(navigation image)
Home American Libraries | Canadian Libraries | Universal Library | Community Texts | Project Gutenberg | Children's Library | Biodiversity Heritage Library | Additional Collections
Search: Advanced Search
Anonymous User (login or join us)
Upload
See other formats

Full text of "The bacteriological world : a monthly illustrated magazine devoted to the study of micro-organisms and specific maladies"




Illliii IP il 

ifff; 



PI^BW 











»««» 







t 




V 



Digitized by the Internet Archive 

in 2012 with funding from 

Open Knowledge Commons and the National Endowment for the Humanities 



http://archive.org/details/bacteriologicalw1 1 891 colu 




Of 

HI 

t- 
co 

CO 

O 




x, 
o 
o 
*: 

cc. 




rru 




fllD 




r\ 



*u 



GICA 



u 




A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies, 

Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological World, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to by Dr. 
T. J. Turner, Mexico, Missouri, to whom remittances should be made. However, 
all moneys sent to the Bacteriological World Co., Columbia, Mo., will be received 
and promptly acknowledged. 

Authors of original articles published in this magazine will be entitled to twenty 
copies of the number containing it, provided, that they request the same in writing 
when sending the communication. Reprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. 

Discontinuation :— At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first of next January. 



collaborators: 



Paul Gibier, M. D., New York. 
Prop. W. T. Belpield, M. D., Chicago, 111. 
Prop. A. W. McAlester, A. M., M. D., 
Missouri University Med. Dep't. 



Prof. L. Bremer, M. D., St. Louis, Mo. 
J. W. Stickler, M. D.. Orange, N. J. 
Prop. Paul Schweitzer. Ph. D., 

Missouri University. 



U. S. ft, 



1,1. 



EDITORIAL. 



With hearty good wishes for a Happy New Year, we hum- 
bly present to the medical public our new-born babe the 
Bacteriological World. It was born, we hope, with suffi- 
cient natural vigor to insure its existence, provided that it 
receives proper nursing from its parents and close friends. But 
it cannot grow strong, broad, and become a useful and powerful 
factor unless it receives from the outside a sufficiency of good 
stimulating substantial food, i. e., scientific and financial aid. 
Our first plea, then, in behalf of our creature, is for scientific 
communications and subscriptions. 



2 THE BACTERIOLOGICAL WORLD. 

The Bacteriological World is a new departure in pro- 
fessional and scientific journalism on this continent, and is the 
child of necessity. Though realizing our shortcomings, we 
have dared to bring forth this creature because the medical 
and scientific fraternities, and individuals, on this side of the 
ocean, feel the need of something of the kind daily, and have 
waited in vain for years for its birth. Since the discovery 
of the role of micro-organisms in disease, it has become neces- 
sary for the medical and veterinary practitioners to inform 
themselves on the subject. Indeed, it is now indispensable to 
know bacteriology as well as anatomy, and to be as well 
acquainted with the physiology of microbes as the physiology 
of man. None can understand true pathology as is now clearly 
demonstrated without at least a fair knowledge of pathogenic 
bacteriology, and consequently none can practice medicine and 
surgery as safely, as successfully, as conscientiously and as fairly 
towards his patients as he who is familiar with the new science. 
It matters not what unbelievers think of this proposition ; it 
matters not whether they ridicule the idea or not ; I say with 
all due respect, that, at this date of the nineteenth century, a 
medical man, young or old, who refuses to admit the factor of 
living germs as causative agents in disease is, to his own detri- 
ment, far behind the times in medical science, and seemingly 
shows a disposition to cling to old ideas whether right or wrong. 
For no man, I dare say, can study the history, life and prop- 
erties of bacteria, without realizing that there is not more 
mystery about the parasitic bacteria than there is about the 
parasitic tapeworm. So far as the existence of the two is con- 
cerned we can prove the one more easily because the organism 
is visible to the naked eye, and the other, — the bacteria, — unfor- 
tunately requires to be magnified. 

Physicians, veterinarians, students in all branches of 
medicine are, therefore, in need of knowledge in bacteriology ; 
and, since it is so new a science, comparatively speaking, they 
are bound to disappointment, if they desire to advance and 
depend exclusively on text-books appearing from time to time, 
for, though these works are all good and necessary, every day 
brings, us something new and important from the various 



THE BACTERIOLOGICAL WORLD. 3 

pathological and bacteriological laboratories of the world. 
Note, for instance, the recent wonderful discovery of Koch ! 

It is as reporter of everything useful, important and 
needed in medical bacteriology, that we come to all those inter- 
ested in the form of this humble magazine, and propose to 
bring to every home, office and college the discoveries of the 
investigators of the universe. We offer, in these humble pages, 
to supply early the American continent with every available 
new point relating to microbes and useful in medicine, that 
may be brought to light by any of the experimentalists and 
observers in Germany, Austria, France, Italy, England, Amer- 
ica, and all other portions of the scientific world. We propose 
to bring to the busy practitioner, who can scarcely leave home 
for special investigations, every new observation, new experi- 
ment in specific diseases, etc., and besides, we respectfully 
offer him an illustrated plain course in bacteriology at a cost 
incomparably lower than he could possibly obtain it by any 
other means. All medical men have not the advantage of a 
second schooling or of laboratory pursuits abroad, but still all 
are equally in need of the information that is thus obtained 
by the few. Will they avail themselves of our respectful offer 
to come to their aid in our humble way through the Bacteri- 
ological World ? We have the sanguine hope that they will 
and thereby sustain and nourish our infant journal. Without 
the support of the medical fraternities we cannot prosper and 
perhaps must die ; even the personal sacrifices that we had to 
make in the shape of financial investment of all our little pos- 
sessions will be lost unless we meet with a hearty welcome, and 
the warm sympathy of our friends, and the friends of medical 
progress and science. 

In conclusion, we very respectfully dedicate this : our first 
issue of 5,000 copies of the Bacteriological World to our 
many kind friends in the medical profession, and to the friends 
of science and medicine. May they peel generously dis- 
posed towards our new-born child and tender it the 
milk of life — subscription. 

To our new friends in the field of journalism we respect- 
fully bow and ask for a little corner in their glorious and 
honored field of labor. 



4 THE BACTERIOLOGICAL WORLD. 

KOCH'S TREATMENT OF TUBERCULOSIS. 

From all accounts to date, the eminent physicians, who 
have followed the experimental treatment of tuberculosis by 
Koch's injection, all agree that it is a success in cases where no 
serious injury exists in any important organ of the system. If a 
success, it is without doubt the most beneficial discovery that 
ever was made in any branch of medicine. Several other 
glorious discoveries have been made in science which benefited 
the world in many respects, and among medical discoverers we 
may point with heartfelt thankfulness to a Harvey, a Jenner, 
a Pasteur and other luminaries, but none of the past or present 
can bring such gladness to almost every household in the 
world — among the rick, the poor, and even the savages, as this 
man Koch if his lymph proves successful. 

Tuberculosis is so widely disseminated, so fatal and so far- 
reaching in its ill results upon the nations, that a remedy to 
cure it in its incipiency must be hailed with joy by all men and 
all nations. Yes, it is a matter of congratulation that 
science " n'a pas de patrie " — that science has no home but the 
univei'se at large, so that a useful discovery, be it made in 
France, Germany, England, Russia, America, or even in 
"darkest Africa," if clearly given to the world, for the good 
of humanity, is received with gratitude by all mankind. 

The achievement of Koch has something in it which 
makes a true man's heart feel reverent. His was not an acci- 
dent, nor was it the result of accidental observations and occur- 
rences, but it was the fruit of medical and bacteriological 
knowledge applied with accuracy and perseverance, guided by 
intelligence and honesty, and sustained by a most energetic 
will power. He discovered the germ of tuberculosis about 
eight years ago, and in defiance of most determined opposition 
and most serious objections continued his labors to this moment 
and with the results known. 

We understand that Dr. Koch means to publish the 
methods by which he prepares his fluid. We expected no less 
from so great a man, but we fear that a too early explanation of 
the processes may cause accidents, and perhaps delay the good 
that must come to all nations if only they will have the 



THE BACTERIOLOGICAL WORLD. 5 

patience to wait. In the first place, though a man may be a 
good, scientific and learned medical practitioner, he may not be 
versed sufficiently in laboratory technique to prepare this fluid, 
and the failures, disasters and deaths that might follow impure 
injections must redound to the injury of the whole new treat- 
ment. Indeed, if it takes so much care to produce safe, active 
bovine vaccine that as yet only few experts succeed satisfacto- 
rily, how much more difficult and how much more accurate 
must be the technique of producing this fluid in a field so new 
and still so dark as bacteriology ? It must be apparent to all 
medical men, that only comparatively few can safely and suc- 
cessfully produce the delicate material, and no man is justifi- 
able in injecting in his fellow men a substance other than from 
an absolutely reliable source, proved to be such by the character 
and knowledge of the producer. It seems to us, therefore, 
that the process should be parted with only after a safe trial at 
Berlin, and then, too, entrusted first to men capable and fit to 
carry on the production with absolute correctness. 

We sincerely hope, in justice to the famous and honored 
discoverer, and in justice to the human race, that the excite- 
ment, existing now in medical circles and the world over, will 
not cause a premature exposition of the secret. While most 
reputable physicians flock to Berlin to witness the treatment 
for the benefit of humanity, it will be found, as in all other 
things, that the only aim of some is to turn the discovery into 
dollars, and they at once would become purely business manu- 
facturers. We find similar concerns now among medicine pro- 
ducers. 

It is to be hoped that the wildness now prevailing will 
soon subside, and that Prof. Koch may thereby find competent 
men to whom he will demonstrate his methods of procedure, 
and thus put every mortal within reach of his blessed treat- 
ment. 

PASTEUR— KOCH. 

Our portraits of the celebrities, Louis Pasteur and Eobert 
Koch, will doubtless gratify all who may see them. No two 
scientists are more popular universally than these two grand 
geniuses, and no two men living to-day, and perhaps of the 



6 THE BACTERIOLOGICAL WORLD. 

past ages, have rendered as great service as they have to 
medicine and the human race generally. 

Pasteur, probably the greatest chemist of the century, 
discovered and led the way, from a chemist's standpoint, to the 
wonderful fountain of knowledge in the etiology and prevention 
of disease, and has since, himself, caused the application of his 
marvelous discoveries for the benefit of mankind, particularly 
in his treatment of hydrophobia. Koch, working from a botan- 
ical basis, broadened the path, illuminated the darkest passages 
and led a whole army of investigators through successful 
excursions and explorations in pathology. His admirable 
achievements in technical bacteriology, and his great discoveries 
concerning tuberculosis and other affections, have made for 
him a name which will outlive the youngest of the present 
human race. Yea, the names of Pasteur and Koch will live to 
the end of time, and will be forever cherished by all generations 
as two of the greatest benefactors that ever blessed the world. 
These two scientists being the first and foremost bacteriologists, 
it is eminently appropriate that we should here present their 
pictures to oar readers. Doubtless at this particular time the 
face of Koch will prove specially interesting, because his recent 
discovery of a successful treatment for consumption now 
occupies the minds of the medical men and the masses. 

May Pasteur and Koch be spared long years of prosperity, 
happiness, and of scientific labor, is the sincere wish of the 
Bacteriological World. 



The subscription for the Bacteriological World for 
one year is S3. 00 : single copies, 30 cents. But we offer here 
three months' trial subscription ( in advance ) for 75 cents. 
Considering the cost of publishing illustrated magazines, this is 
as low as we can afford until at least we secure 5,000 subscribers. 
We urge all medical men, of both human and veterinary medi- 
cine, and all students interested, to subscribe at once if they 
wish the course in bacteriology complete, for we have issued 
only 5,000 copies of this first number, have but few left, and can- 
not afford another edition unless we get a very large demand. 
Hence, to secure a complete course illustrated on bacteriology 



THE BACTERIOLOGICAL WORLD. 7 

(that may be bound at the end of the year ), including all the 
new discoveries that may occur in the meantime, one should 
subscribe noio. We may have nothing of this first edition left 
in a short time and we can afford to print in the future only 
enough to supply subscribers. Send $3.00 to the Bacterio- 
logical World Publishing Company, Columbia, Mo., or to 
Dr. T. J. Turner, Mexico, Mo. See business announcement 
with table of contents on the second page of advertisements in 
the last pages of this issue. 



COLLABORATORS AND CONTRIBUTORS. 

Among the foremost medical men who have commended 
and spoken kindly of our enterprise, may be mentioned : 
Doctors Mitchell Prudden, of New York ; Paul Gibier, Pas- 
teur's Institute, New York ; J. W. Stickler, of Orange, New 
Jersey; W. T. Belfield, of Chicago, 111.; W. C. Vaughan, of 
Ann Arbor, Mich.; I. N. Love, Editor of the Medical Mirror, 
St. Louis ; Professors McAlester and Moss, Medical Depart- 
ment of the Missouri State University, And some of these 
highly reputed men of science have promised to favor our 
readers with articles. Their names are a guaranty of the 
character of the scientific and practical work that they may do 
for our subscribers. 

In the next issue we shall present probably a longer list 
of honored and trusted names as regular collaborators and con- 
tributors. Among them we hope to have specialists in vege- 
table micology, as we mean to present comparative studies in 
this department of bacteriology. 



THE BACTERIOLOGICAL WORLD, 

LESSONS ON BACTERIOLOGY. 

{Copyrighted.) 



BY PAUL PAQUIN, M. D., V. M.. 

Missouri State University. 



PREFACE. 

To understand fairly the so-called ''germ diseases " it is 
essential to know to some extent the life history of the microbes 
in general and of some of them in particular. Unfortunately, 
most physicians cannot study this satisfactorily because they 
are so situated, that they must depend mostly on abstract writ- 
ings for light. There are good books on Bacteriology — indeed 
some of them are admirable and all have their place and utility, 
and yet scarcely any one combines in a satisfactory practical 
manner the natural history of micro-organisms with the tech- 
nical branches of the new science. From a practicing physi- 
cian's standpoint, there is, in my judgment, something lacking 
in these excellent works to make them of as much practical 
value as they might be in their hands, and to all interested in 
medicine. Some are very voluminous and so deep as to be use- 
ful chiefly to students with leisure, or to experts ; others are too 
exclusively technical. In thinking about the matter it occurred 
to me that a series of plain lessons on the question of micro- 
organisms, chiefly from a medical standpoint, would serye a 
good purpose among many of those desirous of acquiring some 
practical information on the matter. Accordingly I have 
concluded to publish such lessons in the Bacteriological 
World. Practically the same course is taught by the author 
at the Missouri State University. 

In writing these lessons I shall follow the highest authori- 
ties. 

INTRODUCTION. 

The word bacteria, in reference to the so-called " germs of 
disease," is rather restrictive, because it implies that among 
micro-organisms, the class known as bacteria is alone capable 
of causing disease. It is thought now that such is not exactly 
the case, there are pathogenic micro-organisms that belong 



THE BACTERIOLOGICAL WORLD. 9 

seemingly to other classes of beings. The word microbe seems 
more suitable as a general term. 

However that may be it is absolutely proven, that many 
diseases are due to living forms of the bacteria group. In these 
maladies the exceedingly minute microscopic beings are para- 
sites, and, generally speaking, some authors think that micro- 
parasitic organisms would express better their significance than 
." disease germs," though there are some very small parasites 
more highly organized than bacteria. 

Bacteria live almost everywhere in the air, water, soil, on 
and in living bodies, etc., and still comparatively few of them 
are injurious ; these are the microbes which happen to find 
congenial soil in living tissues and fluids. 

The microbic or bacterial diseases then are after all para- 
sitic affections. So far as their etiological classification is con- 
cerned, they might be included under the ordinary heading of 
parasitic maladies though there is a vast difference as to the 
size and organization of the parasites, for all such agents have 
properties in common : that is, that whether large or small, 
visible or invisible with the naked eye, they live at the expense 
of other organisms and transform portions of the latter's 
bodies for their own nourishment. 

The bacteria have been termed Schizomycetes by JSTageli ; 
they are unicellular organisms, and their mode of digestion and 
life is such that the transformations which they cause in the 
nourishing material harboring them is wonderful and great, 
incomparably greater, indeed, than that of higher organisms. 

It becomes necessary then, in order to understand microbic 
affections, to study the physiological conditions, and somewhat 
the biological chemistry, of these infinitely little creatures in a 
general way and compare with that of more complex creatures. 
These two subjects are in themselves important, deep and long, 
and consequently, in an abbreviated course, could not be thor- 
oughly treated. But we can and will touch the points essential 
for a fair knowledge of pathological bacteriology. 

BRIEF HISTORICAL NOTE. 

The leptothrix of the saliva, and vibriones of the foeces 
were described by Leuwenhoek over two hundred years ago as 



10 THE BACTERIOLOGICAL WORLD. 

animalcules, but it is scarcely more than twenty-five years that 
the study of " germs " in relation to disease took a decided 
step in advance. The natural history of micro-organisms had 
been studied in the interval by Ehrenberg, Dujardin, Robin, 
Hallier, Cohn and others, but it was not until Pasteur identified 
the phenomenon of fermentation- with the life and growth of 
microscopic forms that the scientific world opened its eyes as 
from a dream. In Germany the renowned Koch diligently 
labored on a botanical basis, and invented accurate methods of 
bacteria investigation ; in England Lister astounded the 
medical world by his successful applications of Pasteur's discov- 
eries to surgery. The living nature of ferments being known, 
every country of the globe launched into the new, fascinating 
field of natural history. And now there is scarcely a civilized 
nation without some illustrious bacteriologist. But, with all 
that, our knowledge of the natural history of micro-organisms 
is still very limited. The bacteria are believed to be vegetables, 
but the place of some of the finer monads seems yet a mooted 
question. Generally speaking many micro-organisms apparently 
lie in the obscure, indefinite border-land between the animal 
and the vegetable kingdoms. Science may eventually draw a 
line and place all of them where they belong. 

In these lessons I desire to bring forth, among other things, 
the intimate relations of some microbes with the tissues and 
fluids of the body, and their actual actions in producing disease 
with references to plant affections. Therefore, it matters 
not so much from a practical standpoint to what kingdom the 
parasites belong, if we can understand their properties, and the 
results of their existence. 

LESSON FIRST.— PART I. 

GENERAL IDEA OF FERMENTATION AND LIFE. 

Pasteur in his investigations in organic chemistry discov- 
ered and proved the living nature of ferments as I have already 
mentioned. The w T orld was forced to admit that such phe- 
nomena as are witnessed in the boiling-like action of the leaven 
of bread, the fermentation of beer, wine, cider, etc., in the 



THE BACTEEIOLOGICAL WOELD. 11 

formation of vinegar, are due to the growth of minute organ- 
isms in the media in fermentation. Eealizing then that the 
great changes, that occur in such fermentations, really are trans- 
formations of certain matter into other substances, he reasoned 
(after long investigations), that possibly the minute foreign 
bodies found in certain diseases (such as had been noticed in 
charbon by Devaine) were also some kind of ferments, and 
that the changes or lesions found in the tissues, blood, etc., 
were nothing other than alterations and transformations, a kind 
of fermentation due to the growth and sustenance of these 
individuals at the expense of their hosts. This preconceived 
idea has been amply supported by facts since proven. Parasites 
of the microbic order are ferments of some kind or other, and 
their relation to their hosts may be compared with the relation 
existing between the microscopic vinegar plant and the alcohol 
which it (the plant) transforms into vinegar : or the relation 
is the same as that of the putrefaction ferments and the dead 
bodies that they decompose. In all of these cases there is alter- 
ation and transformation more or less complex and more or less 
complete. The only striking difference is, that germs living at 
the expense of live tissues, such as septic microbes in a wound, 
encounter the natural resistance of the organized living cells of 
the body that they have invaded, whilst a ferment in dead 
matter encounters no resistance from an opposing life. 

The parasitic germs ( or ferments, or microbes, or bacteria, 
or schizomycetes if you please) cause disease, therefore, simply 
because they find in animal bodies (and even in plants) ele- 
ments that they can transform and utilize for their own nour- 
ishment. They break up more or less living material, and by 
that means manage to exist whilst causing thereby very natur- 
ally more or less disturbance in the invaded organism. The 
invasion is resisted in a degree by the constituting ( and new 
ceils generated) in the system attacked, and there ensues a fight 
for supremacy and existence. In this struggle many microbes 
lose their lives and many body cells perish. If the microbes event- 
ually conquer fully the disease may terminate in death ; if the 
cells (aided perhaps by chemical properties) of the afflicted organ- 
ism conquer, whether taking medical assistance into account or 
not, there may be enough vitality left to rebuild the. constitution. 



12 THE BACTERIOLOGICAL "WORLD. 

It is a sublime sight to watch day by day, with a microscope, 
certain parasites devouring blood for instance, and to note the 
reaction of leucocytes and other cells with destructive effects on 
the microbes. It is a wonderful struggle ! A grand battle ! 
The cells of the body are our soldiers, and their defense of the 
fort against the armies of parasitic germs is admirable and 
sublime. 

Now what constitutes a ferment? What distinguishes a 
microscopic being of the order of ferments from other organ- 
isms in their mode of existence ? To grasp the differences and 
relations, one must bring to the mind a general idea of life. 
It is essential, indeed, to glance at the processes of nutrition, 
growth and even generation of organisms of a superior order. 

If a seed is sown in a fertile soil — one well fertilized, sav 
with thoroughly decomposed barn manure — it will swell by the 
action of moisture, give rise to a plant which will grow more or 
less luxuriously to maturity. JSTow, how is it possible for that 
little seed to grow into a plant thousands and perhaps millions 
of times larger than itself and of a texture totally different in 
appearance, color^ density, etc. ? The seed sends forth its first 
leaves ; these leaves by the light fill with chlorophyl, thereby 
becoming capable of taking carbon,, and the new being gradually 
manufactures living material at the expense of air *and mineral 
substances in the water of the soil. There is a transforma- 
tion of matter ; from dead substances life is extracted. The 
plant is a laboratory which utilizes the forces received from solar 
heat to bring about more or less complex and strange combina- 
tions of gaseous or soluble elements of the globe, resulting in 
tissues and organs as we find them in the scale of vegetable 
beings. This is the result of their nourishment. 

Animals live directly from plants, or from other animals 
having lived directly or indirectly at the expense of vegetables. 
So that we may safely say, that the source of life in animals 
( man included of course ) is, after all, the same as that of 
plants. A being in the animal world springs also from a seed. 
It is at first nourished at the expense of the mother who gets 
her food directly or indirectly from plants. The foods of 
animals are transformed too, and from dead matter serve to 
make new living matter. But once formed those organized 



THE BACTERIOLOGICAL WORLD. 13 

tissues of plants and animals are soluble no more. Formed of 
gaseous or soluble elements, there has been erected a wonderful 
structure that in nature^ is proof against the action of water. 
The fibrous tissues of a tree and its tough bark, the fibrous 
and muscular tissues of an animal and its tough skin are alike 
insoluble. And all that, mind again, has come directly or indi- 
rectly from elements of the air and water. Suppose then, that 
the atmosphere and the water did not receive by some means a 
new supply of these necessary elements for the formation of 
living organisms, what must be the inevitable result ? Uni- 
versal failure of life. The world must get back a supply of the 
elements used and necessary to form bodies or life cannot be. 
The air once devoid of these limited organizable elements 
necessary to form tissues in plants and animals, there can be no 
more growth or generation of living things. JSTow what does 
return these organizable elements to the universe ? Ferments ! 
Germs ! Microbes ! 

These invisible mites are indeed the " counterweights nec- 
essary " to the existence of large plants and animals. They 
cause the putrefaction of dead bodies and their transformation 
into gaseous soluble matter to be utilized by new plants and 
animals. All that lives must die, and after death the carcasses, 
the debris, are decomposed, fermented, and the air and water are 
thereby replenished with the organizable elements necessary for 
the existence of species. So it is that from the dead rise the 
living, and that death is necessary to perpetuate the races. 

We readily understand, then, that bacterial existence is of 
absolute necessity, though many microbes happen to cause 
much injury and disease because they find material fit for their 
nourishment in other living beings, such as larger plants and 
animals. 

To prove that plant life, and hence animal life, depend 
largely on the continual transformations of dead matter of the 
globe into organizable elements, as above explained, it is only 
necessary to sow seeds in sterilized earth. Such soil being 
deprived of the fertilization due to microbes, such as nitre 
etc., etc., barely causes the seeds to sprout ; they show a puny 
yellowish stem, and then perish ; or perhaps the seeds them- 
selves perish at the outset of their germination. A sterilized 



14 THE BACTERIOLOGICAL WORLD. 

soil cannot nourish plants. This was demonstrated by Duclaux. 
Whether a maple tree, a man or a microbe, life is in each in a 
measure dependent on the other two classes ; and if one king- 
dom could be dispensed with it is the animal kingdom. It is 
plain also, that life in each, class can be sustained only by the 
power that individuals have of transforming matter in various 
ways for their nourishment. 

low, we must go a step further in this general outline of 
life, if we would understand the actual processes of transforma- 
tion in each living thing, including germs. 

In the first place, a large plant absorbs from the soil and 
air soluble matter that it uses, as was before stated, to manu- 
facture its tissues, etc. It finds the food substances already 
prepared, and only has to make new substances out of them. 
Generally speaking it does not have first to digest or decompose 
its food by special secretions. And yet there is a time in a 
complex plants existence when such a secretion occurs to pre- 
pare food for a special purpose. A sugar beet for instance 
stores in its roots, during vegetation, a certain amount of sugar 
which, though constantly in a watery media, never dissolves 
until the period of flowering. Why does it not dissolve until 
then ? Because the sugar is of the insoluble kind in water 
(stored for the construction of the flower); but then a diastase is 
secreted by the living tissues of the beet itself — a diastase which 
has the property of transforming insoluble sugar into soluble 
sugar. During this gradual transformation or preparation of 
sugar the plant appropriates that which it needs to construct 
the flower. Note the point. A diastase is secreted that 
changes the sugar into a soluble assimilable kind. 

Now consider a man's digestion. Among many other 
things that he eats and which are transformed by the various 
secretions of the digestive apparatus is starch. This substance 
reaches the intestines and is yet unfit to be assimilated, but the 
pancreas secretes a diastase that transforms the starch into 
soluble sugar which then can serve as food for the economy. 

The life of microbes depends on the same thing, but being 
unicellular organisms instead of complex and highly organized 
bodies, they do not take in the food and digest it within their 
own systems. They digest it outside if it happens to be proper 



THE BACTERIOLOGICAL WORLD. 15 

for their nourishment and unfit for their assimilation. They 
secrete digestive diastases which prepare the food. Thus it is 
that a piece of flesh is putrefied and even liquefied to some 
extent by germs to serve as their nourishment. The process 
of putrefaction is merely a feeding phenomenon. 

But, whilst nutrition goes on and diastases of various kinds 
are secreted to prepare the food, complex living individuals 
reject also waste materials in the form of gas, liquids and 
solids. It is only necessary to remember the urinary excretion 
and the gaseous exhalations of various beings to appreciate 
this. Well, the same thing is true of microscopic beings, they 
secrete, excrete. And now these gaseous, liquid and solid 
materials wrought by the feeding processes of microbes are set 
free more or less in the universe ; they come in contact with 
the chemical elements of the air and water (and even with 
each other ), and thus chemical combinations bring forth a 
variety of more or less important products. It is thus that we 
get vinegar, alcohol, nitre, ammonia, ptomaines, leucomaines 
and various other chemical substances and combinations, either 
harmless, beneficial or poisonous in various degrees. 

Now it may be asked, how can such small beings as 
bacteria or ferments suffice to the task of transforming ail dead 
matter, etc., etc. ? You may form an independent opinion on 
that point as we progress, but it seems to be chiefly because of 
their wonderfully rapid propagation, and the property of each 
to decompose matter so much more in excess of their own 
weight, than is possible in larger and more highly organized 
individuals. For example : A seed in germination consumes 
daily about 2-100 part of the weight of the nourishment within 
its hull ; a dog destroys daily an average of 1-25 of its weight in 
food ; a man about 1-50 ; fowls about 1-4 of their weight. If 
we compare that with microscopic beings we find that some of 
these, like the vinegar plant ( mycoderma-aceti ) for instance, 
transform about 100 times their weight. The majority have 
still far more il destructive energy." 

It is perhaps well to give here a preliminary idea of the 
rapid reproduction of microbes. These organisms reproduce 
themselves by fission and by formation of spores (seeds). 
Pasteur in studying the vinegar plant actually saw, from one 



16 THE BACTERIOLOGICAL WORLD. 

single cell, such rapid reproduction during two hours ( in a 
poor medium too), that in 24 hours he calculated that more 
than 16,000,000 individuals had been generated. This is 
striking but by no means the most rapid reproduction of bac- 
teria. Comparatively speaking this is indeed slow. 

At this period in our study, we have already an idea of 
what are bacteria and of their role in the world. We know 
now that while acting as scavengers they preside at the fertiliza- 
tion of the soil, and prepare therein, and in the air, food to be 
utilized by plants and by animals ; we know that they are use- 
ful in arts and sciences in the manufacture of fermented prod- 
ucts such as bread, wines, etc.; we know again that some are 
parasitic and live at the expense of other living organisms, and 
thereby cause organic changes termed lesions, with accompany- 
ing reactions and phenomena which are the symptoms. 

A germ disease then is a specific disease, and, I repeat, 
a parasitic disease, and the symptoms are only the expression 
of the struggle within between the invading cells and cells of 
the invaded individual. 

PART II. 

SPONTANEOUS GENERATION.* 

We have had a glimpse of the very interesting role of 
micro-organisms in the world, and we are brought to face with 
the question of "spontaneous generation." Whence do these 
microbes come ? Do the minute living cells rise spontaneously 
in dead matter ? Are they born by a " regressive metamor- 
phosis, by a natural transformation of some organic matter" ? 
Could they possibly be generated by any other means than 
those governed by the normal laws of generation ? 

In olden times this question was answered in the affirma- 
tive by all scientists. No less a great genius than Aristotle 
attributed the generation of eels to the fermentation of the 
slime of rivers ; and the generation of caterpillars to the putre- 
faction of earth or plants under the influence of dew. Virgil, 

*The great majority are informed on this question, and to them this 
portion of my course may seem superfluous ; but a few are still in 
obscurity. 



THE BACTERIOLOGICAL WORLD. 17 

in the fable of Aristee, " recounts the birth of a swarm of bees 
in the entrails of a bull." This may be taken as an indica- 
tion of the beliefs at the time — beliefs which would seem older 
than the Christian era, for the Book of Judges in the Old 
Testament creates bees in the bowels of a lion. 

It were a too long analysis to review the history of the 
eminent supporters of this theory. Suffices it for our object to 
point to the affirmative belief in spontaneous generation by all 
naturalists through all ages until this century. 

Not very many years ago this view was doubted, and the 
French Academy of Sciences offered a prize for the solution 
of the problem. It was solved absolutely and clearly in the 
negative by Pasteur. We know now positively that there is no 
such occurrence as spontaneous generation of living organisms 
no matter how little. The smallest ones have been generated by 
parents. The most minute mass of living individuality had 
procreators. The era of the microscope has led the way to a 
clear insight of the dark abyss, and experiments innumerable 
have, since Pasteur's discovery, verified his assertions. 

It is very easy to satisfy one's mind on this point. If a 
piece of flesh, cooked or otherwise, be exposed, it will putrefy, 
and perhaps maggots will form in the mass. These two occur- 
rences were once both attributed to spontaneous generation at 
the expense of the flesh aided by mysterious air influences » 
But suppose another piece of flesh is heated in a jar, closed only 
by a plug of cotton, through which the air may penetrate, leav- 
ing its germs in the cotton fibers, it will not putrefy, and no 
life can be found in it. The heat in this case has destroyed 
life, and the plug of cotton prevents new ingress of organisms. 
Neither the air nor the meat then have the power to cause the 
generation of the germs. And all organic substances can be 
preserved indefinitely by simply killing ( by this or other pro- 
cesses ) the germs in and on them, and preventing their access 
to them afterwards. 

The germs as all other beings are generated ; we know 
so little about them because our natural optics are insufficient 
to see them. 

The so-called "germ theory" then, is supported by facts 
resting on a solid basis everywhere and in every sense. After 



18 THE BACTERIOLOGICAL WORLD. 

we realize the unicellular microscopic size of the bacteria, 
their rapid propagation by normal processes, their ferment 
properties, we find the question of " disease germs " wholly 
devoid of the mysterious shroud that surrounds it, in the minds 
of those ignorant on the subject. 

In closing this first lesson I would recapitulate briefly, and 
fix in your minds a few cardinal points: 

First. That the words germ, bacteria, microbe, schizomy- 
cetes are used in our present literature almost as synonymous 
terms, but microbe seems preferable to germ or bacteria, and 
schizomycetes is a better scientific term than either. 

Second. That these are unicellular, and assimilate nour- 
ishment, seemingly by absorption in the media in which they 
live, but, 

Third. That they must transform, alter the foods found 
proper, and yet unfit in nature, for their use and appropria- 
tion. 

Fourth. That bacteria living on dead matter encounter 
no living resistance, whilst •those feeding on living tissues, or 
fluids in living tissues, meet the living cells of the body and 
have to combat them. 

Fifth. That the diastases secreted by the various beings, 
whether highly organized or unicellular and microscopic, have 
something in common as to their respective objects, and their 
properties of transforming matter. 

Sixth. That the role of microbes in the world is complex 
and necessary, though some are injurious. They act as scav- 
engers, return to the air and water the organizable elements 
abstracted daily by the vegetables of the globe, and indirectly 
by animals, and indispensable to life. 

Seventh. That the bacteria, that invade living organisms 
which happen to be fit for their nourishment and growth, are 
in a sense parasites just as much as the tapeworm is. 

Eighth. That spontaneous generation of living organisms, 
no matter how little, is a fallacy. 

(Our next lesson, in February, will be illustrated in the text. P. P.) 



THE BACTERIOLOGICAL WORLD. 19 

SURGICAL BACTERIOLOGY. 



BACTERIAL COMPLICATION OF WOUNDS. 



BY A. W. MCALESTER, A. M., M. D., 

Dean, Medical Department, Missouri University. 



(From investigations of Ogston, Rosenbach, Cornil, Babes and others.) 

As the study of pathological micology advances, we realize 
the nature of many hitherto misunderstood complications of 
wounds. Until late years we thought the complications in the 
practice of surgery were scarcely more than two : Septicemia 
and pyemia, and various affections were classed as ordinary 
medical cases, which bacteriology has placed definitely among 
the surgical diseases. Under the heading of surgical affections 
we now find (from the classification of Cornil and Babes): 
vesico-pustules of the dorsal region of the fingers ; paronychia 
(•felon, whitlow); conglomerated perifolliculitis; furunculus 
and anthrax ( boils ) ; erysipelas ; warm abscesses ; phlegmons ; 
lymphangitis ; thrombus '; phlebitis ; metritis and puerperal 
inflammation ; pyemia ; septicemia ; sapremia ; arthritis ; oste- 
omyelitis ; certain forms of endocarditis, myocarditis and 
nephritis ; omphalitis ; hemorrhagic septicemia ; gangrene ; 
gaseous gangrene ; tetanos. 

Some of these, such as erysipelas and furunculus, appear 
often as spontaneous, and. many are the physicians who deny 
their specificity. It is to Ogston and Eosenbach that we owe 
most of our knowledge on the etiology of these surgical affec- 
tions. Their deep investigations have thrown a brilliant light 
on all the complications of wounds of various nature. 

Though the bacterial diseases consecutive to wounds have dif- 
ferent appearances, and present different specific symptoms, they 
nevertheless have something in common. They are all charac- 
terized by inflammation, and in nearly all there is some form 
of suppuration more or less pronounced. Several of them 
may produce the complications known as septicemia, pyemia, 
gangrene, and several may cause embolism. On the other 
hand, the biology of the respective causative agents establishes 
their relation. 



20 THE BACTERIOLOGICAL WORLD. 

The pathogenic microbes, discovered in connection with 
these diseases, nearly all belong to the class of cocci, which 
are disposed in chains or masses. The most common are : 

First. Staphylococcus aureus, found by Pasteur in 
furunculus, and existing in the abscesses of pyemia and of 
osteomyelitis. It was seen also by Oornil and Babes in some 
cases of ulcerated endocarditis, gangrenous pneumonia, puru- 
lent pleurisy, cerebro-spinal meningitis and parenchymatous 
nephretis. Externally it was found in the air, soil, kitchen 
water, on the surface of the skin, under the nails, in soiled 
clothes of healthy children. It is occasionally found associated 
with the parasites of actinomycosis, tuberculosis, glanders and 
other maladies. By inoculation it can by itself produce abscesses. 

Second. Staphylococcus flavescens, resembles much the 
preceding, and has much the same properties. It may be 
distinguished chiefly by the differences in artificial cultures. 

Third. Staphylococcus pyogenes alius which has the prop- 
erties of the preceding, but differs in cultivation. 

Fourth. Streptococcus pyogenes. This organism, which 
appears in the form of chains, presents several species, one of 
which seems to be the actual cause of erysipelas, and another 
the cause of warm abscesses, though, as we have said above, 
other related micro-organisms have been found in such cases 
also. 

Fifth. Streptococcus septicus of pus. It resembles the 
preceding somewhat in shape, but is much more pathogenic in 
pure inoculations. 

Now there is a microbe described by Passet, that justly 
belongs here on account of its properties, though it is placed in 
a group of differently shaped germs. It is the bacillus pyogenes 
fostidus. A strong odor of putrefaction characterizes its 
growth. 

In many complications of wounds, various other germs 
may be found associated with these specific pathogenic forms, 
but they seem accidental visitors. Possibly, too, they work 
injury in company with the others. It is easy to appreciate the 
means of ingress in various surgical cases, such as amputations, 
abdominal operations, obstetrical cases, etc. ; but it is difficult 



THE BACTERIOLOGICAL WORLD. 21 

to explain the entrance of microbes in such diseases as furun- 
culus and some cases of erysipelas. This point is somewhat 
theoretical. The appearance of furunculus is usually in regions 
where the skin is subject to friction, such as the neck by collars, 
the wrists by cuffs. Lowenberg thinks that the bacteria pene- 
trate by the pilo-sebaceous follicules. Whatever may be their 
means of ingress, they may be found in the affected parts even 
though no visible external opening be discovered. G-arre has 
shown, it is written, that the staphylococcus may penetrate 
even through healthy skin. 

The same thing may be said as to the entrance of the 
parasite of erysipelas, in which sometimes no visible wound is 
present. Frequently, though, a slight scratch or abrasion had 
existed and given passage to the organisms, and then healed, 
leaving no trace. 

The local bacterial complications of wounds (such as have 
been mentioned ) may pass off by nature's own efforts or aided 
by medical or surgical means. They are individually specific 
in their nature and somewhat independent in their localization, 
though, as we have seen, related from an etiological standpoint. 
Microbes of the same order, and with properties very similar, 
have independent pathogenic powers resulting in the lesions 
found in each disease locally. But they may be found associ- 
ated and thus complicating still more a wound, and again, as 
stated, they may have the company of germs ordinarily harm- 
less, which find in the diseased tissues fit soil for their growth 
and multiplication, thereby aggravating the primary complica- 
tions. 

Pyemia and septicemia are the results of local bacterial 
complications. At first, there may be only a local abscess, or 
suppuration of some character or other in a wound ; both are 
the result of germ life. But soon fever appears, there is subde- 
lirium, prostration, and various symptoms indicating general 
infection or intoxication of the system. It is partly the result 
of migration of the septic microbes in some important organs, 
perhaps their generalization in the circulation, and it indicates 
besides and chiefly the general poisoning by the toxic products 
generated by them. 



22 THE BACTERIOLOGICAL WORLD, 

The nervous symptoms are conclusive evidences that an 
impression has been produced by a toxic substance. Such 
nervous poisoning may be compared as to its origin with alco- 
holic intoxication — alcohol being the product of a living 
ferment. 

The most important point in the treatment of wounds, 
therefore, is the prevention of access of micro-organisms. With- 
out them there can be no complication such as has been men- 
tioned here, and hence no pyemia, no septicemia. 

There is another form of blood poisoning ; it was named 
Sapremia by Duncan, Ogston and Eosenbach. It is that which 
results from intoxication by products of putrefaction. Occa- 
sionally, germs of putrefaction decompose blood or diseased 
tissues in living individuals, and produce nauseous odors and 
poisonous substances. Such things occur more frequently in 
obstetrical cases. It goes without saying that this compara- 
tively rare form of septic poisoning can be avoided as the 
others mentioned, and by the same means. 

Besides the complications that I have mentioned, there 
are a few others of serious character, that deserve special dis- 
cussion, but my space is too limited to do them justice. 
However, I will mention them. As far back as 1856 Virchow 
and Beckman recognized zooglea in ulcerous endocarditis. 
More recently Mayer, Klebs, Orth and others gave us a good 
description of the bacterial, granulous and ulcerous forms of 
this inflammation. 

In their recent and admirable work on bacteria, Cornil 
and Babes describe pyemic myocarditis, and illustrate the 
presence of cocci in the muscular tissue of the heart. 

In the same work we find descriptions of various bacterial 
nephretis consecutive to wounds or infectious maladies. 

Tetanos is another usually fatal malady now classed among 
the parasitic kind of bacterial order. The germs are found in 
the earth, manure and other places, whence they may pene- 
trate through small or large wounds into the system. 

But enough for this time, hoping to be able later to treat 
each malady individually. 



THE BACTERIOLOGICAL WORLD. 23 

HYGIENIC BACTERIOLOGY, 



VACCINIA, PATHOGENIC BACTERIA AND VACCINATION. 



BY PAUL EVANS, 

Pathological Laboratory of the Mo. Agr'l Expt. Station. 



Since the discovery of so many pathogenic germs in con- 
nection with disease, and their ability to enter the organism 
through the slightest wound ; since we know of the existence 
of so many kinds of septic germs in water, air, etc., particu- 
larly where there is organic matter in decomposition, the medi- 
cal practitioner often fears to inoculate children or others 
with vaccinia. The country is flooded with announcements of 
vaccine propagating farms, and the means at hand for a physi- 
cian to know the responsibility of propagators, and the purity 
of their products are in most cases wofully lacking. 

On the other hand, on the physician's side, the study of 
the diagnostic symptoms of true vaccination free from com- 
plication has been neglected by many, as has been to some 
extent even the method of the minor operation known as vacci- 
nation. 

There is need of improvement until absolute care and 
cleanliness becomes the rule — such aseptic methods, indeed, on 
the part of both vaccine propagators and physicians, as will 
positively exclude all germs except the specific microbe of 
vaccinia itself. From observations and experience it strikes 
me that this is not always the case. 

Germs found in" vaccine. — Dr. Paquin, in studying the 
germs of vaccinia in virus obtained from three different 
places in the United States and one in Europe, found in almost 
every case germs capable of causing complications if they had 
been in sufficient number, whilst in the virus of one source he 
found other pathogenic germs than those of vaccinia in great 
abundance — enough to make it dangerous. In a new operat- 
ing room with limed walls, Paquin made the following tests, 
the details of which nearly all came under my observation : 
Case I. Condition of room was apparently clean, no floating 



24 THE BACTERIOLOGICAL WORLD. 

particles could be seen with the naked eye but no disinfection 
had been practiced. A heifer about six months old was 
shaved closely on the abdomen ; the surface of the shaved skin 
was carefully washed with apparently clean water and soap, 
and then dried with a clean cloth. Adhering to the surface of 
this washed skin were found still a few microbes. Ten scari- 
fications were made and each was rubbed with vaccinia. 
After all the spots had been so inoculated they contained air 
germs in a much greater quantity than had been found on the 
skin, these had fallen during the operation and adhered to the 
raw surfaces. Of course many must have been rubbed in with 
the vaccinia. The result of this inoculation was that two of the 
ten scarifications developed a diffused inflammation and swell- 
ing far more extensive than in a typical pustule of vaccinia. 
In studying the virus pressed out gently therefrom, it was 
found to contain less comparatively of the vaccinia cocci, far 
more serum than in a typical pustule, and a large quantity of 
the cocci known as staphylococcus aureus. These were culti- 
vated artificially and well identified. Only a very few of these 
foreign germs were observed beneath the scab of some of the 
pustules which presented a normal condition, but none in the 
oozing lymph from them. 

Inoculations with cultures of these pyogenic cocci in rab- 
bits produced diffuse local inflammations occasionally followed 
by suppuration. 

A negro boy working in the laboratory was vaccinated on 
the arm by the writer with virus from such an abnormal 
pustule, and containing these cocci in a large quantity, and a 
good pustule was the result, but, besides this, a serious painful 
inflammation and hard swelling occurred extending to the elbow 
in one direction and to the shoulder in the other, involving 
the axillary glands considerably. Cultures made with virus 
from the pustule of this negro produced unmistakable pus 
cocci which were inoculable and pathogenic in rabbits. 

Now, whilst vaccine apparently pure sometimes causes 
marked and serious symptoms, it is probable that most of such 
cases as just described are complications due to the introduction 
of germs that were on the skin, the lancet, in the air, the water 
used to wash, or, possibly, in the virus itself. 



THE BACTERIOLOGICAL 'WORLD. 25 

Case No. 2. The operating room was disinfected by 
burning sulphur bricks during the night with closed doors. In 
the morning a calf was laid on the table, shaved, washed 
with soap and ordinary soft water, then thoroughly rinsed 
with freshly boiled water, and the surface wiped with a towel 
that had been heated 212° F. in a- dry oven. The lancet 
used had been sterilized in a gas blaze, the hands of the oper- 
ator washed in a bi-chloride of mercury solution 1-2000, and 
the sleeves were covered from the wrists to the elbows with 
clean protectors. 

Ten scarifications were made and inoculated with neat, 
dear ivory points from Martin, of Boston. The result was 
ten typical pustules, free from extraordinary diffuse swelling or 
inflammation. The virus secreted thereby contained no other 
pathogenic microbe than vaccinia ; pure cultures failed to 
produce dangerous germs. 

Case No. 3 was a case of inoculation in the same condition 
as No. 2, but ivory points this time slightly tinged with blood 
were used. The result were good pustules secreting pure tymph 
with vaccinia cocci and no other germs. 

My observations, since, of Moss'' vaccinations at The Paquin 
Vaccine Laboratory, where aseptic methods are carefully fol- 
lowed, confirm me in the views expressed here as to the cause 
of various serious symptoms in vaccination. 

I should have mentioned that none of the calves were pro- 
tected by covering, and that, as a consequence, the surface scabs 
were in all cases more or less impregnated with external germs, 
but in lifting the healthy scabs, as stated, and wiping the raw 
surfaces underneath with a sterilized cloth, these typical pustules 
secreted pure virus, whilst the unhealthy-appearing pustules 
secreted virus contaminated, as explained in case 1. 

These incidental studies, though scarcely sufficient and 
satisfactory, point to the possible danger from impure virus, 
and to a moral that a conscientious physician should always 
apply, to-wit : Know the source of your vaccine before apply- 
ing it. It is not sufficient to know that it is bovine virus, or 
to know that in a given case this or that virus has given 
remarkably good results or produced " strikingly good pustules 
and swelling"; we should know further the nature of the 



26 THE BACTERIOLOGICAL WOELD. 

pustule, and the qualification and character of the vaccine 
producer. 

In order to produce safe bovine vaccine ( and no other 
kind should be used ), it is evident from the above that several 
conditions must be fulfilled from an aseptic standpoint. 
First, the stable should be kept scrupulously clean and so 
built as to allow effective washing of floors, and disinfection of 
all of its parts and of the air ; it should be also perfectly 
drained and ventilated. Second, the calves should be thor- 
oughly examined by a competent veterinarian, and their bodies 
should be cleaned daily and well brushed. Third, the operat- 
ing room should be so constructed as to be as near air-tight as 
possible to favor as perfect sterilization of the air and as per- 
fect cleansing and sterilization of the walls, floors, etc., as may 
be had ; such disinfection should be repeated every evening. 
Fourth, the operator's hands and clothes should be free from 
dust and dirt of any kind. Fifth, the water used should be 
sterilized by boiling ; the towels sterilized by dry heat ; the 
lancets sterilized by passing them in a blaze ( allowing them to 
cool afterwards ). Sixth, inoculate calves only with virus 
known to be pure. Seventh, a vaccinated animal should be 
protected by some special covering, to prevent contamination of 
the inoculated spots by excrements, urine and bedding, with- 
out interference with the development of the pustules. The 
bedding should be heavy, clean and constantly renewed. 
Eighth, in collecting the crop, the same aseptic precautions 
ought to be taken with the operating room, the operator and 
instruments, as stated already ; and the ivory points, receiving 
and drying vessels, ought all to be free from living germs. In 
dipping points in such a cleanly room, with hands perfectly 
clean, the vaccine must be pure if it comes from a good healthy 
pustule from which kind only the virus should be gathered. 
Ninth, physicians, to be on the safe side in vaccinating, should 
cleanse the skin perfectly with freshly boiled water in prefer- 
ence to any other ( never with an antiseptic drug which may 
kill the vaccine germs afterwards ), sterilize their lancet, then 
make a couple of good scarifications until a slight oozing of 
blood ; after wiping this off with a clean cloth, rub in the 
virus of a moistened ivory point heavily charged. 



THE BACTERIOLOGICAL WORLD. 27 

Now, if inoculation so practiced, is allowed to be contam- 
inated by the friction of a soiled shirt or drawers, there may be 
complications yet ; therefore, it is well to protect the scarifi- 
cations with a piece of silk oil cloth applied over them. 

The examination of cattle by a veterinarian before vaccin- 
ation is necessary to exclude all animals showing the least sign 
of disease, such as tuberculosis for instance, which is so com- 
mon in cattle and communicable to man. 

These bacterial complications of vaccination may explain 
some failures in vaccination of people, and of calves used for 
the production of vaccinia. For instance, germs producing 
much serosity in the pustule may cause death of the vaccinia 
cocci by virtue of the bactericide property of serum. Again, 
the possible antagonism between the bovine vaccine microbes 
and other germs in a pustule may result in weakening or 
destroying the virulence or life of the former. The writer has 
known vaccinia sent 1,000 miles to the south in the warm por- 
tion of summer, tried by physicians without success, returned 
after three weeks or more, then tried on calves to take per- 
fectly. May we not conclude that in some such occurrences 
the vaccinator inoculated in the arm or leg, or allowed to pene- 
trate various air germs antagonistic to vaccinia, and that the 
latter suffered thereby ? How often are not unclean arms 
scratched, inoculated and then covered with a soiled raiment ? 



THEORY OF INFECTIOUS DISEASES, CURE, VACCINATION 
AND NATURAL IMMUNITY. 

(Report by Chas. Bouchard to the Academy of Sciences, Paris, France.) 

I can deduct from the experiments made, that I have 
already communicated to the Academy, the conclusions which 
constitute a new systemization of infection and immunity. 

If the infectious agent inoculated falls into an animal 
organism which is a medium very bactericide, it does not 
develop itself; it does not outlive the disease ; if the living 
animal constitutes a media very favorable, the microbe develops 
itself immediately; if the media is moderately bactericide, 
there is in the life of the microbe a first phase of degeneration ; 



28 THE BACTERIOLOGICAL WORLD. 

during a short enough time ( a few quarters of an hour ) its 
multiplication is suspended ; but it lives meanwhile, and, 
through its diastase, adapts to its needs the material of the 
tissue where it has been deposited ; then its interrupted devel- 
opment disappears. 

When the development of the pathogenic agent was 
immediate where it had been preceded by a phase of degen- 
eration, the malady commences. In time they multiply them- 
selves, the microbes secrete chemical substances in larger and 
larger quantities, some of which act on the nervous system, 
causing the change of the circulation or of the modification, 
nervous headache, delirium, coma, convulsions, etc.; all the 
others impressing all the cells of the body, changing their 
nutritive type, and, by their intermediate agent, modifying the 
chemical composition of the humors which can thus become 
bactericides. This last effect is tardy, but it is durable. The 
poisonous substances which impress the nervous system have a 
1 more rapid action, but more temporary. Among these rapid 
and transitory actions is the paralysis of the vaso-dilatator ner- 
vous centers, which renders impossible the exit of the white 
globules outside of the vessels. 

When the number of the microbes becomes such that their 
products of secretion constitute a mass which can no longer be 
ignored, the febrile and poisoning symptoms appear. The 
bactericide state does not exist yet; the phagocytism alone 
could save the menaced organism ; but the phagocytism is 
rendered impossible, because, while secreting the other poison- 
ous substances, the microbe has secreted the material which 
prevents diapedesis. Thus in a media which is still chemically 
favorable to its development, and protected against the most 
important of the cellular reactions, the microbe continues to 
multiply and secrete freely, the intoxication augments, the 
malady grows worse ; death may supervene in this period. 

But, during this time, the materials which change the 
nutrition of the cells were secreted also, but their slow action 
does not manifest itself yet. In a moment they have impressed 
the cells enough in order that their nutritive type be modified, 
in order that the humors be, in consequence, changed chem- 
ically. The bactericide state is a possible effect of this chemical 



THE BACTERIOLOGICAL WORLD. 29 

change. It appears slowly, but as soon as it appears the life of 
the microbes is impressed, their pnllulation slackens or arrests 
itself, their secretions suspend themselves. The material which 
opposes the diapedesis, in particular, comes no more to par- 
alyze the vaso-dilatator nervous center. Then the white globules 
come out of the vessels, and the phagocytism destroys at last 
the microbes already attenuated by the bactericide state. This 
is the cure. 

The cure is the first manifestation of the acquired 
immunity. The bactericide state, produced slowly by the 
passing impregnation of the cells in contact with vaccine 
substances, persists a long time after the elimination of these 
materials. If, in a vaccinated individual, this bactericide state 
is very pronounced, the microbe which produced the first sick- 
ness, introduced by a new inoculation, cannot vegetate ; there 
will be neither a general infection nor a local infection ; the 
immunity is absolute. If the bactericide state is less pro- 
nounced, it does not prevent the life of the microbe, but it 
attenuates it, it diminishes the activity of its secretions ; one of 
these, in particular, becomes incapable of paralyzing, as for- 
merly, the vaso-dilatator center ; the diapedesis is not prevented, 
and the phagocytism arrests and terminates the infection in its 
primitive point. The local lesion has been rendered possible 
and it has prevented the general infection from presenting 
itself. 

The natural immunity does not depend on the bactericide 
state ; it results in a greater resistance than, in certain species 
of animals, the vaso-dilatator center opposes to the material 
with paralyzing properties. The proof is that the diapedesis, 
normally provoked in these animals by certain microbes, 
pathogenic in other species, does not occur if we inject with 
the dose of virus to which they resist, a stronger dose of 
the chemical substance which prevents diapedesis. We notice, 
then, that the diapedesis and, consequently, the phagocytism 
do not take place ; we notice further that general infection 
produces itself. 



30 THE BACTERIOLOGICAL WORLD. 

ORIGINAL ARTICLES. 



ACTINOMYCOSIS. 



( By the Editor.) 

This is a specific disease of much interest in the- United 
States to-day. It exists among cattle, in the West at least, to 
an alarming extent — our live-stock markets witnessing hun- 
dreds of cases monthly. I have seen personally a good number 
of cases in official capacity, and all the "members of the veteri- 
nary sanitary force of the State of Missouri have had more or 
less extensive experience with it. In view of the possible trans- 
missibility to man, it has become a question of national import- 
ance, and one which has been too long neglected by sanitarians. 

The question, involving as it does such a vast industry as 
cattle raising and beef production, is one in which various 
interested parties seem to interpret to suit their respective cases 
without regard to facts and the general good. Indeed, conflicting 
opinions have been given by several reputable veterinarians and 
physicians seemingly without regard to the most recent investi- 
gations and discoveries. From various parts of the country 
have arisen defenders of this and that interest, in Illinois for 
instance, when state and city authorities and the stock-owners 
conflicted this year. If all those who ventured opinions on the 
subject would have searched the history of the malady fully, its 
pathology, causation, dissemination, and the most accurate of 
the old as well as of the new investigations, experiments and 
discoveries, they must have come to the unanimous verdict 
which some of them, in the cases alluded to, have wisely 
reached, i. e. 3 that actinomycosis is a specific, parasitic and, 
therefore, transmissible affection. Whether directly or indi- 
rectly transmitted is not the all-important point, but the fact 
that it is at all transmissible in any way is the interesting 
feature. And when we consider that actinomycosis in man 
and beast is identical, we have still a far more interesting 
subject. 

The writer is at present carrying on experiments in con- 
nection with this disease, and is not prepared just now to speak 



THE BACTERIOLOGICAL WORLD. 31 

fully on all points pertaining to its pathology and its parasite 
from a personal investigation, but he will treat the disease as 
fully as possible in these pages under the circumstances, and 
cite authorities at his command. 

This writing is merely from a medico-scientific standpoint, 
and not in any sense meant to enter into discussion with any of 
the medical men who may have expressed different views. 

Etiology ae"d history, — Actinomycosis is due to a 
vegetable parasite growing in tissue thereby causing chronic 
inflammations of various degrees ; it was named " actinomyces 
bovis" by Bollinger, in 1877 (Germany), after the results of 
Harz's investigations made under the former's direction. The 
name was given because of its radiated form ( aJctin, ray, and 
muJcos, mucous) at that time its filaments had not been dis- 
covered, or had been misinterpreted. These little vegetables 
appear grouped in yellowish granules. But years before that 
Langenbeck { Germany ) and then Lebert ( France ) had given 
the first anatomo-pathological observations of the affection in 
man. Eobin (France ), in 1871, observed identical lesions and 
parasites and misunderstood their nature. This is as far as the 
study was carried in man until after the disease was well 
described in cattle. 

In the meantime Sebastiano Eivolta ( Italy ) found the 
actinomycosis granules in studying sarcoma of cattle, but 
attached no significance to them until 1878, when he again 
investigated new cases and became convinced of their parasitic 
nature. In 1875, Perroncito (Italy ) studied osteo-sarcoma of 
cattle and wrote the nature of the actinomyces, coming to the 
conclusion that it was a cryptogam. 

These investigations stimulated others, and again the 
study of the disease in mankind was undertaken. Israel 
( Germany ) described three cases in man. Ponfick, in 1879, 
identified the similarity of actinomycosis of man and cattle. 
And since then Europe has reported over fifty cases in human 
beings, whilst a dozen or so are recorded here and there in 
America as having been identified. Having so much of it in 
cattle, I think that some cases of actinomycosis in man, like 
cases of glanders, are not recognized ; I knew of one case in 
Southern Missouri which local physicians diagnosed by various 



32 THE BACTERIOLOGICAL WORLD. 

names. The man had probably contracted the disease from an 
ox which was suffering from actinomycosis at the lower 
maxillary ; the ox and the man had the habit of drinking from 
the same trough. The lesions in the man were in one of the 
maxillary articulations. 

Authorities state that in man the disease is always char- 
acterized by two appreciable things : Abscesses and yellowish 
granules in the pus of the abscesses. In cattle it is not always 
so. The parasites growing in the muscular tissue of the tongue 
for instance cause this organ to become as hard as wood, and it 
is free from pus. However, pus cavities of various sizes are 
the most common lesions in cattle also, and in them the yellow 
granules as seen in man are always found though sometimes 
with difficulty. 

Lesions. — I do not intend to describe the pathological 
anatomy fully in this article, but will refer briefly to the most 
striking diagnostic points in man and beast — intending to 
cover the ground more completely later if possible, when our 
investigations are completed. In man actinomycosis develops 
ordinarily about the maxillary articulation, in the neck, though 
it has occurred in the liver, lungs, stomach and certain glands. 
In external parts the pus generates and localizes in aflat, diffuse 
manner unlike the prominent tumor in cattle. If palped the 
whole mass of pus seems to move in one piece. Slowly the 
skin softens, becomes reddish, perforates, pus exudes, and in it 
may be found the characteristic, yellowish granules. Whilst 
this ulcerative process goes on towards the exterior, it also pene- 
trates deeply attacking all the tissues in the way, such as mus- 
cle, ligament, periosteum, bone, etc. 

Cornil (France) and Babes ( Roumania ) describe six forms 
of actinomycosis in man : 

First. The cervical-maxillary just mentioned. 

Second. The limited neoplasic form, affecting the parotid 
region. 

Third, The thoracic form, affecting the lungs, pleura and 
bronchial tubes, 

Fourth. The lombo-abdominal form, in which the disease 
is in the soft tissues of the abdomen and bones of the lumbar 
region. 



THE BACTERIOLOGICAL WORLD. 33 

Fifth. The pyemic form, in which a number of abscesses 
are disseminated in various organs. 

Sixth. The peritoneal form, in which the lesions are local- 
ized in the peritoneum, sometimes causing perforations of the 
intestines. Ulcers of the intestines were observed. 

Virchow in one autopsy found lesions in the thigh and 
knee. 

Among animals actinomycosis occurs usually only in 
cattle, in which it is known as big jaw, lumpy jaw, etc. Two 
or three cases only are on record as having occurred in the 
horse and in the hog. 




(Tumor of actinomycosis on maxillary of cow.) 
The lesions in cattle vary greatly according to the organs 
and tissues affected. It usually appears at the lower maxillary, 
but is by no means confined there ; it occurs in most of the 
organs. The parasites may cause the production of a fibrous 
tissue, sarcomatous tissue, soft sarcomatous embryonic tissue, 
and sometimes there is even calcification. The specific nature 
of actinomycosis is recognized in those tumors in the form of 
yellowish, grayish and sometimes reddish bodies or tubercles 
ranging in size from a small pinhead to a walnut. The spe- 
cific yellow actinomyces granules or masses are located in these 

bodies. 

The intestinal tissue and the peritoneum frequently pre- 
sent numerous nodules of various sizes ( many about that of a 
small pea ), containing the actinomyces. In the liver such tuber- 
cles are frequent, and even abscesses are occasionally present. In 
the lungs occur often large abscesses and numerous tubercles 
and nodules in which the actinomyces are again found. Some 
affected lungs present the aspect of a case of tuberculosis and 



34 THE BACTERIOLOGICAL WORLD. 

its consequent inflammation, but as distinguishing lesions it 
will be remembered that actinomyces never causes the coagula- 
tion necrosis or caseous degeneration characteristic of tubercu- 
losis. Still, the two affections have been noticed in the same 
individual. 

Another characteristic symptom in actinomycosis is the 
tendency of the abscesses to perforate the skin ; it is so in man 
and beast. 

Judging from the various post mortem examinations which 
I have witnessed, it seems to me that the various forms described 
in man by Oornil and Babes might be applied justly to animals. 
In 42 post mortem examinations in cases of actinomycosis at 
which I was present ( 34 of which were made under the super- 
vision of the Illinois State Board of Live-Stock Commissioners at 
Chicago, August 18, '90), the actinomycosis lesions were found 
distributed as follows : In the lungs in 6 cases ; tongue, 5 cases ; 
liver, 11 cases ; intestines, 19 cases ; post-pharyngial, 7 cases ; 
maxillaries, 35 cases. It will be understood, of course, that 
several had many organs affected. In some, the inferior max- 
illaries, lungs, liver and bowels were diseased, and in others 
only a local external lesion with or without suppuration. 

Processes of infection. — It seems that all the authors 
agree on the mode of infection in man and beast, i. e., by 
lesions communicating with the exterior. These lesions may 
not be accessible to man's eye such as in the intestinal tract, 
the lungs, but they are no less openings to parasites. Ponfick and 
others have mentioned decayed teeth in man as a point 
of entrance of the actinomyces ; he has also mentioned, 
in that connection, a wound of the thumb. . Majocchi 
records a lesion in the skin as having given ingress to the veg- 
etable. Canali and others speak of bronchial wounds in the 
same relation. Cornil and Babes state that it is probable that 
the introduction of the parasites occur by absorption by the 
lymphatics of the mouth ( without any abrasion ). 

In cattle we may safely say that the same means of intro- 
duction cause the infection. We know, in fact, that most of 
cases show lesions about the teeth. At first, perhaps only a 
little nodule may be perceptible at a point on one of the maxil- 
laries, but gradually it enlarges, then it softens, opens outside, 



THE BACTERIOLOGICAL WORLD. 35 

a fistula is formed, and one may sometimes pass a probe through 
to the inside of the mouth — the corresponding tooth or teeth 
having been destroyed more or less. In such cases the parasites 
probably gam entrance between the teeth. 

As in man, then, it is rational to believe that any abrasion 
may give entrance to the vegetable. 

Babes mentions a form of actinomycosis in cattle which 
he terms f ' myelogenous actinomycosis "; it localizes in the 
vertebral column in the interior of the bones in the form of 
purulent tumors with fistula. Perroncito describes another 
kind which he found localized in the skin and subcutaneous 
tissue in the form of sarcomatous and fibrous tumors. Finally, 
Kivolta and Johne saw the actinomyces, in the chronic inflam- 
mation of the spermatic cords of a horse resulting from cas- 
tration. 

The parasite. — Until recently the actinomyces was sup- 
posed to be a fungus ; but Baumgarten, Podvvissowsky, 
Affanassiew, Cornil and Babes, who have studied its biology- 
more deeply and a comparatively short time ago, place it among 
the cladothricea ; hence it would come in the domain of bac- 
terial studies. 




(Unstained microscopic view of actinomyces after Cornil and Babes.) 



36 THE BACTERIOLOGICAL WORLD. 

Artificial cultivation has been generally unsuccessful. Yet 
Johne seems to have succeeded on blood serum, Israel in 
Pasteur's broth, Bostrom on gelatine. 

Inoculation of actinomycosis is possible ; the authors just 
named, and Budjwid (Sankt Petersburger Medecin Wachen- 
schrift, 1888 ), have succeeded several times. Johne has 
transmitted the disease from cattle to cattle by inoculation 
through the skin and obtained local lesions after two months. 
Ponfick succeeded in inoculating it three times through the skin 
and in the peritoneum. Israel inoculated a rabbit successfully 
once through the peritoneum with actinomyces/row a man. 

Complications. — In the abscesses, several authors and the 
writer have found, besides the actinomyces, the streptococcus 
of pus. It is possible, then, that various pus germs complicate 
actinomycosis, and, perhaps, the formation of large abscesses 
depends on the aid of these microbes in destroying the tissues 
gradually altered by the former. 

Sanitary question. — This disease being unquestionably 
parasitic and unquestionably transmissible, it interests the 
public to know what disposition should be made of stock so 
affected. In the first place the diseased stock should be 
destroyed and burned on the farm by the owner as soon as dis- 
covered. This is the safest means of eradicating the disease at 
home so far as known now, for the life history of the parasite 
outside of living organisms is yet a mystery ; it is, therefore, 
impossible to prescribe rational modes of prevention such as 
disinfection, etc., in fields, stables, and the like, particularly 
before the disease makes its appearance. A building contam- 
inated with pus may of course be cleansed in the usual ways. 

It is useless in the majority of cases to attempt treatment 
of the local lesions. True the tumors of the jaw may be excised, 
the wounds cauterized and a local cure may follow. It is even 
possible sometimes to arrest the growth of such a tumor by 
iodine and bi-chloride of mercury injections. But who knows 
that the parasites are not already located in some inaccessible 
organs of the body, such as the liver, the intestines, etc.? 
Should any such chances be taken ? No ; the owner is safer in 
slaughtering at once a case of actinomycosis, and the people 
are thereby saved from grave dangers. 



THE BACTERIOLOGICAL WORLD. 37 

At stock-yards and slaughterhouses the plea is sometimes 
made by owners that a blow or a local injury of some sort is 
the cause of'a jaw tumor that may be visible. This should be 
accepted only with the greatest circumspection ; for such acci- 
dental and sudden swellings of the bone are extremely rare in 
cattle to say the least. 

It has been claimed by interested parties and even by 
physicians and veterinarians that the flesh from many animals 
suffering from actinomycosis is harmless. That may be true 
in part, and again it may not be true in any sense ; it depends 
on the actual distribution of the germs. If the parasite is such 
that it can reach the liver, lodge in the peritoneum, invade the 
muscular fibers of the tongue, cause abscesses in the bones of 
the vertebral column, who will say that the rest of the body 
and the flesh is surely free and safe ? True the muscles with 
the exception of the tongue do not frequently present visible 
lesions, but still they do occasionally, and if the germs migrate 
here and there in the body through the circulation what por- 
tion can be presented to the meat consumer as perfectly safe 
and healthful ? It cannot be safe, and competent rigid inspec- 
tion of stock before killing, and of the carcasses immediately 
after killing are the only safe means of preventing such meat 
from reaching man's stomach. 

The diagnosis of actinomycosis in cattle is easy enough if 
local lesions are present ; there can be no doubt when abscesses 
are formed and pus can be obtained for naked eye and micro- 
scope examination. In man diagnosis may be more difficult, 
but here also it is possible, in many cases, to obtain the specific 
pus showing the actinomyces ; this is the more easy if an 
abscess is external, though it is also possible very frequently to 
make a positive diagnosis of lung cases by examining the 
sputa microscopically. Among the pus may be seen without a 
microscope the fine, more or less spherical, yellow granules 
which contain the parasite and they may also be felt with the 
fingers. If they are masked by the pus mix a little of a potash 
solution with, it to destroy the cells and the actinomyces gran- 
ules may then be seen. If still they are invisible, then a 
microscope must be used. 



38 THE BACTERIOLOGICAL WOELD. 

The technique of the microscopical investigation is too 
long for this article, which, on the other hand, was not 
intended to treat of the histological and microscopical processes 
of investigation. 

In conclusion, I beg to say that the State of Missouri recog- 
nizes the specific, transmissible character of actinomycosis. 
The Veterinary Sanitary Officers have not stopped at the objec- 
tions raised by many that the disease is not contagious. In 
our present knowledge of specific diseases the word contagions 
means only one form of transmission, and, if a disease can repro- 
duce itself by any means in susceptible individuals among our 
food-supplying animals and mankind, it is sufficient to stamp it 
as dangerous to public good and public health. 

Note : — After writing the above, the author came across the 
following, which appeared in the Journal of the Royal Micro- 
scopical Society of England, and which emphasizes the forego- 
ing. This is a criticism on part of a report by Prof. Crookshank 
on Anthrax, Tuberculosis and Actinomycosis : 

"The third disease, actinomycosis, is discussed at consid- 
erable length, but at the same time with perspicuity, and these 
four papers form together the best resume of the subject in the 
English language we have seen. This fungus disease, which 
has been found to attack almost every part of the body, was 
first brought into notice by Bollinger in 1876. The first cases 
were discovered in oxen, but a few years afterwards the same 
disease was found in man. The microscopical appearances 
vary with the anatomical distribution, and also with the rapid- 
ity of the disease. The microscopical appearances of the 
fungus are of two kinds, club-shaped elements which tend to 
arrange themselves in rosettes, and delicate filaments. The 
club-shaped elements, which were the first to be recognized, 
easily stain a red color, while the filaments, which were only 
discovered comparatively recently, stain blue. It is owing 
to this staining difference that Prof. Crookshank was able to 
demonstrate the intimate connection between the two ; for, by 
making careful preparations, he has shown that the filaments 
spring out of the clubs, and that ' the structure of a rosette 
consists centrally of a dense mycelial network, and externally 
of a hymenium of basidia/ 



THE BACTERIOLOGICAL WORLD. 39 

"The question of transmissibility from man to lower ani- 
mals is easily answered in the positive, for calves and rabbits 
have been successfully inoculated with the fungus. 

i ' By the aid of the microscope, the author was able to 
show that i wens ' or f clyers ' are only local manifestations of 
the disease ; previously they were regarded as the result of a 
strumous or tubercular diathesis." 



PRINCIPLES OF AKTI-RABIC VACCINATION. 



BY PAUL GIBIER, M. D. s 

Director of the New York Pasteur Institute. 

To treat of the principle of the vaccination of an infec- 
tious disease whatsoever is, in using a particular example, to 
deal with the question of immunity in general. If, according 
to a theory, which I am not perhaps the first to have emitted, 
the multiple causes of acquired immunity all rest finally on a 
unique principle, we must acknowledge that in certain zymotic 
diseases, such or other of the causes of the acquired immunity 
will have more or less the advantage of the others according 
to the case. 

Concerning rabies, if we examine different experiments 
which have been attempted to explain the mechanism of 
immunity, we see that science is very far from being fixed on 
this point. We will analyze and discuss as briefly as possible 
a few of the researches which have been undertaken on this 
subject. 

Let us glance in the first place to the fact itself. Let us 
take a dog and introduce beneath its skin a certain quantity of 
rabic virus. This simple fact may have four different issues. 
The dog may die after a variable time or it may survive. 
1st, when it succumbs it is of rabies, or, 2nd, it is from the 
intoxication by the rabic substance without being rabid. If it 
survives it may, 3rd, be not vaccinated, or, 4th, have 
acquired immunity by the fact of the injection. We may 
summarize this first exposition in the following table : 



40 THE BACTERIOLOGICAL WORLD, 



( 1st. By hydrophobia. 
' A. Death 1 

( 2nd. By intoxication. 
Inoculation - 

( 3rd. Without immunity 
[ B. Survival •] 

( 4th. With immunity. 

Let us examine now the circumstances in which the cases 
above mentioned may be produced, and let us try and under- 
stand the mechanism thereof. Though the question be, from 
an experimental standpoint, relatively recent, the enthusiasm 
for the experimental method has developed itself so much in 
the last years, that the number of researches which have been 
made on this particular point is already considerable. It is, 
consequently, necessary to have recourse to a classification some- 
what artificial. Let us, therefore, proceed in order. 

1st. After the injection the animal dies of rabies ; it 
presents characteristic symptoms of the disease before dying, 
and after death the virulent principle may be found within in 
its bulb. In this case it is very probable that the needle, if it 
has not extended beyond the limits of the cellular tissue, has 
wounded a nervous filament, and that the virus was put in 
direct contact with the elements of the nervous fiber. The 
experiments of Pasteur, Eoux, Bardach, Helman, etc., and 
mine prove that the virulent element of rabies has a particular 
choice for the nervous tissues. Helman, of St. Petersburg, 
has demonstrated in his experiments, that if the anti-rabic 
injection may be inoffensive when it is made in cellular tissue 
it almost invariably causes death if the needle wounds the 
muscle beneath, or if intentionally the injection is pushed in 
the muscular fibers. It is likely that it is because of the great 
quantity of terminal nervous plaques associated with muscular 
fibers that infection takes place ; it is equally for the same 
reason that the superficial erosions of the skin which expose a 
large number of terminal sensitive nerves are so frequently 
followed by rabies in cases of bites apparently most benign. 
Let us say finally, that it is principally when the quantity of 
virus is light that inoculation has fatal results. When we 
shall come to the study of the case in which immunity is pro- 
duced, we will see, on the contrary, this apparently paradoxal 



THE BACTERIOLOGICAL WORLD. 41 

fact, that the injection of a large quantity of virus may be 
completely harmless. 

2nd. Following the injection the animal dies intoxicated. 
In order that death be produced in this case it is understood 
that a more or less considerable quantity of rabic substance 
must be inoculated. It is not even neeessary that the inocu- 
lated substance be still virulent. Babes and Lepp of Bucarest, 
Roumania, have made experiments in which the injection of 
strong doses of cerebral substance of rabid rabbits, sterilized by 
heat or filtered through porcelain, has determined death after a 
few days. The dogs which received that injection died with 
symptoms of general weakening without symptoms of rabies, and 
the intra-arachnoidian inoculation of their bulb remained with- 
out action on animals on which it was practiced. The filtered 
alcoholic extracts of rabid brains, evaporated in vacuum and 
giving ptomaine reaction, produces in rabbits, guinea pigs 
and mice, after a few days, death without paralysis and witbout 
symptoms of rabies. The extract of brains of animals of the 
same species, but in good health, inoculated in the same condi- 
tions produces no accident. This intoxication is obtained with 
much more difficulty if by small doses gradually augmented 
the organism is accustomed to it. 

3rd. The animal survives and has no acquired immunity. 
I will not insist on this point ; it will be easily understood 
that a single light injection is insufficient to determine immu- 
nity. There is reason to ask how it happens that this injection 
which does not produce immunity does not cause death. We 
will try and explain this fact in the following paragraph, 
where we will study more at length the different theories 
which have been proposed to explain inmunity. 

4th. After inoculation the animal survives ; immunity lias 
been acquired. 

( This point to be continued and discussed at length in 
February number.) 



42 THE BACTERIOLOGICAL WORLD. 

MISCELLANEOUS MATTERS. 



THE DISCOVERY OF KOCH. 



BY PAUL GIBIER, M. D., 

(Director of the New York Pasteur Institute.) 

It is of certain discoveries, as It is of certain ideas ; there is 
a moment when they are in the air. As to the discovery of 
the cnre of tuberculosis, in admitting its existence as an 
accomplished fact, in following, step by step, the progress of 
work executed, and of tbe discoveries made in bacteriology of 
recent date, we see that it was, so to speak, in sight. In 
France Messrs. G-rancher and Martin had arrived, with the aid 
of injections of pure and attenuated cultures of tuberculosis, 
at the production of a certain number of refractory cases in 
rabbits. Messrs. Eichet and Hericourt, by repeated injections 
of sterilized cultures of the bacillus of tuberculosis, have 
rendered rabbits refractory to the inoculation of tuberculosis. 
I have, myself, in 1886 and 1889, begun experiments that I 
had to interrupt at different times ; but in the course of which I 
had made this remark : that the bacillus of tuberculosis 
secreted a substance which is extremely detrimental to its 
development and even to its life. Thus, when we deposit 
bacilli of tuberculosis on a medium where they cannot develop, 
we will, if other germs do not come to destroy them, find 
them aliye again after several months, whilst, on the contrary, 
the more favorable the nutritive material is to the develop- 
ment of the same organism, the shorter will it live therein. 
After a few weeks all traces of life have disappeared ; the 
bacillus can no longer grow on the most favorable culture 
media, and its injection in animals remains negative. 

It may be remarked again that, if we cultivate the bacillus 
of tuberculosis in striae in a tube of gelose a little large, 
colonies do not extend over a few millimeters in width, and 
never reach the edges of the preparation unless an accident has 
dispersed the first colonies developed, over all the surface of 



THE BACTERIOLOGICAL WORLD. 43 

the culture. What prevents the. microbe from invading all 
the nutritive material if it is not a soluble substance secreted 
by the germ and infiltrating the culture medium ? The 
bacillus of Koch, among other species of bacteria studied until 
now, is the one that gives the greatest quantity of soluble 
matter in alcohol and ether. 

After the researches of Hammershley of Bern, the chemical 
analysis of the bacilli give water 88.82 per cent., solid sub- 
stances, 11.18 per cent. Of these solid substances 22.7 per 
cent, are soluble in alcohol and ether. Inoculations of these 
extracts practiced on animals have shown that they contained a 
toxic matter with tetanic properties. It is to be noted that in 
persons inoculated with Koch's lymph, besides the febrile 
movement, the patients exhibit occasionally cramps and stiffness 
in the limbs. 

These observations, and a few experiments that I have 
made, as well as the facts presented by Messrs. Grancher and 
Martin, Hericonrt and Eichet, authorize us to give this opin- 
ion : That the albumenoid extracts of the cultures of the bacillus 
of tuberculosis form the base of the lymph of Dr. Koch, and if 
the latter does not hasten to publish the complete results of his 
investigations the priority of the discovery might be contested. 



STUDY OF TUBERCULOSIS IN FRANCE. 

The great excitement, due to the announcement of Koch's 
discovery of an agent capable of awing tuberculosis, has, it 
seems, completely overshadowed a public statement, made later, 
of the admirable results obtained by scientists in Pasteur's 
Institute, in their experiments made with the object of pre- 
venting the same disease by vaccination. 

Messrs. J. Grancher and H. Martin, soon after the report of 
Dr. Koch, as made at the Medical Congress of Berlin, published 
a letter explaining that on the 19th of November, 1889, they 
deposited at the office of the Academy of Medicine ( Paris ) a 
sealed document, giving their success in arresting, for a long 
time, the evolution of tuberculosis in rabbits. The process of 
vaccination was after the anti-rabic system of vaccination of 



44 THE BACTERIOLOGICAL WORLD. 

the great French master, Pasteur, that is, by the inoculation 
of graded virus. Virus number 1, say, may represent the most 
virulent, and 2, 3, 4, 5, 6, 7, 8, 9, 10, being successively 
decreasing. The introduction of the material was done as in 
hydrophobia, by beginning with the weakest, and at intervals 
introducing some other stronger, and again still stronger. 
After such vaccination, even the introduction of the strongest 
tuberculous virus, according to the quickest method of infec- 
tion known, namely, introduction of tuberculosis bacilli in the 
veins failed to produce the usual specific lung, liver and spleen 
lesions. 

Thus we have, it seems, another method in sight by which 
mankind may combat the terrible ravages of tuberculosis ; and 
it is an important one, too, which promises great good to the 
people. 

"We hope that Messrs. Grancher and Martin will be suc- 
cessful in their endeavors to give the world a prophylaxis 
against the worst plague to which man and beast are liable. 
Thus, with a cure from the German masters, a preventive from 
the French savants, the universe will be doubly blessed, and the 
nations owe a still greater debt of gratitude to the discoverers 
and science. — Ed.] 



THE TREATMENT OF SKIN DISEASES IN GENERAL PRAC- 
TICE. 



BY J. W. BANKS, M. D. , 

Columbia, Mo. 

There is scarcely anything more troublesome to the general 
practitioner of medicine than the great variety of skin affections 
that come to his notice. Firstly, the patients in such cases 
use usually all sorts of nostrums before presenting themselves 
for treatment by a competent physician ; secondly, when they 
do request treatment they scarcely ever will follow a regular 
course of treatment ; and, lastly, the general practitioner is not, 
as a rule, so situated as to study the cases as closely as they 
should be or to follow the results of his treatment as a spe- 
cialist can. As a result he frequently finds himself confronted 



THE BACTERIOLOGICAL WORLD. 45 

with difficult and tedious maladies, and he is forced, too often, 
by the circumstances and facts alluded to, to experiment more 
or less before success is attained. At least, I can conscien- 
tiously say, that such has been my experience in many years of 
practice. 

Since the progress made in medicine, because of the dis- 
covery of the action of germs on the economy, it strikes me 
that many skin affections, besides those known to be primarily 
due to microbes, are at least much complicated by the access 
of injurious germs into the seat of the diseased localities of 
the tegument. This I believe to be surely the case in the 
eruptive maladies, whether primarily due to a specific virus or 
not. Therefore, it seems to me that these possible factors 
ought to be taken into account in medical or surgical treat- 
ments, and such medicines used as combine the soothing, heal- 
ing and sometimes the stimulating effects with the antiseptic 
and bactericide properties. In the last year I have applied 
this rule in my practice uniformly, and I am pleasantly con- 
fronted with a list of good results, fifty per cent, better than 
ever before ; successes indeed, where heretofore I failed utterly. 
Of course I have not abandoned constitutional treatment where 
such seems absolutely necessary, but my local applications have 
been much modified, and many cases in which two years ago I 
thought internal treatment indispensable have since yielded with 
only local medication. The principle I follow is first perfect 
cleanliness and then applications preventing the growth of 
germs in the diseased parts, instead of using the ordinary skin 
ointments so-called, under which microbes often flourish. But 
these cases explain themselves. 

I herewith present a few instances of inveterate, per- 
sistent affections, some of which I had previously failed to 
cure with the time-honored local prescriptions in conjunction 
with the iodides, mercurials, and the arsenical preparations. 

I had long since recognized the beneficial action of vege- 
table tar in skin diseases, but never before had I encountered 
a preparation containing it, which was not objectionable and 
often valueless because of its color, its sticky and granular 
condition, and pungent odor. The preparation I have used so 
largely of late is Tarro-Petrolene ( or Petrolene Comp. "No. 1, 



46 THE BACTERIOLOGICAL WORLD. 

refined), in which tar and petroleum are so beautifully and 
homogeneously blended as to form a veritably desirable salve, 
unequaled in my estimation either as an ointment or a base of 
ointment for maladies of the skin and scalp. 

Patient No. 1. — Curtis E., scald head, Orustea Lactea, 3 
months' standing ; attack very obstinate with complete loss of 
the hair. Yielded very readily to an application of Petrolene 
Oornp., or Tarro-Petrolene No. 1, each night for one week. 

Patient No. 2. — Mattie B., congenital syphilis. Applied 
Petrolene Comp. over the eruption which was all over the 
body with great loss of epidermis about nates and groins. 
Kept up application for 2 weeks with minute doses of calomel 
internally. Syphilitic eruption all disappeared and child now 
apparently in good health, assimilates food readily and growing 
very fleshy. 

Patient No. 3. — Cornelia E., aged 40 years, with local 
tetter on the forearm ; a washerwoman. Eruption very irritated 
by contact of soap and hot water. G-eneral health good. Used 
Petrolene Comp. for 2 weeks every night before going to bed, 
and all appearance of tetter has disappeared. 

