiasis.
With regard to Experiment No. 5, although only two flies were
definitely proved to transmit the trypanosome, infection of the
salivary glands was found, on dissection, in four others. As will
be seen in Section V, this is strong presumptive evidence that these
four were also infective.
Experiments at Ngoa
A. With Laboratory-bred Glossina morsitans
Experiment 9
Commenced on June 23rd, 1912, with nineteen laboratory-bred
flies.
They were fed for five days on a heavily-infected guinea-pig,
and afterwards for fifty-five days on a healthy monkey which did
not become infected.
B. By 1 WILD ’ Glossina morsitans
Experiment 10
Commenced on May 18th, 1912, with 116 ‘wild’ Glossina
morsitans , which had previously been shown to be non-infective by
being fed on a healthy monkey. The flies were fed for four days
on a heavily-infected guinea-pig and afterwards for sixty-seven
days on three healthy monkeys, none of which became infected.
# See, however, page 209.
Fig. 2. Laboratory at Ngoa, Congo-Zambesi Watershed
To face p. 202
C. Tinting Co., Ltd., Imp.
203
Experiment u
Commenced on June 13th, 1912, with ninety ‘wild 1 flies
previously proved not to be transmitting the human trypanosome.
They were allowed to feed for three days on a heavily-infected
guinea-pig, and were afterwards allowed to feed on a healthy
monkey until the 40th day of the experiment. This monkey did
not become infected.
On the 41st day of the experiment the flies then alive, forty-two
in number, were placed in an incubator, kept at a temperature of
85° F. Three flies were found to be infective eight days later.
Experiment 12
Commenced on June 14th, 1912, with 119 ‘wild' Glossina
morsitans , proved to be non-infective. The flies were fed from
the 1st to the 3rd day of the experiment on a heavily-infected
guinea-pig, and afterwards to the 60th day on a healthy monkey,
which did not become infected.
On the 61st day the thirty-eight remaining flies were placed in
the incubator kept at 83° F., and were fed from the 6ist-75th
day on a clean monkey. The monkey died on the 76th day without
becoming infected. However, on dissecting the flies, one was found
to harbour trypanosomes in the gut and salivary glands, and the
contents of these structures inoculated into healthy monkeys caused
the animals to become infected with T . rhodesiense.
Experiment 13
Commenced on July nth, 1912, with 176 ‘wild* Glossina
morsitans , previously shown not to harbour the human trypanosome.
These flies were fed for three days on a heavily-infected guinea-pig,
and afterwards from the 5th—51st day on healthy monkeys, none
of which became infected.
Experiment 14.
Commenced on July 24th, 1912, with 160 ‘wild* Glossina
morsitans , previously proved to be non-infective. From the
1st—4th day they were fed on a heavily-infected guinea-pig, and
from the 5th—36th on a healthy monkey, which did not become
infected.
(e) INFLUENCE OF METEOROLOGICAL CONDITIONS ON THE
DEVELOPMENT OF THE TRYPANOSOME IN GLOSSINA MORSITANS
At Nawalia, eight transmission experiments were made, four
with laboratory-bred, and four with 1 wild * Glossina morsitans. At
Ngoa, six experiments were carried out, five with ‘ wild * and one
with bred flies. It is unfortunate that the bred flies in the latter
series were not more numerous, but owing to the low temperature
the majority of the flies did not emerge from the puparia, and many
of those which did were malformed, and quickly died. In all the
experiments in which 1 wild * flies were used, however, the
possibility that they were already infected with the trypanosome
was excluded by first feeding them on healthy monkeys.
Synopses of the two series of experiments are given in
Tables 9 and 10.
It will be seen from these tables that whereas, in the Luangwa
Valley, Trypanosoma rhodesiense was successfully transmitted by
Glossina morsitans , all efforts in this direction on the Congo-
Zambesi watershed have been in vain. Of 330 flies used in the
valley experiments, six, and probably ten, became infective. The
larger figure is based on the number of salivary gland infections
found in the flies. Our experience indicates that the implication
of these structures is intimately connected with the ability of
Glossina morsitans to transmit T rypanosoma rhodesiense ,
and that until they are invaded by the organisms the
flies are non-infective. Salivary gland infections have been found
in all the flies which were capable of transmitting the parasite. In
Experiment No. 5 (Table 7) six flies were found to harbour
trypanosomes in the glands, but of these only two were actually
proved to transmit Trypanosoma rhodesiense . As in all other
instances, it was shown conclusively that those flies in which
trypanosomes were found in the salivary glands were infective,
it may be concluded that the remaining four flies in this experiment
were also capable of transmitting the parasite. Invasion of the
salivary glands has not been observed except in those flies which
were known to transmit Trypanosoma rhodesiense .
In the six plateau experiments, 680 Glossina morsitans were
employed without a single fly becoming infective.
“ im carried out at Nawnlia, L u .ngw» Valley, Southern Rhodesia.
205
Relative
humidity*
%
Ooooo N
Absolute
minimum
during
developmental
cycle
u-» y-k *j-> op 00 00 00 O
« N N N N N N N
Absolute
maximum
during
developmental
cycle
O N N N O
6 N in ir vO
^C'O'OOOOOOOOOO
Mean
temperature
during
developmental
cycle
-I w^oonl 00 O' en
M M 00 ob OO is f'v
N OO OO N N N N N
Duration of
developmental
cycle
days
ro u-> — | | | Os ^
Result
Infection
»
Negative
n
>>
Infection
»»
Variety of
flies used
Bred
Wild
Bred
Wild
«
Bred
WUd
No. of
flies used
VOVOrs.ONNf^l'if-
Season
m
k 1
v rj •*< ►% X
§ 0 q c s = p
0 li h
0 B
Date on
which
started
ooMMriMMMrj
N
No.
N N CO
*o
O
C
<£
"T3
0
3
JZ
-T3
s
-C
4 -»
a
206
The explanation of these apparently contradictory results is at
first sight not very obvious, more particularly in view of the fact
that even on the plateau ‘wild* Glossina morsitans capable of
infecting healthy monkeys with Trypanosoma rhodesiense were
occasionally encountered. If the climatic conditions under which
the valley experiments were carried out be compared with those
obtaining during the plateau experiments, it will be seen at once
that the most striking difference is one of temperature. As a rule,
the temperature during the former series of experiments was roughly
from 15-20 0 F. higher than during the latter series.
With a view to ascertaining the influence, if any, exerted by
temperature on the developmental cycle of Trypanosoma rhodesiense
in the tsetse fly, a further series of experiments were performed on
the plateau, in which, by means of an incubator, the flies were kept
at a temperature approximating to that of the valley at the most
favourable season.
In the first two experiments 1 wild * flies were used. No water
was placed in the incubator, and the warm dry air was found to
have a very deleterious effect on the insects. Within the
first seven days, twenty-five of the sixty-one flies with which
Experiment No. i was commenced, and fifty-three of the seventy-
two in Experiment No. 2 had died.
Notwithstanding the small number alive at the end of the
second week, two infective flies were obtained in the first experiment
and one in the second. In the third experiment laboratory-bred flies
were employed instead of ‘ wild ’ ones. Attention was drawn to the
fact that the low temperatures obtaining at Ngoa in the cold season
were very unfavourable to the pupation of Glossina morsitans; in
fact, so slow was the process that in spite of the large number of
pupae at our disposal, we were unable to procure sufficient flies for
experimental purposes. The difficulty was all the greater as many
of those which did emerge were malformed, and quickly died. In
order to obtain a sufficient number of bred flies for this experiment
we resorted to the expedient of placing the pupae, some of which
had been deposited over two months previously, in the incubator
(85° F.). Within three or four days a large number of flies were
procured.
The experiment was commenced on August 8th with thirty bred
20 7
flies, to which were added twelve on the 9th, eleven on the 10th, and
three on the nth. These groups were fed for four, three, two and
one days, respectively, on a heavily-infected guinea-pig and after¬
wards on a healthy monkey. Parasites were found in the blood of
the animal on which the flies of the fourth group were fed on the
18th after the insects fed on the guinea-pig, so that, allowing five
days for the incubation of the disease in the monkey, the fly was
infective on the 13th day. The monkey on which the flies in
Group 1 were fed became infected on the 26th day of the experi¬
ment, so that the duration of the developmental cycle of the parasite
in the fly would be twenty-one days. As we cannot be certain that
the infective fly fed on the infected guinea-pig on each of
the four days, the latent period in the insect may be anything
from seventeen to twenty-one days. The animals on which
Groups 2 and 3 were fed did not become infected.
It will be seen from Table 11 that the total number of flies used
in the incubator experiments was 189—133 ‘wild* and 56
' bred.’ Of the ‘ wild ’ flies, three became infective, and of the
‘bred’ two, i.e., 26% of the total number used. This figure is,
however, hardly a fair estimate, as of the 133 ‘wild’ flies only
seventy-eight were alive at the end of the first seven days of the
experiment. This heavy mortality was probably due to the sudden
change from the cold external air to the warm, dry atmosphere of
the incubator. As a general rule, in our transmission experiments
it was found that about 10 % of the flies died during the first week.
This was approximately the case in the incubator experiment in
which bred flies were used, as, owing to the fact that the insects
were hatched out in the incubator they were not subjected to any
sudden change of atmospheric conditions. We consider, therefore,
that had the mortality of the flies in these two experiments been the
customary 10% instead of over 40%, the proportion of infective
flies would be three of ninety instead of three of 133, or 3 3%
instead of 2 2%. This figure, 3 3, approximates closely to that
obtained in the incubator experiment in which bred flies were used,
namely, 3*5» <*nd «dso to that obtained in the valley experi¬
ments, 35.
The results of these three series of experiments, viz., those
carried out at Nawalia at laboratory temperatures 75°-84° F.,
198
This experiment was finished after the flies had fed on the
25th day, the flies being then killed and embedded.
The duration of the cycle of the parasites in the flies, in this
instance, would appear to be slightly over eleven days. The first
infecting meal was taken on November 21, and Monkey No. 130
showed parasites in the peripheral blood on December 7, a difference
of sixteen days. As stated already, the incubation period of this
trypanosome in monkeys is about five days, and by subtracting this
from the sixteen days, we obtain eleven for the duration of the
development cycle.
Experiment 6.
Commenced January 12, 1912, with forty-two freshly-caught
flies.
After being fed for one day cm a monkey infected with the
human trypanosome, and showing numerous parasites in the
peripheral blood, the flies were fed on a clean monkey for nine days.
They were then starved for one day, and subsequently allowed to
feed on clean monkeys and rats from the nth to the 33rd day.
None of these animals became infected. The flies were dissected
as they died, and while trypanosomes were found in the gut and
proboscis of several, in no instance was an infection of the salivary
glands observed.
Experiment 7
Commenced January 12, 1912, with forty-two freshly-caught
flics.
The details of this experiment are exactly similar to those of
Experiment 6, with the exception that from the 1st to the 9th day
the flies were fed on a native fowl instead of cm a monkey. They
were starved on the 10th day, as before, and afterwards fed on
clean monkeys and rats from the nth to the 38th day. None of
these animals became infected. Trypanosomes were found in the
proboscis and gut of several of the flies when dissected, but in no
case were the salivary glands implicated.
Experiment 8
Commenced February 14th, 1912, with 104 freshly-caught flies.
On the 13th of February, the flies were fed on a healthy monkey
209
those made in the laboratory at Ngoa at from 59°-65° F., and the
incubator experiments at Ngoa 8o‘6°-82‘6° F., show in a most
conclusive manner that comparatively high temperatures, 75°-85° F.,
are necessary for the completion of the developmental cycle of
Trypanosoma rhodesiense in Glossina morsitans.
In addition to temperature, there is another factor in the
climatic conditions which might possibly influence the develop¬
mental cycle of the trypanosome in Glossina morsitans. We refer
to the relative humidity of the atmosphere. At the most favourable
season of the year in the Luangwa Valley for transmission experi¬
ments, and also in the case of the first two carried out in the
incubator, the relative humidity was extremely low. In order to
decide the point, the relative humidity of the atmosphere in the
incubator in the bred fly experiment described above was kept at
from 70-72*5%. As the incubation period of the parasite in the
flies (twelve and seventeen to twenty-one days respectively) and also
the percentage of infective flies obtained (3*5 %) were approximately
the same as those in other experiments in which the relative
humidity was very low, we can only conclude that this factor does
not exert any appreciable influence on the developmental cycle of
T. rhodesiense in Glossina morsitans.
The following experiments were devised with a view to
ascertaining more definitely the influence of temperature on the
development of the parasite.
Experiment 15
Two batches of ‘ wild * Glossina morsitans (Batch A consisting
of 95 and Batch B of 119) in which the possibility of the
presence of an infective fly had previously been excluded by
feeding the insects on clean monkeys, were fed for three consecutive
days on a guinea-pig infected with T. rhodesiense. After being
starved for a day, each batch was fed on a healthy monkey until
the 40th day after the first feed on the infected animal. Neither
of the monkeys became infected. Batch A, in which there were
forty-two flies still alive, was placed in the incubator, whilst
Batch B, in which there* were now fifty-eight flies, was kept at
laboratory temperature. The sudden change from the laboratory
to the warm, dry air of the incubator proved very fatal to the flies
210
in Batch A, and on the 43rd day only six were alive. From the
41st to the 47th day the flies in this batch were fed on a monkey,
and from the 48th day on a rat. The rat became infected on the
53rd day, so that, allowing five days for the incubation of the
disease in the animal, Batch A contained an infective fly on the
48th day after the first feed on the infected guinea-pig, and eight
days after being placed in the incubator. As the monkey died on
the 47th day, we are unable to state whether the fly became infec¬
tive before the 48th day. The four flies still alive on the 53rd day
were fed on four clean rats, and three of these became infected.
The monkey on which Batch B was fed was still negative at the
end of sixty days, when there were thirty-eight flies alive.
Experiment 16
This is really a continuation of the former experiment. The
thirty-eight flies in Batch B were placed in the incubator on the
61st day after the first feed on the infected guinea-pig, and
were fed from the 6ist-75th day on a healthy monkey
(No. 443). Unfortunately the animal died on the 76th
day, so that we were unable to determine with certainty
whether any of the flies became infective. All the flies were
dissected as they died, and one was found to harbour trypanosomes
in the gut and salivary glands. Animals inoculated with the
contents of these structures became infected with T . rhodesiense.
As in our experience all flies in which salivary infection was
observed were capable of infecting animals with the human trypano¬
some, we may assume that had the monkey (No. 443) lived a few
days longer it would have been found to be infected.
In Experiment 15 the relative humidity of the air in the
incubator was very low (36 %), while in Experiment 16 the relative
humidity was comparatively high (72 %). In addition to confirming
the view that a relatively high temperature is essential to the
completion of the developmental cycle of T. rhodesiense in Glossina
morsitans , and that the relative humidity of the atmosphere is not
an important factor, these experiments afford more definite
information. It is apparent that the earlier stages of the develop¬
ment of the parasite in the fly can occur at comparatively low
temperatures (6o° F.), and that trypanosomes can persist in this
201
Experiment 2 was specially devised to obviate the possibility of
error through the use of local monkeys (Cercopithecus pygerytkrus ).
The flies, as they were obtained, were fed on healthy, imported
rabbits which showed no signs of infection throughout; they were
infected directly from the human host; and they were then fed on
white rats. With the exception of Experiment 1, all the bred flies
used in the transmission work were infected directly on the human
host.
The trypanosomes transmitted by these flies were identical with
the human one, both morphologically and in their animal reactions.
There are certain points in connection with the experiments
which appear to be worthy of emphasis. The number of bred flies
which has been used in each is strikingly small, very much more so
than in any other similar work, of which the records are available.
In the four experiments a total of eighty-five was employed, and of
these three only became infective. A percentage of 3*5.
The time occupied by the trypanosomes in completing their
cycle in the flies is also strikingly short, approximately two weeks
(thirteen, fifteen, eleven, nineteen and twenty-five days).
It may be pointed out, however, that all our estimations of the
latent periods of the trypanosomes in the flies represent the probable
durations only. Although the average incubation period in
monkeys is five days, this has been found to vary from three to
eight days, and it is possible, therefore, that the cycle may have
been slightly shorter, or longer, in any one instance.
Moreover, a further source of error is introduced in those
experiments in which the flies were fed on an infected animal for
more than a single day. It has yet to be determined whether only
a definite percentage of flies are inherently capable of transmitting
the disease, or whether any fly will do so provided that it has an
opportunity of feeding on an infected animal at some particular
time during its existence. If the latter alternative be correct, the
peculiar factors governing their infectability have still to be
ascertained. Assuming the first view to be correct, then the latent
period of the trypanosomes in the flies must date from the first
occasion on which the insects were fed on the infected animal, while,
if the second be correct, the latent period may date from any of the
meals on the infected animal.
212
stage for at least sixty days. It is obvious that the developmental
cycle of the parasite is not complete, since the flies are non-
infective, and inoculation of the gut contents into susceptible
animals is followed by negative results. For the completion of the
cycle it is necessary for the temperature to which the flies are
subjected to be raised to a considerable extent (75°~85° F.).
It is interesting to note that the flies in Batch A (Experiment 15)
became infective eight days, possibly less, after being placed in
the incubator. This is three days less than the shortest incubation
period observed in any of our successful transmission experiments,
a fact which supports the view that the developmental cycle of the
parasite in the fly had proceeded to a certain point at laboratory
temperature (6o° F.) before the insects were subjected to the higher
temperature (8o° F.) of the incubator.
The fact that an occasional infective ‘ wild * fly was encountered
on the plateau during a period (May, June and July) when attempts
to transmit in the laboratory were invariably unsuccessful requires
some explanation. A possible solution may be that the flies in
question were infected during the warmer season of the year and
had survived into the cold season.
If the results obtained by feeding freshly-caught flies on healthy
monkeys in the valley are compared with those from flies caught
on the plateau, a marked difference in the number of infections
resulting is apparent. In the Luangwa Valley, 3,202 flies were fed
in twenty-nine batches, and Trypanosoma rhodesiense was isolated
in six of the experiments, giving a ratio of 1 infective fly to 534,
whereas on the Congo-Zambesi watershed, 5,250 freshly-caught
Glossina morsitans were fed in groups on forty-one monkeys, with
four positive results—1 infective fly to 1,312. As tsetse flies and
game are about equally numerous at Nawalia and Ngoa, and as the
disease was presumably introduced into the two localities, which
are less than seventy miles apart, about the same time, it appears
to us that the only essential difference which can account for the
fact that the percentage of infective ‘wild' flies at Nawalia is two
and a half times as great as at Ngoa is the difference in the
climatic conditions. It will be seen from Tables 1 and 2 that the
temperatures experienced on the Congo-Zambesi watershed during
May, June and July, are very much lower than those at Nawalia
Annals Trop. Med. Parasitol., Vol. I'll
PLATE XVI
Fig. 2. Laboratory at Ngoa, Congo-Zambesi Watershed.
To face f. 202
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Feeding Glossina morsitans on experimental animal:
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eding Glossina morsitans on experimental animals.
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PLATE XVII
• T
i
203
Experiment n
Commenced on June 13th, 1912, with ninety ‘wild* flies
previously proved not to be transmitting the human trypanosome.
They were allowed to feed for three days on a heavily-infected
guinea-pig, and were afterwards allowed to feed on a healthy
monkey until the 40th day of the experiment. This monkey did
not become infected.
On the 41st day of the experiment the flies then alive, forty-two
in number, were placed in an incubator, kept at a temperature of
85° F. Three flies were found to be infective eight days later.
Experiment 12
Commenced on June 14th, 1912, with 119 'wild* Glossina
morsitans, proved to be non-in feet ive. The flies were fed from
the 1st to the 3rd day of the experiment on a heavily-infected
guinea-pig, and afterwards to the 60th day on a healthy monkey,
which did not become infected.
On the 61st day the thirty-eight remaining flies were placed in
the incubator kept at 83° F., and were fed from the 6ist-75th
day on a clean monkey. The monkey died on the 76th day without
becoming infected. However, on dissecting the flies, one was found
to harbour trypanosomes in the gut and salivary glands, and the
contents of these structures inoculated into healthy monkeys caused
the animals to become infected with T. rhodesiense.
Experiment 13
Commenced on July nth, 1912, with 176 ‘wild’ Glossina
morsitans , previously shown not to harbour the human trypanosome.
These flies were fed for three days on a heavily-infected guinea-pig,
and afterwards from the 5th—51st day on healthy monkeys, none
of which became infected.
Experiment 14.
Commenced on July 24th, 1912, with 160 ‘wild* Glossina
morsitans, previously proved to be non-infective. From the
1st—4th day they were fed on a heavily-infected guinea-pig, and
from the 5th—36th on a healthy monkey, which did not become
infected.
214
discrepancy may be explained on the assumption that the flies
became infective during the warm season and that a certain number
survived into the colder season of the year.
(/) THE RESERVOIR OF THE TRYPANOSOME
The possibility that game might act as a reservoir of infection
of sleeping sickness areas has been recognised almost since the
inception of work on the disease, but up to the present it would
appear that the trypanosomes have never been demonstrated in
such animals under natural conditions. In Uganda, Bruce,
Hamerton and Bateman* have proved that certain species of buck,
notably waterbuck, bushbuck, and reedbuck, can readily be infected
with Trypanosoma gambiense by allowing infected Glossina
palpalis to feed on them, and that healthy flies, in turn, may be
infected from game harbouring parasites in their blood. They
were unable, however, to examine a sufficiently large number of
head to ascertain whether a natural infection was present.
The importance of the question is obvious, and the results of our
investigations on the point afford a striking commentary on the
potential danger involved in the infection of the game.
The Luangwa Valley is particularly rich in a widely-varied
fauna, and owing to the fact that in the dry season the great bulk
of the game tends to collect in the vicinity of the few permanent
streams, it has been comparatively simple to shoot buck for the
purposes of experimentation. At Ngoa, on the Congo-Zambesi
watershed, game is plentiful at certain seasons of the year.
Trypanosomes indistinguishable from T. rhodesiense were
isolated from buck in both these localities, and, in addition, from
one native dog living in a village some fifty miles to the East of
Nawalia.
In the Luangwa Valley 127 head of game were examined, and
sub-inoculations into healthy monkeys and rats were made from 56.
In this manner the human parasite was recovered from the following
animals: —
4 Waterbuck,
1 Hartebeest,
2 Mpala,
1 Bushbuck,
I Wart hog.
•Bruce, Hamerton and Bateman. Proc. Roy. Soc., B, Vol. 83, 1911.
215
On the plateau 124 head were examined, and sub-inoculations
were made from sixty. The trypanosome was isolated from two
waterbuck only.
There is thus a marked difference in the percentage of game
infected with T. rhodesiense in the two districts, the parasite being
five times as frequent in the valley as in the plateau game.
As the game was shot without discrimination, the figures are
probably a fair indication of the proportion of the total game
infected in the two areas.
Tabli 14. —Percentage of game infected with T. rhodesiense .
Nawalia Ngoa
(Luangwa Valley) (Congo-Zambesi Watershed)
No. inoculations
made
No. infected
%
No. inoculations
made
No. infected
%
5 *
9
16
60
2
3*3
As stated above, the trypanosome was isolated from one native
dog only of thirty-five domestic animals examined—cattle, goats
and dogs.
Our experience indicates that big game is much the most
important reservoir of the infection. During our sojourn in
the country some 256 monkeys, 142 wild rats, and 15 wild mice
were examined, with negative results. In all, therefore, 698
animals were examined.
Apart from the bigger species of game, most of the smaller
wild animals are nocturnal in their habits, and it seems unlikely
that many of them would be exposed to infection.
(*) OCCURRENCE OP THE TRYPANOSOME IN CLOSSINA
MORSITANS IN NATURE
Series of experiments were carried out in the Luangwa Valley
and on the plateau to determine the species of trypanosome
transmitted, in nature, by Glossina tnorsilans.
A number of fly-boys were sent out from time to time to capture
and bring into the laboratory ‘ wild ’ tsetse flies, which were allowed
to feed on healthy monkeys. Full details of these experiments will
be found in a later section of this report.
At Nawalia (Luangwa Valley) the human trypanosome was
198
This experiment was finished after the flies had fed on the
25th day, the flies being then killed and embedded.
The duration of the cycle of the parasites in the flies, in this
instance, would appear to be slightly over eleven days. The first
infecting meal was taken on November 21, and Monkey No. 130
showed parasites in the peripheral blood on December 7, a difference
of sixteen days. As stated already, the incubation period of this
trypanosome in monkeys is about five days, and by subtracting this
from the sixteen days, we obtain eleven for the duration of the
development cycle.
Experiment 6.
Commenced January 12, 1912, with forty-two freshly-caught
flies.
After being fed for one day cm a monkey infected with the
human trypanosome, and showing numerous parasites in the
peripheral blood, the flies were fed on a clean monkey for nine days.
They were then starved for one day, and subsequently allowed to
feed on clean monkeys and rats from the 1 ith to the 33rd day.
None of these animals became infected. The flies were dissected
as they died, and while trypanosomes were found in the gut and
proboscis of several, in no instance was an infection of the salivary
glands observed.
Experiment 7
Commenced January 12, 1912, with forty-two freshly-caught
flics.
The details of this experiment are exactly similar to those of
Experiment 6, with the exception that from the 1st to the 9th day
the flies were fed on a native fowl instead of cm a monkey. They
were starved on the 10th day, as before, and afterwards fed on
clean monkeys and rats from the nth to the 38th day. None of
these animals became infected. Trypanosomes were found in the
proboscis and gut of several of the flies when dissected, but in no
case were the salivary glands implicated.
Experiment 8
Commenced February 14th, 1912, with 104 freshly-caught flies.
On the 13th of February, the flies were fed on a healthy monkey
217
(A) IDENTITY OF 'GAME* AND ‘FLY’ STRAINS^WITH THE
‘ HUMAN ’ STRAIN OF T. RHODESIENSE
The conclusion that one of the trypanosomes isolated from
game and from naturally-infected tsetse flies is identical with
T. rhodesiertse has been based on a careful study of the morphology
and pathogenicity of the three strains in question.
I. Morphology (PI. XVIII)
In fresh preparations, all three strains show the same mixture of
short, slowly-moving, and long, active forms, the relative numbers
of which vary in the peripheral blood of any animal from day
to day.
In stained preparations, it is sufficient to say that it is impossible
to distinguish any one of the three strains from the others. Short
forms in which the macronucleus lies actually posterior to the
blepharoplast have been observed in each of the three strains.
The measurements of the three strains also show an extremely
close agreement. Eleven hundred individuals of each have been
measured, and the results are given in Tables 16, 17 and 18. The
total number of parasites drawn from each variety of laboratory
animal is the same in the case of each strain, and only twenty-five
have been measured from any one preparation, as it has been found
that the average length varies within wide limits, from day to day,
in any given animal.
We are of the opinion that the measurement of twenty-five
individuals from one preparation gives a fair estimate of the
average length of the trypanosome present in the particular blood
film, and that the error is less if twenty-five parasites are measured
on each of forty-four different days than if, for example, 100 are
measured on each of eleven different days.
In measuring the parasites the following technique was
adopted:—Thin blood smears, dried in the air, were fixed in
absolute alcohol and stained with Giemsa’s solution. The trypano¬
somes were then outlined at a magnification of 2,000 diameters with
the aid of an Abbe camera lucida, and the length along the middle
line of the body measured by the tangent method described by
Stephens and Fantham.*
• Roy. Soc. Proc., B, Vol. 85, p. 223 (1912).
200
The duration of the development cycle of the trypanosomes in
the fly would appear to be twenty-five days in this experiment.
The flies were fed for the first time on the infected guinea-pig
on February 14th, and the first monkey became infected on
March 15th, thirty days later. The average incubation period of
the disease in monkeys is five days, so that the cycle took twenty-five
days to complete.
Discussion of Results
In these transmission experiments, there are at least three sources
of error which must be considered, (1) accidental infection of the
experimental animals by other than the experimental flies,
(2) hereditary transmission of trypanosomes from infected female
flies to their progeny, and (3) natural infection in the experimental
animals.
(1) With regard to the first of these, the conditions under which
the experimental flies and animals were kept have been mentioned
already, and it seems more than improbable that accidental infection
would account for the unfailing regularity with which the animals
became infected after the infective flies had fed on them. More¬
over, in all our experiments, 644, such an occurrence as the
unexpected infection of an animal has not been observed.
(2) The number of bred flies which we have been able to obtain
has been too small to permit us to examine many of them prior to
use in the experiments, but such as were, have been found uniformly
free of infection. Stuhlmann,* Kleine,t and Bruce* with his
colleagues have examined large numbers of bred flies belonging to
the species Glossina brevipalpis, Glossina morsitans , and Glossina
palpalis y and are unanimous in the opinion that hereditary trans¬
mission of trypanosomes does not occur amongst the tsetse flies.
(3) With reference to the third point, we have used in the course
some 256 monkeys, and have never seen a naturally-occurring
trypanosome infection in any of them. Plasmodium kochi and
microfilaria have been observed, but beyond these, nothing.
• Stuhlmann. Arbeit aus d. Kaiser. Gesundheitsamte, Band XXVI, Heft 3, p. 374.
f Kleine. Deutsche med. Wochenschrift, No. 45, 1909.
} Bruce, Hamerton, Bateman and Mackie. Reports of S.S. Commission of the Royal Society,
No. 11, pp. 122-125.
201
Experiment 2 was specially devised to obviate the possibility of
error through the use of local monkeys (Cere 0 pit hecus pygerytkrus ).
The flies, as they were obtained, were fed on healthy, imported
rabbits which showed no signs of infection throughout; they were
infected directly from the human host; and they were then fed on
white rats. With the exception of Experiment 1, all the bred flies
used in the transmission work were infected directly on the human
host.
The trypanosomes transmitted by these flies were identical with
the human one, both morphologically and in their animal reactions.
There are certain points in connection with the experiments
which appear to be worthy of emphasis. The number of bred flies
which has been used in each is strikingly small, very much more so
than in any other similar work, of which the records are available.
In the four experiments a total of eighty-five was employed, and of
these three only became infective. A percentage of 3*5.
The time occupied by the trypanosomes in completing their
cycle in the flies is also strikingly short, approximately two weeks
(thirteen, fifteen, eleven, nineteen and twenty-five days).
It may be pointed out, however, that all our estimations of the
latent periods of the trypanosomes in the flies represent the probable
durations only. Although the average incubation period in
monkeys is five days, this has been found to vary from three to
eight days, and it is possible, therefore, that the cycle may have
been slightly shorter, or longer, in any one instance.
Moreover, a further source of error is introduced in those
experiments in which the flies were fed on an infected animal for
more than a single day. It has yet to be determined whether only
a definite percentage of flies are inherently capable of transmitting
the disease, or whether any fly will do so provided that it has an
opportunity of feeding on an infected animal at some particular
time during its existence. If the latter alternative be correct, the
peculiar factors governing their infectability have still to be
ascertained. Assuming the first view to be correct, then the latent
period of the trypanosomes in the flies must date from the first
occasion on which the insects were fed on the infected animal, while,
if the second be correct, the latent period may date from any of the
meals on the infected animal.
202
So far as our results go, we have seen no indication of late
infection in any of our flies, although some of them have lived as
long as seventy-four days after the potentially infecting meal.*
All our results go to show that mechanical transmission of the
trypanosomes does not occur, that is, if a period of twenty-four
hours has elapsed since the infecting meal. We have not made any
experiments to ascertain whether infection could be accomplished
by interrupted feeding. This has been proved with various insects,
but practically, would account for very few, if any, cases of the
disease.
The infective flies have been found to retain the power of
transmitting the parasites during life, and do not require to feed
more than a single time on an animal in order to infect it, neither
do they require, prior to becoming infected, to feed more than once
on an animal suffering from trypanosomiasis.
With regard to Experiment No. 5, although only two flies were
definitely proved to transmit the trypanosome, infection of the
salivary glands was found, on dissection, in four others. As will
be seen in Section V, this is strong presumptive evidence that these
four were also infective.
Experiments at Ngoa
A. With Laboratory-bred Glossina morsitans
Experiment 9
Commenced on June 23rd, 1912, with nineteen laboratory-bred
flies.
They were fed for five days on a heavily-infected guinea-pig,
and afterwards for fifty-five days on a healthy monkey which did
not become infected.
B. By * WILD * Glossina morsitans
Experiment 10
Commenced on May 18th, 1912, with 116 ‘wild* Glossina
morsitans , which had previously been shown to be non-infective by
being fed on a healthy monkey. The flies were fed for four days
on a heavily-infected guinea-pig and afterwards for sixty-seven
days on three healthy monkeys, none of which became infected.
# See, however, page 209.
221
Table 19. —Comparison of the measurements of the * human/ 4 game,* and 4 fly * strains.
Strain
Length in microns
Average
Maximum
Minimum
‘ Human *.
21-25
39 '°
! 3’ 2 5
4 Game * .
21-38
35-5
m -75
‘Fly’ .
21-67
36-25
13*0
The similarity in the measurements is, perhaps, best appreciated
by a glance at the curves obtained by plotting out the distribution
of the various lengths of the parasites, expressed in percentages of
the total numbers measured (see Chart I).
A comparison of the percentages of 1 short and stumpy/ 1 inter¬
mediate * and ' long ’ forms is also of interest.
Table 20. —Comparison of percentage* of 1 short and stumpy/ 4 intermediate/ and 4 long * forms of
the 4 human/ 4 game/ and 4 fly * strains.
Strain
Short and stumpy
forms
II- 2 I/A
Intermediate forms
22-24/A
Long forms
25 - 39 M
4 Human *.
6478
15-98
19 -I 4
4 Game * .
62-87
1 5*34
2I-56
4 Fly ’ .
58-68
18*81
22‘4I
2. Pathogenicity
The pathogenicity of the three strains is synopsised in Table 21,
p. 223. A glance at this table will show how remarkably the three
strains agree in this respect.
Chast I. Companion of curvet of the 'human/ 'game,' and 'fly' strains by plotting out the distribution of the various
the parasites, expressed in percentages of the total numbers measured (1,100 of each strain).
At I CHONS
203
Experiment n
Commenced on June 13th, 1912, with ninety ‘wild 1 flies
previously proved not to be transmitting the human trypanosome.
They were allowed to feed for three days on a heavily-infected
guinea-pig, and were afterwards allowed to feed on a healthy
monkey until the 40th day of the experiment. This monkey did
not become infected.
On the 41st day of the experiment the flies then alive, forty-two
in number, were placed in an incubator, kept at a temperature of
85° F. Three flies were found to be infective eight days later.
Experiment 12
Commenced on June 14th, 1912, with 119 ‘wild' Glossina
morsitans , proved to be non-infective. The flies were fed from
the 1st to the 3rd day of the experiment on a heavily-infected
guinea-pig, and afterwards to the 60th day on a healthy monkey,
which did not become infected.
On the 61st day the thirty-eight remaining flies were placed in
the incubator kept at 83° F., and were fed from the 6ist-75th
day on a clean monkey. The monkey died on the 76th day without
becoming infected. However, on dissecting the flies, one was found
to harbour trypanosomes in the gut and salivary glands, and the
contents of these structures inoculated into healthy monkeys caused
the animals to become infected with T. rhodesiense .
Experiment 13
Commenced on July nth, 1912, with 176 ‘wild’ Glossina
morsitans , previously shown not to harbour the human trypanosome.
These flies were fed for three days on a heavily-infected guinea-pig,
and afterwards from the 5th—51st day on healthy monkeys, none
of which became infected.
Experiment 14.
Commenced on July 24th, 1912, with 160 ‘wild* Glossina
morsitans , previously proved to be non-infective. From the
1st—4th day they were fed on a heavily-infected guinea-pig, and
from the 5th—36th on a healthy monkey, which did not become
infected.
224
SUMMARY
1. The human trypanosome ( T . rhodesiense') is distributed
widely throughout South Central Africa.
2. There is no essential difference between the clinical manifes¬
tations of the disease in man caused by T. rhodesiense and that due
to T. gambiense 9 except possibly the greater virulence of the former.
3. T. rhodesiense is transmitted in Rhodesia by Glossitta
morsitans.
4. Approximately 3 5% of the flies may become permanently
infected and capable of transmitting the virus.
5. The period which elapses between the infecting feed of the
flies and the date on which they become infective varies from
eleven to twenty-five days in the Luangwa Valley.
6. Attempts carried out at laboratory temperature on the
Congo-Zambesi plateau during the cold season to transmit the
human trypanosome by means of Glossina morsitans were invariably
unsuccessful in spite of the fact that 680 flies were used in these
experiments.
7. The developmental cycle of T. rhodesiense in Glossina
morsitans is to a marked degree influenced by the temperature to
which the flies are subjected. High temperatures (75-85° F.)
favour the development of the parasite, whilst low temperatures
(6o°-70° F.) are unfavourable.
8. The first portion of the developmental cycle can proceed
at the lower temperatures, but for its completion the higher
temperatures are essential.
9. The parasites may persist in the fly at an incomplete stage
of their development for at least sixty days under unfavourable
climatic conditions.
10. These observations afford an adequate explanation of the
extremely long latent periods of trypanosomes in Glossina which
have occasionally been observed by various workers.
11. The relative humidity of the atmosphere has apparently no
influence on the development of the trypanosome in Glossina
morsitans ,
22 $
12. Mechanical transmission does not occur if a period of
twenty-four hours has elapsed since the infecting meal.
13. Glossina morsitans , in nature, has been found to transmit
the human trypanosome.
14. The chief reservoir of the human trypanosome is the
antelope.
15. The results of examination for the human trypanosome of
the blood of a large number of monkeys, wild rats and mice were
invariably negative.
Ii6
isolated in six of twenty-nine experiments, in which 3,202 tsetse
flies were used. Possibly it was present in a seventh, but as the
monkey died on the day after becoming infected, no definite state¬
ment can be made other than that the incubation period of the
disease was the same as that in known T. rhodesiense infections.
At Ngoa (Congo-Zambesi watershed) 5,250 freshly-caught
Glossina morsitans were fed in forty-two batches on healthy
monkeys, and the trypanosome was isolated on four occasions.
The ratio of infective to non-infective flies in the two localities,
assuming that only one was capable of transmitting the virus in
each instance, is, therefore,
At Nawalia, At Ngoa,
1 : 534. 1 : 1,312.
No definite comparison can be made between these figures, as
the experiments were not carried out under identical conditions.
Those at Nawalia were made during both the dry and wet seasons,
while those at Ngoa were carried out during the height of the
winter at a time when it was impossible to transmit the human
trypanosome in the laboratory. However, the difference is so
marked that it may safely be concluded that the plateau flies are
infective, in nature, to a much smaller extent, than those in the
Luangwa Valley.
In one of these experiments the actual infective fly was isolated.
Experiment 17
Commenced October 30th, 1911, with sixty freshly-caught flies,
to which were added twenty-two additional ones on the next day.
The flies were fed as indicated in Table 15.
On November 13th, the flies still alive, thirteen in number, were
killed and embedded. In the sections, numerous parasites were
found in the gut and salivary gland of only one of them.
Table 15.—Showing the transmission of the human trypanosome by naturally-infected Glossina
morsitans.
Date
Animal
No. of
flies fed
Result
Remarks
Oct. 30—Nov. 4 ...
Monkey
96
60-22
Infection
Nov. 6 .
105
29
Nov. 7—10
108
*9
Nov. 11—12
113
7 I
Negative ]
Flies divided into two
Nov. 11—12
"4
6
Infection V
groups to isolate the
infected fly.
217
( h) IDENTITY OF ‘GAME’ AND ‘FLY’ STRAINS^WITH THE
• HUMAN ’ STRAIN OF T. RHODESIENSE
The conclusion that one of the trypanosomes isolated from
game and from naturally-infected tsetse flies is identical with
T. rhodesiense has been based on a careful study of the morphology
and pathogenicity of the three strains in question.
i. Morphology (PI. XVIII)
In fresh preparations, all three strains show the same mixture of
short, slowly-moving, and long, active forms, the relative numbers
of which vary in the peripheral blood of any animal from day
to day.
In stained preparations, it is sufficient to say that it is impossible
to distinguish any one of the three strains from the others. Short
forms in which the macronucleus lies actually posterior to the
blepharoplast have been observed in each of the three strains.
The measurements of the three strains also show an extremely
close agreement. Eleven hundred individuals of each have been
measured, and the results are given in Tables 16, 17 and 18. The
total number of parasites drawn from each variety of laboratory
animal is the same in the case of each strain, and only twenty-five
have been measured from any one preparation, as it has been found
that the average length varies within wide limits, from day to day,
in any given animal.
We are of the opinion that the measurement of twenty-five
individuals from one preparation gives a fair estimate of the
average length of the trypanosome present in the particular blood
film, and that the error is less if twenty-five parasites are measured
on each of forty-four different days than if, for example, 100 are
measured on each of eleven different days.
In measuring the parasites the following technique was
adopted:—Thin blood smears, dried in the air, were fixed in
absolute alcohol and stained with Giemsa’s solution. The trypano¬
somes were then outlined at a magnification of 2,000 diameters with
the aid of an Abbe camera lucida, and the length along the middle
line of the body measured by the tangent method described by
Stephens and Fantham.*
• Roy. Soc. Proc., B, Vol. 85, p. 223 (1912).
T*able 16.—Giving details of measurement of i,ioo individuals of the * human * strain.
Animal
Day of
disease
Number
measured
Length in microns
f
Average
Maximum
Minimum
Monkey
5
6
25
21*03
27*75
*5*5
n
5
11
25
*9-5
2619
*3' 2 7
»>
6
8
25
*9*4*
28-0
*3*5
6
...
*5
25
26*3
3**5
J9-67
n
20
9
25
22*3
30-3
*7* *
n
20
*3
25
I 9 *97
26-25
13-25
n
*5
9
25
21*28
28*25
18-0
n
25
*3
25
*9'57
29-75
15-25
n
33
8
25
2 4 -2
28-75
16-75
?»
33
9
25
20-59
29*75
I5-29
»»
»7
11
25
22*8l
3**5
I8-25
11
87
21
25
22‘4I
29*25
.8-5
11
87
22
25
*9-95
2 7 -C
17-0
Dog
244
6
25
22-26
29 25
1875
ii
244
8
25
20-16
24*5
*7*5
ii
244
*3
25
2172
3*25
18-25
ii
244
*4
25
*97
22-5
*7-25
Rabbit
13
4
25
23*5
30-5
*4*5
ii
>3
22
25
18-11
*4-75
*5*5
ii
A
24
25
*9-52
39*5
*4*5
ii
86
*3
25
21-91
29-0
16-75
Guinea-pig
14
*4
25
21-09
30-25
*7*5
ii
H
20
25
22*03
3*75
16-o
ii
>4
22
25
22-21
33*25
*4*5
ii
*39
25
25
20*66
27-25
*575
ii
*39
36
25
18-2
26-5
140
ii
*39
52
25
; *8*4
28-0
*375
Rat
*5
22
25
20-03
25*5
*575
ii
*5
26
25
21-08
28-25
1675
ii
16
*5
2$
22-98
33*25
15-70
ii
*83
12
25
22-44
30*75
1875
ii
184
12
25
22*12
3**25
18*0
ii
184
*4
25
19-64
31*0
14-25
ii
*84
20
25
20-17
27*5
17*0
ii
184
28
25
20-59
30*0
1675
ii
208
10
25
19-32
23*25
1675
ii
208
20
25
19-69
24*5
*7-0
ii
212
6
25
23*33
31-0
, 8-75
ii
212
7
25
20-88
32*5
•55
ii
212
16
25
18-66
22*5
•375
Mouse
27
12
25
20*95
26-75
.8-5
28
6
25
*9*94
23-0
17-25
ii
9*
6
25
23-94
27*25
i8-5
ii
9*
10
25
28-65 j
33-0
21-5
IIOO
21-25
39-0
13-25
219
Table 17.—Giving details of measurement of 1100 individuals of 4 game * strain.
Animal
Day of
disease
Number
measured
Length in microns
Average
Maximum
Minimum
Monkey
7 1
7
2 5
H *79
32*9
17*0
)>
7 i
9
2 5
19-84
2 3*8
> 5*3
99
38
2 5
26*36
34* 2 5
19*0
tt
120
8
2 5
20*02
2 3*5
18*o
tt
120
11
2 5
21*9
19-15
> 7* 2 5
tt
120
>3
2 5
17*4
20*0
i5*o
it
>30
8
2 5
25-97
35*5
19*0
tt
130
11
2 5
22*05
30.5
16*25
tt
'99
5
2 5
22 *47
2 575
> 5*75
tt
>99
7
2 5
23*6
3 2 * 2 5
16-75
tt
201
7
2 5
2 3’4
31*0
>775
tt
201
8
2 5
21*62
2 5*5
> 7*5
tt
201
9
2 5
19-58
2175
'7-25
Dog, native
?
2 5
19*1
26*0
> 5*5
tt
262
5
2 5
21*69
2 575
.8-5
tt
262
7
2 5
19*13
2 3*5
> 3*5
tt
262
11
2 5
18-34
22 *5
16*25
Rabbit
79
11
2 5
20*02
29*0
>5* 2
»
249
9
2 5
l6*l8
>9*5
'375
tt
249
>3
2 5
22*29
32*0
> 5* 2 5
tt
249
>3
2 5
20*91
28-5
>575
Guinea-pig
2 5 >
10
2 5
20*87
33* 2 5
>5*25
2 5>
11
2 5
22*87
34*5
'575
tt
2 5 >
>3
2 5
23*11
33*75
15*0
tt
2 5 >
>5
2 5
23*5
3 2 * 2 5
> 4*75
tt
2 5 >
>7
2 5
2409
34* 2 5
>375
tt
2 5 >
21
2 5
21*67
29*75
> 4*5
Rat
81
>4
2 5
21*05
3>*5
16*o
tt
128
20
2 5
20*25
2 >*75
> 7*5
tt
128
22
2 5
20*3
2 3'75
16*o
tt
> 2 9
?
2 5
20*9
28*0
16*25
tt
>57
21
2 5
25*65
30-5
> 4*5
tt
>57
4 2
2 5
1927
2 >*5
1675
tt
>57
49
2 5
22*8
3 2 ’5
16*25
tt
>95
26
2 5
21*8
35*o
> 6*5
tt
>95
36
2 5
> 9*0
HS
17*0
tt
2 *3
>7
2 5
17*38
19*0
> 4*5
tt
2>3
26
2 5
22*31
34* 2 5
i 7*5
tt
221
7
2 5
18*91
23*0
16*5
tt
221
>4
2 5
21*91
35*5
1175
Mouse
176
9
2 5
20*13
26*5
> 7*5
tt
176
>4
2 5
20*99
26*5
16-75
tt
178
6
2 5
22*89
2 9*5
17.25
tt
178
7
2 5
21*6
27*0
16*5
1100
21*38
35*5
1175
220
Table 18.—Giving details of measurement of uoo individuals of ‘fly’ strain.
Animal
Day of
disease
Number
measured
Length in microns
Average
Maximum
Minimum
Monkey
96
7
2 5
2 5‘7
32-0
16-0
ji
96
8
2 5
24*8
33-5
.6-5
n
96
9
2 5
2 5'6
36-25
16-o
n
96
10
2 5
2 3'3
3075
*575
>>
96
11
2 5
22*6
31-0
*5-25
96
*4
2 5
20-3
2 3*5
16-5
* *4
2 7
2 5
22*0
28-0
16-25
1 >4
3 2
2 5
20*9
2 5‘ 2 5
18-0
1 14
4 *
2 5
20-8
3075
15-25
210
8
2 5
24-66
30-56
* 7*5
210
10
2 5
20-29
2 3* 2 5
18-25
2 *7
9
2 5
26-03
3075
22-0
u
3*6
9
2 5
24-69
3 2 * 2 5
* 7*5
Dog
2 35
5
2 5
26-7
33-0
19-0
jj
2 35
7
2 5
21-4
28-0
19-0
2 35
9
2 5
20-0
28-0
18*25
»
2 35
*3
2 5
20*0
21-25
18-5
Rabbit
2 45
7
2 5
2 3’5
2 9*5
16-5
2 45
8
2 5
20-0
28-0
* 4*5
2 45
9
2 5
>* 7 S
2 775
16-25
2 45
>3
2 5
22-84
30-0
17-0
Guinea-pig 246
*3
2 5
19-87
2 3‘5
167
246
>5
2 5
20-88
26-0
16-25
j>
246
18
2 5
17-63
2 **5
13-0
5 j
246
*9
2 9
19-0
2 7* 2 5
1 6*5
246
20
2 5
'*•95
2 5* 2 5
14-25
246
21
2 5
21-0
27-5
15-25
Rat
io 3
4
2 5
2 4 ‘*
30*0
17-0
jj
103
5
2 5
20-3
30-0
16-5
103
8
2 5
18-8
3075
* 4*5
218
6
2 5
* 9*47
2 4*75
* 6*5
»
218
9
2 5
19-3
29-0
* 4*5
218
H
2 5
20-1
22 ’5
17-0
218
16
2 5
* 9*41
2675
16-o
218
18
2 5
22*0
3°*5
18-o
5?
229
6
2 5
2 4*55
29-5
19-0
JJ
22 9
8
2 5
21-09
29-25
17-0
229
9
2 5
19-5
2I 75
* 7*5
>>
229
...
*3
2 5
22 *3
2 9*5
* 8*5
229
*5
2 5
20-31
22 75
17-0
Mouse
2 47
4
2 5
23-1
2 9*5
* 9*5
>»
2 47
6
2 5
23-66
29-5
19-25
2 47
9
2 5
22*6
34 *o
1*75
2 47
2 5
20*91
2 5* 2 5
>7-25
1100
21-67
36-25
13-0
221
Table 19.—Comparison of the measurements of the * human,' 4 game,* and 4 fly * strains.
Strain
Length in microns
Average
Maximum
Minimum
‘ Human *.
21-25
39 *°
13-25
‘ Game * .
21-38
35-5
m -75
•nr’ .
21-67
36 -iS
13-0
The similarity in the measurements is, perhaps, best appreciated
by a glance at the curves obtained by plotting out the distribution
of the various lengths of the parasites, expressed in percentages of
the total numbers measured (see Chart I).
A comparison of the percentages of * short and stumpy,’ ‘ inter¬
mediate ' and ‘ long ’ forms is also of interest.
Table 20. —Comparison of percentages of * short and stumpy,' ‘ intermediate,' and ‘ long ' forms of
the 4 human,* ' game,* and ' fly ' strains.
Strain
Short and stumpy
forms
Il-2tft
Intermediate forms
22-24/i
Long form»
2 S“ 39 M
1 Human'.
6478
15-98
I 9 -I 4
4 Game' .
62-87
15-34
21-56
4 Fly' .
58-68
18*81
22*41
2. 'Pathogenicity
The pathogenicity of the three strains is synopsised in Table 21,
p. 223. A glance at this table will show how remarkably the three
strains agree in this respect.
Tablx 21.—Comparison of the pathogenicity of the * human,* 1 game ’ and ‘ fly * strains of T. rbodesicnse .
223
H
(A
►*
£
Duration
days
7—60
Average 24
186
26
19—39
Average 29
47 — 1 *7
Average 74
(1 alive after 55
days)
9—58
Average 34
9 *
Incubation
days
II n *
> > > >
< < <! <1
* Gams * Strain
No.
used
12
1
1
5
4
10
1
Duration
days
7—63
Average 17
210
*5
14—32
Average 26
53—100
Average 72
11—87
Average 35
48—51
Average 44I
Incubation
days
4—ii
Average 6
7
S
4-8
Average 6
4—17
Average n
3 - 8
Average 5
4 — 5 f
Average 4$
No.
used
m m *r> v» W~> N
*5
g
«
Duration
1 d»y»
HR
OO - tv H
** On tv n m
If r - t| || tj II *
Incubation
days
3 — 9
Average 5
7
5
4— 6
Average 5
6—19
Average 11}
2-8
Average 5
4
6
as i
'H-
Animal
Monkey .
Ox.
Dog .
Rabbit .
1
Guinea-pig
Rat .
Mouse .
Genet .
224
SUMMARY
1. The human trypanosome ( T . rhodesiense) is distributed
widely throughout South Central Africa.
2. There is no essential difference between the clinical manifes¬
tations of the disease in man caused by T. rhodesiense and that due
to T . gambiense , except possibly the greater virulence of the former.
3. T . rhodesiense is transmitted in Rhodesia by Glossina
morsitans.
4. Approximately 3 5% of the flies may become permanently
infected and capable of transmitting the virus.
5. The period which elapses between the infecting feed of the
flies and the date on which they become infective varies from
eleven to twenty-five days in the Luangwa Valley.
6. Attempts carried out at laboratory temperature on the
Congo-Zambesi plateau during the cold season to transmit the
human trypanosome by means of Glossina morsitans were invariably
unsuccessful in spite of the fact that 680 flies ;were used in these
experiments.
7. The developmental cycle of T. rhodesiense in Glossina
morsitans is to a marked degree influenced by the temperature to
which the flies are subjected. High temperatures (75-85° F.)
favour the development of the parasite, whilst low temperatures
(6o°-70° F.) are unfavourable.
8. The first portion of the developmental cycle can proceed
at the lower temperatures, but for its completion the higher
temperatures are essential.
9. The parasites may persist in the fly at an incomplete stage
of their development for at least sixty days under unfavourable
climatic conditions.
10. These observations afford an adequate explanation of the
extremely long latent periods of trypanosomes in Glossina which
have occasionally been observed by various workers.
11. The relative humidity of the atmosphere has apparently no
influence on the development of the trypanosome in Glossina
morsitans .
225
12. Mechanical transmission does not occur if a period of
twenty-four hours has elapsed since the infecting meal.
13. Glossina morsitans , in nature, has been found to transmit
the human trypanosome.
14. The chief reservoir of the human trypanosome is the
antelope.
15. The results of examination for the human trypanosome of
the blood of a large number of monkeys, wild rats and mice were
invariably negative.
226
DESCRIPTION OF PLATE XVIII
Trypanosoma rhodesiense. Films fixed in alcohol and stained
with Giemsa. The figures were drawn with the aid of a camera
lucida at a magnification of 2,000 diameters.
Figs. 1-6. T. rhodesiense, * Human strain,’ obtained from a monkey
inoculated from a case of Sleeping Sickness.
Figs. 7-12. T. rhodesiense, ‘Fly strain,’ obtained by feeding
wild Glossina morsitans on healthy animals.
Figs. 13-18. T. rhodesiense, ‘Game strain,’ obtained from
game.
. Imuilx Tro/i M<u/. <v /hrtisilol.. I '<>/. Ill
Plate XVI,l 7
TRYPANOSOMA RHODESIENSE
A M Brookfield.del
127
SECTION II
TRYPANOSOMES OF GAME AND
DOMESTIC STOCK
BT
ALLAN KINGHORN
AND
WARRINGTON YORKE
Within the confines of the Luangwa Valley a numerous and
varied selection of game is found, despite the fact that Glossina
morsitans is everywhere abundant, but, on the contrary, due
doubtless to the presence of these insects, domestic stock is
extremely scarce, and in many districts non-existent. A few goats
are occasionally found, but some evidence exists to show that these
animals are not so insusceptible to trypanosomiasis, under natural
conditions, as is locally supposed. Dogs are very seldom seen,
and the natives themselves recognise the impossibility of keeping
them in the midst of ‘ fly.’ Cattle were seen in one village only.
At Ngoa, on the Congo-Zambesi watershed, game is abundant
at certain seasons of the year, and goats and dogs are kept in many
of the villages. Although tsetse flies are commonly seen near some
of these, such domestic stock animals as exist appear to thrive,
though trypanosomes were occasionally found in goats.
(a) METHODS
In our experience, trypanosomes are more readily detected in a
buck’s blood by the examination of thin, stained smears than by
that of fresh preparations. Except under unusual circum¬
stances, from one to two hours elapsed after an animal had been
shot before its heart reached the laboratory, and this, together with
the great heat, had a very deleterious effect cm the parasites,
destroying their motility, and permitting degenerative changes to
occur. In several instances in which fresh preparations, made
under these conditions, were examined, as well as blood smears
228
made in the field and afterwards stained, no trypanosomes were
found in the fresh blood, whereas they were present in the
permanent preparations.
In making smears, it was found advisable to cut the animal’s
throat immediately it had been shot, and to obtain the blood from
one of the arteries. It is claimed that such films have the following
advantages: —
1. Clean, uncontaminated preparations are obtained.
2. The trypanosomes have no opportunity of degenerating,
and thus stain more sharply.
3. The preparations are permanent.
4. The parasites can be identified more easily in stained, than
in fresh preparations. In this particular the examination of thin
films has an obvious advantage over that of thick films.
The preparations were dried in the air, fixed in absolute
alcohol, and stained with Giemsa.
Owing to the impossibility of obtaining clean sheep and goats,
all the game inoculations were made into monkeys and rats, the
amount of blood used varying from 1-10 c.cm. It is recommended
that, when possible, sheep and goats be used as well, since they are
susceptible to most of the pathogenic trypanosomes, whereas
animals such as dogs, monkeys and rats are not. This is an
important consideration when dealing with such parasites as
T. vivax and T. nanum.
(b) EXAMINATION OF GAME AT NAWALIA AND NGOA
A total of 127 head of game, comprising nineteen genera, was
examined at Nawalia, and trypanosomes were found by direct
examination, by inoculation, or by both methods, in thirty-three.
At Ngoa, 124 buck, belonging to sixteen genera, were examined,
and trypanosomes were found in twenty-one—a percentage of 16*9.
Details are given in Tables 22 and 23.
It will be seen that parasites were found at Nawalia by direct
examination in twenty-six cases, a percentage of 20*4, while at
Ngoa trypanosomes were found in the peripheral blood of only
sixteen buck—13 0%. These are high figures for single observa¬
tions, and it is probable that had several preparations from each
229
Table 22.—Results of examination of game for trypanosomes at Nawalia.
Animal
Number
examined
Number
in which
trypano¬
somes
were
found
in buck's
blood
Number
inocula¬
tions
made
Number
positive
inocula¬
tions
in which
parasites
were seen
in buck's
blood
Number
positive
inocula¬
tions in
which no
parasites
were seen
in buck's
blood
Total
number
buck
found
infected
by
examina¬
tion and
inocula¬
tions
1. Elephant
1
0
1
0
0
0
2. Rhinoceros ...
1
0
1
0
0
0
3. Hippopotamus
1
0
0
0
0
0
4. Zebra
5
0
3
0
0
0
5. Roan.
8
.
2
0
0
1
6. Wildebeest ...
2
0
1
0
0
0
7. Kudu
7
3
3
1
1
4
8. Hartebeest ...
6
0
1
0
1
1
9. Waterbuck .
28
16
H
5
1
*7
ic. Puku.
10
1
6
0
0
1
it. Mpala.
29
1
*3
1
1
2
12. Bushbuck .
9
4
6
1
2
6
13. Bushpig .
4
0
1
0
0
0
14 Warthog .
9
0
3
0
1
t
15. Lion.
2
0
0
0
0
0
16. Hunting dog
1
0
1
0
0
0
17. Giant rat .
1
0
0
0
0
0
18. Genet .
2
0
0
0
0
0
19. Squirrel .
1
0
0
0
0
0
127
26
56
8
7
33
230
Tablk 23. —Results of examination of game for trypanosomes at Ngoa.
Animal
Number
examined
Number
in which
trypano¬
somes
were
found
in buck's
blood
Number
inocula¬
tions
made
Number
positive
inocula¬
tions
in which
parasites
were seen
in buck's
blood
Number
positive
inocula¬
tions in
which no
parasites
were seen
in buck's
blood
Total
number
buck
found
infected
by
examina¬
tion and
inocula¬
tions
t. Rhinoceros .
6
0
3
0
0
0
2. Zebra .
*7
0
5
0
0
0
3. Buffalo .
6
0
3
0
0
0
4. Eland .
*5
0
12
0
4
4
5. Roan.
5
0
3
0
1
1
6. Hartebeest .
8
0
4
0
0
0
7. Waterbuck .
2 7
12
*5
3
0
12
8. Puku.
8
.
6
0
0
1
9. Sitatunga .
2
1
0
0
0
1
10. Duiker .
9
2
4
0
0
2
11. Klipspringer.
2
0
1
0
0
0
12. Warthog .
12
0
3
0
0
0
13. Hyaena .
2
0
1
0
0
0
14. Caracal .
2
0
0
0
0
0
15. Galago .
1
0
0
0
0
0
16. Reedbuck .
2
0
0
0
0
0
124
16
60
3
5
21
buck been searched, the percentage of successes would have been
much greater. In several instances, only a single trypanosome was
found in a film covering the greater part of a slide, and this after
a very careful examination extending over two hours.
A more accurate estimate of the percentage of animals
harbouring trypanosomes is afforded by considering only those from
231
which inoculations were made. An analysis of these gives the
following figures: —
Nawalia. Ngoa.
Number of inoculations made . 56 ... 60
Number of positive inoculations in which
parasites were found in buck’s blood... 8 ... 3
Number of positive inoculations in which
no parasites were found in buck’s blood 7 ... 5
Number of negative inoculations in which
parasites were found in buck’s blood ... 6 ... 6
Total number found infected . 21 ... 14
These figures show that at least 37 5% (Nawalia) and 23 3 %
(Ngoa) of the local fauna were infected with trypanosomes.
Both T. vivax and T. nanum have been found in game, and to
both these species monkeys and rats are refractory, so that no
conclusions can be drawn regarding the presence or absence of these
trypanosomes in animals in which parasites were not found in the
blood smears. Had sheep and goats been available for inocula¬
tion, it is probable that many more buck would have been shown to
harbour the two organisms in question. As a conservative estimate,
the percentage of game actually infected with trypanosomes in the
vicinity of Nawalia might be placed at 50, and at Ngoa 35.
A further point which is brought out in the tables is that
different species of buck appear to vary widely in their
susceptibility. Amongst the commoner varieties, trypanosomes
were never found either by direct examination, or by inoculation in
zebra, buffalo, wildebeest-and bushpig, and only rarely in roan,
hartebeest, puku, mpala and warthog. Waterbuck, eland, bush-
buck and kudu were the species found to be most heavily infected.
To a certain extent, perhaps, these differences may be accounted
for by the habitats affected by the various species of game. Kudu
apd bush buck, and waterbuck to a lesser extent, are usually found
in thick cover from which they seldom emerge, and where they
are more constantly exposed to the bites of tsetse flies. Mpala,
puku and wildebeest are usually found in open country, frequently
remaining for the greater part of the day on wide, bare plains, and
here the flies are less noticeable than in the bush. Specific
Tablk 24.— Percentages of various species of game found infected with trypanosomes at Nawalia.
Animal
Number examined
Percentage
harbouring
trypanosomes
Bushbuck .
9
66-6
Watcrbuck .
28
607
Kudu . .
7
57 '.
Hartcbcest .
6
16 6
Roan .
8
12’ S
Warthog
9
»•«
Puku .
10
10*0
Mpala ...
2 9
69
1
Tabu 25. —Percentages of various species of game found infected with trypanosomes at Ngoa.
Animal
Number examined
Percentage
harbouring
t rypanosomes
Sitatunga .
2
5 °
Waterbuck .
27
44*4
Eland.
*5
266
Duiker.
9
22-2
Roan .
5
20
Puku .
8
12-5
differences in the amount of immunity enjoyed by buck arc
probably, however, of much greater importance.
In Tables 26 and 27 are given the species of trypanosomes
occurring in each animal in which parasites were found. In
compiling the tables, information obtained from the result of
inoculations, where these were made, has been utilised. This
enables a differentiation to be made between such parasites
as T. pecorum and T. nanum , which are morphologically
indistinguishable. T. vivax has a characteristic morphology, and
can thus be identified in blood smears without difficulty.
*3 3
Table 26. —Trypanosomes found in game at Nawalia.
Animal
Trypanosomes found in
peripheral blood
Trypanosomes isolated by
inoculation into monkeys
and rats
Diagnosis
Bushbuck
1
Negative
7 . pecorum
7 . pecorum
2
7 . pc cor urn or 7 . nanum
Negative
7 . nanum
*»
3
T. multiforme , sp. nov.
7 . multiformc , sp. nov.
7 . mult t for me , sp. nov.
n
4
T. pecorum or 7 . nanum
No inoculation
7 . pecorum or 7 . nanum
r
5
7 . pecorum or T. nanum
V
7 . pecorum or 7 . nanum
o
6
Negative
7 . rbodesiense
7 . rbodesiense
Waterbuck
1
7 . pecorum or T. nanum
7 . pecorum
7 . pecorum
?»
2
7 . pecorum or 7 . nanum
7. pecorum and
7 . rbodesiense
7 . pecorum and
7 . rbodesiense
i'
3
7 . pecorum or T. nanum
and T. vivax
Negative
7 . nanum and 7 . vivax
0
4
7 . pecorum or T. nanum
7 . nanum
0
5
Negative
7 . pecorum
7 . pecorum
»
6
7 . vivo*
Negative
7 . vivax
7
7 . vivax
7 . vivax
??
8
7 . r bode tie me
7 . rbodesiense
7 . rbodesiense
Jl
9
T. pecorum or 7 . nanum
and 7 . vivax
Negative
7 . and 7 . vivax
10
7 . pecorum or T. nanum
No inoculation
7 . pecorum or 7 . nanum
11
(?) 7 . rbodesiense
Animal died day after
inoculation
(?) 7 . rbodesiense
T?
12
7 . rbodesiense
7 . rbodesiense and
7 . pecorum
7 . rbodesiense and
7 . pecorum
f J
n
7 . vivax
No inoculation
7 . vivax
??
m
7 . rbodesiense and
7 . vivax
7 . rbodesiense
7 . rbodesiense and
7 . vivax
»»
>5
7 . vivax
No inoculation
7 . vivax
»»
16
(?) 7 . rbodesiense and
7 . vivax
11
(?) 7 . rbodesiense and
7 . vivax
n
*7
(?) 7 . rbodesiense
»
(?) 7 . rbodesiense
Kudu
1
Negative
7 . pecorum
7 . pecorum
>»
2
7 . pecorum or T. nanum
No inoculation
7 . pecorum or 7 . nanum
3
7 . pecorum or T. nanum
7 . pecorum
7 . pecorum
4
T. pecorum or 7 . nanum
No inoculation
7 . pecorum or 7 . nanum
Roan
1
7 . pecorum or T. nanum
»
7 . pecorum or 7 . nanum
Warthog
1
Negative
7 . rbodesiense
7 . rbodesiense
Puku
1
T. vivax
No inoculation
7 . vivax
Mpala
1
Negative
7 . rbodesiense
7 . rbodesiense
»?
2
7 . pecorum or T. nasMsi
7 . pecorum or
7 . rbodesiense
7 . pecorum and
7 . rbodesiense
Hartebeest
1
Negative
7 . rbodesiense
7 . rbodesiense
234
Table 27.—Trypanosomes found in game at Ngoa.
Animal
Trypanosomes found in
peripheral blood
Trypanosomes isolated by
inoculations into monkeys
and rats
Diagnosis
Waterbuck
1
T. vivax
No inoculation
7. vivax
11
2
7. vivax
»
7. vivax
n
3
7. vivax
17
7. vivojc
n
4
7. vivax
Negative
7. vivax
»
5
T. vivax
11
7. vivax
M
6
7. vivax
7. rbodesiense
7. two* and
7. rbodesiense
11
7
7 vivax
7. rbodtsiense
7. vivax and 7. rbodesiense
»»
8
7. vivax
Negative
7. vivax
11
9
7. vivax
7. pecorum
7. vivax and
7. pecorum
11
10
7. vivax
Negative
7. vivax
»»
11
7. vivax
No inoculation
T. vivax
11
12
7. vivax
>s
7. vtvsx
Eland
1
Negative
7. pecorum
7. pecorum
11
2
11
7. pecorum
7. pecorum
it
3
ii
7. pecorum
7. pecorum
it
4
11
7. pecorum
7. pecorum
Roan
1
11
7. pecorum
7. pecorum
Puku
1
7. vivax
Negative
7. vivax
Sitatunga
1
7. tragelapbi , sp. nov.
No inoculation
7. tragelapbi) ip. nov.
Duiker
•
7. vivax
Negative
7. vivax
n
2
7. pecorum or 7. nanum
No inoculation
7. pecorum or 7.
As would be expected, double infections in game are not
uncommon, and several instances of this are recorded in the
tables.
No data exist as to the ultimate effect of infection on game.
All the animals which were shot appeared to be in perfect
condition, and presented no objective signs of disease.. Whether
235
or not buck succumb to trypanosomiasis it is impossible to say, but
as they have increased steadily since rinderpest swept through the
country, it may be assumed that their tolerance to trypanosomes is
very great.
(e) EXAMINATION OF DOMESTIC STOCK
The domestic animals examined, and the species of trypano¬
somes found in them, are given in Tables 28 and 29.
Tael i 28 .—Examination of domestic stock for trypanosomes at Nawalia.
Animal
Trypanosomes found in
peripheral blood
Trypanosomes isolated by
inoculation into monkeys
and rats
Diagnosis
Cow
.
T. pecorum or 7. nanum
No inoculation
7. pecorum or 7. nanum
.
7. pecorum or 7. nanum
”
7. pecorum or 7. nanum
Goat
- 39
7. vivax
Negative
7. vivax
... 94
7. vivax and 7. nanum
or 7. pecorum
it
7. vivax and 7. nanum
>»
... 202
7. pecorum at 7. nanum
Negative
7. nanum
»
... 258
7. vivax
>»
7. vivax
Dog
.
7. rbodesiense
7. rbodesiense
7. rbodesiense
J»
.
7. pecorum
7. pecorum
7. pecorum
t>
.
7. pecorum
No inoculation
7. pecorum
tt
.
7. pecorum
»t
7. pecorum
tt
.
7. sp. (montgomeryi ?)
Negative
7. sp. ( montgomeryi ?)
Table 29. —Examination of domestic stock for trypanosomes at Ngoa.
Animal
Trypanosomes found in
peripheral blood
Trypanosomes isolated by
inoculation into monkeys
and rats
Diagnosis
Goat ... 369
No inoculation
7. nanum or 7. pecorum
» - 375
7. vivax and 7. nanum
or 7. pecorum
>t
7. vivax and 7. ssssm
or 7. pecorum
- 37 *
7. vivax and 7. nanum
or 7. pecorum
7. pecorum
7. vivax and 7. pecorum
236
The only native village in which cattle were found was
Kambwiri’s, some forty miles south-west of Nawalia. At present
there are only three head, all that are left of a big herd which
existed there some four or five years ago. Two of the three
appeared to be in good condition when seen, but the headman of
the village fully expected to lose them within a few months. The
third beast was obviously ill. The cow in which trypanosomes
were found at Fort Jameson was bred on the Government Farm,
and had never been beyond the limits of the township. Tsetse
have never been seen within some miles of the place, but Stomoxys
is abundant in the kraals, and at certain seasons of the year various
species of Tabatiidae are common.
In several of the villages on the main road from Nawalia to
Fort Jameson, a number of goats were found at the end of August,
1911, and again at the beginning of April, 1912, but at the end of
that month not a single animal was alive. Glossina morsitans was
found around all these villages. The four goats mentioned in
Table 28 were under observation at Nawalia for a considerable
length of time. During this period, parasites were found in the
peripheral blood only at rare intervals. Two were rather thin, but
not markedly so, and, apart from this, there were no signs of
disease. Goat No. 258 was examined at frequent intervals for two
months before parasites were first found, while in the others,
trypanosomes were seen on the first occasion. Nos. 39 and 258,
after having been under observation for nine and four months
respectively, died on the road when the Commission left Nawalia,
most probably from being over-driven. The other two are still
alive, seven and four months after the diagnosis was made.
The dog in which T. rhodestense was found came from a
village just on the Nyasaland border. The natives said that it had
not been out of the village for over a year previously. As the
disease runs an extremely acute course in these animals, there can
be no doubt that the dog was infected locally.
237
( d) EXAMINATION OF SMALL VERMIN
It has been suggested that the small vermin might also act as
reservoirs of trypanosomiasis. It must be remembered, however,
that many of the small vermin of Tropical Africa are nocturnal,
and are, therefore, not subjected to the same extent as are the big
game to the bites of GL morsitans . At Nawalia and at Ngoa we
examined in all 142 wild rats, 15 wild mice, 1 wild rabbit, 1 giant
rat, 1 squirrel, 1 galago, and 2 genet; the results were uniformly
negative. Furthermore, it might be remarked that there is no
evidence to show that the small vermin are tolerant of the human
trypanosome as are the big game. In those which we infected
experimentally the disease ran an acute course, and the animals
died. If this be the case with the majority of the small vermin
they cannot have the same significance as reservoirs of the virus as
have the big game, which can probably harbour the parasite for long
periods without exhibiting signs of disease.
Not a single case of infection with trypanosomes was found in
the 256 monkeys (Cercofithecus pygerythus) examined by us,
although infection with filaria and Plasmodium kochi was common.
The probable explanation of this is that the monkeys during the
daytime catch the tsetse fly before the insects have time to feed on
them, whereas, on the other hand, they are frequently bitten by
mosquitos whilst they are asleep at night. Moreover, it must be
remembered that in these animals infection with the human trypano¬
some runs an acute course, and those animals which contract the
disease quickly succumb.
SUMMARY
Trypanosomes are of frequent occurrence in game and domestic
stock in North Eastern Rhodesia. As a conservative estimate the
percentage of big game infected with trypanosomes pathogenic to
man and domestic stock may at Nawalia (Luangwa Valley) be
placed at 50, and at Ngoa (Congo-Zambesi watershed) at 35.
At Nawalia six species of trypanosomes were isolated from game
and domestic stock, viz., T. rhodesiense, T. vivax, T. nanum ,
T. pecorum, T. montgomeryi, and T. multiforme; whilst at Ngoa
five species were found, viz., T. rhodesiense , T. vivax , T. nanum,
T. pecorum , and T. tragelaphi.
The results of examination of over 400 monkeys, wild rats and
mice were invariably negative.
239
SECTION III
TRYPANOSOMES FOUND IN WILD
GLOSSINA MORSITANS
BT
ALLAN KINGHORN
AND
WARRINGTON YORKE
During the sojourn of the Commission at Nawalia and at Ngoa,
experiments were undertaken with the object of ascertaining the
species of trypanosomes transmitted, in nature, by Glossina
morsitans, Westw. The flies, as they were brought to the
laboratory, were fed on clean monkeys, which were the only
animals available for the purpose. Unfortunately, owing to the
lack of healthy goats and sheep, no definite conclusions can be
drawn as to whether the fly was infected with such species as
Trypanosoma vivax and Trypanosoma nanum , both of which are
of common occurrence in game and domestic stock.
At Nawalia, in the Luangwa Valley, freshly-caught Glossina
morsitans were fed on healthy monkeys from day to day. In all,
3,410 flies were fed in batches, as they were brought to the
laboratory, on thirty-three monkeys, but as five of the latter died
within two or three days, inferences can only be drawn as to the
infectivity of the 3,202 flies fed on the remaining twenty-nine
animals. Details of the experiments are given in Table 30.
It will be seen from the table that three species of trypanosomes
were isolated, namely, Trypanosoma rhodesiense , Trypanosoma
pecorum, and a third, hitherto undescribed parasite, for which we
propose the name Trypanosoma ignotum.
240
Tablf. 30 .—Remit of feeding freshly-caught Glossina morsitans on healthy monkeys at Nawalia.
Number
of
Date
Number
of
Result
Trvpanosom .*i isolated
Experiment
—--
flies fed
—
1
21.6.11
18
Negative
4
57 **
128
Infection
7. ignotum, sp. nov.
7
19.7.1!
97
Negative
28
28.7 11
160
Infection
7. ignotum , sp . nov.
35
7.8.11
*93
n
7. ignotum. sp. nov.
95
30.10.11
90
Negative
96
30.10.11
82
Infection
7. rbodesiense
121
14.11.11
98
Negative
*43
4.12.11
103
100
6.12.1 1
*05
Infection
? 7 . rbodesiense , animal died day after
2*5
13.1.12
4 *
becoming infected
7. ignotum , sp. nov.
210
7-12.1.12
269
7. rbodesiense
2*7
16.1.12
200
n
7. rbodesiense
224
23.1.12
28
Negative
259
13.2.12
104
n
3*6
20.3.12
101
Infection
7. rbodesiense
3*7
,
2 1.3.12
11 2
7. pecorum
326
25 - 3-*2
93
Negative
329
26.3.I2
* 3 °
Infection
7. ignotum , sp. nov.
33 °
27.3.12
*37
7. ignotum , sp. nov.
333
28 . 3.12
74
»
7. rbodesiense
334
29 . 3 .I 2
6 7
Negative
336
30 . 3.12
109
Infection
7. ignotum y sp. nov.
34 °
1 . 4.12
108
7. ignotum , sp. nov.
342
2 . 4.12
52
Negative
343
34*2
85
Infection
7. rbodesiense and 7. ignotum , sp.
348
4.4.12
90
Negative
353
6.4.12
*37
Infection
7. pecorum and 7. ignotum , sp. nov.
94
15.9.11
*94
_
Goat; subinoculated monkeys and rats
did not become infected
241
An analysis shows that of the 3,202 flies used, at least nineteen
were capable of infecting monkeys. This figure is based on the
assumption that, with the exception of Experiments Nos. 343
and 353, each batch contained but a single infective fly. In each
of these experiments it is highly probable that at least two infective
flies wereapresent, as in the former both Trypanosoma rhodesiense
and Trypanosoma ignotum, sp. nov., were found in the monkey’s
blood, and in the latter both Trypanosoma pecorum and
Trypanosoma ignotum , sp. nov. The percentage of flies infected
with each of the three trypanosomes is given in tabular form.
Table 31. —Proportion of wild Glossina morsitans infected with 7 . rhodesiense , 7 . pecorum and
7. ignotum , sp. nov.
Species
Number
of
flies fed
Number
of
infections
Ratio of infected
to non-infected
flies
T. ignotum .
3,008
10
0
0
7. rhodesiense .
3,202
6 *
1 : 534
T. pecorum .
3,202
2
I : 1600
In our second interim report* one experiment in which
T. rhodesiense was obtained was inadvertently omitted, so that this
parasite was isolated in six, instead of five instances. In addition,
Monkey No. 100 became infected four days after the flies had been
fed on it. As the animal died the same day, we were unable to
decide the species of trypanosome present, but from the short
inoculation it is highly probable that the parasite was
T. rhodesiense. On this assumption the ratio of infected to
non-infected flies would be 1 to 455.
At Ngoa, on the Congo-Zambesi watershed, 5,250 freshly-
caught Glossina morsitans were fed in batches on forty monkeys
and two goats. Details are given in Table 32.
• Kinghom, A., and Yorke, W. A Further Report on the Transmission of Human
Trypanosome* bv Glossina morsitans , Annals of Tropical Medicine and Parasitology, 1912 ,
Voi vi, P . 269/
242
Table 32.—Results of feeding freshly-caught Cletsina morsitans on healthy monkeys at Ngoa.
Number
of
Experiment
Date
Number
of
flies fed
Result
Trypanosomes isolated
2 goats
May
248
Negative
Subinoculated motkeys did not become
infected with 7. rbodesienst or
7. peeontm
366
23.4.12
223
Infection
7. ignotum
367
23.4.12
212
»
T. ignotum
368
23.4.12
211
»»
T. ignotum
377
4.512
130
Negative
379
6.5.12
121
Infection
T. ignotum
395
16.5.12
*24
Negative
7. ignotum
404
28.5.12
146
Infection
7. rbodesienst and 7. ignotum
405
29-5*2
I09
»»
7. ignotum
4 * 2
1.6.12
90
Negative
421
4.6.12
* 5 *
Infection
7. ignotum
423
6.6.12
166
7. ignotum
429
11.6.12
92
Negative
43 o
12.6.12
120
»>
436
14.6.12
*45
439
17.6.12
136
n
444
19.6.12
*59
n
446
21.6.12
85
>1
456
24.6.12
120
11
4 60
25.6.12
*22
*»
461
26.6.12
Il6
Infection
7. rbodesienst
464
28.6.12
I IO
Negative
465
28.6.12
78
466
29.6.12
84
»?
469
1*7*12
Il8
Infection
7 ignotum
47 *
2.7.12
64
Negative
476
4.7.12
7°
»
__
_ . 1
_ . ..
_
,
Table 32. — continued.
Number
of
Experiment
Date
Number
of
flies fed
Result
Trypanosomes isolated
4*1
57* 2
80
Negative
485
6.7.12
96
487
8.7.12
162
Infection
T. rhodesiense
502
11.7.12
104
»»
T. ignotum and T. pecorum
5°3
12.7.12
168
Negative
506
13.7.1 2
121
5 *i
* 57 -* 2
84
Infection
T. ignotum
5*4
16.7.12
,2 3
Negative
5*5
1 7.7.1 2
140
Infection
7 . ignotum
518
19.7.12
*35
»
T. ignotum
5 21
19.7.12
118
ji 1
T. ignotum
5 2 4
20.7.12
160
>5
T. rhodesiense and T. ignotum
559
2.8.12
118
Negative
560
3.8.12
93
It will be seen from Table 33 that T. rhodesiense, T. pecorum
and T. ignotum were isolated from the plateau flies in the
proportion of 1 in 312, 1 in 1312, and 1 in 5250 respectively.
Table 33. —Proportion of wild Glossina morsitans infected with T. ignotum , T. rhodesiense and
T. pecorum.
Species
Number
of
flies fed
Number
of
infections
Ratio of infected
to non-infected
flies
7 . ignotum .
5,000
16
1 : 312
T. rhodesiense .
5 i 2 5 °
4
1 : 1312
T. pecorum .j
5 ’ 2 5 °
! 1
!
* • 5 2 5 °
SUMMARY
T. rkodesiense , T. ignotum and T. pecorum are transmitted by
Olossina morsitans in nature, and were obtained by feeding wild
freshly-caught Glossina morsitans on healthy monkeys.
*45
SECTION IV
DESCRIPTION OF TRYPANOSOMES
FOUND
BY
ALLAN KINGHORN
AND
WARRINGTON YORKE
1. TRYPANOSOMA RHODESIENSE (PI. XVIII)
This parasite has been fully dealt with in a previous section,
and requires, therefore, no further description. It was isolated
from all the cases of Sleeping Sickness, sixteen in number, observed
by the Commission. At Nawalia it was found in four waterbuck,
two mpala, one hartebeest, one bushbuck and one warthog—
16% of the game from which inoculations were made—and from
one native dog. Parasites resembling T. rhodesiense were found
in blood films made from three other waterbuck, from which no
sub-inoculations were made. At least six, and possibly seven, of
3,202 freshly-caught Glossina morsitans were found capable of
transmitting this organism. At Ngoa the trypanosome was isolated
from two waterbuck, and from four of 5,250 freshly-caught tsetse
flies.
2. TRYPANOSOMA VIVAX (PI. XIX, figs. 1-8)
At Nawalia this organism was found in eight waterbuck, one
puku and three goats, and at Ngoa in twelve waterbuck, one puku,
one duiker and two goats.
MORPHOLOGY
{a) In fresh preparations it appears as a club-shaped organism,
characterised chiefly by the extraordinary rapidity with which it
moves across the field.
( b) In stained preparations it is seen to be more or less club-
shaped, with a long free flagellum. The greatest width is posterior
to the nucleus, which is situated about the middle of the body.
246
Table 34. —Measurements of * 7 . vivax.
Animal
Day of
disease
Number
measured
Length in microns
Average
Maximum
Minimum
Goat ...
39
i
25
24*35
26
22-5
39
i
25
23-67
26-25
2125
.
<✓»
00
t
25
2243
25*5
*975
n
258
?
25
22-74
25*5
21-25
,, ...
258
?
25
28-25
20-25
11
448
l 7
25
24-69
2825
1875
,y
448
18
25
24*5
2875
20-25
,,
448
19
25
23-22
26-5
19-25
200 !
2363
28-75
>875
Mickons
Chart 2. —Giving the curve representing the distribution, by percentages, in respect
of length, of Trypanosoma vivax.
247
The blepharoplast is large and rounded, and lies close to the
posterior extremity of the parasite. The undulating membrane is,
as a rule, very feebly developed, or absent.
The mean length of 200 trypanosomes was 23 6 n, the maximum
2875/», and the minimum 1875M (see Table 34). The greatest
width varied between 2 and 425/u, average 3‘2/x.
PATHOGENICITY
Inoculations were made into the
8 monkeys .
2 rabbits .
8 rats .
following animals: —
all remained negative.
both ,,
all ,,
I >
> I
TRANSMISSION
Owing to the fact that we were unable to obtain a number of
clean goats, we could not ascertain definitely that Glossina
morsitans transmitted this trypanosome in nature. Nevertheless,
that this fly can transmit T. vivax is shown by the following two
experiments. The wild Glossina morsitans used for breeding
purposes were fed regularly for over two months on goats which
were naturally infected with T. vivax and T. fecorum. These
goats were obtained from Ngoa, in the vicinity of which tsetse
flies were abundant.
Experiment I
On July 3rd, forty-eight of the breeding flies, which had
previously been starved for five days, were fed on a young, healthy
goat. The goat’s blood had been examined regularly for ten days
before the commencement of the experiment, and no parasites
were found. Ten days later the animal became infected with
Trypanosoma vivax .
Experiment 2
Three other flies were fed on a healthy kid on June 24th, and
twelve days later T. vivax appeared in the peripheral circulation.
The insects were dissected on the day after they had fed on the
goat, and trypanosomes were found in one only. The infection
248
was confined to the proboscis, in which the parasites were extremely
numerous and disposed in large rosettes.
Both these goats were brought, in mosquito-proof cages, from
a fly-free area.
3. TRYPANOSOMA NANUM (PI. XIX, figs. 9-16)
Found in the following animals at Nawalia:—One bushbuck,
three waterbuck and two goats. Possibly it was also present in two
other bushbuck, one other waterbuck, two kudu, a roan and two
cattle, but as no sub-inoculations were made, it was impossible to
differentiate it from Trypanosoma pecorum. At Ngoa the parasite
may have been present in a duiker and two goats, but in the absence
of sub-inoculations it could not be distinguished from T . pecorum .
MORPHOLOGY
(a) In fresh preparations it appears as a short, sluggish
organism. As a rule, it does not progress.
(b) In stained preparations it is found to be short, with a more
or less rounded posterior extremity from which the body tapers
forwards to the acute anterior end; the nucleus is placed at the
centre of the body. The blepharoplast is small, and is situated
near the posterior extremity. There is no free flagellum, and
the undulating membrane is absent, or, at most, very slightly
developed. The protoplasm is free of vacuoles and granules, in
general.
The mean length of 200 individuals was 14*3/1, the maximum
19/u, and the minimum io/i (see Table 35). The breadth, at the
level of the nucleus, varied from 1 to 2*25/1, the average being 1*5/1.
PATHOGENICITY
The following animals were inoculated: —
3 monkeys ... ... ... all remained negative.
1 rabbit . ,, ,,
3 rats ... ... ... ... ,, ,, ,,
Table 35.—Measurements of T. tianum.
Animal
Day of
disease
Number
measured
Length in microns
Average
Maximum
Minimum
Goat ... 202
Naturally
infected
25
15-2
19-0
12-25
„ ... 202
»>
25
*477
. 8-5
ii *5
„ ... 202
25
14.0
I7-0
ii *5
„ ... 202
25
14-61
18*0
ii *5
„ ... 202
25
13-99
I 7 -0
ii *5
,, ... 202
jj
25
* 3-51
18-o
io-5
n - 202
25
* 4*43
1875
| 10-0
„ ... 202
25
1
14*39
17-0
I 12-0
!
1 200
1436
19-0
I o-o
Microns
Chart 3. —Giving the curve representing the distribution, by percentages, in respect
of length, of Trypanosoma nanum.
DESCRIPTION OF PLATE XIX
Trypanosoma vivax and Trypanosoma nanum. Films fixed in
alcohol and stained with Giemsa. The figures were drawn with the
aid of a camera lucida at a magnification of 2,000 diameters.
Figs. 1-8. T. vivax.
Figs. 9-16. T. nanum.
. haulI s fro/). Med. d 1 ‘u'ttui/<)!,, 1of. I//
Plate XIX
TRYPANOSOMA VIVAX
A M Brookfield.del
TRYPANOSOMA NANUM
251
TRANSMISSION
The vector of T. nanum has not been determined with certainty.
4. TRYPANOSOMA PECORUM (PI. XX, figs. 1-8)
At Nawalia this parasite was found in one bushbuck, one
mpala, four waterbuck, two kudu, three dogs and one wild rat.
It was possibly present in two other bushbuck, a fifth waterbuck,
two other kudu, a roan and two cattle, but as mentioned before,
it could not be distinguished from T. nanum in the absence of
sub-inoculations. It was also isolated from 2 of 3,202 freshly-
caught Glossina morsitans.
At Ngoa it was obtained from one waterbuck, four eland, one
roan and one goat. It was possibly present in one duiker and two
other goats. It was also found in one wild tsetse fly.
MORPHOLOGY
(1 a ) In fresh preparations, this parasite resembles T. nanum very
closely. It is a short, sluggish organism, showing no degree of
translatory power.
(b) In stained preparations it appears as a short organism, with
an obtuse or rounded posterior extremity from which it tapers to
the attenuated anterior end. The nucleus is oval or rounded, and
situated near the middle of the body. The blepharoplast is small
and rounded, and lies close to the posterior extremity. The
undulating membrane, if present, is very feebly marked, and there
is no free flagellum. The general protoplasm stains uniformly,
and is devoid of granules and vacuoles.
The mean length of 500 individuals was 13*6/1, the maximum
19/1, and the minimum 9*5/1 (see Table 36). The breadth, at the
level of the nucleus, varied from 1 to 2*5/1, the average being 1*5/1.
PATHOGENICITY
A synopsis of the pathogenicity is given in Table 37, p. 253.
252
Table 36.—Measurements of T. pecorum.
Animal
Day of
disease
Number
measured
Length in micront
Average
Maximum
Minimum
Ox
393
11
25
*373
18*25
10*25
»
393
*4
25
12*92
* 5*5
975
Monkey
2
21
25
13*27
16*25
10*0
»»
9
9
25
.3-0
16*8
10-3
37
29
25
I +-47
1675
11*0
109
27
25
* 3*37
'675
io*5
«
109
36
25
■ 3'°9
*575
io*75
n
*23
28
25
I5*26
19*0
12*0
i»
3*7
43
25
I4*28
*75
11*0
»>
3*7
47
25
* 3*54
* 7*5
11*0
n
339
*4
25
15*61
*775
**75
Dog
274
(?) Naturally
infertrH
25
1409
•675
I 1*0
Guinea-pig
: 3 *
UUWV Ivu
21
25
12*33
'55
9*5
Rat
49
9
25
* 3*7
16*o
9*8
l6 3
20
25
13*26
*525
1075
163
23
25
* 3**3
16*25
*075
n
282
5
25
12*38
*475
9*5
»»
282
II
25
*375
16*o
! 10*25
Mou<e
11
12
25
*3'4
16*o
io*6
p
12
5
25
I 4‘ l
16*8
9*8
5°o
13*6
10*0
9*5
253
Microns
Chart 4.—Giving the curve representing the distribution, by percentages, in respect
of length, of Trypanosoma pecorum .
Tabu 37.—Pathogenicity of Trypanosoma pecorum.
Animal
1
No. inoculated
Incubation period
days
Duration
days
Monkey .
8-21, average
12*
23-225 (still alive)
Rabbit .
3
8-11, „
IO
66-215
Guinea-pig .
1
5
39
Rat .
14
3-14, average
64
6-32
Mouse
2
5
~ 7“75
Goat.
3
11-18, average
*5
53-58 (still alive)
Ox .
1
9
2 5 +
TRANSMISSION
T. pecoTum was obtained in two of twenty-eight experiments in
which freshly-caught Glossina morsitans were fed on clean monkeys
at Nawalia, and in one of forty experiments at Ngoa. In all,
therefore, 3 of 8,452 wild flies were found to be naturally infected.
At Ngoa batches of breeding flies which had fed for two to three
months on goats, naturally-infected with T. pecorum, were allowed
to feed on three healthy goats. That on which the first batch,
consisting of thirty flies, were fed became infected with the parasite
on the nth day of the experiment. Trypanosomes were found in
the blood of the second goat, on which eight flies were fed, on the
12th day. The third goat, on which three flies were fed, became
infected after an incubation period of eighteen days. The flies in
the last two groups were dissected the day after they had fed.
Trypanosomes were found in the gut and proboscis of a number,
but in no instance was an infection of the salivary glands observed.
These experiments afford satisfactory proof that Glossina
morsitans transmits the parasite in nature.
5. TRYPANOSOMA MULTIFORME, sp. nov.
(PI. XX, figs. 9-16)
This parasite was isolated from a bushbuck at Nawalia.
MORPHOLOGY
(a) In fresh preparations, the trypanosome is seen to be
markedly polymorphic, resembling in this particular Trypanosoma
rhodesiense. Trypanosoma gambiense and Trypanosoma pecaudi.
Short sluggishly-moving forms are seen, as well as long free-
flagellated, active ones. As a rule, the short varieties are more or
less stationary, while the long ones progress fairly rapidly. The
proportions of each of the forms varies widely, from day to day,
in any one animal.
(A) In stained preparations, every gradation between extremely
short, aflagellar forms, indistinguishable from T. pecorum, and
long, slender, free-flagellated parasites are seen. In general, the
255
trypanosome morphologically closely resembles T. gambiense or
T. brucei, except for the presence of occasional pecorum -like forms.
The mean length of 1,000 individuals was 2118/1, the maximum
33 5/1, and the minimum io'5/i. The curve (Chart 5, p. 259)
representing the distribution of the various lengths of the trypano¬
somes, expressed in percentages of the total number measured (1,000)
differs from that of T. gambiense in that the apex occurs at 17/1.
Table 38. —Measurements of T. multiforme , sp. nov.
Animal
Day of
disease
Number
measured
Length in microns
Average
Maximum
Minimum
Monkey
166
47
25
18-26
0
vO
*425
166
62
25
16-2
18* 5
*375
t?
166
78
25
* 5*34
180
12*0
tt
3*2
12
25
* 7-93
2475
*075
it
312
*3
25
t6-84
23*0
*375
11
3*2
*9
25
18*09
27*5
*575
11
3*2
42
25
19-65
28*0
*7*25
11
3*2
23
25
2473
3 0<2 5
130
11
3*2
44
25
17-48
28*0
13*0
11
360
9
25
2373
30*75
15*0
it
Jfo
>4
25
18-88
24*5
*475
11
360
*9
25
20*22
27*75
* 5*5
Rabbit
370
7
25
18*36
28*25
**75
»t
370
*5
25
24*1
1
29*75
16*25
tt
370
16
2?
20*55
29*0
13*25
it
370
81
25
23*49
2875
* 9*5
tt
4*4
5
25
25*82
33*5
14*0
tt
4*4
12
25
2576
31*0
* 5*5
tt
4*4
*3
25
23-42
30*0
12*0
tt
4*4
16
25
21*1
29*25
IO.5
256
Table 38.—T. multiforms, continued.
Animal
Day of
disease
Number
measured
Length in Microns
Average
Maxmum
Minimum
Rabbit 414
18
2 5
2 *-49
28*5
*375
„ 414
22
2 5
26*03
32*25
18*25
,1 4*4
7 *
2 5
26*41
3**5
20*0
». 4*4
7*
2 5
25-84
30*5
*7*25
v 4*4
79
2 5
238
29*5
12*0
„ 4*4
81
2 5
2 **44
28-75
15*0
Rat 219
7
25
2 * *75
31*25
2375
M 219
*3
2 5
2 479
32*0
16*25
v 219
3 2
2 5
20*32
26*25
*575
» 2 *9
4 2
2 5
18-54
26*0
14*0
» 219
43
2 5
20*0
30*0
14*25
» 219
44
2 5
l6-6l
23*25
1275
it 2 *9
45
2 5
16*08
2575
*275
>• 253
12
2 5
25-38
33 *°
*575
» 2 53
*3
2 5
22*1 I
33*25
*575
n 2 53
*9
2 5
21*46
30*25
*3*25
» 3 6 *
11
2 5
20*97
32*25
*7*25
» 36*
21
2 5
21*64
30*0
16*0
» 3 6 *
40
2 5
19*23
30*0
*5*25
>1 36*
53
2 5
*7*24
*9*5
12*5
1,000
21*18
33*5
j *°*5
257
Tabu 39.—Pathogenicity of Trypanosoma multiforme.
Animal
Incubation period
days
Duration
days
Remarks
Monkey
166
8
94
Tryps. last seen on 78th day.
'!
3*2
10
74
Tryps. last seen on 55th day.
11
360
7
.56
Tryps. last seen on 65th day.
11
455
4
Still alive noth
day
Tryps. last seen on 16th day.
Rabbit
370
6
* *9
Tryps. last seen on 112th day.
'i
4*4
—
—
Did not become infected.
5 after 2nd
inoculation
69
Tryps. present at time of death.
Guinea-pig
37 *
—
-
Did not become infected. Inocu¬
lated twice.
4*3
—
—
1 ' 11
1?
454
—
—
11 11
Rat
219
7
104
Tryps. last seen 44th day.
i»
2 53
12
*25
Tryps. last seen 86th day.
i?
361
10
67
Tryps. last seen 61 st day.
Ox
39 *
—
—
Did not become infected. Inocu¬
lated twice.
Goat
5 °*
—
—
DIAGNOSIS
This parasite is at once distinguished from T. fecaudt and
T. rhodesiense by the absence of posterior nuclear forms and by
its slight pathogenicity for laboratory animals. It more closely
resembles T. gambiense than any other known species, and is not
easily distinguished from this parasite. A study of the biometric
curves of the two trypanosomes shows certain differences, and if the
percentages of short, intermediate and long forms of each be
compared, as in Table 40, the difference is more clearly brought
out, the parasite in question exhibiting very few intermediate forms.
2 5 8
Table 40. —Companion of percentages of ‘short and stumpy/ ‘intermediate/ and ‘long ’forms
of T. multi forme and T. gambiense.
Short and stumpy
1
1 Intermediate
Long and slender
10—21 fi
| 22—24^
25 — 33 M
Trypanosoma multiforme
12-5
3**0
Trypanosoma gambiense
; 51-2
23-1
257
The extreme chronicity of the disease caused by this parasite in
laboratory animals is an additional point of distinction from
T. gambiense . We conclude, therefore, that the parasite is a new
species, and in view of its marked polymorphism we propose for it
the name Trypanosoma multiforme.