Patient No. 4. — John D., secondary syphilis. Eruption 
scaly, extended over face and body. Used Petrolene Comp. 
over the eruption with Iodide Pot. and Per-Iodide of Mercury 
internally, and in 4 weeks no syphilitic symptoms. 

No. 7. — Eczema of the face in child 4 years old. Yielded 
to application of Petrolene Comp. applied 2 weeks. 

No. 8. — George E., Acne of face. A very obstinate case. 
Been treating him for 3 weeks with Petrolene Comp., with 
Iodide Pot. and Per-Iodide of Mercury ; patient almost entirely 
cured. 

The above are a few of the cases I have kept notes on in 
treatment, and where treatment has been fully adopted 
satisfaction has been complete or encouraging. 



THE BACTEKIOLOGICAL WORLD. 47 

SCIENTIFIC GLEANINGS. 

Structure oe bacteria and allied organisms.* — 
Prof. 0. Butschli deals with the finer structure of bacteria in 
a monograph, the object of which is to demonstrate the propo- 
sition that these organisms are nucleated bodies. The ulti- 
mate structure of these minute bodies is divided into two parts, 
an outer harder layer and an inner softer part, the central 
body. The former is further distinguished from the central 
body by being less stainable with the ordinary pigments. In 
both a reticulated appearance is discoverable with high magni- 
fying powers, and on section this reticulation or network 
imparts a honeycombed appearance to the object. At the 
points of intersection of the network are frequently seen 
red globules of variable size. These red globules, which are 
very frequent in the sulphur bacteria, are supposed to consist 
of sulphur in some vascid condition. Their exact significance 
seems doubtful. One point on which the author expresses him- 
self confidently is that when a flagellum is present it is contin- 
uous with the outer layer. The central or chromatic part of 
the micro-organism is to be regarded as the nucleus. 

Micrococcus versatilis.* — This micro-organism, which 
Dr. 0. Delgado and Dr. 0. Finlay presume to have some direct 
connection with the appearance of yellow fever, has been 
obtained by the usual methods, not only by these authors, but 
also by Dr. Sternberg and others from the juices and tissues of 
persons affected with or dead of yellow fever. The authors 
have obtained it from the serum of artificial blisters, and 
Sternberg from the skin of healthy, but unwashed, persons liv- 
ing in places where the fever is endemic. The name originally 
given to this micrococcus by Delgado and Finlay was " Tetra- 
genus febris flavse," but they now accept the new name pro- 
posed by Sternberg. 

The best procedure for obtaining micrococcus versatilis is 
to inoculate peptonized gelatin and keep it in the incubator at 
a temperature of 30°-32°. When a deposit forms at the bottom 



* Journal of Royal Microscopical Society. 



48 THE BACTERIOLOGICAL WORLD. 

of the gelatin, gelose in Esmarch's tubes is inoculated there- 
with, and the tubes kept at a temperature of 30°-32°. Colo- 
nies began to appear in from two to six days, according to the 
season of the year and the activity of the germs cultivated. 
The colonies are round, with smooth edges, transparent, of 
straw-yellow color, but become opaque as they grow older. 
The colonies below the surface are more fusiform and deeper 
colored. 

The name versatilis was suggested by the different appear- 
ances presented by the colonies, their variable color, and the 
diverse sizes of the micrococcus, which is chromogenous, and 
develops in true tetrads. 

Action of the gastric juice on pathogenic mi- 
crobes.* — MM. J. Strauss and R. Wurtz have examined the 
action of the gastric juice of the dog, man and sheep on the 
bacilli of tubercle, anthrax, enteric fever and cholera. The 
gastric secretion was placed in test-tubes, inoculated with the 
microbes, and kept at a temperature of 38° C. After various 
periods of time, inoculation experiments in guinea-pigs and 
rabbits showed that tubercle bacilli survived the action of the 
gastric juice up to six hours sufficiently well to produce a gen- 
eral tuberculosis. From 8 to 12 hours a tubercular abscess was 
formed at the inoculation site, and this quickly healed. After 
18 hours the bacilli were either dead or had lost their viru- 
lence. Anthrax bacilli without spores were killed in 15 to 20 
minutes, and the spores died after thirty minutes. Under 
similar conditions typhoid bacilli died in 2 to 3 hours, and 
cholera bacilli after 2 hours. 

Further experiments with hydrochloric acid in the propor- 
tion of • 9, 1*7, and 3 per thousand on anthrax, cholera and 
typhoid bacilli gave similar results to those obtained with gas- 
tric juice. Hence the authors conclude that the antiseptic 
effect of the gastric juice is to be ascribed to its containing 
hydrochloric acid. 



* Journal of Royal Microscopical Society. 



THE BACTERIOLOGICAL WORLD. 49 

REMEDIES AND CLINICAL REPORTS. 

Methyl violet. — This substance, prepared now by 
Mercks, chemically pure, under the name of {( Pyoktanin," 
seems an excellent antiseptic in all cases of suppuration. 
Gonorrhea, ulceration of the cervix and uterus, shankers, 
granulated eyelids, etc., are reported as having been success- 
fully treated, by several competent physicians. The substance 
is used in the average proportion of 1 to 2,000. It is not 
irritant, scarcely toxic, and yet strongly bactericide. The first 
experiments with it were made in Strasburg. 

Tarrq-petrolene (Syn.: Petrolene Compound). — This 
ointment (used also as a base) seems a remarkable, healing, and 
at the same time antiseptic preparation in skin diseases. 
Reputable physicians report cures of tetter, eczema and scalp 
and skin affections of a parasitic nature, or with bacterial 
complications. This ointment is perhaps the only one contain- 
ing vegetable tar so smoothly blended or incorporated. The 
purified Petrolene Compound or Tarro-Petrolene, grade No. 1, is 
amber color, and has rather a pleasing odor ; the crude, or No. 2, 
is darker, and used in veterinary practice. In animals it is 
reported as efficacious in skin diseases also. It is frequently 
used alone or as a base for iodine ointments in so-called 
scratches in horses. 

Katharmon. — This antiseptic or bactericide lotion is 
getting in general use in various pathological discharges of the 
mucous membranes. Its success seems to be greater in nasal 
catarrh and vaginal catarrh. Altogether, very favorable reports 
come in from practitioners. 

Iodoform. — This drug, once thought to be highly anti- 
septic, is losing ground and is being replaced mostly by Iodol, 
which has much the same properties, is apparently a superior 
bactericide and has none of the very objectionable and disagree- 
able odor of the former. 

Pambotako. — In Italy this substance is rapidly taking the 
place of quinine in malarial fever, and well it may, for, judging 
from most trustworthy reports, it has very remarkable effects. 



50 THE BACTERIOLOGICAL WORLD. 

In many cases — even some of inveterate character — only one 
single dose was sufficient to cure the disease radically. In fact, 
in its use by the Italians and French who employed it, it seems 
the rule to give but one or two doses even to patients who had 
suffered for months, periodically or constantly. Pambotano is 
not a substitute for quinine in case of, periodicity, but it seems 
very nearly a specific in malaria. Why not try it in this 
country? 




CATARRH. 



Dr. J. J. Stephens, of Clinton, Mo., recommends the 
following in nasal catarrh. 

First he prescribes a mercurial purge, following it with a 
saline cathartic. Then he uses douches, in which Katharmon 
seems the principal factor. After the action of the cathartic 
he uses the following : 

1^ Katharmon, 5 ss. 
Sodii Chloride, grs. ii. 
Aq. Distil, g iss. 
Sig. — One tablespoonful in 12 oz. of warm water as a 
douche. 



ANTISEPTIC AND DEODORIZING MOUTH WASH 

F, W. Peck, druggist, Columbia, Mo., highly recommends 
the following lotion for the mouth : 

ty Acid. Carb., m xx. 

Acid. Acetic, 5 i- 

Tinct. Myrrh, 5 ii- 
Honey. 

Listerine, aa. § i. 
Aqua dist. ad. qs., § viii. 
Misce. 



THE BACTERIOLOGICAL WORLD. 51 

SKIN ERUPTIONS AND COMPLICATED, INDOLENT, REBEL- 
LIOUS SKIN ULCERATIONS. 

Dr. P. Paquin prescribes the following in such cases and 
recommends it in man and beast : 

1^ Pyoktanin, grs. ii. 

Resorcine, 5 i- 

Tarro- Petrolene, 

(Petrolene Co. No. 1 or No. 2) § jy. 
M. Ft. ring. 
Sig. — Wash thoroughly, wipe dry and then apply ointment 
in friction daily. 



TECHNIQ UE. 

Preparation of nutritive agar.* — In the course of 
experimental work Dr. V. A. Moore found that when the 
stems of agar were cut into small pieces and boiled in a fluid 
containing no coagulate material, that it was entirely broken 
up and the soluble portion dissolved. The insoluble particles 
that remained suspended in the liquid were easily and com- 
pletely removed by the addition of egg albumen, and subsequent 
boiling and filtering. From these facts a method for the prep- 
aration of nutritive agar was derived, which consists in first 
preparing the neutralized beef-infusion-peptone, and thus get- 
ting rid of all coagulable material before the agar is added. 
This process is effective in greatly diminishing the time and 
attention required for the preparation of this medium. The 
medium can always be made of the same consistency, as all of 
the agar that is added is dissolved. It remains free from pre- 
cipitates when sterilized, and its nutritive qualities are as 
favorable to bacterial growth as when it is prepared after the 
original method. 

(1) The preparation of the beef-infusion-peptone. — The 
method of preparing this liquid is practically the same as that 
already in use in most laboratories. Finely chopped or ground 
beef (freed from fat) is macerated in distilled water for from 
12 to 18 hours in a cool place. The distilled water is added in 

*From the Journal of the Royal Microscopical Society. 



52 THE BACTERIOLOGICAL WORLD. 

the proportion of 200 com. to each 100 grams of beef. On the 
following day the liquid is separated from the meat by strain- 
ing it through a coarse linen. The simple beef-infusion thus 
obtained should be equal in quantity to the amount of water 
added ; if it is not, the deficiency can be restored by the addi- 
tion of distilled water. To the beef-infusion is added one per 
cent, peptone, 1-2 per cent, sodium chloride ; and if it is desir- 
able to make it alkaline, a sufficient quantity of a normal solu- 
tiou of sodium carbonate to give it a weak alkaline reaction. 
The liquid is then boiled for thirty minutes in a water-bath, 
cooled, filtered and distributed in Erlenmeyer flasks plugged 
with cotton-wool. If only a small quantity of agar is to be 
made at once, 250 ccm. is found to be a very convenient quan- 
tity to put in each flask. It is then sterilized by boiling for 
one hour each day for three consecutive days. It need not be 
sterilized if it is desirable to prepare the agar at once. As the 
beef-infusion-peptone is also employed as a liquid medium in 
the cultivation of bacteria, very little time is lost in preparing 
an extra quantity of this liquid to be used in making the agar. 
( 2 ) The preparation of the agar, — To an Erlenmeyer 
flask ( a glass beaker or agate or iron vessel may be used ) con- 
taining beef-infusion-peptone, as prepared above, 1 per cent, of 
very finely chopped agar is added. The flask is then placed in 
a water-bath and boiled vigorously for two hours. At the end 
of that time the agar is dissolved, and the liquid is allowed to 
cool. When a temperature of 40°-45° C. is reached, the white 
of egg is added in the proportion of one egg to 250 ccm. of the 
liquid. After the albumen is thoroughly mixed with the 
liquid agar it is returned to the water- bath, and again boiled 
for two hours. It is of much importance that the albu- 
men is evenly distributed throughout the mass before it is 
coagulated. It is now ready to be filtered. The egg albumen 
is coagulated in very firm masses, leaving the liquid, perfectly 
clear. The coagulum is removed by filtering the liquid 
through fine Japanese filter-paper or a layer of absorbent cot- 
ton, as a 1 per cent, solution of the agar does not pass readily 
through ordinary filter-paper. Should a weaker solution of 
the agar ( 1-2 to 3-4 per cent.) be desired, its filtration can be 



THE BACTERIOLOGICAL WORLD. 53 

accomplished by the ordinary method. A hot filtering appara- 
tus is not necessary. The clear filtration is now ready for dis- 
tribution in sterile cotton-plugged tubes. 

The agar is sterilized by discontinuous boiling in a closed 
water-bath for three consecutive days. If small tubes have been 
used containing not more than 7 ccm. each, five minutes' boil- 
ing each day is sufficient. If larger tubes are used, they should 
be boiled for a longer time. Or it may be sterilized by steaming 
each day for from five to ten minutes after the agar has become 
liquefied for the same number of days. After its sterility has 
been tested by allowing it to stand in an incubator for several days, 
it is ready to be stored away until required for use. It has been 
customary in this laboratory, in order to prevent the evaporation 
of the agar by long standing, to dip the lower end of the 
cotton-plugs in hot sterilized paraffine, and to store the tubes in 
a cool, moist chamber. 

Methods of recognizing cysticerci of taenia sagi- 
nata.* — M. A. Laboalbene has a note on the means of recogniz- 
ing the cysticerci of taenia saginata, which are the cause of 
"measles" in veal and beef, and which are often so difficult to 
detect on account of the rapidity with which they dry on 
exposure to the air. He finds that meat which has become 
quite leathery will easily reveal the cysticerci, if it contain any, 
by being placed in water acidified with acetic or nitric acids, 
or in a mixture of water, glycerine and acetic acid. By this 
means the parasites can always be detected, and if the meat 
be carefully heated to 50° or 60° C. it is always fit for human 
food. 



LABORATORY NOTES. 

New York, October 15, 1890. 

Dr. Paul Gibier, Director of the New York Pasteur Insti- 
tute, begs to inform you of the results of the preventive inocu- 
lations against hydrophobia performed at this Institute since 
its opening (February 18, 1890). 

To date 610 persons, having been bitten by dogs or cats, 
came to be treated. These patients may be divided into two 
categories : 

^Journal of the Royal Microscopical Society. 



54 THE BACTERIOLOGICAL WORLD, 

1st. For 480 of these persons it was demonstrated that 
the animals which attacked them were not mad. Consequently 
the patients were sent back after having had their wounds 
attended, during the proper length of time, when it was neces- 
sary. Four hundred patients of this series were consulted or 
treated gratis. 

2d. In 130 cases the anti-hydrophobic treatment was 
applied, hydrophobia having been demonstrated by veterinary 
examination of the animals which inflicted bites, or by the 
inoculation in the laboratory, and in many cases by the death 
of some other persons or animals bitten by the same dogs. 
All these person's are, to-day, enjoying- good health. 
In eighty cases the patients received the treatment free of 
charge. 

The persons treated were : 

64 from New York, 3 from Pennsylvania, 1 from Ohio, 

12 from New Jersey, 2 from N. Hampshire, 1 from Arizona, 

12 from Massachusetts, 2 from Georgia, 1 from Iowa, 

8 from Connecticut, 2 from Texas, 1 from Nebraska, 

9 from Illinois, 1 from Maryland, ,1 from Arkansas, 
3 from Missouri, 1 from Maine, 1 from Louisiana, 

3 from N 'th Carolina, 1 from Kentucky, 1 from Ontario, (Can.). 

With kindest regards of the Pasteur Institute, 

Paul Gibier. 



NE W P UBLIGA TI0N8. 

The text-book on Comparative Physiology, by Professor 
Wesley Mills, of McG-ill University, Montreal, Canada, is 
timely. In the last two years two excellent and much-needed 
works on the subject have been written : one by Prof. Mills 
and the other by Prof. Smith, of Pennsylvania University. 
Both filled a gap in English medical literature which had been 
felt for many years by the students, and practitioners of com- 
parative medicine and surgery, but the smaller volume of Prof. 
Mills, recently published by D. Appleton & Co., is more 
compact and perhaps the most suitable text-book ever printed 
on the subject of which it treats. P. P. 



THE BACTERIOLOGICAL WORLD. 55 

AD VERTISEMENT. 

Our magazine we shall endeavor to keep of the highest 
character, scientifically and practically, and we must refuse, 
with due respect, all advertisements of medical preparations, 
the legitimacy of which is questionable, or which do not 
receive the indorsement of some reputable member of the 
medical profession. Not having published this fact in our 
prospectus, we beg to apologize for having returned the MSS. 
of a gentleman whose "ad" we had solicited before knowing 
the nature of the preparation to be advertised. On the same 
principle we must refuse space to questionable medical institu- 
tions of any kind. 

But, on the other hand, believing that the duty of a 
physician is to relieve sufferings, and to prevent them and cure 
them by any or all legitimate methods, we consider it our 
duty not only to publish advertisements, but also t to aid with 
all our might in establishing the actual virtues and properties 
of worthy articles among the medical world and the public, 
by the publication of clinical reports from authorized sources. 

Our readers, therefore, may rely on whatever may appear 
of such character in any part of this magazine. 

We issue 5,000 copies of this edition and shall issue at 
least 1,000 copies monthly hereafter, reaching every state and 
territory in the Union. 

We respectfully solicit a share of the patronage of all 
deserving institutions, firms and individuals who usually adver- 
tise in medical and scientific journals. 

The Bacteriological World. 



56 THE BACTERIOLOGICAL WORLD. 

IMPORTANT NOTICE TO SUBSCRIBERS, 
EXCHANGES, ETC. 

We had the intention of mailing nearly all of our first 
5,000 copies to a selected list before January 1st, but, since 
circulating our prospectus, the medical public has so generously 
and so promptly responded, that we have decided to distribute 
about 3,000 within that date, including the subscribers then 
listed, and to mail the remainder gradually, less 300 copies, as 
subscription may be received, within the months of January 
and February, thereby affording opportunity to examine the mag- 
azine, and keeping faith with our engagements. Exchanges 
will kindly notice. If, after the whole issue is exhausted, we 
still receive subscriptions we may decide to issue another 
edition. 

We are now making arrangements with the publishers of 
some of the leading medical and scientific periodicals to the 
end of offering the Bacteriological World with one or 
more magazines at reduced terms. Our publication being a 
necessary adjunct to medical, hygienic, veterinary and certain 
other scientific journals, we hope by such means to 'bring our 
infant more prominently before the interested public. 

Send postal notes, money orders or checks on New York 
or St. Louis. Never send check on local banks as they cannot 
be collected without paying 25 cts. exchange. 

Editor and Publisher. 




JVI F? . GEORGE JVHLLEF^ STERNBERG. 




JU 



CA 




A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies. 



Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological World, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to by Dr. 
T. J. Turner, Mexico, Missouri, to whom remittances should be made. However, 
all moneys sent to the Bacteriological World Co., Columbia, Mo., will be received 
and promptly acknowledged. 

Authors of original articles published in this magazine will be entitled to twenty 
copies of the number containing it, provided, that they request the same in writing 
when sendiDg the communication. Reprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. 

Discontinuation :— At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first of next January. 

COLLABORATORS : 

Paul Gibier, M. D., New York. Prof. L. Bremer. M. D., St. Louis, Mo. 

Prof. W. T. Belfield, M. D., Chicago. J. W. Stickler, M. D., Orange, N. J. 
Prof. A. W. McAlester, A. M., M. D., Prof. Paul Schweitzer, Ph. D., 

Missouri University Med. Dep't. Missouri University. 

Prof. T. J. Burrill, Ph. D., Illinois University, 
Director of Department of Vegetable Mycology. 



U. S. A, FEBRUARY, 



1,2. 



EDITORIAL. 



KOCH'S REMEDY AND OTHER TREATMENTS OF 
TUBERCULOSIS. 

The press has been so full of this subject of late that it 
seems superfluous to add anything more. Yet this question is 
directly in line with the duties of the Bacteriological 
World, and it devolves upon us to review it to date. 

The excitement has now subsided, the confusion has disap- 
peared in a great measure, and we can see the facts as they are. 

In a nutshell the situation is this : 

First. Koch's lymph is successful in the treatment of 
localized tuberculosis, particularly in the comparatively mild 
form known as lupus. 

Second. It may be beneficial in a measure in cases where 
the baccilli have attacked the lungs mildly. 



58 THE BACTERIOLOGICAL WORLD. 

Third. It may be dangerous in serious cases> when 
internal organs are involved. 

Fourth. It is dangerous in the hands of indifferent, care- 
less practitioners. 

Fifth. It is dangerous to use it indiscriminately for diag- 
nostic purposes. 

There is no question, however, that the Koch discovery is 
one of great value from a scientific standpoint ; but much is 
to be done in a practical sense to make it as useful as we had 
hoped it would be, at an earlier date, when we were informed 
by the Germans, that the state was preparing to supply the 
drug trade with Koch's lymph under the name of " parato- 
loid." We had expected that the scientists interested would 
not allow this to be done unless they were sure of safe results. 
It seems that this commercial aspect of the case was prema- 
turely countenanced. 

On the whole, from a v rational basis, one could scarcely 
expect, at this early date, any more favorable result than was 
obtained. All things considered, Koch's remedy stands on a 
basis which insures its earnest, respectful consideration and 
trial by the most trustworthy of the medical fraternity, with a 
good prospect of success in certain cases of tuberculosis. Only 
when the full properties and limitations of the new agent have 
been clearly established will it be possible to realize the benefit 
of it to mankind. 

But, whilst we, American physicians, make so much of 
this achievement, we have neglected to give the deserved 
encouragement to home investigators who have made striking 
discoveries in the very same line of work. 

Koch's great name carried his results to every part of the 
world, and they were received with confidence, because it came 
from Koch ; it would have been different had it been from a 
more humble origin. Still, with all his reputation, Koch's 
achievement should not so overshadow every other man's 
works as to relegate them to the rear. It is particularly 
humiliating, and positively painful, that deserving American 
scientists who, themselves, are quoted abroad as authorities, 
barely receive recognition from the American press in general. 



THE BACTERIOLOGICAL WORLD. 59 

Dr. Samuel G. Dixon modestly announced, in 1889, that 
lie had succeeded, by a scientific process, in producing immu- 
nity in small animals against tuberculosis, by the inoculation 
of a solution of certain principles extracted from the baccilli 
of tuberculosis in a given morphological condition. The 
immediate effects of this inoculation were very similar to those 
produced by Koch's lymph. 

Dr. Kraft, of Cleveland, Ohio, is quoted as having pro- 
duced practically the same thing with a substance that he 
terms lymph. 

Dr. D. E. Salmon, Bureau of Animal Industry, and his 
colleague, Dr. Smith, announced, in 1887, their experiments 
in producing immunity against a bacterial disease, hog- cholera, 
by the inoculation of sterilized cultures ; and, in 1889, Dr. 
Schweinitz, of the same institution, reports the production of 
immunity with chemical substances formed during the growth 
of baccillus of hog-cholera. We reprint the latter's report in 
this number. 

Dr. Frank S. Billings has applied for two years a process 
of inoculation to prevent swine-plague, which he claims is also 
a chemical substance, though, unfortunately, no literature has 
ever been published by the Doctor verifying this point for the 
benefit of science. 

Lastly, Drs. Shurley and Gibbes, of Detroit and Ann 
Arbor, Michigan, have formulated prescriptions of a purely 
chemical nature, which have had, it seems, success even more 
remarkable than the Koch remedy. 

Ail these are scientific results brought about by Amer- 
ican brains, and it is rather strange that we do not encourage 
more our own deserving enterprising men ( who, by the way, 
are not all backed by government subvention, and, therefore, 
often work under great difficulties). 

Treat everyone with equal justice and consideration ; give 
credit fully " to whom credit is due," and let us remember that it 
is not necessary to live abroad to possess qualifications 
necessary to succeed in medical and scientific investigations. 
" Science has no home, but the universe at large." 

The enthusiasm that arose in the breast of every lover of 
medicine and human nature spread wildly throughout the 



60 THE BACTERIOLOGICAL WORLD. 

universe, when it was learned that no less a man than Koch, 
himself, had announced his belief in being able, in the near 
future, to cope with tuberculosis ; few were capable of resisting 
the temptation of expressing their gratitude to a man who 
prophesied such a boon to humanity. 

The Bacteriological World expressed its feelings in the 
same vein at the prospects. Now, since learning more of the 
foreign discovery, and more of the results obtained by our own 
countrymen, we do not hesitate to say that our home achieve- 
ments are equal to the Germans', to say the least ; that some of 
ours seem to have priority in the line of Koch's discoveries, 
and that we feel equally enthused and grateful when we realize 
fully the depth and potentialities of the American discoveries. 
In weighing everything, we find, on calm reflection, that all 
these results have advanced medicine at a giant pace towards 
the solution of the question of tuberculosis. 



COMPARISON OF KOCH'S REMEDY AND SHURLEY AND 

GIBBES' TREATMENT. 

The Koch remedy, as explained by himself, though it is 
not absolutely clear just what principles it consists of, and the 
explanation does not reveal the process of the preparation 
sufficiently to allow exact repetition by others, yet it is given 
that the basis of it is an agent produced by the bacillus of 
tuberculosis itself. The action upon which its value rests is 
the disintegration and the exfoliation of the morbid tissues. 
The affected parts are destroyed. Now, this fact at once limits 
the utility of Koch's remedy to a comparatively few cases. 
For, in fact, how can we conceive that the disintegration, 
death, of a portion of an important internal organ may occur at a 
given moment with impunity. Such destroyed portions must 
act as foreign bodies, and must either be circumscribed by new T 
tissues by nature's efforts, or absorbed or extirpated. In either 
case it is a serious question, and in many, the first sequence, if 
it did occur, would constitute a grave interference ; the second 
does not seem possible in the majority of cases ; whilst the last 
could scarceJy be practiced. Would it be usually practical, for 



THE BACTERIOLOGICAL WORLD. 61 

instance, to remove the exfoliated parts from a liver, or spleen, 
or intestines, or peritoneum, or the heart, or the brain, or the 
lungs, more or less affected or filled throughout with tubercles ? 

Dr. Heneage Gibbes, in a recent paper before the Medical 
and Library Association of Detroit, Michigan, draws attention 
to these points in a forcible manner. 

Furthermore, he presents unanswerable arguments against 
the application of the remedy in many cases of tuberculosis of 
the lungs, by his very accurate division of the forms of the 
disease. Basing a classification on morbid histology, we must 
admit, as Prof. Gibbes so clearly points out, that there are two 
distinct forms of consumption ; one in which the baccilli are 
numerous and scattered through a variable portion of tissue ; 
the other, in which the disease consists of nodules, made of new 
formation in which the baccilli are more scarce. This fact 
precludes the use of Koch's remedy in the first kind of con- 
sumption, and limits it in the other. Suppose the half of one 
lung is affected with the acute, inflammatory, scattered, 
miliary tuberculosis, what would be the result of a treatment 
causing the death of this part in total ? 

And now comes Yirchow, who says that Koch's lymph 
may, under some conditions, cause, in a degree, the generaliza- 
tion of tuberculosis where it before was localized. 

Again appear authoritative statements to the effect that 
for diagnostic purposes, the reaction of Koch's lymph may 
prove dangerous. What then is there left, in which Koch's 
remedy, so far as its action has been explained, may be truly 
beneficial ? Local tuberculosis, viz.: Lupus, cutaneous, bone, 
gland, articular* and incipient tuberculosis. 

The fact remains, however, that the remedy does actually 
cure tubercles, and this, in time, may lead to more practical 
results in the treatment of consumption, generally speaking. 

Shurley and Gibbes' treatment presents less objections ; 
it would seem that it could reach more cases, the only 
condition being that the destruction be not too great in the 
affected parts prior to treatment. In this case it is the cause 
alone that is removed and prevented from acting. The germ is, 
by the chloride of gold injection, so affected, that it ceases to 



62 THE BACTERIOLOGICAL WORLD. 

multiply and scatter, and the tissues may regain their equilib- 
rium. There is no dead matter suddenly left to act as foreign 
body ; there is no dangerous reaction. The results have been, 
from all accounts, very gratifying and promise more extensive 
application, perhaps, than any other remedy advanced to this 
date. 

But, even if Koch's remedy should prove, in time, to be 
the least beneficial of all that have or may be brought to light, 
we must not forget that much of the honor and credit of the 
success reflects on the celebrated German physician, because all 
are and must be based on his discovery of the germ of tubercu- 
losis and its properties. 



STERNBERG. 



George Miller Sternberg, Major and Surgeon, U. S. A. 
Born in New York state, June 8, 1838 ; graduated in medicine 
at the College of Physicians and Surgeons, ISTew York City, 
in the spring of 1860 ; appointed Assistant Surgeon, U. S. A., 
May 28, 1861 ; Major and Surgeon, U. S. A., December 1, 1875 ; 
saw active field service during the War of the Rebellion and has 
seen much service on the frontier since ; appointed a member of 
the Havana Yellow Fever Commission of 1879 ; delegate from 
the United States to the International Sanitary Conference of 
Rome, in 1885 ; detailed by the President to make investigations 
relating to the cause and prevention of yellow fever in 1887 ; in 
pursuing this investigation visited Rio de Janeiro and Vera Cruz, 
and spent the greater portion of two summers in Havana ; elected 
President of the American Public Health Association in 1886. 
Is an Honorary Member of the Epidemiological Society of 
London ; of the Royal Academy of Medicine of Rome ; a*nd of 
the Academy of Medicine of Rio de Janeiro, Is a Fellow of 
the American Association for the Advancement of Science ; 
and of the Royal Microscopical Society of London ; Associate 
Member of the Societe Francaise d' Hygiene, etc., etc.. 

Dr. Sternberg is one of the oldest and most distinguished 
bacteriologists in America. His reputation in this country and 
abroad is deservedly high. He has contributed, we believe, 



THE BACTERIOLOGICAL WORLD. 63 

more than any other one man of oar country to bacteriological 
literature. His past services in the medical world, and to the 
country at large, are great indeed, and it is to be hoped that we 
may continue to reap the benefits of his knowledge aud 
experience for many years to come. 

Sternberg is the author of numerous, excellent publica- 
tions bearing on bacteria, among which we note as most 
important: "Bacteria" ( Magnin & Sternberg); "Photo- 
Micrographs, And How to Make Them"; "Malaria and 
Malarial Diseases" ; " Disinfection and Disinfectants." 



NEW COLLABORATORS. 

We take much pleasure in announcing to our readers the 
names of Dr. H. P. Loomis, Professor of Pathology, University 
Medical College ; Pathologist to the Board of Health, New York 
City ; Visiting Physician Bellevue Hospital. 

Mr. B. T. Galloway, Chief of the Section of Vegetable 
Pathology, Department of Agriculture, Washington, D. C, 
and Editor of the Journal of Mycology. 

In the next number we expect to have our list completed 
and to have the honor of giving a short biography of each of 
our collaborators. 



NOTICE. 



We refer our readers and all interested, to our club rates 
with other excellent publications. See index. 



64 THE BACTERIOLOGICAL WOULD. 

LESSORS OJS T BACTERIOLOGY. 

( Copyrighted.') 



BY PAUL PAQUIN, M. D., V. M., 

Missouri State University. 



LITERATURE. . , 

Cornil & Babes — Les Bacteries. 

Prudden — The Story of Bacteria. 

Prudden — Dust and its Dangers. 

Thome — Structural and Physiological Botany. 

Mills — Comparative Physiology. 

Bell — Comparative Anatomy and Physiology/ 

LESSON SECOND.— PART I. 

THE CELL. 

In some sense the cells are to organized bodies what the 
atoms are to unorganized bodies ; namely, each are the con- 
structive elements of the whole, and the energy proceeds from, 
them. In chemistry, the atom is inferred, so to speak ; it 
cannot be seen ; but in histology the cell may be seen with 
magnifying instruments, and its size and shape examined, its 
life appreciated. 

When an anatomist dissects a complex body, the tissues of 
a man, for instance, he finds a bony framework surrounded 
by muscles, themselves incased in tegument, and the whole 
provided with vessels, nerves, etc. He finds, besides, lodged 
in the several cavities, various organs obviously meant for 
different purposes. All these portions of the body, with such 
a variety of aspects, consistence and shapes, appear as though 
formed by very different elements. 

The anatomist may search deeper, with much skill, in the 
tissues composing these different organs, and separate every fibre 
as far as possible with the naked eye, but he will fail to decipher 
the nature of the marvelous structure. 

If, however, a microscopical investigation is made in the 
tissues by proper methods, the darkness gradually disappears, 
the obscure veil is lifted, and the vital elementary base of the 
admirable composition of the body of man stands revealed. It 



THE BACTERIOLOGICAL TTOELD. 65 

is found then, that, notwithstanding the apparently very 
different nature of the several portions, they all are formed of 
cells. 

The white, compact, bony framework ; the pink, soft 
muscles ; the nerves ; the solid portions of the blood and lymph, 
all are made of cells. Thus, a complex body is simply an 
agglomeration of cells, which have been generated since the 
moment of conception, and gradually modified during their 
growth, to suit the needs of the different organs of the. 
individual, until they have acquired the forms in which we find 
them in the tissues of the adult stage. The first cell was the 
ovum, at the expense of which were generated new ones ; these 
in their turn generated new ones, and so on, and so on. 

From a typical cell usually of a definite, globular form as 
the ovum is, and the no less definite, rounded, primitive 
embryonic cells as exist in the embryo, we get, by process of 
evolution, different groupings of cells of various shapes, forming 
the several tissues which differ so materially in aspect and 
properties. The labor has been " specialized," * and divided 
among the innumerable cells, forming the whole structure. 

A muscle has the special function of performing various 
movements more or less rapid and more or less powerful ; its 
cells, therefore, are elongated and so endowed that they may 
cause energetic contraction, and relaxation of this organ. The 
red blood cells are loose, floating in the plasma of the blood, 
and individually have the power of absorbing oxygen in the 
lungs, which they afterwards convey to every portion of the 
body. The white blood corpuscles or leucocytes are also floating 
in the lymph and blood, wandering even through the tissues, 
and have the power of reproduction to such a degree, as to be 
capable to even produce new tissues, to fill the gaps made in the 
living body by wounds or disease. 

We see, then, that a man's body is composed of cells, 
modified in shape, appearance and properties, and fitted for 
their special respective functions, or for the special functions of 
the organ, or fluids, of which they are the component parts. 
They are individually endowed with life/ 



M. Prudden. 



66 THE BACTERIOLOGICAL WORLD. 

As men in the construction of an edifice are divided, singly 
and in sets, for the purpose of performing different labors simul- 
taneously, to erect a building more or less imposing, so are the 
cells of living things separated, or grouped, for the purpose of 
erecting and sustaining the living edifice. 

Among the builders of a home, there are those who carry 
bricks to the wall, those who carry the mortar, those who do 
the woodwork, etc., etc., and foremen supervising and directing 
the whole force. 

Among the cells building a complex organism there are 
the blood cells carrying oxygen to the tissues ; there are the 
muscle cells building the soft muscular structure, etc., etc., 
and there are the cells of the nervous tissue, supervising and 
directing, as it were, the whole force. And, somewhat as 
specialists in all vocations become experts in their line, and 
unfit after a time for any other labor than their specialty, so do 
the living cells, as I have just stated, become, by evolution, 
specialists unfit for duties other than their respective specialties. 

The bone cell, for instance, has lost the power of reproduc- 
tion whilst the lymph cell has retained it., The muscle cell has 
developed remarkably, to the exclusion of others, the power of 
motion, whilst the nervous cell has become endowed with 
mental energy and sensibility of the most wonderful character. 

The same cellular foundation underlies all complex living 
things, whether of the animal or vegetable kingdom. 

We gather, then, from what precedes in this lesson, that, 
whilst a complex living being of whatever reign or species, 
is endowed as a -whole with diverse properties more or less 
pronounced, it owes all to the different special properties, and 
the activity of numberless cells, all working co-ordinately for 
their own lives and the benefit of the individual. 

We see that the minute living organisms composing the 
body of an animal, or the trunk of a tree, are joined together 
for an important object, i. e., the propagation of the species, 
and it will be apparent too, that, at the outset, the primitive, 
component cells of the individual must be beings of a low 
organization. The ovum, indeed, is not unlike some little 
animal living masses, unicellular beings of the ponds, — the 



THE BACTERIOLOGICAL WORLD. 67 

amoeba, — which never develop beyond their first natural condi- 
tion, and propagate themselves by the reproduction of new, 
independent, separate cells. Some of the cells of the animal 
body retain this low organization until the death of the 
individual. The leucocyte, whose function we have seen and 
which is so essential to the life of the whole, remains much as 
an amoeba, and moves in the liquids of the body as the amoeba 
moves in water. This cell has retained its power of reproduc- 
tion as already stated ; when a breach occurs in the tissues 
it groups its successive progenies together, and fills it with 
tissue formed by them. 

In the development of a complex being, then, the principle 
involved is the grouping of innumerable, individual, successive 
cells, originally from a common first parent, the ovum, each 
having life independently of the other ; the relation existing 
between the largest and most complex bodies, and the smallest 
and most insignificant forms, lies in their common cellular basis. 
A man is related to the microscopic, unicellular, jelly-like mass 
known as the amaiba inasmuch as he himself originated from 
such a mass ; his body is the product of the like, and in the 
canals supplying it with nourishing fluids float constantly 
certain lowly cells with amoeboid movement. 

If we want to trace the relationship more closely, let us 
descend from man down to the most minute being (or vice versa), 
and examine as we proceed the various intermediate forms of 
the downward scale (or upward scale). 



68 



THE BACTERIOLOGICAL WORLD. 



*«V£ 



1®>9 

a 



& 




*^^^_ 



Figure 1, A series of vegetable cells and tissues and a series of 
animal cells and tissues, showing unicellular organisms as well as 
cellular structures of higher beings of both kingdoms. First row, a 
yeast plant ( torula cerevisise ) one-cell vegetable mentioned in the 
text ; b, common green mould ( penicilium glaucum ) , widely known 
microscopic plant composed of but few cells ; c, section of fig tissue, 
showing cellular arrangement ; d, longitudinal section of maple 
wood showing arrangement of fibres made from cells. 

Second row, a', amoeba, a common microscopical, unicellular, jelly- 
like animal resembling the white blood corpuscle ; b', fresh water 
polyp (hydra viridis), animal made of a little sac of jelly composed 
of comparatively few cells ; &, a section of cartilage tissue of an ani- 
mal showing disposition and appearance of cells ; d', section of intes- 
tinal muscle showing the grouping of fibres constructed by the cells. 

( Borrowed from Wyeth, Bell, Thome, Mills.) 

From the highest being to the lowest it is only a question 
of cells. The highest is constituted by a more perfect com- 
munity of cells, among whom the division of labor has been 
such as to produce particular, pronounced skill for particular 
functions ; the lowest is a single, independent being doing by 
itself everything necessary for its existence, but doing " noth- 
ing particularly well except reproduction." As a consequence 
the higher the organism, and the better its component cellular 
bodies have been trained, specialized by evolutionary influences, 
the more perfect is the whole living system ; and, the lower the 



THE BACTERIOLOGICAL WORLD. 69 

organism, the more imperfectly it performs the acts necessary 
for its existence. 

Life in a complex body does not exist as a unit as it were, 
for the whole individual, but it exists in the protoplasm of each 
and every cell forming it. 

The classes of living things, then, considered from a struc- 
tural point of view are only two : The unicellular organisms 
and the multicellular organisms. This is true of the vegetable 
and animal kingdoms. ( See Figure 1.) 

A plant may be formed by either only one cell as the yeast 
plant (Figure 1, a, or torula saccharomyces cerevisise) which 
consists of an oval cell about 1-3000 of an inch in diameter, 
containing a semifluid granular protoplasm, surrounded by a 
transparent envelope, and which reproduces by fission or bud- 
ding, — each severed portion, or bud, becoming finally a new, 
independent yeast plant ; or a plant may be formed of only a 
few cells, as the penicilium glaucum ( Figure 1, b ) ; or it 
may be an agglomeration of a variety of innumerable cells as 
in any highly organized plant ( Figure 1, c and d). 

On the other hand an animal may consist also of only one 
cell as the amoeba ( Figure 1, a ), which is a little mass of 
nucleated protoplasm that reproduces by fission and occasion- 
ally by budding, thus forming separate new individuals-; or 
it may be a little higher in the zoological scale, and be formed 
of a comparatively small number of cells as the polyp, a green- 
ish or reddish animal about K of an inch in diameter, attached 
to weeds in water, constituted by a kind of sac formed by an 
internal and an external layer of cells, throwing out tentacles, 
and yet capable of contracting itself into a globular mass 
( Figure 1, V ) ; or it may be an aggregation of modified cells 
as in more complex and highly organized subjects, large 
animals and man. (Figure 1, c and d'.) 

But there are beings so small, and whose properties are so 
difficult to appreciate, that it is practically impossible to dis- 
tinguish to what reign, they belong, solely by the aid of the 
microscope. 

Early in this century, Bory de St. Vincent, and later 
Haeckel, created a new intermediary kingdom in order to over- 
come the difficulty : the protista, implying that this class of 
beings includes the first animals that appeared on the earth. 



70 THE BACTERIOLOGICAL WORLD. 

This new kingdom, whether it has or has not reason to be, 
remains to be determined. Certain it is that the exact bound- 
ary between vegetable and animal kingdom is not absolutely 
demonstrated among the lower organisms. We find, often, 
micro-organisms whose place in the two old kingdoms is diffi- 
cult to establish. It is among this class of beings that we find 
the microbes ( schizomycetes, bacteria, etc.). 

True, some high authorities have committed themselves, 
and placed most of bacteria in the vegetable kingdom, appar- 
ently with sound reason, but still the evidence is not convinc- 
ing in all cases, though it seems to be in most of them. It is 
well to remember, too, that here, perhaps, lies the key to the 
solution of the relation between the cells of the vegetable and 
animal kingdoms. Indeed, when we descend from the highest 
vegetable to the microbe, and the highest animal to the microbe, 
we come to a focus, and meet at one point an individual that 
often seems either or neither, as in the following diagram. 

This long story about the cell may seem a superfluous 
digression. It is in fact a digression, but its utility is apparent 
in showing the relation of all living beings, through their cells, 
and it will be more appreciated further when we proceed to 
study the relation of microbes to tissues. 

PART II. 

SCHIZOMYCETES IN" GENERAL. 

Distribution. — At the bottom of the vegetable scale we 
find the algae and the fungi now known under the common 
term of thallophytes. The algae and the schizomycetes form 
a parallel series of beings, but the former usually contain 
chlorophyl, and the latter usually do not. The algae, 
therefore, are capable of preparing, manufacturing in fact, 
like other plants provided with chlorophyl, the nutrition 
necessary for their development. The schizomycetes, on the 
other hand, can live only at the expense of already constituted 
organic matter which they absorb in causing in them special 
transformations — putrefaction, fermentation. 

We find them everywhere in nature, and their power of 
wonderfully rapid reproduction by fission, and spore formation 
in many kinds, counterbalances their minuteness. 



THE BACTERIOLOGICAL WORLD. 71 

They exist in the ponds, creeks, rivers, lakes, the sea. 
They are* on and in the surface soil and the manure heaps. 
They float accidentally in the air, and thence fall on infusions 
of vegetables, dead vegetable and animal bodies, fermentable 
juices, as cider, beer, wine. They attack the milk, cheese, 
butter, lard, meats and other foods. 

They penetrate into the nasal cavities, the lungs, the 
intestines, and may enter the circulation and locate in the liv- 
ing tissues. Indeed, in the normal state, the skin, mouth, and 
intestinal excretions of man and beast, contain microbes 
innumerable. Through the mucous membranes, abrasions and 
natural ducts, some of them may invade the organism, and 
cause the most serious complications in the living machine. 

Miquel, in making experiments to ascertain the relative 
quantity of germs in the air, found that there were 5,500 per 
cubic metre in the Rue de Rivoli ; 11,000 in the operating room 
of the Hospital la Pitie, Paris. At an altitude of 2,000 metres 
there was none. After a rain the air was washed and com- 
paratively free from microbes. 

In water he found as follows : 248,000 per litre of rain 
water; 4,800,000 per litre of river water (water of the river 
Seine passing through Paris ). 

The proportion found in the air of the streets of Paris is 
startling and seems extremely high. Experiments made in 
other parts of the world have shown a much smaller quantity. 
Prudden, in his exceedingly interesting little book entitled, 
"Dust and its Dangers," records the average number of 
bacteria found in various parts of New York City as 376 per 10 
litres ( i. e. } in a volume about equal to a cube 8 inches square ). 

Carnelly found in the town of Dundee, Scotland, from 10 
bacteria to 170 in ten litres of air. 

It is self-evident that various atmospheric and telluric 
conditions will vary the quantity greatly. In dry, dusty 
weather the winds will raise a perfect shower of germs with the 
dust of the streets and roads, etc. This is particularly true 
when dry street sweeping is practiced. In rainy and snowy 
weather the germs will be brought to the earth in variable 
numbers. Again they are comparatively scarce in high alti- 
tudes. 



72 THE BACTERIOLOGICAL WORLD. 

In the water, it is needless to say, that surface water, 
shallow lakes and ponds, and the rivers receiving the city 
sewages and the draining of the shores, are more or less laden, 
according to the greater or smaller amount of nutritive organic 
matter that they contain, — being consequently far greater in 
the immediate neighborhood than at long distances from 
populous centers. 

Indoors, the number of germs floating or deposited is com- 
paratively greater than outdoors, particularly in houses imper- 
fectly ventilated by chance apertures or chimneys. The 
number found floating in the air of a room depends on the 
stillness or stirring therein. For instance at midnight, when 
all has been still for hours, the minimum number is found, 
whilst in the morning, after making up the beds, for instance, 
causing the visible and invisible dusts to fly from the fabrics, 
clothes, furniture and floor, where they had sunk during the 
night, the number is incomparably greater. Then again there 
is a vast difference before and after sweeping. 

Tucker, in testing the number of germs in the wards of the 
Boston City Hospital, found by the hourly examination from 
the still hour of midnight to late in the stirring hour of the 
day, 1 to 477 bacteria per 10 litres of air. 

Oarnelly found in clean houses 180 bacteria per 10 litres, 
and in dirty houses 900. In dirty schoolrooms with common 
ventilation the same writer found 2,000 living bacteria in the 
same volume of air, whilst in clean schoolrooms, mechanically 
ventilated, he found from 30 to 300. 

Prudden exposed five minutes a dish of 3% inches in 
diameter, in a carpeted living room in New York, before 
sweeping, and found that 75 bacteria had settled onto the sur- 
face. He repeated the experiment when all was still, immed- 
iately after sweeping, and found 2,700 bacteria. It is needless 
to proceed further on this line, to point out the danger lurking 
from germ-laden air or water. 

Yet, we must not think that all these germs are harmful. 
Indeed, comparatively few are, and even these do not always 
cause injury to animal life, because, in the first place, they do 
not always find in the animal bodies in or on which they settle 
means of ingress into the organism ; in the second place, the 



THE BACTERIOLOGICAL WORLD. 73 

tissues and liquids do not present always suitable nourishing 
media, particularly in the normal state; in the third place, 
the cells composing the higher organized beings are endowed 
with life, activity, energetic physical resistance, and being- 
defended by the leucocytes, who seem to have the power of 
overcoming invading bacteria by phagocytism,* offer an 
effective barrier to the microbes ; finally, the blood serum seems 
to be germicide. 

But, on the other hand, there may be among the water and 
air bacteria, some so endowed and constituted, that their 
presence in the skin, in the mouth, the nasal cavities, the 
lungs, the intestinal tract, is a constant menace to health. 
And, to individuals presenting some pathological conditions, 
some germs are a positive danger. Note for instance the 
exposure of a man, suffering with a bronchial catarrh, to the 
dust of a hospital ward, or the dust of a bedroom containing 
tuberculous patients ; note, again, the exposure of a wounded 
individual with a skin abrasion to the atmosphere of a badly 
ventilated, dirty house ! In the former case tuberculosis may 
be the result, because the lungs present to the tubercle baccilli 
a proper, sticky nidus for their adherence, vegetation and 
multiplication, and the tissues of the body are perhaps weakened 
by the catarrhal affection. Healthy subjects, however, inhal- 
ing the same air, would likely suffer nothing thereby. In the 
second case, septicemia may occur, by the action in the wound, 
of germs always present in the intestinal tract, on the body and 
in the air, etc., but which succeed in overcoming the organism, 
only after gaining a footing in or through a weakened spot of 
the tissues, such for instance as are produced by cut wounds, 
or contused wounds with abrasion. 

On the other hand, there may float accidentally in the air, 
or exist in water, a few extremely dangerous bacteria, such as the 
germs of Asiatic cholera, yellow fever, diphtheria, small-pox, 
glanders, charbon, which determine these special diseases, often 
in epidemic form, seemingly irrespective of the condition 
of health of the susceptible beings. 



* The leucocytes, white blood cells, are termed phagocytes because 
they are believed to absorb and digest bacteria. 



74 



THE BACTERIOLOGICAL WOELD. 



Form and classification of schizom ycetes.— As we have 
already seen more than once, the schizomycetes are unicellular 
beings of a very minute kind, varied in appearance, ranging 
between one-ten-thousandth of a millimetre and a few thou- 
sandths of a millimetre, (Fig. 2.) Without attempting per- 
haps to classify them fully, Oohn has divided the bacteria, 
according to their shapes, into four groups : 

First. — Spherobacteria, or the globular forms ( commonly 
known as cocci ). 

Second. — Microbacteria, short rod forms ( bacterium). 

Third. — Desmobacteria, long rod forms ( bacilli ). 

Fourth. — Spirobacteria, spiral forms. 

This grouping might serve in a degree as a classification, 
but the forms are not constant during all the stages of microbic 
life. 

Several authoritative naturalists have attempted to make 
satisfactory classifications, but none of these are free from 
objection, because of the still limited knowledge of a positive 
character, possessed by scientists, on the morphology of these 
infinitely small beings. 




Figure 2. A variety of microbes showing the different forms 
constituting the basis of classifications. ( Borrowed partly from 
Cornil and Babes.) 

At the present writing, it appears thjat the following 
classes have the greatest number of followers. They were 
proposed by Van Tieghem in 1884, who places all the schizo- 
mycetes in the family of bacteriacees. 



THE BACTERIOLOGICAL WORLD. 75 

The class known under the name of : 

Micrococcus, or coccus, is constituted by the individuals 
composed each of little round cells. 

Bacterium, cylindrical cell. 

Bacillus, cell of the rod shape. 

Beptothrix, the indefinitely long filament without encasing 
sheath. 

Crenothrix, the same kind of filament but provided with 
encasing sheath. 

Cladotlirix, sheathed filament with ramifications. 

'Vibrio, coiled filaments that may be dissociated. 

Spirillum, long, spiral, screw-shaped filament (longer than 
vibrio ) . 

Spirochete, still longer spiral filament with numerous 
coils. 

Be Bary, from a more scientific basis, classifies the microbes 
called bacterium in the above, under the head of arthrobac- 
terium and bacillus. 

Now it often happens that the microbes of some of these 
species are grouped together in various ways. These special 
aggregations have been named by Van Tieghem as follows : 
The masses of cocci ( zoogloea ) united by a thick gelatinous 
substance are termed ascococcus ; adhering together, punctula ; 
when cylindrical cells ( bacterium ) are united by gelatine they 
take the name of ascobacteria ; without gelatine polybacteria ; 
united masses of rod forms or spirals are named myconostoc. 

Another convenient classification is that of Winter and 
Flugge, which I give in tabulated form : 

GENERA. 

( Isolated, in chaplets or in zoogloea? . . . . . . . Micrococcus. 

( In large numbers 
Round and 

or j ( Solid colonies | irregular colonies .. .. Ascococcus. 

oval ) filled J 

cells. Forming ] with cells. ) In small definite 
zoogloeae j numbers 

in the i and 

form of t regular groups .. .. Sarcina. 

I A single circular layer Clathrocystis . 



76 



THE BACTERIOLOGICAL WOKLD. 



f Short, isolated, in a mass or in zoogloeae 



.9 



Forming 
long 
^ filaments. 



( Isolated, 
inter- 
laced 
or in 
bundles. 



r& f Straight 
,2 filaments. 



( Short, jointed 



Bacterium . 
Bacillus. 



Longim- ( Slender Leptothrix. 
. perfectly < 
^ jointed. ( Thick Beggiatoa. 



Spiral 
filaments 



Short, rigid 
Long, flexible 



l^With false ramifications 



Spirillum (Vibrio). 

Spirochete. 
I Streptothrix . 
{ Cladothrix. 

Myconostoc . 



\^ In zoogloeae . . 

When the micrococci or cocci are two by two they are 
termed diplococci ; when in strings or beads, streptococci. 
The cocci are about of the average of Ojn 2.* They are the 
smallest organisms that can be seen with our present optical 
instruments. 

The various rod-shaped and spiral micro-organisms vary in 
length between 1 and 7 or 10ju and in thickness between 1 and 2/t. 

Variation in form. — The classification of the microbes 
rests, as we see, chiefly on the forms under which they present 
themselves, in the period accepted as the adult stage, but, 
as already mentioned, we must not infer that the shapes forming 
the basis of these classes are constant during the development 
and life of the microbes. On the contrary, certain microbes 
appear in diverse forms during their development, and the sizes, 
even, may be influenced by the nutritive surroundings. Fig. 3. 



*The u, mikron, a ,unit in certain microscopic measurement, 
means micromillimetre, and is equal to one-thousand fch of a millimetre. 
A millimetre is, as the word implies, the one-thousandth of a meter, and 
a meter is equal to 39.37 inches. A millimetre, then, is nearly 1-25 part 
of an inch and ju ( mikron ) is the 1-1000 part of 1-25 of an inch. 



THE BACTERIOLOGICAL WORLD. 



77 



If I 1 



© /a ^ 




Figure 3, Bacillus Mag at er him, showing various appearances 
during growth : a, outline of a motile chain of rods in active vegeta- 
tion ; b, a pair of motile rods in active vegetation ; p, a quadricellular 
rod in a state of b after treatment with alcoholic solution of iodine ; 
c, a five-celled rod in the first preparation for forming spores ; d — ■/, 
successive stages of a pair of rods while forming spores ; d, about two 
o'clock in the afternoon, e, about one hour later, /, an hour later than e. 
The spores in formation in /are ripe towards evening ; no others were 
formed ; the one which apparently began to be formed in the third 
cell from the top in d and e has disappeared ; the cells in /, which did 
not contain spores, perished by about nine o'clock in the evening ; 
r, a quadricellular rod with ripe spores; gi, a five-celled rod with 
three ripe spores placed in a nutrient solution after several days' desic- 
cation, half an hour after noon ; g 2 , the same specimen at about half 
past one; g3, the same about four o'clock; 7ii, two spores with the 
walls of the mother-cells dried and then placed in a nutrient solution 
about forty-five minutes after eleven ; h 2 , the same about half past 
twelve ; i, k, Z, later stages of germination as explained in the text ; m, 
a rod formed from a spore placed eight hours before in the nutrient 
fluid and in the act of splitting transversely ; a mag. 250, the rest of 
the figures 600 times. ( Borrowed from De Bary.) 

Generation. — From this point of view, Van Tieghem 
divides the schizomycetes into endosporous, meaning those 
capable of reproduction by spore formation, and arthrosporous, 
those in which spores are not known to ever exist. The usual 
mode of reproduction, however, is by fission, that is to say, 



78 . THE BACTERIOLOGICAL WORLD. 

successive divisions or bi-partitions, each cell dividing into " two 
daughter cells through an unlimited number of generations. " 
As a rule, the division occurs in one direction making the 
successive cells appear all in one row, as a long, segmented 
filament, providing that no serious interference has been 
interposed on the spot at the time of nutrition and growth. 
The microbes, in process of dividing thus in one direction, 
stretch out, a wall divides them in two, the extremities in 
contact become rounded and then the two parts separate. The 
diplococci and streptococci mentioned above are, doubtless, in 
most cases, germs thus joined together during the process of 
growth and segmentation. 

In long spiral forms, the multiplication may occur by the 
segmentation of the whole into several short rods, each keeping 
in a measure a wavy appearance derived from the spiral shape 
which they gave to the organism before separation. 

In the crenothrix, the reproduction occurs by both vertical 
and transverse segmentation at the terminal point of the 
filaments, that is to say, the cell divides at that part into four 
new cells, having first the appearance of little angular bodies, 
but the angles of which soon become blunt until the new beings 
are fully rounded. Then they grow into filaments if properly 
nourished. 

In certain round cocci the division occurs in various 
directions. 

In the sarcina, a class of microbes described in Winter, 
Eadenhorst and Flugge's classification, division occurs in three 
ways through the organism,, thus forming little cubical masses. 

In some germs of the air, reproduction occurs by division 
within a large capsule. The sporogenous microbes belong 
chiefly, so far as now known, to the bacilli and a few other 
filamentous forms. Each cell usually generates but one spore, 
rarely two. Some authors have described bacteria throwing off 
numerous spores ; it is possible that in such cases the separating 
walls between the spores passed unnoticed. 

The first thing noticed in the process of spore formation is 
the changes in the previously homogeneous protoplasm ; it 
gradually becomes darker, and even granular, sometimes giving 
the amyloid reaction. Soon a darker, minute body is seen 



THE BACTERIOLOGICAL WORLD. 79 

within the envelope of the mother-cell ; it acquires distinct 
contour, grows in size and reaches its largest volume in a few 
hours. This is the spore. It now is strongly refringent, of a 
colorless or slightly bluish, glistening appearance. At this 
time, the protoplasm of the mother-cell, which has been 
gradually utilized to make the spore, has about completely 
disappeared from around it, its envelope may break to liberate 
the new body, or it remains, for a while, apparently as a 
disintegrated shell around the glistening mass. 

The size of spores varies between Opil and 0ja6, Shape, 
spherical or oblong. 

Structure and contents. — The schizomycetes all have a 
surrounding wall or membrane and contain a protoplasm. The 
analyses of Nencki show the chemical composition of both the 
membrane and content to be, usually, a special albumenoid 
substance named mycoprotein. However, there may be some 
other chemical agents entering into it. It seems certain that 
the envelope contains in certain cases some cellulose. Certain 
bacteria contain no mycoprotein but are constituted by other 
chemical matters resembling in some cases the caseine of plants 
and the mucous matter of animals. 

There is a tendency, in the envelope of several microbes, 
to form gelatine or to become gelatinous itself. In such cases 
we sometimes find a number of them imbedded in gelatin- 
ous masses (zoogloea, etc.), or if the gelatine is thrown around 
individual germs, as in the diplococci of pneumonia, they 
become encapsuled. The membrane is sometimes naturally 
colored yellow, red or blue. 

The homogeneous protoplasm of the cell is more refringent 
than water. It sometimes contains small ^specks or granules 
which perhaps are of a fatty character; vacuoles (or empty 
spaces) are exceedingly rare. Iodine stains the protoplasm 
yellow. 

Both the envelope and protoplasm have a marked resistance 
to bases and acids which destroy animal cells. Some authors 
have seen in this an indication pointing to the vegetable 
character of bacteria. 



80 THE BACTERIOLOGICAL WORLD. 

It is also on this resistance to reagents, and the germ's 
affinity for aniline dyes, that the coloring methods of bacteri- 
ology are based. Iodine reagents, by the blue color that 
resulted in the protoplasm of certain microbic cells, made 
possible the discovery of amyloid substances. 

Motility. — Certain schizomycetes are endowed with the 
power of moving and others are not. This property is due to 
vibratile cilia at the extremities of the bacteria, the filamentous 
forms having two to six whilst, according to Zopf, the round 
germs have only one. Only Ehrenberg and Cohn seem to have 
positively identified cilia in the cocoid forms. On account of 
their constant motion, and of their refringence being so near 
that of water, it is extremely difficult to observe the cilia of the 
fine germs of this character. 

The non-motile germs have no cilia. Certain bacteria 
grow cilia only in case of necessity, such as when they proceed 
from the bottom of a liquid to the surface for the purpose of 
nourishment. 

The movements of bacteria are very interesting and rapid, 
and should not be confounded with the well-known brownien 
movement. The movements of motile microbes are incessant 
and in all directions, whilst the molecular brownien movement 
which is often present also in certain bacteria, particularly the 
spherical forms, is a characteristic trembling, or whirling, on 
the spot, that the eye soon learns to diagnose. These motions 
are, perhaps, due to the shape and composition. 

[Lesson third in March number.] 



THE BACTERIOLOGICAL WORLD. 81 

SPECIAL TRANSLATIONS FROM THE 

GERMAN. 



[BY H. W. LOEB, A. M., M. D., ST. LOUIS, MO.] 



THE ROMAN MALARIAL FEVER, ESPECIALLY IN SUMMER 

AND AUTUMN. 

By A. Celli and E. Marchiafava, 
Of Rome, Italy, in the Berliner Klinische Wochenschrift, Nov. 3, 1890. 

In order to obtain a proper understanding of the before- 
mentioned change of the blood by malarial infection, one must 
make, as we indicated some time ago, a large number of 
observations upon the severer and milder cases of malaria, both 
during the stage of apyrexia and the febrile stage. 

It is well known that malarial infection is produced 
by a specific parasite, which wanders into the red blood cells 
in which and upon which it lives and develops, changing 
further the haemoglobin into melanin and which multiplies by 
division. Besides, Golgi, who studied malarial fever at Pavia, 
has shown the connection between the different phases of the 
development of this same parasite and the course of the fever 
curve. He also discovered that the parasite of the quartan 
and of the tertian varieties differed both morphologically and 
biologically. We have busied ourselves with the summer and 
fall malarial fever of Rome, and we can state that in these the 
small amoeboid parasite exclusively is found. 

The profession knows that the malarial fever of summer 
and fall has a character entirely different from that of spring. 
The latter form has constantly a distinct cyclical course ( quo- 
tidian, tertian, etc.), is never pernicious, recovers of itself 
generally without quinine/ and is always cured by quinine. On 
the contrary the fever of summer and fall is not cyclical. The 
attack occurs in the last-named'form of malaria, as a rule, daily, 
and lasts, frequently, with trifling fluctuations of temperature, 
twenty-four, thirty-six or more hours so that fever and 
apyrexia imperceptibly merge into one another. In these 



82 THE BACTERIOLOGICAL WORLD. 

cases when the initial chill is missing, the apyrexia is of shorter 
duration and the headache and malaise persist for a long time ; 
the patients offer no clear distinction with reference to the 
type of. fever, and they maintain what now and then corresponds 
to the truth — that they have been attacked by a continued 
fever. These are the severe pernicious cases which seem 
preferably to attack the central nervous system. They never 
recover spontaneously, and quinine does not operate so certainly 
as in the milder cases. The frequent relapses are characteristic, 
as also severe attacks which occur as a sequence. Finally to 
these belong those cases of pernicious fever with trifling eleva- 
tion of temperature, even with subnormal temperature. In 
these cases, as we have published in a former article, the regular 
so-called small plasmodia ( Piccoli plasmodi ) occur. The red 
corpuscles contain at the time of the acme of the fever one or 
more living, moving parasites. The form is the same in the 
sweating stage and in the begining of the apyrexia. 




THE BACTERIOLOGICAL WORLD. 
2 3* 



83 







a 



9 



19 







// 



/2 



13 






16 



17 




18 




19 




20 




22 





2¥ 



US 





EXPLANATION OF THE DIAGRAMMATIC DRAWINGS. 

Figures 1-6. Circular form of the small pigmented Plasmodium. 
In figures 4 and 5, the sporulation is ended and no pigment appears. 

Figures 7-20. Circular form of the small pigmented plasraodiuin. 

Figure 7. Ring-form with a haemoglobin bud in the middle. 
( Hcimoglobinauge.) 

Figures 8-9. The haemoglobin bud is broken up in more divisions. 

Figure 10. Pigment form from the decomposed haemoglobin bud, 
(See 7-9.) 

Figures 11-12.- Amoeboid and non-moving form with its pigment. 

Figures 13-14. Motionless parasite with pigment in the center. 
In 14 the haemoglobin has encircled the parasite. 

Figures 15-20. Spore formation. In 15, the beginning. In 20 the 
pigment is surrounded by free s]x>res. 

Figures 21-22. Atrophic red blood cells with plasmodia. 

Figures 23-25. Young crescentic bodies — Laveran's parasites. 



84 THE BACTERIOLOGICAL WORLD. 

However the plasmodia show, some hours before the 
attack, a certain change so that one can prophesy the onset of 
an attack by the observation of the blood. 

Before the attack one may observe : 

First. Kound, motionless, small distinct bodies which 
contain in the center a haemoglobin flake or pigment grain. 
( The ring-shaped form.) Fig. 7-10. 

Second. Amoeboid also small, but more or less moving 
organisms with an indented contour and of a grayish white 
color. They contain the smallest pigment grains, visible only 
with the greatest care. Fig. 11-12. 

Third. Large, round, motionless, white parasites with 
pigment flakes situated in the center or at the periphery. The 
still remaining haemoglobin lies in concentric layers around the 
Plasmodium while the edges of the red blood cells have become 
paler. Fig. 13-14. 

All of these circular forms which we have just sketched, 
but especially figures 21 and 22 of the above drawings occur at 
the end of the apyrexia aud in the beginning of the fever 
attack. And indeed the parasites are found in the red blood 
cells, which have diminished one-third in size, are irregular, 
and which have the color of brass. After the entrance of the 
parasite the atrophied blood corpuscle is very easily, by a little 
practice, discriminated from such forms, which one observes 
under the microscope, in the decomposition of the blood. In 
the pernicious cases the parasites are seldom in the stage of 
sporulation, while they occur frequently in the quartan and 
tertian form, and especially in the beginning of the attack. 
These spore-carrying parasites occur exclusively in the red 
blood cells. However, only a portion of the latter remains, in 
the middle of which is visible a pigment flake, and more 
seldom a living, moving parasite or some pigment crystals. 
Fig, 15-20. 

Unpigmented plasmodia in the state of sporulation are 
formed only rarely in the blood taken by puncturing the finger, 
but are very numerous in the brain-capillaries, especially in 
pernicious fever. In the beginning of the paroxysm of fever, 
and for some hours succeeding, the unpigmented amoeboid 
forms only occur. 



THE BACTERIOLOGICAL WORLD. 85 

In distinction from the parasites of tertian and quartan 
fevers, which the eminent Golgi has studied, the plasmodia of 
the fall and winter fever of Rome can remain living and mov- 
ing, for a long time, free of pigment. When the motion 
ceases, they take on the ring-form. Then they contain haemo- 
globin flakes which in the form of a wreath enclose the center. 
This is the stage of sporulation • spore formation, however, 
only takes place within the red blood cells. When this occurs 
the blood cell is soon destroyed. The inquiry upon this prob- 
lematical stage of sporulation seems of special interest. In 
this, we do not, by examination of the blood-of the skin, obtain 
sufficient explanation ; we must search the blood of the inner 
organs, in life as well as after death. In this way we meet the 
curious fact that spore-carrying parasites may disappear for 
some time from the finger blood, while they are present in the 
blood of the spleen, obtained by aspiration during life. So it 
may happen that spore-carrying plasmodia may disappear from 
the living blood, while they are plentifully found in the blood 
of the inner organs especially in the brain. 

This opposes the objection that in these cases the develop- 
ment of the surviving parasites continues after the death of the 
host, because the blood of the veins of the arms, taken during 
the autopsy, is found free from spore-carrying parasites, while, 
in the blood of the brain, pigment and spore -carrying parasites 
occur in large quantities. Hence it follows that in the tertian 
and quartan fevers the entire development cycle of the parasites 
is visible in the blood of the finger, while in the autumn fever 
of Some all of the development stages do not appear in the 
same blood. Whereas spore-carrying parasites are not gener- 
ally found in the finger blood, it is however easy to foretell the 
approach of the fever paroxysm by examination of the blood. 
If the pigmented plasmodia here described are found, it can be 
foretold for the next few hours with all certainty. Our infor- 
mation hitherto may be reduced in the following way : 

The parasite of summer and autumn fever runs through 
its entire course, in a very short time. It may appear with 
this spore formation and without pigment formation. The 
quotidian and fever forms with short apyrexia agree in this. 
In these, too, the process of development from the amoeboid to 



86 THE BACTERIOLOGICAL WORLD. 

spore-carrying plasmodium is completed in twenty-four hours ; 
yes, in a shorter time. In these cases the spore formation may 
occur so quickly in the inner organs that the colony breaks 
down into daughter cells before they are carried in the blood- 
stream to the blood of the hand, The blood of the finger will 
then thoroughly show spore-carrying parasites only when they 
are formed in very great quantities. 

In conformity with this view, the splenic blood obtain by 
aspiration in a case of pernicious fever contained : 

First. Amoeboid unpigmented forms, as m the finger 
blood. 

Second. Non-moving plasmodia which are found in the 
red corpuscles and especially in degenerated ones (this form 
"was rare in the finger blood ). 

Third. Large, white, unpigmented cells. 

Fourth. White cells, some irregular dark- colored flakes 
which the red and white cells contained. 

Fifth. Round transparent cells of various size, with black 
quickly-moving contents. 

Sixth. Bound plasmodia within the red blood cell with a 
pigment flake in the center. 

Seventh. Spore -carrying parasites in the interior of the 
red blood cell. 

The small amoeboid forms generally disappear from the 
blood upon the administration of quinine. In some cases, 
however, two or three attacks occurred before a complete cure 
even when large doses of quinine per os or subcutaneously were 
given. 

LATERALS CRESCEXT -FORMED PARASITE. 

We have observed the crescent-formed parasites, dis- 
covered by Leveran, since 1885, and considered them a pecul- 
iar grade of development of the plasmodia. Oelli and Guarneri 
showed later (Riforma Medica, 1888) that the plasmodia of 
malaria may run through two different phases of development. 
They appear first as the amoeboid form, and later as sickle- 
shaped bodies.' The latter recall the gregaria and the coccidia 
which Metschnikofl calls the parasites of malaria. Grassi and 



THE BACTERIOLOGICAL WORLD. 87 

Seletti name the amoeboid form hsemamoeba and the crescent- 
shaped, leverania. The crescent-shaped are exceedingly poly- 
morphous and they likewise enter the red blood cells and there 
go through their cycle of development. Most of the remaining 
red blood cells which have been attacked by the parasite are 
clearly visible, while in other cases the round spores alone 
remain. Sometimes the blood cells become quickly pale, and 
only their margin still preserves the original color. In some 
cases the crescentic parasite presents the same color as the 
blood cell. A double-folded membrane which is not always 
clearly visible surrounds the plasmodium, its contents being 
equally hyalin. The poles stand out clearly, by coloring the 
granular contents, in which a nucleus is visible. The latter is 
occasionally visible in the uncolored specimen. The pigment 
is equally well deposited over the crescent ( Fig. 25 ) but some- 
times it is massed in a circle or with irregularity. It is gener- 
ally motionless but it is also found in the round and flagellated 
forms in active motion, which is generally limited to the 
vicinity of the nucleus. Once we saw an unpigmented 
flagellated parasite. 

The crescent-shaped and spindle-form parasites may change 
into the oval and circular forms. The former breaks into 
round yellowish cells of the same or different sizes. This 
occurrence has only an external resemblance to sporulation. 
While the spores which one may observe in the spleen -pulp and 
marrow of bones remain for several days unchanged, the cres- 
centic parasites are rapidly destroyed so that at most for a 
short time is its pigment store discoverable. By the use of 
dilute mineral acids, organic acids and dilute alkalies, the 
crescentic bodies are just as quickly destroyed as the other 
malarial parasites. 

Laveran has discovered the crescentic bodies in all forms 
of malaria. He thinks, however, that they are found by 
observation more frequently m the severe return forms than in 
the fresh cases. 

We have seldom seen these parasites in the summer cases 
but most frequently in pernicious and cachectic cases, especially 
in autumn. Again, we have found them in cases which were 
not yet treated with quinine, during the attack of fever but 



88 THE BACTERIOLOGICAL WORLD. 

continually coincident with the unpigmented amoeboid forms. 
We have further seen parasites which would indicate a transi- 
tion form between the crescentic bodies and the amoeboid 
Plasmodia.. We are inclined to consider the crescentic parasite 
as a later stage of development of the amoeboid forms, as we 
have regularly seen the latter in all cases during the fever 
while the former even in the severe cases may disappear for a 
time from the blood. These results are obtained from researches 
upon the living blood in the skin, lungs and spleen. 

Case 1. — In a case of summer malaria of the quotidian 
type the aspirated blood of the spleen, before the fever 
paroxysm contained amoeboid and round pigmented forms and 
yet not so plentifully as the finger blood. Near by were the 
parasites of spring malaria and the spore-carrying forms in 
small quantities and in larger quantities the brass-colored and 
white forms with blood corpuscles filled with melanin grains. 

Case 2. — In a second case with exceedingly infrequent 
paroxysms, the blood of the spleen and lungs contained during 
the apyrexia the crescentic forms, and during the attack the 
small amoeboid forms. The blood of the finger exhibited the 
same appearance, only in the latter there was found less black 
pigment than in the blood of the inner organs. 

Case 3. — Sixth return, cachexia. The blood of the finger 
and of the spleen contained during the paroxysms only the 
small amoeboid forms. Quinine was then administered. The 
fever diminished. Now we found the crescentic bodies. 

Case 4. — Malaria existed for four months. Several 
relapses undergone. In the blood of the finger isolated amoe- 
boid and a few crescentic bodies. The same found in the 
spleen blood, also more numerous pigment-containing and 
unpigmented white blood cells. Besides this, parasites with 
pigment in the center, prepared to form spores. 

Case 5. — In the blood of the finger, numerous amoeboid 
forms, a few of which in the state of sporulation ; numerous 
crescentic bodies. In the 'spleen blood spore-carrying Plas- 
modia, a few crescentic bodies. 



THE BACTERIOLOGICAL WORLD, 89 

Cases 6 and 7. — The living blood contained few crescent- 
shaped and many amoeboid bodies. In the blood of the organs 
were found many crescentic and few amoeboid forms. 

In conformity with Councilman the crescentic and these 
metamorphosed bodies were found in greater number in the 
spleen. This difference is not found in the other cases because 
the different forms of the parasites in the various, simultaneous, 
blood examinations are almost uniformly distributed. 

The sections sent by Dr. Bignami give the same result. 
In pernicious fever the blood of the spleen and marrow of 
bones contained very numerous crescentic parasites butjthese 
are free of spores. In opposition to this numerous spore- 
carrying forms are found in the brain and lung capillaries. In 
support of the derivation of the amoeboid forms from the 
crescentic bodies, we have the inoculation experiment carried 
on by Gualdi and Antolisei. The inoculated blood contains 
only the crescentic bodies. The amoeboid forms alone, however, 
are observed in the blood of the inoculated patients at the 
beginning of the fever. However, if the amoeboid forms have 
been before observed in the blood of the patient who has been 
inoculated, and if they appear two days after the inoculation, 
the experiment presents no full evidence. It must be repeated 
with far more stringent cautions. 

A further proof of the derivation of the crescentic bodies 
from the amoeboid form lies in this that the former appears 
during the apyrexia and the latter during the fever. Of course 
this conclusion can only be supported by experimentation with 
a constantly small quantity of blood. Leveran^s and our own 
observation show that the crescentic bodies occur in the most 
varied fever types, that they are found simultaneously with the 
oval, round and spore-canring forms during the fever unless 
quinine has been administered. We saw these parasites — in 
conformity with Laveran — very often after repeated attacks 
and in the long-lasting fever heretofore called ansemia. We 
have also seen cases of pernicious fever without the crescentic 
bodies, some,- too, in which the parasites appeared first before 
the symptoms of pernicious fever had declined. 

All our observations indicate that the cases of malarial 
fever which occur in Eome during some years, especially in 



90 THE BACTERIOLOGICAL WORLD. 

summer and autumn, are characterized by a tendency to 
become pernicious on account of the daily repeated attacks ; 
that in these cases the small amoeboid parasites occur in the 
blood-corpuscles. These plasmodia have either a short devel- 
opment and are then unpigmented or they have a longer one 
when they contain pigment. The same thing occurs, accord- 
ing to G-olgi in the tertian, quartan and mixed forms. These 
parasites are found only in certain neighborhoods and in the 
light forms of malaria. Different malarial parasites corre- 
spond to the different seasons, the different malarial localities, 
and finally to the mild or severe forms of fever. 



WHAT MEASURES ARE DEMANDED AGAINST THE SPREAD 

OF DIPHTHERIA. 

Prof. F. Loeffler, of Greifswald, thus concluded his paper 
upon the above subject {Berliner Klinische Wochenschrift, 
October 6, 1890) read before the Tenth International Congress 
at Berlin : 

First. The cause of diphtheria is the bacillus diphtheri- 
ticus. It is found in the exudation from the diseased mucous 
membrane. 

Second. With the exudation is the bacillus scattered 
externally. It can be deposited upon everything which is in 
the vicinity of the patient. 

Third. Those who have diphtheria harbor these infection- 
producing bacilli as long as the slightest trace of the condition 
remains, and also some days after the disappearance of the' 
disease. 

Fourth. Diphtheria patients should be isolated so long as 
they still possess bacilli in their secretions. Children who have 
had diphtheria should be kept from school at least four weeks. 

Fifth. The bacillus diphtherias lives in the little pieces of 
membrane in a dry state for four or five months. It is, 
therefore, necessary that everything which can come in 
contact with the discharges of the diphtheritic patients, the 
wash, bedclothing, drinking and eating utensils, attendants' 
clothes, etc., should be disinfected by boiling in water or by 
treating with steam at a temperature of 100o C. ( 212o F.). 



THE BACTERIOLOGICAL WORLD. 91 

Further, the room in which the patient has been lying should 
be carefully disinfected. The floor should be repeatedly scoured 
with a warm, sublimate solution ( 1 to 1000 ), and the walls and 
pieces of furniture thoroughly rubbed. 

Sixth. The researches upon the vitality of the diphtheria 
bacillus in the moist state are not yet completed. The bacilli 
are, if possible, longer viable under a moist condition than in 
the dry state. Moist, dark dwellings seem, indeed, to be propi- 
tious for the preservation of the diphtheritic virus. Such 
dwellings should, therefore, be made hygienic ; especially is it 
necessary to care for the thorough drying of such places and 
for the entrance of light and air. It is especially necessary to 
observe caution in a change of residence that there has been a 
thorough disinfection of the formerly infected dwelling. 

Seventh. The diphtheria bacillus thrives outside of the 
body at a temperature of 20° C. ( 68° F. ). They grow very well 
in milk. It is, therefore, proper to oversee the milk business. 
The selling of milk from a farm-house in which diphtheria 
patients exist should be forbidden. 

Eighth. The diphtheria-resembling diseases of numerous 
animal species, of pigeons, chickens, calves, hogs, are not con- 
tracted from the bacillus of human diphtheria. The diphtheritic 
animal diseases are, therefore, not to be feared as a source of 
diphtheria. 

Ninth, The statements of Klein in regard to the etiological 
identity of the disease in cats, as observed by him, with human 
diphtheria are not yet certain and need wider investigation. 

Tenth. Lesions of the mucous membrane favor the fixation 
of the diphtheritic virus. Susceptible individuals, however, 
can become diseased without existing lesions. 

Eleventh. During the time in which diphtheria prevails, 
the cleanliness of the mouth, nose and throat of children should 
receive special care. Moreover, it seems advisable that the 
children should be instructed to use prophylactic mouth- 
washes and gargles with aromatic waters or weak' solutions of 
corrosive sublimate ( 1 to 10000 ), 

Tioelfth. The favorable influence of certain meteorological 
factors upon the spread of diphtheria is as yet not with 
positiveness known. 



92 THE BACTERIOLOGICAL WORLD. 

THE PATHOGENESIS OF TETANUS. 

Faber and Knud, in the Berliner l£linische Wocliensclirift, 
have succeeded in obtaining a filtrate, completely devoid of 
bacteria, from a strong, virulent tetanus-culture, which, inocu- 
lated into animals, reproduces positively the known symptoms 
of experimental tetanus. The poison operates after a certain 
period of incubation, which depends on the virulence and 
quantity of the filtrate. It acts by inoculation into the tissues, 
and also by intravenous injections ; but while in the former 
both local and general convulsions occur, in the latter the local 
convulsions do not appear. ~No immunity is obtained by the 
filtrate. Heating the solution for five minutes to 65 degrees 
causes every trace of the poison to disappear. — Centralb. fur 
Bakt. unci Parasit, Dec. 4, 1890. 



SPECIAL TRANSLATIONS FROM THE 

FRENCH. 



THE FILTRATION OF WATER. 



BY N. E. DUCLAUX. 

The study of microbes has completely transformed the idea 
that we had formerly on filtration. We asked, until of late 
years, that a filter only free the water of its matters in suspen- 
sion, to render it clear when it is cloudy or even only shaded, 
and every filter that accomplished this was declared good. We 
have asked, besides, afterwards, that filtration deprive the 
water of some of its elements in solution, for instance, 
the organic matters which give it a taste, and may render it 
impotable. All the filters do not give this result. The filters 
of charcoal are more apt to do so than others, and they have 
had their day. At present, what we fear the most in water are 
the germs of disease that it may contain, germs so exceedingly 
small that they pass through all usual filters. All at once, all 
these filters lost their popularity, and it has been necessary to 
find new types capable of attaining the object desired. 



THE BACTERIOLOGICAL WORLD. 93 

I am not here to describe them ; they are known enough. 
It is sufficient to say that they have solved the problem, and 
that water which has passed through a good ( improved, 
modern )* filter is absolutely free of microbes, whether it is acid 
or alkaline,pure or charged with organic matter. Unfortunately, 
to obtain these results, very fine porous porcelains, through 
which filtration is slow, are necessary. We may, in augmenting 
progressively the filtering surfaces, supply with sterilized water 
a home, a school, barracks, perhaps also a city, though this last 
result has not yet been realized ; but we can think to render 
thus harmless only drinking water. As to the water of general 
service it is, with the wants of modern hygiene, of too great a 
volume to be sterilized by these methods. 

Still it is important to render them as harmless as possible, 
for two principal reasons. The first is, that, if it contains per- 
nicious germs, some of these may be left on utensils or the clothes 
washed, on the floors,, and in the streets sprinkled. The second 
is, that, in the cities where there is not absolutely separate 
ducts for drinking water and washing water,, when, in case of 
scarcity of the former, may rise the temptation to replace it 
temporarily by the second, this temptation becomes irresistible 
if there is, to realize it, only to turn the key or manipulate a 
faucet. And it suffices that impure water penetrate but 
a moment in the pure-water ducts or pipes to deposit therein 
germs, on the ultimate evolution of which we know nothing, if 
not that, a priori, they have a chance in two to become 
dreadful. 

The problem of water for alimentation has been solved in 
various ways. Certain cities, as Paris, which have succeeded 
in procuring drinking water of excellent quality, depend on 
this, and do not think of their other waters, which they receive 
directly from a river, and which they send, without previous 
filtration, in the pipes, and sometimes in some pipes which 
should never receive anything but potable water. Other cities, 
less privileged, from a point of view of drinkable water, filter 
indistinctively all their waters, either by building laterally to 
their rivers certain forms of filtration dams, as Lyons and 



:: The editor. 



94 THE BACTERIOLOGICAL WORLD. 

Toulouse, or in making the water pass on artificial filters, as 
Berlin and Zurich. The two extremes of these solutions exist 
for Paris, on one side, in maintaining, during eleven months of 
the year, security against the diseases that water may conve}^ 
and promenading, during the last months of the year, the 
sprinkler of epidemic diseases in various quarters of the city, 
in order to create no jealousy ; on the other hand, Berlin, for 
example, in supplying, the whole year, to her inhabitants, 
mediumly good but not warranted water, but which they 
endeavor to render as pure as possible. It would be very 
difficult to know, and, so far as that goes, useless to seek, 
which of these two systems is the best. They are both 
bad ; they must be regarded as makeshifts, and we must seek 
to improve them. From this point of view, there has been 
made at Berlin and Zurich, on the use of industrial filters, 
curious observations which renew all that we thought we did 
know on filtration, and the interest of which is too great not to 
be given space in these Annates. 

The great industrial filters are, as we know, sand filters 
formed of a lower layer of washed pebbles, then a layer of 
gravel, then superposed layers of sand, finer and finer. To wet 
them regularly, the water is allowed to penetrate it slowly from 
the lower layer. When, in rising, the water has pushed before 
itself the air contained in the interstices of the sand, it is 
allowed to penetrate the filter from the top layer, and it filters 
under pressure from top to bottom. 

After numerous tests, the superiority of sand as filtering 
matter has been recognized, and when possible diluvial sand, 
principally the quartzy kind, containing the least possible pro- 
portion of carbonate of lime. This sand is in irregular grains, 
more or less fine, and, in noting that it filters properly only when 
it has attained a certain degree of tenuity, you might think 
that it is closely related to the constitution of porous porcelain, 
and that its mode of action is probably the same. If it were 
thus, its history need not be written, but, in reflecting a little, 
we see that it cannot be. 