259
$ 3 OVM H 3 dy 9 4
4 .
26 o
DESCRIPTION OF PLATE XX
Trypanosoma pecorum and Trypanosoma multiforme. Films
fixed in alcohol and stained with Giemsa. The figures were drawn
with the aid of a camera lucida at a magnification of 2,000
diameters.
Figs. 1-8. T. pecorum.
Figs. 9-16. T. multiforme.
261
TRANSMISSION
The vector of T. multiforme is unknown.
6 . TRYPANOSOMA SP . (? MONTGOMERY!)
(PI. XXI, figs.9-16)
Found in one dog, which was obtained from a village in the
hills on the Nyasaland border.
MORPHOLOGY
(rf) In fresh preparations, the parasite appears as a broad,
stumpy organism, which shows no marked degree of translatory
power.
(b) In stained preparations, the trypanosome resembles at first
sight T. pecorum y but on closer examination it can be readily
distinguished from this parasite by its great breadth. The ratio of
the breadth to the length is 1 : 4*8. The posterior extremity is
subacute or rounded, the anterior attenuated. The greatest width
lies at the level of the nucleus, which is situated at the middle of
the body, or posterior to it. The blepharoplast is very large and
rounded, and is situated near the posterior extremity. Frequently
it lies at one edge of the trypanosome, and projects laterally as a
small excrescence. The undulating membrane is, as a rule, absent,
and when present is simple, and feebly developed. Occasionally
a short free flagellum, 1-2/* in length, is to be seen, but this is
generally absent. The cytoplasm often contains coarse granules
and vacuoles. The latter are most commonly seen in the posterior
portion of the body, whereas the granules may be scattered
generally throughout the protoplasm.
The average length of 200 individuals was 15*88/*, the maximum
20and the minimum 10/*. The breadth, at the level of the
nucleus, varied from 125-6*5/1, the average being 3*29/1.
PATHOGENICITY
One rat was sub-inoculated from the dog, and had not become
infected up to the 13th day afterwards, when it was accidentally
killed. Unfortunately the dog died some days previously, so that
the strain was lost.
262
Tapi.e 4 i. —Measurements of 7. montgomeryi.
Animal
Day of disease
Number
measured
Length in microns
Average
Maximum
Minimum
Dog 206 .
Naturally- 27.1.12
2 5
■s-s 6
18-25
12-5
infected
,, 206 .
,, 28.1.12
2 5
15-83
20*0
12*5
206 .
29.1.12
2 5
' 5-53
18-5
11 75
206 .
30.1.12
2 5
16-88
1 9* 2 5
14-0
.. 206 .
31.1.12
25
16-84
19-0
14-0
,, 206 .
.. 1.2.12
2 5
1 4 * 7 2
*775
ii *5
i
206 .
3.2.12 j
2 5
15-20
*775
10-0
., 206 .
,, 4.2.12
2 5
16-55
1875
* 3*5
200
,
15-88
20*0
10-0
Microns
Chart 6. —Giving the curve representing the distribution, by percentages, in respect of
length, of Trypanosoma montgomeryi.
263
TRANSMISSION
The vector of T . montgomeryi ( ?) is unknown.
DIAGNOSIS
It will be seen that, morphologically, this parasite differs widely
from T. pecorum. The average length is greater, 15*88/1, as
compared with 13*6/1. The most characteristic difference, however,
is the great width of the organism, which is, on an average,
2*2 times that of T. pecorum : average breadth 3*29/1, as compared
with 1*5/1. It appears to resemble most closely the parasite
described by Montgomery and Kinghorn* as the Ninamwenda
strain, and for which the name Trypanosoma montgomeryi was
proposed by Laveran. But in view of the fact that we were able
to examine material from one dog only, we cannot regard its
identity with this parasite as established.
7. TRYPANOSOMA IGNOTUM , sp. nov. (PL XXI, figs. 1-8)
This parasite was obtained by feeding with Glossina motsitans
on monkeys on ten occasions, and assuming that only one
fly was infective in each instance, the proportion of infective to
non-infective flies is 1 to 300, or 0*3 %.
MORPHOLOGY
(a) Fresh preparations. The parasite appears as a compara¬
tively short slender organism. It is fairly actively motile, but
exhibits no marked degree of transitory power.
(£) Stained preparations . The trypanosome is a slender
organism of moderate length. The posterior extremity is obtuse,
or bluntly rounded, while the anterior is effilated. The nucleus is
rounded or oval, and lies at the middle of the body. The
blepharoplast is small and rounded, and while usually situated
near the posterior extremity, may be separated from it by an
appreciable interval. It frequently projects from the edge of the
parasite, forming a bud-like excresence. The undulating membrane
is feebly developed, and a short free flagellum is only very
occasionally present. As a rule, the cytoplasm shows no granules
or vacuoles.
The average length of 500 individuals was 16*8/1, the maximum
23/i, and the minimum 12/1 (see Table 42).
•Montgomery and Kinghorn, Annals of Trop. Med. and Parasit., Vol. Ill, No. 2, 1909.
264
Table 42.—Measurements of T. ignotum.
Animal
Day of
disease
Number
measured
Length in microns
Average
Maximum
Minimum
Goat
45 *
*7
2 5
* 5*55
18*25
13*25
Monkey ...
1
8
2 5
16*67
20*25
* 3*5
n
22
10
2 5
* 7‘°4
20*25
1375
51
22
11
2 5
17*99
20*0
* 5-5
1)
...
36
6
2 5
16-55
20*5
13*0
11
230
9
2 5
17-38
23*0
* 3*5
11
...
250
11
2 5
16*52
19*0
*3*25
11 •**
363
9
2 5
17*1
20*0
12*0
»1
469
7
2 5
* 7*45
*975
*375
469
8
2 5
1 7 '* 9
*9*5
*3*0
>1
469
9
2 5
16*83
19*25
*375
15
518
12
2 5
16*99
19*0
*475
11
521
9
2 5
* 7**9
*975
*45
,,
5 21
10
2 5
16*87
20*25
* 4*5
Rabbit ...
44
37
2 5
16*9
* 9* 2 5
*375
„
45 2
18
2 5
167
20*25
*3*25
«
45 2
20
2 5
* 5*43
19*5
i2 *5
ii
45 2
21
2 5
* 5*93
2 o *75
14*0
it
...
497
*3
2 5
16*88
21*0
14*0
>1
...
497
2 5
2 5
16-93
19*5
* 3* 2 5
500
16*8
23*0
12*0
265
Microns
Chart 7.—Giving the curve representing the distribution, by percentages, in respect of
length, of Trypanosoma ignotum.
PATHOGENICITY
In monkeys, the virulence of Tryp. ignotum was found to be
very great. The disease is of a fulminating character, the parasites
increasing rapidly in number until the animal’s death. The
trypanosome is of equal virulence in those animals infected directly
by the bites of the tsetse flies and in those cases where the strain
was passed from monkey to monkey. The incubation period varied
from three to ten days, the average being seven, and death occurred
two or three days after the parasites appeared in the peripheral
blood.
It is interesting to note that on reaching the plateau three
monkeys failed to become infected when inoculated with the valley
strain of the parasite. At Nawalia, a failure was not recorded, and
it is difficult to understand those mentioned, more particularly as
the parasite is of frequent occurrence in the plateau flies, and
monkeys infected with this strain react in the same manner as those
infected by the valley strain.
Three rabbits were successfully infected with the strain by
sub-inoculations from a monkey. The incubation period varied
from 13 to 22 days, and the duration of the disease 66 to 106
(still alive).
Two guinea-pigs, twelve rats, four mice, an ox, a goat and a
dog were found to be refractory. Moreover, negative results were
obtained by feeding infective flies on rats.
Table 43.—Pathogenicity of Trypanosoma ignotum.
Animal
Number used
Incubation
days
Duration
days
Remarks
Monkey.
36
3—10
Average 7
5-16
Average 14
Three did not become
infected.
Rat .
12
—
—
None became infected.
Guinea-pig ...
2
—
—
Did not become infected.
Rabbit .
3
13—22
Average 17
16—106
Two alive after over 100
days
Mouse .
4
—
—
Did not become infected.
Ox .
1
—
—
Goat .
'
l 7
68
Still alive
Tryps. only seen once.
Monkey inoculated on
55th day did not
become infected.
Dog .
1
—
Did not become infected.
DIAGNOSIS
Morphologically the parasite appears to be distinct from any
hitherto described species. The graph (Chart 7) showing the distribu¬
tion of the trypanosomes in respect of length resembles very closely
that of Trypanosoma uniforme , but the parasites are at once
distinguished by the fact that whereas Trypanosoma uniforme is
267
invariably furnished with a free flagellum,* this, as mentioned above,
is of rare occurrence in Trypanosoma ignotum , sp. nov. Moreover,
the difference between the two trypanosomes is clearly demonstrated
by reaction of sub-inoculated animals. According to the Royal
Society Commission, Trypanosoma uniforme is innocuous to
monkeys, an observation which has been confirmed by Fraser and
Duke,t who record that they were unable to infect these animals by
sub-inoculation from game harbouring the parasite, although goats
were readily infected. However, the fact that a large number of
monkeys, and one rabbit, quickly succumbed to the disease indicates
clearly that the two parasites are not identical.
No information is at present available regarding the alternative
host of the trypanosome. Although the parasite has been isolated
from wild Glossina morsitans much more frequently than any other
trypanosome, it has never been found in game or domestic stock.
Nothing resembling it has been seen in the peripheral blood of any
animal examined in the Luangwa Valley and on the Congo-Zambesi
watershed. These include 250 wild animals (elephant, rhinoceros,
hippopotamus, buffalo, eland, zebra, wildebeest, roan, kudu,
hartebeest, true waterbuck, Crawshay*s waterbuck, puku, mpala,.
bushbuck, duiker, klipspringer, bushpig, warthog, lion, hunting
dog, caracal, galago, squirrel, genet, giant rat and rabbit),
35 domestic animals (cattle, goats and dogs), 256 monkeys, 142 wild
rats and 15 wild mice—making a total of 698. Eighty-six monkeys
have been sub-inoculated from game and domestic animals, and in
no instance has an infection with this trypanosome been observed.
In view of the fact that we have been unable to find the
vertebrate host, we propose to name the parasite Trypanosoma
ignotum.X
*Reports of Sleeping Sickness Commission of the Royal Society, No. XI, 1911, pp. 160-164.
tFrascr, Capt. A. D., and Duke, H. C. Bull. S.S. Bureau, Vol. IV, No. 36, April, 1912, pp.
151-152.
X On returning to England at the conclusion of the work of the Commission, we found that
the Royal Society Commission had published an account of a parasite which they called T. simiae.
This is almost certainly identical with T. ignotum. As the paper of the Royal Society Commission
appeared a few days before our paper on 7 . ignotum , this name must be dropped in favour of
7 . simiae , should the two eventually prove to be identical. (W.Y.)
DESCRIPTION OF PLATE XXI
Trypanosoma ignotum and Trypanosoma montgomeryi . Films
fixed in alcohol and stained with Giemsa. The figures were
drawn with the aid of a camera lucida at a magnification of
2,000 diameters.
Figs. i-8. T. ignotum.
Figs. 9-16. T. montgomeryi.
269
TRANSMISSION
In one experiment the infective fly was determined to be one of
a group of ten. These were then killed and dissected. Nine of
the flies were found to show no trypanosomes in the gut, proboscis,
salivary glands or sucking stomach, whereas in the tenth a heavy
infection of the proboscis was encountered. The gut, salivary
glands and sucking stomach were negative. This observation would
indicate that the development of the trypanosome, T. ignotum ,
occurs in the proboscis.
8. TRYPANOSOMA TRAGELAPHl , sp. nov. (PI. XXII)
This parasite was found in blood films made from a sitatunga
(Tragelaphus spekei) shot near Mpika.
MORPHOLOGY
It is at once distinguished from all known mammalian trypano¬
somes, with the exception of T. in gens, to which it bears a very
close general resemblance—so close, in fact, that in a previous report
we referred to it by this name.
It is a long, fairly broad trypanosome with a well-marked
undulating membrane, sometimes terminating in a short free
flagellum. The posterior extremity is effilated. The nucleus
appears as a broad band lying transversely across the centre portion
of the body, and stains faintly with Giemsa. The blepharoplast is
small but distinct, and is situated slightly posterior to the nucleus.
The length of the five specimens seen in the single film examined
were respectively 52*5, 53, 66, 70 and 72*5 microns, whilst the
breadth at the level of the nucleus was 6, 8*5, 5, 6 and 7 microns.
DIAGNOSIS
Whether or not this parasite is T . in gens, we are, from the small
amount of material available, unable to state with certainty. The
general resemblance of the two trypanosomes is striking, but if the
camera lucida drawings of this parasite be compared with those of
T. ingens published in the report of the Royal Society Commission,
270
it will be seen that the trypanosome in question is shorter and more
slender than T. ittgens , and that the undulating membrane is
broader and more pronounced. In the original description of
T. ingens the length of five specimens are given as 72, 77, 82, 88
and 122 microns. The breadth is stated to be from 7 to 10 microns.
It will be thus seen that T. ingens is on an average considerably
longer than the parasite in question. The average length of the
specimens of T. in gens measures 88 microns, whereas that of the
specimens of this parasite is only 63 microns. We propose
T. tragelaphi as a name for this parasite.
TRANSMISSION
Unknown.
This infected animal was one of a herd which lived in a large
swamp formed by the Luitikila river, about four miles from Mpika.
The sitatunga only emerge from the swamp in the late evening
and very early morning for an hour or two in order to feed, and
even then do not wander more than a few hundred yards from the
water.
Glossina morsitans have never been found within a radius of
fifteen miles, and it is therefore exceedingly improbable that they
are the vector.
Leeches are found in enormous numbers in the swamp, and feed
voraciously on human beings. It seems possible that these animals
are the vectors, more particularly as the parasite bears a very close
resemblance to amphibian trypanosomes.
Mosquitos are also extremely plentiful in this swamp. Filaria
were seen in the blood films of the same animal.
272
DESCRIPTION OF PLATE XXII
Trypanosoma tragelaphi. Films fixed in alcohol and stained
with Giemsa. The figures were drawn with the aid of the camera
lucida at a magnification of 2,000 diameters.
273
SECTION V
ON THE
DEVELOPMENT OF T. RHODESIENSE
IN GLOSSINA MORSITANS
BY
ALLAN KINGHORN, WARRINGTON YORKE
AND
LLEWELLYN LLOYD
In the course of our investigations, we endeavoured to
accumulate information regarding the development of the human
trypanosome in G. morsitans. Reference has already been made
to this subject in a previous paper,* and it is here intended to
correlate the facts at our disposal. It may be remarked at once
that owing to the comparatively small number of laboratory-bred
G. morsitans available, the information we have collected is by no
means so definite as could have been desired.
Up to the present, comparatively little work has been done on
this subject, and the records are more or less contradictory.
Kleine,t who was the first investigator to write on the development
of T. gambiense in G. pal pal is, is of the opinion that the complete
cycle takes place in, and is limited to, the intestine, whereas the
Royal Society Commissioners* in Uganda consider that involve¬
ment of the salivary glands is essential. They state that without
invasion of the salivary glands there is no infectivity of the fly.
TECHNIQUE
The method of dissection of the flies used by us was that
described by one of us in a previous paper. Briefly, it consists in
splitting the dorsum of the thorax longitudinally, and, after
separating the muscles and loosening the tissues with needles,
• Kinghom and Yorkc. Annals of Tropical Med. and Parasitology, 1912, Vol. VI.
t Kleine. 4 Trypanosome ns tudien.’ Arb. aus d. kaiserl. Gesundheitsamte, Bd. XXXI, Heft 2.
I Bruce, etc. Reports of the Roy. Soc. Commission in Uganda, 1911.
274
drawing out the salivary glands attached to the pharynx through
the waist. This method has obvious advantages over that described
by the Royal Society Commission, in which after snipping off the
terminal segment of the abdomen the whole contents were expressed
on to a glass slide, and the salivary glands subsequently separated
from the mass of intestines and other structures. We claim for the
technique adopted by us that the process is quicker, more certain,
and that the danger of contamination from the intestines is reduced
to a minimum. In fact, the only lesion in the alimentary canal
accompanying the operation occurs in the anterior portion of the
oesophagus.
To a certain point the information obtained from our
dissections is exceedingly definite. We found that in every fly
capable of infecting animals with the human trypanosome
( T. rhodesiense) the salivary glands were invaded. Of the 160
laboratory-bred Glossina morsitans utilised in various experiments
to transmit T. rhodesiense , 132 were dissected as they died. The
remaining twenty-eight were too dry when discovered to allow of
dissection. Twenty-seven of those dissected were found to be
infected with trypanosomes. The day of the experiment on which
the flies died and the results of dissection are given in Table 44.
A glance at the table shows that, of these 132 flies, five became
capable of infecting animals with the human trypanosome. In
each of these there was an enormous invasion of the salivary glands
by trypanosomes. In the 127 flies which remained incapable of
transmitting the parasite, the salivary glands were not involved,
although trypanosomes were found in the intestines of twenty-two.
A precisely comparable state of affairs was observed on
dissection of ‘ wild 1 Glossina morsitans which had become infective
after feeding on infected animals. The salivary glands were
found to be infected only in those insects which were capable of
transmitting the human trypanosome. In all, 906 ‘wild* Glossina
morsitans were used in these experiments, and of this number 620
were dissected. The remainder were for various reasons too dry to
admit of dissection. Of these 620 flies the salivary glands of
fourteen were invaded by trypanosomes. All except four of these
were definitely proved to transmit the human trypanosome. In the
case of the other four, the animal upon which the flies had been
275
Taili 44.—Results of dissection of laboratory-bred Gi. morsitans which were found to contain
parasites after being fed on infected animals.
Date of
infective
feed
Day of dissection
after infective
feed
Rjlsul
rs or Dissec
TION
Remarks
So.
Proboscis
Intestine
Salivary
gland
1
9.9.11
4
O
+ + +
O
2
10.8.12
4
O
+
O
3
22.1.12
5
O
+
O
4
9.8.12
5
O
+ +
O
5
10.8.12
5
O
+ + +
O
6
23.6.12
5
O
+
O
7
8.8.12
6
O
+ + +
0
8
9.8.12
6
O
+ + +
O
9
8.8.12
7
O
+ + +
O
10
23.6.12
7
O
+
O
11
10.8.12
8
O
+ +
O
12
8.8.12
9
O
+
O
*3
10.8.12
10
O
+ + +
O
*4
8.8.12
11
O
+ + +
O
'5
25.8.11
12
O
+ + +
O
16
31.8.11
12
O
+ + +
O
*7
8.8.12
>3
O
+ + +
O
18
5 - 3 >i 2
*5
+
+ + +
O
*9
11.8.12
*9
+ +
+ + +
+ + +
Infective on 12th day
20
10.8.12
20
+
+ + +
O
21
9.8.12
21
2 tryps.
seen
+ + +
O
22
9.8.12
21
O
+ + +
O
23
8.8.12
22
1 tryp.
seen
+ + +
O
24
8.8.12
22
+
+ + +
+ + +
Infective on 17th to
21 st day
25
v 4.11.11
28
O
+ + +
+ + +
On 15th
26
22.1.12
29
O
+ + +
+ + +
On 19th
27
9.9.11
40
O
+ + +
+ + +
On 13th
N.B.—O = Negative, + = Scanty, + + = Considerable numbers, -f -f + = Swarming.
276
allowed to feed died before a diagnosis could be made. None of
the 606 flies in which the salivary glands were not involved were
able to infect animals with the human trypanosome. Again, the
infectivity of Glosstna morsitans in nature was examined, as
mentioned in a previous section, by feeding batches of freshly-
caught flies on healthy monkeys. Certain of the groups infected
monkeys, and from one of these infective groups the actual infective
fly was isolated. This, on dissection, was found to have the
salivary glands swarming with trypanosomes. The remaining
242 flies in this group, which had been shown to be non-infective
when fed on monkeys, were dissected, and in no instance was an
infection of the salivary glands observed.
Table 45.— Results of dissection of wild Gl. morsitans found to be capable of infecting animals
with 7. rbodesiense .
No.
Date of
infective
feed
1
Day of
dissection after
infective feed
| Result of Dissection
Proboscis
Intestine
Salivary
glands
Remaiks
1
21.11.11
2 5
O
+ + +
4-4-4-
Infective on nth day.
2
21.11.11
2 5
O
+++
4-4-4-
Infective on nth day.
3
1.7.12
28
O
4-4-4-
4-4-4-
Infective on 13th day.
4
21.11.11
30
4-
+++
4-4-4-,
5
21.11.11
3 °
4-
4- 4* 4-
4-4-4-
Not proved to be infective.
6
21.11.11
30
O
4-4-4-
4-4-4-
7
21.11.11
30
+
4-4-4-
+++,
8
14.2.12
39
O
4-4- 4-
4-4-4-
Infective on 25th day.
9
4.10.11
40
O
4-4-4-
4-4-4-
Infective in nature.
10
30.6.12
4 2
O
4-4-4-
4-4-4-
Infective on 14th day.
11
30.6.12
47
O
4 + 4
4-4-4-
Infective on 14th day.
12
13.6.12
58
O
4- + +
+ 4--K
Infective on 48th day after
1
infected meal, or 8
*3
13.6.12
58
O
+ 4-4-
4-4-4-
days after having been
placed in the incubator.
14
13.6.12
59
O
4-4-4-
j
*5
14.6.12
7 1
O
4-4-4-
4-4-4-
Not proved to be infective,
but inoculation of try¬
panosomes from gut
and salivary glands
followed by positive
results.
V
277
In all, twenty Glossina morsitans were found to have invasion of
the salivary glands by trypanosomes, and of these, sixteen were
definitely found to be capable of infecting animals with
T. rhodesiense. Owing to unavoidable circumstances, we were
unable to prove the point in the case of the remaining four, but
there is nor reason to doubt that had the animals on which these flies
were fed survived beyond the necessary five or six days they would
have proved to be infected.
In order to anticipate the criticism that the trypanosomes were
not really inside the salivary glands, but lying outside these
structures, and due to contamination from the gut,- our examina¬
tions were conducted with extreme care. In the first place, the
glands were removed uninjured and attached to the pharynx,
placed on a microscope slide, and gently covered with a coverslip.
By careful focussing, it could easily be decided that the parasites
were actually in the lumen of the tubes and not outside. Moreover,
they were usually present in such enormous numbers as absolutely
to exclude the possibility that they were the result of contamination
from the intestine. Again, the glands of other flies were removed
with care and immediately fixed, and subsequently imbedded and
cut. In the sections the parasites could be seen to be inside the
glands. Finally, in order to remove any possibility of doubt,
sections of the whole abdomen of these infective flies were made,
and the glands found to be loaded with trypanosomes.
It will be seen from Table 44 that it was by no means a rare
occurrence for trypanosomes to be present in the intestines in the
earlier stages, especially in the case of those flies examined within
a few days of the infected meal. As a general rule, however, most
of the insects dissected after the first five or six days were
negative. In a certain proportion multiplication of the parasites
took place in the intestine.
As to the reason for this multiplication in the gut of occasional
flies only, and as to the manner in which it occurs, we have obtained
but little information. On one occasion a fly, which died on the
12th day after having been fed on a guinea-pig infected with
T. rhodesiense , was found to have an enormous gut infection.
Possibly there were also a very few trypanosomes in the salivary
glands, but on this point we could not be absolutely certain, as the
278
H
insect had been dead for some time before the dissection was made.
In the mid-gut were found a number of cysts containing swarms of
trypanosomes. Some of the cysts had thin walls and were filled
with a seething mass of flagellates, while others had thicker walls
and the contents were quiescent. The cysts ranged in diameter
from 27 to 32 p. Unfortunately, we are unable to state whether
the fly was infective at the time of death. It had refused to feed
for two or three days previously, and the animal on which it had
last fed (9th day of the experiment) did not become infected. The
gut contents were inoculated into a monkey, but the animal died
from some unknown cause a couple of days later.
Although multiplication of the parasites occurred in the guts of
a proportion of the flies, we met no instance in which a fly was
infective and in which inoculation of the gut parasites into experi¬
mental animals gave rise to infection, unless there was an
accompanying invasion of the salivary glands. On the other hand,
it appears that on every occasion on which the salivary glands are
involved the trypanosomes, both in these structures and also in the
intestines, are virulent, i.e., the fly infects when fed on a healthy
animal, and inoculation of the parasites from either the salivary
glands or the intestine gives rise to infection.
The results of inoculation of trypanosomes from laboratory-
bred flies in different stages of infection, and also from the wild
flies which were proved to be infective, are given in Table 46.
Our knowledge of the manner in which the salivary glands
become infected is uncertain, but there is a certain amount
of evidence which would cause one to believe that it is
secondary to the intestinal infection, and that it only occurs
when the trypanosomes in the gut have reached a certain
stage of development, and only then when the conditions
of temperature are suitable for the further development of the
parasites. In the first place, of 752 flies dissected at various
intervals after having fed on infected animals, we never found
trypanosomes in the salivary glands in the earlier stages before the
flies were infective. Again, whenever trypanosomes were found in
the salivary glands there were also enormous numbers present in the
intestine. Moreover, it is significant that whenever trypanosomes
were found in the salivary glands they were always infective, as
were also those present in the gut.
279
• Owing to the unfortunate death of the monkey on which thii fly was fed we were unable to ascertain whether the insect was infective or not. The fly was
one of the series which was kept for 60 days after the infective feed at laboratory temperature, and on the 6ist day placed in the incubator. In view of the fact
that the parasites in both the salivary glands and in the intestine were infective to subinoculated rats, it is highly probable that had the animal, on which the
fly was fed, lived long enough, it would have been found to be infected.
28o
Attention has already been drawn in a previous section to
experiments which suggest that although the parasites can multiply
and develop up to a certain stage in the intestine at comparatively
low temperatures (55°-65° F.), yet the flies do not become infective
until the temperature to which they are subjected is raised to at
least 75 °-8 o° F. In none of our experiments were trypanosomes
found in the salivary glands of flies which had not been subjected
to the higher temperatures. Probably the salivary glands become
invaded by parasites which have reached a certain stage in their
developmental cycle in the intestine. The remarkably short period
(eight days or less) in which three flies, which had been kept forty
days after the infective feed at laboratory temperature, became
infected after being placed in the incubator at 85° F. can be best
explained on the assumption that some portion of the cycle must
have occurred in the gut during the first forty days at laboratory
temperature.
Whilst the evidence that invasion of the salivary glands is
secondary to that of the intestine is fairly conclusive, our knowledge
of the path by which the parasites reach the glands from the
intestine and of the morphological characteristics of the forms
which migrate is by no means so definite.
It is at once apparent that there are two alternative routes by
which the trypanosomes might reach the salivary glands from the
gut, viz.: —(i) By way of the proboscis; (ii) by penetrating the
gut wall, and after crossing the coelom traversing the wall of
the salivary glands. We have accumulated no evidence enabling
us to decide along which of these two routes the parasites migrate.
Parasites were but rarely seen in the proboscis, and then, with one
exception, only in small numbers. In the case referred to, trypano¬
somes were present in the proboscis in considerable numbers, but as
both the gut and salivary glands contained enormous numbers of
flagellates, their occurrence in the proboscis could easily be
explained on the assumption that they had been discharged from
the salivary glands and were passing down the proboscis with the
secretion of these structures. The parasites from the gut, from the
salivary glands, and from the proboscis were inoculated into
monkeys and rats, all of which became infected, showing that the
trypanosomes in each of these structures were virulent.
281
As will be seen from Plate XXIII, the parasites present in the
gut and in the salivary glands of infective flies exhibit marked
differences in morphological characteristics. The forms encountered
in the intestine were many and diverse, but the predominant type
was undoubtedly that depicted in figs. 7-12. It may be described as
a large broad flagellate, with a feebly-developed undulating
membrane and little or no free flagellum. The nucleus is usually
compact and situated near the centre of the body, but not
infrequently it lies in a more posterior position. The blepharoplast
is, as a rule, situated near the posterior extremity, but sometimes
lies further forward approaching the nucleus. In addition to these
forms, long thin parasites were found. Here the nucleus was, as
a rule, diffuse, and occupied a central position. The parasites
stained more faintly, the undulating membrane was narrow, and
there was generally a distinct free flagellum. Many more or less
rounded bodies were also seen.
The form met with in the salivary glands of infective flies was
almost invariably that represented by figs. 1-5. It approximates
somewhat closely to the short form of the trypanosome in the blood
of the vertebrate host, but is obviously not identical with this. The
nucleus is compact, and is situated at the middle of the body; the
blepharoplast is distinct, and lies near the posterior extremity.
The undulating membrane is well developed; there is only
occasionally a short free flagellum. The length of this form is
15-18/4. In addition to this form, long attenuated flagellates were
occasionally seen, but they were very few in number, and it was
necessary to search for a considerable period before discovering one
of them. These forms resemble the corresponding variety seen in
the gut. The nucleus is diffuse, and situated near the centre of the
body. The parasite stains a faint pink with Giemsa.
In so far as we could determine, the predominant gut type was
not met with in the salivary glands, and the predominant salivary
gland type did not occur in the intestine. In spite of this,
however, we are faced with the fact that the parasites in both
situations proved to be virulent when inoculated into experimental
animals. It appears to us that two hypotheses can be advanced to
explain this; either that the salivary type is the only virulent form
and that a certain number of these reach the intestine from the
284
DESCRIPTION OF PLATE XXIII
Trypanosoma rhodesiense in Glossina morsitans. Films f
in alcohol and stained with Giemsa. The figures were drawn •
the aid of a camera lucida at a magnification of 2,000 diameter
Figs. 1-5. Typical forms seen in the salivary glands.
Figs. 6-14. Intestinal forms.
X
A-J.BrQufieijj.dei
TRyp *SOSO»i
N C-LOSVm
'-"ESESSE
•GIST,
ORY
continuously
May, 1912.
id, and has
native fowls
re used. At
ards entirely
*d feedings,
e flies were
ble food, in
/ upon them
nachs of the
iis condition
n found to
d as blood
Dody cannot
>ort. The
is not been
inders, five
three flies,
was found
: damaging
die through
1 deposits,
of blotting
en be taken
placing the
5 are kept
es becomes
re kept.
276
allowed to feed died before a diagnosis could be made. None of
the 606 flies in which the salivary glands were not involved were
able to infect animals with the human trypanosome. Again, the
infectivity of Glossina morsitans in nature was examined, as
mentioned in a previous section, by feeding batches of freshly-
caught flies on healthy monkeys. Certain of the groups infected
monkeys, and from one of these infective groups the actual infective
fly was isolated. This, on dissection, was found to have the
salivary glands swarming with trypanosomes. The remaining
242 flies in this group, which had been shown to be non-infedive
when fed on monkeys, were dissected, and in no instance was an
infection of the salivary glands observed.
Table 45.—Results of dissection of wild Gl. morsitans found to be capable of infecting animal*
with T. rbodestrnse.
I Result of Dissection
Date of
Day of
No.
infective
dissection after
Salivary
Remaiks
feed
infective feed
Proboscis
Intestine
glands
1
21.11.11
*5
O
++ +
+ 4* +
Infective on 11 th day.
2
21.11.11
2 5
O
+ + +
+ + +
Infective on nth day.
3
1.7.12
28
O
+ + +
+ + +
Infective on 13th day.
4
21.11.11
30
+
+ + +
+ + + ,
5
21.11.11
30
+
+ + +
+ + +
V
Not proved to be infective.
6
21.11.11
3 °
O
+ + +
+ + +
7
21.11.11
3 °
+
+ + +
+ + + ,
8
14.2.12
39
O
+ + +
+++
Infective on 25th day.
9
4.10.11
40
O
+ + +
+ + +
Infective in nature.
10
30.6.12
4 2
O
+ + +
+ + +
Infective on 14th day.
11
30.6.12
47
O
+ + +
4 - + +
Infective on 14th day.
12
13.6.12
58
O
+ ++
+ 4- + v
Infective on 48th day after
infected meal, or S
13
13.6.12
58
O
+ + +
4-4-4-
days after having been
placed in the incubator.
*4
13.6.12
59
O
+ + +
4-4-4-)
1 S
14.6.12
7 1
O
+ -f +
4-4- +
Not proved to be infective,
but inoculation of try¬
panosomes from gut
and salivary gland*
followed by positive
results.
277
In all, twenty Glossina morsitans were found to have invasion of
the salivary glands by trypanosomes, and of these, sixteen were
definitely found to be capable of infecting animals with
T. rhodesiense. Owing to unavoidable circumstances, we were
unable to prove the point in the case of the remaining four, but
there is ncr reason to doubt that had the animals on which these flies
were fed survived beyond the necessary five or six days they would
have proved to be infected.
In order to anticipate the criticism that the trypanosomes were
not really inside the salivary glands, but lying outside these
structures, and due to contamination from the gut,- our examina¬
tions were conducted with extreme care. In the first place, the
glands were removed uninjured and attached to the pharynx,
placed on a microscope slide, and gently covered with a coverslip.
By careful focussing, it could easily be decided that the parasites
were actually in the lumen of the tubes and not outside. Moreover,
they were usually present in such enormous numbers as absolutely
to exclude the possibility that they were the result of contamination
from the intestine. Again, the glands of other flies were removed
with care and immediately fixed, and subsequently imbedded and
cut. In the sections the parasites could be seen to be inside the
glands. Finally, in order to remove any possibility of doubt,
sections of the whole abdomen of these infective flies were made,
and the glands found to be loaded with trypanosomes.
It will be seen from Table 44 that it was by no means a rare
occurrence for trypanosomes to be present in the intestines in the
earlier stages, especially in the case of those flies examined within
a few days of the infected meal. As a general rule, however, most
of the insects dissected after the first five or six days were
negative. In a certain proportion multiplication of the parasites
took place in the intestine.
As to the reason for this multiplication in the gut of occasional
flies only, and as to the manner in which it occurs, we have obtained
but little information. On one occasion a fly, which died cm the
12th day after having been fed on a guinea-pig infected with
T. rhodesiense , was found to have an enormous gut infection.
Possibly there were also a very few trypanosomes in the salivary
glands, but on this point we could not be absolutely certain, as the
278
insect had been dead for some time before the dissection was made.
In the mid-gut were found a number of cysts containing swarms of
trypanosomes. Some of the cysts had thin walls and were filled
with a seething mass of flagellates, while others had thicker walls
and the contents were quiescent. The cysts ranged in diameter
from 27 to 32 fi. Unfortunately, we are unable to state whether
the fly was infective at the time of death. It had refused to feed
for two or three days previously, and the animal on which it had
last fed (9th day of the experiment) did not become infected. The
gut contents were inoculated into a monkey, but the animal died
from some unknown cause a couple of days later.
Although multiplication of the parasites occurred in the guts of
a proportion of the flies, we met no instance in which a fly was
infective and in which inoculation of the gut parasites into experi¬
mental animals gave rise to infection, unless there was an
accompanying invasion of the salivary glands. On the other hand,
it appears that on every occasion on which the salivary glands are
involved the trypanosomes, both in these structures and also in the
intestines, are virulent, i.e., the fly infects when fed on a healthy
animal, and inoculation of the parasites from either the salivary
glands or the intestine gives rise to infection.
The results of inoculation of trypanosomes from laboratory-
bred flies in different stages of infection, and also from the wild
flies which were proved to be infective, are given in Table 46.
Our knowledge of the manner in which the salivary glands
become infected is uncertain, but there is a certain amount
of evidence which would cause one to believe that it is
secondary to the intestinal infection, and that it only occurs
when the trypanosomes in the gut have reached a certain
stage of development, and only then when the conditions
of temperature are suitable for the further development of the
parasites. In the first place, of 752 flies dissected at various
intervals after having fed on infected animals, we never found
trypanosomes in the salivary glands in the earlier stages before the
flies were infective. Again, whenever trypanosomes were found in
the salivary glands there were also enormous numbers present in the
intestine. Moreover, it is significant that whenever trypanosomes
were found in the salivary glands they were always infective, as
were also those present in the gut.
272
DESCRIPTION OF PLATE XXII
Trypanosoma tragelafhi. Films fixed in alcohol and stained
with Giemsa. The figures were drawn with the aid of the camera
lucida at a magnification of 2,000 diameters.
273
SECTION V
ON THE
DEVELOPMENT OF T. RHODESIENSE
IN GLOSSINA MORSITANS
BY
ALLAN KINGHORN, WARRINGTON YORKE
AND
LLEWELLYN LLOYD
In the course of our investigations, we endeavoured to
accumulate information regarding the development of the human
trypanosome in G. morsitans. Reference has already been made
to this subject in a previous paper,* and it is here intended to
correlate the facts at our disposal. It may be remarked at once
that owing to the comparatively small number of laboratory-bred
G. morsitans available, the information we have collected is by no
means so definite as could have been desired.
Up to the present, comparatively little work has been done on
this subject, and the records are more or less contradictory.
Kleine,t who was the first investigator to write on the development
of T. gambiense in G. falpalts, is of the opinion that the complete
cycle takes place in, and is limited to, the intestine, whereas the
Royal Society Commissioners* in Uganda consider that involve¬
ment of the salivary glands is essential. They state that without
invasion of the salivary glands there is no infectivity of the fly.
TECHNIQUE
The method of dissection of the flies used by us was that
described by one of us in a previous paper. Briefly, it consists in
splitting the dorsum of the thorax longitudinally, and, after
separating the muscles and loosening the tissues with needles,
* Kinghom and Yorke. Annals of Tropical Med. and Parasitology, 1912, Vol. VI.
t Kleine. 4 Trypanosomenstudien.* Arb. aus d. kaiserl. Gesundheitsamte, Bd. XXXI, Heft 2.
X Bruce, etc. Reports of the Roy. Soc. Commission in Uganda, 1911.
2 74
drawing out the salivary glands attached to the pharynx through
the waist. This method has obvious advantages over that described
by the Royal Society Commission, in which after snipping off the
terminal segment of the abdomen the whole contents were expressed
on to a glass slide, and the salivary glands subsequently separated
from the mass of intestines and other structures. We claim for the
technique adopted by us that the process is quicker, more certain,
and that the danger of contamination from the intestines is reduced
to a minimum. In fact, the only lesion in the alimentary canal
accompanying the operation occurs in the anterior portion of the
oesophagus.
To a certain point the information obtained from our
dissections is exceedingly definite. We found that in every fly
capable of infecting animals with the human trypanosome
( T. rhodesiense ) the salivary glands were invaded. Of the 160
laboratory-bred Glossina morsitans utilised in various experiments
to transmit T. rhodesiense , 132 were dissected as they died. The
remaining twenty-eight were too dry when discovered to allow of
dissection. Twenty-seven of those dissected were found to be
infected with trypanosomes. The day of the experiment on which
the flies died and the results of dissection are given in Table 44.
A glance at the table shows that, of these 132 flies, five became
capable of infecting animals with the human trypanosome. In
each of these there was an enormous invasion of the salivary glands
by trypanosomes. In the 127 flies which remained incapable of
transmitting the parasite, the salivary glands were not involved,
although trypanosomes were found in the intestines of twenty-two.
A precisely comparable state of affairs was observed on
dissection of 1 wild * Glossina morsitans which had become infective
after feeding on infected animals. The salivary glands were
found to be infected only in those insects which were capable of
transmitting the human trypanosome. In all, 906 'wild’ Glossina
morsitans were used in these experiments, and of this number 620
were dissected. The remainder were for various reasons too dry to
admit of dissection. Of these 620 flies the salivary glands of
fourteen were invaded by trypanosomes. All except four of these
were definitely proved to transmit the human trypanosome. In the
case of the other four, the animal upon which the flies had been
275
Table 44.— Results of dissection of laboratory-bred Gl. morsitans which were found to contain
parasites after being fed on infected animals.
1
| Date of
infective
feed
Day of dissection
after infective
feed
Results or Dissection
Remarks
No.
Proboscis
Intestine
Salivary
gland
1
9.9.11
4
O
+ + +
O
2
10.8.12
4
O
+
O
3
22.1.12
5
O
+
O
4
9.8.12
5
O
+ +
O
5
10.8.12
5
O
+ + +
O
6
23.6.12
5
O
+
O
7
8.8.12
6
O
+ + +
O
8
9.8.12
6
O
+ + +
O
9
8.8.12
7
O
+ + +
O
10
23.6.12
7
O
+
O
11
10.8.12
8
O
+ +
O
12
8.8.12
9
O
+
O
13
10.8.12
10
O
+ + +
O
*4
8.8.12
11
O
+ + +
O
15
25.8.11
12
O
+ + +
O
16
31.8.11
12
O
+ + +
O
17
8.8.12
13
O
+ + +
O
18
5 - 3 ii*
>5
+
+ + +
O
*9
11.8.12
>9
+ +
+ + +
+ + +
Infective on 12th day
20
10.8.12
20
+
+ + +
O
21
9.8.12
21
2 tryps.
seen
+ + +
O
22
9.8.12
21
O
+ + +
O
23
8.8.12
22
1 tryp.
seen
+ + +
O
24
8.8.12
22
+
+ + +
+ + +
Infective on 17th to
21 st day
25
14.11.11
28
O
+ + +
+ + +
On 15th
26
22.1.12
29
O
+ + +
+ + +
On 19th
27
9.9.11
40
O
+ + +
+ + +
On 13th
N.B.—O = Negative, + « Scanty, + +
Considerable numbers, + -f + = Swarming.
276
allowed to feed died before a diagnosis could be made. None of
the 606 flies in which the salivary glands were not involved were
able to infect animals with the human trypanosome. Again, the
infectivity of Glossina morsitans in nature was examined, as
mentioned in a previous section, by feeding batches of freshly-
caught flies on healthy monkeys. Certain of the groups infected
monkeys, and from one of these infective groups the actual infective
fly was isolated. This, on dissection, was found to have the
salivary glands swarming with trypanosomes. The remaining
242 flies in this group, which had been shown to be non-infective
when fed on monkeys, were dissected, and in no instance was an
infection of the salivary glands observed.
Table 45.—Result* of dissection of wild Gl. morsitans found to be capable of infecting animals
with T. rhodesiense.
No.
Date of
infective
feed
Day of
dissection after
infective feed
Result of Dissection
Remarks
Proboscis
Intestine
Salivary
glands
1
21.11.11
2 5
O
+ + +
+ + +
Infective on nth day.
2
21.11.11
2 5
O
+++
+ + +
Infective on nth day.
3
1.7.12
28
O
+++
+ + +
Infective on 13th day.
4
21.11.11
30
+
+ + +
+ + +
5
21.11.11
30
+
+ + +
+ + +
y
Not proved to be infective.
6
21.11.11
3 °
O
+ + +
+ + +
7
21.11.11
3 o
4 -
+ + +
+ + + j
8
14.2.12
39
O
+ + +
+ + +
Infective on 25th day.
9
4.10.11
40
O
+ + +
+ + +
Infective in nature.
10
30.6.12
4 2
O
+ + +
+ + +
Infective on 14th day.
ii
30.6.12
47
O
+ + +
+ + +
Infective on 14th day.
12
13.6.12
58
O
+ + +
+ 4- + *
Infective on 48th day after
infected meal, or S
13
13.6.12
58
O
+ + +
4- 4- 4-
days after having been
placed in the incubator.
*4
13.6.12
59
O
+ + +
+ + + J
>5
14.6.12
7 *
O
+ + +
4-4- +
Not proved to be infective,
but inoculation of try¬
panosomes from gut
and salivary glands
followed by positive
results.
2 77
In all, twenty Glossina morsitans were found to have invasion of
the salivary glands by trypanosomes, and of these, sixteen were
definitely found to be capable of infecting animals with
T. rhodesiense. Owing to unavoidable circumstances, we were
unable to prove the point in the case of the remaining four, but
there is nor reason to doubt that had the animals on which these flies
were fed survived beyond the necessary five or six days they would
have proved to be infected.
In order to anticipate the criticism that the trypanosomes were
not really inside the salivary glands, but lying outside these
structures, and due to contamination from the gut,- our examina¬
tions were conducted with extreme care. In the first place, the
glands were removed uninjured and attached to the pharynx,
placed on a microscope slide, and gently covered with a coverslip.
By careful focussing, it could easily be decided that the parasites
were actually in the lumen of the tubes and not outside. Moreover,
they were usually present in such enormous numbers as absolutely
to exclude the possibility that they were the result of contamination
from the intestine. Again, the glands of other flies were removed
with care and immediately fixed, and subsequently imbedded and
cut. In the sections the parasites could be seen to be inside the
glands. Finally, in order to remove any possibility of doubt,
sections of the whole abdomen of these infective flies were made,
and the glands found to be loaded with trypanosomes.
It will be seen from Table 44 that it was by no means a rare
occurrence for trypanosomes to be present in the intestines in the
earlier stages, especially in the case of those flies examined within
a few days of the infected meal. As a general rule, however, most
of the insects dissected after the first five or six days were
negative. In a certain proportion multiplication of the parasites
took place in the intestine.
As to the reason for this multiplication in the gut of occasional
flies only, and as to the manner in which it occurs, we have obtained
but little information. On one occasion a fly, which died cm the
12th day after having been fed on a guinea-pig infected with
T. rhodesiense , was found to have an enormous gut infection.
Possibly there were also a very few trypanosomes in the salivary
glands, but on this point we could not be absolutely certain, as the
2 7 8
insect had been dead for some time before the dissection was made.
In the mid-gut were found a number of cysts containing swarms of
trypanosomes. Some of the cysts had thin walls and were filled
with a seething mass of flagellates, while others had thicker walls
and the contents were quiescent. The cysts ranged in diameter
from 27 to 32 fi. Unfortunately, we are unable to state whether
the fly was infective at the time of death. It had refused to feed
for two or three days previously, and the animal on which it had
last fed (9th day of the experiment) did not become infected. The
gut contents were inoculated into a monkey, but the animal died
from some unknown cause a couple of days later.
Although multiplication of the parasites occurred in the guts of
a proportion of the flies, we met no instance in which a fly was
infective and in which inoculation of the gut parasites into experi¬
mental animals gave rise to infection, unless there was an
accompanying invasion of the salivary glands. On the other hand,
it appears that on every occasion on which the salivary glands are
involved the trypanosomes, both in these structures and also in the
intestines, are virulent, i.e., the fly infects when fed on a healthy
animal, and inoculation of the parasites from either the salivary
glands or the intestine gives rise to infection.
The results of inoculation of trypanosomes from laboratory-
bred flies in different stages of infection, and also from the wild
flies which were proved to be infective, are given in Table 46.
Our knowledge of the manner in which the salivary glands
become infected is uncertain, but there is a certain amount
of evidence which would cause one to believe that it is
secondary to the intestinal infection, and that it only occurs
when the trypanosomes in the gut have reached a certain
stage of development, and only then when the conditions
of temperature are suitable for the further development of the
parasites. In the first place, of 752 flies dissected at various
intervals after having fed on infected animals, we never found
trypanosomes in the salivary glands in the earlier stages before the
flies were infective. Again, whenever trypanosomes were found in
the salivary glands there were also enormous numbers present in the
intestine. Moreover, it is significant that whenever trypanosomes
were found in the salivary glands they were always infective, as
were also those present in the gut.
279
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280
Attention has already been drawn in a previous section to
experiments which suggest that although the parasites can multiply
and develop up to a certain stage in the intestine at comparatively
low temperatures (55°-65° F.), yet the flies do not become infective
until the temperature to which they are subjected is raised to at
least 75 °-8 o° F. In none of our experiments were trypanosomes
found in the salivary glands of flies which had not been subjected
to the higher temperatures. Probably the salivary glands become
invaded by parasites which have reached a certain stage in their
developmental cycle in the intestine. The remarkably short period
(eight days or less) in which three flies, which had been kept forty
days after the infective feed at laboratory temperature, became
infected after being placed in the incubator at 85° F. can be best
explained on the assumption that some portion of the cycle must
have occurred in the gut during the first forty days at laboratory
temperature.
Whilst the evidence that invasion of the salivary glands is
secondary to that of the intestine is fairly conclusive, our knowledge
of the path by which the parasites reach the glands from the
intestine and of the morphological characteristics of the forms
which migrate is by no means so definite.
It is at once apparent that there are two alternative routes by
which the trypanosomes might reach the salivary glands from the
gut, viz.:—(i) By way of the proboscis; (ii) by penetrating the
gut wall, and after crossing the coelom traversing the wall of
the salivary glands. We have accumulated no evidence enabling
us to decide along which of these two routes the parasites migrate.
Parasites were but rarely seen in the proboscis, and then, with one
exception, only in small numbers. In the case referred to, trypano¬
somes were present in the proboscis in considerable numbers, but as
both the gut and salivary glands contained enormous numbers of
flagellates, their occurrence in the proboscis could easily be
explained on the assumption that they had been discharged from
the salivary glands and were passing down the proboscis with the
secretion of these structures. The parasites from the gut, from the
salivary glands, and from the proboscis were inoculated into
monkeys and rats, all of which became infected, showing that the
trypanosomes in each of these structures were virulent.
28i
As will be seen from Plate XXIII, the parasites present in the
gut and in the salivary glands of infective flies exhibit marked
differences in morphological characteristics. The forms encountered
in the intestine were many and diverse, but the predominant type
was undoubtedly that depicted in figs. 7-12. It may be described as
a large broad flagellate, with a feebly-developed undulating
membrane and little or no free flagellum. The nucleus is usually
compact and situated near the centre of the body, but not
infrequently it lies in a more posterior position. The blepharoplast
is, as a rule, situated near the posterior extremity, but sometimes
lies further forward approaching the nucleus. In addition to these
forms, long thin parasites were found. Here the nucleus was, as
a rule, diffuse, and occupied a central position. The parasites
stained more faintly, the undulating membrane was narrow, and
there was generally a distinct free flagellum. Many more or less
rounded bodies were also seen.
The form met with in the salivary glands of infective flies was
almost invariably that represented by figs. 1-5. It approximates
somewhat closely to the short form of the trypanosome in the blood
of the vertebrate host, but is obviously not identical with this. The
nucleus is compact, and is situated at the middle of the body; the
blepharoplast is distinct, and lies near the posterior extremity.
The undulating membrane is well developed; there is only
occasionally a short free flagellum. The length of this form is
15-18/1. In addition to this form, long attenuated flagellates were
occasionally seen, but they were very few in number, and it was
necessary to search for a considerable period before discovering one
of them. These forms resemble the corresponding variety seen in
the gut. The nucleus is diffuse, and situated near the centre of the
body. The parasite stains a faint pink with Giemsa.
In so far as we could determine, the predominant gut type was
not met with in the salivary glands, and the predominant salivary
gland type did not occur in the intestine. In spite of this,
however, we are faced with the fact that the parasites in both
situations proved to be virulent when inoculated into experimental
animals. It appears to us that two hypotheses can be advanced to
explain this; either that the salivary type is the only virulent form
and that a certain number of these reach the intestine from the
282
salivary glands by way of the proboscis—this may be quite
fortuitous, depending on the fact that a certain quantity of infected
saliva is from time to time drawn into the intestine—and are then
lost amongst the multitude of diverse gut forms, or that there is
some unknown form (the one which migrates from the intestine to
the salivary glands) which precedes the typical salivary gland type
and which is also virulent for laboratory animals.
In conclusion, we might remark that invasion of the salivary
glands was only observed in the case of flies infected with the
human trypanosome (T. rhodesiense ) and not in the case of any of
the other trypanosomes with which we had to deal either in the
Luangwa Valley or on the Congo-Zambesi watershed. This was
the case both with the strain of T. rhodesiense derived from man
and also with that found in ' wild * Glossina morsitans which had
been infected in nature.
It is of interest to note that of 310 ‘wild’ Glossina morsitans
which were dissected as they were brought into the laboratory
recognisable mammalian red corpuscles were found in the intestine
of seventy, whilst nucleated red corpuscles were only found on
twelve occasions.
SUMMARY
1. The salivary glands of all Glossina morsitans capable of
transmitting T. rhodesiense are infected, and conversely without
invasion of the salivary glands there is no infectivity of the fly.
2. Invasion of the salivary glands is secondary to that of the
intestine.
3. The first portion of the developmental cycle of the trypano¬
some takes place in the gut. In order for its completion and for
invasion of the salivary glands to occur, a relatively high mean
temperature, 75°-85° F., is necessary.
4. Invasion of the salivary glands was only found in flies
infected with the human trypanosome, T. rhodesiense.
5. The predominant type of the trypanosome in the intestine
of infected Glossina morsitans —a large broad form—is quite
283
different from that which predominates in the salivary glands,
where the parasite resembles somewhat the short form seen in the
blood of the vertebrate host.
6. Both the intestinal forms and also those from the salivary
glands of infective Glossina morsitans are virulent when inoculated
into healthy animals.
284
DESCRIPTION OF PLATE XXIII
Trypanosoma rhodesiense in Glossina morsitans. Films fixed
in alcohol and stained with Giemsa. The figures were drawn with
the aid of a camera lucida at a magnification of 2,000 diameters.
Figs. 1-5. Typical forms seen in the salivary glands.
Figs. 6-14. Intestinal forms.
. l////<//s Tro/i Mtu/. (< /hntsifoL, I of \U
Plate XXLl /
A. M. B rookfieId,deI
TRYPANOSOMA RH ODESIEN SE
IN GLOSSINA MORSITANS
285
SECTION VI
REPORT OF THE ENTOMOLOGIST,
LLEWELLYN LLOYD
(a) CLOSSINA MORSiTANS IN THE LABORATORY
The breeding of these flies was carried out continuously
from the middle of June, 1911, to the end of May, 1912.
Kinghom* has already outlined the method adopted, and has
described the larva and pupa. The flies were fed on native fowls
at Nawalia, except for a short period when goats were used. At
Ngoa the flies were fed at first on fowls, but afterwards entirely
upon goats, which died quickly under the repeated feedings,
doubtless owing to the trypanosomes with which the flies were
naturally infected. Fowls would appear to be unsuitable food, in
spite of the fact that the flies feed much more readily upon them
than upon mammals. Fowl blood in the ‘sucking’ stomachs of the
flies forms large firm clots, and apparently blood in this condition
cannot be utilised by the flies, as the clots have been found to
persist for several weeks, monkeys having been used as blood
donors in the interim. A fly with so large a clot in the body cannot
retain a full-grown larva, and consequently many abort. The
phenomenon is rare with mammalian blood, and it has not been
observed in nature.
The bottles used for the stock flies were glass cylinders, five
inches deep and two inches in diameter. At first only three flies,
two females and a male, were kept in each tube, but it was found
later that the number could be increased to six without damaging
the vitality of the insects. In damp weather many flies die through
fouling themselves with the copious semi-fluid faecal deposits.
This may be obviated by inverting the tubes over sheets of blotting
paper, which absorb the excreta, but precautions must then be taken
to avoid the escape of the larvae. This was done by placing the
tubes in dishes and paper-lined boxes. When flies are kept
under these conditions the daily changing of the tubes becomes
unnecessary—a great saving of labour when many flies are kept.
Kinghom, A. Bulletin Entomological Research, 1912.
286
It is sometimes difficult to obtain a sufficient supply of female
flies, but natives learn to distinguish between the sexes and to
select and bring in females. In a collection made without
discrimination, the proportion of females is often as low as 2 %.
During the twelve months the number of females in stock has varied
considerably. From July to February, with the exception of a
period at the end of August when all were destroyed by an
accident, the number varied between 200 and 300. During April
and May the number of females was between 500 and 600. The
flies breed most readily at the beginning and at the end of the rains.
This is confirmed by observations on the number of flies in nature
at different periods of the year.
Males and females emerged from the pupae in approximately
equal proportions for the twelve months. From June to December
the males in each month were somewhat in excess of the females,
while from January to April the reverse was the case. The
pupation period of a male is on the average about a day longer
than that of a female.
The variation in the duration of the pupation period is given in
Table 47, p. 288. It shows more fully the influence of the
climatic conditions on the pupal life; the pupae have been divided
into groups which were deposited in periods of half months. For
each of these periods the mean temperature in the laboratory and
the relative humidity of the air are given. A fuller account of. the
meteorological conditions is given in a previous section of this report.
The approximate mean temperatures to which the various groups of
pupae were exposed have been calculated by averaging the mean
temperatures for the periods in which the groups were deposited, and
for those periods following which could have influenced the
duration of the pupation period. Chart 8 indicates how
readily the pupae respond to changes of temperature. The
humidity of the air has little or no influence on the pupal life, as
will be seen by comparing the data of June and July with those of
the second half of March. While the relative humidity of the
latter portion was 10 % higher than in the former, the temperature
and the duration of the pupal life were approximately the same in
each case. It should be mentioned that the observations from June
to the middle of March were made at Nawalia (altitude 2,100 feet),
Cmamt 8.— Showing the Influence of temperature on the pupation period of Glostina morsttans.
7 y n l V V 3 J. A/V3W
288
Table 47.—Showing the influence of Temperature on the Pupation Period of Glouina morn tans.
Period during
which the pupae
were obtained
Number of
apparently
healthy pupae
Number of
pupae from which
flies emerged
Average duration
of pupation
period—days
Mean laboratory
temperature
Approximate
mean temperature
to which pupae
were exposed
Relative
humidity
of air
June 26—30
3
3
5 1
—
67° F.
48-6%
July 1 —15 .
6
5
47
64-!°F.
70
—
O'*
1
2 4
18
39
68-3
1
71
457
August 1—15 ..
21
9
35
694
73
—
„ 16—31 ..
5
1
33
73 *o
7 6
35*8
September 1—15 ..
2
2
2 5
7 2 7
82
—
„ 16—JO ..
*4
12
2 4
807
83
3**5
October 1—15 .
22
*5
*3
82*9
85
—
„ 16—31 ..
20
IO
2 3
86-4
8S
31-8
November 1—15 ..
l 7
6
2 4
86-o
83
—
» 16—30 ...
2 5
16
26
8**5
80
41* 1
December 1—15 ..
2 7
2 3
2 7
80 *6
79
—
„ 16—31 ..
*3
12
28
78-8
78
69*1
January 1—15 ..
20
.6
28
79 -°
78
—
„ 16—31 ..
3 *
28
2 9
P
00
r>»
77
777
February 1—15 ..
48
3 2
3 1 !
77 * 2
77
; —
„ 16—29 ..
40
30
1 3 o
76-9
77
73*8
March 1—2
11
8
3 °
00
6
77 !
!
—
„ 21—26 ..
4
3
48
75*9
69 !
625
April 1—15 ••
2 5
16
54
77 * 2
67
—
f
0
5 1
22
7 °
68-0
64
537
May 1—15 ••
7 i
20*
81
70*0
62
—
i» 16 — 3 1
96
—
63:9
—
5*-3
June 1—15 ••
—
—
61-8
—
—
,, 16—30 ..
t
—
—
—
56-9
—
53 *o
• On August 4 the remaining pupae were placed in an incubator.
289
while those for the remainder of the time were made at Ngoa
(altitude 4,400 feet).
The shortest pupation period was that of a pupa deposited in
the middle of October, from which the fly emerged in twenty-one
days. The mean temperature to which this pupa was exposed was
slightly over 86° F. The longest period was eighty-eight days for
a pupa obtained during May and exposed to a mean temperature of
62° F. Reference to the table will show that these extremely high
and low temperatures have a very deleterious effect on the pupae,
evinced by the high percentage from which flies do not emerge.
Many of the flies which emerged after periods of over seventy days
were malformed or weak, and did not survive well. The mean
shade temperature of the outside air was always higher than that
of the laboratory, and the majority of the pupae found in nature
in July were placed in such positions that for several hours a day
they would be exposed to the sun, except for a slight covering of
soil. It is therefore probable that these very long pupation periods
do not occur under natural conditions.
(6) A RECORD OF SOME BREEDING PLACES OF
GLOSSINA MORSITANS
The pupae of Glossina morsttans were found in a variety of
positions at Ngoa in July; all in the neighbourhood of two
permanent streams, the Kanchibia and the Kalamba. At the time,
the middle of the dry season, the leaves of the deciduous trees were
falling rapidly, and dense shade was only to be found actually in
the beds of the streams. The bush is of the nature of an open wood
with a large variety of trees. The big trees grow, in general, right
to the banks of the streams without intervening plains. Tsetse fly
are very numerous in this area.
The following is a list of the positions in which pupae have
been found: —
(1) Under a thick branch (native name Mutobo) running about
half an inch above the surface of the soil, on gently sloping ground
fifty yards from a stream, one living and one empty pupa case
were found. They were close to the base of the branch, and were
lightly covered with soil.
290
(2) A similar position under a branch of a tree (native name
Muombo) twenty yards from the stream, five cases and numerous
fragments were found among dead leaves and humus.
(3) In a hollow in the trunk of a tree (native name Mpasa) a few
inches above the surface of the ground; ten yards from the stream.
The bottom of the hollow measured about four inches by two, and
was full of a firm clay impregnated with resinous matter. Ten
pupae, nine cases and numerous fragments were taken from the
crevices in the surface, several of them quite exposed.
This spot, visited a few days later, yielded another pupa. The
sun shines into this hollow for several hours a day, striking the
surface of the soil.
(4) In two large hollows (filled with an old termite nest much
broken down) at the base of a tree of the same species on the bank
of the stream. One of the hollows was well sheltered and received
no sun. In this, fifty cases and numerous fragments were found,
but no living pupae. The other hollow received the afternoon sun,
and in it one living pupa and several cases were found. (PI. XXIV,
fig. 1).
(5) In a well-sheltered hollow between three main branches of
a tree (native name Mupapa), containing dead leaves but no soil
or humus, one pupa and eight cases occurred. This tree was on the
bank of the stream, and the hollow was about two feet above the
ground. (PI. XXIV, fig. 2.)
(6) In a shallow cup full of soil and roots in a tree (native name
Mutobo) close to a stream. This hollow was quite exposed to the
sun. One empty case was taken.
(7) One pupa and two cases were found in a hollow full of
insect droppings, two feet from the ground, in a tree (native name
Muombo). The hollow was exposed to the sun, and the tree was
two hundred yards from the stream.
(8) This position resembles the first two. One pupa and one
case were found under the trunk of a tree (native name Muanga)
which ran for six feet close to the ground before rising. The tree
is four hundred yards from water. (PI. XXV, fig. 3.)
(9) Three empty cases were found in two shallow cups, quite
exposed to sun and rain, in a Mutobo tree. The cups contained a
little dust and were four feet from the ground, being formed by a
swelling on the trunk caused by the pollarding to which the
Annals Trop. Med. & Parasitol .. Yol. VII
PLATE XXIV
Fig. i. Pupal habitats of Glossitia morsitans
Annals Trop. Med. & ParasitolVol. I'll
PLATE XXV
Fig. 3. Pupal habitats of Glossina morsitans.
Fig. 4. Pupal habitats of Glossina morsitans .
To face p. 291
C. Tin ling Co., Ltd., Imp.
291
Awemba tribes subject the trees when making gardens. Three cases
were found.
(10) A shallow in a Muombo tree, three feet from the ground,
full of loose soil and roots and quite exposed. The tree was
two hundred yards from water. Three pupae and five cases were
taken.
{it) The stump of a rotten tree built around by termites, which
still occupied the base. Twelve living pupae and many fragments
were found under the branch and in and about the termite nest.
The position was not shaded, and the stump would be well warmed
by the sun.
(12) A main branch of a Mutondo tree had been cut across, the
cut surface facing upwards. The wood had rotted away internally,
forming a cup full of humus and exposed to the sun. Ten living
pupae and four cases were found here. (PI. XXV, fig. 4.)
(13) Two pupae and two cases were found in a hollow in an
Mpasa tree two hundred yards from water. The hole was eighteen
inches above the ground, and was full of insect droppings.
(14) In the deserted burrow of an ant-bear (Orycteropus
capensis). The hollow had been occupied and enlarged by warthog
during the last rainy season. The burrow ran in such a manner
that the morning sun would shine in obliquely or to one side. On
this side, and near the opening, nine living pupae and six empty
cases were found, but no pupae were found on the sheltered side. In
a similar burrow near, which was sheltered from the sun, no pupae
were found. This locality was about three hundred yards from
water. (PI. XXVI, fig. 5.)
(15) In a burrow of an ant-bear half a mile from water one
empty case was found.
(16) Two living pupae were found in a ‘salt-lick’ in the side
of an ant-hill two hundred yards from water. The position was
exposed to the sun. These * salt-licks ’ are very common in the
district, and are visited by most of the big mammals which scrape
and lick away the soil for the sake of the salt it contains until large
excavations are formed.
(17) A hollow one foot from the ground in a Musangati tree,
full of hard clay and well sheltered, contained two cases.
(18) Ten cases were taken under the trunk of a Mpasa tree
292
running close to the ground. The position was well sheltered, and
was twenty yards away from the water.
(19) A hollow between two main branches of a Chimpampa tree
contained two cases. The hollow was filled with dead leaves and
humus, and was well sheltered. The tree was close to water.
The pupae were thus found in association with trees of eight
different species and in holes in the earth. The native names only
of the trees can be given at present. All the trees about which they
were found were either abnormal or injured. None have been
found at the bases of normal trees or under bushes. The pupae
were generally, though not always, hidden under slight covering of
earth or dead leaves. These observations confirm those of Jack*,
who first recorded the finding of the pupae in nature, that they are
deposited in such positions that they are not likely to be scratched
up by game birds.
The relation of the positions to shade is of interest, and is
probably connected with the season. Of the fifty-four living pupae
found, forty-nine (90 %) were so placed that they would
be daily warmed by the sun. They were collected in the coldest
part of the year, and at a time when flies were emerging from pupae
in the laboratory after a period of from seventy-five to eighty-five
days. The shade temperatures outside were at the time about
2° F. higher than those of the laboratory, so that in well-shaded
positions the pupation period would be about seventy days. This
protracted period would be much reduced in pupae which were
exposed to the warming influence of the sun.
Reference to Table 47 giving the relation of the pupation period
to the mean temperature will show that for temperatures below
70° F., an increase of i° in the mean temperature causes a reduction
in the pupation period of from three to five days. Several of these
sunny positions would be waterlogged during rain, and therefore
unsuitable for the deposition of pupae.
•Jack, R. W. Bull, of £nt. Res., Vol. 11 , pp. 357-361.
Austen, £. £. A Handbook of the Tsetse Flies, London, 191 x, p. 56.
Anr.ah Trop. Med. & ParasttolVol. VI1
PLATE XXVI
Fig. 5. Pupal habitats of Glossina morsitans.
To face f. jgj
C. Tinting d-* Co., Ltd., Imp.
293
(c) A RECORD OF BLOOD-SUCKING INSECTS AND TICKS
COLLECTED IN THE LUANGWA VALLEY FROM AUGUST, 1911,
TO MARCH, 1912, AND AT NGOA FROM MARCH TO MAY, 1912
lxodoidea
Ornithodorus moubata , the Nkufu tick, was taken in three
villages in the neighbourhood of Chinunda. In two instances it
occurred in single huts in the villages in very large numbers, but was
said not to have been seen in any other huts. In the third village
it was found in two huts, in one of which it was numerous, while in
the other, a deserted hut, a single specimen was taken. In this
village the information was given that the pest had occurred in
almost every hut, but had disappeared from all but the one. The
absence of this pest from the Nawalia district is peculiar, as it must
be constantly introduced from the higher ground where it is common.
The natives attribute the absence to the heat, but a number of
Nkufu brought to Nawalia from the above-mentioned villages lived
and bred in the laboratory throughout the hot weather.
Tabanidae
These insects, which are very numerous both as regards species
and individuals in the Luangwa Valley, were very troublesome in
the laboratory, to which they were doubtless attracted by the
numbers of animals about the place. Seventeen species of the
genus Tabanus alone were taken in the laboratory at different times.
Chrysops. Only one species, C. fuscipennis, Ric., was seen, and
a single specimen was taken at Nawalia in February. It flew out
of long grass over swampy ground and settled on the hand. It was
so eager to feed that it was readily caught in a tube.
Tabanus
Insects of this genus appeared spasmodically from August to
October, the driest months of the year. In November, after the first
showers of rain, they become much more numerous, and continued
to appear in large numbers throughout the wet months, becoming
scarcer again in March. In the latter month several species
reappeared which had not been seen for some time, indicating that
they were probably emerging from the pupae about this time.
Males of several species were also taken in March. The following
species were collected: —
T. africanus , Gray. This species was taken at intervals at
Nawalia during the rainy season. It was most common
during February and March, when it was frequently seen
in the laboratory.
T. bigut tat us, Wied. Only three specimens of this large
species were seen. They were taken in the laboratory and
in a canoe in November and February. The males,
usually described as being commoner than the females,
were not seen.
T. par., Walk. This was one of the most numerous species
around Nawalia. It constantly entered the laboratory,
and was’ most persistent in its biting. First seen in
October, it became numerous in January, and its numbers
declined towards the end of the rains.
T. liventipes, Sure. This somewhat rare species was taken in
small numbers at Nawalia in January and February.
Specimens were taken on kudu, on roan and about the
camp.
T . nigrostriatus, Ric. Hitherto recorded only from Nyasaland
and in small numbers, this species was one of the
commonest and most troublesome at Nawalia. It entered
the laboratory in large numbers, and was often seen on
dead game during November and December. It gives very
painful bites, settling especially on the head and biting
through the hair.
T. taeniola , P. de B. This fly was taken first in October, and
became common in November and December at Nawalia.
It was again taken in some numbers in March. During the
latter month a male was taken at Ngoa.
T. taeniola, var. variatus , Wlk. This variety was much more
numerous than the typical species. It was first taken in
September biting a native at sunset. It was not seen again
till November, when it became very troublesome. During
January and February it was not observed, but was again
taken in some numbers in March.
T. fraternus , Macq. Never very common; specimens of this
species were taken at Nawalia in August, November,
December and February.
2 95
T. fuscipes, Ric. This species first appeared in January at
Nawalia, and continued to be numerous to the middle of
March. Both sexes were seen in large numbers resting on
long grass over a swamp in the evening about the middle
of January. In February and March about ninety female
specimens of this fly were dissected in connection with the
work of the Commission. The gradual maturing of the
eggs was thus observed, and early in the latter month some
had deposited part of their ova.
T. albipalpus , Wlk. A single female specimen of this species
was taken at Ngoa in March. The species has hitherto
been regarded as belonging to the West African Fauna,
all the specimens having been taken on the West Coast
with the exception of one which is recorded from Uganda.
T. pullulus , Acert. This species was taken at Nawalia in small
numbers from January to March.
T. claritibialis , Ric. First taken in the laboratory at Nawalia
in January. This species was fairly numerous in the two
following months.
T. atrimanus , Lw. A single female of this fly was taken in the
laboratory at Nawalia during October.
T. copemani , Aust. This species, which has been previously
recorded from the Feiera district of Northern Rhodesia
and from Nyasaland, was first taken at Nawalia about the
middle of January, and in the two following months was
one of the commoner species.
T . diversus , Ric. A female specimen of this species was taken
in a tent at Kasongo on the Luangwa river in October.
One other specimen was found dead on a table in the
laboratory, at Nawalia in December. It had been caught
by a spider, and was densely wrapped up in a web.
T. maculatissinus , Macq. One female of this was taken
feeding on a rhinoceros at Ngoa in May. The distance
from water was about a quarter of a mile, and the insect
flew to the animal shortly after it had been shot.
In addition to the above, five other species of Tabanus were
collected at Nawalia and Ngoa. One of these, I am informed, is
an undescribed species, specimens of which have previously reached
296
the British Museum from various localities, and which will shortly
be described. The remaining species have not yet been identified.
Haematopota
This genus was not nearly so conspicuous as the preceding one,
either as regards species or individuals. One specimen was taken
in December, and they were more numerous in January and
February. After the latter month they were extremely scarce.
The following species have been identified : —
H. mactans, Aust. A single specimen was taken on a kudu in
January at Nawalia.
H. insidiatrix, Aust. This species was several times taken in
the laboratory at Nawalia during January and February,
and it was also collected on antelope.
H. sp. An undescribed species which has previously been
collected in Northern Rhodesia. Several were taken on
antelope.
Muscidae. Glossina. The only species of this genus that has been
collected is Glossina morsitans, Westwood. In the district to which
the work of the Commission has been confined this insect is generally
distributed. While small areas exist, such as the immediate
neighbourhood of Mpika, in which the fly is not actually resident,
there is probably no spot in which it does not occasionally occur.
Auchmeromyia luteola, F., is common both in the Luangwa
Valley and on the Plateau. During September a large number of
huts were searched, and one or more stages of this insect were seen
in every hut. The adult insect is to be found throughout the year.
Cordylobia anthropophaga , Grunb. The larone of this fly were
several times seen in the legs of wild rats. One larone which was
squeezed out of its sac burrowed in sand and pupated, emerging as
a fly after a very brief pupation period.
Slomoxys nigra, Macq. A few specimens of this species were
collected at Nawalia in January. They were found resting on
leaves and grass over a swampy stream.
Stomoxys calcitrans, Linn. (?) was very numerous at Mpika in
March about the cattle kraals and on some dogs. In May, when
some were required for experimental purposes, only one or two
297
individuals were found. At this time there had been no rain for
two months.
Pupipara
Hippobosca hirsuta , Aust. This species was taken repeatedly
on waterbuck ( Cobus ellipsiprymnus and C. defassa), and on puku
( C . vardoni ), both in the Luangwa Valley and at Ngoa. On one
occasion a specimen was taken on the back of a native who had been
present when a waterbuck was shot a few hours earlier.
Two other species of the Hippoboscidae were collected at
Nawalia. One of these, a wingless species, was taken on several
occasions on bushbuck ( Tragelaphus scriptus ), while the other, a
winged species, was only taken on one occasion on hartebeest
(Bubalis lichtensteini ).
My thanks are due to the Entomological Research Committee
for the identification of many of the above insects. It
would be difficult to exaggerate the usefulness of this institution to
Entomologists in the field.
APPENDIX A
AN EXPERIMENT TO ASCERTAIN
WHETHER TABANIDS TRANSMIT
TRYPANOSOMES IN NATURE
BY
A. F. WALLACE, M.B.
M.O., N. KHODUIA.
AND
LLEWELLYN LLOYD
An experiment to determine whether flies of the genus Tabanus
were vectors of trypanosomes in nature was carried out at Nawalia.
As pressure of work prevented this being done at the time when
Tabanids were most numerous, the number of flies used was small
and the results are inconclusive. The experiment was carried out
during February and March, the last two months of the wet season.
Each morning a number of fly boys were sent out to collect the
flies, and brought in daily from ten to thirty, the number being
larger when the sun was shining. These were placed in separate
tubes, and were fed on a monkey. Only a small proportion of the
insects placed on the animal took food. It seemed to make no
difference whether the feeding was done in direct sunlight or in the
shade.
All the flies that fed were dissected and examined the same day.
The dissection of a Tabanus is a very simple matter. A needle is
thrust through the head and the thorax is held with forceps. The
fly is then immersed in salt solution, and a series of gentle pulls is
given to the head. The chitin of the neck breaks across, and the
alimentary canal is drawn out with the salivary glands. The
gut breaks across in the weakest place, just in front of the
proventriculus, and the head comes away with the salivary glands
attached. To obtain the remainder of the gut in an unbroken
condition, the last segment of the abdomen is cut round with
3 °o
needles, and the gut then drawn out backwards. The proventriculus
passes back readily through the thorax and brings with it the
trilobed ‘ sucking stomach.’ Microscope preparations were then
made of the proboscis with the pharynx, the complete gut from
proventriculus to anus, and the salivary glands. These preparations
were examined for parasites.
In all, 128 flies were fed and dissected. These comprised the
following species: —
T. fuscipes, Ric.
T. taeniola, P. de B
T. claritibialis, Ric.
T. copematti, Aust.
T. pullulus, Aust.
T. par, Wlk. ...
87 flies.
25 „
2 „
10 „
2 „
2 ,,
These flies were fed between February 7th and March 15th.
The monkey did not become infected.
Flagellated parasites were found in seven flies. In no cases
were they seen in the proboscis or the salivary glands, but were
confined to the mid- and hind-gut. Smears were made of the
preparations, and in four instances inoculations were made into wild
rats. Only two of the rats lived long enough for any conclusions
to be drawn, and these remained negative.
3 oi
APPENDIX B
AN ATTEMPT TO TRANSMIT
TRYPANOSOMA RHODESIENSE
BY MEANS OF ORNITHODOROS
MOUBATA
BY
A. F. WALLACE
An attempt was made at Nawalia to transmit T. rhodesiense by
Ornithodoros moubata. For this purpose ticks had to be brought
down to Nawalia from the plateau at Mpika. It is a curious fact
that there are no ‘ nkufu ’ (the native name for these ticks) in the
Luangwa Valley, but that they abound in the hilly country on either
side. This fact is well recognised by the natives, who explain it by
the Luangwa Valley being too hot for ticks to live in. This may
be the explanation of the high death rate in the experimental ticks.
Five experiments were conducted, about twenty-five ticks being
used in each. Each group of ticks was fed on a monkey heavily
infected with T. rhodesiense , and after an interval of about a
month was fed on a clean monkey. Each lot of ticks was kept
under observation for six to seven months, being fed at intervals on
clean monkeys. Only a few of the ticks survived for seven months.
The following is a summary of the experiments: —
To begin with, 125 ticks were fed on a heavily-infected monkey.
After one month 87 ticks were fed on a clean monkey.
,, two months 51 ,,
„ three „ 8 „
„ six „ 23 „
„ seven „ 10 ,,
The blood of the clean animals remained negative.
99 99 99 99
9 9 9 9 99 99
99 99 99 99
99 99 99 99
302
The result of interrupted feeding was also investigated. After
the ticks had become partially distended with blood they were
removed from the infected monkey and transferred to healthy
animals. Certain of the ticks would not continue their meal, and
in the case of those which did there was always a short interval,
varying from a half to five minutes before they resumed. In all,
sixty-one ticks were used in these experiments to ascertain whether
the trypanosome could be transmitted from one infected animal to
a healthy one by means of interrupted feeding. The result was
negative in every instance.
3°3
ON THE NON-IDENTITY OF TRYPANO¬
SOMA BRUCEI{ Plimmer and Bradford, i 899)
WITH THE TRYPANOSOME OF THE
SAME NAME FROM THE UGANDA OX'
BY
J. W. W. STEPHENS, M.D., D.P.H. (Cantab.)
AND
B. BLACKLOCK, M.D., D.P.H.
(.Received for publication 7 December , 1912)
INTRODUCTION
Before considering our own observations it will be necessary to
review briefly previous statements regarding the morphology of
Trypanosoma brucei.
(1) Bruce 1 states that the haematozoa vary among themselves a
good deal in shape and size and seem to take on slightly different
forms in different species of animals. He publishes four figures
depicting nine trypanosomes. Possibly one or two of the five
figured from the dog might be considered to be 1 stumpy * forms.
(2) Kanthack, Durham, and Blandford 2 state that the Nagana
parasites vary considerably both in size and form. They may be
long and pointed and sometimes stouter, some individuals are
short and thick with a short flagellum, their protoplasm being
crowded with granules. This description suggests dimorphism,
but it should be noted that forms without a free flagellum are not
mentioned. No slides were available belonging to these observers,
but Dr. Durham kindly lent us a large series of photographs. On
examining these, one or, perhaps, two show a * stumpy ’ form, but it
is difficult to be certain, and the uniformity of the remainder is
striking. They state that 'the material for our observation was
obtained in the first instance from the blood of a dog infected by
* Read before the Royal Society of London on January 23, 1913, and reprinted from Proc.
Roy. Soc., B, Vol. LXXXVI, pp. 187—191.
3®4
the disease on the voyage from Africa, and brought to England in
November, 1896, by Dr. Waghom.’
This animal we believe to be the origin of the strain of T. brucei,
Plimmer and Bradford, 1899, described by these authors, and at
present maintained in England, so far as we can gather, solely at
Liverpool.
It is not stated above from what animal the dog was infected
on the voyage, nor is it stated what the exact original source of the
strain derived from Zululand was.
(3) Plimmer and Bradford 3 ’ 4 describe four forms in the blood,
but neither their description nor figures suggest that they have seen
stumpy forms. They describe ‘ a large hyaline form.’ * This
organism is much larger than the ordinary adult form, and is much
wider, often more than double the width, and is more irregular in
shape. The protoplasm is quite homogeneous and much more
delicate, and it stains very faintly with the methylene blue.’ They
are still to be found in films, and we easily found them in
Dr. Plimmer’s old films, but we found only extremely rarely forms
that could be called ‘stumpy.’ We think that if they had been
present these observers would hardly have failed to have noticed
and drawn attention to them, as they have a striking appearance.
(4) Bruce and others 5 make a comparison between T. brucei,
Uganda, 1909, and T. brucei, Zululand, 1894.
They state that many of the old Zululand preparations are still
extant, so that it has been possible to do this. The preparations
were, however, 15 years old, and had been stained with carbol
fuchsin. The slides were got from horse, donkey, ox, monkey,
dog; 200 trypanosomes were measured from the Zululand strain
and 172 from the Uganda strain. The curves obtained in this way
certainly resemble one another, though in one case the peak is at
i8/«, in the other case at 20 fi.
Also trypanosomes are figured from each strain, and there can
be little doubt that there is a close resemblance, if not identity,
viz., in the fact that both possess both long and stumpy forms.
‘ With the evidence available the Commission consider themselves
justified in considering the trypanosome recovered from the Uganda
ox to be identical with T. brucei, the cause of Nagana in Zululand
and other parts of South Africa.’
3°5
Further, Bruoe states in another paper 8 that T. brucei (Uganda
strain) has actually 26 per cent, of non-flagellated forms.
(5) In 1911 Laveran 7 published an article, entitled ‘ Identification
et essai de classification des trypanosomes des mammif&res.’
In this article he places T. brucei in his Group I, ‘ Trypanosomes
chez lesquels le flagelle pr&ente toujours une partie libre,’ whereas
he places T. gambiense in his Group III, ‘ Trypanosomes ayant des
formes a flagelle libre et des formes sans flagelle libre.’
Or, in other words, T. brucei is classed among the monomorphic
trypanosomes while T. gambiense is among the dimorphic. We
have then two opposite statements as to the morphology of
T. brucei, (1) that it is monomorphic and (2) that it is dimorphic.
We possess two strains of (so-called) T. brucei in the laboratory,
viz., the Zululand strain, of which we have given the origin above,
and the Uganda strain from Surgeon-General Bruce, obtained
originally from the ox in Uganda in 1909. These strains have
been maintained continuously at Runcorn in a variety of animals,
the Zululand strain for 4^ years, and the Uganda strain for
2\ years. In the case of the Uganda strain it was lost in 1912 for
a short period but was returned to us again by Prof. Mesnil, who
had previously received it from us.
We made then a preliminary examination of these two strains,
and found to our surprise that they could easily be distinguished
morphologically.
We next proceeded to make a detailed examination of the two
strains in a series of slides throughout the entire period of infection
in various animals, viz., rats, guinea-pigs, and rabbits. As the
result, we believe we have established the following facts: —
(a) The Zululand strain is typically monomorphic. The
trypanosomes are long, with a long free flagellum. We must
admit, however, that it is possible (as we believe is the case also in
another typically monomorphic trypanosome, viz., T. evansi), to
find by long search short forms which somewhat resemble true
stumpy forms, but we must emphasise the fact that in all the slides
we have examined, prolonged search is necessary to find them.
( b ) We have also verified the fact that Laveran’s T. brucei
strain also is, as he says, monomorphic. The origin of this strain
seems uncertain. Laveran probably received it from Ehrlich, but
306
where the latter got it from cannot now be ascertained. Unless it
came from England, there must be two monomorphic T. brucei
strains in existence, not to mention the possibility of other T, brucei
strains of uncertain parentage in various laboratories.
We have examined also old slides from the Zululand strain
lent us by Prof. Nuttall, Colonel Skinner, R.A.M.C., and
Dr. Plimmer.* All these were monomorphic. We repeat here that
in these films, or at least some of them, it was possible by long
search to find a short form somewhat resembling a stumpy form,
but not having the somewhat indefinable characteristic appearance
of the latter.
(r) The Uganda strain, on the contrary, is typically dimorphic y
i.e., besides the usual long forms of trypanosomes, stumpy forms
are readily found, even in abundance occasionally, when the
infection is well marked. Bruce, 6 as we have noted above, states
that this trypanosome has 26 per cent, of non-flagellated forms.
The typical stumpy form we may define as a short, thick
trypanosome, 12-14*1, almost straight or slightly curved along one
edge, while along the other the membrane is thrown into bold folds,
there being no free flagellum, or at times a very short or doubtfully
free one.
It is thus easy to distinguish a typical Uganda specimen from a
Zululand specimen, and, in fact, we may express the difference in
this way, that it is impossible to match a typical Uganda slide by
any slide from the Zululand strain.
We have stated in the above history of the Uganda strain that
it was recently returned to us by Prof. Mesnil, who remarked in his
letter that he had maintained it in mice (for nearly a year), and
that it showed now very few 4 trapues 1 forms. This we have been
able to verify in the films made from the infected mouse sent to us.
But, as soon as we had re-inoculated it into guinea-pigs, it again
showed numerous stumpy forms. But the same does not hold good
for the Zululand strain; in guinea-pigs, as in rats and rabbits, the
strain is typically monomorphic , i.e., it does not show stumpy
forms. We therefore conclude that the two strains, as we now
possess them in the laboratory, are different.
* We desire here to express our thanks to these gentlemen for their kindness in sending
us slides.
30 ?
How, then, are we to explain these facts P There seem to us
three possibilities: —
1. That the strain we now possess, which we have been
designating T. brucei, Zululand, is not this strain at all, but some
other trypanosome inoculated erroneously during the course of
inoculations extending over years. We think this view is untenable,
for it would not explain the monomorphic character of the old
slides we have examined, nor would it explain Laveran’s mono¬
morphic trypanosome.
2. While Bruce may have been working with a dimorphic
trypanosome in Zululand, and still has slides showing these
characters, it is quite possible that the strain sent by him to England
was something quite different. This is all the more likely, as Bruce
successfully infected dogs from a variety of wild game, viz.,
wildebeeste, kudu, bush buck and buffalo, and, as Bruce himself
states, ‘when T. brucei was discovered in Zululand in 1894, it was
naturally thought to be the one and only trypanosome in Africa/
and no suspicion arose at that time of a multiplicity of trypano¬
somes in native game.
This is the simplest explanation, and the fact that Plimmer and
Bradford do not describe or figure stumpy forms, and our examina¬
tion of Dr. Plimmer’s slides had the same result, makes it probable
that this is the true one.
3. That the strain originally sent to England was dimorphic,
but that it has now become monomorphic. This may have come
about in two ways : —
(a) The strain originally was a mixture of a long trypanosome
and a stumpy trypanosome, and the stumpy has now died out. If
this explanation were valid, it would probably imply that
T. gambiense and other dimorphic trypanosomes were also mixtures.
This we regard as a not impossible view, but one we cannot at
present prove or disprove.
( b ) The strain was originally dimorphic (but not a mixture), and
that it has now become monomorphic . If this were so, it would
modify materially our notions of specificity of trypanosomes, at
least in laboratories. Of such a change we have at present not much
evidence. We have noted, however, above that the Uganda strain
308
kept in mice for a year was almost (but not entirely) monomorphic,
but that in guinea-pigs it at once showed its normal characters.
It is impossible at present to decide between these explanations.
We come back, therefore, to the fact of which we ourselves have
no doubt, viz., that the trypanosome that Plimmer and Bradford
worked with, and which they named T. brucei in 1899, is certainly
now a monomorphic trypanosome, and is not the same as the
trypanosome from the ox described under the same name by Bruce
and others in Uganda.
We believe, then, that the facts we have brought forward prove
the non-identity of the Zululand and Uganda strains.
In order to avoid confusion, we think it advisable that this
Uganda trypanosome should be re-named. We therefore propose
for it the name T. ugandae.
REFERENCES
1. Bruce. * Further Report on the Tsetse-fly Disease or Nagana in Zululand/ by Surgeon-
Major David Bruce, A.M.S. (1896 ?). Harrison & Sons, London.
2. Kanthack, Durham St Blandford. Roy. Soc. Proc., 1898, Vol. LXIV, p. 100.
3. Plimmer St Bradford. ‘ A Preliminary Note on the Morphology and Distribution of the
Organism Found in the Tsetse-fly Disease,' Roy. Soc. Proc., 1899-1900, Vol. LXV,
p. 274.
4. Bradford, J. R., St Plimmer, H. G. * The Trypanosoma brucei , the Organism Found in
Nagana or Tsetse-fly Disease,' Quart. Joum. Micro. Sci., 1902, Vol. XLV, p. 449.
5. Bruce & Others. Reports of the Sleeping Sickness Commission of the Royal Society, 1911,
No. XI, p. 147.
6. Bruce. Ibid., 1912, No. XII, p. 24.
7. Laver an. ‘ Identification et essai de classification des trypanosomes des mammifires/
Ann. de l'lnstitut Pasteur, July, 1911, No. 7, p. 497.
3°9
CONCERNING THE SEX AND AGE
OF AFRICANS SUFFERING FROM
TRYPANOSOMIASIS
BY
JOHN L. TODD
(Received for publication 26 March , 1913)
An analysis (Table II) of the 79 cases of trypanosomiasis seen
among 12,298 natives of the Gambia Protectorate 1 shows that
76% of those found to be infected are adults, 24% of them are
children and none are aged; there are more boys (16*4%) than girls
(7*6%) among the cases, but the proportion of men and women
infected is approximately equal. It is probable that these figures
approximate a usual incidence of the disease in the Gambia since all
of the cases seen there in 1902 occurred in children and middle-aged
adults. 2
An analysis (Table I) of the results obtained in 1911 by the
examination of 9,069 Gambian natives, whose posterior cervical
triangles were palpated for enlarged glands, shows that a slight
degree of glandular enlargement is common among children of both
sexes, but especially in boys—probably because boys are less careful
of their person than are girls. Much enlarged * + 9 glands occur
most often in young adults—76% of the cases of trypanosomiasis
seen in the Gambia in 1911 belong to this class—and trypanosomes
were found in every person, with much enlarged glands, who was
examined for them.
Kinghom 4 noted that all of 89 cases of trypanosomiasis seen by
him on the Gold Coast were under 42 years of age; most of them
were young adults. He also reported 5 that glandular enlargement
is much more common in males than in females, and in children
than in adults.
In Nyasaland, Sanderson 8 reported that of 47 cases of
trypanosomiasis 383% were women and 53*2% were men,
3io
Number of individuals of 9,069 2,024 2,200 490 i,411 2,283
each class examined
8 5% were boys, while no girls were infected. Hear9ey 7 also points
out that, in Nyasaland, two cases are found among men for every
one seen in a woman.
The observations mentioned in these reports have much in
common. They all record the occurrence of more cases of trypano¬
somiasis among adults than among natives of any other class.
Unfortunately, the ages of the natives examined and the degree of
their glandular enlargements are not all recorded in the same way,
so that it is not possible to compare these observations exactly with
those made in the Gambia. But, because there is a general
similarity between all of these observations, it seemed advisable to
examine the records of natives seen in the Congo in 1903, 3 in order
to ascertain whether the same conditions had existed there.
Almost 90% of the total number of cases of trypanosomiasis
seen in the Congo Free State were in young adults (Table II). It
was thought that these figures might give a wrong idea of the
relative incidence of the disease at different ages, because, for two
reasons, most of the natives examined were young adults; first,
because natives of the Congo were unaccustomed to Europeans, and
the tendency was for women, children and the aged to hide from
the members of the expedition; and, second, because, by the
opening up of the country, large numbers of young men, sometimes
with their women, were drawn to the European settlements to act as
labourers and soldiers—such persons were easily examined, and the
cases of trypanosomiasis seen in European posts came almost
entirely from among them. Consequently, the cases of trypano¬
somiasis seen in the Congo are divided (Table II) into two groups
according as they occur in populations subjected to these selective
influences or not. The total number of cases dealt with is
comparatively small, but it is interesting, nevertheless, that the
total percentage of young adults infected is the same in each group.
Since the selected population contains many more males than
females, it is only natural that the percentage of infected males is
greater among cases of trypanosomiasis coming from it than among
those drawn from a population not subjected to selective influences.
With the exception that the percentage of young people among
the cases is greater, the incidence of trypanosomiasis in the Gambia
differs slightly from its incidence in the Congo. The difference can
312
3*3
be accounted for, in great part at least, by the much larger
proportion of adolescents among the Gambian natives examined;
compare the figures, in Tables I and III, stating the percentages of
the total number of natives examined, represented in each locality,
by children, adults and old persons. In the Gambia, where the
natives are habituated to the presence of Europeans, the huts were
entered and, often, every member of a village was examined; but,
even there, it is probable that many timid girls were not seen, while
the large percentage of boys recorded is the result of their
willingness to be examined. In order that it may be possible to
compare the observations made in the Congo with those made in
the Gambia, the figures for the Congo given in Table III, as far as
possible, record only examinations made in native villages where no
evident avoidance of the members of the expedition was manifest,
and where the nature of the population was uninfluenced by
economic conditions.
The results obtained by gland palpation in the Congo
(Table III) agree with' those obtained in the Gambia (Table I) in
the general absence of enlarged glands in old persons; they are
dissimilar in the larger percentages of children and of adults with
considerable degrees (‘ + 9 and ‘ H-’) of glandular enlargement.
There is certainly a close connection between these increased
percentages of enlarged glands and the much more frequent
occurrence of trypanosomiasis among natives of the Congo; it has
already been made clear that, in the Congo 3 as well as in the
Gambia, 1 enlarged cervical glands mean trypanosomiasis. In 1911,
it was estimated that 0*8% of the native population in the Gambia
had trypanosomiasis; there, cases only occur sporadically. 1 It is
scarcely possible to estimate the probable percentage infected among
the natives examined in the Congo in 1903-05, but it is certain that
30%, and often more, of the natives in many of the villages visited
were infected 8 ; nothing approaching such an epidemic exists in the
Gambia.
The smallness of the numbers recorded for old persons in all
of the tables is striking. The figures given for the Gambia
(Table I) are probably not very incorrect; though they doubtless
tend to err, in the same way as those given for the Congo
(Table III) which, for reasons already given, are probably lower
3i5
than they should be for children, especially girls, and for the
aged. There is no means of determining whether the numbers of
individuals of each class examined approximate those normally
existing in an African population, because vital statistics have not
been compiled for that continent. But, it seems probable, from an
inspection of statistics for European races, that fewer old persons
and more boys were seen than might have been expected. In 1900,
males over 45 constituted 9^%- of the population of Massachusetts,
and there were 11 % of females over 45; a cursory inspection of the
Scotch and Irish census returns for 1911 suggests that these figures
are, if anything, rather below those normal for a European
population. The proportion of children, especially boys, recorded
for the Gambia (Table III) is considerably larger than that in
Massachusetts, where about 11% of the population were boys under
13 and 10% were girls under 11 in 1900; although as large a
percentage of girls are recorded by the census of 1900 in parts of
Scotland as were seen in the Gambia the percentage of boys there
was about four points less than it was in the Gambia.
Table IV has been drawn up in order to ascertain how the
percentages of natives, belonging to groups limited by sex and age
characters, would vary in natives of the Congo living in districts
infected or uninfected by trypanosomiasis. The natives recorded
in it are taken from those whose glands were palpated in
populations unsubjected to the selective influences mentioned above
(Table II). Old persons are again strikingly absent among
natives from both infected and uninfected districts. As might be
expected, the proportion of adults, especially of men, is much less
among natives of infected districts. The death rate among them
from trypanosomiasis is greatest, because they are most exposed to
infection by their occupations. 1 It is well-known and many
observers have reported that fishermen and riverine tribes are
especially liable to be infected by trypanosomiasis. 8 p - 27
In appraising the value of the figures stated in this paper, it is
necessary to consider both the methods employed in collecting the
data on which the figures are based and the form in which they
are presented. All of the natives considered in Tables I and III
were apparently healthy persons, and the clinically recognisable
cases of sleeping sickness, seen among these populations, are not
3i6
3*7
included in these tables. The age of the natives examined was
roughly estimated by their appearance, and the degree of their
glandular enlargement was ascertained by a single, rapidly-made
examination; it is obvious that slight errors might easily occur both
in the estimate of age and the statement of the degree of glandular
enlargement; as has been described already, a proportion of the
population examined certainly escaped observation. The age
periods used in classifying the natives contain an unequal number
of years and the periods used for males and females are not the
same; these unusual age limits were chosen because they represent, 1
approximately, the periods during which male and female natives
follow the occupations of children, adults and elderly people.
Therefore, in comparing the figures given in these tables, it must be
remembered that errors in compilation have occurred and that the
span of years, allotted by the classification employed, is for adults
almost thrice that apportioned to children; consequently, a meaning
can be attached only to very considerable and constant differences
in the figures stated for each class. Finally, the total number of
infected and uninfected natives observed is too small; but in spite
of this, and although the methods employed are not sufficiently
accurate to permit far-reaching conclusions, it does seem as though
something may be learned from a cautious consideration of the
figures given in the tables accompanying this paper. It is believed
that a part of the large percentage of adults with enlarged glands
and a part of the large percentage of adults among cases of
trypanosomiasis may be due to the greater number of years
apportioned to the adult class, but it is also believed that most of
both percentages is due to an increased incidence of trypanosomiasis
among adults. That adults should be most infected is reasonable
since their occupations—washing, drawing water, fishing, farming
and travelling—expose them most to infection.
Trypanosomiasis, more prevalent in the Congo than in the
Gambia, is doubtless one of the reasons why the proportion of old
people is less among natives of the Congo than among those of the
Gambia, and much less among both populations than might be
expected. Even although a large proportion of adults infected
with trypanosomiasis and untreated die within three or four years 9
after contracting their infection, and although the aged are not
3 i8
exposed to infection by their occupation, it is astonishing that the
percentage of old persons among the cases of sleeping sickness and
the percentage of old persons with much enlarged glands should be
so small; it seems impossible that even so small a number of persons
could live during their years among populations so much infected
with trypanosomiasis without having been infected with the disease
themselves. The only explanation which suggests itself is that
some natives may be immune to trypanosomiasis. 1 * 2 * 5 * 10 If an
immunity does exist there are many reasons and analogies to
indicate that it is not an absolute, ‘ sterilizing * immunity but a
relative one in which the host is tolerant of the infecting parasite.