In the first place, from a point of view of the attraction 
that the walls of a filter exercise on the minute solid bodies in 
suspension in the water, we know, by the tests of Mr. Kruger, 



THE BACTERIOLOGICAL WORLD. 95 

that sand is not worth the raw or cooked clay. Yet experience 
has taught the preference of the sand filters. It is true that 
they are filters of fine sand ; but, as the sand augments in fine- 
ness, the total volume of the empty spaces, left free between the 
grains, does not diminish as much as we might think. If the 
sand was entirely formed of grains of equal size and perfectly 
spherical, we could fill completely the space that it occupies in 
supposing each grain replaced by a cube, the side of which 
would be equal to its diameter, and the proportion of the empty 
spaces to the total volume would be equal to the proportion of 
the sum of the volumes of the cubes which would not fill the 
sphere which they contain. Now, a sphere placed in a cube, the 
height of which is equal to its diameter, fills, as we know, about 
half of the volume ; the proportion from the hollow to the full 
is then the unit, and the proportion from the hollow to the 
total volume is of about one-half. The size of the spherical 
grains may, therefore, decrease indefinitely ; if they remain 
equal, there will be always, theoretically, 500 litres of space per 
cubic metre. In practice, the existence of smaller grains, 
which lodge between the large ones, diminishes this volume a 
little, but it is curious to see that the proportion of the empty 
space to the total volume remains about constant as is sustained 
by the theory. In five kinds of finer and finer sand, studied 
from this point of view by Mr. Piefke, it has varied only from 
29 to 34 per cent., and, in a general manner we may admit, 
that, in the sandy, filtering mass, there is about one-third of 
space occupied by the air when it is dry, by the water when it 
is in operation as filter. Of what use, then, is the sand if it 
does not diminish the total volume of the spaces ? It augments 
the number of lacunary spaces, and consequently their total 
surface, and the mean proximity of the walls which limit them ; 
thence arise two advantages : one of a purely physical order, the 
other of intermediary order in physics and chemistry. 

The first advantage is to render uniform and regular the 
liquid current which passes through the filter. The moving 
power, represented. by the pressure of water weighing on the top 
of the filter, meets a resistance, which, as experience has 
proven, is in proportion at the same time to the thickness of 
the filter if the filter is homogeneous, and to the rapidity of the 



96 THE BACTERIOLOGICAL WOULD. 

liquid in the lacunary spaces, every time that this rapidity is 
not too great. When these lacunary spaces are irregular, water 
does not run through at a constant rate. But the law remains 
true for the mean rate. This is what Darcy had perceived, 
without demonstrating it with precision, I have made this 
demonstration for the porous walls, and Mr. Brunhes for the 
filtering masses of gravel. We can, therefore, say, basing our- 
selves on the experience, the equation 

v e = m h 
where li represents the pressure of water on the upper part of 
the filter, v the mean speed in the filter, and e its thickness. 
We express in this way that there is equality, that this equilib- 
rium between the power and the resistance, and that the 
movement of water is uniform with the speed v the value of 

which is 

h 
v = m — 
e 

To have an idea of what is the factor m introduced in the 
equality, we must suppose h = 1, and d = 1; that is to say, to 
represent to one's mind a filter of equal thickness for instance 
to 1 metre, operating under the pressure of 1 metre of water : 
we would have then v = m, which is the same as saying that 
m is, in metres, the swiftness of the current of water passing 
through the filter. This swiftness being evidently the slower 
in proportion as the filter is made up of finer sand, m dimin- 
ishes with the coarseness of the elements of the filter, and even 
much faster than they. It is impossible to give the law of 
variation when the lacunary spaces are irregular, but m varies 
as the fourth power of the diameter in capillary tubes. The 
diminution is, therefore, rapid when the diameter diminishes. 

Fine sand insures us, beneath an average thickness, the slow 
circulation of the liquid which passes through it, and by that 
the regularity of the action of the lacunary walls on the water of 
the filter. This action, on which we have often insisted in these 
Annates is a molecular action being exercised at a distance on 
the solid b6iies in suspension, and immobilizing them in con- 
tact with the wall by the same mechanism that fixes a coloring 
matter to the surface or in the structure of the fibres of a tissue. 
It is here that we find the inferiority, already mentioned from 



THE BACTERIOLOGICAL WORLD. 97 

is point of view, of a wall of silicious sand on the wail of 

iy, and as, besides, our lacunary spaces are certainly very 

egular we have the right to doubt, a priori, that this filter 

erates as a filter of porcelain, and arrests all the microbes in 

spension in the water which runs through it. 

To know this, we have only to repeat an interesting experi- 

mt of Mr. Piefke, who after having filled with sterilized 

id a filter, the walls of which had been washed with a solu- 

>n of corrosive sublimate, and after having besides taken the 

ecaution to leave the whole during twenty-four hours in con- 

3t with a solution of sublimate, evacuated the antiseptic 

uid, and caused his filter to' operate in the ordinary fashion, 

counting the number of colonies ( of microbes ) as furnished 

each cubic centimetre of water before and after filtration. 

Instead of diminution in the number of microbes, it is an 

gmentation that he has noticed, at least in the first days. 

Dreover, not only was the water sterilized in this contrary 

mion (that is, not sterilized at all) but, from the point of 

3W of limpidity, the filtration was most imperfect. Little by 

tie the filter became obstructed. The exit of water under a 

ren load diminished, and it was necessary to augment the 

essnre to obtain this exit at the same level. Little by little, 

so, there was formed on the superior surface of sand grayish 

icous layers, formed of a net work of filaments of algas, 

crobes, diatomes, the whole imbedding the sedimentary, 

ineral and organic matters that any water usually contains. 

5 this surface living layer became thicker the filter operated 

tter, the water was more limpid and contained less microbes, 

it it is scarcely before the period of two months that this filter 

ve acceptable results, that is, that the number of bacteria in 

e filtered water was reduced to a few dozen per cubic centi- 

3tre. 

The ordinary filters, made with diluvian sand, simply 

ished and populated of microbes, arrived more rapidly at the 

rviceable state which is characterized by the term ripe. - But 

ey passed through the same phases : at the origin, they filtered 

dly, and it is only when the surface was covered by this 

uddy coating which we have just mentioned, that the number 

bacteria in water filtering through it is reduced to its 

inimum. 



98 THE BACTERIOLOGICAL WORLD. 

We arrive, therefore, at this apparently paradoxical con- 
clusion, that it is^the bacterial surface coating of the filter, and 
not the sand which fills it, which retains the bacteria of water. 
In reflecting on this the subject is not so astonishing. When 
we filter an acid precipitate of sulphate of baryta on filter 
paper, the liquor that passes through is at first cloudy, and it 
is only after this sulphate precipitate has itself lined the bot- 
tom of the filter that the same sulphate is completely retained, 
and that the liquid is clear. This coating of entangled bacterial 
filaments is thus much less permeable than sand. What proves 
it is that it is necessary to augment the pressure in proportion 
that this coating thickens, in order to keepthe filter in a con- 
dition to render its ordinary supply, and it is doubtless this 
coating which opposes itself to the passage and reduces the 
value of the coefficient m in the formula above given ; for, when 
we lift it up, the sandy layers regain at once their old per- 
meability. We understand very well, therefore, that this upper 
coating ends in forming a filtering wall as soon as it has cov- 
ered the surface ; it is a living filter, instead of being a mineral 
one. 

But this conclusion, in its turn, raises a number of ques- 
tions to which we have yet no answer. If the sand serves only 
as support to the true filter, why has experience taught us to 
give it this thickness, which is, at the minimum, with the fine 
sand, of 60 centimetres. This thickness of sand is traversed 
by the microbes, for they are always to be found in the dripping 
filtered water. They may even swarm there and multiply, as 
demonstrated in the experiments of Mr. Piefke, which we cited 
a while ago. How is it that they do not fill the filter com- 
pletely ? Finally, whence comes this continual exit of bacteria 
with the water that comes from the filter ? Does it come from 
the surface, or from the depth of the filter ? To solve these 
questions we must study the composition of a ripe filter at the 
moment of its most perfect operation. In the beginning, this 
filter was formed of homogeneous sand, not washed ; the 
microbes were equally distributed throughout its whole thick- 
ness. When it is ripe a new study shows that the microbes 
have augmented in number, but unequally in various layers. 
At the bottom of the filter the multiplication has been poor, 



THE BACTERIOLOGICAL WORLD. 99 

but the populating is more and more abundant as we ascend 
towards the surface, and, on the surface itself, the number of 
germs per cubic centimetre is very great in comparison to what 
we find in a cubic centimetre of sand from the lower layer. 

We understand without trouble that experience should 
have taught to modify the speed of the current of water 
through a filter thus constituted. The sand, a nonporous 
matter, is not penetrated by the microbes, and serves as support 
for them ; but a current a little rapid would sweep them away, 
and does in fact sweep them away. We must, therefore, cause 
the filter to operate under light pressure, and not exact from it 
a quantity of water beyond a certain supply. The filters of 
Berlin scarcely surpass the limit of 100 millimetres per hour. 
A more rapid current would force the impurities that cover 
the surface to penetrate deeper in the thickness of the filter, 
would augment by that means the resistance and reduce the 
supply. On the other hand, it would carry in larger proportion 
the microbes and nutritive organic matters that water contains. 

It is true that these organic matters are in small quantity, 
and at first there seems to be an enormous disproportion 
between the weight of the alimentary matter and the number 
of beings struggling for it. A litre of water from the Spree, 
tor instance, contains about 80 milligrams of organic matter, 
in counting as such, all that which m the evaporative residue 
of one litre disappears when calcinated at a mild heat, and 
without decomposing the carbonates. On the other hand, we 
find often in it 100,000 germs per cubic centimetre, that is, 
100,000,000 per litre. But these 100,000,000 of germs, in 
supposing each to be 1-1000 of a millimetre, and attributing 
to them the density of water, do not weigh 1-10 of a milligram, 
which comes to saying that the weight of the nutritive matter 
is about 800 times the weight of the living beings that may 
consume it. 

This nutritive matter, therefore, is not wanting in 
quantity, but perhaps we would be less satisfied if we analyzed 
it as to quality. From this point of view we have only 
incomplete information on the subject, due to the studies that 
we will resume further. But if we ask ourselves, a priori, 
whence come particularly the organic matters that we may find 



100 THE BACTERIOLOGICAL WORLD. 

in water, the general notions that we have on the organic 
economy of the world will answer that they are formed chiefly 
of the product of microbic life. 

Vegetables and animals have for property to render 
insoluble in water the substances of which they nourish them- 
selves, and all that which they reject that is soluble in water, 
for instance in the urine of animals, is formed either of inuti- 
lizable products or products used by the cellular life. It is 
after they are dead that microbes render soluble in water, 
either by their diastases, or their vital action, the materials of 
superior beings. Hardly are they dissolved, that other microbes 
seize upon them. And, in a word, the organic materials in 
solution in water must be formed, if not totally, at least in a 
large proportion, of substances having undergone in a more or 
less pronounced manner the action of cellular life or that of 
microbic life, and have become by that means less proper to 
nourish new beings. We will soon have occasion to give import- 
ance to this conclusion. 

It would be very useful to report this conclusion directly 
by experiment. Unfortunately organic matter of water has 
never been studied except by so-called practical means which, 
invented in England for a pressing need, have become implanted 
we know not why in science, and enjoy a reputation that we 
cannot explain. I mean the quantitative analysis of the 
organic matter by the hypermanganate potash in acid or 
alkaline solution, and in general all those processes in which, 
instead of determining by weight the total quantity of organic 
matter contained in a given water, it is attempted to analyze it 
indirectly, such as for instance, by the quantity of oxygen which 
it needs to burn. 

When organic matter is heterogeneous, as necessarily is that 
which is in solution in the water, we cannot establish its weight 
from the oxygen necessary to burn it ; for its different elements 
need for complete combustion different proportions of oxygen. 
The sugar, for instance, needs about its weight, the alcohol the 
double of its weight. We could therefore conclude nothing 
from the total quantity of oxygen liberated by a given weight 
of hypermanganate of potash, even though this hyperman- 
ganate of potash would push to the very bottom the oxidation 



THE BACTERIOLOGICAL WORLD. 101 

of organic matter on which it is made to act. As to the 
already old experiments of Messrs. Tiemann and Preusse, 
there is not a substance, except oxalic acid, that can borrow 
from hypermanganate of potash all the oxygen it needs to 
burn, and they would even borrow from it very different 
fractions : Tartaric acid, 4-5 ; sugarcane and glucose, 1-2 ; 
tyrosine, 1- 3 ; asparagine, 1-9 ; leucine, 1-10 ; benzoic acid and 
allantoine, 1-40 ; urea, none at all. We may add to this cause 
of uncertainty, resulting from the fact that the weight of 
organic matter is not proportional to the weight of the oxygen 
necessary to burn it, another resulting from the fact that 
hypermanganate of potash in acid or alkaline solution con. 
sumes the various organic matters very unequally. It is as if 
to find the weight of a body whatsoever, one would weigh an 
unknown variable fraction of it in a scale, one of the dishes of 
which would be moving blindly on the arm of the corresponding 
lever. There is, therefore, nothing precise to conclude from 
analysis of water by hypermanganate, whatever be the method 
adopted. It is to put in the hands of investigators not only a 
false but capricious scale, with the only argument that the 
weighing is easy. 

All that can be said in favor of this method is that, when it 
is applied to the same water before and after filtration, it may 
give a good idea of the loss of organic matter during filtration. 
We may add, but always with a great deal of reserve, that- the 
organic matter the most affected by the hypermanganate and 
acid solution being, after Messrs. Tiemann and Preusse, the 
most complex matters, the most nutritive for microbes, the 
most remote from the state to which microbic life brings them ; 
it will be in preference to these matters that one must attribute 
the differences in analysis by hypermanganate before and after 
filtration. 

We know how this analysis is made, by the aid of the dis- 
solution containing 0gr,32 of hypermanganate of potash .per 
litre and treated by means of a solution of oxalic acid. For 
instance 300cc. of water with an addition of 5cc. of sulphuric 
acid, diluted with twice the volume of water, with 10 or 20cc. 
of the solution of hypermanganate. After five minutes of 
boiling, a fixed period adopted, the liquid is not yet decolorized. 



102 THE BACTERIOLOGICAL WORLD. 

We complete the decolorization in adding a known quantity of 
the oxalic acid solution, and we treat the oxalic acid remaining 
in adding again hypermanganate until the reappearance of a 
persistent red color. A simple calculation shows the amount 
of hypermanganate destroyed by the organic matter of the 
volume of water used. 

We designate generally in treatise on this matter, under 
the name of oxidability, the number of cubic centimetres of 
experiment liquor decolorized by 1 litre of water, after these 
five minutes of ebullition. In taking ten minutes, the quantity 
would be slightly increased. We may, instead of taking into 
account the number of cubic centimetres of the liquor of hyper- 
manganate, calculate the weight of hypermanganate that was 
in it, or again calculate the oxygen that it may have yielded. 
With the method adopted lcc.=0gr,.0008 of oxygen. Each 
savant has on this his habits in which it is useless to disturb 
him, for his results being always comparative, it matters not to 
what unit his figures relate. It is only necessary that he tells 
it so that, if need be, one may make the translations. 

From what we have said above, this comparison does not 
inform us in the comparison of the materials of the water, the 
presence of which we would have the greatest interest to reveal. 
It cannot be either a matter of indifference to us to know if 
water has received oozings from cess pools, and the urea, which 
might warn us of the danger, the benzoic acid of the urine of 
herbivorous animals, and the amide products, as leucine, 
tyrosine, glycocol, produced by the transformation of nitro- 
genized matter are little sensitive to the action of hyper- 
manganate of potash in acid solution, perhaps for the same 
reason which renders them very resistent in face of the 
oxidizing combustion of the organism. — Dx. Annates de 
L'Institut Pasteur. 

[ To he continued.'] 



THE BACTERIOLOGICAL WORLD. 103 

MEDICAL BACTERIOLOGY. 



METHYL VIOLET. 

( Or the fancy copyrighted name Pyoktanin.) 

BY FLAVEL TIFFANY, M. D., 

Kansas City, Mo. 

A few months agoDr, J. Stilling, of Strasbourg, gave to the 
world in the April number of Revue Generate D' Opthalmologie 
his experience in the use of methyl violet in the treatment of 
various affections, more especially those of the eye. 

When the article first appeared, May, 1890, I took the 
liberty of translating it, and, subsequently, my translation 
was published in two different numbers ( not consecutive) of 
Dr. Lamphear's Kansas City Medical Journal. I commenced 
the use of methyl violet from the day I first read Prof. Sell- 
ing's article, for, at that time, I had on hand a case of suppura- 
tive iritis which had resisted all treatment, continuing step by 
step through the several stages of inflammation, until panoph- 
thalmitis seemed inevitable. The patient was an. old man of 
seventy -two years ; of rather feeble physique, upon whom I had 
made extraction of a nuclear cataract some eight weeks 
previous to the attack of iritis. He had made a good recovery 
from the operation and with fair results, viz.: With -f- 11 
Dv. 20-40. Could read number 2 with + 14 D. There was 
a slight amount of cortical substance remaining. The iritis 
did not ensue for a month after he had gone from the 
hospital, when it appeared quite suddenly one day, as he 
was sitting in the harvest field watching the harvesters at 
work ; he says as he crossed the lot from one shade tree to 
another in the full blaze of the bright harvest sun, he felt a 
sharp pain dart through the eye, and from that time he was 
not wholly free from pain, which gradually increased with 
intolerance of light and diminution of vision. It was several 
days after the attack before the patient returned to the city. 
When he presented himself I found an intense inflammation of 
the iris with plastic exudation into the front chambers. 



3 04 THE BACTERIOLOGICAL WORLD. 

Mydriatics and hot water applications were scrupulously 
used in connection with other anti-phlogistics, but all to no 
purpose, the inflammation mounted higher and higher and the 
exudation was more profuse and of a suppurative character, the 
iris becoming a deeper red until finally the whole chamber became 
blood-red ; the cornea began to grow steamy ; there was intense 
ciliary injection ; in fact the whole ball was aglow, and the lids 
swollen. 

There was much pain and total blindness, and I despaired 
of any chance of saving the ball, much less any sight. 

It was at this stage, that I read Dr. Stilling's article on 
methyl violet. I immediately had a preparation made, 1 to 1000, 
which I had dropped into the eye, one or two drops at a time, 
thrice a day. This caused no irritation, but for few hours there 
was little, if any, amelioration of the inflammation, although 
the eye felt better. At the end of the second day, there was 
perceptible abatement of the congestion, and the pain was 
nearly gone. 

The amount of the medicine was increased. The con- 
junctival sac was filled at each application, morning, noon and 
night, and the inflammation gradually subsided. 

The me.thyl violet was continued for about three weeks 
when the eyeball had become clear and free from inflamma- 
tion. The shape and size of the ball had been perfectly con- 
served, the iris had assumed its natural color and brilliancy, 
although the pupil was nearly occluded, and there was only per- 
ception of sight. The patient returned to his home, in Kansas, 
with the hope, through the means of a subsequent operation, 
to be made in a few months, of regaining useful vision.* 

It has been about six or seven months since I commenced 
the use of methyl violet in affections of the eye. 

I have used it a great many times, and in many patients, 
and have kept a careful record of several of the cases. It has 
been mostly used in phlyctenular conjunctivitis and granular 
conjunctivitis. 

In cases of trachoma, when treatment cannot be continued 
by me on account of patients being obliged to return to their 
homes out of the city, it has become my custom to prescribe a 

*Since heard that he could see to count fingers. 



THE BACTERIOLOGICAL WORLD. 105 

bottle of methyl violet ( 1 to 1000 ) to use at home three times 
a day, and it has been invariably the case, that these patients 
go on to recovery without any relapses, and that the medicine 
never acts as an irritant, but is always soothing and agreeable. 
I have used it with excellent effect in dacryocystitis by means 
of the lachrymal syringe. I have also used it in otitis media 
purulenta, but not with as marked benefit as in affections of 
the eye and lachrymal apparatus, and nasal catarrh. 

I have employed this agent in microscopy as a staining 
material, knowing its special and thorough action as a stain 
for different forms of microbes, and especially for the micro- 
cocci, and that it always means death to the bacillus when it comes 
in contact with it, striking, as it were, to the heart or nucleus 
of all cells, paralyzing all vital action at once ; but it had not 
occurred to me to employ it as a therapeutic agent even in those 
diseases which are generally conceded to have their etiology in 
the microbes, until my attention was called to it by Prof. 
Stilling's able article, to which I have referred. 

Since I have been using this agent I have had the oppor- 
tunity of employing it in a single case of gonorrheal ophthalmia. 

As yet, of course, the remedy has not been sufficiently 
used to establish it as so valuable a remedy as Dr. Stilling 
would claim for it. 

Methyl violet may be used both as a local topical, and 
general system agent, in fluid, or pomade, or spray one- 
twentieth to one per cent. It remains to be seen if it shall 
prove a valuable remedy in pulmonary affections, in pyemia, etc. 

If it has a special predilection for the bacilli and cocci, 
we shall look for it as a boon in most diseases. 

Within the last few days I have had occasion to use the 
methyl violet in two cases : One of irido-cyclitis and the 
other choroido-cycloiritis, both of a most serious nature. That 
of iridocyclitis occurred in a reverend gentleman, Mr, D., 
aged fifty-five ; blue eyes. This patient consulted me December 
9, complaining of quite a severe pain in the right eye with 
dimness of vision. Examination revealed acute inflammation of 
the iris with a more intense deep-red congestion of the sub- 
conjunctival vessels over the entire anterior scleral surface. 
The pupil was contracted and inactive, and would not respond 



106 THE BACTERIOLOGICAL WORLD. 

in the least to mydriatics. A one-per-cent. solution of sulphate 
of atropia, dropped into the eye five or six times a day, made 
no impression whatever, not even controlling in the least the 
photophobia or congestion ; in fact all the symptoms grew 
steadily worse, and on the third day the cornea lost its trans- 
parency, became steamy, so that the iris was nearly hidden and 
the vision was gone. Could not see to count fingers ; the 
fellow's eye had taken on symptoms of irritation, and its vision 
was considerably reduced. At this period December 11, I 
ordered methyl violet one-seventieth per cent, dropped into the 
eye every hour in connection with the sulphate of atropia, and 
to my great delight the next day, December 12, I found the 
pupil dilated to more than twice its size of the day before, the 
congestion subsiding, the cornea clearer, and the vision sufficient 
to count fingers. The treatment was continued, and on the 
following day the patient was able to recognize a person across 
the room, pain was gone, redness less and pupil widely dilated ; 
December 15, pupil wide open and vision 20-80, eye fast 
recovering. The methyl violet has quite a mydriatic effect, 
and also controls the power of accommodation to a considerable 
extent, though it is claimed by some that it does not affect the 
power of accommodation. This fact I have verified many 
times by using it alone without atropia. In the above case I 
feel confident that the methyl violet, from its diffusability and 
germicidic effect on the pyogenic microbe, cut short the disease 
and so saved the eye. I do not believe that in this case the 
mydriatic effect of the atropia could have obtained without the 
methyl violet, besides the methyl violet controlled the irritating 
effect of the atropia to the cornea and checked the keratitis. 

The other case, choroido-cycloiritis, occurred in a young 
man of twenty-five years ; dark brown eyes ; A. B. Patterson, 
from Nebraska, a station agent on the Missouri Pacific Railroad. 

The history as gained from the patient was as follows : 
" While I was walking down the street, October 13, with a pen- 
knife open in my hand, I met a friend who, in play, knocked 
my hand with the knife so that the open blade pierced my right 
cheek, going into the eye, and then the doctor put a stitch in 
the eyeball, and assured me that the sight would be all right 
(a German doctor). For about ten days the sight was very 



THE BACTERIOLOGICAL WORLD. 107 

good, and there was but little pain, and then the eye began to 

pain and I could not bear the light and the sight gradually 

diminished and the pain grew worse/" 

An examination revealed a scar of the right cheek, lower lid 

and. eyeball. That of the eyeball about 10 mm. long, about 

one-third, of which extended, into the cornea from the inner 

inferior region, near the attachment of the inferior rectus 

muscle. 

The cut of the cornea embraced the entire layers of the 

cornea, and evidently the blade went through the sclera and 

choroidal, though there was no evidence of its cutting the iris. 

The pupil was contracted and the vision was not sufficient to 

count fingers ; photophobia intense. I ordered one-per-cent. 

solution of atropia to be dropped into the eye every two hours, 

night and day. The second day the patient was sent to my 

rooms, the mydriatic had not had the least effect. The 

ophthalmoscope revealed a grayish light-colored body, with 

small pale blood vessels mounting up over it, protecting from 

the region of the ciliary body, and the sclera immediately 

exterior was swollen or bulged. There was evidently detached 

retina, with exudation beneath it. At this date I commenced 

the use of methyl violet, and had it dropped into the eye every 

hour night and day. On the following day there was some 

dilation of the pupil and less pain and soreness. December 16, 

I injected a one-tenth-per-cent. solution 10 m. in the eye 

through the sclera. The soreness subsided ; the pupil became 

more open although the exudation was much the same. Dr. 

Stilling speaks of injecting the vitreous of a rabbit without 

harm to the eye. 

My experience so far has been that the good effect from 
this agent is especially to be realized in inflammation of the 
deeper structures of the eyeball, owing, probably, to the great 
diffusability and penetrability of the aniline traversing almost 
immediately the cornea, and sclera to the chambers of the 
eyeball, and having for its predilection the bacillus and micro- 
cocci. In ulceration of the cornea, I have used it with most 
excellent results in the form of pomade 1 to 200. 

For marginal blephoritis the methyl violet pomade, one- 
half per cent., carefully worked into the roots of the lashes, by 
means of a spatula, works a speedy cure. 



108 THE BACTERIOLOGICAL WORLD. 

Prof. Stilling, from his bacteriological experiment, found 
that milk mixed with methyl violet would not sour, nor would 
butter become rancid ; that urine even could remain in a 
thermostat at 32° C. for a week without putrefaction or pre- 
senting any bacteria whatever ; in fact, any substance containing 
a solution of methyl violet, of even 1 to 32,000, is absolutely 
indemnified. 

This agent acts as an antiseptic, killing the pyogenic 
bacteria, and, from its d iff: us ability and non-destructiveness to 
tissues, it is superior to other known antiseptics, and especially 
to thermocautery which is so efficient a germicide, but can only 
be used at limited points. ( Eead before the Missouri Valley 
Medical Association, Kansas City, December, 1890.) 



CONTRIBUTION TO THE STUDY OF ETIOLOGY AND PATHO- 
LOGICAL ANATOMY OF THE ABSCESSES OF THE LIVER. 

RESEARCHES BY PROF. AL. DEL RIO.* 

Dr. Alejandro del Rio, assistant professor of pathological 
anatomy, at the University of Santiago, published in the 
Revista Medica of Chili an important work on the abscesses of 
the liver (suppurative hepatitis), a malady of the warm 
countries., which represents from 5 to 100 of the mortality' in 
the hospitals of men at Santiago. 

" If the actual tendence of the medicine," says Mr. Del Rio, 
i ' is to employ the etiology for the classification of diseases, we 
should not forget that this knowledge has not reached every- 
where the extent and solidity necessary to overturn the patho- 
logical anatomy from the place which it occupied." 

" We cannot deny that the knowledge of these causes has 
produced in medicine a deep and beneficial revolution, and that 
the benefit is so clear and evident that it cannot be denied by 
skeptics. But to take bacteriology for a unique guide, in 
which the light is subject to many illusions, is still premature." 



*Revista Medica of Chili, November, 1889, in 8 with 5 plates, work 
crowned by the Medical Congress of Chili in 1889 ( text in Spanish ) . 



THE BACTERIOLOGICAL WORLD. 109 

" More practical and more positive results are obtained by 
the aid of these two sciences. Such is the rule that we follow 
in this study." 

This is in fact the plan that Mr. Del Eio follows in the 
work which has for an object to seek the causes of tropical 
hepatitis, to study its nature rather than make a clinical and 
therapeutical study. 

The author thinks that this malady is due to some external 
causes, and that its development is secondary, consecutive to 
other processes which have accomplished their evolution ; that 
it is an infectious disease producing some anatomical lesions 
similar to those of other maladies of same nature, not due to 
common causes as cold, heat, traumatism, but to some special, 
external, organized agents. 

He commences then by giving an extremely complete and 
careful description of the anatomical lesions, then a histolog- 
ical study of the affected tissues and of the morbid products, 
finally the summing up of the bacteriological researches 
executed with the aid of the best methods and of the most per- 
fect instruments. Mr. Del Rio has used in fact, some apochro- 
matic objectives of Zeiss, accompanied by their compensating 
oculars. 

He found thus, both in the pus of the hepatitic abscesses 
and in the disorganized tissues from their walls, various micro- 
organisms, micrococci united, two by two, and forming groups, 
more or less large ; in another case, larger micrococci ; in a 
third, some bacilli of small dimensions confined to the middle, 
and the poles of which were less colored than the middle por- 
tion ( gentian violet ) ; in a fourth, bacilli taking less color at 
the poles, and presenting at the center a clear spot. 

These details are represented in the colored figures, 
extremely pretty and of an excessively fine design. 

Mr. Del Rio is led to think that these micro-organisms pene- 
trate through the intestine which, 87 times in 100, presents 
dysenteric ulcerations ; taken by the ramifications of the vena 
porta, they reach the liver of which the)" determine the decom- 
position. In fact, abscesses of the liver present clearly a 
necrobiotic character rather than phlegmasic. 



110 THE BACTERIOLOGICAL WORLD. 

" We find in our books of medicine that the intestinal 
ulcerations are often a complication of the hepatitis of the warm 
countries. We see the abuse which is made daily of the word 
" complication." The intestinal ulcerations are not complica- 
tions of the hepatitis : they do not accompany it ; they are very 
probably the place of entrance of the disease. On the other 
hand, should the lesions not exist, the epithelium of the intes- 
tine does not oppose an impassable barrier to the parasitic 
micro-organisms, especially in a country like Chili, where intes- 
tinal chronic catarrh is very frequent, as well as other lighter 
affections of the intestines which is accompanied with an active 
epithelial desquamation." 

" But it is the dysentery which appears to furnish more 
often the place of entrance. The two maladies coincide in the 
proportion of 87.37 to 100." 

From a study of the geographical distribution of the sup- 
purative hepatitis in Europe, Asia, Oceanica, Africa and the 
two Americas, we find that this malady is more frequent in the 
countries where the intestinal affections are more common. 

" In Chili, in particular, where the abscess of the liver is 
frequent, 11 per cent, of the general mortality in hospitals is 
due to dysentery." 

" The predisposing or general causes are the climate, the 
variable temperature, with the latitude and altitude of the 
different regions of the Chilian belt, the humidity of the air, a 
bad or exaggerated alimentation and alcoholic drinks. The 
alcoholism has taken, in Chili, on account of its frequency, an 
exceptional gravity," 

i( At the rate we go, said Mr. Del Kio, our race, proverb- 
ially strong and robust, skilful in all kinds of work, especially 
those requiring strength and resistance to the rigors of the 
climate, is marching with a rapid step towards the most com- 
plete ruin. In fact, it is sad to confess it, it is rare to find in 
our people an individual who is not alcoholic in the full sense 
of the word. All, and particularly those of the class of workers, 
the peon, on account of the particular alcohol (it is the alcohol 
of the worst kind of grain ) which they absorb daily, pass 
several days of the week in complete -intoxication, oftener 
exposed to all the inclemency of the weather." 



THE BACTERIOLOGICAL WORLD. Ill 

<c Do not think," added he, " that I exaggerate. When we 
demand of the individual affected with hepatic abscesses as to 
what cause they attribute their malady, two-thirds attribute it 
to great carousals or to remaining without shelter, exposed to 
the weather, while they were intoxicated. " 

A remarkable fact, and which proves well the role that 
intestinal affections play in the production of the abscess of the 
liver, is a parallel march of dysentery and of hepatitis. Mr. 
Del Eio represents by the aid of graphics the results furnished 
by the statistics taken since 1870, and we find the maxima or 
minima in the localities and in the years where occurred the 
maxima and minima of dysentery. 

Moreover, as the hygienic conditions in which the popula- 
tions live improve, and they improve year by year, thanks to 
the efforts of the administration, the diseases of the intestine 
diminish, and at the same time the abscesses of the liver. 

So that, were it not for alcoholism, this cause of disease 
which does not diminish, dysentery as well as hepatitis are two 
affections which, declining in frequency every year, would dis- 
appear some day. 

"We cannot insist further on the work of Prof. A. del Rio. 
This is not only the best that we know on this subject, but we 
give it as a model both from the standpoint of method and con- 
cerning the anatomical and bacteriological researches. 

This work of Mr. Del Rio obtained last year the prize 
given by the Medical Congress of Chili. — Journal de Micro- 
graphie. 



112 THE BACTERIOLOGICAL WORLD. 

HYGIENIC BACTERIOLOGY. 



REVIEW OF M. ZAGARPS RESEARCHES ON THE MECHAN- 
ISM OF THE ATTENUATION OF RABIC VIRUS. 

In a work which has been analyzed in these Annates, Mr. 
Protopopoff has drawn the conclusion that it is chiefly the heat, 
and not the desiccation, which intervenes to attenuate the virus 
of rabic marrow, which is used in vaccination by the Pasteur 
method. I had made the observation on this subject that 
we remained in the vague so long as we are contented to say it 
is the beat which acts, and that it would be useful to search 
how it acts 3 and if it is not in putting in play a phenomenon of 
oxidation. 

This call to new researches has been answered by Mr. 
Zagari, who has studied comparatively the influence of the 
sojourn of the marrow in the air, in the pneumatic space, in 
carbonic acid and in oxygen. In each of these series of 
experiments he has varied the conditions of preservation, in 
leaving, or not, potash in the presence of spinal marrow, sus- 
pended in a drying flask, or immerged in sterilized, distilled 
water, or a broth mixed in glycerine in equal parts. 

Lastly, he has made some comparative experiments at 20° 
C. and 35° 0., in order to see the influence of the heat. This 
last is not doubtful. Whatever be the mode of preservation, 
the virulence lasts longer at 20° than at 35°. For example, 
through the air, and in the presence of potash, that is to say, 
in the ordinary conditions of preservation of the marrow in the 
various anti-rabic institutes, the virulence is not extinguished at 
the end of 10 days passed at 20°, whilst it disappears at the end 
of 66 hours at 35°. 

This being well established, it was natural to operate on 
this last temperature in order to study the mechanism of the 
action of the heat. This action is then more rapid, and 
the experiments shorter. Without my insisting, we guess the 
disposition of these experiments. I will say, only, that, in 
order to make the vacuum on the marrow, Mr. Zagari recom- 
mends highly the use of mercurial pumps, which we are 
astonished, in fact, to see so much neglected, and replaced by 
complicated arrangements which are not worth them. 



THE BACTERIOLOGICAL WORLD. 113 

The heat has, for sure effect, to accelerate the desiccation of 
the marrow. Let us see, in the first place, if the desiccation 
does not intervene by itself, contrary to the opinion of Mr. 
Protopopoff. it suffices to compare between them the marrows 
preserved in the presence of the potash or those im merged in 
water or the glycerinated broth. We find thus that the virulence 
remains nearly twice as long in the immerged marrow; thus 
the desiccation has a special action, independent of that of the 
heat. But the heat, which certainly produces a desiccation 
more rapid, is able, also, eventually, to hasten the phenomena 
of oxidation. In order to know what their proper action is, it 
suffices to compare the marrows, preserved in the same conditions 
in the air, in the vacuum, and in the presence of carbonic acid. 
Mr. Zagari has thus found, among other results : 

First. That the marrows suspended in a drying flask, in 
the absence of potassium, lose their virulence after 56 hours in 
the air, but still retain it after 121 hours in the vacuum, and 
after 198 hours in carbonic acid. 

Second. That the marrows suspended in the presence of 
potassium lose their virulence after 66 hours passed in the air, 
and retain it after 116 hours in the vacuum. 

These results, to which we could add those drawn from the 
comparison of marrows preserved in broth or in water, suffice 
to show that the heat puts in action a phenomenon of oxidation. 

Does this action reduce itself to this phenomenon of 
oxidation ? One would think not, in noting that the marrow 
attenuates itself in the hands of Mr. Zagari, notwithstanding 
the minute precautions taken by this savant in order to free it 
from oxygen. But it would be concluding too hastily, and Mr. 
Zagari leaves, with wisdom, the question in suspense. The 
marrow is in contact with the air while we extract it from the 
medullary canal, the oxygen contracts with it some of those 
unstable combinations, analogous to that which it contracts 
with the haemoglobin, that the pump of mercury does not 
destroy, and which can serve as ulterior oxidations. It is true 
that this source of oxygen is of the most mediocre, but it does 
not take much gas to oxidize a matter so imponderable as a virus. 
The question remains, then, pending. It has, on the other 



114 THE BACTERIOLOGICAL WOELD. 

hand, moderate importance, and the main, essential point was 
to show, as Mr. Zagari did, the intervention of a phenomenon 
of oxidation in the attenuation of rabic virus. — Dx. in 
Annates de L' Institut Pasteur. 



BACTERIA AND PLANT DISEASES. 



BY T. J. BURRILL, PH. D. 

Bacteria were long known as having a causal relation to 
disease in animals, before they were in plants. In De Bary's 
Vorles ungen ueber Bacterien, published in 1886, he tells us 
that : " Parasitic bacteria do not often occur, according to 
our present experience, as the contagia of plants." Now 
De Bary was, at this time, the best botanist in Europe making 
a leading specialty of fungi, and in particular of parasitic fungi, 
hence of the diseases of plants. He also gave prominent atten- 
tion to bacteria, as his book just named gives ample evidence. 
As a matter of fact, the first paper, upon a disease of growing 
plants due to bacteria, was read in August, 1880, before the 
American Association for the Advancement of Science, by the 
present writer, upon the so-called blight of pear and apple 
trees. Since that time, there have been too many plant 
diseases attributed to these organisms to make De Bary's state- 
ment true for to-day. Several of these have been made known 
by European investigators, but, in this one particular, the 
Americans seem to have taken, and to have kept, the lead. 
More of such diseases have been worked out and described in 
our country than anywhere else, — not probably because we 
really have more of them, but because attention has been 
specially directed -towards them. The known number is also 
rapidly increasing, due to the activity of several investigators, 
and to the recognized importance of the studies. 

The literature upon the subject is, however, widely scat- 
tered. There exists no other publication in the world in 
which a department is provided for this special purpose. The 
existing periodicals of the Old World, devoted to bacteria and 
their allies, are concerned alone with the diseases of animals, 



THE BACTERIOLOGICAL WORLD. 115 

except for occasional references to those of plants. On the 
other side, as with us, the botanical journals have been the 
main avenues by which authors upon our special topic have 
sought the public eye. In the want of a more specialized pub- 
lication, this was the best that could be done, yet it has no 
doubt occurred to others that the diseases of animals and of 
plants, due to the same class of parasites, ought to find publica- 
tion together. This is true whether or not the matter is also 
contributed to the more general journals, in the future as in 
the past. The results of investigations in one kind of host 
will very probably aid the investigator upon the other hosts. 
There is also much probability that in some cases the self-same 
organism holds a causal relation to certain diseases of both 
plants and animals — a thing not heretofore sufficiently appre- 
ciated. The question of the origin of "disease germs " is still 
an exceedingly important one, and possible contributions to 
knowledge here may come from studies upon the bacterial dis- 
eases of plants. 

There is still another reason for which these parasitic 
diseases of plants should receive more attention on the part of 
bacteriologists. No living animal tissues can be sectioned in 
the microtome, and the thin slices studied under a high-power 
objective. But this can be done with plant tissues. There is 
far greater hope of being able to find out in the latter case just 
what action the invaders have in the regular course of their 
development, in what special manner they accomplish their 
destructive results, and just what favors or hinders their pro- 
gress. If auimal bodies could be thus taken apart and the 
small pieces still retain their vital activity, it is easy to suppose 
that many obscure problems would have been cleared up before 
this time, which now remain obscure. It will no doubt be an 
excellent thing for the animal pathologist or bacteriologist to 
acquaint himself thoroughly with the bacterial diseases of 
plants, and it certainly is desirable that he who would study 
such maladies of plants should keep abreast with what is 
accomplished in the studies upon the transmissible diseases of 
animals. 

It is proposed that some account shall be given in this 
department of the Bacteriological World of every disease 



116 THE BACTERIOLOGICAL WORLD. 

known in plants, both at home and abroad, which is attributed 
to bacteria, and at the same time the endeavor will be made to 
gather every item of information of possible bearing upon the 
study of these diseases. To this end investigators and authors 
are solicited to contribute results of research in formal papers, 
and to freely communicate items of all kinds and of every 
degree of importance bearing upon the subject. By all means 
let us have the little observations and discoveries too likely to 
be considered insignificant. Nothing of the kind can be too 
little. What have you now of this nature ? Please send it 
along.* 

* Address contributions on this subject to Prof. T. J. Burrill, 
Champaign, Illinois. — Ed.] 



ORIGINAL ARTICLES. 



THE PRINCIPLES OF ANT I- R ABIC VACCINATION —Continued. 



BY PAUL GIBIER, M. D., 

Director of the Pasteur Institute, New York. 
( Continued from January Number.) 

4th. After inoculation the animal survives; immunity 
has been acquired. Though it is somewhat inconstant, for it 
produces itself only in a certain number of cases, immunity 
may become acquired after a single subcutaneous injection 
more or less abundant of rabic virus. In this occurrence we 
enter in the order of phenomena on which the Pastorian 
method of preventive inoculation against rabies finds itself 
established. 

Though rabies is considered as a mortal malady, I do not 
think I am premature in sustaining that in a good number of 
cases after a bite, notwithstanding the introduction of the con- 
tagious elements in the organism and even in the nervous 
tissue, death is not produced, and the disease may pass unno- 
ticed. More than that, more or less serious symptoms of the 
invasion of the nervous system by the microbes of rabies may 



THE BACTERIOLOGICAL WORLD. 117 

manifest themselves without a necessarily fatal issue. Bour- 
rel ( * ) cites several observations of cure of rabies in ani- 
mals ; Leblanc, Jr., (f) has reported an observation of the 
same kind in the dog. A number of observations of rabies 
terminating in recovery have been also observed in man (I). 

It is not impossible then for the nervous system to free 
itself of the rabic parasites ; we understand easily, after this, 
the facility with which the cells of the cerebro-spinal system 
may resist the virus by means of methodical vaccination. I 
have demonstrated by my experiments on birds ( chicken, 
pigeon), that the microbe of rabies would not necessarily deter- 
mine incurable poisoning of the nervous cells. I have proven (§) 
that this class of animals contracted rabies rarely, but could 
transmit it in a mortal form to mammalians, proving that the 
former recovered spontaneously. In case when they appear on 
the point of succumbing when the quantity of virus injected 
has been too great, it is still possible to preserve them from 
death in sustaining their strength by artificial alimentation. 
Let me draw your attention, by the way, to the fact that birds 
may give a good experimental demonstration of acquired 
immunity, for as I have seen by experiments when they have 
once been attacked by and cured of rabies, it is impossible to 
make them contract this malady a second time. 

It was necessary, to produce the admission, and the 
understanding of the probability to render the organism 
refractory to rabic virus, to recall the facts that I have just 
explained. Let us now try and penetrate the principle of 
anti-rabic vaccination. Let us recall, in a few words, what this 
vaccination consists of: A man has been bitten by a rabid 
animal ; several weeks, several months and even several years 
may lapse before the accidents of rabies manifest themselves. 
Being given that we can accustom the animals to the attack of 
rabic poison, Pasteur's idea was to try and profit of the more or 

(*) Treatise on rabies ; Paris, 1874. 

( f ) Dictionary of medical sciences, article Rabies, page 128. 

(X) Lucas Benham — The Lancet ; March, 1890. In fifty observa- 
tions of human rabies terminated by cure, one-half, at least, appear to 
have been some very authentic cases of hydrophobia. 

(§) Experimental researches of rabies, Paris, 1884. 



118 THE BACTERIOLOGICAL WORLD. 

less prolonged delay, which separates the bite from the appari- 
tion of the accidents, to create immunity in accustoming the 
nervous system gradually to the strongest virus, before the 
morbid germ introduced by the bite of a dog has had time to 
invade it. I will not tarry to discuss the efficacy of the anti- 
rabic injection. I think that for all enlightened non-preju- 
diced minds the proof is made. Let us r pass, then, immediately 
to the examination of the conditions which render the cells 
capable of resisting the invading microbes in such a way as to 
repulse them victoriously. The remarks which follow may be 
applied to a great number of contagious diseases, against which 
we have been able, or may be "able in the future, to create 
artificial immunity. The first injections of the series of anti- 
rabic inoculations, according to Pasteur's method, are com- 
posed of liquids which are not virulent, that is to say, they do 
not contain any living microbes. This substance alone would 
probably be insufficient, at the dose ordinarily employed, to 
create immunity, and still we cannot say that it is absolutely 
necessary to obtain liquids containing the living microbe to 
create the refractory state for, as far back as 1886, Pasteur 
announced that he had been able to produce this refractory 
state in dogs, and guinea-pigs, to which he had inoculated 
nervous matter from rabid animals, and in which it had been 
impossible to reveal the presence of living germs by injection 
in the arachnoid of rabits. Does this imply that the refractory 
state may be produced, and is produced, chiefly by the soluble 
substances secreted by the microbe ? In the last years numerous 
examples have been furnished us of the refractory state by the 
aid of soluble substances secreted by germs ; I have just cited 
the experience of Pasteur on rabies ; I will recall, also, that 
Babes for the same disease ( * ) has obtained, in certain stages, 
the refractory state by the soluble substances alone. Immunity 
against bacteridian anthrax has been conferred to the suscepti- 
ble animals ( f ). ( J ) The soluble substances alone have given 

(*) Annals of the Pasteur Institute, July, 1889. 

( f ) Pasteur. Cornte Rendus of the Academy of Sciences, January 
30, 1888. 

( X ) Roux and Chamberland. Annals of the Pasteur Institute, 
August, 1888, 



THE BACTERIOLOGICAL WORLD. 119 

the same results in the hands of Mr. Salmon against hog chol- 
era ; we find the same against septicemia ( * ), against symto- 
matic anthrax ( f ) against typhoid fever ( J ). We may admit, 
therefore, that in anti-rabic vaccination the soluble substances 
play an important role. I would not, however, admit the pre- 
ponderance of their role. However, let no one misunderstand 
my thought, I will say even that without the presence of soluble 
substance, in the particular case that interests us, there would 
be no vaccination possible, but that this vaccination is secured 
by the presence of the living germs which secrete those soluble 
substances. I will explain myself. When an emulsion of 
rabic virus is injected in the subcutaneous cellular tissue, a 
certain quantity of liquid containing chemical substances 
secreted by the microbes, is immediately absorbed. In this way 
the whole organism, and the nervous cells in particular, are more 
or less affected by the contact of these chemical principles, I 
have noticed that the persons who submit to inoculation are 
much more tired during the first four days of treatment and 
during the following days, though the points of inoculation are 
much less sensitive and much less irritated than at the end (§). 
During these first inoculations it is only those substances which 
are injected. The nervous cell, therefore, accustoms itself from 
the beginning to the products secreted by the microbes. When 
this living microbe is in its turn inoculated it is at first put in 
contact with a cellular tissue irritated by the preceding inocu- 
lations ; this cellular tissue is accustomed to the secretions of 
the rabic germ, and it is probable that, at the point of inocu- 
lations, it suffers a shock which weakens it, and, when it is 
carried by one way or another to the central nervous cells, not 
only it finds them prepared for battle because they are already 
in a degree accustomed to poisons which the microbe secretes 
and the latter is weakened, but also very likely because a cer- 
tain proportion of the soluble substance elaborated by the 
microbe has remained in the organism and that the presence of 



( * ) Roux and Chamberland. Annals of the Pasteur Institute, 
December, 1887. 

( f ) E. Roux. Annals of the Pasteur Institute, February, 1888. 
( X ) Chantemesse and Widal. Annals of the Pasteur Institute. 
( § ) This is equally trua as I have observed in myself. 



120 THE BACTERIOLOGICAL WORLD. 

this substance is unfavorable to the development of the germ 
which has produced it. Let us add finally, that the cells of our 
organisms are in some sense microbes living collectively. 
Their secretions, like those of other microbes, are injurious to 
them when they are not eliminated ; but before they are elimin- 
ated they oppose, in a certain measure, invasion by external 
microbes. If it were not thus there would not be a single 
microbe in the world that would not be pathogenic. This may 
.account for a limited refractory state, but it does not explain 
to us how this immunity may last several years. We must con- 
fess that the theory of retention, that Prof. Chauveau has 
especially defended, is scarcely tenable with the exception of 
in the few days which follow inoculation, or the first attack of 
the malady. There is another theory, according to which, the 
microbe of a special affection could not develop itself again in 
an organism where it had previously produced a disease, and 
this because of the destruction, forever, of certain principles 
indispensable to its development. This theory of immunity 
does not appear to me more acceptable, for what would remain 
in the organism which would have been affected by a large 
number of contagious maladies if each of these maladies 
destroyed a " principle " of it? The experiments of Mr, Metsch-« 
nikoff show us well the manner in which certain cells of the 
organism battle against invading microbes, but I do not think 
that we must consider the white globules, phagocytes, the 
unique and indispensable agents of immunity. In the war of 
the cells they represent, so to speak, only the scouts ; the bulk 
of the army remains stationary, and guards its pontoons. I have 
already written it on several occasions : In the production of 
immunity several causes are instrumental ; among these causes 
some or others predominate according to the disease. In rabies 
immunity begins by the soluble substances which train the 
nervous cells to resist the toxic products, and then by the 
secreting germs of the toxic products which come, already 
attenuated, in contact with the cell ; but this does not explain 
how this immunity continues during a more or less extended 
period. To have a correct idea of the principle of immunity 
in general, we must leave the narrow limits of mechanism, 
and see in the nervous cells, as in all the other living cells, a 



THE BACTERIOLOGICAL WORLD. 121 

microcosm of man, as man is a microcosm, a reduction of the 
universe. The cell is constituted by three fundamental prin- 
ciples : it is formed of organic matter, and it has its soul, that 
is to say its portion of vital energy, and it is endowed with a 
proportional intelligence. Among the psychical qualities of a 
cell we find memory, an automatic memory I will concede, but 
memory, nevertheless ; and it is by the aid of this faculty that 
it remembers the processes of resistance that it has employed 
against this or that germ, the vital vibrations of which were 
inharmonious with its own, and it is without trouble that it 
returns at once to the same processes without giving to the 
parasites time to develop themselves at its expense, and to 
acquire strength for another attack in the future. This state 
of immunity may, however, be arrested for a time ; the cells 
may find themselves in a moment of vital depression, they 
may be surprised anew, or the period of time elapsed may be 
too great, and immunity is thus forgotten. 

In conclusion, I think that the mechanism of anti-rabic vac- 
cination does not differ perceptibly from that which produces 
the other immunity : the habit (accotitumance), or, if it is pre- 
ferred, the training of the cells is produced first by contact of 
the soluble substances secreted by the microbes before it is put 
in direct contact with these. In other cases of vaccination the 
soluble substance is put in contact with the cells when the 
microbe which secretes it is in their presence. As to the prin- 
ciple itself of immunity against rabies, it does not differ 
from that which directs its evolutions and assures its perennity* 
in other maladies ; in my opinion it is a phenomenon of cellular 
memory. 



122 THE BACTERIOLOGICAL WORLD. 

HYGIENIC. 



THE PRODUCTION OF IMMUNITY WITH THE CHEMICAL 
SUBSTANCES FORMED DURING THE GROWTH OF 
' THE BACILLUS OF HOG CHOLERA. 



BY E. A. V. SCHWEINITZ, PH. D. 

Chemical Laboratory, Bureau of Animal Industry, Department of 
Agriculture, Washington, D. C. 

As a continuation of the preliminary paper on the ptomaines 
from the hog-cholera germ, presented by us to the Chemical 
Section of the American Association for the Advancement of 
Science, in August, and published in The Medical. News, Sep- 
tember 6, 1890, we present now a somewhat detailed account 
of the successful experiments in the production of immunity 
in guinea-pigs which have been made up to date. The work 
from this standpoint again is of course a practicalcontinuation 
of the experiments of Drs. Salmon and Smith, made upon 
pigeons in 1887, in which sterilized culture-media were used 
for preventive inoculation. * * * * 

As to the name which should be given to the ptomaines 
and albumins from the hog-cholera culture-liquids, until 
their chemical constitution is more thoroughly studied, 
it would seem best, as there are several distinct swine 
diseases, to call the ptomaines from the hog-cholera germs, 
as a class, sucholotoxins, and the ptomaines which appears 
to be the principal factor sucholotoxin ( from the Greek 
2v$, a hog, XoXepa, cholera, from XoXrj, bile, and toxikon, 
poison). Sucholo-albumin would seem to be sufficiently 
distinctive for the albumin of these culture-liquids. As 
Hankin* shows, the name toxalbumin is hardly the correct one 
to apply to these substances. We shall, therefore, refer to 
the ptomaines and albumin by the names given above. 

The first of our experiments that we will record were made 
with sucholotoxin. 



^British Medical Journal, 1889, p. 810. 



THE BACTERIOLOGICAL WORLD. 123 

Experiments 1, — Two guinea-pigs, each weighing about 
three -fourths of a pound, were treated with a solution of about 
0.05 gram of sucholotoxin each. The solution was intro- 
duced under the skin of the inner side of the left thigh. 
Immediately after the operation the animal appeared uncom- 
fortable, but was not made ill. For a few days there was a 
rise in temperature and also a slight swelling at the point of 
inoculation, which, however, disappeared in about five days, 
and the animal was then well. 

Two more guinea-pigs were now selected as checks, 
approximately of the same size and weight as those which had 
been treated, and the four animals were then inoculated with 
0.1 cc. of hog-cholera virus each (0.1 cc. beef-infusion and 
peptone culture one day old, plus 0.2 of sterile, normal salt 
solution.) This is the dose which previous experiments made 
in the Bureau had shown to be the proper quantity to kill a 
guinea-pig in from eight to ten days. The inoculations with 
the virus were also made subcutaneously in the thigh. The 
checks died in eight and nine days. Post-mortem examination 
showed a large swelling at the point of inoculation, infiltration 
of a purulent, grayish substance into the connective tissue, and 
necrosis of the superficial layer of the muscles of the thigh. 
Enlargement and reddening of inguinal glands. Peyer's 
patches enlarged and pigmented ; liver pale and covered with 
a number of necrotic foci ; spleen very much enlarged, dark- 
colored and friable. Cover-glass preparations from the spleen 
and liver showed hog-cholera germs. This was the character- 
istic appearance of all the check guinea-pigs upon post-mortem 
examination, and it will not be necessary to repeat these 
details. 

Of the animals which had been first treated with the sub- 
stance mentioned, and afterward inoculated, one died two 
days after the last check. Autopsy revealed the following : 
At the point of inoculation in the left thigh the subcutaneous 
tissue was infiltrated with a grayish-white substance, and the 
superficial layer of muscles over the inner- side of thigh, and 4 
square centimetres of the abdominal wall were necrosed. Liver 
pale. Spleen much enlarged, dark colored and friable. 



124 THE BACTEKIOLOGICAL WORLD. 

Cover-glass preparations from the spleen showed a large num- 
ber of hog-cholera germs. Both ventricular walls of the heart 
were light grayish and very brittle ( necrosed ). The other 
guinea-pig of this set was quite ill for ten days, with a large 
swelling at the point of inoculation. This finally opened and 
healed, and the animal was quite well within three weeks after 
the inoculation, and has continued so to date — five months. 

Experiments 2. — The next series of experiments were 
made with sucholo-albumin from beef-infusion and peptone 
culture-media. 

Two guinea-pigs were again selected and treated with 
about 0.008 gram each of sucholo-albumin. There was a 
slight rise of temperature in the animals and the formation of 
a small, hard lump at the point of injection. This disappeared 
by the eighth day and the animals were quite well. Two more 
guinea-pigs were now taken as checks, and all four animals 
were inoculated with 0.10 cc. of hog-cholera culture. The 
checks died within seven days. The post-mortem appearances 
were practical^ the same as those noted in the first series. 
The two guinea-pigs which had been treated with the sucholo- 
albumin died ten days after the checks. This indicates consid- 
erable resistance to the disease. Autopsy showed, at the point 
of injection with the albumin, the subcutaneous tissue thick 
and reddened. The animals were considerably emaciated . At 
the point of inoculation a cyst the size of a walnut, and com- 
posed of a grayish, purulent substance, was also found. The 
muscular wall surrounding this was sprinkled with punctiform 
hemorrhages. Peyer's patches swollen and pigmented ; 
mucous membrane of small intestine covered with a dry, yel- 
lowish, firm layer of mucus ; stomach contained- a considerable 
quantity of liquid ; liver pale, and showed fatty degeneration ; 
spleen slightly enlarged and dark. Cover glass preparations 
showed no germs, but a culture made from the spleen showed 
hog-cholera germs. Beneath the peritoneum in the region of 
the spinal column, and in the mesentery was a considerable 
number of small grayish tubercles. Several other experiments 
were made by treating guinea-pigs with the albumin in varying 
quantities, all showing resistance, and subsequently immunity. 



THE BACTEKIOLOGICAL WORLD. 125 

Experiments 3. — Three guinea-pigs were treated with 
sucholo-albumm, 0.1 gram being given to each, subcutan- 
eously in the thigh. The albumin for two of the animals was 
derived from cultures containing blood-serum, the albumose 
given to the third was from ordinary beef-infusion peptone 
culture. Ugly ulcers formed at the point of inoculation, 
which healed, however, in from ten to fourteen days, and the 
animals with the exception of a slight rise of temperature were 
well. 

Two checks were again selected, and the five animals were 
inoculated with 0.1 -cc. hog-cholera virus. The checks died 
respectively in eight and ten days from hog cholera. The 
animals which had received the preventive treatment were 
slightly ill for a few days with swelling at the point of inocula- 
tion, which finally opened and then healed nicely, and within 
a week the guinea-pigs were well. 

Three weeks after the inoculation, one of these animals 
was chloroformed and examined post mortem. Not the slightest 
scar could be discovered; all the organs appeared perfectly 
normal, and no germs were found. 

Experiments 4. — Four guinea-pigs were treated, two 
with a mixture of sucholotoxins, two with sucholotoxin and 
albumin. The injections were made as before, subcutaneously 
in the thighs, and at intervals extending over a period of 
four weeks. The sore caused by each injection was allowed to 
heal before the next one was made. After the animals had 
recovered from the last treatment two checks were selected, 
and the six were each inoculated with 1-10 cc. hog-cholera 
virus. The checks died, one in eight and the other in ten 
days, the post-mortem examination showing characteristic hog 
cholera lesions. The animals having the preventive treatment 
were ill for about four days; those that received only the 
sucholotoxins being more dull than the others. There was 
also slight swelling at the point of inoculation with the germ, 
which subsided in ten days, after which the animals were per- 
fectly well, and have remained so — four months. 

( To he continued.) 



126 THE BACTERIOLOGICAL WORLD. 

SCIENTIFIC GLEANINGS. 



A NEW POISON IN CHEESE. 



BY VICTOR C. VAUGHN, M. D. 

During the past two or three years, we have received at 
the Hygienic Laboratory of Michigan University a number of 
samples of cheese, which, it was claimed, had caused nausea 
and vomiting in those eating of them, and in which we were 
unable to detect tyrotoxicon. Some of these samples produced 
vomiting and purging in cats and dogs to which the cheese was 
fed directly. Of course, the evidence that these samples had 
been the actual cause of the sickness among the people who 
had eaten of them was confirmed by the experiment upon the 
animals ; but, inasmuch as we were unable to detect the poison, 
we were compelled to report as follows : 

" The poisonous character of the cheese has been proven 
by experiments upon animals, but we have failed to demonstrate 
the nature of the poison. Tyrotoxicon could not be detected." 

One sample of this class was found by my assistant, Dr. 
Novy, to be very poisonous. Some of this cheese was covered 
with absolute alcohol, and, after standing in a dish for some 
weeks, the alcohol was allowed to evaporate, then 100 grams of 
the cheese were fed to a young dog, and caused its death within 
a few hours. Sterilized milk, to which, a small bit of the 
cheese was added, after standing in the incubator at 35° 0, for 
twenty-four hours, became so poisonous that 100 cc. introduced 
into the stomach of a full-grown cat caused death. Dr. Novy 
made plate cultures from the cheese and from the spleen and 
liver of the dead animals, and succeeded in identifying one germ 
as common to both. Sterilized milk inoculated with a pure 
culture of this germ, and kept in the incubator, proved fatal to 
cats. But with the advent of cold weather the germ lost its 
toxicogenic properties, which were not restored by subsequent 
cultivation in the incubator. 

In a second class of samples, the poisonous character of the 
cheese was not confirmed by direct feeding. Cats, rats and 
dogs were fed with the same quantities as above, without any 



THE BACTERIOLOGICAL WORLD. 127 

appreciable effect. The report made upon the samples was as 
follows : 

"Animals fed upon the cheese were not affected. Tyrotox- 
icon could not be found. The sickness of the people was 
probably due to some other cause." 

The last sentence of this report was probably wrong, as will 
be shown from the following experiment : Two kilograms of a 
cheese of this class were extracted repeatedly with absolute 
alcohol. The part insoluble in alcohol was then extracted with 
water. The aqueous extract, after filtration, was allowed to 
fall slowly into three times its volume of absolute alcohol. A 
voluminous, flocculent precipitate resulted. After twenty-four 
hours, the supernatant fluid was decanted, and the precipitate 
was dissolved in water and reprecipitated with absolute 
alcohol ; then it was collected and speedily dried on porous 
plates. A small bit of this precipitate was dissolved in water, 
and forty drops of this solution, injected under the skin on the 
back of cats, produced, invariably within one hour, vomiting 
and purging. After the partial collapse which followed the 
vomiting and purging, and which was evidenced by the animal 
sitting with its chin resting on the floor, recovery gradually 
followed. The same amount of the solution injected into the 
abdominal cavity of white rats rendered the animals, within ten 
or fifteen minutes, perfectly limp, and the only evidence of life 
observed was rapid respiratory movements. The rats lay upon 
their sides, and could be handled without manifesting any 
attempt at movement. In this condition, some died after three 
or four hours, while others, after lying in this position for from 
eighteen to twenty-four hours, gradually improved, and, after 
some days, seemed to be wholly recovered. 

This substance belongs to the so-called poisonous albumens. 
From its aqueous solutions it is not precipitated by heat or 
nitric acid, singly or combined. Its solutions respond to the 
Biuret test. It is not precipitated by saturation with sodium 
sulphate, nor by a current of carbonic acid gas ; therefore, it is 
not a globulin. It is precipitated by saturation with ammonium 
sulphate ; and this fact removes it from the peptones. 

That animals were not affected when fed with the whole 
cheese may be explained by the supposition that they did not in 



128 THE BACTERIOLOGICAL WORLD. 

this manner get enough of the poison to affect them. It cannot 
be said positively that the samples of cheese of the first class men- 
tioned above owe their poisonous properties to this substance. 
We have not had the opportunity of testing samples of this 
class since the recognition of the poisonous albumen in those of 
the second class. Four samples of the latter have been tested 
for the poisonous albumen, with positive results. 

It may be found that traces of this poison exist in all 
samples of green cheese. This point will be investigated. 

It is highly probable that the poisonous effects of some 
samples of sausage and meat are due to similar products of 
bacterial activity. — Medical and Surgical Reporter. 



Action" of living blood on bacteria. — Prof. Bonome 
has recorded the results of his researches on the following 
points : Whether physiological alterations in the blood play 
any part in modifying its destructive action on bacteria ; 
whether it is possible to produce the alterations in the compo- 
sition of the blood of such a nature that the normal, inimical 
action against bacteria may be altered ; and whether it is 
possible to derive any reliable data that will throw light on the 
subject of immunity. As a result of his experiments he comes 
to the conclusion that staphylococci introduced directly into 
the blood are destroyed in from ten to twenty-five minutes ; 
more rapidly in the blood of young rabbits than in older 
animals of the same species. He then, by injecting the poison 
obtained from the pus of an old empyema or a chronic abscess 
in small quantities into healthy rabbits, proved that the bacteria- 
destroying activity of the blood is increased, the organisms used 
being staphylococcus aureus, albus and citreus. He holds, 
however, that the introduction of such poison does not appear 
to exert any influence upon the similar activity of the fixed 
tissues. Poison from acute pus obtained in a similar manner 
appears to exert not the slightest^ influence on the destructive 
action of the blood ; while, owing to its effect upon the tissue 
elements, it diminishes their power of destroying such organisms 
as the staphylococci above mentioned. Similar poison from 



THE BACTERIOLOGICAL WORLD. 129 

pyogenic staphylococcus cultures does not increase this destruct- 
ive power of the blood against the above-mentioned organisms, 
and any immunity that is produced depends not on the rapidity 
and certainty with which the blood destroys the organisms 
introduced into its stream, but rather upon a greater resistance 
which the tissue elements exert against the bacteria poison, 
when they have become accustomed to the action of the poison 
by remaining in contact with the metabolic products of the 
same bacteria. He also gives experiments to show that water 
injected into the veins can diminish this destructive activity of 
the blood to a certain extent, but never completely ; for, 
although the animals so injected and control animals died 
about the same time, those in which water had been injected 
usually showed small purulent deposits in the kidneys and 
myocardium, and more or less fatty degeneration of the epithe- 
lium of the kidneys, so that he considers that, in addition to 
this slight diminution in the destructive activity of the blood, 
there is some alteration of the protoplasm of the cells, proba- 
bly due to the absence of salts and the cutting off of the full 
oxygen supply by the presence of water, by which their resist- 
ance is considerably diminished in certain areas, and owing to 
which they are more readily attacked by the injected staphy- 
lococci. — British Medical Journal — Journal of Medical Asso- 
ciation. 



130 THE BACTERIOLOGICAL WORLD. 

TECHNIQ UE. 



Staining tubercle bacilli. — Dr. M. Friedlander 
( Therap. Monat.) has devised a simple method of examining 
sputa for tubercle bacilli, by which two preparations can be 
completed in from five to ten minutes. The solutions necessary 
for this purpose, and which should be kept in readiness for use, 
are : First. Ziehl 's solution, viz., a 5-per-cent. watery solution 
of carbolic acid, to which an alcoholic solution of fuchsin has 
been added to the point of concentration. Second, A solution of 
5 grams of pure nitric acid in 100 grams of 80-per-cent. alco- 
hol. Third. A concentrated watery solution of methylene blue. 
The coloring process is as follows : The smallest possible 
quantity ( about the size of a pin's head ) of the specimen to be 
examined is taken up with a pincette or needle, and spread, by 
means of another clean needle, over the surface of a slide, in a 
thin layer about as large as a five-cent piece ; it is then dried in 
the air, during which time a second preparation is begun. The 
first one, when dry, is twice drawn slowly through the flame of 
a Bunsen burner or spirit lamp. It is next covered with 2 or 3 
drops of the fuchsin solution, and held over the flame, with the 
smeared side upward, until it begins to steam. It is then 
washed with water and covered with a few drops of the nitric- 
acid solution, which are allowed to act about half a minute, or 
until the color has quite disappeared ; then the acid solution is 
also washed off with water. A few drops of the methylene-blue 
solution are now added and allowed to act, without being 
heated, until the second preparation is equally advanced. The 
first preparation is again washed with water, then thoroughly 
dried with blotting paper and over the flame. It is now stained, 
and may be covered directly, without using a glass, with a 
drop of cedar oil, and microscopically examined. Two of these 
preparations will generally suffice for the investigation of a 
sputum. — Exchange. 

Staining bacillus tuberculosis in milk. — While milk 
is, doubtless, one of the most common sources of infection, in 
typhoid and in tuberculosis, from its very composition, unfor- 
tunately it is very difficult to demonstrate the presence of these 



THE BACTEKIOLOGICAL WORLD. 131 

micro-organisms in any given sample of the fluid. May's 
process of precipitation of the casein is very unsatisfactory, 
and in lieu thereof a writer in the Monitore dei Farmacisti 
suggests saponification of the fat globules by the following 
process : A drop of the milk is placed on a glass slip and two or 
three times its volume of a 1-per-cent. solution of sodium car- 
bonate is added, and the fluids mixed, with the aid of a plat- 
inum wire. The slip is then cautiously held over the flame of 
an alcohol lamp, or over the chimney of a kerosene lamp, and 
the liquid slowly evaporated to dryness. During the evapora- 
tion the butter particles are saponified, leaving a thin layer of 
desiccated soap on the slip. The subsequent treatment is 
identical with the older processes (staining with fuchsin, etc.). 
Rapid coloration with intense solutions of the reagents is 
preferable to the slower methods.* 

Kuhne's method of staining tubercle bacilli. — In a 
recent number of the Centralblatt fur Bakteriologie and Para- 
sitenhunde Kuhne gives a new method of staining tubercle 
bacilli devised by him to avoid the fallacies and difficulties 
which beset the examination of phthisical sputum. The main 
difficulty, according to him, is that of getting the sputum 
properly spread on the cover glass, and, to avoid this, he shakes 
up the material with an equal quantity of a concentrated solu- 
tion of sodium biborate. The effect of this is to render the 
material liquid and easily spread. The nummular matter from 
cavities he treats in alike manner, with a solution of ammonium 
carbonate, the latter being partially decomposed on heating the 
cover glass, a portion being volatilized. The subsequent steps 
are the same as in the old methods, heating of the cover glass 
over the lamp, staining with fuchsin, and bleaching with nitric 
acid and alcohol. The counter or ground stain suggested by 
Dr. Kuhne in place of methyl blue is a solution of picric acid 
in aniline oil. We have tried this method, and, while the results 
are not so good as those obtained by following Biedert's tech- 
nique, Kuhne's modification is so much simpler that it is to be 
preferred in all cases except those in which it is desired to 
make a ''show" or typical slide.* 



3 32 THE BACTERIOLOGICAL WORLD. 



/ 



Kultschitzky's eluid eor hardening and preserving 

MATERIAL EOR MICROSCOPICAL EXAMINATION. — The following 

is the method of Dr. N. Kultschitzky for the preservation of 
organic matter and its preparation for sectioning and staining : 

The fluid. — Supersaturate alcohol of 50° with a mixture of 
equal parts of pulverized potassium bichromate and cupric 
sulphate, and leave in absolute darkness for twenty-four hours. 
Decant, and keep in a dark place. Before using, add about 
1-4 of 1 per cent. (2.5 parts to 1,000) of glacial acetic acid. 

To use. — Put the object into this fluid and leave for from 
twelve to twenty-four hours, according to the size and density 
of the material. This must also be done in the dark. Transfer 
to alcohol of 95°, and in the course of twelve to twenty-four 
hours it will be ready for sectioning. 

In an article on the subject published in the Zeitsehrift 
fur Wissenschaftliche Mikroskopie some time ago, Kultschitzky 
gives the following general rules : For fixing the tissues it is 
important to use reagents that do not form tissue-like precipi- 
tates with protoplasm. This requirement is met by chromic 
salt, sulphate of copper, sublimate, and some other salts. 
Preparations in chromic salts, when transferred to alcohol, 
should be kept in absolute darkness ( H. Virchow), until the 
fixing reagent is removed so far as possible. 

All reagents which transform protoplasm into tissue-like 
forms, as, e. g., chromic acid, should be avoided, or their 
application must be controlled. 

Fixing fluids should contain an organic acid, e. g., acetic 
acid, which changes nuclein into an insoluble state. The acid 
must be used in a diluted form, as nuclein is dissolved in 
strong acid. The time of action must be short, as the long 
continued action of even a weak acid dissolves nuclein. 

It is desirable that the fixing fluid should contain alcohol 
in a small quantity, but strong alcohol dehydrates and induces 
changes in the protoplasm, and Kultschitzky thinks that, for 
conservation only, such fluids should be used as produce no 
further changes in protoplasm after it has once been fixed. As 
alcohol, Mueller's fluid and other fluids in. common use do work 
changes in the tissues. Kultschitzky recommends keeping pre- 
parations in either xylol, or toluol. — ^National Druggist. 



THE BACTERIOLOGICAL WORLD. 133 

QUERIES AND ANSWERS. 



Under this heading, we will answer, as briefly as possible, 
short, concise, rational questions of subscribers, concerning 
bacteriology only, as far as our knowledge may afford. 

Question. What is the best stain for bacillus of tubercu- 
losis in sputa, from a diagnostic standpoint ? 

E. M. D., Kansas City, Mo. 

Answer. The following is the method of Pittion and 
Eoux for staining bacillus tuberculosis in sputa, and is the 
method usually employed in our laboratory : 

Sol. "A." Ten parts of fuchsine dissolved in 100 parts 

of absolute alcohol. 

Sol. " B." Three parts liq. ammonia in 100 parts distilled 
water. 

Sol. " C." Alcohol, 50 ; water, 30 ; nitric acid, 20 ; aniline 
green to saturation. To prepare the last solution, dissolve the 
green in the alcohol, and then add the water, and, lastly, the 
acid. 

To use : Take of l ' A" one part and " B " ten parts. 
Heat until vapor begins to appear, then float the cover glass 
film down, which has been prepared in the ordinary way. 

One minute will stain the bacilli, but it is well to leave 
longer. Then rinse with distilled water and place it in solution 
" C " until the red color disappears, then wash thoroughly with 
distilled water and mount in Xylol balsam. 

It takes some practice to know just how much to discolor- 
ize. If the film of sputa is very thin, about 40 seconds in 
solution "C" will be sufficient. 

Zulil and Neelson's Method. — Float cover glass on carbolic 
fuchsine for 3-8 min. with heat. Decolorize in 25 per cent. 
HN0 3 , or H 2 S0 4 , then treat with 60 per cent, alcohol until 
only a rosy tint remains, then wash completely with dist. H 2 
and mount in Xylol balsam. 

These are the two methods that we have had the best 
success with, and, of the two, I prefer the former. 

There is a special, prepared stain, with directions on the 
bottle, known as " BurrilFs Stain. " This stain and method 
are excellent, but require a longer time. 



134 THE BACTERIOLOGICAL WORLD. 

Q. In your lesson in January number, you say that there 
is no spontaneous generation, but is there no such thing as 
sudden, new growth of living forms ? 

J. T. B., Little Eock, Ark. 

A. I mean that there never occurs spontaneous gener- 
ation, or spontaneous springing into existence, of anything 
endowed with life, except as generated by previously living 
beings. 

Q. I heard a discussion some time since, to the effect 
that Koch was not the discoverer of the contagiousness of 
tuberculosis. Is that a fact ? If it is a fact, who was the 
discoverer ? I wish you would kindly inform me. 

P. T., Hot Springs, Ark. 

A. From all I can gather, Villemin, of Paris, was the 
first man to demonstrate the contagiousness of tuberculosis. 
It was in 1865. But Laennec, in 1826, had noticed its 
inoculability by the accidental inoculation of the disease in 
his own hands in dissecting. Koch later, in 1882, discovered 
the germ to which contagion is due. 

Q. Seeing that you are practically interested in germs of 
disease, will you kindly tell me what system of filter is best to 
exclude them from drinking water ? 

A. So far as my experience goes, the system Pasteur- 
Ohamberland has all the qualities desirable. There may be 
others on the same principle equally good. The Pasteur- 
Ohamberland filter is on the market, and there is an agency in 
St. Louis. 

Q. Since you give lessons in bacteriology in the Bacterio- 
logical World, please tell me the best microscope to purchase 
for the study of bacteria. P. D., Kansas City, Mo. 

A. This is a delicate question, which we can answer only 
by past experience. In our laboratory, we use the foreign 
makes of Zeiss and Lietz, and the American instruments of 
Bausch and Lomb, and of Queen & Co. All these manufac- 
turers make splendid instruments, and we are much pleased 
with each of our instruments. There are, however, other 
excellent American or foreign microscopes in the country with 
claims to superiority, but, not having had experience with them, 
we cannot give an opinion. Write for catalogues to the 



THE BACTERIOLOGICAL WORLD. 135 

manufacturers or dealers, and get a description. As a rule, 
full explanations are given in them. See our advertisements. 

Immersion lens (oil in preference) are necessary. 

Q. I have read clippings and notices of Erlich's test for 
typhoid fever, but cannot find any full explanation of the 
subject. Will you kindly publish all information necessary for 
a practitioner ? 

A. We cannot answer this question as clearly and as fully 
in writing the reply ourselves as we can in publishing the 
excellent article of Dr. Simon, borrowed from the November 
number of the Johns Hopkins Hospital Bulletin, and which 
appears in March number. 

Q. Please advise me how to stain actinomyces. I have 
tried several ways without result. V. S., Indiana. 

A. In the first place, you will be disappointed if you 
expect to find the beautiful, regular, pear-shaped fungi joined 
in star fashion, which you may have noticed in certain books. 
The new methods of staining have revealed a more complete, 
and different, appearance. The actinomyces is a parasite 
formed by a club-shaped body terminating in a fine filament. 
It is somewhat like a whip with a short, stout handle and 
rounded, swelled butt. The butt end is usually encapsuled 
with a kind of cap, making its appearance still larger. In pus, 
the filaments and cap may be separated from one another, and 
may be stained separately. The finer, filamentous extremities 
stain blue or violet, and the encapsuled end stains red, as does, 
also, the cap itself, when separate. The fungi are usually 
arranged in rosette fashion, with the long extremity outward, 
and the filamentous extremities forming a dense, mycelial 
network centrally. But they may be free, or in groups of only 
two, four or five. It depends much on the source of the 
parasite. The filamentous extremity colors fairly by G-ram/s 
method, and the larger extremity by safiranine or eosine. 

Safiranine is prepared by Babes' method as follows : Make 
a saturated aqueous solution over heat, mix to 5 per cent, of 
aniline, and filter through a damp filter. 

Pus is spread on a cover glass in the usual way, dried, 
placed in this bath for several hours, and then treated with the 
iodized solution, clarified by aniline oil, and mounted in balsam. 



136 THE BACTERIOLOGICAL WOULD. 

This will color only the encapsuled head and the largest portion 
of the immediately-following filament. In coloring first the 
preparation with methyl violet and aniline blue by Gram's 
method, and then stain with eosine, both the finest central 
ramifications and the largest portions may be revealed, showing 
the connection between the two. 

The same methods are applicable to both pus and sections. 
A long practice is necessary to succeed fairly. 



FORMULAS. 



UNNA'S TREATMENT FOR RED NOSE. 

When red nose is due to acne rosacea Unna gives ichthyol 
internally in doses of 5 grains, and prescribes at the same time 
lotions of the same substance in aqueous solution. At night, 
the following paste is applied to the affected organ : 

3^ Sulphur 5- ss. 

Eice powder v 3 iv. 

Ointment of zinc oxide . . * \ . . . 5 v. 
Mix, and make a pomade. 

If the patient is scrofulous or debilitated, he also orders 
codliveroil, syrup of iodide of iron, etc. 

TREATMENT OF SYPHILITIC ULCERATIONS. 

Plumert gives the following applications for ulcers of 
syphilitic origin : 

3^ Mercury salicylate gr. xv. 

Potassium carbonate gr. xv. 

Distilled water , . . § vi. 

M. Dissolve. Sig. : Wet compresses with this solution 
and apply to the ulcerations. 

If an ointment is preferred, recurrence may be had to the 
following : 

1^ Mercury salicylate . . gr. xvj. 

Vaselin § j. 

Mix and make a pomade. 



THE BACTERIOLOGICAL WORLD. 137 

IODIDE OF POTASSIUM IN GONORRHCEAL RHEUMATISM. 

Kubinstein strongly recommends (in the Therapeutisclie 
Monatshefte ) the treatment of arthrites and inflammations of 
the mucous bursa? with potassium iodide after the following 
formula : 

1^ Potassium iodide 3 iv. 

Distilled water g v. 

Mix and dissolve. Sig. : Tabiespoonful every hour for 
two or three days. 

Amelioration usually begins after three or four doses. 
Kubinstein reports fifteen cases of acute and subacute gon- 
orrhceal rheumatism treated in this manner with the happiest 
results. — National Druggist. 

ULCERATIVE TONSILITIS. 

A favorite prescription of Dr. Wm. G. Moore, Prof, of 
the Principles and Practice of Medicine, Beaumont Hospital 
Medical College, St. Louis : 

1^ Katharmon § i. 

Sat. Sal. acid boracic 

Glycerine, aa • * . g i. 

M. Sig. : A teaspoonful in a wineglassful of hot water 
as a gargle in ulcerative tonsilitis and pharyngeal catarrh. 

Wm. G. Moore, M. D. 

SHANCROIDS AND ULCERATIONS. 

1^ Aristol 5 ]• 

Tarro-Petrolene ( Petr. Comp. No. 1 ) . g ij. 
Ft. ung. 
Sig. : Apply twice daily. 

Oz. Paqui^, M. D. 



138 THE BACTEKIOLOGICAL WORLD. 

LABORATORY REPORTS. 



PASTEUR INSTITUTE, PARIS. 
PERSONS TREATED WHO BECAME RABID DURING TREATMENT. 

Volvendey ( 12 years old ; of Beni-Saf, Algeria ). Was 
bitten by a dog, August 28, 1890, on the lower lip which 
carries two deep wounds, and on the left cheek, on which we 
count many superficial bites. None of these wounds have been 
cauterized. The animal was pronounced mad by Mr. Gorce, 
veterinarian. 

Volvendey was placed under treatment, September 7 ( nine 
days after he was bitten). The first signs of madness showed 
themselves September 24. He succumbed at the Necker Hos- 
pital September 28. 

Domenech, Jean ( 7 years old ; at Ille, Pyrenees- Orientales). 
Bitten by a dog September 6 ; first, on the right cheek, which 
made seven wounds, of which two were very deep ; second, on 
the upper lip ( a deep wound) ; third, on the left superciliary 
arch (a deep wound). The animal was pronounced mad by 
Mr. Volete, veterinarian of Perpignan. 

Treatment was begun September 10. Domenech took 
rabies September 28, he succumbed at the Children's Hospital 
October 2. , 

Vidalou, Sebastian ( 45 years old ; at Ille ). Bitten by the 
same dog as the preceding, September 6 : First, in the nose, 
which made a deep \found and many other superficial ones ; 
second, on the upper lip, a wound penetrating into the mucous 
membrane ; third, on the left arm, two deep wounds situated 
about the middle ; the clothes were torn. 

Treatment was begun September 10. Vidalou took rabies 
September 20, and succumbed at the Necker Hospital Octo- 
ber 3. 

The cauterization of the wounds was made with a red-hot 
iron 56 hours after the biting. 

Three other persons bitten by the same dog, very seriously 
on the face, were treated and are in good health. — Annates de 
VInstitut Pasteur. 



THE BACTERIOLOGICAL WORLD. 



139 



PASTEUR INSTITUTE. 



STATISTICS OF THE PREVENTIVE TREATMENT OP HYDROPHOBIA — SEP- 
TEMBER, 1890. 





J 


3 
1 

*4 

8 


B 

2 
6 

i 

24 
18 

*4 
16 
19 
30 
9 


6 
2 

23 
20 

i-9 

20 


C 




( s i mD ie 


*2 
4 

6 

'i 

8 

4 

11 

2 




Wounds of the head and face -J mu fJi D le 








1 

1 

2 








„, , „ , , , , ( simple 


5 


Wounds of the hands } mul P fci ple 




7 








Ineffective cauterization 


5 

7 


2 

4 








„_ , r ii i • i ij i( simple .' 


6 


Wounds of the limbs and trunk -j mu ]^i me 




7 


Effective cauterization > 






Ineffective cauterization 


4 

2 
6 


• • 








Effective cauterization 




2 


2 


Ineffective cauterization 










1 
1 
1 

2 




Not cauterized 


2 
1 

2 


24 




1 

78 
12 




Clothes torn 




Bites on naked skin 








T , , j French and Algeriens 




?.1 


lotai. . . -j strangers 




6 



Grand total, 141. 

Column A means the persons bitten by animals with hydrophobia 
and recognized experimentally ; column B, those bitten by mad ani- 
mals and recognized by the examination of a veterinarian ; column C, 
the persons bitten by animals suspected to be mad. 

The biting animals were : dogs, 123 times ; cats, 16 times ; donkeys, 
2 times. — Annates de Vlnstitut Pasteur. 



140 THE BACTERIOLOGICAL WORLD. 

REMEDIES AND CLINICAL NOTES. 

Beech wood creasote in tuberculosis. — This drug 
has recently taken the front rank as a remedy for tuberculosis. 
It certainly has achieved considerable reputation in that direc- 
tion, and numerous investigators are watching its administra- 
tion with great interest. A formula for its administration : 

I$* Creasote gr. xx. 