It is very much hoped that the natives, found to be infected in the
Gambia in 1911, will be kept track of, as can be done easily by the
Commissioners of the districts in which they were seen; a record of
the course of the disease in them would add considerably to
our knowledge of the duration and outcome of infection by
Trypanosoma gambiense in Africans.
The conclusions concerning the natives of the Congo and of the
Gambia, which can be drawn from the facts presented and discussed
in this paper, are stated in the following paragraphs.
1. The proportion of elderly individuals among them is lower
than it is among Europeans (Tables I, III, and IV).
2. By far, the majority of cases of trypanosomiasis are persons
of middle age; almost none of them are elderly persons (Table II).
3. The percentage of individuals with a considerable degree of
glandular enlargement—which is coincident with trypanosomiasis—
is very much greater in adults, and in children, than in elderly
persons (Tables I and III).
4* It is possible that the low incidence of trypanosomiasis
among elderly persons may be due, in part at least, to an immunity
acquired by them.
REFERENCES
1. Todd, John L., and Wolbach, S. B. 4 The Diagnosis and Distribution of Human Try¬
panosomiasis in the Colony and Protectorate of the Gambia.’ Ann. Trop. Med. and
Parasit., 1911, V, No. 2, p. 245. Reviewed, Sleeping Sickness Bulletin, 1911, III,
No. 30, p. 336.
2. Dutton, J. Everett, and Todd, John L. 4 First Report of the Trypanosomiasis Expedition
to Senegambia (1902). Liverpool School of Tropical Medicine, Memoir XI, Liver¬
pool, 1903.
3. Dutton, J. Everett, and Todd, John L. 4 Gland Palpation in Human Trypanosomiasis.*
Liverpool School of Tropical Medicine, Memoir XVIII, Liverpool, 1906.
4. Kinghorn, Allan. 4 Age Incidence of Human Trypanosomiasis.* Sleeping Sickness
Bulletin, 1912, IV, No. 40, p. 356.
5. Kinghorn, Allan. ‘Report on Human Trypanosomiasis in the Western Province and
in the Banda District of the Northern Province of Ashanti.’ Sleeping Sickness
Bulletin, 1911, III, No. 25, p, 138.
6. Sanderson, Meredith. 4 The Human Trypanosomiasis of Nyasaland.’ Trans. Soc. Trop.
Med. and Hygiene, 1912, V, No. 8, p. 298.
7. Hearsey, H. Sleeping Sickness Bulletin, 1912, IV, No. 37, p. 195.
8 . Dutton, J. Everett, and Todd, John L. 4 The Distribution and Spread of Sleeping
Sickness in the Congo Free State, with Suggestions on Prophylaxis.* Liverpool
School of Tropical Medicine, Memoir XVIII, Liverpool, 1906, p. 25.
9. Todd, John L. 4 Duration of Trypanosome Infection.’ Arch. Intern. Med., 1911, April,
VII, pp. 500-505; Sleeping Sickness Bulletin, 1911, No. 29, p. 274.
10. For other references see the indexes of the Sleeping Sickness Bulletins.
321
THE IDENTIFICATION OF THE PATHO¬
GENIC ENTAMOEBA OF PANAMA*
BY
S. T. DARLING, M.D.,
CHIEF OF LABORATORY, ISTHMIAN CANAI. COMMISSION, ANCON, CANAL ZONE
(Received for publication 31 March , 1913)
INTRODUCTION
The identification of the entamoeba that causes dysentery and
liver abscess in Panama has recently been placed on a satisfactory
basis, since two of the members of the Hospital Staff have given
special attention to the subject and have had referred to them for
diagnosis all cases of entamoebic dysentery. Formerly each doctor
diagnosed the cases in his own ward, and there was no uniformity
in the identification of entamoeba, .each man determining for
himself whether in a given case the entamoeba was E. coli or
E. histolytica.
Influenced by the work of Schaudinn and Craig, until a year
ago I regarded all pathogenic forms as E. histolytica. In August,
1911, a careful study of the entamoebae found in clinical cases of
entamoebic dysentery and entamoebic liver abscess was begun, and
observations since then have led me to be of the opinion that the
only pathogenic form in this region is E. tetragena , Viereck.
Having ample facilities for the collection of material from
clinical cases, autopsies and the operating room, and for animal
experimentation, attempts were made to determine the specific
characters of the entamoebae so collected.
METHODS OF EXAMINATION
These were: —
1. Fresh and moist-chamber preparations and those stained
intravitam with Gentian violet.
2. Wet-fixed preparations stained with haematoxylin,
Romanowsky modifications and various other stains.
* Read at the XV International Congres* on Hygiene and Demography, Division I,
Section I, September 26, 1912, Washington, D.C.
322
3. Dry-fixed preparations stained by modifications of the
Romanowsky method.
4. Animal inoculations and feeding experiments.
EXAMINATION OF FRESH PREPARATIONS
It was found that very few of the published descriptions of
entamoebae were sufficiently helpful as guides for the purposes of
identification, in fact every step required to be specially worked out,
and clinical and morphological data correlated with animal
experimentation.
Take the questions of refractility of the ectoplasm, and the
colour of the cytoplasm, it seems to me that refractility is very
largely a question of the age of the entamoebae or its relation to
encystment without regard to species. I have found in cultures of
free living forms that the younger amoebae were not very refractile,
while those approaching the period of encystment were quite
refractile. In a fatal case I found that large trophozoites deep in
the submucosa in the floors of ulcers were very slightly refractile,
while many of the smaller entamoebae in the more superficial
sloughs were highly refractile, yet they were representatives of one
species. In some clinical cases, all the forms seen were highly
refractile, many contained chromidia and were the precursors of the
small generation, from which the cysts develop. In other cases all
the entamoebae were finely granular, contained no erythrocytes and
were not refractile. I have never seen either with natural or
artificial illumination the green or grey tints noted by several
writers, and it does not seem to me that stained specimens bear out
the notion that the green colour is due to lysed erythrocytes, for
the latter ar^held intact for a considerable period in the endoplasm
and disappear by condensation and erosion, yet retaining their
staining characters. I should say that all observations were made
with artificial light, transmitted through blue glass, and I wish to
call attention to the necessity of using some such uniform method
of illumination, for the difference in illumination between light
derived from a blue sky or from white clouds makes their use as
sources of light uncertain, and gives inconstant pictures usually not
3*3
noted by the observer. The resulting errors may be eliminated by
the use of artificial light.
The nucleus is said to be inconspicuous in the pathogenic
entamoebae in fresh preparations, but I have not always found this
to be the case when artificial light has been used. It is true that in
many individuals, or in all the individuals in some cases, the
nucleus cannot be made out in fresh preparations. But when one
learns to recognise the nucleus, nothing within the trophozoite is
more conspicuous. The refractile granules or masses of peripheral
chromatin stand out with great prominence, and may be followed
with ease as the entamoeba moves.
The presence of an ectosarc is very constant in the trophozoites
of the pathogenic form, and is a very important feature for
differentiation from E. coli.
While in most cases it is true that all entamoebae detected in
stools associated with pus, blood and mucus are pathogenic, it is not
this type of case that presents difficulties. The puzzling cases are
those in which entamoebae are detected in fluid stools containing
mucus or in semi-solid or formed stools unassociated with blood or
pus, but which it is necessary to diagnose at once so that treatment
may be started energetically. As an aid to rapid diagnosis, I have
used with some success gentian violet as an intravitam stain. The
stain is used in the concentrated fluid form or diluted with an equal
amount of water, and is either added to the flake of mucus or drawn
under the coverslip and the preparation examined immediately.
While many individuals become over-stained or remain under¬
stained, usually they will gradually take up the stain in such a
manner that the peripheral chromatin, the centriole and karyosome
in E. tetragena stand out with almost the clearness and definition
of well-stained haematoxylin preparations fixed in the wet way, and
differentiation from E. coli is easy. I have tried several basic
stains, but none is so good as gentian violet for intravitam staining.
Even when the trophozoite is very refractory to the stain, as in the
small generation, and in the forms which contain chromidia, the
nucleus for some reason will become more refractile and conspicuous.
The non-refractile trophozoites take up the gentian violet rapidly,
while cysts and the refractile forms take up the stain slowly.
EXAMINATION OF STAINED PREPARATIONS
Haematoxylin stained preparations were made from those fresh
films which contained a sufficient number of entamoebae to warrant
staining and study. The coverslip was removed and both slip and
slide were fixed in Schaudinn’s bichloride alcohol, or in Zenker’s
solution diluted one-fourth and one-eighth, as the full strength
solution causes artefacts in the nucleus. Several haematoxylin
stains were used, and particularly clear pictures were obtained with
Mallory’s phosphotungstic acid haematoxylin, with the modification
that the films were not given a preliminary treatment with oxalic
acid or permanganate but were placed over night in the
haematoxylin and then differentiated with very dilute perman¬
ganate. This is a shorter way of using phosphotungstic acid
haematoxylin and gives excellent results.
Fresh and wet-fixed preparations were controlled or compared
with others stained after dry fixation by some modification of the
Romanowsky stain. The films were thoroughly dried then stained
with Hasting’s stain, over-stained with Giemsa’s stain, and
differentiated with ammoniated alcohol. It was observed that the
descriptions of pathogenic entamoebae, as well as figures used to
illustrate them in the literature, did not correspond with the results
obtained in well-differentiated films stained by the method just
described. And it is believed that descriptions in the literature
based on films stained in the usual way, that had not been
sufficiently differentiated, are not as exact as they might be.
Romanowsky stains have a tendency to over-stain just as
haematoxylin does, though not so intensely as the latter, and this
over-staining must be corrected by the use of a differentiating agent.
It has been my practice to stain in the following way. Fresh
coverslip preparations containing a sufficient number of entamoebae
to warrant staining and study, or those intended for diagnosis, are
made into smears by sliding off the coverslip and thoroughly drying
both slip and slide, after which each is stained with Hasting’s stain
for 15 minutes. Satisfactory films are then over-stained with
Giemsa’s stain until the film has a diffuse reddish purple tint. The
film is then plunged into 60% ethyl alcohol containing about
1 % of water of ammonia (10%), and differentiated in this,
3 2 5
washed in water, and controlled by the microscope until the purple
substance of the nucleus and the blue colour of the cytoplasm are
strongly contrasted. The film when properly differentiated has a
blue-violet colour. If the film has been gTeatly over-stained it is
treated with a momentary douche of 95% alcohol. Beautiful
pictures are obtained in this way, but what is of more importance,
the various figures displayed by the purple staining substance
(karyosome?) can be noted and followed with ease. This purple
staining substance in the nucleus of Entamoeba tetragena in dried-
fixed films represents only a portion of the nucleus, as the centriole
and peripheral chromatin do not stain purple by the above method.
The purple staining substance in the nucleus of E. tetragena
frequently appears as a ring, or as a reticulum or scattered granules.
Its phases do not appear to have been accurately described, but
have been confused with the appearances presented in wet-fixed films
when basic stains have been used.
Here I wish to call attention to certain errors of interpretation
which I believe have resulted from the failure to properly
distinguish between the purple staining substance or karyosome of
the nucleus in Entamoeba tetragena in dry-fixed films, and the
nuclear substance which stains with basic stains in wet-fixed
preparations. For example, figures showing the purple chromatin of
the nucleus of E. histolytica represented by Craig and illustrated
in its proper colour, have been used by other writers and figured in
terms of haematoxylin or black and white, without explaining that
the black in the figure represents the purple staining substance of
dry-fixed Romanowsky preparations. Dry-fixed stained films
should never be confused with those stained after wet fixation, for
the pictures are different in each case, and the mistake should never
be made of attempting to translate one into the other. In dry-
fixed films the permeability of the entamoebae to stains apparently
has been profoundly modified, for the centriole and the peripheral
chromatin do not take or retain the purple stain. The karyosome
alone retains the purple stain, while the remainder of the nucleus
stains faintly blue, sometimes revealing the achromatic granules of
uniform size which appear to form part of its structure.
326
FEEDING AND INOCULATION EXPERIMENTS
A number of kittens, cats, dogs and monkeys have been fed by
mouth, or inoculated per rectum with cysts and trophozoites of
Entamoeba tetragena . When tetragena cysts have been fed to
half-grown cats there has resulted not the typical entamoebic colitis,
such as is usually described in the literature, but an enteritis,
and in this lesion in the ileum trophozoites have been found which ,
though arising from tetragena cysts , had the morphology not only
of E. tetragena but of E. histolytica and E. nipponica . Now as a
culture, which from its history and microscopical appearance was
certainly a pure culture of E. tetragena cysts, was used in the
experiment, and as the various forms appeared in the cat’s intestines,
I am led to believe that the trophozoites, described as E . histolytica
and E. nipponica by various writers, are nothing more than the
large trophozoites in the first place and atypical or degenerate forms
of E. tetragena in the latter. I am confirmed in this opinion by
never finding in my cases any of the perpetuating forms described
by Schaudinn and Craig for E. histolytica . In one fatal case of
tetragena infection, I observed from autopsy material many
trophozoites that protruded pseudopodia not unlike those figured
by Hartmann and Craig for E . histolytica . Their extremities were
refractile, and appeared to contain a round spore-like body, yet,
when these coverslip preparations were fixed immediately and
stained with haematoxylin and by Romanowsky, the picture
presented was that of E. tetragena , and the spore-like bodies had
disappeared. In studying a strain of E. tetragena in cats,
following rectal injection of trophozoites, I found at the fourth
remove in dry-fixed preparations a great many trophozoites, the
peripheries of which contained one or several lobose projections,
the interior of which were very frequently deeply stained blue and
suggested strongly the descriptions of E. histolytica by Craig. But
these were artefacts, for in wet-fixed haematoxylin preparations
from the same coverslip preparation the picture was that of
E. tetragena , and associated with these trophozoites were several
uninucleate tetragena cysts. If only the dry-fixed films had been
studied in this case, they might easily have been described as
E. histolytica.
327
With reference to rectal inoculation of trophozoites, infection
practically always follows in animals of the right age, and death
occurs within a few days (five to eight) if the strain is not too aged,
while by mouth feeding the duration is longer (twelve days). In
this, my experiments have not paralleled those of other writers. It
is difficult to explain this. I may say that after several unsuccessful
attempts to infect very young kittens and adult cats, I subsequently
only used kittens weighing about 700 grams.
Animal experimentation is of very great value in studying the
variations in a given strain. In a strain recently I have been able
to watch senility gradually developing from week to week, the
entamoebae becoming reduced in size and filled with chromidia,
and ultimately becoming encysted.
The comparative study of staining and fixing agents has brought
out some very interesting information. The very marked differences
presented by dry-fixed and wet-fixed stained preparations have
been referred to. Phosphotungstic acid haematoxylin gives varying
pictures depending on the fixative. More information of the
structure of the nucleus is obtained by the use of this stain after
fixing with diluted Zenker's fluid (1/8 and 1/4) and Flemming’s
fluid. The karysome is not so well transfixed when treated with
Merkel’s, Hermann’s or Schaudinn’s fluid. In wet-fixed films
stained by Romanowsky it was never possible to stain the
trophozoites exactly like the tissue cells near by. When stained by
Giemsa’s method and differentiated in acetone xylol the peripheral
chromatin usually stained blue, while the centriole and nuclear sap
stained pink or red, the tissue cells near by displayed purple
staining substance throughout the nucleus. It was possible in
several cases to study first the tissue-destroying trophozoites, later
the small generation, and finally during convalescence and after
apparent recovery the cysts, although in practically all cases that
received energetic medication the small generation and cysts did
not appear.
The detection of cysts in convalescent and recovered cases is
most important, for it is the cyst, and not the trophozoite, which
is the infecting agent and makes the host of the former a ‘ carrier.’
The identification of tetragena cysts is usually easy in stained
preparations and should present no difficulties in fresh films, yet
3*8
the cysts are so small (12 to i$/k in diameter) that they have
frequently been mistaken for monad or coli cysts or fat droplets,
or possibly mononuclear leucocytes. When in doubt and the
number of cysts was to small to risk loss by fixation and staining,
I have vaselined the preparation or kept it in a moist chamber, and
if the cyst was homogeneous at first, after one or two days, one,
two or four nuclei became distinctly visible. Tetragena cysts are
more commonly detected in neglected cases which have partly
recovered from entamoebic dysentery who may have diarrhoea or
whose stools are solid, and I have found cysts in a case that had
been insufficiently treated by means of rectal injections. It would
seem, however, that if a case of dysentery is treated early and
energetically the trophozoites are at once driven from the field,
leaving none to develop into the drug-resistant small generation
from which the cysts arise. This, it will be seen, is analogous to
the rational treatment of malaria, in which the asexual generation is
destroyed at once by large doses of quinine, thus destroying all the
forms from which the gametes arise.
In acute or new infections, or in very active lesions, many of the
trophozoites are of large size—30 to 6o/» in diameter. As the strain
grows older, the size often becomes reduced, the trophozoites
measuring from 12 to 24/1 in diameter. These forms frequently
contain coarse blocks of chromidia, and they constitute the ‘ small
generation.’ During convalescence and after apparent recovery
there appear small trophozoites 12 to 15* in diameter, and
associated with them are cysts and four-nucleated schizonts. If
relapse occurs with symptoms of colitis, large trophozoites make
their appearance again, and the cysts will have disappeared. Thus,
during the progress of a case of tetragena dysentery, at first large
trophozoites will be seen, many or all of them having the characters
described for E. histolytica by Schaudinn and Craig, and later if
the case has been a neglected one, the small generation with cysts
will be found, of which there is no better description extant than
that in Elmassian’s paper on this form, which he has called
E. minuta.
3 2 9
CONCLUSIONS
If there is a sufficient number of cysts they may be fed by mouth
to young cats, and it may be possible to recover, from their bowel
lesions, trophozoites having not only the characters of E. tetragena ,
but of E. histolytica and E . ttipponica as well. Or, if in any given
case of tetragena dysentery, in which the trophozoites are of the
histolytica type, they be injected rectally into a young cat and the
strain carried on by subsequent rectal inoculation into other cats at
the time the infected animal dies, so as to prolong the vegetative
phase of division, then it will be seen that the histolytica- like
trophozoites become reduced in size, filled with chromidia, and at
the fourth or fifth remove it is possible to find uninucleate
tetragena cysts. The nucleus of the trophozoites meanwhile has
taken on a typical tetragena appearance with a prominent
karyosome. It is now impossible to infect other cats per rectum
with this material. If we make dry-fixed Romanowsky stained
smears of material containing these typical tetragena trophozoites,
they will occasionally present the morphological peculiarities of
‘ E. histolytica 9 described by Craig. We are thus able to correlate
most of the observations of Schaudinn, Craig, Elmassian and
Koidzumi, and state that there is but one pathogenic entamoeba,
and that one is E. tetragena .
REFERENCES
Ckaig, C. F. (1908). ‘ Studies upon the Amebae in the Intestine of Man.’ Journ. Infect.
Diseases, V, pp. 324-377, 2 plates.
Elmassian, M. (1909). ‘ Sur une nouvelle esp£ce amibienne chez l’homme, Entamoeba
minmta , n. sp.* Centralbl. f. Bakteriol., Abt. i, Orig., LII, pp. 335 - 351 , 2 plates.
Haktmann, M. (1908). ‘Eine neue Dysentericamobe, Entamoeba tetragena (Viereck), syn.
Entamoeba ajricana , (Hartmann).* Arch. f. Schiffs- u. Trop.-Hygiene, Bd. XII,
Beiheft 5, pp. 117-127.
- (1909). ‘ Untersuchungen iiber parasitischen Amoben. I. Entamoeba histolytica
Schaudinn.* Arch. f. Protistenkunde, XVIII, pp. 207-220, 1 plate.
Koidzumi, M. (1909). ‘ On a new parasitic Amoeba, Entamoeba nipponica , found in the
intestine of Japanese,* Centralbl. f. Bakteriol., Abt. 1, Orig., LI, pp, 650-653.
Schaudinn, F. (1903). ‘Untersuchungen uber die Fortpflanzung einiger Rhizopoden.*
Arb. a. d. Kaiserl. Gesundheitsamte, XIX, Heft 3, pp. 547-576 (see p. 563.)
V iukcx, H. (1907). * Studien liber die in den Tropen erworbene Dysenteric.’ Arch. f.
Schiffs- u. Trop.-Hygiene, Bd. XI, Beiheft 1, pp. 1-41, 3 plates.
33 *
A NEW TSETSE FLY FROM THE CONGO
FREE STATE ; AND THE OCCURRENCE
OF GLOSS IN A AUSTEN I IN GERMAN
EAST AFRICA
BY
Professor R. NEWSTEAD, F.R.S.,
THE I.IVBRPOOL SCHOOL OF TROPICAL MEDICINE
{Received, for publication 3 April, 1913)
Recently Monsieur G. Severin, of the Mus£e Royal d’Histoire
Naturelle de Belgique, had the kindness to send me an extensive
collection of tsetse-flies for identification, numbering in all nearly
one thousand specimens, all of which had been collected in various
parts of the Congo Free State. Among these were found two
examples of a species which has proved to be new, and hitherto
undescribed. I wish here to express my indebtedness to Monsieur
Severin for entrusting me with the examination of the collection in
question, and also to add that it affords me infinite pleasure to be
allowed to dedicate this new species of Glossina to so distinguished
a naturalist.
Glossina severini, sp. n.
MALE. Hind tarsi either uniformly dark brown or with the
first and second segments slightly paler than the rest; pleurae and
hind coxae dusky grey, harpes (fig. 1 h) very small, narrow and
irregularly serrated distally.
Length (two specimens) 10 3 to 10 5 mm. Length of wing
iO'5 mm.
Palpi relatively long and stout. Third segment of the
antennae clothed with short hair. Thorax very robust, markings
well defined and of the usual type or similar to those
in Glossina pcdpalis; pleurae dusky grey. Legs with the hind
coxae grey or greyish buff; tips of front and middle
tarsi black; hind tarsi all dark (almost uniformly so in one
example), or with the first and second segment slightly paler than
the rest. Abdomen almost unicolourous. Genital armature (fig. 1)
332
with the superior claspers free, the hairs with which these
appendages are clothed very long. Editum with very long
marginal hairs, the longest reaching almost to the tip of the vesica
( y ); median process (mp) narrow, being much less than the width
of the inferior claspers (i. c .) and projecting slightly beyond the
latter; harpes (h) rudimentary, being quite short and narrow, with
the distal margin irregularly serrated.
Fig. i. Glosstna sever ini, Newstead.
The type specimen of this species is a male in the Mus6e Royal
d’Histoire Naturelle de Belgique. It was taken in the Congo Free
State, and bears the following data: —‘ Lac Mouro (Riv. La Lavua),
Mai, 1907.’ It had evidently been preserved in alcohol, so that it
is highly probable that the colours may have been slightly changed;
but on comparing it with a number of examples of Glosstna
fuscifleurts, Aust., from the Congo Free State, and also some
specimens of Glosstna fusca, Walk., all of which had been
preserved in the same way, one could readily separate both
G. severmt, n.sp., and G. fuscipleuris from G. fusca by the much
darker colour of the pleurae and the hind coxae.
A second example from the same Institution as the type bore the
label ‘ Congo ’ only.
As to the affinities and characteristics of this new tsetse-fly, one
gathers from the two examples that, so far as the colour of the hind
coxae and pleurae are concerned, it resembles G. fuscipleuris; but
it has relatively longer and stouter palpi; and furthermore is
distinguishable from the latter by the darker and more uniformly
coloured hind tarsi.
If we take the structural characters of the male genital armature,
we find that it is very closely related to G. medicorum, Aust., from
which it may, however, be readily separated by the curious rudi¬
mentary harpes, the greater length of the marginal hairs of the
editum, and furthermore by the greater length of the lateral
branches of the hairs of the arista (fig. 2 a).
Fio. 2. (a) Gloss ina srotrini , n. ip. Terminal portion! of the hairs of the arista.
(b) Glosstna wudtcorum, Austen. Terminal portions of the hairs of the arista.
Soon it is hoped that perfectly fresh material will be available
for a more complete study of the colour and pattern of this new
334
tsetse-fly, as too much importance cannot be attached to the colour
in the existing specimens, owing to the fact that changes, as already
stated, may have taken place. However this may be, the structural
characters as set forth in the above diagnosis will very readily assist
in the future determination of this interesting addition to the genus
Glossina .
Glossina austeni , Newstead.
Twelve examples of this recently discovered tsetse-fly, consisting
of two males and ten females, were submitted to me for identification
by Dr. Hermann Morstatt, Amani. The examples in question were
captured at Nyussi, Tanga district, January and February, 1913.
Though this is the first recorded instance of the occurrence of
this species in German East Africa, its occurrence there is not
surprising seeing that it has also been recently recorded from
Portuguese East Africa. Its northernmost range, so far as is known
at present, is Jubaland. It is evident, therefore, that this insect
has a wide geographical distribution, and it is remarkable that so
distinctive a species should have escaped detection until quite
recently.
335
EXAMINATION OF THE ROOT OF AN
IPOMOEA FROM RHODESIA
BY
PROSPER H. MARSDEN
LECTURER IN MATERIA MID1CA AND PHARMACY IN THE UNIVERSITY OP LIVERPOOL
(Received for publication 16 March , 1913)
PLATE XXVII
I am indebted for the specimen of the root of Ipomoea to
Dr. W. Yorke, who brought it from Rhodesia. Dr. Yorke says of
the root: ' It is the native remedy for a disease known as Chilatera,
which has quite erroneously been confounded with Sleeping
Sickness. A native doctor who visited our Sleeping Sickness
Camp in the Luangwa Valley stated that several of the patients
were suffering from Chilatera, and that he could cure them by
administering this drug. On obtaining our consent he made a
decoction of the root and administered it to the patients in question.
Within an hour they all exhibited marked signs of gastro-intestinal
irritation with acute vomiting and diarrhoea. I think that, as the
drug has such a powerful cathartic and emetic action, it would be
of interest to attempt to ascertain the active principle.*
The material consisted of a rough tuberous root (PI. XXVII)
weighing 260 grammes, 20 cm. long and 24 cm. in diameter,
having the external characters of that of Ipomoea horsfalliae
described by Power and Rogerson.*
MACROSCOPIC AND MICROSCOPIC CHARACTERS OP ROOT
The root was found to consist of a hard outer portion, of an
ashy-grey colour, with warts upon it and exhibiting dark spots of
exuded resin. The inner portion, evidently decomposed, showed a
black pulverulent mass which was interspersed with greyish vessels.
• Wellcome Laboratories Report No. 171, reprinted from the American Journal of
Pharmacy, August, 1910.
336
Upon examining the latter material under the microscope the vessels
were seen to have thickened walls, and starch grains, simple and
compound, were present in large quantity. The outer portion
consisted of some ten layers of yellowish-brown cork cells, and
resin-secreting cells were seen in the cortex and the bast, whilst
starch grains were numerous in the parenchyma.
PRELIMINARY EXAMINATION
Upon digesting a small quantity of the inner portion of the root
with water, in a water bath, for two hours an extract was obtained
which was alkaline to litmus, gave no reaction for tannin or
alkaloids, but reduced Fehling's solution, and gave a slight cloud
with solution of acetate of lead, and a persistent froth upon
shaking. An extract of the outer portion, prepared in a similar
way, was acid to litmus, reduced Fehling's solution, and frothed
upon shaking.
<**
ESTIMATION OF MOISTURE
The moisture, estimated upon 14*29 grammes of whole root, was
found to be 61*47 P^ r cent.
EXAMINATION FOR VOLATILE OR FIXED OIL
As the root had a slightly aromatic odour it might contain a
volatile oil; with a view of ascertaining this, 4*2 grammes of the
finely-powered root was mixed with twice its weight of fine sand
and exhausted in a Soxhlet apparatus and reflux condenser with
petroleum spirit. Upon evaporation of the latter a syrupy liquid
was obtained corresponding to 2*59 per cent, of the material taken.
This quantity was too small to make much of, and upon passing
steam over it in a closed vessel no aromatic odour was obtained.
EXAMINATION FOR PRESENCE OF RESIN
60 grammes of the dried root was cut up, and separated
into outer and inner portions. These were present in the proportion
of three of inner to one of outer. Each was powdered, mixed with
twice its weight of fine sand and exhausted with hot alcohol in a
Soxhlet apparatus. Most of the alcohol was recovered by
distillation, and the remainder evaporated until the extract ceased
337
to lose weight. In the case of the outer portion the result was a
soft extract corresponding to 6*95 per cent, of the material taken,
and having an odour of extract of gentian. This extract was
digested with water on a water bath, cooled and filtered; the filtrate
was slightly acid to litmus and gave no reaction for tannin or
alkaloids, nor did it reduce Fehling’s solution. A dark green resin
was left upon the filter paper corresponding to 0 85 per cent, of the
dried outer portion of the root taken.
The inner portion of the root treated in a similar way yielded
8* 19 per cent, of soft extract and 370 per cent, of resin.
I am much indebted to Messrs. G. B. Westmacott and Son, who
very kindly procured from Jamaica for me a fresh root of Ipomoea
horsfalliae for purposes of comparison.
Dr. Yorke is endeavouring to procure leaves and flowers of the
plant which yields the root I have described above; until these
arrive the identity of the drug must remain uncertain.
338
EXPLANATION OF PLATE XXVII
Tuberous root of lpotnoea horsfalltae —a native remedy for the
disease known as Chilatera, in Rhodesia.
A.
Volume VII
August, 1913
No. 3A.
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Edited by
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
AND
Processor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.,
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University,
Liverpool.
C. Tilling &* Co., Ltd.
Printers to the University Press of Liverpool
53 Victoria Street
339
ON THE MORPHOLOGY OF THE
TRYPANOSOME (T. NIGERIENSE, n. sp.)
FROM A CASE OF SLEEPING SICKNESS
FROM EKET, SOUTHERN NIGERIA
BY
J. W. SCOTT MACFIE, M.A., B.Sc.
MEDICAL RESEARCH INSTITUTE, LAGOS, SOUTHERN NIGERIA
(.Received for publication 25 July , 1913)
Early in 1912 attention was drawn to the presence of a disease
resembling sleeping sickness in the Eket district of Southern
Nigeria. Mr. S. A. Bill, of the Qua Ibo Mission, reported in
February that a girl at a village called Ikot Offiong appeared to be
suffering from trypanosomiasis. A medical officer (Dr. R. W.
Gray) was accordingly despatched to the district to investigate this
case, and to enquire into the presence of sleeping sickness in the
neighbourhood of Eket. In May he was able to report that he had
discovered two cases; and, as on further investigation the disease
was found to be widespread, an isolation camp was established near
Ikorobo. Up to the end of April, 1913, a total of 167 cases of
sleeping sickness had been identified.
Sporadic cases of sleeping sickness are known to occur in many
parts of Nigeria, and in certain districts the disease appears to be
endemic. The writer has seen cases at Baro and Kateri in Northern
Nigeria, but although trypanosomes were present in the peripheral
blood of these patients, they did not appear to be ill. In Kabba
province of Northern Nigeria it is said to be endemic, and along
the course of the Garara river, where the writer had an opportunity
of investigating the subject in 1910, it was well known, but
apparently uncommon. Sleeping sickness does not seem to be of a
virulent type in Nigeria, and it does not at present occur in epidemic
form. To account for this fact it has been supposed that the
disease must have existed for a great number of years, and that the
natives must have acquired a relative immunity. Some such
340
explanation is necessary since sporadic cases have been identified
in so many places, and the local conditions over so much of the
country are so favourable for the spread of the disease.
Sleeping sickness as it occurs at Eket, however, presents some
special features. It was thought, therefore, that a study of the
trypanosome producing the disease might be of interest. For this
purpose Dr. Foran, the medical officer in charge of the sleeping
sickness investigations, very kindly sent the writer a guinea-pig
which had been inoculated from one of his cases. On examination
the trypanosome was found to possess some unusual morphological
features which are, I think, of sufficient interest to place on record,
in view of the peculiar clinical symptoms with which they are
associated.
SLEEPING SICKNESS IN EKET DISTRICT
I am indebted to the official reports of Dr. P. F. Foran for the
following account of the disease as it occurs at Eket.
Sleeping sickness has, apparently, been known to the natives of
the district for a very long time. It has, indeed, existed beyond
the memory of the oldest inhabitants. They state, however, that
it has become more common lately, and this they attribute to the
fact that the present generation is inclined to ignore certain old
native laws that forbade an infected person to associate with his
fellows. Two phases of the disease are recognised. The early
phase, characterised by enlargement of the glands of the neck, is
known as nsipiton; and the later phase, in which lethargy appears,
is called odongo-idap* Cases in the former stage are common, but
those that have advanced to the latter stage are comparatively rare.
The disease appears to be mild, although a number of deaths are
reported to occur annually. In September, 1912, Dr. Foran wrote,
* I have only heard of four deaths from sleeping sickness, yet
* The writer, in 1910, found that a similar distinction was observed by the natives of
Northern Nigeria. The stage of glandular enlargement they termed cbttoan toiya (neck
sickness), and the stage of lethargy cbttoan bericbi (sleep sickness). Sleeping Sickness Bureau
Bulletin, No. 27, p. 236.
3+t
nsipiton [the early, glandular, phase of the disease] is very
common/ The duration is a matter of years. In the cases
collected, symptoms had been present for from one to five years.
The great majority of the patients are children or young adults,
their ages ranging from about 6 to 18 years. The sexes are about
equally affected.
The symptoms are as follows: As a rule the general health
is good, there is no marked anaemia, and wasting is present in only
a few cases. Irregular fever occurs, especially in the afternoon and
evening, and is accompanied by headache and malaise, and
terminates in sweating. The tongue is furred. The spleen is
almost always enlarged. The expression of some patients is dull
and vacant; and there may be oedema or drooping of the eyelids.
Muscular tremors occasionally occur. Weakness or lassitude is
always more or less well marked. Skin rashes—urticaria and
erythema—are present in many cases; but they are also common in
uninfected natives. Impotence and amenorrhoea are said to
accompany the disease. Enlargement of the cervical and axillary
glands is the most constant sign. The glands are freely movable,
soft, and elastic; and on microscopical examination are found to
contain trypanosomes. Enlarged glands in other sites, such as the
groin, do not contain the parasites. On excision, the glands are
found to be pale pink in colour. They do not contain pus, and it
is noteworthy that those showing signs of induration are found to
be free from trypanosomes. The natives believe that the disease is
curable by the excision of the glands, an operation which their
1 doctors ’ readily perform. Numerous healthy individuals bearing
scars on their necks are to be met with who have, it is affirmed, been
cured in this manner** According to the natives the glandular
phase of the disease ( nsipiton) lasts for about four years before
lethargic symptoms develop. From the fact that somnolence is
not a common symptom, most of the patients appearing to be in
the early stages, it is possible that many of them recover before
this stage is reached.
Trypanosomes are found in the gland juice of practically every
* Compare the note on the excision of cervical glands by a native doctor in Kabba
province, Northern Nigeria. Dr. W. Morrison. British Medical Journal, June 8th, 1912.
3+2
case, but, up to the present, they have not been detected in the
peripheral blood. In a few cases presenting the appearances of
advanced sleeping sickness trypanosomes have not been found at all.
In them the glands are but little enlarged, and, as noted above,
indurated glands are generally free from parasites.
The country around Eket is an undulating plateau covered with
dense bush, and intersected by numerous waterways. It includes
a good deal of swampy land. The district is densely populated.
The towns consist of scattered compounds standing some distance
apart, and closely surrounded by farms and banana trees. They
are usually very dirty, and some cattle, and a good many pigs,
goats, and fowls are kept in them. The towns are generally some
distance away from the water-side. Their water supply is obtained
at two or three spots, which are closely surrounded by bush and
trees, and approached along a narrow shaded path. The children
and young adults are the water-carriers, and Dr. Foran considers
that this accounts for the majority of the cases of sleeping sickness
occurring in young people. Tsetse flies are prevalent all over the
district. Dr. J. J. Simpson records G. palpalis and G. caligtnea
from Eket in his map of Southern Nigeria, showing the distribution
of the genus Glossina;* but, according to Dr. Foran, G. tachinoides
is also a common species, at any rate, during the months of
September and October, t He has observed that the tsetse flies
‘ appear to follow pigs about more than any other animal, and it
is generally easier to catch the flies where these animals are than at
the water.’
THE STRAIN OF TRYPANSOME
The trypanosomes used in this investigation were found in the
blood of a guinea-pig kindly sent to the writer from Eket by
Dr. P. F. Foran. The guinea-pig had been inoculated with
cerebro-spinal fluid from a case of sleeping sickness on March 5th,
1913, but the incubation period is not known. The animal reached
Lagos on May 8th, 1913, and was found to be infected with
* Bulletin of Entomological Research, Vol. Ill, part 2, 1912.
f If true, this is a remarkable fact as G. tachinoides does not occur so near to the coast
elsewhere in Southern Nigeria. It is generally only met with inland, beyond the forest rone.
3+3
trypanosomes. It appeared to be well, and, at the time of writing
(June 11th, iqi 3), is still in good health, although it is now 97 days
since the date of inoculation. The trypanosomes have never been
very numerous in the blood. Their numbers have not sensibly
increased during the last month. It is, therefore, impossible to tell
to what stage the disease has at present advanced.
The red blood corpuscles of the host exhibit well marked
polychromasia and basophilia. In many of them, too, irregularly
shaped fragments are to be seen, which stain red with Gimesa's
solution.
MORPHOLOGY OF THE TRYPANOSOME FROM EKET
In this paper the flagellar end of the trypanosome will be termed
anterior, and the non-flagellar end posterior.
A— Living , unstained .
The trypanosome from Eket in the fresh condition, as seen in a
drop of blood from an infected guinea-pig, appears as an elongated
tapering body of almost homogeneous consistency. The micro¬
nucleus can sometimes be distinguished as a small refractile body.
The movements are active. Some individuals, which appear to be
stouter than the others, are relatively sluggish, and do not move
actively about the field. Others, which are long and slender,
vibrate exceedingly actively, and also move rapidly across the field
of the microscope. The translatory movement seems to be
spasmodic, the trypanosome suddenly gliding across the field after
having been more or less stationary, but vibrating vigorously all the
time, for some moments previously. As a rule, the flagellar end
moves forwards in the translatory movements. The non-flagellar
end appears to be blunt, the flagellar attenuated. The undulating
membrane is conspicuous.
In blood that has been shed for some little time changes may be
observed taking place in the form of some of the trypanosomes.
The writer has watched these changes on several occasions. They
always occurred in long slender parasites, and were accompanied by
a diminution of the activity of the trypanosome. The non-flagellar
end of the trypanosome appears to round off, and eventually the
344
body becomes almost spherical. These atypical parasites are
occasionally met with in stained blood films. One is illustrated
(Plate XXXIII, fig. 22).
B —Fixed and stained .
Method of fixation and staining.—The blood films were fixed
either with absolute alcohol or with osmic acid. The forms of the
trypanosome were found to be the same in either case, but with
osmic acid fixation the appearance of the cytoplasm was different
if the films were stained very deeply. All the films were stained
with Giemsa’s solution (about 30-35 drops in 10 c.c. of water); those
fixed with alcohol being stained for one hour, and those fixed with
osmic acid for from ten to fifteen minutes.
The longest trypanosome measured, up to the present, is 32/i,
and the shortest 8 fi.
Breadth .—The breadth of the trypanosomes at the widest part
varies from ifi to 2 5 /*.
Shape .—The trypanosome when stained is seen to be poly¬
morphic. Long slender forms, short stumpy forms, and
intermediate forms are always present; but their relative proportions
vary considerably from day to day. The trypanosomes have never
been very numerous, and, as a rule, the long slender forms have
predominated. The long slender forms are of two types. In the
one type the blepharoplast is terminal, and the nucleus elongated
(Plate XXXIII, fig. 1). In some specimens the nucleus appears to
have a clear area in the middle which gives it the appearance of
being made up of two pieces, and the body anterior to it is often
attenuated. In the other type the blepharoplast is situated about
1 fi from the posterior extremity, which is prolonged beyond it into
a blunt snout (Plate XXXIII, figs. 2, 3). The intermediate forms
are very variable in size and general appearance. Some are pointed
at both ends, others have a blunt posterior extremity, which is
occasionally rounded off like the head of a tadpole. The
blepharoplast is either terminal or sub-terminal. The body ends
anteriorly in a very short 1 free * flagellum. The stumpy forms are
exceedingly short in some instances (Plate XXXIII, figs. 11, 12),
and may measure as little as 8^1, They are relatively very broad,
345
measuring about 2 ft. At the anterior end there is a very short ‘free *
flagellum. The membrane is not broad. The posterior end is quite
blunt, and the beginning of the flagellum may form the posterior
border of the parasite. The blepharoplast, as a rule, lies at the
edge of the trypanosome a little way in front of the posterior end
of the body, and on the opposite edge the folds of the membrane
pass forwards. The nucleus is large, and is sometimes placed at
the extreme anterior end (Plate XXXIII, fig. 13).
From time to time, in films prepared in the usual way, there
appear in the blood trypanosomes which have the flagellum free in
its entire length (Plate XXXIII, figs. 14, 15). They have not been
observed in unstained blood-films, and, indeed, they would be very
difficult to make out in fresh preparations. It is, therefore, possible
that they may be artefacts; but it is curious that they have never
been seen before by the writer in any other animal, although he
invariably prepares his blood films in exactly the same way. These
forms are long slender trypanosomes with the blepharoplast situated
some way from the extremity. The anterior end is drawn out into
a filamentous termination. The body measures about 22/1, and the
flagellum 31/t.
Contents of the Cell .—In well stained specimens the protoplasm
is homogeneous in structure, neither granules nor vacuoles being
present. In films that have been over-stained, especially if they
have been fixed with osmic acid, both granules and vacuoles appear
(Plate XXXIII, figs. 20, 21). The granules are coarse, and occur
on both sides of the nucleus. Sometimes they almost fill up the
body of the trypanosome.
The nucleus .—The nucleus is oval, and about 2*5/4 in length.
The nucleus of the long slender trypanosomes with a terminal
blepharoplast has already been referred to. In the stumpy forms
the nucleus is rounded, and is generally surrounded on all sides by
a zone of protoplasm. The nucleus is situated near the centre of
the body in the majority of the trypanosomes, but in a few, and
these are always stumpy forms, it is placed anteriorly—that is, at the
flagellar end (Plate XXXIII, fig. 13). This is an interesting
feature, as it suggests that the position of the nucleus in the
stumpy forms is an index of the virulence of the trypanosome. In
the typical T. gambiense it is central, in the more virulent
34 «
T. rhodesiense it is sometimes posterior, and in this less virulent
strain of Nigeria it is anterior.
Blepharoplast .—The position of the blepharoplast has already
been mentioned. It is well marked, round, and deeply staining.
Undulating membrane .—In the long slender forms the
undulating membrane is ample, and is thrown into a number of
folds. In some it is apparently absent, and the flagellum is, in
consequence, free in its whole length. The membrane is less folded
in the stumpy and intermediate forms. In carefully prepared
specimens the membrane can be followed almost to the extremity
of the flagellum, even in the slender forms which at first sight
appear to have a long 1 free* flagellum (Plate XXXIII, fig. 4).
Flagellum .—The flagellum stains deeply. It is well marked
in all forms, and in all forms the terminal portion is free—that is,
projects beyond the protoplasm of the cell and the undulating
membrane. As already stated, in some forms of* long slender
parasites, in stained preparations, the flagellum appears to be free
in its entire length.
Curve of Measurements : —
In their paper entitled ‘Further measurements of Trypanosoma
rhodesiense and T. gambiense n Prof. Stephens and Dr. Fantham
have pointed out that in measuring trypanosomes it is advisable to
confine observations to a single animal, as it is possible that the size
of the parasite may vary in different hosts. They also recommend
that the trypanosomes should be measured on ten consecutive days
so as to obviate the errors due to the daily variation in the numbers
of short and long forms. They suggest that 100 trypanosomes
should be measured on each of the first ten days of the infection
in a white rat. It has been impossible for me to follow this plan
exactly because no white rats have been available, and, in the few
animals that have been infected by sub-inoculation, the parasites
have been extremely rare at the commencement of the disease, and
have, indeed, been apparently absent altogether from the peripheral
blood for several days at a time. I have, therefore, measured
100 trypanosomes on each of the ten days from the 96th day to the
105th day after inoculation in the original guinea-pig sent to me
from Eket
347
With this exception, I have endeavoured to follow the procedure
of Stephens and Fantham as closely as possible, so that my figures
might be comparable with those given by these authors for
T. rhodesiense and T. gantbiense .
Thin blood smears, dried in the air, were fixed in absolute
alcohol and stained with Giemsa’s solution. The trypanosomes
were then drawn with a camera lucida, and the length of a line
drawn through the middle of each was measured by the ‘ tangent
line’ method. This method is, I believe, the most accurate that
has as yet been described.
In the following tables the results of these measurements are
given in detail, so that a closer analysis than mere averages would
permit may be made by anyone who should wish to do so.
A summary of the measurements is added, and a table showing the
distribution into the stumpy, intermediate, and long form groups
of Bruce.
Table 3 shows the great variation in the number of intermediate
and long forms of trypanosome on particular days. The
variation is not, however, so great in the percentage of stumpy
forms, ranging only between 66 % and 36 %. On comparing this
table with that given by Stephens and Fantham, it will be seen that
there is a lesser degree of variation in the figures for all three groups,
and in particular the number of the intermediate forms is strikingly
less. Intermediate forms, that is trypanosomes measuring from
22fi to 24/1, were indeed comparatively scarce, although actually
they were among the most common lengths of parasite. The
relatively slight degree of variation may have been due to the period
to which the disease had advanced in this animal. The figures
given by Stephens and Fantham referred to a rat during the first
ten days of the disease, those dealt with in this paper were from
a guinea-pig between the 96th and the 105th day after inoculation.
34 8
Table I.—Distribution in respect to length of 1,000 non-dividing individuals of trypanosome from a
single guinea-pig
Day after
inoculation
96th day.
97th day. 6
7
8
9
10
98th day. 11
12
>3
In Microns
100th day. 21
8
9
10
11
12
*3
j '5
16
'7
18
*9
20
21
22
2 3
2 4
2 5
26
2 7
28
2 9
30
3 *
3 2
33
34
Of each
20
1
—
2
2
1
5
4
—
1
1
1
2
5
'
'
1
1
1
2
1
2
I
1
1
18-95
20-15
—
—
-
1
<
1
2
2
2
2
2
2
2
2
2
19-90
1
2
2
2
■
1
2
1
2
1
1
2
18-40
1
,
2
1
1
2
2
2
2
1
2
1
I
19*00
I
1
2
2
4
2
2
2
2
1
■
2275
I
1
2
1
2
3
2
1
2
1
2
1
22-20
2
2
>
1
1
2
3
1
1
I
1
2
2 I- 2 5
...
2
1
3
5
2
2
2
1
1
22-55
...
■
1
2
1
2
4
2
5
1
22-25
2
1
1
1
2
1
2
...
2
2
2
22-35
<
1
4
2
■
P
1
«
2
21-go
,
2
1
,
1
4
1
3
3
1 1
«
* 9*35
...
1
2
2
4
2
2
1
1
2
1
I 9* , 5
...
1
1
L
2
2
3
1
! 1
2
1
3
1
1
19-50
1
1 1
1
2
2
2
I
1
3
2
2
1
2370
_!
1
b
1
2
j
4
3
2
1
23-60
1
2
5
r
1
2
1
1
2
20-85
***
3
| 1
2
1
1
1
3
2
3
1
23-40
1
3
2
—
1
3
2
3
2
2
1
23*40
1
2
p
3
7
7
1
19-80
—
1
...
1 2
3
3
3
1
j.
1
!_
p
1
19-50
1
...
1
I
1
p
3
1
2
,
,
1
4
1
20-85
1
_
2
1
2
2
1
1
5
2
1
20-30
...
2
_3
1
7
T
1
|...
i,
7
1 1
21-10
...
Averages
Of each
100
19-28
20-45
22-99
20-31
L
35 °
Table II.—Summary of measurements (in microns) of the lengths of 1,000 individuals
of the trypanosome from Eket, Southern Nigeria, from a single guinea-pig.
Maximum
Minimum
Averages of
each 100
Averages of
each 20
Range of
a/erages of
each 20
I
3 °
11
18 95
2
3 o
12
20*15
96th day. 3
2 5
*3
19*28
I 9-90
*75
4
2 5
10
O
y
00
5
26
10
19-00
6
2 9
*4
22 *75
7
2 9
H
22*20
97th day. 8
2 9
12
22-20
21*25
9
34
*5
22 *55
10
2 7
*5
22-25
11
3 o
*5
22*35
12
2 9
15
21*90
98th day. 13
28
12
20-45
* 9*35
14
30
8
* 9 * *5
*5
28
9
19*50
■ 6
3 1
12
2370
•7
3 2
12
23*60
99th day. 18
30
11
22-99
20*85
2-85
*9
3 i
16
23*40
20
3 2
16
23*40
21
26
12
19*80
22
26
*5
19*50
100th day. 23
30
10
20-31
20*85
1*60
2 4
2 9
11
20*30
2 5
3 o
*4
21*10
35i
Table IT .—Continued
352
Tabli III.—The trypanosome from Eket, Southern Nigeria, arranged in the three groups—
(a) 21/1 and under; (6) 22-24/1; (c) 25/1 and over.
Day
96
97
98
99
100
101
102
io 3
m
Total*
Stumpy 8-21/1
66
36
5*
39
62
4 i
67
57
5 2 3
Intermediate
22-24/1
21
35
22
18
21
*4
16
8
12
1
190
Long * 5 - 34 M
*3
29
20
B
i
25 j 45
30
287
100
100
100
100
100
100
IOO
100 | 100
IOO
1000
Table IV.—Comparison of the trypanosome from Eket, Southern Nigeria, and 7 . gambtenu
(as given by Stephens and Fantham).
-
Average
!
Maximum
Minimum
Under
22/1
22-24/1
25/1 and
over
7 . gambiense .
ft
OO
36*0/1
16*o/i
ii-i%
* 7 "°%
54 '*%
7 . nigeriense .
2I*486/1
34 *°M
8*o/i
5 2 '3 %
19-0%
2 * 7 %
In Chart I a curve is given representing the distribution by
percentages in respect to length of 1,000 non-dividing specimens of
the trypanosome from Nigeria from a single guinea-pig. The curve
given by Stephens and Fantham for T . gambiense is added on the
chart to facilitate comparison.
The main object of the present investigation was to compare the
measurements of the trypanosome from Southern Nigeria with those
of a typical strain of T . gambiense . The figures have, therefore,
been recorded in a manner that will permit of their being compared
in detail with those given in what is, I believe, the latest exhaustive
study of T. gambiense , that is, the paper by Stephens and Fantham
already referred to repeatedly.
It will be observed that the trypanosome from Southern Nigeria
is somewhat shorter than T. gambiense; the average length of
353
354
l,ooo individuals being 21‘486/i, as contrasted with 24*867/1. The
maximum length of T. gambiense as given by Stephens and
Fantham is 36*0/1, and the minimum 16*0/1, as compared with
34*0/1 and 8 0/1 in the case of the Nigerian trypanosome. The curve,
too (Chart I), is strikingly different from that given by these authors
for T. gambiense. It is lower and more extended, and reaches its
main peak at 21/1, instead of at 26/1. At the far end, representing
the exceptionally long parasites, the two curves almost coincide, but
from this point backwards there is no agreement, and when the
small forms are reached, the Nigerian trypanosome shows a very
much higher percentage than that in the typical strain of
T. gambiense . It has been already pointed out that the occurrence
of these extremely small individuals is characteristic of this strain
of human trypanosome.
ANIMAL REACTIONS
Dr. Foran, at Eket, injected a number of rats with blood and
gland juice from cases of sleeping sickness, but none of them
developed parasites. He used 1 three kinds of rats in the experi¬
ments, namely, a large black house rat, a smaller brown bush rat,
and a brown striped rat’; but ‘all appeared to be immune.’
The animal reactions of the trypanosome are at present under
investigation. It will, however, be some time before anything
definite can be said about them, as the strain seems to be but slightly
pathogenic. The following table embodies the results so far
obtained: —
355
Animal
Days since
inoculation,
up to July 8
Day on which
trypanosomes
were first
detected
Remarks
Guinea-pig
i
124
?
Alive and well. Trypanosomes numerous.
The original animal from Eket.
»
2
56
...
Has never shown trypanosomes. Alive and
well.
16
15
...
Died on 15th day. Never showed trypano¬
somes. Tissues nil.
»»
*7
34
...
Has never shown trypanosomes. Alive and
well.
22
2 4
Has never shown trypanosomes. Alive and
well.
)»
2 3
2 4
Has never shown trypanosomes. Alive and
well.
Mouse
7
28
...
Died on 28th day. Never showed trypano¬
somes. Tissues nil.
»
2 4
16
4
1
Trypanosomes always very scanty and some¬
times absent. Alive and well. ,
»>
26
10
6
Trypanosomes scanty. Accidentally killed on
10th day.
n
3 i
10
8
Trypanosomes very scanty. Alive and well.
»
3 2
6
...
Trypanosomes not yet seen. Alive and well.
»
34
6
...
Trypanosomes not yet seen. Alive and well.
35
6
Trypanosomes not yet seen. Alive and well.
Goat
11
34
...
Has never shown trypanosomes. Alive and
well.
»
12
34
...
Has never shown trypanosomes. Alive and
well.
Dog
*3
34
Has never shown trypanosomes. Alive and
well.
»»
*4
34
Has never shown trypanosomes. Alive and
well.
Monkey
21
22
10
Trypanosomes always scanty and sometimes
absent. Alive and well.
356
SUMMARY AND CONCLUSIONS
The trypanosome, of which a preliminary account is given in this
paper, seems to differ in several respects from a typical strain of
T. gambiense . In man it produces a form of sleeping sickness that is
relatively mild, occurs most commonly in young people, and in which
the trypanosomes are, apparently, either absent from the peripheral
blood altogether, or present in such small numbers that hitherto they
have not been detected. To the smaller laboratory animals the
strain seems to be but slightly pathogenic. The morphology of the
trypanosome as it appears in the blood of a guinea-pig shows some
peculiar features. The trypanosome is smaller than T. gambiense ,
the cell protoplasm when well stained is homogeneous, and there
appear constantly in the blood films a few very minute parasites
measuring as little as 8 fi in length. Some of the short and stumpy
parasites have the nucleus situated far forwards at the anterior
(flagellar) end of the body. The occurrence of a few peculiar
trypanosomes which appear to have a flagellum free in its whole
length is also remarkable.
Considering the morphological features of the parasite, and the
peculiar symptoms of the disease produced by it, I am convinced
that this trypanosome from Nigeria cannot be regarded as belonging
to the same species as T. gambiense. I therefore propose for it the
name T . nigeriense .
We regret that it has been found impossible to reproduce
Dr. Scott-Macfie's Plate in this number, but it will appear in the
next.— [Eds.]
REFERENCE
i. Stephens, J. W. W. and Fantham, H. B. Further Measurements of Trypancsoma
rbodes tense and 7 . gambiense. Annals of Tropical Medicine and Parasitology,
Vol. VII, No. i, 1913.
Volume VII
November, 1913
No. 3B.
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Edited by
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
AND
Professor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.,
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine,
The University,
Liverpool.
C. Tinting fr* Co. y Ltd.
Printers to the University Press of Liverpool
5 j Victoria Street
357
ADDENDUM ET CORRIGENDUM
The following Plate (PI. XXVIII) is of Trypanosoma nigeriense,
Macfie, 1913, and illustrates the preceding paper, already
published.
In the text of Dr. Mache’s paper the Plate was numbered
inadvertently as XXXIII. Hence, on pp. 344-346 the Plate should
have been referred to as PI. XXVIII.
35 «
EXPLANATION OF PLATE XXVIII
Trypanosoma nigeriense (Macfie, 1913)
Figs. 1 to 23. Forms of the trypanosome seen in the blood of a
guinea-pig. x 2,100.
Figs. 1-5. Long forms.
Figs. 6-9. Intermediate forms.
Figs. 10-13. Short, stumpy forms. In Fig. 13 the nucleus is seen
to be near the anterior (flagellar) end.
Figs. 16-19. Dividing forms.
Figs. 20-21. Trypanosomes containing granules and some
vacuoles.
Fig. 22. Rounded form.
TR TPANOSOMA N1GERIENSE
3 S 9
PRELIMINARY NOTE ON THE DEVELOP¬
MENT OF A HUMAN TRYPANOSOME
IN THE GUT OF STOMOXYS NIGRA
BY '
J. W. SCOTT MACFIE, M.A., M.B., Ch.B.
WEST AFRICAN MEDICAL STAFF
(Received for publication 14 September , 1913)
The role of Stomoxys in the transmission of trypanosomiasis
has been much discussed. The Sleeping Sickness Commission of
the Royal Society performed many experiments to determine
whether this insect could transmit T. pecorum. All the experiments
were unsuccessful, and Sir David Bruce and his collaborators con¬
cluded that ‘ It will therefore require very convincing proof to bring
this Commission to the belief that Stomoxys are carriers of this
disease.’ 1 A similar conclusion was arrived at by Bevan, 2 in
Southern Rhodesia, who failed to infect sheep with the Hartley
trypanosome by means of Stomoxys flies. On the other hand,
Bouet and Roubaud, 3 in French West Africa, succeeded in
transmitting T. cazalboui t T. pecaudi> T. sudanense, and T. evansi
by means of Stomoxys (probably S. calcitrans ). They did not,
however, observe any developmental forms in the digestive tract or
proboscis of the insect, and although they considered the species
of fundamental importance in the etiology of trypanosomiasis, they
were not able to regard it as a typical intermediary host. In
Northern Nigeria also this genus has fallen under suspicion, and
the writer has recorded one case in which an infection with T. brucei
( pecaudi) was probably conveyed to a horse by Stomoxys nigra or
Stomoxys calcitrans . 4
Last May, the writer had an opportunity of experimenting with
flies of this genus at the Medical Research Institute, Lagos. As he
succeeded in breeding Stomoxys nigra in captivity, it was hoped to
investigate thoroughly the role of this insect in the transmission of
trypanosomiasis. The experiments were however interrupted, and
360
as it seems unlikely that they can be resumed for some time, the
following incomplete records are given by way of a preliminary
note.
Towards the end of May, and at the beginning of June, 1913, a
number of Stomoxys nigra flies, caught in the laboratory, were fed
on a guinea-pig infected with the trypanosome from a case of
sleeping sickness from Eket in Southern Nigeria. The morphology
of this trypanosome, which is being described elsewhere, differed in
several respects from that of T. gambiense. Thirteen of the flies
were dissected from one to six days after the first infecting feed.
In six of them flagellates (Herpetomonas) were found in the mid¬
gut. As a control, twelve flies that had not fed on the infected
animal were dissected. No flagellates were found in them.
Unpcd Flies
Fed Flies
Number
Dissected
With
flagellates in
the gut
Without
flagellates in
the gut
Number
Dissected
With
flagellates in
the gut
Without
flagellates in
the gut
12
0
1
12 i
! 3
6
7
From these observations it was thought probable that the
presence of the flagellates in the mid-gut must be due to the
infecting feeds on the guinea-pig. As, however, it was possible that
the flies might have been naturally infected, experiments were
begun with flies hatched out in the laboratory.
Experiment 1. On June 14th a Stomoxys nigra fly that had
hatched out on the previous day was fed on the infected guinea-pig.
On June 17th the fly was killed and dissected. Herpetomonas were
found in the mid-gut. No flagellates were present in the salivary
glands and proboscis. The attached sketches, drawn with a camera
lucida, illustrate some of the flagellates found in the mid-gut of
this fly.
Experiment 2. On June 28th a Stomoxys nigra fly which had
hatched out the previous day was fed on the infected guinea-pig.
On July 2nd the fly died, and was dissected. Herpetomonas were
found in its mid-gut. No flagellates were seen in the salivary
glands and proboscis.
361
Each fly received only a single infecting feed. Two other
specimens of Stomoxys nigra that had been bred out in the
laboratory, but had not been fed on the infected guinea-pig, were
dissected. No flagellates were found in either of them.
Herpetomonas (X 1000) from gut of Stomoxys nigra.
At this stage the experiments had to be abandoned. Some
apology is necessary for recording such incomplete observations.
The two experiments given above were, however, quite definite, and
would seem to prove that the trypanosome with which the guinea-
pig was infected was capable of development in the gut of Stomoxys
nigra. I am not aware that this fact has previously been established,
and its importance must be my excuse for publishing this note. In
Nigeria, in the native towns and European stations, Stomoxys flies
abound. If, as appears probable, they are capable of serving as the
intermediary host of human trypanosomiasis, they deserve greater
consideration than they at present obtain.
Medical Research Institute,
Lagos,
Southern Nigeria,
August , 1913.
362
REFERENCES
1. Proc. Roy. Soc., 1910, B. 558, pp. 468-479.
2. Sleeping Sickness Bureau Bulletin, II, p. 252.
?. G. Bouet et E. Roubaud. Experiences de Transmission des Trypanosomiases animalw
de l’Afrique Occidentale fran^aisc par les Stomoxes. Bull. Soc. Path. Exot., 1912, v.
No. 7, pp. 544 - 550 .
4 Ann. Trop. Med. & Parasit., 1913, VII, No. 1, p. 9.
36.1
A NOTE ON A CASE OF LOA LOA’
BY
W. ROGERS, M.D.
(.Received for publication 30 August , 1913)
From May, 1906 to June, 1907, I was in Southern Nigeria;
beyond a few slight attacks of fever I was in good health. From
July to November, 1907, I was in Wales, and had evanescent
thickening of one or other leg, and, generally, an oedema about
the ankle, which puzzled me, and which lasted a day or two;
various diagnoses were made, including rheumatism, cellulitis,
erythema, nodosum, etc.
From November, 1907 to December, 1908, I was in Lagos
Protectorate; no symptoms beyond an occasional oedema about the
ankle of one or other foot; no fever. December—March, 1909,
travelling on Continent. In March, 1909, I settled down in Wales
and had no symptoms until the late summer, when I began to
notice fugitive swellings in various parts of the body, generally
in the neighbourhood of joints; painless, but associated with
stiffness. The general diagnosis was rheumatism, although one con¬
sultant of note diagnosed Angioneurotic oedema. In November,
1909, I recognised Filaria loa crossing the bridge of the nose, since
which time there has been no prolonged freedom from signs of the
presence of the worm.
As a rule, I became suddenly aware of a stiffness in a certain
part of the body; on examination I would find that part swollen,
sometimes to a considerable extent. At first the swelling was
wont to be localised, but later it would involve the greater part of
the circumference of the limb. I once had an upper arm that
Hackenschmidt would envy! When the parasite appeared on the
face I was generally made aware of its presence by accidentally
feeling a spot that was tender on pressure, say, in the lower jaw;
* The following note it of considerable value at representing Dr. Rogers' description of the
symptoms in his own case.—J. W. W. Stephens.
364
later on, in an hour or two, that part was swollen but the tender
spot had passed upwards towards the eye, always leaving in its
wake an oedematous track. I noticed that invariably when it
appeared on the face it made its way towards one or other eye: it
generally made one or two tours round the eye, either in the lid
or under the ocular conjunctiva, and then crossed the bridge of the
nose to the other eye; in the region of the eye it was always visible
under the skin. After visiting the eye it would wander over the
scalp, its curve being easily traceable by the tender area and, later,
localised swelling. The next day possibly, I would feel nothing
further, and so for several weeks; or, on the other hand, I might
wake up with a swelling in a leg or arm; in this case it is impossible
to say whether it was the same worm or no, as it was later shown
that I had at least two parasites. The presence of the worm in the
eye was always accompanied by a pricking, boring pain; when
under the conjunctiva it caused severe conjunctivitis with redness,
dilated vessels, feeling of grit, headache, etc.
On one occasion a cutting operation was attempted without
success; on another, an injection, likewise without success, of
1/1000 perchloride. After these attempts it disappeared for several
weeks. Towards the end of 1912 (about October, I think) a worm
was removed from under the conjunctiva, on which you kindly
reported, describing it as an immature female. After this there
were no more ordinary, usual symptoms for some time. In
December, 1912, I developed ‘Mumps,* and was accordingly
isolated; in the light of later knowledge I have no doubt this was
caused by the second parasite. After that I had rest until a month
or two ago, when, after the prolonged hot weather, the old signs
reappeared, but in an exaggerated form. A swelling would appear
in a limb, at first localised (exactly like a lipoma), but later on
(a few hours) firm, brawny and, for the first time, red , looking like
a cellulitis. The limb (arm or leg) would be quite elephantoid: on
one memorable occasion the thigh was so swollen that I was unable
for several hours to put on my trousers. On that occasion the
parasite , if it was the same one> had been seen in the eye the
previous day . The swelling would be totally gone in a day or two.
When it appeared in the eye it would leave that organ swollen and
closed, giving me a ‘drunk and disorderly* appearance; that, with
the accompanying headache and woebegone look, were likely to
give rise to unwarrantable suspicions detrimental to my character.
During the first two or three years I made numerous blood
examinations, but found nothing of interest. At first the red cells
were poor in shape and colour, and I several times saw the parasites
of tropical tertian fever. In 1909-1911 my mononuclear count was
about 20 per cent, and I had an eosinophilia of 3-5 per cent., which
was not surprising seeing that in addition to this F. loa I suffered
from Bilharzia (contracted during the South African War), and
from which I was not altogether free until the end of 1911. I have
made no blood examination since November, 1911.
A second worm, now in your possession, was removed in the
middle of August, 1913. (This worm is a mature female.
J. W. W. Stephens.)
367
RECENT EXPERIMENTAL RESEARCH
BEARING UPON BLACKWATER FEVER
BY
J. O. WAKELIN BARRATT, M.D., D.Sc. (Lond.)
{Received for publication 24 June, 1913)
In 1909 Barratt and Yorke* in an investigation into the
mechanism of production of blackwater showed that in this
condition haemoglobinuria was preceded by, and dependent upon,
haemoglobinaemia. It was also shown that suppression of urine,
occurring in blackwater fever, was due to mechanical blocking of
the uriniferous tubules. / In one experiment in which haemoglo¬
binaemia was induced in a rabbit by the injection of haemoglobin,
death occurred suddenly at the end of eighty-five minutes; in all
similar experiments, however, the general condition of the animals
injected was not affected .^
As subsequent papers bearing upon these observations are some¬
what scattered it will be of advantage to give a brief resume of the
results obtained.
Barrattf (1910), in order to ascertain the extent to which the
haemolytic action of quinine salts upon red blood cells was due to
hydrolytic dissociation, determined the first and second dissociation
constants of this alkaloid.
Barratt and Yorke* (1909) extended their observations to
piroplasmosis in the dog, and showed that in this condition
haemoglobinuria makes its appearance as soon as the blood plasma
contains as much haemoglobin as is obtainable from an amount of
red cells equal to 0*5 per cent., by volume, of the plasma, the
• J. O. W. Barratt and W. Yorke. ‘ An investigation into the Mechanism of Production
of Blackwater.* Annals of Trop. Med. & Parasit., 1909, Vol. Ill, p. 1.
tj. o. W. Barratt. * Uber die Konstanten der ersten und zweiten Dissociation des Chinins. *
Zeitschr. f. Elektrochemie, 1910, B. 16, S. 130.
t J. O. Wakelin Barratt and W. Yorke. * Uber den Mechanismus der Entstehung der
Hamofrlobinurie bei Infektionen mit Piroplasma cants.' Zeitschr. f. Immunitatsforschung u.
ezp. Thorapie, 1909, B. 4, S. 313.
368
percentage of haemoglobin in the urine being generally considerably
greater than that contained in the blood plasma. Haemoglo- •
binaemia and haemoglobinuria do not make their appearance in
this condition until an extensive destruction of red cells has taken
place, the latter often showing at the time of death a diminution
to one-fifth of the number originally present; the plasma, on the
other hand, remains little altered in volume. In two cases jaundice
was observed.
Yorke and Nauss* (1911) showed that the injection of a
haemoglobin holding fluid in sufficient amount was frequently
followed by severe symptoms, rapidly terminating in death. In
this connection attention may be directed to the severe symptoms
attending blackwater fever during the period of haemoglobinuria;
the similarity of the symptoms in the two cases suggests that the
mechanism of production is the same in both. Yorke and Nauss
also made the important observation that suppression of urine in
some cases followed the injection of a solution of haemoglobin.
It was found that this result was obtained if a sufficient amount
of the haemoglobin was injected in an animal which had previously
been deprived of water for some time, the occurrence of a greatly
increased secretion of urine, which would otherwise follow upon
injection, being thus prevented.
Yorket (1911) observed that, in haemoglobinaemia due to
Piroplasma cams or produced by the intravenous injection of
haemoglobin, granules reaching as much as 3 n in diameter were
found in large numbers in the epithelium of the convoluted tubules
of the kidney, indicating that the seat of elimination of haemoglobin
is the epithelium of these tubules. Illustrations are given of
sections of the kidney after injection.
Barratt and Yorke* (1912) in the course of a further investiga¬
tion showed that the acute symptoms following injection were not
attributable to haemoglobin, but to substances derived from the
stromata of the red cells. For the production of anuria the
• W. Yorke and R. W. Nauss. 4 The Mechanism of the Production of Suppression of Urine
in Blackwater Fever.’ Annals of Trop. Med. & Parasit., 1911, Vol. V, p. 287.
t W. Yorke. 4 The passage of Hamoglobin through the Kidneys.* Annals of Trop. Med*
and Parasit., 1911, Vol. V, p. 401.
t J. O. W. Barratt and W. Yorke. 4 Uebcr Haraoglobinamie.’ Zeitschr. f. Immunkatt-
forschung u. exp. Therapie, 1912, B. 12, S. 333.
369
introduction of haemoglobin in relatively large amounts into the
circulating blood was shown to be necessary, but here also the
presence of substances derived from the stromata was found to be
a factor in the determination of anuria. The blood generally
coagulated less rapidly after the injection of a solution of
haemoglobin. The observations made did not, however, admit of
a conclusion being arrived at as to the exact mode in which the
general symptoms following injection were brought about. In a
subsequent investigation* by the same observers these conclusions
were confirmed.
Barrattf (1913) pointed out that the granules in the convoluted
tubules of the kidney, observed in the course of haemoglobinaemia,
represent mitochondria, which in this condition become remarkably
enlarged, and are very readily stained. J
• J. O. W. Barratt and W. Yorks. * The Production of general symptoms in haemoglobin¬
aemia.*
t ]. O. W. Barratt. * Changes in Chondriosomes occurring in pathological conditions.'
Quart. Journ. of Micro. Sci., 1913, Vol. LVIII, p. 553.
X In this connection it may be pointed out that Lebedef! (‘ Zur kenntniss der feineren Verander-
ungen der Nieren bei Hamoglobinausscheidung.’ Virch. Arch., 1883, B. 91, S. 267) and Afanassiew
(‘ Uber die pathologisch-anatomisch Veranderungen in den Nieren und in der Leber bei einigen
mit Hamoglobinurie oder Ikterus verbundenen Vergiftungen.* Virch. Arch., 1884, B. 98, S. 460)
both give Ulustrations of the condition of the kidney in haemoglobinuria, in which the choqdrio-
somes of the convoluted tubules appear to be imperfectly represented. Marchand (* Uber die '
Intoxication durch chlorsaure Salze.’ Virch. Arch., 1879, B. 77, S. 455) regarded the elimination •
of red blood cells and portions of red blood ceUs as taking place through a land of diapedesis; the j,
structures in question being evidently hypertrophied mitochondria, which form striking objects in,
sections of the kidney during marked haemoglobinaemia. •
A FICTITIOUS NATIVE DISEASE
(ISIGWEBEDHLA)
BY
G. A. PARK ROSS, M.D., D.P.H.
GOVT. PATHOLOGIST BACTERIOLOGIST, NATAL
{Received for publication , 3 September y 1913)
For many years reports have been received from natives and from
Europeans resident in Zululand that a serious and often fatal
disease, known as Isigwebedhla, appeared regularly in epidemic
form about the same time as malaria. The outstanding feature of
this complaint was stated to be rapid ulceration of the rectum and
anus, described by some as gangrenous rectitis. The disease was
said to be characterised by sudden onset and fever. Within three
or four days the anus ‘ ulcerated/ in many cases to such an extent
that the closed fist might pass through it with ease, after which the
ulceration went on to attack the genitals, and even the mouth and
throat. As a rule there was a copious passage of green bile; some¬
times, however, mucus tinged with blood was passed, or even grey
sloughs. In very severe cases the disease ‘ attacked the back of the
neck.* Blood might appear in the urine, and in rare cases there
was haemorrhage from mouth and nose. The condition rarely
persisted for any time. In the majority of cases the patient either
died, or the disease, even when associated with the most extensive
ulceration, rapidly cleared up. This account of the condition was
corroborated by practically everyone I interviewed on the subject in
Zululand. It must be added, however, that few Europeans had
actually seen a case, and their impressions of it were derived from
native accounts. Only one post-mortem examination on a fatal case
had been made, and Dr. Manning, who performed it, said that he
could find no ulceration of the rectum whatever.
On arrival in Zululand I went somewhat further into the local
evidence, and summarise the more important particulars below.
37 *
(1) Isigwebedhla made its appearance shortly after the arrival
of the British in 1823, an event of political significance only.
(2) It occurs annually in epidemic form during and after the
rains.
(3) There is a great variation in the severity of the annual
epidemic, and in some years it is scarcely noticeable.
(4) This epidemic is always associated with a co-existent
epidemic of malaria (‘ Umkuhlane ’).
( 5 ) Isigwebedhla occurs only in the bush veldt among the Zulu
people, unlike malaria which is endemic on the coast flat among the
Tonga and mixed race found there.
(6) It has no relation to the tsetse belts, or to the parts infested
with the tick Ornithodorus moubata which carries the parasite of
relapsing fever.
(7) Dietetic changes, as the use of green mealies, new mabele
(millet grain), etc., have no influence on its appearance. The new
crop of cereals begins to be used at the time of its greatest incidence.
(8) One attack does not protect against a second, and a person
may suffer two, or even more, times from it in the course of a single
year.
Accompanied by a posse of native doctors, I watched the
progress of nineteen cases, each and all of which were diagnosed by
these persons as typical and severe ‘ Isigwebedhlas.* In eighteen of
these I demonstrated either in the peripheral, splenic, or hepatic
blood the presence of Haemamoeba praecox of malignant tertian
malaria. In some instances it was in prodigious numbers. All my
cases showed the other cardinal signs of malaria, e.g., pigmented
leucocytes and a mononuclear increase, and all recovered rapidly
under quinine injections. The clinical types noted were as under:
14 were typical bilious remittents.
2 were of the dangerous algid type with great cardiac weakness
and a tendency to collapse.
3 were cerebral in type, 2 showed coma, and the other was
characterised by convulsions, delirium and diarrhoea.
True dysentery was not a feature of any of them, although in
one case the motions contained blood, but only for a day.
In all, the malarial intoxication was sufficient to markedly lower
the tonicity of the sphincter ani, and permit of its easy dilation.
373
In two cases stated to be exceptionally severe, slight rectal prolapse
was present. Both patients were more or less collapsed, and in one
the condition was undoubtedly aggravated by a retroverted uterus
and two days of irritating diarrhoea.
In all, the rectal mucosa participated in changes similar to those
observed in the buccal mucosa in any case of fever. Trifling
excoriation was observed in most, and was invariably described by
those present as ulceration. No organisms were ever found in the
tissues below the * ulcerations/ and scrapings taken from the parts
the natives considered ulcerated showed only the ordinary organisms
of the intestinal tract. In the case noted above, in which convul¬
sions were a feature, the diarrhoea was apparently due tp a small
spirochaete, identical as to staining reactions, and presenting the
same relations with the endothelial cells of the gut, as that described
by Le Dantec. Certain ‘ abdominal 9 features of the case may have
been due to this organism, but I do not consider that its importance
in this instance as a disease-producing factor compares with the
r 61 e played by the sub-tertian malaria parasite, with which even the
peripheral blood was heavily infected. This spirochaete is said to
produce a mild diarrhoea, chronic in type, in the South of France,
and it is possible it may do the same in Africa. It can be easily
overlooked on account of its small size and the difficulty of staining
it. Those found by me measure 5 to 10 p by 0*5in width.
They stain faintly pink by Leishman, and are best shown by
weak carbol-fuchsin. They do not take Gram. They are seen
as minute coiled threads little longer than a red blood corpuscle.
They may occur almost in pure culture, forming in places dense
felted networks. A few are seen clinging to the free surfaces of
the endothelial cells. They were absent from scrapings taken from
the rectal mucosa, and must have had their origin higher up. None
of my cases showed Sp. vincenti , nor organisms which could be
identified with the Bacillus fusiformis.
Herpes on the lips, buttocks and perineum was occasionally
observed. This is a common concomitant of malaria, and explains
the reports of 4 ulceration involving the skin/
In the mouth sores, and those involving the throat, in the only
case in which the latter was ulcerated, nothing of interest was seen.
374
Sp. dentium was once demonstrated, and in two instances I found
extensive infection with Sp. buccalis.
The absence of eosinophilia in all precluded the likelihood of
serious hook worm infection. I searched the dejecta in each case,
and failed to find eggs or embryos. Ascarides and tapeworm were
common enough. I saw no cases of rectal bilharzia. Routine
examination of hepatic blood failed to disclose Leishman-Donovan
bodies, and none of these cases showed trypanosomes or Sp. duttoni ,
a matter of some interest when compared to the state of affairs in
the low veldt proper, where the existence of Isigwebedhla is denied.
It becomes necessary, however, to explain the apparent
discrepancy between native accounts of the disease and the actual
facts as determined by my investigation.
The initial treatment of most acute diseases, and specially of
specific fevers among the Zulu, consist in taking an enemetic
followed by an enema. This is done to remove ‘ bile. 1
Emesis is frequently produced by drinking plain water, but in
the majority of instances a host of bulbs, roots and barks pounded
up, and used either singly or in combination, are infused in hot
water which is then drunk. Most of these substances are harmless,
many of them, as the Mfusamvu (Pittosporum viridiflorum ), act in
virtue of their soapy nature, a few are true emetics, and some, as
the Macapazane (Bowiea volubilis ), the juice of which sets up a
papular rash on the skin and proves toxic to guinea-pigs in small
doses, are irritants.
The rectum is emptied as follows: The patient is placed in the
knee-elbow position with the chest resting on the ground. A cow-
horn with the point cut off is inserted into the anus and a watery
infusion of an extensive series of roots, bulbs, etc., is poured in
until the bowel will hold no more, when the enema is evacuated.
This process is repeated until the fluid comes away clear, and is
continued daily. The enema is often followed by a suppository,
or the parts are painted with a decoction of various bulbs. I have
been told that in Isigwebedhla a suppository of mud and red chillies
is sometimes used, ‘ with excellent results.*
After one or more days of severe malarial fever the tone of the
sphincter ani is so reduced that traction on the buttock of a patient
in the knee-elbow position results in an immediate dilatation of the
375
anus. The degree of patency depends on the success with which one
can overcome the resistance of the sphincter, and, other things being
equal, varies directly with the gravity of the constitutional effects
of the fever. The patency, which in many of my cases was extreme,
is to the native mind the * eating away of the flesh/ i.e., ulceration,
and is the one cardinal feature of the ‘ disease/
It is customary in these cases to keep the patient sitting up
throughout the course of the sickness, to facilitate the easy flow of
bile. This is done by relays of friends, and it is not surprising to
find the unfortunate patient unable to hold up his head after two
or three days. On complaining of pain in the spine or back of the
neck, the diagnosis that ‘ Isigwebedhla * has affected these parts is
quickly arrived at.
Malaria is, if anything, more prevalent among the coast people
than among the Zulu of the hills. There are places in Hlabisa
district where one sees cases of 1 Isigwebedhla 1 in the Zulu kraals
among the hills, and hears of dozens in the immediate neighbour¬
hood. Not two miles away on the flat are people who are
suffering from malaria in as great a degree, yet deny the existence
of * Isigwebedhla' among themselves, and invariably affirm that it
is a condition confined to the Zulu.
I consider that the term ‘ Isigwebedhla * is an invention of some
astute practitioner in the past, and have had some strong hints as to
the accuracy of this view of it given me by Tonga 1 doctors' whose
clientele embraces Zulus. Rightly or wrongly, the Tonga doctor
has a greater reputation than his Zulu confrere, and it is not only
possible, but likely, that this myth was evolved by the Tonga.
There is no doubt that in judging of the severity of such a case
the native is swayed more by his deductions from the constitutional
symptoms than by the presence of definite lesions in the rectal
mucosa, and he is much influenced by the rapidity with which the
constitutional symptoms develop. At the same time he is unable to
imagine grave constitutional symptoms existing independently of a
gross lesion to account for their production. Given delirium or
coma (unless he can explain it by witchcraft) he readily constructs
an alarming pathology to account for its existence. A comatose
case is ‘ dead 9 of Isigwebedhla; he may take a hasty glance at the
rectum. If no ‘ulceration' is seen, or the patency is not marked,
376
the former must certainly exist higher up. He would never ascribe
symptoms to a blood condition per se , even if he could conquer his
intuition to ascribe such serious disease to witchcraft, an idea which
is strengthened by the absence of gross morbid changes.
Seeing, therefore, that ‘ Isigwebedhla * is merely an advanced
stage of an acute febrile disorder, it is difficult to refrain from
admiring the acumen by which these ignorant (sic) practitioners,
recognising their utter inability to effect a cure, have evolved this
terrible and fatal malady, the successful treatment of which is
unexpected at best, and which, if achieved, cannot fail to reflect
the highest credit on all concerned in it.
377
ON ‘VOMITING SICKNESS’ IN JAMAICA
(Report of the XXXth Expedition of the Liverpool School of
Tropical Medicine)
BY
HARALD SEIDELIN, M.D.
(Received for publication 25 June, 1913)
PLATES XXIX-XXXIII
In December, 1912, the Committee of the Liverpool School of
Tropical Medicine resolved to send me to Jamaica in order to
investigate the nature of the disease called ‘ vomiting sickness/
prevalent in that island during the winter months, and responsible
for a considerable mortality, chiefly amongst native children. It
was considered of particular importance, for obvious reasons, to
determine whether a relationship exists between ‘vomiting sickness’
and yellow fever.
The Committee of the School received, through the Colonial
Office, the assurance that the Government of Jamaica would do all
in its power to further the objects of the expedition.
I left Southampton on December 18th, on the R.M.S.P.
‘ Thames/ and arrived in Kingston, Jamaica, on January 10th, 1913.
The Government, through the Superintending Medical Officer,
Dr. J. Errington Ker, at once made suitable arrangements in order
to facilitate my task. It was agreed that I should establish my
headquarters in Kingston, where I could carry on laboratory
investigations, and at the same time be in easy communication with
all parts of the island, always being ready to go out into the country
at a moment’s notice. Instructions were issued to all the District
Medical Officers, requesting them to report by telegraph all cases of
' vomiting sickness 9 which might come to their notice, and to make
arrangements for post-mortem examination if the cases ended
fatally before my arrival. These instructions were faithfully
carried out, and on January 13th I performed my first post-mortem
examination at May Pen. The only drawback, which was some-
378
times seriously felt, was that of the very considerable distances which
I had to travel in order to reach a case. Cases often occurred out
in the bush; in many of these cases the authorities took the trouble
of transporting the patient or body to the nearest hospital or
mortuary; but this was not always possible, and sometimes it was
necessary to travel first several hours by rail, and then several hours
in a buggy, besides walking the last part of the way. Two
inconveniences resulted: that it was impossible to see even
approximately all the cases occurring and reported; and, secondly,
that I usually arrived late, in fact, as a rule, after the patient had
died, and in some cases even somewhat late for a post-mortem
examination. The transport of material to Kingston, for patho¬
logical and bacteriological examination, was also sometimes delayed
more than was desirable.
Under these conditions I was unable to see more than about half
the number of cases reported during my stay in Kingston. One
part of the island, the parish of Trelawny, I was particularly
anxious to visit, as the majority of the cases diagnosed by Captain
Potter as yellow fever had been observed there. I had, however,
no opportunity of doing so; very few cases occurred this year in
Trelawny, and these I could, for various reasons, not reach in time.
Otherwise I saw cases, sometimes alive, but more often after death
only, in nearly all the various localities where the disease occurred,
and my material includes, as far as I can judge, all the divers
types which, with any degree of reasonableness, can be included in
discussions on ‘vomiting sickness.’ Altogether, I saw sixty-two
cases (besides several which could at once be otherwise diagnosed),
a fairly large number, considering that my whole stay in Jamaica
lasted only ten weeks. In the meantime I carried on laboratory
work in the Laboratory of the Public Hospital, Kingston; here
I had the invaluable assistance of the Government Bacteriologist,
Dr. H. H. Scott, who received the material which I sent in from
the country, incubated and examined many of the cultures, and
always helped me in every possible way. Without his kind
assistance I should have been obliged either to refuse a large number
of calls or to leave the larger part of the laboratory work undone.
The described conditions of work in Jamaica made it impossible
to investigate all epidemiological, clinical, anatomical and micro-
379
biological details in each case as fully as might have been desirable.
The histological work was only just begun in Jamaica, but
was almost wholly carried out in Liverpool, and so far
under better conditions, as no fresh cases of ‘ vomiting sickness *
were continually claiming the attention, of the worker. But various
reasons made it desirable not to devote too much time to this part
of the investigation. Other work was waiting at home and it was
desired that I should as soon as possible proceed to Africa on
important investigations. Under the circumstances the necessary
examination of sections was done to establish an anatomical
diagnosis in doubtful cases and confirm it in others, and illustrations
were prepared in order to show the more important histological
changes, but no attempt was made to describe the microscopical
appearances in all details. This would have been a work of years,
whilst at present only about two months could be devoted to it.
Instead of delaying the publication of the report it was therefore
thought advisable to publish it as soon as possible, especially
because there seemed to be no probability that a closer study of
histological details, though in itself extremely interesting, would
throw any further light on the one question of paramount
importance, the question of etiology. I must, however, reserve for
myself the right of publishing later on more detailed descriptions
of some of the cases which present certain features of more than
ordinary interest, though, I believe, without any bearing upon the
question of the nature of ‘ vomiting sickness.’
On March 22nd I sailed from Kingston on the R.M.S.P.
* Oruba/ and on April 15th I arrived at Southampton.
The report is divided into the following sections : —
I. Historical notes (p. 380).
II. Recent observations (p. 386).
III. Personal observations (p. 390).
IV. Discussion of personal observations (p. 393).
V. Description of vomiting sickness (p. 450).
VI. General discussion of its nature (p. 454).
VII. Prophylactic measures (p. 466).
VIII. Other observations in Jamaica (p. 466).
IX. Other places visited (p. 467).
X. Acknowledgments (p. 468).
380
I. HISTORICAL NOTES
According to Potter (1912) the term ‘vomiting sickness* has
apparently been in use in Jamaica for many years. My own
impression was the same, but I could obtain no definite information.
Being a disease which chiefly attacks native infants and children,
it seems quite likely that it has not attracted the attention of
medical men to any considerable degree, as infantile mortality has
been and still is very high, especially among the natives. Native
children are often ill, and die, without medical attendance, and in
cases of ‘ vomiting sickness * it must be admitted that parents have
one particular and often very good reason for not applying for
medical aid, namely the extreme acuteness of the cases which, in
combination with the long distances in many cases, makes it useless
to send for a doctor as the child is usually dead before he arrives.
In the medical literature 1 vomiting sickness * is apparently
mentioned for the first time by Turton (1904), who gives a general
description of the disease, summing up in the following way: —
‘ We have here a disease which occurs only in the cold months,
in country districts, among the poorer classes and unhealthy
surroundings; may attack more than one member of the family,
either at the same time or at intervals of hours up to a day or two;
usually attacking children irrespective of sex; presenting a longer
or shorter period of malaise , followed by vomiting, convulsions,
and death in a few hours; showing, post mortem , signs of gastro¬
intestinal irritation, and usually infection, with ascarides in large
numbers; killing by respiratory failure; and having a very small
death rate in cases where early treatment has been given.’
Turton is of opinion that the post-mortem appearances
are due to 1 venous congestion,* and has, in fact, constantly
found hyperaemia of the following organs: cerebral and spinal
meninges, lungs, liver, spleen, kidneys, and stomach. Some
importance is apparently given to the presence of Ascarides, which
were found in nearly every case in the small intestines, sometimes in
enormous numbers and forming knots, causing distension of the
bowel and irritation of its wall.
In earlier years the same disease may have figured in the Health
Reports under other headings, especially under that of cerebro-
3 »i
spinal meningitis; this question is discussed by Potter (1912,
Appendix A). In 1903 the term ‘vomiting sickness’ appears as a
synonym of gastro-enteritis (Annual Report of the Superintending
Medical Officer), and in the following years the disease is
referred to as ‘ vomiting sickness.*
In 1905 the Superintending Medical Officer, Dr. J. E. Ker,
draws attention to the prevalence of ‘ vomiting sickness * and to the
uncertainty which reigns with regard to its nature; the great
interest and importance of the matter are pointed out, and the
following points are stated as certain: —
1 1. The disease appears at a certain fixed time of the year,
November to March, at a time when the temperature varies greatly
from day to night.
‘ 2. It rarely appears in town; none of the cases reported have
come from towns.
‘ 3. The people attacked are chiefly, but not always, children.
‘ 4. It appears so suddenly and runs its course so quickly that
medical men never hear of two-thirds of the cases until after death
has occurred.
‘5. Frequency with which several members of a family are
attadced.*
Ker states that the affection has been returned under various
names on the death certificates, and quotes 1 Cerebro Spinal
Meningitis, Gastritis, Cerebro Spinal Fever, Vomiting and Fits,
Black Vomit, Vomiting and Pain, etc.*
After this introduction Ker publishes a number of reports on
‘ vomiting sickness * by several District Medical Officers who were in
possession of considerable experience with such cases.
Dr. Cooke, Mandeville, writes, ‘ There is no such thing as
" Vomiting sickness.** The vomiting is simply a symptom as it is
of many other diseases. The disease is acute congestion of the lungs,
occurring in children under 5 years of age, and only in the poor,
ill-fed, and almost naked.* The description of cases given by
Cooke appears to bear out his contention.
Dr. Thomson, Chapelton, describes an outbreak of ‘ vomiting
sickness which occurred after a period of unusually cold weather.
‘ Cases occurred at the same time in widely separated portions of
the district. The disease has been more virulent than in previous
382
years, more adults have been attacked than in previous epidemics,
and the death rate has been enormous. So rapid is the course of
the disease, that in many cases there has been no time to obtain
assistance. Very few cases of the disease are seen, only those
occurring in the neighbourhood of the village are reported, the
others being heard of only after death has occurred, being reported
as sudden death to the police.' Two typical examples of the
history are given as follows:—‘ I. A young child of healthy
appearance is put to bed. The parents are roused during the night
by a cry and find the child foaming at the mouth, retching,
unconscious and cold, with twitching of the limbs, which passes into
actual convulsions. 2. A young adult, more often female, leaves
home for market—is attacked on the way with retching, vomiting,
and intense weakness, becomes unconscious, and is brought home a
corpse. Post-mortem examination of these cases reveals no naked-
eye appearances of disease. The mucous membrane of the stomach
may or may not be congested from the vomiting. There is never
any trace of an ulcer. No irritant poison has ever been detected
in the cases submitted for analysis. There are never any signs of
disease in the kidneys or meninges.'
Dr. Earle, May Pen, describes the disease as apparently ‘a
severe form of gastritis, with rapid collapse and shock,' and states
that it is ‘ very fatal, especially amongst children.'
A report by Dr. Bell, dated 1900, deals with the disease as
observed in the same district, May Pen. He gives detailed
descriptions of both clinical symptoms and post-mortem findings.
Malaise with epigastric pain is followed by mucous vomiting, and
prostration. After an interval of apparent improvement the
vomiting returns and collapse sets in. Death may occur after a
more or less rapid course. In some cases children were found
during night with convulsions, which soon ended in death, or
vomiting set in suddenly, with collapse, and death ensued in a few
hours. At the autopsies hyperaemia of meninges, brain and liver
was observed; the spleen was enlarged, but only slightly congested;
the kidneys were intensely hyperaemic, and the stomach showed
ecchymoses in the mucosa. The mortality was only about 2 per
cent, if treatment was instituted early; otherwise, treatment was of
no avail. The term 'epidemic gastritis' is not considered
3»3
satisfactory, and no definite opinion is stated with regard to the
nature of the disease.
Dr. Tillman, Vere, states that it was ‘ almost entirely a question
of food and clothing, as out of over nine hundred cases treated,
not one case occurred among the coolie-children of the district/
Tillman diagnosed the first case which he observed as one of
meningitis, but later on he arrived at the conclusion that the
meningeal hyperaemia was a secondary phenomenon, due to lung
stasis and stasis in the venous system; the gastro-intestinal
hyperaemia and irritation are considered phenomena of primary
importance. The cause of the whole disorder was thought to be
atmospheric. He reports on 144 cases observed, with only two
deaths. He describes general malaise and sudden severe pain in
epigastrium with violent vomiting, anxious expression, weak, rapid
pulse, and slow and shallow respiration. The temperature is said
to be often normal, but in most cases rising to 102 or 103, generally
preceded by a rigor. Convulsions and death follow, unless
treatment is resorted to, in which case the convulsions may be
represented only by contractions of the feet, and develop no further.
The anatomical lesions found by Tillman were ‘erythematous
appearance 9 of the gastric mucosa, hyperaemia of liver, spleen,
kidneys, and meninges, and hyperaemia and oedema of lungs.
The paper by Dr. Turton, Stony Hill, which is mentioned above,
is also included in this report.
Dr. Calder, Santa Cruz, states that he has seen no cases exactly
corresponding to the type of ‘ vomiting sickness * as described by
his colleagues, but reports two cases in which vomiting was a salient
feature, and in which he has performed post-mortem examinations.
One was a child, the other an adult; pancreatitis, jejunal hyperaemia
and haemorrhage, and petechiae in the stomach were observed in
both cases.
It is obvious that various diseases may have been referred to in
these reports. One difference is particularly striking between the
views expressed in the reports of Bell, Tillman, and Turton, and
those held by the other District Medical Officers. According to the
latter, ‘vomiting sickness’ is an extremely fatal disease, whilst the
type described by the latter is fairly benign, the mortality being
as low as 2 per cent., or even less. It might be supposed that only
384
the latter had had the opportunity of seeing cases m the early
stages, whilst the Medical Officers of the former group had always
been too late, because of longer distances or for other reasons. As
no specific treatment is indicated, it appears, however, entirely
unintelligible that the character of the disease should have been so
completely transformed by the measures described. Two
possibilities remain, either that some observers have overlooked mild
forms of the disease, or that others have included cases of a different
nature. Neither possibility can be dismissed, but the latter seems
perhaps the more probable. Many of the recovering cases would
evidently, in the usual course of events, have been diagnosed as
acute gastritis or gastro-enteritis, especially those with febrile
temperatures and without collapse.
Dr. Ker concludes his report with the recommendation that
special and systematic investigation into the question of 1 vomiting
sickness ’ should be made at an early date.
So far, in 1905, the entity of 1 vomiting sickness * has not become
established, a fact which must be borne in mind when discussing
the following reports.
The Annual Report (1906) contains brief notes about ‘vomiting
sickness/ from various districts; it appears that the disease had
been less prevalent and perhaps less malignant than in previous
years. Two post-mortem reports, by Dr. Earle, are published; in
both cases there was hyperaemia of meninges, lungs, liver, spleen,
and kidneys. Petechial spots were observed in the brain substance,
and subepithelial ecchymoses in the stomach. The descriptions are
almost identical in the two cases, which are considered typical
examples of the so-called ‘ vomiting sickness.*
In the 1907 Report it is stated that ‘ vomiting sickness * has been
more prevalent and more severe than usual. In Clarendon some
80 or 90 deaths were actually reported, but many more are believed
to have occurred. In Trelawny both adults and children were
attacked, with fatal results. The need of a thorough investigation
is again pointed out.
In 1908 ‘ vomiting sickness * is said to have been little prevalent.
In 1909 the disease is reported present in Falmouth and
Chapelton, but has evidently not been very prevalent.
385
In the 1910 Report exceptionally severe outbreaks are reported,
and the following interesting table is given: —
'Number of deaths recorded as having been caused by
Vomiting, Vomiting and Fever, Vomiting and Convulsions,
etc.
Parish
Month
Total
Dec.,
1908
Jan.,
1909
Feb.,
x 9°9
Mar.,
I 9°9
Dec.,
x 9°9
1910
Feb.,
1910
St. Andrew .
6
,
■j
-
3
8
16
35
St Catherine
9
4
1
22
10
1
5 1
Trelawny .
*5
38
1
7
12
9
IOI
Westmoreland
2
—
1 I
1
3
5
—
H
Clarendon .
14
9 1
43
*9
230
St Thomas .
—
1
KB
4
8
1
x 4
Hanover.
3
7
8
I
2
1
3
2 5
St James .
4
20
■HI
10
3
4
57
Manchester .
7
13
3
47
x 9
1
93
620
In the same Report the following statement is made by Dr. Ker :
'That many of these cases may be, and undoubtedly are, due to
ackee poisoning, possibly cassava poisoning, worms, Meningitis,
Bronchitis, Gastritis, and other diseases, there is little doubt, but
beyond these there remains an uenxplained residue which needs
explanation/
In this connection the frequency of helminthiasis in children is
mentioned; one District Medical Officer found, on administration
of vermifuges, that out of 72 apparently healthy children only two
expulsed no worms, whilst two passed more than a hundred ascarides
each.
Cases and deaths are reported, in 1910, from many districts—
it is evident that more attention is now being paid to the disease.
Dr. Tillman reports 1,200 cases, the mortality not being stated.
3 86
Tillman modifies his earlier statement about coolie children: they
do not escape altogether, but practically so; the different suscep¬
tibility is explained by the coolie children being better cared for
than the children of the natives.
In the 1911 Report Dr. Ker abstains from discussion of the
subject, awaiting the result of Captain Potter’s investigations
carried out during the corresponding financial year. Outbreaks of
4 vomiting sickness ’ are reported from Stony Hill, Claremont,
(St. Ann), St. Ann’s Bay, Duncans (severe), Falmouth, Montego
Bay, Newport, Crofts Hill, and Chapelton; in the report from
Ulster Spring it is stated to have been ‘ practically absent.’
In 1912 no special reference to ‘vomiting sickness’ is made in
the Annual Report, but two cases are stated to have died in Public
Hospitals, the locality not being given.
II. RECENT OBSERVATIONS
It is obvious, from the data given in Section I, that a great many
factors have been submitted for discussion as probable causes of
4 vomiting sickness.* The existence of a separate morbid entity has
been denied by several observers, but as a rule each observer has
given his own view with regard to its nature, and these views have
differed considerably—from helminthiasis to bronchitis, and from
ackee poisoning to meningitis. The prevalent idea, however, has
probably been that which was pronounced by Ker, that a group of
unexplained cases remained when those were separated out which
could be classified as well-known diseases. During the last year
or so the discussion has taken on a somewhat more definite
character.
Potter (1912), in a Colonial Office Report, published the first
systematic investigation into the nature of ‘ vomiting sickness,’
making the startling announcement that the majority of the fatal
cases were yellow fever. This had probably been suspected by
many, outside Jamaica, considering that the name of black vomit
is given as a synonym of ‘ vomiting sickness ’ in Ker’s summary,
and remembering the existence in other West Indian Islands of the
so-called ‘ fievre a Vomissements noirs des enfants,’ by several
authorities regarded as a form of yellow fever.
3«7
Potter does not declare that 1 vomiting sickness’ is identical with
yellow fever; he agrees with other observers that the former name
has been applied to a great many different diseases, and his report
contains abundant evidence that this is the case. A large number
of cases are included which have evidently nothing whatever to do
with the type under discussion. Numerous cases are malarial, and
others are otherwise definitely diagnosed. One group of seven
cases, described in Appendix D to Potters Report, shows very
clearly the abuse, even of a most dangerous nature, to which an
ill-defined name may lend itself. Five fatal and two non-fatal
cases occurred in one hut, the District Medical Officer at first
diagnosing 1 vomiting sickness/ but afterwards suspicion of
poisoning arose. One of the bodies was exhumed and arsenic was
found in the organs. In spite of this evidence the investigation
was not pursued, and the jury found that the cause of death was
‘vomiting sickness.’
Eliminating from the discussion all these cases in which some
other definite diagnosis can be made, Potter is of opinion that the
remaining cases show the clinical characters and post-mortem
appearances of yellow fever.
Before considering the question of yellow fever, Potter discusses
the explanations previously advanced, and arrives at the conclusion
that they are all unsatisfactory. With regard to cerebro-spinal
meningitis he regards it as impossible that a large number of
fulminant cases should develop without the simultaneous occurrence
of other cases running a more prolonged course; besides, he says,
in cases which had recovered, * sequelae, such as paralysis, blind¬
ness and deafness should be expected/ and in fatal cases ‘ the post¬
mortem examinations would have shown definite inflammatory
changes in the meninges.’ The hypothesis of ptomaine poisoning
has not been supported by symptomatological, nor by circumstantial
evidence. The question of ackee poisoning as a cause of death—
in some cases—is left open. Cassava poisoning is considered out
of the question, as in that case the presence of prussic acid, the
poisonous agent, would probably have been noticed.
Potter observed personally 73 cases, clinically or post mortem
(the numbers run only to 70, but two different cases are both
numbered 43, and the two cases in the additional section of
388
Appendix B were also seen by Potter); in addition he has collected
a considerable number of observations from official reports. In
many of the cases, both personal and from other sources, a definite
diagnosis of malaria or some other affection was made, but a
considerable number remains in which Potter suspects yellow fever,
and in seventeen cases he feels certain of this diagnosis. In these
seventeen cases the character of the stomach contents and the lesions
of the gastric mucosa are certainly often suggestive of yellow fever,
but the changes observed in other organs are usually so
incompletely observed that the reader cannot possibly appreciate
their nature and significance. It is very remarkable, and by no
means in accordance with common experience in yellow fever, that
the kidneys are often stated to be normal, and the urine free from
albumin. The same objection which Potter makes against the
assumption of the cases being cerebrospinal meningitis can, with
equal right, be made against a diagnosis of yellow fever: one
would certainly expect that other cases occurred at the same time
with more typical symptoms and a more prolonged course. To
the reader of the report it is a great inconvenience that detailed
descriptions are very seldom given, and that the findings recorded
are not fully considered in the light of yellow fever pathology and
epidemiology.
Potter’s conclusions were at once contradicted by the Governor
of Jamaica, in a letter to the Colonial Office. In support of his
letter the Governor publishes a report by Scott (1912, 1), containing
observations on cases of ‘ vomiting sickness,’ with demonstration of
meningococci in the spinal fluid. Later on, Scott recorded further
observations in two Official Reports (i9 I2 » 2 an d 191), and soon
afterwards published a more detailed paper on the same subject
(I 9 I3 > 2).
In this last paper Scott summarizes his results. His first
observations are on cases of meningitis of an ordinary type in which
he found meningococci. Subsequently, he observed some cases
of a fulminant type, showing the characters of ‘ vomiting sickness,’
but occurring at a time of the year when this disease was usually
not prevalent. All cases—five—occurred in one family. Inves¬
tigation for meningococci was made in four out of the five cases,
and the organism was found each time. In addition, meningococci
389
were found in the rhino-pharynx of a boy in the same family where
the cases had occurred. Scott now followed up this line of
investigation, and soon succeeded in finding the same organism in
a considerable number of cases. These were all rapidly developing,
but not all fatal; a considerable number belonged, undoubtedly,
to the type of ‘vomiting sickness.* Scott’s paper deals with
52 cases, of which only a comparatively small number are personal
observations, except for the examination of the spinal fluid, which
he performed in 33 cases, the fluid either being obtained by himself
or sent in by the Medical Officer in charge of the case. Meningo¬
cocci were found in 23 cases, and similar cocci which could not be
satisfactorily identified, in a few instances. In two cases
spirochaete-like bodies were observed in the spinal fluid; both
cases were of a rapidly developing type, and in one, at least, were
meningococci also found. The spirochaete-like bodies were only
seen on direct examination, but did not develop in cultures. The
author leaves the question of their nature and importance open.
No anatomical lesions of any importance are mentioned in
Scott’s paper, except in the case of the meninges, and even in this
case very few details are given. Little attention has evidently
been paid to the pathology of the disease, which may, to a large
extent, be accounted for by the fact that so few cases are personal
observations.
Scott prepared vaccines from some of his cultures from the
earlier cases, and used them later on in several mild cases and in a
number of contacts—apparently with success. He fully realizes
that vaccine treatment can have but little chance of success in the
hyperacute cases, and, whilst recognizing the possible value of
serum treatment, at the same time points out that its application
meets with so many practical difficulties that it is impracticable in
the majority of cases.
Scott’s work furnishes a very definite basis for discussion. Since
the appearance of his paper no further investigations have been
published on the subject, but MacDonald (1913) has discussed and
criticized Potter’s paper, pointing out that he had failed to take up
several important lines of investigation, and that no conclusive
evidence had been brought forward in support of the diagnoses
of yellow fever.
390
III. PERSONAL OBSERVATIONS
During my stay in Jamaica I observed 62 cases which were
presented to me as cases of 4 vomiting sickness/ and in which no
other definite diagnosis could be made at once. A closer
examination, however, revealed considerable differences, which make
it necessary to divide the total number of cases into several groups
for separate discussion. A brief summary of all the cases is given
in the accompanying Table I; it has not been possible to include all
symptoms and anatomical lesions, but, as it stands, I believe it
conveys a fairly correct, general impression of the character of
disease with which we have to deal in 4 vomiting sickness/
Detailed histories of some of the cases—as far as they were
obtainable—will be given further below when the various groups
are discussed.
In addition to these personally observed and ‘suspicious-
looking * cases, I received from various colleagues observations
which have not been included in the table, but some of them will be
referred to later on. Nor have I included personal observations
in which the symptoms at once suggested some other diagnosis than
that of 4 vomiting sickness/ unless the further course of events again
appeared to bring this diagnosis in the foreground.
A few explanatory remarks are necessary in order to define
more clearly the too brief headings of some of the columns in the
table, and some indication of the pathological and bacteriological
technique employed is also due.
First a general remark: the sign + indicates that the corre¬
sponding symptom was present, the sign — , that it is stated to
have been absent. No sign means that no definite statement has
been made in my notes; in this case, however, the symptom has
usually been absent.
With regard to the clinical symptoms, it must be remembered
that the majority of the patients did not come under medical
observation during life. We therefore had to rely in these cases
on layman’s evidence, from which it, as a rule, was possible to
elicit the presence or absence of symptoms as vomiting, convulsions,
and coma. We found also that the statements with regard to the
characters of the vomits usually appeared reliable. The vomited
Note. — 'Cable 1 giving a synopsis of the cates will be found opposite p. 468.
matter was described, in the vast majority of cases, as watery or
mucous, and sometimes it was said to have been greenish. With
regard to black vomit, it was only described twice, and in two
instances has its presence been queried. In one of the latter cases
it was suspected because black stomach contents were found at the
post-mortem examination, but nothing definite could be ascertained
with regard to the appearance of the vomited matter. In the other
case the grandmother of the patient, when telling the story of the
child’s illness, which had upset her considerably, to begin with
mentioned black vomit, but afterwards denied emphatically to
have said so, stating that the vomits had been absolutely colourless.
The column * emaciation ’ has been inserted on account of the
often-expressed opinion that ‘ vomiting sickness ’ is a disease chiefly
or exclusively of emaciated or poorly nourished children. This
opinion is emphatically contradicted by my experience, as the
answers show. The sign + in this column does not exactly mean
extreme emaciation, but simply a poorly nourished condition.
The column ‘ clinical character ’ gives the provisional diagnosis
arrived at by the physician in charge and by myself, either from
personal observation or from the information obtained from the
parents or friends of the patient. The data given in the preceding
columns are not always sufficient to bear out the diagnosis, which fact
is explained by the peculiar circumstances under which we worked,
especially that information often had to be obtained indirectly
and from a class of people who were in many cases unable to give
a definite answer to a concrete question, though a general impression
was conveyed by their statements. In view of the vagueness of our
opinion thus formed, the term diagnosis is avoided in the heading
of this column.
In the column ‘race,’ bl. means black, and col. coloured; no
cases were seen in whites, but the dictum that ‘ vomiting sickness ’
only occurs in the negro population was not confirmed, since two
cases were observed in coolie children; the one case in a chinaman
was of a different nature.
M. in the columns 18 and 19 means malarial parasites; P. means
Paraflasma jlavigenum.
The sign + in columns 19 and 20 means Diplococcus meningi¬
tidis; * indicates a growth of diplococci which differed from the
39 2
type of meningococci. The occurrence of other organisms, as
staphylococci and B. coli, in a few cases, was regarded as
accidental, and has not been tabulated.
In column 21, + indicates that albumin was found in the urine;
in nearly all these cases hyaline or granular casts were also
observed.
Most anatomical lesions are tabulated in accordance with the
observations made at the post-mortem examination, but have, as a
rule, been checked by histological observation. The statements
with regard to ‘ inflammation * in liver, pancreas, and kidney refer
exclusively to micro-cellular infiltration as observed by histological
observation, and, consequently, do not not always correspond to
the microscopical diagnosis of nephritis, etc., in the post-mortem
reports; it is well known that the term ‘acute parenchymatous
nephritis* does not necessarily imply that cellular infiltration has
already taken place.
Several organs, as thymus, thyroid body, pituitary body,
ovaries, and others, have been entirely omitted from the table, as
they never presented any lesions which appeared to have any
essential bearing upon the question of ‘ vomiting sickness *; they
were, however, automatically examined at the autopsies.
The post-mortem notes were dictated by me during the examina¬
tion and written down at once by the Medical Officer or by an
assistant.
The routine technique for histological examination was the
following: Small fragments of the organs were fixed, at the
autopsy, in sublimate alcohol (2 parts of saturated watery mercury
bichloride solution, and I part of absolute alcohol), and in
Flemming's strong osmic acid solution. Afterwards they were
embedded in paraffin. Sections of sublimate-alcohol specimens
were stained with iron-haematein, or with Friedlaender*s or Dela-
field's haematoxylin, and counterstained with Hansen’s aceto-picro-
fuchsin solution, or with eosin. Other sections from the sublimate-
fixed fragments were stained with Giemsa’s solution or with
Pappenheim’s ‘ Panchrom,* according to the technique previously
recommended (Seidelin, 1911, 1 and 2). In the case of the
Flemming technique sections were stained with safranin.
The organs usually preserved for histological observation were
393
liver, pancreas, kidney, spleen, lymph nodules, and myocardium.
Stomach, duodenum, adrenals, brain, and spinal cord were
frequently examined, and other organs occasionally, according to
special indications.
Examination for blood parasites was made in films of
peripheral blood and heart blood, and from organs as spleen, liver,
kidneys, and surface of brain. Microscopical examination was also
made of the centrifugalization-deposit of the spinal fluid.
Cultures were made from the fluid obtained by lumbar puncture,
in vivo or post mortem, or from the lateral ventricles of the brain,
at the autopsy, and in many cases from the heart blood. Ascites-
agar (nasgar) was the medium employed, and in addition to this,
in some cases haemoglobin agar or nutrient agar. Cultures were
considered negative if no colonies developed within a week.
A positive diagnosis of Diplococcus meningitidis was made, if
small, pale, greyish, semi-transparent colonies developed, consisting
of Gram-negative diplococci, which produced acid in glucose,
maltose, and galactose media, or changed at least one of these,
but produced no change in other sugars. Similar organisms, which
differed from the typical meningococcus in one or several particulars,
were frequently met with, and will be the object of special
discussion.
IV. DISCUSSION OF PERSONAL OBSERVATIONS
In order to discuss the cases tabulated in the preceding section,
it will be convenient to state the characteristics which would appear
to justify a diagnosis of ‘ vomiting sickness,’ with the exclusion of
other disease^. This means the provisional recognition of * vomiting
sickness ’ as a morbid entity, but the possibility must be left open
that this entity may fall to pieces afterwards if it is found on
further investigation that the cases may be classified under other
headings. As characteristic symptoms I would regard: sudden
onset; rapid development, the disease running its course to recovery
or death in less than twenty-four hours, as a rule, in children, and
in a few days in adults; vomiting; convulsions; coma; absence
of fever, or only low fever; collapse; in fatal cases, anatomical
lesions pointing to an acute intoxication, or hyperacute generalized
394
infection; negative result of the investigation for well-known
pathogenic organisms.
This provisional definition having been laid down, the cases fall
naturally into various groups. These groups will first be briefly
discussed, and particulars of typical cases afterwards be given as
illustrations of each group.
In the first group, A, all cases are included which correspond
more or less closely to the above definition. The cerebro-spinal
fluid has been examined in all cases, microscopically and by cultural
methods; the result was always negative, as far as the Diplococcus
meningitidis was concerned, but in some cases other microbes were
found, especially Diplococcus crassus and other meningococcus-
resembling diplococci. The blood was examined as far as possible,
and in the cases where such examination was made the result was
negative with regard to both malarial parasites and Paraplasm
flavigenum; in all other cases the diagnoses of malaria and yellow
fever were considered out of question, being unsupported or not
sufficiently supported by the clinical and (or) pathological evidence.
This group will, of course, form the principal object for the
following general discussion; in this place it shall only be pointed
out that the most constant and most remarkable pathological
changes were: fatty metamorphosis of liver, kidneys, and other
organs; necrobiotic changes of epithelia in pancreas, kidneys, and
liver; swelling and hyperaemia of lymph nodules; hyperaemia of
many organs, including the meninges, and a tendency to
haemorrhages; widespread oedema of the connective tissues. The
following cases belong to this group: Nos. 9, 14, 15, 16, 34, 37, 40,
41, 42, 46, 49, 52, 56, 59, 60, and 61; altogether 16 cases. Several
cases, provisionally classified under other headings, may also
belong to this group; case 58, for instance, closely resembles the
cases here included, and would have been classified accordingly but
for the reason that a few malarial parasites were found in the heart
blood, an occurrence which is very likely to be quite casual.
Notes from the cases in group A are given below, some in full
as types, others in abstract.
In the first case recovery took place; this case is, I believe,
typical of the mild form of ‘vomiting sickness.’
395
Case 9. A.S., 3 years, brown, 4, Chancery Lane, seen with Dr. Dryden in
the Public Hospital, Kingston. January 2i$t, 1913.
Admitted in a weak condition ; started vomiting at about 8 a.m., two hours
before admission. Vomited several times in hospital; the vomiting consisted,
as before, of abundant quantities of watery, frothy material, and took place without
any effort. The impression was most nearly that of an acute obstruction of the
pylorus. The child was stimulated, and reacted slowly in the course of the day,
without any specific treatment. He appeared well the following day, and was
discharged after two days, still somewhat weak and pale, but this seemed habitual.
Cerebro-spinal fluid was taken on the day of admission, at 2 p.m. Result:
Temperature normal. Pulse frequent, slightly irregular, soft. No abdominal
tenderness ; no diarrhoea. No meningeal symptoms.
There is a suspicion that the child had eaten ‘ physic nut ’ before taken ill,
but nothing definite is known, and the clinical picture did not correspond to that
described in cases of poisoning with Jatrapha curcas.
The following three cases, in three brothers, were seen by
Dr. Joslen, of Annotto Bay. The first patient was also examined
by me, and in the two other cases I performed the post-mortem
examinations with Dr. Joslen.
Case 14. A.N., 12 years, brown, born in Jamaica and has always lived there.
He was seen by Dr. Joslen at Evandale, St. Mary, where he lived in a poor hut,
with a leaking roof, in the bush, together with his father and two brothers. The
brothers were taken ill at the same time as A.N. (cases 15 and 16); the father was
unwell for about a day, complaining of fullness in the chest.
A.N. was taken ill, suddenly, on January 26th, 1913, at 10 p.m., with vomiting,
accompanied by considerable retching, slight epigastric pain, and slight headache.
He soon became weak and drowsy. Thus he was found on January 27th, at about
5 p.m., and next day he was taken to the Hospital in Annotto Bay, where he arrived
at 4.20 p.m. There I saw him the same evening, with Dr. Joslen.
January 27. Temperature 98*4° F. Pulse 120, regular, weak. Respiration
16, regular.
January 28, afternoon: temperature 98*4° F. ; pulse 120. Evening:
temperature 99 0 F.; pulse 88, regular, soft. No muscular rigidity; Kernig’s
symptom absent. Slight abdominal pain, corresponding to descending colon,
but no tenderness. No enlargement of liver or spleen. Heart dullness and heart
sounds normal. No sign of lung affection. The patient has no lice and no ticks.
January 29. Temperature normal; pulse 84, slightly irregular. The
patient has slept well, feels well, and is hungry.
The patient recovered.
Lumbar puncture was performed in the evening of January 28 ; about thirty
small drops of clear fluid were taken into two nasgar tubes, which remained sterile.
Blood smears were taken at the same time : they contained no parasites.
Case 15. C.N., 9 years, brown, born in Jamaica, and has lived there always.
Evandale, St. Mary. Brother to Cases 14 and 16. Good health until about
10 p.m., January 26th, 1913, when he suddenly started vomiting. He became
weak and drowsy, and died at about 5 —6 p.m., January 27th.
Symptoms as in Case 16.
Post-mortem examination at 8 p.m., January 28th, 1913. Body of poorly
nourished child. No jaundice. No rigor.
196
Lumbar puncture gives about thirty drops of clear fluid. Spinal membranes ,
both dura and pia, hyperaemic. Spinal cord shows no microscopical lesions.
Brain preserved unopened; the external aspect is normal. Dura mater
cerebralis normal, pia considerably hyperaemic.
Hypophysis cerebri of normal size and aspect.
Abdominal cavity contains no fluid. Peritoneum of normal aspect.
Abdominal organs normally situated.
Spleen: 18 x n X J cms. Capsule smooth, transparent. Colour reddish
grey. Consistence diminished, slightly friable, not soft.
Right kidney : 9x^x2 cms. Capsule easily detached. Surface smooth,
without hyperaemia and haemorrhages. Consistence normal. Cortical substance
pale. Slight hyperaemia of pyramidal bases.
Left kidney as right.
Mucosa of renal pelves, ureters, and bladder normal.
Genital organs normal.
Liver: 30 x 14 X 7 cms. Left lobe very long, which accounts for the
excessive length of the organ, which is not essentially enlarged. Capsule smooth,
transparent. Colour greyish red, with some paler, but not distinctly yellowish
patches. The tissue is more uniformly grey. Consistence normal.
Pancreas : 12 cms. long, slightly hyperaemic ; consistence soft.
Suprarenal capsules of normal aspect.
Pleural cavities contain no fluid. There are extensive, but delicate, adhesions
of right pleura and similar, but less developed, of the left. Lungs hyperaemic in
postero-inferior portions, contain air throughout. No infiltrations, no oedema,
no infarcts. Slight anthracosis.
Bronchial lymph nodules slightly enlarged, anthracotic, and some of them
fibrous, but no definite tuberculous phenomena are met with in lymph nodules
or in lungs.
Larynx , trachea , and bronchi normal, except that frothy fluid escapes from
the bronchi.
Tongue slightly coated.
Tonsils enlarged and the right considerably hyperaemic.
Oesophagus normal.
Thyroid body normal.
Pericardium contains no fluid. No pericardial, nor endocardial haemorrhages.
Heart-conus: 6 x 8 x 2J cms. Left ventricle fairly well contracted. Endocard
and all valves normal. Myocard pale, of slightly diminishing consistence, shows
fine yellowish stripes.
The stomach contains a considerable amount of brownish fluid, apparently
not haemorrhagic, but corresponding to intestinal contents. The mucosa shows
considerable hyperaemia, and on the anterior surface, near to the lesser curvature,
are observed several petechiae.
The duodenum shows hyperaemia, but no haemorrhages. At a distance of
about 2 cms. below the pylorus an ulcer is observed, which is of oval shape, 8 mms.
long and 6 mms. wide, and is situated on the posterior wall. The ulceration has
penetrated the mucous and muscular coats, and the bottom is formed only by the
retro-duodenal tissues. The borders of the ulcer are slightly infiltrated, but there
is no overlapping of the edges. The jejunum , ileum 9 colon and appendix without
pathological phenomena.
Bile ducts and gaU-bladder free from stones and obstructions; the gall-bladder
contains greenish bile.
397
Numerous ascarides are present, about 35 in the small intestine, and one in the
stomach. In the coecum a few Tricocephali are present.
Anatomical diagnoses: Hyperaemia meningum. Hyperplasia lienis. Hypo¬
stasis pulmonum. Metamorphosis adiposa hepatis. Pancreatitis parenchymatosa
acuta. Sequelae pleuritidis duplicis. Gastritis acuta cum petechiis. Ulcus
duodeni. Helminthiasis intestinalis.
The histological examination of the organs was of no particular interest, as
post-mortem changes were marked. It may be mentioned that sections of the
liver showed intense fatty change, which was irregularly distributed throughout
the lobules ; the tissues were anaemic, where fatty change was most marked, in
other places capillary hyperaemia was observed. The pancreas showed marked
oedema of the stroma and partial necrobiosis of epithelia. The myocard showed
fragmentation and oedema of stroma.
A short. Gram-negative bacillus developed in the nasgar-tubes, inoculated
with spinal fluid; no cocci.
Case 16. C.N., si years, brown, born and lived in Jamaica, Evandale,
St. Mary. Brother to A.N. and C.N., Cases 14 and 15.
THe patient was seen by Dr. Joslen, Annotto Bay, January 27th, 1913, and
the post-mortem examination performed by me on January 28th.
Apparently good health until January 26th, 10 p.m., when he suddenly started
vomiting, and went on vomiting until death. Drowsy, listless and* weak. Died
at 5—6 p.m., January 27th.
No desire for food, but thirst. No chilliness, rigor, convulsions. No eye
symptoms. Skin natural, no eruption, no jaundice. No fever. No lung
symptoms. No headache. Perhaps slight cardialgia; considerable retching.
No delirium, irritability, paralysis, rigidity, nor retraction of neck.
Post-mortem examination at 8 p.m., January 28th, 1913.
Body poorly nourished. Rigor mortis is present. No lice are observed.
There is no jaundice. Superficial ulcerations are observed on both lower extremi¬
ties.
Lumbar puncture gives 15 drops of blood-stained fluid.
The spinal meninges , both pia and dura, are hyperaemic, and the spinal cord
is also somewhat hyperaemic.
The cerebral dura is normal, but the pia is considerably hyperaemic, especially
on the convex surface of the brain. There is no fibrinous exudation. The lateral
ventricles are not distended. There is no hyperaemia, and no other pathological
lesions of the brain substance.
The hypophysis cerebri and the thyroid body are normal.
The larynx , trachea , and bronchi are normal. No pleural fluid, nor adhesions.
The lungs show hyperaemia in their postero-inferior portions, but neither
oedema, nor infiltrations, nor infarcts; there is slight anthracosis. The bronchial
lymph nodules are slightly enlarged and anthracotic.
The heart conus is 5 x 6 x 3 cms. The left ventricle is well contracted.
The pulmonary artery contains no thrombi. All the valves are normal, as is the
whole of the endocardium, and the pericardium. The myocardium is pale, other¬
wise normal. The pericardium contains a few cubic centimetres of clear fluid.
The inguino-crural lymph nodules are slightly enlarged, the axillary not enlarged.
The abdominal cavity contains no fluid ; the peritoneum is of normal aspect.
The abdominal organs are normally situated, with the exception of the kidneys.
The sigmoid flexure is considerably distended with faeces.
39 8
The mesenteric lymph nodules are slightly enlarged ; they show no haemorrhages,
and no signs of tuberculosis.
The diaphragm reaches on the right side the 5th, on the left side the 4th rib.
The spleen is 11 X 7 X 2 cms. Capsule smooth, transparent; colour dark
greyish red; no haemorrhages; consistence soft; follicles grey, somewhat
prominent. A supernumerary spleen is found in the gastro-splenic ligament.
The suprarenal capsules are normal.
The kidneys are united at their inferior extremities in the shape of a horse-shoe,
with two separate pelves on the anterior surface. The capsule is easily detached.
The surface is smooth, without any vascular distension. The renal tissue is normal;
there is slight hyperaemia of pyramidal bases. The mucosae of the renal pelves,
ureters and bladder are normal. The bladder contains a considerable quantity
of pale urine. The genital organs are normal.
The liver is 18 x 12 x 5 cms. The capsule is smooth and transparent;
colour uniformly red ; substance reddish grey without haemorrhages, and with
no yellow patches. Consistence normal.
The pancreas is 10 cms. long, hyperaemic, and of diminished consistence.
The stomach contains alimentary substance and some mucus, but no blood.
There are no haemorrhages in the mucosa.
Oesophagus , normal. Tonsils without signs of acute inflammation. Tongue
slightly coated.
The intestines present no pathological changes. A large number of ascarides
are present, but no other parasites.
The bile ducts are patent; the gall bladder contains a considerable amount
of greenish bile ; mucosa normal.
Anatomical diagnoses: Hyperaemia meningum. Hypostasis pulmonum.
Lymphadenitis bronchialis, mesenterialis et inguinalis. Splenitis acuta. Pan-
creatitis parenchymatosa acuta. Lien accessorius. Symphysis renum. Helmin¬
thiasis intestinalis.
As in the foregoing case, post-mortem changes interfered with the histological
examination. Sections of an inguinal lymph nodule showed marked hyperaemia
and some lymphocyte-infiltration of the capsule. Sections of the spinal cord
showed hyperaemia, but no cellular infiltration of the pia, and, apparently, oedema
of pia and cord.
Short, Gram-negative bacilli develop in the nasgar tubes, inoculated with
the spinal fluid; no cocci.
The following is an example of vomiting sickness in a child of
East Indian race, though, I believe, born in Jamaica. In this case,
the presence of jaundice aroused suspicion of yellow fever, but after
examination of stomach and liver this diagnosis was entirely
dismissed.
Case 34. R.S., 4 years, coolie child, Windsor Park, St. Catherine, seen, after
death, with Dr. A. G. Curphey, at the Hospital, Spanish Town, on February 6,1913.
The child was admitted this morning, at 10 a.m., in a dying condition. He
was said to have been taken ill suddenly the previous day, February 5, at 3 p.m.,
with vomiting. The vomits consisted of frothy, watery fluid, and were repeated
the following morning. The child was crying and restless, but no shrieks were
noticed. He had been healthy until this illness. In the hospital clonic convulsions
were observed with intervals of about five minutes. There was no strabismus;
399
no rigidity of neck ; no Kernig’s symptom. Temperature 101*4°. The pulse was
rapid, about 140, weak, regular. Frothy fluid escaped from the mouth, no
vomiting took place in the hospital. A yellow stool was obtained after enema.
The child died at 11.15 a.m. (Notes by Dr. Curphey, as I arrived not until about
2.30 p.m.).
The mother reports that another child of hers died suddenly on February 1,
after having vomited watery, frothy fluid.
One child alive and in good health.
Post-mortem examination , at 3 p.m., on February 6, about four hours after
death. Well nourished body. Rigor present. Slight jaundice. Slight enlarge¬
ment of lymphatic nodules in neck, axillae, and inguina.
Lumbar puncture gives clear fluid, of which eight drops are taken in nasgar tube,
and two smears are made. Spinal cord not examined. Dura mater cerebralis
normal, pia slightly hyperaemic in posterior part of convex surface. No distension
of lateral ventricles. Brain substance macroscopically normal throughout.
Hypophysis cerebri and thyroid body of normal size and aspect.
The pleural cavities contain no fluid, and there are no pleural adhesions.
The lungs show slight hyperaemia of postero-inferior portions, but are otherwise
normal. The bronchial lymph nodules are slightly enlarged and anthracotic.
The bronchi, trachea, and larynx are normal.
The pericardium contains no fluid, and shows no petechiae. The heart is of
normal size. The pulmonary artery is free. The endocard and all the valves are
normal, the myocard is pale, greyish, of normal consistence.
The peritoneum is of normal aspect, and contains no fluid. Abdominal organs
normally situated.
The tongue is slightly whitish coated. The tonsils are not enlarged. The
oesophagus is normal. The stomach shows normal conditions ; there is no hyper¬
aemia of the mucosa, and no haemorrhages. The duodenum shows considerable, and
the jejunum slight swelling of lymphatic follicles ; no other abnormal conditions
of intestinal tract. The bile ducts are patent; the gall bladder contains a small
quantity of greenish bile.
Liver: 20 x 11 X 5 cms. Capsule smooth, transparent; surface reddish,
with grey and yellowish patches; the consistence is normal in the red portions,
but diminished in the yellowish ones. The structure of the organ is homogeneous,
without distension of vessels.
The pancreas shows slight hyperaemia, but is of normal size and consistence.
The spleen measures 6 x 4 X ii cms. Its capsule is smooth and transparent,
its colour dark greyish red, and its consistence normal. The follicles are grey and
prominent.
There is a slight hyperaemia of the left suprarenal capsule , but not of the right
one, and both organs are otherwise normal.
The left kidney is 7 x 4 X 2 cms. The capsule is easily detached, the surface
is smooth, dark, and shows capillary, but not venous, hyperaemia. The renal
substance is dark red, without haemorrhages, and without yellow patches. The
right kidney shows conditions similar to those of the left, except that the colour
is slightly more pale.
The mucosae of renal pelves, ureters, and bladder are normal. The bladder
contains about 50 c.c. of clear, yellow urine.
The genital organs are of normal aspect.
The urine is pale, clear, acid, contains a considerable amount of albumen, and
a few casts, besides epithelial cells; it contains no sugar, no bile pigment, no
erythrocytes, and no leucocytes.
Anatomical diagnoses: Hyperaemia piac matris cerebri l.g. Hypostasis
pulmonum l.g. Duodenitis follicularis. Jejunitis follicularis l.g. Lympho-
adenitis mesenterialis, bronchialis, cervicalis, axillaris et inguinalis simplex. Meta¬
morphosis adiposa hepatis. Hyperaemia pancreatis, glandulae suprarenalis sinistrae,
et renum. Icterus.
Histological examinations :—
Liver: Marked capillary hyperaemia, slight microcellular infiltration of
periportal connective tissue. No necrobiosis, and only very slight vacuolization
of cells.
Pancreas : Marked venous and capillary hyperaemia, slight oedema of stroma,
small patches of necrobiotic cells.
Spleen: Diffuse hyperaemia.
Kidney: Hyperaemia, which is very irregularly distributed, and often very
intense ; in a few places some interstitial leucocyte-infiltration ; slight vacuolization
of basal portions of cells in some of the convoluted tubules ; in the same kind of
tubules necrobiotic changes are common, with granular appearance of the proto¬
plasm and more or less advanced karyolysis.
Duodenum : Enormous hyperplasia of Brunner’s glands which not only increase
the thickness of the submucosa, but also occupy large parts of the mucosa, even
dislocating and compressing the Lieberkuhn’s crypts, and in some places reaching
the free surface. There is also considerable hyperplasia of the lymphoid follicles
and diffuse lymphoid infiltration of the mucosa.
Axillary lymph nodule: Diffuse hyperaemia and slight oedema of stroma.
Brain : Slight hyperaemia, no microcellular infiltration.
Spinal fluid ; Very few cells, chiefly erythrocytes; some mononuclears, and
a few polymorphonuclears.
Microbiological examination :—
Heart-blood: No growth.
Spinal fluid: Growth of a diplococcus, which forms large groups. It
decolourizes with some difficulty by Gram’s and Claudius’s methods. It produces
acid in glucose, maltose, galactose, lactose, and saccharose media.
Spleen: Growth of diplococcus identical with the one in the spinal fluid.
No parasites are found in smears from heart blood, liver, spleen, and spinal
fluid.
In the following case there was no vomiting, but otherwise it
appears to belong to the type of 1 vomiting sickness.’ There were
no signs of meningitis or yellow fever, clinically or anatomically.
No growth was obtained from the spinal fluid, and none in nasgar
tubes from heart blood, but on haemoglobin-agar inoculated with
heart blood there was a growth of diplococci, otherwise similar to
meningococci, but showing the striking difference, that the colonies,
both on haemoglobin-agar, and in subcultures or nasgar, were of a
saturated whitish appearance.
Case 40. A.M.L.Y., z\ years, black. Barbican, near Cherry Garden Gate,
seen on February 12, 1913, with Dr. Croswell.
The child has enjoyed good health until this morning. She awoke at 6 a.m.,
apparently well, asking for water. Soon after, however, she complained of pain
401
in her stomach, and was seized by convulsions, apparently a kind of slow clonic
contractions. She has suffered from no previous disease ; in particular, she has
had no cough, no fever, and no symptoms of worms. She was seen by Dr. Croswell
at 8 a.m.; she was then comatose, and there was complete muscular relaxation.
There was no abdominal retraction, no rigidity, and no Kernig’s symptom.
Temperature 97*8° F. No jaundice. The action of the heart was fairly strong.
Injections of strychnin, digitalis, and nitroglycerin were given.
The child was in a dying condition, when I arrived at 10.45 a.m. The
respiration was superficial, the pulse weak, but regular; it became immediately
filiform, and could not be counted, but was not very frequent. There were no
convulsions, and no rigidity, especially not of the neck, and no Kernig’s symptom.
The arms were faintly contracted in a semi-flexed position, but immediately
afterwards became relaxed. The teeth were closed. There was grinding of the
teeth. The child gave a faint shriek, and died at 10.50 a.m. At the moment of
death, urine was passed in considerable quantity. The abdomen was considerably
distended and this distension was said to have developed during the last ten minutes.
There was no enlargement of liver or spleen.
About an hour after death spinal fluid and heart blood were taken in nasgar
and haemoglobin-agar tubes, and smears were made.
Post-mortem examination (with Dr. Edwards, at II a.m., on February 13, about
twenty-four hours after death). Well nourished, healthy looking body.
Dura mater cerebralis , and, particularly, spinalis considerably hyperaemic ;
pia mater slightly hyperaemic, mostly pronounced on convex surface of the brain
and on the lumbar portion of the cord. No inflammatory exudation, and no free
fluid in the lateral ventricles. The brain and spinal cord are of normal appearance.
The inguinal, axillary, and cervical lymph nodules are slightly enlarged, and
intensely hyperaemic.
The abdominal cavity contains no fluid. The peritoneum is of normal aspect.
The abdominal organs are normally situated. The intestines are considerably
distended. The diaphragm reaches the fourth intercostal space on both sides.
The spleen measures 7x5x2 cms. The capsule is smooth and transparent,
the colour dark red and grey. The follicles are grey and very prominent, the
consistence normal.
Suprarenal capsules normal.
Right kidney : 6 x 4 X if cms. Capsule easily detached ; surface smooth,
with venous and capillary hyperaemia ; cortex hyperaemic, with yellowish grey
patches, in its central portions; pyramids pale. Left kidney shows the same
conditions as the right.
Renal pelves, ureters, and bladder of normal appearance. The bladder
contains a small quantity of pale urine.
Genital organs normal.
Mesenteric lymph nodules considerably enlarged and slightly hyperaemic.
Liver: 23 X 13 X 4 cms. Capsule smooth, transparent ; colour dark purple ;
substance uniformly reddish grey ; consistence normal.
The pancreas is hyperaemic and of soft consistence.
The stomach contains a considerable quantity of bile-stained mucus. The
mucosa is hyperaemic, but without haemorrhages.
The duodenum is hyperaemic. The jejunum shows a few hyperaemic patches
and slight swelling of both solitary follicles and Peyer’s patches. The swelling of
the patches increases considerably in the ileum, especially in its terminal portion
and is here accompanied by hyperaemia. The colon shows throughout its whole
402
length follicular swelling and on the top of each swollen follicle a black, minute spot.
There are no parasites in the intestinal tract, except one small ascaris in the ileum;
in particular no parasites arc seen corresponding to the swollen follicles in the
colon. The bile ducts, gall bladder, and pancreatic ducts are of normal aspect.
The gall bladder contains a small quantity of greenish bile.
The oesophagus is normal.
The tonsils are considerably swollen and hyperaemic, with haemorrhages in
the tissue.
The tongue is covered, on the whole surface, with whitish fur.
The larynx is slightly hyperaemic, the trachea and the larger bronchi more
pronouncedly so, and in the smaller bronchi, the mucous membrane is both swollen
and hyperaemic, whilst there is an abundant mucous secretion. There is a pro¬
nounced hyperaemia of the postero-interior portions of both lungs , whilst the
anterior portions are markedly pale.
The bronchial lymph nodules are slightly enlarged and hyperaemic, as well as
the mediastinal. There are no signs of tuberculosis in either lungs or lymph
nodules.
The pleurae are normal.
The pericard y endocard , and valves are normal. The myocardium is very pale.
The hypophysis cerebri and the thyroid body are normal.
The thymus measures 5 x 4 X \ cm.; it is of normal appearance and
consistence.
Anatomical diagnoses: Lymphadenitis acuta. Tonsillitis haemorrhagica.
Enteritis follicularis. Colitis follicularis et pigmentosa. Lymphadenitis mesen-
terialis m.g. Hyperaemia ventriculi, renum et meningum. Pancreatitis acuta
parenchymatosa. Splenitis acuta follicularis. Hypostasis pulmonum. Bronchitis
capillaris. Bronchitis, tracheitis et laryngitis incipiens. Anaemia myocardii.
Helminthiasis intestinalis ( one ascaris).
Urine : Contains small quantity of albumen and some hyaline casts.
Histological examination :—
Axillary and cervical lymphatic nodules: intense hyperaemia, venous and
capillary. Moderate oedema of stroma. Mesenteric lymphatic nodules: oedema
of stroma, but no hyperaemia.
Spleen: Intense diffuse hyperaemia.
Liver : Intense diffuse hyperaemia, moderate fatty changes equally distributed
throughout the lobules, slight necrobiotic changes.
Microbiological examination :—
Nasgar tubes inoculated with spinal fluid and heart blood remain sterile, except
one in which one colony of short Gram-negative bacilli develops.
Haemoglobin agar tubes inoculated with spinal fluid remain sterile, but not
those inoculated with heart blood , in which saturated white colonies develop,
consisting of Gram-negative cocci, mostly diplococci which grow abundantly,
also in nasgar, and which ferment glucose, maltose, and galactose, but not lactose
and saccharose.
The heart blood contains microscopically no organisms ; smears from liver,
kidney, spleen and mesenteric lymph nodules contain various types of bacteria and
those from liver and kidney contain also small diplococci. Smears from bone
marrow contain no organisms.
Case 41. M.A.R., 3 years, black, Whitehall, with Dr. C. R. Edwards,
Half-way Tree.
4 ° 3
Seen alive by Dr. Edwards, not by me. Previous health good. Taken ill
in the morning of February 13, 1913, with vomiting and convulsions; died at
11 a.m. on the same day.
Post-mortem examination , at 11 a.m., on February 14. Body fairly well
nourished. No cutaneous affections, nor parasites. Rigor present. No jaundice.
On the skin of the back a number of irregular, black patches, as from burns, and
on the left hip a bulla, 5x2 cms.
Cervical , axillary , and inguinal lymph-nodules swollen and intensely
hyperaemic.
Spinal fluid taken after death by Dr. Edwards, clear ; only a small quantity
was obtained. On opening the spinal canal the fluid was found somewhat thickened,
slightly gelatinous. Cerebral dura shows slight, and spinal dura moderate
hyperaemia. Spinal and cerebral pia shows considerable hyperaemia, mostly
pronounced on the convex surface of the brain. No distension of the lateral
ventricles. Cerebral and medullar substance normal.
Hypophysis cerebri normal.
Diaphragm on the right side at fourth, and on the left at fifth intercostal
space.
Pleurae without fluid and adhesions. The Lungs are slightly hyperaemic,
chiefly in postero-inferior portions. They contain several small, haemorrhagic
infarcts, and numerous petechiae are seen on the pleura of the right inferior lobe.
There is slight oedema of the inferior lobes of both lungs.
The bronchial lymph nodules are slightly enlarged and slightly hyperaemic.
The smaller bronchi show hyperaemia and swelling of their mucosa and
considerable mucous secretion ; similar conditions obtain in the larger bronchi,
trachea, and larynx.
Thyroid body normal.
Pericardium , pulmonary artery , and aorta normal. Epicardium hyperaemic.
Left ventricle well contracted, forms apex. Heart conus 5x6x2 cms.
Endocard and valves normal. Myocard pale, of slightly diminished consistence.
Peritoneum normal, contains no fluid, abdominal organs normally situated.
Appendix occupies a posterior, lateral, ascending position.
Mesenteric lymph nodules considerably enlarged and slightly hyperaemic.
Spleen: 7x5x2 cms. Capsule normal; colour dark purple, with grey
spots ; follicles slightly prominent, grey ; consistence normal.
Suprarenal capsules normal.
Right kidney: 8x4x2 cms. Capsule easily deatched. Surface dark,
with capillary hyperaemia ; cortex hyperaemic, with yellowish grey patches in
central portions. Pyramids pale. Left kidney shows the same characters.
Mucosa of renal pelves and ureters hyperaemic. The bladder contains a
considerable quantity of pale yellow urine, of which some part is collected ; the
mucosa is normal.
The oesophagus is hyperaemic. The stomach contains a small quantity of a
alimentary fluid; the mucosa shows slight hyperaemia. The duodenum shows
considerable hyperaemia and some petechiae. The jejunum shows numerous
hyperaemic patches and a few ecchymoses in its upper portions. Ileum pale, with
considerable swelling, but no hyperaemia of Peyer’s patches. Appendix 10 cms.
long, normal. The large bowel shows a few hyperaemic patches, is otherwise
normal. The jejunum, ileum, and colon contain an enormous number of guava
seeds; also a few in appendix. One Tricocephalus in coecum. No other parasites.
Bile ducts and gaU bladder are patent and of normal aspect.
4°4
The liver measures 22 x 11 X 4 cms. Capsule normal ; colour purple grey,
with a few yellowish patches ; consistence and structure normal.
The pancreas is hyperaemic and of dimished consistence.
The tongue is covered on its anterior portion with a whitish fur, on the posterior
part of its surface with a dark and slightly haemorrhagic coating.
The tonsils are enlarged, hyperaemic, and show in their tissue haemorrhages
and purulent infiltration.
The thymus measures 5 X 3 X J cms., and shows normal aspect.
The urine is pale, clear, contains a small quantity of albumin and a few granular
casts ; no sugar, no bile pigments.
Anatomical diagnoses: Poly-lymphadenitis acuta. Sequelae combustionis.
Hyperaemia men ingum. Hyperaemia et infarct i haemorrhagici pulmonum.
Petechiae pleurae dextrae. Laryngitis, tracheitis, bronchitis et bronchitis capil-
laris catarrhalis. Nephritis parenchymatosa acuta. Hyperaemia ventriculi et
intestinorum. Ecchymoses duodeni et jejuni.
Histological examination :—
Liver: Intense hyperaemia of a diffuse character; marked fatty change;
slight necrobiosis and slight microcellular infiltration of stroma.
Pancreas : Almost complete necrobiosis of cells, the nuclei not staining with
haematein, and the cell limits badly defined.
Kidney : Intense necrobiosis of epithelia of convoluted tubules, the protoplasm
of which is granular and the nuclei poorly stained or entirely unstained.
Glomeruli and straight tubules fairly well preserved.
Myocardium : No changes observed in sections.
Lymph nodule: Diffuse hyperaemia ; patches of necrobiosis.
Spinal cord : Pia mater shows oedema and patches of microcellular infiltration.
Tonsil: Localized necrosis and haemorrhages.
Microbiological examination :—
Large Gram-positive bacteria develop in cultures on nasgar from the spinal
fluid; similar bacteria are seen in smears from the spleen, and in sections from
various organs. No diplococci, in smears, or cultures. Culture-tubes (nasgar
and haemoglobin-agar) inoculated with heart blood remain sterile.
In the following case, the clinical symptoms suggested a diagnosis
of meningitis ; this diagnosis, however, was not confirmed
anatomically or microbiologically ; the case may provisionally be
referred to as one of ‘ meningismus.’
Case 46. E.B., 2 \ years, black, Rodney Street, Admiral Town (near Kingston),
seen at Public Hospital, Kingston, with Dr. Thomson.
Sudden onset of convulsions at about 7 a.m., February 17,1913. No vomiting.
Admitted to hospital at 9.45 a.m.; temperature 98° F.; patient unconscious;
pupils dilated. Limbs rigid; head retracted, marked opisthotonus. Kemig’s
sign present, most marked on the right side.
Lumbar puncture made (Dr. T.) : Water-clear fluid escapes under considerable
pressure.
When examined later on, by Dr. T. and myself, at 12.30 p.m. the limbs are
absolutely limp, and there is only a faint indication of Kernig’s sign on the right
side,.whilst it is entirely absent on the left. Still later, it is absent on both sides.
Pulse fairly strong, but very irregular, about 100.
Several convulsions occurred in the hospital, consisting chiefly in tonic spasms,
and the child died at 4.30 p.m.
Post-mortem examination , at 11.30 a.m., on February 18, nineteen hours after
death. Body well nourished. No skin affections, nor parasites. No rigor. No
jaundice.
Cervical , axillary , and inguinal lymph nodules slightly enlarged and hyperaemic.
The subdural space contains a considerable amount of serous fluid at the basis
of the cranium, and fluid is seen escaping from the spinal canal. Slight hyperaemia
of cerebral dura ; spinal dura normal. Pia slightly hyperaemic, mostly so on the
convex surface of the brain, and on the lumbar portion of the cord. No fibrinous
exudation. The lateral ventricles are not dilated, and contain only a few drops
of clear fluid. Brain and cord substance normal.
Hypophysis cerebri somewhat hyperaemic, not enlarged.
Pleurae normal. Both lungs contain a few small haemorrhagic infarcts, are
otherwise perfectly normal, without hypostasis, and without anthracosis. Bronchi,
small as well as larger, trachea, and larynx, normal. Bronchial lymph nodules
moderately enlarged and hyperaemic, not anthracotic.
Thymu j : 5 x 2 x 0*3 cms., of normal appearance.
Thyroid body normal.
Pulmonary artery and aorta normal. Pericardium contains a few c.c. of clear
fluid. Left ventricle well contracted, forms apex. Heart conus 4i X 5J X 2 cms.
Epicard, endocard, and valves normal. Myocard slightly pale, and of slightly
diminished consistence.
Diaphragm at fourth intercostal space on right, and at fifth on left side.
Peritoneum normal, without fluid. Abdominal organs normally situated.
Spleen : 10 x 5 X 2 cms. Capsule normal; colour pale, with some dark
patches, and a number of petechiae on the surface. Follicles prominent, light
grey. Consistence normal.
Suprarenal capsules slightly hyperaemic, otherwise normal.
Right kidney : 6 X 4 X I i cms. Capsule easily detached ; surface smooth,
dark, shows capillary hyperaemia ; cortex grey ; pyramids slightly hyperaemic at
bases, otherwise pale; consistence normal. Left kidney: 6x4x2} cms.,
otherwise as the right. Renal pelves, ureters, and bladder normal; bladder
contains a small quantity of clear urine.
Genital organs normal.
Liver : 20 X 12 X 5 cms. Capsule normal; colour of surface dark purple,
with some yellow patches ; substance grey, likewise with yellow patches ; con¬
sistence normal.
Pancreas slightly enlarged, intensely hyperaemic, and of somewhat diminished
consistence.
Tongue normal. Tonsils slightly enlarged, not hyperaemic. Oesophagus
normal.
Stomach contains alimentary fluid; mucosa normal.
Duodenum shows slight hyperaemia and swelling of follicles.
Jejunum normal. The lower part of the ileum shows considerable swelling
of both solitary follicles and Peyer’s patches ; the latter are hyperaemic. Appendix
7 cms., normal. Colon normal. In the lower part of the ileum two ascarides ;
no other parasites.
Gall bladder and bile ducts normal; the former organ contains greenish bile ;
the latter are patent.
Mesenteric lymph nodules considerably enlarged, not hyperaemic.
Anatomical diagnoses: Poly-lymphadenitis, l.g. Hyperaemia meningum.
Infarcti haemorrhagici pulmonum. Hyperaemia renum. Pancreatitis parenchy-
matosa acuta. Metamorphosis adiposa hepatis. Petechiae lienis.
406
Histological examination :—
Lymph nodules : slight hyperaemia. Moderate oedema of stroma.
Spleen : Marked oedema of stroma.
Liver : Intense fatty changes, moderate necrobiosis, both phenomena equally
distributed throughout the lobules. The cells, when their limits are recognizable,
are seen to contain a considerable number of large vacuoles (fat), but are in no
case filled to distension with a single fat drop, nor do they contain large numbers
of minute droplets. No hyperaemia. No inflammatory phenomena. Moderate
oedema of stroma.
Kidney: Patches of intense hyperaemia ; epithelia of convoluted tubules in
many places granular, and nuclei poorly staining or even absent, though the tubules
are fairly well preserved. Glomeruli of normal appearances. Slight oedema of
stroma. In osmic acid-preparations small fat droplets are seen in the epithelia of
the convoluted tubules.
Brain: No hyperaemia.
In films made from the spinal fluid without centrifugalization no cells are seen.
After centrifugalization very few cells which are mostly lymphocytes, also several
polymorphonuclear leucocytes.
Microbiological examination :—
Nasgar tubes inoculated with spinal fluid; one tube remains sterile and in
another tube there is growth of ( a ) Gram-positive staphylococci, (i b ) Gram-negative
diplococci forming chains, and changing maltose, but not glucose, galactose,
saccharose and mannite. Heart blood : Nasgar tubes remain sterile ; on haemo¬
globin agar Gram-negative cocci develop similar to those in cultures of spinal fluid
and showing marked formation of chains, but less marked diplococcal arrangement.
No parasites are found on microscopical examination of spinal fluid, peripheral
blood, heart blood, spleen and bone-marrow.
Case 49. M.A.W., 3 years 2 months, black, seen after death, with Dr. Earle,
May Pen, who had seen the child on the previous day, when it was ill.
February 19, 1913.
The child was quite well on February 17, and went to bed in the evening as
usual, and slept well. On the 18th about 7 a.m. she was taken ill with convulsions
which started suddenly; there were contractions of arms and legs and rolling of
the eyes; no stiffness of neck observed. The attacks of convulsions followed each
other rapidly. She was unconscious. She was seen by Dr. Earle at about 10 ajn.
She was afterwards conscious and quiet until about 3 p.m., when convulsions again
set in—about five occurred during the afternoon—and she again became unconscious
and remained so until death, which took place at 9.45 p.m. She vomited about
five times during the day, each vomiting occurring when a convulsion was about
to set in.
Post-mortem examination , at I p.m., about fifteen hours after death. Body
of well-nourished, absolutely healthy-looking child. No rigor. No jaundice.
No cutaneous affection, and no parasites.
Spinal fluid flows freely by lumbar puncture, about 5 c.c. taken for examination ;
it is perfectly clear.
The spinal venous plexus are extremely gorged; dura and pia spinalis are
moderately hyperaemic, pia especially in its lumbar portion ; no fibrinous exudation.
Cord substance normal.
Dura mater cerebralis normal; pia moderately hyperaemic, especially on
posterior portion of the convex surface. No exudation. Lateral ventricles
contain a few c.c. of clear fluid. Brain substance normal.
Cervical , axillary and inguinal lymph nodules moderately enlarged and slightly
hyperaemic.
Diaphragm at fourth intercostal space on both sides.
Pleurae normal. Lungs contain a few small, haemorrhagic infarcts, and are
slightly hyperaemic in postero-inferior portions; otherwise normal. The smaller
as well as the larger bronchi, trachea, and larynx, contain a small amount of frothy
mucus; otherwise perfectly normal.
The pericardium contains a small amount of clear fluid; heart conus
5x6x3 cms. Pulmonary artery and aorta normal. Epicard, endocard, and
valves normal. Myocard slightly pale, of somewhat pasty consistence.
Thymus : 9X 5 X i cms. Appearance normal.
Thyroid body normal.
Peritoneum normal, without fluid. Position of abdominal organs normal;
appendix inwards and downwards into the pelvis. Mesenteric lymph nodules
moderately enlarged, not hyperaemic.
Spleen : ioxixiJ cms. Capsule normal; colour pale, with dark spots;
follicles grey, slightly prominent; consistence normal.
Suprarenal capsules hyperaemic, of normal size.
Left kidney : 9x4x2 cms. Capsule easily detached ; surface pale, with
somewhat dilated stellate veins; cortex grey with yellowish patches ; pyramids
slightly hyperaemic at their basis, otherwise normal. Right kidney, 8 X4 X1J cms.,
similar to the right one, but slightly more hyperaemic, and with more prominent
yellowish patches.
Renal pelves hyperaemic, ureters and bladder normal; the bladder contains
about 30 c.c. of dear urine.
Genital organs: hyperaemic, otherwise normal.
Liver : 20 x 12 x 5 cms. Capsule normal; colour of surface distinctly
yellowish with a few hyperaemic patches ; tissue uniformly yellowish grey;
structure normal; blood filling about normal; consistence somewhat diminished,
pasty.
Pancreas: n cms. long; not hyperaemic; consistence considerably
diminished.
The stomach contains a small quantity of alimentary mucus; the mucosa
shows a few small patches of capillary hyperaemia.
Duodenum normal, as well as jejunum and ileum.
Colon shows a large number of black dots, corresponding to the centres of the
follicles, which are not swollen.
The colon ascendens contains one Tricocephalus and two ankylostomes; no
ankylostomes higher up in the intestine.
Bile ducts and gall bladder normal.
The whole surface of the tongue is covered with whitish fur.
Tonsils normal.
Oesophagus normal.
Anatomical diagnosis : Hyperaemia meningum. Poly-lymphadenitis. Hypo¬
stasis et infarcti haemorrhagici pulmonum. Metamorphosis adiposa myocardii et
hepatis. Hyperaemia capsularum suprarenalium. Nephritis parenchyma tosa
acuta duplex l.g. Pancreatitis parenchymatosa acuta. Hyperaemia ventriculi l.g.
Pigmentatio coli. Helminthiasis intestinalis.
Histological examination :—
Liver: Intense fatty metamorphosis, in some places most intense around
portal vessels, in others equally distributed over all zones of the lobules. Capillary
hyperaemia irregularly distributed. Slight necrobiotic changes.
408
Pancreas : No hyperaemia or cellular infiltration. No fatty change. The
epithelia are as a rule normal, but in isolated places very poorly stained, the nuclei
being almost invisible. Langerhans’s islets well preserved and very definite.
Kidney : Some of the convoluted tubules show advanced granular and fatty
metamorphosis, but in most the cells are fairly well preserved and the nuclei well
stained. The glomeruli are considerably retracted.
Myocardium : slight fragmentation.
Spinal fluid : Numerous cells, mostly mononuclear ; only a few polymorpho¬
nuclear leucocytes.
Microbiological examination :—
Nasgar tubes inoculated with spinal fluid and heart blood remain sterile;
some sporulating Gram-positive bacteria develop in haemoglobin-agar tubes.
In films from spinal fluid, after centrifugalisation, groups of diplococci are observed,
also various bacteria.
Case 59. C.W., about 30 years, black, Barbican, seen after death, at Public
Hospital Mortuary, with Dr. C. R. Edwards, of Half-Way Tree, on March 6,1913.
Patient felt ill in the evening of March 5, vomited, had some whisky, felt
better, woke up at about 1 a.m., on March 6, vomited again, had convulsions,
became unconscious, and died about 6 a.m.
Post-mortem examination , on March 6, at about 2 p.m. Well-nourished*
healthy-looking body. No skin diseases, nor parasites. No jaundice.
Lumbar puncture gives an abundant quantity of faintly cloudy fluid; about
60 c.c. are collected, but the fluid is still flowing when the needle is withdrawn-
Spinal dura, pia, and cord normal. Cerebral dura, pia, likewise normal, as well
as the brain ; lateral ventricles not distended.
Hypophysis cerebri: 13 x 6 X 3 mms., slightly hyperaemic.
Subcutaneous fat intensely yellow, orange, not icteric, but reminding of the
condition described from German Africa, as due to the ingestion of a certain kind
of vegetable oil. Visceral fat of the same colour.
Cervical , axillary , and inguinal lymph nodules considerably enlarged and
hyperaemic, the axillary mostly so.
Diaphragm at the fourth intercostal space on the right side, at the fifth on
the left.
Both pleurae show total, fibrous adhesions. The lungs show slight hyperaemia
and oedema of the postero-inferior portions. The right lung contains in its
superior and inferior lobes several fibro-calcareous noduli. The smaller bronchi
of both lungs show moderate hyperaemia and swelling of their mucosae, and some
mucous secretion. Larger bronchi, trachea, and larynx normal.
Bronchial lymph nodules slightly enlarged and anthracotic.
Thyroid body normal.
Pericardium shows total, fibrous adhesions. Heart conus, 9 x 10 X 5 cms.
Left ventricle fairly well contracted, forms apex. Pericardial fat shows
several ecchymoses on anterior surface of right ventricle. The mitral valve shows
several thickened, yellow patches on the anterior flap, and some nodular fibrous
thickening of free edge. There is no stricture nor insufficience of the valve, and
the other valves and endocard are normal. Myocard 1 cm. max. in left,
3 mm. in right ventricle ; it shows yellowish stripes and small patches ; consistence
somewhat diminished. Pulmonary artery normal. The aorta shows extensive
and intensely yellow patches of different sizes, some with superficial loss of substance,
others with calcareous plaques ; the patches make their appearance immediately
4 09
over the aortic valve, are most prominent in the arch, and are continued far
down into the abdominal portion.
The diaphragm shows over right lobe of liver a hard fibrous nodule, of about
the size of a small walnut, and close to the nodule a small, serous cyst.
Peritoneum normal. Abdominal organs normally situated.
Spleen: II X 7 X2 cms. Capsule adherent; tissue dark purple, with
prominent grey follicles ; substance soft.
Suprarenal capsules normal.
Left kidney : 11x6x3 cms. Capsule easily detached ; surface smooth,
pale grey, with patches of slight capillary hyperaemia ; cortex pale grey, hyperaemic
stripes and somewhat prominent glomeruli; columnae Bertini somewhat bulging
forwards on the cut surface, which is pale yellowish grey ; pyramids pale ; con¬
sistence considerably diminished. The right kidney shows the same size and
characteristics as the left one. Renal pelves, ureters, and bladder normal; the
bladder contains about 500 c.c. of clear, pale urine.
Ovaries contain several serous cysts, no corpora lutea, and both ligg. lata
contain also serous, multilocular cysts. Right tuba normal; left contains serous
liquid in its considerably distended fimbriate extremity.
Corpus uteri : 9 cms ; collum 5 cms., transverse diameter of corpus 8 cms.
The perimetrium is normal, except for a small serous cyst on left surface. The
endometrium cervicis appears normal, the endometrium corporis is slightly swollen
and covered with muco-purulent secretion. The posterior wall of the corpus
contains an almost globular mass, 6 cms. in diameter ; consistence soft, elastic.
Section shows a brain-like structure with some fatty masses. The anterior wall
contains a small, submucous nodule, of similar aspect and consistence as the larger
one.
Liver: 27 X 14 x6 cms. Capsule normal; surface spotted, purple and
dark yellow with smaller and larger hyperaemic patches, the whole aspect similar
to that of nutmeg liver ; consistence diminished, somewhat friable.
Bile ducts normal, patent; gall bladder normal, contains greenish bile.
Pancreas of normal size, not hyperaemic, of slightly diminished consistence.
Tongue coated with thick, white fur.
Tonsils normal.
Oesophagus normal.
Stomach contains alimentary fluid ; the mucosa is hyperaemic in the neighbour¬
hood of the lesser curvature, and in the cardiac portion, where several patches of
petechiae are seen.
Duodenum slightly hyperaemic. Jejunum and ileum normal, except for a
few patches of capillary hyperaemia. Appendix normal.
Colon shows patches of capillary hyperaemia.
No intestinal parasites .
Mesenteric lymph nodules slightly enlarged, not hyperaemic.
Urine : contains no albumen, no sugar and no casts.
Anatomical diagnoses : Pigmentatio telae adiposae. Poly-lymphadenitis acuta.
Sequelae pleuritidis duplicis. Noduli calcareo-fibrosi (tuberculosi) pulmonis dextri.
Bronchitis capillaris l.g. Sequelae pericarditidis. Endocarditis mitralis chronica
fibrosa. Metamorphosis adiposa myocardii. Aortitis atheromatosa et calcarea.
Nodulus fibrosus diaphragmatis. Splenitis acuta. Perisplenitis chronica. Nephritis
acuta parenchymatosa duplex. Cystes ovarii utriusque. Hydro salpinx dextra.
Cystis parva perimetrii. Tumor intramuralis uteri (Sarkoma) Tumor (metastaticus)
endometrii. Metamorphosis adiposa et hyperaemia hepatis. Hyperaemia et
petechiae mucosae ventriculi.
410
Histological examination :—
Lungs: Large fat drops are present in nearly all epithelial cells.
Pancreas: Most epithelial cells contain small fat granules. Extensive
necrobiosis occurs, irregularly distributed. Langerhans’s islets being affected as
well as the epithelium proper ; there is considerable hyperaemia which is likewise
irregularly distributed, in some cases apparently limited to Langerhans’s islets;
there is also some infiltrating haemorrhage.
Kidney : Sections show considerable increase of connective tissue, especially
in the pyramidal substance ; the arterial walls are fibrous and increased in thickness,
as well as Bowman’s capsules; there are patches of hyperaemia and in some places
interstitial haemorrhages. The epithelia of the convoluted tubules show in some
places granular protoplasm, but contain no vacuoles. Their nuclei are in some
places poorly stained or absent, but as a rule they are well preserved. In osmic
acid specimens small fat granules are seen in nearly all epithelial cells in the con¬
voluted tubules and also in the lining epithelia of glomeruli and Bowman’s capsules.
Numerous * mastzellen * are seen in the interstitial tissues.
Uterine tumour: The tumour shows the structure of a fibro-myoma, but of
somewhat atypical characters, being very rich in cells which show considerable
polymorphism and often large ovoid or irregularly-shaped nuclei (sarcoma).
Microbiological examination :—
Cultures on nasgar from spinal fluid remain sterile for several days, later some
Gram-positive staphylococci develop in one tube and diptheroid bacilli in another,
Gram-positive staphylococci develop in cultures of heart blood (nasgar).
No parasites in smears from heart blood.
The following cases are similiar to the preceding ones and are
therefore only given in abstract.
Case 37. G.B., 16 months, black, sister to E.B. case, Rock near Four Paths,
with Dr. E. R. C. Earle, May Pen.
Sudden onset of vomiting at 10 a.m., on February 4, 1913. The patient
is said to have had retraction of head, but no convulsions. Slight strabismus.
Temperature normal; pulse 140, small, slightly irregular. Death February 5 at
11 p.m.
Post-mortem examination :—
Spleen: Numerous subcapsular petechiae.
Kidneys: Pyramidal bases intensely hyperaemic. Consistence considerably
diminished. The left kidney shows pronounced superficial hyperaemia and a
few minute haemorrhages on the surface.
Liver: Surface pale, yellowish, and pronouncedly yellow in some parts;
in other places considerable hyperaemia. The consistence of the yellow portions
is soft, whilst it is about normal in the hyperaemic patches.
The stomach contains a considerable amount of dark, grey, alimentary fluid,
with no evidence of haemorrhage. Mucous membrane pale, of normal appearance.
The duodenum shows several hyperaemic patches. The jejunum and ileum
present likewise some hyperaemic patches, but the hyperaemia is much less pro¬
nounced than in the duodenal patches.
A few ankylostomes and a few ascarides are found in the upper portion of the
jejunum; no tricocephalus.
Anatomical diagnoses: Hyperaemia piae matris cerebralis et oedema incipiens.
Lympho-adenitis universalis. Nephritis acuta duplex. Metamorphosis adiposa
hepatis m.g. Pancreatitis parenchymatosa acuta. Hypostasis pulmonum*
Hyperaemia et ecchymoses pericardii. Metamorphosis adiposa myocardii.
Helminthiasis intestinalis.
Microbiological examination :—
Spinal fluid: Nasgar tubes: Gram-negative diplococci, which decompose
both glucose, maltose, galactose, lactose and saccharose after five days.
Case 42. C.W., years, black, Spanish Town Road, with Dr. C. R. Edwards,
Half-way Tree.
The previously healthy child woke up at 4 a.m., on February 14, 1913, and
vomited a small quantity of frothy mucus; convulsions set in, and coma, from
which she never recovered; she died at about 8 a.m., at the D.M.O.’s (Dr. Edwards’s)
office, to which she had been taken just previously ; Dr. E. obtained blood smears,
spinal fluid, and urine before death.
Post-mortem examination , at 3 p.m., on February 14, seven hours after death.
Spinal meninges and cord normal, not hyperaemic. Cerebral dura slightly
hyperaemic, but pia of perfectly normal appearance. Brain normal.
Right pleura normal; left pleura shows slight, but extensive adhesions and
numerous petechiae. Both lungs show a few small haemorrhagic infarcts, but are
otherwise normal, without hypostasis. Mucosae of smaller and larger bronchi,
and of trachea, and larynx, normal.
Both suprarenal capsules slightly enlarged, and both, especially the left, show
haemorrhagic infiltration.
Kidneys: Capsule easily detached; surface smooth, with slight capillary
hyperaemia ; cortex normal; pyramids pale.
The stomach contains a small quantity of bile-stained mucus; its mucosa is
perfectly normal, as is also that of the duodenum.
The liver measures 23 x 14 X 5 cms. Capsule normal; colour dark purple
on the surface; the tissue is grey, with slight increase of the portal connective
tissue; consistence on the whole slightly increased, but in some places friable.
Pancreas slightly hyperaemic, of normal consistence.
The lymph nodules in the hepatic hilus are very considerably enlarged, of
greenish-grey colour, and of soft elastic consistence.
Urine contains no albumin, and no casts.
Anatomical diagnoses : Poly-lymphadenitis acuta. Hyperaemia durae matris
cerebralis l.g. Sequelae pleuritidis sinistrae. Petechiae pleurae sinistrae. In-
farcti haemorrhagici pulmonis utriusque. Splenitis chronica. Epinephritis
haemorrhagica duplex. Enteritis follicularis l.g. Helminthiasis intestinalis (one
A scans lumbricoides).
Histological examination :—
Liver: Advanced fatty change, practically all cells being filled to distension
with moderately-sized fat drops. Some irregularly distributed capillary hyperaemia
and very considerable microcellular infiltration, penetrating from the periportal
tissues into the parenchyma.
Bacteriological examination :—
Spinal fluid: In nasgar tubes growth of Gram-negative diplococci which
decompose glucose, maltose and galactose after two days, and lactose and saccharose
after four days.
Case 52. J.E., 6 years, black, seen, after death, with Dr. C. R. Edwards,
February 24, 1913.
The patient is a brother to Elisabeth E. (case 51), and was taken ill suddenly
about 8 a.m., on February 22, shortly after his sister ; in fact the family ascribed
his illness to his having been frightened by seeing the sister’s convulsions. He had
several convulsions, but no vomiting. He died comatose, at about 2 a.m., on
February 23. He was seen by Dr. Croswell.
Duration of illness : eighteen hours.
Post-mortem examination :—
Axillary and cervical nodules considerably, inguinal slightly, enlarged and
hyperaemic.
The stomach contains alimentary fluid ; mucosa normal, not hyperaemic.
Anatomical diagnoses : Hyperaemia meningum. Hyperaemia glandulae sup-
rarenalis sin. et hypophysis cerebri. Hypostasis pulmonum. Splenitis acuta.
Nephritis parenchymatosa acuta duplex l.g. Metamorphosis adiposa myocardii,
hepatis, et pancreatis. Pigmentatio coli. Cadaverositas universalis.
The histological examination confirmed the anatomical diagnoses.
Bacteriological examination : No growth in nasgar tubes ; in haemoglobin-agar
tubes growth of Gram-negative bacteria.
Case 56. R.W., infant, black, seen with Dr. Gifford, at the Public Mortuary,
Kingston, on March 3, 1913.
Post-mortem examination :—
Stomach shows considerable hyperaemia and a few petechiae.
Anatomical diagnoses: Hyperaemia piae matris cerebralis. Poly-lymph¬
adenitis. Hypostasis pulmonum. Metamorphosis adiposa myocardii l.g., pan¬
creatis, et hepatis l.g. Nephritis parenchymatosa acuta duplex incipiens. Gastritis
ecchymotica l.g. Lymphadenitis mesenterialis. Helminthiasis intestinalis ( Ascaris
lumbricoides ).
The histological examination confirmed the anatomical diagnoses except that
the liver showed a rather intense fatty metamorphosis and some necrobiotic changes.
The pancreas showed, in addition to fatty change, some localized hyperaemia,
interstitial haemorrhages and fibrinous exudation ; also slight oedema of stroma ;
the islets of Langerhans were of normal aspect.
Microbiological examination :—
No growth in nasgar tubes* inoculated with spinal fluid.
Case 60. I.R., 4J years, black, Four Roads, seen, after death, on March 7,1913,
at Kingston Hospital Mortuary, with Dr. C. R. Edwards.
Previous health good. On March 6, in the evening, ‘ she complained of
abdominal pain, and later on she vomited. She went to sleep. Next morning
she woke up early, screaming; she had convulsions, became comatose, and died.’
Post-mortem examination :—
Lumbar puncture gives clear fluid, with admixture of blood. Intra-spinal
vessels intensely congested. Spinal dura and pia slightly hyperaemic. Cerebral
dura normal, pia slightly hyperaemic, especially over cerebellum. Brain and cord
normal.
Cervical, inguinal, and axillary lymph nodules enlarged and hyperaemic,
especially the latter.
Stomach normal, contains alimentary fluid. Duodenum shows considerable
follicular swelling, and the ileum shows slight swelling of solitary follicles and
Peyer’s patches in its lower part; the colon likewise shows follicular swelling;
appendix 6 cms., normal.
4 X 3
Anatomical diagnoses : Hyperaemia meningum l.g. Poly-lymphadenitis acuta.
Hypostasis pulmonum. Splenitis acuta. Nephritis acuta. Enteritis follicularis.
Metamorphosis adiposa hepatis. Petechiae pericardii visceralis.
Histological examination of liver and kidneys confirmed the anatomical
diagnoses.
Bacteriological examination :—
No growth in nasgar tubes {spinal fluid and heart blood).
Case 6i. C.W., 12 years, black, Whitehall, seen, after death, with Dr. C. R.
Edwards, at Kingston Public Hospital Mortuary, 8-III-i.
Was taken ill on March 6, with severe abdominal pain. No rise of temperature.
No vomiting. No convulsions. Died March 7, at 4 a.m.
Anatomical diagnoses : Hyperaemia meningum. Hypostasis pulmonum. Polv
lymphadenitis acuta. Hyperaemia et petechiae mucosae ventriculi. Helmin
thiasis intestinalis. Cadaverositas universalis.
Bacteriological examination: Nasgar tubes remain sterile (spinal fluid).
Group B consists of cases in which ‘ meningococci * were found in
the cerebro-spinal fluid, whilst definite inflammatory lesions of the
meninges were absent, although a more or less marked hyperaemia
was observed. Isolated cases of this nature, observed during an
epidemic of cerebro-spinal meningitis, would be accepted under this
diagnosis without much discussion. The peculiar circumstances
in our case, and more especially the close resemblance of these cases
to those of Group A, make it, however, necessary to consider very
seriously the possibility that the diplococcus infection might be
a secondary one. This question is one of the most essential of the
whole problem and will be discussed together with the general
aspects of ‘vomiting sickness.* This group includes the Cases 1,
;, 8, 12, 20, 30, 33, 43, 44, 50, 55 and 57, a total of 12 cases.
As in the case of Group A, several cases are described in full,
as types, and others only abstracted. Some of the cases are almost
identical with those given as types in Group A. The post-mortem
appearances of the meninges varied considerably in this group also,
from slight hyperaemia to intense hyperaemia accompanied by
oedema; in one case there was no hyperaemia at all and the meninges
appeared perfectly normal. Similarly, the gastric mucosa showed
in some cases normal conditions, in others hyperaemia, and in others
again, both hyperaemia and petechiae. The conditions found in
the stomach in Case 21 are particularly interesting.
Case 7. J.J.G., about 8 years, black, Chandler’s Pen. Autopsy at 3.30-5 p.m.,
about twenty-six hours after death, January 18, 1913.
The child was healthy until the foregoing day—January 17—at about 4 a.m.
when he was taken ill with vomiting, of watery fluid and food. He was conscious
to begin with, but soon became unconscious, had twitching of the limbs, vomited
a little again, and became gradually weaker, and died at 2 p.m. (he was not seen by
medical man until after his death).
Post-mortem examination (with Dr. Earle). Fairly well-nourished child.
Rigor present. Slight jaundice of sclerae.
Spinal puncture : about 20 c.c. of clear fluid.
Dura mater normal. Pia mater cerebralis shows slight diffuse hyperaemia.
Brain and spinal cord macroscopically normal.
Hypophysis cerebri appears likewise normal.
Tongue coated.
The tonsils are slightly enlarged, but show no evidence of acute inflammation.
Larynx , trachea , and bronchi contain bloody, frothy mucus, otherwise normal
Bronchial lymph nodules enlarged and anthracotic.
No adhesions but a few c.c. of clear fluid in each of the pleural cavities.
The lungs show slight hyperaemia of the lower lobules, and some oedema of
right superior lobe, but no infiltrations. No tubercular phenomena in lungs or
lymph nodules.
Thyroid body normal.
About IO c.c. of clear fluid in the pericardial cavity.
Heart: 5 X 6 X 2J cms. No pericardial haemorrhages. Myocardium
slightly pale, shows no evidence of fatty metamorphosis.
No liquid in abdominal cavity. Peritoneum appears normal.
Spleen : 11x6x2^ cms. Capsule smooth, transparent. Colour dark.
Suprarenal capsules normal.
Left kidney , 8 X 4 X ij cms. Capsule easily detached. No superficial
hyperaemia, but on section hyperaemia of pyramidal bases, which stand out very
dark on the pale cortical substance. Right kidney as left.
Renal pelves and ureters normal.
The bladder presents a normal mucosa, contains a considerable amount of
clear urine.
Liver : 21 X 11 X 5 cms. Capsule smooth ; transparent. No haemorrhages.
Consistence normal, homogeneous. Lobuli very prominent, of an intense yellowish
red colour.
The pancreas is somewhat enlarged, and is the seat of considerable hyperaemia.
Substance extremely soft.
Oesophagus normal.
The stomach contains alimentary fluid. The mucosa is hyperaemic,
especially in the vicinity of the lesser curvature. In this region numerous minute
petechiae.
The duodenum shows very slight hyperaemia, but no haemorrhages. Jejunum,
slight hyperaemia which becomes less pronounced lower down and is not at all
observed in the ileum. No enlargement of solitary follicles nor of Peyer’s patches.
Four ascarides in the small intestine, and a few tricocephali in coecum. No
ankylostomes.
Bile ducts and gall-bladder show normal mucosa ; the bladder contains dark
brownish bile.
The large bowel and vermiform appendix show normal conditions.
Anatomical diagnoses: Hyperaemia piae matris cerebralis l.g. Tonsillitis
chronica. Anthracosis lympho-bronchialis. Nephritis parenchymatosa subacuta.
4*5
Hyperaemia et petechiae mucosae ventriculi. Helminthiasis intestinalis {A sc arts
lumbricoides , Tricocephalus trickiurus).
Histological examination :—
Mesenteric lymph nodules : oedema of stroma.
Liver: Marked fatty change, evenly distributed throughout the lobules.
Hyaline and oedematous thickening of the stroma, corresponding both to portal
and to central veins.
Pancreas : Interstitial oedema.
Spleen : Oedema of coarse trabeculae.
Kidney : No oedema, but slight increase of stroma.
Microbiological examination :—
Nasgar tubes inoculated with spinal fluid show growth of Gram-negative
diplococci which change glucose and maltose but not saccharose, lactose, galactose
and mannite.
Case 12. A.M., 3 years, black, at ‘ Four Roads,’ St. Andrew (seen with
Dr. L. O. Croswell).
The girl has been subject to colds in the head, but has otherwise been healthy.
She has never passed any worms, according to her family. In the morning of
January 21, 1913, she appeared slightly unwell, and at noon-time she started
vomiting clear fluid and continued vomiting during the afternoon. She had no
febrile symptoms, but at 5 p.m. convulsions set in. Dr. Croswell saw the patient
at 5.30 p.m. ; she was then comatose and limp, showing general muscular relaxation.
There was no rigidity, no Kernig’s symptom, and no abdominal retraction. Four
or five seizures were observed, lasting for about half a minute each time, and
consisting in tonic, rather than clonic, contractions. There was seen froth at the
mouth, and a sort of hiccough, but no vomiting. She died at 6.15 p.m., apparently
from respiratory failure; the pulse could be felt, and the heart went on beating
for some time after breathing had stopped altogether.
The pulse was rather weak during the whole of the time during which the
child was under observation. Adrenalin was given subcutaneously, as a stimulant,
by Dr. Croswell.
Two years ago a brother of the deceased (then about 11 years old) had similar
symptoms, but recovered under stimulating treatment. This boy was taken
ill at a time when an epidemic outbreak of vomiting sickness reigned at 1 Four
Roads.’
Post-mortem examination , at 10 a.m., on January 22, with Dr. Edwards. Body
well nourished. No rigor. No jaundice.
Spinal fluid is taken by lumbar puncture ; it appears clear, but is mixed with
blood.
Tongue clean. Tonsils normal. Larynx , trachea , and bronchi without patho¬
logical changes, and empty, except the secondary bronchi which contain a small
quantity of frothy mucus.
No adhesions, and no fluid in the pleural cavities.
Lungs slightly hyperaemic in postero-inferior parts. No oedema, and no
infiltrations. No infarcts.
Bronchial lymph nodules not enlarged, of pinkish colour.
No anthracosis, and no tubercular phenomena in lungs and bronchial lymph
nodules.
Heart: X \\ X 2 cms. About 5 c.c. of clear fluid in pericardium.
Pulmonary artery free. Left ventricle well contracted. Myocardium pale,
\\6
greyish, without haemorrhages and yellowish patches. Consistence slightly
diminished.
Thyroid body and Thymus macroscopicaily normal.
Peritoneum of normal aspect. Position of abdominal viscera normal.
Spleen : 6 X 34 X I cms. Capsule smooth, transparent. Colour dark, with
pale, prominent follicles. Consistence normal.
Mesenteric lymph nodules considerably enlarged, of pinkish colour, with slightly
prominent vessels. Consistence normal.
Suprarenal bodies macroscopicaily normal.
Left kidney : 6 X 34 X l£ cms. Capsule easily detached. Surface smooth,
stellate veins hardly visible. Cortex pale, especially in proximal parts. Small
yellowish patches irregularly distributed in cortical substance. Pyramidal bases
slightly hyperaemic, apices pale. No haemorrhages. Consistence slightly
diminished. Right kidney : Same dimensions and aspect as left, only slightly paler.
Renal pelves , ureters , and bladder normal, the latter containing a few c.c. of
pale urine.
Oesophagus normal.
The stomach contains a small quantity of alimentary fluid. The mucosa is
slightly swollen and shows diffuse hyperaemia, mostly pronounced in the vicinity
of the lesser curvature. In this portion of the mucosa there are a few groups of
minute petechiae.
Duodenum: Slight hyperaemia.
Jejunum , ileum , and large bowel pale, without ulcers and haemorrhage. Solitary
follicles enlarged in ileum, and jejunum, and Peyer’s patches enlarged and hyper¬
aemic.
Liver : 17 x 11 X 5 cms. Capsule smooth, transparent. Colour irregular,
pinkish, with yellowish patches of different sizes, and a few hyperaemic patches.
Substance homogeneously yellowish grey; the small blood vessels hardly visible.
Consistence slightly diminished, not friable.
Pancreas: 10 cms. long. Tissue slightly hyperaemic, and of considerably
diminished consistence.
Bile ducts and gall bladder: Mucosa normal. Considerable amount of dark
greenish yellow bile in bladder.
Genital organs normal, except that both ovaries contain small serous cysts.
Meninges: Dura mater normal. Pia mater spinalis normal; pia cerebralis
slightly hyperaemic, in particular on the convex surface. No oedema and no
inflammatory exudations. Lateral ventricles not enlarged, contain a very small
quantity of clear fluid.
Brain and spinal cord macroscopicaily normal.
Anatomical diagnoses : Hyperaemia piae matris cerebri. Hypostasis pulmonum
levi gradu. Anaemia et metamorphosis parenchymatosa myocardii. Hyperaemia
oedema levi gradu et petechiae minimae mucosae ventriculi. Enteritis follicularis.
Anaemia et metamorphosis adiposa hepatis. Anaemia renum (et metamorphosis
adiposa ?). Pancreatitis parenchymatosa acuta.
Histological examination :—
Liver : Intense fatty metamorphosis and very slightly marked necrobiotic
phenomena. Marked increase of connective tissue, especially around portal vessels,
but also penetrating into the lobules, separating small groups of cells from each
other ; considerable microcellular infiltration, especially in the portal stroma.
Spleen : Intense diffuse hyperaemia ; follicles show necrobiotic changes.
Brain: No hyperaemia, no perivascular cell-infiltration.
4 '7
Kidney : Marked granular appearance and moderate vacuolization (fat) of
epithelia of convoluted tubules ; marked karyolysis in the same class of epithelia ;
glomeruli considerably retracted. Marked increase of stroma, especially in
pyramidal portions, slight microcellular infiltration.
Pancreas: Patches of beginning necrobiosis; Langerhans’ islets somewhat
prominent.
Microbiological examination :—
Heart bloody nasgar tubes remain sterile for four days. Sanguinolent spinal
fluid: Gram-negative diplococci (in the original culture the results of Gram and
Claudius staining were somewhat irregular, in subculture the results were always
negative), which change glucose, maltose, and galactose but not lactose, saccharose,*
and mannite. Spleen smears show small numbers of plump bacilli and a few
chains of diplococci.
Case 20. T.J., 12 years, coolie (Hindoo), May River, St. Mary, seen on
January 31, 1913, with Dr. Joslen, Annotto Bay. Was living in a dirty hut, with
his father, mother and two brothers, all in a half-starved condition. The father
was ill with stomatitis, vomiting and dysenteriform diarrhoea, and the mother was
dying from phthisis, with acute pneumonia. The brothers were in good health.
According to later information, the father recovered but the mother died.
Water supply from river; no latrines ; house on the side of a steep hill,
drainage consequently good.
Deceased retired to bed on the evening of January 30, apparently well. Next
morning he was found dead in his bed. (Clinical notes by Dr. Joslen).
Post-mortem examination , on January 31, at about 6 p.m. Poorly-nourished
body. Rigor present. No jaundice. No cutaneous affections, but numerous
insect bites and scratches on lower extremities.
Spinal puncture gives no fluid. Dura normal. Slight diffuse hyperaemia of
pia spinalis and cerebralis, mostly pronounced over cerebellum. No oedema, no
exudation. Lateral ventricles not dilated. Brain and cord normal.
Pleurae normal. Lungs slightly anthracotic, otherwise normal.
Bronchial lymph nodules normal.
Larynx , trachea , and bronchi are hyperaemic and contain frothy fluid.
Thyroid body pale, otherwise normal.
Pericardium normal. Endocardium and valves normal; myocardium likewise,
as well as pulmonary artery and aorta.
Peritoneum normal. Abdominal organs normally situated.
Spleen: 16 x 10 x 3 cms. Capsule normal; colour dark greyish red ;
consistence semi-fluid. Tissue uniform ; follicles greyish, not prominent.
Suprarenal capsules normal.
Left kidney: 8 X 5 X IJ cms. Capsule easily detached; surface smooth,
reddish with large yellowish patches ; stellate veins not distended ; tissue uniformly
greyish red, with yellow patches in cortex. Consistence slightly diminished ; the
right kidney presents the same characters as the left.
Liver: 20 X 15 X 6 cms. Capsule normal; colour reddish grey, with
yellow patches, and on the right lobe a number of minute haemorrhages; con¬
sistence normal.
Pancreas hyperaemic, soft.
Tongue heavily coated. Tonsils without signs of acute inflammation.
The stomach shows on the central portion of the anterior wall eight dark,
almost black, blood clots embedded in the mucous membrane ; when removed
a red spot is left, but no actual opening of a vessel is to be seen. No hyperaemia
of the mucosa and no petechiae.
Duodenum normal. The small intestine contains in many parts, especially in
the middle part of the jejunum, numerous small, half-digested blood coagula,
which are freely movable ; no haemorrhages in the mucosa, no ulcers, no follicular
swelling and no hyperaemia. Appendix and large bowel normal.
Anatomical diagnoses: Hyperaemia piae matris. Splenitis acuta. Pan¬
creatitis acuta parenchymatosa. Nephritis parenchymatosa acuta duplex l.g.
Metamorphosis adiposa hepatis l.g. Haemorrhagiae ventriculi.
Histological examination :—
Spleen : Diffuse hyperaemia.
Liver: Localized hyperaemia. Cell infiltration of portal connective tissue.
Moderate fatty change. No oedema of stroma.
Pancreas : Oedema of stroma. Necrobiotic changes of parenchyma.
Kidney : Slight oedema of stroma. Advanced necrobiotic changes of epithelia
in convoluted tubules.
Microbiological examination :—
Spinal fluid , nasgar : Gram-negative diplococci which change glucose and
maltose, but not lactose, galactose, saccharose, and mannite.
Case 44. E.P., 2 years, black, Kings Gate, seen, after death, with Dr. C. R.
Edwards, Half-way Tree.
Previous health good. Sudden onset of vomiting, followed by convulsions,
death in 3-4 hours.
Post-mortem examination , February 15, 1913. Body fairly well nourished;
rigor present; no jaundice ; on the buttocks and lower extremities numerous scars
as from scratching and small furunculi. Lice in hair, no other cutaneous parasites.
Lumbar puncture gives about 3 c.c. of clear fluid. Dura and pia spinalis show
slight hyperaemia. Dura cerebralis normal; pia shows slight hyperaemia of
convex surface ; no exudation. Brain and cord normal. No tubercular affection.
lnguinal y axillary and cervical lymph nodules moderately swollen ; some of the
inguinal and cervical nodules hyperaemic.
Diaphragm at fourth intercostal space on the right side, at the fifth on the left.
Pleurae without adhesions, contain no fluid ; the right pleura on the inferior
lobe shows a number of miliary tubercles.
Lungs slightly hyperaemic, especially in the postero-inferior portions ; left
lung otherwise normal. The right lung shows in its middle and inferior lobes
numerous peribronchial tubercular nodules.
Bronchial lymph nodules on the right side are considerably enlarged and show
caseous changes.
The smaller bronchi of the right lung show in many places caseous infiltration ;
those of the left lung, as well as the larger bronchi, trachea and larynx show
hyperaemic swelling of the mucosa and considerable mucous secretion.
Thymus .* 9 x 5 X i cms; appearance and consistence normal.
Thyroid body normal.
Pericardium contains a few drops of clear fluid. Pulmonary artery and aorta
normal. Epicard, endocard and valves normal. Heart conus 5x5x2} cms.;
left ventricle fairly well contracted ; myocard slightly pale, of normal consistence.
Peritoneum normal, without fluid ; abdominal organs normally situated.
Tongue covered with white fur, on its whole surface. Tonsils slightly enlarged ;
no signs of acute inflammation, no haemorrhages and no tubercular phenomena.
Oesophagus normal.
Stomach contains alimentary fluid ; mucosa shows follicular swelling, is other¬
wise normal.
4 i9
Duodenum shows intense follicular swelling. Jejunum, ileum and appendix
normal. Colon* shows slight follicular swelling and punctate pigmentation
corresponding to the folliculi.
The bile ducts are patent; their mucosa is normal, as well as that of the gall
bladder, which contains a moderate quantity of greenish bile.
Liver : 19 x 12 x 5 cms. Capsule normal; colour dark purple ; tissue
shows a few yellowish patches, is otherwise normal.
Pancreas of normal size and colour, but of diminished consistence.
Spleen : 9 x 4 X ij cms. Capsule normal; colour normal; consistence
soft; the tissue contains a number of miliary tubercles.
Suprarenal capsules normal.
Left kidney: 7 X 3 X 2j cms. Capsule easily detached; surface smooth,
pale, with considerable distension of stellate veins; cortex pale grey with yellowish
patches ; pyramids slightly darker, but not hyperaemic; consistence diminished.
A few miliary tubercles in the tissue. Right kidney as the left, except that it does
not contain any tubercles. Mucosa of renal pelves, ureters and bladder normal.
Genital organs normal.
Urine : faintly acid, contains a small quantity of albumen and a few granular
casts.
Histological examination :—
Cervical medulla : Moderate hyperaemia.
Spleen : Diffuse hyperaemia ; thickening of arterial walls. No histological
tuberculosis.
Myocardium : Moderate fatty change.
Lungs : Caseous and miliary tuberculosis. -
Bronchial lymph nodules : Hyperaemia ; caseous and miliary tuberculosis.
Liver : Fatty change, in some parts moderate, in others intense.
Pancreas: Localized hyperaemia, partial necrobiosis, prominent Langerhans’
islets.
Kidney : Diffuse hyperaemia, granulated protoplasma of epithelia in some
convoluted tubules and nuclei sometimes poorly stained.
Stomach: The epithelial cells are as a rule fairly well preserved; some of
them considerably swollen (hypersecretion ?). Glands are seen beneath the ordinary
level of the mucosa completely embedded in connective tissue, but apparently not
penetrating the muscularis mucosae which has been displaced, following as a
continuous layer underneath the glands. Lymphoid tissue is scarce; there is
moderate hyperaemia.
Case 50. J.M., 3 years, black, Old Hope Road, St. Andrew, seen at Kingston
Public Hospital, with Dr. Thomson, February 21, 1913.
Admitted, at 12 noon, on February 21, with a history of convulsions of sudden
onset, from early in the morning on the same day.
She is unconscious; there is no rigidity of the limbs; head not retracted;
muscles of neck somewhat rigid ; Kernig’s sign not obtained. Knee-jerks absent.
Pupils somewhat dilated, no reaction to light. A fresh, small burn is seen on the
left wrist (hot bottles had been applied at home).
Temperature 98*4. Breathing laboured. Moist rales over front of chest.
Heart’s action very rapid and«irregular. Abdomen not distended, nor retracted.
Bowels act after enema. No vomiting.
The temperature went down to 98-2 later on ; the child had several attacks of
convulsions, during which the extremities were in a state of tonic contraction, and
420
were moved slowly to and fro ; no clonic movements were observed. One attack
was observed for at least ten minutes, and lasted probably longer.
The child died at 5.30 p.m.
Post-mortem examination : February 22, at 10.30 a.m., seventeen hours after
death. Body fairly well nourished. Slight rigor. No jaundice. No skin
affection.
Cervical , axillary and inguinal lymph nodules considerably enlarged and
somewhat hyperaemic.
Spinal fluid not obtained, as skull had been opened by assistant.
Dura cerebralis and spinalis normal; pia moderately hyperaemic on convex
surface of brain ; spinal pia not hyperaemic, but shows dark pigmentation. Cord
and brain substance normal; the lateral ventricles contain a few drops of serous
fluid, from which culture is made.
Hypophysis cerebri normal.
Pleurae normal. Both lungs show slight diffuse hyperaemia, and, on the cut
surface, mucous secretion escaping from the capillary bronchi. The smaller and
larger bronchi, and trachea, and larynx are considerably hyperaemic, and contain
mucous secretion.
Bronchial lymph nodules moderately enlarged and hyperaemic. No tubercular
changes.
The pericardium contains a few c.c. of dear fluid. Heart conus 4 X 5 X i\
cms.; left ventricle somewhat flabby; epicard, endocard and valves normal.
Myocard pale, with yellowish patches, of diminished consistence. Pulmonary
artery and aorta normal.
Thymus: 9 x 3 X J cms.; of normal appearance.
Thyroid body normal.
Peritoneum normal, without fluid. Abdominal organs normally situated.
Appendix in internal position, between the small intestines.
Diaphragm at third intercostal space on right side, at fourth on left.
Mesenteric lymph nodules considerably enlarged, but not hyperaemic.
Spleen: 7x4x1 cms. Capsule normal; numerous subcapsular and
intraparenchymatous ecchymoses; tissue pale; follicles not prominent; con¬
sistence slightly diminished.
Left suprarenal capsule hyperaemic, of normal size; right normal.
Left kidney : 7 X 4J X cms. Capsule easily detached ; surface smooth ;
pale, with patches of capillary hyperaemia ; cortex pale yellowish grey and columnae
Bertini even more pronouncedly so; pyramidal bases intensely hyperaemic.
Consistence considerably diminished.
Renal pelves , ureters and bladder normal; the bladder contains a small quantity
of pale urine.
Tongue covered on the whole surface with thick, whitish fur.
Tonsils enlarged, without signs of acute inflammation.
Pharynx hyperaemic.
Oesophagus normal.
The stomach contains alimentary fluid ; mucosa normal, except a few hyper¬
aemic patches.
Duodenum normal. Jejunum shows numerous hyperaemic patches; no
follicular swelling. Ileum shows moderate swelling of Peyer’s patches, and slight
swelling of solitary follicles. All parts of colon show a large number of superficial
erosions; colon descendens shows many hyperaemic patches.
Eight ascarides in jejuno-ileum; no other parasites.
Liver : 21x11 X 4 cms. Capsule normal; surface dark purple, with large
421
yellowish patches ; tissue in some places dark, hyperaemic, but more often yellowish
grey, and in some places distinctly yellow; consistence diminished and somewhat
friable.
Pancreas of normal size, intensely hyperaemic, and of diminished consistence.
Anatomical diagnoses: Hyperaemia meningum. Laryngo-tracheo-bronchitis
catarrhalis. Bronchitis capillaris diffusa catarrhalis. Poly-lymphadenitis acuta.
Metamorphosis adiposa myocardii et hepatis. Pancreatitis parenchymatosa acuta.
Nephritis parenchymatosa acuta duplex. Hyperaemia capsulae suprarenalis sin.,
ventriculi et intestinorum. Haemorrhagiae lienis. Ileitis acuta. Helminthiasis
intestinalis.
Histological examination :—
Liver: Intense fatty change, leucocyte groups in blood vessels.
Kidney : Moderate diffuse increase of stroma. Epithelia fairly well preserved
except in a few convoluted tubules, where their protoplasm is granular and partially
destroyed and the nuclei are undergoing karyolysis. The glomeruli are well
preserved. Osmic acid preparations show small fat droplets in the basal portions
of most of the epithelia of the convoluted tubules.
Stomach : Intense localized hyperaemia of mucosa.
Pancreas: Some parts of the sections show advanced necrobiotic changes of
both the alveolar epithelia and the islets of Langerhans; in other parts all cells
are well preserved. The limit between well-preserved and necrobiotic cells is
often quite sharp, cells of the two types being seen sometimes side by side in the
same lobule or even in the same alveolus.
Microbiological examination :—
Nasgar tubes, fluid from lateral ventricles of the brain : Gram-negative
diplococci which change glucose, galactose, and maltose, but not lactose and
mannite. No growths in tubes inoculated with heart blood. No micro-organisms
in smears of peripheral blood and heart blood.
Case 57. B.M., 5 years, black, seen, after death, with Dr. J. Huntly Peck,
Spanish Town, March 5, 1913.
No clinical data obtainable.
Post-mortem examination , at 12 noon. Body fairly well nourished. No rigor.
No jaundice. No skin affection, nor parasites.
Spinal puncture gives no fluid, even though repeated directly through the
intertransversal ligaments in the lumbar and cervical regions.
Dura mater spinalis slightly hyperaemic, pia considerably hyperaemic, and
dark-pigmented, especially on the sides. Cord normal.
Dura cerebralis normal, pia considerably hyperaemic, especially on the convex
surface. Brain substance slightly hyperaemic.
Cervical , axillary and inguinal lymph nodules considerably enlarged and some
of them hyperaemic.
Pleurae normal. Lungs slightly hyperaemic in postero-inferior parts. The
smaller bronchi show slight hyperaemia of mucosa and mucous secretion ; larger
bronchi, trachea and larynx normal.
Bronchial lymph nodules slightly enlarged and hyperaemic.