Alcohol 5 vi. 

Cinnamon Water § iii. 

Syrup 5 vi. 

M. Sig. : Teaspoonful three times a day. 

Diphtheria. — Dr. Slagle, m N. W. Lancet, says : My 
treatment for the past three years,, which has yielded such 
satisfactory results, has been as follows : 

3^ Calomel gr. xii. 

Soda Carbonate gr. xxxvi. 

M. ft. chart No. 12 ( or half this if young child ). 
Sig. : A powder every two hours. 

To be followed (when all used ) by 

1^ 01. Ricini 5 i. 

01. Terebinth 5 i. 

And every alternate hour, and until the case is fairly con- 
valescing, give a teaspoonful of a saturated solution of Merck's 
sulphite of sodium ; if the case is a bad one, direct a gargle or 
spray of the same to the throat every two hours. 

I do not claim that this is all the treatment that I now 
ever give for diphtheria, but I do say that this is the main and 
principal treatment, and upon which I confidently rely for spe- 
cific effects in removing the membraneous deposit from the 
fauces and preventing its reformation, often seeming to really 
abort the attack. My cases have all been fairly convalescing 
within three or four days on this treatment, with, of course, 
good support and that improved hygiene and regimen now well 
understood by every physician of ordinary ability. 

Aristol. — Aristol, a substitute for iodoform — have you 
used it ? It is a compound of iodine and thymol. I have 



THE BACTERIOLOGICAL WORLD. 141 

used it successfully in ulcerations, and have concluded that it 
is in many respects superior to iodoform, and when its action 
is fully understood will supplant iodoform in dermatological 
practice, and also in other conditions where indications call for 
the drug. 

Aristol is harmless ; no toxic effects are observed from its 
use. It is no doubt a powerful parasiticide, and in ulcerations 
of all kinds curative effects are more rapidly obtained than 
with iodoform or any other drug we have ever used. In 
diseases of the skin results from its use have been prompt and 
satisfactory in every respect. In eczema, herpes, psoriasis, 
varicose ulcers, lupus, dermatitis urticaria, inpetigo, contagiosa, 
etc., have been cured by its use. 

Aristol has the therapeutic properties of iodoform without 
its disagreeable odor and toxic effects. I use it as an ointment 
in vaseline — ten to twenty grains to vaseline, one ounce. It 
readily adheres to the skin. I frequently use it in the form of 
a powder, dusting it on ulcers, wounds, burns, etc. In ulcer- 
ation of the os, or abrasions of the vagina, it has often a good 
effect and is excellent. It can be used as an insufflator or in 
a powder-blower. It is soluble in fatty oils, but must be made 
in the cold, as heat causes its decomposition. It is insoluble in 
water and glycerine, sparingly in alcohol ; soluble in ether and 
chloroform. 

Aristol is a splendid remedy in the treatment of lupus and 
psoriasis. I know that in my hands it surpasses any other 
remedy in the cure of these two diseases. Several drugs have 
been recommended for lupus, etc. Ohrysarobin has been 
highly lauded for this purpose. Aristol is superior, and does 
not stain or discolor the skin, and cures more promptly. A 
good formula is this : Aristol, ten parts, olive or almond oil, 
sweet, twenty parts, lanolin one hundred parts ; dissolve the 
aristol in the oil and mix with the lanolin. .Bougies can be 
made with cocoa butter and two to ten grains of aristol. 
Pessaries can also be made in the same way for vagina or rectum 
to contain five to twenty grains of aristol. — Medical Summary. 

Iodo-carvacrol, a new antiseptic. — Oarvacrol is an 
isomer of thymol, and is formed from carvol which is found in 



142 THE BACTEKIOLOGICAL WORLD. 

oil of dill, oil of caraway and oil of mentha crispa. It will be 
remembered ( Circular for June, 1890, page 130 ) that a sub- 
stitution product is formed from thymol on treating it with 
iodine and potassium iodide in the presence of an alkali, to 
which the name aristol has been givec. On treating carvacrol 
in solution with iodine and potassium iodide and rendering 
alkaline, a yellow precipitate is formed which is analogous in 
composition to aristol save for the substitution of the carvacrol 
for the thymol. The new compound is described (Chem. Zeit.) 
as occurring in the form of a yellow-brown powder, which is 
insoluble in water, difficultly soluble in alcohol, and easily sol- 
uble in ether, chloroform and olive oil. It is unaltered by 
light, and when heated in a capillary tube softens at 50° C, 
and melts at about 90° C. to a brown liquid. From its compo- 
sition it would seem to be probably available for somewhat the 
same purposes as aristol. — Druggist Circular. 



ITEMS ON PUBLICATIONS AND OTHER 

MATTERS. 



THE STORY OF BACTERIA. 



BY T. MITCHELL PRUDDEN. 



This is an admirable, concise, compact exposition of the 
life, nature, properties and various special actions of bacteria. 
It is written clearly, plainly, and to the point ; every word having 
weight, and adding interest. It is just the work for all who 
are not versed in technicalities, and adds greatly to the 
library of medical men in general. General practitioners, 
particularly those remote from centers of medical education, 
could scarcely find any publication so well fitted to give in few 
chapters the most important part of the history of bacteria to 
the present date. 



DUST AND ITS DANGERS. 

Under this title, the same author has just issued, in the 
same plain, concise language, another interesting little volume, 
showing the dangers of germ-laden dust in and out doors. 



THE BACTERIOLOGICAL WORLD. 143 

The dissemination of bacteria in the air, their relative 
quantities, their action in the air passages, and, finally, the 
dangers of contracting consumption from the germs accidentally 
floating here and there, are graphically explained. 

It is a work that explains forcibly, in few words, and in 
language that will be plain to the unscientific, the hygienic 
principles that they may put in practice with much benefit. 
The physician, too, and even the scientific man will find much 
useful information. It is well illustrated. 

These two little books contain in a nutshell the most 
important part of the properties and history of bacteria. 



QUEEN'S OIL IMMERSION LENS IN PHOTO-MICROGRAPHY. 

We received some magnificent and clear photographs, made 
with one-fifteenth "Queen's oil immersion," by F. E. Ives and 
Dr. H. J. Detmers, of Ohio, whose reputation in this line of 
work is national. They represent the bacillus of anthrax at 
1,000; the bacillus of typhoid fever at 1,000; the bacillus 
of tuberculosis at 800, and amph. pellucida, lines 98,000 per 
inch, power 1,050 diameters. The latter was made by lamp- 
light. 

These photographs are among the finest it has ever 
been our fortune to see, and speak well for the lens and the 
operators. 



LA CLINIQUE. 



This medical periodical appears in a new and very neat 
dress in its first number for '91. It is quite an improvement 
on the old, both in style, color and fitness, and the magazine 
itself has made a step in advance. 

May the soul of this journal always prove as interesting to 
the mind as its ornament is pleasing to the eye. 



OLTJB RATES. 



\] 



m 



lllLi 




mn 



ti 



^ 



u± 



u 



LOGICAL WORLD. 



(ILLUSTRATED.) 
$3.00 PER YEAR IN ADVANCE. 



We offer it with the following publications at the rate indicated : 

The Journals American Medical Association 

( WEEKLY.) 
Price, $5.00. With Bacteriological World, $7.00. 



The Journal of Comparative Medicine and Veterinary Archives. 

EDITORS : 

W. A. Conklin, Ph. D., D. V. S., Director Zoological Gardens, N. Y. 

Rush S. Huidekoper, M. D., Veterinarian, Prof. American Veterinary 

College, New York. 
Price, S3. 00. With Bacteriological World, $5.00. 



THE AMEfjICAtiJETERipY REVIEW. 

Editor in Chief : Prof. A. Liautard, Principal, American Veterinary 

College, New York. 

Price, $3.00. With Bacteriological World, $5.00. 



THE CINCINNATI MEDICAL NEWS, 

Editor : J. A. Thacker, A. M., M. D., Cincinnati, Ohio. 
Price, $2.00. With Bacteriological World, $4.50. 



THE MEDICAL HERALD. 

Editor : Daniel Morton. M. D., St. Joseph, Mo. 

Price, $2.00. With Bacteriological World, $4.50. 



ST. LOUIS CL1NIQUE. 

Editors : Dr. William Porter and a Corps of Associates, 

St. Louis, Mo. 

Price, $1.00. With Bacteriological World, $3.75. 



Address THE BACTERIOLOGICAL WORLD, Columbia, Mo. 

Or for quicker attention: DR. T. J. TURNER, Mexico, Mo., 

Who has full authority to attend to all business matters for the 
Bacteriological World Company. 




DR. PAUL GIBIEF^. 






nn„ 




LUU 



u 




A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies. 

Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological Woeld, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to at. the 
same office. 

Authors of original articles published in this magazine will be entitled to twenty 
copies of the number containing it, provided, that they request the same in writing 
when sending the communication. Reprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. 

Discontinuation :— At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first of next January. 



COLLABORATORS 



Paul Gibier. M. D., New York. 
Prof. W. T. Belfield, RI. D., Chicago. 
Prof. A. W. McAlester, A. M., M. D., 

Missouri University Med. Dep't. 
Prof. Henry P. Loomis, M. D., 

New York . 



Prof. L. Bremer. M. D., St. Louis, Mo. 
J. W. Stickler, M. D., Orange, N. J. 
Prof. Paul Schweitzer, Ph. D., 

Missouri University. 
Prof. T. J. Burrill, Ph. D., 

Illinois University. 



B.T. 


Galloway, B. S., Sect 


ion of Vegetable Pathology, Washington, 


D. C. 


Vol. 1, 


COLUMBIA, 


10. 


Ui 61 Au 


MARCH, 1891, 


No, 3. 





EDITORIAL. 



STILL ANOTHER REMEDY FOR CONSUMPTION. 



Germany brings forth another discovery of an agent said 
to be remedial in tuberculosis. Prof. Liebreich is the dis- 
coverer. Drs. Fraenkel and Herman have experimented with 
it, and their results seem to indicate some therapeutical value. 

The remedy consists of e •' cantharidate of potash " ; it is 
administered by injection under the skin in very small doses, 
and its action seems to depend on an increase of transudation 
of serum (which we know is bactericide) and nourishing 
principles into the diseased tissues. Beginning with one-fiftieth 
of a milligram, the doses were gradually increased, during the 
experiments, to six milligrams ; the average quantity used, 
however, is two milligrams. 



146 THE BACTERIOLOGICAL WORLD. 

Dr. Heinemann gave the history of a number of cases of 
tuberculosis, some of them bad, which had been successfully 
treated. Hematuria appeared in certain cases after the injec- 
tion, but Liebreich did not consider this pathological phe- 
nomenon of much consequence, in view of the ultimate benefits 
to be derived in consumption. Trials are being made in cases 
of laryngeal tuberculosis and lupus of the head. 

Prof. Liebreich explains that his discovery was made public 
prematurely under pressure of Minister Yon G-ossler. This 
is very unfortunate ; the results of the premature announce- 
ment of Koch's remedy under similar pressure should, it seems 
to us, have taught more wisdom. Premature publicity of 
partly-made discoveries of such potentiality are injurious to the 
discoverer, the discovery itself, and only retards the benefits 
that may be in view, or even within reach. The people become 
more or less excited, are led by ignorance, confusion and intem- 
perate print to expect too much ; then, when accidents or 
failures occur, a depressing reaction follows, the discovery is at 
once classed as valueless, and the author suffers unjustly. 



NATURAL ANTISEPTICS OF THE BODY. 

The discovery of bactericide proteids of the animal 
body opens a new field of a very wide character, for the prac- 
tice of curative and preventive medicine in man and beast. 

It is well known that whilst certain animals are subject to 
a given disease others are not. For instance, cattle are liable 
to symptomatic anthrax (black-leg), but horses, man and 
certain other species are not ; horses and man are subject to 
glanders and cattle are not ; almost all animals are subject to 
charbon with the notable exception of rats. Prof. Hankin, being 
admitted into Koch's laboratory, was waiting in vain for an 
opportunity to learn the methods to prepare Koch's lymph. 
In the meantime he busied himself studying anthrax, a disease 
which had occupied his mind for some time. His first step 
was to find the reason for the natural immunity of rats against 
this disease. In this he was rewarded to the fullest extent. 



THE BACTERIOLOGICAL WORLD. 147 

He found that the body of rats contains a natural protective 
protein matter which kills the baccillus of anthrax, or at least 
prevents its growth. He succeeded in isolating this substance 
by making a glycerinated extract of spleen from rats, and pre- 
cipitating it afterwards with alcohol ; the protein is then 
soluble in water. The most minute doses, injected into mice 
suffering from anthrax, were sufficient to cause recovery from 
the most virulent form. 

In the calmest contemplation of this discovery, we must 
admit that it is extremely full of possibilities of the greatest 
magnitude. Almost every bacterial disease, if not all of them, 
find animals in which they have no effect, and "do not 
take." 

Mr. Hankin had some time ago announced the discovery of 
these natural protective substances ; now he has begun the 
practical application of them. It needs little thought, and no 
long discourse on anyone's part, to bring to one's mind the 
vastness of this achievement. 

Tuberculosis itself is a disease that some animals do not 
contract! So is Texas fever, in our country ; so is so-called. 
mad-itch in cattle ; swine plague, etc., etc. Besides that, it is 
a great step in advance in the settlement of the question of 
natural and acquired immunity. 



TREATMENT OF PULMONARY CONSUMPTION IN COLORADO. 

The Sanitarian, one of our most useful and worthy 
exchanges, publishes, in its February number, an exhaustive 
and very interesting article on the above subject, from the 
pen of S. Edwin Solly, M. D., M. K. 0. S. It is a presentation 
of the history, treatment and results obtained in 141 cases of 
pulmonary tuberculosis in the climate of Colorado (Colorado 
Springs). It shows that tuberculosis is curable, under proper 
climatic influences without any of the new discoveries ; unfor- 
tunately only a very small percentage of tuberculous people are 
able to visit this healthy climate. Physicians and the public 
would do well to read this excellent and instructive report. 
The results in those 141 cases are as follows : 



148 THE BACTERIOLOGICAL WORLD. 



First Stage. 

f Cured 36=58.04 per cent.^1 

I G. Improved*. 14=22.93 " >- Improved, 54=87.09 per cent. 

62 cases^ Improved 4=6.45 " j 

■ iSta 3 : v.':: : : : : til:* :: } Worse ' 8==12 - 90 per cent - 

Second and Third Stages. 

( Cured 11=13.92 per cent. ) 

G. Improved*. 14=17.72 " v Improved, 41=51.89 per cent. 

79 cases^ Improved 16=20.25 " ) 

iDeath. ".V WWZi^&M » } Worse - 38 = 48 ' 10 P er cent ■ 

Totals. 

( Cured 47=33.33 per cent . ) 

G. Improved*. 28=12.76 " \ Improved, 95=67.37 per cent. 

141 cases< Improved 20=14.11 ■• ) 

\^eSi:'':C:::. ; A^MM » [ Worse < 46=32.62per cent. 

*Greatly improved. 



GANGRENE OF THE LEG CONSECUTIVE TO ERYSIPELAS OF 

THE FACE. 

M. Reynier, before the Academy of Medicine, Paris, Jan- 
uary 21, reported an interesting case of gangrene following 
erysipelas, which suggests modification as to time of inter- 
ference in complications of this type. 

A young soldier, on the sixth of November last, presented 
" serious general symptoms, profuse sweating, diarrhoea, fever, 
followed after a few days by an adeno-phlegmon in the sub- 
maxillary region." During the month of December the left 
leg and foot were invaded by gangrene. In a few days the 
separation of the dead parts from the live tissues began, and 
Larry's amputation was practiced under the knee. 

The result seemed very good, but suddenly, a few days 
after the operation, the patient presented symptoms of a double 
infectious pneumonia, and nervous phenomena of the same 
nature. 

The moral of these rare complications seems to be earlier 
operation. 



THE BACTERIOLOGICAL WORLD. 149 

THE MICROBE OF EPIDEMIC CEREBROSPINAL 
MENINGITIS. 

Dr. Bonome investigated an outbreak of cerebro-spinal 
meningitis in Padua, and claims that he discovered a special 
micro-organism which is the specific cause of it. It is a strep- 
tococcus, differing morphologically from the various pathogenic 
cocci of this class, found by Nicolaier, G-uarnieri, Loemer and 
Weichselbaum ; it also differs in its preferences, in cultures and 
inoculation in small animals. 

Further investigations are necessary to verify these 
observations. 



INJECTION OF BLOOD SERUM OF THE DOG INTO THE 
TRACHEA AS A TREATMENT OF TUBERCULOSIS. 

Dr. Saint-Hilaire and Dr. Coupard, before the Society of 
Biology of Paris, January 31, gave the effect of intra-tracheal 
injections of blood serum as a treatment of tuberculosis. It 
had been discovered that blood serum raw was microbicide, 
and that its effect on the bacillus tuberculosis was in harmony 
with this discovery. 

Saint-Hilaire and his colleague began experiments with 
the view of finding the most inoffensive, less objectionable and 
yet the most absorbable channel for introduction and reception 
of the blood serum of the dog. In rabbits, dogs and man, in 
all of which intra-tracheal injections were made, the operation 
proved entirely inoffensive. Four cc. of serum were introduced 
at six different times in a man of twenty-two years of age, 
suffering from pulmonary and laryngeal tuberculosis, and no ill 
effects resulted ; not even a cough nor embarrassment in 
respiration. 

The results of these injections have been so far to improve, 
the general condition of the patients notably, and, even, the cure 
of a laryngeal case. It is to be hoped that this simple treatment 
may prove beneficial. 



150 THE BACTEBIOLOGICAL WORLD. 

ACTION OF BACTERIAL PRODUCTS ON INFLAMMATION 

In the course of 1890, Messrs. Charrin and G-amaleia sent 
a note to the Academy of Paris, calling attention to the prop- 
erty of certain microbe products in modifying inflammation. 
Certain experiments since bring the question back to us, and 
we, therefore, give the substance of this note. 

Buchner and Bouchard have established that, in the majority 
of cases, the intensity of the local lesion due to bacteria is pro- 
portional to the resistance of the inoculated animal. The rich- 
ness of this local lesion is furthermore usually due directly to this' 
animal resistance excepting in cases of absolute vaccination. 

With the view of explaining the presence or absence of the 
leucocytes, according to the state of immunity or receptivity, 
scientific men have thought that in the first case the white 
corpuscles were attracted by the bacterial secretions, or became 
accustomed to their action, — action which in the second case 
repulsed or paralyzed them. 

Mr. Bouchard had demonstrated that the injection of 
soluble products of some bacteria prevented diapedesis ; it 
restrained the migration of the leucocytes from the blood 
vessels. The following facts sustain this observation : 

If croton oil is rubbed on a rabbit's ear, redness and swelling 
occur within about four hours, and within eight hours considera- 
ble exudation and even blisters may be present. But if, after 
this oil friction, 4cc. to 8cc. of a sterilized liquid pyocyanic 
culture are injected every two hours until four or five or six 
injections are made, the symptoms above enumerated do not 
appear unless it be the next day in case the inoculation had to 
be discontinued because of their poisoning effects on the general 
system. 

We see that secretions of certain pathogenic agents may 
act on inflammation as a whole, prevent the congestion, the 
plasmatic transudations, and stop diapedesis. 

Investigations are now in progress with the object of decid- 
ing whether these agents act directly on the blood vessels, or 
through the intermediary of the nerves ; and also to find what 
other microbes, if any, secrete substances with such remarkable 
properties on inflammation. 



THE BACTERIOLOGICAL WORLD. 151 

PAUL OIBIER, 31. D. 

We have the pleasure to present with this issue an excel- 
lent likeness of one of our collaborators, Paul Gibier, M. D. 
(of the University of Paris), formerly interne of the hospitals of 
Paris ; assistant in pathology in the Paris Museum ; now 
director of the New York Pasteur Institute. 

This gentleman is one of the foremost bacteriologists of 
our time, and has achieved fame by knowledge and hard work. 
He has contributed a great deal to medical science since the 
year 1880. 

In 1884 he was intrusted by the French government with 
the study of cholera in France. In 1885, the same govern- 
ment sent him to Germany to study the nature of the bacterio- 
logical laboratories of the German Empire. In the same year 
he was sent to Spain to study cholera. In 1887 and 1888 he 
went in mission to Havana to study yellow fever. 

His numerous publications have appeared in French, 
Spanish and English. We refer our readers to notices and 
items in this magazine for the nature of the literature of which 
Dr. Gibier is author. 



THE BACTERIOLOGICAL WORLD. 

It is our aim to make this publication as useful, interesting" 
and as worthy in every respect as a magazine of its character 
can be. For the accomplishment of this purpose, we very 
earnestly request original contributions from those in authority 
to speak on the subject of bacteriology. 

The Bacteriological World circulates in every state, 
and in almost every territory in the union. It reaches, besides, 
Canada, and all European countries where any attention is paid 
to the question of which it treats. 

Writers, therefore, through this medium, may present 
their best thoughts, the results of their investigations, and their 
experience, to more widely scattered scientific and professional 
readers than through any medium of analogous order on the- 
continent. 



152 THE BACTERIOLOGICAL WORLD. 

LESSOJYS IN BACTERIOLOGY. 

LESSON THIRD.— PART I. 

phenomena Accompanying nutrition and vegetation of 

bacteria. 



We have seen that bacteria reproduce their like by spore for- 
mation and by fission. Let us now examine into their growth 
more closely. 

The spores, in relation to the adult bacteria producing them, 
are, as the grains of wheat in relation to the wheat plant itself ; 
as the apple seeds in relation to the apple tree ; the grape seeds 
in relation to the grape vine. In either case they represent the 
last resort of nature to reproduce the species. An apple tree 
may, however, be reproduced by grafting a branch on another 
tree ; a vine may be reproduced by planting a piece of stalk 
into the earth ; but these methods are not so natural as the 
forming and germination of seeds. 

In bacteria, however, the formation of spores and segmen- 
tation seem equally natural to the being in certain circumstances. 
Multiplication by fission occurs whilst there is nourishment, 
and the formation of spores, in the micro-organisms in which 
it has been observed, takes place chiefly when their nourish- 
ment is becoming exhausted ; then, bacteria form these seeds 
for the preservation of their kind, for the spores have more 
power of resistance than their producers. They are capable 
of germinating the moment they are generated, providing that 
they come in contact, at a proper temperature, with nutritive 
material that they can utilize and assimilate. But, when these 
conditions do not exist, they dry up, and may resist influences 
that would totally destroy their generators. For instance, if 
the spores of the bacillus of anthrax are kept in a dry atmos- 
phere, they will retain their germinating power for years, as 
some seeds of large plants in the same condition may. On the 
•contrary, if kept in moisture, warmth, and in contact with 
material from which they can extract nutrition, they will 
germinate, form new bacilli, which in their turn may throw off 
spores. 



THE BACTERIOLOGICAL WORLD, 153 

Therefore, schizomycetes may keep their species in the 
universe by forming spores which may resist for a long time 
the various influences tending to destroy. the species themselves. 
The surroundings and external influences to which the spores 
may resist vary a great deal. A spore may for a time, without 
growing, preserve its property of germination in the earth, 
water free from alimentary substance, and in various other 
things perhaps as well as in a dried condition. If any amount 
of nourishment exists, and the temperature is favorable, they 
vegetate, eat that up, and then the new individuals may form 
new spores. Certain disease germs are thus preserved and 
propagated. 

The spores resist a variable amount of heat, but all of 
them retain their vital energy in presence of much higher tem- 
perature than the bacteria. Thus the temperature of 212° F., 
boiling point, destroys in a few minutes the great mass, if not 
all kinds, of micro-organisms, but it takes more heat and a very 
much longer time at boiling temperature to kill their spores. 
Of course, the duration of the exposure to heat has a great deal 
to do with this destruction. 212° F. must eventually kill even 
the most resisting spores kept at this temperature a very long 
period, but I mean that whilst this degree of heat may kill a 
given germ in five minutes, say, it would likely require a half 
hour or more to kill its spores ; indeed, some resist 212° several 
hours. 

Experiments have demonstrated that in water and nutrient 
solutions, spores are killed in five to fifteen minutes at a 
temperature between 250° and 262° F. In a solution of gly- 
cerine, a temperature of forty or fifty degrees less, sometimes 
destroys their germinating power. 

On the other hand, freezing temperature is not sufficient to 
kill the germs ( or at least the great majority of them ), or their 
spores. Ice often contains living bacteria in abundance but 
usually they are in spore forms except in case of cocci. 

The most favorable temperature for the growth of most 
of bacteria seems to be between 86° and 104° F. At a higher 
or lower temperature, their vegetation and the formation of 
spores diminishes as the temperature increases or decreases, 
until all vegetation phenomena are arrested. 



154 THE BACTERIOLOGICAL WORLD. 

Microbes, notwithstanding their minuteness, have a very 
complicated substance. Their constituting material,- analyzed 
chemically, is found to contain carbon, oxygen, hydrogen, 
nitrogen and various mineral salts. Hence, they need nutrition 
that can supply these matters. 

The most suitable media of nutrition are vegetable and 
animal infusions, or broths. Thus it is apparent that albume- 
noid matters constitute proper food for the microbes. Never- 
theless, some germs at least, may live and vegetate at the 
expense of other matters, purely mineral salts of known com- 
position. Pasteur succeeded in developing the yeast plant 
(torula cerevisice) in a media free from any kind of albume- 
noids. It is thus that he defeated Liebig's theory of fermenta- 
tion. This great savant contended that fermentation depended 
on the destruction of the protoplasm of organized substances. 

Every process of vegetation has fixed limits of tempera- 
ture beyond and below which growth is arrested. This is just 
as true of bacteria as it is in the case of other organisms. 
Bacteria, in this respect, are as subject to fixed rules as are all 
other living things. Transgression of these limits may lead to 
destruction of the transgressors or temporary impairment of 
their vital energy. 

The bacteria being probably all plants, though there still 
exists some doubt about some of the microbes as I said in the 
first lesson, nourish themselves as vegetables. But being, as 
the fungi, deprived of chlorophyl, they cannot decompose 
carbon dioxide any more than the latter. The plants, with 
green substance, chlorophyl, operate thereby this kind of 
decomposition to nourish themselves, i. e., to extract and 
appropriate carbon. The fungi and bacteria must both obtain 
their carbon by utilizing organic carbon compounds already 
formed in other bodies. This is the case in all organisms 
deprived of chlorophyl. 

The thallophytes then, to which series of organisms the 
bacteria belong, can take up their nourishment only in a fluid 
or gaseous condition ; when they come in contact with nutri- 
tive substances they produce a transformation by means of 
diastases which brings about assimilable matters ; they find 
thus the means of nourishment. 



THE BACTERIOLOGICAL WORLD. 155 

A few bacteria contain a green substance which is thought 
by some authors to be chlorophyl. If this is the case, it is to 
be assumed that this particular kind is capable of decomposing 
carbon dioxide. It is needless to say that there is a change 
brought about in the material at the expense of which microbes 
exist ; all points to the universal physiological law of assimi- 
lation and nutrition ; viz. : First, the transformation of matter, 
non-assimilable as such, into assimilable substances ; second, a 
process of respiration of some £ind, either intramolecular, or 
by exhalation of more or less oxygen. 

All kinds of beings have their tastes and preferences for 
food. The fungi develop better in slightly acid substances ; the 
bacteria prefer slightly alkaline or neutral nourishment. 

According to their nutritive adaptation, bacteria are 
divided as follows : First, parasites, consisting of organisms 
feeding on living plants or animals ; second, saprophytes, 
microbes feeding on dead organic substances. But there are 
micro-organisms which go normally through their course of 
life as saprophytes, and have also the power of feeding partly or 
wholly as parasites ; these are termed facultative parasites. On 
the other hand, certain parasites, which normally exist as such, 
have also the property of living on dead organic matter ; these are 
termed facultative saprophytes. They are intermediate classes 
graded between the two extremes. 

PART II, 

DIASTASES. 

Part of what we have just seen proves that all the substances 
forming higher organizations enter into the constitution of 
bacteria, and our studies so far tend to show that nutrition 
depends, throughout the species of both (or all) kingdoms, on 
some process of digestion, respiration and assimilation governed 
by unalterable laws of nature. 

We have seen, too, in the beginning and in the course of our 
inquiries, that the first necessity is the digestion of food, within 
or without the organism, and that probably all beings may, for 
this purpose, secrete one or more substances capable of produc- 
ing the changes required to render the food assimilable. 



156 THE BACTERIOLOGICAL WORLD. 

We are given to understand that man, and I may say all 
complex animals, digest the solid and some of the fluid food 
swallowed before they can appropriate any of it to nourish their 
tissues; such is for instance the object of the diastase secreted 
in the stomach, pepsin. We have seen that complex plants 
may secrete diastases for xhe purpose of digesting, at certain 
periods, some inassimilable nourishment stored away in some 
parts of the vegetables for a special use, such as the sugar in the 
roots of the beet, which remains insoluble until the time of 
flowering, when a secretion occurs within the beet root which 
transforms this sugar into an assimilable kind, aud the plant 
absorbs and utilizes it. We have seen further that microbes 
have the same power. Let us now investigate the question of 
diastases a little more fully. 

Amylase is the name (in French) of a diastase which in the 
vegetable world digests starch, i. e., it has amylolytic proper- 
ties. This substance is secreted in a grain of corn (and some 
other grains) in process of swelling and germinating, under the 
influence of heat, moisture, etc., permitting the latent vital 
energies to develop. This diastase then gradually dissolves the 
starch at the expense of which the corn plant, at first embryonic, 
nourishes itself under proper conditions, until it has grown and 
acquired sufficient energy, i. e., physiological powers to begin 
the extraction of nourishment from its surroundings. 

Here is a clear process of digestion of starch by a plant before 
assimilation. Now, in animals what digests the starch of corn 
before its assimilation? It cannot be utilized as such. It is a 
diastase, that also has amylolytic properties, and which is 
secreted by the pancreas. 

If, now, microbes are fed starch, how can they assimilate 
any of it ? By a diastase, some sort of amylase secreted by them, 
and this is probably the secret of the action of the saliva on 
starch in the mouth. The saccharine substance formed there 
is probably due to various microbes of the buccal cavity, or con- 
tained in or mixed with the food masticated, Pasteur thinks 
microbes useful in digestion. Let us consider other kinds of 
diastases. 

Rennet ferment, trypsin, pepsin. — The first of these 
ferment principles exists in the stomach of the young calf in 



THE BACTERIOLOGICAL WORLD. 157 

lactation to the exclusion of other digestive agents. It is this 
agent that causes the coagulation of milk. Gradually, as the 
young begins to ingest other kinds of food, this principle 
disappears and is replaced by pepsin; finally the latter alone 
exists. In this state the ingested milk is also coagulated in the 
stomach; but it is by the action of the acid in the gastric juice 
and not by the rennet ferment. Thus, the milk ingested 
curdles in the stomach, both during the period of lactation, 
and afterwards in all stages of life, but by the action of 
different agencies. (The bovine species only furnishes a fit 
example of the digestion of milk in mammalians.) 

After this coagulation has taken place, however, neither of 
the diastases that produced it, in the young or adult, can act 
further upon the coagulum that they have formed respectively. 
This coagulum passes into the intestines; there it is dissolved 
and rendered assimilable by the action of trypsin. 

Pepsin, then, which in the presence of the acid in the gastric 
juice, acts as solvent of albuminous substance, is not capable of 
liquefying into an assimilable state the curd which, mixed with 
the gastric juice, it has brought about. Now among microbes, 
we find some that may nourish themselves at the expense of the 
caseine of milk just as in complex animals, and for that 
purpose they also secrete diastases that may be compared to the 
rennet ferment and the trypsin. 

If, for instance, sterilized milk is inoculated with the 
ferment microbe known as tyrothrix tenuis, there is first coagula- 
tion, then a dissolution of the curd without the advent of new 
germs. This, at first thought, seems to indicate an action 
contrary to that of the digestive ferments of the milk in animals, 
in which the stomach ferment principles cause a coagulum 
which they cannot dissolve, but which must be acted on for this 
secondary result by the trypsin of the pancreatic juice. It is 
not so however. It only indicates that the microbe, being a 
unicellular organism, has to do everything for itself, whilst the 
animal in the high type mentioned is formed of cells, among 
which labor has been divided and specialized as we have seen 
before. For the digestion of milk the cells of the stomach 
secrete certain substances; those of the pancreas, certain others; 
the individual microbic cell secretes them all. 



158 THE BACTERIOLOGICAL WORLD. 

If now we consider the mass of microbes as plants, we may 
think it difficult to reconcile these facts with what we know of 
the nutrition of vegetables of a complex character. The 
properties of these diastases would seem to indicate animal life 
instead of vegetable. This is only seemingly so. Digestive 
ferments of this order have been found even in complex plants, 
such as the carnivorous kind known as nepenthes, darling tonia, 
drosera, etc. These secrete substances capable of digesting 
albumenoids and even live animal beings. The plant known as 
carica papaya secretes (after Wurtz) a juice that is capable of 
digesting febrine in presence of an acid or in neutral media. 
Besides, if we recall to our minds the differences in the 
vegetation of large plants and thallophytes, owing to the presence 
of chlorophyl and specialization of labor in the cells of the 
former, and the absence of chlorophyl and the unicellular organ- 
ization of the latter, we shall see further explanations of the role 
of these diastases in bacteria considered as vegetables. Indeed, 
by a close study of the processes of nourishment of all kinds of 
living beings, no matter to what kingdom thev belong, we may 
harmonize the apparently inconsistent discoveries and facts con- 
cerning their respective properties. 

PART III. 

FERMENTATION. 

Mr. Duclaux admirably defines fermentations in the fol- 
lowing words : il Chemical transformations of substances dis- 
solved under the influences of organized beings, always deprived 
of chlorophyl, which develop and live in the interior of the 
liquid which ferments." This refers, seemingly, particularly 
to the well-known and common phenomena of fermentation in 
liquid substances ; however, such transformations take place 
in semi-fluid substances, and in more or less solid matters, but 
the changes in these were not supposed to be varieties of fer- 
mentation until the special properties of bacteria were clearly 
demonstrated. 

In the study of fermentation, Pasteur discovered that 
bacteria differ greatly in regard to their respective want of 
oxygen. Everything else being equal, he found at one end of 



THE BACTERIOLOGICAL WORLD. 159 

the scale, microbes that are favored in the highest degree 'by 
the greatest amount possible of atmospheric air containing 
free oxygen. Such is the case, for instance, with the bacillus 
subtilis, which grows so well in open air, in hay infusions, etc. ; 
at the other extremity, he found some favored by the exclusion 
of free oxygen ; such is the bacillus butyricus, which may 
develop in the very center of a mass of butter, and produce 
butyric acid, which, with other elements, contributes to pro- 
duce rancidity in butter. The first kind has been termed, 
therefore, aerobiotic ; the second, anaerobiotic. 

Between these two extremes, there are some requiring less 
pressure of oxygen than that afforded by atmospheric air. 
According to Nageli, the aerobiotic forms may also vegetate 
without free oxygen. Thus it would seem that this division of 
bacteria into aerobiotic and anaerobiotic is not absolute or 
defined by a clear line. 

.It is not our province to study all the phenomena of vari- 
ous fermentations here, but it is well to understand the most 
important ones. Therefore, we will touch on one or Wo 
more ; namely, the production of heat and certain results of 
fermentation. In our study of the nutrition of microbes, we 
have glanced at most of the other interesting ones. 

During the processes of fermentation, there is produced 
more or less heat. Everyone knows of the high temperature 
created in a manure heap, vegetables in a silo, or a stack of 
hay in fermentation ; it sometimes rises sufficiently high to 
consume to ashes, or even cause spontaneous combustion. 
" Fermentation," says Cornil and Babes, 'Ms a sort of intra- 
molecular respiration of the schizomycetes. The chemical 
reaction resulting then produces heat which is also necessary 
for the development and multiplication in the fermentescible 
matter." Robin has well explained that they are ''particular 
cases of nutrition, chemical nutritive acts, with production of 
heat as in most of such acts." 

Still, whether we consider fermentation from a botanical 
or chemical standpoint, everything is not absolutely clear, and 
it is possible that future investigations will modify the views 
now prevalent among naturalists, as to what individuals maybe 
properly termed ferment -producing agents. But we have a 



160 THE BACTERIOLOGICAL WORLD. 

mass of facts, proving at least the result of the existence of 
various microbes at the expense of fermentable substances, 
viz., transformation. One result of fermentation, or chemical 
transformation, brought about by the nutritive processes of 
micro-organisms, is important to know from a pathological 
standpoint above all others, that is, the possible death of the 
ferment agents themselves in their own products, poisoning in 
their own effete matter. Thus, in alcoholic fermentation, the 
schizomycetes causing it, yeast plant and others, are intercepted 
in their vegetation when the alcohol produced in the media in 
which they live has reached about 17 per cent.; the quantity of 
alcohol seems to constitute their poison. In the fermentation of 
wines in sealed vessels, fermentation stops thus by itself. If, on 
the contrary, this fermentation is allowed in the air, the surface 
will soon be covered by another organism which transforms the 
alcohol generated by the preceding germ into water and car- 
bonic acid ; this organism is the mycoderma vini, which burns 
the alcohol and transforms it into vinegar. A succession of 
microbic lives may thus exist and die in a given media, the 
first forming material fit, in a measure, for the nourishment of 
the second, the second for the third, etc. In these alcoholic 
and acetic fermentations, the first organisms acting were" 
anaerobiotic ; the alcohol resulted from yeast plants living at 
the bottom of the fermentescible matter, at a remote distance 
from the free oxygen of the atmospheric air ; the second or 
third forms that succeeded to the dead alcohol producers were 
aerobiotic, and lived on the surface, in contact with free oxygen. 

In pathology we find bacteria of the same aerobiotic and 
anaerobiotic classes. The bacillus of anthrax (charbon), for 
instance, belongs to the former kind, and that of symptomatic 
anthrax ( black-leg ) to the latter. In the cultivation of disease 
germs, we find, too, that they may be killed by poisons produced 
by themselves, and that different kinds of bacteria may succeed 
each other in the midst, and partly at the expense of the pro- 
ducts of preceding ones. 

Putrefaction. — The transformation of dead bodies of 
all kinds does not depend solely on one kind of germ but on 
several. The bacterium- termo, whilst it seemingly plays the 
most extensive and most important part in the decomposition 



THE BACTERIOLOGICAL WORLD. 161 

of animal bodies, has many associates in its life. Conse- 
quently, the phenomena known as putrefaction consist of the 
whole of the transformations ; putrefaction, in a word, is the 
sum total of the results of the nutrition of micro-organisms at 
the expense of dead vegetables and animals. In these phenom- 
ena " sugar is transformed into lactic acid, mannite, dextrine, 
glycerine, butyric acid, mucilage, etc. Alcohol is transformed 
into acetic acid, urea into carbonate of ammonia, albumen into 
peptone, or other similar substances. There also are generated, 
the putrid poisonous substance named sepsine ; the septic 
alcaloids of Zulzer and Sonnenschein ; the ptomaines found by 
Selmi, Gauthierand Breiger, etc.; certain narcotics ; leucine and 
tyrosine ; butyric, palmitic, margaric and fatty acids ; volatile 
products, iodol, phenol, scatol, sulphurated hydrogen, ammonia, 
carbonic acid, water," etc., etc. 

Therefore, the surface of the soil is the seat of very impor- 
tant changes ; many needed products originate there, without 
which, the plants of the earth must fail to find sufficient and 
proper food, Th<?re, ammonia is decomposed by microbes and 
nitric acid is liberated ; this acid in its turn may be decomposed 
into its formative elements or be returned to ammonia. These 
are the necessary • phenomena of the nitrification of soils. 
The action of bacteria in the superficial layers of the earth is 
said to be indispensable for the germination of seeds therein. 
Duclaux found that sterilized earth is improper for germi- 
nation. 

Saprogenous and chromogenous bacteria. — Among 
the putrefactive microbes, there are sorrie whose growth is asso- 
ciated with nauseous or aromatic agreeable odors. These 
bacteria are termed saprogenous. We all know that gangrene,, 
ozona, the secretions of the feet associated with the generation 
of certain organisms, present more or less disagreeable odors ; 
on the other hand, Babes and Galtier have cultivated micro- 
organisms which presented agreeable odors. 

The cliromogenous bacteria are those which produce certain 
coloring matters during their vegetation. The micrococcus 
prodigiosus, for example, produces a beautiful red ; the organ- 
ism of green pus produces a green color. In describing each 
bacteria, we will refer to them more specially. These 



162 THE BACTERIOLOGICAL WORLD. 

coloring matters may be separated by chemical processes, such 
as the action of chloroform, alcohol and ether. The action of 
heat, light, oxygen of the air, alters and fades these colors, and, 
under special conditions of nourishment, color-producing 
bacteria may be grown colorless. Such is the case if a little 
bichloride of mercury is added to the culture of the organism 
of blue pus. The growth of other chromogenous bacteria in 
similar media may also be without coloring substances, 

PART IV. 

PTOMAINES, LEUCOMAINES, TOXALBUMENS, BACTERIAL 

PROTEIDS. 

Under these names enter various products resulting from 
the growth of bacteria. 

Ptomaine is the name given to certain products of putre- 
faction ; chemically considered, they are found to consist of 
nitrogenous alkaloid bases of animal or vegetable extraction. 
They are probably produced both by independent bacteria, 
associations of bacteria, and, perhaps, some are chemical combi- 
nations of bacterial products of various kinds. Certain 
ptomaines are poisonous and others are not ; doubtless, products 
of the poisonous kind obtain greatly in various bacterial diseases 
more closely related to putrefaction processes, but they cer- 
tainly are not as universally dangerous factors in all bacterial 
maladies as was once thought ; for several of the pathogenic 
schizomycetes differ greatly in their transformation of tissues 
with the bacteria of putrefaction. There is a vast difference, 
for instance, between the results of the decaying of the lungs of 
a dead body and the results of the growth of the bacillus of 
tuberculosis therein during life, though both are due to bac- 
teria ; but there is not so much difference between the former 
phenomenon and infectious pneumonia ; and still less between 
the decomposition of a dead lung and a gangrenous living lung. 
In bacterial diseases, so closely allied to putrefactive processes, 
the ptomaines doubtless play an important role and may cause 
mortal poisoning. This is evidently the case in septicemia. 

Other substances, yet unclassified, have been found associ- 
ated with putrefactive processes, which resemble in their action 



THE BACTERIOLOGICAL WORLD. ] 63 

such poisons as coniine, strychnine, morphine, atropine, nico- 
tine, digitaline, veratrine, etc. ; some such matters were found 
in cadavers. These discoveries make very doubtful chemical 
analysis in certain suspected cases of poisoning. 

Ptomaines of a poisonous character have been found in 
canned meats, fish, oysters, in cheese, ice-cream, etc. 

Immunity against a disease is probably not produced by 
ptomaines, as was supposed. 

The following is a table of ptomaines, copied from 
Vaughan : 



164 



THE BACTERIOLOGICAL WORLD. 



TABLE OF PTOMAINES ( From Vaughan ). 



NAME 



Methylamine . . . . 
Dimethylamine . . . 
Trimethylamine . . 
Ethylamine .... 
Diethylamine .... 
Triethylamme . . . 
Propylamine .... 
Amylamine .... 
Hexylamine .... 
Tetanotoxine .... 
Collidine(?) . . . . 
Hydrocollidine (?) . 
Parvoline (?) ... 

Unnamed 

Ethylidenediamine (?) 
Trimethylenediamine 

Putrescine 

Cadaverine ..... 

Neuridme 

Saprine 

Unnamed ..... 
Methylguanidine . . 
Unnamed 



FORMULA 



Mydine . . . 
Neurine . . . 
Unnamed . . 
Choline . . . 
Betaine . . . 
Muscarine . . 
Mydatoxine . 
Mytilotoxine . 
Gadinine . . . 
Typhotoxine . 
Unnamed . . 






Tetanine . . 
Unnamed . . 
Tyrotoxicon . 
Mydaleine . 
Spasmotoxine 
Peptotoxine . 



OH 5 N . . 
C 2 H 7 N . . 
C 3 H 3 N . 
2 H 7 N . . 
C.H^N. . 
C 6 H 15 N. . 
C 3 H 9 N . . 
C 5 H 13 N.. . 
C 6 H 15 N. . 

C 8 H n N. . 

C 8 H 13 N . . 

C 9 H 13 N. . 

C 10 H 15 N . 

C 2 H 8 N 2 . . 

C 3 H 8 N 2 . . 

C 4 H 12 N 2 . 

O.H 14 N 8 . 

C 5 H 14 N 2 . 

C 5 H 6 N 2 , . 

C 7 H 10 N 2 . 

C 2 H 7 N 3 . . 

C 17 H 38 N 4 . 
C 8 H 1]L NO . 
C 5 H 13 NO . 
O.H^ffO, 

C 5 H 15 N0 2 

C 5 H 13 N0 3 

C 5 H 15 N0 3 

C 6 H 13 N0 2 

C 6 H 15 N0 2 

C 7 H 17 N0 3 

C 7 H 17 N0 2 

C 7 H 17 N0 2 

C 5 H 12 N 2 2 

C 14 H 20 N 2 O 4 

C 13H30 N 2°4 

C 7 H 18 N 2 6 



DISCOVERER 



PHYSIOLOGICAL 
ACTION" 



Brieger 

Nencki ....'. 
Gautier and Etard 

a a (( 

Guareschi and Mosso 
Brieger .... 



Non poisonous 



is 
a 
(( 



Poisonous 



a 
ti 

i i 



Morin 

Brieger .... 

Oser 

Oautier and Etard 
Brieger 



E. & H. Salkowski 
Brieger .... 






Poisonous 
Poisonous 






Non-poisonous (?) 

(?) 



Pouchet . 
Guareschi 
Brieger . 
Pouchet . 
Vaughan 
Brieger . 



Non-poisonous 

Poisonous 

Non-poisonous 

Poisonous 

Non-poisonous 

Poisonous 



Non-poisonous 
Poisonous 



Poisonous 



a 
a 

( ( 
a 



THE BACTERIOLOGICAL WORLD. 165 

Leucomaines should not be confounded with the ptomaines; 
they have some analogy, but are not identical. They have been 
defined as " basic substances found in the living tissues, either 
as the products of fermentative changes, or from retrograde 
metamorphosis." 

Some authors think that they are generated in the intesti- 
nal canal, and absorbed as assimilable matters. 

We gather, then, that they exist without disease, and that 
the most of them are non-poisonous. 

The leucomaines have analogy either with uric acid or 
creatinine; for this reason they are divided into two groups 
bearing these names respectively. The uric acid group is 
composed of adenine, hypoxanthine, guanine, xanthine, 
heteroxanthine, paraxanthine, carmine, pseudoxanthine and 
spermine. The creatinine group is composed of xanthocrea- 
tinine, amphicrcatinine, and three unnamed bases. 

There are undetermined poisonous leucomaines in expired 
air, vapors exhaled by dos^s, the saliva of man, the venom of 
poisonous snakes, urine, sewer air, etc. 

If the leucomaines are some of the products of fermentative 
changes, as is conceded, they may, of course, be produced by 
pathogenic microbes in the organism and thus both ptomaines 
and leucomaines may exist during disease. 

Bacigalupe, in making a study of immunity, came to the 
conclusion that it is produced by the means of leucomaines. 

Toxalbumose. — Eoux and Yersin have discovered, in pure 
cultures of the bacillus of diphtheria, a poisonous substance 
which is neither a ptomaine nor leucomaine, but apparently 
a kind of diastase, capable, by itself, of reproducing the general 
symptoms of the malady. Christmas discovered that the staph- 
ylococcus aureus ( a pus bacteria ) generates a sort of albumenous 
substance, which, when introduced into the anterior chamber 
of the eye of a rabbit, produces pus. Hankin, in Koch's labora- 
tory, isolated albumenous toxic substances from cultures of the 
bacillus of anthrax. 

These substances have not been identified with true 
albumen, but they have some analogy with it. Hence the 
name of toxalbumen, which has been given to them. Cornil 



166 THE BACTERIOLOGICAL WORLD. ' 

and Babes suggest the term of toxalbumose, which seems to me 
more proper. 

Brieger and Frankel have made researches concerning 
toxalbumose in cultures of various pathogenic bacteria, and 
found two classes, viz., one soluble in water, and the other 
insoluble in water. The toxalbumose of tetanus, that of anthrax 
( charbon ), and that of diphtheria are soluble in water ; the 
toxic albumens of typhoid fever, cholera and of staphylococcus 
aureus are insoluble in water. The discovery of toxalbumose is 
a most important one ; in connection with the knowledge of 
ptomaines and leucomaines, it advances medicine greatly 
towards the solution of preventive and curative treatments of 
many hitherto fatal, uncontrollable affections. It is prob- 
able that the so-called " Koch's lymph," for the cure of tuber- 
culosis, is of the nature of toxalbumose. 

Bacterial protein. — This is another active substance 
recently discovered in connection with bacteria. Buchner* 
Lauge, Roemer have demonstrated that there exists in the body 
the plasma of pathogenic bacteria, a chemical substance of 
protein nature, which has the property of producing disorders, 
and attracts the leucocytes in a marked degree. 

Injected under the skin it produces a " chemical " non- 
infectious inflammation accompanied with lymphangitis, which 
resembles the inflammation of erysipelas. This substance, 
being in the structure of the bacteria, is active only when set 
free by the natural involution of the organism or by their 
destruction by death. Though this discovery is recent, seven 
kinds of proteins have already been identified. The most 
virulent is that of typhoid fever. They may be obtained by 
treating pure cultures with weak solutions of potash (0.5 to 
1.5 per cent.), filtration, and then precipitations of the protein 
by acetic or nitric acid. 

*Centralblatt f. Chirurgie, 1890, No. 50. 

( Fourth lesson in April number.) 



THE BACTERIOLOGICAL WORLD. 167 

ORIGINAL ARTICLES. 



BACTERIA. 



BY ARTHUR G. HOPKINS. 



( Read before the Ontario Veterinary Medical Association, Ontario 
Veterinary College, January 27, 1891.) 

In bringing before your notice these organisms which 
have so revolutionized and stimulated theories regarding 
disease, its cause and cure, I do so, knowing that at present 
our knowledge regarding them is so incomplete as 'to cause a 
number of people to be skeptical as to their presence. Before 
going any further it is as well to know what are bacteria. 

Definition. — Bacteria are little masses of protoplasm, 
achlorophyllous, having a membrane composed of cellulose and 
mycoprotein, capable of multiplying by fission, hence their 
name schizomycetes, and require the microscope for their 
detection. 

History. — The first notice of them was over two hundred 
years ago, by Leuwenhceck, the father (?) of microscopy ; he 
found them in saliva, putrid water, fceces, etc. No particular 
notice seems to have been taken of them till Mueller, in 1773 
A. D., described them as animalcules (infusoria class), and, 
divided them into monas and vibrios. The next mention of 
them is by Ehrenberg, in 1838, followed closely by Schonlein, 
in 1839, who gave to the world "the achorion of Schonlein," 
the favus germ ; during this time Liebig, the great chemist, 
was fighting against the " germ theory " as opposed to the 
"chemical theory." In 1841, Duardin calls attention to them, 
and, in 1850, Davaine made the discovery, unknowingly, of the 
anthrax bacillus, mentioning at the time seeing threads in the 
blood of patients suffering from splenic fever ( pure anthrax ), 
although so near, yet he did not recognize this bacillus till 
thirteen years later. Up to 1853 they had been considered as 
animalcules, when the theory was advanced as to their being 
vegetables, this theory being converted into a fact by Davaine ; 
in 1859, he proved them to be vegetables. In 1862, Pasteur 



168 THE BACTEBIOLOGICAL WORLD. 

discovered the micrococcus ivrea ( the cause of ammoniacal 
fermentation in urine ), and in his experiments he found that 
boracic acid neutralized the effect of this germ. Dr. Guyot 
used this experiment, thus introducing the "era of antiseptic 
surgery." 1863 was memorable for Davaine's recognition of 
"the bacillus anthracis," and, in 1865, Lister, now Sir Joseph 
Lister, started in the work ( the perfection of antiseptic sur- 
gery ), which has made his name famous. Since then discov- 
eries have been made right on up to the present date, a cause 
being found for diseases previously mysterious. A few of the 
most important discoveries recently are : The B. mallei by 
Schutz and Loeffler in 1882 ; the actinomyces (ray fungus), 
B. tuberculosis by Koch. Investigations are being made as to 
the presence of germs and to determine their influence, whether 
pathogenic or not, in tetanus, influenza, cerebro-spinal 
meningitis, epizootic abortion, variola, contagious pleuro- 
pneumonia, hog cholera, canine distemper, malignant 
oedema, septicemia, texas fever maladie du-coit, and others too 
numerous to mention. The results at present are, in the 
majority, not entirely satisfactory as to immediately warrant 
the adoption of their causation by particular germs. 

By the earliest investigators, bacteria were considered as 
animalcules, — the reason for so doing was their property of 
motion • but further investigations proved their vegetable 
character, by optical tests with the microscope ; chemical 
tests : — with alkaline solutions, which would dissolve them if of 
animal constituents, and the use of stains ( methyl-violet, etc.), 
which immediately fixes well on most of the vegetables. They 
have no vital motion which includes the property of transla- 
tion, or movement on its own axis. Other methods of distin- 
guishing bacteria are by their spontaneity of motion, 
multiplication, more or less regularity of form, being unicell- 
ular, equality of dimensions in a given nutrition, and by 
various chemical tests. 

Classification. — The best and most generally accepted 
seems to be Oohn's classification ; he divides them according 
to morphology into four great classes. 

a. Sphero-bacteria ( micrococci ), occurring as round or 
oval forms. 



THE BACTERIOLOGICAL WORLD. 169 

b. Micro-bacteria ( bacterium ), slightly elongated and 
oval, with rounded ends. 

c. Desmo -bacteria ( bacilli ), cylindrical rods, rounded 
or square cut at their ends, 

d. Spiro -bacteria (spirilli), spiral shape and motile. 
They have also been classified according to spore formation 

into endosporous ( those developing withing the germ ), and 
arthrosporous ( those that reproduce externally ) ; but, as all 
bacteria do not reproduce by spore formation only, this classifi- 
cation is not generally used. Classification according to their 
oxygenation, negative or positive, and as to the effects produced 
by them, has also been tried. By this last they are divided 
into : 

a. Zymogenic, or ferment forming. 

b. Chromogenic, or color forming. 

c. Pathogenic, or disease producing. 

As we might expect, this last is the most important, and 
those in it occur as monads ( singly ), series, chains and zooglea 
(in which form a number of microbes are imbedded in a 
gelatinous or gluey mass). 

Reproduction and nutrition. — Bacteria reproduce 
themselves by fission or spore formation, the latter form being 
the most dangerous, as they resist stains and antiseptics better 
through their tough envelope. Growth in all forms is depen- 
dent on certain temperature ; many of them grow best at blood 
heat and will resist cold tolerably well ; electricity and sunlight 
affect many of them unfavorably. Moisture seems essential 
to all; other necessaries are nitrogen, oxygen and carbon, these 
all being taken in by endosmosis. They produce their 
effects by either : a. Using up the oxygen of the blood ; 

b, mechanically, i. e., owing to their presence and location ; 

c, by the formation of chemical poisons known as ptomaines, 
etc.; these are products peculiar to organisms, formed during 
putrefaction; some of them act as violent poisons. 

Modes of entrance into the system. — Bacteria may 
enter the body by the respiratory or alimentary mucous 
membranes, or by a wound in the skin ( normal epidermis is 
supposed to be impervious ) ; at present the theory held as to 
their introduction through the skin presupposes the existence 



170 THE BACTERIOLOGICAL WORLD. 

of some abrasion or injury. In the intestinal tract they may 
obtain entrance by the various channels of absorption. 
Bacteria have been found to be present in immense numbers in 
the alimentary tract from their presence in the voided excre- 
ment ; the various digestive juices would seem to have the 
power of neutralizing some of them to some extent. Experi- 
ments have been made (with, strictly speaking, negative results) 
to ascertain whether these organisms can obtain entrance 
through the skin, the B. mallei being made up into an oint- 
ment, and applied with friction. If organisms could readily 
enter a mucous membrane or the skin, the precautions of anti- 
septic surgery would be, generally speaking, useless. 

Spontaneity has been dealt with by Pasteur and Tyndall, 
as to render it unnecessary to say much about it. Like a good 
many new discoveries, there seems to be a danger to run " the 
theories," etc., to death ; to avoid this, Koch's postulates are 
to be adopted ; they are as follows : 

a. First find the micro-organism in blood, lymph or 
diseased tissue of the animal suffering or dead of the disease. 

. b. The micro-organisms, so obtained, must be isolated 
and cultivated outside the body and through successive gener- 
ations (in number, twenty or more), thus getting a pure 
culture. 

c. A pure culture thus obtained must, when introduced 
into a healthy body, produce the disease in question. 

d. In the inoculated animal the same organism must be 
found. 

These conditions fulfilled, we can say we have the history, 
etc., of any microbe. Often, however, we have to be content 
at present with the knowledge of their presence, without being 
able to fully determine their influence, whether pathogenic or 
not. 

The class, micrococci, does not include many pathogenic 
organisms compared with the bacilli. Among them are : 

a. Streptococcus erysipelatous, globular cells of small size,. 
and a tendency to grow in chains. 

b. Streptococcus pyogenes albus, said to be found in the 
abscess of strangles ; they resemble very much the strepto- 
coccus erysipelatous. 



THE BACTERIOLOGICAL WORLD. 171 

c. Staphylococcus pyogenes, etc., found in abscesses and 
suppurations. In their growth, they tend to form little grape- 
like masses. They are present in suppurative ostitis (vide 
Coats' Pathology). Micrococci are said to exist in diphtheria, 
but these do not have such a great interest to the veterinarian. 
They have also been found in the blood and lymph glands of 
patients suffering from rinderpest. 

Under the class bacterium, comes the " bacterium 
termo" the particular germ of putrefaction. It is short and 
appears as dumb-bells. The organism of fowl cholera is classed 
here by Klein, along with the bacilli by Coats. The B. 
diphther iticus, discovered by Loeffler, V. S., is supposed to 
be the cause of that dread disease, diphtheria. Doubts have 
arisen as to its action, it being found in some cases and not in 
others, and failing to produce the disease in question. 

The class, bacillus, is prolific in pathogenic organisms, 
and as such excites considerable interest. Bacillus of ulcera- 
tive stomatitis in calves shows the following symptoms 
(vide Klein, p. 126) : Ulcerations on the tongue and buccal 
mucous membrane ; the ulcer being a sore with overhanging 
edges ; wherever it touches it infects. The line of demarcation 
was found to be crowded with these bacilli. 

B . mallei are very minute, about the size of the B. tuber- 
culosis, shorter and thicker, are present in the lesions of 
glanders readily cultivated ( ? ), and easily communicated to 
susceptible animals. It is non-motile, and is communicable 
to man by accidental inoculation, causing the peculiar glan- 
derous symptoms, although these are sometimes hidden by 
some other affection (vide L. V. J,. Sept. 90). This bacillus 
has been cultivated through the various animals, such as 
rabbits ( ? ), mice ( ?), rats and guinea-pigs, the last giving the 
best results, and are available as a ready means of diagnosis, 
especially in chronic glanders ; the germ produces character- 
istic lesions in the ovary, vulva testes, etc., the male being the 
best for this purpose. The best and quickest method is the 
intra-peritoneal injection of the virus ; the lesions will be pro- 
duced in from two to four days. Coats says this bacillus loses 
virulence to a great extent by the fourth or fifth cultivation. 



172 THE BACTERIOLOGICAL WORLD. 

B. antliracis occurs as stiff, short and long rods ; according 
to Davaine, is slender and square cut at the ends ; it resembles 
the hay bacillus, and is said by Buchner to be capable of being 
changed into that .organism, or vice versa ; this theory is ably 
disproved by Klein and Koch. The bacillus (?) of symptomatic 
anthrax is shorter, broader and round at both ends. Klein men- 
tions that in true anthrax, the mother does not affect the foetus, 
but in symptomatic anthrax the foetus is affected. Pasteur has 
experimented with the bacillus anthracis, and has shown that a 
permanent vaccine is obtainable, ( Coats' Pathology.) 

B. tuberculosis is a thin, rod-shaped cell, shorter than the 
diameter of a red corpuscle ; it is often slightly curved or bent 
at an obtuse angle. The germ is the same in both man and 
animals, Klein to the contrary, however. This disease 
( tubercle ) infests some of the best bred herds in the world ; 
certain breeds being, seemingly, very susceptible to it. This 
bacillus is attracting a great deal of attention owing to Koch's 
discovery. It is said to be transmissible to man through the 
milk, and is thus a source of great danger. An experiment 
by M. Kichet in conjunction with M. Hericourt for the 
Biological Society, Paris, in regard to preventive inoculation 
for tuberculosis was successful (vide Lancet, Nov. 29, 90). 

B. of tetanus. —A germ said to be the cause of tetanus has 
been found in animals affected ; but at present facts are not 
forthcoming to warrant our stating conclusively that the germ 
found is the specific, or only, cause of the disease. 

Actinomyces (ray fungus). — This germ causes the 
disease in cattle known as actinomycosis ; the disease is 
transmissible to the equine and human species ; it is character- 
ized by induration of the tongue and other lesions connected 
with the jaw and face. In man there is a greater tendency to 
suppuration than in cattle. The disease was first recognized 
by Bollinger. The fungus consists of short threads arranged 
in a radiating fashion, many springing from one center ; they 
are united in the center by a matted mass of fibres, their 
arrangement thus accounting for the synonym, ray fungus. 
During our observations among the infectious diseases, we 
cannot help remarking that, no matter how seemingly virulent 
the disease is, some persons or animals are not attacked. This 



THE BACTERIOLOGICAL WORLD. 173 

immunity, for such it is, may be " natural/" e. g., anthrax 
attacks man, sheep and cattle, but does not attack dogs or cats. 
This immunity may be complete or incomplete, or it may be 
characteristic of a species, e.g., Algerian sheep are not prone to 
anthrax, and it has been said that strangles is unknown in the 
Arabian horse. When one per cent, are prone, and ninety-nine 
not prone, of a species to a disease, the condition is called an 
idiosyncrasy. 

Acquired immunity. — This immunity is conveyed to the 
individual in a variety of ways, i. e.: a. By having had 
the disease, e. g., strangles, b. By acclimation, e. g., malaria 
( this is not thorough immunity), c. By civilization (?). 
d. By age, e.g., typhoid fever rare in old people, e. By 
protective inoculation. 

Protective inoculation is of ancient origin, being practiced 
in India and the East long before known in England. Lady 
Mary Stuart Wortley Montague was the first to introduce it in 
Europe with the hope of mastering smallpox, which raged at 
the time. This noble project did not attain the object sought, 
as the virus being used to inoculate only spread the disease. 
Dr. Jenner took it in hand and perfected the scheme by his 
method of vaccination. He used a virus obtained from cattle, 
and the circumstances connected with his discovery are too 
well known to need recapitulation ; it is sufficient to say that 
he has immortalized himself. This discovery is not so much 
in itself to us as veterinarians, as is the principle emanating 
from it, showing, conclusively, if we cannot render a disease 
( or virus ) inert, we can at least prepare the animal organiza- 
tion to resist, with a varying degree of success, the inroads and 
attacks of these germs. Pasteur's work with the virus of rabies 
has achieved for him notoriety and a fair amount of success. 
Preventive inoculation for contagious pleuro-pneumonia is 
spoken favorably of ; the law of the United Kingdom prevent- 
ing any prophylactic treatment except slaughter has kept the 
practice of preventive inoculation back. In Australia and 
JSTew Zealand it has been adopted with success ; an outbreak in 
the last named place being stamped out by this means ( vide 
L. V. J., May, '90). 



174 THE BAOTEEIOLOGICAL WORLD. 

Koch's discovery of last year drew the attention of the 
enlightened Vorld, and, although it may fall short of the expec- 
tations, will no doubt encourage investigation in this direction. 
Pasteur located the virus of rabies in the spinal cord; by passing 
through monkeys, at intervals between the successive cultiva- 
tions, he obtained an attenuated virus. He experimented 
with the bacillus of fowl cholera and anthrax, and found that 
in eight days, at a temperature of 45°0, virulence was lost ; 
therefore, in less than eight days he had an attenuated virus. 
These experiments were subsequently verified by M. Ohauveau. 
Having thus given our attention to inoculation, and knowing, 
as we do, that these specific organisms must have a suitable 
nidus to produce their effect, the question arises, " What gives 
this immunity?" Immunity seems to be afforded, first, by 
the production of some chemical substance ( which may be 
found natural ) ; second, by the destruction of the food stuff on 
which these organisms exist ; third, is the result of a vital 
resistance.* 

Having mentioned the various prophylactic treatments 
internal to the body, we must now turn our attention to agents 
which will overcome these organisms when external to the 
body ; in a word, to antiseptics. No surgeon, or at the most, 
only a few surgeons in human or veterinary practice, think of 
going to work without these necessary adjuncts. Their inter- 
nal use has not been as uniformly successful as their external 
use, the reason being that in order to destroy the germ, the 
agent to be used would have an injurious effect on the body or 
organization to be protected. Experiments have been made 
successfully, however; it being found that minute doses of 
mercuric chloride fortified animals so as to resist the invasion of 
the anthrax bacillus. Sodium sulphite had the same beneficial 
action when administered to dogs injected with foetid pus. 
The majority of antipyretics are supposed to cause their bene- 
ficial effects by rendering inert the germ causing pyrexia. The 
chief antiseptics suitable for everyday use are : Carbolic acid, 

■ * Attention is directed to the article of Dr. Paul Gibier in January 
and February numbers, explaining immunity more fully ; also to the 
editorial in this number, concerning the discovery of protective 
proteids by Hankin. — P. P.] 



THE BACTEKIOLOGICAL WOULD. 175 

boracic acid and salicylic acid, iodoform, sulphurous acid 
solution, mercuric chloride, and creolin (Jeyes' fluid). This 
last-named drug is comparatively new on this side of the 
Atlantic, and is little known. It is derived from coal tar like 
carbolic acid, but it is far more potent as a germicide, antisep- 
tic, disinfectant and deodorant, while it possesses the great 
advantage of being free from all toxic properties; it is anodyne 
instead of irritant, and it is besides a haemostatic and styptic. 
It is alkaline in reaction^ and when treated with acids gives an 
oil, which does not give the carbolic-acid reaction with ferri 
perchlor. A two-per-cent. solution destroys the B. mallei, and 
has a restraining action on the bacilli of tubercle and anthrax. 
The influence and presence of bacteria is acknowledged 
every day by the use of antiseptics ; and if we examine the 
curriculum of any of the leading medical institutions of the 
day we find bacteria are there, either bodily or in the abstract. 
A knowledge of the actions and symptoms caused by these 
minute organisms being requisite for a correct diagnosis, which, 
when gained, gives us the key to treatment of a number of 
diseases, whether that treatment be curative or prophylactic. 



176 THE BACTERIOLOGICAL WORLD. 

HYGIENIC BACTERIOLOG Y. 



Report from the hygienic institute of Dr. R. Koch on the manner of 
producing a condition of Diphtheria immunity and Tetanus immunity 
in animals, by Dr. Behring, assistant at the hygienic institute at Berlin 
and Dr. Kitasato of Tokio, Japan.* 

[ TRANSLATED BY DR. A. C. BERNAYS & DR. W. A. BROOME, OF ST. LOUIS, MO.] 

During our long-continned studies on diphtheria (Behring) 
and tetanus (Kitasato) we have also approached the therapeutic 
and immunity aspects of the question. In both of these 
infectious diseases we have succeeded in curing infected 
animals, and also in so treating healthy animals that they cannot 
afterward become diseased with diphtheria and tetanus, 

The exact details of the methods of cure and immunization 
will not be given here; this subject will only be entered upon 
sufficiently to prove the following proposition: 

The immunity of rabbits and mice, ivhich have been immu- 
nized against tetanus, depends upon that property of cell-less 
blood-liquid {serum) to render harmless the toxic substances 
which are produced by tetanus bacilli. 

The explanation for the immunity, which has been 
expressed in the preceding sentence, has never been previously 
touched upon in any of the papers or works on the immunity 
question published up to the present time. 

Besides the doctrine of phagocytosis, which seeks to find an 
explanation of immunity in the cells, the bactericide or 
an ti- bacterial properties of the blood, and the power of the 
organism to become accustomed to poisons, were brought forward 
as explanatory elements. 

Whenever one of these three explanatory dogmas was proven 
insufficient, or was recognized as false by experimenters, the 
others were left to fall back upon per exclusionem. Bouchard, 
before the Tenth International Congress, in a most significant 
speech on the immunity question, defining the question, said : 
"Ne parlons done plus d* entrainement des leucocytes et 

* Detailed accounts will appear in the Zeitschrift fur Hygiene. 



THE BACTERIOLOGICAL WORLD. 17? 

d' accoutumance des cellules nerveuses aux poisons bacterieux; 
c'est pure rhetorique," and, " C'est en effet cet etafc bactericide 
qui constitue la vaccination on Y immunite acquise. " 

This positive statement amounts practically to the same as 
was said by Koger * in the following words : " La vaccination 
determine dans Torganisme des modifications chimiques qui 
rendent les humeurs et les tissues peu favorables a la vegetation 
ou microbe contre lequel on a premuni 1' animal. " 

Now comes one of us (Behring) and finds by his studies 
on diphtheria-proof rats and immunized guinea-pigs, that 
none of the above-mentioned theories can satisfactorily or in any 
way at all explain the immunity of these animals, and he was 
compelled to search for another element of explanation. After 
many vain efforts, the right direction was recognized when the 
poison-destroying effect of the blood taken from diphtheria- 
proof (dipththeria-immune) animals was found, and it was 
seen that the resistance and unreceptivity for diphtheria 
depended upon it. After the application of our experiences 
with diphtheria to tetanus, we arrived at results which,, so far 
as we are able to recognize, leave nothing to be wished for in 
the chain of evidence. 

The experiments prove the following propositions : 

First. The blood of rabbits that have been immunized 
against tetanus possesses properties which destroy the tetanus 
poison. 

Second. These properties have also been shown in extra- 
vascular blood, and in the serum of such blood. 

Third. These properties are of such enduring nature that 
they remain effective in the organism of other animals, so that 
we are able to achieve eminent therapeutic action by the 
transfusion of this blood or serum. 

Fourth. The tetanus-poison-destroying properties are 
absent in the blood of animals that are not immunized against 
tetanus, and, if the tetanus poison is injected into animals not 
immunized, the same may be shown to exist in the blood and 
other fluids after the death of the animal. 

We will now give the experiments which prove these 
propositions : 

* Contribution a L'etude de 1' immunite acquise, 1890. 



178 THE BACTERIOLOGICAL WOULD. 

A rabbit was immunized against tetanus after a method to 
be described hereafter by Kitasato in detail. After testing the 
degree of immunity in this animal the same was given lOccm. 
of a virulent pure culture of tetanus-bacilli. 0.5ccm. of this 
fluid sufficed to produce lockjaw in a normal rabbit and kill 
with absolute certainty. The first-mentioned rabbit remained 
healthy. 

The same had not only secured immunity against the infec- 
tion with living tetanus bacilli, but also against the tetanus 
poison; for it bore twenty times the amount of the poison without 
injury, which sufficed to kill normal rabbits in every instance. 

Some blood was taken out of the carotid of the immunized 
rabbit. Of this liquid blood before its coagulation 0.2ccm. 
were injected into the abdominal cavity of a mouse, and 
0.5ccm. were injected into another mouse. 

Both mice, together with two others, were inoculated with 
virulent tetanus bacilli after twenty-fours ; the inoculation was 
made so strong that the two control-mice were taken with 
tetanus after twenty hours, and died after thirty- six. 

Both mice which had been injected with the blood of the 
immunized mouse remained permanently well. 

The greater amount of the blood was left standing until 
the serum had separated from the clot. Of this serum, six 
mice each received 0. 2ccm. into the abdominal cavity; twenty- 
four hours afterward they were infected, but all six mice 
remained permanently well, while six control-mice died of 
tetanus after less than forty-eight hours. With this serum 
therapeutic successes have been achieved by first infecting 
normal animals and afterward injecting the serum into the 
abdominal cavity. 

Furthermore, we have made experiments with the serum 
which established the enormous poison-destroying effect of the 
same. 

A tetanus culture ten days old was prepared, which was 
freed of germs by filtering. Of this, O.OOOOoccm. sufficed 
to kill a mouse in from four to six days, and 0.0001 ccm. 
sufficed to kill a mouse in less than two days with absolute 
certainty. Now we mixed 5ccm. serum of the tetanus proof- 
rabbit with lccm. of the culture and allowed the serum to act 



THE BACTERIOLOGICAL WORLD. 179 

upon the tetanus poison contained in the culture for twenty- 
four hours. Of this mixture, four mice received 0.2ccm. 
(equal to 0. 033ccm. culture) an amount equal to more than 
three hundred times the quantity ordinarily a fatal dose for 
mice; the four mice remained permanently well. The control 
mice died thirty-six hours after a dose of O.OOOlccm. of the 
culture. The mice of all the experiments above-mentioned — 
those which received the serum in the abdominal cavity as well 
as those which received the mixture — have become permanently 
immunized ; they have withstood repeated inoculations with 
virulent tetanus bacilli without showing the slightest trace of 
disease. This fact is of particular importance, because in 
innumerable exeriments no mouse, no rabbit, in fact no animal 
ever tested, was found naturally immunized to tetanus, and also 
because no other method attempting to immunize any species 
of animals against tetanus has ever succeeded. 

We may, therefore, draw the conclusion that the conception 
expressed in the beginning of this article about the manner in 
which the immunity is brought about answers a far-reaching 
demand for a causal explanation. 



THE PRODUCTION OF IMMUNITY WITH THE CHEMICAL 

SUBSTANCES FORMED DURING THE GROWTH OF 

THE BACILLUS OF HOG CHOLERA. 



BY E. A. V. SCHWEINITZ, PH. D. 

Chemical Laboratory, Bureau of Animal Industry, Department of 
Agriculture, Washington, D. C. 

( Continued from February Number.) 

Experiments 5. — Six guinea-pigs were inoculated for this 
experiment, two with solution of thesucholotoxin and four with a 
solution of the mixed sucholotoxins. The sucholotoxin solution 
produced only slight local lesions, while the mixed toxins caused 
ulceration at the point of injection which did not heal for two 
weeks. The treatment in this case again extended over a period 
of from three to four weeks. The animals having by this time 
recovered, the test experiment with hog-cholera virus was tried 



180 THE BACTERIOLOGICAL WORLD. 

Four of the animals mentioned above were taken — two from 
each set — and also two checks, and the six were inoculated. 
The checks died in eight and nine days, the autopsies showing 
the characteristic conditions of death from hog cholera. Those 
that had the preventive treatment were ill and dull for from 
four to six days after the inoculation. At the point of inocula- 
tion there was also some swelling and infiltration, very slight, 
however, compared with the similar swelling on the checks. 
In the treated animals the swelling sloughed and healed, and 
within ten days after the inoculation they were perfectly well. 
To test the resistance of the animals, that had been treated by 
this method, to ordinary exposure the following experiments 
were conducted. 

Experiments 6. — Two guinea-pigs that had received the 
preventive treatment, two blanks, i. e., animals that had 
received no treatment, and two check animals that were 
inoculated with hog-cholera virus were placed in one large 
cage. The checks became ill and died in eight or nine days 
from hog cholera. During this time the cage was cleaned only 
three times, so as to give full and free opportunity for conta- 
gion. One week after the checks had died one of the blanks 
became ill, and died within ten days. The autopsy showed 
hog cholera lesions. The second blank became ill a few days 
after the first blank succumbed, and died within thirty days. The 
animals which had the preventive treatment are now and have 
been quite well, though continually exposed for five weeks to 
every opportunity for contagion. 

Experiments 7. — These experiments are a step in advance 
of those already recorded, and, although not quite so conclusive, 
indicate that the proper methods have been adopted. 

A pure chemical compound, prepared synthetically in the 
laboratory, was used for treating the guinea-pigs. Three 
animals were taken, and this compound was administered to 
them by the method already used. There was a slight rise in 
temperature of the animals and swelling and soreness at the 
point of injection. After this had healed these animals and 
two checks were inoculated with O.lcc. of hog-cholera culture. 
The checks died in eight and nine days. The animals which 



THE BACTERIOLOGICAL WORLD. 181 

had been previously treated became ill, two dying five and six 
days after the checks. The third entirely recovered. 

Post-mortem examination of the two that died showed the 
following : At the point of inoculation the skin had sloughed 
away over an area of 1 sq. cm. The superficial muscular layer 
was necrosed over an area of about 3 sq. cm. and to a depth of 
1 mm., lymphatics in the fold of the knee much enlarged ; 
Peyer's patches enlarged and pigmented ; spleen very slightly 
enlarged and not discolored ; kidneys reddened ; lungs normal. 
Cover-glass preparation from the spleen showed a few hog- 
cholera germs. On both sides of the spinal column were sev- 
eral grayish tubercles, from 1-4 to 2 mm. in diameter, lying 
just beneath the peritoneum. This material is being more 
fully tested, and experiments which promise to be successful 
are also being made upon hogs. Autopsies made from the 
animals of Experiments 7, three or four weeks after their 
recovery, showed that the parts were perfectly normal, not 
•even a scar being left upon the skin, and the immunity produced 
was therefore perfect. 

It is important to add that in all the experiments great 
care was taken that the solutions used were free from germs, 
cultures always being made. In cases in which the albumen is 
used this is particularly important. A single precipitation with 
absolute alcohol does not suffice to destroy the germs, and it is 
necessary to free the solution from germs by means of a Pasteur 
filter, or in some other suitable way. Therefore, experiments 
made with material which has not been tested for germs are 
practically of no value. As to the poisonous character of the 
ptomaines, a single large dose is sufficient to kill a guinea-pig 
in from one hour to two days. The autopsy of a case of this 
kind is as follows : Jjiver, pale and fatty ; subcutaneous tissue 
over abdomen, necrosed and infiltrated ; muscle, soft and 
friable ; other organs, apparently normal. 

The experiments here recorded show : 

First. That in guinea-pigs complete immunity from hog- 
cholera can be produced by chemical inoculation. 

Second. The sucholotoxins and sucholo-albumen are 
■equally effective in this respect, and a mixture of these two 
products gives greater immunity than either used by itself. 



182 THE BACTERIOLOGICAL WORLD. 

The effect of the albumen in producing immunity from anthrax 
has already been pointed out by Hankin, his experiments being 
very successful. 

Third. The sucholotoxins given in large doses produce 
death. To produce immunity they should be administered in 
small quantities at a time and at frequent intervals, the system 
being in this way accustomed to the poison and enabled to 
resist it. 

Further study in this interesting line of work is in pro- 
gress. 

The tabulated results of the foregoing experiments are 
appended : 



THE BACTERIOLOGICAL WORLD. 



183 



P 

i— i 
P 

o 



« 

- w 

p 

o 

B 
O 

o 

ffl 

c 



o 

P 
Q 
O 
« 



H 
P 

a 
3 
p.. 

H 
o 

CO 

H 

P 
co 
P 
03 

Q 
P 
E-i 
<! 
P 
P 
pa 
< 



















73 

in 






















CD 










>>J3 






CD 
PI 

ce 

CO 

'co 

© 

4-3 










S£h 


a 

'S 


>d 

CD 
P-i 
CD 

> 
O 

© 

CD 


>» >» 




e3 2 p*, 

^ 2 * . 
• m ^ 


73 
© 


t* 


S3 
© 


+3 
'p 


-*3 




+3 73 


1^§ 


4-3 

o3 

© 
f-i 

+3 


... 


fee 




a 




©eg 

© 03 g 
*3«, 2 


co 
03 


co 

>3 


i 
I 


2 
2 




2p 


.2 


-« 


03 

73 




. 71 




■"73 


o.2^ 


-u 






73 


73 




'C CL 


a © 








CD 


© 




CD — 


73 ^ 


3 


a. 


1—1 




s-i 




h'C 


co % © 


CO 




0. 


z 


© 




© y 


S"= > 


© 


73 




> 






° P 
° ec 


^73 O 


w 


CD 

,73 


73 
© 


c 
© 
© 


O 
© 
© 




■° ^ © 

*° ° CD 

0££ 




T—i 


s 


B 


K 




Kp: 


EH 


co cc 
















day 
ocu- 
viru 

; Of 
















«-l C r-i ^ CO 






c 




O 






0-SS--SJM 


OS 










Ol 


C3 OS 


mber 
tween 
ion wi 
nd dei 
chec 


73 




t3 




73 


'O 


73 73 


PI 




p 







p; 


a pi 


03 




- 




a 


c3 


03 03 


GO 


fc- 


oc 




30 


ao 


GO 00 


£ ©4-3 03 
















£-°J5 
















=4-1 

O 












P 
ec 


co 

© 


CD-* 












5 




CM! 


OJ 


(N 




0? 


,3 © 


c o 
















* 
















=4-1 
















o ■ 
















CO 
















mber 
nimal 
used. 
















O* 


CNJ 


CC 


C? O? IN O? OJ 


oo 
















P o3 
















fc 
















e3 
















i~"S - 
















hole 
i use 
eacl 
mal. 


6 

CD 


o 


c 


6 


6 c 


6 c 


6 


U (/J , .rH 


o 


-o 


73 


73 


^73 73 7; 


73 
















a^ 


O 














• 










a 








£3 


















0) 










2 








2 










p 








4-3 










£ 








e3 

CD 










13 








-t-J 




.2 


p 




73 


.2 P 
o c 








*4 
O 
et-c 

CD 
co 


o 


S 

13 


£ 

c 


1 
>< 

i 5 


C 


y 

P 


© 
> 2 


3 


4-> 






* 4-3 


+3 4- 




© 


o 


6 


: 


o o c 


OC 


> rfl 
















5 ° 


.5 


o 


o 


1 


) o 


c c 


o c 


"C 


r=J 


,0 


,c 


,0,0 ,£ 


,flr£ 


! © 


CD 


o 


© 




> © 


© e. 


© c. 


' 53 


4^> 










J C3 


- 


pi p 


5 P 


a 


02 


CC 


a 


1 COCO 


QQa 


2 P-i 


£ 








- 


<sV 


4 C* 




^•r 1 
















umbe 
expei 
ment. 


i— I 


hH 




i > 


> 


H 


H K 








H 1— 1 




t 5 


' t> 


K% 



















© 



73 

© 



p 



184 THE BACTERIOLOGICAL WORLD. 

SURGICAL BACTERIOLOGY. 



ANTISEPTIC SPONGES. 



prof. J. perrins (in Gaz. de Gynec). 
Take a string of fine, new sponges, neither too large nor 
too small. Commence by beating them with a small hammer, 
or a piece of wood, in order to knock out the dust and any 
mineral particles that they may contain. Examine particularly 
the point where the sponge was attached to the rock ; of ten there 
are minute grains of sand firmly adherent at this point, and 
which should be absolutely removed to avoid irritating an 
already painful wound. It is better to cut this point off with 
the scissors. Having done this, wash the sponges freely in 
water, squeeze, and place them in fountain basins, preferably 
enameled ones, containing the following solutions : 

Hydrochloric acid 10 grains. 

Water 1 litre. 

Allow them to remain six hours in this mixture, then wash 
freely again in water, and steep them for six hours in the fol- 
lowing : 

Water 1 litre. 

Permanganate of potash 1 gram. 

By treatment of a chameleon solution of a beautiful, 
intense violet hue, they will become more or less rapidly discol- 
ored, and, after washing freely in water, will retain a brownish 
shade. 

Immerse the sponges next in the following «liquid for two 

hours : 

Liquid bisulphite of soda 10 grams. 

Water 1 litre. 

Hydrochloric acid 1 gram. 

A slight odor of sulphurous acid gas will be perceived. 
After a little while the sponges lose their brown color, and 
assume a beautiful, whitish yellow shade. 

Wash again freely in water and introduce them into a pre- 
serving fluid made as follows, where they should be kept until 
needed : 



THE BACTERIOLOGICAL WORLD. 185 

Water 1 litre. 

Carbolic acid 1 gram. 

Alcohol 5 grams. 

Dissolve the carbolic acid in the alcohol and mix together 
in the aqueous solution. 

In order to remove the odor of the carbolic acid, which the 
laity always associates with hospitals, we may use in the place 
■of the above solution the following preserving fluid : 

Thymol . 1 gram. 

Alchohol 4 grams. 

Water 1 litre. 

— Reg. Pharmacist. 



SELECTIONS. 