Thymus .*9x4x4 cms., of normal aspect.
Thyroid body normal.
The pericardium contains a few c.c. of clear fluid. Epicardium intensely
hyperaemic, but shows no haemorrhages. Heart conus 5 x 6 x 24 cms. Left
ventricle well contracted, forms apex. Myocardium slightly pale ; endocardium
and valves normal. Pulmonary artery and aorta normal.
422
Diaphragm at fourth intercostal space on both sides.
Peritoneum normal. Abdominal organs normally situated.
Spleen : 9x6x2 cms. Capsule normal; colour dark slaty ; consistence
slightly diminished.
Suprarenal capsules normal.
Left kidney : 6J X 4 X 2 cms. Capsule easily detached, surface smooth,
pale, with slight distension of stellate veins; cortex pale greyish; pyramids
normal, consistence normal. Right kidney, same characters.
Renal pelves , ureters and bladder normal; the latter contains a considerable
quantity of pale, clear urine.
Genital organs normal.
Mesenteric lymph nodules considerably enlarged and slightly hyperaemic.
Tongue coated, on its whole surface, with thick white fur.
Tonsils normal.
Oesophagus normal.
Stomach shows patches of capillary hyperaemia.
Duodenum normal. Lower down in the intestine hyperaemic patches.
Bile ducts and gall bladder normal, patent. Bile green.
Liver : 23 x 12 x 6 cms. Capsule normal; colour uniform, greyish purple,
tissue of normal aspect.
Pancreas hyperaemic, soft.
Hypophysis cerebri normal.
Anatomical diagnoses : Hyperaemia piae matris. Poly-lymphadenitis acuta.
Bronchitis capillaris l.g. Pancreatitis parenchymatosa acuta. Hyperaemia renum l.g.
Histological examination :—
Axillary glands : Intense hyperaemia, oedema of stroma.
Spleen: Intense pigmentation.
Pancreas: Intense necrobiosis in patches, oedema of stroma.
Liver : Intense fatty change.
Kidney : Diffuse hyperaemia, more intense and with haemorrhages in some
places ; beginning necrobiosis of epithelia in convoluted tubules. Oedema of
stroma.
Microbiological examination :—
Cultures on nasgar (fluid from lateral ventricles of the brain) : Gram-negative
diplococci, small groups, which change glucose, galactose, and maltose, but not
saccharose, lactose, and mannite.
The following cases, of which only very brief notes are given,
were similar to the above types of group B.
Case i. B.D., 4 years, black, healthy from her birth until she was suddenly
taken ill, on January 13, at 4 a.m. Convulsions, but no vomiting. Death at
about about 7 a.m.
Post-mortem examination (with Dr. Earle, May Pen, at 6 p.m., January 13).
Spinal puncture gives about twenty drops of clear fluid. Diffuse hyperaemia
of pia cerebralis, mostly pronounced on some parts of the convex surface. Slight
oedema.
The stomach contains alimentary liquid without blood, and with no excess of
mucus. Considerable hyperaemia and a few minute haemorrhages in the mucosa.
Diagnoses: Hyperaemia et oedema l.g. meningum cerebralium. Lympho-
adenitis mesenterialis simplex. Hyperaemia et haemorrhagiae renum minimae.
4 2 3
Hyperaemia et petechiac mucosae ventriculi. Enteritis follicularis Lg. Helmin¬
thiasis intestinalis (Ascaris lumbricoides). Metamorphosis adiposa hepatis l.g.
Histological examination :—
Lymph nodules : Slight increase of stroma.
Liver: Patches of hyperaemia are seen in various parts of the sections ; the
stroma around the portal vessels and the capsule are slightly increased in thickness
and there is a beginning microcellular infiltration. The parenchyma is well
preserved.
Kidney: Patches of hyperaemia are seen in various parts of the sections, and
the stroma is slightly increased.
Suprarenal capsules: Considerable hyperaemia and marked vacuolization of
cells.
Microbiological examination :—
Nasgar tubes inoculated with spinal fluid show growth of Gram-negative
diplococci which change glucose, maltose and galactose, but not saccharose and
lactose when examined in Jamaica, but in Liverpool they give different reactions
which will be mentioned later on in the general discussion.
Smears from brain , spleen , and liver contain various bacilli and cocci.
Case 8. H.S., 3J years, black, Post Road, by Chapelton. (Seen after death
with Dr. Thomson). The child was, according to the mother, healthy until
January 18, 1913, at 5 a.m., when she started vomiting and continued vomiting
until death took place two hours later. No convulsions. The previous day she
had eaten some unripe yam ; the mother partook of the same and had some vomiting
immediately afterwards.
Post-mortem examination :—
The stomach contains alimentary fluid and a small amount of mucus. No
haemorrhage; no hyperaemia.
Duodenum : Slightly hyperaemic.
Diagnoses: Hyperaemia piae matris cerebralis l.g. Hypostasis pulmonum l.g.
Enteritis follicularis l.g. Helminthiasis intestinalis (Ascaris lumbricoides).
Lymphadenitis mesenterialis.
Microbiological examination :—
Nasgar tubes inoculated with heart blood remain sterile; tubes inoculated with
the sanguinolent spinal fluid show growth of Gram-negative diplococci which change
dextrose, maltose, and to a very slight degree galactose, but not lactose, saccharose, and
mannite.
Case 30. D.B., 9 months, black, Kingston Public Hospital, Dr. Thomson,
January 31—February 1, 1913.
Sudden onset of vomiting and convulsions ; quite unconscious when admitted;
limbs rigid ; head retracted ; Kemig’s sign marked in both legs, left more than right.
No fluid obtained by lumbar puncture. Given 50 millions meningococcus-vaccine.
February 1. Rested fairly well during night. Only two attacks of convulsions
no vomiting ; limbs still rigid. The child appears conscious. Temperature normal.
Slight nystagmus. Slight internal strabismus, of left eye.
Post-mortem examination (performed by Dr. Thomson, who preserved the
organs for me, as I was absent in the country).
Meninges congested. Lateral ventricles distended with blood-stained fluid.
Upper part of spinal pia dark pigmented..
Internal organs normal.
Anatomical diagnosis : Hyperaemia meningum.
4 2 4
Bacteriological examination :—
Nasgar tubes inoculated with spinal fluid show growth of Gram-negative diplo-
cocci which change glucose, maltose and galactose, but not saccharose, lactose and
mannite.
Case 33. E.B., 2 years, black. Rock, near Four Paths, with Dr. E. R. C. Earle.
Has been slightly ill for about three months. On February 3 he was unable
to get up, * he could not drink, became stiff and died at II a.m.’ Vomiting not
mentioned, nor convulsions.
Post-mortem examination : No jaundice.
Lumbar puncture gives clear, translucent fluid. Spinal cord not examined.
Dura mater cerebralis normal; pia shows moderate hyperaemia, mostly pronounced
on the posterior portion of the convex surface ; no hyperaemia on the basis of the
brain, but a small collection of slightly turbid fluid. The lateral ventricles are not
dilated. The brain substance is of normal appearance.
Anatomical diagnoses: Hyperaemia meningum et mucosae ventriculi. Petechiae
pericardii. Anaemia organorum. Helminthiasis intestinalis. Tuberculosis caseosa
nodulorum lymphaticorum bronchialium.
Bacteriological examination :—
Spinal fluid , nasgar tubes : Gram-negative diplococci which change glucose,
maltose and galactose, but not lactose, saccharose and mannite.
Case 43. C.B., 2 years, black, Old Hope, The Quarry, with Dr. C. R. Edwards,
Half-Way Tree.
The child seems to have had slight cold for about two hours, then became very
ill, vomited and died in convulsions at 5 a.m., on February 14, 1913.
Post-mortem examination , at 3 p.m., on February 14, about ten hours after death.
Body fairly well nourished. No jaundice.
Lymph nodules: Axillary slightly enlarged, moderately hyperaemic ; inguinal
considerably enlarged, slightly hyperaemic; cervical moderately enlarged and
hyperaemic.
Spinal puncture gives about 5 c.c. of clear, afterwards slightly turbid fluid. Spinal
and cerebral durae normal, piae slightly hyperaemic, particularly on the convei
surface of the brain'. No exudation. Brain and cord normal. Lateral ventricles
not distended.
Mesenteric lymph nodules are very much enlarged, and show punctate hyperaemia.
Spleen : 8x4x2 cms. Capsule normal, tissue normal, pink.
Anatomical diagnoses: Poly-lymphadenitis acuta l.g. Lymphadenitis mesen-
terialis m.g. Hyperaemia meningum. Sequelae pleuritidis sinistrae. Infarctus
haemorrhagicus pulmonis dextri. Bronchitis. Enteritis l.g. Hyperaemia mucosae
ventriculi
Histological examination :—
Axillary lymph nodules: Intense hyperaemia and infiltrating haemorrhages.
Mesenteric lymph nodules: Moderate hyperaemia.
Kidney : Irregularly distributed patches of hyperaemia.
Pancreas : Oedema of stroma.
Microbiological examination :—
Nasgar tubes of spinal fluid: Gram-negative diplococci and Gram-negative
cocco-bacilli. The diplococci are isolated and are found to change glucose and
maltose, but not galactose, lactose and mannite. Also the examination of the
centrifugalized deposit shows Gram-negative diplococci and several other bacteria.
425
Case 55. A.T., 6 years, black, Barbican, St. Andrew, sent to hospital by
Dr. Crosswell, Public Hospital, Kingston, March 2, 1913.
The child started vomiting on the evening of February 28, 1913, has been
vomiting since then, the vomited substance being yellow. Convulsions began at
7.30 p.m. on the following day. When seen by Dr. Crosswell, all limbs were relaxed,
there was no retraction of head, nor of abdominal wall; Kernig’s symptom ; pupils
sluggish, equal; the child was semi-comatose, but could be roused, and could sit up ;
asked for water. He was immediately sent to hospital. On his arrival at the hospital
he was moribund ; the limbs were jerking, the head drawn back ; temperature sub¬
normal ; skin cold and clammy; pupils dilated.
He was admitted at 8.30 p.m., and died at 9 p.m., on March 2.
Post-mortem examination, , at 11 a.m., on March 3.
Lumbar puncture gives about 10 c.c. of clear fluid. Spinal meninges normal,
without any hyperaemia; cord normal. Cerebral dura slightly hyperaemic; pia
shows intense hyperaemia and some oedema on convex surface, slight hyperaemia on
cerebellum, no hyperaemia at the basis. The lateral ventricles contain a small
amount of slightly turbid fluid. The hemispheres show slight hyperaemia of the
brain substance which is otherwise normal.
The cervical and inguinal lymph nodules are considerably enlarged and slightly
hyperaemic, the axillary nodules considerably enlarged and intensely hyperaemic.
The pericardium contains a few drops of clear fluid ; the epicard shows numerous
petechiae and small ecchymoses on both the posterior and anterior surfaces of left
ventricle, and to a less degree on the anterior surface of the right ventricle.
The stomach shows considerable capillary hyperaemia, corresponding to the
whole length of the lesser curvature and neighbouring portions, and a few petechiae
in cardiac portion; slight follicular swelling.
Duodenum slightly hyperaemic. Jejunum shows patches of capillary hyperaemia ;
no follicular swelling. Ileum shows hyperaemia and swelling of some of the lower
Peyer’s patches and considerable swelling of the solitary follicles in the last portion.
Appendix 6 cms., normal. Colon normal. No parasites.
Mesenteric lymph nodules are slightly enlarged, pale, of normal consistence.
! Tonsils: There is a superficial necrosis, diffuse microcellular infiltration into
the connective and muscular tissues ; the vessels are enormously distended.
Urine : Yellow, slightly cloudy, acid, contains no albumen, but gives a faint
reaction for nucleo-albumen, it contains no sugar, no bile pigment, and no casts.
Anatomical diagnoses: Hyperaemia meningum et cerebri. Lymphadenitis
multiplex. Hyperaemia et oedema pulmonum. Metamorphosis adiposa hepatis et
pancreatis. Nephritis parenchymatosa acuta duplex. Ecchymoses epicardii et
lienis. Hyperaemia ventriculi c. petechiis. Enteritis follicularis.
Histological examination :—
Axillary lymph nodules: There is moderate diffuse hyperaemia and some peri¬
vascular blood-infiltration.
Myocard: Sections of fragments fixed in Flemming’s fluid show minute fat
droplets in the muscle fibres.
Spleen: Diffuse hyperaemia.
Suprarenal capsule : Intense diffuse hyperaemia of the organ and of the surround-
ng connective and fatty tissues.
Kidney : Irregularly distributed patches of intense hyperaemia ; the cells of the
convoluted tubules are swollen and granular, with vacuoles near the basal membranes.
There is a slight increase of the stroma, accompanied by a slight microcellular infiltra¬
tion.
426
Liver: Considerable capillary hyperaemia, slight microcellular infiltration
of stroma and penetrating amongst the liver cells, which show advanced fatty
change mostly marked in the portal and intermedial zones.
Stomach : The sections show the transition from the oesophagus to the stomach.
The oesophageal epithelium is fairly well preserved. The gastric epithelium on the
contrary is to a very large extent destroyed and in some place completely destroyed.
There is an intense hyperaemia, and in the gastric mucosa also minute haemorrhages.
The gastric lymphoid follicles are enlarged and contain numerous polymorphonuclear
leucocytes and there is also diffuse microcellular infiltration of both mucosa and
submucosa of the stomach, whilst the same phenomena are but slightly pronounced in
the oesophagus. There is some oedema of the stroma. Numerous 4 mast-zellen ’ in
all coats, especially of the oesophagus. Bacteria are exceedingly numerous in the
superficial portions of the stomach, and in some places groups of bacteria are observed
in the deeper layers in blood vessels.
Centrifugalization-deposit of the spinal fluid shows numerous endothelial cells,
few lymphocytes and a few polymorphonuclear leucocytes.
Microbiological examination :—
Spinal fluid , cultures on nasgar: Gram-negative diplococci which change
glucose, maltose and galactose, but not saccharose, lactose and mannite. No
micro-organisms are seen in smears from spinal fluid and blood from lateral ventricles.
Group C contains cases resembling those in the two preceding
groups in their clinical and anatomical characteristics, but no
bacteriological examination of the cerebro-spinal fluid has taken
place. In fact, the cases in the three groups, A, B and C, are often
so much alike that the only constant difference is that in B
meningococci have been demonstrated, in C they have not been
examined for, and in A they have not been found, though carefully
examined for. A satisfactory classification of the cases in this
group cannot be attempted until the Groups A and B have been
discussed. Group C contains 11 cases, namely, Cases 2, 5, 10, ii,
13, 18, 24, 25, 27, 32 and 48.
The cases in this group are given in abstracts only.
Case 2. R.B., 4 years, coloured (seen with Dr. Earle, May Pen), January 13.
Lives in the house next door to the one in which Case I died, and was reported
ill whilst the autopsy on this case was being performed. The child started
vomiting at 7 p.m., and was seen at 9 p.m., when it was in a state of slight
collapse, with soft and somewhat irregular pulse, about 90. It was stimulated with
ether, and improved rapidly. Recovery.
This case was, according to Dr. Earle, one of so-called 4 vomiting sickness.’
Case 5. A.A.J., 23 months, black, male, not seen by physician before death.
Melrose.
Taken ill suddenly on January 15, at 5 a.m., with pain in stomach, and efforts
of vomiting, but no actual vomiting. No convulsions. Death took place at
7 a.m.
Post-morUm examination , with Dr. Cooke, Mandeville.
4*7
The kidney measures 6 x 3^ X i£ cms. Capsule easily detached. Surface
smooth, with minute haemorrhages and considerable hyperaemia. Renal substance
reddish grey, almost uniform throughout cortex and pyramids. A few minute
haemorrhages and yellowish patches in the substance.
Liver: 16 X 11 x 4 cms. Capsule smooth and transparent. Substance
uniformly greyish red, of normal consistence. No yellow, or soft patches.
Pancreas of normal size, colour, and consistence.
Oesophagus , stomach , and duodenum without pathological changes. No
hyperaemia except very slightly in the duodenum. The stomach contains a
small quantity of liquid food ; no blood.
Jejunum of normal characters, except for a slight enlargement of the solitary
follicles in its upper part. A few of the last Peyer’6 patches in the ileum are slightly
enlarged.
Appendix , coecum , and colon normal. No intestinal parasites.
Mesenteric lymph nodules not enlarged.
Diagnoses : Hyperaemia meningum l.g. Tonsillitis chronica. Nephritis acuta
l.g. Enteritis follicularis l.g. Splenitis acuta l.g.
Case 10. S.S., 4 years, brown, 4 Chancery Lane, seen with Drs. Dryden
and Thomson in the Public Hospital, Kingston. January 21, 1913.
Admitted with her playmate, Case 9 (group A). Symptoms and course similar,
but weakness less pronounced.
Microscopical examination of the blood shows no parasites.
Case ii. A.W., 2 months, black, 82 Ruin Lane, seen in Public Hospital,
Kingston, with Dr. Thomson. Admitted January 21, 1913.
Admitted to hospital on account of continuous vomiting and collapse; has
been ailing for one day. Breast-fed. No convulsions, and no other meningeal
symptoms. Temperature ioo° F. in rectum. Gums swollen and tender.
The following days no vomiting, and no other symptoms. Temperature
normal.
Microscopical examination of the blood shows no parasites.
Case 13. E.S., 16 years, black, Four Roads (seen with Dr. Croswell). The
patient was well until the morning of January 23, when she was taken ill, feeling
unwell and vomiting twice during the day. She was not feverish. The following
morning she felt very weak, and was taken to Dr. C’s surgery where she vomited
once. This vomit was watery, frothy, but those of the previous day were described
as ‘ bilious.’ The temperature was normal, the pulse somewhat weak. Dr. C.
called me to see the patient, and I saw her with Dr. C. at noon. She looked
normal, slightly drowsy. Pulse 92, slightly soft and irregular. No jaundice.
Heart, liver and spleen-dullness normal. No abdominal pain, tenderness or
contraction. No stiffness of neck or other muscles. No Kernig’s symptom.
No deviation of eyes. Numerous rhagadae on the lips, and a herpes-like eruption
on the chin.
No parasites were found in blood smears.
(This case was considered by Dr. Croswell to correspond fairly well to the
picture of 4 vomiting sickness ’).
In the following case there were several attacks of ' meningismus/
during the course of a suppurative infection. When the child died,
the possibility of a purulent meningitis was therefore considered,
but both the post-mortem findings and the final part of the illness
were in accordance with the type of vomiting sickness.
Case 18. A.B., 4 months, black, Bloxburgh, St. Andrew. Seen at Kingston
Public Hospital, with Dr. C. A. A. Thomson. January 4—29, 1913.
January 4, admitted, with abscess of axilla, observed three weeks previously ;
is said to have had convulsions recently. No vomiting, nor diarrhoea. Abscess
incised. Temperature normal.
January 14. Has had several attacks of convulsions. No rigidity ; no Kernig’s
symptom. Bowels free. Incision wound healing.
January 17. No convulsions for the past 48 hours. Wound healed, but
there is some hard infiltration of the surrounding parts. Temperature normal.
January 29. Return of convulsions yesterday, one lasting several minutes
followed by exhaustion and death.
Clinical diagnosis: Posterior basal meningitis.
Post-mortem examination: (29 January, 1913).
Anatomical diagnoses: Atrophia universalis. Anaemia organorum. Hyper-
aemia piae matris cerebralis. Pediculosis capitis.*
Case 24. M.E., Craigmill, black, adult, seen with Dr. George.
The patient had complained of cardialgia and nausea for several days. On
January 29, she started vomiting at about 2 p.m.; she had no fever. She went
on vomiting for two days, bringing up considerable, according to description
enormous, quantities of watery, yellow or greenish fluid. Temperature normal.
Pulse 92, regular, weak. No meningeal symptoms.
February 1. Patient admitted to hospital. No vomiting. Pulse and
general condition improved, after tinct. strophanti.
Case 25. R.M., 10 years, black, Craigmill. Seen on January 31, 1913, with
Dr. George.
Patient has vomited four times since morning; was seen by Dr. George at
4 p.m. There is some epigastric pain and tenderness, when he is seen by Dr. G.
and myself at 11.30 p.m. Pulse fairly good, but not strong. No vomiting since
this afternoon. No meningeal symptoms.
February 1. Improved.
Recovered.
Case 32. E.B., 6 years, black, Rock near Four Paths, with Dr. Earle. ‘ Quite
well on the morning of February 3. In the afternoon she vomited greenish
substance and went to bed. About midnight she again started vomiting and
continued till daybreak of the 4th. The jaw was locked, she could not swallow,
became stiff and ground her teeth.’
The patient died at 10 a.m., on February 4, 1913.
Post-mortem examination :—
Liver: 18 x 10 x 4 cms. Capsule normal; surface and substance uniformly
reddish grey.
The stomach contains clear alimentary fluid; the mucosa shows, on both
sides of the lesser curvature capillary hyperaemia and numerous petechiae.
429
The jejunum and ileum contain numerous ascarides; no other intestinal
parasites. The Peyer’s patches in the lower part of ileum are slightly enlarged
and hyperaemic.
The pancreas is normal.
Anatomical diagnoses: Hyperaemia piae matris cerebralis. Hyperaemia et
petechiae ventriculi. Enteritis l.g. Nephritis parenchymatosa acuta l.g. Helmin¬
thiasis intestinalis.
Case 48. L.W., 12 years, black, Windward Road, Doncaster Pen. Seen at
Public Hospital, Kingston, with Dr. Thomson.
Admitted, on February 18, 1913, with history of sudden onset of vomiting
the previous night. An aunt (case 47) and her infant were similarly attacked, the
infant dying at home.®
Temperature on admission normal, and remained so. Patient drowsy. No
vomiting. No muscular rigidity ; no Kernig’s symptom.
February 19, rested well last night. No malarial parasites in blood.
Patient recovered, and left Hospital on February 21 ; he was then apparently
quite well, but the pulse was very frequent, 100—120, weak and slightly irregular.
Case 51. E.E.E., 20 years, black, seen with Dr. Croswell, on February 23 ;
post-mortem examination with Dr. Edwards on February 24.
The patient was taken ill suddenly, on February 22, at 7 a.m. with convulsions,
was seen by Dr. C. in the afternoon, was then very violent; after chloral and
bromide she seemed to improve, though she had repeated convulsions. The
following day, just after noon, she became unconscious, and comatose, and
was in a dying condition, when seen on February 23, at 4 p.m. by Dr. C. and
myself. Temperature was then 105° F., pulse could not be felt; she reacted a
little, when given injections of ether and digitalin, so that the pulse could just be
felt, filiform, at the radials, but not counted; the heart beats were weak, about
150 to 180. She died in coma at about 5 p.m.
When seen just before death, the pupils were of normal size ; there was no
retraction of the head, but slight rigidity of the neck-muscles. No Kernig’s sign.
Abdomen natural, no enlargement of liver, or spleen. Duration of illness: 34
hours.
Post-mortem examination , at 12 noon, on February 24, about 19 hours after
death.
Body well nourished, healthy looking. Excoriations on the lips. Rigor
present. No jaundice.
Cervical , inguinal and particularly the axillary lymph nodules considerably
enlarged and intensely hyperaemic.
No spinal fluid obtained by lumbar puncture . Cerebral dura and pia without
hyperaemia and exudation ; lateral ventricles not distended ; brain substance
normal. Spinal dura, pia and cord perfectly normal in dorsal portion, the only
part examined.
Hypophysis cerebri: Normal.
Pleurae : Left pleura shows numerous small ecchymoses, especially on the
inferior lobe.
The pericardium contains a few c.c. of clear fluid; the epicard shows diffuse
hyperaemia and numerous small ecchymoses on the posterior surface of the left
* A post-mortem examination was made on the infant by Dr. Gifford who informed me
that he had found no hyperaemia of meninges and no pronounced lesions of any organ.
430
ventricle and auricle. Heart conus 9 x 10 x 5 cms.; apex is formed by left
ventricle which is somewhat flabby. Endocard and valves normal. Myocard
pale, with yellowish patches, of rather soft consistence.
Mesenteric lymph nodules slightly enlarged and hyperaemic.
Spleen: 10 x 8 X 2 cms. Capsule slightly thickened, whitish; colour of
tissue normal; follicles not prominent, but a considerable number of small (from
miliar to lenticular) yellow nodules are seen in the pulpa. Consistence normal.
Suprarenal capsules normal.
Left kidney: 12 x 6 x 3 cms. Capsule slightly adherent; surface smooth,
except along some entering vessels, where capsule is adherent; surface pale
yellowish, with patches of capillary hyperaemia ; cortex pale, yellowish, with
prominent glomeruli and some hyperaemic stripes; columnae Bertini still more
yellow and rather soft; their tissue is bulging forward on the cut surface; the
pyramids show slight hyperaemia of their bases.
Right kidney .-12x7x3 cms. Shows same characters as left.
Renal pelves, ureters and bladder hyperaemic. The calyces, pelvis and ureter
on the right side contain a yellowish, purulent fluid. The bladder contains about
100 c.c. of pale, slightly turbid urine.
Both ovaries are hyperaemic; the right contains a fresh corpus luteum and
the left also one, apparently of recent date. Both contain, besides, small serous
cysts. The Fallopian tubes are not distended ; only the left is patent; its mucosa
is apparently normal, but the ovarian extremity contains purulent fluid. Uterus
hyperaemic ; its cavity measures, from orificium internum to fundus 4 cms.;
collum 3 cms. Endometrium corporis hyperaemic and covered with slight purulent
secretion. The cervix contains muco-pus; its mucosa is swollen, with minute
haemorrhages. The portio vaginalis is discoloured, dark, with erosions.
Liver : 30 X 16 X 8 cms. Capsule normal; surface spotted red and yellowish
grey, with subcapsular hyperaemia and minute haemorrhages ; the tissue is
uniformly yellowish grey; structure enhanced by slight hyperaemia of portal
veins; consistence soft and extremely friable.
Pancreas of normal size, considerably hyperaemic and very soft.
Tonsils show small purulent particles on the surface, not enlarged.
Oesophagus: Slightly hyperaemic, contains greenish black fluid.
Stomach considerably distended, contains greenish black fluid with small black
particles; no smell of phosphorus. The mucosa shows considerable hyperaemia
of the cardiac portion, on both anterior and posterior walls, and a large number
of petechiae ; on the remaining parts of the mucosa a few small erosions are seen.
Duodenum shows hyperaemia and some petechiae.
Appendix 17 cms., normal.
Bile ducts and gall bladder normal; the former are patent, and the latter
contains a moderate quantity of greenish bile.
Urine : Turbid ; acid, contains about I per mille albumen and a few granular
casts, no indican, no bile pigments and no sugar.
Anatomical diagnoses: Poly-lymphadenitis acuta. Haemorrhagiae pleurae
sinistrae, pericardii, et capsulae hepatis. Hypostasis pulmonum. Laryngo-tracheo-
bronchitis catarrhalis. Struma simplex l.g. Metamorphosis adiposa myocardii
et hepatis m.g. Pancreatitis parenchymatosa acuta. Nephritis parenchymatosa
acuta duplex m.g. (in chronica). Pyelo-ureteritis dextra. Endometritis. Sal¬
pingitis. Hyperaemia ovariorum. Noduli lienis. Gastritis ecchymotica cum
erosionibus. Duodenitis ecchymotica. Hyperaemia jejuni et coli. Pharyngitis
acuta.
Histological examination :—
Axillary lymph nodule: Intense hyperaemia and haemorrhages, both arteries,
veins and capillaries being enormously distended; there is diffuse erythrocyte
infiltration of the tissues especially in the central portion of the nodule.
Liver : Intense fatty change and slight necrobiotic changes equally distributed
throughout the lobuli; several patches of capillary hyperaemia.
Pancreas : In the sublimate-alcohol specimens, only slight localized necrobiotic
changes are observed, but in the Flemming specimens there is marked diffuse fatty
metamorphosis present, all or nearly all cells containing minute fat droplets.
Kidney : The glomeruli are distended and some of them hyperaemic. Most
of the epithelial cells in the convoluted tubules contain large vacuoles (fat,
Flemming). A few tubules show marked necrobiotic changes of the cells. The
vacuoles, when not filling the cell entirely, always occupy the basal portion
of the cell; as a rule the cell contains only one or a few fat drops. In some
portions of the kidney the necrobiotic changes are more marked and the fatty
metamorphosis less marked. In these portions there is some capillary hyperaemia.
There is a slight increase of the connective tissue throughout the sections; there
is no microcellular infiltration.
Microbiological examination :—
No growth in nasgar and haemoglobin agar tubes inoculated with heart blood,
except some colonies of large bacteria, probably accidental infection. No parasites
in smears of peripheral blood and heart blood. No gonococci in smears from the
right renal pelvis and endometrium.
In group D we include the cases in which the usual characteristics
of meningococcus-meningitis were found, namely definite inflam¬
matory changes in the meninges and meningococci in the cerebro¬
spinal fluid. The following five cases fulfil these requirements :
4, 19, 29, 31 and 39. All these cases are given in abstract only.
Case 4. V.C., 2 years, female, black, of Craigmill, Portland, seen, after death,
with Dr. H. J. George, of Buff Bay.
The child was taken ill at 5 p.m. on January 13, having been in good health
previously. She had never before suffered from convulsions, fever, or vomiting.
She started vomiting and continued vomiting, apparently without any effort,
until her death which took place at about 5.30 p.m., i.e., about half an hour from
the onset. The child did not appear feverish, there were no convulsions, and no
stiffness of neck or other muscles was noticed.
Post-mortem examination was made at 6 p.m., on January 14, i.e., 25 hours
p.m. Well nourished child. No jaundice.
Spinal puncture gives about twenty drops of clear, yellowish fluid.
Meninges: Intense congestion of both cerebral and spinal dura and pia, but
mostly pronounced in the pia on the convex surface of the brain, and on the upper
surface of cerebellum. Slight oedema of pia. Beginning sero-fibrinous exudation
in median and Sylvian fissures.
Brain and spinal cord show, in some parts, irregularly distributed slight hyper¬
aemia, but no other macroscopical lesions.
Mouth , pharynx , tongue , tonsils , larynx , trachea and bronchi , thyroid body and
thymus are macroscopically normal. No liquid and no adhesions in pleurae.
Oesophagus , stomach and duodenum without pathological changes.
43 2
Liver: 17 x 10 x 4 cms. Capsule smooth, transparent. Colour slightly
irregular, greyish with red patches. No yellow colouring except a few small
patches in the left lobule. Substance slightly pale, otherwise normal.
Pancreas: Of normal size, colour and consistence.
Anatomical Diagnoses: Hyperaemia diffusa meningum. Leptomeningitis
cerebralis fibrinosa incipiens. Hypostasis pulmonum l.g. Infarcti haemorrhagici
pulmonis sinistri. Anaemia myocardii. Anthracosis lymphonodulorum bronchi-
alium (et tuberculosis ?) Cystis haemorrhagicus ovarii dextri. Lympho-adenitis
mesenterialis simplex. Enteritis follicularis l.g. Helminthiasis intestinalis (Ascaris
lumbricoides).
Microbiological examination :—
Nasgar tubes, spinal fluid: Gram-negative diplococci which change glucose
and maltose, but not galactose, lactose and saccharose.
Case 19. A.H., 4 years, black, 12 Beeston Street, Kingston. At Kingston
Public Hospital (Dr. Thomson), January 18, 1913
Apparently well till January 17, at 11 p.m., when he started vomiting; he
had convulsions, was stiff at times; the head was drawn back.
He was admitted to hospital early the following morning, and died at 8.30
a.m., on January 18.
Kernig’s sign was present on both sides.
Lumbar puncture gave spinal fluid under considerable pressure ; the fluid
was clear.
The urine was clear, contained no albumin, and no casts.
Post-mortem diagnoses (Dr. Scott) : Hyperaemia piae matris. Meningitis
sero-fibrinosa. Sequelae pleuritidis sinistrae. Lymphadenitis mesenterialis.
Metamorphosis adiposa hepatis, l.g.
Microbiological examination :—
Nasgar tubes, spinal fluid: Gram-negative diplococci which change glucose,
maltose and galactose, but not lactose, saccharose, and mannite.
Case 29. B.W., 12 years, male, black, admitted to Kingston Hospital,
January 25, 1913.
History of ‘ fever *; delirium on admission; temperature ioo° F.; no
retraction ; no rigidity ; no Kernig ; spleen not felt; passes urine in bed ; illness
said to have lasted a week or longer. Diagnosis of malaria ; blood shows crescents.
January 26 :—temperature ioi° F., still unconscious, had two attacks of convulsions
during night. Died January 28, 9.20 a.m.
Post-mortem examination was made at 2 p.m. (by Dr. Scott), on January 28,
i.e., 5 hours p.m.
Thoracic organs : Nothing abnormal.
Spleen is dark, greyish-black, not much enlarged. 5x3x1 cms.
Liver is dark, greyish-black.
Congestion of Pi a and surface of Brain is very marked ; thickish lymph over
cortex, with purulent patches between hemispheres; at base, greenish-yellow
pus and abundant turbid fluid. Ventricles distended, about 5—6 ozs. of slightly
turbid fluid in each. Spinal fluid turbid and abundant.
Anatomical Diagnoses : Leptomeningitis, fibrino-purulenta. Splenitis acuta
Hepatitis parenchymatosa.
Microbiological examination :—
Spinal fluid : Smears show numerous intracellular diplococci; cultures on
nasgar show colonies of Gram-negative diplococci which change glucose, maltose
and galactose, but not lactose, saccharose and mannite.
433
Case 31. H.M., 19 months, black. At Kingston Public Hospital, February
3 — 5 > I 9 I 3 *
Admitted unconscious, on February 3, at 3.45 p.m. History of vomiting and
convulsions of sudden onset.
Temperature 98. Slight strabismus. Limbs rigid. Lumbar puncture : no
fluid. February 4, Temperature 102. Has vomited once. Head retracted.
Kernig’s sign doubtful. Knee-jerks not obtained.
February 5, very weak. Cheyne-Stokes respiration. Death.
Post-mortem examination (performed by Dr. Scott, who preserved the organs
for me, as I was out in the country).
No congestion of meninges and brain, but lateral ventricles distended with
turbid fluid. Inflammatory lymph at basis of brain. Spinal canal shut off by a
layer of lymph like a diaphragm (neg. res. of lumbar punct. !) Internal organs
normal.
Anatomical Diagnosis : Meningitis.
Microbiological examination :—
Nasgar tubes, spinal fluid : Gram-negative diplococci which change glucose,
maltose and galactose, but not saccharose, lactose and mannite.
Case 39. U.M., 8 years, black, Osborne Store, Four Paths, Clarendon, with
Dr. E. R. C. Earle, May Pen.
The child was healthy until Sunday, February 9, 1913, when he was taken ill
with vomiting, at about 9 a.m. He improved after mint-tea and whisky, but on
the following night vomiting started again, at about 3.30 a.m. on February 10.
He vomited repeatedly during the day. He was seen by Dr. Earle at about 5 p.m.,
but was then already in a dying condition. He died at 6 p.m.
Post-mortem examination :—
Cervical , axillary and inguinal lymph nodules slightly swollen.
Lumbar puncture gives clear fluid. Dura mater normal. Pia mater slightly
hyperaemic on the convex surface of the brain, more pronouncedly so on the basis
and especially on the cerebellum ; it appears normal on the upper part of the
spinal cord, but there is a slight hyperaemia of its lumbar portion. There is a
small amount of slightly clouded, serous exudation at the basis of the brain, of
which liquid smears are taken. Lateral ventricles of normal aspect, as well as the
substance of brain and cord.
The stomach is somewhat dilated and contains a considerable quantity of fluid
of the character of intestinal contents. The gastric mucosa is normal in its larger
part, but shows in its central portion, on the anterior surface and close to the lesser
curvature, a large hyperaemic patch with several petechiae and a few petechiae
scattered about on the neighbouring parts. Near the pylorus, on the posterior
wall, considerable follicular swelling is observed. The duodenum shows hyper¬
aemia, ecchymoses and follicular swelling. The jejunum is hyperaemic, especially
in its upper portion ; no haemorrhages. The intestinal tract is otherwise normal ;
it contains no parasites.
Anatomical diagnoses: Leptomeningitis incipiens. Lymphadenitis simplex
l.g. Hypostasis et oedema l.g. pulmonum. Bronchitis. Metamorphosis adiposa
myocardii et hepatis. Gastritis ecchymotica et follicularis. Enteritis l.g. Splenitis
et perisplenitis chronica. Pancreatitis parenchymatosa acuta. Nephritis parenchy-
matosa acuta l.g.
Urine : Faint albumin-reaction and hyaline casts.
Histological examination :—
Spleen : Oedema of stroma.
434
Myocard: Fragmentation.
Pancreas : Extensive necrobiotic changes, most nuclei remaining unstained,
and the cell limits seldom being recognisable.
Stomach: The mucosa shows considerable hyperaemia and some superficial
haemorrhages, and diffuse microcellular infiltration.
Smears of spinal fluid after centrifugalization show a number of lympho¬
cytes and a few polymorphonuclear leucocytes.
Microbiological examination :—
Nasgar tubes, sfinal fluid : Gram-negative diplococci which change glucose,
maltose and galactose, but not lactose, saccharose and mannite. Same result on
haemoglobin agar. Heart blood: Growth of Gram-negative bacteria which
ferment all sugars tested.
In smears from liver , chains and groups of diplococci are observed, some of
them being situated in mononuclear leucocytes. In smears of spinal fluid and
heart blood no cocci and no protozoal organisms are observed.
Personally, I saw no cases of cerebro-spinal meningitis of an
ordinary type, but Scott described (1913), as already mentioned,
several such cases in connection with his investigations on 1 vomiting
sickness.* As a further proof that Diplococcus meningitidis occurred
in Jamaica at the same time that ‘ vomiting sickness ’ was prevalent,
I give abstracts of two observations, for which I am indebted to
Colonel Wilson, R.A.M.C.
Case I. This was a case in a Sergeant-Major who was taken ill on the 2nd
January, 1913, and died on the 6th. Clinical symptoms of meningitis were present
and there was a rise of temperature during the first days of the disease, with an
intermission on the 3rd and 4th day and a terminal rise up to 107*2 just before
death. The post-mortem examination showed an incipient meningitis, the
cerebro-spinal fluid being turbid 4 almost purulent.’ The spinal fluid contained
Gram-negative diplococci which did not grow on human Pleuritis serum.
Case II. This case occurred in the Chaplain of the Forces who was taken ill
on the 15th January, 1913. He presented marked clinical symptoms of meningitis,
and the disease lasted for about ten days. The patient recovered. Meningococci
were not demonstrated in the spinal fluid but developed in cultures from the
naso-pharynx.
Group E is composed of three cases, in which anatomically a
commencing meningitis was present, but no meningococci were found.
In Case 17 an adult woman died after an illness lasting for about
60 hours, during which she had presented symptoms fairly typical of
‘ vomiting sickness her five years* old child was taken ill the same
day as the mother, and died within six hours. The child did not
come under my observation, but I am indebted to Dr. Edwards for
the notes which are given below. The other patient in this group,
Case 35, was a girl of 11 years, living in the same house as Case 36,
435
a boy of II. Both children were taken ill at the same time, with
the same symptoms, and the duration of the illness was nearly the
same in both cases. All these circumstances evidently point to the
same disease having attacked the two children; in spite of all, the
two cases must be classified under different headings, as the results
of the post-mortem examinations, which were performed at the same
time, differed considerably. In Case 35 there was a marked, though
commencing inflammation of the meninges, but apparently no
pathological changes in the stomach or intestines. In Case 36, on
the other hand, there was a well marked haemorrhagic gastritis,
whilst the meninges showed only slight diffuse hyperaemia, a
common phenomenon in acute infectious diseases. Case 36 is
therefore discussed as belonging to the following group of cases
‘suspicious of yellow fever,* whilst Case 35 is considered, as it
anatomically must be considered, as one of meningitis. In the
latter case staphylococci were cultivated from the spinal fluid, but
it appears extremely unlikely that this occasional occurrence of a
very common organism should be of any etiological importance.
All circumstances considered, it is probably correct to regard the
two cases, 17 and 35, as meningitis of unknown origin, but
belonging essentially to the same disease as is represented by the
cases.
Thus, it may be that the separation of the two cases from those
in Group A occurring in the same places and under identical
circumstances is only provisional. Case 62 also somewhat resembles
the type in Group A.
Case 17. M.H., 27 years, St. Andrews. Admitted January 29, 3.10 p.m.
Public Hospital, Kingston, having vomited for the past 36 hours, complaining of
headache and pain in the limbs.
Whilst in hospital, no vomiting occurred, but patient had some mucopurulent
expectoration. Temperature 99 0 . Pulse about 80—90, regular and full to begin
with, apparently with somewhat increased tension, but on the morning of January
30 it was weak and irregular. She was conscious during the first day, but the last
morning she was in a state of quiet delirium and gradually passed into coma. There
were no shrieks, but some grinding of the teeth. No muscular rigidity, and no
Kernig’s symptom. Knee-jerk slightly increased on the left side, but could not
be produced on the right. No convulsions.
Spinal puncture made on January 30, about noon. Clear watery fluid comes
out, rapidly dripping; about two c.c. taken.
Patient died at 2.15 p.m., January 30.
Post-mortem examination : (With Dr. Scott) at 3.30 p.m., on January 30.
436
Intense hyperaemia of cerebral pia f less pronounced of spinal pia. Some
oedema and beginning fibrinous exudate over the cerebral hemisphere, not on the
basis, nor on the spinal cord.
The stomach shows slight oedema of the mucosa, and in some parts slight
capillary hyperaemia ; a pin head-sized superficial ulcer on the posterior surface,
near the lesser curvature.
Anatomical diagnoses: Leptomeningitis cerebralis incipiens. Metamor¬
phosis parenchymatosa et adiposa myocardii. Ulcus ventriculi parvum. Gastritis
acuta l.g. Nephritis acuta parenchymatosa. Perisplenitis chronica fibrosa.
Metamorphosis adiposa et parenchymatosa hepatis. Pancreatitis parenchymatosa
acuta. Helminthiasis intestinalis.
Microbiological examination :—
Nasgar tubes, spinal fluid: remain sterile.
This patient was the mother of a child which was taken ill and
died on January 28th, i.e., the same day that the mother’s illness
began. This case was observed by Dr. Edwards, to whom I am
indebted for the following particulars, which show that the case,
if classified, would have been included in Group A.
L.F., 4 years, 10 months, was taken ill at 4 p.m. Vomited several times
convulsions set in and coma and death supervened at 10 p.m. Dr. Edwards
performed the post-mortem examination and found:
Conjunctivae slightly jaundiced. Pleurae, lungs, heart, pericardium, peri¬
toneum, liver and spleen normal. Kidneys pale, the stroma show slight conjugation
of the cardiac portion, no blood in the lateral ventricles of the brain. Meninges
not congested.
Nasgar tubes were inoculated at the post-mortem examination with the
spinal fluid . They remained sterile.
Case 35. E.M., 11 years, black, Tollgate, with Dr. Earle. Living in same
house as No. 36. May Pen.
The child was healthy until Tuesday, February 4, 1913, at about 3 p.ra,
when she started vomiting some black substance, first declared so by grandmother,
who observed the child, but on further questioning she declared that the vomiting
had been colourless ; she continued vomiting until the following afternoon, when
the vomits ceased, and soon after convulsions, retraction of the head, and contraction
of the jaw were observed. She died at 4 p.m., on the 5th, without medical
attendance.
Post-mortem examination :—
Dura mater cerebralis normal. Pia mater shows slight diffuse hyperaemia
and oedema, and beginning fibrino-purulent exudation around some of the vessels
on the convex surface. Brain substance of normal appearance.
Anatomical diagnoses: Leptomeningitis cerebralis incipiens. Lympho-
adenitis mesenterialis simplex. Splenitis acuta. Nephritis acuta. Pancreatitis
parenchymatosa acuta. Helminthiasis intestinalis. Cadaverositas universalis.
Microbiological examination :—
Spinal fluid: Nasgar tubes, no growth of diplococci, but of Gram-positive
staphylococci.
Case 62. R.A.L., 9 years, black, seen, after death, with Dr. C. R. Edwards
at Kingston Public Hospital, March 9, 1913.
437
Post-mortem examination :—
Lumbar puncture gives a few c.c. of clear fluid, with admixture of blood.
Cerebral sinuses distended with blood clots ; pia considerably hyperaemic, especially
on convex surface and with slight sero-fibrinous exudation at the basis. Lateral
ventricles not distended, contain a very small quantity of serous fluid. Spinal
dura normal; pia very slightly hyperaemic in lumbar portion and slightly pig¬
mented. Brain and cord normal.
Urine : Contains traces of albumin and nucleo-albumin.
Anatomical diagnoses : Leptomeningitis cerebralis incipiens. Splenitis acuta.
Enteritis follicularis. Nephritis parenchymatosa acuta l.g. Petechiae epicardii.
Infarcti haemorrhagici pulmonis dextri. Sequelae pleuritidis duplicis. Adenitis
bronchialis caseosa. Endarteriitis aortae incipiens. Helminthiasis intestinalis.
Pediculosis capitis.
Histological examination:
Cerebral basis : The pia mater shows microcellular infiltration, no organisms
are observed.
Spleen : Considerable hyperaemia.
Liver : Moderate fatty change, oedema of stroma.
Pancreas : Both the alveolar epithelia and the islets of Langerhans of normal
appearances. The latter in particular are very well preserved.
Kidney: The epithelia of the convoluted tubules show granulation metamor¬
phosis, and slight necrobiosis, also slight vacuolization in the basal portions of
many of the cells. In osmic acid preparations minute fat droplets are seen, chiefly
in the basal portions of the epithelia of the convoluted tubules, but also in other
parts of these cells and in other cells, the epithelia of Bowman’s capsules and the
glomerulus-cells also being attacked.
Duodenum: Considerable microcellular infiltration of the mucosa. The
lymphoid follicles are considerably enlarged, numerous mast cells are present*
Lung infarct: alveoli filled with blood of apparently normal composition,
slight epithelial desquamation and microcellular infiltration of stroma in
neighbouring parts.
Microbiological examination :—
Nasgar tubes inoculated with spinal fluid and heart blood remain sterile.
Group F is very important. It contains cases in which one or
several symptoms gave rise to a suspicion of yellow fever. The
suspicious symptom which has been chiefly considered is black
vomit. Petechiae in the gastric mucosa may easily occur in other
conditions, and fatty liver is common in acute infections in children.
Therefore, these two phenomena have not been considered sufficient
to justify the inclusion of cases in this group, not even if supported
by the presence of acute nephritis. All these lesions are so common
in * vomiting sickness/ that they must really be considered in the
general discussion. On the other hand, dealing with extremely
acute cases, the possibility must be admitted that jaundice may not
have had time to develop. As it, moreover, is known that jaundice
is not invariably present even in fatal cases of yellow fever, the
438
presence of this symptom was not considered necessary in order to
include a case in this group. It is true, of course, that also black
vomit is no absolutely constant symptom, but it would not be likely
that it should be absent in rapidly developing and fatal cases.
The observation of black stomach contents at the post-mortem exam¬
ination is regarded as equivalent to the occurrence of black vomit.
In this way, and considering the general characters, the following
five cases were regarded as suspicious: Nos. 23, 36, 38, 47 and 54.
Notes of all these cases are given below few reference. In Case 23
there is little doubt that yellow fever is the correct diagnosis.
No complete clinical history could be obtained, as the patient, a
Chinaman, was unable to answer questions when I saw him, and his
Chinese friends explained themselves somewhat poorly in English.
Thus we could not find out whether the patient had suffered from
febrile symptoms, though it was evident that he had complained
of severe headache. It was also ascertained beyond doubt that
black vomit had been present, and the same could be concluded
from the aspect of the tongue. Examination of the blood showed
absence of malarial parasites, but a few intracorpuscular bodies
were found of the type of Paraplasma flavigenum. The post¬
mortem findings were typical of yellow fever in the essential details;
the small whitish patches which were found in the cerebral pia mater
were possibly remnants of some old meningeal affection, but there
was no fresh inflammation. The appearances of stomach, liver and
kidney were those met with in yellow fever. The liver showed,
microscopically, marked fatty and necrobiotic changes, in some
places of the Rocha-Lima type.* No meningococci developed in
the cultures, but diplococci were found in smears from various
organs, an observation which will be referred to later on.
The anatomical findings in Case 36 are very suspicious of
yellow fever, in spite of the absence of jaundice. Considering,
however, that gastric and pericardial petechiae were frequently
observed also in cases of ‘vomiting sickness’ which were not
suggestive of yellow fever, I should not feel confident in making
* R°«ha-Lima ( 1 9 1 z ) described a type of necrobiosis, combined with fatty change, in
k u r ’ w “ lc ". “f considers characteristic of yellow fever; the essential characteristic is
that the necrobiotic changes are mostly pronounced in the intermediate portions of the
lobules. This type is, according to my experience, common but not constant in yellow fever.
439
a diagnosis of yellow fever in such a case in the absence of a typical
clinical history and of an examination of blood taken during life.
Case 38 is similar to Case 36, with the additional symptom of
jaundice, thus strengthening the suspicion of yellow fever, but even
thus the evidence cannot be regarded as conclusive.
The Cases 47 and 54 are also essentially similar. The symptoms
are not suggestive of yellow fever, but rather of a meningeal
affection, though not markedly so. In Case 54, however, black
vomit was observed; in this case the peripheral blood was examined,
and no Paraplasnta flavigenum was found.
All these cases illustrate my previous statements about the
difficulties which beset the post-mortem diagnosis of yellow fever.
In all four cases the pathological lesions point to this diagnosis, but
the clinical and epidemiological characters make one hesitate in
pronouncing it, in the absence of conclusive parasitological
evidence.
Case 23. C.C., male, about 20 years, Chinese, Charlestown. Seen, on
January 31, 1913, with Dr. H. J. George.
The patient had lived in Jamaica from when he was a boy, and had been in
good health up till present illness.
He felt ill on January 27, started vomiting at about 10 a.m., to begin with
yellowish, later on greenish, and on January 30 and 31, red and black. He was
seen by Dr. George on January 31, in the morning; the temperature was then
normal. An injection of meningococcus-vaccine was given. Near midnight, on
the same date, I saw the patient, together with Dr. George. He was then in a
semi-comatose condition, not answering questions. Temperature was normal,
and it was not quite clear whether he had been febrile, but it was stated that
he had suffered from headache, besides the vomiting. Pulse about 90, weak, and
slightly irregular. Respiration stertorous. No enlargement of spleen or liver. No
abdominal tenderness. Perhaps slight jaundice of conjunctivae. The tongue
has red borders, but is otherwise covered with a blackish fur.
On February I, at 8.30 a.m., the patient arrived at the hospital in Buff Bay,
in an extremely weak condition. The temperature was not taken, but the skin
felt very cold to the touch. Pulse was filiform. There was no vomiting, but
enema provoked a greenish-black evacuation. The patient did not react on
stimulating treatment, and died at 11.30 a.m.
Post-mortem examination , at 2 p.m., hours after death.
Well nourished, fat body. Rigor present. Livores are very extensive on
back and sides, and are also present on the front of the femora. Slight jaundice.
: . Spinal puncture gives blood-stained fluid. Spinal cord not examined.
Brain . Dura intensely hyperaemic, pia likewise hyperaemic, but less pro-,
nouncedly. No liquid exudate, but on the convex surface some whitish patches,
in one place more yellowish. Brain preserved unopened, but right ventricle
punctured and nasgar tube inoculated with point of needle.
Abdominal cavity contains no fluid. Peritoneum of normal aspect. Abdom¬
inal organs normally situated.
+ 4 °
Spleen 13 x 11 X z\ cms. Capsule smooth, transparent. Colour, structure,
and consistence normal.
Kidneys 12 x x 3 cms. Capsule easily detached; Surface grey with a
few dilated stellate veins. Cortex grey, with some yellowish patches. Pyramidal
bases hyperaemic. Consistence normal.
Liver 30 x 15 X 7 cms. Surface yellowish grey, with hyperaemic patches.
Capsule smooth, transparent. Cut surface uniformly yellowish grey. Consistence
slightly diminished, pasty.
Pancreas of normal size and consistence, slightly hyperaemic.
The stomach contains a considerable amount of greenish fluid, and, adherent
to the mucosa, black, coffee-ground like, mucous matter, with numerous blood-
streaks. The mucosa shows considerable hyperaemia, chiefly of the posterior wall,
and numerous petechiae, especially on both sides of the lesser curvature, and on
the posterior wall. The duodenum contains greenish black fluid, and its mucosa
is slightly hyperaemic. In the middle portion of the jejunum there is a small
ecchymosis, and in the remaining portions of the bowel some hyperaemic patches
are observed.
The bladder contains about 100 c.c. yellow, slightly turbid urine. Mucosa
normal.
No liquid in pleurae ; but adhesions over the superior lobe of the left lung.
The lungs show intense hyperaemia of postero-inferior portions ; no oedema,
no infiltrations, no infarcts. No definite signs of tuberculosis in lungs, or bronchial
lymph-nodules.
Mucosa oj trachea hyperaemic.
Thyroid body of normal aspect.
Pericardium contains no excess of fluid. No petechiae.
Pulmonary artery free.
Endocard and myocard of normal aspect.
Urine : Slightly turbid, acid, contains albumin; no sugar ; faint reaction for
bile-pigments; numerous hyaline and granular casts; a few leucocytes and
erythrocytes ; numerous epithelial cells.
Anatomical Diagnoses: Gastritis ecchymotica. Metamorphosis adiposa
hepatis m.g. Pancreatitis parenchymatosa acuta. Nephritis parenchymatosa
acuta. Hypostasis pulmonum m.g. Leptomeningitis cerebralis veteris dati l.g.
Icterus universalis l.g.
Histological examination :—
Liver : Extremely intense vacuolization of cells (fat), as a rule most marked
around portal vessels. Marked necrobiotic changes which are often most intense
in the places of advanced fat changes around the portal vessels, but in other parts
most intense in the intermediate zone. Considerable capillary hyperaemia,
irregularly distributed. Some microcellular infiltration of the periportal
connective tissue.
PMcreas: Hyperaemia and slight haemorrhagic infiltration; oedema of
stroma. Marked necrobiotic changes, the cells in Langerhans’s islets being as
seriously affected as those in the parenchyma proper.
Kidney ; Vacuolization of basal portions of cells (fat). Some increase of
stroma which is oedematous ; slight microcellular infiltration, considerable
hyperaemia. Coagulated serum in Bowman’s capsules around the retracted
glomeruli.
441
Microbiological examination :—
Nasgar tubes, spinal fluid: Sterile. Nasgar tubes, ventricular fluid, Gram¬
negative bacteria, no diplococci. In smears of peripheral blood a few intracellular
ring-shaped organisms are found of the type of Paraplasma flavigenum.
No protozoal organisms are found in smears of organs, but in smears from
spleen and liver various bacteria are observed; amongst these are diplococci which
are Gram-negative and in several instances intracellular.
Case 36. P.B., 11 years, black, Tollgate, with Dr. Earle, May Pen. Living
in the same house as No. 35.
The boy had been in poor health previously and often had ‘ fever.* He was
taken ill on Tuesday, February 4, 1913, at about 3 p.m., when he began to vomit ;
he vomited all night and during the next day, until he died near midnight on the
5th, without medical attendance. He had no convulsions, but retraction of the
head was observed by the mother. During his previous illness he had never suffered
from vomiting.
Post-mortem examination: At 8 p.m., on February 6, 20 hours after death.
Poorly nourished body. Rigor present. No jaundice. No cutaneous affections
or parasites.
No spinal fluid obtained by lumbar puncture; spinal cord not examined.
Dura mater cerebralis normal. Pia mater shows slight acute hyperaemia over all
parts of the brain. No oedema or exudation. No distension of lateral ventricles ;
a small quantity of clear fluid is seen at the basis of the skull. Brain substance
shows normal aspect.
The abdominal cavity contains no fluid. The peritoneum is of normal aspect.
The abdominal organs are normally situated.
The mesenteric lymph nodules are considerably enlarged and hyperaemic, of
normal consistence.
The gastric and intestinal vessels are considerably distended. There are a
few easily detached intussusceptions of the small bowel. Tongue coated with
white fur. No haemorrhage of gums. Tonsils slightly enlarged ; no signs of acute
inflammation. Oesophagus normal. The stomach contains brownish, apparently
haemorrhagic fluid ; the mucosa is hyperaemic and shows on a small portion of the
posterior surface, close to the lesser curvature numerous petechiae. The duodenum
is hyperaemic, otherwise normal; jejunum and ileum, show hyperaemic patches ;
no haemorrhages, and no swelling of Peyer’s patches, or solitary follicles; no
ulcers. No intestinal parasites.
Liver : 22x13x6 cms. Capsule smooth, transparent; colour pale grey
with yellowish patches ; consistence distinctly diminished. The duodenal papilla
presents a small, prominent mucous mass which, however, gives way when pressure
is exerted on the gall bladder, and the bile then flows out freely; mucosa of bile
ducts and gall bladder normal; bile remarkably dark, and somewhat viscid.
Pancreas is hyperaemic, and of considerably diminished consistence.
Several lymph nodules at the porta hepatis are enlarged and show haemorrhagic
infiltration.
Spleen: 19 x 5 X ij cms. Capsule smooth, slightly thickened; tissue
somewhat pale, with slightly prominent follicles.
Suprarenal capsules normal.
Left kidney 8x5x2 cms. Capsule easily detached; surface is smooth,
shows capillary hyperaemia, but no distension of stellate veins; cortex pale
yellowish ; pyramidal bases considerably hyperaemic. The right kidney presents
the same characters as the left. Mucosa of renal pelves, ureters, and bladder
normal.
442
The pleura* contain no fluid and show no adhesions. The lungs are pale,
showing only a very slight hyperaemia of the posterior portions; the lung tissue
contains air throughout; there are neither oedema, infiltrations, nor infarcts.
The pericardium contains a few drops of clear fluid. The visceral pericardium
shows a considerable number of minute petechiae and small ecchymoses on the
anterior, lateral, and posterior surface of the left ventricle, especially close to the
auricle, and also a few similar ones on the auricle. Heart conus 7 x 7 X 2 J cms.
Endocard and valves normal. Myocard pale grey, with more yellowish patches;
consistence distinctly diminished.
The larynx and trachea are normal; the smaller bronchi contain mucous
fluid.
The thyroid body and the hypophysis cerebri are of normal size and aspect.
Anatomical diagnoses: Gastritis ecchymotica. Hyperaemia intestinorum.
Metamorphosis adiposa hepatis. Pancreatitis parenchymatosa acuta. Nephritis
parenchymatosa acuta. Lympho-adenitis mesenterialis et portae hepatis. Peri¬
splenitis chronica l.g. Ecchymoses pericardii. Metamorphosis adiposa myocardii.
Hypostasis pulmonum l.g.
Histological examination :—
Spleen : Hyperaemia ; oedema of stroma.
Pancreas : Localized necrobiosis ; oedema of stroma.
Kidney : Localized hyperaemia, granular change and karyolysis in many cells
in the convoluted tubules. Some basal vacuolization.
Stomach : Some increase of lymphoid tissue and moderate diffuse microcellular
infiltration ; oedema of submucosa.
Duodenum: Hyperplasia of Brunner’s glands and lymphoid tissue, slight
microcellular infiltration. Necrobiotic changes in the mucosa.
Brain : Hyperaemia and slight superficial microcellular infiltration.
Case 38. D.L., 8 years, black, admitted to Kingston Hospital in a dying
condition, on February 7.
The boy had been ill with fever and vomiting. No further particulars were
obtained.
Post-mortem examination , on February 7, at 12.30 p.m.
Anatomical diagnoses: Icterus universalis l.g. Gastritis ecchymotica c.
erosionibus. Metamorphosis adiposa hepatis. Nephritis parenchymatosa acuta
c. metamorphosi adiposa. Metamorphosis adiposa myocardii. Hyperaemia piae
matris cerebralis. Pancreatitis parenchymatosa acuta.
Black and blood-stained mucus is adherent to the wall of the stomach, which
contains a few milk-clots.
A few slightly hyperaemic patches are seen in the small and large bowel. The
Peyer’s patches are likewise slightly hyperaemic.
Liver: 650 grms., yellowish grey, homogeneous, pasty.
Kidneys of normal size ; capsule easily detached; surface congested, with
yellow patches ; cortex yellowish ; consistence diminished.
Spleen of normal aspect. Slight diffuse hyperaemia of pia cerebralis, no
odema or inflammatory exudate.
Urine : The bladder contains a considerable quantity of pale urine, which
contains a small quantity of albumen, no sugar, no bile-pigments, few hyaline
casts, numerous epithelial cells, a few erythrocytes and leucocytes.
Histological examination :
Lymph nodules : Hyperaemia and perinodular lymphoid infiltration.
Myocard : Numerous small vacuoles (fat) in muscle fibres.
443
Liver: Advanced fatty metamorphoses, all zones being equally affected.
Marked anaemia.
Pancreas : Advanced necrobiotic changes, most nuclei being invisible and
cell limits poorly defined.
Kidney Large vacuoles (fat) occupying basal parts of cells in convoluted
tubules. Most nuclei staining well, straight tubules not affected. Marked
anaemia.
Medulla : Oedema and slight infiltration of pia.
Case 47. A.W., 20 years, black, Windward Road, Doncaster Pen. Seen in
Public Hospital, Kingston, with Dr. Thomson.
Admitted in a weak condition. History of sudden onset of vomiting last
night. No history of previous illness. Her infant, if years old, mentioned in
note to case 48, was attacked earlier in the evening, and died. A nephew also
started vomiting during the night (case 48).
Temperature 98*4° on admission. Pulse small and irregular. Patient
complains of pain in the epigastrium.
Temperature went up in the afternoon to 99 0 and ioo°, but in the evening
again down to 98°, where it remained until the time of death. Heart’s action
was in the afternoon regular; sounds clear. Lungs clear. Tenderness in epigas¬
trium. Spleen not felt. Pupils equal, not contracted, nor dilated, react to
light and accommodation. No vomiting since admission, but drowsiness. No
rigidity; Kernig’s symptom not present.
February 19. During the afternoon patient’s condition grew weaker; the
drowsiness passed into coma. The breathing became laboured; the pulse was
very small and irregular, but responded readily to stimulation. No rigidity of
limbs.
At 4 a.m., patient appeared to be sinking ; she was quite comatose, with
Cheyne-Stokes’s respiration ; pulse small and irregular.
Lumbar puncture gave clear, watery fluid. Urine could not be collected.
Death at 5.35 a.m.
Post-mortem examination at 7 p.m., on February 19, 13 hours after death.
Body well nourished; rigor present; no jaundice. A few Pediculi capitis ,
no other parasites, nor affections of the skin.
Axillary lymph nodules moderately enlarged and hyperaemic ; inguinal enlarged,
cervical normal.
The subcutaneous fat shows, in various places, small haemorrhages.
Dura spinalis normal; pia hyperaemic, especially in lower part. Spinal cord
normal. Cerebral dura normal; pia considerably hyperaemic, particularly on
the convex surface. No exudation, but some haziness over the basis and over
the Sylvian fissures. Lateral ventricles not distended, contain a few drops of clear
fluid. Brain substance normal.
Hypophysis slightly enlarged, hyperaemic, and with soft, yellowish patches.
Pleurae normal. Lungs slightly hyperaemic in postero-inferior parts, contain
a few minute haemorrhagic infarcts. Smaller and larger bronchi, trachea and
larynx show diffuse hyperaemia and some mucous secretion. Bronchial lymph
nodules not enlarged.
Pericardium contains a few c.c. of serous fluid; conus 9x11 X 5 cms.;
epicard shows several small ecchymoses on posterior and left surface of left ventricle
and auricle. Endocard and valves normal. Myocard shows small yellowish
patches; consistence somewhat diminished; left ventricle fairly well contracted*
Thyroid body normal*
444
Peritoneum normal, without fluid. Abdominal organs normally situated.
Spleen: Small, n X 5 X 2 cms. Capsule normal; colour pale; follies
grey, slightly prominent.
Suprarenal capsules normal.
Mesenteric lymph nodules slightly enlarged, not hyperaemic.
Left kidney : 12 x 6 x 4. Capsule easily detached ; surface extremely pale
except in a few places where there is some dilatation of the stellate veins. Cortex
pale, yellowish grey, and some of the septa completely yellow, of almost caseous
aspect, the parenchyma structureless, bulging on the cut surface. Pyramids pale.
Consistence of the whole organ considerably diminished, and of the yellow parts,
soft, pasty. Right kidney 11 X 7 X 3 cms.; characters similar to those of he
left. Mucosae of renal pelves, ureters and bladder normal. The bladder contans
a small quantity of pale urine.
Liver: 29 x 15 X 8 cms. Capsule normal; surface pale, yellowish; tissie
yellowish grey, homogeneous; consistence pasty.
Pancreas of normal size, but of somewhat diminished consistence.
Tongue covered with white fur. Tonsils normal.
The stomach contains a large quantity of reddish black fluid, with bltck
particles, coffee-grounds like, also some blackish mucus (the fluid is acid, and gves
reactions for haemoglobin); mucosa hyperaemic; no haemorrhages visiole.
Duodenum contains considerable quantity of dark, sanguinolent fluid, mucosa
intensely hyperaemic, with minute petechiae. Jejunum normal. Ileum hyper¬
aemic. Colon shows marked hyperaemia. Appendix 18 cms. Two ascarides
in ileum.
Anatomical diagnoses: Hyperaemia meningum. Hypostasis et infarcti
haemorrhagici parvi pulmonum. Ecchymoses pericardii. Metamorphosis
adiposa myocardii. Ecchymoses telae adiposae subcutaneae. Gastritis acuta
haemorrhagica. Duodenitis acuta haemorrhagica. Hyperaemia coli. Meta¬
morphosis adiposa hepatis maximo gradu. Nephritis acuta duplex parenchymatosa
c. metamorphosi adiposa m.g. Metamorphosis adiposa (?) pancreatis. Lymph¬
adenitis axillaris, inguinalis et mesenterialis acuta. Pediculi capitis. Helmin¬
thiasis intestinalis.
Histological examination :—
Axillary lymph nodules : Hyperaemia.
Pituitary body; Hyperaemia.
Suprarenal capsule : Diffuse hyperaemia, especially intense in the periglandular
tissues where also infiltrating haemorrhages are observed.
Liver: Intense fatty metamorphosis. Necrobiotic changes are present
throughout the organ; in some places these phenomena show the Rocha-Lima
type, but most often only insignificant remains of protoplasm are preserved in the
cells surrounding the hepatic veins, where the affection in many places appears
mostly advanced. There is slight microcellular infiltration of the periportal
tissues.
Pancreas: Minute fat droplets are observed in the alveolar epithelia and
slightly larger droplets in the cells of Langerhans’s islets.
Kidney : The epithelia of the convoluted tubules show granulation of pro¬
toplasm and their basal portions are occupied by vacuoles which correspond to fat
droplets in osmic acid specimens. These phenomena are present with some
irregularity in nearly all the convoluted tubules and exceedingly intense in many
of those of the first order. They are also very intense in the ascending limbs of
Henle’s loops, but there is hardly any fatty change of the cells in the descending
limbs and collecting tubules. A number of patches of intense capillary hyperaemia
H5
arc observed and in the neighbourhood of these patches there is some microcellular
infiltration. There is moderate increase of the stroma especially of the pyramids,
the glomeruli are slightly retracted and minute fat droplets are observed in their
cells.
Myocard : Moderate fatty change.
Microbiological examination :—
Spinal fluid, nasgar tubes : Gram-negative diplococci which change glucose,
galactose and maltose, and saccharose slightly, but not lactose and mannite ; also
some colonies of Gram-negative cocco-bacilli.
Heart blood, nasgar tubes : Gram-negative cocco-bacilli. Smears of peripheral
blood and heart blood show no protozoa but in the heart blood some large bacteria.
No organisms are seen in smears from spinal fluid after centrifugalization.
Case 54. D.A., 7 years, coloured, Eastward Villa, Hagley Park. Seen, after
death, with Dr. C. R. Edwards, Halfway Tree, on February 25 and 26, 1913.
The child has never been ill before. She felt slightly unwell on February 23,
in the evening, was crying and complaining of pain in the stomach; during the
following night she woke up, at 1.30 a.m., February 24, groaning, and had three
attacks of convulsions with clonic spasms. During the next day she vomited
several times clear fluid, and did not take any food; she did not seem to have
fever, but, according to the mother, the skin turned yellow (vide infra). She was
awake during the whole of the following night, restless, semi-unconscious, and
complaining of abdominal pain. At 9 a.m., on February 25, she was brought to
Dr. Edwards’ office, where she remained until her death. Whilst here, several (8)
convulsions were observed; they consisted of clonic spasms, with final tonic con¬
tractions ; she vomited repeatedly, to begin with clear, watery fluid, but later on
dark, coffee-grounds like matter, and reddish, thin fluid, with suspended small
particles, in large quantities. She was unconscious, became gradually weaker,
and died at about 4 p.m., just a quarter of an hour before my arrival.
When seen after death, the body appeared rather yellow, but there was no
trace of yellow in the conjunctivae. Spinal fluid, blood, urine and vomited
matter collected by Dr. Edwards.
Spinal fluid: About 20 c.c., clear.
Bladder contained only a few drops of pale urine.
Post-mortem examination, at 11 a.m., on February 26, 19 hours after death :—
Body somewhat poorly nourished. Rigor present. Extensive livid patches
on the back, invading even the flanks. No jaundice of conjunctivae, but the body
has a peculiar yellow colour. No skin affection, and no external parasites, except
lice in the hair.
Spinal venous plexus gorged with blood. Dura spinalis normal, cerebralis
slightly hyperaemic ; pia spinalis and cerebralis intensely hyperaemic, and the
latter slightly oedematous at the basis. Lateral ventricles not distended, contain
no fluid. Brain and cord normal.
Hypophysis softened, otherwise normal.
Inguinal, axillary and cervical lymph nodules considerably enlarged and intensely
hyperaemic.
Left kidney: 8x5x2 cms. Capsule easily detached; surface smooth,
dark, with capillary hyperaemia, shows a few yellow patches; cortex reddish
grey ; glomeruli normal; columnae Bertini pale, yellowish; pyramidal bases
hyperaemic; consistence considerably diminished. Right kidney shows same
characters as left.
Tonsils : Hyperaemic, contain several small pus foci.
446
Mucosa of pharynx swollen and hyperaemic.
Mesenteric lymph nodules considerably enlarged, not hyperaemic.
The stomach contains a considerable quantity of greenish black fluid, with
minute, brownish black particles ; blackish mucus is adherent to the mucosa, which
shows intense hyperaemia of cardiac portion of anterior wall, and numerous
petechiae. On the posterior wall are several small erosions, without any sign of
haemorrhage.
Duodenum: slightly hyperaemic. Jejunum shows patches of hyperaemia;
ileum pale, colon shows several hyperaemic patches.
Liver: 21 x 14 X 5 cms. Capsule normal; colour reddish grey, with yellow
patches; substance yellowish grey, with patches of pronouncedly yellow colour,
and a few patches of hyperaemia; consistence considerably diminished, and
somewhat friable.
Pancreas : Slightly enlarged; the caudal portion shows intense hyperaemia,
and some haemorrhage into the tissue; caput and corpus pale ; the whole organ
is of considerably diminished consistence.
Anatomical diagnoses : Hyperaemia meningum. Poly-lymphadenitis acuta.
Metamorphosis adiposa myocardii, et hepatis. Pancreatitis acuta. Nephritis
acuta incipiens duplex. Hypostasis pulmonum. Gastritis acuta ecchymotica.
Hyperaemia jejuni and coli. Elytritis. Endometritis. Cystitis ? Helmin¬
thiasis intestinalis. Tonsillitis acuta. Pharyngitis acuta.
Histological examination :—
Axillary lymph nodules: Slight diffuse hyperaemia, in some places more
pronounced. One considerable haemorrhage and several smaller ones.
Myocard: Fragmentation, numerous small fat droplets in fibres.
Liver : Diffuse hyperaemia, one or a few large vacuoles (fat) in nearly every
cell. Slight microcelluiar infiltration of periportal tissue and penetrating into
the lobules.
Pancreas : Advanced necrobiotic changes.
Kidney: Numerous fairly small fat droplets in the epithelial cells near the
basal membrane, mostly in the convoluted tubules, less in the straight tubules
and only very few cells in the glomeruli.
The spinal fluid is clear, alkaline, gives fairly intense albumin reaction.
The centrifugalized deposit consists chiefly of erythrocytes ; also some lymphocytes
and a few polymorphonuclear leucocytes are seen.
Microbiological examination :—
Peripheral blood: No parasites. Heart blood , nasgar tubes, long Gram-negative
bacteria. Spinal fluid, nasgar tubes, short thick Gram-negative bacteria which
change all the sugars tested. Vomits: The first portion examined is clear bile
acid; second portion is reddish black, with suspended dark brown particles. It
is intensely acid and contains erythrocytes (microscopical) and haemoglobin (Guajac
test).
Group G contains four cases: Nos. 6, 26, 45 and 58. Case 6 was
a typical case of infantile gastro-intestinal atrophia, and, when I saw
the case, there was no suspicion whatever of * vomiting sickness/
My attention was, however, drawn to this case, because it was said
to have been diagnosed as * vomiting sickness * about three weeks
previously. I regard this as an example of mistaken diagnosis,
which is apt to occur, especially in recovering cases. This case
447
I exclude absolutely from the discussion as far as 4 vomiting
sickness * is concerned, but it may be mentioned that even in this
case meningeal hyperaemia was present, contrasting with the marked
anaemia of all other organs. Case 26 was also one of general
atrophia, but this time in an old man. In this case Gram¬
negative diplococci were cultivated 1 from the spinal fluid, a result
which there is no reason to consider otherwise than as an accidental
post-mortem infection. Case 45 resembled the type of 4 vomiting
sickness/ but the presence of numerous malarial parasites affords
sufficient explanation of the symptoms and of the anatomical
lesions. This case serves to illustrate the difficulties of diagnosis
and is therefore given in full. Malarial parasites were also found
in Case 58, but in small numbers; the diagnosis of malaria as
causa eificiens must therefore be regarded with some doubt, and a
diagnosis of 4 vomiting sickness * has to be considered. The
patient in Case 58 was a sister to the one is Case 57 belonging to
Group B, as organisms of the meningococcus type were found.
The two sisters were taken ill and died at the same time, obviously
from the same disease; it seems, therefore, that the essentially
different results of the microbiological examinations must be
disregarded and both cases classified as ‘vomiting sickness/
Case 6 . J.R., 26 months, Porus. Ill for three weeks, with fever, vomiting
and diarrhoea with green stools; no convulsions, no shrieks.
Post-mortem diagnoses: Atrophia ventriculi et intestinorum. Anaemia
universalis. Hyperaemia piae matris cerebralis. Hypostasis pulmonum.
Case 26. R.L.D., about 60 years, black, from Belvedere, Portland, not seen
before death, but reported as having been taken ill acutely with vomiting. Later
it was found out that he had been weak for several months and had vomited for
some days before death. The corpse was brought down to the hospital mortuary
and a post-mortem examination performed, with Dr. George.
The autopsy on February 1, 1913, showed simple atrophy of all organs and
multiple cysts in both kidneys. There were no signs of cerebral meningitis and no
affection of the stomach. The spinal cord was not examined, but lumbar puncture
gave no fluid, except a single drop, with which a nasgar tube was inoculated.
Histological examination :—
Liver : Atrophia pigmentosa.
Microbiological examination :—
Nasgar tubes, spinal fluid : Gram-negative diplococci in groups which change
glucose, galactose, saccharose and maltose, but not lactose.
Case 45. M.K., 8 years, black, Spanish Town Road. Kingston Public
Hospital, February 15, 1913.
448
Admitted at 5 p.m., suffering from vomiting and convulsions, which had
started the same day, suddenly. Head retracted. Death at 7.20 p.m.
Post-mortem examination , at 12 noon, on February 16, about 17 hours after
death :—Body fairly well nourished. No skin affection. Hyperaemia of lips
and gums; small ulcers on lips. No jaundice. Rigor present.
Cervical and axillary lymph nodules enlarged and hyperaemic, inguinal
enlarged.
Lumbar puncture gives only one or two drops of serous, slightly blood-stained
fluid. Spinal dura normal, pia hyperaemic; cord normal in dorsal portion,
the only part examined. Cerebral dura slightly hyperaemic, pia considerably
hyperaemic, especially on convex surface; a few drops of serous fluid at basis;
no fibrinous exudation, one or two c.c. of clear fluid in lateral ventricles; brain
substance normal, slightly pale.
Hypophysis small, 6x4x3 mms., normal in appearance.
Pleurae normal. Lungs show slight hyperaemia, most marked in postero-
inferior parts. Bronchi hyperaemic, contain some mucous secretion; trachea
and larynx normal. Bronchial lymph nodules slightly enlarged and anthracotic.
Pericardium contains no fluid; epicardium, endocardium and valves normal.
Heart conus 6 x 7 X cms.; left ventricle fairly well contracted. Myocardium
normal. Pulmonary artery and aorta normal.
Thymus 6x4x1! cms., normal, in appearance; consistence slightly
increased.
Thyroid body normal.
Peritoneum normal without fluid. Abdominal organs normally situated.
Spleen : 15x9x5 cms. Capsule slightly folded and thickened ; substance
homogeneous dark purplish grey, without prominent follicles. Consistence
considerably diminished.
Mesenteric lymph nodules slightly enlarged and hyperaemic.
Suprarenal capsules : Considerably hyperaemic, the left most pronouncedly
so; the left is also slightly enlarged and contains several small haemorrhages
Right kidney : 8 x 4! X 2} cms. Capsule easily detached ; surface smooth,
with capillary hyperaemia; some yellowish grey patches on surface; cortex
hyperaemic in superficial part, grey in columnar portion. Slight hyperaemia of
pyramidal bases. Left kidney shows more marked hyperaemia.
Renal pelves, ureters and bladder slightly hyperaemic ; bladder contains
about 30 c.c. of clear yellow urine.
Genital organs normal.
Liver: 25 x 16 X 5 cms. Capsule normal; surface dark purple, with a
few small yellowish patches; structure and consistence normal. Bile ducts
patent and normal; gall bladder contains a considerable quantity of yellowish
bile.
Pancreas normal in size; slight hyperaemia; considerably diminished
consistence.
The tongue's entire surface is covered with a whitish fur. Left tonsil slightly
enlarged, haemorrhagic infiltrated and contain some inspissated pus. Oesophagus
normal. The stomach contains alimentary fluid and mucus. The mucosa shows
a few hyperaemic patches, is otherwise normal. Duodenum shows hyperaemia
and swelling of follicles. Jejunum shows patches of hyperaemia and slight follicular
swelling in its upper part. The solitary follicles and Peyer’s patches in the lower
part of the ileujn are swollen, but not hyperaemic. Appendix normal. Colon
hyperaemic, with follicular swelling and dark, pigmented spots on follicles. No
worms
449
Anatomical diagnoses: Hyperaemia meningum, pulmonum, intestinorum et
capsulae suprarenalis. Splenitis acuta in chronica pigmentosa. Metamorphosis
adiposa l.g. Hepatis et pancreatis. Gastro-enteritis follicularis. Poly-lymph-
adenitis acuta.
Histological examination :—
Axillary lymph nodules: Moderate hyperaemia.
tonsils : Intense hyperaemia.
Liver : Slight fatty change.
Pancreas : Slight localized necrobiotic changes.
Kidney : Advanced necrobiotic changes of the epithelium of the convoluted
tubules with karyolysis and granulation of protoplasm. No vacuoles, and glomeruli
normal. Epithelia of straight tubules well preserved. Moderate irregularly
distributed hyperaemia.
Microbiological examination :—
Spinal fluid: Nasgar tubes remain sterile.
Heart blood , smears: Plasmodium vivax; abundant infection with young
parasites.
Spleen : smears ; Plasmodium vivax : abundant pigmentation.
Heart blood: Nasgar tubes remain sterile. I haemoglobin-agar growth of
Gram-negative cocco-bacilli.
Case 58. D.M.M., 4 years, black, seen, after death, with Dr. Peck, Spanish
Town, on March 5, 1913.
Post-mortem examination :—
Lumbar puncture gives clear fluid, with admixture of blood. Dura normal.
Pia slightly hyperaemic on lumbar portion of cord and convex surface of brain ;
normal at basis. Lateral ventricles normal. Brain and cord normal.
Cervical , axillary and inguinal lymph nodules , enlarged and slightly hyperaemic,
especially the axillary.
The stomach contains alimentary fluid; slight hyperaemia of mucosa.
Duodenum shows considerable follicular swelling. Jejunum normal, except
for a few hyperaemic patches. Ileum normal. Appendix and colon normal.
One Ascaris lumbricoides .
Anatomical diagnoses: Hyperaemia piae matris l.g. Lymphadenitis acuta
l.g. Hyperaemia pulmonum. Petechiae epicardii l.g. Hyperaemia ventriculi
l.g. Duodenitis follicularis. Nephritis acuta l.g. Helminthiasis intestinalis.
Urine : Acid; contains no albumin, sugar, bile pigment, or casts.
Histological examination :—
Axillary glands : Considerable hyperaemia, increase of stroma.
Spleen : Pigmentation.
Liver : Irregular hyperaemia ; marked fatty change.
Pancreas : Moderate fatty change.
Kidney : Moderate fatty change, some increase of stroma and some micro-
cellular infiltration. Epithelia well preserved.
Duodenum: Brunner’s glands prominent. Microcellular infiltration and
irregular hyperaemia.
Microbiological examination :—
Spinal fluid , nasgar tubes : remain sterile. Heart blood, nasgar tubes remain
sterile. Heart blood smears : Plasmodium vivax , sparse infection.
Group H includes five cases, Nos. 3, 21, 22, 28 and 53, in which
I was unable to obtain sufficient data to attempt a diagnosis. These
cases may be left entirely out of the discussion.
450
V. DESCRIPTION OF VOMITING SICKNESS
It is quite obvious, from the preceding report of cases, that not all
of those described are of the same nature. An attempt to determine
the nature of 4 vomiting sickness * may be preceded by a re-grouping
of the cases in order to establish a well-defined field for
discussion. So far, it is necessary to include amongst the
chief characteristics the negative one, that a diagnosis of
vomiting sickness is inadmissible as long as a case can be
diagnosed otherwise on clinical and pathological grounds.
The microbiological examination does not necessarily decide
this matter. Two or more diseases may be caused by the same
pathogenic agent and nevertheless differ so essentially from each
other that they are in all systems classified under different headings.
Table B
Yellow
Infantile or
Vomiting Sickness
Possibly Vomiting Sickness j
1 Fever
Malaria
Senile
Atrophy
Group
A
Group
B
Group
C
Group
D
1 r
Group
E
Of I
Group
F
Of
Group
G
°f
Group ;
F
Of !
Group
G
Of
Group
G
16
ii |
D
5
■
1
!
1 (or 5)
1
1 1
2
40 cases
l
13 cases
■ (or 5 )
cases
l
1 case
2 cases
Furuncles, osteomyelitis, and generalised sepsis are often due to
one and the same organism, but it would obviously be wrong to
consider them as identical diseases. Similarly, ‘ vomiting sickness'
might be caused by the same germ that is responsible for meningitis,
for instance, and still with some right claim a name as independent
disease. From this point of view we shall have to consider only
the cases in Groups A, B and C. Those in the other groups either
do not correspond to the type represented in A, B and C, or they
are insufficiently observed, or they can be otherwise diagnosed.
It appears that an arrangement like that given in Table B would
represent a natural classification of the provisionally established
groups, leaving out of consideration the non-defined group H.
45 1
The following 1 clinical and pathological description applies to
the cases which I would regard as 1 vomiting sickness/
Definition .—* Vomiting sickness ' is an acute infectious disease
which chiefly attacks children, often with fatal issue in a very
short time, and which has so far only been observed in blacks,
coloured persons and Hindoos in Jamaica.
Etiology .—The question of a specific pathogenic germ has not
yet been fully solved; it is discussed in the following section.
Deficient hyperaemic conditions, malnutrition and exposure to cold
have been mentioned by various observers as etiological factors of
importance, but there is no evidence that any of these factors should
be very essential. Personally, I feel inclined to eliminate mal¬
nutrition entirely from this list, as in my experience a considerable
number of cases have occurred in well nourished, even fat, and
absolutely healthy-looking individuals. The mode of transmission
is unknown, and such suggestions as have been made in this respect
are based more upon analogy than upon actual observation.
Distribution and epidemiology .—‘ Vomiting sickness ’ is, as far
as the majority of the cases are concerned, a disease of people
inhabiting poor huts more or less closely surrounded by bush.
This, however, is by no means a rule without exception; several
cases occurred in well-built and cleanly kept houses in villages, or
situated on dry ground, apart from the bush.
Cases were observed, in this as in earlier outbreaks, in con¬
siderable numbers in some districts, and were altogether absent in
others. This circumstance is, however, not one of very great
importance. Districts are large in Jamaica, and it is no easy matter
to trace the connection between ten and twelve cases scattered over
an area of nearly a hundred square miles. As a matter of fact,
a connection could rarely be demonstrated with certainty. When
several cases occurred in one and the same house, a connection might
of course be presumed to exist between them, but in each instance
one would sooner think of a common source of infection than of
transmission of the infection from one case to another, because the
cases as a rule occurred either simultaneously or in very rapid
succession. My material includes various examples of almost
simultaneous occurrence of vomiting sickness in two or more
members of one and the same family, living in the same house, as
452
in the following group of cases: Nos. 14, 15 and 16; 17 and child
not observed; 35 and 36; 47, 48, and infant; 57 and 58. On the
other hand, there are even more numerous instances of the isolated
occurrence of the disease in' one member of a family, although
several other children were living in the same house.
Symptomatology .—The duration of the incubation period is
unknown, but there is some reason to believe that it is short,
considering the hyperacute character of the disease. Nothing
definite has been recorded about the occurrence of prodromal
symptoms.
The illness usually sets in suddenly, often in the night, or early
morning, though this is by no means a constant feature; in fact,
it is not uncommon that the symptoms are first observed in the
evening and that death takes place during the night. Vomiting is
very often the first symptom; it is usually watery or consisting of
alimentary fluid, at any rate to begin with, but may later on become
bilious or perhaps even haemorrhagic. Vomiting is not a constant,
but an extremely common symptom; out of the forty cases in the
Groups A, B and C, its occurrence is mentioned twenty-four times,
and its absence six times; in the remaining ten cases no definite
information was obtained. The next symptom, as regards
frequency, is that of convulsions, which are sometimes described as
clonic, but in other cases as tonic contractions, lasting for several
minutes; an exact description of this symptom is at present hardly
possible, as most cases were not medically observed. For the same
reason, it is extremely difficult to discuss the occurrence and
frequency of other nervous symptoms, but it can confidently be
stated that coma is exceedingly common and that a sudden collapse
apparently occurs in some cases. Rigidity of the neck muscles was
not frequently observed, nor was rigidity of other muscles.
Kernig’s symptom was only recorded twice in the cases of the first
three groups, but both with regard to this and the two foregoing
symptoms it is important to remember that very few patients came
under medical observation at all or sufficiently early to make
a thorough examination possible.
Of other symptoms than those of a nervous character little is to
be said. The course of the illness is so rapid that most organic
lesions do not become recognisable clinically. Febrile temperatures
453
are seldom observed, but it seems probable that subnormal
temperatures occur, though there is little evidence of this. The
vomiting is, perhaps, in the majority of cases of cerebral origin,
but just as often one would believe it to be due to gastric irritation.
The severe renal affection which was frequently observed at the
post-mortem examination could hardly be expected to produce any
clinical symptoms; it may be specially mentioned that abundant
urine was often found at the autopsy and that the amount of
albumin was usually very small. It would appear that the
urine found had been excreted before the development of the
disease, or, at any rate, of the kidney affection. The rapidity with
which the illness reaches a fatal termination shows, approximately,
an inverse ratio to the age of the patients; infants die sometimes
less than an hour after the appearance of the first symptoms, while
cases in adults may last for one or two days.
Pathological anatomy .—The most striking anatomical lesions
are: enlargement and hyperaemia of the lymphatic nodules,
petechiae on the surface of the heart and in the gastric mucosa,
necrobiosis of the pancreas, liver and kidneys, especially of the
pancreas, and fatty change of the same and other parenchymatous
organs. There is often a marked hyperaemia of the spinal and
cerebral pia, but in other cases it was very moderate, and in others
limited to the cerebral pia alone. The intensity of the lesions
differed very much, especially that of the fatty changes, which was
most marked in the cases of one or two days’ duration in adults,
whilst it was seen in its initial stage in cases of extremely rapid
development.
General pathology .—The pathological physiology ought to
explain the most striking feature of the disease, namely, the rapid
course, the high mortality and the haemorrhagic degeneration and
necrobiotic phenomena just described.
All these phenomena are readily explained as the results of an
acute poisoning which may be due to the products of a bacterial
or protozoal organism. If the vomiting is considered of cerebral
origin, only the occurrence of cerebral symptoms has to be explained.
All these symptoms, vomiting, convulsions and coma, may be due
to an affection of the meninges, and this affection need not be a true
meningitis; they may just as well be the expression of a state of
‘ meningismus.’
454
Mortality and prognosis .—According to some observers (Tillman
and others, Annual Report ) a very large number of cases of
1 vomiting sickness * terminate in recovery. If this is true then
many other observers, including myself, have only seen, or at any
rate, have only diagnosed severe cases. This is possible, but for
reasons stated above I consider it more likely that a large number
of minor ailments, essentially different from ‘ vomiting sickness/
have been included in the statistics of the first-named observers.
However, this controversy cannot be decided at present, and it may
therefore be mentioned that the mortality has variously been found
as low as about 2 per cent., and as high as about 100 per cent.
In my own material, considering only the forty cases in Groups A,
B and C, with thirty deaths, the mortality is 75 per cent. If a
marked case of ‘ vomiting sickness ’ is seen, a very guarded
prognosis should be given; recovery may take place, but the
situation is always very serious.
Diagnosis .—Until the etiology of 4 vomiting sickness* is more
fully known, the diagnosis in vivo and post mortem must rest
exclusively upon the symptoms described above.
Treatment .—No specific treatment can be recommended with any
hope of success. Antimeningococcic serum might be used when
‘ meningococci * are found, but it is doubtful whether its use would
be justified in other cases. Moreover, as Scott (1913, 2) has
pointed out, its general use out in the country districts encounters
many difficulties of a practical nature. An anti-serum might, perhaps,
be prepared with the specific diplococcus, if this organism should
finally be proved to be the cause of vomiting sickness. Vaccine
treatment would hardly find any indication in this hyperacute
disease. Hot baths and stimulating injections (ether, camphor,
strychnine, etc.) may be recommended, but without much hope of
success, especially as there will usually be very little time for the
application of therapeutic measures.
Prophylaxis .—This question will be discussed in Section VII.
VI. THE NATURE OF VOMITING SICKNESS
At the onset of this discussion it is natural to ask, whether
diseases identical to ‘ vomiting sickness * occur outside of Jamaica.
Very little material is available which b^ars directly on this question.
455
In 1905 the Government of Jamaica made enquiries, in a circular
letter to the authorities of other West Indian Islands, with regard
to the possible occurrence of ‘ vomiting sickness * in other parts of
the West Indies. Only two of the answers are quoted by Ker in
the Annual Report (1906), the others being in the negative. The
one positive answer is from Finlay, who mentions an outbreak of
25 similar cases with 5 deaths, amongst soldiers, in Cuba: he
suggests that the disease is cerebro-spinal meningitis, no particulars
are given in the quotation. The other positive answer deals with
a disease (in Haiti) which has obviously no essential resemblance
to * vomiting sickness.*
Branch (1906), who has no personal experience with ‘ vomiting
sickness/ does not admit it as a morbid entity, and is of opinion
that similar cases occur elsewhere in the West Indies, carrying
different names, and representing in reality well-known diseases,
especially malaria and helminthiasis.
With regard to this opinion I must remark that it is difficult
without personal observation to realize what 1 vomiting sickness *
means. Before seeing cases, I quite naturally had the same idea
that it simply was something else; but the particular experience of
seeing one fatal case following another in children who a few hours
previously were playing out and who hardly developed any
symptoms before they died, appeared to me almost unique, and left
quite a different impression upon me.
Thus, until further evidence may be brought forward, we may
assume that ‘vomiting sickness* is a local disease of Jamaica.
Since the publication of Potter’s and Scott’s reports no further
papers have appeared on the subject. It will now be justifiable to
circumscribe the discussion, dealing, in the first place, with yellow
fever and meningitis only.
With regard to yellow fever, some circumstantial evidence is in
favour of this diagnosis. Jamaica was in earlier times one of the
principal foci of the yellow fever infection, a veritable hot-bed for
yellow fever. Even in this century, in 1904, a small outbreak
occurred in Port Royal, which was apparently of spontaneous
origin; this outbreak and the question whether there is any evidence
that isolated, autochthonous cases occur, is discussed by Potter
(1912).
456
Considering the question from an epidemiological point of view
there is obviously good reason for investigating very carefully any
disease which might prove an atypical form of yellow fever, and
which might thus not only establish the fact of yellow fever
endemicity in Jamaica, but also throw considerable light on yellow
fever epidemiology in general. On the other hand, epidemiological
evidence itself is almost sufficient to destroy this hypothesis.
Yellow fever in native children in endemic areas is probably always
a fairly benign disease; at any rate, there is no reason to suspect
that it should ever acquire such a malignant, fulminant type as that
of * vomiting sickness.’ The occurrence of these cases amongst the
native population only, without ever attacking foreigners, is also
very much against the theory of yellow fever. It might be argued
that anti-yellow fever campaigns had made transmission impossible
in the towns and that yellow fever had therefore become a disease
of the bush instead of a town-disease, but even out in the country
non-immune foreigners travel, apparently without ever suffering
from attacks of yellow fever. Besides, cases of * vomiting sickness ’
occurred, in my experience, in Kingston itself, and were
common in the neighbouring district of Lower St. Andrew, which
is much frequented by tourists, and in which many Europeans have
their permanent residences. The seasonal incidence also is almost
destructive to the hypothesis of yellow fever. Yellow fever occurs,
without any doubt, at any time of the year in endemic areas, but it
has never been known to show the exceedingly abnormal feature of
dying out or becoming very rare during the hot and rainy seasons,
when mosquitos, according to local information, are abundant,
whilst, on the other hand, it prevails during the cool and dry
months, during which, in my experience, mosquitos in general are
rare, and S. fasciata very rare. Thus epidemiological evidence is
rather against identifying ‘vomiting sickness’ with yellow fever;
this should, therefore, only be done if the clinical and pathological
evidence were absolutely overwhelming. But this is by no means so.
The classical clinical symptoms of fatal yellow fever—as
fever, black vomit, jaundice and anuria—are almost constantly
absent. They may be absent, one or all, in cases of yellow
fever, but it is hardly conceivable that they should all
be absent in practically all cases in large outbreaks.
457
Fulminant yellow fever has been described, but not of the type
which is met with in ‘ vomiting sickness.’ The anatomical
lesions in many cases of ‘ vomiting sickness' are certainly closely
similar to those observed in yellow fever, but, then, I have
previously (1911, 1) maintained that the anatomical diagnosis of
yellow fever is even more difficult than the clinical one. No single
pathological phenomenon is pathognomonic in yellow fever, but the
combined picture of clinical symptoms and anatomical lesions is
nearly always so. This combined picture, however, has not been
met with in ‘ vomiting sickness.’ With regard to the most important
lesions it may be emphasized that haemorrhagic gastritis was
common, but by no means constant in ‘vomiting sickness,’ and
rarely as marked as in yellow fever; the affections of kidney and
especially liver differed in showing more marked fatty changes and
less marked necrobiotic phenomena than in yellow fever, especially
when the comparatively slowly developing cases of ‘ vomiting
sickness ’ are considered, as they naturally must be very particularly,
as one would expect to find in such cases the most typical lesions.
The Rocha-Lima type (see note on p. 438) of liver affection has
not been marked in any case of ‘ vomiting sickness,’ and, as a rule,
not even a similar type has been observed.
The most striking feature of all is, perhaps, that these abnormal
epidemiological, clinical and pathological characters, if the disease
really were yellow fever, should have repeated themselves year after
year in an absolutely typical manner. This certainly is a very
strong argument which tends to show that we have to do with a
specific and typical disease.
As regards parasitological evidence, little can at present be said.
I had very rarely the opportunity of examining good blood smears
taken whilst the patient was alive. In no case of typical ‘ vomiting
sickness,’ however, was the Paraplasma flavigenum found. I shall
not here discuss the importance of the parasitological evidence in
yellow fever, but must refer to my previous publications (especially
1911, 2 and 1912). The one case (Case 23) in which P. flavigenum
was found, but which was not typical ‘ vomiting sickness,’ has been
discussed in the preceding section and will again be referred to in
Section VIII.
The other disease with which ‘ vomiting sickness ’ has been, at
458
least partially, identified is cerebro-spinal meningitis. The
evidence is that meningococci have been found by Scott (1912,
1 and 2, 1913, 1 and 2) in the cerebro-spinal fluid in a considerable
number of cases of 1 vomiting sickness.* Scott himself is, however,
in his latest paper less positive with regard to the essential
importance of his results, as far as 4 vomiting sickness * is concerned,
than he was to begin with. At any rate, there is more evidence in
favour of the identity of 1 vomiting sickness * with meningitis than
with any other disease, and the question must be most seriously
considered.
In the first place the bacteriological findings must be discussed.
I shall not discuss the results published by Scott, but limit my
remarks to my own investigations, in which I was, as stated above,
greatly assisted by Dr. Scott himself. We fully realized—at least
it soon became evident—the great difficulty in obtaining an
absolutely reliable bacteriological diagnosis of meningococci, a
difficulty which is, I believe, greater in Jamaica and probably in
other tropical countries than elsewhere, as I shall presently explain.