THE TREATMENT OF TUBERCULAR JOINT-DISEASES BY 
MEANS OF IODOFORM INJECTIONS, 

By Prof. Tedor Krause, Halle. 
[From the Archiv. fur Klinische Chirurgie, Vol. 41,] 

[TRANSLATED FROM THE GERMAN BY H.W. LOEB, A. M..M. D., ST. LOUIS, MO.] 

In my communication, in the Berliner Klinische Wochen- 
schrift, 1889, No. 49, upon the above subject, I mentioned that, 
in my researches toward the cure of tuberculosis of joints by 
intra-articular injections of iodoform, I had chosen at first only 
the severe cases for the most part with abscess formation, those 
which our experience had shown could not be cured without seri- 
ous operative interference. When the result of the severe forms 
of the disease became encouraging, I further experimented 
with the milder cases and in the very beginning of the disease. 
In these we can with fair reason hope for a cure before the pus 
forms in the joint. I have treated the following joints, 
during two years, with intra-articular injections: 

Kneejoint 36 times 

Hipjoint 13 times 

Tarsaljoint 6 times 

Wrist joint 5 times 

Elbowjoint. ... 1 time 



186 THE BACTERIOLOGICAL WORLD. 

The treatment is not yet completed in a portion of these 
cases, or has not been carried on sufficiently long to offer a 
definite conclusion. As far as I could determine in regard to 
the cases treated before, a cure resulted in the 

Kneejoint 15 times 

Hip joint 4 times 

Tarsaljoint 1 time 

Wristjoint 3 times 

An improvement occurred in the great majority of those 
treated. These are the successes ; now I shall not pass by the 
failures. In a case of kneejoint disease, arthrectomy was 
necessitated on account of advancing tuberculosis — and in a case 
of tuberculosis of the -tarsaljoint, in a woman, aged . 45, an 
amputation of the lower part of the thigh was later performed. 
A man, aged 19, with tuberculosis of the posterior portion of the 
anklejoint, was taken sick with meningitis four weeks after the 
washing out of the abscess and the injection of the iodoform 
mixture, and died ten days later from tubercular meningitis and 
pulmonary tuberculosis. Whether or not the surgical 
interference was the immediate occasion for the outbreak of the 
tubercular meningitis, I do not venture to decide. The period 
of four weeks between the injection and the inception of his 
fatal sickness seems somewhat long. It is well known, 
especially since Kcenig has made observations upon this subject, 
that as an immediate result of operative interference, especially 
in the train of smaller operations, as scraping and the like, such 
as was done in the tubercular joints, acute miliary tuberculosis 
occasionally takes place. According to our experience, the first 
symptoms of the general injection occurs within ten or fourteen 
days, and not as in the above-mentioned cases after four weeks. 

A further case is that of a patient, aged 37 years, with 
severe kneejoint and pulmonary tuberculosis, who while the 
kneejoint disease was perceptibly benefited by. four injections, 
succumbed to advancing phthisis pulmonalis. 

I obtained the best results in knee and wristjoints, 
Especially were the results thorough in the severe cases of 
tuberculous suppuration of the wristjoint occurring in older 
people, which otherwise would have necessitated amputation of 
the forearm, or at least resection of the wristjoint, and they 



THE BACTERIOLOGICAL WORLD. 187 

were sometimes surprising, even when the patient had phthisis 
pulmonalis. So, also were two cases of the severest form 
of tuberculosis of the hand accompanied by suppuration, one of 
which was complicated by fistula; both who had been sent to 
the clinic for amputation were perfectly cured, and in both 
patients the movement of the finger is painless, although, of 
course, active motion of the wristjoint is impossible. Just so 
severe is a third case, a woman, who is now after three 
injections, on a road to certain recovery. 

Among the kneejoint patients, in a single very severe 
case a cure with free motion resulted. I may here exhibit 
some of these patients, so treated. This boy, Eeinhold R., 
had formerly, upon the head of the right tibia, a caseous 
sequestrum with the formation of a tubercular abscess. The 
sequestrum had been removed. At that time there was no 
disease of the kneejoint apparent. Some, months later the 
patient returned for treatment, with severe tuberculosis of the 
right kneejoint. There were great swelling of the capsule and 
a large purulent effusion with stretching of the ligaments so 
that side motion was warely possible. Five injections of ten to 
twenty grams (5ij ss to 3 V ) were given within a period of four 
months. Since then a year has intervened, and, you see, the 
healing of the condition has continued. The patient can bend 
his knee to the right, and is not hindered in the use of the leg. 

This boy, Walter F., three years of age, had no effusion 
but presented severe swelling of the capsule, and a slight 
subluxation backward, so that he had to submit to a triple 
extension. As the internal condyle of the tibia was greatly 
swollen and pressure was very painful, a deposit here could well 
be conjectured. A cure followed four injections, and it has 
remained well for a year. The boy can bend his knee to the 
right, but cannot fully extend it; but in this the use of the leg 
is not prevented. 

Finally, I present to you an eight-year-old boy, Max V., who 
suffered from a tuberculosis, of moderate severity, in the 
left kneejoint. A gelatinous, purulent discharge with cheesy 
shreds of tissue was constantly flowing from the opening. By 
three injections a cure with complete normal mobility of the 
joint resulted which still persists (a year later). 



188 THE BACTERIOLOGICAL WORLD. 

But not simply in childhood should the results be 
thoroughly exhibited, but also in the adult. 

So I present a woman, aged 40, who suffered from a left-sided 
gonitis (inflammation of the knee) with trifling purulent 
discharge and severe swelling of the capsule. By passive 
motion which gave the patient intense«pain, crepitation could 
be felt. In four weeks after the first injection the patient was 
entirely free from pain, and on standing could somewhat bend the 
knee. Three other injections caused a cure so that the patient 
can now bend her knee to the right, and for five months can go 
around without any sensation of pain. Slight crepitation can 
always still be obtained, though it is entirely painless. 

This man, aged 20 years, furnishes the best example of the 
value of the method. This was one of those cases of consid- 
erable and very soft fluctuating swelling of the capsule, which 
offer such a bad prognosis as to a cure. The young man had 
previously been treated with plaster-of-Paris dressing and with 
irrigation of a sublimate solution, but the swelling of the 
capsule increased, and the pain became intolerable. For a long 
time he was unable to stand or to walk. Each movement of the 
leg being^extremely painful; yes, the slightest shaking of the bed 
occasioned simply by approaching it caused the patient exquisite 
pain. Without morphine, he could not exist. Regarding this 
case I consulted fully with Von Volkmann who gave not much 
hope for a lasting cure even by an arthrectomy. He advised me 
in any case to prepare myself to make an amputation of the 
thigh which later would be necessary. He was treated with 
iodoform injections as a last resort. Its action in this case was 
astounding — I cannot call it anything else. The pains were 
really slight, only fourteen days after the first injection ; 
fourteen days later they had entirely disappeared ; after the 
second injection, and four weeks of treatment, the patient 
could go around without supporting apparatus. In all four 
injections of 20 to 40 grams (5v-x.) were necessary within 
the four months. The cure was followed by kneejoint anchy- 
losis, and now a year after remains the same. 

In all the cases of hipjoint disease, in which on account of 
its duration a final sentence was passed, I, nevertheless, obtained 
anchylosis. This concerns the cases with abscess formation, to 



THE BACTERIOLOGICAL WOELD. 189 

» 

which I applied the injection. Still, here I may show two, who 
recovered from coxitis of the severest form, with numerous 
abscesses, loss of cartilage and even having had the luxation- 
posture. We obtained a recovery with more or less well- 
maintained motion, where we applied the iodoform injections 
in the beginning of the malady. This point experience must 
decide. The cases which I have heretofore treated have not 
been observed sufficiently long to warrant a definite conclusion, 
especially two cases of coxitis with abscesses, which at first 
showed free movement but later resulted in complete anchylosis. 
Although anchylosis intervenes in the treatment of the hip- 
joint there should, however, be no ground to cast aside the 
method for this joint. 

It was the opinion of Volkmann ( and I properly verified 
this opinion at the previous congress ) that after a partial 
resection of the hipjoint a firm, bony anchylosis between the 
acetabulum and the resected femur with moderate abduction of 
the leg was the best thing for the poorer classes of society, who 
cannot bestow so much attention to the after treatment. Such 
an anchylosis alone will guard against frequent severe returns 
of adduction and flexion. The same opinion naturally holds for 
the cure of coxitis without resection. 

The treatment with iodoform injections is, therefore, easily 
carried' out while the patient, who is, of course, watched with 
the ordinary orthopedic measures, needs remain under observa- 
tion only a few days after the slight attack. 

We have frequently observed, where patients or their 
relatives had been properly instructed, a subsequent injection 
taken ambulant. Now, in whatever the method of iodoform 
injections is concerned, there is a certain difference, whether 
we dealt with internal abscess formation in the vicinity of the 
joint or simply with fungous increase of the synovial membrane. 
We evacuated all the tuberculous abscesses, as well as the joint 
abscesses, generally with a strong abdominal trochar in order to 
be able to remove the thick clots of cheesy tissue and the 
loosened pieces of the abscess membrane, sometimes contained 
in the pus. Through the canula we irrigated the abscess cavity 
with a three-per-cent. solution of boric acid until all the shreds 
of tissue were washed out and the water passed out clear ; then 



190 THE BACTERIOLOGICAL WORLD. 

we injected through the opening a ten-per*-cent. solution of 
iodoform (in pure glycerine or in water with some glycerine 
and mucilage of acacia ) into the abscess and the joint, sufficient 
to fill it slightly. 

The average quantity necessary amounts to 15 to 40 grams 
(about §ss-§ jss) ; I have occasionally gone higher, — in one case 
using 80 grams (gijss). I have never found it necessary to suture 
the opening produced by the large trochar ; it suffices if, after 
the withdrawal of the canula, the skin is drawn together into a 
fold, the little wound closed with a pad of gauze, and a slight 
compressing bandage placed thereon. 

When there appears in the abscess thick shreds of tissue, 
which continually stop up the canula, as does occur in the 
large burrowing abscesses, descending from the vertebral 
oolumn, the puncture should be enlarged to a sufficient length 
by introducing the knife upon the canula. This wound must, 
of course, be sewed before the use of the iodoform injection. 

For the treatment of tubercular joint diseases without 
discharge and suppuration, I used for puncture and injection a 
trochar which needs only to be 2 mm. ( T 1 ¥ in.) in diameter. It 
is much easier to penetrate the joint cavity with this than with 
the Pravaz needle. The quantity of the drug to be injected in 
such cases naturally corresponds to the smallness of the cavity ; 
.children often bear only 5 grams ( 5 jl )• 

In every case — with or without suppuration — while the 
opening is closed, one should, as much as possible, make 
passive motion in the operated joint, in a distributive way and 
in all directions ; by this means the entire joint region will be 
thoroughly massaged. In this way, I endeavor to obtain an 
equal distribution of the iodoform in all the folds and angles of 
the abscess and joint cavities ; perhaps on account of this, it 
happens that the iodoform is rubbed into the fungous masses, 
which are often so soft. 

The most convenient and proper place in which the indi- 
vidual joint should be opened by a trochar, I have determined 
by experimentation upon the dead body. The wristjoint is 
accessible on both sides, close under the styloid process of the 
radius and ulna ; the elbowjoint is reached by pressing deeply 
over the head of the radius, so easily perceptible in pronation 



THE BACTERIOLOGICAL WORLD. 191 

and supination. The shoulder joint, of which I have not had 
any disease to treat in this way, should be entered in a place 
external to the coracoid or external to the spine of the scapula 
in its passage to the acromion, where the swelling of the capsule 
of the joint and the abscess can be distinctly felt. The best 
for the injection into the hipjomt is to enter upon the 
greater trochanter with a trochar 7 to 9 cm. ( 2f to 3|- in.) long. 
In accordance with my autopsies, the following procedure is 
the most positive. While the patient is lying flat upon his 
back, the trochar is plunged perpendicular to the axis of the 
thigh immediately above the top of the trochanter major, 
midway between the anterior and the posterior edges, rather 
further in front, and is pushed slowly forwards until the sensa- 
tion of bone is perceived. On this account the leg must in no 
case be adducted and ever so little rotated outwardly, because 
the trochar might easily strike the margin of the acetab- 
ulum. Flexion, also, must of necessity be avoided with 
care. The best position for the thigh is adduction and 
slight internal rotation. If bone is felt with the trochar, 
it is the head of the femur, or the neck of the femur, 
near it. Thereupon the leg should be adducted as strongly as 
possible, and the trochar slipped in, somewhat above and more 
deeply, retaining throughout the bone sensation ( the head of 
the femur ) until bony resistance again interferes. Then it will 
be in the joint cleft between the femoral head and the margin 
of the acetabulum, and the stylet can be withdrawn, after which 
thecanula maybe shoved somewhat deeply into the joint cavity, 
and the injection given. A trocar 7 to 7.5 centimetres ( 2f to 
3 in.) long is the proper length for a fleshy adult ; in children, 
correspondingly less. 

This treatment permitted without difficulty the injection 
of 40 grams ( 5 X ) of the fluid into the hipjoint of a child, five 
years old, in which discharge was not apparent. Observation 
upon the dead body has certainly convinced me that in the 
living body the capsule of the hipjoint may be penetrated in 
the way described. In a case in which at first an abscess 
disappeared after the injection with iodoform, the fluid passed 
from the hipjoint into the old abscess cavity, an indication that 
a communication must still have existed between the hipjoint 



192 THE BACTERIOLOGICAL WORLD. 

and the old abscess. In two other cases the iodoform injection 
into the hipjoint made its appearance at fistulous openings, far 
from the joint, and on the anterior portion of the thigh. In a 
boy aged 13, the hipjoint was evacuated by a second puncture, 
and, while at first a discharge did not appear, about ateaspoonful 
of thick, stringy synovia came out of the canula, and in this 
case 60 grams ( 5 xv) was easily injected into the joint. 

The kneejomt is punctured in the usual way, viz., under 
the patella. In the anklejomt it is best to puncture perpen- 
dicularly under the apex of one of the malleoli, and then to 
move the trochar upwards. For the other joints no special 
directions are necessary. 

It has often happened to us, especially in dealing with the 
knee and the hipjoint, that, though the first injections were 
very mild, the later ones were given with difficulty and with 
much resistance. The reason is clear ; after the iodoform 
injections a change occurs in the diseased capsule which 
produces a cicatricial contraction and thereby a cure, at the 
same time, however, greatly diminishing the size of the joint 
capsule. 

It was very remarkable, in some cases, how very rapidly 
the general condition improved in the course of the treatment. 
We have very often observed this. A wonderful example is a 
case of kneejoint tuberculosis which I present to you in the 
person of this young man. His general condition had suffered 
uncommonly on account of the severity of his disease, and 
especially because of his sleepless nights ; just as soon, however, 
as the kneejoint disease improved, he developed strength, and 
now he enjoys blooming health. 

With respect to the remaining points, I must direct you 
to a communication which has already appeared in the Berliner 
Klinische Wochenschrift and to my forthcoming (in a few 
weeks) monograph upon i( Tuberculosis of the bone and joints." 



THE BACTERIOLOGICAL WORLD. 193 

THE FILTRATION OF WATER. 



BY W. E. DUCLAUX. 
(Translation from the French continued from February number.) 

All these bodies may, on the other hand, be affected by 
another means. They are more easily decomposable than the 
veritable albumenoid matters, under the action of the alkalies, 
which transform them into ammoniacal salts, as they do ordinarily 
all the amides. So that if we boil them with potash, and if 
we had in preference to this potash an oxidizing body as the 
hypermanganate is, which adds its destructive action on the 
organic matter, we will succeed in transforming into ammonia 
the total or nearly the total of the nitrogen of the amides, 
whilst on the contrary the complex albumenoid matters would 
be more resistent. 

This ammonia resulting chiefly from the transformation 
of the amides should be, in its turn, distinguished from 
ammonia which may exist already formed in water. All- 
waters contain a little, and, when there is a little, the significance 
of the presence of this body is hardly clear. Some of it is 
produced by the action of microbes on nitrogenized matters,, 
and consequently its presence may be evidence of the existence 
in water of the products of fermentation of animal matters, 
perhaps also of the products of the decomposition of urea,, 
which a diastase transforms easily into carbonate of ammonia. 
But, on the other hand, this ammonia is formed principally, 
as I have shown, during the life of aerobic microbes, which 
of all germs are evidently the least dreadful, and as there are 
microbes everywhere, and we cannot avoid them, it is better to 
have to deal with the most harmless. Finally, this ammonia 
of water may serve in its turn for other microbes, and by their 
assistance reach the rank of albumenoid matter. 

All these opposite actions render very uncertain the diag- 
nostic value and the significance of the presence of ammonia in 
water. Still we should consider it as decidedly unfavorable 
when its proportion reaches or exceeds one or two milligrams 
per litre, and it is better that water contain none. 

This ammonia can be measured with great precision in 
distilling a half litre of water with a few drops of a potash 



194 



THE BACTERIOLOGICAL WORLD. 



solution, and in treating by means of ten-per-cent. solution of 
sulphuric acid or oxalic acid the first portions of the liquid 
distilled. When this operation is ended, and before the 
remainder of the liquid is cooled, we add to it 20cc. of a 
solution containing 8gr.of hypermanganate of potash and 200gr. 
of caustic potash per litre. We begin to distil over again ; we 
take 50cc. two or three times, and of each of them we measure 
the ammonia. When the last contains no more, we stop, we 
add the total, and we have the ammonia coming chiefly from 
the hydration of the amides. It is this ammonia that we record 
under the singular term of albumenoid ammonia. It is certainly 
the albumenoid matters which furnish the least of it, and it is 
chiefly an evidence of the presence in water of the products 
of the most advanced destruction of these albumenoids ; it is 
even this that gives it its principal interest. 

We may now form an idea of the transformation brought 
about in the water which filters, by the action of the microbes 
which it meets, especially in the upper layers of the sand. 
We may consult the excellent publications of the commission 
that the waters of the city of Zurich, where the oxidability, 
albumenoid ammonia, the free ammonia and the number of 
bacteria per litre are simultaneously determined. Here is in 
milligrams the average figures for 1888. We have replaced the 
figures relative to the oxidability by the number of the 
milligrams of oxygen yielded by the hypermanganate and 
the organic matter of one litre of water. 





Consumed 
oxygen. 

3.76 

3.04 


Albumenoid 
ammonia. 


Free Am- 
monia. 


Bacteria. 




0.039 
0.023 


0.009 
0.003 


188,100 
1,190,000 







Consulting these figures, which resemble those of preceding 
years, we see that the water loses in passing through the filter 
the greatest portion of its free ammonia, a portion of its organic 
matter, and as much as we can judge by these fatally incorrect 
figures, the more notable fraction of its nitrogenized elements 
which are close to the group of amides. Conclusions are the 
same concerning the waters of Berlin. 



THE BACTERIOLOGICAL WORLD. 



195 



Where is the principal seat of this destruction of organic 
matter ? An interesting experiment of Mr. Piefke shows that 
it is principally in the superior layers, the richest in microbes. 
Through three filters of the same sand, and the thickness of 
which, from 0m7, lm4 and 2ml, were to one another as the 
numbers 1, 2 and 3, it filtered the same water with the same 
speed, analyzed at its entrance, and at the exit from a point of 
view of oxidability and concerning the quantity of free oxygen 
contained. Here are the relative figures on the thirteenth of 
January, 1889. I give, as comparison, the result of the indus- 
trial filtration of this water from a microbe point of view, the 
same day. The oxidability is estimated as above in milligrams 
of oxygen, the free oxygen in cubic centimetres per litre : 



BEFORE FILTRATION. 



Oxidability . . 
Free oxygen 
Bacteria 



6.6 
7.9 



Filter of 0m7, 



5.1 

4.8 



62,840,000 



AFTER FILTRATION. 



Filter of lm4. 



4.7 
3.1 



Filter of 2rnl. 



3.700,000 



4.6 
4.6 



If we would have an idea of what transpires in the 
first third and in the two first thirds of the thickest 
filter, we have evidently only to subtract one from the 
other the figures relative to the oxidability and the free oxy- 
gen borrowed from the three filters which have served for 
the experiment. We have then, in decomposing in our 
mind tbe filter of 2ml into three superposed filters of 0m7 of 
thickness, lmg5 for the variation of the oxidability of the 
superior filter, 0mg4 for the middle filter, Omgl for the inferior 
filter. The effect produced decreases then in the same way 
that the number of bacteria does from the top to the bottom 
of the filter. Undoubtedly, this coincidence does not suffice to 
establish their relation from cause to effect ; we might say 
again that, if combustion is not carried on equally through the 
layers of the filter, it is because the oxygen is scarcer and 
scarcer as we descend, as is proven by the very figures of the 
preceding table. We may answer that after the passage 
through the filter of lm4, there remained still 3cc. of free 
oxygen from 8 which existed at the beginning. Besides in a 



196 THE BACTERIOLOGICAL WORLD. 

sterilized filter in which the water arrives drop by drop so 
that it remains penetrated by oxygen, there is no percepti- 
ble combustion. We must, therefore, forsake the idea of seeing 
in this combustion a phenomenon of ordinary chemical oxida- 
tion. The same experiments cited above with the sterilized filter 
precludes the attribution of the diminution of the organic 
matter to a wall action in passing through the filter. There is- 
nothing left, then, in it, than the result of the vital action met 
in all the thickness of the filter but chiefly in the superior 
layers. 

If it is thus, we get a reply to the question that we had 
asked ourselves above, without being able to solve it ; it is 
evidently necessary that water remain a certain time in the 
filter to allow the microbes to exert their influence. We can 
ascertain this duration of contact either in diminishing the pres- 
sure on the water, or in augmenting the thickness of the filter 
for the same pressure. The first method is evidently the most 
economical, and experience has taught in each case what combi- 
nation between the filter and the pressure was the best, to bring 
about the most notable increase in the number of bacteria and 
in the quantity of organic matter in water after filtration. As 
we may foresee, this combination varies with the degree of the 
impurity of the water. Thus at Berlin, at the factory of Stra- 
lauer Thor, where water of the .Spree is used, which is in general 
impure enough, a rapidity of lml8 per day is scarcely exceeded. 
This rapidity may reach three metres in the establishment 
where water of Tegel lake is filtered which is generally much 
less laden with germs and organic matters. Finally we have 
seen that at Zurich, where the waters of the lake are still 
purer, twenty-eight metres per day could be passed through the 
filter without compromising its working perceptibly. 

We see clearly now what is a sand filter. The sand serves 
both as a break to moderate the movement of the water, and as 
support, for the slimy coating of microbes which forms itself in 
all its thickness, but principally on the surface. This superficial 
coating becomes, when it is formed, the true filtering layer, and 
after having operated mediumly until then the filter is at last 
ripe and complete ; but this filtering coating is a fragile thing. 
We must not submit it to too strong pressure when it is weak ; 



THE BACTERIOLOGICAL WORLD. 197 

its elements would break loose, would be carried away in depth 
of a filter and obstruct it. We must not either submit it to 
rapid variation of pressure which would produce the same 
effects. We must let it work slowly, increase the pressure 
gradually in proportion to the increased thickness of the 
coating, and according to its increasing resistance and imper- 
meability ; then, at a given moment, when the pressure used 
has become too great stop the water, let the filter exhaust itself, 
remove the superior filthy coating and put the filter in operation 
again. The space of time between two cleanings is called a 
period. It is evidently the shorter — everything being otherwise 
equal, that the water to be filtered is dirtier and more impure. 
It is thus that at Berlin, at Stralauer Thor, the average dura- 
tion of the period in 1888 has been sixteen days with an 
average speed of 1ml per day, whilst the Zurich, has been in 
1887, for a covered filter forty days, with an average speed of 
4m5 per day. — Annates de L'Institut Pasteur. 

[To be continued.'] 



EHRLICH'S TEST IN TYPHOID FEVER. 

By Charles E, Simon, M. D., Assistant Physician, Johns Hopkins 

Hospital. 

A Paper read before the Johns Hopkins Hospital Medical Society, 

October 20, 1890. 

It had long been known that when amido derivatives of 
the hydrocarbons of the benzine series are treated with nitrous 
acid, compounds are obtained which, owing to their extreme 
instability, and the readiness with which they enter into combi- 
nation with an almost infinite number of other bodies, espe- 
cially with aromatic compounds, have become of great practical 
as well as theoretical importance. 

Particularly interesting is the fact that a very large number 
of our dyes are derived from these bodies. They were discov- 
ered by Peter G-riess, and termed by him diazo-compounds. 

With the expectation of demonstrating by a color reaction 
some of the aromatic bodies, the existence of which in the 
urine had been rendered highly probable by the researches of 



198 THE BACTERIOLOGICAL WORLD. 

Baumann and Brieger especially (Zeitschrift f. Physiol. Chemie, 
Bd. 3), Ehrlich undertook a series of carefully conducted 
experiments in this direction. 

A preliminary account of these investigations he published 
in the Zeitschrift f. Klin. Med., Bd. 5, in 1882, which was 
followed by a more complete account of his results in the 
Gharite Annalen, 8ter Jahrgang, in 1883. 

The practical value of these, especially as regards their 
diagnostic importance in typhoid fever, has been the subject of 
much controversy, and it appears at the present day that the 
so-called Ehrlich 's reaction is regarded by many as a medical 
curiosity of absolutely no diagnostic importance whatsoever ; 
Petri and Penzoldt are the exponents of this side ( Penzoldt, 
Berl.Klin. Wochenschrift, 1883 and Petri, Zeitschrift far Klin. 
Med., 1884). The manner, however, in which these two 
observers entirely disregard the instructions given by Ehrlich, 
in order to insure success in the simple experiment, deprives 
their work of all value ; for, as Ehrlich says, " there are but few 
tests in chemistry which cannot be spoiled by careless manip- 
ulation." (Ehrlich, Deutsche Med. Wochenschrift, 1883.) 

A great deal of confusion has also undoubtedly arisen from 
the fact that 1 the exact color, produced in the experiment, has 
either been described improperly, or simply spoken of as 
the " characteristic reaction," a term conveying no idea what- 
soever. As the original work done by. Ehrlich does not seem 
to be fully appreciated, it is my purpose here to describe the 
methods of testing which I have found most useful, and show 
that my results, on the whole, coincide with those which he 
obtained. 

Since the preparation of chemically pure, crystalline diazo- 
compounds is quite a difficult process, Ehrlich made use of the 
fact that sulphanilic acid, when treated with nitrous acid in a 
nascent state, forms in solution the diazo-benzine-sulphonic 
acid, which thus becomes the active principle in the mixture 
employed. 

The sulphanilic acid, used for this purpose, is the para- 
amido-benzine-sulphonic acid, prepared on a large scale, and 
used extensively in the coal-tar color industry. 



THE BACTERIOLOGICAL WORLD. 199 

Other compounds, of course, can also be used, as the meta- 
amido-benzine-sulphonic acid, the ortho and paratoluidine- 
sulphonic acids, and others, but among all these Ehrlich 
has found the common sulphanilic acid the most convenient. 

Two solutions are employed and kept in separate bottles, 
the one containing 50 cc. of hydrochloric acid, which is diluted 
to 1,000 cc. and saturated with sulphanilic acid, the other being 
a ^ per cent, solution of sodium nitrite. 

To make the test, 40 cc. of the sulphanilic acid solution 
are taken in a measuring glass, and 1 cc. of the sodium nitrite 
solution added, the mixture being thoroughly agitated. The 
hydrochloric acid now acts upon the sodium nitrite, forming 
nitrous acid which, in a nascent state, forms the diazo-benzine- 
sulphonic acid by its action upon the sulphanilic acid. Small 
quantities of the sodium nitrite are used, and the absence of 
any free nitrous acid in the mixture is thus insured, and at the 
same time very small quantities of the diazo-benzine-sulphonic 
acid are formed — one of the principal requirements to insure 
success in the experiment. 

The reaction which thus takes place is represented as 
follows : 

I. NaNO 2 + HCl=Na CI -f- HNO 2 

/NH 2 /"N\\ 

II. C 6 H 4 -f HN0 2 =C 6 H 4 N+2H 3 0. 

\S0 3 H \S0 3 / 

Para-amido-benzine-sulphonic acid. Diazo-benzme-sul- 
phonic acid. 

In his original article, Ehrlich advised the addition of this 
mixture to the urine, to be tested, in the proportion of 1 : 1 
per volume. If ammonia is added in excess to the urine thus 
treated, the color play, presently to bs described, occurs. In 
a later communication (Charlie Annalen, Bd. 11, 1886), he 
has modified this method by mixing one volume of urine with 
5 to 6 volumes of absolute alcohol, previous to the addition of 
the sulphanilic acid mixture, filtering, and then adding the 
acid mixture to the filtrate. 

I have found it convenient to add about 50 cc. of absolute 
alcohol to 10 cc. of urine, filtering, and then running into the 
alcoholic urine, which has become more or less decolorized, the 



200 THE BACTERIOLOGICAL WORLD. 

sulphanilic acid mixture from a burette ; 20 cc. of the latter 
are then sufficient, added to about 30 cc. of the alcoholic urine ; 
the addition of the acid in small quantities, for example, 2 cc. 
at a time, followed by thorough shaking of the urine, is at 
times advantageous, especially in typhoid fever, when the 
disease has advanced to a point at which the color reaction is 
no longer at its original intensity. By the addition of a few 
drops of ammonia to the final mixture, the characteristic color 
appears in typhoidal urine to disappear on shaking, and to 
become permanent after an excess of the ammonia has been 
added . 

I have found a small Erlenmeyer's flask more convenient 
for holding urine than the ordinary test-tube, the exact shade 
of color being more apparent by transmitted light. With this 
modified method most of my experiments have been performed. 
There is, however, a third, which I consider more convenient, 
less expensive, and more delicate. A few cc. of urine are taken 
in a small test-tube, and an equal quantity of the sulphanilic 
acid mixture added, the whole being thoroughly agitated ; 1 
cc. of ammonia is then allowed to carefully run down the side 
of the tube, forming a colorless zone above the yellow urine, 
containing the acid, and, at the junction of the two, a more or 
less deeply colored ring will be seen, the color of which is 
readily distinguished and noted, the slightest carmine tint 
being made out more readily by its contrast with the colorless 
.zone above, and the yellow below, than when we are dealing 
with a uniform color. 

This simple modification I recommend particularly, and as 
it occupies but a few minutes, I am sure it will be found 
especially convenient by the busy practitioner. As to the color- 
play which occurs in different kinds of urine, it will be observed 
that in normal, or pathological, but non-febrile urines, the 
color of the pure, or alcoholic urines, when method No. 2 is 
employed, remains either unaffected or is merely intensified by 
the addition of the ammonia ; a deep orange tint may even be 
produced in this way, but is of no significance whatsoever, and 
easily enough distinguished from the typical color. Ehrlich 
records one exception to this general rule, namely, that in 
urines containing biliary coloring matters, an intensely dark, 



THE BACTERIOLOGICAL WORLD. 201 

-cloudy discoloration occurs at times which, upon boiling, is 
changed to an intense reddish-violet color. 

In the course of my experiments I have met with "another 
very interesting exception, but regret that I have but one 
observation to record on the case, which I owe to the kindness 
of Dr. Ogden of Milwaukee. The urine of this case contained 
a substance which reduced Fehling's solution, but did not 
reduce the subnitrate of bismuth, and merely produced a black 
discoloration, and in which the fermentation test failed com- 
pletely. Undoubtedly this was one of the rare instances in 
which glycuronic acid, first isolated by Schmiedeberg and 
Mayer (Zeitsclirift f. Physiolog. Chemie, Bd. 3, 1879), occurred 
normally in the urine. When Ehrlich's test was applied to 
this urine, according to the second method, described above, a 
dark brown color developed on standing for a quarter of an 
hour, which, at the end of an hour, became almost black. As 
regards febrile urines, Ehrlich observed the production of an 
intensely yolk-yellow color, which was even imparted to the 
foam when method No. 1 is employed, in rare instances of 
endocarditis ulcerosa, abscessus hepatis, and intermittens, 
especially, i. e., in diseases associated with well-marked 
chills. 

'Now, in typhoid fever, and this, of course, is the most 
important, a color occurs upon addition of ammonia, which 
may vary from an eosin to a deep garnet. Here I found method 
No. 2, but particularly No. 3, very useful, because with them 
the production of the faintest rose-tint is more readily per- 
ceived than when No. 1 is employed, owing to the fact that in 
the second method we are practically dealing with a primarily 
colorless solution, and in No. 3, as above stated, we can take 
advantage of the contrasts. 

If method No. 1 is used, and there be the least doubt as to 
the presence of the characteristic color, which may occur in the 
later stages of typhoid fever, the reaction may be said to be 
present when the foam which appears on shaking shows a 
reddish color, and especially when upon standing for twelve to 
twenty-four hours the superficial layer of the sediment shows a 
green color. This latter is found to be due to the transforma- 
tion of the red soluble coloring matter into an insoluble green 



202 THE BACTERIOLOGICAL WORLD. 

body. When the color is of its maximum intensity, of course- 
no difficulty is experienced whatsoever in recognizing the 
presence, of the characteristic reaction, the color being then of 
the most intense carmine or garnet shade. 

That this reaction is not dependent upon the presence of 
any substance, normally present in the urine, that Ehrlich 
seems to have established beyond a doubt, but as to the nature 
of the substance producing the reaction we have absolutely no 
knowledge. It is possible that a careful chemical study of the 
green sediment will lead to a clue as to the nature of this body, 
but this, of course, must be left to the hands of chemists. 

Ehrlich has tested the urines of patients, afflicted with 
almost every known disease, and has arrived at negative results, 
with some exceptions ; typhoid fever, of course, being the most 
important and interesting. At times, however, he has met with 
the same in some of the morbilli, and diseases in general char- 
acterized by high fever, as in severe cases of phthisis pulmo- 
nalis, although it may occur in the latter in the absence of 
fever, but when present for some time he regards it a bad 
omen. 

Although I have tested the urine of almost every disease 
which has occurred in the medical wards of our hospital, 
besides a number of specimens taken from healthy individuals, 

I have only observed it in cases of typhoid fever and phthisis 
pulmonalis. Not having as yet had opportunity of examining 
specimens taken from contagious cases, I am unable to express 
any opinion on these. On phthisis pulmonalis I wish to report 
at some future occasion. I subjoin a summary of the typhoid 
fever cases in which Ehrlich/s test has been used. The degree 
of reaction is noted as follows : III signifies very intense ; 

II intense ; I marked ; l)i faint. 

Case 1. — George N., aet. 35, admitted July 18, the fifth 
day of disease. Reaction on the sixth, seventh and eighth 
days. Patient left the hospital before recovery. 

Case 2. — Charles B. S,, aet. 51, admitted July 2, the 
fifteenth day of the disease. Death resulted on the twenty- 
fourth day of the disease. The reaction was very faint on the 
sixteenth to eighteenth day, and absent on the nineteenth day. 



THE BACTERIOLOGICAL WORLD. 



203 



Case 3. — Thomas K., aet. 21, admitted July 8. His 
primary illness lasted 20 days,, and succeeding a period of 7 
afebrile days, a relapse occurred lasting 11 days, the case end- 
ing in recovery. T=100°-104: F. 

On the 7th day of the disease the reaction was III. 



it 


8th 


i t 




a a 




III. 


it 


9th 


i t 




i t a 




II. 


(I 


10th 


a 




it a 




I. 


it 


11th 


a 




a a 




Iji. 


a 


12th 


'it 




a a 




0. 


Meth 


od No. 2 


was used. 


Not tested 


in 


the relapse. 



( a 


III. 


i a 


III. 


i a 


III. 


i a 


II. 


i a 


I. 



Case 4. — Frederick T., admitted July 8. His primary 
illness lasted 17 days, and succeeding a period of 13 afebrile 
days, a relapse occurred, lasting 11 days, ending in recovery. 
T=100.2°-103.5°. 

On the 6th day of the disease the reaction was III. 

7th 

8th 

9th 

" 10th 

" 11th 

Method 2 used. Not tested during the relapse. 
Case 5. — Charles E., admitted July 16, with a possible 
relapse, the. fever continuing uninterruptedly for 55 days, and 
ending in recovery. T=100°-104.5°. On sixteenth day of 
disease reaction was 0. During relapse present for several days 
III. Method 2 used. 

Case 6. — John M., admitted August 21, the fever lasting 
35 days, ending in recovery. On fourteenth day of disease 
reaction was II. 

Case 7, — Joseph D., admitted August 21. Death resulted 
on the nineteenth day. T=100°-104 c> . 

Eeaction on 10th day III. 
(i 11th " II. 
" 12th " I/a. 
Case 8. — Mary S., aet. 11, admitted September 9. Her 
primary illness lasted 27 days, and was followed by a relapse 
continuing for 12 days, ending in recovery. Method 2 used. 
Reaction absent throughout. This case and the following, 



204 



THE BACTERIOLOGICAL WORLD. 



also, occurring in a child, are my only ones in which the reac- 
tion, which was expected a priori, was negative. Both cases 
were of considerable severity, the fever remaining between 103° 
and 105° for some days. 

Case 9. — Lizzie X., aet. 12, treated outside the hospital, 
the illness lasting 23 days. Eeaction was not obtained after 
the eighth day. 

Case 10. — Francis H., aet. 10, admitted September 28. 
Temperature gradually fell from 104° and ended in recovery. 
On the 7th day of the disease the reaction was III. 



8th 


a < 


i a 


III. 


9th 


ti 


'.( k 


III. 


" 10th 


t( 


it tf 


III. 


" 11th 


a 


it 


II. 


" 12th 


i ( 


at a 


I. 


" 13th 


ft 


' ( . i 


0. 



Case 11. — John B., aet. 19, admitted September 27. 

T=99.5°-102°, reached 104° once. Eecovery. 

On the 17th (?) day of disease the reaction was II. 

18th " " " " I. 

<< 19th (i « << « 0# 

Case 12. — Anton D., aet. 25, admitted September 25. 
T=100°-105°. Reaction on the thirteenth day Ija. 

Case 13. — Albert S., aet. 55, admitted September 23. 
On twenty-second day of illness reaction was III ; on the four- 
teenth day the temperature had reached 97.5°-99.2°, probably 
indicating a pseudocrisis. 

Case 14. — James McD., aet. 27, admitted October 15. A 
critical case, temperature 103°-105o, reduced by baths. Ring 
method used. Reaction present from fifteenth to nineteenth 
day II. 

Case 15. — Thos. O'D., aet. 23, admitted September 20. 
T=100°-104°, often reaching 104°. Reaction on fifteenth 
day I. 

Case 16. — John F., aet. 20, admitted October 9. Pseu- 
docrisis on nineteenth day ; temperature 100°-104°. On six- 
teenth day no reaction with method 2. With ring method 
present II from twenty-first to twenty-fourth day. 



THE BACTERIOLOGICAL WORLD. 



205 



Case 17. — Samuel H., aet. 21, admitted October 9. Very 
mild case ; temperature normal from eleventh day. Splenic 
enlargement still present on sixteenth day with normal temper- 
ature. ~No reaction was obtained. 

Case 18. — Timothy M., aet. 24, admitted October 9. 
T=99°-103°. Reaction present on the seventh and eighth 
days III ; and on the ninth day II. 

Case 19. —Otto K., aet. 22, admitted October 6. T=100°- 
103°. On tenth day reaction I. . Ring method, on sixteenth 
day, I//. 

Case 20. — James D., aet. 18, admitted October 10. 
T=99.5°-104°. No reaction on the twenty-second day. 
Case 21. — Charles B., aet. 27, admitted October 15. 
Ring method on 18th day was II. 
" 19th " II. 
" " " 20th " I. 

" 21st " I, 

" 22d " I. 

Case 22. — Robert J., aet. 24, admitted October 7. 
T=100°-102°. Mild case. Reaction on sixteenth day. 

Case 23. — Annie S,, aet. 28, admitted October 13. Crit- 
ical case. T=100°-104.5°. 

Reaction with ring method III on 5th day. 

Ill " 6th 
III " 7th 
III " 8th 
III « 9th 
II " 10th 

Case 24, — Maggie B., aet. 19, admitted October 13. 
T=100°-105°. Critical case. Reaction with ring method 
present from fourteenth to twenty-first day III— II. 

Case 25. — Clara W., aet. 24, admitted October 28. 
T=100°-104. 

Method 2 ; reaction present on 5th day III. 



a 


a 


a 


a 


t t 


a 


(( 


a 


a 


a 


a 


a 


it 


a 


a 


a 


a 


a 


a 


a 



a 


a 


" 6th 


a 


III. 


a 


a 


" 7th 


a 


III. 


a 


a 


« 8th 


a 


I. 


a 


a 


" 9th 


a 


I. 



206 THE BACTERIOLOGICAL WORLD. 

Case 26. — Ed. Y., aet. IB, admitted October 14. 
T=101°-104°. 

Eeaction present on 7th day III. 
" " (i 8th " III. 

From these cases it will be seen that the reaction was only 
absent in four cases, of which two were the children above 
mentioned, one a very mild case, lasting but eleven days, and 
one tested on the sixteenth day of the disease, at a time when 
the reaction is not always obtained and, in fact, is usually 
absent, 

CONCLUSIONS. 

First. The reaction may be obtained in cases of typhoid 
fever from the fifth to thirteenth day without any difficulty 
with methods 1 and 2. 

Second. With method 3 it may be observed as late as 
the twenty-second day, at a time when neither 1 nor 2 will 
yield absolute results. 

Third. Its absence from fifth to ninth day indicates a 
very mild case, excepting in children, although this rule is 
probably not an absolute one. 

Fourth. As it occurs previous to the appearance of the 
rash, it is a very useful aid in the diagnosis in typhoid fever. 
— The Johns Hopkins Hos. Bulletin. 



THE CENTENARY OF THE MICROSCOPE. 



BY ERNESTO MANCINI. 



Nuova Antologia, Rome, December. 

It is generally thought that the invention of the microscope 
goes back to the close of the fifteenth century, or, to be more 
precise, to the year 1590, when, in the city of Middleburg, in 
Holland, two spectacle-makers, named Janssen, invented both 
the telescopy and the microscope. This date for the invention, 
according to which its third centenary would arrive in 1890, 
does not rest on authentic documents, but is based on assertions 
published in 1665 by the physician, Peter Borel. He denied 



THE BACTERIOLOGICAL WORLD. 207 

-that Galileo, Drebbel, and others, deserved the credit of having 
invented the telescope ; and in order to demonstrate that the 
invention of that instrument, as well as of the microscope, was 
due to the Janssens, produced some documents which showed 
that the two spectacle-makers, having invented the telescope in 
1590, presented a specimen of it to Prince Maurice, Stadtholder 
of the Netherlands, and to the Archduke Albert. Later on, 
however, the telescope of Prince Maurice became a microscope, 
in a letter of William Borelli, who declared that he had always 
heard in Middleburg, his native city, that the Janssens had 
invented these optical instruments, and further that, when he 
was ambassador at London, in 1619, he had seen in the hands 
of Drebbel the identical microscope that the Janssens had pre- 
sented to Prince Maurice. 

Professor Govi, however, in a work which demonstrates the 
excellence of his judgment and his vast erudition, has collected 
a series of documents, which not only seem to restore the merit 
of the invention of the microscope to Galileo, but show the 
various vicissitudes of the discovery itself. The first hint of 
the transformation of the Holland telescope into a microscope 
is found in a little book published in 1610 by WodderbOrn, a 
pupil of Galileo. Speaking of the wonderful qualities of the 
telescope, Wodderborn adds, in praise of Galileo, that " with 
the instrument could be perfectly distinguished the organs of 
motion and sensibility in the smallest animalcule/'' so that the 
particular formation of multiplied eyes in very small animals 
could be perceived. This new application of the telescope by 
himself Galileo did not deny, though he never directly affirmed 
it. In the National Library at Paris is preserved a letter by 
Canon Tarde, in which he speaks of visiting Galileo in Florence, 
•in 1614, when the latter was sick in bed. Notwithstanding, to 
Tarde Galileo gave ample explanation of a microscope then in 
his possession. 

Whether the invention of the simple microscope be due to 
Janssen or Galileo, to Drebbel is due the merit of having pro- 
duced at Rome, in 1624, the compound microscope. The 
difference between the two hardly needs explanation. The 
simple microscope magnifies with a single lens, or with several 
lenses so close together that they act like a single lens. The 



208 THE BACTERIOLOGICAL WORLD. 

compound microscope has two or more lenses, separated by a- 
convenient distance from each other, and which act separately. 
In 1669, Eustachio Divini constructed a colossal microscope 
which magnified 140 times. A little after, Bonannus invented 
a horizontal microscope which magnified 300 times. 

In the seventeenth century were laid the foundations of 
micrography, a science which, by the study of the minute 
anatomical elements and their functions, has made such great 
progress under the name of histology, and been such a fertile 
cause of important discoveries. With the microscope, Mal- 
pighi, by the minute examination of the tissues, confirmed the 
theories about them he had previously formed ; Leuwenhoek 
discovered the globules of the blood and the structure of the 
nervous fibres ; and Swammerdam dissected insects, of the most 
minute organs of which he gave descriptions still considered 
perfect. 

In the eighteenth century, observes Henocque, but few 
modifications were made in the microscope. To mention all 
the improvements made in the instrument during our century 
would be tedious. During the last forty years enormous 
advances have been made in science by the aid of the micro- 
scope, of which the usefulness has been greatly increased by the 
skill with which the matter to be examined is prepared, and 
by the aid of photography. Microphotography dates from 1840 
only ; but since that date it has had an uninterrupted series of 
noteworthy improvements. 

Besides histology, created by the microscope, by which, 
our acquaintance with the most hidden structure of organisms is 
constantly increasing, bacteriology, with its rapid succession of 
discoveries of the highest importance, owes its existence to the 
microscope. Those little beings, those micro-organisms, which, 
by the change of the medium in which their evolution is 
effected, can produce so much good or so much evil, and of 
which it takes several millions to occupy the tenth part of half 
an inch in space, can now be identified according to their 
species, notwithstanding their changeable aspect. We can 
estimate the rapidity of multiplication, the number, the dimen- 
sions, and the singular manner in which by dividing themselves, 



THE BACTERIOLOGICAL WORLD. 209 

or by means of a sort of buds or spores, the micro-organisms 
reproduce themselves. 

In the examination of the inorganic world the microscope 
has had results not less precious. The wonderful phenomena 
of crystallization, the exact form of the crystals, the more pre- 
cise in proportion to their minuteness, the modifying proper- 
ties of the light called forth by the thinnest layer of a mineral, 
the interior texture of rocks, v all these can be studied with a 
precision impossible before the invention of the improved 
microscope. And, finally, not to mention all the triumphs 
achieved by the instrument, it has had an application which 
formerly would have seemed paradoxical, since the microscope 
has been employed to show the particulars of the nature of the 
surface of the planets, particulars which have been made clear 
by microscopic observations of instantaneous photographs. — 
Literary Digest. 



SCIENTIFIC GLEANINGS. 



Effect of thunderstorms on milk. — The effect of 
thunderstorms in turning milk sour is a matter of con- 
stant observation in every household. It is not certainly 
known to what element in the air this souring action 
on milk is to be directly attributed, and most people are content 
to ascribe it to " electricity in the air." An Italian savant, 
Professor Gr. Tolomei, has lately made some experiments with 
the view of elucidating this question. He found that the 
passage of an electric current directly through the milk not 
only did not hasten, but actually delayed, acidulation, — milk 
so treated not becoming sour until from the sixth to the ninth 
day, whereas milk not so electrified became markedly acid on 
the third day. When, however, the surface of a quantity of 
milk was brought close under the two balls of a Holtz machine 
the milk soon became sour, and this effect he attributed to the 
ozone generated ; for when the discharge was silent the milk 
soured with greater rapidity than when the discharge was 
explosive, — in the former case more ozone being formed than in 



210 THE BACTERIOLOGICAL WORLD. 

the latter. The souring of milk is generally attributed to the 
growth of a ferment ( bacterium ), which converts the milk 
sugar into lactic acid. It is possible, then, that the presence 
of ozone in the air overlying the milk hastens the growth 
and multiplication of the bacterium. The first observation, 
namely, the retardation of souring by the passage of a current 
through the milk, may be a point of practical importance to 
milk traders. Kegarding this theory that milk becomes sour 
from the action of ozone, Dr. Henry McOlure ( British Med- 
ical Journal ) contends that there must be some mistake. He 
writes : "One can hardly imagine that this gas, which undoubt- 
edly possesses powerful oxidizing and antiseptic qualities, should 
be the factor in producing this result. The result may be 
.admitted, but viewed in relation to the experiments of Professor 
Lodge, confirmed by myself, on the precipitation of smoke by 
the static spark, and stated in my book on ' Static Electricity 
in Medicine/ it is a fair inference that the atmospheric dust, 
together with organic germs, are thus precipitated during a 
thunderstorm, the precipitation of the latter causing the rapid 
souring of milk, the putrefaction of meat, etc.; and thus there 
seems a scientific basis for the popular belief that electrical 
storms clear the air." — Druggists Circular. 

SCHOLL ON INVESTIGATIONS INTO CHOLERATOXINES. — 

The conclusions of the author's experiments, made in the 
Hygienic Institution of the ( German ) University of Prague, 
are as follows : " The toxic peptone which I have produced 
from genuine albumen, after its destruction by anaerobiosis, is 
quite distinct from Petri's toxo-peptone (formed by aerobiosis ) 
because the latter is not destroyed by boiling, while mine is 
rendered inert. My choleraicpepto-toxine shows far more 
poisonous and characteristic properties than the toxines of 
Brieger and Petri formed by aerobiosis in peptone solutions, 
for the toxine which I procured ( by the cultivation of cholera 
bacilli ) from a single egg was sufficient to kill ten guinea-pigs 
in ten minutes with acute paralytic symptoms. The assertion 
of Hueppe and Wood, that more and stronger toxines are 
formed by cholera bacilli by anaerobiosis than by aerobiosis, is 
fully confirmed by my experiments, which negative the opposite 
assertions of Petri." — London Medical Recorder. 



the bacteriological world. 211 

Behavior of the virus of cholera, enteric fever, 
and of tuberculosis in" milk, butter, whey and cheese. 
Milk having been shown to be a vehicle for the transmission of 
certain diseases, e. g., scarlet fever, enteric fever, cholera and 
tuberculosis, Dr. L, Heim made experiments to ascertain the 
duration of viability of certain disease germs when cultivated 
in milk or the food-stuffs prepared from it, butter, whey and 
cheese. The author's results show that the germs were still 
capable of development in 

Cholera. Enteric Fever. Tuberculosis. 

Milk after 6 35 10 days. 

Butter " 32 21 30 " 

Curds " 1 2 " 

Whey 2 1 14 " 

Cheese 1 3 14 " 

— Journal Royal Microscopical Society. 

Peroxide of hydrogen as a sterilizer of milk. — 
Dr. Heidenhain ( Centralblatt f. Bakt. u. Pafasitenkunde ), 
referring to Dr. Altehoefer's researches on the use of peroxide 
of hydrogen in the disinfection of water, says that it may also 
be useful for sterilizing milk, although he is not able to say as 
yet how the digestibility of milk so treated will be affected. He 
finds that children take it readily, and that the peroxide, 
mixed in the proportion of five or six tablespoonfuls to one 
litre of milk, does not cause the milk to curdle or to become 
sour, even when it is left in a room at summer temperature for 
forty-eight hours, the slight acid reaction with litmus paper 
being due to the action of the disinfecting material and not to 
the prodaction of lactic acid. The cream that rises is so sweet 
that butter cannot be made from it for a very considerable length 
of time. — Druggists Circular. 

Pathogenesis of tetanus. — Faber (Berliner Klin. 
Wochenschr.) has separated by filtration, from pure cultures of 
virulent tetanus bacilli, a substance that is wholly free from 
germs, and yet when it is injected into animals, either in the 
cellular tissue or blood, it produces the clinical picture of 
tetanus. There seems to be a certain latent period, before the 
production of the symptoms, that varies with the virulence of 



212 THE BACTERIOLOGICAL WOULD. 

the culture from which the extract is prepared. When injected 
into the blood it produces general convulsions ; when into the 
cellular tissue these are associated with local convulsions. No 
specific immunity could be determined as a result of the injec- 
tions. — Jour. Am. Med. Ass'n. 



TECHNIQ UE. 



Introduction of a universal scale of magnification 
of microscopical figures.* — M. P. F. Reinsch complains 
that every micrographer makes use of an arbitrary scale for his 
figures. The camera obscura gives, for the same combination 
of objective and eye-piece, a constant magnification of the 
object figured ; but the microscopes of different makers differ 
widely, and the magnifications of the systems of objectives and 
eye-pieces do not vary in the same proportion. Consequently, 
the statement as to the eye-piece and objective employed, made 
by authors on their figures, conveys little meaning to those who 
have not the same microscope at their disposal. 

In the systematic description of microscopical plants, it is 
of the utmost importance to give the dimensions in absolute 
value. For microscopical work such as this, the micron /x has 
been chosen as the unit of measure-. A scale of magnifica- 
tion should be also based on the same unit. Measurements are 
indispensable in order to compare specimens of algae and 
microscopic fungi witli the published figures, but the work is 
complicated by the variety of magnifications used by authors, 
and long calculations are often necessary in order to find the 
absolute numerical values required. . 

To draw figures in conformity with the base of the 
measurements, the microscope must be brought to the desired 
magnification. 

Taking jj, as unit, the following magnifications are recom- 
mended : 



* Bull. Soc. Bot. France, 36 (1889) p. 207 



THE BACTERIOLOGICAL WORLD. 



213 



Magnifica- Coefficients, 

tions in ii 
2500(dimens. of the figure) divided by 2.5=n jx (absolute value). 



2000 
1500 
1000 
500 
250 
200 
125 
100 



2 =n jx 

" l,5=n m 

multiplied by 1 =n jx 

—n jix 

=n pi 

=n JX 

=11 fX 
■-71 IX 



10 =1 



Only those coefficients are practicable which, multiplying 
or dividing 1000, give whole numbers as product or quotient. 
Magnifications greater than 1000 are obtained by multiplying, 
those less than 1000, by dividing. 1000 by the coefficient. The 
above scale, which represents the magnifications which it is in 
this way possible to express in whole numbers, will answer all 
the requirements of microscopical figures. — Journal of the 
Royal Microscopical Society, December, 1890, p, 787. 

• Influence of coloring matters on spermatozoa.* — Dr. 
Emma Leclercq gives the following tabular statement as to the 
effects of various reagents on spermatozoa (Jour. R. M. Society') : 



COLORING MATTER. 



Carmine alone ( Frenzel ) 

Carmine alone ( Flemming 

Renault's hematoxylin 

Ranvier's picrocarmine 

Ehrlich's violet and eosin 

Ehrlich's violet and carmine 

Picrocarmine and methyl-green . 



RESULTS. 



On Nucleus. 



On Accessory 
Corpuscle. 



Light carmine 

Rose 

Violet 

Rose 

Violet 

Violet 

Violet 



Pale Rose 

Red 

Deep violet 

Orange 

Rose 

Rose 

Carmine yellow 



( The spermatozoa being green ) 



Preserving lower organisms in microscopical prep- 
arations. — Pure blood serum is recommended by Dr. W. 
Migula as a suitable medium for examining and preserving 

* Bull. Acad. Roy. Belg., 60. (1890) p. 138. 



214 THE BACTERIOLOGICAL WORLD. 

delicate animal and vegetable objects. He uses the commercial 
blood serum, and filters in an ice box, through bibulous paper, 
frequently changed. The filtrate is mixed with 10 per cent, 
pure glycerine, and incubated at 45° to 50° C. When all the 
water has been evaporated, the glycerized jelly is preserved in 
stoppered vessels. When required for use, a small quantity is 
dissolved in 10 to 15 times its volume of distilled water, and a 
large drop placed on the slide. Into this drop the living 
organism is pipetted, and then the slide is placed in an 
incubator at about 50°, in order to thicken down the fluid. 
When of the right consistence, the cover-glass, moistened with 
a mixture consisting of 40 parts glycerine, 20 parts absolute 
alcohol and 40 parts water, is imposed. The preparation is 
again heated for a couple of hours, and then ringed round. — 
Journal of Royal Microscopical Society, December, 1890, p. 804. 



MISCELLANEO US. 



DISINFECTION BY SULPHUR. 

There has been much discussion recently respecting the 
efficiency of sulphur as a disinfectant for various infectious 
diseases, the efficiency of this method of disinfection having 
been denied by some physicians whose opinions have been 
widely quoted in the newspapers. Confirmatory of the results 
obtained by the State Board of Health of Michigan, we are 
glad to be able to' quote the following from a work by Dujardin- 
Beaumetz, entitled Les Nouvelles Medications : — 

"Twenty grams of sulphur to a cubic metre (1.53 lbs. 
per 1,000 cubic feet of air space) destroy the different micro- 
organisms in a moist state, but it is necessary to increase this 
dose if one wishes to destroy some organisms in a dry state. In 
fact since the last communications to the Academy, M. Bardet 
and myself, aided by M. Ohambou, have continued these exper- 
iments upon micro-organisms in a dry state, and particularly 
upon vaccine virus. We have taken from the pustules of 
vaccinia scabs which we have reduced to fine powder, and 
placed in chambers where were variable quantities of flowers 



THE BACTERIOLOGICAL WORLD. 215 

of sulphur. When a dose did not exceed 20 grams per cubic 
metre, the vaccine powder did not lose its properties, and one 
could, by inoculating animals and infants, obtain a vaccine 
eruption. 

"With 30 grams per cubic metre (2.297 lbs. per 1,000 
cubic feet of air space), the results obtained were uncertain, 
sometimes the powder losing its properties ; but when the dose 
is increased to 40 grams per cubic metre (3.06 lbs. per 1,000 
cubic feet of air space), the inoculations are always inactive. 
So, then, for vaccine, and probably for variola, if one desires to 
destroy the contagious ' germs ' in a dry state, it is necessary to 
double the dose of 20 grams which we have already fixed. 

"'According to the experiments of Yallin and of Legouest, 
20 grams are sufficient for typhoid fever, while, according to 
Vallm, 40 grams are necessary for the microbe of tubercu- 
losis." — Hall's Journal of Health. 



TREATMENT OF DIPHTHERIA BY IODIDE OF POTASSIUM. 

S. Zinenko has obtained very encouraging results by the 
internal administration of iodide of potassium in diphtheria. It 
is administered to adults in doses as high as sixty grains a day ; 
children aged from one to fourteen years are given, each day, 
twice as many grains as they have years. The remedy is con- 
tinued until the appearance of symptoms of iodism, which is 
generally noticed from the second to the fourth day of the dis- 
ease. In case of cardiac weakness, tonic treatment is given at 
the same time. The author uses neither applications nor 
cauterization to the throat, as he considers them both irritant 
and useless. On the other hand, he employs the atomizer with 
a solution of boric or salicylic acid of from two to three per 
cent. The same is also to be used as a gargle. With children 
he uses lavage of the pharynx, according to the method of Dr. 
Stegenski. All the patients receive frictions of mercurial 
ointment in the region of the sub-maxillary glands (twenty-two 
grains, twice daily). Toward the end of the treatment, hydro- 
chlorate of quinine is given for its tonic effect. This method 
has been applied in all to twenty-eight persons. Some of the- 
patients died. At the same time, in another service of the 



216 THE BACTERIOLOGICAL WORLD. 

hospital, where the patients are treated by other methods, 
sixteen deaths occurred out of twenty-nine cases. In another 
hospital, where the hygienic surroundings are better than in the 
service of the author, during the same epidemic, the mortality 
was fifty per one hundred. — Bull. Gen. de Ther. 



SMALL-POX. 



Permit me to call attention to a plant which I have had fre- 
quent opportunity to prescribe in small-pox, and which possesses 
more remarkable virtues than any other remedies in the treat- 
ment of variola. I allude to saracenia purpurea which grows 
in North America and the common name of which is pitcher 
plant, huntsman's cup, etc. It has the power to abort the 
variola, and to cure it in a few days. If it is given during the 
period of incubation the disease will not be developed. If it is 
given when the eruption is fully out, a few doses will cause the 
papules to become pale, to diminish in size, and to disappear so 
completely that in ten days no trace of them will be left. — Dr. 
P. Balmoussieres in Clinique. 



MICROBES IN OTITIS MEDIA. 

Drs. A. Maggiora and G-. Gradenigo ( Gentralbl. fur Baht. 
u. Parasitenh., Band 8, No. 19, October 30, 1890 J mention 
the results of researches of previous authors, and then give 
their own methods of examination. They find that by cover- 
ing and plugging the end of a silver sterilized Eustachian 
catheter, it may be passed into the Eustachian tube, without 
becoming contaminated by the nasal mucus. The plug is 
withdrawn by means of a thread, and a celluloid bougie — which 
had been sterilized by repeated energetic rubbing with steril- 
ized cotton wool — is introduced for at least 1 centimetre into 
the tube. The bougie is withdrawn, and then placed in tubes 
containing gelatine and agar, and plate cultivations are pre- 
pared in the ordinary manner. In thirteen cases of chronic 
middle ear catarrhal inflammation with thickened membrane, 
fifteen ears were examined with the following results : Micro- 
coccus candicans was found in six out of the fifteen cultiva- 
tions ; saccharomyces roseus in four ; saccharomyces ellipsoideus, 



THE BACTEBIOLOGICAL WOKLD. 217 

bacillus subtilis, micrococcus cereus albus, penicilhum glaucum, 
aud diplococcus citreus conglomeratus in two each ; sarcina 
lutea, micrococcus citreus (II), bacillus albus, micrococcus 
cereus flavius, sarcina alba, merismopedia aurantiaca, and 
micrococcus opalescens in one each. In one case nothing was 
found, and in one non-pathogenic organisms that liquefied 
gelatine were met with. The authors conclude that in the 
later chronic or dry stages of middle ear inflammation there is 
no evidence of the presence of pathogenic organisms, as all 
those mentioned above may be looked upon as entirely innoc- 
uous. As regards the earlier stages, although there is nothing 
proved as yet, they consider it is quite possible that specific 
infective organisms may play some part, as their experiments 
are not at all inconsistent with such a condition. — British 
Medical Journal. 



TUBERCULAR INFECTION FROM MILK. 

Courtenay De Kalb, writing to the Nation from Eastern 
Peru, says : " Cattle are raised here solely for beef, milk never 
being used. It appears that in the tropics milk is even more 
likely to be infected with the bacillus tuberculosis than in the 
north, and so many cases of consumption have been traced to 
its use, that the entire population, with scarcely a single 
exception, leaves it absolutely alone. — Druggists Circular. 



218 THE BACTERIOLOGICAL WORLD. 

REMEDIES AND CLINICAL NOTES. 



ABORTIVE TREATMENT OF HERPES, AFTER PROF. LELOIR. 

J$_ Resorcine 3 ss - 

Alcohol 90° § iij & 3 j. 

If the pain is very severe he recommends : 

I£ Hydrochlorate of Cocaine grs. xv. 

Ex. Cannabis indica. 

Essence of Mint aa 5 iJ ss * 

Alcohol 90° • g iij & 3 i. 

Cover the diseased parts with impermeable cover, then dampen a 
light cloth or gauze with one of these solutions and apply it over the 
whole region. 

CURATIVE TREATMENT OF HERPES. 

}$_ Aristol 

Resorcine aa 3 ss * 

Tarro-petrolene (or Petroleum Com. 

No. 1) giij. 

Ft. Oint. 

Sig. : Apply twice daily over the diseased region. 

Fowler's solution of arsenic may be used besides, internally. 

URTICARIA. 

]£ Iodol 

Chloral aa 3 ss * 

Tarro-petrolene No . 1. g j. 

Ft. Oint. 

Sig. : Apply twice daily in gentle friction. 

Mr. Quinquaud advises camphorated alcohol with a little chloro- 
form and water in extreme itchiness. Internally he prescribes arseniate 
of soda. 

SORE MOUTHS OF NURSING WOMEN. 

I£ Katharmon f. % ii. 

Glycerini f . 3 "• 

Aquae Menth. Pip f. § ii. Misce. 

Sig. : Use as a mouth wash two or three times a day. 



THE BACTERIOLOGICAL WORLD. 219 

SYPHILITIC SORES IN THROAT, ETC. 

Jfc Listerine 5 1 to 3 

Glycerine % 2 

Salt grs. 5 to 10. Misce. 



Or: 



Ifc. Listerine ^2. 

Aquae § 10. 

Sodii Bicarbonate grs. 5. 

Sodii Biborate grs. 5. Misce. 

— Dr. Carpenter, in Medical Brief. 



QUERIES AND ANSWERS. 



Under this heading we will answer, as briefly as possible, short, 
concise, rational questions of subscribers, concerning bacteriological 
subjects only, as far as our knowledge may afford. 

Q. It is stated that pyoktanin is non-poisonous ; how then does it 
act as a germicide ? Sub. St. Louis, Mo. 

A. It seems to act by virtue of its staining properties ; it fixes 
itself on and throughout the body of bacteria, and thus destroys their 
vital energy. 

Q. What technical work would you recommend for use by a 
beginner in the study of bacteria ? 

A. There are several good books, and each has its special 
value. Get catalogues from medical book publishers. For one versed 
in histological technique, Salomonseri 1 s Bacteriological Technology is 
an admirable treatise. It is published by Wm. Wood & Co., New 
York. I would recommend beginners to read Prudden's " Story of 
Bacteria" published at 75 cents by G. P. Putnam's Sons, New York. 

Q. To what species of beings belongs the parasite of malaria, as 
given in the illustration of the February number ? 

D. V. S., Chicago, 111. 

A. From all that we know so far (the study is not complete), it 
seems that the parasite belongs to the gregarina, a genus of protozoa, 
family gregarinidae. 



220 THE BACTERIOLOGICAL WOELD. 

PUBLICATIONS. 



NOTICE OF SPECIAL PUBLICATIONS. 

BY PAUL GIBIER, M. D., 

Collaborator of The Bacteriological World. Director New York Pasteur 

Institute. 

IN FRENCH. 

Persistence of the Foramen of Botal ( interauricular orifice.) Soc. 
Anatom. Paris, 1880. 

On Tuberculosis of the Testicles. Soc. Anatom. Paris, 1880. 

On Gysts of the Ovary (j>atholog. anatomy). Soc. Anat. Paris, 1880. 

On Syphilis. Late Secondary Lesions. Soc.CUniq. Paris, 1881. 

On Syphilis. Unusual Situation of the Primary Lesions. Union 
Medicate. Paris, 1881. 

On Pulmonary Wounds by Fracture of the Clavicle. Soc. Clin. 
Paris, 1881. 

On Syphilis. Excision of the Initial Ulceration at its Beginning. 
Soc. Clin. 1881. 

On Typhoid Fever. Paris, 1881. 

Paralytic Symptoms Produced by Lightning. Soc. de Biologic 
Paris, 1881. 

On Typhoid Fever. Parasitic Spots. Soc. de Biolog. 1881. 

On Cerebral Tumor, etc. Soc. Anat. 1881. 

On Anatomical Pathology of the Right Heart. Soc. Anat. Paris, 
1881. 

Bacteria of the Pemphigus Acutus. Researches demonstrating 
that the Pemphigoid Fever is caused by a Specific Bacteria. Annates 
de Dermatologie. Paris, 1881. 

On Dermoid Cyst of the Fingers. Paris, 1881. 

On Enteroclysmus. Experiment. Research. Union Medicale. 1881* 

On Anthrax. Possibility of Infecting the Cold-blooded Animals 
by Artificial Elevation of their Temperature. Academie des Sciences. 
Paris, 1882. 

Action of Cold upon Trichina- In collaboration with Prof. H. 
Bouley, President of the Academie des Sciences. Paris, 1882. 

Hydrophobia (Experiment. Research on). Its Inoculation, 
Hereditary Transmissions, .etc. Academie des Sciences. Paris, 1883. 

Hydrophobia. Its Microbe. * Academie des Sciences. 1883. 

Hydrophobia. Effect of the Cold on the Conservation of its 
Virus. lb. 

Hydrophobia. Birds Contract and May Transmit it, but Re- 
cover, lb. 



THE BACTERIOLOGICAL WORLD. 221 

Hydrophobia. Experiment. Researches, etc. (One Volume.) 
Asselin & Houzeau, publishers, Paris, 1884. 

On Hydatid Pulmonary Cysts. In collaboration with Prof. V. 
Cornil of the Faculte de Paris. Soc. Anat. 1883. 

On Cholera. From a report presented to the French Government, 
which intrusted the author with the mission of studying the epidemic 
of France, 1884. Paris. ( Exhausted edition.) 

On Vaccination of Cholera. Experim. Research., in collaboration 
with Dr. Van Ermangem, of Gand. Acad, de Med., of Paris, 1885. 

Hydrophobia. Attenuation of its Virus" by Passage through the 
Organism of the hen. Soc. de Biolog. 1885. 

On Primitive Virulence of Venereal Adenitis. Soc. de Biolog. and 
Annales de Dermatologie. 1885. 

Report to the French Government on the German Laboratories of 
Bacteriology ( official mission). 1885. 

Report to the French Government on the Cholera in Spain (official 
mission). 1885. 

Psychical Researches. Spiritism and Fakirism. A Vol., 400 p., 
with engravings. Paris, 1886. 

On Disinfection by Steam without Pressure. Academie de Medi- 
cine. Paris, 1886. 

Yellow Fever. Experimental Investigations made in 1887-1888 in 
Havana. ( Official mission of the French Government.) Acad, des 
Sciences and Acad, de Med. Paris, 1888. 

Yellow Fever. Its Etiology ; Its Treatment. Bulletin Medical. 
Paris, 1888. 

Bacteriology. On a New Process of Cultivation in Matrasses in 
order to Isolate the Microbes. Acad, de Med. Paris, 1888. 

Consumption. On a New System of Inhalations. Acad, de Mede- 
cine, of Paris, 1888. 

Action of Cold upon Trichina. Refrigeration has no Effect. Acad, 
des Sciences. Paris, 1889. 

Analyse des Choses. ( Psychology.) One Volume. Paris, 1889. 

IN SPANISH. 

Fiebre Amarilla. Experiment. Research on Etiology and Treat- 
ment of Yellow Fever. Acad, of Sciences of Havana. Cronica Med. 
Quirurgica de la Habana, etc. 1888. 

Fiebre Amarilla. Sobre el tratamiento de la fiebre amarilla 
(Treatment of yellow fever.) Cronica Med. Quirurgica de la Habana. 
1888. 

IN. ENGLISH. 

Yellow Fever. Gaillard's Medical Journal, of New York, Feb- 
ruary and March, 1889. 

Anti-rabic Inoculations. Mechanism of Immunity. American 
Medical Association. 1890. 



222 THE BACTERIOLOGICAL WORLD. 

The Pasteur Treatment. North American Review, August, 1890. 

Dr. Koch's Discovery. North American Review, December, 1890. 

The Principles of Anti-rabic Vaccination. Bacteriological World, 
January and February, 1891. 

Three communications to the Berlin Medical Congress in 1890. 
First. A new theory about temperaments. Second. Peroxide of 
hydrogen anti-septic properties. Third. Hydrophobia. 



Senn's Principles of Surgery.— This is the clearest modern work 
of the kind that has come to our notice. Indeed it has a peculiar 
place to fill, which to this date was filled by the combination of a 
number of excellent works. It is a book of 600 pages, divided into 
twenty-four chapters, and is illustrated with 109 wood engravings. 
Several of these engravings are original. Among these may be 
mentioned the illustrations of nerve suture, ligation of veins, arteries, 
and the results thereof ; and above all the very striking picture of 
phagocytosis, bringing forcibly to the mind, the action of the white 
blood corpuscles on bacteria. All the other engravings are well chosen 
from a large number of authors. 

The most important feature of the work, from our point of view, 
is the information it imparts on the role of micro-organisms in relation 
to the principles of surgery. It is bound to do great good in this par- 
ticular line of thought and practice, and to influence useful modifi- 
cation of the treatment of surgical diseases. 

The price, cloth bound, is $4.50; sheep, $5.50 net. The publisher 
is F. A. Davis, of Philadelphia. 

Heredity, Health and Personal Beauty. — Under this title Dr. 
J.V. Shoemaker sends to the world a unique, fascinating book of great 
interest. It was intended, the author states, for the general public 
when he began to write it, but so much of a medico-scientific character 
has entered into it, that it contains a great deal that will benefit the 
physician and the scientific mind. 

It is divided into thirty-seven chapters, beginning with the general 
laws of health and ending with a chapter on " Household Remedies.' ' 
Certain chapters such as "The sentiment of the beautiful," "The 
source of beauty of the fair sex," " Grace, the crown of beauty," " The 
evolution of the American girl," are pleasing and instructive reading. 
They will repay a careful perusal better than the best and most 
instructive novel bearing on the subjects treated, 

The pith of the work consists of hygienic laws to be followed in 
order to preserve or perhaps become graceful, beautiful and healthy ! 

Ladies, do not miss it. Physicians, get it to teach your lady 
patients ! 

It is published by F. A. Davis at the price of $2.50 and $3.50 net 
in cloth or sheep binding. 



THE BACTERIOLOGICAL WORLD. 



223 



a.ktiseptic. 

Prophylactic. 

Deodorant. 



LISTER 




Non-Toxic 

Non-Irritant. 

non-escharotic 



FORMULA.- Listerine is the essential antiseptic constituent of Thyme, Eucalyptus, 
Baptisia, Gaultheria and Mentha Arvensis, in combination. Each fluid drachm 
also contains two grains of refined and purified Benzo-boracic Acid. 
DOSE.— Internally : One teaspoonf ul three or more times a day ( as indicated ) 
either full strength or diluted, as necessary for varied conditions. 
LTSTERINE is a well-proven antiseptic agent — an antizymotic— especially useful 
in the management of catarrhal conditions of the mucous membrane ; adapted to 
internal use, and to make and maintain surgical cleanliness— asepsis— in the treat- 
ment of all parts of the human body, whether by spray, irrigation, atomization or 
simple local application, add therefore characterized by its particular adaptability to 
the field of 

PREVENTIVE MEDICINE -INDIVIDUAL PROPHYLAXIS. 



* LISTEBIME * 

Destroys promptly all odors emanating from diseased gums and teeth, and will be 
found of great value when taken internally, in teaspoonful doses, to control the fer- 
mentative eructations of dyspepsia, and to disinfect the mouth, throat and stomach, 
It is a perfect tooth and mouth wash, indispensable for the dental toilet. 

Descriptive literature upon request. 

LAMBERT PHARMACAL CO., ST. LOUIS, MO. 



S MAW SON & THOMPSON, 

LONDON, E. C. 



AGENCIES: 

ROBERTS & CO., S. PAPPENHEIM, 

PARIS. BERLIN, W. 



VILANOVA HOS. Y CIA 

BARCELONA. 



A MAN IS KNOWN BY THE COMPANY HE KEEPS, AND 

A DOCTOR IS KNOWN BY THE LITERATURE HE READS." 

Moral : SUBSCRIBE FOR 

TheMedicalHerald 

The Progressive Exponent of Practical Medicine and Surgery. 

Read its Original Articles (photo engravings furnished free). 
Read its Reports of Hospital Clinics. 

Read its Society Reports from all parts of the West and South . 
Read its Special Correspondence from the Eastern cities. 
Read its seven departments of Medicine and Surgery by special writers. 



The handsomest and most original publication in the United States, and 

the best medium in the West for reaching an intelligent 

class of readers. 

SIX HUNDRED PAGES SOLID READING MATTER FOR $2.00. 

Send in your name at once and secure first numbers of the new series 

before they are exhausted. 

Address, DANIEL MORTON, M. D., 

MEDICAL HERALD, Editor. 

Saint Joseph, Missouri. 



224 THE BACTERIOLOGICAL WORLD. 

Self- purification of Flowing Water. — A reprint of a biological 
study on this subject, by Dr. Chas. G. Currier of New York, and 
which was published in the American Journal of Medical Sciences 
has reached our office. It is a very interesting and useful contribution 
to hygiene, containing, as it does, much information showing the 
influence of polluted water in the causation of disease. It contains 
a number of tables giving the ratio of bacteria per cc. in the water of 
many rivers and hydrants. The author deserves much credit for his 
splendid presentation of so many useful facts in so small a space. 

The Story of Bacteria, and Dust and its Dangers. — These 
two excellent little works of Prudden on bacteria, which we reviewed 
in February number, are published by G. P. Putnam's Sons, New 
York and London, at 75 cts. each. 



ITEMS AND NOTES. 



The St. Louis College of Physicians and Surgeons. — Dr. A. S. 
Barnes informs us that their " ad." in last issue is erroneous as to date. 
The Spring term begins March 15th, and the winter term the 10th of 
September. We are sorry to have been so misled by the manuscript 
as to publish the wrong dates. See advertisement on the last inside 
cover page. 

The St. Louis Clinique. — By mistake in our last issue, the notice 
of the improvement in the dress of this excellent medical publication 
was erroneous in its heading. It was entitled La Clinique instead of 
the St. Louis Clinique, as it should have been. 




DR. JHENEAGE GIBBES. 



m„ 



11JJ 



ACTERIOLOGICA 




A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies. 

Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological World, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to at the 
same office. 

Authors of original articles published in this magazine will be entitled to twenty 
copies of the number containing it, provided, that they request the same in writing 
when sending the communication. Reprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. 

Discontinuation :— At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first of next January. 

collaborators: 



Paul Gibier, M. D., New York. 
Prof. W. T. Belfield, M. D., Chicago. 
Prof. A. W. McAlester, A. M., M. D., 

Missouri University Med. Dep't, 
Prof. Henry P. Loomis, M. D., 

New York. 



Prof. L. Bremer. M. D., St. Louis, Mo. 
J. W. Stickler, M. D., Orange, N. J. 
Prof. Paul Schweitzer, Ph. D., 

Missouri University. 
Prof. T. J. Burrill, Ph. D., 

Illinois University. 



B.T. 


Galloway, B. S., Section of Vegetable Pathology, Washington 


D. C. 


Vol. 1, 


COLUMBIA, 


10, 


U, S. A, APRIL, 1891. 


No. 4, 





EDITORIAL. 



CONCERNING RACES AMONG BACTERIA. 

Mr. M. 0. Gessard in his work on biological chemistry at 
the Sorbonne and Pasteur's Institute, Paris, established that 
the pyocyanic bacillus, in its chromogenic function, produced 
a variety of pigments according to differences in the composi- 
tions of various culture media. His first investigations were 
made* with a micro-organism isolated from the dressing (of a 
wound). Afterwards he studied the functions of two other 
organisms of different origin. In the course of these studies 
Mr. Gessard investigated the measure, constancy and the ques- 
tion of necessity of relation between microbe and medium in 

* Annales de 1 Institute Pasteur, T. IV, page 88. 



226 THE BACTERIOLOGICAL WORLD. 

the production of pigment ; if these relations could extend to 
other germs of whatever source ; if exceptions could not spon- 
taneously occur ; or if forms could be provoked artificially and 
perpetuate themselves afterwards by way of generation. In 
other words he asked himself the question, can there be differ- 
ent races of the pyocyanic bacillus ? 

The idea of races among micro-organisms of the same 
species was introduced by Pasteur during his study of alcoholic 
fermentation (as far back as 1862 ). He demonstrated the dis- 
tinction between "high." and "low" yeast of breweries, prob- 
ably originating from a unique germ, "illustrating a new 
example of those modifications of plants and animals which 
have become hereditary by a prolonged domestication." 

Mr. Gessard has concluded from his investigations — and his 
conclusions are sustained by the careful experimentation of 
Wasserzug * — that races are established among germs. Both 
of these scientists have seen that these functions of pyocyanic 
bacilli and other germs may be modified, altered, changed in 
such a marked degree as to render their identification with the 
primitive parent organism difficult without a long series of cul- 
tures in various conditions and media. Indeed, we have seen 
ourselves that even the forms may be changed by surroundings 
and nourishment. 

A same species, in course of development and successive 
generations may, according to variation of external influences, 
nutrition and medium of existence, produce a number of germs 
different in physiological functions, and sometimes slightly in 
morphology. These altered organisms may be afterward culti- 
vated and preserved with the acquired modification, seemingly 
by hereditary transmission. These microbes with different 
attributes, coming from an original common parent, are true 
races, and the knowledge of the process of evolution in produc- 
ing the alterations — a process that the eye can follow clearly, 
down the scale of degeneration or upward to a higher physio- 
logical condition, perhaps — offers much food for reflection con- 
cerning the evolution of higher species in the animal and 
vegetable kingdoms. The truth of the profound changes 



* Annates de l'lnstitute Pasteur, T. I. & II. 



THE BACTERIOLOGICAL WORLD. 227 

produced gradually in a given organism by the surroundings, 
climate, medium of existence, food, etc., which eventually 
become physiological characters * susceptible of reproduction in 
descendants are apparent to all observers ; but here are little 
individual unicellular organisms as full of life as more complex 
beings, in which we can follow the evolution step by step with 
the eye. 

When we consider that a microbe is thus a unique cell in 
which life is centered, and that a larger organism — man say — is 
an aggregation of physiologically related cells, individually 
endowed with life as the single micro-organism is, we grasp the 
application of appreciable microbic evolutionary phenomena to 
more highly organized beings. Are not, in fact, unicellular 
and multicellular beings closely related physiologically ? 

From a single kind of bacteria, viz., the pyocyanic bacillus, 

taken in the dressing of a wound,- Mr. Gessard produced, in a 

series of cultures in different media, carried on successively 

more than one year, four different races with the following 

properties : 

f Ajgiving pyocyanine and fluorescence 

„.-„,,, .-©IB, giving only pyocyanine. 

Species, Bacillus Pyocyanic « { ~ . , . „ , 

J J «| C, giving fluorescence only. 

|^D, no fluorescence nor pyocyanine. 



VACCINATION AGAINST DIPHTHERIA. 

Mr. Fraenkel,f following his researches on the poison of 
diphtheria, made jointly with Mr. Brieger, has been seeking a 
mode of vaccination by means of attenuated cultures, and by the 
use of the toxic products of the microbe of diphtheria. 

The best results were obtained with a liquid culture heated 
an hour at 149 to 158 F. About ten cc. of such a culture, aged 
three weeks and treated as we have said, confers immunity 



* Mutilations, anatomical sequestration and the like are not con- 
sidered changes in the sense implied here. 

f Berl. Klin. Woch. No. 49, 1890. 



228 THE BACTERIOLOGICAL WORLD. 

against subsequent inoculation of the most virulent virus, on 
condition that the latter be not practiced sooner than fourteen 
days after vaccination. 

When we reflect a moment on the fearfully toxic properties of 
the product of this bacillus ; the fact that it lives chiefly on the 
mucous membrane, whence the poison generated is introduced 
into the circulation ; that this poisonous matter is' the greatest 
factor m causing pathological phenomena and death, we must 
admit that vaccination against diphtheria is a most difficult 
problem to solve. Still, some day, doubtless, diphtheria will 
be prevented some way. 



THE STERILIZATION OF MILK. 

The first endeavors to sterilize milk were made chiefly 
from a commercial standpoint, i. e., to preserve the fluid sweet 
a longer time. But, as the knowledge of pathogenic germs and 
the action of ferment organisms became clearer, the suggestion 
naturally entered the medical mind to sterilize milk from a 
hygienic and therapeutic point of view, chiefly for the nourish- 
ment of infants in health or disease. 

A large number of experiments have been made by various 
experimentalists, with a view to discover the most practicable 
means of sterilization combining effectiveness, cheapness and 
harmlessness. 

To that end, drug after drug, gas after gas, were tried in 
vain ; we have still to rely on the action of heat, though per- 
haps electricity under proper conditions may prove successful. 
But for every home, for every physician, heating is the only 
practical process. 

To kill all the bacteria usually found in milk — and there 
is always a wonderfully great and diversified army of them — it 
requires heat a few minutes about 224° F. A special steam 
sterilizer is necessary to attain such a high temperature. The 
same effects may be obtained usually by boiling the milk about 
thirty minutes. Heating a few minutes between 158° and 176° 
is sufficient to destroy temporarily the energy of a large number 
of milk microbes including several disease germs, but does not 



THE BACTERIOLOGICAL WORLD. 229 

hill many of them, and is not, therefore, sufficient as a pre- 
servative means. This is the temperature (about 167°), the 
application of which is known as Pasteurization. 

Unfortunately for the people, there are yet some medical 
men who will not concede any dangerous properties to microbes, 
and a vast number who are in ignorance of the action of germs 
in milk before or after ingestion. 

Is it only to destroy disease germs that sterilization is 
indicated ? By no means. Indeed, in infant-feeding, it is 
chiefly to prevent the irritative action of acid produced by 
microbes or germs of all kinds, whether truly milk ferments, 
pathogenic bacteria, or organisms harmless under ordinary 
circumstances. 