We therefore resolved to use the agglutination test, and not being
in possession of a meningococcus strain from another locality used a
strain obtained in one of Dr. Scott’s first cases for immunization
purposes. Unfortunately, however, the time at my disposal was
too short; in fact, I gave the last of the commenced series of
injections to several rabbits undergoing immunization, just before
I left Jamaica. Thus I had to rely upon the morphological and
cultural characters, staining reactions, and sugar reactions of the
diplococci found. Of these characteristics, only the sugar reactions
need a few preliminary remarks, as we, of course, only considered
a diagnosis of meningococcus when colonies developed on
ascites-agar, consisting of coffee grain-shaped diplococci, which
did not retain the colour in the Gram-process. As regards
the production of acid in sugar-media, there is no perfect
agieement even between the most recent authors on the subject.
Busse (1910) and Kutscher (1912) adopt Lingelsheim’s view that
meningcocci change glucose and maltose, but not other sugars,
whilst Netter and Debre (1911), though in a general way confirming
the same experiences, state that no very severe rules should be
laid down. Scott (1913, 2) isolated in his first cases a diplococcus
459
which changed both glucose, maltose and galactose, leaving other
sugars unchanged. These cocci were otherwise typical, and they
were found in cases of anatomically well-characterized meningitis.
For this reason we adopted these characters as a preliminary basis
for classification, and according to this classification the distinction
is made between the Groups A and B of my cases. The twelve
strains isolated in the cases belonging to Group B showed the
following reactions: —
No.
Glucose
Maltose
Galactose
Lactose
Saccharose
Mannite
1
+
4 -
4 -
_
-
7
+
4 -
-
-
-
8
4 -
4 -
4 -
-
-
-
12
+
4 -
4 -
-
-
-
20
4 *
4 -
—
-
-
-
30
4-
4 -
+
-
i
-
33
4-
4 -
+
-
' 1
-
43
+
4 -
-
-
_
-
44
4 -
4 -
4-
-
-
-
50
4 -
4 -
4-
-
-
-
55
+
4 -
4-
-
1
-
57
+
4 -
4 -
-
! " 1
1
Thus according to the classical rule, only the strains in Cases 7,
20 and 43 should represent typical meningococci. These three
cases showed no more pronounced meningeal lesions than the others.
In Group D, cases of meningitis, anatomically well characterized,
the following reactions were obtained.
No. |
!
Glucose
1
Maltose
Galactose
Lactose
Saccharose
Mannite
4
4-
I
4*
-
-
-
19
4*
4-
4*
-
-
-
29
4-
4-
4-
-
-
-
31
+
+
4*
-
-
-
39
4-
4-
4-
—
—
—
460
In view of these results in the cases in Group D it appeared
natural to uphold the distinction as previously stated.
No further unusual characters were observed during the examina¬
tion of the cultures in Jamaica, but they became evident on
re-examination of the cultures in Liverpool, where I continued
several strains in Professor Beattie’s laboratory. Already the
survival of the cultures was remarkable. Some of them, which
I brought home myself, survived twenty-seven days until sub¬
cultured. Other cultures, which Dr. Scott sent me, lasted for over
two weeks. The following strains were examined: Cases 1
and 4 (A and B) and two of Dr. Scott’s cultures, Stock I and
Stock II. The following reactions were obtained: —
No.
Glucose
Galactose
Lactose
Saccharose
Mannite
. (B.D.)
! +
—
—
-
4 (V.C.) A
+
-
-
-
-
4 (V.C.) B
+
+
4-
+
-
Stock I
+
-
-
-
-
Stock II
+
i
+
+
1
1 +
-
I now asked Dr. Ledingham, of the Lister Institute, for an
immune serum for agglutination purposes. Unfortunately no
rabbit immune-serum was available, but I received some of the
Institute’s anti-meningococcus horse serum, which Dr. Arkwright
informed me showed marked agglutinating power, having been
tested with numerous meningococcus strains. I likewise received
normal horse serum from the Lister Institute. The results were as
follows (macroscopical methods used): —
No.
Antimeningococcic serum
Normal
hone
serum
1-20
1-40
1-80
1-160
1-320
1-640
1-20
. (B.D.) ...
+ 4- 4-
4-4- 4-
4-
-
-
-
4-4-4-
+ (V.C.) A ...
4-4--f
+ + +
-
-
4- f
4 (V.C.) B ...
+++
4-4-4-
4-4-4-
4-4-
1
4-4- |
1
+
4-4-4-
Stock I
+++
4-4-
4-
-
-
-
4-4-4-
461
Only the strain 4 (V.C.) B gave a marked reaction; this result
is remarkable, as this same strain gave quite atypical sugar reactions.
It is, however, stated by several authors that there is a marked
‘ group reaction ’ amongst the meningococci and allied species. As
the numbers of sub-cultures increased it was also observed that their
aspects gradually changed; the growth was more abundant; and
the colour more saturated whitish, characters which were particularly
marked in Strain I (B.D.). The morphological characters remained
the same, but the staining was somewhat irregular; it was found
that on repeated occasions several of the strains showed a
remarkable resistance to the decolorization after Claudius.
Dr. J. A. Arkwright also examined one of these cultures
[4 (V.C.) B] and kindly informed me that he had observed con¬
siderable differences from the usual meningococcus strains, especially
a considerably higher resistance. He found that the strain produced
acid in glucose, maltose, galactose (slight) and cane sugar, but not
in lactose.
Having learned that these strains possessed a considerable degree
of variability, it appears advisable to revise Group A, as in many of
the cases therein included diplococci were found, which were not
considered in the discussion on meningococci on account of
differences in sugar reaction or other characters. The following
table shows these observations: —
The strain in 37 showed but little difference from those
previously described; in fact, sub-cultures gave no reaction in
lactose media, which would perhaps justify including this case in
Group B. The strains 9 and 46 might be considered typical
462
meningococci (^according to Abel (iqi 1) only glucose or maltose
should be changed by meningococci), were it not for the peculiarity
that the diplococci appeared in well marked chains. In this respect
it resembles a strain which I found in a case of purulent meningitis
observed in Yucatan (1910), and in which the formation of chains
was a salient feature.
No pathogenic power was found by subcutaneous, intravenous,
intraperitoneal and intracardial inoculations of living cultures of
several strains into guinea-pigs or rabbits. These experiments need
therefore not be detailed.
Thus, the bacteriological examination did not solve the question
in an indisputable manner. The nature of the diplococci observed
is not beyond question; but one thing seems evident; the distinction
between Groups A and B, cases of ‘ vomiting sickness * type without
and with meningococci, which was already at first but poorly
marked, has been still more weakened by the closer examination of
the cultures.
From the bacteriological data it is impossible to say whether
the organisms isolated have any pathogenic importance or not.
The organisms are not typical meningococci, but it is quite possible
that they may cause pathological phenomena. I have, so far, come
to conclusions similar to those of Arkwright (1909), who says about
several organisms examined, ‘ They apparently have no relation¬
ship to the Meningococcus of Weichselbaum. They may, however,
be the causal organism in some cases of meningitis, or may play a
secondary part/
The question, if possible to decide at the present time, must be
decided on clinical, anatomical and epidemiological grounds.
Clinically the typical cases of * vomiting sickness * show consider¬
able resemblance to fulminant cases of cerebro-spinal meningitis.
Such cases have been described by many authors and are mentioned
in recent text books. Thus, Osier (1912) says, ‘Malignant form:
This fulminant or apoplectic type is found with variable frequency
in epidemics. It may occur sporadically. The onset is sudden,
usually with violent chills, headache, somnolence, spasms in the
muscles, great depression, moderate elevation of temperature, and
feeble pulse, which may fall to fifty or sixty in the minute.
Usually a purpuric rash develops. In a Philadelphia case, in 1888,
463
a young girl, apparently quite well, died within twenty hours of
this form. There are cases on record in which death has occurred
within a shorter time. Stille tells of a child of five years, in whom
death occurred after an illness of ten hours; and refers to a case
reported by Gordon, in which the entire duration of the illness was
only five hours. Two of Vieusseux’s cases died within twenty-four
hours/
* In malignant cases there may be no characteristic changes, the
brain and spinal cord showing only extreme congestion, which was
the lesion described by Vieusseux/
Koplik (1907) mentions a malignant form of meningitis which
is fatal within twelve, twenty-four or thirty-six hours. 1 An adult
or a child in apparent health complains of slight headache, there
is nausea followed by vomiting, fever and unconsciousness in rapid
succession. Death occurs in a short time, consciousness not being
regained/
A good description is also given by Ormerod (1905). This
hyperacute form is particularly common in children, and is
described in pediatric text books, for instance, Cautley (1910),
Thiemich (1910) and Holt (1913). The latter says 'Cases of this
kind are rarely seen except in an epidemic, and usually occur at
its height. The onset is very abrupt, the course short and intense,
and death may take place in from twelve to thirty-six hours.
The following case illustrates this type: A little girl of ten years
was well enough at 2 p.m. to carry a bundle of cloths a dozen
city blocks. Returning home, she complained of intense headache,
vomited frequently, and was so weak that she was obliged to go to
bed. In a few hours she passed into deep coma, with very high
fever and died at 11 p.m/ (p. 704.)
‘ There may be only a serous exudation and intense hyperaemia,
which is doubtless much less marked after death than during life.
The cerebro-spinal fluid is turbid and much increased in amount/
The absence of definite inflammatory lesions of the meninges is
recognized by the above authors and especially described by Busse
(1910) and others. The whole question is particularly well dealt
with by Netter and Debre (1911). Thus there is no doubt that such
cases have been described, and so far there is no objection why the
cases should not be diagnosed as hyperacute forms of meningitis.
464
With regard to the anatomical lesions of other organs than the
meninges it may be mentioned that Busse (1910) lays stress upon the
affection of the lymph-nodules, both peripheral and mesenteric.
This corresponds to the conditions met with in vomiting sickness.
On the other hand, such severe affections of liver, pancreas, and
kidneys, as those found in my cases, have, as far as I know, not
been described in cases of meningitis.
The typical case of ‘vomiting sickness’ corresponds fairly well
to descriptions given of hyperacute cases of meningitis. It differs,
however, as far as our present knowledge goes, in one important
particular, namely, in the absence of fever. In the fulminant form
of meningitis it is the rule that the temperature rises up rapidly,
often to about 105°, or even more, whilst it is only mentioned as
quite an exceptional thing that the temperature occasionally may
remain normal until the fatal termination.
The epidemiological conditions, however, must also be
considered. With regard to this question, practically all authors
state that fulminant cases occur incidentally, isolated during
epidemic outbreaks, and it is said by several, that they occur chiefly
at the height of epidemics. The conditions in Jamaica are quite
different. Here we have recurrent yearly outbreaks, and, perhaps,
smouldering endemic prevalence.
It seems quite evident that no similar outbreaks of meningitis
have ever been described. I therefore cannot accept at the present
state of the investigations, the explanation, which at first sight
seems the most probable, that ‘ vomiting sickness ’ is simply a form
of meningitis. Experience has shown in several diseases that a
germ which has for some time been regarded as a pathogenic one,
has, on close investigation, been reduced to a secondary position as
representing only a complicating infection. Further investigation
is therefore absolutely imperative before the problem can be con¬
sidered finally solved. It is quite possible that ‘ vomiting sickness ’
may be due to a blood-inhabiting, presumably protozoal, organism,
and that a diplococcus infection of the meninges occurs as a
frequent complication/ as a rule not giving rise to any marked
anatomical lesions, because of the rapidly following death, but in
a few cases producing a typical fibrino-purulent meningitis.
Another possibility is that the causal organism may be parasitic in
4 6 5
the intestine and produce exceedingly active toxins. Both explana¬
tions would correspond equally well to the clinical symptoms and
both might also explain the anatomical lesions. The similarity
of the latter to those observed in yellow fever makes one consider
the existence of a blood parasite as very probable, but the marked
affection of the lymph nodules in various parts of the body would,
perhaps, be more in favour of a bacterial infection. In either case,
whether the diplococci observed in these cases are the essential
causal organisms or only of secondary importance, it seems, at any
rate, very unlikely that they should be of no pathogenic importance
at all. Whether these diplococci represent one or more species, it is
impossible to say. At any rate they differ obviously from meningo¬
cocci, gonococci, Diplococcus crassus and other known diplococci.
The existence of more than one species is suggested by
the very considerable differences in sugar reactions and
agglutinating power. On the other hand, the great variability
observed in some of the strains makes it possible to assume, until
further investigation, the existence of only one new species. For
this new species, which is very likely to be only a provisional one,
I would suggest the name Diplococcus jamaicensts .
The chief characteristics of this species are that it is a diplococcus
morphologically similar to gonococcus and meningococcus, as a rule
Gram-negative, but occasionally decolourizing with some difficulty,
showing fairly abundant growth on nasgar, the cultures being more
resistant than meningococcus cultures, and changing at least glucose
or maltose, but, as a rule, often one or several other sugars.
The mode of transmission of ‘ vomiting sickness * I have had no
opportunity of investigating. As far as possible I examined the
prevalence of mosquitos, ticks and other insects in houses which
I visited with the object of seeing cases. Mosquitos were not
commonly found, and when found were not abundant. Some were
Stegomyia fas data, some Culex sp.
I have not examined systematically cultures from the naso¬
pharynx of contacts with ‘vomiting sickness’ cases, and I do not
anticipate that very definite results can be obtained in this way until
the etiology of ‘vomiting sickness’ is definitely settled. At
present one would be in great difficulty with regard to a
diagnosis between atypical strains of Diplococcus jamaicensis and
4 66
D. catarrhaliSy and, further, one would not know how much
importance should be attached to the positive results of the examina¬
tion for Diplococcus jamaicensis.
Other theories, such as vomiting sickness being identical with
malaria, with some form of helminthiasis, or with some kind of
poisoning, find no support whatever in my observations.
VII. PROPHYLACTIC MEASURES
The absolute lack of knowledge about the mode of transmission
of ‘ vomiting sickness 1 makes it impossible to lay down any rules
with regard to prophylactic measures. On general hygienic
principles it may be recommended that the poverty should be
relieved and popular education, especially with regard to cleanli¬
ness, promoted. It is, of course, also important that anti¬
mosquito campaigns should be carried out, as well as campaigns
against other insects, but to none of these measures can any
specific importance be attached. It has been stated that no evidence
points to insect transmission of the disease, and with regard to
poor hygienic conditions it has also been stated previously that a
considerable number of the individuals attacked by ‘ vomiting
sickness* were absolutely healthy looking and well nourished;
sometimes the houses and huts visited were also well kept and
clean.
VIII. OTHER OBSERVATIONS IN JAMAICA
The one observation of a yellow fever case which has been
mentioned in Section IV, would apparently tend to show that
other cases might be of the same nature. There is, however, no
evidence which makes it necessary to adopt this view. The cases
occurred at the same time and in the same place as several cases of
‘ vomiting sickness/ but it showed a clinical course different from
those cases, and the anatomical lesions were also different. There
is, of course, no reason why the two diseases should not occur at
the same time and in the same place, supposing that both exist in
the Island. The problem is how to explain the occurrence of one
467
isolated case of yellow fever. This, however, is only what
happens in many endemically but mildly infected places where
the existence of yellow fever only becomes known when there is
a considerable influx of foreigners. That outbreaks of yellow
fever do not occur in Jamaica, though the disease may exist
amongst the natives, is easily explained, I believe, by the relative
freedom from mosquitos which the larger towns enjoy. The
principal travelling public, the tourists, visit Jamaica during the
winter months only, and at this time of the year I can safely assert
that Stegomyia fas data is not common in parts of Jamaica which
I have visited, and which I believe in their natural conditions
correspond fairly well with the various places which foreigners
would visit.
I shall not mention here the laboratory observations on other
subjects which were made during the ‘ vomiting sickness * investiga¬
tion, but I want briefly to refer to a case which presented consider¬
able interest, especially when considered in connection with my
observations on Cutaneous Leishmaniasis in Yucatan (1912). This
was a case of a fistulous and ulcerated tumefaction of the lower
part of the femur, in which case I suspected infection with
Leishmania tropica , but this parasite was never found, though
repeatedly examined for. The particularly interesting point is
that the exudation contained numerous intracellular diplococci,
similar to those which were a constant feature in the cases of ear
ulcer in Yucatdn. Cultures were not obtained of this diplococci
because of abundant mixed infection.
IX. OTHER PLACES VISITED
On my way to Jamaica and back I had the opportunity of
visiting Trinidad, Barbados, the Panama Canal Zone, and
Colombia. I visited the hospitals and saw interesting cases in
Barbados and Trinadad and was fortunate in having a very good
opportunity of seeing the sanitary arrangements on the Panama
Canal Zone.
I also had the opportunity of discussing sanitary matters with
Colonel Gorgas, Dr. Perry and other Officers of the Health
Department, and was much impressed by the thoroughly well-
established organisation.
468
X. ACKNOWLEDGMENTS
The thanks of the Liverpool School of Tropical Medicine are
due to those friends who generously provided the funds for the
‘ Vomiting Sickness ’ Expedition, to the Royal Mail Steam Packet
Co. for granting free passages to and from Jamaica, as well as to
the Leyland Line and Messrs. Elder & Fyffes for the free transport
of goods.
I wish to express my gratitude to His Excellency Sir Sidney
Olivier, Governor of Jamaica; His Excellency Mr. P. C. Cork,
Acting Governor, and His Excellency Sir William Manning,
Governor of Jamaica, who kindly arranged that all the facilities of
the Health Department should be given to me for the furtherance
of my investigations.
I am exceedingly indebted to the Hon. J. Erington Ker,
Superintending Medical Officer, for the excellent arrangements
which he made for reporting cases of vomiting sickness and for help
in many other ways, as well as for great hospitality.
I am also greatly indebted to Dr. H. H. Scott, Government
Bacteriologist, in whose laboratory I worked and who helped me in
every possible way, as stated above.
I also wish to express my thanks to the following District
Medical Officers:—Dr. Clarke, Dr. Cooke, Dr. Curphey,
Dr. Earle, Dr. Edwards, Dr. George, Dr. Gifford, Dr. Joslen,
Dr. Peck, Dr. G. W. Thomson, as well as to the Medical Officers
of the Public Hospitals in Kingston, Dr. Castle, Dr. Ross,
Dr. C. A. H. Thomson and Dr. Dryden, also to Dr. MacDonald
(City Health Officer), Dr. Crosswell and Dr. Saunders.
K-i dney*
Text
469
REFERENCES
Abel, R (1910). Baktcriologisches Taschcnbuch. 14th Ed.
Annual Report of the Superintending Medical Officer, etc., Kingston, Jamaica, 1903, 1905,
1906, 1907, 1908, 1909, 1910, 1911, 1912.
Arkwright, J. A. (1909). Varieties of the meningococcus with special reference to a com¬
parison of strains from epidemic and sporadic sources. Journ. Hyg., Cambridge, Apr.,
IX, 1, pp. 104-121.
Branch, C. W. (1906). Vomiting sickness of Jamaica. Journ. Trop. Med. Hyg., London,
Dec. 15, pp. 374 - 375 -
Bussx, O. (1910). Die ftbertragbare Genickstarre. Jena. (Reprint from Klin. Jahrb.,
XXIII.)
Cautliy, E. (1910). The diseases of infants and children. London. (Cerebrospinal
meningitis, pp. 647-660.)
Holt, L. E. (1912). The diseases of infancy and childhood. New York and London. 6th
Ed. (Cerebrospinal meningitis, pp. 701-717.)
Kxr, J. E. Vide Annual Report.
Koplie, H. (1907). Epidemic Cerebrospinal meningitis. In Osier and McCrae : A System
of Medicine. London. II, pp. 496-520.
Kutscher, H. H. (1912). Uebertragbare Genickstarre. In Kolle & Wassermann : Hand-
buch der pathogenen Mikroorganismen, 2nd Ed., IV, pp. 589-654.
MacDonald, A. (1913). Is Yellow Fever endemic in Jamaica? (A paper read before the
Jamaica Branch of the British Medical Association, Dec. 1912.) Pp. 17. 8°.
Kingston.
Netter, A., & Debre, R. (1911). La m£ningite c6r6bro-spinale. Paris. Masson et Cie.
Ormerod, J. A. (1905). Cerebrospinal fever. In Allbutt and Rolleston : A System of
Medicine. London. I, pp. 923-942.
Osler, Sir W. (1912). The Principles and Practice of Medicine. 8th Ed. New York and
London. (Cerebro-spinal fever, pp. 108-115.)
Potter, T. J. (1912). Vomiting Sickness in Jamaica. Report to the Colonial Office. Suppl.
Jamaica Gaz., Nov. 23, XXXV, 24, pp. 695-715.
Rocha-Lima, H. da (1912). Zur pathologischen Anatomie des Gelbfiebere. Verhandl.
Deutsch. Pathol. Gesellsch., Apr. 15-17, pp. 163-182.
SchottmOller, H. (1910). Pathymeningitis interna infectiosa acuta u. Meningismus.
Munch. Med. Woch., LVII, 38, pp. 1984-86.
Schultze, F. (1901). Die Krankheiten der Hirnhaute und die Hydrocephalie. Nothnagel’s
Spec. Path. u. Therap., Ill, 3, pp. 1-258.
Scott, H. H. (1912, 1). Third Six-Monthly Report on the Work of the Government Bacteri¬
ologist (Kingston, Jamaica). London.
- (1912, 2). Interim Report on 4 Some cases of Vomiting Sickness at P . . .
Dec. 25.
- (i9 i 3j 0* The similarity between the symptom-complex of cerebrospinal meningitis
and some cases of vomiting sickness, Feb. 7.
-(1913,2). Fulminating Cerebro-spinal Meningitis in Jamaica. Ann. Trop. Med.
Paras., Liverpool, VII, 1, pp. 165-181.
Seidelin, H. (1910). Experiences in Yucatan. Journ. Trop. Med. Hyg., Lond., Nov. 15,
xiii, 22, pp. 335-340.
-(1911, 1). The Post-mortem diagnosis of yellow fever. Y.F. Bur. Bull., Liverpool,
Sept., I, 5, pp. 173-192.
-(1911 9 2). The etiology of yellow fever. Y.F. Bur. Bull., Liverpool, I, 7, pp. 229-258.
-(1912). Report of the Yellow Fever Expedition to Yucatan, 1911-1912. Y.F. Bur.
Bull., Liverpool, Oct., II, 2, pp. 123-242.
Thiemich, M. (1910). Meningokokkenmeningitis. In Pfaundler & Schlossmann : Hand-
buch der Kinderheilkunde, Leipzig, IV, pp. 408-422.
Turton, R. S. (1904). The * vomiting sickness ’ of Jamaica. Journ. Trop. Med. Hyg., London,
June 1, VII, pp. 163-164.
470
EXPLANATION OF PLATES
Plate XXIX
Figs. 1-2. Two huts in which cases of vomiting sickness occurred.
These figures show the very primitive structure and poor
appearance of man)' of the native huts and their
situation in the bush.
Annals Trop. Med. & Parasitol., Vol. VI1
PLATE XXIX
Fig. 2
C. Tinting 6* Co., Ltd., Imp.
+72
Plate XXX
All figures are from safranin-stained sections of specimens fixed
in Fleming’s strong osmic acid solution, and all are drawn by means
of Abbe’s drawing apparatus, the optic system being Zeiss’s apoch.
4 mm. and compensating ocular 4. x 360. All figures on this and
the following plate have been drawn by Mrs. Margrethe Seidelin.
Fig. 1. Stomach. Marked fatty change of epithelial cells.
Fig. 2. Pancreas. Marked fatty metamorphosis; small and
medium-sized fat droplets are present in practically all
the epithelial cells of the alveoli. A Langerhan’s islet
is shown in the figure, but it is not very sharply defined;
its cells have undergone fatty change, but less marked
than in the case of the alveoli. The epithelia of the
excretory ducts are in particular severely affected.
Fig. 3. Myocardium. Moderate fatty metamorphosis.
Fig. 4. Liver. Intense fatty metamorphosis.
Annals Trop. Med. ParasitolVol. Vll
PLATE XX J
474
Plate XXXI
Technique as indicated for Plate XXX (x 360):
Fig. 1. Suprarenal capsule (Case 55) cortex. A considerable
amount of fat is present in nearly all epithelial cells.
Fig. 2. Kidney (Case 62). Moderate fatty change, both convo¬
luted tubules and glomeruli cells being affected, the latter
less than the former, but in the case of both only minute
fat droplets are observed in the cells.
Fig. 3. Kidney (Case 47). Intense fatty metamorphosis, the
epithelia of the convoluted tubules contains very large fat
drops; in the lining epithelia of Bowman’s capsules and
the glomeruli cells fatty change is also observed, but
much less marked.
Fig. 4. Kidney (Case 47). Intense fatty metamorphosis of the
epithelia of the ascending limbs of Henle’s loops,
whilst in the descending limbs the change is very
little marked, only some of the epithelial cells showing
minute fat droplets.
Annals T Top. Med. & Parasitol., Vol. VII
PLATE XXXI
%
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9 *£"
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474
Plate XXXI
Technique as indicated for Plate XXX ( x 360):
Fig. 1. Suprarenal capsule (Case 55) cortex. A considerable
amount of fat is present in nearly all epithelial cells.
Fig. 2. Kidney (Case 62). Moderate fatty change, both convo¬
luted tubules and glomeruli cells being affected, the latter
less than the former, but in the case of both only minute
fat droplets are observed in the cells.
Fig. 3. Kidney (Case 47). Intense fatty metamorphosis, the
epithelia of the convoluted tubules contains very large fat
drops; in the lining epithelia of Bowman's capsules and
the glomeruli cells fatty change is also observed, but
much less marked.
Fig. 4. Kidney (Case 47). Intense fatty metamorphosis of the
epithelia of the ascending limbs of Henle’s loops,
whilst in the descending limbs the change is very
little marked, only some of the epithelial cells showing
minute fat droplets.
476
Plate XXXII
MICROPHOTOGRAPHS
Fig. i. Mucosa of stomach (Case 55), showing hyperaemia, diffuse
lymphoid infiltration, and superficial necrosis and
haemorrhages. To the right, part of a lymphoid follicle
is seen, x 95.
Fig. 2. Mucosa of duodenum (Case 34), showing hyperplasia of
Brunner’s glands and diffuse microcellular infiltration;
also superficial necrosis. Remnants of the muscularis
mucosae are seen, but this coat has been entirely broken
up by the proliferated glands, x 95.
Fig. 3. Liver (Case 55), showing intense diffuse fatty meta¬
morphosis and moderate necrobiotic changes, likewise of
a diffuse character (x 370). Other parts of the same
liver showed intense capillary hyperaemia.
Pancreas (Case 43), showing marked necrobiotic changes,
x 370.
Fig. 4.
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479
STUDIES IN BLACKWATER FEVER
(i) STATISTICAL
BY
J. W. W. STEPHENS, M.D. (Cantab.)
In this paper I propose to consider some of the facts bearing on
the malarial origin of blackwater fever which I have been able to
collect from an examination of recent literature.
Arguments of a general nature, though there are many, I shall
not consider here, but shall examine the facts so far as I have been
able to ascertain them under the following headings: —
(1) Malaria parasites.
(2) Pigmented leucocytes.
(3) Post-mortem examinations.
(4) Influence of malaria.
(5) Relationship to species of malaria parasite.
(6) Effect of period of residence.
(7) Seasonal prevalence.
(8) Correlation between malaria and blackwater statistics.
(9) Second attacks.
In 1901, Christophers and myself (1901), as a result of our
microscopical studies of blackwater fever, came definitely to the
conclusion that blackwater is malarial in origin but is not simply
an attack of malaria, but occurs only in those who are in a condition
induced by repeated malarial infection lasting over a certain time.
MALARIA PARASITES
One of the most important facts, in my opinion, bearing on the
aetiology of blackwater fever is the presence of malaria parasites in
the blood of blackwater cases. In the literature I have been able
to collect 390 cases. I divide them into three categories, those on
the day before, those on the day of, and those on the day after
the onset of blackwater, as in the accompanying Table I. My
reason for doing this is that in my experience, and also in that of
others, parasites when found rapidly disappear.
Kleine (1901)
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No. 4
ANNALS
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Edited by
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Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
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School of Tropical Medicine ,
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482
This means that in 100 cases of blackwater examined on the
day before the onset, parasites will be found in 73 cases, in
100 cases examined on the day of onset in 47*5* and in 100 cases
examined on the day after in 23.+
An important deduction that I think may safely be made from
these figures is that parasites disappear, and indeed rapidly, during
the course of the disease, as we had observed in our original studies.
This disappearance of parasites is in all probability associated with
the acute haemolysis that is taking place in blackwater fever, and
should be borne in mind in all comparisons between the parasite
rates in blackwater fever and malaria.
Further, the fact that on the day before the onset of the black¬
water parasites are found in 73% of cases seems to point to the
initial attack being closely associated with a malarial attack. That
relapses do occur during the course of the disease (which relapses
in some cases at least can be brought on by quinine) without the
occurrence of parasites in the blood, seems to be certain, though I
have not collected the data on the subject. And even if parasites
were rarely found in blackwater, it would not necessarily exclude
its malarial origin, because it might be that it was just in the
chronic cases of malaria where parasites are absent or scanty that
blackwater occurred, but I will not pursue the argument further,
as I wish as far as possible in this paper to ascertain facts and
avoid discussion.
The following table, compiled from Panse’s (1902) cases, 35 in
number, enables one to show in more detail how time affects the
result of the examination for parasites in blackwater: —
Table I a
Time of blood examination
Cases
Positive
Percentage
Day before onset ...
9
8
88-9
Day of onset ( a ) before onset
11
8
727
„ ( b ) after onset
11
6
54*5
Within 12 hours after onset .
12
6
50*0
More than 12 hours after onset
3 1
9
29*0
• A correction is probably necessary for those cases occurring on the day <?/*the blackwater
for some of these will probably have been examined before the onset of the blackwater and
some (probably the majority) after, so that on the assumption that the parasite rate before
the onset and that of the day before is the same, this figure should be reduced, but it is
impossible to say by how much.
t Or put in a different way of ioo positive cases. 51 will occur on the day before,
32-8 on the day of, and 16 on the day after the blackwater.
479
STUDIES IN BLACKWATER FEVER
(i) STATISTICAL
BY
J. W. W. STEPHENS, M.D. (Cantab.)
In this paper I propose to consider some of the facts bearing on
the malarial origin of blackwater fever which I have been able to
collect from an examination of recent literature.
Arguments of a general nature, though there are many, I shall
not consider here, but shall examine the facts so far as I have been
able to ascertain them under the following headings: —
(1) Malaria parasites.
(2) Pigmented leucocytes.
(3) Post-mortem examinations.
(4) Influence of malaria.
(5) Relationship to species of malaria parasite.
(6) Effect of period of residence.
(7) Seasonal prevalence.
(8) Correlation between malaria and blackwater statistics.
(9) Second attacks.
In 1901, Christophers and myself (1901), as a result of our
microscopical studies of blackwater fever, came definitely to the
conclusion that blackwater is malarial in origin but is not simply
an attack of malaria, but occurs only in those who are in a condition
induced by repeated malarial infection lasting over a certain time.
MALARIA PARASITES
One of the most important facts, in my opinion, bearing on the
aetiology of blackwater fever is the presence of malaria parasites in
the blood of blackwater cases. In the literature I have been able
to collect 390 cases. I divide them into three categories, those on
the day before, those on the day of, and those on the day after
the onset of blackwater, as in the accompanying Table I. My
reason for doing this is that in my experience, and also in that of
others, parasites when found rapidly disappear.
+8 4
disease, the lower percentage in blackwater is readily accounted for.
The figure for malaria is probably low, but in the Panama figures
of Deeks and James it should be remembered that only a single
examination was made on admission to hospital.
Table IIIa. —Parasite Rate in Blackwater Fever and Malaria
Blackwater Fever
Cases
Positive
%
Observer
39 °
* 6 3
418
Various observers (Table I)
230
89
387
Deeks & James (1911)
3*3
178
1
465
Lovelace (1913)
1003
43 °
42-9*
Malaria
40,928
23,410
57-2
Deeks & James (1911)
16,434
9.«55
557
Lovelace (1913)
57,361
32,565
56-8
The objection has been brought against figures of this kind that
in a malarial country parasites would be found in the same or about
the same percentage in people not suffering from blackwater fever,
but such objections have not been supported by actual data.
Christophers and myself (1901), moreover, examined 44 Euro¬
peans at Lagos, taken at random. They were especially from
communities suffering much from malaria. We found parasites
in 2, pigmented leucocytes in 2, i.e, 4 infected out of 44, about 9%.
Christophers and Bentley (1908) examined 66 babus in the Duars,
Bengal, and found parasites or pigment in 14, i.e., 212%. In
10 cases of blackwater on the day of onset they found parasites or
pigment in 8, or 80%. Further we have the control figures of
Table II.
This figure represents the positive parasite rate irrespective of the day of the disease and
should be compared with the figures of Tables I and II,
4»5
If, then, these data are correct, they imply that in the general
population and those suffering from diseases other than malaria or
blackwater fever the parasite rate is not comparable with that in
blackwater fever.
Moreover, I believe we are justified in meeting the above
objection more forcibly still. We have in blackwater a disease in
which immediately before the onset of the disease malaria parasites
are found in 73% of cases, taking the figure of Table I. If this
does not signify that the disease is malarial, then the finding of
parasites in 73% of cases of ‘ malaria * has also no significance, and
‘ malaria * is also due to some unknown cause.
PIGMENTED LEUCOCYTES
In blackwater fever it is the general experience that parasites
when found rapidly disappear. We still, however, have a means
of detecting a malarial infection if present, viz., by means of
pigmented leucocytes. In the literature most frequently no state¬
ment is made with regard to pigmented leucocytes. The following
figures of Christophers and Bentley are small and only suggestive,
and more observations are required.
Table IV.*—Pigmented Leucocytes in Blackwater Fever (Christophers & Bentley, 1908).
First
Day
Second
Day
Fourth-Sixth Day
No. of
No.
No. of
No.
No. of
No.
cases
positive
cases
'
positive
cases
positive
Parasites .
10
1
6
7 !
0
10
1
Pigment .
1
1 10
1
8
1
7
1
6
10
4
POST-MORTEM EXAMINATIONS
In 31 cases where definite statements positive or negative have
been made malaria pigment was found P.M. in 26, i.e., 83 0%.
The evidence of malaria then in 5 cases is negative. If the absence
of pigment P.M., supposing the facts correct, definitely excludes
malaria then some blackwater cases must be due to other causes,
which one must admit is not impossible, although I believe the
facts prove that malaria is the dominant factor.
* In this table, the data are obtained from the same 10 cases.
482
This means that in 100 cases of blackwater examined on the
day before the onset, parasites will be found in 73 cases, in
100 cases examined on the day of onset in 47*5* and in 100 cases
examined on the day after in 23.+
An important deduction that I think may safely be made from
these figures is that parasites disappear, and indeed rapidly, during
the course of the disease, as we had observed in our original studies.
This disappearance of parasites is in all probability associated with
the acute haemolysis that is taking place in blackwater fever, and
should be borne in mind in all comparisons between the parasite
rates in blackwater fever and malaria.
Further, the fact that on the day before the onse\ of the black¬
water parasites are found in 73% of cases seems to point to the
initial attack being closely associated with a malarial attack. That
relapses do occur during the course of the disease (which relapses
in some cases at least can be brought on by quinine) without the
occurrence of parasites in the blood, seems to be certain, though I
have not collected the data on the subject. And even if parasites
were rarely found in blackwater, it would not necessarily exclude
its malarial origin, because it might be that it was just in the
chronic cases of malaria where parasites are absent or scanty that
blackwater occurred, but I will not pursue the argument further,
as I wish as far as possible in this paper to ascertain facts and
avoid discussion.
The following table, compiled from Panse’s (1902) cases, 35 in
number, enables one to show in more detail how time affects the
result of the examination for parasites in blackwater: —
Table Ia
Time of blood examination
Cases
Positive
Percentage
Day before onset ...
9
8
88-9
Day of onset (a) before onset
11
8
727
(b) after onset
11
6
54*5
Within 12 hours after onset
12
6
50*0
More than 12 hours after onset
1
3 *
9
29*0
• A correction i> probably necessary for those cases occurring on theday of the blackwater
for some of these will probably hare been examined before the onset of the blackwater and
some (probably the majority) after, so that on the assumption that the parasite rate before
the onset and that of the day before is the same, this figure should be reduced, but it is
impossible to say by how much.
f Or put in a different way of ioo positive cases, 51 will occur ou the day before,
32*8 on the day of, and 16 on the day after the blackwater.
483
The next tables, Table II and III, I give separately, as the data
were compiled in a different way to that in Table I.
Table II.— Parasites in Blackwater Fever. Compiled from Deeks and James (1911)
75
cases
examined 4 — 1 days before onset
... Pos.
48
=
64-0%
28
15
,, 1 day before ,,
11
H
-
5 °'°%
36
>1
„ day of onset ...
„
12
-
33-3 %
62
,,
„ 1 — 4 days after onset
... ,,
12
=
' 9 - 4 %
,928
11
of Malaria examined on admission
... ,,
2 3 i 4 10
=
5 7' 2 %
260
11
Amoebic dysentery.
... ,,
48
=
■ 8 - 5 %
70
„
Liver abscess .
... ,,
7
=
10-0%
33
11
Typhoid .
... ,,
4
=
12-1%
_ ?
51
Tuberculosis
About
12-0%
11
Pneumonia.
Less than
12-0%
Table III
Madeira-Mamore Railway Co., Porto Velho, Brazil. Compiled from Lovelace (1911)
Total
blood-examinations
(26 months)
Positive
Rate
Malaria .
16,434
9,‘55
557%
Blackwater*
383
178
46-5%
The tables are also of value because we are able to compare the
percentages in blackwater with those made in malaria under the
same conditions of observer, staining, and probably of time devoted
to each examination.
In the next table, Table IIIa, the data of the first three tables
are summarized.
From it we see that parasites are found in blackwater fever in
42*9% of cases (taking no account of the day of the disease on
which the examination is made), while in malaria, parasites are
found in 56*8% of cases. If we accept the conclusion drawn from
Table I, viz., that parasites disappear during the course of the
• In the majority of cases only one blood examination was made, and it was made during
the period of haemoglobinuria.
488
INFLUENCE OF PERIOD OF RESIDENCE
There is a general impression that blackwater is most prevalent
in the second or third year of residence, but the statistics on this
point are nearly all subject to the objection that no account has
been taken of the number of people in each residential period, first,
second, third, etc., years, and this may vary much from year to
year. Here again, what we require to know in the first place is not
out of one hundred cases of blackwater how many occur in the
first, second, etc., years, but how many cases of blackwater occur
in each one hundred people or similar sample of the different
periods of residence, one, two, three, etc., years. Knowing this,
we can then calculate how many out of one hundred cases of black¬
water occur in the first, second, third, etc., years. Plehn (1901) is
the only observer who supplies us with what we require to know,
though, unfortunately, his figures extend only to two years.
The figures in the second row of Table VIII indicate the number
of people in their first, second, third, etc., month of residence under
observation (for Hgb. estimations). Thus in the second month of
residence there were seventy-five, therefore eight of these were not
under observation in their first month, but whether the remaining
sixty-seven were the same people as the sixty-seven in their first
month, the table does not tell us; probably they were, and at the
end of twenty-four months they were reduced to five, but it is not a
material point.
Tabll VIII.—Blackwater Fever : Effect of Residence (Plehn, 1901).
First Year
Month...
1
1 ! 2
3
1
4
5
6
7
8
9
10
11
12
Residents .
67 7;
75 !
7 i
67
62
61
!
57
55
50
46
47
Malaria cases ...
0
00
60
42
! 41
43
OO
44
37
33
26
*4
B.W. cases
0 0
1
0
0
1
0
1
3
4
6
3
4
1
1
4*5
If, then, these data are correct, they imply that in the general
population and those suffering from diseases other than malaria or
blackwater fever the parasite rate is not comparable with that in
blackwater fever.
Moreover, I believe we are justified in meeting the above
objection more forcibly still. We have in blackwater a disease in
which immediately before the onset of the disease malaria parasites
are found in 73% of cases, taking the figure of Table I. If this
does not signify that the disease is malarial, then the finding of
parasites in 73% of cases of ‘ malaria 1 has also no significance, and
‘ malaria * is also due to some unknown cause.
PIGMENTED LEUCOCYTES
In blackwater fever it is the general experience that parasites
when found rapidly disappear. We still, however, have a means
of detecting a malarial infection if present, viz., by means of
pigmented leucocytes. In the literature most frequently no state¬
ment is made with regard to pigmented leucocytes. The following
figures of Christophers and Bentley are small and only suggestive,
and more observations are required.
Table IV.*—Pigmented Leucocyte* in Blackwater Fever (Christophers & Bentley, 1908).
:
First Day
Second Day
Fourth-Sixth Day
No. of
cases
No.
positive
No. of
cases
No.
positive
No. of
cases
No.
positive
Parasite*
,0
6
7 ! 0
10
,
Pigment
10
1
8
7
' 6
i
10
4
POST-MORTEM EXAMINATIONS
In 31 cases where definite statements positive or negative have
been made malaria pigment was found P.M. in 26, i.e., 83 0%.
The evidence of malaria then in 5 cases is negative. If the absence
of pigment P.M., supposing the facts correct, definitely excludes
malaria then some blackwater cases must be due to other causes,
which one must admit is not impossible, although I believe the
facts prove that malaria is the dominant factor.
* In this tabic, the data are obtained from the same io case?.
486
Table V.—Post-mortem Records of those cates only where definite statement made.
Cases
Parasites
or pigment
positive
Negative
Authority
3
0
3
Barratt & Yorke (1909). But in one of these
scanty pigmented leucocytes, found during
life. '
6
6
0
H. Werner (1907).
*3
13
0
C». H. Whipple (1909).
2
2
0
Christophers & Bentley (1908).
5
5
0
Stephens & Christophers (1901). (B.W. cases
IX-XVI, p. 24.)
2
0
2
Brem (1906). In one case only a smear from
a rib examined.
3 i
26
5
* 3 - 9 %
16*1 %
INFLUENCE OF MALARIA
The influence of malaria in determining blackwater is shown in
a different way by the following table compiled from Deeks and
James.
Table VI.—Influence of Malaria. Ancon Hospital, July, 1904—Sept.. 1910.
Among 40,928
102
Among 42,000
4
cases diagnosed as Malaria,
cases of Blackwater developed subsequent to admission.
cases diagnosed as Typhoid, Pneumonia, Amoebic dysentery, and Tuber¬
culosis, etc. (medical and surgical).
cases of Blackwater developed subsequent to admission.*
RELATIONSHIP TO SPECIES OF MALARIA PARASITE
The statistics on this point are often fallacious. What we
require to know is not the percentage of malignant tertian or other
species of parasite in blackwater cases, but rather what number of
blackwater cases there is in ioo cases of malignant tertian, simple
tertian and quartan respectively. The following figures show this.
* In the surgical wards : 3 after well defined malarial paroxysms, 1 after quinine in a
patient with a history of much malaria.
+»7
Tabli VII.—Compiled from Deeb & James (1911)
Of 23,410
positive
malaria
cases
Of 89 positive
blackwater
cases
Ratio of
blackwater %
to malaria %
Ratio of
M.T. to S.T.
Malignant Tertian .
74%
76-4%
I°3-2%
100
Simple Tertian.
»«%
*3-6%
9°-8%
88
i.e., if x cases of malignant tertian malaria give ioo cases of blackwater.
then a: „ simple „ „ „ 88 „ „
Compiled from Lovelace (1913)
Of 9,155
positive
malaria cases
Of 178
positive
blackwater
cases
Ratio of
blackwater %
to malaria %
Ratio of
M.T. to S.T.
and to
M.T. * S.T.
respectively
Malignant tertian .
5988 = 65*4%
87-48-9%
74 - 8 %
100
Simple tertian.
2760 - 30*1 %
74 - 41 - 6 %
138-2%
185
M.T. + S.T.
39 i - 4 * 3 %
17 - 9 6%
2 * 3 - 3 %
299
Quartan.
16 = 0*2%
O = 0*0
0-0
0
i.e., if x cases of malignant tertian malaria give 100 cases of blackwater.
then x ,,
simple „
n »> *85 ,,
>>
and x „
M.T. + S.T.
» >1 2 99 »
>>
and x „
quartan
»> 0 »>
>>
The discrepancy between these two results I cannot at present
explain, assuming they are both correct, but it should be noted that
Lovelace (1913) in his paper states that the simple tertian parasite
‘ is here an extremely persistent infection. More time and more
quinine are required for its eradication than for that of the aestivo-
autumnal parasite.* And again he speaks of ‘ the rapid blood
destruction in, the great prostration incident to, and the diabolical
persistence of tertian infections.* It would seem as if here we had
an example of increased virulence of the simple tertian parasite.
One*s own clinical experience of the malignant tertian parasite in the
tropics and in temperate zones, seems certainly to point to differences
in virulence.
492
I believe that we may deduce from these figures the fact
that blackwater is not a disease of the first six months or first year,
but mainly of the second year. I believe this fact is capable of
a simple explanation, viz., that blackwater fever only shows itself
in the majority of cases when the patient has been subjected to
‘repeated malarial infection lasting over a certain time. 1
THE SEASONAL PREVALENCE
In Tables Xa, Xb the distribution of 226 European cases of
blackwater fever in Northern Nigeria (1912) month by month is
given. The figures have not been corrected for monthly variations in
the European population as no figures are available for this purpose.
From 1900-1911 the total European population has increased from
about 175 to 775, but there is no reason to suppose that the increase
has not been a uniform one, spread over the whole of each year,
or that if in any year any disproportionate increase occurred in
any particular month it would not be equalized in other months in
other years.
Tabi.i Xa. —N. Nigeria. Showing the distribution by month of blackwater cases from
1899-1911.
489
Second Year
Month... ... ... ... ...j
13
1 1
]
| 1
16 '
1
l 1
l 7 I
18
19
20
21
22
23
24
!
Resident! .
1
42
1 1
t +* j 37
35
33
!
1
25
1
21
*7
12
10
9
5
Malaria cases .
■
1 21
29 18
1
9 !
*3
12
7
9
1
3
1
4
B.W. cases .
1
1
4 I 2
0 (
1
I
0
1
0
0
0
2
Table VIII. B. (compiled from previous table.)
Time of Residence.
First 6 months
First year
Second year
Number of residents .
69-5
61 -1
24
Cases of blackwater .
3
22
12
B.W. rate per ioo residents .
4*32
36-0
50*0
Of 100 cases of B.W. there occur .
5*02
41*86
58-14
i.e., Among a population of too 4*32 cases will occur in 1st 6 months residents.
36-0 „ „ ist year „
5°*° . „ 2nd year „
And of 100 cases of B.W. occurring during the 2 years’ period :
5*02 will occur in the ist 6 months.
41*86 „ „ ist year.
58*14 ,, „ 2nd year.
Malaria cases .
264
466
127
B.W. cases.
3
22
12
B.W. rate per ioo malaria cases ...
1*14
4*72
9*45
i.e.. For 100 cases of malaria in ist 6 months’ residents there will be 1*14 blackwater.
„ „ „ ist year’s „ „ 472 „
11 t n 2nd .,, ,, ,, 9*45 »
And of 100 cases of B.W. in 2 years there will be 8*05 among x malarial cases in the ist 6 months.
33*3 1 st » » *»t year.
66 69 „ „ n 2nd year.
The next table gives the distribution of 1,050 cases of which I
have found records. The data for the first six months and first year,
and possibly second year, are probably fairly correct, as they have
been taken from large and probably approximately equal popula¬
tions. But in the third and subsequent years we do not know what
proportion the population bears to that of the first year, and as I
have already pointed out, this is essential before a comparison can
be made. I should note, however, that Daniels in his figures has
made this correction, and that the difference from the uncorrected
figures was only slight.
494
The next Tables XlA, XIb show similar figures for Southern
Nigeria, Graham (r9i2).
Tabli XI a. —Southern Nigeria. Showing the distribution by month of blackwater cases from
1899-1911.
Table IX.—Blackwater Fever: Effect of Residence— continued.
49 i
492
I believe that we may deduce from these figures the fact
that blackwater is not a disease of the first six months or first year,
but mainly of the second year. I believe this fact is capable of
a simple explanation, viz., that blackwater fever only shows itself
in the majority of cases when the patient has been subjected to
‘repeated malarial infection lasting over a certain time. 1
THE SEASONAL PREVALENCE
In Tables Xa, Xb the distribution of 226 European cases of
blackwater fever in Northern Nigeria (1912) month by month is
given. The figures have not been corrected for monthly variations in
the European population as no figures are available for this purpose.
From 1900-1911 the total European population has increased from
about 175 to 775, but there is no reason to suppose that the increase
has not been a uniform one, spread over the whole of each year,
or that if in any year any disproportionate increase occurred in
any particular month it would not be equalized in other months in
other years.
Tabi.i Xa. —N. Nigeria. Showing the distribution by month of blackwater cases from
1899*1911.
Table Xb. —Northern Nigeria
Months
Actuals
Corrected for a
month of 30*4368
days
Departure 4- or —
from average 18 *8
1
30
29*5
4- 107
2
*4
•S-*
- 3*6
3
*4
*37
- 5 ’*
4
9
9 *i
- 97
5
8
7*9
— 10*9
6
9
9 *i
- 97
7
18
*77
— i*i
8
*7
265
+ 77
9
26
26*4
4 - 7*6
10
23
22*6
4 - 3*8
11
20
20*3
+ *'5
12
28
27*5
+ 87
1 226
I
225*5
Column i represents the months. Column 2 the actuals, taken
from the previous table. Column 3 the corrected figures for a
month of 30*4368 days. Column 4 the departure 4 - or — from the
average figure i8‘8. We observe in this table that the cases below
the average come together, and Mr. Stott has calculated for me
that it is 131 to 1 against the distribution being simply a random
one. Further, he has calculated that if we take into account the
amount of departure, that it is 1,000 to 1 in favour of the effect
being due to season.
The next table represents similar figures for the European
cases of blackwater admitted into Ancon Hospital.
Table XII.—Ancon Hospital, Panama. Showing the distribution by month of blackwater
cases among Europeans from 1908-1912.
icpS
« 9°9
1910
iqi I
1912
Totals
Departure
4 - or —
from the
average 20
January .
>7
1
5
2
28
- 8
February
1
9
4
6
3
2 3
+ 3
March .
3
10
3
4
3
23
+ 3
April .
1
6
3
8
1
19
— 1
May .
0
7
3
4
1
*5
~ 5
June .
0
3
4
7
1
15
- 5
J^y .
1
0
3
4
5
13
- 7
August .
1
4
3
8
4
20
0
September
0
1
2
10
6
19
October.
8
3
4
5
3
2 3
+ 3
November
11
7
3
6
2
• 29
+ 9
December
5
1
7
0
1
14
- 6
241
In Table XIII, I have given the uncorrected totals of Table XII
and the totals corrected for monthly variations in the European
population to show that the general conclusion is not materially
affected.
+97
Table XIII.—-Europeans at Ancon Hospital.
Average
Uncorrected
Corrected for population and for
LENGTH OF MONTHS
European
Population,
1908-1912
Blackwater
cases
admitted
1908-1912
Deviations
from
average
Blackwater
rate
per 10,000
Calculated
to make 241
Deviations
from
average
I .
6,061
28
+ 8
46-18
27-8
+ 77
2 .
6,112
23
+ 3
37-63
25-1
+ 5-0
3 .
6,090
23
+ 3
37*77
22-8
4- 27
4 .
6 , 43 6
! 9
— 1
29-52
18-4
- i *7
5 .
6,324
15
“ 5
23-72
! 4*3
- 5-8
6 .
6,143
15
- 5
24-42
15-2
- 4*9
7 .
6,138
*3
“ 7
21*18
12-8
- 7*3
8 .
5,982
20
0
33*43
20*1
— 00
9 .
5-953
T 9
— 1
31-92
19-8
- 0-3
10 .
6,241
23
+ 3
36*85
22-2
4 - 21
11 .
6,071
29
+ 9
47*77
297
4 - 9*6
12 .
6,290
14
- 6
22-26
13*4
- 67
73 , 4 <
241
392-65
241*6
Average ...
6,153
20
20-1
Accordingly, in Panama also, as Mr. Stott observes, we have
six cases below the average coming together, making a probability
of 131 to 1 in favour of a seasonal incidence, although in this case
our confidence is reduced by one-third owing to the average being
only over 5 years, instead of 13.
A similar conclusion can also be drawn from the figures for
blackwater for all employees admitted to the Commission hospitals
(Table XIV) showing that the Ancon figures are a fair sample of the
conditions existing over the whole zone.
For comparative purposes the monthly distribution of malaria is
also given, in which the seasonal malarial rise is clearly shown.
It should be noted that there are slight discrepancies between the
returns (Table XII) from Ancon Hospital (Europeans), privately
communicated by Dr. James, and those given in the monthly
official reports. I believe that the Ancon figures are reliable, as
they are the hospital records, whereas it is easily intelligible that
errors may occur in the official figures for the whole zone.
496
The next table represents similar figures for the European
cases of blackwater admitted into Ancon Hospital.
Table XII.—Ancon Hospital, Panama. Showing the distribution by month of blackwater
cases among Europeans from 1908-1912.
UPS
1909
1910
1911
1912
Totals
Departure
+ or -
from the
average 2c
January .
3
*7
1
5
2
28
- 8
February
1
9
4
6
3
23
- 3
March .
3
10
3
4
3
23
+ 3
April .
1
6
3
8
1
*9
- 1
May .
0
7
3
4
>
*5
- 5
June .
0
3
4
7
1
*5
~ 5
July .
1
0
3
4
5
13
- 7
August .
1
4
3
8
4
20
0
September
0
1
2
10
6
19
- 1
October.
8
3
4
5
3
23
+ 3
November
11
7
3
6
2
29
+ 9
December
5
1
7
0
1
*4
- 6
l
1
241
In Table XIII, I have given the uncorrected totals of Table XII
and the totals corrected for monthly variations in the European
population to show that the general conclusion is not materially
affected.
+97
Table XIII.—Europeans at Ancon Hospital,
Accordingly, in Panama also, as Mr. Stott observes, we have
six cases below the average coming together, making a probability
of 131 to 1 in favour of a seasonal incidence, although in this case
our confidence is reduced by one-third owing to the average being
only over 5 years, instead of 13.
A similar conclusion can also be drawn from the figures for
blackwater for all employees admitted to the Commission hospitals
(Table XIV) showing that the Ancon figures are a fair sample of the
conditions existing over the whole zone.
For comparative purposes the monthly distribution of malaria is
also given, in which the seasonal malarial rise is clearly shown.
It should be noted that there are slight discrepancies between the
returns (Table XII) from Ancon Hospital (Europeans), privately
communicated by Dr. James, and those given in the monthly
official reports. I believe that the Ancon figures are reliable, as
they are the hospital records, whereas it is easily intelligible that
errors may occur in the official figures for the whole zone.
500
Most of the statistics of blackwater and malaria do not extend
over a sufficiently long period to enable one to say that the correla¬
tion between the two is sufficiently close to make it fairly certain that
these are connected, or both subject to the same influence. This
applies, for example, to the figures for German East Africa,
Schilling (1910). An examination of these figures, kindly made
for me by Mr. Stott, Hon. Statistician to the School, certainly
suggests a correlation, but the period of years over which they
extend is too small to give a sufficiently satisfactory probability.
The next table—Table XVI—shows the amount of malaria and
blackwater in Panama in blacks and whites from 1906 to 1912. All
the table shows is that there has been a steady fall in malaria, and
also a fall in blackwater, but one cannot argue that there is
necessarily any relationship between the two, as the period is too
short. Ten years hence these figures with the additional observa¬
tions should be valuable. A probable fallacy is that we are not
dealing with a similarly constituted population each year, owing to
large emigration and immigration of labourers. If, for instance,
period of residence is a factor determining blackwater, then if the
number of people of each period of one, two, three, etc., years varies
each year, the blackwater rate may vary quite independently of the
malaria rate.
Table XVI.— Malaria and Blackwater Fever in the Panama Canal Zone.
i
1
i
Employees*
Annual Average
Malarial Admission
Rate
PER 1,000 PER ANNUM
Blackwater Admission
Rate
PER 1,000 PER ANNUM
Total
White |
Black
Total
White
Black j
Total
White 1
Black
1906
2 •’>547
5,400 ■
21,147
821*0
940*7
7877
i*88?
1
—
1907
39*238
10,709
28,429
4*8-7
753‘ 6
293*9
°*97 |
-
1908
43,890
* 2,383
3'>5°7
O
6
oo
r*
507-8
190*4
1*89
i 5'°9 |
0*63
1909
47>'&7
11,662
35o°5
211*4
366*2
*63-3
2*08
1 !
6*52
0-62
1910
50,802
13,021 l
37*781
*85-9
3727
121*5
1*04
3*07
| o*34
1911
48,876
12,251 |
36,625
181 *8
334*1
OC
6
2*07
1 6*6i
l
0*55
19] 2
5°, *93
12 '553 j
38)34°
op
O'
O
216*7
74-8
o*61
1*99
016
5oi
But a serious drawback to this table is that the statistics
regarding Americans and Europeans are massed together. The
malaria and blackwater fever in these two populations, one pro¬
tected, the other unprotected, or at least that does not protect itself,
are entirely different, as we shall now see.
In the following figures, however, we are able to separate the
data of the different races, and also we are able to do this for each
month over a period of five years. The data are based on the
admissions into Ancon Hospital, as in the monthly statistics for the
Canal Zone no separation into races is made. The difference
between the three races is briefly as follows :
(1) Americans—‘gold employees*; intelligent; living in well-
kept mosquito-proof houses; use of quinine at onset of fever
universal; receive pay when in hospital.
(2) Europeans—Spaniards, Italians and West Indian negroes ( !).
Those who desire it, live in mosquito-proof houses. Neglect
individual prophylaxis; indifferent to personal hygiene; receive no
pay when in hospital.
(3) Negroes—large majority live in cheap lodging houses, or in
huts; personal hygiene entirely lacking.
Now when we consider malaria and blackwater fever in these
races, as represented in Chart B, we see brought out very clearly
(1) that the incidence of malaria is mainly on the Europeans, much
less in the Americans and still less in the Negroes. The seasonal
incidence of malaria is shown very clearly in the case of Europeans
and Negroes, especially in the latter years. In Americans there
appears to be no clear seasonal incidence (of admissions), due no
doubt to the fact that ‘ they universally take quinine at the onset of
fever.’ (2) It is precisely in the Europeans, who suffer severely
from malaria, that we find blackwater, the cases being extremely
few in Americans and Negroes.
Now it appears to me that the explanation is obvious, viz., that
blackwater depends upon malaria. Of course it is possible to argue
that the relationship depends upon the fact that we are dealing
with two diseases both inoculated by the mosquito, or that those
suffering from malaria are debilitated and so open to the attack
of this hypothetical other disease, and even if for argument’s sake
we were to admit such hypotheses we should still be in the position
500
Most of the statistics of blackwater and malaria do not extend
over a sufficiently long period to enable one to say that the correla¬
tion between the two is sufficiently close to make it fairly certain that
these are connected, or both subject to the same influence. This
applies, for example, to the figures for German East Africa,
Schilling (1910). An examination of these figures, kindly made
for me by Mr. Stott, Hon. Statistician to the School, certainly
suggests a correlation, but the period of years over which they
extend is too small to give a sufficiently satisfactory probability.
The next table—Table XVI—shows the amount of malaria and
blackwater in Panama in blacks and whites from 1906 to 1912. All
the table shows is that there has been a steady fall in malaria, and
also a fall in blackwater, but one cannot argue that there is
necessarily any relationship between the two, as the period is too
short. Ten years hence these figures with the additional observa¬
tions should be valuable. A probable fallacy is that we are not
dealing with a similarly constituted population each year, owing to
large emigration and immigration of labourers. If, for instance,
period of residence is a factor determining blackwater, then if the
number of people of each period of one, two, three, etc., years varies
each year, the blackwater rate may vary quite independently of the
malaria rate.
Table XVI.—Malaria and Blackwater Fever in the Panama Canal Zone.
Employees’
Annual Average
Malarial Admission
Rate
per 1,000 PER ANNUM
Blackwater Admission
Rate
PER 1,000 PER ANNUM
Total
White |
Black
Total
White
Black
Total
White
Black
1906
*6,547
i
5,400
21.147
821*0
940-7
7 8 7'7
i-88 ?
—
1907
39,*3 8
!
10,709
28,4*9
4187
753-6
293*9
0-97
-
1908
43^90
■*,3 8 3 I
3*7507
280-0
507-8
190-4
1-89
5-09
0-63
1909
47: '67
1
11,662
35>5°5
2II-4
366-2
>63-3
2*08
6-52
0-62
1910
50,802
13,021
37>7 8 '
185-9
3727
121-5
1-04
3-07 |
o-34
1911
48,876
12,251 j
36,625
181-8
334**
I30-8
2-07
6-61
o*55
1912
50,893
1*1553 I
38,340
109-8
2167
74 -8
o-6i
j *‘99
o-io
But a serious drawback to this table is that the statistics
regarding Americans and Europeans are massed together. The
malaria and blackwater fever in these two populations, one pro¬
tected, the other unprotected, or at least that does not protect itself,
are entirely different, as we shall now see.
In the following figures, however, we are able to separate the
data of the different races, and also we are able to do this for each
month over a period of five years. The data are based on the
admissions into Ancon Hospital, as in the monthly statistics for the
Canal Zone no separation into races is made. The difference
between the three races is briefly as follows:
(1) Americans—‘gold employees'; intelligent; living in well-
kept mosquito-proof houses; use of quinine at onset of fever
universal; receive pay when in hospital.
(2) Europeans—Spaniards, Italians and West Indian negroes ( !).
Those who desire it, live in mosquito-proof houses. Neglect
individual prophylaxis; indifferent to personal hygiene; receive no
pay when in hospital.
(3) Negroes—large majority live in cheap lodging houses, or in
huts; personal hygiene entirely lacking.
Now when we consider malaria and blackwater fever in these
races, as represented in Chart B, we see brought out very clearly
(1) that the incidence of malaria is mainly on the Europeans, much
less in the Americans and still less in the Negroes. The seasonal
incidence of malaria is shown very clearly in the case of Europeans
and Negroes, especially in the latter years. In Americans there
appears to be no clear seasonal incidence (of admissions), due no
doubt to the fact that ‘ they universally take quinine at the onset of
fever.' (2) It is precisely in the Europeans, who suffer severely
from malaria, that we find blackwater, the cases being extremely
few in Americans and Negroes.
Now it appears to me that the explanation is obvious, viz., that
blackwater depends upon malaria. Of course it is possible to argue
that the relationship depends upon the fact that we are dealing
with two diseases both inoculated by the mosquito, or that those
suffering from malaria are debilitated and so open to the attack
of this hypothetical other disease, and even if for argument’s sake
we were to admit such hypotheses we should still be in the position
502
that this disease affects those suffering from malaria. But we
consider that the obvious explanation is the true one. And again,
the figures lend no support to the view that there is a quinine
haemoglobinuria distinct from blackwater fever, because, if so, we
should expect it in the Americans—the quinine takers.
Examining next the European admissions into Ancon Hospital
for 5 years for malaria and blackwater fever, respectively, we get
the results shown in Table XVII. Statistically there is a small but
negative correlation. If now, however, we move the blackwater
figures back for 4 months (i.e., make the blackwater follow the
malaria 4 months later) the result seen in the third series of
figures is got, and now there is a strong positive correlation,
expressed by r = + *50 + 04 (r = 1 implies perfect correlation),
tending to show that the two diseases are connected, for not only
is there a correlation between the seasonal and secular variations
which might occur, and yet not necessarily imply any connection
between the two diseases at all, but there is also a coincidence
between the magnitude of the oscillations of the two diseases, which,
so far as the figures go, suggests that there is a real connection
between the two.
Table XVII. — Europeans, Ancon Hospital
(1) Malaria Cases
Month
1908
* 9°9
1910
191 I
1912
5 vears
Departure
+ or -
from average
1092
■
*23
578
96
,zz
122
1,041
“ 5 *
2
uzl
397
96
IO8
-7
940
- *52
3
*54
244
,2 9
I2 9
*°5
761
- 33 *
4
103
161
136
1+8
76
624
- 468
5
106
*53
l6l
206
9 *
7*7
- 375
6
*77
164
1 317
523
116
*,297
| + 205
7
392
213
499
44 *
305
1,850
+ 758
S
4*9
*52
' 43 *
2*3
240
*455
+ 363
9
344
* 5 *
298
205
109
1,107
+ .5
10
59 *
146
233
*56
76
1
1,202
j +110
11
54 *
146
168
*25
9 *
1,07*
1 ~ 21
12
496
*72
*54
* *9
*03 |
*,044
- 48
00
rn
2-677
1 2,718
2,495
1,561
13,109
S ®3
(2) Blackwattr Casks
1
Month
1908
; i
1909
1910
1911
1912
5 years
f Departure
4* or —
from average
20
1
3
*7
1
5
2
28
+ 8
2
1
9
4
6
3
23
+ 3
3
3
10
3
4
3
23
+ 3
4
1
6
3
8
1
19 1
— 1
5
-
7
3
4
1
15
“ 5
6
—
3
4
7
1
15
- 5
7
1
1 0
3
4
5
*3
- 7
8
1
1
1 4
;
3
8
4
20
0
9
—
' *
I
2
1 10
6
*9
— 1
10
8
1 3
4
i 5
3
23
+ 3
11
11
1 7
3
1
6
2
29
+ 9
12
5
I
7
0
1
14
- 6
34
68
40
67
:
3 *
241
(3) Blackwater Figures Moved Four Months Back
Month
00
■ 9°9
1910
1911
1912
5 years
Departure
4 - or -
from average
20
1
—
7
3
4
■
*5
- 5
2
—
3
4
7
-
>
*5
- 5
3
1
0
3
4
5
*3
- 7
4
1
4
3
8
4
20
0
■
5
—
1
2
10
6
*9
- *
6
8
3
4
5
3
23
+ 3
7
11
7
3
6
2
29
+ 9
8
5
1
7
0
1
H
- 6
9
17
*
5
2
1
26
4 - 6
10
9
4
6
3
2
24
+ 4
11
10
3
4
3
3
23
+ 3
12
6
3
8
1
2
20
0
68
37
52
53
3 *
241
SECOND ATTACKS
They occur in about 10%. I have not so far examined very
extensive records. A very interesting feature about these attacks
is that they are most common in the first year, and next so in the
first six months. These data are, however, subject to correction
for the population in each year period, but, I believe, are
approximately correct. Whether this feature indicates increased
susceptibility to attack, or whether they are for the most part really
relapses, further consideration must show.
Table XVIII.—Blackwater Fever : Second attack®.
6 months
1 st year
2nd year
i
3rd and
later years 1
Total
Graham 191 2) .
7
1 5
! 3
8
26
Christophers & Bentley J
(1908)
8
12
2
3
17
Decks Si James (19! 1} • ••'
8
>4
1
2
_
16
Totals ...
2 3
+l
7
11
59
Percentages ...
39
! 69-5
i
ii *9
1
18-6
I have to acknowledge the great kindness of Dr. James, Ancon
Hospital, Panama, in supplying me with many statistical data and
information on many points that arose during a consideration of the
Panama figures. I also am indebted to Colonel Gorgas for similar
information and interest in my work. To Mr. Stott I am indebted
for the great trouble he has taken in examining my figures from
the statistical point of view. Finally, I must acknowledge very
gratefully the laborious work of Mr. Drawz, Bibliographer to the
School, in compiling the data from the Panama Canal Commission
Reports and elsewhere.
* By a second attack I mean one that has occurred after the discharge of the patient
from hospital.
REFERENCES
Barratt, J. O. W., and Yorke, W. (1909). An investigation into the production of black-
water. Ann. Trop. Med. Parasitol., Liverpool, Oct. 1, III, 1, pp. 1-256. With charts,
figs, and plates.
Birenger-Feraud, L. J. B. (1874). De la fievre bilieuse melanurique des pays chauds comparee
avec la fievre jaune. Etude clinique faite au Seagal. Pp. 442, 8°., Paris.
Brault, J. (1903). Note sur la fievre hemoglobinurique en Alg^ric. Janus, Harlem, VIII,
pp. 561-566.
Brem, W. V. (1906). Malarial hemoglobinuria. Joum. Amer. Med. Assoc., Chicago, Dec. 8,
XLVII, pp. 1896-1904.
- (1911). Studies of malaria in Panama. II. Treatment of blackwater fever. Pernicious
malaria with hemoglobinuria and erythrolytic hemoglobinuria. Arch. Intern. Med.,
Chicago, Feb., VII, pp. 153-181.
Broden, A. (1906). L’Hemoglobinurie au Congo. Trav. Lab. Med. Leopoldville, Bruxelles,
II, pp. 1-70.
Burot, F., and Legrand, A. (1897). Les troupes coloniales. Paris. (Cited by Mense, Arch.
Schiffs. Trop. Hyg., 1899, p. 217.)
Campenhout and Dryepondt (1901). Fievre bilieuse hemoglobinurique. Trav. Lab. Med.
Leopoldville, Bruxelles, I (1899-1900), pp. 51-117.
Capogrossi, A. (1910). Un caso di emoglobinuria da terzana primaverile con albuminuria
secondaria. Atti Soc. Stud. Malaria, Rome, XI, pp. 703-708.
Cardamatis, J. P. (1902). La fiivre bilieuse hemoglobinurique observee en Gr^ce.—Statistique
—Etiologie—Traitment. Rev. Med. Afrique Nord, Paris, pp. 1-53. (Reprint.)
- (1911). Observations microbiologiques et histologiques sur 80 cas de fievre bilieuse hemo¬
globinurique. Centralbl. Bakt. (etc.), I Abt., Orig., Jena, LXI, pp. 378-381.
Carducci, A. (1907). Le emoglobinurie da chinino in individui affetti da terzana primaverile
e da quartana. Atti Soc. Stud. Malaria, Rome, VIII, pp. 225-242.
Costa, B. F. B. da (1906). Estudos sobre a etiologia da febre biliosa hemoglobinurica. Arch.
Hyg. Path. Exot., Lisbon, I, 2, pp. 218-273.
Christophers, S. R., and Bentley, C. A. (1908). Blackwater fever. Sc. Mem. Off. Med.
Dep. Gov. India, Simla, new ser., No. 35, Pp. 239.
Daniels, C. W. (1901) Notes on ‘ Blackwater Fever * in British Central Africa. Rep. Malaria
Comm. Roy. Soc., London, 5th ser., pp. 44-78. With charts.
Deaderick, W. H. (1907). A preliminary report of calcium chloride in the treatment of
haemoglobinuric fever. Joum. Trop. Med. Hyg., London, Dec. 16, X, 24, pp. 393-394.
- (1907-08). Hemoglobinuric fever. Memphis Med. Monthly, Dec.-March. (Reprint.)
Deeks, W. E., and James, W. M. (1911). A report on hemoglobinuric fever in the Canal
Zone. A study of its etiology and treatment. Pp. 177. With charts and tables.
Mount Hope, C.Z.
Frere, J. E. (1910). Two cases of blackwater fever. Lancet, Lond., June 18,1, pp. 1716-1717.
Gastou, P., and Dufougere, W. (1911). Paludisme et fievre bilieuse hemoglobinurique.
Bull. Soc. Path. Exot., Paris, May 10, IV, 5, pp. 301-303.
German East Africa (1909-10). Med. Ber. Deutsch. Schutzgeb., 1909-10, Berlin, pp. 113-114.
Graham, W. M. (1912). Report on blackwater fever in Southern Nigeria, 1899-1911. Pp. 72.
With plates, charts, and 1 map. London.
504
SECOND ATTACKS*
They occur in about 10%. I have not so far examined very
extensive records. A very interesting feature about these attacks
is that they are most common in the first year, and next so in the
first six months. These data are, however, subject to correction
for the population in each year period, but, I believe, are
approximately correct. Whether this feature indicates increased
susceptibility to attack, or whether they are for the most part really
relapses, further consideration must show.
Table XVIII.—Blackwater Fever : Second attacks.
i 1
1 6 months j
1st year
| 2nd year
3rd and
later years
1 Total
Graham (191 1)
•■! 7
1 5
3
8
26
Christophers & Bentley
(* 9 °*)
8
12
2
3
*7
Deeks & James (1911)
••! 8
14
2
—
1 16
Totals ...
2 3
4 i
7
11
59
Percentages .
39
69-5
1 1’9
!
18*6 j
I have to acknowledge the great kindness of Dr. James, Ancon
Hospital, Panama, in supplying me with many statistical data and
information on many points that arose during a consideration of the
Panama figures. I also am indebted to Colonel Gorgas for similar
information and interest in my work. To Mr. Stott I am indebted
for the great trouble he has taken in examining my figures from
the statistical point of view. Finally, I must acknowledge very
gratefully the laborious work of Mr. Drawz, Bibliographer to the
School, in compiling the data from the Panama Canal Commission
Reports and elsewhere.
* By a second attack I mean one that has occurred after the discharge of the patient
from hospital.
REFERENCES
Barratt, J. O. W., and Yorke, W. (1909). An investigation into the production of black-
water. Ann. Trop. Med. Parasitol., Liverpool, Oct. 1, III, 1, pp. 1-256. With charts,
figs, and plates.
Berenger-Feraud, L. J. B. (1874). De la fievre bilieuse m&anurique des pays chauds comparee
avec la ftevre jaune. Etude clinique faite au S6n6gal. Pp. 442, 8°., Paris.
Brault, J. (1903). Note sur la fievre h£moglobinurique en Alg6ric. Janus, Harlem, VIII,
pp. 561-566.
Brem, W. V. (1906). Malarial hemoglobinuria. Journ. Amer. Med. Assoc., Chicago, Dec. 8,
XLVII, pp. 1896-1904.
- (1911). Studies of malaria in Panama. II. Treatment of blackwater fever. Pernicious
malaria with hemoglobinuria and erythrolytic hemoglobinuria. Arch. Intern. Med.,
Chicago, Feb., VII, pp. 153-181.
Broden, A. (1906). L’Hemoglobinurie au Congo. Trav. Lab. Med. Leopoldville, Bruxelles,
II, pp. 1-70.
Burot, F., and Legrand, A. (1897). Les troupes coloniales. Paris. (Cited by Mense, Arch.
Schiffs. Trop. Hyg., 1899, p. 217.)
Campenhout and Dryepondt (1901). Fidvre bilieuse hcmoglobinurique. Trav. Lab. Med.
Leopoldville, Bruxelles, I (1899-1900), pp. 51-117.
Capogrossi, A. (1910). Un caso di emoglobinuria da terzana primaverile con albuminuria
secondaria. Atti Soc. Stud. Malaria, Rome, XI, pp. 703-708.
Card amahs, J. P. (1902). La fievre bilieuse hcmoglobinurique observee en GrCce.—Statistique
—Etiologie—Traitment. Rev. MCd. Afrique Nord, Paris, pp. 1-53. (Reprint.)
- (1911). Observations microbiologiques et histologiques sur 80 cas de fievre bilieuse hCmo-
globinurique. Centralbl. Bakt. (etc.), I Abt., Orig., Jena, LXI, pp. 378-381.
Carducci, A. (1907). Le emoglobinurie da chinino in individui affetti da terzana primaverile
e da quartana. Atti Soc. Stud. Malaria, Rome, VIII, pp. 225-242.
Costa, B. F. B. da (1906). Estudos sobre a etiologia da febre biliosa hemoglobinurica. Arch.
Hyg. Path. Exot., Lisbon, I, 2, pp. 218-273.
Christophers, S. R., and Bentley, C. A. (1908). Blackwater fever. Sc. Mem. Off. Med.
Dep. Gov. India, Simla, new ser., No. 35, Pp. 239.
Daniels, C. W. (1901) Notes on 4 Blackwater Fever * in British Central Africa. Rep. Malaria
Comm. Roy. Soc., London, 5th ser., pp. 44-78. With charts.
Deaderick, W. H. (1907). A preliminary report of calcium chloride in the treatment of
haemoglobinuric fever. Joum. Trop. Med. Hyg., London, Dec. 16, X, 24, pp. 393-394.
- (1907-08). Hemoglobinurie fever. Memphis Med. Monthly, Dec.-March. (Reprint.)
Deeks, W. E., and James, W. M. (1911). A report on hemoglobinuric fever in the Canal
Zone. A study of its etiology and treatment. Pp. 177. With charts and tables.
Mount Hope, C.Z.
Frere, J. E. (1910). Two cases of blackwater fever. Lancet, Lond., June 18,1, pp. 1716-1717.
Gastou, P., and Dupougere, W. (1911). Paludisme et fiCvre bilieuse htmoglobinurique.
Bull. Soc. Path. Exot., Paris, May 10, IV, 5, pp. 301-303.
German East Africa (1909-10). Med. Ber. Deutsch. Schutzgeb., 1909-10, Berlin, pp. 113-114.
Graham, W. M. (1912). Report on blackwater fever in Southern Nigeria, 1899-1911. Pp. 72.
With plates, charts, and 1 map. London.
Grattan, W. H. (1907). A note on blackwater fever in Sierra Leone. Journ. R.A.M.C.,
London, Sept., IX, 3, pp. 237-243.
Howard, R. (1907). A cate of blackwater fever occurring after twenty-three year*’ residence
in Central Africa. Journ. Trop. Med. Hyg., London, March 1, X, 5, p. 81.
Jung els, — (1911). Vorliufige Mitteilungen fiber mehrere Fille von Schwarzwasserfieber,
beobachtet bei ostafrikanischen Negern. Arch. Schiffs. Trop. Hyg., Leipzig, XV, 11,
pp. 361-362. (Notizen a.d. Tropenprazis, No. 2.)
K.LXINI, F. K. (1901). Uber Schwarzwasserfieber. Zeitschr. Hyg. Inf., Leipzig, XXXVIII,
pp. 472-486. With charts.
Koch, R. (1899). Ueber Schwarzwasserfieber (Hamoglobinurie). Zeitschr. Hyg. Inf.,
Leipzig, XXX, pp. 295-327.
Kravss, W. (1904). Malarial hemoglobinuria. Intemat. Clin., Phila., 14th ser., II, pp. 52-67,
With charts and plates.
Kulz, L. (1908). Uber einen Fall von Nephrotomie bei Anurie nach Schwarzwasserfieber.
Arch. Schiffs. Trop. Hyg., Leipzig, XII, 15, pp. 508-510. (Notizen a.d. Tropenpraxii,
No. 5.)
- (1910). Be it rag zu einer Cholestearin-Therapie des Sch warz waste rfiebers. Arch.
Schiffs. Trop. Hyg., Leipzig, XIV, 23, pp. 739-743*
Lovelace, C. (1913). The etiology and treatment of hemoglobinuric fever.—Report of five
hundred and fourteen cases. Arch. Intern. Med., Chicago, June 15, XI, 6, pp. 674-684.
McElroy, — (>905)* Memphis Med. Month., May-Junc. (Cited by Deaderick, 1907-08,
P . 11.)
Marshall, D. G. (1910). A case of blackwater fever. Lancet, London, May 14, 1 , pp. 1333-
1334 *
Master man, E. W. G. (1906). Haemoglobinuric fever in Syria. Brit. Med. Journ., London,
Feb. 10, I, pp. 314-315. With chart.
Matta, A. A. da (1912). A febre biliosa hemoglobinurica no Amazonas e o seu tratamento
pela Cecropia. Rev. Med. S. Paulo, S. Paulo, Sept. 30, XV, 18, pp. 357-364. With
4 charts.
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fever. Journ. Lond. Sch. Trop. Med., London, July, I, 3, pp. 281-283.
Orme, B. (1908). Cases of blackwater fever in the Malay Peninsula. Journ. Trop. Med. Hyg.,
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5°9
THE GROWTH AND SPORULATION
OF THE BENIGN AND MALIGNANT
TERTIAN MALARIAL PARASITES
IN THE CULTURE TUBE AND IN
THE HUMAN HOST
BY
JOHN GORDON THOMSON, M.A., M.B., Ch.B. (Edin.)
SIR EDWIN DURNING-LAWRENCE RESEARCH ASSISTANT, AND
PATHOLOGIST, ROYAL SOUTHERN HOSPITAL
AND
DAVID THOMSON, M.B., Ch.B. (Edin.), D.P.H. (Cantab.)
GROCERS' RESEARCH SCHOLAR, LATELY CLINICAL AND PATHOLOGICAL
RESEARCH ASSISTANT, SCHOOL OF TROPICAL MEDICINE, LIVERPOOL
(.Received for publication 14 September, 1913)
Plates XXXIV, XXXV
CONTENTS
Prefatory note by Sir Ronald Ross .
Introduction .
Cultivation Technique.
Further observations on cultivation.
Optimum temperature, Haemolysis, Clumping .
Resistant forms.
Morphology of P. falciparum in the culture tube .
„ ,, „ „ human host .
,, ,, P. vivax in the culture tube
„ „ „ „ human host
Summary.
References.
Explanation of Plates .
page
509
510
510
5 **
5*2
5*3
5*3
516
518
5*9
520
52*
522
PREFATORY NOTE BY SIR RONALD ROSS
Researches on the cultivation of the parasites of malaria in
Liverpool were commenced some time ago at my suggestion by
Dr. Sint on, and then, with better success, by Drs. J. G. Thomson
and McLellan, and by Dr. D. Thomson. We are greatly obliged
to Sir Edwin Durning-Lawrence, Bart., for giving us the services
of Dr. J. G. Thomson for this important enquiry.
2ir/ May, 1913.
Ronald Ross.
5io
INTRODUCTION
The successful cultivation of malarial parasites was first
announced by Bass and Johns (1912). Since then several workers,
Thomson and McLellan (1912), J. G. Thomson and D. Thomson
(1913) and Ziemann (1913) have successfully repeated these
cultivation experiments. This achievement has led the way to new
discoveries regarding the malarial parasite, and suggests that it
may be possible to cultivate in vitro any protozoal parasite
however specialised it may be.
So far, only the asexual generation of the malarial parasite has
been grown in vitro . The next step is to cultivate the sexual
generation as it occurs in the human host and in the mosquito.
We have attempted both, but so far without definite success. We
have no doubt, however, that this will also be accomplished sooner
or later. In 1911, J. G. Thomson and Sinton successfully
cultivated the human trypanosome, and the development they
obtained was apparently that which takes place in the stomach of
the tsetse fly. Joukoff (1913) states that he has cultivated the
mosquito cycle of the malarial parasite, though his results have not
yet been confirmed. There is no reason to doubt that the phases
of development of protozoa in insects may be produced in the
culture tube.
An abridged account of these researches, illustrated by Plate
XXXIV, was published in the Proceedings of the Royal Society,
B, Vol. 87 (1913), pp. 77-87. We have much pleasure in
returning thanks to the Royal Society for permission to reprint
Plate XXXIV.
We wish to thank Dr. H. B. Fantham for his very valuable
help during our work on this subject.
CULTIVATION TECHNIQUE
Our method of cultivation is almost the same as that of
Bass and Johns, except that it is less complicated. Ten c.c. of
blood is drawn from a vein and transferred to a sterile test tube
containing a thick wire leading to the bottom of the tube from
the cotton wool plug. 1 /10 c.c. of a 50 per cent, aqueous solution of
glucose is added to this tube, preferably before adding the blood.
The blood is defibrinated by gently stirring with the thick wire.
Defibrination should be complete in about 5 minutes. The wire
with the clot is then removed and the blood is poured into several
smaller sterile tubes (about one inch column of blood in each).
A rubber cap is placed over the cotton plugs to prevent evaporation
and the tubes are then transferred (standing upright) to an
incubator at a temperature of 37 0 to 4i°C. The corpuscles settle
in a short time, leaving about one half-inch of clear serum at the
top. It is apparently unnecessary to remove the leucocytes by
centrifugalisation.
FURTHER OBSERVATIONS ON CULTIVATION
We have grown four complete generations of parasites in one
tube by the above method, and we do not see why their growth
should not continue indefinitely, provided fresh serum and
corpuscles be added. It is not the presence of leucocytes which
prevents further development. This is due to degenerative changes
taking place in the corpuscles and serum. If the serum and
corpuscles be kept in a sterile condition in an ice chest they remain
unchanged for a long time; but at the temperature required for the
growth of the parasites visible changes take place in a few days.
The corpuscles become fragmented and form a brownish d6bris, and
the serum becomes dark brown in colour. When this occurs the
parasites are unable to continue their developmental cycle. Bass and
Johns in their original paper (1912) stated that the parasites grow
only on the surface layer of corpuscles, and that no growth took
place in the deeper layers. They also stated that the serum
destroyed the parasites when they escaped from the corpuscles, so
that when the spores escape they must enter immediately into a
contiguous corpuscle in order to survive. The blood also, in their
opinion, required to be heated to a temperature of 40° C. to destroy
the complement in the serum; furthermore, in order to cultivate
several generations, the leucocytes had to be removed. We do not
believe that they have sufficient evidence for these deductions. We
find that the parasites develop even in the very deepest layer of the
column of corpuscles. Also, it seems rather a contradiction that
the parasites should grow best on the surface layer of corpuscles
5io
INTRODUCTION
The successful cultivation of malarial parasites was first
announced by Bass and Johns (1912). Since then several workers,
Thomson and McLellan (1912), J. G. Thomson and D. Thomson
(1913) and Ziemann (1913) have successfully repeated these
cultivation experiments. This achievement has led the way to new
discoveries regarding the malarial parasite, and suggests that it
may be possible to cultivate in vitro any protozoal parasite
however specialised it may be.
So far, only the asexual generation of the malarial parasite has
been grown in vitro . The next step is to cultivate the sexual
generation as it occurs in the human host and in the mosquito.
We have attempted both, but so far without definite success. We
have no doubt, however, that this will also be accomplished sooner
or later. In 1911, J. G. Thomson and Sinton successfully
cultivated the human trypanosome, and the development they
obtained was apparently that which takes place in the stomach of
the tsetse fly. Joukoff (1913) states that he has cultivated the
mosquito cycle of the malarial parasite, though his results have not
yet been confirmed. There is no reason to doubt that the phases
of development of protozoa in insects may be produced in the
culture tube.
An abridged account of these researches, illustrated by Plate
XXXIV, was published in the Proceedings of the Royal Society,
B, Vol. 87 (1913), pp. 77-87. We have much pleasure in
returning thanks to the Royal Society for permission to reprint
Plate XXXIV.
We wish to thank Dr. H. B. Fantham for his very valuable
help during our work on this subject.
CULTIVATION TECHNIQUE
Our method of cultivation is almost the same as that of
Bass and Johns, except that it is less complicated. Ten c.c. of
blood is drawn from a vein and transferred to a sterile test tube
containing a thick wire leading to the bottom of the tube from
the cotton wool plug. 1 /10 c.c. of a 50 per cent, aqueous solution of
glucose is added to this tube, preferably before adding the blood.
The blood is defibrinated by gently stirring with the thick wire.
Defibrination should be complete in about 5 minutes. The wire
with the clot is then removed and the blood is poured into several
smaller sterile tubes (about one inch column of blood in each).
A rubber cap is placed over the cotton plugs to prevent evaporation
and the tubes are then transferred (standing upright) to an
incubator at a temperature of 37 0 to 4i°C. The corpuscles settle
in a short time, leaving about one half-inch of clear serum at the
top. It is apparently unnecessary to remove the leucocytes by
centrifugalisation.
FURTHER OBSERVATIONS ON CULTIVATION
We have grown four complete generations of parasites in one
tube by the above method, and we do not see why their growth
should not continue indefinitely, provided fresh serum and
corpuscles be added. It is not the presence of leucocytes which
prevents further development. This is due to degenerative changes
taking place in the corpuscles and serum. If the serum and
corpuscles be kept in a sterile condition in an ice chest they remain
unchanged for a long time; but at the temperature required for the
growth of the parasites visible changes take place in a few days.
The corpuscles become fragmented and form a brownish debris, and
the serum becomes dark brown in colour. When this occurs the
parasites are unable to continue their developmental cycle. Bass and
Johns in their original paper (1912) stated that the parasites grow
only on the surface layer of corpuscles, and that no growth took
place in the deeper layers. They also stated that the serum
destroyed the parasites when they escaped from the corpuscles, so
that when the spores escape they must enter immediately into a
contiguous corpuscle in order to survive. The blood also, in their
opinion, required to be heated to a temperature of 40° C. to destroy
the complement in the serum; furthermore, in order to cultivate
several generations, the leucocytes had to be removed. We do not
believe that they have sufficient evidence for these deductions. We
find that the parasites develop even in the very deepest layer of the
column of corpuscles. Also, it seems rather a contradiction that
the parasites should grow best on the surface layer of corpuscles
512
next to the serum and the leucocytes which are supposed to destroy
them. No doubt the leucocytes ingest some of the parasites, but
they are never able to ingest all of them. Again, in one of our
most successful cultures the temperature of the blood never exceeded
38° C., and we have found that they are able to grow at a tem¬
perature as low as 36° C. This is rather against the theory of
complement destruction. In an able paper by Mary Rowley Lawson
(1913), considerable evidence is brought forward to show that the
parasites are extra-corpuscular during their entire development.
If this is true, then the parasites would be constantly in contact with
the serum which is supposed to destroy them. It appears to us
that the only conditions necessary for the successful cultivation of
the parasites are fresh corpuscles, fresh serum, a temperature of
37 0 to 4i°C. and the requisite amount of glucose. The presence
of leucocytes and complement is apparently immaterial.
THE OPTIMUM TEMPERATURE
In our opinion the optimum temperature for cultivation is
about 38° C. On two occasions we incubated identical culture
tubes from the same patient, some at 37 0 C. and some at 41 0 C. On
both the occasions the parasites developed much more successfully
in the tubes incubated at 37°C.
HAEMOLYSIS IN THE CULTURE TUBES
This is a rare occurrence, having been seen only once out of
fifteen cases. A kind of haemolysis occurs after the blood has
remained in the incubator for several days. The corpuscles
degenerate into a brownish debris, and the serum shows a brown
discoloration.
CLUMPING OF THE MALIGNANT TERTIAN PARASITES
This phenomenon has been observed in all our malignant tertian
cultures (twelve cases). It occurs even when the parasites are
scarce. It is best observed in wet films, as smearing of the blood
tends to break up the clumps. No tendency to clumping occurred
in our benign tertian cultures.
of the young merozoites escape ingestion by the leucocytes and enter
a new red cell. The same culture examined in three days again
showed segmenting forms (fig. 14); but in this case the spores were
never so numerous as 32, and this can easily be explained by the
adverse conditions which have now developed in the culture tube.
These conditions did not, however, prevent the young parasites
attempting to segment. In four and a half days (fig. 15) we again
only found young rings, which represent the beginning of a third
generation; and in six days sporulating forms were again found
(fig. 16). Here, only eight spores have formed, and it was evident
that the power of the parasite to undergo full segmentation was
getting gradually less. On the seventh day, again, only very
young plasmodia were found, which represented a fourth genera¬
tion, and these again attempted to sporulate (figs. 18 and 19) on
the eighth and ninth days respectively. On the tenth day only
young parasites again were found, which represented a fifth genera¬
tion. No further development, however, occurred, as the conditions
in the culture were gradually becoming more and more adverse.
Thus we have evidence that the parasites will, under very suitable
conditions, actually proceed in the original culture tube through
four complete generations without the medium being in any way
renewed.
We now wish to draw particular attention to several points of
great interest which occur in the cultures of P. falciparum . In
these there is a definite tendency for the parasites to clump together
into masses immediately the circular mass of pigment appears, and
even before segmentation begins. A mass of seven parasites is
seen in the microphotograph, Plate XXXV, fig. 2. This photograph
was taken after twelve hours* incubation at 38° C. In all of these note
the dark circular mass of pigment, the parasite having a diameter of
about 6 n. At this stage the chromatin is dispersed into the proto¬
plasm of the plasmodium, and as yet shows no division into daughter
cells. This tendency to clump becomes more marked when
sporulation actually begins, and fig. 3, Plate XXXV, shows a
large mass of sporulating parasites, the photograph being taken
after twenty-five hours* cultivation. This picture shows all stages
of sporulation and illustrates in a very striking manner the tendency
of the parasites to agglomerate into masses. This clumping in
to cultivate the malignant tertain parasite, and these have usually
been from patients to whom quinine had been administered.
On two occasions complete sporulation did not take place till
after fifty hours* incubation at 41 ° C. On one of these occasions the
parasites completed several generations, as illustrated by the accom¬
panying coloured plate (Plate XXXIV, figs. 1-20). We found that
maximum segmentation took place in fifty-two hours. Division
of the chromatin into daughter nuclei began in about thirty-six
hours (figs. 4 and 5). In forty-seven hours the number of spores
had increased (figs. 6 and 7), and more or less complete segmenta¬
tion took place in fifty-two hours. Fig. 11 shows a parasite which
has produced thirty-two daughter cells, and these have broken loose
from the corpuscle. It is to be noticed that in the peripheral blood
the small ring parasites show no pigment. After incubation the
rings gradually grow in size (fig. 3), and at a certain stage,
immediately previous to the splitting of the chromatin a round
compact mass of pigment appears, usually situated near one margin
of the parasite, which now measures about 5 fi or 6 fi in longest
diameter. In the culture under discussion this round mass of
pigment was seen in thirty-six hours (figs. 4 and 5). J. G. Thomson
and S. W. McLellan (1912) found that the pigment had collected
in twelve hours. The parasites immediately previous to segmenta¬
tion measure about 5 /* to 6/* in their longest diameter, and all show
the circular mass of compact pigment. We may call this stage the
pre-segmenting stage. The chromatin now begins to split into two,
and if segmentation is complete thirty-two daughter cells may be
formed (fig. 11). All stages of segmentation can be found, from
two spores up to thirty-two spores as a maximum. During
segmentation the circular mass of pigment takes a central position,
and the spores form in a circular arrangement around these (figs.
7 to 11). In this culture segmenting forms were found at different
stages up to fifty-six hours, this being, no doubt, due to the fact
that the parasites were not all of the same age when introduced
into the culture tubes.
In seventy-five hours all segmenting forms had disappeared and
only very young parasites were found, about 1*5/1 in diameter and
containing no vacuole (fig. 13). This represents the beginning of
a second generation. It is to be concluded, therefore, that many
of the young merozoites escape ingestion by the leucocytes and enter
a new red cell. The same culture examined in three days again
showed segmenting forms (fig. 14); but in this case the spores were
never so numerous as 32, and this can easily be explained by the
adverse conditions which have now developed in the culture tube.
These conditions did not, however, prevent the young parasites
attempting to segment. In four and a half days (fig. 15) we again
only found young rings, which represent the beginning of a third
generation; and in six days sporulating forms were again found
(fig. 16). Here, only eight spores have formed, and it was evident
that the power of the parasite to undergo full segmentation was
getting gradually less. On the seventh day, again, only very
young plasmodia were found, which represented a fourth genera¬
tion, and these again attempted to sporulate (figs. 18 and 19) on
the eighth and ninth days respectively. On the tenth day only
young parasites again were found, which represented a fifth genera¬
tion. No further development, however, occurred, as the conditions
in the culture were gradually becoming more and more adverse.
Thus we have evidence that the parasites will, under very suitable
conditions, actually proceed in the original culture tube through
four complete generations without the medium being in any way
renewed.
We now wish to draw particular attention to several points of
great interest which occur in the cultures of P. falciparum . In
these there is a definite tendency for the parasites to clump together
into masses immediately the circular mass of pigment appears, and
even before segmentation begins. A mass of seven parasites is
seen in the microphotograph, Plate XXXV, fig. 2. This photograph
was taken after twelve hours’ incubation at 38° C. In all of these note
the dark circular mass of pigment, the parasite having a diameter of
about 6 fi. At this stage the chromatin is dispersed into the proto¬
plasm of the plasmodium, and as yet shows no division into daughter
cells. This tendency to clump becomes more marked when
sporulation actually begins, and fig. 3, Plate XXXV, shows a
large mass of sporulating parasites, the photograph being taken
after twenty-five hours’ cultivation. This picture shows all stages
of sporulation and illustrates in a very striking manner the tendency
of the parasites to agglomerate into masses. This clumping in
of course, that 32 spores are always produced, even under favour¬
able conditions, without quinine administration. It is likely that
the numbers produced vary considerably, just as in the case of
benign tertian, which produces numbers varying from 16 to 26.
The clumping phenomenon in the human host is represented in Plate
XXXV, fig. 5. This clump was, as already stated, found by
Dr. Cropper in a peripheral blood smear. A similar phenomenon
is seen in the brain capillaries of cases of comatose malaria.
THE MORPHOLOGY OF PLASMODIUM VIVAX IN CULTURE
Here we are studying a parasite in which all stages, from small
rings up to full sporulation, may be seen in the peripheral blood.
Sporulation does not necessarily occur in the internal organs, and it
is quite usual to find these forms in ordinary peripheral blood
smears. When cultivating this parasite, therefore, it is important
to draw the blood from the patient when the young forms pre¬
dominate, so that it is then certain whether or not we are obtaining
further development in the culture tubes. In the culture illustrated
in Plate XXXIV, figs. 21 to 30, we obtained the blood when young
rings (fig. 21) predominated, and no segmenting forms were found.
After eight hours’ incubation at a temperature of 39°C., a marked
increase in size of the parasites was noted, figs. 22, 23 and 24, and
pigment was now evident, being scattered throughout the proto¬
plasm in fine granules. This arrangement of the pigment in
P. vivax is in marked contrast to what occurs in P. falciparum ,
where the pigment always becomes arranged in a dense circular
mass from the commencement. After 20 to 29 hours’ incubation
(figs. 25-30), sporulation was seen at different stages, the pigment
being collected into a loose mass of granules in the centre of the
parasite. Fig. 29 shows a parasite with 15 daughter cells, and
we have found on several occasions sixteen spores, which we think
is the usual maximum of daughter cells found in P. vivax , although
again we find all stages, from two spores up to sixteen, according
to the stage of development at which we examine the cultures.
We are quite certain that the spores are never so numerous as in the
malignant tertian parasite. In these cultures clumping has not been
found, and this explains why the parasites do not tend to be
5*7
rule, in the peripheral blood. This is due to the fact that when
these parasites have grown larger than the ring stage, the containing
corpuscles stick together, forming clumps which are unable to
circulate, and thus are arrested in the fine capillaries of the inner
organs. The study of the further stages of this parasite is
obtained by examining smears of the inner organs, such as the
spleen, liver, brain, etc., of deceased patients. The autopsy
smears of over one hundred cases have recently been examined by
one of us (D. T.) in conjunction with Dr. W. M. James, in
Panama (the results of this work will be published later). In such
smears the parasite is found in different stages of development.