Milk coagulates, acidifies, under the influence of vegetative 
phenomena of various ferment organisms. The acid produced 
has the most baneful action on the delicate stomach and 
intestinal canal of the child. Sterilization of fresh milk 
destroys the property of the germs before any acid is produced 
by their process of nutrition. 

Besides the true milk ferments of various kinds, there is a 
number of cocci and bacilli which, grow in milk. Among the 
former, there are many which produce acids. Hence they, too, 
are deleterious and dangerous in the milk fed to the young. 

Duclaux, in his researches on the subject, noted that milk 
sterilized at about 167 Q F. was about free from acid-producing 
germs, but still contained live bacilli : a class of microbes 
apparently more difficult to alter, acting chiefly on the casein, 
and producing, as a rule, alkalinity instead of acidity. This 
fact suggested to the mind, that even though the heating of 
milk be not carried to boiling point it may be very useful in 
arresting the growth of the acid ferments, and leaving therein 
the bacilli, causing, by their nutrition, alkalinity of the media. 
We all know how frequently lime water is administered to 
bottle-raised babies to counteract the effect of acidity. Theo- 
retically we might, on the same principle, ascribe a beneficial 
influence to the alkaline-producing microbes left in the milk 
by pasteurization, — influence, which may be doubly useful 
because these organisms live at the expense of, and, therefore, 



230 THE BACTERIOLOGICAL WORLD. 

reduce, the casein which, in cow's milk, is always in excess of 
the necessary quantity for very young children. 

In the careful, constant use of pasteurized milk there is 
neither colic, green diarrhoea, or other intestinal disorders so 
common when using raw cow's milk ; but if the milk be highly 
heated, say at boiling point for several minutes, it is cooked 
and the casein is affected ; it digests well, but there may be 
some slight colic owing to the hardness of the foeces, and con- 
sequent interference with the naturally easy and frequent 
evacuation of the bowels at a tender age. But there are not any 
other intestinal troubles. 

Pasteurized milk, however, should be used early, for it will 
not keep long, the germs not having all been permanently 
arrested ; therefore, only careful methodical people can safely 
utilize it. Thorough sterilization on the other hand is practi- 
cable by all who may provide a vessel of some kind to boil 
water. To heat the milk at any temperature, it may be put in 
nursing bottles, plugged immediately with clean cotton wool 
(absorbent cotton), and then the bottles should be placed in a 
water bath, allowing the water and bottles to heat simultane- 
ously. There are more convenient steam sterilizers manu- 
factured now for the purpose. 

We have caused the use of both sterilized and pasteurized 
milk in' a number of bottle-raised children in the last two years 
with marked success, and we would recommend to doubting 
physicians the careful study of fermentation, coagulation and 
digestion of milk, and the effect of pasteurized, sterilized 
and raw cow's milk on the sensitive digestive organs of the 
young. 

It must not be forgotten, either, that the germs of cholera, 
typhoid fever, fibrinous pneumonia and others are destroyed or 
arrested in their pullulation by a temperature not higher even 
than that used in pasteurization, i. e., 158° to 167° F. 



THE TRANSFUSION OF BLOOD AND BLOOD SERUM IN 
MICROBIC DISEASES. 

The transfusion of blood between identical species, and 
even the transfusion of the blood of sheep to man, had been 



THE BACTERIOLOGICAL WORLD. 231 

tested and practiced without accident long before the bacteri- 
cide properties of blood serum were discovered. Then the 
operation was resorted to as a tonic treatment, — a method of 
restoring vital elements to debilitated, weakened organisms. 
Now it is tested chiefly to counteract the action of bacteria. 
Needless to say, that, as a germicide, it retains its nutritive 
properties. Unfortunately, it was found that the transfusion of 
an animal's blood directly into man's circulatory organs can be 
practiced only to a very limited extent, owing to pains and reac- 
tion resulting therefrom, whilst blood serum alone may be 
injected in a larger quantity without much danger. Conse- 
quently, the antiseptic benefits theoretically expected from the 
action of transfused blood in its natural condition can scarcely be 
realized. If blood serum acts as germicide and the other elements 
as nutritive, blood, pure, would seem preferable in debilitating 
diseases as tuberculosis, for instance, and it is unfortunate they 
cannot be as effectively used combined. In veterinary prac- 
tice, however, this objection may not hold good. 

At first, blood serum was injected into the system hypo- 
dermically, but this method limits its administration to a very 
small, insufficient quantity, owing to painful swellings and 
local disorders. Doses of O.oOcc. cause persisting pains when 
injected beneath the skin in man, whilst he may receive easily, 
without subsequent accident, lOOcc. in a vein. 

As a treatment for infectious diseases, it becomes evident 
that the serum used must proceed from an animal refractory to 
the disease to be treated ; it must be the serum of blood theoret- 
ically or positively known to be inimical to the causative 
microbe. 

With that thought in mind, Dr. St, Hilaire and Dr. 
Coupard injected serum of dog blood into the trachea in tuber- 
culosis. But dogs, though usually immune against this disease, 
are occasionally found suffering from it ; tubercles may develop 
in their system ; their blood, therefore, cannot be considered 
an absolutely reliable bactericide in the case under consideration. 
Hence, the trials with this serum are not to be looked upon as 
positively reliable. 

The next serum tried by various experimentalists, in the 
same disease, was taken from the blood of the goat, a species of 



232 THE BACTERIOLOGICAL WORLD. 

animals more refractory to tuberculosis than the dog. Prof. 
Lepine, of Lyons, France, following on the footsteps of Messrs. 
Picq and Bertin, of N antes, France, who had transfused goat's 
blood in man before, made numerous injections of serum into 
consumptives. 

The serum was quickly separated, aseptically, by means of a 
centrifugal process, filtered in aseptic filter, placed in a ster- 
ilized cylindrical vessel with a funnel-shaped aperture at 
the bottom, to which a sterilized rubber tube is attached. At 
the other end of the tube is a spring compressor closing it. 
A fine sterilized canula is fixed carefully at this end, the serum 
allowed to fill the tube to its very point in the manner of a 
hypodermic syringe, and it • is introduced carefully into the 
vein, in the direction of the flow, of course. The fluid must 
penetrate exceedingly slowly under the penalty of serious sudden 
reaction ; if allowed to trickle in very slowly, the quantity 
introduced may reach 88cc, to lOOcc. If some uneasiness, a 
reaction in other words, manifests itself during the operation, 
discontinue it. Injections may be repeated every two or three 
days, a thing impossible in hypodermic injections, owing to local 
pathogenous results. 

The speed of intravenous injection may be regulated 
readily, by lifting or lowering the supplying vessel more or less 
above the arm in which the canula is introduced.* 

The numerous experiments of Dr. Lepine, on this method 
of serum infusion, are very encouraging as far as practicability 
and safety is concerned ; it has many undoubted therapeutical 
advantages over the more simple hypodermic practice, but it 
remains to be seen whether the serum of goat blood is curative 
in tuberculosis. 

However, Dr. Bompard, Physician in Chief of the General 
Hospital of Vitry-le-Francois, relates the following results of a 
recent trial : 

" The patient had tuberculosis in the last stages — the 
third degree ; was so feeble that he could not rise ; appetite so 
altered that he ate scarcely anything and retained nothing of 
the ingesta with the exception of a few teaspoonfuls of coffee. 

^Description of apparatus in detail may be found in pages 146 and 
147 of La Tribune Medicate, No. 10, March 5, 1891. 



THE BACTERIOLOGICAL WORLD. 233 

Kespiration was labored, the surfaces cyanosed, and there was a 
severe pain of the scapula-humeral articulation preventing all 
motion of the arm. Forty grams of goat serum were trans- 
fused into the right cephalic vein. 

" Immediately the face assumed a more pronounced 
cyanose tinge, nearly black ; respiration worse than before ; 
pulse feeble ; a cold perspiration covered the face. These 
alarming symptoms lasted only a very few seconds, and all 
returned to previous condition. 

"■ In less than two hours afterwards he felt a relief such as 
he had not experienced for a long time ; the pain of the 
shoulder had completely disappeared ; the pulse was fuller, 
more regular, 100 per minute. Rectal temperature never rose 
above 38 any time during the day. The first urine passed was 
highly colored, almost dark. 

11 The condition since is truly remarkable ; appetite, excel- 
lent ; digestion, easy ; strength, increasing, the patient having 
risen from his prostrated condition and being able to walk 
about freely. Cough and expectoration have considerably 
diminished ; cyanosis and depression have completely disap- 
peared. All points, in fact, to a radical cure. Still, it is a 
first case ; will it prove a deceptive exception ? " 



GREEN BACTERIACEES. 

Gradually, though with extreme slowness, the gap between 
bacteria and the next forms in higher organization is closing. 
The naturalists, with laudable energy and perseverance, are 
filling the space, narrowing the distance, that separated the 
species. Already we can appreciate the connecting links 
between the microscopic species, and in time we will see, 
probably clearly, the chain of anatomical and physiological 
relation which binds all living organisms in their respective 
kingdoms. 

Mr. Van Tieghem described, early this year,* two bacteria 
presenting a green color, supposed to be chlorophyl : The 
bacterium viride and the bacillus vireus. 



* Note to the Academy of Sciences, January 26, 1891 



234 THE BACTERIOLOGICAL WORLD. 

More recently* Mr. P. A. Dangeard, Chief of the Botanical 
Department of the Faculty of Caen, while studying the algse of 
soft water, noticed two forms which, " by their behavior and 
by their endogenous sporulation," fixed his attention. They 
were constituted by very fine green filaments entangled with 
one another, apparently devoid of branches, ramifications and 
partition walls. The content was hyalin, non-granular, and 
slightly tinged with green, particularly at the moment of 
sporulation. The chlorophyl, if such is the green matter, was 
evenly dissolved in the protoplasm. 

In process of sporulation some filaments had as many as 
ten elliptical spores, some of them in groups of three or four. 
These spores varied between Qjj. and Spi in length, and presented 
a breadth of about 3jj. 

The spores, though formed after the manner of the bacter- 
iacees, do not present absolutely the same features as are known 
to exist in the common forms of this class of beings. For 
instance, they do not begin as dark spots, developing at the 
expense of reserve material of the mother cell, but "it is 
the entire mass of the protoplasm of the swelled spot which 
contracts, becomes more and more refringent, and surrounds 
itself by an envelope. 

Klein had observed this kind of sporulation in five forms, 
which he classed among the genus bacillus, f 

Mr. Dangeard proposes, for the forms he describes, the 
genus euhacillus, to impress their close affinity with bacilli. 
He believes that Klein's forms are of the same order. 



HENEAGE GIBBES, M. D. 

Dr. Heneage Gibbes, Professor of Pathology, University of 
Michigan, is the grandson of Sir George Gibbes, physician 
to Queen Charlotte. Dr. Gibbes commenced life as a sailor, 
and commanded a gunboat in the Chinese navy for some years. 



* Journal de Micrographie, February, 1891. 
f J. L. Klein, Botaniske Bacterienstudien. 



THE BACTEKIOLOGICAL WORLD. 235 

On giving up the sea, he entered the medical profession, grad- 
uating at the University of Aberdeen, and afterwards at the 
Royal College of Physicians, London. He commenced the 
study of normal histology under Dr. Klein before he grad- 
uated, and continued this study until he was appointed curator 
of the Anatomical Museum, King^s College, London, where he 
remained three years and made a thorough study of Morbid 
Anatomy. He was then invited to take the Chair of Physi- 
ology, Normal and Morbid Histology at the Westminster 
Hospital, London. In 1884, Dr. Gibbes was sent to India by 
the British Government to make an investigation into the 
etiology of Asiatic cholera. 

In 1887, the University of Michigan determined to institute 
a Chair of Pathology, which was offered to Dr. Gibbes, who 
accepted the position, and arrived in Ann Arbor in 1888. Dr. 
Gibbes was also for nine years physician to the Metropolitan 
Dispensary in the City of London. During his medical career 
he has published a large number of original works. He 
commenced an investigation into consumption in 1882, and in 
1883 he was requested by the local Government Board of 
London to carry on this work for them, the results being pub- 
lished in their reports. On arriving in this country, he lost no 
time in continuing this work, and enlisted Dr. Shurly, of the 
Harper Hospital, Detroit, to act with him. Their efforts have 
resulted in the discovery of a successful treatment for the disease. 

Dr. Gibbes has not confined his investigations to consump- 
tion, but has published many papers on other diseases. He 
has been engaged for some years in the investigation into the 
pathology of cancer, and hopes soon to find a means of arrest- 
ing that terrible disease. 

Bacteriological technique the world over is greatly indebted 
to this savant, and medical science has in him a most faithful, 
honored and exceedingly useful apostle. Prof. Gibbes stands 
high in the front rank of pathologists and bacteriologists of the 
world. 



236 THE BACTERIOLOGICAL WORLD. 

LESSONS IN BACTERIOLOGY. 



LESSON FOURTH.— PART I. 



BY PAUL PAQUIN, M. D. , V. M. 



HISTOLOGICAL TECHNIQUE. 

Before the study of bacteria can be profitably undertaken 
a fair knowledge of histology and pathology is necessary, and 
the essentials of microscopical technique as applied to botany 
should have been acquired. 

The students should be familiar with the methods of preser- 
vation of tissues, section cutting, staining, mounting and the 
manipulation of the microscope. In out' lesson on bacteriolog- 
ical technique we will suppose these subjects known. 

The microscope should have a solid base and be steady; a 
light instrument vibrates too much at the least shaking of the 
building, floor or table supporting it, particularly when the 
latter are themselves light or unsteady. 

The microscope should be provided with immersion lenses, 
(oil and water), and an Abbe condenser with a wide angle of 
aperture. There are a number of reliable houses furnishing 
excellent stands and lenses in this and other countries. Abbe 
and Zeiss, of Germany, manufacture, with a special kind of 
glass, apochromatic lenses which allow stronger oculars, give 
better results m photography, and present clearer pictures. 
Still, the best lenses of other houses are often equal to Zeiss 
apochromatic lenses, and much less expensive. 

As in other microscopical studies, a low power should be 
used in the first survey of the field. A magnification of 50 to 
200 diameters with an Abbe condenser, and a diaphragm with 
narrow diameter will answer the purpose. In thus examining 
the topography of the sections or other preparations, the 
stained bacteria may be located by their fixed color, particularly 
when they are numerous and grouped. 

In preparations doubly stained, i. e., with the field one 
color and the microbes another, the repartition of the latter is 
comparatively easy to detect ; even the bacilli of tuberculosis 
may be located by such low powers, and large microbes, as the 
bacillus anthracis, may be individually appreciated. But, to 



THE BACTERIOLOGICAL WORLD. 237 

determine the form, species, physical peculiarities of bacteria, 
clear lenses and a magnification of 300 to 1,500 diameters or 
more are necessary. 

The study of unstained bacteria may be accomplished with 
high- power lenses and a diaphragm. 

The best oil for immersion is pure cedar oil, but impure, 
thin cedar oil is worthless. In lieu of this, a mixture of chloral 
hydrate and glycerine may be used to advantage. The lenses 
should be carefully wiped dry immediately after using, and the 
microscope covered with a glass bell to protect it from dust and 
other influences. 

Accessories. — Certain articles are indispensable as acces- 
sories in micrographic investigations of bacteria. 

First. To clean instruments and glass: — 

A chamois skin, soft and smooth. 

Old soft rags, — say old linen handkerchiefs. 

Second. To handle' specimens, etc.: — 

Steel needles, ( ordinary sewing needles mounted in wooden 
handles ). 

Platinum wire needles mounted in glass rods. 

Fine forceps with spring, steel or brass. 

Spatulas, very thin, mounted in wood. 

Pincers with fine, toothless points. 

Scissors, fine, curved and straight, for dissection. 

Glass rods with one rounded extremity and a more pointed 
one. 

Third. To keep reagents, stains, etc.: — 

Flasks, small, with ground mouth for alcoholic solutions 
and acids. * 

Flasks, small, with pipettes for stains and reagents used in 
manipulations. 

Bottle cedar oil. 

Watch glasses and similar dishes, such as small crystal or 
porcelain salt-cellars, for staining, etc. 

Fourth. Essentials to mount: — 

Glass slides, ground edges, — an assortment of different 
thicknesses. 

Cover glasses, round and square, various thicknesses. 

Turn table to make rings on slides for special preparations. 



238 THE BACTERIOLOGICAL WORLD. 

Small labels for preparations. 
Slide boxes to preserve mounts. 

Fifth. Useful in mounting : — 

Black glass plate with smooth surface on which to mount. 

White porcelain plate for same purpose. 

Rubber bulb pipettes. « 

Camel's hair brushes. 

Hand-bulb blow-pipe to dry objects. 

Fine white tissue paper (cigarette paper ). 

Clean white bibulous paper. 

Microtome. — In histology and botany, satisfactory sections 
may be made by a careful, steady and experienced hand with a 
smooth razor ; but to diagnose bacteria in sections of tissue 
necessitates such extreme thinness that few can succeed by free- 
hand cutting. To obviate the difficulty, various accurate 
mechanical microtomes have been invented, and are kept in 
stock by optical houses ; satisfactory results are obtained by 
their use. 

To study bacteria in fresh tissues it becomes necessary to 
freeze the specimens in order to make sections. This is done 
with ether by the use of a mechanical device accompanying 
certain microtomes or sold separately. 

Camera-lucid a. — This instrument is needed for accurate 
drawings of microscopic fields. Drawing .pens, pencils, India 
ink, water colors, brushes, etc., etcT, should, of course, be also 
at hand. 

PART II. 

CHEMICALS, FORMULAS, ETC. 

The bacteriologist or student should always have in stock a 
certain amount of reagents, stains, etc., necessary to preserve, 
harden, decalcify, incorporate, fix, stain, mount and examine 
specimens. 

Stock of chemicals, etc.: — 

(a) Distilled water, renewed frequently, kept in clean 
stock bottle with rubber tube for exit as represented in Fig. 
4, 




Fig. 4. ( from crookshank. ) 

Glass jar or flask for distilled water for mounting purposes. This 
reservoir is provided with a glass tube air-inlet ( which, if plugged with 
cotton wool, will admit the air free from germs ); a rubber tube syphon 
with glass dropper and pincers allowing the exit of water in a fine 
stream. 




Fig. 5. ( from cornil and babes. ) 
Flask for Sterilized water. 



THE BACTERIOLOGICAL WORLD. 239 

(a) Sterilized water. See Fig. 5. 

( b ) Acids, such as nitric, muriatic, sulphuric, acetic 
3tc. 

( c) Alkalies, as caustic potash, carbonate of soda, ammo- 
lia, acetate of potash, etc. 

(d) Aniline oil used to make aniline water, toluidine 
ised in the same line. 

(e) Alcohol, absolute and diluted. 

(/) Cloves oil, or essence, spirits of turpentine ; essence 
)f bergamot, oil of origanum. 
0?) Xylol. 
( h ) Canada balsam. 
( ? ) Asphalt gum. 
(/) Hollis glue. 

Standard formulas for staining mounting, etc. — The 
!ollowing formulas have been adopted by bacteriologists, and are 
ised in various circumstances which will appear as we proceed 
.n the explanation of different methods of investigation, etc. 
Iliey should be prepared shortly in advance, in small quantities, 
so as to have no old depreciated stock on hand. The fresher 
:he preparations the better as a rule. 

Celloicline Solution : — Celloidine, ether, alcohol, equal 
Darts. 
Ebner's Solution: — (parts.) 

Muriatic acid,, , 5 

Alcohol 10 

Distilled water 20 

Chloride of sodium 5 

Glycerine Gelatine (Klebs): — 

Gelatine ( 1st. quality and well 

washed) ... 10 

Glycerine 10 

Phenol, a few drops. 
Add a little distilled water to the gelatine, let it swell, 
3mpty the excess of water, melt at mild heat and add the 
glycerine and then the phenol. 
Kleinenberg's Solution : — 

Aqueous ? saturated picric acid sol 100 

Sulphuric acid strong 2 to 5 



240 THE BACTERIOLOGICAL WORLD. 

Filter and add 

Distilled water 300 

Mutter's Fluid: — 

Bichromate of potash 2 

Sulphate of soda 1 

Distilled water 100 

Osmic Acid : — 

Distilled water 100 

Osmic acid 0.5 

This must be kept from light, in a bottle painted black, 

say, or surrounded by dark paper pasted. 

Alcohol 60 per cent. 

Acidulated Alcohol : — 

Alcohol : 100 

Muriatic acid 100 

Calamine and Alum ( Grenadier ): — 

Carmine 1 

Alum Solution, a 5-per-cent 100 

Boil twenty minutes and filter when cool. 

A niline Water : — 

Distilled water 100 

Aniline 5 

Shake and filter the emulsion. 

Bismarh Brown : — 

(a) Concentrated solution made in equal parts of water and 
glycerine, 

(6) Aqueous Solution: 

Bismark Brown 2 

Alcohol 15 

Distilled water 85 

Borax Carmine : — 

Borax 4 

Distilled water 100 

Add 4 per cent, of carmine, heat twice to boiling point, 
add equal parts of alcohol at 70 per cent., let settle for five hours 
and then filter. 



THE BACTERIOLOGICAL WORLD. 241 

Eosine: — 

{a) Alcoholic solution, saturated. 

( b ) Aqueous solution : 

Distilled water 100 

Eosine 5 

Fuchsine : — 

Fuchsine 2 

Alcohol 15 

Water 85 

Gentian Violet: — 

(_«) Alcoholic solution, saturated. 

(i-) Aqueous solution : 

Gentian violet — 2.25 

Distilled water : 100 

Solution of Gibbes for Double Coloration : — 

Take: 

Of hydrochlorate of rosaniline 2 

Of methyl blue 1 

Pulverize or crush in glass mortar; 

Dissolve aniline oil 3 

In alcohol rectified 15 

Add slowly to the first mixture and, last, 

Add distilled water slowly 15 

Preserve in a flask well corked. 

Hematoxylon Solution : — 

Hematoxylon 2 

Alcohol 100 

Distilled water 100 

Glycerine 100 

Alum. 2 

Iodine Solution: — 

Pure iodine 1 

Iodide of potassium 2 

Distilled water 50 

Iodine Solution ( Gram) : — 

Iodine 1 

Iodide of potassium 2 

Distilled water 300 



242 THE BACTERIOLOGICAL WORLD. 

Lithium- Carmine Solution (Ortli) : — 

Solution, saturated, of carbonate of lithium 100 
Carmine 2.5 

Magenta Solution ( GMes ) : — 

Magenta 2 

Aniline oil -. 3 

Alcohol (sp. gr. 830) 20 

Distilled water 20 

Methyl Blue :— 

(a") Alcoholic solution, concentrated, 

( l) ) Aqueous solution : 

Methyl blue 2 

Alcohol.. 15 

Water...... 85 

( c) Koch's solution : 

Concentrated, alcoholic solution of : 

Methyl Blue 1 

Potassium solution, 10-per-cent 2 

Distilled water 200 

( d ) Loeffler's solution : 

Concentrated, alcoholic solution of methyl 

blue 30 

Potassium solution, 1-10,000 .. 100 

Methyl Violet: — 

( a ) Alcoholic solution, coucentrated. 

( b ) Aqueous solution. % 

Methyl violet 2.25 

Distilled water 100 

( c ) Koch's solution : 

Aniline water 100 

Alcoholic solution of methyl violet 11 

Absolute alcohol 10 

Neelsen's Solution : — 

Dissolve fuchsine ... 1 

In aqueous solution of carbolic acid, 5-per- 
cent 100 

Add alcohol 10 



THE BACTERIOLOGICAL WORLD. 243 

Orchil ( Wedl ) :— 

Dissolve pure orchil without ammonia in : 

Absolute alcohol 20 

Acetic acid 5 

Distilled water 40 

until a deep red color is obtained, and filter. 

Picro- Carmine ( Ranvier ): — 
Dissolve : 

Carmine 1 

In distilled water 10 

And ammonia solution 3 

Then add : Saturated solution with filtered and 

cold picric acid 200 

Picro- Lithium Carmine (Orth): — 

To the solution of lithium-carmine mentioned 
above, add : 
Picric acid solution. 2.3 

Potassium Solution : — 

(a) 1 to 3 per cent. » 

( b ) 10 per cent. 
( c ) 33 per cent. 

Safranine : — 

(«) Alcoholic solution, concentrated. 
( b j Aqueous solution , 1-per-cent. 

Cliloride of Sodium Solution : — 

0.8 per cent. 

Vesuvine : — 

(a) Alcoholic solution, concentrated. 
( b ) Aqueous solution. 

Kuhne's Carmine : — 

Alcohol, 80 percent 50 parts. 

Muriatic acid 3 drops. 

Carmine q.s. 

Boil ten minutes and filter. 



244 THE BACTERIOLOGICAL WORLD. 

Differential Liquid : — 

Distilled Water , 10 cc. 

Sulphuric acid concentrated 2 drops. 

Oxalic acid, 5 per cent 1 drop. 

ZieliVs Stains for Tuberculosis : — 

{a) Alcohol, absolute 10 cc. 

Fuchsine ... 1 part. 

Carbolic acid 5 

Water. 100 

(6) Water 100 

Carbolic acid 3 

Alcohol 5 

Alcoholic solution of rubine. q.s. 

FraenheVs Solutions : — 

O) Distilled water 100 

Aniline oil : . 3 

Alcohol, pure 5 

( b) Aniline oil , . 30 

Nitric acid 20 

Alcohol, pure 50 

Methyl blue, to saturation 

Gibbes' Magenta Stain: — 

Magenta 2 grains. 

Aniline oil 3 cc. 

Alcohol 20 cc. 

Water, distilled . 20 cc. 

Dissolve the oil in the alcohol and shake. Eub crystals 
very fine with a little alcohol until all dissolved, then add dis- 
tilled water, stirring. Keep stoppered, labeled and dated. 

Acetate of Potash : — 

Concentrated solution. 

Farranfs Solution {Glycerine Gum): — 

Glycerine ^ 

Water y Equal parts 

Arsenic acid, sat. sol.... J 

Mix and add : Gum Arabic... , iy 2 parts 



THE BACTEKIOLOGICAL WORLD. 245 

Raulin's Fluid, Culture Media : — (parts.) 

Distilled water 1,500 

Cane sugar 70.0 

Tartaric acid 4. 

Ammonium nitrate 4.0 

Ammonium phosphate 0.6 

Potassium carbonate 0.6 

Magnesium carbonate . 0.4 

Ammonium sulphate 0.25 

Zinc sulphate 0.07 

Ferrous sulphate 0.07 

Potassium sulphate 0.07 

Pasteur's Fluid : — 

Distilled water 1,000.0 

Pure cane sugar 100.0 

Ammonium tartrate , 10.0 

Ash of yeast., ,.... 0.75 

Cohn's Fluid, ( as recently modified ) : — 

Distilled water 200 

Tartrate of ammonium 20 

Phosphate of potassium 20 

Sulphate of magnesium 10 

Tribasic phosphate of lime 0.1 

( Fifth Lesson in May number.) 



246 THE BACTERIOLOGICAL WORLD. 

ORIGINAL ARTICLES. 



KOCH'S TREATMENT OF TUBERCULOSIS. 
( From observations abroad.) 

BY PROF. WOODSON MOSS, M. D., 

Medical Department Missouri State University. 

It is with great reluctance that I attempt, at the earnest 
solicitation of the editor of this new and valuable journal, to 
give its readers an article upon the above all-absorbing subject. 

Having just returned from a three months - ' sojourn in 
Berlin and London, where I had gone, as the representative of 
the University of the State of Missouri, to investigate the use 
of Koch's lymph in the treatment of tuberculosis, I have 
thought, possibly, the best way of presenting an article would 
be to state what I saw and heard of this treatment, and how I 
was impressed with it. Upon arriving in Berlin, I found the 
hospitals easy of access, and the physicians, in charge, kind, 
courteous and untiring in their efforts to show visitors all they 
had in their hospitals. The opportunities to observe the use 
of Koch's lymph, or tuberculine, were greater than the ability 
of any visitor to take them all in. The hours were so arranged 
that you could spend nearly the entire day at the various clin- 
ical demonstrations. I was fortunate enough to get to Berlin 
after the great rush was over, and did not experience the dis- 
advantages of crowded clinics. If I should have been among 
the first to visit Berlin, I expect I should have gone there like 
many others, full of hepe of the impossibilities, and, like them, 
returned home discouraged and disappointed. But, as I did 
not go until the latter part of December, I had had time for 
sober, second thoughts, and, on my journey, I was constantly 
meeting or hearing the returned, giving their varied experi- 
ences. Some few were hopeful, but the majority were a little 
disappointed or skeptical, so that, when I arrived in Berlin, I 
might say that I was neither prejudiced for or against the 
treatment. 

One of the first things I did was to find out just what 
Koch claimed for his lymph, and not what the public were 



THE BACTEEIOLOGICAL WOELD. 247 

claiming for it. I think some of our journals and members of 
our profession have failed to make this all-important distinc- 
tion. I know by this date that most, if not all, of the readers 
of this journal have read KochV statements, but it will be nec- 
essary to restate them here if we expect to systematically dis- 
cuss and present this subject for their consideration: 

In what way this process occurs cannot, as yet, be said with cer- 
tainty, as the necessary histological investigations are not complete. 
But so much is certain that there is no question of a destruction of the 
tubercle bacilli in the tissues, but only that the tissue inclosing the 
tubercle bacilli is affected by the remedy. Beyond this there is, as is 
shown by the visible swelling and redness, considerable disturbance of 
the circulation, and, evidently, in connection therewith, deeply-seated 
changes in its nutrition which cause the tissue to die off more or less 
quickly and deeply, according to the extent of the action of the 
remedy. 

To recapitulate, the remedy does not kill the tubercle bacilli, 
but the tuberculous tissue ; and this gives us clearly and definitely the 
limit that bounds the action of the remedy. It can only influence 
living tuberculous tissue ; it has no effect on dead tissue, as, for 
instance, necrotic cheesy masses, necrotic bones, etc. ; nor has. it any 
effect on tissue made necrotic by the remedy itself. In such masses of 
dead tissue living tubercle bacilli may possibly still be present, and are 
either thrown off with the necrosed tissue or may possibly enter the 
neighbouring still-living tissue under certain circumstances. If the 
therapeutic activity of the remedy is to be rendered as fruitful as 
possible, this peculiarity in its mode of action must be carefully 
observed. In the first instance, the living tuberculous tissue must be 
caused to undergo necrosis, and then everything must be done to 
remove the dead tissue as soon as possible, as, for instance, by surgical 
interference. Where this is not possible, and the organism can only 
help itself in throwing off the tissue slowly, the endangered living 
tissue must be protected from fresh incursions of the parasites by con- 
tinuous application of the remedy. 

No one who has observed the use of the lymph in a large 
number of cases can deny that its power to bring about this 
necrosis is certain and about invariable. The few possible failures 
merely prove the rule. But as to the last of the above state- 
ment, viz.: "The endangered living tissue must be protected 
from fresh incursions of the parasites by continuous application 
of the remedy." This seems to be the first rub with some of 



248 THE BACTERIOLOGICAL WORLD. 

the opponents of the lymph. These opponents claim that the 
uninvaded tissues of the body are not protected by the contin- 
uous use of the lymph, and that this necrosis of living tubercu- 
lous tissue exposes the individual to the dangers of septic 
poisoning, accompanied by fresh invasion of the absorbed 
bacilli. To take a decided or emphatic stand for or against 
this view of the subject, when the use of the lymph is but a few 
months old, is, to say the least, a little premature. But, if we 
have to decide now, I claim that the weight of testimony is 
against the opponents of the lymph. Let us go to the crucial 
test of results, and see upon which side is reason. 

As the honored and distinguished Virchow seems to be the 
head and front of the opposition, and, as his position upon the 
subject has had more effect upon the opinions upon the use of 
the lymph than any other man, let us examine some of his 
statements, and see if they are as fair and just as we had a right 
to expect from one so honored and reverenced by medical men 
of all nationalities. I quote from the London Lancet of 
March 17, 1891: 

Limiting his remarks to facts determined by anatomical investi- 
gation, Professor Virchow said that, from the time at which the 
practice commenced until the end of December,, he had examined 
post mortem twenty-one cases in which, during life, injections of 
Koch's fluid had been practiced, and to these might be added some six 
or seven more examined during the present year. ( A large number 
also have been examined by his assistants in other hospitals, and else- 
where, and had yielded important facts.) 

There is an obvious difference between those cases in which the 
changes taking place in superficial parts can be observed clinically, 
and those where the internal organs are affected to an extent difficult 
to estimate by the most careful clinical investigation ; and it is of 
interest to contrast the latter with the former. Of the twenty-one 
cases referred to, sixteen were cases of phthisis in the limited sense, 
i. e., in which the lungs were especially involved. The remaining five 
cases comprised : First, a well-marked case of tuberculosis of bones 
and joints ; second, a case presenting the rare concurrence of cancer of 
the pancreas with small, smooth-walled cavities in lung and apical 
induration ; third, an empyema in a puerperal case, which would 
probably have proved fatal apart from the injections ; fourth, per- 
nicious ansernia, with very limited old lesions in lungs, and tuberculous 
pleurisy, and, fifth, a case of arachnitis tuberculosa. In the remaining 
sixteen, strictly pulmonary cases, all showed more or less extensive 
phthisical ulceration. 



THE BACTERIOLOGICAL WORLD. 249 

Reserving for another opportunity a detailed statement of these 
cases, Professor Virchow proceeded to make some general remarks on 
the experience derived from them. Just as observation of external 
parts had shown that the remedy acts as a severe irritant, exciting- 
redness and swelling, so the same effects had been well seen in the 
internal parts . He showed a preparation from a case of arachnitis 
tuberculosa in Henoch's clinic, in which there were also some old 
caseo-pneumonic areas in the lungs ( the probable focus of the menin- 
geal metastasis ), together with recent inflammatory changes. After 
four injections, amounting in all to two milligrams, the last being six- 
teen hours before death, the patient ( a boy two years and a half old ) 
died. Professor Virchow did not remember ever having seen so 
" colossal " a hyperaemia of the pia mater, and even of the brain itself. 
The pia matral vessels were extremely distended, and the brain tissue 
of a dull reddish tint. In this, the only case of the kind in the series, 
he personally examined the tubercles in the arachnoid, but could 
detect no degenerative changes in them ; they were well formed and of 
the ordinary character. Similar conditions of acute hyperaeruia and 
swelling were also to be found in other internal organs. They had often 
noted an unusual deep injection of the granulation, layers of old pul- 
monary cavities ; sometimes a heemorrhagic infiltration of the walls, 
or evidence of recent haemorrhage into the cavity. In a man, thirty 
years of age, with an old rectal fistula and numerous tubercular 
ulcers in the colon, death followed upon haemoptysis from an old ulcer- 
ating focus ; he had been injected seven times, the last occasion when 
the haemorrhage set in, being thirteen days before his death. But 
these visible changes were not confined to such marked hyperaemic 
swellings which obviously might be very transient, for there was not 
the least doubt that actual inflammation, with active proliferation, 
occurred to a considerable extent. This is well shown at the margins 
of ulcers and in the neighbouring lymphatic glands, especially the 
bronchial and mesenteric. The glands were most markedly swollen, 
that form of medullary swelling peculiar to acute irritation, and due to 
a rapid proliferation of the cells within the gland. This change is often 
associated with obvious leucocytosis, which may be assisted by the infil- 
tration of white blood corpuscles around the tubercle. Sometimes 
these inflammatory swellings are dangerous. Thus in the larynx, 
although the ulcers may become healthier, the neighbouring tissue 
enormously swells, and may produce a critical degree of stenosis. 
Sometimes the change may assume a phlegmonous type, comparable 
to what is seen in oedema of the glottis and retro-pharyngeal abscess, 
as illustrated in a case seen this month (January). No doubt it is 
difficult to assert that any such inflammation is due to the injections, 
for there is no criterion for any differentiation ; nor was he prepared 
to say precisely how this kind of inflammation may be recognized and 



250 THE BACTERIOLOGICAL AVORLD, 

distinguished from other inflammatory lesions arising in the course of 
phthisis. But in the lungs certainly there are lesions which are excep- 
tionally peculiar. 

Among the fatal cases of ulcerative phthisis the great majority 
showed widespread changes of recent date, mostly in the lungs, but 
also associated with pleurisy of a very severe type, simple and tuber- 
cular, liEemorrhagic and not rarely bilateral. The lung changes may 
be separated into two seemingly very different categories. The one 
corresponds to what is styled ki caseous pneumonia," or anatomically, 
" caseous hepatization." It might be thought very doubtful whether 
this change was in any way connected with the injections. Professor 
Virchow would himself have doubted this were it not that some of the 
cases showed it in a special degree, and he exhibited a portion of lung 
which had undergone caseous .hepatization to an extent the like of 
which he had not seen for years. The lungs were large, their lower 
lobes, especially the right, as voluminous as in ordinary hepatization ; 
but the foci were so thickly massed as to leave hardly any parenchyma 
between them. It was like a piece of black sausage richly laden with 
fat. The non-caseous parts were blackish and sharply demarcated 
from the caseous. The man was thirty-three years old ; he had had 
six injections, the last four weeks before death, and the injections 
were suspended because of supervention of continued fever and infil- 
tration of the lower lobes. This infiltration started just after the injec- 
tions, for previously there were only signs of induration at one apex, 
where was afterwards found an old indurated focus. In other cases, 
also, the condition of the lungs similarly differed from what is usually 
seen in phthisis ; in five of the cases there were more or less large 
masses of caseous hepatization, but not approaching the extent of the 
case related. 

They had lately a very remarkable case of a boy, three years of 
age, with tuberculosis of the vertebras and long bones, and where 
large tubercles were found in the brain. The injections had reached 
0.012 gr. It was a surgical case, in which spinal caries with psoas 
abscess and much disease of the joints and bones of the lower limbs 
existed. At the necropsy there were as many as seven so-called 
"solitary tubercles" — i. e., large caseous masses — in the brain and 
cerebellum ; but neither in them nor in their vicinity were any marked 
changes to be seen. There were a few spots of softening, but not of 
special import, and certainly none of the nodules showed any marked 
destruction." 

I wish I had space to quote the entire article, that I might 
not be accused of doing the distinguished author a seeming 
injustice, by selecting just such portions of it as suited 
my point, but if you will read it in The Lancet you will 
see that there has been no injustice done or advantage taken. 



THE BACTERIOLOGICAL WORLD. 251 

" It is imperative that the question should be fairly faced 
and impartially handled ; otherwise there may be witnessed as 
great a revulsion against the use of the ' remedy ' as there has 
been of indiscriminate enthusiasm at its first adoption/' In 
the first place Virchow claimed twenty-one post-mortems for 
himself, and a large number for his assistants in other hospi- 
tals and elsewhere. At first glance this looks rather serious 
for some five or six weeks' use of the lymph. But let us exam- 
ine or review the conditions of these cases and see if they are 
such cases as Koch said his lymph should be used in. Five of 
the cases Virchow eliminates himself after he has first accredited 
them to the use of the lymph, so we will only have the remain- 
ing sixteen. In these sixteen cases we have one boy, two years 
and a half old, with tubercular arachnitis, with some old 
caseo-pneumonic areas. One man, aged thirty, with old rectal 
fistula and numerous tubercular ulcers in colon. Five cases of 
more or less large masses of caseous hepatization. One boy, 
three years of age, with tuberculosis of the vertebrae and long 
bones, large tubercles in brain and psoas abscess. We are not 
told the condition of the remaining eight cases, but, if they were 
such as the ones described above, they were sorry cases indeed, 
and not one of them a fit subject for the lymph. The mighty 
have fallen very low when the great Virchow can stoop to 
make the use he did of such material. I take the following 
from the pen of Dr. 0. Isreal {London Lancet, December 6, 
1890), who was selected by Virchow to examine some specimens 
from patients operated upon after being treated with Koch's 
lymph. " It has been proved by Baumgarten that, in cases of 
progressive tubercular disease, the mere appearance of bacilli in 
the tissues of a certain part does not immediately induce pro- 
liferation of the tissues, which subsequently end in necrosis. 
On the contrary, a certain, though limited, multiplication of 
the bacilli in loco is a necessary condition for the occurrence of 
such necrosis." Now test by this fact, many of the conditions 
pointed out by Virchow as being probably due to the use of the 
lymph. Virchow says he performed twenty-seven post-mortems, 
and a large number by his assistants. A large number is a very 
indefinite term, but we will say that it means half as many, or 
ten, and we have a total of thirty-seven. In the British Medi- 



252 THE BACTERIOLOGICAL WORLD. 

cat Journal of March 14, 1891, you will find the following 
excellent tabulated report of all the cases receiving Koch's 
lymph, in the University clinics and " polikliniks" and other 
public institutions of Prussia. ( This tabulated report was 
made up from the report made by the order of Yon G-ossler, 
the Prussian minister of education. It only includes the 
cases treated in November and December). 



THE BACTERIOLOGICAL WORLD. 



253 



Table I. — Statistical Statement of the Action of Koch's Fluid in 
Tuberculosis of Internal Organs. 



DISEASES. 


Number of 
Cases 
Treated. 


T3 

0> 

U 

d 

O 

9 





1 
1 











s > 

■§ A 

■sa 
£~ 

72 
10 

1 


68 
10 

1 



7 
1 

1 



^3 

> 
o 

5h 

a 
i— i 

59 
6 

2 
1 

68 
11 

4 


31 
4 

2 
1 

158 
21 

8 
2 

23 
13 
3 

1 
3 



4 
2 


194 


T2 

> 

O 

B 
"3 

t> 

93 
13 

4 
6 

278 
37 

5 

7 

162 

45 

17 

7 

533 
95 

26 

20 

15 

12 

9 



1 

4 



4 

5 

14 

1 

586 


T3 


I. — PULMONAEY TUBERCULOSIS: 

1. Early pulmonary phthisis 

(a) With laryngeal tuberculosis. 

(b) With tuberculosis of other 

internal organs 


242 
30 

7 
8 

444 

85 

15 

14 

246 
60 

24 
16 








2. 


Moderately advanced pulmonary 



6 




(a) With laryngeal tuberculosis. 

(6) With tuberculosis of other 

internal organs 


2 




(c) With other diseases 


3 


3. 


Very advanced pulmonary phthisis 
(cavities) 


30 




(a) With laryngeal tuberculosis. 
(5) With tuberculosis of other 

(e) With other diseases 


5 

1 
5 


I.- 


—Pulmonary Tuberculosis (all 

grades taken together) 

(a) With laryngeal tuberculosis. 

(£>) With tuberculosis of other 

internal organs 


932 

175 

46 

38 

63 

45 

13 

1 

4 

14 

1 

4 

10 
18 

1 


10 

1 




1 



1 




1 








13 


147 
21 

3 

18 
16 



3 
1 




2 



171 


36 

7 

3 




(c) With other diseases 


7 


II. 
III.- 


—Laryngeal Tuberculosis 

With pulmonary tuberculosis. 
—Pleurisy 


4 
4 
() 


IV.- 


—Pernicious Anaemia 


1 


V.- 
VI.- 
VII.- 
VTTT 


—Tuberculous Meningitis 

—Peritoneal Tuberculosis 

—Intestinal Tuberculosis 

—Renal Tuberculosis 


2 
2 





IX.- 


—Urethral' and Vesical Tuber- 
culosis 


1 


X.- 
XI.- 


-Testicular Tuberculosis 

-Double Tuberculous Pyosal- 
pinx, with incipient pulmo- 
nary phthisis. 







Totals 


1061* 


46 



* The difference between the total number of cases and the num- 
ber in which the result of treatment is given is due to the fact that a 
few reporters give no information as to the result . 



254 



THE BACTERIOLOGICAL WORLD. 



Table II. — Statistical Statement of the Action in Koch's Fluid in 
External Tuberculosis. 



DISEASES. 


Number of 

Cases 

Treated. 


O 

5 



9 













1 












15 


1 ^ 

§ 2 

-1-3 S 

03 & 

78 

5 
51 

1 

3 


9 

1 
1 

2 


1 

1 



2 




148 


> 
O 

u 
S 

84 
6 

119 
4 

12 

1 

7 


4 
3 
3 
1 
1 



1 


2 
1 

237 


^5 

> 

O 
u 

a, 

s 

"3 

21 

3 
211 

22 

23 

7 
22 

7 
3 
1 

1 
1 
1 






14 

6 

298 


H3 

5 


I. — Lupus 


188 
27 

397 
48 
40 

10 

38 

8 
8 
4 
6 

2 
2 
3 

1 
1 

2 
16 

7 





With tuberculosis of internal 
organs 





II.— Tuberculosis of Single Bones 
and Joints 


6 


With tuberculosis of internal 
organs 


1 


III. — Tuberculosis of Several Bones 

and Joints 

With Tuberculosis of internal 
organs 


2 

3 


IV. — Tuberculosis of Lymph Glands. 

With tuberculosis of internal 

organs 






V.— Tuberculosis of Soft Parts 

VI. — Tuberculosis of Scars 

VII.-t-SCROFULODERMA 







VIII.— Lepra 





IX.— Rodent Ulcer 

X. — Tuberculous Anal Fistula 

XI. — Tuberculosis of Sheaths of 

Tendons 

XII — Scrofulous Eczema 




1 





XIII. — Scrofulous Keratitis of Both 
Eyes 





XIV. — Ear Diseases 





With pulmonary tuberculosis. 





Totals 


708 


q 







We see from this that, out of a total of nearly eighteen 
hundred cases of good and bad, we have only fifty-five deaths. 
It seems a remarkable fact that Virchow and his assistants 
should have performed over one-half the post-mortems of all 
the cases occurring in the entire Prussian kingdom. If this 
were in politics, instead of " poliklinicks," someone would have 
to rise to an explanation. 

There are many valuable lessons to be learned from the 
above tabulated report. The scientific world cannot look upon 
it and say that Koch's " tuberculine " is not a great boon, and 



THE BACTERIOLOGICAL WORLD. 255 

a great triumph in the sciences of the nineteenth century. 
We must remember that this report covers the first cases treated, 
which, of necessity, must be the most unfavorable. While in 
Berlin I saw some such cases that died, but they were in the 
very last stages of consumption, and I have seen here at home 
many such cases die just as soon without the lymph, and under 
the best of the old treatment. 

We should also remember that the climate of Germany at 
any season of the year is not favorable to the treatment of 
tuberculosis, and in the winter season, the time of these exper- 
iments, it is probably one of the most unsuitable of all climates. 
Furthermore, we all know that Germany has just experienced 
an extraordinarily severe winter. Nearly all, if not all, of these 
cases come from the poor classes and such, too, as are found in 
the large cities. While the hygienic regulations of the Prussian 
kingdom impressed me as being possibly as good as, if not 
better than, those of any other country, yet the foodstuffs of 
the poorer classes is not, I think, of such variety and quality as 
to enable them to resist the ravages of such a disease as tuber- 
culosis. Unless compelled by circumstances, what physician 
would ever think of treating, or selecting such circumstances 
under which to treat, tuberculosis ? Have not the fatal experi- 
ences of the past taught us and our patients, if possible, to flee 
from such surroundings ? In an unfavorable climate, in the 
midst of a severe winter, with a poor class of patients, many of 
them suffering from a complication of diseases, and located in 
large hospitals and in crowded cities, the "lymph" has accom- 
plished results which should fill us with hope and faith in its 
final triumph. It may do us good just here to consider for a 
moment our condition before the discovery of the lymph. I 
will give it in the words of Dr. T. W. Hime, of England : 

" The magnitude of the evils arising from the prevalence 
of tuberculosis was greatly increased by the terribly long dura- 
tion of the disease, which caused its economic importance to be 
so much greater than that of almost any other disease, and by 
the fact that adults, the most effective portion of the population, 
suffered so largely. It was cruel enough to see young children 
swept off rapidly by measles, whooping-cough and other dis- 
eases, but it was far worse to see their parents and adult 



256 THE BACTERIOLOGICAL WORLD. 

brothers and sisters slowly gnawed away by the relentless bacillus 
of tuberculosis. The modern discovery that the tuberculous 
patient is a source of infection to those around him, during the 
long, weary years of his wasting disease, certainly has not 
robbed it of any of its terrors, although the discovery, no doubt, 
will ultimately lead to precautionary measures and protection 
for those exposed to the risk. Up to the present time it must 
be admitted that practically we had been helpless against 
tuberculosis. Medical science had devised little more than 
uncertain methods of giving general tone and vigor to the 
tissues of the patient by good food, air, tonics, exercise, etc., 
so as to assist it in resisting the inroads of the organism of the 
disease. It is unnecessary to particularize the extent of the 
success attained. But certainly no scientific method of contend- 
ing against the disease had been even plausibly suggested until 
the present. Indeed, only a few years ago a distinguished 
authority gave a reason for the theoretical impossibility of 
devising a ' protective ' against tuberculosis — viz., the fact that 
one outbreak of tubercular disease does not protect against a 
second or further one. Suddenly, it may be said, all theories 
and opinions on this subject have been subjected to a rude 
shock, and in a most unexpected manner. The excitement and 
enthusiasm aroused by the announcement that Koch had dis- 
covered a successful method of dealing with tuberculosis have 
surpassed all similar occurrences. Within a few days of the 
announcement of the discovery thousands of medical men, 
including many of the most distinguished in every land, had 
abandoned their professorial and private work, and in the 
depth of winter were pilgrimaging to Berlin to see with their 
own eyes the new method in operation, and with a vague hope 
of bringing back some of the liquid for use at home." 

The opponents of the lymph talk of dangers, bad results 
and death from the use of the lymph. Does the tabulated state- 
ment of the first two months tell a story of danger, bad results 
and death ? But what is the story of tuberculosis without the 
lymph ? Could any story be more pregnant with dangers, bad 
results, to finally end in death ? Twenty-eight cures, 319 sub- 
stantial improvements, 431 improvements, or a total of 778 
cured or improved, out of 1,769 cases. Or, in the city of Berlin, 



THE BACTERIOLOGICAL WORLD. 257 

36 cured, 306 more or less improved, or a total of 342 cured, 
or improved, out of 623 cases. Is this a story of dangers, bad 
results and death ? To fully appreciate these results, we must 
not for a moment forget that the large majority of these cases 
were not such as Koch claimed would be benefited by his 
lymph. It was all right and proper that these cases should 
have been treated, and the results reported, but, in making up 
our verdict, let us give them their proper value. Let us again 
refer to the statement of Koch, viz.: " These experiences lead 
me to suppose that phthisis in the beginning can be cured 101th 
certainty by this remedy." What are the results of the first two 
months of treatment of 242 cases of early pulmonary tubercu- 
losis ? Nine cures, 72 substantially improved, 59 improved, 93 
unimproved. You may say that there is small encouragement 
m only nine cures. But remember this is only after two 
months' treatment. Who can say how many of the 72 or 59 
may not be transferred to the cured list, after longer treat- 
ment, and especially if they could be placed under more 
favorable climate and hygienic influences, and, last, but not 
least, when we have learned how to use the lympb. As our 
knowledge of diseases enlarges and becomes more detailed and 
accurate, our ideas of the stages of diseases are constantly 
changing. Have we not come to regard all of our infectious 
or contagious diseases as having a stage of incubation ? If I 
mistake not, these incubating stages are not characterized by 
any appreciable anatomical or structural lesion, and frequently 
with little or no constitutional disturbance. May this not be 
true of tuberculosis ? You may say, that, if this be true, the 
lymph will only act upon tuberculous material, the presence of 
which indicates that the disease is beyond the stage of incu- 
bation. Theoretically and anatomically, this is true, but, 
practically, it is not so. That is a hard saying, and hard to 
believe, but let us see. Many men and women are going about 
their daily work with a deposit of tubercle in their lungs, and 
they have never suspected it. They eat well, they sleep well, 
and work well, and all the functions of the body work well. 
Let me illustrate the point by a case which occurred at the 
Victoria Hospital for Consumptives, London, England. 



258 THE BACTERIOLOGICAL WORLD. 

One day, while visiting this hospital, Dr. Heron, who is 
one of the leading practitioners of London, and who has charge 
of this hospital, showed me a young woman whose case had 
been a little amusing, as well as very valuable in showing the 
diagnostic value of the lymph. Dr. Heron said he had selected 
this case to show that the lymph in a certain dose would have 
no effect upon a perfectly healthy patient. I think this patient 
had not only been examined by Dr. Heron, but by several 
others, and she gave not the slightest evidence of phthisis, nor 
could she give any family history of phthisis, and she had been 
healthy and strong all her life. A few hours after the injection, 
this patient had a strong reaction, and the local reaction in the 
lung revealed the spot. Under a few weeks'* use of the lymph 
this patient made a perfect recovery, and looked to me to be in 
perfect health, and she stated that she never felt better in her 
life. 

Such a case as this may be considered rare, but let us take 
another case that is not rare. A patient comes to you and he gives 
you a history of his case, and you suspect tubercle, but he tells 
you that, up to a short time ago, he was in perfect health ; 
never had to consult a doctor before, and tells you the amount 
of work he could do„ and what he could stand. You examine 
his lungs, and you find they are all right, as far as you can 
judge, and you relieve his mind and apprehensions by telling 
him there is nothing the matter with his lungs, but to be 
careful, and you give him a tonic, and he gets better. It is 
not long before he comes back, and the same examination and 
opinion and advice 1 are given him as before ; but, after a varied 
length of time, you discover what we call the signs of incipient 
phthisis, and, if he is in good circumstances, or can afford it, 
we advise him to go to some climate like Florida, or our 
western slope. He does so, and in a few short years, as a 
rule, he returns, that his bones may have a resting place among 
his kindred at home. I believe these patients suffer from, or 
present, incipient phthisis some time before the most skilled 
diagnostician can detect it. I believe, by the use of the micro- 
scope and the lymph, we shall be able to make a diagnosis in 
time to save most of these cases. In some future issue of this 



THE BACTEKIOLOGICAL WOKLD. 259 

journal I may make some suggestions as to the therapeutical 
applications of the lymph. 

Let me close this lengthy article by saying that I believe 
we have ever reason to be encouraged in regard to the final 
success of the lymph. We should congratulate ourselves and 
suffering humanity that Koch says that he is not through with 
it, and is still working upon it. I take great encouragement 
from Koch's attitude during the test of his lymph. He gave it 
into the hands of friends and foes alike, and, although in the 
past half year, he has passed through an experience that has 
been the most remarkable of any living man, yet he has con- 
tinually borne himself with a quiet placid dignity that is born 
only of the consciousness of final triumph. As a fitting close 
to this article let me insert the following from The London 
Lancet of January 10, 1891 : 

Koch came to my institute on April 30, and I can truly boast that 
in the first hour of our intercourse I recognized in him an unequaled 
master of scientific investigation ; his method, proceeding with rigid 
consistency from step to step, the elegance and certainty of his experi- 
ments, the classic clearness of his statements, were all as perfect in 
his first, then just completed, work on anthrax as in all his later 
researches. For Koch's works are distinguished from those of most 
investigators by the circumstance that he does not publish them till 
they are finished to the last point. Others cart up stones to be used in 
the building up of science, or draw a new plan or add a new wing, a 
new story, a new roof, but they finish only the brickwork, and leave 
it to others to complete the building and make it habitable. Koch, on 
the other hand, does not let his scientific fabrics leave his hands till he 
has made them completely fit in all their details as well as in the mam 
for the use of others, who have then nothing more to do than to add 
this or the other little furnishing. All Koch's works have been so 
complete in form and contents that nothing remained for those that 
followed but to confirm them, for it was not possible to add anything 
essential. Such were the first works of 1875 on anthrax, that, on 
wound infection, the numerous treatises in the communications of the 
Imperial Office of Health, the magnificent researches on the tubercle 
and cholera bacilli, and such beyond doubt his latest discovery will 
prove. 



260 THE BACTERIOLOGICAL WORLD. 

OBSERVATION ON COUNTRY SURGERY. 



BY PROF. A. W. MCALESTER, M. D. 

Can aseptic and antiseptic surgery be practiced outside 
of well regulated hospitals, especially in small towns and 
country places ? I answer, yes. Since we understand the 
science of aseptic and antiseptic surgery, the advantage is 
decidedly in favor of country air. There are fewer pathogenic 
bacteria in a sparsely inhabited region than in a dense city; 
fewer persons to furnish effete material. 

There is an inherent power to resist disease. The human 
system might throw off a few pathogenic bacteria but would 
succumb to larger numbers. The country physician has really 
less use for antiseptics. Aseptic surgery is much more desira- 
ble than antiseptic surgery, for the reason that the antiseptics 
irritate cells. The less a cell can be irritated the quicker it 
can repair the injured tissues. Hence asepsis is decidedly 
preferable from a scientific point of view, and, from a 
practical standpoint, much easier applied. Really less skill 
upon the part of the practitioner is required. The keynote of 
asepsis is "heat." Everything that comes in contact with your 
patient must be heated. 

To prepare the room : — 

The furniture should be removed, the carpet taken up, the 
floor scrubbed, the windows washed with hot water and dried 
with heated air, as hot as possible. The hotter the air the 
better, as heat modifies all pathogenic germs. Use plain furni- 
ture because it can be more thoroughly cleaned. See that 
every article of clothing has been subjected to at least 212° 
moist or dry heat. 

Now for the day of operation : — 

See that your patient has a hot bath and no clothes on 
but clean underclothing. See that yourself and attendants are 
cleaned. ( Use a nailbrush.) Eender every instrument aseptic 
by hot water, and either roll them in aseptic cloths or put in an 
aseptic dish. Cool all water used, in a tightly covered vessel ; 
never cool by pouring cold water into your vessel. It had as 
well never boiled, as a "little leaven leaveneth the lump." 
Drainage tubes are hard to render aseptic. 



THE BACTERIOLOGICAL WORLD. 261 

G-ive the wound a final washing ; close it, and sprinkle 
the edges with iodoform or its equivalent, then cover with 
either aseptic rubber tissue or cloth, and cover well with, 
sterilized absorbent cotton, I gathered this use of cotton from 
Dr. P. Paquin, who kept some broth eighteen months in a flask 
using only sterilized cotton for a stopper. The (i soup "was 
as good at the end of eighteen months as it was the day it was 
put there. The air could pass through the cotton, but no 
putrefaction, thus showing that the cotton filtered out the 
putrefying germs. If it will do its work in a flask why not on 
the human subject ? The maxim is, no bacteria, no decay. 

Now for the second dressing : — 

So long as there is no fever or odor do not redress till the 
day comes for removing the stitches. Then again have every- 
thing rendered aseptic that touches the wound. As a rule 
antiseptics will be used now on account of their greater con- 
venience in being applied. There is less objection now to 
antiseptics as you have a skin covering, and your bichloride 
of mercury will not irritate the cells of the part so much. 

Pyoktanine seems to be coming into use of late, because 
it does not provoke irritation, and is a true germicide. The 
stains left upon the surgeon *s fingers are often objectionable, 
but alcohol will remove it, if applied before washing. 

Where is there a man who cannot be clean ? If aseptic, 
he can dress any wound ; if not aseptic, soak everything used in 
an antiseptic. 



262 . THE BACTERIOLOGICAL WORLD. 

SELECTIONS. 



THE FILTRATION OF WATER.— {Concluded.) 



BY M. E. DUCLAUX, 

Member of the French Institute, 

Paris, France. 

It is evident that, with this constitution, a sand filter is 
something very fragile, and it is clear, also, that it cannot pre- 
vent the passage of a few microbes with the water. The filter 
cannot, therefore, be perfect. We can reduce greatly the num- 
ber of bacteria in the filtered water by slackening the speed of 
filtration ; but then the filter works no longer in the industrial 
conditions. We may reduce the figure also, as much more as 
the water is less impure ; but the number given above, concerning 
Zurich, shows that, even with the relatively pure waters of the 
lake, the ratio of bacteria in the filtered water never falls to 0* 
Consequently, bacteria are constantly dragged out of the filter, 
and, when we reflect, we see that it could not be different. 

The sand filter is then a bad instrument, out of which 
engineers have learned to draw the best possible results. This 
manner of representing very costly installations has met first 
with opposition. Under the pretext that it was about an indif- 
ferent matter to allow to the filtered water very rich or very poor 
in bacteria, and that the water drawn therefrom contains 
always about the same number, we had reached the point of 
considering a sand filter as formed some way of two superposed 
systems ; a perfect filter, allowing the passage of scarcely any 
microbes, formed chiefly by the superior layers, and the nutri- 
tive media, formed chiefly by the inferior layers, where the 
multiplication of microbes takes place, as it occurs in conduit 
and distributing water pipes. 

This conception had the incontestable advantage, in the 
filtration and multiplication of microbes, of regarding the 
processes as two different phenomena, and to consider afterwards 
the germs found in the filtered water as independent of those 
met with in the water before filtration ; i. e., in a word, to 
quiet the mind on the use of an 'apparatus which replaced the 
dangerous microbic species of alimentary water by common 



THE BAOTEKIOLOGICAL WOELD. 263 

species. But the argument on which rested this conception 
was scarcely tenable. When a paper filter is perforated or 
weakened, in one or several points, the proportion of matter in 
suspension in the water passing through is not necessarily 
in proportion to the quantity in the water poured therein. This 
is what we often have occasion to observe in filtering, for 
instance, a liquid in which sulphate of baryta has *been precip- 
itated. The water of a Zurich filter may, likewise, carry the 
same quantity of germs that the water of a Spree filter may, 
without proving that the germs are from another source than 
the filtration water. 

Another argument in favor of the same conception of the 
filter was, that the microbic species of the filtered water are not 
the same as those in the water of filtration ; they are, in general, 
much less numerous. But this proves that there is cultivation 
in the filter, and that the species which it suits best develop the 
better ; it does not argue that there cannot be a simultaneous 
dragging of a few of the microbes by the water. 

This is, by the way, the recent experimental conclusion of 
Mr. 0. Fraenkel. In filtering through a ripe filter water loaded 
with a particular microbe easily recognized, the bacillus viola- 
ceus, he has observed colonies of this organism in filtered water. 
This experiment is the more probable that, to avoid the possible 
objection of gradual successive invasion of the filter by the bacilli, 
multiplying by the means of organic matters carried into the 
water by the culture media broths diluted therein, Mr. Fraenkel 
had taken the precaution to make this culture in a media orig- 
inally very diluted, the mixture of which with the water aug- 
mented its nutritive matter only in infinitesimal proportion. But 
then, if these microbes of the water penetrate the whole thickness 
of the filter, the question arises, why do they not develop them- 
selves equally throughout the height or body? It is not the 
organic matter that is wanting. We have seen that, in the 
Zurich water, judging from the amount of oxygen which it 
borrows from the hypermanganate of potash, its variation from 
top to bottom is very slight, and that there was left in the 
filtered water more than three-fourths of the quantity contained 
at its entry into the filter. It is true that oxygen disappears more 
rapidly, but all microbes do not need it, and on the other hand 



264 THE BACTERIOLOGICAL WOKLD. 

there remain some at the bottom. For instance, in water filtered 
through a ripe industrial filter we find about half the quantity 
contained therein before filtration. Why, then, does not the 
filter become more impregnated in all its thickness, and why is 
life much more abundant in the superior layers? 

This question is important, because if we solve it, we have 
a chance to find in the solution some light on the subject of 
this second question of the same nature : Why is it that, in the 
soil, microbes are in preference on the surface, and gradually 
diminish as we proceed deeper, until at a slight depth we find 
layers absolutely sterile? 

We see immediately that life is more abundant in the 
superior layers because there exists more oxygen and more 
organic matter, and, from the moment that the filter begins to 
ripen, because of the greater renewing of germs. We see, on the 
other hand, that a layer so lively on the surface can only render 
life more miserable in the interior. It is the fate of all microbes 
to spoil their media of culture for themselves and [ sometimes* ] 
others, either in depositing detrimental substances acting in 
the manner of antiseptics, or in arresting therein useful matters. 

[the end.] 



BACTERIA— A PRACTICAL STUDY IN BIOLOGY. 



BY V. A. LATHAM, F. R. M. S., F. S. S., ETC. 
DEFINITION. 

Biology is the science of living things : — fiioS — life, XoyoS, 
science. It may be divided into two branches, each of which 
corresponds to a distinctive mark of living things. They may 
be recognized, first, by their form — morphology ; second, by 
their effects ( movements and mutual actions, chemical ), 
physiology. The simplest possible organism is torula, the 
single cell of saccharomyces, or yeast. There is no well-defined 
limit between ferments and fungi, or, again, between fungi 

* The translator. 



THE BACTERIOLOGICAL WORLD. 265 

and bacteria. Their smaller size is the principal difference 
which separates bacteria from ferments, since in other respects 
these two classes are for the most part alike in form and 
organization. Everyone now speaks of microbes, yet few of 
those who make use of the term have any clear conception of 
the organisms in question, or could give an exact account of the 
function which microbes fulfill in nature. And yet this func- 
tion concerns us all. There is much to be done before modern 
society is practically on a level with the achievements of science ; 
many prejudices must be uprooted and many false notions 
must be replaced by those which are sounder and more just. 

In order to indicate the organisms which produce diseases, 
the English and Germans use the word bacteria, which is only 
the name of one of the peculiar species assigned to this group, 
and the one with which we have been longest acquainted. In 
this case the name is generalized and applied to the entire 
group. The Italian authors, who have been recently occupied 
with the study of microbes, have, on their part, adopted the 
name, protista, proposed by Haeckel, and of which the sense, 
although not the etymology, is almost the same as that of the 
word, microbe, which is the Erench term for bacteria, and only 
signifies a small living being. It decides nothing as to the 
animal or vegetable nature of the organisms in question. 
Bechamp terms microbes, microzyma, or small ferments, since 
the chemical reactions which result from their vital activity 
are generally ferments. Bacteria are distinguished from 
animal cells by being able to derive their nitrogen from 
ammonia compounds, and they differ from the higher vegetable 
cells in being unable to split up carbonic acid into its elements, 
owing to the absence of chlorophyl. 

The schizomycetes, or "splitting-fungi" — German, spalt- 
pilze ( (Tjz^Gtf, I split ; juvfcoZ, fungus) — are unicellular plants, 
which multiply by repeated subdivisions in one, two or three 
dimensions of space, and also frequently reproduce themselves 
by spores, which are formed endogenously. They live either 
isolated or combined in various ways, in fluids and in living or 
dead organisms, in which they produce decompositions and 
fermentations, but never alcoholic fermentation. 



266 THE BACTERIOLOGICAL WORLD. 

I shall use here the method I gave in my paper on ammbce 
in the American Monthly Microscopical Journal for July, 1889, 
which may be altered to suit requirements : ( a ) Structure, 
(Z>) digestion, (c) absorption, (d) circulation, (e) respiration, 
(/) secretion, (g) nervous system, ( h) sense organs, (t) motor 
organs, (&) reproduction, (I) development, (m) classification. 

PRACTICAL WORK. 

Every plant or animal must be seen, examined, dissected 
and drawn. 

(a) Structure. 

1. Occurrence. — Wherever there is putrefaction they 
are present in vast numbers. In water, in soil, in sewage, in 
the intestines and in uncleanly persons — especially on the skin 
and between the teeth — various species may be found. The 
red color noticed in mouldy bread is due to bacteria. Such 
diseases as diphtheria, typhus fever, consumption, cholera, 
anthrax ( or l ' wool-sorters' disease " ), appear to depend on 
their presence. 

Many cases of fermentation are due to the presence of 
these organisms, and the putrefaction of plants and animals 
seems to result from fermentations originated by them. Stu- 
dents who may wish to prepare these organisms without the 
help of disease will find it best to infuse hay in warm water for 
thirty minutes ; filter, and keep the filtrate in a warm place. 
Note the changes which go on in it. At first, clear ; in twenty- 
four to thirty-six hours, it becomes turbid ; later on, a scum 
forms on the surface, and the infusion acquires a putrefactive 
odor. One very common bacterium may be easily procured, — 
bacterium termo, Dujardin ( " zoophyt/' p. 212 ) ; ( synonyms, 
monas termo, Mueller ; (?) palmella infusionum, Ehrenberg ; 
zooglma termo, by Cohn ) — by taking half-a-glass of ordinary 
water from a spring or river, and leaving it for some days on a 
table, or chimney-piece, the vessel being uncovered to allow 
free access of air. The water from the top of a small aquarium 
will also show them, or a small piece of meat kept in water at a 
moderate temperature. 

2. Size. — They are very minute. Hence arises a great 
danger of the confusion of other bodies with these on the 



THE BACTERIOLOGICAL WORLD. 267 

part of the student. The cells are shortly cylindrical, oblong, 
about 1.5 to 2/i long, with a flagellum at each end. 

3. General structure. — They are masses of protoplasm 
of various shapes, that are, in many cases, not surrounded by a 
distinct wall. Protoplasm is derived from 7tf)GQto$, first ; 
nXafffAa, formative matter ; i. e., matter that can be shaped 
into other substances, such as the tissues of a plant or of an 
animal body. It is semi-fluid, granular in appearance, and 
consists of four elements — carbon, hydrogen, oxygen and nitro- 
gen. It has the power of contractility. The protoplasm 
adheres to iodine, and it becomes brown. If the piece of 
protoplasm is without an enveloping wall, as in many bacteria, 
and consists of protoplasm, and protoplasm only, we have 
a plastid. If the protoplasm is invested with a distinct wall, 
that is, wholly distinct of structure, the being is a cytode ; for a 
cytode is a free mass of protoplasm, enveloped by a distinct 
wall. This wall is more transparent than the contents. In 
the compound form it is only to be seen just where the joints, 
making up the compound form, come in contact, one with the 
other. In the still condition of these organisms, the wall is 
exceedingly gelatinous. This still stage, with its thick envelope 
of jelly-like material, is known as the zoogloea stage ( Zgoov, 
life ; yAoioS, viscid ). 

4, Different forms of bacteria are classed under this 
heading. When they occur alone, they may be either spherical, 
as in certain forms of micrococcus, or rod-like, as in the true 
bacteria. When they occur aggregated together, they may be 
of the form of rudimentary cells, as in other varieties of micro- 
coccus, or as rod-like structures, joined together. Even this 
last arrangement admits of subdivision, as rod-like parts may be 
conjoined end on end, or at an angle (not of 180°), or wound 
in a spiral. The following table may make this clear : 

t i i a \ Spherical — e. a., Micrococcus. 
Isolated < t> -, V1 -da • 

( Kod-like — e. g., Bacterium. 

rg , Spherical — e. g., Micrococcus. 
Bactebia .i.-§ \ • f Pfrts joined end on en(]^. Bacillus 

a J tt it at an angle — e.g. Vibrio 

h* ' Rod-like \ (C " spirally ( often a double 

spiral) — e. g., Spirillum 
and Spirochete. 



268 THE BACTERIOLOGICAL WORLD. 

5. Effects of reagents and temperature: — 

1. Iodine stains all formative matter or protoplasm 
brown, and occurs with all formative matter, ready to be, but 
not yet actually, transformed into a plant or animal substance. 
The outer wall does not stain. 

2. Magenta or carmine solution also stains protoplasm. 

3. A temperature of o°0. (32°F.) and of 60°C. (140°F.) 
kills them. These are their limits of temperature. They 
thrive best at 30° 0. (86° F.). 

4. Desiccation: They can survive a lengthy period of 
drying or desiccation, and on adding moisture they revive, 

( i ) Digestion. — Bacteria feed on decaying organic mat- 
ter. Hence their name of saprophytes ( Ga-rrpoS — putrid, 
<Pvrov — plant). Pasteur's fluid causes them to thrive. This 
food is taken in by the whole of the general surface. There is 
no definite mouth or digestive apparatus. 

(c) Absorption. — As in many of the lower living 
beings, the function of absorption is merged with that of diges- 
tion. The whole surface of the bacteria absorbs food. But 
that very absorption is here in reality a digestive process, and 
results in the in-taking of food. 

( d ) Circulation. — Nothing to be noted. 

( e) Respiration. — The organs of respiration are on the 
whole body surface. The lower we descend in our study of 
biology the more do we find every function performed indiffer- 
ently by every part. Nothing is specialized. The whole body 
oreathes, reproduces. 

(f) Secretion. ) mi • i.l- j. i 

(g) Nervous system. ./^"if 1 ?] be 
(A) Sense-organs, j studied under these heads. 

( i ) Motor organs. — I would here warn the student 
against a movement noticeable in these organisms that he will 
be apt to regard as due to life. It is highly important not to 
confound the two kinds of motion. That of dead matter is 
merely mechanical, not vital, and resembles somewhat the 
swinging motion of a pendulum, or' that of "a floating buoy 
round its mooring." This movement can be seen whenever any 
very small particles of matter, living or dead, are suspended in 
a liquid. Very finely-divided charcoal, camphor, cinnabar or 



THE BACTERIOLOGICAL WORLD. 269 

gamboge, if placed in water, will exhibit similar movements. 
It is called the Brownian movement, after Brown, who thor- 
oughly investigated it. It is simply a molecular movement, 
due to mechanical, not to vital, causes. Some of the bacteria 
only show this motion, and are then said to be in the still stage. 
As in this condition they are usually surrounded by a quantity 
of jelly-like material, this stage is known as the zooglcea stage. 
Very often these organisms are in an actively mobile condition, 
and undergo movements of translation from place to place. 
The cause of these movements is in most cases not known. It 
is possible they may result from the general contractility of the 
protoplasm. But, in spirillum volutans, at each end a cilium 
or long, fine, hair-like process has been seen, and by the move- 
ment of the cilium that of the spirillum as a whole is effected. 
Vibrio has a wriggling motion, which is really due to its having 
a zigzag arrangement of its joints, and also a rotation upon its 
own axis. Bacillus is always free-swimming. 

( h ) Eeproduction. — This in living things is either 
asexual or sexual. The former is called agamogenesis {a — with- 
out, ya/AoS — marriage, yevsaiS — reproduction ). The latter, 
gamogenesis. Only the asexual or agamogenetic is known in 
connection with those low forms of living things. The partic- 
ular method encountered here is division or, technically, fission 
( findo — I split ). Certain divisions of the bacterium undergo 
transverse division. A cytode is divided into two by an 
ingrowth of its wall, and the cytodes thus formed last a longer 
time, and can resist injurious agencies more effectually than 
their fellows. By such division a bacterium becomes two bac_ 
teria. Bacillus also produces spores, e.#., rounded structures that 
are thrown off from the parent and developed into a new being. 

(/) Development. — Nothing can be said here, as the 
new individual is of the same nature as the parent. 

(m) Classification. — Cohn classifies the bacteria 
according to their form into sphcerobacteria ( globular cells ), 
microbacteria ( minute, rod-like cells, bacteria proper ), desmo- 
bactema ( large rod-like or filiform cells, filobacteria ) and 
spirobacterja ( twisted or spiral cells ). The student should 
rigidly adhere to order in this. For our convenience we divide 
everything in the universe as mineral, animal and vegetable. 



270 THE BACTERIOLOGICAL WORLD. 

We regard each kingdom as divided into sub-kingdoms. These, 
again, contain classes. Each class is made up of orders. An 
order has genera (genus — kind). A genus consists of species. 
Hence, in classifying bacteria, we say its food is inorganic; 
therefore, it belongs to the kingdom vegetabilia. It is a flower- 
less plant, without stamens and carpels ; therefore, it belongs 
to the sub -kingdom cryptogamia (fcpynroS — hidden, yajjoS — 
marriage ) . There is no distinction of axis and appendages, as 
in the stem and leaves of a tree ; therefore, it is of the group 
thallophyta ( OaWoS — a sprout, cpvrov — a plant ). It is with- 
out green coloring matter, chlorophyl ; therefore, of the class 
fungi. Gamogenesis, unknown, fission the only form of agamo- 
genesis ; therefore, of the order schizomycetes ( cjz^cy — I split, 
fxvfcoi — fungus ). Genus, bacterium, vibrio or bacillus, etc. 

For the chemical composition of bacteria, we are indebted 
to Nencki. Their constituents vary slightly according as to 
whether the bacteria are in zooglcea, or in the active state. In 
100 parts of the dried constituents, there are the following : 

A nitrogenous body, 84.20; fat, 6.04; ash, 4.72 ; unde- 
termined substances, 5.04. This nitrogenous body is mycopro- 
tein, so called, and consists of carbon, 52.32 ; hydrogen, 7.55 ; 
nitrogen, 14.75, but no sulphur or phosphorus. The nitro- 
genous body appears to vary with the species, for in bacillus 
anthracis a substance has been obtained which does not give the 
reactions of mycoprotein, and, therefore, is distinguished as 
anthrax-protein. Bacteria can and do utilize the last traces of 
energy in urea. It is the bacteria which play the most import- 
ant part in disease, exciting both general and systemic 
affections. The others, e. g., blastomycetes (yeasts), and liypho 
mycetes (moulds) — exert a merely local influence. The animal 
parasites become dangerous in virtue of their size, or multitude, 
or by penetrating into vital organs. Special reagents or stain- 
ing processes have to be employed to discover them ; sometimes 
certainty is only reached by experimental cultivation of the 
products of disintegration of the tissue in question. 

Eeagents. — Water is essential for their growth, though 
deprivation does not kill all the bacteria. Different species 
grow best on different nutrient media. 



THE BACTERIOLOGICAL WORLD. 271 

Effect of temperature. — Here again they vary, many 
grow best at the temperature of the blood, and hence the value 
of agar-agar media, which is not liquefied at 37° 0. (or 98.60 F.). 
The B. tuberculosis will only grow at a temperature varying 
between 30° C. (86° F.) and 41° C. ( 109° F.). 

Cold: — The bacteria seem to have a special power of 
resisting cold, even comma bacilli, if exposed to a temperature 
of — 10° for an hour, and bacilli of anthrax after exposure to a 
temperature — 110o C, still retain their vitality. The spores 
retain their vitality after immersion in boiling water, but are 
destroyed by prolonged boiling. 

Movement prevents their growth. 

Gases. — Hydrogen and carbonic acid are believed to stop 
movement of the motile bacteria. Chloroform is believed to 
arrest the changes brought about by the zymogenetic species. 

Electricity. — Oohn and Mendelssohn found that a con- 
stant galvanic current produced a deleterious effect owing to 
electrolysis. 

Light. — Sunlight is fatal to putrefactive bacteria. 

Carbonic acid. — A 5-per-cent. will kill the spores of 
B. anthracis in 24 hours. Three per cent, will not do so in the 
same time. A 1-per-cent. kills the bacilli. 

Chloride of zinc in 5 per cent, has no effect on anthrax 
spores, even when they ha^ve lain in it for a month. 

Sulphurous acid is not a good disinfectant, and will not 
penetrate compact masses or bundles. 

Corrosive sublimate has the most powerful effect on 
the organisms, an aqueous solution of 1 : 20,000 kills the spores 
of bacilli in 10 minutes. A solution of 1 : 5,000 is thus a cer- 
tain disinfectant. 

Iodine, bromine and chlorine are far more active 
than sulphurous acid. Bacilli cease to grow in iodine of 
1 : 5,000, and of bromine of 1 : 1,500. 

The oils of thymol, cloves, peppermint, mustard, tur- 
pentine, eucalyptus, all restrain their development ; the latter 
being very good, and is especially useful in dental cases to pre- 
vent bacterial growth in the teeth, as it penetrates the tubules. 
All disinfecting reagents should be used in aqueous solutions. 



272 THE BACTERIOLOGICAL WORLD. 

PRACTICAL WORK. 

First. — Observe Brownian movement in gamboge, etc., 
suspended in water. 

Second. — Turbid hay infusion under high power. 

Third. — Stain the various organisms. 

Fourth. — Stain with magenta. 

Fifth. — Observe cilia in spirillum. 

To stain the bacteria. — The following methods are 
useful for all bacteria, micrococci, and bacilli, except those 
of tubercle and leprosy : Dry a film of fluid containing the 
bacteria on a cover-glass, pour a little staining solution into a 
watch-glass, and place the cover on it, with the dried material 
downwards, leave for from 5 to 60 minutes. The time can 
only be learnt by practice, as the materials differ. When 
thoroughly stained, wash the cover-glass in methylated or recti- 
fied spirits until all the superfluous color is removed, taking 
care that it is not washed too much, or the bacteria film 
washed off. Then dry the cover-glass by holding it on its 
edge on blotting-paper, allow to dry. Then mount in Canada 
Balsam. 

TO STAIN SECTIONS OF TISSUE CONTAINING BACTERIA. — 

Place them in the stains given below and leave for some hours. 
When deeply stained, wash in water to remove excess of stain, 
then lay them out flat in spirit and leave till no more color 
comes away. Transfer to absolute alcohol which fixes the 
color, then to clove oil, and mount in Canada Balsam. 

The best stains are Gentian Violet, Spiller's Purple and 
Methyl Blue, the great difficulty being to obtain good samples 
of the dye, as the manufacturers change the numbers often, 
and the anilins differ greatly. 

Gentian violet (2 percent.) — Take 2 grms. of the powder, 
rub it up in a mortar with 10 cc. alcohol (sp. gr., 0.830), 
some prefer that in which 2 cc. of anilin oil has been dissolved. 
Then add gradually 99 cc. of distilled water while stirring. 

Spiller's purple. — Make as above, using the same 
strength. 

Methyl blue. — A 2-per cent, solution in distilled water 
and a little alcohol added whilst triturating the powder, previ- 



THE BACTERIOLOGICAL WORLD. 273- 

ous to the water being added. ( A little Liq. Potasses, 1 cc 5 
may be added in preference to alcohol if desired.) 

Gram's method, with a saturated alcoholic solution of 
gentian violet and anilin water and iodine of potassium, is 
good. 

For bacilli ik sputum and tissue. — The methods are 
very numerous, but I prefer Gi'bbes* Double-Stain or his 
Magenta and Nitric Acid methods, as they are always to be 
relied on if the dyes are pure and take very little time. I have 
seen and prepared specimens of sputa in five minutes for diag- 
nostic purposes. The bacilli do not fade. 

The ziehl-neelsen method and Sulphuric Acid is also 
very similar to the above Magenta, but I think the Magenta is 
to be preferred. All the other methods can be found in the 
various journals and tried by those who wish to experiment ; 
but the two stains given are all that are required by the stu- 
dent and busy practitioner. — The Journal of Microscopy and 
Natural Science* 



FISTULA IN ANO IN PHTHISIS. 

BY HEXEAGE GIBBES, M. D. 
Professor of Pathology in the University of Michigan. 

Anal fistulae are common in phthisical patients, and it was 
formerly thought that they exercised some protecting influence 
against this disease. The belief was common among practi- 
tioners that a patient with* suppurating rectal fistulae was, if 
not altogether prevented from acquiring the disease, at any rate 
when suffering from phthisis, enabled to resist the advance 
of the disease as long as the fistula remained open. Experience 
has, however, taught us that getting rid of a suppurating cavity 
is decidedly beneficial to the patient. 

I wish to point out what seems to me to be a possible cause 
of fistula occurring in cases of phthisis. * * * 

This sinus is about one-eighth of an inch in depth, it 
widens out at the bottom, and in the anterior wall, there is a 
lymph follicle, which is in close contact with the epidermis. 



274: THE BACTERIOLOGICAL WORLD, 

Directly under the epithelium of the anterior wall, and in 
the angle formed by the epidermis as it passes in towards the 
mucous membrane of the rectum, is the internal sphincter. 

The structure of the parts then is this, between the external 
and striped muscle and the internal non-striped muscle sphinc- 
ters there is a deep sinus having at the bottom in the anterior 
well a mass of lymphoid tissue. 

This resembles an ordinary solitary gland, the central por- 
tion is composed of dense adenoid tissue, and is partially separ- 
ated from the surrounding diffuse adenoid tissue by a lymph 
sinus, the walls of this sinus being formed of a fenestrated 
nucleated membrane. It resembles a tonsil in structure and 
arrangement. 

It is now a well-known fact from experimental investiga- 
tion that the tissues first affected after inoculation with 
phthisical material are those of the disseminated lymphoid 
follicles in the lungs, spleen and other parts ; it is, therefore, 
perfectly justifiable to conclude that in a case of general 
tuberculosis this lymph follicle at the bottom of this sinus may 
become the seat of tubercular change followed by breaking 
down and subsequent ulceration. This ulcerative process 
would have to extend only a short distance internally to involve 
a so-called sweat gland, the duct of which passes upwards 
through the anterior fibres of the internal sphincter ; and there 
opens on the surface. In this way there is direct communica- 
tion with the mucous membrane about half an inch or less 
inside the external sphincter. It must be remembered, how- 
ever, that on the introduction of # the finger or a speculum the 
sinus before mentioned would be spread out and obliterated, 
and this will account for the statement made by many sur- 
geons, that the internal opening is found much higher up. 
The same fact will account for the statement that the internal 
sphincter embraces the lower one and one-half inches of the 
rectum, as a matter of fact which can be verified by an exami- 
nation of a section through the parts. Quain's description is 
perfectly correct, namely, that the internal sphincter embraces 
about the lower half inch of the rectum. 

Some cases of fistula in ano are described in which the 
external opening exists outside the external sphincter. These 



THE BACTERIOLOGICAL WORLD. 275 

cases may be accounted for by the ulceration extending out- 
ward until it reaches the body of some of the numerous glands 
which lie outside the external sphincter, with their duct lead- 
ing directly to the surface. 

In this way a fistula may be formed, with its internal open- 
ing within an inch inside the anus, and having the external 
opening either at the bottom of the sinus, or, if the ulceration 
has extended further, it may be outside the external sphincter. 

So far as I know, these facts have never been mentioned 
before in any works on surgery or anatomy, and may prove of 
interest. 

ANTINOMYCOSIS BOVIS. 

I have lately made some observations on a case of actin- 
omycosis in the tongue of a cow, more especially with a view to 
determining the growth and arrangement of the mycelium. 
The published accounts vary -very much in their descriptions, 
and sDme observers seem to have failed to find the mycelium at 
all. On reading the different accounts and comparing the 
illustrations given, I find that either the ray itself must vary 
very much in size in different cases, or the measurements given 
are incorrect; also the manner in which it stains must vary 
according to whether it is in a fresh or hardened condition. 

In sections of the hardened organ I find it very difficult to 
stain the ray itself, and altogether impossible to stain the 
mycelium. This may of course be the result of the hardening 
method. I find the rays in this case, as well as in some others 
I examined formerly, to be much smaller than generally repre- 
sented, and to be rather infrequent in their occurrence. 

On the other hand, I find the mycelium permeating all the 
connective tissue under the superficial epithelium of the tongue, 
and in some parts even penetrating it. Wherever the mycelium 
abounds, there the section has a pale yellow appearance, except 
in those parts where it has set up an inflammatory action. 
Here there is typical inflammatory infiltration, and the leuco- 
cytes stain deeply. In only some of the inflammatory foci can 
rays be found, but this may be from the sections containing 
only a peripheral portion of an inflammatory nodule produced 



276 



THE BACTERIOLOGICAL WORLD. 



by the presence of a ray. It seems to me that the mycelium 
does not set up much irritation, but that the rays invariably do, 
and in many sections I have found the inflammatory action has 
gone on until disintegration is produced, and a minute abscess 
formed. The growth is not confined to the subepithelial tissue, 
but in places has penetrated into the epithelium and caused 





Portion of the Tongue of a Cow X 130. 
Longitudinal Section. 



inflammatory action and breaking down there. 

I have also noticed a peculiar feature in the growth of this 
fungus, that is, the manner in which it penetrates a muscle 
fibre and increases in its interior. 



THE BACTERIOLOGICAL WORLD. 



277 



The two accompanying photographs of this condition will 
fully explain it. They were taken with one of E. & J. Beck's 
t 4 q- in. magnification 130 diameters. In the one a muscle fibre 
is cut longitudinally, and has the mycelium occupying its 
centre ; the other shows a fibre cut transversely. The appear- 
ances presented by these fibres are exactly similar to what have 
been described as Bainey's or Miescher's corpuscles. 

Ziegler, volume 1, page 346 ( English edition ), says : 
" Our knowledge of the organisms known as 'Miescher's 
corpuscles' or 'Rainey's corpuscles' is still very defective. 
They are cylindrical or tube-like bodies, found not infrequently 
in the muscles of the pig, ox, sheep and mouse. They con- 
tain an innumerable multitude of small oval and reniform 
corpuscles." 




Portion of the Tongue of a Cow X 130. 
Transverse Section. 



278 THE BACTERIOLOGICAL WORLD. 

Claus, Text-book of Zoology, Protozoa, page 208, gives an 
illustration which is, I think, exactly similar to that I have 
described. He says, in describing the illustration : "Eainey's 
corpuscle from the flesh of a pig : ( a ) an animal inside a 
muscular fibre." He describes spores in a part more highly 
magnified. The appearance presented by sections of these 
muscle fibres are those that would be shown, when examined 
with a high power, if a fibre contained in its centre some sub- 
stance of a much higher refractive index than that of the 
muscle itself, and careful illumination leaves little doubt that 
these appearances are produced by the growth of mycelium in 
the fibre. 

After making a series of experiments with different 
reagents I have been able to show that portions of the rays are 
contained in the muscle fibres, and they are plainly made out 
with a high power as they stain quite differently to the myce- 
lium. — Pamphlet by the Author. 



THE MICRO-ORGANISM OF TRACHOMA.— SHONGOLOWICZ, 

[ TRANSLATED BY H. W. LOEB, M. D., ST. LOUIS, MO. J 

From the Centralblatt fur BaM. und Parasit. VIII -25. 

The writer proposed to attempt 'a discovery of a micro- 
organism in trachomatous tissue, and, in case one was con- 
stantly found, to determine exactly its nature. 

After a careful review of the previous literature upon 
this question, the writer reported his own observations. 

For experimentation he obtained the follicle contents of 
twenty-six patients with well-developed trachoma, in twelve of 
which it was necessary to cut the canthus. 

In the contents of the follicles Shongolowicz found only by 
a special method of staining ( a modification of the G-ram- 
Weigert procedure) bacilli, 1-2 jjl (.00004-00008 in.) long, 
and 0.3-0.5 /x (.000012-.00002 in.) in thickness, most of them 
straight, and a few bent ; they were seldom in pairs and were 



THE BACTERIOLOGICAL WORLD. 279 

not connected like chains. The ends of the bacilli were 
rounded off. These bacilli could often be demonstrated in the 
tissues in large quantities. 

For culture media he used 5 per cent, and 10 per cent, 
meat-peptone gelatine, 1 per cent, and 1% per cent, meat- 
peptone agar, coagulated ox-blood serum, potatoes and beef 
tea. The cultures were placed upon the follicle contents, 
which had been previously broken up. 

In this way the writer obtained three kinds of pure 
cultures : 

First. Micro-organisms seldom found in the conjunctival 
sac, and especially : 

( q, ) Three times, the potato bacillus. 

( b ) Twice, the hay bacillus. * 

( c ) Twice, the sarcina aurantiaca. 

Qd) Once, the sarcina alba. 

Second. Micro-organisms found somewhat frequently in 
the healthy as well as the diseased conjunctival sac, especially : 
(a) Twelve times, the staphylococcus pyogenes albus. 
( b ) Nine times, the staphylococcus pyogenes aureus. 
( c ) Three times, the staphylococcus pyogenes citreus. 
( d ) Three times, the staphylococcus cereus albus. 

Third. Micro-organisms, which in trachoma penetrate 
deeply into the conjunctival tissue ; these are found in the 
form of short bacilli in the seventh pure culture and correspond 
to the micro-organisms found by the writer in the degenerated 
trachomatous tissue. 

This last bacterium is developed very well, but slowly when 
the follicle contents are placed in blood serum, meat-peptone 
gelatine, meat-peptone agar, potatoes and beef tea. The bacilli 
in the form of hanging drops exhibit rapid motion, but they 
never appear as chains. Gentian violet is the best stain for 
the bacillus. Their length is 0.75-2.0 yu (.00003-. 00008 in.), 
and their breadth 0.3-0.5 ja( .000012-00002 in.). 

From the fact that the bacilli strongly interrupt the light 
at one or both poles, and that the color is intensified, the writer 
explains the appearance of micrococci as claimed in trachoma 
by earlier authors. 



280 THE BACTERIOLOGICAL WORLD. 

By inoculation of pure cultures upon the conjunctivae of 
cats and rabbits, a disease identical with trachoma sometimes 
appears. 

With respect to the constant presence of the bacilli in the 
altered conjunctiva in cases of trachoma, the writer 's opinion is 
that they are probably caused, not by micrococci, but by short 
bacilli. 



LEQTURE ON PHAGOCYTOSIS AND IMMUNITY. 



DELIVEKED AT THE TNSTITUT PASTEUR, DECEMBER 29, 1890. 
BY DR. ELIAS METSCHNIKOFF, 

Chef de Service de L'Institut Pasteur, Paris ; late Professor of Zoology 
in the University of Odessa. 



[Translated from the Original Manuscript by J. G. Adami, M. A. 

M. B. Cantab., John Lucas Walker, Student in Pathology 

in the University of Cambridge.] 

It is not possible to study the bacteriology of disease with- 
out noticing that, while in many cases the invading micro- 
organisms are to be found solely in the fluids of the body, in 
not a few affections they present themselves in the interior of 
certain cells, and this either partially — some being within the 
cells, others free in the blood plasma and the lymph that bathes 
the various tissues — or exclusively, all the bacteria that are 
visible being intracellular. Many of the facts bearing upon 
the terms of this relationship between tissue cell and micro- 
organism are now well known, yet it is worth while to recapitu- 
late the more important, in order to show that from them it is 
possible to gain a general law ; and what is more, that from a 
study of such facts some insight may be gained into the 
phenomena of immunity. 

It may, in the first place, be postulated that, whenever a 
micro-organism is discoverable within a cell, its passage thither 
has been by means of protoplasmic movements of amoeboid 
movements either on the part of the microbe, or of the cell 
itself. The first alternative is the rarer, although it certainly 
exists, and of this the malarial parasite affords an excellent 



THE BACTERIOLOGICAL WORLD. 281 

example ; for here in the amoeboid stage of its existence the 
hasmatozoon makes its way into the interior of a cell that 
possesses no active movements of its own, namely, the red 
blood corpuscle, and from the substance of this corpuscle the 
parasite gains its nourishment. Other sporozoa furnish 
instances almost equally good. More commonly, however, as 
in the case of all bacteria, where we have to deal with micro- 
organisms which, even when mobile, are destitute of protoplas- 
mic appendages, it is the cells which play the active part ; 
certain cells include the parasites. Of such, the amoebiform 
leucocyte of the blood and lymph is the most typical example, 
capable, as it is, of sending out pseudopodia in all directions, 
while a closely allied form is the cell of the splenic pulp. 
But there are also cells, as, for instance, those forming the 
endothelial lining of the vessels, which are very definitely 
fixed, which, nevertheless, can give off protoplasmic processes 
from their free surface, and so capture and include bacteria. 

All these may be spoken of as phagocytes, and may be 
divided into the two broad groups of fixed phagocytes ( endo- 
thelial cells, etc.) and free ( leucocytes ) ; not that the terms 
11 phagocyte" and "leucocyte" are synonymous, for of the 
latter three main forms may be distinguished, of which one is 
practically immobile and never takes up bacterid. This is the 
lymphocyte, characterized by its relatively small size, its large 
single nucleus, and the small amount of surrounding proto- 
plasm. The two remaining ( phagocytic ) forms are, first, the 
large uninuclear leucocyte, whose prominent nucleus is at 
times lobed or reniform, which stains well with aniline dyes 
and possesses much protoplasm and active amoeboid movements 
— the macrophage — and second, the macrophage, a small form, also 
staining well, but either multinuclear or with one nucleus in 
the process of breaking up. ( See Fig. 3.) If now we compare 
the endothelial cells with these, it is evident that their proper- 
ties connect them closely with the macrophage ; and, in fact, 
there is now little or no doubt that a very large proportion of 
the macrophages are of endothelial origin. 

Leaving aside the subject of amoeboid microbes and their 
life within animal cells, it is to the phagocytes and their rela- 
tion to the bacteria that I wish specially to draw your attention. 



282 



THE BACTERIOLOGICAL WORLD. 



Taking as wide a view as possible of this relationship, we 
can first determine that the more malignant the micro-organism 
the rarer is its presence within the phagocyte. Thus in those 
which of all diseases are the most rapidly fatal — in chicken 
cholera affecting birds and rabbits, in hog cholera (" cholera 
des pores''") given to pigeons and rabbits, in the anthrax of 
mice and other specially sensitive animals, in the "septicemie 
vibrionienne " * of guinea-pigs and birds, and in yet other dis- 
eases of peculiarly swift course — the corresponding bacteria are 
only very exceptionally to be found within the cells, but remain 
free in the neighborhood of their introduction, and thence 
invade the blood. For all the above-mentioned diseases are not 
localized, but, on the contrary, present the characters of general 
acute septicaemia, causing death within twenty to thirty-six 
hours, or, in certain cases, even within six hours. 




Fig. 1.— Staphylococcus pyogenes aureus contained in epithelial cell a, and leucocytes 
( microphages ) b b of alveoli of lung of rabbit ( Laehr ), c c vacuoles. 

And when we pass to those diseases in which the bacteria 
are to be found either in part or almost wholly within the 
phagocytes, the same law still applies ; for in such cases the 
disease has lost its suddenness, tending to have a slower course,, 
or, indeed, to be of a chronic nature. Even in those affections 
in which an acute course is accompanied by considerable phago- 
cytosis, the fatal termination is far from occurring at the same 
early period as in the diseases recorded above. Thus mouse 
septicaemia, characterized as it is by frequent intracellular 



* The disease produced by a micro-organism, which in appearance and methods 
of growth is curiously like Koch's cholera spirillum. Gamaleia, its discoverer, 
named it " Vibrio Metschinikovi " ; and as such, in place of M. Metschnikoff's own 
vaguer " septicemic vibrio, " it will be spoken of in later reference to its properties. 
The disease it originates will be spoken of as vibrionic septicaemia. "—Trans. 



THE BACTERIOLOGICAL WORLD. 



283 



bacteria, has a duration in the mouse two and a half times as 
long as that of anthrax in the same animal. But in general a 
well-marked phagocytosis is associated with diseases presenting 
an essentially chronic development ; it is in affections such as 
tuberculosis, leprosy, rhinoscleroma, glanders, that the specific 
bacteria are most readily taken up by the phagocytes ; it is here 
that at the seat of the disease we meet with innumerable macro- 
phages — epithelioid cells in which lie the individual micro- 
organisms. 

Further if we consider the phenomena associated with the 
resolution of an infectious disease, this inverse relationship 
between the malignancy of the malady and the occurrence of 
phagocytosis is, if possible, yet more clearly demonstrated. 
Notice, for instance, what obtains during the progress of relaps- 
ing fever, a malady still fairly common in Eussia and other 
Sclavonic countries, and one which, while presenting many 
difficulties to the bacteriologist in that the specific spirochaete 
has so far resisted cultivation, and in that it cannot be commu- 
nicated to the ordinary animals of the laboratory, is neverthe- 
less in many respects not ill adapted for our present purpose. 
Here, during the sudden access of the fever, the spirilla are 
present in the blood in enormous numbers ; they all are free in 
the plasma, and not a single intracellular spirillum is to be met 
with. During the apyretic ( and in the monkey this is, at the 




of 



~-:CT 



Fig. 2.— Resolution of acute infectious disease (relapsing f ever ), spleen pulp of 
monkey ( Macacus erythr. ) , showing ( a ) a microphage, multinuclear, with 
incepted spirochaetes ; ( b ) solitary, and ( c ) forming dense tangle, {d d) 
nuclei of splenic tissue ( Zeiss. 1-18 ocular 4 ; X 1515 dim. ) . 



284 



THE BACTERIOLOGICAL WORLD. 



same time, the stage of resolution) not a single free spirillum is 
discoverable in the blood, while the phagocytes of the spleen 
contain the microbes. The like phenomena repeat themselves 
in all those cases where it is possible to follow the fate of the 
micro-organisms of acute disease during the stage of recovery. 
Thus rats and pigeons very frequently survive an attack of 
anthrax, and, where this occurs, the bacteria, which at the 
commencement of the disease were for the most part free, now, 
during resolution, are for the most part included within 
leucocytes and splenic phagocytes. 

Nor is this all ; analogous phenomena as a rule attend 
immunity, which most often is but recovery in operation from 
the very onset of a disease. The more closely one studies this 
condition of immunity, the more is one led to the conviction 
that immunity and recovery are very intimately connected ; that 
one can pass by slight gradations from the resolution of disease 
to the production of immunity. So it is that, in inoculating 
refractory animals with the microbe to whose action they have 
been rendered immune, it is found that the parasite begins to 
develop, but that from the onset a reaction on the part of the 
organism shows itself, accompanied by a considerable emigration 
of leucocytes which soon include the bacteria in great numbers. 




..■c; 



Fig. 3.— Anthrax of pigeon (an animal but slightly susceptible to the disease), to 
show stages of destruction of bacilli by phagocytes. 1 and 2, macrophages ; 
1, from exudation from eye of refractory bird ; 2, from muscle of region of 
inoculation of bird that succumbed ; 3, 4, 5, microphages— all from eye 
twenty-seven hours after inoculation; a, a, unaltered bacilli ; 61, b2, bd, bacilli 
becoming more and more degenerated and indistinct; c c, debris of bacilli 
(Zeiss 1-18, ocular 3). 



THE BACTERIOLOGICAL WORLD. 285 

This relationship of phagocytosis to acquired immunity is 
in the highest degree instructive. Where a given species of 
animal is specially sensitive to the onslaught of one or other 
micro-organism, there, during the course of the disease, the 
phagocytes are inoperative, including none, or almost none, of 
the bacteria. On the other hand, when by previous vaccination 
these animals have been rendered refractory, their phagocytes 
have acquired the property of including the same bacteria. As 
an example of this, I may cite the action of the bacillus of 
anthrax and of the vibrio Metschnikovi. In ordinary rabbits 
the development of anthrax is only followed by a very feeble 
phagocytosis, while in vaccinated rabbits this phagocytosis is 
very extensive. Corresponding, but yet more strongly marked, 
differences are to be made out between the unvaccinated 
guinea-pig — an animal most readily affected by the vibrionic 
septicemia — and the guinea-pig vaccinated against the same ; 
after inoculation with the vibrio Metschnikovi, none of the 
vibrios are to be found in the cells of the former ; in the latter 
the phagocytes are simply replete with the microbes. 




1 

Fig. 4. --Phagocytes, macrophage and microphage, to show stages of digestion and 
destruction of bacilli, from spleen and eye respectively of white rat with 
anthrax. In 1, part of the bacillus is unaffected, but a vacuole has formed 
around the other part, which further has now lost the power of taking the 
stain. In 2, various stages are seen, the bacilli passing through the granular 
badly staining, to the vacuolated unstained, until finally but faint "shadows" 
are observable (Zeiss 1-18, oc. 3). 

The facts enumerated thus far would seem to prove that 
there exists a certain antagonism between the microbes and 
the phagocytes, and this view is confirmed by the fact that 
in general the microbes find the interior of the phagocytes 
an unfavorable medium for their development and con- 
tinued existence. Very often it is possible to determine abso- 
lutely that the parasites are killed within the phagocytes • 



286 



THE BACTERIOLOGICAL WORLD. 



after inoculating refractory animals with bacteria, an afflux of 
white corpuscles toward the region of inoculation, followed by 
the inclusion of the bacteria and by their death, is seen to 
occur. These stages can be well followed where the anthrax 
bacilli are taken into the phagocytes of animals that are, or 
have been rendered, immune. They occur also with a long 
series of other micro-organisms studied in this connection, and 
among others in the case of the tubercle bacillus invading 





Fig. 5.— Two giant cells seen under high magnification ( X 1515 diam.) from a rodent 
the spermophile, inoculated with tuberculosis, to show stages in the destruc- 
tion of the bacilli, a, unaltered bacillus; b, bacillus staining badly, and with 
greatly thickened capsule ; c, bacillus granular and breaking up; d, e, 
" shadows." 



THE BACTERIOLOGICAL WORLD. 287 

animals that are more or less resistant. The giant cells of 
tuberculosis are, in fact, huge multinuclear phagocytes ; and 
here the intracellular destruction of the bacilli is the more 
clearly demonstrable, inasmuch as the micro-organisms exhibit 
such very evident signs of degeneration ; the bacilli swell, their 
enveloping membrane becomes much thickened and highly 
refractive, and, in time, the contents lose their power of fixing 
the staining material, so that, eventually, nothing is left but 
slightly yellowish forms, recalling, in proportions and positions, 
the enlarged bacilli, and these shadowy bodies unite into small 
masses of an amberlike appearance. Analogous transforma- 
tions never being observable outside the phagocytes— that is, 
to say, either in cultures or in caseating masses — these changes 
may well be regarded as due to a specific action upon the part 
of the giant cells. 

The broad fact that the invasion of the organism by 
microbes most often induces, on the one hand, an inflamma- 
tory reaction with its associated emigration of leucocytes, and* 
that, on the other hand, the phagocytes are capable of includ- 
ing and destroying the invaders, leads us to admit that the 
afflux of phagocytes to the invaded region, and their bactericidal 
properties, are mechanisms which serve to ward off bacterial 
attack and to maintain the integrity of the organism. Where 
the phagocytes do not, either immediately or eventually, inter- 
vene, but leave the field free to the microbes, these last 
multiply without hindrance, and succeed in killing the animal, 
within, it may be, an excessively short period. Thus the micro- 
organism of hog cholera, which is left quite untouched, kills 
the pigeon in the course of a few hours — often, within five 
hours after inoculation ; chicken cholera kills not only pigeons, 
but also rabbits, in an equally short period. In other diseases 
in which the phagocytes appear upon the scene in relatively 
large numbers, and even include the micro-organisms, the latter 
gain the day whenever and wherever the phagocytes are incapa- 
ble of destroying them or of preventing their growth. 

This manifest bactericidal action is to be compared with 
the phenomena of intracellular digestion characteristic of 
amoeboid cells in general, and of leucocytes and other microbic 



288 THE BACTERIOLOGICAL WORLD. 

phagocytes in particular. These cells have the power of digest- 
ing, with ease, red corpuscles and other organized elements ; 
just as have the amcebse proper, and other protozoa. Among 
these last are many which have been found to include and 
transform bacteria in exactly the same way as do the phago- 
cytes of the higher animal. — The British Medical Journal. 

( To he continued,) 



SCIENTIFIC GLEANINGS. 



ON THE -FORMATION OF STARCH INTO DEXTRINE BY THE 

BUTYRIC FERMENT. 



BY M. A. VILLIERS. 

(Note to the Academy of Sciences, Paris.*) 

Having undertaken the study of the action of the figured 
ferments on the hydrates of carbon, in divers conditions, I will 
give here the first results relative to the action of the butyric 
ferment ( bacillus amylobacter ) on potato starch. 

It is easy to transform the amylaceous matter into dex- 
trine under the action of this, ferment. 

In large flasks, we introduce the starch of a potato and 
water not distilled, in the proportion of 50 gr. to 1 litre, the 
starch having been previously diluted with care. This mass is 
afterwards transformed into starching material by a jet of water 
vapor directed into the bottom of the flasks, agitating con- 
stantly, until the temperature rises to 212° F. After the con- 
densation of the vapor, the flasks must be about refilled, in a 
manner that it only leaves a little volume of air on the surface 
of the liquid. 

Then, the starch formed is inoculated with a few cubic 
centimetres of a culture of bacillus amylobacter. The flasks 
are then stopped with a stopper of sterilized cotton and depos- 
ited in an oven at 104° F. to remain some days. In these 



* Comte's Rendus y page 435, February 23, 1891. 



THE BACTERIOLOGICAL WORLD. 289 

conditions, the spores of the butyric ferment, which, as has 
been shown by Mr. Van Tiegham, resist easily a temperature of 
212° F. develop themselves rapidly. In inoculating the starch 
at 212° F. with the starch microbe, the development of foreign 
germs is more easily prevented. * * * 

At the end of twenty- four hours, the starch is, in general, 
liquefied. Fermentation is allowed to continue until the liquid 
gives no more blue or violet reaction by iodized water. This 
result is attained more or less rapidly, after two to four days, 
sometimes only at the end of many days, owing to the existence 
of lumps difficult to attack. It is necessary, therefore, to make 
a starchy material as homogeneous as possible. 

The bacillus presents itself, at the beginning, under the 
form of straight, short rods, very mobile. At the end of 
the transformation, it has uniformly thickened extremities, in 
the characteristic form of tadpole ; the bacilli are then com- 
pletely immobile. From this moment the products of the 
fermentation are modified no more. 

Little, gaseous bubbles disengage themselves during this 
transformation of the starch, but the quantity of the gas 
disengaged is so small that it is impossible to gather any of it. 

The liquid thus obtained is very slightly acid, and 
presents clearly the odor of butyric acid, but contains only 
an insignificant quantity of this latter (about 0.3 parts to 100 
of starch ) . 

Besides certain substances, which are also found in very 
small quantities, and to which I will return shortly, the prin- 
cipal products of this fermentation are constituted by dextrine, 
resisting the attacks of the bacillus amylobacter, at least, in the 
presence of other products, which are simultaneously formed. 

We obtain them in precipitating the filtered and evap- 
orated liquids and purifying them by further treatment with 
alcohol. The dextrine thus precipitated, added to that 
remaining in solution in the alcohol, represents the larger part 
of the starch employed. 

Dried, they present themselves in the form of a perfectly 
white mass, light, friable, very greedy for water, which 
combines therewith, producing heat ; it has a little sweet flavor, 
and is constituted by a mixture of different dextrines, as is 



290 THE BACTERIOLOGICAL WOELD. 

shown by the variation of the rotary powers corresponding to 
the products of different operations, or even of fractions of 
precipitates of the dextrine resulting from a single operation. 
These powers varied from + 312.8° F. to + 405.5° F. 

They transform themselves with difficulty into glucose by 
the action of water and acids. The transformation under the 
action of water does not seem possible at a cold temperature ; 
it is very slow at 212° F. and hardly commenced at the end of 
forty-eight hours. Under the action of dilute sulphuric acid, 
it requires about a day at 212° F. 

The iodine colors in red, the dextrine with the highest 
rotary power ; the intensity of the coloration diminishes in 
them in proportion with the decrease of the rotary power : 
those in which this power is the lowest are not colored by the 
iodine. 

They reduce cupropotassic liquor, and their reducing 
power is the greater that their rotary power is the more feeble, 
as we can judge by the following results, representing the 
weight of glucose which would reduce the same volume of 
reagent as 100 parts of dextrine : 

Kotary power. Eeducing power. 

156 28.9 

175.4 11.3 

207.5 5.0 

New researches are necessary to decide if the dextrines 
are identical, or not, to those obtained by the action of the 
acid, or under the influence of diastase. But, at all events, 
their production, in the complete absence of maltose and 
glucose, is worthy of attention in the study of the constitution 
of amylaceous matter. On the other hand, this absence of 
glucose and maltose seems to show that the butyric ferment 
determines the transformation of starch into dextrine directly, 
and not by the intermediary of a diastase secreted by the 
organized ferment, diastase which should, judging from the 
analogies known, determine the saccharification of a larger or 
smaller quantity of dextrine. 



THE BACTERIOLOGICAL WORLD. 291 

THE AMMONIAC AL FERMENTATION OF URINE. 



BY VERANUS A. MOORE, M. D., 

Washington, D. C. 

The fact is well known that urine undergoes putrefactive 
changes when it is allowed to stand for a considerable length of 
time at the ordinary temperature. In following these in the 
order of their occurrence we find that the clear urine first 
becomes faintly, then heavily, clouded, and finally, after several 
weeks standing, it clears with the formation of a considerable 
quantity of a grayish, more or less granular, sediment. The 
color of the cleared urine is always several shades darker than 
that of the fresh liquid. A further examination shows that its 
acid reaction is converted into an alkaline one simultaneously 
with the occurrence of the clouded appearance, and with this 
change in its chemical reaction the odor becomes penetrating 
and ammoniacal in character. The source of the ammonia 
was explained by the discovery of urea, and later investigations 
made by Prout showed that the urea was converted into car- 
bonate of ammonia by a process of hydration. By i( ammoniacal 
fermentation of urine," therefore, is meant that change in 
urine by which its urea is transformed into carbonate of 
ammonia. * * * 

BIOLOGICAL HISTORY. 

In 1860, Pasteur showed that in decomposing urine the 
transformation of the urea into carbonate of ammonia was due 
to a micrococcus which he found and designated as Torula 
ammoniacale. ( *Van Tiegham, Miquel, Leube and Flugge 
described several forms of urine ferments.) * * * 

To summarize, we have as organized ferments of urine 
already described six micrococci (Pasteur's, Flugge's and 
MiquePs), seven bacilli (eight if Miquel 's bacillus urea (5 and 
Leube 's bacterium urea are not identical) and one sarcina 
that are more or less active in the transformation of fresh into 
ammoniacal urine. 



* In parenthesis by the editor. 



292 THE BACTERIOLOGICAL WORLD. 

In considering the source of the organisms, we find that 
Pasteur, Van Tiegham and Leube isolated- their germs from 
decomposed urine. Flugge does not give the material from which 
he obtained his mic. urea liquefacians. Miquel found the germs 
he described in the atmosphere, soil and water. Pasteur also 
found his torala ammoniacale among the germs isolated from 
the atmosphere. 

In view of these facts, I have confined my investigations 
thus far principally to those forms of bacteria that are to be 
found within the healthy urethral canal. The object of such a 
limit was to determine, if possible, whether or not there are 
certain bacteria that are constantly present in this canal which 
act as exciting agents in the transformation of fresh into 
ammoniacal urine. If this question can be definitely settled it 
may be effectual in explaining the causation of certain cases of 
cystitis which are rapidly developed after the introduction of 
bougies and cathethers. Dr. Eicard finds that if these 
instruments are made aseptic that their introduction is harm- 
less, providing the urethral canal is not infected ; but, if the 
urethral canal is already infected, it must previously be washed 
with a saturated solution of boric acid in order to avoid a subse- 
quent more or less acute cystitis. It seems reasonable to sup- 
pose that if germs are found constantly within the healthy 
urethra that are active in producing ammoniacal urine that this 
canal as well as the instruments should always be thoroughly 
disinfected before any operation is attempted. 

METHODS EMPLOYED IN COLLECTING AND EXAMINING URINE. 

The methods employed, both in collecting the fresh urine 
and in its subsequent examination, are of much importance and 
worthy of careful consideration. The following precautions 
were rigorously adhered to in collecting the fresh liquid : The 
glands and external urinary meatus were carefully disinfected 
with either a 5-per-cent. solution of carbolic acid or 1-1000 
solution of corrosive sublimate. The urine was passed directly 
into a sterilized Erlenmeyer flask, the mouth of which was pre- 
viously flamed and the cotton-wool stopper replaced immedi- 
ately after urination. Care was also taken to avoid dust and 
currents of air. A few cubic centimetres were immediately 



THE BACTERIOLOGICAL WORLD. 293 

removed from the flask by means of a sterile pipette for a pre- 
liminary chemical examination. Only normal acid urine was 
retained for the bacteriological examination. 

For the isolation of the germs the well-known Koch 
gelatine-plate method was employed. Es march rolls were at 
first tried, but on account of the presence of liquefying germs 
they were abandoned. In several cases a plate culture was made 
from the fresh urine by adding from K to 1 cc. of it to a tube 
of liquid gelatine by means of a sterilized pipette, and, after 
thoroughly mixing, pouring the gelatine upon sterile glass plates. 
These were kept at a temperature of 65° to 75° F. These plates 
developed from one to four colonies, occasionally more, often 
none at all, showing how comparatively few bacteria there are 
in freshly voided urine. The flasks were allowed to stand at 
the ordinary temperature (65° to 75° F.) for from twenty-four 
to forty-eight hours, when the urine became clouded and either 
neutral or alkaline in reaction. A microscopical examination 
at this time showed a greater or less number of bacteria present. 
Plate cultures in gelatine were then made. One small platinum 
wire loop (holding from -fa to -£ cc.) of the fermenting urine 
was added to a tube containing about 10 cc. of liquid gelatine. 
After thoroughly mixing, from two to four loops of the first 
gelatine were added to a second tube of gelatine. The gelatine 
was poured on sterile plates. (These were carefully protected 
from external contamination by means of flamed bell jars. 
Sterile "double-dishes" were also used). They were kept at a 
temperature varying from 65° to 75° F. The number of loops 
taken for the first tube of gelatine depended upon the number 
of germs seen in the microscopical examination. When the 
germs were exceedingly numerous the first dilution was made 
in a tube containing about 7 cc. of sterile distilled water. The 
first plate usually developed innumerable colonies after twenty- 
four to forty-eight hours. The second plate as a rule developed 
from fifty to three hundred colonies. As soon as they were 
sufficiently developed they were examined both macro scopically 
and microscopically and a subculture made in sterilized urine 
and gelatine from a colony of each apparently different form. 
The plate cultures were repeated after the urine had reached its 
maximum degree of alkalinity, and again after it had been 



294 THE BACTERIOLOGICAL WORLD. 

standing for from two to three weeks. In a few cases parallel 
plates in agar were made, but no germ was found to develop in 
this medium that did not grow on the gelatine. The subcultures 
were carefully examined, and all those in which the urine 
culture became alkaline were retained for more careful study. 
By this process I have isolated from urine about twelve forms 
of bacteria, mostly micrococci, of which five, on account of 
their property of producing ammoniacal fermentation in urine, 
will receive a somewhat detailed description. Of the remaining 
forms, one was a streptococcus which was present in every 
specimen of urine examined, two were bacilli, and the 
remainder were micrococci. The bacilli were very rarely found. 

CULTURE MEDIA EMPLOYED IN" TESTING THE REACTION" OF 

THESE GERMS. 

For a preliminary examination of these germs the ordinary 
culture media were employed, but, in order to test their power 
to transform urea into carbonate of ammonia, media containing 
that substance were necessarily resorted to. Of those sterilized 
normal urine seemed to be the most practical, as well as to 
afford a sufficient test for the property of the germs in question. 
Other media, such as acid-beef-infusion-peptone and gelatine 
plus urea, were also employed. Of the ordinary media, nutri- 
tive gelatine was the most important from a differential 
standpoint. 

i. Sterile urine. — This was prepared by distributing 
fresh acid urine in sterile culture tubes, about 10 cc. in each 
tube. As the urea is broken up by a high temperature the 
tubes were placed in a large water-bath and heated to a temper- 
ature of 149° F. for two hours each day for four consecutive 
days. Their sterility was tested by allowing them to stand for 
several days in an incubator at a temperature of 96.8° F. 
The urine remained in every instance clear and acid in reac- 
tion. 

2. Acid-bouillon plus urea. — This was prepared by 
macerating 100 grams of finely-chopped beef ( freed from fat ) 
in 200 cc. of distilled water. After standing for from eighteen 
to twenty-four hours in a cool place it was strained through a 
coarse linen, and to the simple beef-infusion ^ per cent. 



THE BACTERIOLOGICAL WORLD. 295 

sodium chloride and 1 per cent, peptone were added. It was 
then boiled for thirty minutes, cooled, filtered, and % per cent. 
urea added. The liquid was then distributed in sterile culture 
tubes and sterilized by heating at 149° F. for three hours each 
day for four consecutive days. It remained clear and acid after 
standing for several days at a temperature of 96.8° F. 

3. Acid gelatine plus urea. — This is prepared pre- 
cisely in the same manner as the acid bouillon, with the 
addition of 10 per cent, gelatine, which is added before the 
beef-infusion is boiled. The urea is added to the liquid gelatine 
after it is filtered. It is preserved in sterile, cotton-plugged test 
tubes, and sterilized by the same method as the bouillon, after 
which it remained clear and acid in reaction. 

[To be continued.] 



296 



THE BACTERIOLOGICAL WORLD. 



M M ■-■*£&■* 








Ifi-ft 



Fig. 1. 



<*:" 



i I 



£¥' i 



fi .. . ,|- 7. i».< ^IP^S^V 







Fig. 2. 



Fig. 3. 





Fig. 4. 




fcl -w Ma I 




Fig. 5. 



THE BACTERIOLOGICAL WORLD. 297 



DESCRIPTION OF PLATES. 

Plate I. 
Gelatine needle cultures of the micrococci described. 
Fig. 1. Micrococcus a — Culture seven days old. 

"3. " c " six weeks old. 

"4. " 

" 5. 



D 


a a 


E 


(( a 




Plate II. 



Drawings made from cover-glass preparations of urine cultures, one 
day old, of the micrococci described. The preparations were dried, 
passed three times through a flame, and stained for a short time with 
alkaline-methylene-blue. The position of the germs in the field was 
determined by means of Abbe's camera lucida, Zeiss 2 mm. apochro- 
matic lens, No. 4 eye-piece. 

Fig. 1. Micrococcus a. 

" 2. " B. 

" 3. " c. 

" 4. " D. 

" 5. " E. 



298 



THE BACTERIOLOGICAL WORLD. 




Fig. 4 Fig. 5 

Proceedings of the American Society of Microscopists. 



THE BACTERIOLOGICAL WORLD. 299 

TECHNIQ TIE. 



RAPID STAINING OF TUBERCLE BACILLI FOR DIAGNOSIS. 

BY PROF. V. A. LATHAM, ANN ARBOR, MICH. 

The methods I prefer are Gibbes' Double and his Magenta, 
and, as a counter stain, Ziehl Neelsen's; though for permanent 
preparations I believe the two former methods the best, for by 
Ziehrs the bacilli lose the rose tint and become black. 

To double stain at once : — 
To make the solution : 

Jfy Rosaniline Hydroc > 3 grams 

Methyl Blue = . 2 grams 

Rub up fine in a glass mortar, then dissolve aniline oil 6cc. 
in rectified spirits or alcohol 20cc; add very slowly to the stains, 
rubbing up well till dissolved ; then slowly add distilled water 
20cc, and keep in a stoppered bottle and date it. To 
stain very quickly, pour in a thin watchglass a little of the 
stain ; place the cover on which the caseous nodules of 
sputum have been spread evenly and thinly, * and then dried, 
sputum side down and press under with a needle. Then heat 
till it steams and fluorescence is seen, then let it stand covered 
for five minutes, longer if possible. When stained, wash well 
in plenty of alcohol till quite clean and all red color off ; let dry 
and on the cover place a drop of C. Balsam, and lay on a slide. 
If the best results are desired the stain should be used cold and 
left for an hour. This method does away with all the acids 
and keeps well, and is certain if bacilli are present. It can be 
used for micro-organisms and bacilli in tissue, and is a gain as 
the tissue does not shrivel, — Lancet, May 5, 1883. 

magenta stain" {Dr. Gibles). 

1^ Magenta 2 grams 

Aniline oil 3 cc. 

Alcohol (.830) 20 cc. 

Aqua, dist .- 20 cc. 

Dissolve the oil in the alcohol and shake. Rup up crystals 
very fine with a little alcohol until all dissolved, then add 

-And, by aid of a sputum spreader, got from Eberbach's, Ann 
Arbor, Mich., this is easy. 



300 THE BACTERIOLOGICAL WORLD. 

distilled H 3 while stirring; keep stoppered, dated and 
labeled. 

To use slain. — Spread sputum as above, allow to dry in 
air protected from dust, or by heat if in a hurry. In any case, 
fix to cover by heat. Pour out magenta in a watchglass, and let 
the sputum cover lie face down for 20 to 30 minutes. While 
staining, make or keep on hand a 25-per-cent, to'33-per-cent. 
solution of HN0 3 (0. P.) in distilled water. Eemove cover 
from stain and thoroughly decolorize in acid, then wash well 
in water distilled, and if the red color returns wash repeatedly 
in the acid and again in distilled water. Then stain in a 5-per- 
cent, solution of methyl blue, green or iodine green in distilled 
H 2 0, and leave for 5 to 10 minutes ; wash in distilled water 
until no color comes away; drain off water; wash in alcohol; let 
dry and mount. If it is desired to study the structure of bacilli, 
use a watery solution of chrysoidin in place of blue, etc. Care 
must be taken that the sputum is the first coughed up in 
the morning, as later it is often only from the back of the 
throat. Heating the stain will facilitate the process. The 
Germans, I believe, are giving up the value of bacilli as a reli- 
able diagnostic point. 

For actinomycosis. — A solution of Spiller's blue is recom- 
mended, and mount in C. Balsam, the fungus being unstained ; 
or Gibbes' double-stain, if left in it long enough. 



THE BACTERIOLOGICAL WORLD. 301 

REMEDIES AND CLINICAL NOTES. 



CAMPHO-PHENIQUE AND LISTERINE IN ALVEOLAR AB- 
SCESS. 

I was called hastily by a neighboring dentist to see a patient, Miss 
M., aged 24, who, he stated, was having a severe hemorrhage from the 
gum where a tooth had just been extracted. I found a good-sized 
abscess under the lower maxilla which, she stated, had been there for 
about three months. From three small incisions — to avoid scars — I 
gently pressed out blood and pus, and applied: 

J$_ Campho-phenique 5 n J« 

Lanolin . 

Vaseline aa. q. s., ad § j. 

M. ft. ungt. Sig. : Apply night and morning, previously bathing 
the affected part with warm water. Also ordered internally: 

I£ Quin. sulph gr. xlviij. 

Tinct. ferri mur § ss. 

Glycerine § j . 

Aquae, q. s. , ad § iij. 

M. ft. Sig. : Two teaspoonfuls in water three times a day. 
Also ordered: 

]£ Fl. ext. hydrastis can 5 ^ v * 

Li&terine 5 j. 

Glycerine g j. 

Aquae, q. s. , ad § iv. 

M. Sig.; Use as a mouth wash every two or three hours. 

Patient returned to my office in about ten days with abscess almost 
cured, and looking much relieved in body and mind. I am continuing 
the same treatment. I omitted to say that I am relieving a congested 
portal circulation. 

T. B. Kramer, M. D. 

Washington, D. C. — Medical Sun. 



A DETERGENT AND ANTISEPTIC WASH IN SCARLATINA 
AND DIPTHERITIC SORE THROATS. 

I£ Acid. Boracic ^ss. 

Chloral Hydrat gr. x. 

Glycerine ^ iv. 

Katharmon * ijss. 

Mi Sig. : Dilute with water and thoroughly cleanse the fauces 
and ulcerated surfaces every 2 hours. 

Dr. R. M. King, 
( Prof. Beaumont Med. College, St. Louis, Mo. ) 



302 THE BACTERIOLOGICAL WOELD. 

FOR NASAL CATARRH. 

J$_ Resorcine . g r. x. 

Glycerine , § i. 

Katharruon q. s. ad § iv. 

M. Sig. : For spray. 

Wm. Porter, M. D. 

St. Louis. 



MAMMIT1S. 



!£. Tarro- Petrolene q. s. 

Sig. : Apply in gentle friction over the inflamed organ twice or 
three times daily after washing and wiping dry. Leave a good coat 
of the ointment on the surface. 

N. B. This treatment is very successful. 

A. Rouif, V. S. 
St. Louis, Mo. 



QUERIES AND ANSWERS* 

Under the above heading we will answer, as briefly as possible, 
short, concise rational questions of subscribers, concerning bacteriolog- 
ical subjects only, as far as accommodating collaborators and the edi- 
torial management may be able to. 

Q. Who is the father of the theory of phagocytosis, Pasteur or 
Metschnikoff ? Med. 

A. The last-named scientist is the man. He is a Russian, for- 
merly of Odessa, now Chief of Pasteur's Institute, Paris. 

Q. To what kind of cells is the property of digesting bacteria, 
etc.,— phagocytosis ascribed? Med. 

A. To fixed cells as those of the broncliial epithelium, for instance, 
and to cells of the order of leucocytes. 

Q. How are specimens preserved ( not mounted) in glycerine, and 
Ireated afterwards if desired to cut and stain ? J.N. 

A. Glycerine, by its avidity for water, distorts specimens. How- 
ever, certain kinds may be preserved and hardened in it, and then 
sections made directly without further treatment. Before staining 
the sections should be a long time in successive clean-water baths, 
treated with alcohol, and washed again until all traces of glycerine 
have disappeared. Specimens may be preserved in glycerine in using 
first 2 parts of it to 1 part of water, transferring after a week or so to 
purer and then later to perfectly pure glycerine. Three parts glycerine, 

*The 7th, 8th, 9th and 10th questions are kindly answered by Prof. T. J. Burrill. 



THE BACTEEIOLOGICAL WORLD. 303 

2 of water and 1 of alcohol as first fluid is also used, followed with 
fluids of purer and purer glycerine. At best, we have found glycerine 
an inferior preservative for subsequent convenient manipulation of 
tissues. 

Q. Is Bismark brown the same as Vesuvin? H. L. P. 

A. Both are derivatives of diazobenzine and are consequently 
closely related if not identical, for an author states that the first named 
is partly soluble in distilled alcohol, and the latter is soluble in water. 
However, we found them about equally soluble in water, etc., and 
believe them the same reagent under a different name. 

Q. Why is it that experimenters in aniline do not give the name, 
number and maker so that the dye can be obtained at some future 
period ? R. James. 

A. Experimentalists, please answer. It would help the work 
along materially if the suggestions of this question were put in prac- 
tice. 

Q. Can anyone advise me regarding a camera, for both landscape 
and micro-photography, which is light and suitable for a lady's use for 
the former work ? B. Taylor. 

A. Will some one answer ? We know of no combination of the 
kind. 

Q. What is the best way to clear specimens of celloidin imbedding 
on a slide after cutting to get the tissues clearly stained ? 

A, It is usually best not to remove the celloidin from sections 
imbedded in this substance. If trouble is found it is probably from 
want of proper clearing, i. e., making transparent. Try the following: 
Cut sections, using 80 per cent, alcohol during the operation. Place 
short time in 95 per cent, alcohol, then remove to slide ; stain "with 
alcoholic solution ; clear with origanum oil, three parts, oil of cloves, 
one part ; mount in balsam. 

If, however, it is desired to remove the celloidin mass, use oil of 
cloves pure. If it proves necessary to fix sections on slide, use white 
of egg 50 cc, glycerine 50 cc. and salicylate of soda, 1 gram, well 
shaken together and filtered. Spread thinly on slide with brush, and 
lay sections on at once. Ninety-five per cent alcohol will now solidify 
the albumen and wash out the glycerine, which latter must be effect- 
ually disposed of if the object is to be mounted in balsam. 

A still easier method of fixing such sections to slide is available 
when the celloidin is not to be dissolved out, viz. : Simply by blowing 
upon them some ether vapor from a bottle, arranged like a commo n 
wash-bottle, containing sulphuric ether. This partially dissolves the 
celloidin and causes the sections to adhere. 

Q. How is the condenser used to get the exact focus so that an 
amateur can understand it ? 

A. Center the condenser by the use of a two-inch objective, or 
by removing both eye-piece and objective and looking down the tube. 



304 THE BACTERIOLOGICAL WORLD. 

I screw in an iris diaphragm in place of the objective, and remove 
both lenses from an eye-piece having a cap ; then by opening and clos- 
ing the diaphragm while looking down the tube, exact centering can 
be accomplished. Some condensers are furnished with a metal cap 
with central pin-hole — all ought to be so provided, for this matter of 
exact centering is very important. Now, tu adjust focus of condenser, 
use a lamp set at least two feet at one side, edge of flame towards 
plane mirror. See that the latter is central, and reflect the light up 
through condenser. Lay on the stage a slide having a thin paper 
label, with the latter over the center of the condenser. A bright spot 
of light will show on the paper, and the condenser is properly focused 
when this is as small as it can be made — almost a mere point. If it is 
daytime, and light from a large window is used, the best focus is 
shown by the image of the window-bars, etc., on the paper label. Of 
course anything acting as a screen will serve the purpose of the 
paper label, but it ought to be on the upper surface of a slide of 
medium thickness. 

Q. What is the best way to correct and adjust collar objectives ? 

A. Select a small dense particle of something under the cover for 
which the correction is desired. Most mounted slides will be found to 
have some little body which will answer for this purpose ; but, to 
experiment, rub up a little charcoal or take the spores from the sooty 
substance on corn called smut. Mount a very little, such as these, in 
water and examine under the desired objective. Now move fine 
adjustment up and down from the proper focus. A dark ring will be 
observed around the object swelling in diameter as the objective is 
moved. When the collar is properly adjusted the size of this ring 
( coma ) will be equal on both sides of the focus. The shade is usually 
deeper on one side than the other at best, but the enlargement ought 
to be the same. 

Q. Why is it aniline varies so in its action, especially in staining 
T. bacillus when mixed in staining fluids ? 

A. In my experience it is not the aniline oil ( provided this is 
freshly mixed ) which varies in the staining of bacillus tuberculosis so 
much as it is other things in the manipulation. In Koch's original 
method he used one part of potash to ten thousand parts of water in 
connection with methyl-blue. This minute proportion of alkaline sub- 
stance should show the necessity of having the staining fluid just 
right. The least acidity spoils the result. One of the sources of 
trouble is in the water used. My results have sometimes been better 
with rain water than with the distilled water obtained at a chemical 
laboratory, but subject to absorption of various gases. 

Another cause of failure is in the overheating of the sputum in dry- 
ing. Too long immersion in very hot stain also seems detrimental. 



THE BACTERIOLOGICAL WORLD. 305 

All this is upon the assumption that the irregularity observed is 
with the same sample of aniline oil. There is much variation in the 
commercial substance sold under this name. 

Q. What is the method and object of mounting micro-organisms 
in tissues in monobromide of naphthaline ? 

A. See Journal of the Royal Microscopical Society, 1880, p. 1043 
(Abbe, and others) for accurate reply. Will gladly publish some one 
else's opinion on the point. 



LABORATORY NOTES. 



FIRST ANNUAL REPORT NEW YORK PASTEUR INSTITUTE. 

Dr. Paul Gibier, Director of the New York Pasteur Institute, begs 
to inform you of the results of the preventive inoculations against 
hydrophobia performed at this Institute during the first year of its 
existence (February 18, 1890, to February 18, 1891), — 828 persons, 
having been bitten by dogs or cats, came to be treated. These patients 
may be divided in two categories: 

First. For 643 of these persons it was demonstrated that the 
animals which attacked them were not mad. Consequently, the 
patients were sent back after having had their wounds attended 
during the proper length of time, when it was necessary. 

Second. In 185 cases the anti-hydrophobic treatment was applied, 
hydrophobia of the animals which inflicted bites having been evi- 
denced clinically, or by the inoculation in the laboratory, and in many 
cases by the death of some other persons or animals bitten by the same 
dogs. No death caused by hydrophobia has been reported among the 
persons inoculated. 

Indigents have been treated free of charge. 

The persons treated were: 
81 from New York. 3 from Missouri. 1 from South Carolina. 



27 


(< 


New Jersey. 


2 


<< 


New Hampshire 


1 


a 


Nebraska. 


16 


u 


Massachusetts. 


2 


a 


Texas. 


1 


a 


Rhode Island. 


11 


a 


Connecticut. 


2 


< . 


Kentucky. 


1 


a 


Arkansas. 


9 


a 


Illinois. 


2 


a 


Ohio. 


1 


a 


Virginia. 


5 


n 


Georgia. 


1 


a 


Maine. 


1 


t< 


Louisiana. 


5 


a 


North Carolina. 


2 


a 


Arizona. 


1 


it 


Indian Ter. 


5 


a 


Pennsylvania. 


1 


a 


Minnesota. 


1 


i i 


Ontario ( Can 


3 


a 


Maryland. 


1 


it 


Iowa. 










With kindest regar 


ds of the Pasteur Institute. 






















Paul Gibier. 



306 THE BACTERIOLOGICAL WORLD. 

OUR SUBSCRIBERS. 

Those who honored us with a trial subscription of three months 
are respectfully reminded that the March number was the last copy to 
which they were entitled. We sincerely trust that the few who have 
neglected to forward the balance of subscription will continue their 
appreciated assistance to the cause of bacteriology in sending as early 
as possible after this issue the sum of $2.25. 



OMISSION. 



Omission in article entitled " Fistula in Ano in Phthisis," by Dr. 
Heneage Gibbes. At page 273, of this number of the Bacteriological 
World, before the last three lines, read : " It is not generally known 
that a sinus exists in the normal contracted state between the external 
and internal sphincters of the anus," and then follow with : "This 
sinus is about one-eighth of an inch," etc., as will be seen on that page. 



AMERICAN MEDICAL ASSOCIATION. 



REDUCED RATES OF RAILROADS. 

We are pleased to give our medical friends the following extract 
from an official notice of the Cleveland, Cincinnati, Chicago and St. 
Louis Railway Company : 

I beg to advise you that a rate of One and One-third Fare for the 
Round Trip, St. Louis to Washington, D. C, has been granted for the 
meeting, on May 5, of the American Medical Association. This rate 
to be made on the certificate plan ; that is, delegates pay full fare 
($19.25) going, and receive from the selling agent a certificate which 
entitles the holder thereof to a ticket for the return passage at one-third 
of the lowest limited first-class rate. 

Tickets will be on sale May 2,3,4 and 5, and certificates for 
return journey will be honored up to, and including three days after 
adjournment of meeting. 

The "Big Four Route" connects in the Grand Central Station, 
Cincinnati, with the picturesque Chesapeake & Ohio Railway, making 
the best and quickest line between St. Louis and Washington. 



THE BACTERIOLOGICAL WORLD. 307 

Leave St. Louis (Union Depot) 8:05 A. M. 7:15 P. M. 

" Cincinnati 6:30 P. M. 7:35 A. M. 

" White Sulphur Springs 5:30 A. M. 8:43 P. M. 

Arrive Washington 2:48 P. M. 6:30 A. M. 

Through sleeping car, St. Louis to Cincinnati, and through sleepers 
and Pullman dining car, Cincinnati to Washington. 

For further information, or sleeping car reservations, address 

W. F. Snyder, 
The Cleveland, Cincinnati, Chicago & St. General Western Agent. 

Louis t Ry. Co. "Big Four Route." 

Broadway & Chestnut, St. Louis, Mo. 



Antiseptic I C ' J I |\ ■ I Non-Toxic 

Prophylactic. J v "— ^ r— 1""^ I \ I P"" Non-Irritant. 

Deodorant. I V J 11 ! \l Non-Escharotic 



LISTERINE 

FORMULA.— Listerine is the essential antiseptic constituent of Thyme, Eucalyptus, 
Baptisia, Gaultheria and Mentha Arvensis, in combination. Each fluid, drachm 
also contains two grains of refined and purified Benzo-boracic Acid. 
DOSE.— Internally : One teaspoonful three or more times a day ( as indicated ) 
either full strength or diluted, as necessary for varied conditions. 
LTSTERINE is a well-proven antiseptic agent — an antizymotic— especially useful 
in the managements catarrhal conditions of the mucous membrane ; adapted to 
internal use, and to make and maintain surgical cleanliness— asepsis— in the treat- 
ment of all parts of the human body, whether by spray, irrigation, atomization or 
simple local application, and therefore characterized by its particular adaptability to 
he field of 

PREVENTIVE MEDICINE-INDIVIDUAL PROPHYLAXIS. 



* LISTEBI1TE * 

Destroys promptly all odors emanating from diseased gums and teeth, and will be 
found of great value when taken internally, in teaspoonful doses, to control the fer- 
mentative eructations of dyspepsia, and to disinfect the mouth, throat and stomach, 
It is a perfect tooth and mouth wash, indispensable for the dental toilet. 
Descriptive literature upon request. 

LAMBERT PHAEMACAL CO., ST. LOUIS, MO. 

AGENCIES: 

S MA"W SON & THOMPSON, ROBERTS & CO., S. PAPPENHEIM, VILANOVA HOS. Y CIA. 

LONDON. E. C. PARIS. BERLIN, W. BARCELONA. 



OLTJB E/ATES ; 



m 




Jill 



< 




\j l 



< 



LD. 



(ILLUSTRATED.) 
$3.00 PER YEAR IN ADVANCE. 



We offer it with the following publications at the rate indicated : 

The Journal 1 American Medical Association 

( WEEKLY.) 
Price, $5.00. With Bacteriological World, $7.00. 



The Journal of Comparative Medicine and Veterinary Archives. 

EDITORS : 

W. A. Conklin, Ph. D., D. V. S., Director Zoological Gardens, N. Y. 

Rush S. Huidekoper, M. D., Veterinarian, Prof. American Veterinary- 
College, New York. 
Price, $3.00. With Bacteriological World, $5.00. 



THE AMEfjICApETERipY REVIEW. 

Editor in Chief: Prof. A. Liautard, Principal, American Veterinary 

College, New York. 
Price, $3.00. With Bacteriological World, $5.00. 



THE CINCINNATI MEDICAL NEWS, 

Editor : J. A. Thacker, A. M., M. D., Cincinnati, Ohio. 
Price, $2.00. With Bacteriological World, $4.50. 



THE MEDICAL HERALD. 

Editor : Daniel Morton, M. D., St. Joseph, Mo. 

Price, $2.00. With Bacteriological World, $4.50. 



ST. LOUIS CLINIQUE. 

Editors : Dr. William Porter and a Corps of Associates, 

St. Louis, Mo. 

Price, $1.00. With Bacteriological World, $3.75. 



Address THE BACTERIOLOGICAL WORLD, Columbia, Mo. 

Or for quicker attention: DR. T. J. TURNER, Mexico, Mo., 

Who has full authority to attend to all business matters for the 
Bacteriological World Company. 



* For the United States, Canada and Mexico only. 




PROF. T. J. BUF^ILL, PH. IS. 



ITU 



Bacteriological World 



A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies. 

Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological World, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to at the 
same office. 

Authors of original articles published in this magazine will be entitled to twenty 
copies of the number contaicing it, provided, that they request the same in writing 
when sending the communication. Reprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. 

Discontinuation : — At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first. of next January. 

collaborators: 

Paul Gibier, M. D., New York. 
Prof. W. T. Belfield, M. D., Chicago. 
Prof. A. W. McAlester, A. M., M. D., 

Missouri University Med. Dep't. 
Prof. Henrt P. Loomis, M. D., 

New York . 



Prof. L. Bremer. M. D., St. Louis, Mo. 
J. W. Stickler, M. D., Orange, N. J. 
Prof. Paul Schweitzer, Ph. D., 

Missouri University. 
Prof. T. J. Burrill, Ph. D., 

Illinois University. 



B. T. Gallowat, B. S., Section of Vegetable Pathology, Washington, D. C. 



U. S. A., MAY, 1891. 



No. 5. 



EDITORIAL. 



CHEMISTRY AND BACTERIOLOGY IN MEDICINE. 

It becomes daily more evident that one no longer can claim 
to be thoroughly equipped to practice medicine without a fair 
knowledge of chemistry and bacteriology. 

Time was — and most of medical colleges seem to think that 
time still is — when chemistry meant merely a glance at this 
science, to give an idea of the combinations of drugs, and their 
incompatibilities. The lectures to students and their very rare 
laboratory exercises, when any were followed, were directed 
chiefly in that direction. Incidentally the chemical constitution 
of the body, its liquids, etc., were touched, and mentioned also 
in connection with physiology. But, at the examinations, chem- 
istry was, and is now in certain colleges, so conducted as to 



310 THE BACTERIOLOGICAL WORLD. 

draw evidence from the students that they know the difference 
between a base and an acid, quinine and morphine, and 
such simple propositions. In many institutions the man who 
knows chemistry fairly well is still an exception. 

Of late years, however, bacteriology has forced a deeper 
study of chemistry on all the progressive and conscientious 
teachers of medicine. Consequently, there is an awakening to 
the fact that medicine was practiced, haphazard, as an art, and 
the truths of it, the sciences underlying it, were ignored. Bac- 
teriology has pointed to various transformations in cultures, and 
in animal and vegetable bodies. It has shown that microbes, 
though causing much disturbance, rarely cause serious troubles 
merely by their presence, but that they do by the chemical sub- 
stauces wrought by their various processes of feeding, at the 
expense of a living organism. Chemistry stepped in and demon- 
strated the nature of the products thus created by parasitic life, 
or by microbes, nourishing themselves at the expense of dead 
matter, such as in cadavers, cheese, cream, meat, oysters, 
etc., etc., or living matter such as the human body. 

It is high time that medical men at the head of colleges 
recognize the fact that a deeper knowledge of the sciences of 
medicine is necessary for the greatest achievement in medical 
teaching and practice. One may follow an empirical rut with- 
out them, but no one can feel that he is doing his duty in 
teaching or practicing medicine in ignorance of the laws 
underlying the very base of physiology and pathology. Dr. 
Vaughan's very able paper on this subject, as read before the 
American Medical Association, at Washington, was timely, and 
should be read by every doctor and medical student in the 
country. 



PROF. BURRILL. 



We take a great deal of pleasure in presenting to our read- 
ers in this issue the picture of one of the oldest and most 
favorably known of the bacteriologists of the country. Indeed, 
Dr. Burrill is one of the foremost bacteriologists in the world, 
and his word *on technology is authoritative everywhere. In the 
study of tuberculosis he rendered a very great service to all 



THE BACTERIOLOGICAL WORLD. 311 

interested in that line of investigation, and especially to prac- 
ticing physicians, by his clever combination of a simple method 
of staining the bacillus of tuberculosis. At the time this was 
done bacteriological technique was very crude. There were no 
simple methods nor any easy ones, and BurrilPs stain was no 
little discovery. He has done much work in the bacteriology 
of plant diseases as well as the diseases of animal life. Science 
to-day owes much to his brains, energy and patience in working 
intricate problems. We regret much that we have not a list of 
the numerous literary works and deeds of valor of this advanced 
scientific thinker. 



312 THE BACTERIOLOGICAL WORLD. 

LESSONS IN BACTERIOLOGY. 



LESSON FIFTH.— PART I. 



BY PAUL PAQUIN, M. D. 



CULTIVATION OF BACTERIA — STERILIZATION. 

Good results can be obtained in bacteriological studies only 
on condition of absolute cleanliness in the laboratory and field 
manipulations. In the first place, bacteria cannot be studied 
with profit if mingled with numerous organisms. One can- 
not clearly understand the morphology or physiology of so 
minute microscopic beings in a field of observation in which 
different living particles strive for existence. 

The first step then is the isolation of the microbes, i. e. 
the separation of the different kinds in order to insure the study 
of each independently and in a state of purity. This isolation 
is effected by means of the cultivation of the organisms in vari- 
ous suitable media, the composition of some of which we have 
already seen. 

The isolation of microbes is an extremely difficult task, 
and requires patience, integrity and a love of science. It 
means that every step, every manipulation, however slight, 
must be based on the greatest caution possible against all forms 
of contamination. The air, we might say, is simply full of 
germs, and could we but see the countless living organisms 
floating in the space in our laboratories or offices, for instance, 
they would appear as a dense cloud impervious to our vision. 
Every object, then, every vessel, knife or other instrument 
is the bearer of microbes, and must be freed before being 
used. In other words, these objects must be sterile, free from 
living forms of any kind. 

We understand, then, that a reliable study of bacteria 
depends first on their cultivation and isolation, and that this 
can be successful only under the strictest sterilization methods. 

Let us now go into details as to the various sterilization 

questions involved. 

The room. — The apartment in which cultures and isola- 
tions of microbes are made should be as free from dust and 



THE BACTERIOLOGICAL WORLD. 313 

floating bacteria as possible. This is better attained by using a 
very small room with perfectly smooth walls and ceilings, free 
from cornices, nooks, cavities, window sills, or anything else 
capable of catching dust and germs A room, say 6x6, with 
glossy walls, floors and ceilings, glazed or painted in such a 
smooth manner, and with such material as to admit of perfect 
cleanliness and disinfection, provided with only one light, is in 
my judgment the very best place to make cultures. Such a 
place may be shut air tight almost at night, a sulphur brick 
burned in the center of it, and thus sulphurous acid fumes be 
generated for several minutes, and the atmosphere of the room 
be much purified by contact with sulphurous acid gas for sev- 
eral hours. This room may also be purified by steam, or again 
it may be disinfected, rendered almost sterile by spraying 
a bi-chlonde of mercury solution, 1 to 2,000, through the 
space in the evening, closing the room till the next morning 
when it may safely be entered. 

The culture room should be kept sacred, and no visitors 
admitted. The oftener it is opened the more its atmosphere 
becomes polluted. 

The operator should be clothed with a clean jacket, say a 
white tough cotton cloth coat, and overalls of same material; 
the sleeves might be protected by oilcloth sleevelets, such as 
grocers use sometimes. These clothes should be removed and 
left in the culture room every night when sterilization of the 
latter occurs, and they should be washed every week at least. 
The hands and hair should be well cleansed, so nothing therefrom 
shall contaminate instruments, etc. It is better, in fact, before 
doing anything in cultures and during the processes, to dip the 
hands occasionally in a solution of bi-chloride of mercury, 1 to 
2,000, say, kept in a convenient place for the purpose. Kings 
should be removed from the fingers before putting the hands 
in this solution; otherwise they may be seriously damaged. 

The instruments. — Of all the sterilization agencies heat 
is undoubtedly the most effective. But unfortunately it cannot 
always be applied with safety or impunity. Now it would be 
very difficult to sterilize a room, however small, by heat, unless it 
be steam, and even this is costly. Instruments, however, can 
most all be sterilized by heat of some kind. Heat as a 
sterilizer is applied either moist or dry, i. e., hot water, steam, 



314 THE BACTERIOLOGICAL WORLD. 

dry air or actual fire (flame, cauterization). It is important to 
utilize the least destructive of these kinds of heat in sterilizing 
instruments of various kinds. 

Good glass instruments may stand either moist or dry heat, 
including a blaze, but a knife, a scalpel, a pair of brass or 
steel forceps would be ruined if kept in a blaze until a change 
of color occurs. A mercurial preparation is not injurious to 
glass goods, wooden ware, horny substances, etc., but is bad on 
the edge of a cutting instrument. 

To sterilize instruments then it is desirable for the reasons 
above given, and many others too long to explain, to observe the 
following rules closely : 

First. — Sterilize empty glassware at dry heat. 

Second. — Sterilize glassware filled with culture media or 
other substances in moist heat. 

Third. — You may sterilize the surface of knife blades, pin- 
cers, forceps or other iron, steel, brass instruments by passing 
them slowly in a gas blaze to kill the organisms on their surface, 
but never heat them long, or until steel blackness, dark red or 
whiteness is produced. 

Fourth. — Platinum instruments, such as forceps with plati- 
num points or inoculating needles, may be sterilized by heating 
white. 

Fifth. — Cotton, wool, paper and like things should be 
sterilized only in dry hot air. 

Sixth. — Clothes, towels, rags, handkerchiefs, etc., may be 
sterilized either in dry air or by steam. 

Sterilization of cultures and organic materials. 
These should be sterilized usually by steam, though such as 
blood serum is sterilized by low dry heat. The details of such 
sterilizations will be given in speaking of the substances them- 
selves. 

The degree of heat in various sterilizations, whether they 
be dry or moist, is another important item. Now 212° F., as 
explained to you ere this, is usually high enough to destroy most 
of micro-organisms, in a very short time, both in dry and moist 
heat. But sometimes it is impracticable to raise the tempera- 
ture that high and again it can readily and safely be raised very 
much higher. For instance, a beef-broth culture with gelatine 
sterilized once a day about 212° F, not higher, for say five or 



THE BACTERIOLOGICAL WORLD. 315 

six consecutive days, will be prettier, clearer, and usually remain 
solid in warm temperature, whilst the same material sterilized 
at one heating over 212° will present often the reverse condi- 
tions ; it will not remain solid in warm weather. 

Again, blood serum, sterilized gradually at a low heat, will 
finally be free from germs and yet remain comparatively clear 
and transparent, whilst the same at high temperature will at once 
become white and opaque and unsuitable for most of cultures. 

The apparatus for sterilization are now very numerous and 
many of them are costly, too costly in fact for daily use except 
in laboratories. The busy practitioner or the amateur, however, 
may readily sterilize most of things in the office or kitchen. 
The kitchen stove oven can well serve as a dry heat sterilizer, 
and the steam kettle or boiler as a moist heat (boiling water, 
water bath or steam) sterilizer. I have had occasion to try 
these and succeeded fairly well by repeated heating at tempera- 
ture lower even than 212° F. 

The special laboratory sterilizers are as follows : 

A dry heat oven, consisting of a double wall metallic box, 
(sheet iron), about eight inches wide, six or seven inches 
deep and eighteen inches high, inside measure. A little open- 
ing at the top admits of a thermometer.* 

Koch's vapor sterilizer : This instrument permits the 
raising of temperature very high, say about 324° F., thus 
destroying all organisms and their spores readily in a few 
minutes. Ohamberland's autoclave is another sterilizer of the 
same kind. There is no difference in these high heat sterilizers 
except in their mechanical construction and convenience. All 
of them give the same results. 

Besides these methods of sterilization in laboratory tech- 
nique there is another now often followed in sterilizing certain 
liquids. I mean the Chamberland filter. The essentials of 
this apparatus consist of a hollow cylinder bougie of porcelain, 
closed at the bottom and provided with a smooth funnel-shaped 
point at the other extremity, over which may be adapted a piece 
of rubber tube. By a system of pressure or suction the liquid 
passing through this bougie is freed from micro-organisms. 
(This lesson to be continued with figures in June number.) 

* By a misfortune the cuts to illustrate the instruments of this 
lesson have not been received, and will be used only in the next issue. 



316 THE BACTERIOLOGICAL WORLD. 

SURGICAL BACTERIOLOGY. 



SURGICAL TREATMENT OF PULMONARY CAVITIES. 



The bold endeavor to radically attack phthisical cavities 
and drain them, although introduced some years ago, has been 
lately receiving fresh advocacy, especially in relation to the 
treatment of phthisis by Koch's "tuberculin." Many of the 
dangers of this remedy in advanced cases of phthisis rest upon 
the retention of necrotic material within the lung, and the 
liability to its being inhaled into previously unaffected areas. 
Dr. Sonnenberg recently operated upon four cases undergoing 
this treatment, and Dr. E. Leser, of Halle, reports two more. 
One of these was a female aged 43, who had had symptoms for 
four years, and presented physical signs of excavation at the left 
apex. The operation consisted in making an incision in the 
first intercostal space, 7 centimetres long, and, after dividing 
the muscles and securing blood-vessels, introducing a trocar in 
the direction of the cavity. Although no fluid could be 
obtained, it was evident that the point of the trocar was in the 
cavity, and the pleurae being firmly adherent an aperture was 
bored through the layer of tissue overlying the cavity by means 
of a Paquelin's cautery, the trocar being taken as a guide. 
The cavity was the size of an apple, had somewhat irregular 
walls, and was coated with flakes of inspissated pus and caseous 
matter; these were removed, and the cavity stuffed with anti- 
septic gauze. Subsequently, the patient was injected with 
Koch's "tuberculin/' which always caused increased discharge 
from the cavity; and later still some of the fluid was directly 
introduced into the cavity itself. At the last report the patient 
had improved much in general health, and the cavity had 
dwindled to one-half its former size. The second case, a man 
aged 42, was similar, except that in him the cavity was no 
larger than a chestnut, and was situated at the right apex. 
Here, too, the use of tuberculin produced increased discharge, 
and on one occasion its injection into the cavity was followed 
by severe vomiting, dyspnoea, and other alarming symptoms. 



THE BACTERIOLOGICAL WORLD. 317 

Dr. Leser, however, thinks this mode of treatment has a future 
before it. At present he would make it a sine qua non that 
the costal and pulmonary pleurse should be firmly united, as 
they usually are when cavities are of some standing, and he 
enters fully into the technique of the operation, laying stress 
upon having a fairly large incision through the thoracic Wall, 
but not a very large aperture made in the lung, since this will 
be sure to increase by sloughing. The notable feature of these 
cases seems to be the immediate relief to cough and expectora- 
tion given by the external opening, although, when the injec- 
tions are resorted to, the secretion becomes so much more 
abundant. — The Lancet. 



HYGIENIC BACTERIOLOGY. 



THE BACILLI OF MALARIA. 

Dr. xindreas has recently published an interesting treatise 
on the comparative number of the bacilli of malaria in the air 
at different times of the day. His experiments, which were 
conducted in the Observatory of Moncalieri, and reported in 
the Medicinische Neuigkeitten, April 11, 1891, were carried out 
by means of small rubber balloons filled with hydrogen. On 
to these balloons he fastened a small box, holding prepared 
glass slides, which box he was able to open by means of a cord, 
after the balloon had reached the desired height. Microscopi- 
cal examination of the slides showed that in the early hours of 
the day the swarms of bacteria were close to the ground and in 
large numbers ; later, at about nine o'clock in the morning 
until about three in the afternoon, they would rise until they 
reached a considerable height, and from that time would again 
gradually sink to the ground. The number of bacilli in the 
air was almost exactly in proportion to the rise of temperature, 
while in direct opposition to the amount of humidity in the 
atmosphere. It is evident, therefore, that the condensation of 
the watery vapors in the air cause the falling of the bacilli, and 
for this reason the morning and evening hours are the most dan- 
gerous in malarial districts. — Medical and Surgical Reporter. 



318 THE BACTERIOLOGICAL WORLD. 

INFECTION THROUGH THE SOIL. 

The question is often raised whether there is any direct or 
indirect proof of the infectivity of the bodies of those dying 
from specific fevers, or whether the soil of graveyards may 
become really dangerously contaminated thereby; but it is also 
one to which comparatively little attention has been given by 
the experimentalist. One aspect of the matter has been lately 
studied by Dr. Justyn Karlinski, of Konjica, Herzegovina ; in 
a series of experiments (Centralblatt f. Bakteriol, ix, 13) 
undertaken to determine whether the organs of the body 
undergo any change in temperature during the natural process 
of decomposition after burial in the earth, and especially 
whether any differences in this regard are to be met with in the 
case of infected subjects. His results show that invariably the 
putrefactive process is accompanied by a rise of temperature 
above that of the circumjacent soil ; and also that this rise in 
temperature is markedly higher when the parts examined have 
been taken from the bodies of men or animals who have 
succumbed to infective diseases than it is in the case of healthy 
bodies. He further examined into the question of the survival 
of the pathogenic bacteria in the affected parts, and states that 
the typhoid bacilli may retain their vitality in the decomposing 
spleen for three months, and are only completely annihilated 
by rapid putrefaction and the presence of a large number of 
the bacteria of putrefaction. Dr. Karlmski says that he had 
previously shown that typhoid bacilli could retain their vitality 
for a period of five months in soil, but that if the earth were 
thoroughly saturated with rain water they were destroyed in 
from seven to fourteen days. The part played by the soil in 
the origin of epidemics should not, he thinks, be underesti- 
mated, since typhoid bacilli can only exist in water a compara- 
tively short time.— Lancet. — Medical and Surgical Reporter. 



INFECTION FROM MILK. 

In the Glasgoiu Medical Journal, for October, appears an 
account of an epidemic of erysipelas and sore throat, occurring 
among families supplied with milk from a certain farm. The 



THE BACTERIOLOGICAL WORLD. 319 

most striking symptom was an intense inflammation of the 
fauces, resembling erysipelas of the mucous membrane, with 
swelling of the glands of the neck, and in some cases suppura- 
tion. In some, true erysipelas of the skin developed. The 
temperatures ranged from 102° to 105° F. during the first few 
days of an attack. Convalescence was attended by extreme 
prostration. No bacterial examination was made, but a clear 
connection was traced between the milk and the epidemic. — 
Journal American Medical Association. 



TYPHOID AND PSEUDO-TYPHOID BACILLI IN RIVER 

WATER. 

Dr. Cassedebat (Annates cle V Institnt Pasteur), following 
up Rietsch's work on the river waters of Marseilles, gives the 
result of an examination of seventy specimens of water from 
which 250 cultivations were made, with the view of deter- 
mining whether the Eberth bacillus is to be found in the 
waters of the Durance which supply a part of the city, where 
typhoid fever is endemic and often epidemic. In no case was 
he successful in finding the Ebertb bacillus, but he was able 
to separate three bacilli — " pseudo-typhoid bacilli" — which 
resemble the typhoid bacilli in many respects. The plate culti-. 
vations are so much alike that unless all four can be seen 
together it is difficult to distinguish one from the other. None 
of the pseudo-forms are quite so toxic to white mice as the true 
form, and one of them is quite innocuous. Although Casse- 
debat was not able to find the true form in water taken from 
the water supply which was most open to contamination, he 
found that this was not because the bacilli could not live in 
water, as in distilled water to which a cultivation had been 
purposely added, he could easily distinguish its presence at the 
end of forty- four days, and, when added along with half a dozen 
other forms, he could find them at the end of seventeen days. 
He comes to the conclusion, therefore, that the true typhoid 
bacillus does not occur in water so frequently as is sometimes 
represented, and that one or other of the forms of pseudo- 
typhoid bacilli has in certain cases been mistaken for it. — 
Exchange. 



320 THE BACTERIOLOGICAL WORLD. 

ORIGINAL ARTICLES. 



EXPERIMENTAL RESEARCH ON PROFESSOR R. KOCH'S 

FLUID, WITH EXHIBITION OF CULTURES 

AND EFFECTS UPON ANIMALS '.* 



BY PAUL GIBIER, M. D. 

Director of the New York Bacteriological Institute . 
Having received a certain quantity of Dr. Koch's ' 'lymph" in 
the beginning of January, I made several experiments with it, 
with the results of which I should like briefly to acquaint you. 
These experiments were made : 

A. Upon the cultures of tuberculosis. 

B. Upon animals. 

C. Upon patients. 

A. Experiments upon cultures. — 1. A milligram of 
" lymph" is added to five cubic centimetres of glycerin-agar, 
which is then inoculated with a liquid culture of human tuber- 
culosis. 

Result. — After fifteen days' sojourn in the incubator the 
whole surface of the agar was covered by a culture of tubercle 
bacilli. 

2. Five milligrams of ( ' lymph " were added to five cubic 
centimetres of glycerin -agar and inoculated as in the pre- 
ceding experiment. 

Result.— Fifteen days later the growth, although less 
abundant than in the first case, covered all the surface of the 
culture. 

3. Fifteen cubic millimetres of the culture of the tubercle 
bacilli were mixed with an equal quantity of pure "lymph," 
and glycerin-agar was inoculated with it at the varying inter- 
vals of a quarter of an hour, and of one, two, nine, twelve and 
twenty-six hours. After a contact of twenty-six hours the 
cultures developed normally, as you can see by the tubes which 
I have the honor of presenting for your inspection. The bacilli 
themselves having sojourned for this length of time in contact 

* Read before the Medical Society of the County of New York, 
February 23, 1891. 



THE BACTERIOLOGICAL WORLD. 



321 



with the " lymph" do not seem, on microscopic examination, 
to have been perceptibly modified. 

B. Experiments upon animals. — I will only, among 
several, quote the following : 1. A guinea-pig weighing 774 
grams inoculated with a cubic centimetre of culture of human 
tuberculosis, sterilized by gold cyanide, forms a large abscess 
after a few days, which extends over the greater part of the 
right thoracic and abdominal surfaces. This is found to con- 
tain an abundant quantity of caseous pus, and, while pure, 
numerous dead bacilli. The abscess healed without entailing 
skin sphacelus, and the animal increased in weight. His actual 
weight is 811 grams, giving an increase of 37 grams. 

Several milligrams of " lymph," injected into this 
guinea-pig at different epochs, produced no reaction, even in 
the dose of eighty milligrams. 

2. A guinea-pig inoculated November 22, 1890, with 
human tuberculosis by means of two cutaneous incisions in the 
groins, presented a slowly evolving tuberculosis. The inguinal 
nodes, at the end of three weeks, were as large as small nuts ; 
the animal, while apparently well, had lost 40 grams, weigh- 
ing 763 grams January 2. He was then given half a milli- 
gram of "lymph," and the successive days the following 
quantities : 



January 3, 


% 


milligram. 




January 27, 


15 


milligrams. 


< i 


4, 


1 




a 




( i 


30, 


15 




1 1 


.< 


5, 


1 . 




tt 




February 2, 


30 




c < 


a 


6, 


1% milligrams. 




i . 


7, 


20 




a 


i < 


8, 


3 




< t 




i i 


9, 


25 




,i 




10, 
11, 


3 
3 




. t 




1 ( 


10, 
12, 


25 
30 




a 


a 


12, 


3 




I i 




i > 


14, 


35 




i t 


it 


13, 


3 




i . 




• i 


16, 


75 




it 




14, 
16, 


5 

10 




i i 






18, 
20, 


80 
100 




1 1 
1 1 


i t 


18, 


10 




I i 




i. 


22, 


100 




1 1 


a 


20, 


15 




I i 














. t 
t t 


22, 
>5, 


15 
15 




. 1 
it 




Total 




640 


milligrams. 


Makin 


g a 


total 


of 640 


millis 


'rams in a 


little less 


than two 



months' time. 



322 THE BACTERIOLOGICAL WORLD. 

A reaction was only obtained after the injection of 30 mil- 
ligrams. I should like here to remark that, given the weight 
of the .animal, ] 00 milligrams of lymph would be equivalent 
to about ten grams, or 10,000 milligrams, for a man of 
ordinary weight, and the total quantity of 640 milligrams, 
equivalent to more that 64 grams or cubic centimetres, for 
the same man. The reaction, obtained after the injection of 
30 milligrams and larger doses, manifested itself by an ele- 
vation of one or two degrees Centigrade of temperature. Neither 
the tubercular ulcerations nor the nodes were in any way 
affected. 

Results. — They are absolutely unfavorable ; the animal has 
lost weight constantly, weighing at the beginning of the exper- 
mentation 763 grams, and to-day 645 grams, giving a loss 
of 118 grams, or one seventh of its entire weight, since the 
beginning of the injections. Moreover, as you can see, the cuta- 
neous lesions of the animal, which I present you, have acquired 
an unusual intensity. The points of inocculation, which 
had cicatrized, reopened during the period of injections and 
became ulcerated, forming on the right side a raw surface of the 
size of a twenty-five cent piece. The ulceration of the left side 
is much larger and extends from the flank toward the back and 
around the thigh, and invades the abdominal surface. Other 
smaller ulcerations and tubercular nodules can be seen at a 
distance of a centimetre and a half from the main wound. 
Eare bacilli have been found in the tissue excised from the 
edges of this ulceration. Localized induration of the skin is 
found at about two centimetres and a half from the ulceration, 
and it is positive that a bacillar infiltration has taken place. 
The abdominal wall is infiltrated. The internal organs are 
apparently compromised, for respiration is notably accelerated, 
there being about ninety-six inspirations to the minute, and the 
animal having apparently but a short, time to live. I intend to 
exhibit its viscera to the society whenever it dies. 

I do not believe that this case can be considered excep- 
tional, for similar results have been obtained and communicated 
to the Academy of Medicine, of Paris, by Professor Jaccoud, 
Professor Dujardin Beaumetz and Professor Dubief. 



THE BACTERIOLOGICAL WORLD. 323 

C. Experiments upon patients. — In consequence of 
the scarcely favorable results which have been published so far, 
I have not felt myself at liberty to accept more than three 
patients for the inoculations. Two of these patients have 
pulmonary tuberculosis capable of amelioration by the ordinary 
methods of treatment ; the third patient, who has been 
operated upon several times in the past two years for tubercular 
genital lesions, had at the moment he presented himself for 
inoculations a tubercular lesion of the condyles and lower third 
of the left femur. Although the articulation is comparatively 
mobile, the leg is several centimetres shorter than it should be, 
and complete extension cannot be obtained ; walking is only 
possible by means of crutches, and the slightest contact is 
painful. 

The condition of the two patients with pulmonary tuber- 
culosis has not been visibly ameliorated by the injections. The 
third patient with tubercular bone affection has seen his pain 
disappear so that neither pressure nor blows upon the diseased 
parts is any longer painful. 

He has increased over four kilograms ( nearly ten 
pounds ) in weight in five weeks, his weight at the beginning 
of the treatment being 63,6 kilograms. He is always 
forced to use crutches, although able to lean upon the foot of 
the affected side, a thing he could not do less than a month 
and a half ago. I have believed myself warranted, in view of 
this condition of affairs, in continuing the treatment by injec- 
tions, although I have deemed it advisable to add to it an 
iodine medication and a special diet. 

Conclusion A, The addition of a milligram of " lymph" 
to a cubic centimetre of culture does not prevent the develop- 
ment of the tubercle bacilli. 

Tubercle bacilli can develop after an interval of several 
hours ( twenty-six hours at least ) of contact with pure 
" lymph. " 

Contrary to what would appear from certain recorded 
cases, even the direct action of the lymph upon the tubercle 
bacilli does not produce any appreciable morphological or bio- 
logical modification. 



324 THE BACTERIOLOGICAL WORLD. 

B. The injection of the " lymph" of Professor Koch, 
contrary to his assertions at the Berlin Congress, and in his 
subsequent communications, exercised no inhibitory action, 
even in considerable doses, upon the development of the but 
slightly advanced tuberculosis of the guinea-pig in the above- 
mentioned experiment. 

0. The history of the patients of whom I have spoken, as 
well as the careful study of the observations published, war- 
rants my opinion that in other than cases of tubercular lupus 
or of specific bone lesion the " tuberculin" of Koch is ineffi- 
cacious. 

Taking into consideration results similar to those I have 
described, we could conclude that we are, for the moment, 
assisting at the spectacle of one of the greatest medical and 
scientific delusions that has ever existed, and that the remedy 
in question should be completely rejected from therapeutics. 
Still, the reaction which takes place, according to many case 
records, at the diseased foci, the favorable results obtained in 
certain special cases ( lupus, tuberculosis of the larynx, of the 
bones, etc.) permit us to believe that the new remedy could be 
of use not only as a method of diagnostic but also as a precious 
adjuvant to other methods of treatment of tuberculosis. This 
is a view of the case which, in so far as I know, has been as yet 
but little studied, and which, it seems to me, merits considera- 
tion. 