In some, pre-segmenters only are found; some show only young
rings, and others only sporulating forms. This depends entirely
on the stage of development of the parasite at the time of the
patients* death. In some cases, of course, several stages of
development are found, since one patient may contain several
broods of parasites which are in different stages of development.
In only one case did we find the parasites in the stage of maximum
sporulation. All of them contained over 20 spores, and 13 per
cent, had 32 spores. One of these is represented in the microphoto¬
graph (Plate XXXV, fig. 4). We think that there can be little
doubt that we have proved conclusively, both from observations in
the culture tube and in the human host, that P. falciparum is
capable of producing a maximum of 32 spores under favourable
circumstances. This is a much larger number than has been pre¬
viously given in the writings of competent observers. Ross (1912)
quotes the figures of Welch (1897) a * 6 to 20 or more spores. Later,
however, from observations made by himself and one of us, he
taught that the maximum number was 2 5 or 32. Marchiafava,
Bignami and Mannaberg (1894) state that the numbers vary from
8 to 15, while the following authors give the numbers as follow?.
Stephens and Christophers (1908) 8 to 10, Deaderick (1910) 5 to 25
and even 30, Gulland and Goodall (1912) 8 to 15. It is probable
that these discrepancies are due to two reasons: (a) observations
of autopsy smears in which the sporulation had not reached its full
maturity, and ( b ) observations on autopsy smears of patients to
whom quinine had been given before death. In such cases
incomplete and atypical sporulation is seen. We do not claim,
5 20
SUMMARY
(1) The malignant tertian parasite has been successfully
cultivated, after the method of Bass and Johns, on twelve occasions,
and the benign tertian parasite on three occasions by the present
writers.
(2) It is unnecessary to remove the leucocytes from the blood
before incubation. The optimum temperature would appear to be
about 38° C., and the parasites may grow successfully at a
temperature of 36° C. or 37 0 C.
(3) The time required for the full development of the parasite
in vitro varies, but this variation is partly due to the age of the
parasite at the time of incubation.
(4) The cultures of benign tertian differed from those of
malignant tertian in that there was no tendency to clumping of
the parasites in the former, either before or during sporulation.
(5) This difference appears to us to explain in a satisfactory
manner why only young forms of malignant tertian are found in
the peripheral blood, as the clumping tendency of the larger forms
causes them to be arrested in the finer capillaries of the internal
organs. It also explains the tendency to pernicious symptoms, such
as coma, in malignant tertian malaria. All stages of the benign
tertian parasite are found in the peripheral blood, and there are
seldom pernicious symptoms, because there is no tendency to
clumping.
(6) The malignant tertian parasite ( P . falciparum) is capable of
producing, in maximum segmentation, thirty-two spores. On the
other hand, benign tertian (P. vivax) produces, as a rule, during
maximum segmentation, sixteen spores; sometimes more may be
produced, but the number is never thirty-two.
(7) The pigment in P. falciparum collects into a definite,
circular, and very compact mass early in the growth of the parasite.
On the other hand, during the growth of P. vivax the pigment
remains scattered in definite granules throughout the body of the
parasite, till just before segmentation, when it collects into a loose
mass of granules in the centre of the full-grown Plasmodium .
(8) The morphology of P. falciparum and P. vivax in the human
host is identical with the morphology of these parasites, as obtained
in the culture tube.
5*9
arrested in the internal organs during sporulation, although when
full grown they are much larger than the malignant tertian parasite.
The absence of clumping in the case of the benign tertian parasite
explains satisfactorily the lack of pernicious symptoms in this
infection, and, hence, the absence of comatose malaria, and it also
explains why all stages of this parasite are found in the peripheral
blood, even up to sporulation.
THE MORPHOLOGY OF THE BENIGN TERTIAN PARASITE
IN THE HUMAN HOST
This is identical with that which we have just described under
morphology in the culture tubes (Plate XXXIV, figs. 21-30). The
chief features in which this parasite differs from P. falciparum , are
the large size of the containing corpuscle with the presence of
Schiiffner’s dots, the straggling form of the medium-sized parasites,
the scattered pigment in the pre-segmenting stages and the smaller
number of spores. These spores are larger than the spores of the
malignant tertian parasite. The pigment in the sporulating forms
is collected into a loose mass. With regard to the number of spores
produced by this parasite, we have found as many as twenty-four,
but the most usual number is 16 to 18. In culture the largest
number we obtained on three occasions was 18. The following
are the numbers of spores produced by P. vivax in the
human host, according to several observers. Ross (1910) quotes
the figures of Grassi and Feletti, 15 to 20 spores. Marchiafava,
Bignami and Mannaberg (1894), *6 spores. Golgi quotes 14 to 19
spores. Stephens and Christophers (1908), 15 or more spores.
Deaderick (1910) gives 12 to 26 spores, most often 16 spores. In our
opinion Deaderick’s figures are most correct. All stages of this
parasite are found in the peripheral blood, and the phenomenon
of clumping has never been observed. Ross has taught recently
that there are four splits with this parasite—that is, sixteen spores ;
three splits with the quartan parasites—that is, eight spores; and
five splits, or thirty-two spores, with the malignant parasite.
520
SUMMARY
(1) The malignant tertian parasite has been successfully
cultivated, after the method of Bass and Johns, on twelve occasions,
and the benign tertian parasite on three occasions by the present
writers.
(2) It is unnecessary to remove the leucocytes from the blood
before incubation. The optimum temperature would appear to be
about 38° C., and the parasites may grow successfully at a
temperature of 36° C. or 37°C.
(3) The time required for the full development of the parasite
in vitro varies, but this variation is partly due to the age of the
parasite at the time of incubation.
(4) The cultures of benign tertian differed from those of
malignant tertian in that there was no tendency to clumping of
the parasites in the former, either before or during sporulation.
(5) This difference appears to us to explain in a satisfactory
manner why only young forms of malignant tertian are found in
the peripheral blood, as the clumping tendency of the larger forms
causes them to be arrested in the finer capillaries of the internal
organs. It also explains the tendency to pernicious symptoms, such
as coma, in malignant tertian malaria. All stages of the benign
tertian parasite are found in the peripheral blood, and there are
seldom pernicious symptoms, because there is no tendency to
clumping.
(6) The malignant tertian parasite ( P . falciparum) is capable of
producing, in maximum segmentation, thirty-two spores. On the
other hand, benign tertian (P. vivax) produces, as a rule, during
maximum segmentation, sixteen spores; sometimes more may be
produced, but the number is never thirty-two.
(7) The pigment in P. falciparum collects into a definite,
circular, and very compact mass early in the growth of the parasite.
On the other hand, during the growth of P. vivax the pigment
remains scattered in definite granules throughout the body of the
parasite, till just before segmentation, when it collects into a loose
mass of granules in the centre of the full-grown Plasmodium .
(8) The morphology of P. falciparum and P. vivax in the human
host is identical with the morphology of these parasites, as obtained
in the culture tube.
REFERENCES
Bass and Johns (1912). The Cultivation of Malarial Plasmodia ( Plasmodium vtvax and (
Plasmodium falciparum) in vitro. Joum. Exper. Med., XVI, pp. 567-579.
Cropper, J. (1908). Phenomenal Abundance of Parasites in a Fatal Case of Pernicious Malaria.
Lancet, July 4th, 1908.
Deaderick (1910). Malaria.
Gulland, G. L., and Goodall, A. (1912). The Blood : a guide to its Examination and to (
the Diagnosis and Treatment of its Diseases. (William Green & Sons, Edinburgh and
London.)
Jouxorr, N. M. (1913). Culture du Parasite de la Malaria. Comp. Rend. Soc. Biol., Vol. \ *
LXXIV, No. 3, pp. 136-138.
Marchiafava, E., Bignami, A., and Mannaberg (1894). Two Monographs on Malaria and
the Parasites of Malarial Fever. (1) By Marchiafava and Bignami; (2) by Mannaberg.
Ross, Sir Ronald (1910). The Prevention of Malaria, p. 89.
Rowley- Lawson, Mary (1913). The Extracellular Relation of the Malarial Parasite to the
Red Corpuscles and its Method of Securing Attachment to the External Surface of the
Red Corpuscle. Joum. Exper. Med., Vol. XVII, No. 3, pp. 324-335. With 6 plates.
Stephens, J. W. W., and Christophers, S. R. (1908). The Practical Study of Malaria, p. 34.
Thomson, J. G., and McLellan, S. W. (1912). The Cultivation of one generation of Malarial
Parasites (. Plasmodium falciparum) in vitro, by Bass’s method. Ann. Trop. Med. and
Parasitol., VI, pp. 449-462. Two plates.
Thomson, J. G., and Thomson, D. (1913). The Cultivation of one generation of Benign
Tertian Malarial Parasites ( Plasmodium vivax) in vitro, by Bass’s method. With 1 plate.
Ann. Trop. Med. and Parasitol., Vol. Vll, No. 1, March, pp. 153-165.
Thomson, J. G., and Sinton, A. (1912). The Morphology of Trypanosoma gambiense and
T rypanosoma rbodttitnst in Cultures : and a Comparison with the Developmental Forms,
described in Glossina palpalis . Ann. Trop. Med. and Parasitol., Vol. VI, pp. 331-356
Three Plates.
522
EXPLANATION OF PLATES
Plate XXXIV
All the figures in this plate represent the growth of the parasites
in the culture tube. The figures were drawn with an Abbe camera
lucida from stained preparations. Magnification 1,600 diameters.
Fig. i is a malignant tertian parasite at the time of inoculation
of the culture tube. Corpuscle is shrunken. No
pigment is seen and no stipulation of the corpuscle.
Fig. 2 represents 12 hours’ growth at 36° C. No pigment is yet
visible.
Fig. 3 represents 23 hours’ growth at 41° C. Pigment is not yet
evident, but the parasite has increased in size.
Figs. 4 and 5 represent 36 hours* growth; note the appearance of
a compact mass of pigment. Fig. 5 is a double
parasite.
Figs. 6 and 7 show commencing segmentation after 47 hours’
incubation. Fig. 6 shows five daughter cells, and
fig. 7 twenty spores.
Figs. 8, 9 and 10 show sporulating forms after 51 hours’ incubation.
Fig. 11 shows complete sporulation (32 spores) after 52 hours’
incubation. The containing corpuscle has burst and
liberated the spores.
Fig. 12 was obtained after 56 hours’ incubation.
Fig. 13 represents a young merozoite of the second generation
which has entered a new corpuscle after 75 hours’
incubation. Note that there is no vacuole. The
parasite is only 1 'S/u in its longest diameter.
Fig. 14 represents sporulation of the second generation after
3 days’ incubation.
Fig. 15 shows a young merozoite of the third generation after
4^ days’ incubation.
5 2 3
Fig. 16 represents sporulation in the third generation after 6 days*
incubation. Only eight spores have formed.
Fig. 17 is a young merozoite of the fourth generation after 7 days 1
incubation.
Fig. 18. Eight days’ incubation, showing commencing segmenta¬
tion in the fourth generation.
Fig. 19. Nine days’ incubation, showing sporulation in the fourth
generation.
Fig. 20 shows a young merozoite of the fifth generation after
10 days' incubation.
All of the above figures were obtained from one culture tube,
without the removal of leucocytes and without the addition of fresh
serum or corpuscles.
Fig. 2i shows a young benign tertian parasite at the time of
inoculation of the culture tube. The culture was made
at the period when these young rings predominated.
Note the large size of the corpuscle and the Schuffner’s
dots.
Figs. 22, 23 and 24 represent 8 hours’ growth in culture tube at
39 0 C. The parasites have now distinctly increased in
size^ and fine granules of pigment are now seen
scattered in the protoplasm.
Figs. 25-30 show parasites obtained from the culture tube after
20 to 29 hours* incubation. Fig. 25 shows four masses
of chromatin.
Fig. 30 is a female gamete (undivided chromatin and scattered
pigment) found in the culture tube; similar gametes
were found in the blood at the time of inoculation of
cultures, so it is probable that this gamete was one of
these and that it had not developed in the culture.
Note the large size of the corpuscles in the benign tertian as
compared with those in the malignant tertian; also the
Schuffner’s dots and the scattered pigment. The spores also are
larger and fewer in number in the case of the malignant tertian
parasites.
5 H
Plate XXXV
Microphotographs of malignant tertian parasite. Figs, i to 3
are from the same culture of Plasmodium falciparum , and are all
photographed at the same magnification, namely, 1,500 diameters.
Fig. 1. A photograph of the malignant tertian parasite as seen
in the peripheral blood before incubation. No pigment
is seen. The small dark mass is the chromatin.
Fig. 2. The malignant tertian parasite after 12 hours* incubation
at a temperature of 37 0 C. to 38° C. The parasites have
grown to at least twice the size. A circular mass of
compact pigment is now evident. This photograph
illustrates the clumping tendency and seven parasites
are seen grouped together. No division into daughter
cells has yet taken place. The dark circular area is
pigment, and the darker area, towards the margins, is
due to the chromatin.
Fig. 3. The malignant tertian parasite after 25 hours’ incubation
at a temparature of 37 0 C. to 38° C. This photograph
shows well the tendency to clump and the parasites are
now seen at all stages of sporulation.
Fig. 4. Shows two parasites from a smear of the spleen showing
segmentation. Thirty-two spores can be counted in the
one in the centre of the field.
Fig- 5- This is a photograph taken from a coloured plate drawn
by Dr. Cropper and shows the clumping of the red cells
containing pre-segmenting forms of the malignant
tertian parasites. This drawing was made from a smear
of the peripheral blood (vide 1 Lancet,’ July 4th, 1908,
and coloured plate fig. 1). (Magnification about 1,000
diameters).
5 2 3
Fig. 16 represents sporulation in the third generation after 6 days*
incubation. Only eight spores have formed.
Fig. 17 is a young merozoite of the fourth generation after 7 days'
incubation.
Fig. 18. Eight days' incubation, showing commencing segmenta¬
tion in the fourth generation.
Fig. 19. Nine days' incubation, showing sporulation in the fourth
generation.
Fig. 20 shows a young merozoite of the fifth generation after
10 days' incubation.
All of the above figures were obtained from one culture tube,
without the removal of leucocytes and without the addition of fresh
serum or corpuscles.
Fig. 21 shows a young benign tertian parasite at the time of
inoculation of the culture tube. The culture was made
at the period when these young rings predominated.
Note the large size of the corpuscle and the Schiiffner's
dots.
Figs. 22, 23 and 24 represent 8 hours' growth in culture tube at
39 0 C. The parasites have now distinctly increased in
size, and fine granules of pigment are now seen
scattered in the protoplasm.
Figs. 25-30 show parasites obtained from the culture tube after
20 to 29 hours' incubation. Fig. 25 shows four masses
of chromatin.
Fig. 30 is a female gamete (undivided chromatin and scattered
pigment) found in the culture tube; similar gametes
were found in the blood at the time of inoculation of
cultures, so it is probable that this gamete was one of
these and that it had not developed in the culture.
Note the large size of the corpuscles in the benign tertian as
compared with those in the malignant tertian; also the
Schiiffner's dots and the scattered pigment. The spores also are
larger and fewer in number in the case of the malignant tertian
parasites.
5H
Plate XXXV
Microphotographs of malignant tertian parasite. Figs, i to 3
are from the same culture of Plasmodium falciparum , and are aJJ
photographed at the same magnification, namely, 1,500 diameters.
Fig. 1. A photograph of the malignant tertian parasite as seen
in the peripheral blood before incubation. No pigment
is seen. The small dark mass is the chromatin.
Fig. 2. The malignant tertian parasite after 12 hours' incubation
at a temperature of 37 0 C. to 38° C. The parasites have
grown to at least twice the size. A circular mass of
compact pigment is now evident. This photograph
illustrates the clumping tendency and seven parasites
are seen grouped together. No division into daughter
cells has yet taken place. The dark circular area is
pigment, and the darker area, towards the margins, is
due to the chromatin.
Fig. 3- The malignant tertian parasite after 25 hours' incubation
at a temparature of 37 0 C. to 38° C. This photograph
shows well the tendency to clump and the parasites are
now seen at all stages of sporulation.
Fig. 4. Shows two parasites from a smear of the spleen showing
segmentation. Thirty-two spores can be counted in the
one in the centre of the field.
Fig. 5. This is a photograph taken from a coloured plate drawn
by Dr. Cropper and shows the clumping of the red cells
containing pre-segmenting forms of the malignant
tertian parasites. This drawing was made from a smear
of the peripheral blood (vide ‘ Lancet/ July 4th, 1908,
and coloured plate fig. 1). (Magnification about 1,000
diameters).
Antals Trop. Parasitol., Pol. Vll
PLATE XXXT
3 4
SPORULATION OF THE MALIGNANT TERTIAN PARASITE
ctq photographs by Thomson,]. G. atul D.
C. Tinling Co., Ltd., Imp.
5 2 S
THE TRICHONOCARDIASES
BY
ALBERT J. CHALMERS, M.D., F.R.C.S., D.P.H.,
DIRECTOR, WELLCOME TROPICAL RESEARCH LABORATORIES,
AND
CAPTAIN W. R. O'FARRELL, R.A.M.C.,
PROTOZOOLOGIST, WELLCOME TROPICAL RESEARCH LABORATORIES, KHARTOUM
(Received for publication 27 September , 1913)
Plates. XXXVI—XXXVII
INTRODUCTORY
The literature relating to the Trichonocardiases being of such
recent date and of such small bulk, together with the fact that this
is the first record of their occurrence in the Anglo-Egyptian Sudan,
and, as far as we know, in Africa, has induced us to bring forward
the following observations.
HISTORICAL
A number of affections of hairs have been described which are
characterized by the presence of variously coloured nodes, nodosities
and more or less elongated unilateral, bilateral or annular
thickenings on hairs in various parts of the body. The nodes are
confined to the shaft, and do not penetrate into the hair follicle.
These affections may be classified as the Trichosporoses, Lepothrix
and the Trichonocardiases.
Trichosporosis was first described as a rare disease of the hair
of the head in native women living in the valleys of the province
of Cauca on the slopes of the Eastern Cordilleras of Colombia. It
has been observed in Rio de Janeiro and Germany. It also occurs
on the hair of the beard in men. Osorio gave it the name * Piedra,'
because of the stony hardness of the nodules, but the more correct
name is Trichosporosis tropica, as its causal organism is Tricho -
sporum giganteum , Behrend, 1890. A similar condition has been
526
briefly described by Castellani as occurring in India and Ceylon,
but so far, he has not named the fungus, which appears to differ
from T. giganteum.
The remaining Trichosporoses belong to temperate climates; they
are: —
Biegel’s Trichosporosis, or Tinea nodosa of Cheadle and Morris,
due to Trichosporum beigeli (Rabenhorst, 1867). It was first found
by Lindemann in 1865, and is often called the Chignon fungus.
It occurs on the hair of the head.
Unna’s Trichosporosis, or Piedra nostras, found on the hairs of
the beard and moustache, and due to T. ovale (Unna, 1896).
Behrend’s Trichosporosis, or Nodular trichomycosis, found on
the hairs of the beard, and due to T. ovoides (Behrend, 1890).
Du Bois’ Trichosporosis, found on the hairs of the pubic region,
and due to T. glycophile (Du Bois, 1910).
Another nodular hair disease is Lepothrix, or scaly hair, which
was discovered by Paxton in 1869, was named by Wilson, and
described by Eisner and Crocker. It is characterized by variously
coloured grey, yellow, or brown nodules, which surround the hairs
of persons who perspire profusely. The hairs most usually
attacked are those of the axillae, but the disease may also be found
on those of the chest, pubes, and inside of the thighs. It is
common in Germany, and is believed to be due to a diplococcus
forming zoogloea masses. Allied to Lepothrix is the affection called
Trichomycosis capillitii by Winternitz in 1903, which is said to be
due to a straight or slightly bent bacillus. The descriptions of
these conditions suggest that there may be some confusion, and that
in reality they may be Trichonocardiases in which either only the
diplococcus has been observed or in which only the bacilliform
fungus has been seen. In any case it would seem that the disease
Lepothrix requires further investigation, especially as Crocker’s
excellent description closely agrees with Trichonocardiasis.
The Trichonocardiases are allied in appearance to Lepothrix,
but differ therefrom in the fact that they are all due to a fungus
Nocardia tenuis (Castellani, 1911), either alone as in Trichono¬
cardiasis flava (PI. XXXVI, figs. 1 and 2; PI. XXXVII, fig. 13),
or in association with a Micrococcus as in Trichonocardiasis nigra
(PI. XXXVI, figs. 4 and 5; PI. XXXVII, figs. 14, 15 and 20), and
5 2 7
T. rubra (PI. XXXVI, figs. 6 and 7; PI. XXXVII, figs. 16, 19 and
22). They were first described by Castellani in 1911 in a paper
published in the British Journal of Dermatology, and this description
was extended by the same author in 1912 in an article published in
the Transactions of the Royal Society of Medicine, and still later,
in 1913, in the second edition of the Manual of Tropical Medicine
published by himself and one of us. He named the disease
Trichomycosis axillaris, and sub-divided it into three varieties,
flava, rubra and nigra; but as it occurs on the hairs of the pubes,
in some cases without any axillary infection, we are of the opinion
that Trichonocardiasis is a better term.
Attention has, very briefly, been drawn to Castellanos work by
Jackson and McMurty in their book on ‘Diseases of the Hair,*
published in 1913, where they devote a small section to the subject
under the title ‘ A Disease resembling Lepothrix.’
So far as we know, the above comprise the total literature on
the subject of the Trichonocardiases.
GEOGRAPHICAL DISTRIBUTION
We have found the three varieties of Trichonocardiasis in the
Anglo-Egyptian Sudan, and one of us has observed a condition
resembling Trichonocardiasis flava in Europeans on the Gold Coast,
West Africa, as long ago as 1898.
As the three varieties may be found on one and the same person,
and, indeed, in the same axilla, we do not propose to give separate
descriptions of each variety, but to describe the disease Trichono¬
cardiasis as a whole, mentioning the difference between the three
varieties when necessary.
At present the Trichonocardiases are only known to exist in the
Tropics, i.e., in Ceylon, the Anglo-Egyptian Sudan, and on the
Gold Coast. These regions being so widely separate one from the
other, it is reasonable to suppose that the disease will be found in
other tropical regions when looked for.
Castellani has drawn special attention to the fact that he found
his cases in the hot, damp districts of Ceylon, and adds that it
will probably be found occurring in every low-lying tropical region.
While our Gold Coast cases support Castellani’s views, the cases we
528
have found in Khartoum demonstrate that the disease may also
occur in a hot and dry climate and at a certain elevation. We found
our first cases here in the month of May of this year, when the
highest shade temperature was II2'2°F. (44*5° C.), and the mean
shade temperature was 105*9° F. (41° C.). During this time the
least percentage humidity of the air was 4 % and the average 21%,
while the evaporation from Piche's tube rose as high as 30*1 milli¬
metres. As the elevation of Khartoum is 390 metres above sea-level
at Alexandria, it is not low-lying. We, however, agree with
Castellani that the disease will probably be found in many tropical
regions, low-lying or elevated, if looked for.
RACIAL DISTRIBUTION
Most of our cases have occurred in Europeans, but Castellani
has seen it among the Sinhalese and we have met with it in a negro.
Possibly its occurrence was noticed long ago by native doctors, and
may be one reason why so many native races shave the axillae.
BODY DISTRIBUTION
So far we have only seen the infection on the hairs of the axillae,
pubic regions, and folds of the groin and in this we are entirely in
agreement with Castellani.
The reason for the distribution appears to us to be the nutrition
of the causal fungus Nocardia tenuis , as it would appear to require
a mixture of human proteid material such as is found in blood
serum and normal saline before it will sprout. As we shall point
out later, it derives its nutriment from the lymph of the cortex of
the hair while at the same time it is bathed by the saline sweat of
the axillae, pubic regions, and groins, and this appears to be the
reason of its affinity for these particular parts of the body.
AETIOLOGY
The causal agent is 'Nocardia tenuis (Castellani, 1911) (PI.
XXXVI, fig. 9; PI. XXXVII, fig. 17), either alone as in the variety
Trichonocardiasis flava, or associated with Micrococcus nigrescent
(Castellani, 1911) as in Trichonocardiasis nigra, or with Micrococcus
castellanii (Chalmers and O’Farrell, 1913) (PI. XXXVII, fig. iS N
as in Trichonocardiasis rubra.
We will now consider these aetiological factors seriatim.
529
NOCARDIA TENUIS (Castellani, 1911). This organism can
be readily seen in cleared specimens of the infected hairs mounted in
gelatin or Canada balsam as minute rod-like or bacillary forms
often lying more or less parallel to one another and embedded in
an amorphous ground substance (PI. XXXVII, fig. 22), probably
excreted by the fungus. This ground substance, as will be explained
later, is fixed to the cortex of the hair, and lies under the cuticle
and some of the superficial layers of the cortex.
When individual hyphae are examined after coloration by any
of the ordinary stains, they are seen to be narrow, elongated,
unbranched, non-septate rods resembling bacilli (PI. XXXVI,
fig- 9; PI- XXXVII, fig. 17). They may be short or long, straight
or slightly curved, but they are always fairly and usually very
narrow.
A series of measurements gives the average length as varying
from about 2 0 to over 7*0/1, and the average breadth from 014
to 0*3/1.
Each hypha is enclosed in a cell wall, inside which lies the
cytoplasm, which may stain very lightly with the colouring agent
used, but may also stain fairly darkly, especially in certain areas.
No nucleus can be demonstrated and the cytoplasm appears to be
homogeneous except at certain places where well-defined deeply
staining rounded granules may be seen (PI. XXXVI, fig. 9). These
granules, which measure about 0*14 to 0*3/1 in transverse diameter,
may be situated at one end of the hypha, as in PI. XXXVI,
&gs. 9, 10, 12, and PI. XXXVII, fig. 21, or this may be placed at
regular intervals through the cytoplasm (PI. XXXVI, fig. 9), or one
may be at one end while others may be close together near the
opposite pole. We have not met with any branched forms, except in
cultivation, but we have seen forms with one end rather bulbous,
resembling in miniature the club forms of some other Nocardias.
We have not succeeded in cultivating this fungus permanently, in
bulk, or pure cultivation, but we have succeeded in inducing growth
by placing scrapings from the hairs in equal parts of human blood
serum and 0*85% normal saline solution in hanging drop prepara¬
tions. These were first kept for twenty-four hours at the room
temperature, which varied from 90° F. (32*2° C.) at night to about
105° F. (40*6° C.) in the day, and were then placed in an incubator
at 104° F. (40° C.).
53°
Under these conditions the fungus quickly grew out of its
amorphous bed (PI. XXXVI, fig. io) into the surrounding liquid,
and proceeded to develop in the manner depicted in PI. XXXVI,
fig. 12, and PI. XXXVII. fig. 21, the latter of which is a
m icrophotograph.
It will be noticed that the branching is monopodial, and that
the hyphae are non-septate.
With regard to its position in the vegetable kingdom it is obvious
that, as it possesses neither chlorophyll nor chromatophores, it must
belong to the class Fungi, and that it belongs to the fungal order
Deuteromycetes or Fungi imperfecti, because its life-cycle is
unknown. Further, it can be placed in the sub-order Hyphomycetes
because perithecia, asci and immersed stroma are also unknown. Of
the families of the Hyphomycetes it belongs to that of the
Mucedineae because its hyphae are pallid and do not cohere to
form a compound stem-like structure. Lastly, it may be grouped
with other forms in the genus Nocardia (Toni and Trevisan, 1899),
which is defined as consisting of mycelial filaments of various sizes,
but generally very thin (one micron or less in diameter), often
branching, non-septate, and without differentiated nuclei.
Nocardia tenuis (Castellani, 1911) may therefore be defined as
follows : —
Nocardia composed of thin, bacillary-like hyphae, varying from
about 2 to over 7/1 in length, and from o’14 to o’3/1 in breadth,
embedded in an amorphous ground substance attached to the cortex
of the axillary and pubic hairs of man in tropical countries.
MICROCOCCUS NIGRESCENS (Castellani, 1911). This
coccus has been grown in pure culture by Castellani, who has
described its biological characters on page 1521, and has illustrated
its cultural appearances in fig. 8 on Plate V of the second edition
of the Manual of Tropical Medicine mentioned above. It is there¬
fore unnecessary for us to go into details with regard to this
organism, except to say that Trichonocardiasis nigra appears to be
much rarer here than the flava or rubra, while the pigmentation on
the hairs and in the growths we have obtained is relatively scanty,
and we have found it difficult to isolate and grow Micrococcus
castellanii (Chalmers and O’Farrell, 191.3). Castellani in Ceylon
found that it was more difficult to isolate and grow this coccus than
was the cr.se with Micrococcus nigresccns. On the other hand, we,
S3i
in Khartoum, have found quite the reverse: we have found it
exceedingly difficult to obtain the black coccus with anything beyond
a trace of black pigment, while the red coccus grows easily and well.
It is very readily separated out by sowing scrapings from the
hair, or pieces of the hair on sloped agar, and picking out either
the red colonies which require some time to develop the reddish
pigment, or the yellow colonies which always appear first and in
which the red pigment subsequently shows itself.
MICROCOCCUS CASTELLANII is a round or oval coccus,
measuring from about 0 3 to about 07 fi in diameter (PI. XXXVII,
fig. 18). It is divided medianly by a clear central line into two
half-moon shaped segments, thus producing the well-known
diplococcal appearance. The cocci, which may occur singly or in
groups, are colourless and non-motile, but excrete an amorphous
non-granular lemon chrome-coloured pigment (classified according
to Ridgway’s Colour Standards), which lies between the individual
cocci.
In old cultures another pigment of dark brownish-red colour
appears (according to Ridgway’s Standards this colour is madder-
brown), but its relationship to the earlier yellow pigment can easily
be proved by removing some of it and suspending in a sufficiency of
distilled water, when the fluid at once resembles a similar suspension
of the yellow pigment. If, however, this is done in a very small
quantity of water, a faint reddish tinge can be observed. The best
medium for showing the striking yellow pigment is the potato,
when in twenty-four hours the growth assumes the appearances
depicted on PI. XXXVI, fig. 11. In order to demonstrate the red
pigment it is necessary to cultivate the microccocus for some time.
The medium on which this pigment shows best is the ordinary agar
slope, which, when old, exhibits the dark-red pigment in the centre
and the yellow pigment at the sides (PI. XXXVI, fig. 8). The yellow
pigment is seen both in aerobic and anaerobic growths. In broth
and peptone water or liquid blood serum, whether grown aerobically
or anaerobically, the pigment is not visible until a very abundant
growth appears, when the yellow colour can be seen.
The coccus colours with all the ordinary staining re-agents, and
as a rule is decidedly Gram positive. It must, however, be admitted
that even in preparations showing most of the cocci well coloured
532
when treated by Gram’s methods, a few may be seen quite
decolourised, and we have also seen cocci with one demi-lunar
segment well stained while the other was completely decolourised.
We have failed to demonstrate a capsule around the coccus.
The organism grows aerobically and also anaerobically. The
optimum temperature appears to be 37° C., but it can also be
cultivated at the room temperature, which in Khartoum at this
season of the year is seldom less than 32'2° C. It also grows at
20 0 C. on agar slopes, but not as abundantly as at 37° C., and is
therefore less pigmented. Its rate of growth depends somewhat
upon the medium, as it grows quickest and best on potato, and next
best on ordinary or glycerin agar. On solid media it gives rise to
a yellow growth at first; but on most agar media, if kept long
enough, some red coloration will subsequently be found. The best
medium for the red coloration appears to be the ordinary agar
slope. With regard to the other agar media, it grows well on
glucose and maltose agar. Like Micrococcus nigrescens, it produces
neither acid nor gas in the following media: —
Peptone-sugars : Glucose, Laevulose, Galactose, Arabinose,
Maltose, Lactose, Saccharose, and Raffinose.
Peptone-carbohydrates-. Dextrin and Inulin.
Peptone-glue0side : Amygdalin.
Peptone-alcohols: Erythrite, Adonite, Dulcite, Isodulcite,
Mannite, Sorbite, and Inosite.
It grows slowly on blood serum, which it does not liquefy, and
well in broth and peptone water, in which it forms a general
turbidity. It does not produce indol.
In agar stabs the growth is confined to the line of puncture and
to the formation of a small yellow knob on the surface, in other
words, the growth is filiform.
With regard to its systematic position in the family Coccaceae
(Zopf, emended Migula), it must certainly be grouped with the
genus Micrococcus (Hallier, 1866, emended Cohn, 1872).
In this genus it certainly belongs to those forms which grow
well on agar media and are Gram positive, and in this division it
belongs to the sub-division which produces colours.
The cocci of this sub-division which possess red coloration are
533
only three in number, Micrococcus roseus (Bumm, 1885), Micrococcus
ruber (Trommsdorff, 1904), and Micrococcus rubidus (Hefferan).
Under the term M. roseus (Bumm, emended Lehmann and
Neumann) are gathered a large number of rose coloured diplococci
which are not known to be parasitic, and which produce growths on
potato which are limited to the streaks. These growths are faint
rose red in colour within an oily lustre, and are often surrounded by
a whitish glistening zone, thus giving rise to a very different
appearance from that produced by M . castellanii . Micrococcus
ruber (Tromsdorff, 1904) or, as it is sometimes named, M. chro-
midrogenus ruber , which was isolated from axillary hairs, is
characterised by the fact that it does not grow on potato, its
colouring matter is not soluble in water, and when treated by
sulphuric acid the red colour turns blue-green, while M. castellanii
does grow on potato and its colouring matter is unaffected by
25 per cent, sulphuric acid. Moreover, M. ruber is associated with
chromidrosis, while M. castellanii is not. With regard to Micro¬
coccus rubidus , it is impossible to obtain any information as to its
characters in Khartoum, but it appears to be closely related to
M . roseus var carneus , and to be non-parasitic.
We, therefore, are of the opinion that the organism which we
are describing is not the same as any species of Micrococcus so far
named, and therefore give it the appellation Micrococcus castellanii ,
Chalmers and O’Farrell, 1913.
METHOD OF INFECTION
There is no doubt in our minds that the usual method of infection
is from man to man, and we base this opinion upon the following
observation: —
A young European, known to be quite uninfected with these
parasites, became greatly interested in our work; he examined the
cases which came to the laboratory, and handled and examined the
fresh hairs removed from the axillae.
When he visited the laboratory he was clad in tennis attire, and
as he moved about, his shirt rucked up from below repeatedly, and
whenever this happened he adjusted it by pulling forward his
leather belt with one hand while he pushed the soft shirt downwards
inside his trousers with the other. Two weeks after examining the
534
cases he first noticed a reddish appearance on the hairs of the pubes.
On examination it was found that the hairs were infected with the
red variety of Trichonocardiasis, that is to say, with the variety
which he saw and handled in abundance. It grew rapidly on the
pubic hairs, but did not extend to the axillae. Moreover, it only
grew in the fold of the left groin, where sweat is apt to accumulate.
It did not spread to the right groin.
INCUBATION PERIOD
It would therefore appear as though two weeks were required to
elapse before an infection produces sufficient growth on the hairs
to attract attention, even when it is being looked for.
PATHOLOGY
The pathology of the infection appears to be quite simple.
The fungus arriving upon the hair grows and pushes its way
at first under a cuticular scale and then quickly works its
passage into the cortex, raising, in so doing, its superficial fibres,
which, together with the cuticular scale, form a covering or pro¬
tection for the fungus (PI. XXXVII, fig. 19), which probably finds
the nutriment it requires in the fluids of the cortex of the hair.
Once established in position the fungus does not penetrate deeper
into the cortex, but, on the contrary, grows outwards, forming the
nodosities, rings and sheaths to the hair which have already been
mentioned. It is owing to this method of growth that the hair
is so little affected, as the deeper layers of the cortex, the medulla
and the root of the hair are not involved in our cases.
MICROSCOPICAL ANATOMY
In an early stage of the infection the cuticle and some fibres of
the cortex (PI. XXXVII, fig. 13, shows this stage at three places
between the large clumps of fungus) may be seen to be raised in
ridges, which run in wavy lines across the shaft of the hair
transversely to its long axis. Under these ridges masses of the
fungus may be seen growing.
A parasitic mass is composed of a firm homogeneous ground
substance, in which the rod-like hyphae of the Nocardia may be seen
535
(PI. XXXVII, fig. 22). In this the cocci are also found which are
responsible for the coloration of the black and red Trichono-
cardiases.
SYMPTOMATOLOGY
As already stated the shortest incubation period with which we
are acquainted is two weeks. The infection causes no general
symptoms and is only discovered by accident by the patient, who
then seeks advice in order to know what is taking place. Very often
the infectiQn is first met with while the patient is being examined
by his medical attendant for some other disease. In regiments it
is usually found during a general medical inspection.
The regions of the body which are affected are the axillae and
the pubes, most commonly the former alone or in conjunction with
the latter, and less commonly only the latter. The hairs of these
regions may be affected with one or more of the three varieties
already mentioned.
On examining with the naked eye an axilla affected by the yellow
variety, it will be noticed that the hairs are covered with a thick
or thin yellow deposit (PI. XXXVI, fig. 1), as though the natural
grease had accumulated, and, indeed, this is the reason why some
persons ask about the complaint, as they consider that it reflects upon
their habits. If the black variety is present it will appear as black or
very dark patches, short or long, thick or thin, on one side of or
ensheathing the hair (PI. XXXVI, fig. 5), and the same description
holds good for the red variety (PI. XXXVI, fig. 7), with the
exception of the difference in colour. These observations are easily
confirmed by microscopical examination, when the masses may
be seen encircling the hair, lying on both sides or merely forming
excrescences on one side of the shaft. The cuticle and fibres of the
cortex may be seen covering the small masses or embedded in the
large ones.
As already stated, the disease is confined to the shaft of the
hair, the health of which it does not affect. The infection of the
hair does not appear in any way associated with excessive sweating,
though, of course, the amount of this excretion produced by the
axillae is considerably greater in the tropics than in the temperate
zone, and, therefore, in warm climates the axillae are generally
536
damp and the pubic regions are also damper than usual. Probably
it is the dampness of these areas of the body which determines
the distribution of the infection, as it has so far never been seen
on the hairs of the head, face, arms and legs.
The infection dies away on return to a temperate climate,
especially in winter.
DIAGNOSIS
The appearance of minute nodules, rings or sheaths of a yellow,
black or red colour on the hairs of the axillae or pubes indicates
the presence of either Lepothrix or Trichonocardiasis.
The latter may be differentiated from the former by demon¬
strating the presence of Nocardia tenuis , Castellani, 1911, which is
absent in Lepothrix.
From Trichosporosis tropica or Piedra it may be distinguished
by the fact that it does not attack the hair of the head or face, and
also by the difference in the causal parasite. From the other
Trichosporoses it may be separated by its distribution and by the
presence of 'Nocardia tenuis , as well as by the absence of a
Trichosporum.
PROGNOSIS
It appears to have a marked tendency to remain in the region or
regions which it first invaded, and does not, as far as our
observations go, tend to spread over the body.
TREATMENT
We have found the treatment recommended by Castellani to be
quite useful, viz., a lotion of one drachm of formalin to one ounce
of water applied twice daily to the affected hairs, while a 2% sulphur
ointment is used at night.
PROPHYLAXIS
The infection appears to be only mildly contagious; still it does
tend to spread in communities. Cases which are likely to be sources
of infection should have their clothes disinfected before being
washed. During the infection the underclothing should be dusted
with some antiseptic powder, e.g., dermatol, one drachm in one
ounce of Venetian talc, or with a boracic acid dusting powder.
537
Acknowledgments
We wish to acknowledge with many thanks the kindness received
from Major Forrest, R.A.M.C., during the preparation of this
paper, and the skill with which our artist, Mr. A. Marshall, Senior
Laboratory Assistant in these laboratories, has prepared the
illustrations.
Khartoum,
September i $tk, 1913.
REFERENCES
(Arranged in chronological sequence.)
Castellani (1911). British Journal of Dermatology, XXIII, p. 341.
Castellani (1912). Transactions of the Royal Society of Medicine, VI, No. 2, Dec., p. 23.
Castellani, and Chalmers (1913). Manual of Tropical Medicine, and ed., pp. 820 and
1521, Plate V, fig. 8.
Jackson and McMubty (1913). Diseases of the Hair, p. 294.
538
EXPLANATION OF PLATES.
Plate XXXVI
Fig.
1.
Trichonocardiasis flava.
Natural size.
Fig.
2.
Trichonocardiasis flava.
x 60.
Fig.
3 -
Trichonocardiasis flava.
x 40.
Fig.
4 -
Trichonocardiasis nigra.
x 60.
Fig.
5 -
Trichonocardiasis nigra.
Natural size.
Fig.
6 .
Trichonocardiasis rubra.
x 20.
Fig.
7 -
Trichonocardiasis rubra.
Natural size.
Fig.
8 .
Micrococcus castellanri;
Natural size.
old culture on an agar slope.
Fig.
9 -
Nocardia tenuis-, separate hyphae. x 1,200.
Fig.
10.
Nocardia tenuis sprouting out from the ground-substance;
hanging drop preparation, x 1,500.
Fig.
11.
Micrococcus castellanii ;
Natural size.
24 hours’ growth on potato.
Fig.
12.
Nocardia tenuis \ growing in hanging drop preparation.
X 1,500.
538
EXPLANATION OF PLATES.
Plate XXXVI
Fig.
1.
Trichonocardiasis flava.
Natural size.
Fig.
2.
Trichonocardiasis flava.
x 60.
Fig.
3 -
Trichonocardiasis flava.
x 40.
Fig.
4 -
Trichonocardiasis nigra.
x 60.
Fig.
5 -
Trichonocardiasis nigra.
Natural size.
Fig.
6.
Trichonocardiasis rubra.
x 20.
Fig.
;•
Trichonocardiasis rubra.
Natural size.
Fig.
8.
Micrococcus castellanti;
Natural size.
old culture on an agar slope.
Fig.
9 -
Nocardia tenuis-, separate hyphae. x 1,200.
Fig.
10.
Nocardia tenuis sprouting out from the ground-substance;
hanging drop preparation, x 1,500.
Fig.
11.
Micrococcus castellanii ;
Natural size.
24 hours* growth on potato.
Fig. 12. Nocardia tenuis ; growing in hanging drop preparation,
x 1,500.
. hmo/.s Trap. Med. d Paraeifol., Vo/. IIJ
Plate XXXV!
THE TRICHONOCARDIASES
hreP7t,ht4 fo/ftn fan SCrtce
540
PLATE XXXVII
Microphotographs of the Trichonocardiases.
Fig. 13. Trichonocardiasis flava. x 60.
Fig. 14. Trichonocardiasis nigra, x 60.
Fig. 15. Trichonocardiasis nigra, x 60.
Fig. 16. Trichonocardiasis rubra, x 60.
Fig. 17. Nocardia tenuis, Castellani, 1911. x 1,200.
Fig. 18. Micrococcus castellanii , Chalmers and O'Farrell, 1913.
x 1,500.
Fig. 19. Hair showing the fungal growth elevating the cuticle and
the superficial fibres of the cortex, x 500.
Fig. 20. Trichonocardiasis nigra, x 60.
Fig. 21. Nocardia tenuis growing in a hanging drop preparation,
x 1,200.
Fig. 22. Fresh preparation of a hair affected with Trichono¬
cardiasis rubra, showing the radiating hyphae at the
periphery and the optical section of the hyphae in the
centre, x 700.
I
Annals Prop. M:d. & Parasilol.j Vol. Vll
PLATE XXXVII
THE TRICHONOCARDIASES
5+i
EPIDEMIC TRICHONOCARDIASIS
BY
ALBERT J. CHALMERS, M.D., F.R.C.S., D.P.H.
DIRECTOR, THE WELLCOME TROPICAL RESEARCH LABORATORIES
AND
Captain A. D. STIRLING, R.A.M.C.
SENIOR MEDICAL OFFICER, BRITISH TROOPS, KHARTOUM
(Received for publication 25 November , 1913)
INTRODUCTORY
The literature connected with Trichonocardiasis being new and
of small extent, it may be of interest to record an epidemic
which recently took place in the Welsh Regiment in Khartoum.
Before commencing the account of the epidemic we may mention
that we have heard of a case of Trichonocardiasis rubra in a
European while in Aden, but whether it originated there or was
merely imported we are not in a position to state.
- It would, however, appear that the Trichonocardiases are widely
distributed throughout the tropics and, further, that they may occur
in epidemics.
Full accounts of the history, aetiology, etc., having been
already given during this year by Castellani, O’Farrell and one of
us, we restrict our remarks entirely to the present epidemic, its
probable origin and its suppression.
THE EPIDEMIC
About the middle of September, 1913, a Private in the
Welsh Regiment, which had been stationed in Khartoum for
some months, applied for medical treatment on account of
irritation of the skin in both axillae. On investigation
it was found that the hairs of these regions were heavily infected
with Trichonocardiasis rubra. Figure I is a photograph of one
of his axillae and shows the remarkably heavy infection of the
54 2
hairs which were matted together. The skin surrounding this mass
of hairs was red, congested, and very irritable. The individual
hairs, separated out and photographed, are depicted, natural size,
in Figure II, which is improved if examined by a lens. As,
however, this affection of the hair has been recently described by
Castellani and by one of us in conjunction with Captain O’Farrell,
R.A.M.C., no further remarks will be made as to the clinical
appearances or the microscopical findings, except to state that they
entirely agree with those already reported.
Fig. I.—Photograph of an axilla, showing
infection with Trichonocardiasis rubra.
Note the thickenings and the matting
of the hairs.
Fig. II.—Photograph of hairs from the axilla
depicted in Fig. I, natural size. This
figure is improved if examined by means of
a lens.
The occurrence of this case led to a systematic investigation of
the regiment, when no less than 42 cases, in all, were discovered.
Some of the men complained of slight irritation in the axillae, and
these men invariably had an area of erythema surrounding the hairs
of the axilla, but the majority of men had felt no symptoms and
did not appear to have noticed the altered colour of the hair. None
of the cases showed any affection of the hairs on the pubis or any
other part of the body.
The most common variety was Trichonocardiasis flava, while a
543
number of cases showed Trichonocardiasis rubra, but no Trichono-
cardiasis nigra could be found.
The erythema noted was only seen in men suffering from
Trichonocardiasis rubra.
A thorough investigation was made as to the possible source
of infection, and very early suspicion was aroused that the washer¬
men might possibly be the spreaders.
There were, in all, five washermen and four washerwomen for
the cleansing of the regimental clothing. All these, save one,
worked in the central washhouse and these were found to be free
from Trichonocardiasis. The remaining washerman, who was
occasionally assisted by one of the other washermen, went from
bungalow to bungalow in the barracks for the purpose of doing
extra washing for the men, as the regulation washing was
insufficient to keep the clothing clean in a hot dusty country like
the Anglo-Egyptian Sudan. This man washed, in particular, the
shirts and the khaki trousers for the men. He was found to be
infected with Trichonocardiasis flava and rubra, and he was
strongly suspected to be the spreader of the infection, as it was not
confined to the men of any one company but scattered through all
the companies of the regiment.
This washerman and the infected men were duly treated, with
the result that during the last inspection, made a few days ago,
no new cases were found.
TREATMENT
As so many men were affected it afforded ample opportunity for
observing the results of various forms of anti-parasitic treatment.
Most of the cases were treated by means of a lotion of formalin
40 per cent., one drachm to six ounces of rectified spirit. This
lotion was carefully applied to the hair twice daily. It was
advisable to restrict this application entirely to the hairs as it was
often found to irritate the skin if it was carelessly rubbed into the
axilla.
At night a 2 per cent, sulphur ointment was applied, thus
completing the routine recommended by Castellani. The result
was excellent, but it required time, and, on an average, a cure was
not effected under three to four weeks.
544
If the cases did not improve under this treatment, application
of tincture of iodine effected a cure in three to four days.
Tincture of iodine alone was also applied, but with doubtful
advantage.
Perchloride of mercury was also tried, but it required three to
four weeks to bring about a cure.
The best remedy in our hands appeared to be a combination of
the formalin-sulphur method for about two days, followed by
tincture of iodine, when a cure was effected in several cases in less
than a week.
With regard to the erythematous areas mentioned above, these
were readily healed by the application of Calamine lotion,
associated with the anti-parasitic treatment to the hairs, as already
described.
In addition, all the underclothing was dusted regularly with a
boric acid powder of the strength of one drachm to the ounce of
starch powder. This is not the best powder for the purpose, but it
is the cheapest and is readily available. The use of dermatol,
europhen or of xeroform with Venetian talc would be better, but
the boric acid served its purpose quite well.
Khartoum,
November 12/A, 1913.
REFERENCES
Castillani and Chalmers (1913)* Manual of Tropical Medicine, second edition, p. 1525.
Chalmers and O'Farrell (1913). The Trichonocardiascs. Annals of Tropical Medicine and
Parasitology, Vol. VII, pp. 525-540.
545
HEREDITARY INFECTION, WITH
SPECIAL REFERENCE TO ITS
OCCURRENCE IN HYALOMMA
AEGYPTIUM INFECTED WITH
CRITHIDIA HYALOMMAE
BY
Captain W. R. O’FARRELL, R.A.M.C.
I'ROTOZOOLOGIST TO THK WELLCOME TROPICAL RESEARCH LABORATORIES, KHARTOUM
(Received for publication 8 October , 1913)
PLATES XXXVIII—XL
INTRODUCTION
Our knowledge of the role played by the blood-sucking
parasites as transmitters of human and animal diseases has consider¬
ably advanced during recent years. Consequently, the study of
these vehicles of transmission has increased, and many cases of
hyper-parasitism have been discovered. During the search for the
pathogenic varieties others have been encountered, and these, though
their pathogenicity be confined only to their insect hosts, must not
be overlooked.
In three ways, a comprehensive study of their life cycles is of
value: Firstly, in order to prevent their stages from being confused
with the stages in the life histories of more important parasites.
Secondly, to act as a guide to the possible behaviour of the disease-
producing varieties, and thirdly, as a weapon of offence against
their parasitic hosts.
Probably the most important stage in the life cycle of all hyper¬
parasites is the hereditary infection stage. Hereditary infection
does not deal with individuals but with generations, and this
method of infection is a higher* grade of organisation and of evolu¬
tion than simpler methods of infection. The complete life cycles
of but few hereditary infections are known, though a considerable
number of hereditary infection-transmissions are known to exist.
546
Consequently any addition to our knowledge of such infections is
of value. Moreover, any example that can be studied with any
degree of facility is worthy of note. The hereditary infection of
Hyalomma aegyptium by Crithidia hyalommae is one of these, and
will be described in the following pages.
HISTORICAL
The occurrence of hereditary infection has been suggested,
observed and worked out in the insect flagellates already, and
among the authors who advanced this line of research I have
primarily to mention Miss A. Porter, whose work on Crithidia
melophagia y Flu, 1908, has proved of valuable assistance. As
early as 1903 Dr. L. W. Sambon put forward the hypothesis of the
possibility of the occurrence of hereditary infection of the tsetse
fly by Trypanosoma gambiense.
P. C. Flu (1908) published an account of a flagellate parasitic
in the ovaries and small larval forms of the sheep-ked, Melophagus
ovinusy Linnaeus.
L. D. Swingle (1909) working in Nebraska, described rounded
and plasmodial stages of the same parasite, occurring in the egg of
Melophagus ovinus.
A. Porter (1910) described the intestinal contaminative method
of infection and the full cycle and mode of hereditary infection
of the same parasite in the host, egg, developing embryo, and fully
hatched Melophagus .
In a preliminary note published in August, 1913, I have given
a brief description of the flagellate stage and discussed the move¬
ments of Crithidia hyalommae , O’Farrell, 1913. The movements
will not therefore be further described, except the special movement
connected with the entry of the flagellate into the ovarian system of
the tick. The life cycle passed in the larva and nymph will form
the subject of a future communication.
THEORETICAL APPLICATIONS
Apart from the practical proofs of the hereditary transmission
of Crithidia hyalommae in Hyalomma aegyptium , certain theo¬
retical conclusions formed the basis of the search for such proofs.
These conclusions were: —
547
1. The infection was not inoculative , i.e., derived from
the vertebrate host, because: —
(a) All the ticks collected from one given cow were not
infected.
(i b ) Ticks other than Hyalomma aegyptium , although
among batches of infected ticks, never showed infection.
(c) The cows from which the infected ticks were
collected were invariably healthy.
2. The infection was not contaminative , because: —
(a) Ticks, unlike other blood-sucking parasites gener¬
ally, do not move about much on their hosts (except the
males), but settle in one place and slowly secure an attach¬
ment, where they remain till replete.
( b ) The natural secretions and excretions dry very
rapidly, and are often voided on the earth after the tick
detaches itself from its host.
3. The infection was hereditary , because: —
(a) The height of crithidial infection occurred about the
time of oviposition.
( b ) The occurrence of the infection in batches of ticks
all of which probably belong to the same generation.
The practical application of these lines of thought worked out
as follows: —
1. The intestinal diverticula of the ticks were never found to
harbour parasites.
2. The haemocoelic fluid was the habitat of the early pre¬
flagellate forms.
3. These early pre-flagellate forms developed into full grown
Crithidia in the haemocoelic fluid. When these were present
in large numbers they were found to migrate to the ovarian
system.
4. The hereditary infecting forms were found entering the ova.
5. Plasmodial forms were found in the deposited eggs.
MATERIAL AND METHODS
Many specimens of Hyalomma aegyptium were examined from
different localities, but always from cows, which are their common
hosts. Some batches of ticks never showed infection, while in others
infection was the rule.
548
To study the living organism, living specimens were examined
with ordinary light and with dark ground illumination, but with the
latter the Crithidia were found to degenerate and die rapidly
because of the strong light and heat produced by the source of
illumination used in working with a paraboloid condenser. Vital
staining was also employed.
To obtain the haemocoelic fluid a portion of any of the legs is
removed with a fine pair of scissors. For stained specimens of
tissues all ticks were carefully dissected under a dissecting
microscope, and the organs after carefully washing in saline
solution, as recommended by Sir William Leishman, were smeared
out on slides.
Osmic acid and alcohol were employed as fixatives, and also
special methods of wet fixation were used, such as Schaudinn’s
fluid (corrosive-acetic-alcohol), and Bouin’s fluid (picro-formoh
acetic-alcohol).
Various stains were used—Leishman’s and Giemsa’s solutions
gave good results. Bohmer’s haematoxylin (which in this climate
is rather superior to Delafield’s haematoxylin) and Heidenhain’s
iron haematoxylin and its modifications were employed.
Cultivation of the flagellate has proved negative up to the
present, together with attempts to inoculate animals. Frequent
examinations and cultivation of the blood of the cows have given
negative results.
DISTRIBUTION OF THE PARASITE IN THE HOST
The majority of insect flagellates are found parasitic in the
alimentary tracts of their hosts, but in this respect the Crithidia of
Hyalomma aegyptium is an exception. The haemocoelic fluid is
the seat of election for the development and subsequent distribution
of the parasite in the adult tick.
Shortly after removal of the ticks from their hosts (cattle in this
instance) the early pre-flagellate forms may be found in the
haemocoelic fluid (PI. XXXVIII, figs. 1-5), which is a dark amber
colour, and is of about the consistency of human blood serum.
Microscopically it is seen to contain among other elements the large
vacuolated haemocoelic corpuscles. The nuclear and protoplasmic
portions of these cells, when coloured with Leishman's or Giemsa’s
5+9
solutions, show a tendency to stain, as do the nuclei, blepharoplasts
and protoplasm of human and animal parasites. This point is of
importance, as fragments of these cells might easily be mistaken
at first sight for parasites.
A few days after the ticks have been collected, the early pre¬
flagellate forms show evidences of division and increase in number
(figs. 5, 6, 8).
After about four to ten days, that is, a little time previous
to oviposition, early flagellate forms make their appearance
(figs, i i-l8). Rapid multiplication of the young flagellates now
takes place, with the formation of division rosettes (figs. 18 and 19),
and the growth to adult Crithidia (PI. XXXVIII, figs. 20-29;
PI. XXXIX, figs. 31-33) soon follows: —
The fully-grown Crithidia multiply by longitudinal division
(PI. XXXIX, figs. 30, 34-39) till the smallest drop of haemocoelic
fluid presents a felted mass of crithidial bodies and waving flagella.
Owing to the great numbers of parasites present, the haemocoelic
fluid is a milk-white colour. Towards the end of oviposition, and
just previous to the death of the tick, post-flagellate forms may be
found (figs. 40-44) in the haemocoelic fluid, although in some cases
all forms disappear from it. The post-flagellate forms also occur
in the genital tract, and are perhaps better termed ovarian forms:
they will be dealt with under the description of hereditary infection.
The intestinal diverticula and malpighian tubes have not been
found to harbour any of the parasites in the adult tick. Sometimes
early division forms (PI. XL, fig. 62) are found in the salivary
glands, but this seems to be the exception rather than the rule.
The parasites were found more commonly in female than in
male ticks.
MORPHOLOGY
The life cycle of Crithidia hyalommae in the adult tick may,
for the sake of description, be divided into four periods which,
however, merge into each other. These are the pre-flagellate, the
flagellate, the post-flagellate, and ovarian stages. The morphology
of the ovarian stage will be dealt with under a subsequent heading.
The morphology of the pre-flagellate, the flagellate and post-
flagellate stages may be described as follows: —
S5o
The Pre-flagellate Stage
The early pre-flagellate stages of C. hyalommae found in the
haemocoelic fluid are rounded or oval Leishmania-like bodies
(PL XXXVIII, figs. 1-5, 7, 8), which vary from 4 to 12/ti in length
and from 4 to 11 >u in breadth. The protoplasm of these forms is
of a finely reticulate nature, containing distinct vacuoles (fig. 4}
and a rounded nucleus which is situate near the periphery of the
parasite (figs. 1, 2). The blepharoplast is rounded or bow shaped
(figs. 1, 2), and acquires the characteristic dark tint of such
structures when coloured by the various stains. It may be placed
at the side of, or opposite to, the nucleus (figs. 3, 4).
These early forms^ soon show evidences of division (fig. 5), two
daughter parasites resulting, smaller in size and with more granular
protoplasm than their precursors. Plasmodial forms (fig. 6) may
result from rapid division. Increase in the size of the flagellate
bodies follows and the flagellum makes its appearance, arising in
the neighbourhood of the blepharoplast. The flagellum in its
outward growth carries with it a portion of the periplast which
forms the undulating membrane.
After the formation of the flagellum rapid division takes place,
rosettes being formed (figs. 18, 19), indeed nearly all stages from
rounded flagellates to nearly adult Crithidia may be seen in the
same specimen and in the same rosette (figs. 18, 19, 27). It may
also be noted that individual members of a division rosette grow at
unequal rates. The aflagellar end now grows outwards, at first
wide and blunted (figs. 25, 26, 29), but it soon narrows down and
tapers off into a fine point (figs. 27, 28).
The Flagellate Stage
The fully-grown flagellates present the crithidial type of body,
but are unusually long and slender (PI. XXXIX, figs. 28a-36).
The length of the body and flagellum varies from 26 /jl to 50/1, the
length of the flagellum alone being 12 $ p. The average breadth
of the nucleus varies from 2 n to 2*5 fi.
The aflagellar extremity tapers gradually to a fine point, while
the flagellate end of the body is continued for some distance along
the flagellum.
The flagellum, taking its origin from the vicinity of the
blepharoplast, runs along the free border of the undulating
membrane until the extremity of the body is reached. Here it
escapes from the periplast and projects freely for a considerable
distance. The flagellum is never connected with the blepharoplast.
A basal granule has been noted (fig. 31), but its presence is not
always evident, probably because of its close relation to the
blepharoplast. The undulating membrane is well marked, and in
living specimens its movements may be observed. In stained
preparations it may be seen thrown into folds.
The nucleus is roundish (figs. 28a-36) or oval, and generally
lies about the middle of the body, the whole breadth of which it
nearly occupies (figs. 28a-36). The structure of the nucleus varies.
At times the nucleus appears somewhat vesicular with a marked
karyosome (figs. 31-33); at other times the chromatin of the nucleus
is distributed in granules within the nuclear membrane (fig. 30).
When division is about to take place, the nuclear material is
generally more granular (fig. 30).
The blepharoplast of C. hyalommae is well defined in all
stained specimens, and colours deeply. Its position varies within
what might be termed the limits of crithidial definition (Patton,
1908). It may be closely applied to or a little distance from the
flagellar or aflagellar side of the nucleus (figs. 28a~33).
In all cases the flagellum arises in the close vicinity of the
blepharoplast, in whatever position the latter may be situated. Two
blepharoplasts may often be observed (figs. 30 and 35-38), but this
is really an early division phase. The blepharoplast is usually
rod-shaped (fig. 28a), and sometimes shows a diplosome appear¬
ance (fig. 34), probably indicative of approaching division.
Chromatoid granules may be present in the protoplasm. In
fresh specimens the protoplasm of the flagellar extremity of the
body is markedly granular.
The Post-Flagellate Stage
The post-flagellate forms seen in the haemocoelic fluid are
merely rounding-up parasites (PI. XXXIX, figs. 40-44). They are
similar to the ovarian forms which undergo the same process in
55 2
the walls of the female generative tract, and which will be
described when dealing with the hereditary infection.
Division
In the pre-flagellate stages division is initiated by the blepharo-
plast, which divides into two nearly equal masses. These masses
now move one on either side of the nucleus (PI. XXXVIII, fig. 5), or
form the base of a triangle with the nucleus at the apex (fig. 8).
The nucleus now divides into two, and when the binucleate system
is completed the protoplasm splits, two daughter forms resulting.
Large plasmodial forms may be produced in the haemocoelic fluid
before the flagella are formed (fig. 6).
In the early flagellate stages, the same phenomena are
observed; the blepharoplast is often bow-shaped before splitting
(fig. 11). The flagellum divides, commencing at its blepharoplastic
end (fig. 11) with the division of the blepharoplast and basal
granule. The blepharoplast and flagellum may have divided, and
protoplasmic cleavage may have commenced before the nucleus has
completed its division (fig. 15). The lashing movements of the
flagella at this stage may assist in this early protoplasmic division.
Fig. 15 shows this condition, with details of nuclear division: the
nucleus still shows the mantle fibres surrounding the centrodesmose
which connects the equatorial plates.
Division in the early flagellates is rapid, and division rosettes
are formed (figs. 18, 19). Two or more phases in division may
be observed in the same rosette (figs. 18, 19).
In the full-grown Crithidia longitudinal division takes place.
Before division the blepharoplast presents a diplosome appearance
(PI. XXXIX, fig. 34), and divides first, with the flagellum (fig. 35).
One portion of the blepharoplast and its future flagellum migrates
to the opposite side of the nucleus. The blepharoplasts may be
connected together by a thin chromatin line (fig. 36).
The nucleus, which is generally granular at this stage, divides
into two. The two flagellar ends separate, and the flagella, moving
rapidly, separate the remainder of the organism into two similar
adult Crithidia.
Division of the post-flagellate stages may occur in the haemo-
553
coelic fluid, but takes place to a marked extent in the ovarian
tissues (PI. XXXIX, figs. 51, 52), ovarian cells (PI. XL, figs.
55-57), and young ova (PI. XL, figs. 60, 61). Their division is
similar to the division of the pre-flagellate stages, and requires no
special description. Aggregation rosettes are formed (PI. XXXIX,
fig- 50 ).
Multiple division takes place in the deposited ova as in the early
stages in the haemocoelic fluid. These forms, the result of
multiple division, are multi-nucleated masses of protoplasm, and
have been termed ‘plasmodial forms’ (PI. XL, figs. 60, 61).
THE HEREDITARY INFECTION
The proofs of the occurrence of hereditary infection may be
studied by the examination of living specimens and stained
preparations. The process of events in the ovarian system is as
follows:
About the time of oviposition, and during this event, the adult
Crithidia pass by means of the haemocoelic fluid to the ovary and
oviducts.
A large number of the crithidial forms here begin the post-
flagellate ovarian stage of their existence in the walls (PI. XXXIX,
figs. 45-54) and cells (PI. XL, figs. 55-57) of these organs.
This post-flagellate or ovarian stage begins with the absorption
of the flagella and the gradual rounding off of the parasites
(PI. XXXIX, figs. 45, 46) to Leishmania-like forms (figs. 47-49).
A very large number of the parasites perform this evolution (fig.
50). A considerable number of the adult Crithidia, however,
pierce the walls of the oviducts, preferably high up where the oval
capsules are soft. Here they proceed to pierce the walls of the
ova (PI. XL, fig. 58), more than one parasite often entering the
same ovum (PI. XL, fig. 59). The aflagellar end is the fore¬
most end in entering. A boring motion may now be observed, the
flagellum describing a propeller-like movement, the whole organism
being screwed forward, the undulating membrane forming the
thread of the screw. Another motion has also been observed in the
ovarian forms. After rounding of the protoplasm of the flagellar
end, a circular motion has been noted which occurs previous to the
disappearance of the flagellum and formation of the resting
554
bodies. Once within the egg the rounding-up process described
above takes place, and the Leishmania-like stage results. Division
of these small forms may be found in the ova (PI. XL,
figs. 60, 61), which ultimately break up into small resting bodies.
A considerable number of the ova of infected ticks dry up and die.
Generally the later deposited ova contain the resting forms; the
ova deposited early in oviposition are usually non-infected.
SUMMARY
1. Crithidia hyalommae is a flagellate parasite occurring in
the haemocoelic fluid, salivary glands, ovary, oviducts and ova of
Hyalomma aegyptium , the common cattle tick of the Anglo-
Egyptian Sudan.
2. The parasite has four periods in its life-cycle: a pre¬
flagellate stage (PI. XXXVIII, figs. 1-8) passed chiefly in the
haemocoelic fluid, a flagellate stage in the haemocoelic fluid (Pis.
XXXVIII, XXXIX, figs. 9-39) and in the ovary and oviducts
(PI. XXXIX, fig. 50 and PI. XL, figs. 58, 59), a post-flagellate
stage (PI. XXX{X, figs. 40-44) in the haemocoelic fluid, and a post-
flagellate or ovarian stage in the ovary and oviducts (PI. XXXVIII,
XXXIX, figs. 45-54), ovarian cells (PI. XL, figs. 55-57), and ova
(PI. XL, figs. 60, 61).
3. The pre-flagellate stage is of short duration. The parasites
are small rounded or oval bodies from 4 n to 11 fi broad, and from
4M to 12 fi long. Tfceir protoplasm is reticulate or granular,
depending on division. They contain a nucleus and a rod-shaped
blepharoplast. Division of the very earliest forms takes place
(PI. XXXVIII, figs. 5-8).
4. The flagellate forms vary from 26 fi to 50 fi in length and
from 2 fi to 2*5 p in breadth.
The protoplasm is reticulate. The consistency of the nucleus
varies with division. The rod (PI. XXXIX, fig. 28a) or diplosomic
blepharoplast (fig. 34) is usually situated on the flagellar side of
the nucleus, but its position varies at times, depending on the phase
of division. The flagellum arises in the neighbourhood of the
blepharoplast, but not from it.
555
5. The undulating membrane and flagellum are both well
marked. The undulating membrane assists considerably in motion.
The flagellum forms the border of the undulating membrane,
and continues free from the protoplasmic body for a considerable
distance.
A basal granule has been noted (PI. XXXIX, fig. 31).
6. The post-flagellate and ovarian stages occur in the haemo-
coelic fluid, ovary and oviducts. A superfluity of the ovarian
forms seems to be present, a large number of them entering a
resting stage in the ovarian cells and tissues.
7. The ovarian stages of Crithidia hyalommae serve for the
hereditary transmission of the parasite. The flagellates pass by
means of the haemocoelic fluid to the ovaries and oviducts; here
they pierce the ova, the aflagellar end of the parasite being
anterior. Within the ova the flagellates round up and undergo
subsequent division with the production of ‘plasmodial 1 forms.
8. Multiplication of C. hyalommae by longitudinal division
takes place in all its stages.
9. Infection of Hyalommae aegyptium by Crithidia hyalommae
in the adult is purely a hereditary infection.
10. The flagellate is a strictly parasitic flagellate of the tick,
and is non-pathogenic to the tick host.
Acknowledgments
I wish to thank Dr. Balfour, the recent Director of these
Laboratories, and Dr. Chalmers, the present Director, for their
valuable assistance in both work and text; also Dx. Fantham,
whose experience in this line of research is well known, for his
valuable advice in the text and aid with the drawings. To
Dr. Bousfield and his able staff I am indebted for a plentiful supply
of working material.
Khartoum,
Septanber 27M, 1913.
556
REFERENCES
Flu, P. C. (1908). Ueber die Flagellaten im Darm von Melopbagus ovinus. Archiv fur
Protistenkunde, XII, pp. 147-153. One plate.
Mackinnon, D. L. (1909). Note on two New Flagellate Parasites in Fleas— Herpetomonai
ctcnopbtbalmi , n. sp., and Critbidia bystricbopsyUae , n. sp. Parasitology, II, pp. 288-296.
One plate.
O’Farrell, W. R. (1913). Preliminary note on a New Flagellate, Critbidia byalommae , sp.
nov., found in the tick Hyalomma aegyptium, Linnaeus, 1758. Journal of Tropical
Medicine and Hygiene, Vol. XVI, pp. 245*246.
Patton, W. S. (1908). The Life-Cycle of a species of Crithidia parasitic in the Intestinal
Tract of Gerris fossarum , Fabr. Archiv fur Protistenkunde, XII, pp. 131-146. One
plate.
- (1909). The Life-Cycle of a species of Crithidia parasitic in the Intestinal Tracts of
Tabanus bilarius and Tabanus , sp. Archiv fur Protistenkunde, XV, pp. 333-362.
One plate.
-(1909). A Critical Review of our Present Knowledge of the Haemoflagellates and
Allied Forms. Parasitology, I, pp. 91-143.
Patton, W. S., and Strickland, C. (1908). A Critical Review of the Relation of Blood¬
sucking Invertebrates to the Life-Cycles of the Trypanosomes of the Vertebrates, etc.
Parasitology, I, pp. 322-346.
Pfeiffer, E. (1905). Ueber trypanosomenahnliche Flagellaten im Darm von Melopbagus
ovinus. Zeitschrift fiir Hygiene und Infektionskrankheiten, I, pp. 324-29. One plate.
Porter, Annie (1909). The Morphology and Life-History of Critbidia gerridis , as found in
the British Water-Bug, Gerris paludum . Parasitology, II, pp. 348-366. One plate.
- (1910). The Structure and Life-History of Critbidia melopbagia (Flu), an Endo-
parasite of the sheep-ked, Melopbagus ovinus. Quarterly Journal of Microscopical
Science,Vol. LV, pp. 189-224. Two plates.
Sambon, L. W. (1903). Sleeping Sickness in the light of recent knowledge. Journal of Tropical
Medicine, Vol. V, p. 207, Col. 1, paragraph under Pig. 4.
Swingle, L. D. (1909). A Study on the Life-History of a Flagellate ( Critbidia melopbagi, n. sp.)
in the Alimentary Tract of the sheep-tick {Melopbagus ovinus). Journal of Infectious
Diseases, VI, pp. 98-121. Three plates.
55»
EXPLANATION OF PLATES
PLATE XXXVIII
All figures were outlined with the Abbe-Zeiss camera lucida,
after magnification with a 2 mm. apochromatic objective (Zeiss)
with No. 12 compensating ocular.
The magnification of the figures is approximately 2,000
diameters.
Figs. 1-4. Early pre-flagellate forms of Crithidia Jtycdommac
from the haemocoelic fluid.
Fig. 5. Young dividing form from haemocoelic fluid.
Fig. 6. Plasmodial form from haemocoelic fluid.
Figs. 7-8. Young forms from haemocoelic fluid.
Figs. 9-10. Rounded form showing flagellum.
Figs. 11-12. Early flagellates in haemocoelic fluid.
Figs. 13-16. Division phases in early flagellates.
Fig. 17. Early flagellate in haemocoelic fluid.
Figs. 18-19. Division rosettes, haemocoelic fluid.
Figs. 20-29. Development to full grown Crithidia, haemocoelic'
fluid.
560
Plate XXXIX
Fig. 28a. Fully grown Crithidia from haemocoelic fluid showing
rod-shaped blepharoplast.
Fig. 29a. Fully grown flagellate from haemocoelic fluid showing
blepharoplast on flagellar side of nucleus.
Figs. 31-33. Showing different positions of blepharoplast.
Figs. 34-36. Commencing division of blepharoplast. Fig. 34
shows diplosome blepharoplast. Fig. 36. Parasite
showing nuclear membrane and chromatic band from
haemocoelic fluid.
Figs. 30, 37, 38, 39. Stages in longitudinal division from
haemocoelic fluid.
Figs. 40-44. Post-flagellate stages in haemocoelic fluid.
Figs. 45-49. Post-flagellate or ovarian stage in the ovary and
oviducts.
Fig. 50. Shows aggregation rosette of ovarian stages in walls of
oviduct.
Figs. 51-52. Division stages of Leishmania-like forms of ovary.
Plate XL
Figs. 53-54. Division stages of ovarian forms of Crithidia
hyalommae.
Figs. 55-57. Resting and division stages in ovarian oells.
Fig. 58. Entry of adult Crithidia into wall of oviduct.
Fig. 59. Perforation of egg by adult Crithidia.
Figs. 60-61. Plasmodial forms in deposited ova.
Fig. 62. Early form (rare) in salivary gland.
Plate XL
Imid/x Trap. Mn/. (< fhrasifot., I o! I //
CRITHIDIA HYALOMMAE
\\!R.O'TorreU ilel.
S «3
TRYPANOSOMA VIVAX IN RABBITS
BY
B. BLACKLOCK
AND
WARRINGTON YORKE
(Front the Runcorn Research Laboratories of the Liverpool
School of Tropical Medicine)
{Received for publication 6 October , 1913)
It is generally accepted that T. vivax ( T . cazalboui ) is not
pathogenic for small laboratory animals. Thus Laveran and
Mesnil,* writing on T. cazalboui> state that the dog, monkey,
rabbit, guinea-pig, rat and mouse are refractory, and that this fact
constitutes one of the chief characters of the trypanosome. It is
true that Ziemannt records infection with T . vivax in dogs and rats,
but as short aflagellar trypanosomes were found by Ziemann in his
sheep and goats, it is highly probable that the animals from which
the dogs and rats were inoculated had a double infection of
T. vivax and T. congolense y and that the sub-inoculated animals
became infected with the latter parasite only. We might remark
in this connection that of late years it has been shown that double
infection of domestic animals and game with these trypanosomes
is by no means uncommon.
In previous communications* we have recorded an instance of
such a double infection in a horse naturally infected in the Gambia,
and described the manner in which the two parasites were isolated
one from the other.
This horse was the source of the strain of T. vivax with which
the present paper deals. As mentioned in a former paper, § the
vivax strain was separated fortuitously from the short aflagellar
parasite by inoculation of goats from the horse at different dates;
one of the sub-inoculated goats developed a pure culture of
* Laveran & Mesnil. Trypanosome* et Trypanosomiases. Paris, 1912, p # 542.
t Ziemann. Beitrag zur Trypanosomen Frage. Centralb. f. Bakt. Orig., 1905, Bd.
XXXVIII, p. 307, 429-662.
J Yorke Sc Blacklock. The trypanosomes found in two horses naturally infected in the
Gambia. Annals of Trop. Med. Sc Parasit., Vol. V, No. 3, p. 413.
§ Blacklock. The trypanosomes found in a horse naturally infected in the Gambia. A
double infection. Annals of Trop. Med. Sc Parasitology, 1912, Vol. VI, No. ib, p. 107.
564
T. vivax. From this time (June 14, 1911) up to the present the
strain has been preserved by passage through a series of thirty-nine
goats.
It is of interest to note that in spite of direct inoculation of the
strain from goat to goat during a period of nearly two and a half
years, no appreciable increase of virulence has been observed. Thus
the average duration of the disease in the first fourteen animals
used was thirty-one days, whilst that in the last fourteen was
twenty-nine days. The periods of incubation and duration of the
disease in this series of goats are given in Table I.
Table I. Showing a series of thirty-nine passages of T. vivax in goats during a period
of twenty-eight months.
Horse 1464
' Goat j '497 {;°
2 Goat 1559 |£J
3 Goat 1605
<9
Rabbit ibt>8
18
(5 mths.
Goat 1673 j^ 2 4 Goat 1584 a
r
Rabhit .717 {;j mths .
Rabbit 1747 neg.
1
Goat 1735 jig
Goat 1
8 5 8 {,"
Goat
1
I96sb|9,
I
Goat 2092 ^
I 6
1*9
5 Goat 1685 j*.
6 Goat 1733
7 Goat 1774
8 Goat 1801
I
7
16
9
27
*3
43
9 Goat 1838
10 Goat 1895
11 Goat ^1933
12 Goat 1965 a
13 Goat 1986
„ 1
14 Goat 1994
I
15 Goat 1994 A
Rabbit 1604
( 16
<57
Rabbit 1667 j
!
Rabbit 169! neg.
I
2 Rabbits 1736 neg.
I
Rabbit 1859 neg.
1 1
16 Goat 2058c Goat 2058B | Goat 20584A
M3
1 4 *
565
Table I —continued
17 Goat
18 Goat
19 Goat
20 Goat
21 Goat
22 Goat
23 Goat
2088 -''j
1 (37
r is
?“»* It,
is
,” 3; Us
r‘ U;
Goat 2165B [ ®
|
Goat 2194 [ i 4
1
Rabbits 2302 neg.
24 Goat 2288 j
1
[ 20
152
25 Goat 2326 |
1
in
1 3 *
1
Goat 2306 j® 2
1
Rabbits 2352 neg.
26 Goat 2359
f 9
1 29
27 Goat 2379 |
f 12
I24
28 Goat 2420 |
1 1 7
29 Goat 2448 j
1*
139
30 Goat 2496 |
!8
138
•31 Goat 2536 |
[6
s
*32 Goal 2545 |
Is
1
*33 Goat 2581 a •
js
1
•Goat 258 ib | ^
1 <
34 Goat 2602B -]
f 7
1 12
\
•35 Goat 2617 |
17
1
l
•Goat 2632B 15
*36 Goat 2632A \
[S
| |
*37 Goat 2642 1
j 5
| 1
38 Goat 2668 j
f 8
(no
Goat 2674 |^ s
39 Goat 2718 j
Ki -*4
00
* Goats marked thus used for treatment.
The upper figures in each bracket indicate incubation ; the lower, duration.
566
Attempts to infect rabbits, made in the earlier passages of the
strain through goats, either failed entirely or resulted in a temporary
infection in which the parasites were exceedingly scarce and
disappeared after a few days.* It was not possible to carry on
the strain to a second generation in rabbits.
Inoculation made from the thirty-eighth goat proved, however,
to be more successful, and four rabbits, injected intraperitoneally
with small amounts (05 to i c.c.) of the goat’s blood, all became
infected after incubation periods of five to eight days. The
infection in all was well marked, and parasites were fairly
numerous in the peripheral blood—up to twenty to a microscope
field. Two of the four died on the seventeenth and thirty-fourth
days, respectively, with numerous parasites in the peripheral blood.
The spleen was greatly enlarged and the animals were very
emaciated; the weights had decreased from 1220 to 970 grams and
from 1390 to 1000 grams. There has been some loss of weight in
the other animals, but after exhibiting parasites in the peripheral
blood for from three to eleven days they have since become
negative, and appear to have recovered.
There is, hence, evidence that the strain is, after thirty-eight
passages through goats, to a certain extent virulent for rabbits.
Three rabbits were inoculated intraperitoneally from the thirty-fifth
goat with about 30 c.c. of defibrinated blood. Trypanosomes were
found in the peripheral blood on the next day, and remained in
considerable numbers—as many as fifty to a microscope field—
until the time of death, which occurred on the fifth, ninth and
fourteenth days, respectively. All the animals presented marked
signs of infection, the spleens were greatly enlarged, and the
animals had lost a considerable amount of weight.
From one of these (Rabbit 2627B), two others were inoculated
intraperitoneally with 1 c.c. of citrated blood (containing about
0*25 c.c. of blood). Trypanosomes were found in the peripheral
blood of one of the two sub-inoculated animals on the sixth, and
it died on the fifteenth day after inoculation. The strain has since
been carried on in rabbits over a period of more than three months,
until the eighth generation, as shown in Table 2.
• Blacklock. A note on T. vivax in rabbits and white rats. Annals of Trop. Med. &
Parasitology, 1912, Vol. V, No. 4, p. 537.
Table II. Showing a series of eight passages of T. vivax in Rabbits during a period of three months.
Goat 2617
568
From a rabbit of the sixth generation a goat was inoculated
subcutaneously with i c.c. of citrated blood, and trypanosomes
appeared in its blood on the ninth day.
Although, as the table indicates, the strain has not yet become
invariably pathogenic for rabbits, and a considerable number
appear to have recovered, nevertheless most of the animals have
developed a definite infection, and in five the disease has run an
exceedingly acute course, the animals dying in from five to twenty
days with numerous parasites in the peripheral blood.
We have examined the strain after passage through six
generations of rabbits, with a view to ascertaining whether it has
undergone morphological changes. In this connection we might
mention that as a general rule the trypanosomes in rabbits do not
exhibit that rapidity of movement which is characteristic of them
in goat’s blood. In stained*preparations the trypanosomes appeared
to be identical with those seen in goat’s blood.
We intend, if possible, to carry on this strain in rabbits in the
hope of rendering it acutely pathogenic for other small laboratory
animals.
5*59
THE PATHOGENICITY OF NOSEMA
APIS TO INSECTS OTHER THAN
HIVE BEES
BY
H. B. FANTHAM, D.Sc. (Lond.), B.A. (Cantab.)
(LECTURER ON PARASITOLOGY, LIVERPOOL SCHOOL OF TROPICAL MEDICINE)
AND
ANNIE PORTER, D.Sc. (Lond.), F.L.S.
(quick LABORATORY, CAMBRIDGE)
(Received for publication 31 October , 1913)
CONTENTS
Introduction
PAGE
. 569
Experimental Infection of Various Insects ...
. 57°
A. Hymenoptera
(i) Humble Bees ... .
...
570
(ii) Mason Bees .
. 570
(iii) Wasps.
.
. 571
B. Lepidoftera
(i) Cabbage White Butterflies
...
. 572
(ii) Peacock Butterflies .
..
. 572
(iii) Cinnabar Moths .
...
. 573
(iv) Gooseberry Moths .
.
. 574
C. Diptera
(i) Blow Flies .
.
. 575
(ii) Crane Flies .
...
. 576
(iii) Sheep Keds .
.
. 576
Summary and Conclusions .
..
578
References ... ... ... .
.
. 579
I. INTRODUCTION
Nosema apis is now well known as the minute, microsporidian
parasite that causes a malady of hive bees, popularly known as
‘Isle of Wight* disease, though it should be noted that several
diseases of bees are often being confused under this general name.
The organism, N. apis , is transferred from bee to bee by means of
resistant spores in the dejecta of the infected host, which spores are
taken up by other bees, either with food or drink, or during the
various processes consequent on the communal life of the bees.
The Nosema spores absorbed by the bee liberate amoeboid forms
termed planonts, which enter the epithelial cells of the gut and
57o
develop rapidly, particularly in the chyle stomach of the bee
where they undergo rapid, asexual multiplication, forming meronts.
The functional derangements resulting therefrom are quite sufficient
to bring about the death of the host in many cases. Should the
host react successfully on the parasites, spore formation ensues, and
the Nosema leave the host as highly resistant spores that are well
adapted for extra-corporeal life, and that serve to perpetuate the
species should they be able to reach a new host.
Some experiments have been carried out with insects other than
Apis mellifica , and tend to show that the pathogenicity of Nosema
apis is far from restricted to the hive bee. Many of the experi¬
ments were suggested by observation of the habits of various insects
present when examination of bees was in progress. Although the
numbers of the insects used were not as large as we could have
wished, the results are of interest and importance.
II. EXPERIMENTAL INFECTION OF VARIOUS INSECTS
Experimental infections of members of the Hymenoptera,
Lepidoptera and Diptera have been made.
A. HYMENOPTERA .
The experimental insects used were humble bees, mason bees
and wasps.
(i) HUMBLE Bees (Bombus terrestris y B, lapidarius , B. hortorum ,
B. venustus , B. latreillelus). It has been shown previously by
Dr. Graham Smith and ourselves that a Nosema is a natural parasite
of humble bees (Bombus spp.). It seems to be a different species
of Nosema from that so destructive to hive bees, and chiefly
parasitises the malpighian tubules of the humble bees.
But humble bees belonging to apparently clean stocks, when
kept in captivity and provided with food contaminated with excre¬
ment of hive bees containing Nosema apis spores, both can, and do,
become infected with the parasite, and die from the effects thereof.
Both meronts and spores can be found in the walls of the gut,
while the malpighian tubules, as is usual with infections of Nosema
apis , remain uninfected. Humble bees used as controls and
supplied with pure food showed no Nosema.
(ii) MASON Bees. The colonies of mason bees used were
\
n
57 1
brought from abroad and were lodged in a piece of old wall.
Some of the bees were isolated to act as controls. The rest were
kept in a wall, screened off to prevent exit, and were supplied with
honey and pollen contaminated with spores of Nosema apis from
dead bees. The abundant food supply proved attractive and the
bees fed somewhat greedily. On the fourth day after the first
supply of contaminated food, it was noted that fewer bees flew
out, and a few dead ones were found. On dissection, the
alimentary tract of these bees was shown to be parasitised with
meronts of N. apis. The dwindling of the bees continued, and
some were found unable to fly. N. apis in the form of meronts
and young spores was demonstrable on examination. Ultimately
the two colonies supplied with N. apis died out, and the brood also
was found dead. A few larvae showed Nosema spores, but the
brood had been too long dead when recovered for the determina¬
tion of young stages of the parasite. When both colonies were
extinct, the controls that had been supplied with pure honey were
examined. Seven-eighths of them had survived, and when these
were dissected no parasites were found. The controls that died
were also dissected, but as their food canals were empty, and their
fat bodies greatly reduced, their deaths were ascribed to voluntary
starvation.
(iii) WASPS (Vespa germanica ). Two years ago we reported
that a colony of wasps had been exterminated by introducing bees
dead of Nosema into their nest. Since then, the experiment has
been repeated, with similar results, and examination of recently
dead wasps has shown that multiplicative stages (meronts) of
Nosema apis were present in numbers in the alimentary tracts of
the wasps.
Again, wasps collect dead and dying bees and carry them
away as food for their larvae. Several cases are known to us in
which the wasp broods have died out entirely as the result of a
liberal diet of bees dead of Nosema. As before, the parasite
underwent development in the body of its host, and in the
majority of the wasps, adults and larvae alike, the host was killed
before the life-cycle of the parasite was completed by spore
formation. The hypothesis that the newer a parasite is to its host,
the greater is its virulence, thus receives further support.
572
B. LEP 1 D 0 PTERA.
Certain common Lepidoptera were used for experimental
purposes, controls being kept in each case.
(i) Cabbage White Butterflies (Pieris brassicae). Both larvae
and imagines were used for experiment, as both could possibly
acquire Nosema apis naturally with their food.
(a) An adult, » was fed on the day it emerged from the pupa
on syrup made with castor sugar and contaminated with spores
from the gut of a bee. The butterfly was fed again three days
later, and died on the fifth day after the first feed. Nosema
spores and young stages of the parasite were found in the gut.
The control died on the sixth day, and no parasites occurred in it.
( b ) An adult, 9 > was fed on the second day after escape from
the pupa on sugar syrup contaminated with spores. It refused to
feed on the first day. It lived four days after emergence. A few
Nosema meronts were found in the walls of the gut after death. A
control lived six days.
( c ) Larvae. Cabbage plants near a badly diseased hive were
found spattered with bee excrement. Dead larvae of cabbage
white butterflies also were found. Several experiments were then
made, of which the one cited was typical. Eight larvae were
collected from clean cabbage plants. Four were fed on a
cabbage leaf smeared with honey in which an infected bee’s gut had
been emulsified, the remaining four on ordinary clean cabbage
leaves.
One experimental larva died after two days with Nosema
infection in its gut. Two, also containing Nosema, died on the
following day. The remaining one pupated on the third day, but
the imago never emerged. Of the four control larvae, all pupated
and produced imagines in due course.
(ii) Peacock Butterflies {Vanessa io ). On two occasions
when bees were under examination for Nosema apis , peacock
butterflies came into the laboratory, and after the usual aimless
flutterings, settled on the viscera of dead bees and proceeded to
suck honey from the contents. Both were captured and were
examined after death. A second butterfly was captured on each
occasion to act as control.
{a) Adult, S- It lived two days after capture. When
573
dissected, no stages of Nosema apis were found in the gut walls,
and only a very few Nosema spores—probably those ingested with
the food—were present.
(b) Adult, 9 - This was a rather large female. Soon after
feeding, oviposit ion began, and the insect died at its termination.
No Nosema was found in it.
The extremely small number of experiments prevents any
definite conclusion being reached as to the pathogenicity of the
protozoon to the insect. All that can be said is that a negative
result was obtained in the two cases investigated.
(iii) CINNABAR Moths (Callimorpka jacobeae ). Larvae of the
cinnabar moths were used for experiment as they were observed on
several occasions feeding on groundsel contaminated with bee
excrement. The subjects experimented on were obtained from a
locality where bee-keeping was not practised, and where the larvae
seemed remarkably healthy.
Fifteen young caterpillars of the cinnabar moth were placed on a
plant of groundsel growing in a flowerpot and covered by a bell
jar to prevent escapes. The groundsel was watered with an
emulsion of dead bees containing Nosema spores, filtered through
coarse muslin that readily allowed the Nosema spores to pass
through, but retarded the passage of the chitinous portions of the
bees. The groundsel foliage was watered daily. The larvae fed
on the groundsel in a quite normal way. Control larvae were kept
under similar conditions but their groundsel was watered with
ordinary tap water. These control larvae never showed any form
of ’Nosema apis . The results of the experiment were: —
Three caterpillars, fed on infected groundsel, died two days
after the first infective feed. Young stages of Nosema apis were
present in their mid guts. The hind gut was not affected. The
parasites were in the multiplicative phase, meront formation being
in progress.
Seven larvae died on the fourth day. Of these, one contained
young spores of Nosema apis f two had a very heavy infection of
young meronts in an actively dividing condition, and the remaining
four showed a few young stages of the parasite. The condition
of the guts of the larvae resembled that seen in bees, and the
varying degrees of infection among the larvae are parallel to what
574
we have so often found when examining a series of bees from one
colony.
One larva died on the fifth day after the first feed, and con¬
tained mcronts of Nosema apis. On the same day one pupated, and
ultimately a deformed male imago issued from it. This insect
lived three hours only. It was dissected as soon as possible after
death, but no stages of Nosema apis could be recognised with
certainty within it.
The remaining larvae died during the night and were too
decomposed when examined to permit the detection of the planonts
or meronts of Nosema apis , had they been present. No spores
were found.
The life cycle of Nosema apis as seen in the larvae of the
cinnabar moth was the same as that found in the hive bee. The
spores gave rise to planonts, which became meronts in the epithelial
lining of the alimentary tract, and these, in turn, produced spores.
(iv) GOOSEBERRY Moths (Abraxas grossulariata). Observa¬
tions on a garden in a district heavily infected with apian
microsporidiosis showed that, in one case, the foliage of gooseberry
bushes near infected hives was spattered with the excrement of the
bees, and the dejecta on the leaves contained spores of Nosema
apis . Beneath the bushes a few dead larvae of the gooseberry
moth were found, and as the body of one of these contained Nosema
spores, experiments with larvae obtained from an uninfected district
were commenced. Twelve larvae were divided into two sets of
six. Both were fed on gooseberry twigs, the twigs being moistened
daily with equal quantities of water, and of water containing
Nosema spores, respectively. The results may be summarised
thus: —
(a) Larvae supplied with food contaminated by Nosema spores .
On the fifth day after the first infective feed, one larva died.
Nosema spores were found in its intestine, toegther with meronts.
These spores were fed to bees and reproduced the disease.
On the seventh day two more larvae were found dead. Thirty-
six hours had elapsed since they were last seen alive. Decay had
been rapid, and it was impossible to identify young stages of
Nosema apis , and no spores were found. Thus it is uncertain
whether these two larvae became infected.
575
One larva died on the eighth day. It contained both spores and
meronts of Nosema apis.
Two larvae pupated on the tenth day. The pupae were small
compared with those from the control larvae. One pupa was
dissected on the twelfth day, and a few meronts were found in the
body. The second pupa did not produce an imago. Whether
this was the effect of the action of the Nosema cannot be stated
with certainty.
( b ) Larvae feeding on uncontaminated food . Two of the
larvae were dissected. Neither showed any trace of Nosema apis.
The remaining four pupated. Two pupae were dissected and
examined. Nosema apis was absent, nor was any other parasite
found in them. Of the remaining two pupae, one produced a
beautifully shaped and marked female, but the second did not
develop.
Morphologically, the Nosema present in the larvae of the
gooseberry moth differed in no wise from the parasite as seen in
bees, and its identity was established by feeding bees with the
spores obtained from the caterpillars and thereby reproducing the
disease.
C. DIPT ERA.
The Diptera used for experiment were blow flies, crane flies and
sheep keds.
(i) BLOW Flies ( Calliphora erythrocephala ). During the
examinations of bees for Nosema apis , it was noticed that blow¬
flies settled on the viscera of the bees and fed upon them. The
flies were also often seen sucking up the sweet excrement voided on
the alighting board and sides of the hive; they subsequently
showed Nosema infection.
A number of pupae of Calliphora erythrocephala were dug up
in a garden on March 26th, 1912. The adult flies began to emerge
on March 28th, but could not be induced to feed %n that day. On
March 29th, the flies sucked a piece of meat with some Nosema
spores on it. The following day they were supplied with moistened
sugar contaminated with Nosema spores. They refused again to
feed, but sat on the sugar most of the time.
From April 1st to April 9th, the blow flies hatched out at the
576
rate of one or two a day, and were formed into experimental and
control sets. Special cases are now cited: —
Blow Fly i emerged from the pupa. It was provided with
contaminated sugar and sucked it readily. It died nine days after
emergence, and on dissection showed some young stages and a very
few spores of Nosema apis in its gut. Infection of the Malpighian
tubules also occurred.
Blow Fly 2 was smaller than Fly I, and emerged a day later
than it. It also lived nine days, while its control lived ten days.
At death it contained a number of Nosema spores.
Blow Flies 3, 4, 5 and 6 lived from 7 to 10 days after emergence.
All were fed on infected candy. Blow Fly 3 became infected,
showing a few spores at death. Blow Fly 4 showed no form of
Nosema apis. Blow Flies 5 and 6 contained a very few spores of
N. apis . Blow Fly 7 showed no Nosema apis , but contained
what is probably a new species of Nosema. Blow Fly 8 contained
many young stages of Nosema apis, together with a fair number
of spores. Blow Flies 9 and 10 contained no parasites.
No control blow fly was found to harbour microsporidia.
From the above experiments it can be inferred that a certain
number of Calliphora are attacked by Nosema apis f and the latter
can prove fatal to them if they are ingested by the insect with its
food.
(ii) CRANE Flies (Tipula olcracea). Two crane flies were
noticed sucking the viscera of bees prepared for microscopical
examination. The crane flies were captured, but refused to feed
in captivity and died two days after capture. When dissected, a
few young stages of Nosema apis were found. Crane flies caught
in the open and used as controls showed no trace of microsporidian
in fection.
(iii) SHEEP Keds (Melopkagus ovinus'). All the insects
previously mentioned were able to acquire spores of Nosema apis
naturally by means of their food, though the number of cases
occurring in the open may not be very considerable. During the
course of this investigation, a number of healthy sheep keds,
Melophagus ovinus , from a prize Southdown flock, were sent to us.
Though it was not very probable that these insects could obtain
Nosema spores in nature, it was considered that it might be of
577
interest to see what effect, if any, was produced on them by Nosema
apis. In order to ensure the ingestion of Nosema spores by the
keds, spores from the guts of bees were smeared on to a limited
area of the forearm of one of the writers and six keds were fed on
this small area. When wounds were made by the bites, additional
small drops of infected excrement were placed on them, and through
the layer of spore-containing material the keds had to force their
proboscides to get blood. Some of the keds after sampling the
sweet excrement slowly sucked it up before taking much blood.
All six fed well, the process taking nearly an hour in every case.
Six control keds were fed on a carefully disinfected area of the
other arm of the experimenter. The next day the treated keds
were alive but not active, as it was a very cold day. They were
fed again in the same way, all feeding well, but three being
particularly greedy. A third feed was given on the next day,
when the three weaker feeders of the previous day also fed slightly.
Examination of the excrement of these three showed that they
contained a fungus, as reported by one of us in 1910.* The other
three showed no fungus in their faeces. At noon on the fourth day
after the first feed, the three keds containing fungus were dead.
Microscopical investigation showed that they contained large
quantities of fungus in their Malpighian tubules and young stages
of Nosema apis in their mid guts. Soon after mid-day, the
remaining keds were noticed to be much more feeble, and between
3 p.m. and 9 p.m. that day all three died. Examination of fresh
preparations of them showed the presence of meronts and empty
sporocysts of Nosema apis in the gut contents. Stained prepara¬
tions were also made and were confirmatory of the fresh
preparations; they also contained young spores. When the control
keds were dissected, no form of Nosema apis was observed in any
of them. From this experiment, it is shown that Hippoboscid
flies, Melophagus ovinus , became infected when supplied with
spores of Nosema apis , which underwent developmental changes in
their bodies.
The Glossinae or tsetse flies resemble the Hippoboscidae in their
•Porter, A. (1910). The Structure and Life-history of Critbidia melopbagia (Flu), an
Endoparasite of the Sheep Ked, Melopbagus ovinus. Quart. Joum. Microsc. Sci., LV, pp. 189-
224. Two plates.
578
mode of reproduction and blood-sucking habit. As is well known,
the Glossinae are the flies that transmit sleeping sickness or
trypanosomiasis of men and animals. We would suggest that a
search should be made in the various Glossinae by competent
observers, who are well versed by practical experience in the
structure and life history of Microsporidia, for parasites allied to
the Nosema so destructive to bees, and pathogenic also to mason
bees, wasps, and the various Lepidoptera and Diptera cited above.
Should such a pathogenic Microsporidian be found as a hyper¬
parasite in Glossinae, it would be a forward step in the solving
of the problem of sleeping sickness.
III. SUMMARY AND CONCLUSIONS
1. Nosema apis has been proved pathogenic to Hymenoptera
other than bees. It can multiply in the food canals of humble bees,
mason bees and wasps, and can bring about the deaths of the hosts.
2. Contamination of plants with infected excrement occurs in
the neighbourhood of badly infected hives. Such contaminated
food is pathogenic to the larvae of cabbage white butterflies,
cinnabar moths and gooseberry moths, in which Nosema apis
produces destruction of the tissue of the food canal in the same
way as in bees. Both imagines and larvae of these insects became
infected with microsporidiosis when supplied with food con¬
taminated with Nosema spores.
3. Calliphora erythrocephala. The blow fly becomes infected
naturally by ingesting Nosema spores contained in the sweet
excrement of bees. This infection has been repeated experimentally.
Crane flies may also become infected.
4. A member of the Hippoboscidae, Melophagus ovinus , has
been infected successfully with Nosema apis , which is pathogenic
to the sheep ked. It is suggested that research be made by
competent observers among the Glossinae for Microsporidian
parasites allied to the Nosema of bees, and, possibly, equally
pathogenic to the tse-tse flies that may harbour them.
579
REFERENCES
Favtham, H. B., and Poktei, A. (1912). Microsporidiosis, a protozoal disease of bees due to
Nosema apis , and popularly known as * Isle of Wight * disease. Annals Trop. Med. and
Parasitol., VI, pp. 145-161.
-(1912). The Morphology and Life-history of Nosema apis , and the significance of
its various stages in the so-called 1 Isle of Wight * disease in bees (Microsporidiosis).
Annals Trop. Med. and Parasitol., VI, pp. 163-195. Three plates.
-(1912). The Dissemination of Nosema apis. Annals Trop. Med. and Parasitol.,
VI, pp. I97-H4-
Report on the 1 Isle of Wight * Bee Disease (Microsporidiosis). Supplement No. 8, Journ.
Bd. Agric., May, 1912.
Further Report on the * Isle of Wight 1 Bee Disease (Microsporidiosis). Supplement No. 10,
Journ. Bd. Agric., July, 1913.
58 i
ON CERTAIN MOSQUITOS OF THE
GENERA BANKSINELLA , Theobald,
AND TAENIORHTNCHUS , Arribalzaga
BY
HENRY F. CARTER, F.E.S.
(LECTURER IN ENTOMOLOGY, LIVERPOOL SCHOOL OF TROPICAL MEDICINE)
(Received for publication 28 'November , 1913)
During the past few weeks I have made a critical examination
of the male genital armatures of numerous species of African
mosquitos and, in certain instances, somewhat interesting results
have been obtained from their subsequent study. These studies
are more especially connected with the affinities of certain obscure
species and in reference also to the synonymy adopted by other
students of this group of blood-sucking insects.
Before dealing with the specific characters, it would perhaps not
be out of place in this paper, if I draw attention to the fact that
the structure of the male genitalia does not support the separation
of the pale coloured African species of Chrysoconops , so defined
by Theobald, from the genus Taeniorhynchus. The armatures of
typical examples of the latter genus compared with those of the
African Chrysoconops are so essentially similar that I consider them
to be congeneric.
BANKSINELLA palpale (Newstead).
Neomelanoconion palpale Newstead. Ann. Trop. Med. and
Parasit ., I, p. 31 (1907):
Banksinella luteolateralis , Edwards ( nec Theob.). Bull. Ent Res. t
III, p. 6 (1912).
This species was originally described by Newstead (/.c.) from
a single male collected by Drs. Dutton and Todd at Boma, Congo
Free State. Unfortunately, however, the specimen was somewhat
rubbed and therefore certain of the more important characters
rendered obscure. Last year the type was submitted to
582
Mr. Edwards, of the British Museum, for examination, who
subsequently placed the species as a synonym of B. luteolateralis,
Theob. Some time ago a perfect male example* of an apparently
new species was received from Broomassie, Ashanti. The armature
of this specimen was prepared for microscopical examination, and,
on comparison with a similar preparation of the type of N. pal pale ,
was at once seen to be the same. Newstead’s species is therefore
a valid one, and must be raised to specific rank.
Male genitalia (fig. i).t
Fio. ib. Inner literal
* * c view of right ciaip
* filament (cf.) of
Fio. i a. Male genitalia of Banksinella palpal*, Newst. x 160; B. palpalt, Newst. x
• = lanceolate spinet 240
Side pieces relatively large, broad posteriorly, narrow and rod¬
like distally, the internal edges being somewhat strongly curved, as
shown in the figure. The latter bear numerous fine hairs in the
central region, extending from each side towards the middle line,
* Mr. Edwards has kindly examined this specimen for me.
t In the designation of the parts I have followed Messrs. Howard, Dyar and Knab
and the main lettering in all the figures is as follows :—
s.p. = side piece; c.f. = clasp filament; u = unci; h = harpe ; b.l. = haul
lobe of side piece.
5»3
and several long, markedly curved ones on the apical lobe-like
projection; between these groups of hairs are situated six or seven
lanceolate spines (fig. I s), but the number of these appears to vary,
since in the specimen from Ashanti only three are visible. Clasp
filament comparatively short and stout, expanding slightly on the
inner side near the middle, tapering off towards the apex, and bearing
numerous curved hairs and a large conspicuous, slightly curved tooth¬
like projection in this region. The inner lateral surface of this organ
(fig. i b) is considerably broader, with a blunt and rounded apex;
the hairs and tooth previously mentioned, arise from the apical
portion of this surface and extend towards the middle line. Basal
lobes small, each bearing two distinct spines or teeth, and a few
Fig. ia. Left side-piece and appendages
of Banksinella luteolatcralis , Theob. x 160 j
s *2 lanceolate spines.
Fig. 2 b. Left clasp filament
enlarged ( X 240)
delicate hairs. Harpes well developed, the apical extremity
pointed, forming a single tooth. Harpagones absent. Unci of
complex structure bearing three pairs of comparatively large,
inwardly and ventrally directed, teeth on the apical ventral margin;
these teeth increase proportionately in size, the lowermost pair being
the smallest.
The genital armature of B. luteolateralis (fig. 2) is very similar
to that of the previous species as regards the structure of the basal
parts, and general form of the side pieces. The clasp filaments
584
(fig. 2 b ) provide, perhaps, the most distinctive character. These
are of a somewhat different shape, and are completely devoid of
hairs at the apical extremity; the tooth, also, is distinctly larger.
The internal edges of the side pieces bear similar arrangements of
hairs and spines as in B. palpale , but the hairs of the central region
are less numerous, and the lanceolate spines are smaller and about
fifteen in number.
As previously mentioned, the type of Banksinella palpale was
in a somewhat damaged condition when described, and I therefore
thought it desirable to add a few notes on the specific characters
of the male.
Head : The narrow, curved scales occupy a narrower median
area than in B. luteolateralls , Theob., and the pale coloured region
is formed to a considerable extent by cream coloured flat scales.
Fig. 3. (a) Male palpus of Banksinella palpal #, Newst. ; (A) Male palpus of BankstnelU
luteolateralls , Theob. ; x 36.
Palpi with the second or terminal segment relatively much
shorter than in B. luteolateralis ; the basal segment is nearly three
and one-third times the length of the apical segment, whereas in
B. luteolateralis it is approximately two and one-third times as long.
Proboscis sometimes showing an ill-defined yellowish band.
Thorax : Clothed with golden, narrow-curved scales laterally,
dark curved scales and a few scattered golden ones in the central
region—similar to the ornamentation usually met with in this genus.
Abdomen : First two segments dark, unbanded; third and
fourth with basal lateral cream coloured spots, especially noticeable
on the latter; fifth, sixth and seventh segments* with well-marked
yellowish-white basal bands. In the male of B. luteolateralis the
first and second segments usually possess a few pale scales on the
median area, the others being adorned with basal pale bands.
Wings: Very similar to those of B. luteolateralis .
Legs : Brown, the femora and tibiae being pale on the ventral
surface; tibiae with apical knee-spots, well marked in the hind pair
of legs.
Taeniorhynchus maculipennis (Theobald).
Chrysoconops maculipennis , Theobald. Novae Culicidae , Part I,
p. 27, April, 1911.
This species, described by Theobald from Uganda, has recently
been placed as a synonym of Taeniorhynchus ( Chrysoconops )
annettii , # Theob., by Mr. Edwards. Through the kindness of
Mr. H. H. King, of the Wellcome Laboratories, a male example has
been received by the School, and an examination of the genitalia
at once proves the species to be a valid one. Mr. Edwards writes
that he has now examined preparations of the armature and is in
a position to confirm the above statement.
Male genitalia (fig. 4.)
Side pieces large, gradually tapering to a broadly rounded
apex. Clasp filaments long and relatively narrow, each bearing
two short closely appressed teeth at the tip. The apical third bears
four delicate hairs, and is deeply constricted at its base, on the lower
side. Basal lobes large, each with a pair of dark rod-like
appendages; one of the latter, in each case, is a very conspicuous,
stout, elongated structure, the other much narrower and slightly
shorter. The larger rod-like process is evidently composed of three
or four fused spines or modified hairs, the smaller rod of one only.
• The last few segments are necessarily wanting, as they were removed for microscopical
examination.
586
Harpes well marked, with five comparatively large teeth at the
extremities. Harpagones absent. Unci as depicted (fig. 4»),
bearing numerous minute teeth on the apical, dorsal and ventral
edges.
u
H.
B.L
R
SP
CF
Fio. 4 Male genitalia of Taenicrbyncbus maculipennu , Theob. ; x 160.
Genital armature of T. annettii (fig. 5).
The main point of difference between this and the previous
species is in the structure of the clasp filaments. These are of
very peculiar form, and an idea of their shape can best be gained
from the figures. The apical portion is broad and very deeply
cleft on the ventral surface; it bears a few delicate hairs and three
teeth, closely pressed together, at the tip.
The basal parts closely resemble those of T. maculipennu ,
although the harpes appear to be more elongated (vide fig.) This,
however, may be due to displacement of the parts in mounting,
588
but, owing to lack of material, no definite statement can be made.
The identification of these two species by other means is a matter
of no little difficulty, and, unfortunately, the material at my
disposal is insufficient to enable me to give any constant and well-
defined characters for distinguishing purposes.
Taeniorhynchus METALLICUS (Theobald).
Culcx metallicus , Theobald. Mon. Cul. , II, p. 63 (1901).
Banksinella metallicus (Theobald). Mon . Cul., V, p. 408 (1910).
Taeniorhynchus violaceus, Theobald. Third Report Wellcome
Labs., p. 262 (1908).
The examination of preparations of the male armature of
authenticated specimens of T . metallicus from various parts of
Africa, and of T. violaceus from the Sudan (Mr. H. H. King) show
that the above synonymy', previously proposed by Mr. Edwards, is
correct.
Male genitalia (fig. 6).
Fig. 6. Male genitalia of Taentorbyncbus metallicus , Thcob.; x 160.
\
5 8 9
Side pieces sub-cylindrical, adorned with the usual stout and
slender hairs, and bearing basal lobes with their appendages, which
greatly resemble those of the two above-mentioned species. Clasp
filament very characteristic, the apical two-thirds being expanded
for part of its length, then tapering gradually to the tip, on which
is situated a very short, stout tooth; the lower side of the apical
half is sharply re-curved, forming a distinct ridge. Harpes
apparently provided with three teeth. Harpagones absent. Unci
with a series of eight or nine small teeth on the ventral basal edges,
and four larger teeth at the apex.
REFERENCES
Edwards, F. W. (1911). The African species of Culcx and allied genera. Bull. Ent.
Res., II, pp. 241-268
- (1912)- A synopsis of the species of African Culicidae, other than Anopheles. Bull.
Ent. Res., Ill, pp. 1-53.
- (1913)- Further notes on African Culicidae. Bull. Ent. Res., pp. 47-59.
Howard, L. O., Dyar, H. G., and Knab, F. (1912). The mosquitoes of North and Central
America and the West Indies, I, pp. 69-71.
59 1
NEW CULICIDAE FROM THE SUDAN
BY
FRED. V. THEOBALD, M.A., F.E.S., Hon. F.R.H.S.
(.Received for publication 24 N'ovember , 1913)
Amongst a large collection of Culicidae sent me by Mr. Harold
King, from the Sudan, the following undescribed species have so
far been found.
A considerable number of Uranolaenias and others have as yet
only been partially examined, and will be reported upon later.
The types of these new species are in the collection of the Liverpool
School of Tropical Medicine.
Mucidus NIGERRIMUS, nov. sp.
< 5 . Head : Brown, covered with mealy grey scales, a narrow
white border around the large dark eyes, a tuft of long grey curled
scales and long, thin, straight ones projecting forwards between
them. Antennae brown, with deep brpwn plume hairs; basal
segment bright brown, with small flat white scales. Palpi deep
brown, with black scales irregularly disposed and scattered white
scales, especially at the junctions of the last two segments, forming
distinct bands; plume hairs deep brown, apical segment slightly
longer than the penultimate; longer than the proboscis by about
two-thirds of the last segment. Proboscis with scattered dark and
light scales, as in the palpi, a distinct joint about the middle, the
apical half being thinner than the basal.
Thorax : Brown, clothed with irregular scattered grey scales,
most dense in the median area and around the space before the
scutellum and across the thorax between the wings. Scutellum very
densely clothed with loose grey scales; the scantier scaled areas
have long, thin, narrow-curved, pale scales, not typical Mucidus
ones, which also form a dense scaled median area in front.
Pleurae very deep brown, with patches of flat white scales.
Metanotum rich brown.
592
Abdomen : Deep brown, clothed with alternate patches of
black and white scales, forming marked black and white out¬
standing lateral patches; hairs dense, pale golden; the segments
dorsally, with flat, loose white scales at the base of the segments,
in the middle, with mixed dark and creamy ones apically and at
the sides more evenly disposed; the apical segments more white
scaled, the last but one with two black spots. Basal lobes of
genitalia, with black scales; claspers long, curved, dark.
Legs : Fore femora thin, with dark and pale scales, the former
predominate, apex white; fore tibiae with a dull white basal band,
followed by a black area with outstanding scales and then a snow-
white area with outstanding scales; tarsi thin, yellow, slightly
darkened apically; mid legs with the femora with dense out¬
standing scales, black predominating, with three narrow white
bands and white apex; tibiae with white basal and apical bands,
black in middle, with a narrow median white band, very shaggy;
tarsi thin and yellow; hind legs with femora and tibiae as in the
mid, but not quite so shaggy or dark; first tarsal black scaled, with
narrow basal white band, yellowish at apex, very shaggy; remaining
tarsals white basally, yellow apically, scales more or less out¬
standing; the white predominate on the last three segments; hind
ungues equal and unisecrate, fore and mid with the larger claw
bidentate, the smaller unidentate (viewed in one direction the second
or basal tooth of the larger claw seems to stand out laterally,
looking like three claws).
Wings: Mostly dark scales, with a few scattered white ones;
first fork-cell considerably longer and narrower that the second, its
base nearer the base of the wing, its stem as long as the cell, stem
593
of the second longer than the cell; third long vein very close to the
second; cross-veins thick, clouded, all three close together; fringe
with eight white spots; the scanty dark scales give the wings a
spotted appearance. Halteres yellow, knob black at the apex.
Length . 7 mm.
Habitat, Wadelai, Lado District, Sudan. (H. King, 8/6/11.)
Observations . Described from a perfect male taken from under¬
growth around trees on a grassy slope near the Nile. It was
pointed out as being a distinct species by Mr. King. It is a very
dark species, easily distinguished by the leg ornamentation.
Chrysoconops NOCTURNUS, nov. sp.
Head golden-yellow; proboscis and palpi golden-yellow, black
at their apices. Thorax shiny black, with scattered golden scales;
pleurae mostly brown, a long yellowish area before wing roots.
Abdomen entirely golden-yellow. Legs golden-yellow, femora with
some scattered dark scales and narrow dark apical bands; fore and
mid tibiae with scattered dark scales and a small dark apical band;
hind tibiae with narrow basal and broad median and apical dark
bands; fore and mid tarsi golden-yellow, unbanded, last two tarsals
dark; hind legs with metatarsi, first, second and third tarsals with
broad apical black bands, last all dark. Wings yellowish, all the
veins with dusky brown and dull yellowish scales. Male palpi
with three dark rings, legs darker than female and traces of dark
apical abdominal bands.
9 . Head : Integument greyish, clothed with golden-yellowish
narrow-curved scales and dark, thin, upright, forked scales; eyes
silvery. Antennae brown, basal segment pale. Palpi and
proboscis golden, black scales at their apices, with black chaetae
and some scattered black scales on the apical half of the proboscis,
also some black scales below, at the base (one specimen shows some
scattered dusky scales over the palpi).
Thorax : Shiny black, with scattered golden-yellow, thin,
narrow-curved scales, and black chaetae; prothoracic lobes
projecting, golden-yellow, roundish and mammillate. Scutellnm
black, shiny, with golden narrow-curved scales and dark border
bristles. Metanotum black. Pleurae deep brown with some
594
patches of flat white scales and a yellow, longish area at the
base of the wings; a short blunt process appears on each side of
the front region of the mesonotum. (Fig. 2. B.)
Fig. 2. Cbrysoconops nocturnus. n. sp.
A. * Prothoracic lobe. B. = Metonoul process.
Abdomen: Steely, entirely clothed with bright golden-yellow
scales and yellow hairs; venter golden yellow.
Legs: Golden-yellow, femora with some scattered black scales
and black apices, fore and mid tibiae with scattered black scales
and black apices, hind with narrow dark basal rings, a broad black
median and apical rings; fore and mid tarsals unbanded, the last
two dusky; hind tarsals with broad apical black bands; chactae
dark; ungues dark, equal and simple.
Wings : Yellowish with scattered dusky and yellow scales,
appearing mainly dusky in certain lights; first fork-cell longer and
narrower than the second, its base nearer the base of the wing, its
stem about one-third the length of the ceil; stem of the second
fork-cell not quite so long as the cell; posterior cross-vein about
twice its own length distant from the mid.
Halteres pale yellow.
Length . 5 mm.
S • Palpi with the last two segments nearly equal, the apical
one mostly black, apex of the next two dark; scanty hair tufts,
brown. Antennae banded yellow and brown with rich brown plume
hairs.
Legs : As in 9 , but more dark scales above on the femora and
tibiae.
Abdomen : With some dark metallic scales on the apices of the
segments, almost forming bands, apical segment dark, with dark
595
chaetae and dark claspers. Fore and mid ungues very unequal,
the larger uniserrate; hind ungues rather long, curved and simple,
all black.
Length. 5 mm.
Habitat. Bier Terab (Shambe to Wau road), Bahr-el-Ghazal
Province; Lau (Shambe to Wau road); Lau to Ateiba (Shambe to
Wau road) and Mayo to Melangot (Naam River), Bahr-el-Ghazal
Province. (H. King, 30/12/10, 3/1/11, io/i/ii.)
Observations. Described from three 9 *s and one c?. The
specimens were taken by Mr. King in a rest house after dark and
out of doors after dark. It is a vicious blood-sucker. This species
comes near C. nigrithorax , Theob., which it superficially resembles,
but it is distinct, as pointed out by Mr. King. The tibiae are not
all black, as in C. nigrithorax ; the proboscis has a few black scales
at the base, but is not deep black on the apical half, as in C. nigri
thorax. The marked prothoracic lobes also differ, and the blunt
lateral processes on the mesonotum are also characteristic.
Reedomyia SUDANENSIS, nov. sp.
Head dark, a black patch and then a pale patch on each side,
middle with golden scales and black upright scales; eyes silvery
and black; palpi, proboscis and antennae deep brown. Thorax
deep rich brown with scattered small golden scales; scutellum
silvery white; pleurae blackish-brown to brown with white puncta.
Abdomen deep blackish brown, unbanded, with small basal lateral
white spots; venter with basal yellow scales, the segments with
blackish apical borders. Legs deep brown, unbanded, hind femora
and tibiae with apical white spots, seen also in fore and mid legs
to some extent; chaetae golden.
9 . Head: Black and shiny with scattered narrow-curved pale
golden scales in the middle, with black upright forked scales, a
patch of flat black scales at the sides and then flat creamy white
ones. Eyes silvery around the upper border and down to the
antennae. Palpi black. Proboscis deep brown. Antennae deep
brown with pale pubescence and dark hairs, basal segment and base
of second paler.
Thorax : Deep rich brown, with small narrow-curved scattered
596
golden scales, a small patch of broader creamy ones in front of the
root of each wing; chaetae dark. Scutellum with flat silvery white
scales and numerous rather irregular dark border bristles.
Metanotum dark with a greyish sheen. Pleurae black, with patches
of white scales.
Abdomen : Black, unbanded, with small basal white lateral
spots, dark border bristles with dull golden apical reflections;
venter with basal creamy yellow areas, dark apical ones.
Legs : Deep brownish-black, unbanded, apices of femora and
tibiae white, especially in the hind legs, chaetae golden, showing
markedly against the dark legs; ungues of fore and mid legs equal
and uniserrate, of hind equal and simple.
Wings: First fork-cell longer and narrower than the second, its
base nearer the base of the wing than that of the second, its stem
about half the length of the cell; stem of the second fork-cell not
quite as long as the cell; posterior cross-vein about its own length
distant from the mid.
Halteres with pale stems and fuscous knobs, with some pale
scales on the latter.
Length. 4 to 4*5 mm.
Habitat. Yidu, Lado District, two 9’s; Khor Nambiri, Lado
District, one 9 ; Nyumbe, Lado District, one 9 ; Such River,
one 9 ; 12 to 14 miles from Hierallah on road to Bundle, Lado
District. (H. King, 22/3/11, 2/4/11, 27 and 31/5/11.)
Observations. Described from six 9's. The marked head
adornment is very noticeable. In one 9 there are traces of the
thoracic scales forming two dorsal spots, but the deep rich brown
with fine golden scale dusting is most noticeable in all six specimens.
It comes near R. bipunctata , Theob., but the scutellum is snowy
white, not pale yellowish, and the thorax has golden, not bronzy
scales; from R. biannulata , Theob., it can be told by the apex of
the abodmen not being white and from R. neobiannulata , Theob.,
bv having no basal white abdominal bands.
Mr. King's notes are as follows: —
‘ Khor Nambiri (Kapei to Yei), Lado District, 2/4/11. A small
khor containing pools, and shaded by trees and bushes.'
‘Nyumbe, Lado District, 27/5/11. Amongst undergrowth in
a grove by a khor.'
597
‘Yidu, Lado District, 31/5/11. From undergrowth on the bank
of a typical Glossina palpalis khor.*
( 12-14 miles from Hierallah on the road to Bundle, Lado
District, 22/3/11. In a deep, heavy timbered ravine/
Kingia MACULOABDOMINALIS, nov. sp.
Head black, silvery white on middle and sides; proboscis black;
palpi black, snow white tips; antennae deep brown, black at base
with white scales; eyes silvery. Thorax rich deep brown, a large
patch of snowy white on each side, a small one over the wings, and
lines of yellowish scales posteriorly; scutellum snowy white in some
lights, grey in others; pleurae deep brown with white puncta.
Abdomen deep black, unbanded, 5th to yth segments with
silvery white median spots, and with basal lateral white spots.
Legs dark brown, femora with traces of two white spots; first and
second tarsals only with narrow basal pale bands (fore and mid
legs).
$. Head: Black, clothed with flat black scales, flat white
ones in the middle and a patch of white ones on each side; chaetae
black. Palpi black scaled with snow white scales apically.
Proboscis black. Antennae deep brown, the basal segment with
flat white scales, the second segment with outstanding dark scales,
hairs deep brown.
Thorax : Shiny black with deep bronzy black, narrow-curved
scales, two large patches of snow white flat scales about the middle
of the mesonotum, a small patch of narrow-curved white ones just
in front of the roots of the wings, a short line of pale creamy scales
running up to the bare space before the scutellum, and a line of
yellowish scales on each side parallel to it and reaching the
scutellum; chaetae black. Scutellum black, the flat scales are
glassy and snow white in some lights, grey to almost black in
others; border bristles blackish brown, four to the rtiid lobe.
Metanotum deep chestnut-brown. Pleurae deep brown with silvery
white puncta.
Abdomen : Black with basal snow white spots, the fifth with two
median silvery white spots, the sixth with one median snow white
spot, the seventh with the spot almost basal, a small one also on
the apex; border bristles deep brown; venter dark brown.
598
Legs : Blackish-brown, femora with two white spots; first and
second tarsals of fore and mid legs with basal creamy white bands,
others dark in fore and mid legs (hind legs missing); ungues equal
and uniserrate.
Wings: First fork-cell longer and narrower than the second,
its stem more than half the length of the cell, stem of the second
fork-cell nearly as long as the cell; posterior cross-vein rather more
than its own length distant from the mid; scales brown, rather
large.
Length. 4 mm.
Habitat . Khor Kokbwa (Yei to Abba), Lado district.
(H. King, 13/4/11.)
Observations. Described from a single 9 taken by a rocky,
shady stream; the hind legs missing. A very marked species
readily distinguished by the thoracic and abdominal adornment.
Aedimorphus QUINQUEPUNCTATA, now sp.
Head black and silvery white; palpi, proboscis and antennae
deep brown. Thorax rich brown with four silvery white spots and
a small fifth white patch before the bare space in front of the
scutellum, which is also silvery white scaled; pleurae deep brown
with silvery white puncta. Abdomen dusky black, unbanded, with
basal lateral silvery white spots; venter with basal silvery white
lateral spots also. Legs dark, unbanded, with venter of femora
pale at the base, especially on the hind legs where most of the
venter is pale, femora of hind legs, each with a snow white apex
and a spot before the apex.
9 . Head : Black with a small median area of dull creamy
narrow-curved scales, with black upright forked scales, a patch of
flat snow white scales on each side in front, then flat black scales,
then white and then black ones again. Clypeus , proboscis , palpi
and antennae deep brown to almost black.
Thorax : Dull black with small, narrow-curved, dull brown
scales and four patches of small flat snow white scales, two in
front near the head, two in the middle of the mesonotum, also a
fifth patch before the bare space in front of the scutellum, and a
small spot below in front of each wing; chaetae black. Scutellum
599
black with snow white flat scales forming three patches, border-
bristles black, four large ones to the mid lobe. Metanotum deep
black; pleurae rich brown with six snow white puncta.
Abdomen : Black with dull golden border bristles and basal
’snow white lateral patches, the scales somewhat outstanding; venter
black with basal snow white lateral spots.
Legs : Almost black, femora and tibiae with apical white spots
most prominent on the hind legs, each of the hind femora with a
round snow white spot near the apex and whitish on most of the
venter; ungues of fore and mid legs equal and uniserrate, hind
equal and simple.
Wings : With a white scaled spot at the base and with the
first fork-cell slightly longer, but no narrower than the second fork¬
cell, its stem more than half the length of the cell; stem of the
second fork-cell about two-thirds the length of the cell; posterior
cross-vein less than its own length distant from the mid; sixth
long vein markedly curved. Halteres with ochreous stem and
ochreous and dusky knobs.
Length . 3*5 mm.
Habitat . Alenga and Matalee, Lado District. (11/5/11 and
28/5/11, H. King.)
Observations . Described from two $'s. It comes near
Acdimorphus punctithorax , Theob., but differs in having five, not
six, thoracic white spots.
CULICELSA CENTROPUNCTATA, nov. sp.
Head brown with pale scales and two dark areas on each side
separated by a line of white; palpi, proboscis and antennae deep
brown. Thorax with brown and silvery white scales, the latter
forming two spots in front, a median and more or less pronounced
posterior area; scutellum entirely white scaled. Abdomen black
with median white basal patches and basal white lateral spots;
venter mostly creamy scaled. Legs black, with pale chaetae,
especially marked on the tibiae, femora and tibiae with apical white
to yellow bands, tarsi with narrow basal white bands to all the hind
segments, and all but the last two in the fore and mid.
9. Head : Clothed with almost white narrow-curved scales in
the middle and dark, thick, upright forked scales, then a few flat
white scales, then a black patch, then white, then black again;
chactae long and deep brown, except for some golden ones
l)ctwcen the eyes. Palpi , proboscis and clypcus almost black.
Antennae black with pale internodes, basal segment dark and
testaceous, the second with flat black scales; verticillate hairs
black.
Thorax : Deep brown with mixed brown and silvery white
narrow-curved scales, the latter forming two spots in front, a border
around the mesothorax near the head, denser behind, and as a line
above each wing; chaetae long and black. Scntellum with narrow
white curved scales. Metathorax black. Pleurae deep brown
with patches of flat white scales.
Abdomen: Black, the segments with median basal patches of
white scales, and patches of basal lateral white scales which
become median apically, posterior border bristles golden; venter
mainly creamy scaled.
Legs: Almost black, femora and tibiae with apical pale spots,
base of first and second fore and mid tarsals with white bands; in
the hind legs all the tarsals have basal white bands, bristles dark
and golden; fore and mid ungues equal and uniserrate, hind equal
and simple.
Wings : With short fork-cells, the first longer and narrower
than the second, its base a little nearer the base of the wing, its
stem nearly as long as the cell; stem of the second longer than the
cell; posterior cross-vein twice its own length distant from the mid.
Halteres all pale ochreous.
Length. 4 mm.
< 3 . Head : Pale scaled w'ith tw ? o dark patches of flat scales
surrounded by white flat scales. Palpi a little longer than the
proboscis, black, with the last two segments nearly equal and with
small basal w'hite bands, a creamy band at the base of the third
segment and a pale area at their base, on the last two segments arc
brown hair-tufts on one side, and on the apex of the penultimate
is a long golden chaeta. Antennae banded brown and grey, plume
hairs flaxen brown.
Thorax , abdomen and legs as in the 9 ; fore and mid ungues,
unequal, uniserrate, hind equal and simple.
Length . 4*5 mm.
Habitat. Alcholi, Lado District and River Mewri, Mongolia
Province. (25/5/11, 9 ; 21/3/11, c?, H. King.)
Observations . Described from a perfect ( 5 * and 9 » although
taken in different places undoubtedly the same species. It is a
most marked species and is related to those so far placed in the
somewhat obscure genus Culicelsa y Felt, obscure only for the fact
that we cannot fix any definite characters, yet all the species
included have a very marked similar appearance.
Heptaphlebomyia KINGII, nov. sp.
Head black with pallid scanty scales, proboscis ochreous black
at the base and apex; antennae and palpi dark brown. Thorax
rich brown, somewhat paler in the middle, two snow white spots
in front near the head and two on the middle of the mesothorax;
scutellum snow white scaled; pleurae deep brown with snow white
puncta.
Abdomen deep brown, unbanded, with basal lateral white spots;
venter with pale basal bands. Legs unbanded, deep brown, femora
and tibiae with apical white spots. Wings with seventh vein very
close to the wing border.
9. Head : Black with pale creamy, narrow-curved scales in
the middle, becoming white in front, creamy flat lateral scales,
white at eye border near the white narrow ones; upright forked
scales black. Antennae , palpi and clypeus almost black.
Proboscis ochreous brown, black at the base and apex.
Thorax \ Black with narrow-curved bronzy brown scales, two
median bare parallel lines, four round spots of snow white narrow-
curved scales, two in front near the head, two somewhat larger and
wider apart about the middle of the thorax; the scales become
paler towards the scutellum, being pale golden in some lights;
chaetae brown. Scutellum black with dense snowy white curved
scales, deep brown border bristles, six to the mid lobe; metanotum
black with grey reflections. Pleurae deep brown with patches of
rather long, flat, white scales.
Abdomen : Steely, clothed with black scales, with small snow
white basal lateral spots; border bristles brown with golden
reflections apically; venter black with basal white bands.
602
Legs: Dark brown, unhanded, femora pale at the base, apices
of femora and tibiae white on the hind legs, traces in the mid,
scarcely perceptible in the front pair; ungues equal and simple
Wings : With brown scales; first fork-cell considerably longer
and a little narrower than the second fork-cell, its base nearer the
base of the wing, its stem Idss than one-fourth the length of the
cell, stem of the second posterior not quite as long as the cell;
posterior cross-vein not quite twice its own length distant from the
mid; the seventh scaled vein close to the inner border of the wing.
Halteres pale with fuscous knobs.
Length. 4 mm.
Habitat. Nyumbe, Lado District; Alenga, Lado District.
(27/5/11, Nyumbe; 28/5/11, Alenga.)
Observations. Described from two perfect 9*s. It can he
distinguished from the allied H. argentcofunctata , Ventrillon, by
the abdomen having only small basal lateral spots and not
ornamented as in VentrilIon’s species from Madagascar; the
antennae are also black in the 9> not yellowish, and the thoracic
adornment differs. The scaled seventh vein is well marked in
both 9 *s. One was taken from a small hole, containing water, in
a tree, the other from tall grass near a khor.
603
THE PROBABLE IDENTITY OF
TRYPANOSOMA CONGOLENSE
(BRODEN) AND T. NANUM (LAVERAN)
BY
B. BLACKLOCK
AND
WARRINGTON YORKE
(From the Runcorn Research Laboratories of the Liverpool
School of Tropical Medicine )
(Received for publication 28 November , 1913)
In our paper on the identification of the more important
mammalian trypanosomes, we have regarded T. dimorphon (sensu
Laveran and Mesnil), T. confusum (Montgomery and Kinghorn) and
T . pecorum (Bruce) as synonymous with T. congolense , which was
first described by Broden in 1904. In the same year Laveran
described a similar parasite found by Balfour in the Sudan, under
the name of T. nanum. These parasites are identical morpho¬
logically in that they are both short aflagellar trypanosomes
measuring 8 to 19/4 in length. The sole distinguishing feature is
their effect on small laboratory animals, T. congolense being
described as pathogenic for monkeys, dogs, rabbits, guinea-pigs,
rats and mice, while T. nanum is considered to be incapable of
infecting these animals. The object of this paper is to examine
the evidence upon which this distinction is based and to decide
whether it is sufficient to warrant such a differentiation.
In previous papers* a description has been given of two
trypanosomes which were present in the blood of a naturally infected
horse sent over to this country from the Gambia. One of the
parasites was unquestionably T. vivax : concerning the identity of
* Yorke and Blacklock. The trypanosomes found in two horses naturally infected in the
Gambia. Annals of Tropical Medicine & Parasitology, 1911, Vol. V, p. 413.
Blacklock. The trypanosomes found in a horse naturally infected in the Gambia.
A double infection. Annals of Tropical Medicine & Parasitology, 1912, Vol. VI, p. 107.
the other there was, however, considerable doubt. Morphologically,
this parasite was indistinguishable from T. dimorphon (Laveran
and Mesnil) and T. nanum. As we finally succeeded in infecting
small animals with the trypanosome, it was eventually decided that
it was T. dimorphon ( T . congolense).
Both the parasites, which were separated fortuitously, one from
the other as already described, have been kept in experimental
animals for a period of 18 months. The results of artificial passage
of the short aflagellar parasite from animal to animal are so
interesting and suggestive that we have decided to describe them
in some detail.
The sub-inoculations made with this parasite from the time of
its isolation until the 51st generation are given in genealogical form
in the table. A study of this table reveals two facts, viz.: —
(1) Most of the early inoculations failed to infect, whereas the
later were invariably successful.
(2) The course of the infection in the earlier successful cases
was chronic, whereas that in the later instances was acute. Thus,
if we consider the animals used in the second to the fourth
generation, we find that these comprise 8 rats, of which 3 were
positive and 5 negative; 4 mice, of which 2 were positive and
2 negative; 3 rabbits, 2 positive and 1 negative; 4 guinea-pigs,
2 positive and 2 negative; and 3 goats, 1 positive and 2 negative.
Again, if the duration of the disease in the earlier rats be
compared with that in the later experiments, the contrast is very
striking. For example, the average length of life of the first ten
rats from the fifth generation to the fourteenth was 88*6 days,
whereas that of the last ten rats, comprising the 42nd to the 51st
generation, was only 8*6 days.
It is clear, therefore, that by passage of this parasite through
laboratory animals the trypanosome has been changed from one of
uncertain and chronic pathogenicity to one of great virulence.
This fact seems to us to be one well worthy of remark. That
artificial passage of a strain through a series of animals does some¬
times alter its virulence for that species is well known. The results*
• Warrington Yorkc. On the pathogenicity of a trypanosome from a case of Sleeping
Sickness contracted in Rhodesia. Annals of Tropical Medicine <SL Parasitology, 1910,
605
obtained by different workers with T. gambiense are illustrative of
this point.
These observations appear to us to have some significance for
the nomenclature of the parasite. T. congolense and T. nanum are
identical morphologically, they are both spread by the same species
of tsetse fly and infect the insect in precisely the same manner.
The usual way of deciding with which parasite cattle or
antelope known to harbour a short trypanosome in their blood are
infected, is by sub-inoculation of rats or some other convenient
laboratory animal. As a rule, these animals are not too plentiful
in the tropics and one or two must suffice for the diagnosis. That
inoculation of one or two small animals may not afford any
conclusive evidence as to whether or not the trypanosome is
pathogenic, is at once realised from observing the results of the
earlier inoculation of the parasite from our horse. Had the number
of our experimental animals been limited, we should probably have
designated the parasite T. nanum . Further experiments, however,
showed that the trypanosomes could be made acutely pathogenic
to rats.
This opens up the question as to whether there is really any
difference between T. congolense and T. nanum . It is interesting
in this connection to refer to the observations of other workers.
Writing in 1911 on a short aflagellar trypanosome obtained from
ponies naturally infected in Togoland, Weissenborn* states that
the parasite was of inconstant virulence. It was most virulent for
mice, but slightly so for rabbits, rats and monkeys. Only a small
proportion of rats were susceptible, whilst guinea-pigs were
absolutely refractory. Morphologically the parasite T. frobeniusi
closely resembled T. congolense.
The Sleeping Sickness Commission of the Royal Society! write:
‘If T . pecoruMy which is usually more or less infective in the
monkey, dog and rat, lives for some time in the blood of the goat,
it loses its power of infecting other animals. This has given rise
to the erroneous idea that a separate species— T. nanum exists.*
• Weissenborn, E. Beitrag zur Kenntnis dcr kurzgcisscligcn Trypanosomcn. Archiv.
f. Schiffs- und Tropen-Hygiene 1911, p. 477.
t Bruce, Harvey, Hamerton and Lady Bruce. The susceptibility of various animals to
T. simiae. Roy. Soc. Proc., 1913, Vol. 87, p. 49.
6o6
The Belgian Sleeping Sickness Commission* found that aflagellar
trypanosomes from naturally infected dogs will not always infect
guinea-pigs and rats.
It is thus evident that workers in the field have found that short
aflagellar trypanosomes, morphologically identical with T. con-
golense and T . nanutn , are of uncertain pathogenicity for the
smaller laboratory animals.
As the result of our investigations and of those of the authors
mentioned above, we can see no evidence which would justify
distinguishing one from the other on the ground of pathogenicity.
In the present state of our knowledge we can only conclude that
T. congolense and T . tiamim are the same parasite.
* Rodham. Pons, Vanden Branden. and Hequaert. Rapport «ur lc» Travaux de la Minion
Scientific du Katanga. Octobre 1910 a Septcmbre 1912.
HERPETOMONAS STRATIOMYIAE,
n.sp.j A FLAGELLATE PARASITE OF THE
FLIES, ST RATIO MV I A CHAMELEON
AND S. POT AMID A, WITH REMARKS
ON THE BIOLOGY OF THE HOSTS
BY
H. B. FANTHAM, D.Sc. (Lond.), B.A. (Cantab.)
(LECTURER ON PARASITOLOGY, LIVERPOOL SCHOOL OF TROPICAL MEDICINE)
AND
ANNIE PORTER, D.Sc. (Lond.), F.L.S.
(QUICK LABORATORY, CAMBRIDGE)
(Received for publication 19 A November, 1913)
Plate XLI
CONTENTS
PAGE
I. Introduction ... ... ... ... ... ... ... 609
II. The Biology of the Host Flies, Strattotnyia chameleon and S. pot amid a ... 610
III. Material and Methods ... ... ... ... ... ... 612
IV. The Life-History or Herpetomonas stratiomyiae
(A) The Herpetomonas in the Larva of the Fly ... ... 612
(B) The Herpetomonad during Pupation ... ... ... 615
(C) The Herpetomonad in the Imago ... ... ... 616
V. Mode or Inpection ... ... ... ... ... ... 617
VI. Concluding Remarks ... ... ... ... ... ... 617
VII. Summary ... ... ... ... ... ... ... ... 618
References ... ... ... ... ... ... ... 619
Explanation of Plate ... ... ... ... ... ... 620
I. INTRODUCTION
The interesting flies, Stratiomyia* chameleon and S. potamida ,
are fairly common in certain districts where the drainage of the land
is poor and where boggy areas with somewhat rank vegetation occur.
Both the larva, pupa and imago of the insects are sometimes
parasitised by a small Protozoon, belonging to the genus
* The original generic name of the host flies was Stratiomys. Many modern authorities
prefer the f >rm Stratiomyia , which has been adopted in this paper. The Stratiomyia were
obtained near Cambridge, and one local name for them is Chameleon flies.
6 io
Herpetomonas, and presenting certain differences from other
common Herpetomonads that warrant its inclusion as a new species.
The chief interest of the parasite lies in its relation to the life-
history of the host, and thus a brief note on the biology of the
hosts is considered advisable.
We do not propose to enter into great cytological detail
regarding the structure of the flagellate, nor to engage in fruitless
discussion as to the nomenclature of such parasites, but rather to
consider the flagellate in relation to the life-history of the host.
The relation of insect flagellates to their hosts* life-cycle is a most
interesting though difficult subject, on which little or nothing has
been done in most cases up to the present.
II. THE BIOLOGY OF THE HOST FLIES, STRATIOMYIA CHAME¬
LEON AND 5. POTAM/DA
(a) Habits. The larvae of these insects are very characteristic,
worm-like, legless organisms, armed with powerful jaw's by which
they drag the body forwards, progression being aided by bristles
attached to each segment. The larger larvae examined varied
from 2 inch to 2 \ inches in length at will, as the abdomen can be
retracted telescopically. Their colour varies w'ith that of the mud
and decaying vegetation among which they live, yellowish, brown
and green being common hues. Twenty-five to thirty respiratory
tail filaments are present and arc applied by the larva to the
surface film, so that in deep water the larva is suspended vertically,
head downwards.
The pupae are found near to the surface in the mud fringing
the pond or stream. The larval skin is retained and the much
smaller pupa is enclosed within it. The period of pupation is
short.
The imago is somewhat bee-like and is blackish in colour with
bright yellow markings. Eggs are laid on aquatic plants just above
the level of the water, and the larvae either escape into the water,
or are hatched beneath the surface into which the plants finally
fall.
( b ) Food. The food of the imago seems to be sweet vegetable
material. The larvae, so far as can be ascertained by' direct obser¬
vations extending over three years, are vegetable feeders. The
contents of the alimentary tracts of those examined have never
contained blood or other recognisable animal matter such as muscle
or chitinous tissue that could be obtained from insects or Crustacea.
Small organisms such as Protozoa may be ingested, but no proof
that such was the case occurred. A typical set of examinations of
50 larvae from 10 different localities, representing about one-third
of the total number examined, gave the following results: —
From the first locality, seven larvae varying from { inch to 2\ in. long, were obtained.
Two were infected with Hcrpeiomonas, one infected larva was in. long, the other in.
The large vegetation of the locality comprised grass. Veronica beccabunga , Myosotis
palustris, various algae, and many rotting leaves. The contents of the food canals of
the larvae were portions of Chara , Cladofhora , Spirogyra and Zygnema.
Ten larvae from the second locality were from i in. to 2 in. long. None were
infected. Grass and Veronica beccabunga constituted the larger vegetation. There were
fewer' large algae than in the first locality. The alimentary canals of these larvae
contained algae, chiefly species of Clostcrium, Penium , Micraslerias and Cosmarium.
Six larvae from a third place varied from in. to 2.1 in. long. Like the previous
set, they were uninfected. Their gut contents consisted almost entirely of Goninm sp.,
and the larger vegetation on the spot comprised grass, Myosotis and Common Hemlock.
A small pond, whose bordering vegetation consisted of grass and Myosotis , and
in which some fine Hottonia grew, yielded seven larvae. These contained many algae,
chiefly species of Scenedesmus and Selenastrum . No paiasites were found in the larvae.
Six larvae were obtained from the fifth locality. All were vigorous, and varied
from 1 in. to 2\ in. in length. Grass, Myosotis and algae were abundant. The food
passages of the larvae contained algae, chiefly species of Navicula, Diatoma , Synedra ,
Gyrosigma and Nitzschia. No herpetomonads were found.
Some of the smallest larvae were obtained from the sixth localitv. They varied
from in. to 2 in. in length. Seven were examined. A larva, 1 in. long, had a slight
infection with Ilcrpctomonas stratiomyiae , the rest were normal. Rotting leaves
occurred in the water and hemlock, Veronica bcceabunga and grass constituted the
larger vegetation. Algae, chiefly species of Navicula, Denticula and Nitzschia , together
with fragments of leaves, were found in the alimentary canals of the larvae.
Three larvae only were obtained from the seventh locality, w'hose vegetation was
the same as that of the sixth. The gut contents of the larvae also were similar. None
were infected.
One specimen, 1$ in. long, obtained from a small brook partly choked with
Potamogeton crispus and large algae, contained no parasites. Its gut contents included
species of Scenedcsmus and Cosmarium . together with fragments of Chara .
One larva, 2 in. long, was obtained from a ditch fringed with willowherb. Its gut
contained numerous specimens of Cosmarium and a very few Herpetomonads also were
present.
Two larvae from the tenth locality were uninfected. The chief plants growing in
the mud were willowherb and water dropwort, while the food contents of the larvae
consisted of species of Cosmarium and Navicula.
The presence of infected larvae coincided to some extent with
the quantity of decaying matter that was present; but sometimes
larvae obtained from the foulest sources (not those detailed) were
as free from infection as those from relatively uncontaminated
places. Larvae were in greatest abundance where much decaying
material was present.
6l 2
Altogether, nearly 150 larvae were examined, and the per¬
centage infected with Herpetomonas was only about 3. The
percentage of infection in the pupae examined was somewhat higher.
III. MATERIAL AND METHODS
The larvae, pupae and imagines of Stratiomyia , comprising both
bred insects and those caught at large, have been used during this
investigation. The Herpetomonas has been found most abundantly-
in the digestive tract of the larva. The alimentary canal has been
dissected wherever possible, placed in physiological salt solution
and examined in serial teased portions. The pupal condition
usually prevented dissection of the alimentary canal, but smears
of the central part of the body were sufficient to allow of identifica¬
tion of the parasite. Much time has been spent in observing the
living Herpetomonas. Osmic acid followed by absolute alcohol,
Bouin’s fluid and Flemming’s solution have been used for fixatives,
and Giemsa’s solution, iron haematoxylin and glycerin haematin
were used as stains. The paraboloid condenser and stereoscopic
oculars have also been used and found of much service in
determining depths of granules, chromatin particles and similar
structures.
V. THE LIFE HISTORY OF HERPETOMONAS STRATIOMYIAE
The life-cycle of the herpetomonad may be divided into two
main phases: (1) a resting phase, (2) an active, multiplicative
phase. The resting phase again can be sub-divided according to
the period at which it is present in the host. Infection is by the
contaminative method, and freshly ingested resting forms of the
parasite about to develop into the active form are best described as
the preflagellate stages, while the forms produced by the preparation
of the full flagellate for extra-corporeal life are more accurately
described as post-flagellate forms. Naturally the post-flagellate
form produced in the first host becomes the pre-flagellate organism
in the second host.
A. The Herpetomonad in the Larva of Stratiomyia
The preflagellate stage of Herpetomonas stratiomyiae differs
from that of most herpetomonads in being elongate oval in shape
(PI. XLI, figs. 1, 2). Its length varies from 5-5^ to 8 ft, while
its breadth is about 3 p. The cytoplasm is clear in life or slightly
granular. The nucleus shows as an oval, refractile vacuole, the
blepharoplast as a bar-like rod. They both lie relatively near the
surface. Sometimes the end of the body near the blepharoplast
appears somewhat more refractile in life than the distal end. Such
an area, when stained, proved to be the chromatophile vacuole¬
like area from which the flagellum originates. The nucleus presents
a definite nuclear membrane, seen best in haematin-stained pre¬
parations. The chromatin varies in its arrangement, sometimes
appearing as a more or less central concentrated karyosome (fig. 2),
at other times being scattered as granules in the nucleoplasm (fig. 1).
The bar-like blepharoplast stains deeply. The formation of a
flagellum occurs very rapidly, and hence it is difficult to find stages
in which the chromatophile area is present unless the host is
dissected just at the time preceding the flagellation of the parasites.
Division occurs among the preflagellates (figs. 4, 5), and will be
described later.
The flagellates (figs. 10-17) vary in size, the length including
the flagellum being from 26*6/1 to 57/1, while the breadth is from
2/1 to 3*6 p. The flagellum itself may occasionally reach 38/4 long.
The non-flagellate or posterior end of the flagellate is elongate but
somewhat blunted; the flagellar or anterior end is somewhat
rounded, with the flagellum projecting from it. As with the
preflagellate forms, the nucleus is oval and shows cyclical develop¬
ment. Relatively young flagellates, or flagellates that have formed
rapidly from the preflagellate forms, often possess a vesicular
nucleus with a central (figs. 15, 16) or slightly excentric (fig. 12)
karyosome. Older parasites, those of slow growth and forms about
to divide, have their nuclear grains of chromatin evenly distributed
(figs. 11, 14). The blepharoplast is curved, rod-like or oval, and,
except in dividing forms, is homogeneous in structure. Chromatoid
granules may be present in some flagellates, usually forming small
grains in the post-nuclear part of the body (figs. 11, 13).
Multiplication . The increase in numbers of the parasites within
the host is brought about by longitudinal division, which can take
place in either the preflagellate (figs. 3-5) or flagellate (figs. 18-22)
stages. The division of the preflagellates is initiated by that of
the blepharoplast, which becomes dumb-bell shaped (^fig. 3),
showing two concentrations of chromatin, one at each end. The
heads of the dumb-bell remain attached to one another for some
time by a strand of chromatin. The constriction of the nucleus
rapidly follows that of the blepharoplast. The future flagellar end
then becomes cleft (fig. 4), and the cleft extends backwards. As
soon as parts of the two daughter organisms are free, rapid move¬
ments occur, each free part diverging from the other and twisting
on itself as it does so, until the two organisms come to lie in almost
a straight line (fig. 5). Violent movements precede the final
separation, and this is usually succeeded by relative quietness on
the part of both the daughter forms, which usually do not move
away until some seconds and, occasionally, minutes have elapsed.
When movement occurs it is not active as with the flagellates, but
consists of slow undulations of the body, which gradually propel
the daughter forms forwards.
The stages of division of the flagellate (figs. 18-22) are like
those of the preflagellate form, but the division of the blepharoplast
and flagellum takes place almost simultaneously (figs. 19, 20), while
nuclear division is often delayed, and at times, cleavage of the
cytoplasm has commenced (fig. 20) before the nucleus has com¬
pletely divided. The movements of the two parts of the dividing
organism (figs. 21, 22) are very active, and the final separation is
effected more quickly than that of the dividing preflagellate, the
lashing of the daughter flagella being of material aid. The two
newly-formed individuals swim away rapidly after separation.
Examination of stained specimens has been confirmatory in all
respects of the above observations made on the living organisms.
The formation of a post-flagellate form (figs. 23-36) is brought
about by the gradual retraction and absorption of the flagellum
(figs. 23-27) and the concentration of the body until it forms an oval
or rounded form, which secretes a thin, closely adherent cyst around
itself. When the assumption of the post-flagellate form is com¬
plete, there is no trace of the flagellum remaining as such. Even its
root (or rhizoplast) disappears, the chromatin of it appearing to be
dissolved and concentrating, in some cases, in a small part to form
a chromatoid area, as well as diffusing generally through the body
substance, as shown by the staining reactions of the cyst. The
post-flagellate is Leishmania-like (figs. 32-36), of a somewhat
6i 5
elongate type. The nucleus may, or may not, show a karyosome,
for as with the flagellate, the structure of the nucleus varies. If
there is a karyosome in being at the time of encystment, the nucleus
of the post-flagellate shows the same feature (figs. 34, 36). Should
encystment follow rapidly on division, the nucleus is almost
homogeneous (figs. 33-35). Much discussion has arisen recently as
to the existence of specially differentiated portions of the blepharo-
plast, described variously as karyosomes and centrioles. In some
cases, by the use of iron-haematoxylin, some granules have been
demonstrated, but it is admitted that the existence of these granules
depends for its demonstration on the degree of removal of the
stain by the iron alum. They are not found when other stains are
employed. The basing of argument on the fleeting appearances
produced by too great or too little differentiation—the degree of
differentiation that is ‘accurate* being an absolutely individual
opinion—is merely vexatious, and does not tend to the advancement
of knowledge on really scientific lines.
B. The Herpetomonad during Pupation
As the period for pupation of the host approaches, the post-
flagellate forms of the parasite leave the gut of the larval host and
pass out with the faeces. Rapid flagellation of any preflagellates
that are present occurs, together with division of the flagellates.
The organisms seem to collect in the middle-third of the gut and
place themselves with their flagella towards the centre of its lumen,
their aflagellar ends being in contact with its wall. The latter is
very frail at this time, and the flagellates pass through it with ease.
They thus reach the haemocoel, and there they swim for a short time
in their normal position with their flagella forwardly directed.
Division occurs but infrequently. The flagellates (figs. 23-25)
gradually slow their movements and then rapidly concentrate their
substance (fig. 27). They soon become typical, post-flagellate
forms (figs. 27-31), with distinct nuclei and blepharoplasts, and very
thin cyst walls. These organisms are, perhaps, somewhat more
frail-looking than those found in the rectum and faeces of the larva
(figs. 32-36). At first they are intermingled with the numerous
wander cells found in the pupae at this time, but they seem to prefer
the more fluid parts of the host. The result is that, in a late pupa
6i6
in which the musculature has become fairly well developed, the post-
flagellates lie in the more fluid medium, internal to the region of
differentiated muscles. At times, they have been found crowded
together roughly at the centre of the pupa, and seem to be united
temporarily by gelatinous material. When the differentiation of
the alimentary canal is in progress, the parasites in most cases
become surrounded by it and can be found attached by the
gelatinous secretion to the cells of the gut. They remain thus until
the emergence of the imago. Four out of fifteen pupae were found
infected, but in one only was the infection heavy.
C. The Herpetomonad in the Imago
The number of infected imagines is much smaller than that of
infected larvae, though neither are abundant. The occurrence of
larvae in numbers in one district enables them to become
infected with post-flagellates from their neighbours with a certain
amount of ease, as they swallow the cysts with their food. But all
infected larvae do not give rise to infected pupae, while the latter
may have but a slight infection that fades out and disappears in
the adult. The habits of the imagines do not readily allow of them
acquiring infection ab initio , and consequently the numbers found
infected have been extremely small. The parasites have been found
in all stages. Non-flagellates, like those in the pupa, have been
found in the gut near the junction of the thorax and abdomen.
They rapidly form flagellates that spread quickly throughout the
whole length of the gut, while post-flagellates occur in the rectum
and faeces. No stages of the parasite have been observed up to
the present in the genital organs and but few in the haemocoel of
the host. When there is haemocoelic infection, there are two
possible means of origin : (i) the parasites may have remained there
from the pupation period, or (2) they may have penetrated from the
gut as flagellates. As they are capable of moving and developing
fully in the haemocoelic fluid, hereditary infection is possible,
though we are not in a position to pronounce on this with certainty,
owing to the small number of infected flies that have been examined
and to the difficulty of obtaining eggs. We hope to continue this
part of the work in the future.
6i 7
V. MODE OF INFECTION
The mode of infection is contaminative originally. The larvae
acquire post-flagellate forms of the Herpetomonas with their food.
The cysts may have been derived either from already infected larvae
or from the dejecta of adult flies. The faeces of a larva on one
occasion contained active flagellates as well as post-flagellate forms.
As the flagellates lived for some hours in ordinary water, it is
possible that they could do so in nature, and act as an additional
source of infection of new hosts.
The parasite can persist through the metamorphosis of the host
and the imagines thus emerge infected. It is possible, though we
think it is probably exceptional, that an adult insect may become
infected by sucking plant juices contaminated with excrement from
other imagines, and thus acquiring the post-flagellate stages of the
parasite.
VI. CONCLUDING REMARKS
The flagellate parasite of Stratiomyia chameleon and S. potamida
is a member of the genus Herpetomonas as originally defined. It
differs from H. pediculi (Fantham, 1912) from the body louse.
Pediculus vestimenti , in its characteristic, oval, preflagellate form.
Herpetomonas jaculum from Nepa cinerea is similar to H. stratiom -
yiae in appearance, but again the preflagellates of H. stratiomyiae
are distinctive, the same feature differentiating the parasite from
H. lygaei and H . culicis . It also shows differences from H . muscae
domesticae y H . ctenocephali , H. ctenophthalmi and other known
Herpetomonads. In consequence of these morphological differences,
and as it is the first time that such a parasite has been recorded from
the Stratiomyidae, the organism has been named Herpetomonas
stratiomyiae. We believe that this account is the first detailed
record of the behaviour of an insect flagellate during the pupation
of its host, a feature which gives a special interest to Herpetomonas
stratiomyiae .
The study of the life-cycle of such herpetomonad flagellates of
insects is of the utmost importance in view of the recent experiments
of Laveran and Franchini (1913) on the successful experimental
inoculation of H. ctenocephali (Fantham, 1912), from the gut of
the dog-flea, into mice and other mammals. Such researches suggest
an experimental leishmaniasis in the making.
6i8
VII. SUMMARY
r. Herf'Ctomonas stratiomyiae , n. sp., is a parasite of the
larvae, pupae and imagines of the flies, Stratiomyia chameleon
and S. pot amid a.
2. The herpetomonads present three stages in their life-history,
(a) a preflagellate stage, (6) a flagellate stage and (c) a post-
flagellate stage.
3. Preflagellate forms are characteristically oval, with elongate
nucleus and distinct blepharoplast. The flagellates vary from
26 6 m to 57 ac in total length. The post-flagellates, as found in the
hind gut of the larvae, are oval, with somewhat thicker walls than
the preflagellates.
4. Multiplication is by longitudinal division and can occur in
any phase of the life-history of the organism.
5. Just prior to pupation, the flagellates migrate from the gut
of the larva into the haemocoel where they gradually become
motionless, withdraw their flagella and assume the post-flagellate
form. They collect in the more fluid parts of the body of the
pupa, and, ultimately, as a result of this, they become enclosed in
the gut of the adult.
6. The imago, if formed from an infected pupa, usually
emerges from the pupa case infected with non-flagellate forms of
the herpetomonad. The development of the non-flagellate into the
flagellate form is rapid, and soon all stages of the organism are
present.
7. Infection of the larvae is contaminative, that of the pupa is
transmitted from the larva, while the imago may retain the pupal
infection or may possibly acquire it ab initio by ingesting
contaminated food.
REFERENCES
Further references will be found in some of the memoirs cited.
Fantham, H. B. (1912). Herpetomonas pediculi , nov. spec., parasitic in the Alimentary Tract
of Pediculus vesttmentiy the Human Body Louse. Proc. Roy. Soc., B, Vol. LXXX 1 V,
pp. 505-517. One plate.
- (191 a). Some Insect Flagellates and the Problem of the Transmission of Leisbmania.
Brit. Med. Joum., November 2, pp. 1196-1197.
Laviran, A., and Franchini, G. (1913)- Infections experimentales dc Mammiferes par de9
Flagclles du tube digestif de Ctenocepbalus canis et d’ Anopheles maculipennis. Compt.
Rend. Acad. Sci., VoL CLVII, pp. 744-747.
Pi».TTOV, W. S. (1908). Herpetomonas lygaei. Arch. f. Protistenkunde, Bd. XIII, pp. 1-18.
One plate.
-(1912). Studies on the Flagellates of the Genera Herpetomonas, Crithidia and Rhyn-
choidomonas. No. 1, The Morphology and Life-History of Herpetomonas culicis ,
Novy, MacNeal and Torrey. Sci. Mem. Govt. India, No. 57, pp. 1-21. One
plate.
Porter, A. (1909). The Life Cycle of Herpetomonas jaculuniy Leger, parasitic in the Alimentary
Tract of Nepa cinerea. Parasitology, Vol. IV, pp. 237-254. One plate.
620
EXPLANATION OF PLATE XLI
All the figures were outlined with an Abb6-Zeiss camera lucida,
using one-twelfth inch oil immersion lens and compensating ocular
No. 8. Magnification 1,500 diameters, approximately.
Figs. 1-22. Illustrate the preflagellate and flagellate stages found
in the larva.
Figs. 1-9. Illustrate the development of Herpetomonas straliomyiae
from the preflagellate to the flagellate stage.
Fig. 1. Preflagellate form with nuclear chromatin evenly
distributed.
Fig. 2. Preflagellate with nucleus showing a karyosome.
Figs. 3-5. Stages in the division of a preflagellate.
Fig- 3- Preflagellate with blepharoplast dividing.
Fig. 4. Form with blepharoplast and nucleus dividing.
Fig. 5. Separation of the daughter organisms.
Figs. 6-9. Show the development of a flagellum and assumption
of the flagellate form.
Figs. 10-16. A series of flagellates showing variation in size and
appearance.
Figs. 11, 13. Show flagellates containing chromatoid granules.
Fig. 17. An aggregation rosette of flagellates of different ages,
attached by their flagella to a piece of debris.
Figs. 18-22. Stages in the division of flagellates.
Fig. 18. Parasite showing constriction of blepharoplast.
Fig. 19. Parasite with flagellum splitting.
Fig. 20. Form with flagella and blepharoplast separate and
nucleus dividing.
Fig. 21. Commencement of division of the general body
cytoplasm.
Fig. 22. Daughter organisms almost separate.
Figs. 23-36. Illustrate post-flagellate stages obtained from larvae
and pupae.
Figs. 23-25. Transitional forms between flagellate and post-
flagellate stages, as obtained from the haemocoel of the
larva.
Fig. 26. Similar transitional flagellate from the hind gut of a
larva.
Figs. 27-31. Post-flagellates from pupae.
Figs. 32-36. Post-flagellates from the hind gut of a larva.
621
THE CULTURE OF BABESIA
(PIROPLASMA) CAN IS IN VITRO
BY
J. G. THOMSON, M.A., M.B., Ch.B. (Edin.)
(CLINICAL PATHOLOGICAL ASSISTANT, LIVERPOOL SCHOOL OF TROPICAL MEDICINE, AND
PATHOLOGIST, ROYAL SOUTHERN HOSPITAL, LIVERPOOL)
AND
H. B. FANTHAM, D.Sc. (Lond.), B.A. (Cantab.)
(LECTURER ON PARASITOLOGY, LIVERPOOL SCHOOL OF TROPICAL MEDICINE)
(.Received for publication io December , 1913)
Plate XLII
CONTENTS
Introduction .
Technique .
Progress or the Cultures .
Morphology of the Cultural Forms .
Division of Parasites in Vitro .
Successful Inoculation of the Animal Host from a 41 hours’ Culture
Summary .
References .
Explanation of Plate .
page
621
622
622
624
626
629
629
630
632
INTRODUCTION
The present paper contains the preliminary results of the
authors 1 investigations on the cultural forms of Babesia canis . The
importance of the investigation of cultural forms of Protozoa does
not need emphasis, and many practical applications are suggested,
but in this paper we confine ourselves to some remarks on the
morphology and life-history of the Sporozodn as seen in the culture
tube. The method of C. C. Bass (1912) has been followed,
without any addition or modification, though the parasite is not so
easily cultivated as the Plasmodia of man.
622
TECHNIQUE
Cultivation succeeded in two out of four attempts. In each
successful case the infected animal, whose blood was used, was a
puppy about three months old. The blood used in the first culture
was taken from the heart on the fifth day after inoculation, in the
second case on the fourth day after inoculation.
Ten c.c. of heart blood, drawn with aseptic precautions, was
mixed with i/io c.c. of a 50 per cent, aqueous solution of
Merck’s glucose. The blood was carefully and gently
defibrinated by means of a rod, and the clot was removed. It was
noticed that the amount of clot removed was much in excess of that
taken from a similar quantity of human malarial blood. The
defibrinated blood was distributed into smaller tubes, placing about
one inch of liquid in each tube. No centrifugalisation was
necessary. The tubes were incubated at 37 0 C. The corpuscles
settled to the bottom in a short time, leaving a layer of serum above.
Marked haemolysis was seen in all the cultures attempted. It
was found advantageous to take the blood of the puppy before the
crisis, that is, before too many parasites were present in the
peripheral or heart blood.
No sodium citrate nor ascitic fluid was added to the cultures,
as was done by Ziemann (1913), nor citrate and saline as used by
Toyoda (1913).
PROGRESS OF THE CULTURES
We will set forth in some detail the progress of events in our
first CULTURE, which was more successful than the second, in that
more divisions occurred in it.
In the original heart blood, before inoculation, the parasites were
not very numerous, pairs or singles being found in the infected
red blood corpuscle (PI. XLII, fig. 1). Only one group of four
parasites was seen in any corpuscle.* The Babesia were chiefly
pyriform in shape, with small compact nuclei, and the secondary
* Graham-Smith (1905) found that the infected red blood corpuscles containing more
than four Babesia cants constituted less than 0*3 ° 0 of.thc total. He counted 22,589 infected
corpuscles from peripheral and heart blood of dogs. Corpuscles containing one and two
parasites formed 96 4% of the total. ( Journ. Hygiene, V. p. 252)
loose mass of chromatin was not well marked. Phagocytosis was
evident, one large mononuclear leucocyte had ingested.four, infected
corpuscles (PL XLII, fig. 2).
After 7 hours large numbers of intracorpuscular parasites
occurred in clumps, especially at the margins of the smears. The
infected corpuscles usually contained four parasites each (PI. XLII,
fig- 3 )> a f ew contained eight merozoites (PI. XLII, fig. 4). Hence
the original parasites had divided once. The piroplasms, usually
pyriform, now showed clearly the secondary loose mass of
chromatin. Examples of division by budding and chromatin
forking (PI. XLII, fig. 5), as described by Nuttall and Graham-
Smith (1907) from dog’s blood, were seen.
After 15 hours numerous infected corpuscles contained four
parasites, some corpuscles contained eight, while a very few showed
sixteen merozoites (PI. XLII, fig. 7).
At 24 hours there was an increase in the number of infected
corpuscles containing eight parasites, as well as in those containing
sixteen (PL XLII, fig. 8). Corpuscles containing four piroplasms
were also present. Rounded dividing forms, exhibiting chromatinic
forking were found. There was thus evidence of another division
beginning.
After 30 hours some parasites were seen to be degenerating.
Corpuscles containing eight living piroplasms were numerous.
The parasites seemed to have grown larger.
After 48 hours numerous corpuscles were found to contain sixteen
merozoites (PL XLII, fig. 9). One cluster of 29 was found. Three
of the parasites in this group each showed two chromatinic dots, and
were probably about to divide, so that the cluster represented
32 daughter forms derived from one parent Babesia. The host
corpuscle had burst, but its remains could be distinguished.
At 60 hours the parasites were few and were smaller in size.
Two groups of eight and one of four were noticed in one smear.
These parasites had circular chromatin masses. No others were
seen on this smear. Most of the cultural piroplasms were now
dead, and at 68 hours none was found.
Three divisions had occurred in the cultures, from twos, through
fours to eights and sixteens in infected red blood corpuscles.
Heart blood of the dog, kept as control, and incubated at 37 0 C.,
624
showed a few somewhat shrunken parasites after 24 hours, and no
parasites after.
In our SECOND CULTURE only pairs or single pyriform parasites
were found in the heart blood before incubation. This culture
progressed more slowly than the former.
After 6 hours' incubation there was more variety in the form of
the parasites, some being larger and round and some amoeboid.
At 7 hours a very few parasites were showing the commence¬
ment of gemmation.
After 8* hours some free pyriforms were found and an intra-
corpuscular group of four. The nuclei of these contained small
dots (karyosomes) and a little loose chromatin.
At 16 hours fours were more numerous, and distinct chromatinic
budding was seen in several specimens.
After i8£ hours there were numerous groups of four pyriform
parasites, a few eights and one group of twelve. The loose
chromatin was well marked. There was evidence of various types
of division. Some degenerating forms were now noticed.
At 23! hours corpuscles were seen containing one, two and four
parasites.
At 24^ hours groups of four merozoites were fairly common.
At 30 hours many fours were present and some groups of eight.
At 41 hours groups of fours were still found, and | c.c. of the
culture, containing most of the corpuscles, was inoculated into a
young puppy. The puppy developed piroplasmosis and succumbed
five days later. The remains of the culture were examined at about
66 hours and no live parasites were seen.
There is thus evidence that only two divisions occurred in this
culture. It certainly did not grow so well nor so rapidly as the
first. We are unable to explain the cause of this difference in the
progress of the two cultures, but we may remark that the strain in
the second case was of a more chronic character.
MORPHOLOGY OF THE CULTURAL FORMS
The piroplasms were examined fresh, and after fixation and
staining. Smears were made from time to time and fixed by the
wet and by the dry methods, and stained by Romanowsky, Giemsa
625
and haematin stains. Bouin’s fluid was used in some cases as a
fixative.
In this paper we do not propose to deal exhaustively with the
morphology of the cultural forms of Babesia canis , but to record
only the more important findings.
The parasites exhibited marked variation in shape. Pyriform,
amoeboid, round and oval types were seen (Fig. i, A—D). The
method of gemmation and chromatinic forking was observed, as
first described by Nuttall and Graham-Smith from the blood of the
dog. There was also evidence of binary fission.
The pyriform piroplasms (Fig. I, A—B) usually exhibited a
distinct nucleus, as a dot of chromatin, often surrounded by a clear
achromatic halo. Such a nucleus is of the karyosomatic type, the
chromatinic dot representing the karyosome. Such a structure has
Fig. i. Various forms of Babesia canis in culture.
A, B. Two pyriform parasites showing variation in nuclear position. Loose chromatin
also present.
C. Amoeboid form. D. Rounded form.
been recorded in Babesia canis by Schuberg and Reichenow (1912).
The presence of a very thin nuclear membrane is sometimes
suggested, but at other times such a nuclear membrane is certainly
not well marked. A secondary mass of loose chromatin, of a
reticulate or ‘woolly* character is also seen, as described from blood
by Nuttall and Graham-Smith and by Christophers (1907) in Babesia
canis , and by one of us (Fantham, 1907) in Babesia bovis . The
secondary mass of chromatin was well seen in 7-hour cultures. In
some pyriform parasites a very small dot of chromatin was observed
(Fig. 1, B), the so-called blepharoplast of Schaudinn and Liihe;
6 z 6
but this punctiform chromatinic mass is not comparable with the
blepharoplast of a flagellate.
Pyriform Babesia in cultures of 7 hours’ duration had the nucleus
usually near the pointed end (Fig. 1, A), while after 24 hours’
culture it was sometimes seen to be near the rounded or blunt end
of the parasite (Fig. 1, B). Thus the position of the nucleus may
vary in different specimens.
Some amoeboid forms with fine pseudopodia were present,
especially in 7 hour cultures (Fig. 1, C). Such parasites were also
found, motile, on examining fresh preparations.
Rounded forms often showed signs of division by the method
of gemmation.
Fig. 2. Infected corpuscle containing four parasites. The upper one of the four is oval, and
is Leishmania-like ; the lower three show masses of loose chromatin.
In one case, in a 15 hours' culture, a remarkable Leishmania-like
oval form was seen, with nucleus and blepharoplast (Fig. 2, oval
parasite).
DIVISION OF PARASITES IN VITRO
Many examples were found of the mode of gemmation with
chromatinic forking (Figs. 3, 4), now made a diagnostic character
of the genus Babesia. Round piroplasms protrude two small buds
symmetrically arranged to one side (Fig. 3, C), which buds
contain chromatinic cores connected with the main nucleus (Fig. 3,
627
D—G). In some cases the buds are almost entirely composed of
chromatin at first, and seem as if they are about to separate from
the parent. The buds then grow at the expense of the rounded
Fig. 3. Babesia showing division by gemmation, accompanied by chromatin budding
and forking.
A. Parasite with arcuate chromatin.
B. Somewhat amoeboid parasite with chromatin fork.
C. Shows small, symmetrical cytoplasmic buds and chromatin processes entering buds.
D. Parasite with larger buds; main chromatin not distinguishable, but chromatin cores
well marked.
E. Larger buds shown. Well marked main chromatin mass and processes.
F. Parasite showing commencement of fission of main chromatin mass.
G. Form showing typical Y shaped chromatin bifurcation.
portion of the parent parasite, and the linear processes of chromatin
give rise to the loose mass of secondary chromatin seen in the
daughter pyriform piroplasms. Early stages of chromatinic
628
budding and forking were seen while the parasite was still round
or somewhat amoeboid (Fig. 3, B) in contour. Various stages of
division may be observed concurrently in several parasites in one
corpuscle (Fig. 4). Certain cases were noticed in which the
chromatinic forking did not assume the typical Y form (Fig. 3, G),
but was arcuate in character (Fig. 3, A). Sometimes it is difficult
(vide Fig. 3, D) to observe the main chromatin mass from which
the forks arise. It is unnecessary to enter into a lengthy description
of this mode of division, as it has already been described by various
I k.. 4. Infected corpuscle showing parasites in various stages of division.
authors from the blood of dogs. The present is the first record of
its occurrence in cultures, and we also succeeded in seeing the
method in operation in living parasites taken from cultures.
Christophers (1907) considers that, beside the mode described
above, there is also division by direct binary fission. Nuttall and
Graham-Smith have recorded such direct division in the case of
rounded blood forms of Babesia canis. We agree with the above-
mentioned workers, as parasites of various shapes are often seen
with two and even four principal chromatinic masses (Fig. 5, A—B).
We have also seen parasites whose nucleus exhibited a form of
promitosis. Further, we have observed long, somewhat sausage¬
shaped parasites, containing two chromatin masses, and others
629
indented in the middle, apparently dividing into two pyriforms, as
figured by Christophers (1907, p. 21).
Fig. 5. Two parasite* showing direct division.
A has two nuclei, B has four chromatin masses.
The free forms of the piroplasms, seen in cultures, which were
attempting to enter fresh blood corpuscles, were invariably of the
pyriform type, and were endeavouring to enter by the blunt or
rounded ends.
SUCCESSFUL INOCULATION OF THE ANIMAL HOST FROM A
41 HOURS’ CULTURE
A young puppy was inoculated successfully with about £ c.c.
of a 41 hours* culture of Babesia cams . The inoculation was
performed intraperitonea lly. The puppy showed numerous
parasites on the early morning of the fifth day after inoculation,
and died on the evening of the same day.
Unfortunately, the removal of a series of portions of the culture
for examination, has hitherto left us with insufficient material to
attempt sub-cultures. We have, however, no doubt that such sub¬
cultures would be successful, as was shown by Ziemann (1913).
SUMMARY
1. We have succeeded in cultivating Babesia (Piroplasma)
cants in two out of four attempts, following the method of Bass,
using blood and glucose, and incubating at 37 0 C.
2. In one of these cultures, starting with heart blood containing
corpuscles infected with one, two or, exceptionally, four piro-
plasmata, we succeeded in obtaining a maximum of 32 merozoites
in a corpuscle.
630
3. Various types of Babesia were seen in these cultures, namely,
pyriform, amoeboid, rounded and oval parasites. Division of
rounded forms was observed, following the method of gemmation
with chromatinic forking. There was evidence, in stained
specimens, of direct binary fission.
4. Haemolysis occurred in all the culture tubes.
5. A puppy was successfully inoculated from a 41 hours’
culture and succumbed to piroplasmosis on the fifth day.
6. Babesia cants is not so easily cultivated by Bass’s method
as the malarial parasites of man.
REFERENCES
Further references will be found at the end* of the memoir* cited.
Bass, C. C., and Johns, F. M. (1912}. The Cultivation of Malarial Plasmodia (Plasmodium
vivax and P. falciparum) in vitro. Journ. Exper. Med., Vol. XVI, pp. 567-579.
Christophers, S. R. (1907). Piroplasma cants and its Life-cycle in the Tick. Sci. Memoir*
Govt. India, No. 29. 82 pp. Three plates.
Fantham, H. B. (1907). The Chromatin Masses of Piroplasma bigemittum (Babesia bovis ), the
parasite of Texas Cattle Fever. Quart. Joum. Microsc. Sci., Vol. LI, pp. 297-324.
One plate.
Nlttall, G. H. F., and Graham-Smith, G. S. (1907). Canine Piroplasmosis VI. Joum.
Hygiene, Vol. VII, pp. 232-272. Two plates.
Schuberg, A., and Reichenow, E. (1912). Uber Bau und Vermehrung von Babesia cants im
Blute dcs Hundes. Arbeit, a. d. Kaiscrl. Gcsundheitsamte, Bd. 28, pp. 415-434. One
plate.
Thomson, J. G., Thomson, D.. and Fantham, H. B. (1913). The Cultivation of one genera¬
tion of Benign Tertian Malarial Parasites (Plasmodium vivax) in vitro, bv Bass’s method.
Annals Trop. Med. and Parasitol., VII. pp. 153-164. One plate.
Toyoda, H. (1913). Ziichtungsversuche mit Babesia cant's nach der Bassschen Methode.
Centralbl. f. Bakt.. Abt. I, Orig., Bd. 72, pp. 76-81. One plate.
Ziemann, H. (1913). Uber die Kultur der Malariaparasitcn und der Piroplasmcn (Piroplasma
canis) in vitro. Archiv f. Schiffs-u. Tropen-Hvgiene, Bd. 17, pp. 361-391. Two plates.
Also in Tran*. Soc. Trop. Med. and Hyg., Vol. VI, pp. 220-227.
632
EXPLANATION OF PLATE XLII
Fig. 1. Microphotograph showing pyriform Babesia cants from
heart blood of puppy before incubation.
Fig. 2. Shows phagocytosis of four infected red blood
corpuscles. From heart blood before incubation.
Fig. 3. Microphotograph showing infected corpuscles containing
two and four Babesia. Seven hours* culture.
Fig. 4. Division rosette of eight merozoites from seven hours’
culture. Some show loose chromatin.
Fig. 5. Two parasites in process of division by the method of
gemmation and chromatin forking. Seven hours’
culture.
Fig. 6. Shows mononuclear leucocytosis in piroplasmosis,
together with groups of parasites, from a 15 hours’
culture.
Fig. 7. Groups of eight and four parasites from a 15 hours’
culture.
Fig. 8. Group of sixteen merozoites from a 24 hours’ culture.
Fig. 9. Group of sixteen merozoites from a 48 hours’ culture,
showing increase in size.
, tr j s Tr,p. Mci. & Parasitol., Vol. VII
PLATE XLII
i
C. Thomson ami li. B. Fantham. photo.
C. Tinling < 5 ° Co.. Ltd., Imp.
INDEX
INDEX
PACE
Index of Atmtofcs. iii
General Index . iii
Index of Species new to Science . viii
INDEX OF AUTHORS
pace
Balfour, A. .. 113
Barratt, J. O. Wakelin . 367
Blacklock, B. 101
Blacklock, B.; and Stephens, J. W. W. 303
Blacklock, B.; and Yorke, W. ...563, 603
Carter, H. F. 581
Chalmers, A. J.; and O’Farrell, W. R. 525
Chalmers, A. J.; and Stirling, A. D.541
Christophers, S. R. 45
Darling, S. T. 321
Fantham, H. B.; and Porter, A. 569, 609
Fantham, H. B.; and Stephens,
J. W. W. 27
Fantham, H- B.; and Thomson, J. G. 621
Fantham, H. B.; Thomson, J. G.;
and Thomson, D. 153
Kinghorn, A.; Yorke, W.; and
Lloyd, LI. 183
Lloyd, LI.; Kinghorn, A.; and
Yorke, W. 183
Lloyd, LI.; and Wallace, A. F. 299
Macfie, J. W. Scott.1, 339, 359
Marsden, P. H. 335
Mayer, T. F. G. 41
Newstead, R. 331
O’Farrell, W. R. 545
O’Farrell, W. R.; and Chalmers,
A. J. 5 2 5
: FACE
| Porter, A.; and Fantham, H. B. 569, 609
. Rogers, W. 363
Roes, G. A. Park... 371
Scott, H. Harold. 165
Seldelin, Harald . 377
Stephens, J. W. W. 479
Stephens, J. W. W. 5 and Black¬
lock, B.... 303
Stephens, J. W. W.; and Fantham,
H. B. 27
Stirling, A. D.; and Chalmers,
A. J. S+J
Theobald, Fred. V. 591
Thomson, David. 125
Thomson, D.; Fantham, H. B.;
and Thomson, J. G. 153
Thomson, D.; and Thomson, J. G.... 509
Thomson, J. G.; and Fantham, H. B. 621
Thomson, J. G.; and Thomson, D.... 509
Thomson, J. G.; Thomson, D.;
and Fantham, H. B. 153
Todd, John L... 309
Wallace, A. F. 301
Wallace, A. F.; and Lloyd, LI. 299
Yorke, W.; and Blacklock, B. ...563,603
Yorke, W.; Lloyd, LI.; and
Kinghorn, A. 183
GENERAL INDEX
PACE
Acdmorfhus quinquefunctata, nov. sp. 598
Ankylostomiasis, prevention in the
Panama Canal Zone . 137
Anofhelinat, classification and phylo-
geny of . 81
„ Colour marking other
variable characters of 45
„ Geographical distribu¬
tion of . 84
„ Grouping of species ... 87
FACE
Babesia cants , cultivation in vitro ... 621
„ „ morphology in cultures 624
Balfour, A. Trypanosomiasis in the
Sudan and notes on tsetse fly traps
and immune breed of cattle. 113
BankstneUa , Theobald, mosquitos of
the genuS . 581
Barratt, J. 0 . W. Recent experi¬
mental research bearing upon black-
water fever . 367
PACE
Blacklock, B. Posterior nuclear forms
of 7 . rhodesiense in rats . ioi
Blacklock, B., and Stephens, J. W. VV.
Non-identity of 7 . brucei with the
trypanosome of the same name from
the Uganda ox ( 7 . ugandae , sp. nov.)
303
Blacklock,
B., and Yorke, W. Fry -
fanosoma
vivax in rabbits .
563
Blacklock,
B., and Yorke, W. The
probable
identity of 7 . congolense
(Broden) and 7 . nanum (Laveran)
603
Blackwater fever, Correlation between
malaria and black-
water statistics...
499
„ Effect of period of
residence.
488
>>
,, Influence of malaria
486
tt
„ Malarial parasites...
479
tt
,, Pigmented leuco-
cytes .
485
tt
„ Post-mortem exami-
nations .
485
t>
„ Recent experimental
research bearing
upon .
367
tt
,, Relationship to
species of malarial
parasite .
486
tt
„ Seasonal prevalence
492
tt
,, Second attacks.
504
tt
„ Statistical .
479
British Guiana, Sanitation in .
1 43
Carter, H. F. Mosquitos of the genera
Banksinelldy Theobald, and Facnio-
rhynchus , Arribalzaga . 581
Cattle breed immune to trypanoso¬
miasis .119, 124
Cerebro-spinal meningitis, fulminat¬
ing, in Jamaica .*. 165
Chalmers, A. J., and O’Farrcll, \V. R.
The Trichonocardiases . 525
Chalmers, A. J., and Stirling, A. D.
Epidemic Trichonocardiasis . 541
Christophers, S. R. Colour marking
and other variable characters of
Anophelinae . 45
Chrysoconops nociurnus, now sp. ...... 593
Crithidia hyalommae, hereditary infec¬
tion. with, of
Hyalommn
aegyptium ... 545
pace
Crithidia hyalommae} Morphology ... 549
Culicelsa centropunctata , nov. sp. 599
Culicidae from the Sudan . 591
Darling, S. T. Identification of the
pathogenic entamoeba of Panama... 321
Diplococcus jamaieensis , sp. nov.465
Causing vomi¬
ting sickness
in Jamaica ... 465
Domestic Animals, Trypanosomiasis in 1
Dysentery, prevention in the Panama
Canal Zone. 138
Entamoeba , feeding and inoculation
experiments . 326
„ Identification of the
pathogenic, of Panama 321
Fantham, H. B., and Porter, A. The
pathogenicity of Xosema apis to
insects other than hive bees. 569
Fantham, H. B., and Porter, A.
Hcrpetomonas stratiomyiae , n.sp., a
flagellate parasite of the flies, :
Stratiomyia chameleon and S.
potamida , with remarks on the
biology of the hosts . 609
Fantham, H. B., and Stephens,
J. W. W. Measurements of 7 .
rhodesiense and T. gambiense . 27
Fantham, H. B., and Thomson, J. G.
The culture of Babesia (Piroplasma )
canis in vitro . 621
Fantham, H. B., Thomson, J. G.,
and Thomson, D. Cultivation of
Plasmodium vhax in vitro. 153
Filaria loa , note on a case . 363
Fulminating cerebro-spinal meningitis
in Jamaica . 165
Glossina in N. Nigeria . 7
,, austeniin German East Africa 331
„ morsitans , breeding places of 289
,, Development of T.
rhodesiense in ... 273
,. ,, Exper i mental
transmission of
T. rhodesiense ... 187
,. pupation period
influenced by
temperature ... 287
7 . rhodesiense in 215
,. Trypanosomes in
wild . 239
V
PAGE
Glossina sever ini, sp. n. 331
,, „ in the Congo Free
State . 331
Heptaphlebomyia kingii , nov.sp. 601
Hereditary infection, with special
reference to its occurrence in
Hyalomma aegyptium infected with
Crithidia hyalommae . 545
Herpetomonas stratiomyiae, nov. sp. ... 609
„ „ in larva of
Stratiomyia 612
„ in pupa of
Stratiomyia 615
„ „ in imago of
Stratiomyia 616
„ mode of
infection... 617
Hyalomma aegyptium , hereditary in¬
fection with Crithidia hyalommae. .. 545
Ipomoea , root of, from Rhodesia. 335
Isigwebedhla, a fictitious native disease 371
Jamaica, Fulminating cerebro-spinal
meningitis in . 165
„ ' Stegomyia fas data in . 467
„ Vomiting sickness in . 377
„ Yellow fever in . 467
Kinghorn, A., Yorke, W., and Lloyd,
LI:’ ‘ Final report, Luangwa Sleeping
Sickness Commission . 183
Kingia maculoabdominalis , nov. sp. ... 597
Lloyd, LL, Kinghorn, A., and Yorke.
W. Final report, Luangwa Sleeping
Sickness Commission . 183
Lloyd, LI., and Wallace, A. F.
Experiment to ascertain whether
Tabanids transmit trypanosomes in
nature . 299
Loa loa , A case of . 363
Luangwa Sleeping Sickness Com¬
mission, final report . 183
Macfie, J. W. S. Trypanosomiasis of
domestic animals in N. Nigeria.. 1
Macfie, J. W. S. Morphology of the
trypanosome ( T . nigeriense , n.sp.)
from a case of sleeping sickness from
PAGE
Malarial Parasites, Cultivation of.. 510
, „ ,, Cultivation of P.
vivax in vitro... 153
„ Growth and sporu-
lation in the cul¬
ture tube and in
the human host 509
Marsden, P. H. Root of an Ipomoea
from Rhodesia . 335
Mayer, T. F. G. Mosquito-proof and
storm-proof house . 41
Meningitis, Fulminating . cerebro¬
spinal, in Jamaica . 165
Micrococcus castellanii , sp. nov. 531
„ „ In trichono-
cardiasis ... 528
„ nigrescens , in trichono-
cardi^pis . 528
Mosquito-proof and storm-proof house
for tropics . 41
Mosquitos of the genera Banksinella
and Taeniorhynchus . 581
Mucidus nigerrimus, nov. sp... 591
Muscidae in N. Rhodesia . 296
Newstead, R. New tsetse-fly from
the Congo Free State; and the
occurrence of G. austeni in German
East Africa . 331
Nocardia tenuis in trichonocardiasis... 528
N. Nigeria, Glossina in. 7
„ Trypanosomiasis in domes¬
tic animals in . 1
Nosema apis , pathogenicity to insects
other than hive bees 569
,, „ in various Hymenoptera 570
„ „ in various Lepidoptera 572
„ „ in various Diptera. 575
O’Farrell, W. R. Hereditary in¬
fection, with special reference to its
occurrence in Hyalomma aegyptium
infected with Crithidia hyalommae 545
O’Farrell, W. R., and Chalmers, A. J.
The Trichonocardiases . 525
Ornithodorus moubata in N. Rhodesia 293
Ek'et, S. Nigeria. 339
Macfie, J. W. S. The development
of a human trypanosome in the gut
of Stomoxys nigra . 359
Malaria, prevention in the Panama
'Canal Zone. 130
„ ,, transmission ex¬
periment with
7 . rhodesiense 301
Panama Canal Zone, Sanitation in ... 125
„ Pathogenic Entamoeba of ... 321
Piroplasma canis , cultivation of. 621
Y1
PAGE
Plague, prevention in the Panama
Canal Zone . 137
PUsmodium falciparum , Morphology in
culture. 513
„ „ Morphology in
the human
host . 516
., vivax, Cultivation in vitro 153
„ Morphology in cul¬
ture . 518
„ „ „ Morphology in the
human host ... 519
Porter, A., and Fantham, H. B. The
pathogenicity of Nosema apis to
insects other than hive bees. 569
Porter, A., and Fantham, H. B.
Herpetomonas stratiomyiae, , n.sp., a
flagellate parasite pf the flies, S,
chameleon and S. potamida , with
remarks on the biology of the hosts 609
Pupipara in N. Rhodesia . 297
Reedomyia sudanensis , nov. sp. 595
Rhodesia, Northern, blood-sucking
insects and
ticks in the
Luangwa
Valley 293
„ „ Meteorological
observations 189,190
Rogers, W. A note on a case of Loa
loa . 363
Ross, G. A. P. A fictitious native
disease {Isigwebedhla) . 371
Sanitation in the Panama Canal Zone 125
Scott, H. H. Fulminating cerebro¬
spinal meningitis in Jamaica. 165
Seidelin, H. ‘ Vomiting Sickness * in
Jamaica . 377
Sleeping Sickness Commission, Final
report of the
Luangwa . 183
„ ,. Distribution of, due
to 7 . rhodesiense 185
„ „ in Eket, S. Nigeria 339
„ „ symptomatology ... 186
„ „ Sex and age of
Africans suffering
from . 309 1
Southern Nigeria, sleeping sickness due
to 7 . nigeriense, n.sp. 339
PACE
Sudan, cattle breed immune to try¬
panosomiasis .119, 124
„ New Culicidae from the. 591
„ Trypanosomiasis in . 113
Stegomyia fasciata in Jamaica . 467
Stephens, J. W. W. Studies in black-
water fever . 479
Stephens, J. W. W., and Blacklock, B.
Non-identity of 7 . brucei with the
trypanosome of the same name from
the Uganda ox ( 7 . Uganda e, sp. nov.) 303
Stephens, J. W. W., and Fantham,
H. B. Measurements of 7 . rhodes¬
iense and 7 . gambiense . 27
Stirling, A. D., and Chalmers, A. J.
Epidemic Trichonocardiasis . 541
Stomoxys possibly transmitting try¬
panosomiasis . 8
„ nigra , trypanosome in gut of 359
Stratiomyia chameleon , infected with
Herpetomonas
stratiomyiae 609
„ „ Biology of. 610
„ potamida , infected with
Herpetomonas
stratiomyiae 609
„ „ Biology of. 610
Stratiomys , vide Stratiomyia ....609
Tabanidae in N. Rhodesia . 293
Tab anus , trypanosome transmission
experiment . 299
7 aeniorhynchus, Theobald, mosquitos
of the genus. 585
Theobald, F. V. New Culicidae from
the Sudan . 591
Thomson, D. Sanitation in the
Panama Canal Zone, Trinidad and
British Guiana. 125
Thomson, D., Fantham, H. B., and
Thomson, J. G. Cultivation of
Plasmodium vivax in vitro. 153
Thomson, D., and Thomson, J. G.
Growth and sporulation of the
benign and malignant tertian
malarial parasites in the culture tube
and in the human host. $$9
Thomson, J. G., and Fantham, H. B.
The culture of Babesia (Piroplasmd)
canis in vitro. 62 1
mi
PAGE
Thomson, J. G., and Thomson, D.
Growth and sporulation of the
benign and malignant tertian
malarial parasites in the culture tube
and in the human host. 509
Thomson, J. G., Thomson, D., and
Fantham, H. B. Cultivation of
Plasmodium vivax in vitro. 153
Todd, J. L. Sex and age of Africans
suffering from Trypanosomiasis. 309
Trichonocardiasis . 525
„ epidemic . 541
Trinidad, Sanitation in. 141
Trypanosomes in the gut of Stomoxys
. 359
„ in wild G. morsitans ... 239
„ of game and domestic
stock . 227
„ Transmission experi¬
ment with Tabanids 299
Trypanosomiasis, Cattle breed immune
to.119, 124
,, Human, vide Sleep¬
ing Sickness.
„ Immunity. 10
„ In cattle, seasonal
distribution .7, 8
„ Incubation and
duration of, due
to 7 . rhodesiense 103
„ In the Lado(Western
Mongalla, Sudan) 113
„ Of domestic animals
in N. Nigeria. 1
„ Prophylaxis . 9
„ Transmitted possibly
by Stomoxys . 8
„ Treatment . 9
„ Tsetse fly traps 118, 124
7 . brucei in domestic animals.2, 3, 4
„ Non-identity with the try¬
panosome of the same
name from the Uganda
ox ( 7 . Ugandae, sp. nov.) 303
„ Posterior nuclear forms. 3
7 . congolense from donkey . 122
„ Probable identity of, and
7 . nanum . 603
7 . gambiense , Measurements of . 27 j
7 . ignotum , sp. nov. 263 ]
„ Diagnosis. 266
PACE
7 . ignotum In wild G. morsitans , 240, *242
„ Measurements . 264
„ Morphology . 263
„ Pathogenicity. 265
„ Transmission . 269
7 . multi forme, sp. nov. 254
„ Diagnosis . 257
„ In game in N. Rhodesia 233
„ Measurements .255, 259
„ Morphology. 254
,, Pathogenicity . 257
„ Transmission . 261
7 . nanum In domestic animals, 2, 3, 7, 23$
„ In game in N. Rhodesia, 231,
233 * 234
„ Measurements . 249
„ Morphology . 248
„ Pathogenicity . 248
„ Probable identity of, and
7 . congolense . 603
„ Transmission. 251
7 . nigeriense , n. sp. 356
„ Animal reactions . 345
„ Measurements.... .348
„ Morphology . 343
7 . pecorum from bull. 122
In domestic animals, 2, 3,7, 235
In g*me in N. Rhodesia, 233,
234
„ In wild G. morsitans , 240, 243
„ Measurements. 252
„ Morphology . 251
„ Pathogenicity . 253
„ Transmission . 254
7 . rhodesiense . 245
„ Development in G. mor¬
sitans . 273
„ Development in G. mor¬
sitans influenced by
,, meteorological con¬
ditions . 204
„ Distribution of . 185
„ In domestic animals ... 23$
In game in N. Rhodesia, 233,
2 34
In G. morsitans in nature
215, 240, 242
Measurements of ... 27, 218
Pathogenicity of. 223
Posterior nuclear forms
in rats . 101
PACE
7 . rhodesiense Reservoir of. 214
„ Transmission experi¬
ments with G. morsi-
tans . 187
,, Transmission experi¬
ment with 0. moubata 301
7 . theileri in domestic animals . 3, 7
T.tragelaphi, sp. nov. 269
„ Diagnosis. 269
„ In game in N. Rhodesia 234
„ Morphology . 269
. „ Transmission . 270
7 . Ugandae, sp. nov. 308
„ In domestic animals . 3, 4
„ Posterior nuclear forms ... 3
7 . vivax In domestic animals, 2, 3, 6, 235
In game in N. Rhodesia, 231, 233,
234
„ In rabbits . 563
„ Measurements of . 246
„ Morphology of. 245
„ Pathogenicity of. 247
„ Transmission . 247
Trypanosoma , sp. (? montgomeryi), diag¬
nosis . 263
„ sp. (? montgomeryi) in
domestic animals ... 235
„ sp. (? montgomeryi), mor¬
phology. 261
PACE
Trypanosoma, sp. (? montgomeryi).
measurements. 262
„ sp. (? montgomeryi),
pathogenicity . 261
„ sp. (? montgomeryi),
transmission . 263
Typhoid, prevention in the Panama
Canal Zone. 138
Vomiting sickness in Jamaica .. 377
Wallace, A. F. Attempt to transmit
7 . rhodesiense by means of O. mou¬
bata . 301
Wallace, A. F., and Lloyd, LI. Ex¬
periment to ascertain whether
Tabanids transmit trypanosomes in
nature . 299
Yellow fever in Jamaica . 467
„ ,, Prevention in the Panama
Canal Zone . 136
Yorke, W., and Blacklock, B. Try¬
panosoma vivax in rabbits 563
Yorke, W., and Blacklock, B. The
probable identity of 7 . congolense
(Broden) and 7 . nanum (Laveran)... 603
Yorke, W., Lloyd, LI., and Kinghorn,
A. Final report, Luangwa sleeping
sickness commission . 183
INDEX OF SPECIES NEW TO SCIENCE
PACE PAGE
Aedimorphus quinquepunctata . 598
Chrysoconops nocturnus . 593
Culicelsa centropunctata . 599
Diplococcus jamaiccnsis . 465
Glossina sever ini . 331
Heptaphlcbomyia kingii . 601
Herpetomonas stratiomyiae . 609
Kingia maculoabdominalis . 597
i Micrococcus castellanii . 533
I Mucidus nigerrimus . 591
Reedomyia sudanensis . 595
Trypanosoma ignotum . 263
„ multiforme . 254
„ nigeriense . 356
„ tragclaphi . . 269
,, ugandae . 308
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