STERILIZED MILK AS INFANT FOOD. 



Read before the Missouri State Medical Association, Excelsior Springs, 

Mo., May 20, 1891. 

BY PAUL PAQUIN, M. D. 

The heavy mortality of bottle-fed babies owing to the 
unsuitableness of the food which they are forced to ingest, and 
the meagerness of their development in many instances when 
they do resist the strain of unnatural food on their delicate ali- 
mentary canal, has been noted and deplored by the civilized 
nations a very long time. The medical world has long been 



THE BACTERIOLOGICAL WORLD. 325 

searching for means to effectually remedy the evil. The. chem- 
ists, bacteriologists, have been for many years experimenting 
with a view to overcome the disaster among the babes and 
young children. As a result we find our pharmaceutical mar- 
kets now filled with various artificial foods, all with claims 
of superiority on the other. 

Some of these foods contain absolutely no milk, nor any 
true elements of the milk. Others are formed of milk partly, 
with the addition of other ingredients ; such is condensed milk. 
Others again contain milk elements in the form of a powder. 
Still others are the result of a process of evaporation of milk. 
And y finally, we have a number of more or less valuable or more 
or less unwise medico-dietetic formulas with or without milk. 

All this comes from the quandary in which the parents and 
physicians are placed when they are confronted with an infant 
so unfortunate as to be forbidden, by some cause or other, the 
use of a mother's milk. 

These various foods, however, whether containing milk or 
not, except in one instance of a sterilized evaporated cream 
which I will mention further, have not succeeded in replacing 
the mother's milk, at least in the very young, though all of 
them have more or less value as foods at certain ages and in. 
certain conditions. For a very young child, there is as yet no 
absolute substitute found for healthful human milk so far as 
its digestibility in a human stomach is concerned. The nearest 
to it is cow's milk, though many pretentious opinions have been 
given on the subject which tend to depreciate its value as infant 
food. 

In comparing the composition of cow's milk and of human 
milk we find (in taking Doyere's analysis as the average of a 
number of analyses of authoritative experts) the following ele- 
ments and proportions: 

Water. Butter. Casein. Albumen. , Sugar. Salts. 

Woman 87.38 3.80 0.34 1.30 7.00 0.18 

Cow 87.60 2.20 3.00 1.20 4.70 0.70 

We see that the average cow's milk contains $.66 per cent. 
more casein than the human milk, 1.60 per cent, less butter, 
2.30 per cent, less sugar. The other differences of composition 
are comparatively unimportant. 



326 THE BACTERIOLOGICAL WORLD. 

A baby's digestive organs, then, are by nature prepared to 
digest at the outset only 0.34 parts of casein in each quantity of 
milk sucked from the breast. If it is fed cow's milk it receives 
at once, without its stomach being prepared by a gradual aug- 
mentation, 3 parts of casein or 2.66 per cent, more than it was 
prepared to or supposed to be capable of digesting properly 
immediately after birth. 

But these differences in the constitution of the milk are 
not the only causes of disturbance in the digestion of infants. 
We must consider also that breast-raised babies get every two 
or three hours milk warmed by nature's process in the mother's 
breast, absolutely free from any of the ferment-producing organ- 
isms which cause the curdling, souring and decomposition of 
milk, whilst bottle fed babies, on the contrary, get cow's milk 
warmed by artificial heat to such a degree, sometimes, that the 
casein, already difficult of digestion because of its excess, is 
coagulated and toughened, and, from the moment of milking 
to the hours of feeding, it receives germs from the udder, the 
hands, the cow's hair and skin, and too often manure and urine 
even, not counting the innumerable air micro-organisms which 
fall constantly into the fluid. 

Thus we have three grave objections to a true substitution 
in every particular of cow's milk for human milk : First, the 
differences in the constitution; second, the differences between 
naturally and artificiallg warmed milk ; third, the differences 
between a milk free from the contamination of ferment organ, 
isms and that of the milk wherein they may exist or may have 
existed and caused transformations. It would seem then that 
it were a simple thing to efface most of these differences and 
bring cow's milk very close to the composition and nature of 
human milk. Not so, however. What in theory appears feas- 
ible is practically beyond perfection with our present means and 
knowledge. 

Still we may effect some intensely useful and practical 
corrections, bringing the two kinds of milk very close in their 
physiological effects on the system of the infant, so much so 
indeed that, by the use of improved cow's milk, in foundling 
asylums, where the tender innocents generated by heartless 
sensuality, thrown to the mercy of selfish and cruel humanity, 



THE BACTERIOLOGICAL WORLD. 327 

never know the loving care of a mother, and where the ratio of 
death was from 75 per cent, to the appalling number of 100 per 
cent, in summer, the death rate has been reduced to 25 per- 
cent. These figures were given. by physicians in charge of such 
institutions in large eastern cities. Now how are we to effect 
these changes and save the babies in the homes and the 
asylums? Let us examine into the problem. 

First as to the constitution : Here we should add fat, 
extract casein, and add sugar. Unfortunately, it seems at 
this date that the best we can do is to add a little water and 
sugar, and trust to nature to become competent to digest cow's 
milk, after gradual preparation of the infant's stomach and 
sterilization of the milk. More further on this point. 

Second as to the warmth : Milk just from the breast or the 
udder seems to have a vital property not well determined. 
The fresh, warm product of living cells is, we would infer, a 
nourishment containing vital energies to be found nowhere else 
in nature. As it flows from the breast of a healthy mother 
into the infant's stomach, it passes directly from life to life 
without any interference or influence playing on its nature. 

Now what occurs in cow's milk warmed artificially ? If it 
does contain a vital principle the cooling process and the heat- 
ing again, between the periods of milking and the periods of 
feeding, doubtless devitalize it, if I may so speak, particularly 
if a high temperature is used. I must admit, however, that 
this conclusion is theoretical, and inferred from physiological 
laws. 

Third, as to the contamination ivith micro-organisms : 
This is by far the most important and most complicated ques- 
tion of the three. 

I need not go into a lengthy discussion to prove that the 
transformation of milk is due to the growth of micro-organisms. * 
It is a recognized fact that the souring and curdling of it, as 
occurs daily in the ordinary course of things, are due to micro- 
scopic living beings. The effect of ozone in a thunder storm 
seems to hasten the souring, but this particular result is an 

* A French author recently advanced the theory that the micro- 
organisms in question are animalcules, and that souring, etc., is not 
due to bacteria proper. 



328 THE BACTERIOLOGICAL WORLD. 

exception and seemingly a chemical action, which has no bear- 
ing at any other time possibly than in electrical storms. The 
soured condition is merely the condition of milk m which lactic 
acid has been formed from the sugar it contained, by the feed- 
ing and digestive processes of germs. The coagulation or 
curdling is likewise the result of germ life. 

The process of the transformation of milk after milking 
is as follows : 

First, the production of lactic acid by the micro-organism, 
commonly known as the lactic acid ferment, which develops 
and feeds itself at the expense of constituents of the milk. 
This acid then causes coagulation. As the milk grows older, 
other micro organisms feed upon the coagulated portions, as 
well as the fluids, and produce other transformations with the 
natural results of chemical products, which I do not intend to 
discuss here. 

I need not say that milk so affected by micro-organisms is 
injurious to the young. You all have seen ill effects of feeding 
cow's milk in various conditions, ages of the subject and 
atmospheric temperatures. You all know that lactic acid, 
butyric acid or other products of milk fermentations (or decom- 
positions) are not well tolerated by -the stomach of the infant, 
and that various more or less disastrous results follow, par- 
ticularly during warm seasons, when the germs develop more 
quickly. 

But when we compare these results in the young with 
those of feeding similar milk in the older children and the 
adult we, at first, fail to comprehend why there should be such 
a difference. In my judgment the matter is readily explained 
by the difference in the process of digestion between the suck- 
ling infant and the older children or adults eating and masti- 
cating food, and the action of micro-organisms on the milk. 

In the first place, the stomach of a babe does not contain 
muriatic acid, whilst that of the masticating child or adult 
person does. Therefore, the stomach of the former is not pre- 
pared to stand the irritant touch of lactic acid, which is formed 
before or after entering it by the action of the germs that con- 
taminate the cow's milk. In the masticating human being the 



THE BACTERIOLOGICAL WORLD. 329 

9 

stomach contains muriatic acid, and is capable of receiving 
such irritant agents. 

Again, in the babe, the muriatic acid which is a fine 
destroyer of many forms of microbes being wanting, all germs, 
not excluding the souring and curdling ferments which have 
entered the digestive tract with the cow's milk, grow uninter- 
rupted. They vegetate, multiply with wonderful rapidity, pro- 
ducing thereby irritant poisonous ptomaines perhaps, gas, or 
other bacterial products throughout the alimentary canal, and 
the result is, diarrhea, cholera infantum, gaseous colic and 
other more or less dangerous disturbances. In the stomach 
which is old enough to secrete muriatic acid, on the contrary, 
the many germs ingested meet first this acid, which may in 
part arrest them in or retard their vegetation. But, should any 
escape and fermentation follow in the stomach and intestines, 
or should a masticating human being swallow milk- already 
altered in various ways by microbes, the stomach and bowels are 
usually found accustomed to the digestion and partial fermenta- 
tion or decomposition of various foods, and are prepared for and 
capable of resisting the irritant and disturbing properties of 
bacterial and ferment products. 

Perhaps, we will realize these facts better in bringing the 
digestive phenomena of milk in the infant and adult to our 
minds. 

In the suckling infant, the comparatively small proportion 
of coagulable matter of the human milk, or the great amount 
in cow's milk, coagulates in the stomach by the action of the 
so-called rennet ferment, comparable to the rennet in calves. 
In the adult, coagulation occurs in the stomach also, but by the 
acid of the gastric juice instead of the special ferment principle 
of the stomach of infants. So coagulation of milk in all human 
beings occurs in the stomach, but by the action of different 
principles. In neither case, however, does the coagulum formed 
redissolve in the stomach. The particles of coagulated casein 
pass on into the intestines and there meet the intestinal juice 
which digests them. The ferment capable of dissolving casein 
curds seems to exist in the pancreatic secretion. Thus, the 
infant's stomach, prepared to receive only about 3-10 of casein 



330 THE BACTERIOLOGICAL WORLD. 

in each quantity of milk swallowed, can well dispose of this 
minute proportion even if it were ingested after infection by 
various microbes, but when it comes to pour into it cow's milk 
containing an average of 3 parts of casein for each portion, and 
full of ferment germs and decomposing bacteria, we at once 
produce an abnormal amount of clotted milk in the stomach, 
which in itself sickens the babe. Then this extraordinary 
amount of casein passes into the intestines, and there fails of 
digestion because of the deficiency of pancreatic secretion in the 
infant. What occurs in such circumstances ? Why the casein 
is attacked by various micro-organisms, decomposed by them, 
and various irritant products of a gaseous liquid or solid char- 
acter are produced which cause disease. 

It seems evident, then, that to the action of micro-organ- 
isms upon milk and to the surplus of casein in cow's milk we 
must attribute the severe diseases of the digestive organs of 
bottle-fed babies. These infinitely small, these microscopic 
plants, we must charge chiefly with the appalling death rate 
among children reared on cow's milk. 

To remedy the evil, as I have said already, a great number 
of artificial foods, and various processes of preparing or rearing 
infants deprived of a mother's breast milk have been recom- 
mended. Perhaps the first and oldest means tried to substitute 
mother's food was the wet-nurse, and to this day she is recognized 
in many cases as the only means of saving a recently born child's 
life. Indeed, the very first days or weeks, it is doubtful if any 
mode of procedure can take tier place if she be of the right 
constitution, temperament, and in good health. But, unfor- 
tunately, the circumstances attendant to a wet nurse's life are 
almost always seriously objectionable from many standpoints. 
It may be from a moral point of view. There are nearly always 
either some moral or physiological or pathological objections. 
Sometimes all these exist. 

In the first place, the woman who lends herself to such 
services, if she is honest, has very frequently a heavy sorrow on 
her heart which is incompatible with the best nutrition and the 
secretion of the healthiest of milk. In the second place if she 
is indifferent in her condition she may present dangerous 
constitutional defects produced by a life of shame or more or 



THE BACTERIOLOGICAL WORLD. 331 

less debauchery. It the third place there is always the dangers 
of occult tuberculosis, scrofulotic and syphilitic taints. In the 
fourth place their fees are beyond modest means. Fifth, most 
of them become tyrannical; in their knowledge that once their 
task begun, even at an exorbitant price, they hold the mother 
and father in their clutches for many months, they often 
become most disagreeable persons to deal with. And finally 
the death rate or ill health of children so reared is not much 
below that of careful artificial feeding by a good mother and an 
intelligent physician. 

All the artificial foods you know well. All are more or less 
nutritious and often constitute excellent accessories to the diet 
of a child or an invalid, but none can be used exclusively for 
baby food. The evaporated cream termed " Highland Brand," 
which is represented as a sterilized article, is, on account of the 
sterilization, and because it contains no addition of foreign 
substances perhaps the only canned milk that, with proper care, 
could serve as exclusive food for young children. I mean to 
test its value in the near future. 

But in daily practice, in the cities, in towns or rural 
districts, there is, in my judgment, a means to overcome in a 
great measure all the difficulties cheaply. It is the only means 
which I can find which is simple, accessible to all classes, rich 
or poor. I mean the sterilization of cow's milk at home and 
the addition thereto (in case of very young or delicate cases) of 
substances such as will make it as near human milk as possible. 

In the first place, as to the preparation of the milk: 

It should be very fresh, and have been milked in a most 
cleanly manner in the cleanest of vessels, preventing any kind of 
dirt from contaminating any portion of it. Then to each half- 
pint or pint of milk distributed in nursing-bottles add about 
one-half to one tablespoonful of water previously boiled, and 
about two to four teaspoonfuls of chemically pure milk sugar. 
Then plug each bottle with a stopper of ordinary batting cot- 
ton and put in the sterilizer. 

A sterilizer consists of a steaming or boiling apparatus 
intended to raise the temperature of the milk sufficiently high 
to kill all adult germs and retard the growth of their spores, or 



332 THE BACTERIOLOGICAL WOULD. 

sufficiently high to destroy both the micro-organisms and their 
spores. The first method is termed pasteurization. 

The apparatus for sterilization consists of copper-bottom 
receptacles receiving a small amount of water which will readily 
generate steam in a few minutes when exposed to a sufficiently 
hot fire. The steam rises through the bottles full of milk. 
The Arnold sterilizer is a good example of this kind of 
apparatus. It can be purchased for about $3. 

An ordinary boiler or steam kettle, however, may be sub- 
stituted for these sterilizers. For instance, milk may be steri- 
ized by placing the milk bottles wrapped in cloths in such a 
vessel before the temperature is raised. 

Pasteurization means the raising of the temperature suffi- 
ciently high to destroy the adult germs and retard the growth 
of their spores into new organisms. It was found that a heat of 
180° F. to 200° F. thirty to forty minutes, followed with a rapid 
cooling of the milk, effects this aim without coagulating the 
casein in so hard and tough particles, as is done by higher tern, 
peratures. 

A temperature of 212° F. kept up twenty to thirty minutes 
effectually sterilizes the milk, so far, at least, as ferment microbes 
are concerned, and paralyzes other organisms, but the coagulation 
of the casein is naturally denser than at a lower heat. 

A temperature above 212° F. coagulates casein too seri- 
ously ; it becomes somewhat indigestible even to individuals 
secreting pancreatic juice in sufficient abundance, and, besides, 
all high temperatures seem to devitalize milk (as before stated, 
if this expression may be used) and make it less desirable for 
the nutrition of infants. Still, in summer time at least, it 
becomes necessary to raise the temperature to about 212° F., 
unless the milk can at once be placed in a cooler — a cool cellar, 
say, immediately after the operation. 

My practice has been usually to heat the milk prepared as 
above about thirty minutes at 212°F. immediately after milking 
in the morning in sufficient quantity to last all day and all 
night. Each bottle is plugged separately, and when opened for 
use, if there remains any amount of milk after nursing it is 
thrown away and new one opened at next nursing. 



THE BACTERIOLOGICAL WORLD. 333 

If it is found that the casein causes constipation and colic 
consequent thereon,, a little gelatine melted in boiled tepid 
water — say a teaspoonf ul of a saturated solution to each half- 
pint — is added to the milk before sterilization. This seems to 
act mechanically in englobing the casein when formed into 
particles of coagulum or curd. 

It will be seen by the foregoing that the objects of this 
method of preparing cow's milk are, first, to make it as near 
the constitution of human milk as we can in the present condi- 
tion of science ; second, to destroy or arrest all germs capable of 
producing any kind of fermentation or decomposition of the 
milk elements including the undigested casein. 

By this means we prevent the generation of irritant acids, 
ptomaines, and possibly toxalbumens and toxic proteids, which, 
we all know, many different micro-organisms are capable of pro- 
ducing. 

But it does more. If. per accident any germs of tuberculo- 
sis or other animal diseases are present in the milk they, too, will 
be paralyzed in their respective actions, and thus the milk will 
become purified. 



334 THE BACTERIOLOGICAL WORLD. 

SELECTIONS. 



CHEMICAL RESEARCHES ON MICROBIC SECRETIONS.- 
TRANSFORMATION AND ELIMINATION OF THE NITRO- 
GENIZED ORGANIC MATTER BY THE PYOCYANIC 
BACILLUS IN THE MIDST OF A DETER- 
MINED CULTURE. 



BY MESSRS. A. ARNAUD AND A. CHARRIN. 

Before the Academy of Sciences, Paris, France 

Is pyocyanine, this coloring matter so characteristic of the 
culture of the pyocyanic bacillus, in direct connection, as to 
quantity, with the functional activity of the bacillus? This is 
the first question which presents itself to the mind when we 
endeavor to give an account of the utilization and transforma- 
tions that the organic matter introduced as nourishment in a 
culture undergoes. 

The analysis shows that the formation of the pyocyanine is 
very secondary, its proportion varies within slight limits, from 
3 milligrams to 6 milligrams per litre, even in the cultures 
relatively colored sufficiently ; inversely, in cultures almost 
colorless, we can ascertain that a considerable quantity of 
nitrogenized organic matter has been transformed by the 
bacillus into products almost elementary. 

The principal products that are created are ammonia and 
carbonic acid. We have sought to follow the phases and the 
stages of these transformations and to study their mechanism. 

Our first trials had for starting point beef-broth with 
addition of nutritive salts ; but, in this medium so complex, it 
is difficult to follow the modifications of the organic matter; 
we had later recourse to a liquid artificial culture of determined 
chemical composition in order to restrain the field of researches. 

The following mixture is particularly favorable to the 

development of the bacillus : 

gr. 

PO 4 KH 2 0.100 

PO 4 Na 2 H+12 Aq . . .- 0.100 

CO 3 KH 0.134 

Ca CI 2 0.050 

Crystallized asparagine 5.000 



THE BACTERIOLOGICAL WORLD. 335 

Water, a sufficient quantity to make a litre. 

Twenty-four hours after inoculation of this liquid, the 
temperature being maintained between 77°F. and 86°F. the 
greenish blue color manifests itself clearly, and gradually 
becomes more pronounced until the sixth or seventh day. 

By exact dosing, we have followed, hourly so to speak, the 
progress of the assimilation of the asparagine. 

We thus notice that the proportion of ammoniacal nitrogen 
capable of being set free by MgO, small at first, increases unceas- 
ingly until the fifteenth or sixteenth day, when the culture 
stops for want of nourishment. 

The asparagine decreases very rapidly ; sixty hours after 
the beginning of the culture it is already impossible to reveal 
the presence of it ; on the other hand, at the outset we are 
able to ascertain the formation of aspartic acid* combined with 
the ammonia ; this acid is, however, assimilated immediately 
during its production ; it disappears about the seventy-second 
hour ; at this moment in fact, the examination of the curve 
of the diagram f indicates that almost all of the nitrogen 
is already passed into the state of ammonia, save, of course, 
that which has contributed to the formation of the protoplasm 
of the bacillus. It is necessary to take into consideration also a 
small proportion of nitrogen which enters into some combina- 
tions not yet definite, probably very interesting from a physio- 
logical point of view, and which we will notice to-day only to 
determine its relative quantity. 

RESPECTIVE QUANTITY OF THE SUBSTANCES WROUGHT AT THE EXPENSE 
OP THE NITROGENIZED MATTER. 

gr. 

Nitrogen contained in the 5 gr. of asparagine to 1 lit. 

of culture 0.933 

Nitrogen eliminated in the state of ammonia (A) by 

hydration of the asparagine under the influence per 100 

of the pyocyanic diastase 0.4655 50.0 



* The assimilation of the aspartic acid appears to be an essentially 
vital phenomenon, proper to the pyocyanic bacillus ; we have not been 
able to ascertain the presence of intermediary products between it and 
the ammonia. 

f The diagram was presented to the Academy for illustration. 



336 THE BACTERIOLOGICAL WORLD. 

Nitrogen eliminated in the state of ammonia by the 
vital action properly so-called of the bacillus on 
the aspartic acid, formed by the diastase 0.3835 41. 1 

Nitrogen found again in the protoplasm of the bacillus 

(the weight of which being 0.410 gr. ) (B) 0.0435 4.66 

Nitrogen in organic combinations not yet determined 

(D) 0.0385 4.04 

Nitrogen of the pyocyanine. Loss 0.0003 



Observations. — A. The volatile bases are constituted by the 
ammonia : We have analyzed the crystallized salts, oxalate and chloro- 
platinate ; there is formed, however, a trace of methylamine, the pres- 
ence of which we have been able to reveal by the appropriate treatment 
of a large quantity of oxalate of the total volatile bases. 

B. The weight of the microbes vary with the age of the culture ; 
it can attain 0.670 gr. towards the sixth or seventh day. 

C. In varying the weights of the asparagine ( we have employed 
20 gr. of this substance per litre), we obtain in the same time the same 
products and absolutely in jDroportional quantity. * * # 

D. The weights of these substances can be estimated at 0.300 gr., 
by the difference of the weights of the extracts of the filtered culture. 



* * * 



Let us add, then, that according, to the following experi- 
ments, the bacillus splits the asparagine by hydration by means 
of a diastase : in fact, if the liquid of a culture by the bougie 
has nearly no action on the asparagine in vitro, the bacilli col- 
lected on the bougie, washed and mixed in a solution of aspara- 
gine containing chloroform * in order to prevent all vital action, 
splits it in the following known equation CO. AzH 2 — OH. 
AzH 2 — CH 2 — C0 2 H + H 2 0=Co 2 AzH 4 — CH.AzH 2 — CH 2 — 
C0 2 H. We explain easily why this diastase remains fixed in 
the most part on the microbic protoplasm rather than passing 
in the filtered liquid, by the great experiments of Wurtzf on 
the fixation of the papaineon the fibrine. 



* This liquid remains acid, contrary to that which occurs in the 
culture where the microbe evolves. 

f Wurtz, Comptes rendus, t XC, p. 1379, t XCI, p. 787. 



THE BACTERIOLOGICAL WORLD. 337 

THE CONTAGIOUSNESS OF GONORRHCEA. 

In discussing the contagiousness of chronic urethral dis- 
charges, in the Journal of Cutaneous and Genito- Urinary 
Diseases, March, 1891, Dr. George Emerson Brewer, of New- 
York, says the practicing physician is frequently called upon to 
answer inquiries regarding the limit of the contagious stage of 
gonorrhceal urethritis. The question is a serious one, especially 
when considered in its relation to marriage, and should be 
regarded as equal in importance to that of syphilis. 

It has not infrequently been Dr. Brewer's experience to be 
consulted by young men, a few weeks or months before a con- 
templated marriage, with a history of one or more attacks of 
gonorrhoea in former years, and who believed themselves to be 
well ; yet who upon a careful examination presented the unmis- 
takable signs of a chronic urethritis. The only evidence of 
disease remaining in these cases frequently was the presence in 
the urine of small thread-like bodies, to which the name of 
Tripperfadden has been given by the German surgeons, who 
first described them and demonstrated their importance. These 
minute shreds are composed of mucus, pus and epithelium, and 
represent the secretions which adhere to any granular patch or 
area of chronic inflammation remaining on the urethral mucous 
membrane. 

In consulting the standard authors upon the subject of 
genito-urinary diseases, one is impressed by the marked differ- 
ence of opinion, regarding the contagiousness of chronic urethral 
discharges, held by those whose reputation and experience 
entitle them to the foremost position in the discussion of this 
question. As early as 1785, Kuhn called attention to the fact 
that the discharges resulting from a gonorrhoea remained con- 
tagious so long as they contained pus. The opposite opinion 
was held by Hunter, who denied the possibility of contagion 
from the gleety discharges of a chronic urethritis. This view 
was also shared by Bell and Ricord. In a paper entitled "Gon- 
orrhoea a Non-specific Disease," published in the New York 
Medical Independent in 1864, A. K. Gardiner strenuously denies 
the contagious element in any, save the discharges from the 
most acute stage of the disease ; and says regarding gleet that it 
is " allowably benign and innocuous." Noeggeratb, on the 



338 THE BACTERIOLOGICAL WORLD. 

other hand, in a paper published in 1872, concludes that a man 
who has once been the subject of a gonorrhceal urethritis never 
fully recovers ; that the disease invariably lingers in the glands 
and ducts emptying into the canal, and may at any time furnish 
a secretion which may infect those with whom he has sexual 
relations. He also states that nine-tenths of all women married 
to men who have had gonorrhoea sooner or later become the 
subject of incurable and painful inn 1 ammator}^ disease of ihe 
uterus, tubes or ovaries ; that this infection may take place 
rapidly, and manifest itself as an acute affection, or by means 
of a slow and unrecognized process to which he gives the name 
of "latent gonorrhoea." In a subsequent paper, read before 
the American Gynecological Association in 1876, the author 
reiterates these opinions, and adds that 90 per cent, of all cases 
of sterility can be directly traced to gonorrhoea. 

Without entering into any discussion regarding the cor- 
rectness of these views, which have been the object of consider- 
able criticism, the fact remains that these papers had the effect 
of calling attention to a source of female disease and suffering, 
the importance of which had not, until their publication, been 
adequately recognized. This difference of opinion upon so 
important a question is not to be wondered at, when we con- 
sider that, at the time these views were enunciated, nothing 
definite was known regarding the etiology of this disease. 
Since the discovery by Neisser, in 1878, of the gonococcus, and 
the (Dr. Brewer thinks) establishment of its relationship to 
this disease, but one opiniou can logically be held by those who 
accept his theory of gonorrhceal inflammations, and that is, 
that all secretions containing this micro organism are capable 
of transmitting the disease under favorable conditions. In his 
recently published work upon this subject, Ernest Finger 
emphasizes this point, and states regarding marriage, that it 
should be absolutely prohibited in all cases where the existence 
of a chronic urethritis is evidenced by the presence of the 
"morning drop" or Tripp erf adden in the urine, until the fol- 
lowing facts have been established : 

First. That after from two to four weeks of daily observa- 
tion, the secretions from the urethra are found to he free from 
pus and made up ivholly of epithelial cells. 



THE BACTERIOLOGICAL WORLD. 339 

Second. That no gonococci can be detected by the micro- 
scope, even after a purulent discharge has been established by 
the employment of irritative injections of corrosive sublimate 
or nitrate of silver ; and 

Third. That neither prostatitis nor stricture exists. 

Dr. Brewer says that, upon first becoming acquainted with 
the views expressed by Finger, he regarded his conditions as 
unnecessarily severe, and his opinion of the danger as greatly 
exaggerated. But a somewhat remarkable case, which came 
under his observation, illustrated in a most striking manner the 
fact, that, unless some of the precautions advised by Finger had 
been insisted upon, the responsibility of a dangerous illness 
would justly have rested upon his head. The case is reported 
by Dr. Brewer, and fully justifies the inferences he draws from 
it. — Medical and Surgical Reporter. 



LECTURE ON PHAGOCYTOSIS AND IMMUNITY 



BY DR. ELIAS METSCHNIKOFF. 

Pasteur's Institute, France. 
[concluded.] 

Now, in determining the intervention or non-intervention 
of the leucocytes in this war between the organism and the bac- 
teria, a very great part is played by the sensitiveness of these 
cells to external influences, and especially to the chemical com- 
position of their environment. The leucocytes are powerfully 
attracted by many micro-organisms and the resultants of their 
growth, and as powerfully repelled by others and their result- 
ants, or, as it is expressed, they have a positive chemiotaxis for 
certain microbes, a negative chemiotaxis for others. The exist- 
ence of these chemiotactic properties has been so clearly 
proved of late by the researches of Leber, Massart and Bordet, 
and Gabritschewski, that I need not enter into a fuller expla- 
nation of the subject here. Where negative chemiotaxis mani- 
fests itself, there, being shunned by the white corpuscles, the 
parasites freely propagate themselves and induce the death of 
their host. Nevertheless, this chemiotaxis is not immutable, 



340 THE BACTERIOLOGICAL WORLD. 

and the cells can become accustomed to substances from which 
they shrank at first — a negative may thus be transformed into a 
positive chemiotactic state. Such obtains in acquired immun- 
ity; the cells which, in the unvaccinated animal never included 

the bacteria, now, in the vaccinated, take them up readily. 
* * * * * * * * 

There is not a single portion of the theory which I have 
just expounded but has encountered a lively opposition. Even 
the fundamental fact that the phagocytes are capable of includ- 
ing the microbes has had doubts thrown upon it ; it has been 
held that the latter insinuate themselves into the former. 
Only after successive series of observations upon the phago- 
cytes and the living microbes, has it been proved that assuredly 
it is the phagocytes which, by the aid of their pseudopodia, 
themselves include the micro-organisms. The observer can see 
the whole process in the case of immobile bacilli, can see the 
leucocyte approach, send out pseudopodia, and gradually 
include the individual bacillus. Or, conversely, in cases of 
negative chemiotaxis, one can, in blood taken from the monkey 
during the access of relapsing fever, observe the actively mov- 
ing spirilla come into contact with a leucocyte, and even 
become attached by one end to its surface ; yet, however active 
the movement, one never finds that the spirillum succeeds in 
piercing the surface and gaining an entrance. If it be sug- 
gested that this entry may take place in consequence of the 
force of active growth and elongation of bacilli, then, apart 
from the fact that here but one set of cases is embraced, it can 
be determined that this force is too feeble — it can be seen that, 
during the active growth of the anthrax organism in the blood, 
the elongating chains of bacilli curve in and out between the 
corpuscles, but never penetrate the cells. 

From another side, the objection has been formulated that, 
in many cases, the organism gets rid of its invaders without the 
aid of the phagocytes. According to^ those who support this 
objection, this happens in the anthrax of pigeons (Czaplewski ), 
and of refractory rats (Behring, Franck ), in symptomatic 
anthrax of various refractory animals ( Rogowicz ), and in the 
septicaemia of vaccinated guinea-pigs* due to the vibrio Metsch. 
nikovi ( R. Pfeiffer ). A re-examination of the cases here 



THE BACTERIOLOGICAL WORLD. 



341 



adduced has, however, shown that in each a very considerable 
phagocytosis can be proved, and that the negative results of 
the above observers have been due to insufficient methods of 
observation. 




- c/ 



Fig. 3.— Anthrax of pigeon (an animal but slightly susceptible to the disease;, to 
show stages of destruction of bacilli by phagocytes. 1 and 2, macrophages ; 
1, from exudation from eye of refractory bird ; 2, from muscle of region of 
inoculation of bird that succumbed ; 3, 4, 5. microphages — all from eye 
twenty-seven hours after inoculation ; a, a, unaltered bacilli ; 61, 52, 63, bacilli 
becoming more and more degenerated and indistinct ; c c, debris of bacilli 
( Zeiss 1-18, ocular 3 ) . 

While accepting that the phagocytes do truly absorb the 
micro-organisms, other opponents of the theory have urged 
that these cells are only capable of including micro-organisms 
already killed by other means, and that living microbes are 
solely to be found within the cells in those cases where there 
has been a fatal ending — in tuberculosis, mouse septicaemia, 
and so on. Against this may be brought the fact determined 
by Lubarsch, that the phagocytes of several animals, refractory 
to anthrax, take up living bacilli that have been injected with 
greater eagerness than they include those which have been 
killed before injection. But, further, this objection may be- 
disposed of by direct observation of bacteria undergoing devel- 
opment from within the interior of phagocytes after the latter 
have been destroyed by a substance which is at the same time 
a favorable medium for bacterial growth, as, for instance, beef 
broth. Such observations have been made upon pigeons, ren- 
dered immune to anthrax. 



342 THE BACTERIOLOGICAL WORLD. 

During the last year or two, great stress has been laid upon 
the fact that the bodily humors themselves possess most 
marked bactericidal properties, and, in fact, against the 
theory of phagocytosis has been brought another, based upon 
this power of the humors to destroy the microorganisms. 
Observer after observer has remarked, that, in blood plasma, 
defibrinated blood, blood serum, and in the blood as a whole, 
in the removed aqueous humor and other fluids and exuda- 
tions [of the body, many species of bacteria perish, after a 
longer or shorter interval ; and, forthwith, an endeavor has 
been made to find in these facts some elucidation of the 
phenomena of immunity. Yet, the more deeply one examines 
into the question, the more one is convinced that no relation- 
ship exists between the two. Thus it happens often, that the 
bactericidal property is more developed in susceptible species 
than in refractory ; so with regard to the anthrax bacilli, in 
the very sensitive rabbit the bactericidal properties of the 
humors are more pronounced than they are in the refractory 
dog ; and Beliring and Nissen, the two who almost simul- 
taneously first drew our attention to these phenomena, in their 
combined research, recently published, admit that, as against 
the bacteria of anthrax, pneumonia and diphtheria, this 
bactericidal property exists to the same degree in the juices 
of animals of the same species, whether they be susceptible or 
have been rendered immune. Often, again, it has been deter- 
mined that the blood removed from the organism has a 
greater power of destroying bacteria than it has within the 
organism. A small quantity of blood withdrawn from the 
body will, in certain instances, kill a mass of bacilli greater 
than that which, injected into the circulation, would inevitably 
cause death. Evidently, therefore, in this bactericidal influ- 
ence, extravascular phenomena enact an important role — phe- 
nomena, that is, which have no connection with what occurs in 
the living refractory organism. 

From another point of view, strong arguments have been 
directed against this theory of the tissue fluids. It has been 
shown, specially by the researches of M. Haffkine, that the 
death of the bacteria transported into organic fluids is 
largely due to the sudden change of medium, and that, in 



THE BACTERIOLOGICAL WORLD. 343 

passing from one medium to another by successive slight modi- 
fications in the fluid of growth, it is easy to make bacteria live in 
fluids which, when the change of environment has been abrupt, 
swiftly lead to their destruction. 

In order to gain an idea as to the part played in the refrac- 
tory animal, by the fluids and the phagocytes respectively, the 
endeavor has been made to separate the two by placing under 
the skin of frogs ( which are naturally immune to anthrax) 
minute packets formed of filter paper, or of animal membrane, 
and containing the bacilli. The paper, while permitting the 
passage of fluid, wards off the wandering amoeboid cells for a 
certain time. Shielded in this way from the phagocytes, 
though exposed to the action of the juices, the bacilli grow 
well, and produce the characteristic felted mass of anthrax fila- 
ments. Baumgarten has not been able to confirm this experi- 
ment, but Hueppe and Lubarsch have repeatedly verified it. 

But it is not even necessary to take these precautions, in 
order to assure one's self that anthrax spores germinate in the 
juices of refractory animals. Recently, for instance, M. 
Trapeznikoff has found that, when these spores are injected 
into the dorsal lymph sac of the frog, they constantly tend to 
develop into bacilli, whose further growth is stopped by the 
phagocytes, which include them, along with such spores as 
have not had time to germinate. Eventually the bacilli so 
absorbed are digested by their hosts, while the included spores 
remain intact, although incapable of giving birth to bacilli for 
so long a time as the phagocytes remain alive. And I might 
adduce other similar cases. Such a comparative examination 
proves that in the living body the bactericidal property resides 
in the phagocytes, and not in the fluids. 

Still, it may be urged that possibly these cells, which can 
thus devour and destroy the living microbes, are only in a posi- 
tion to attack bacteria whose virulence has already been lessened 
by other means. Were this so, the microbes present in a 
refractory organism should behave, not like parasites, but as 
simple, inoffensive saprophytes. Hence, these microbes — 
powerless to produce upon a refractory soil the toxic substances 
which render them pathogenic and dangerous — should easily 



344 THE BACTERIOLOGICAL WORLD. 

be included and destroyed ; so that, according to this hypoth- 
esis, which has frequently been brought forward, the phagocytes 
play a purely secondary and dependent part, waiting until the 
microbes are weakened before they seize upon them. In. favor 
of this view, the fact has been cited that certain micro- 
organisms cultivated in the blood, or serum, of vaccinated 
animals become attenuated, so that they no longer induce a 
fatal disease. The bacillus anthracis grown in the blood of 
vaccinated sheep no longer kills rabbits, and, according to 
Koger, the streptococcus erysipelatosus grown in the blood of 
vaccinated rabbits only occasions a slight and passing disturb- 
ance in susceptible members of the same species. But, here 
again, we are dealing with fluids withdrawn from the body, 
and so modified in various ways. Let us make an observation 
more strictly to the point : Take, for instance, a rabbit vacci- 
nated against anthrax and inoculate it with anthrax bacilli, 
thus allowing these to exist directly within the refractory 
organism. Such bacilli as are not destroyed preserve their 
virulence for a sufficiently long period, and it is possible to kill 
a guinea-pig with a drop of exudation taken from the region of 
injection thirty hours after subcutaneous inoculation, eight 
days after inoculation into the anterior chamber of the eye. A 
sojourn of so long duration within the vaccinated organism, 
then, has not deprived the microbes of their virulence, although 
twenty -four hours suffices to completely attenuate the bacilli 
cultivated in the removed blood of vaccinated sheep. 

Years ago, it was established in M. Pasteur's laboratory 
that the refractory organism, instead ot being an unfavorable 
soil for the preservation of virulence, tends the rather to rein- 
force this property. To exalt the virulence of an attenuated 
micro-organism, one always employs, not animals very suscep- 
tible to the specific disease, but those which are slightly suscep- 
tible, or to it may be, under many circumstances, refractory. In 
this manner the most active anthrax virus has usually been 
obtained by passage through birds, notably fowls ; the greatest 
virulence of chicken cholera was gained by passage through the 
vaccinated cock, and quite recently M. Malm has shown that 
passage of the anthrax bacillus through the organisms of dogs, 
which of*[all mammals are the most refractory in this respect^ 



THE BACTERIOLOGICAL WORLD. 345 

increases its virulence in a most remarkable manner, so that the 
general law may be laid down that an organism which i3 but 
slightly susceptible or is refractory is able not only to preserve, 
but even to exalt, the virulence of bacteria. The principal 
argument in favor of the hypothesis, that pathogenic micro- 
organisms become simple inoffensive saphrophytes when they 
find themselves in a refractory region, loses therefore its raison 
d'etre. 

M. Bouchard, in his objection to the theory of phagocytosis, 
may be regarded as introducing but a modification of this 
hypothesis. He holds that pathogenic bacteria placed unde r 
favorable conditions give rise to substances which hinder the 
inflammatory process, and that only when these inhibitory sub- 
stances are inadequately represented do the cells intervene. 
When, therefore, the organism rendered refractory by vaccina- 
tion becomes an unfavorable soil for the production of these 
inhibitory bodies, the bacteria can no longer prevent the inflam- 
matory reaction ; free emigration of the leucocytes ensues, these 
cells seize upon the impotent microbes, and put a stop to their 
further growth. In this theory, the part played by the phago- 
cytes is again secondary, depending upon a dearth of anti-inflam- 
matory substance. 

If the theory could be accepted in certain cases, it is never- 
theless inapplicable as a general rule. In all those affections 
which are characterized by the absence of leucocytes upon the 
field of battle there is certainly no lack of inflammation. The 
very reverse obtains. In anthrax affecting small mammals, 
just as in the vibrionic septicaemia of pigeons and guinea-pigs 
and other analogous diseases, we find that there is a very distinct 
dilatation of the vessels, accompanied by great exudation ; the 
inflammatory reaction is well marked, nothing is wanting save 
the determination of the white corpuscles. Or employing yet 
further that affection, which is, as it were, the touchstone of 
the bacteriologist, a still clearer proof of our contention is to be 
gained, if we inoculate a rabbit on the one ear with a small 
quantity of virulent, on the other with a like quantity of atten- 
uated, anthrax virus. In the course of a few hours the external 
signs of inflammation are far more conspicuous in the former ; 
the vessels are greatly enlarged, and there is literally a huge 



346 THE BACTERIOLOGICAL WORLD. 

exudation of clear serous fluid into the part ; in the latter the 
external signs are less prominent, but examination of the seat 
of inoculation shows it to be packed with leucocytes. Conse- 
quently, the phenomenon we are discussing is to be explained 
not by an absence of the inflammatory process, but much more 
satisfactorily by a negative chemiotaxis of the leucocytes, which, 
instead of being attracted by the bacterial products, are repelled; 
where the animal is vaccinated or refractory a much slighter 
inflammation is sufficient to produce an abundant emigration of 
the leucocytes. 

During the last few days Behring has brought forward 
another view which would explain immunity in a wholly differ- 
ent way. According to him the bacteria can live and even pre- 
serve their virulence in the refractory organism, but the loxines 
excreted by them now undergo a modification so as to be ren- 
dered completely inoffensive for the animal. And to this "tox- 
icide property" of the organism is to be attributed the essential 
quality of the immune state. It is impossible to pronounce 
upon the arguments that have led up to this theory, for as yet 
they have not been circumstantially set forth, but already one 
can declare that such a theory is in no wise applicable to the 
phenomena of immunity in general. In three diseases remark- 
able for their pronounced toxic character — vibrionic septicaemia, 
pyocyanic disease, and hog cholera affecting the rabbit — as 
shown by the experiments of Oharrin, Gamaleia and Selander 
the toxines are so little attacked by the refractory organism that 
the same quantity of these substances (freed from bacteria) suf- 
fices to kill an animal very susceptible to one or other disease, 
and an animal vaccinated against it and thus completely 
immune. So, too, non-fatal doses of these toxines produce in 
animals of the two categories the same febrile and inflammatory 
reactions. The proof is clear that there is no special destruc- 
tion of toxines in the refractory animal, and that the " toxicide 
property," if it exists, is not one whit more developed after 
vaccination than before. Passing in review all these counter 
theories we see that each of them can only be applied to a certain 
number of facts ; in some an attenuating or even bactericidal 
influence of the juices is relied upon, in others an anti-inflam- 
matory action ; in yet others a toxicide property. Still the 



THE BACTERIOLOGICAL WORLD. 347 

phagocytic reaction is the only constant in all those cases of 
immunity and recovery that have as yet been sufficiently 
studied, and while certain of the factors mentioned (the atten- 
uating and toxicide properties) do not in the least touch upon 
the continued existence or otherwise of the micro-organism, the 
bactericidal power of the phagocyte puts an end to the parasite 
itself, and thus at a given moment prevents further manifesta- 
tion of its virulence, or preserves the animal attacked at a time 
when the toxicide properties would be found wanting, and the 
microbe remaining alive would consequently gain the upper 
hand. 

But while thus placing before you the important part played 
by the phagocytes, I do not wish it to bethought that these cells 
are unaided in their contest by other defensive means possessed 
by the organism. This is far from being my view. Thus, in 
the febrile reaction, we see a puissant auxiliary very definitely 
favoring the work of the phagocytes. This febrile reaction has 
only to be inhibited — as was done by M. Pasteur in the anthrax 
of fowls — and animals naturally refractory to the affection suc- 
cumb to the ravages of the bacilli. It is not possible at the 
present time to state fully and accurately all these influences 
which are associated in aiding phagocytic action, but already, 
we have the right to maintain that, in the property of its amoeboid 
cells to include and to destroy micro-organisms, the animal tody 
possesses a formidable means of resistance and defense against 
these infectious agents. — The British Medical Journal. 

[the end.] 



THE NUMBER AND APPEARANCE OF TUBERCLE BACILLI 
IN THE SPUTUM AFTER TUBERCULIN INJECTIONS. 

BY L. BREMER, M. D. 

Before the St. Louis Medical Society. 
I have brought with me to-night a specimen of tuberculous 
sputum for the inspection of the Society. The specimen has 
been prepared after Ziche's method, with carbolo-fuchsine» 
The sputum was handed to me for examination by Dr. Mul- 
hall. It is obtained from a patient who is undergoing the 
Koch treatment. 



348 THE BACTERIOLOGICAL WORLD. 

Among the first published reports on the Koch method 
was one of Fraentzel, of Berlin, who claimed that the form of 
tubercle bacillus was materially changed by the tuberculin 
when introduced into the circulation. He asserted that after 
injection the bacilli lost much of the staining facilities ; that 
they became smaller and more slender, assumed the biscuit 
shape or that the rods were composed of a number of coccus- 
like formations. 

If the sputum which I submit to your examination is an 
example of what Fraentzel saw and described, I must say that 
I have seen, time and again, like others, the same in the many 
sputum examinations which I have made since the day that 
Ehrlich's method was first published. 

Some time ago, I received from a friend, one of the slides 
which were advertised and for sale in Berlin, showing the bacilli 
as they appear after tuberculin injections. The forms that I 
saw there were old acquaintances. 

In the specimen which I have to-night placed under the 
microscope, some of the microbes show the forms which have 
been described as characteristic of tuberculin action. 

They are present in immense numbers, so that the droplet 
of sputum, which was subjected to the staining process, may 
justly be said to have been a pure culture of tubercle bacilli, 
exhibiting various stages of a retrogressive metamorphosis. 

To my mind, the explanation of this often observed fact is 
the following, viz. : 

Nearly all observers argue that cough and expectoration 
increase after tuberculin injections. With increased cough, 
little round, whitish, more or less globular bodies, which are 
often formed in cancers, and which consist of almost pure cul- 
tures of the parasite, are expelled and are made accessible to 
microscopical examination. Iodide of potassium, which has of 
late been proposed instead of tuberculin by an investigator, 
for diagnostic purposes, will probably have the same effect. 

Specimens like the one which I have prepared for your 
examination, I have often seen in ante tuberculin days. — St. 
Louis Medical and Surgical Journal. 



THE BACTERIOLOGICAL WORLD. 349 

SCIENTIFIC GLEANINGS. 



AMMONIACAL FERMENTATION OF URINE. 



BY VERANUS A. MOORE, M. D. 

[ CONCLUDED. J 

DESCRIPTION OE NEW GERMS. 

In the light that we already have concerning this class of 
organisms and in the present unsettled condition of bacterio- 
logical classification,^ seems more desirable to omit the assigning 
of specific names to the micrococci about to be described, but 
simply differentiate them with reference to their morphological 
characters and their growth in certain culture media. They 
will, therefore, be designated by the first letters of the alphabet, 
printed in small capitals, as micrococcus a. mic. b, etc. In 
comparing the morphological and cultural characters of these 
germs with the description of the micrococci described by Pas- 
teur, Fluegge and Miquel I have failed to find sufficient resem- 
blances to establish the identity of any of these micrococci with 
those heretofore described. Their effect upon urea in nutrient 
solutions is, however, practically the same. 

Micrococcus a. — A micrococcus varying in diameter from 
0. 6 pi to 1. 2 /j. ; average, 0. 9 jx. When grown in bouillon the 
cocci appear most usually in small clumps and in the form of 
diplococci. There seems to be a marked disposition in the 
cocci to be closely united in pairs. In acid bouillon they are 
frequently observed united in short chains. 

Agar {surf ace growth ) at 96. 8° F. — After 24 hours round, 
convex, grayish colonies appear with a glistening surface and 
sharply defined border. When they are separated by a distance 
of one-half inch or more they are about 1 millimetre in diam- 
eter. After growing for several days the colonies become 
opaque, whitish, but otherwise unchanged in appearance. The 
colonies are quite brittle. The growth in the condensation 
water is quite vigorous, giving it a clouded appearance. 

Gelatine ( needle culture ). — In nutrient gelatine the growth 
is quite vigorous at 70° F. Along the needle track, after 24 



350 THE BACTERIOLOGICAL WORLD. 

hours, a grayish, opaque, somewhat granular line appears, which 
consists of crowded colonies. At the surface the growth is 
more vigorous, forming an elevated glistening band around the 
needle puncture. This may be due to the greater number of 
germs collected there. On the second or third day the gelatine 
begins to liquefy along the entire length of the needle track. 
At first the liquefaction is generally more rapid on the surface, 
giving a cone-like appearance to the liquid portion, the base 
being at the surface ( PL I, Fig.l). Very of ten the liquefaction 
is in the form of a cylinder. The entire mass of a gelatine is 
liquefied in from 8 to 20 days. In a higher temperature (75° 
to 80° F. ) the growth is more rapid. The liquid gelatine holds 
in suspension fine granular masses of germs 1-10 to 1-4 millimetre 
in diameter. A considerable quantity of a grayish sediment 
collects in the bottom of the tube ; also a fine granular pellicle 
forms over the surface. Both the pellicle and sediment are 
very viscid. The liquefied gelatine will not set, showing that 
it has become peptonized. 

In alkaline bouillon the growth is. very feeble, the liquid 
becoming barely clouded, even after several days' standing. In 
acid bouillon the growth is at first also feeble, but on the second 
day it is very much increased, the liquid is quite clouded, and 
a granular pellicle forms over the surface. The acid reaction is 
not appreciably changed In sterile milk at 96. 8° F. this germ 
coagulates the casein after about two weeks standing. It grows 
very vigorously. The reaction remains slightly acid. A slight 
sour-milk odor is emitted. 

Sterile urine at 96. 8° F. — This germ multiplies very rap- 
idly in this liquid. After 18 hours it becomes quite clouded 
with the formation of a grayish sediment. Its acid reaction is 
changed to a feebly alkaline one, which becomes very strong 
after 36 to 48 hours. The odor is penetrating, resembling that 
emitted by ammonia water. In acid bouillon plus urea at 
96. 8° F. this germ grows very rapidly. After 24 hours the liquid 
is very clouded, with considerable sediment formed in the 
bottom of the tube. The reaction is changed to a strong alka- 
line one. A slight ammoniacal odor is emitted. In the acid gel- 
atine plus urine the growth is more vigorous and the liquefaction 



THE BACTERIOLOGICAL WORLD. 351 

more rapid than in the ordinary gelatine. With the liquefac- 
tion its reaction is changed to a strong alkaline one. The 
liquid gelatine becomes perfectly cleared, with a somewhat 
viscid sediment. 

This germ is easily stained with the aniline dyes. It does 
not take the Gram stain. 

Micrococcus b. — A micrococcus varying from 1.0 // to 1.8 
}x in diameter ; average, 1.4 jx. It occurs single, in pairs., and 
small clumps. It is occasionally seen in short chains, especially 
when grown on agar or gelatine. 

Agar {surface growth} at 96. 8° F. — The growth of this 
germ on agar resembles in its general appearance that of the 
micrococcus A. It differs from that germ in two respects : (1) 
The growth on the surface is very viscid. (2) The condensation 
water remains clear, with a grayish, somewhat viscid growth 
at the bottom. 

Gelatine {needle culture) at 75° F. — After 24 hours a 
grayish granular line appears along the needle track. At the 
surface the growth is more vigorous, spreading and liquefying 
the gelatine in a thin layer over the entire surface. The growth 
along the needle track does not increase, but the gelatine grad- 
ually liquefies from the surface downward until from one-half 
to two-thirds of the gelatine has become liquid. The liquefac- 
tion does not extend beyond that point ( PI. I, Fig. 2 ). A 
more or less thick, whitish membrane forms over the central 
portion of the surface, rarely forming a complete pellicle. A 
layer of grayish, viscid growth forms over the surface of the 
non-liquefied gelatine. The liquid portion holds in suspension 
grayish granules, which are occasionally so fine as to give a 
uniform clouded appearance to the liquid gelatine. The lique- 
faction is completed in from one to two weeks. 

In bouillon at 96. 8° F. the growth is very feeble. After 
24 hours the liquid is very faintly clouded. In from two to 
four days a somewhat stringy growth appears in the upper part 
of the liquid ; also granular masses, varying from 1 to 2 milli- 
metres in diameter. These are quite viscid. In acid bouillon, 
after several days a few granular masses are formed on the 
surface ; also a whitish deposit on the sides of the tube at the 
surface of the liquid. 



352 THE BACTERIOLOGICAL WORLD. 

In sterile milk this germ, after about two weeks, coagulates 
the casein, leaving a clear supernatant liquid. 

Sterile urine at 96. 80 F. — This germ grows rather freely in 
urine. After 24 hours the liquid is very faintly clouded and 
neutral in reaction. After 48 hours it is strongly alkaline and 
clouded with the formation of a small quantity of sediment. 
In acid bouillon plus urea the growth is not vigorous. After 
24 hours the reaction is feebly alkaline. At the end of two to 
three days the reaction is strongly alkaline and the liquid quite 
cloudy — a slight penetrating odor. In acid gelatine containing 
urea the growth is more vigorous than in the ordinary gelatine. 
The liquefied gelatine is strongly alkaline in reaction. 

This germ is easily stained with the aniline dyes. It takes 
the Gram stain. 

Micrococcus c. — A micrococcus varying in diameter from 
8 fj. to 1.5 jd ; average, 1.2 yw. It occurs single, in short 
chains, and small clumps. There are also diplococci forms. 

Agar (surface growth) at 96.8° F. — On this medium the 
growth does not differ appreciably from that of micrococcus a. 

Gelatine.— This germ liquefies gelatine very slightly, so 
that the growth, which at first is upon the surface, after several 
days becomes slightly depressed, never more than one to two 
millimetres. The growth along the needle puncture is whitish, 
opaque, viscid, and quite thick. The colonies along the needle 
track are very small. They do not liquefy the gelatine (PI. 
I, Fig. 3). 

In bouillon the growth is very feeble ; after 24 hours the 
liquid becomes faintly and uniformly clouded. There is no 
further development. Sterile milk inoculated with this germ 
and kept at a temperature of 96.8° F, becomes thickened in the 
lower half of the tube after about six days. There is no 
separation of the casein from the aqueous portion ; no odor ; 
slight acid reaction. 

Sterile urine. — In this liquid the growth is feeble. On the 
day following its inoculation the reaction is faintly alkaline, 
the liquid barely clouded. On the third day there is a thin 
granular deposit on the sides of the tube, and the reaction is 
strongly alkaline, ammoniacal odor. In acid bouillon plus urea 



THE BACTERIOLOGICAL WORLD. 353 

at 96.8° F. the liquid becomes clouded on the day following the 
inoculation, with the formation of a considerable quantity of 
sediment ; strong alkaline reaction, penetrating odor. After 
ten days' standing it becomes clear, with a sediment. It 
remains strongly alkaline. In acid gelatine plus urea the 
growth is very feeble ; no liquefaction. 

It is easily stained by the ordinary methods. When stained 
according to Gram a great majority of the cocci take the brown 
stain perfectly, A few ( about one in fifteen ) retain the blue 
stain. These are of the maximum size (1.5 pi). 

Micrococcus d. — A micrococcus varying in diameter 
from 0.7 yw to 1.2 m; average, 0.9 jj.. It grows mostly usually 
in small clumps and in pairs. It is very rarely seen single. 

Agar (surface growth) at 96.8° F. — When not crowded the 
isolated colonies, after 24 hours, are from 1 to 1 1-2 millimetres 
in diameter, round, flat, with a smooth border and sharply 
defined margin ; a considerable growth in the condensation 
water. The colonies at first are grayish white, glistening, but 
after several days they become a light cadmium yellow. The 
size is very slightly increased. When grown on agar containing 
5 per cent, glycerine the growth is more feeble, but the color 
more intense. 

Gelatine (needle cultures). — Along the needle track the 
colonies develop after 24 hours as minute grayish points. After 
several days they attain to a diameter of 1-8 to 1-4 millimetre, 
spherical, granular, yellowish. On the surface the growth is 
spreading, glistening, of a deep cadmium yellow. The growth 
about the needle puncture appears in more or less concentric 
bands. The outer bands also have a radiating appearance, 
produced by alternate lighter and darker lines extending out- 
ward from the more central portion. After about two weeks 
the growth becomes slightly depressed below the surface ( PL 
I, Fig. 4 ). 

In bouillon the growth is feeble. After 48 hours, 96.8° F. 
the liquid becomes clouded in the upper portion of the tube, 
with a small quantity of a grayish deposit formed on the side of 
the tube. It produces no change in the appearance of milk. 

Sterile urine. — In this liquid the growth is abundant ; the 
liquid is heavily clouded after 24 hours ; the reactions strongly 



354 THE BACTERIOLOGICAL WORLD. 

alkaline. After from two to three days a considerable quantity 
of sediment is formed in the bottom of the tube. This germ is 
the most rapid in its transformation of acid into alkaline urine 
of any of the germs here described. Under its influence acid 
urine will become decidedly alkaline in eighteen hours. In acid 
bouillon containing urea the growth is vigorous. In 24 hours 
the liquid is very cloudy and strongly alkaline in reaction. In 
acid gelatine plus urea the growth is more feeble than on the 
ordinary gelatine. It takes the Gram stain. 

Micrococcus e, — A micrococcus varying from 0.9 yw to 
1.5 }jl in diameter ; average, 1.2 yu. It occurs most usually in 
small clumps and in pairs. It is frequently found single, often 
in short chains. 

Agar {surface groivth) at 96.8° F. — After 25 hours quite 
fleshy, round, convex colonies appear, 1 millimetre in diameter 
when not crowded. They are of a grayish yellow color, opaque. 
After several days the growth becomes a much deeper yellow. 
The condensation water is clear, with a viscid, flaky growth at 
the bottom. The surface growth is not viscid. 

Gelatine. — The growth in this medium resembles that of 
the micrococcus b. It differs from it in the color of the mem- 
brane and sediment, which are cadmium yellow ( PI. I, 
Fig. 5). 

In bouillon the growth is feeble. It does not produce any 
apparent change in the appearance of milk. 

Sterile urine. — The growth in this liquid is very slow. On 
the second day after inoculation it becomes feebly alkaline, with 
the formation of a granular deposit on the sides of the tube. 
In acid bouillon containing urea the growth is more vigorous. 
After one day, at 96.8° F. the reaction becomes strongly alkaline, 
emitting a penetrating odor. It liquefies acid gelatine contain- 
ing urea very slowly. The liquefaction does not extend more 
than three to five millimetres below the surface. The liquid 
gelatine is strongly alkaline in reaction. This germ is readily 
stained by the ordinary methods. It takes the Gram stain. 

The characteristic properties of these micrococci, especially 
their reaction on nutrient gelatine, are more or less modified by 
cultivation. Their power to break up urea, however, does not 
seem to be diminished. 



THE BACTERIOLOGICAL WORLD. 355 

I have examined, after the methods previously described, 
twelve specimens of urine. Of these, seven contained onlv the 
micrococcus a, two the micrococcus c, one the micrococci a and 
B, one the micrococci B and d, and one the micrococci A, c 
and e. Eight of the specimens were taken from^the same indi- 
vidual, six of which contained (of the germs described) the 
micrococcus A only, and two the micrococcus c. The other 
four specimens were obtained from four other individuals, one 
of which contained only the micrococcus A. 

In addition to the above, one specimen of urine from a per- 
son suffering with cystitis was examined. The urine was 
clouded, strongly alkaline in reaction, and contained a consid- 
erable quantity of crystals, principally triple phosphates, pus 
cells and amorphous salts. A large number of micrococci were 
seen in a cover-glass preparation. The bacteriological exami- 
nation showed that a large number of these were the micrococ- 
cus A. Thus far this is the only specimen of urine from an 
authentic case of cystitis that I have been able to obtain in a 
condition suitable for a bacteriological examination. 

I have also isolated several germs from external sources that 
are equally as active in the production of ammoniacal fermenta- 
tion as those already described. Two of these, a bacillus and 
micrococcus, were obtained from decomposed urine that was 
previously sterilized and then exposed to the atmosphere of the 
laboratory by removing the cotton wool stoppers from the tubes 
•containing it. The sterile, acid urine became clouded and alka- 
line in from three to five days. A second bacillus was obtained 
by inoculating a tube of sterile urine with several drops (about 
1 cc.) of Potomac water and allowing it to stand in an incuba- 
tor, where it became clouded and alkaline in twenty-four hours. 
The bacillus was isolated from the other germs by means of 
plate cultures. A micrococcus was also found in saliva which 
is arranged in fours (tetracoccus), that is very active in trans- 
forming urea into carbonate of ammonia. These germs have, 
as yet, not been carefully studied, and their isolation is men- 
tioned here only to illustrate the general distribution of bacteria 
possessing this property, as well as to confirm similar statements 
made by other workers in this line. 



356 THE BACTERIOLOGICAL WORLD. 

SOLUBLE FERMENTS. 

In the last decade Musculus (9) announced the discovery 
of a soluble ferment which he obtained from ammoniacal urine 
that would, when introduced into fresh urine, transform its 
urea into carbonate of ammonia. The name urase was proposed 
for this substance, as it was supposed to belong to the same 
class of ferments as ptyalin diastase, etc. "The method of iso- 
lating this ferment was reported by Musculus to be one of sim- 
ple precipitation. The results, however, that have been reported 
by others who have attempted to isolate this substance, have 
been, so far as I can learn, negative in character. P. Miquel, 
after a long series of experiments, in many of which the method 
of Musculus was carefully carried out, concludes with Prof. 
Leube, that the existence of a soluble ferment in urine is not 
sufficiently proven to be accepted without further demonstration. 
Thus far I have made no experiments for the purpose of isolating 
a soluble ferment. Whether there is such a ferment or not will, 
together with many other interesting questions that these pre- 
liminary investigations have suggested, have to remain unan- 
swered until other and more extended investigations shall be 
made. 



THE BACTERIOLOGICAL WORLD. 



357 



Plate I, 







i i 





Fig. 1. 



Fig. 2. 



Fig. 3. 



Fig. 4. 








k 





mm M 




Fig. 5. 



358 THE BACTERIOLOGICAL WORLD. 



DESCRIPTION OF PLATES. 

Plate I. 

Gelatine needle cultures of the micrococci described. 

Fig. 1, Micrococcus a — Culture seven days old. 
"2 " b " " " 

"3, " c " six weeks old. 



i i 


4, 


i i 


D 


a ft 


k( 


5, 


a 


E 


Plate II. 



Drawings made from cover-glass preparations of urine cultures, 
•one day old, of the micrococci described. The preparations were dried, 
passed three times through a flame, and stained for a short time with 
alkaline-methylene-blue. The position of the germs in the field was 
determined by means of Abbe's camera lucida, Zeiss 2 mm. apochro- 
jnatic lens, No. 4 eye-piece. 

Fig. 1, Micrococcus A. 



( . 


2, 


a 


B. 


.( t 


3, 


a 


C. 


. i 


4, 


. < 


D. 


ii 


5, 


i t 


E. 



THE BACTERIOLOGICAL WORLD. 



359 



Plate II. 




<Hc*UlA l dLL^ 



Fig. 4 Fig. 5 

Proceedings of the American Society of Microscopists, 



360 THE BACTERIOLOGICAL WORLD. 

THE ETIOLOGY OF TETANUS. 

In the last number of the Annates de V Institut Pasteur, 
there appears ( from the Bacteriological Laboratory of Val-de- 
G-race ) a most interesting paper on tetanus, by Drs. Vaillard 
and Vincent, which appears to throw very considerable light 
on the subject of tetanus, and to clear up a number of points 
and observations that have hitherto been enshrouded in 
obscurity. After describing the organism, and identifying it 
with that already made familiar through the papers of recent 
writers, the authors give it as their firm opinion that, in cases 
of artificial inoculation of pure culture, it is always the 
poison introduced along with the bacillus, and not the organism 
itself, that acts upon the animal. This, indeed, seems to be 
probable, as they are able to prove that almost inconceivably 
minute doses of this poison, which they compare with snake 
poison, are quite sufficient to produce all the symptoms of 
most acute tetanus ; in fact, it was almost impossible, from 
some of the cultures that they obtained, to administer a dose 
that was not lethal. 

An exceedingly interesting feature brought out in the 
course of their work is that in no case was the poison devel- 
oped as soon as the organism began to grow ; in fact, gelatine 
cultures of the tetanus bacillus were never capable of pro- 
ducing toxic symptoms, until liquefaction of the gelatine had 
commenced, when spores were demonstrated to have been 
formed, and when the peculiarly disagreeable odor, so charac- 
teristic of tetanus cultures, had become perceptible. They 
associate both the odor and the peptonizing power with the 
formation of the poison in the cultures. That it was due 
merely to the presence of the spores that the material was 
poisonous, they demonstrated by heating their cultures to a 
temperature of 143.6° F. for a short time (a temperature 
which is quite incapable of interfering with the vitality of the 
spores), when it was found that cultures so heated and intro- 
duced by inoculation into a rabbit, or a guinea-pig, failed to 
produce any tetanus ; thus proving that, although the spores 
are not killed, the poison has been destroyed by the heat. The 
spores were proved to be living, by making fresh cultures from 
them in artificial media ; after a time they grew luxuriantly, 



THE BACTERIOLOGICAL WORLD. 361 

and, if left to grow eight or ten days, produced another crop 
of the poison. By simply washing away the poison from the 
spores with distilled water, they also obtained similar results, 
for, although the spores could still develop and form the specific 
poison in artificial media, they were, when inoculated, 
incapable of giving rise to any symptoms of tetanus. From 
the reaction to heat of a substance they were able to separate, 
and from its resemblance to the diastases in other respects, they 
conclude that they have obtained from tetanus cultures the 
true tetanus poison — a poison, however, that cannot be formed 
by the tetanus bacillus in healthy tissues. The micro- 
organisms are here so rapidly attacked by the leucocytes that 
they are rendered Jwrs cle combat before they have time to form 
their poison. 

It has long been well known that the tetanus bacillus could 
not develop in the tissues except, apparently, in the presence of 
other organisms, and the suggestion is offered, that these 
other organisms act in one of two ways : they either paralyze 
the activity of the leucocytes, or they draw off, as it were, their 
attention and activity from the tetanus bacillus, thus allowing 
it sufficient time to develop its characteristic products. It is 
interesting to note that Drs. Vaillard and Vincent consider 
that, in many respects, the tetanus bacillus is extremely like 
the diphtheria bacillus — the method of action on and in the 
organism being essentially the same in the two cases, the above 
factors, in all probability, playing a part in diphtheria, much 
as in the case of tetanus ; and it is evident that, in studying 
the one poison, much light may be thrown on the other. 
Behring and Kitasato appreciated this fact, and combined 
their forces to work out the question of immunity in these two 
diseases. It is obvious, however, from a consideration of some 
of the points that are indicated in this paper, that there are 
many sources of fallacy that will have to be eliminated before 
the ultimate explanation of the condition of immunity in 
protected animals can be given. The facts that this poison is 
active in such extraordinary minute quantities, and that the 
micro-organisms are able to grow with such difficulty in the 
human tissues, allow us to hope that extremely minute changes 
in the blood may be quite sufficient to secure the alteration 



362 THE BACTERIOLOGICAL WORLD. 

or breaking down of the virulent poison, even when it has 
become diffused throughout the system. So long as the 
organism is localized to the wound, there is, of course, more 
chance of coping successfully with the disease, although here, 
as in other diseases, there always appears to be a possibility of 
the poison exerting such a paralyzing influence on the cells that 
usually take up foreign substances, that secondary septic 
conditions may be liable to occur, even when the action of 
the tetanic poison can be antagonized so far as its primary 
effects on the cells are concerned. One question appears to be 
set at rest, and that is, as regards tetanus and diphtheria, the 
ptomaines have had their day, whatever may become of the 
products of other organisms. It may be accepted that here, 
at any rate, we have some subtle poison, which, although it 
has not yet been actually separated, has become so far isolated 
that it may be taken as proved, that it is not an alkaloid or 
basic poison. A most remarkable feature is, that, in pepton- 
izing gelatine with the filtrate from a meat-broth culture of 
the tetanus bacillus, the poisonous properties are lost, to a cer- 
tain degree, in direct proportion to the amount of gelatine that 
is peptonized ; this, taken in conjunction with the fact that 
the properties are not developed until the gelatine begins to 
liquefy, has led Drs. Vaillard and Vincent to suppose that the 
same agent that peptonizes the gelatine, is the active agent in 
bringing about the development of the toxic symptoms of 
tetanus. — The Lancet. 



THE REPUTED CAUSE OF INFLUENZA. 

Dr. Tezzier, of Lyons, France, claims that influenza is 
produced by a microbe, which he styles the strepto bacillus, 
whose habitat is putrid mud. That Russia is its home, is, in 
his opinion, due to the fact that bad drainage, filthy streets, 
and neglected barnyards are the rule — a condition particularly 
aggravated by swollen rivers and generally wide plains. — Med- 
ical and Surgical Reporter. 



THE BACTERIOLOGICAL WORLD. 363 

LUMINOUS MICROBES. 
Revue Scientifique, Paris, April 18. 

Mr. W. Beyerinck, whose interesting researches have already 
largely contributed to an acquaintance with the biology of 
luminous microbes, has continued his studies of them. In an 
important memoir, just published in the Archives Neerlan- 
daises des Sciences Exactes et Naturalles, he undertakes to 
show what are the elementary substances necessary for the 
activity of the light-giving function in the known luminous 
species. 

There are five species of light-giving microbes. First, there 
are the common luminous bacteria of the phosphorescent fish. 
Of these there are two species. Of the two the most lumin- 
ous is the photooacterium pfluegeri, which is also the most 
luminous microbe known ; the other is the photobacterium plios- 
phorescens, of which the light is not so strong. From a mor- 
phological point of view ph. phosphor escens has a form more or 
less spheroidal or oblong. The pfluegeri is longer. The lumi- 
nous bacteria of the Baltic are found in two varieties of one 
species ; the photobacterium fischeri which liquefies gelatine, 
and tYiQ ph. fischeri F. baltica, which does not liquefy gelatine. 
This is really the sole difference between the two varieties. 
Two allied species — completing the number of five — are the 
ph. indicum of the West Indian seas, and the ph. luminosum of 
the North Sea. These two species quickly liquefy gelatine. 

The luminous power of ph. indicum is great, and in that 
respect it ranks next to ph. phosphor escens. Like all the lumi- 
nous bacteria, ph. luminosum and ph. indicum are extremely 
sensitive to the presence of small quantities of sugar in their ali- 
ment. One per cent, of glucose or even less will completely 
extinguish the light-giving power of the luminosum, which 
with from three to five per cent, of glucose becomes incapable 
of liquefying gelatine, while larger doses kill it. The ph. indi- 
cum is a little less sensitive to glucose. It still continues to 
give light after the addition of four per cent, of glucose ; but 
under these conditions it takes on involved forms. 

M. Beyerinck gives the name auxanographie to the method 
which he employs to prove the influence of this or that ali- 
mentary- substance upon the different functions of microbes. 



364 THE BACTERIOLOGICAL WORLD. 

According to him the light-giving function, in the luminous 
bacteria, as in all the luminous creatures of the organic world, 
exists in living matter only. He has never succeeded in isolat- 
ing any luminous element or in making any light-giving matter 
become luminous, except in living cells. Mr. Eadzizewsky, 
however, is of a different opinion and likens organic phosphor- 
escence to chemical phosphorescence, produced by the combina- 
tion of certain chemical substances. 

A question put by Mr. Beyerinck is whether, in the strug- 
gle for existence, microbes derive any advantage from their 
light-giving function. He answers this question in the nega- 
tive. Observation so far does not warrant us in believing that 
the light emitted by microbes is in any way useful to those 
organisms. The light seems to be the accidental consequence 
of chemical transformations ; and this conclusion is corrobor- 
ated by the fact that ph. luminosum, for example, is much 
easier to obtain and preserve in a non-luminous than in a phos- 
phorescent condition. This microbe becomes luminous only 
at a low temperature towards 15° 6 miu. On the contrary, 
it is killed by insufficient nourishment and a temperature too 
low ; and it is in this condition that it is commonly found on 
the seashore. Other bacteria, ph. pfluegeri and phosphor escens 
are, it is true, less delicate. Nevertheless, when their carbon- 
ized aliment is very insufficient, their luminous power com- 
pletely disappears, so that in realty, these microbes pass the 
greater part of the year, in the sea and on the shore, without 
emitting any light. 

With the researches of Mr. Beyerinck on the light-produc- 
ing alimentation of luminous microbes, should be noticed the 
recent labors of Mr. 0, Gessard as to the influence of the nature 
and composition of nutritive matter upon the production of the 
pigments of the bacillus pyocyanique, the microbe which secretes 
a blue and green coloring matter. The production of the pig. 
ments is similar, in many respects, to the production of light. 
Both are produced under the condition of good alimentation 
and good health in the microbes, and testify to the appearance , 
under proper conditions, of a function, which of late years has 
been happily styled the function of luxury — since, in fact, this 
function, distinct from nutrition and multiplication, is indispen- 



THE BACTERIOLOGICAL WORLD. , 365 

sable neither to the life nor to the reproduction of micro-organ- 
isms. 

Mr. Gessard has discovered that the bacillus pyocyanic, in 
order to manufacture the two pigments which characterize it 
when in good health, has need of a special medium, the ordi- 
nary bouillon of cultures in the laboratory. Upon the albumen 
of an egg, the green pigment alone appears. This last does not 
appear in mediums formed of peptin and gelatine. Also it has 
been discovered that the addition to the nourishment of glucose, 
which kills the luminous microbes, stops the production of. the 
blue pigment characteristic of the pyocyanic bacillus. 

Moreover, as yet we know only vaguely what it is that 
makes microbes dangerous to human life ; but the most recent 
researches seem to agree in attributing this terrifying quality of 
microbes simply to an aptitude to multiply and reproduce them- 
selves. This dangerous quality of microbes appears to depend, 
however, on the quantity and quality of poisonous substances 
which it secretes, and this poison-producing function is also a 
very delicate function of luxury and dependent solely on favor- 
able conditions, which it is sometimes very difficult to unite. 
There . are, then, in microbes, three functions, very nearly 
related to each other ; these are the light-producing, the color- 
producing, and the poison-producing. Light, color and poison 
are three functions superadded to the indispensable functions of 
life, and which appear to be dependent on conditions which, if 
not identical are at least nearly related to each other from" a 
general point of view. The study of one of these functions may 
help acquaintance with the others ; and if we can manage to 
understand thoroughly the conditions under which poison-pro- 
ducing microbes exercise their peculiar function, we shall soon 
be able to stop the exercise of that function ; that is to say, we 
shall soon be able to cure the infectious maladies. —Literary 
Digest. 



CHEMICAL REACTIONS OF KOCH'S TUBERCULIN. 

In the Farmatzevtitchesfcy Juernal, No. 9, 1891, p. 136, 
Mr. I, Gertel, a chemist^ publishes the result of his examina- 
tion of a specimen of Koch's "tuberculin." The "lymph" 



366 THE BACTERIOLOGICAL WORLD. 

forms a cinnamon-brown, odorless, viscid fluid, sp. gravity of 
1.17 and of a neutral reaction. Concentrated sulphuric acid 
dissolves it easily and completely, the solution being at first 
colorless, but after a while assuming a cinnamon-brown color. 
Concentrated hydrochloric acid does not manifest any action on 
the lymph, while concentrated nitric acid produces a slight 
opacity, appearing at the point of contact between the two 
fluids. When immersed in a sulphuric acid solution of tuber- 
culin, crystals of bichromate of potassium soon become covered 
with bubbles ( which are due to glycerine being present in the 
preparation). The following reagents give considerable pre- 
cipitates on being added to a 10 per cent, aqueous solution of 
tuberculin : Tannic acid, picric acid, chloride of gold, nitrate 
of silver, sulphate of copper, nitrate of mercuric suboxide ; but 
chloride of platinum, neutral acetate of lead, and chloride of 
tin, produce a comparatively slight precipitation ; iodide of 
potassium and cadmium ( double salt) give a distinct turbidity, 
but no actual deposit. The following substances behave entirely 
negatively toward the solution : Iodide of potassium and mer- 
cury (double salt), bi-iodide of potassium, chloride o£ iron, 
ferro-cyanide of potassium, chloride of mercury, molibdenate 
of ammonia, rhodanide of potassium, chromate of potassium, 
ammonia, and sulphate of iron. When heated with acids, the 
aqueous solution assumes an intense yellow color. Strongly 
diluted aqueous solutions give voluminous precipitates with 
alcohol, the deposits being redissolved on a free addition of 
water. A 10 per cent, aqueous solution, as well as more diluted 
ones, when kept at the ordinary room temperature, becomes 
turbid even on the next day. As to the lymph itself ( that is, 
an undiluted preparation ), it has not undergone any changes 
even after four months' keeping. — Medical and Surgical 
Reporter. 



SUMMARY OF THE RIPENING OF CREAM. 

BY H. W. CONN, PH. D. 

The most important points brought out in the present 
article are the following : 

The ripening of cream is a process produced by bacteria 
growth. This is shown by the immense number of bacteria in 



THE BACTERIOLOGICAL WORLD. 367 

the cream, and their almost incredible multiplication daring 
the ripening process, and by the fact that all conditions which 
hasten bacteria growth also hasten the ripening process. 

The variety of bacteria which are found growing rapidly in 
ripening cream is very great, and there is no uniformity in 
regard to species. Hardly two specimens of cream in the same 
creamery are ripened by the same variety of bacteria. The 
conditions affecting the growth of different species of bacteria 
are entirely beyond the control of the butter maker, and it is 
impossible to obtain uniformity of results by the ordinary 
methods of ripening. This fact is doubtless the cause of the 
occurrence of butter of inferior quality at certain seasons, and 
is one of the causes of difference between the butter of different 
creameries and dairies. 

The bacteria which ordinarily ripen cream produce three 
classes of effects upon it during their growth. Some of them 
produce lactic acid, or small quantities of other acids. Some 
of them produce a rennet-like ferment which tends to pre- 
cipitate the casein, and also a trypsin-like ferment which digests 
or peptonizes the proteid matters present. Some of them pro- 
duce no effect beyond a slight decomposition and the possible 
evolution of volatile products. All of them assist in giving the 
peculiar flavor to the cream, and therefore probably the aroma 
to the butter. Perhaps a satisfactory aroma can be obtained by 
the use of single species if carefully selected. 

No especially characteristic species have been found con- 
nected with the ripening process, The acid-forming species 
are, however, the most uniformly present in great numbers, 
and these are probaby the most intimately associated with the 
ripening. — Storr's School Experiment Station Report, 1890 -1891. 



THE MICROCOCCUS OF BITTER MILK. 

BY H. W. CONN, PH. D. 

* 4J % i£ 

The organism which I have found in the specimen of bitter 
cream is undoubtedly a different one from that found by Weig- 
mann. The following is a description of its characters : 



368 THE BACTERIOLOGICAL WORLD. 

Morphology. — It is a good-sized micrococcus which has no ten- 
dency to form chains. Occasionally two may cling together, produc- 
ing diplococci. This is especially the case in gelatine and milk cultures. 
In agar it becomes longer, and rods are found several times as long as 
broad, which I have not been able to break up into cocci . 

Mobility. — Not mobile. 

Relation to the Air — Aerobic. — Under a mica plate it grows very 
slowly, giving rise to bubbles of gas, and the liquefaction of the gela- 
tine takes place slowly. No such bubbles are produced where the 
plate is not covered with mica, and liquefaction is rapid. 

Color.— White. 

Colony on Gelatine. — Under the surface the colony is smooth and 
round. As it approaches the surface it becomes almost immediately 
diffused over the gelatine into a fine, thin granular mass, which 
spreads rapidly. There is no definite shape to the colonies after this. 

Gelatine Stab Culture. — At first a very shallow pit is formed, but 
this spreads into a uniform layer of liquefying gelatine, which slowly 
deepens until the whole is liquefied. The resulting liquid is very 
slimy. 

Agar- Agar. — A white glistening uniform growth. 

Potato. — A white mass of more or less discrete bunches, glistening 
and moist ; color of the potato not changed. 

Milk. — Curdled in one day in a warm oven. Becomes very bitter. 
The litmus solution which is added to the milk turns slightly red . 
Later the color of the litmus is bleached. The curd which is formed 
soon begins to dissolve, though the solution is never quite complete. 
The liquid thus formed is slimy. 

Bouillon. — Growth abundant. A thin membrane forms on the 
surface. The bouillon becomes remarkably slimy, and can be drawn 
out into threads ten feet long. Odor and taste like that of bitter 
milk. t 

The most remarkable feature of this organism is the power 
of rendering bouillon and gelatine slimy. The tenacity of the 
slime is hardly credible. The mass can be sometimes drawn 
out into threads ten feet long and no larger than a thread of 
silk, so as to be hardly visible. Curiously enough, though both 
bouillon and gelatine are thus rendered slimy, the sliminess 
does not at first appear in milk. After the digestion of the 
curd has taken place the liquid becomes slimy, but no slimy 
milk proper is formed ; so this organism is, therefore, not one of 
those which produce the well-known slimy milk. 

The taste produced in the milk is extremely bitter. It is 
not exactly like that of the original bitter cream from which 



THE BACTERIOLOGICAL WORLD. 369 

the organism was obtained. This is due, however, to the fact 
that other organisms were present in the cream. I found that 
if a lot of sterilized milk was inoculated with the bitter organism 
and at the same time with one of the lactic acid-forming species, 
the taste is almost exactly that of the original cream. 

This organism undoubtedly produced a chemical ferment 
or enzyme, with action similar to that of rennet. 



A PIGMENT-PRODUCING BACTERIUM. 

At a recent meeting of the Amsterdam Royal Academy of 
Sciences Beyerinck -read a paper on the life history of a pig- 
ment bacterium. This organism, bacillus cyaneo-fuscus, is the 
cause of a much-feared local coloring process in Dutch cheese ; 
called "blue illness"; and of "black glue," a calamity 
observed in a gelatin factory. The natural habitat of the organ- 
ism is ditch-water and ground- water. It is a peptone bacterium, 
i. e., it can be fed with albuminous matter alone. Hence a 
one-half per cent, solution of peptone in water is sufficient 
nutriment ; gelatin, egg albumen, fibrin and casein are also, 
each alone, sufficient, but they are peptonized by a secretion of 
the bacterium, before absorption. The bacterium produces 
two pigments — one, deep blue spherites ; the second, a dark 
brown diffusible coloring-matter. On this account a peptone 
solution acted upon by the organism becomes quite black, — 
Exchange. 



AN ALKALOID FROM TUBERCLE BACILLI. 

Zuelzer, the discoverer of sepsin, announces ( Berl. Klin, 
Woch.) that, by a very simple method, he has succeeded in 
separating a ptomaine from the metabolic products of tuber- 
cle bacilli. For this purpose the entire contents of culture- 
tubes were repeatedily extracted by means of hot water acidified 
with hydrochloric acid, filtered, evaporated, decomposed sev- 
eral times with sulphuretted hydrogen, again filtered, and 
finally dried. In this way was obtained an almost white crys- 
tallized salt, easily soluble in hot water ( 104° F.), but not in 



370 THE BACTERIOLOGICAL WORLD. 

cold. The solution had a slight yellow tinge. The dried salt, 
when kept for some time, turned to a clear brown. Its toxic 
effects upon rabbits and guinea-pigs were strongly marked and 
very characteristic. — Druggists' Circular. 



VACCINATION AS A PREVENTIVE OF INFLUENZA. 

Dr. G-oldschmidt of Madeira hopes to conquer influenza by 
means of animal lymph, as re-vaccination against small-pox 
seems to confer almost perfect immunity from influenza. In 
consequence of the neglect of vaccination small -pox prevails 
frequently in Madeira, and not long ago it carried off more than 
a thousand victims. The doctor went in for re vaccination, 
and now finds that not one patient out of one hundred and 
twelve successful cases suffered from influenza, and out of 
ninety-eight successfully vaccinated, only fifteen persons were 
seized by influenza, and these in each case but very slightly. 
Kecently vaccinated infants did not catch the influenza. It 
therefore appears to follow that re.vaccination gives effective 
protection, not only against small-pox, but also against influenza. 

EXAMINATION OF INFUSORIA. 

Every disciple of the microscope knows the difficulty of 
observing accurately these lively denizens of the water. If a 
drop of thick water solution of cherry gum be put into the live- 
cage, it has the effect of retarding the movements of the creat- 
ure without materially affecting the ciliary motion. 

EXAMINATION OF DRINKING WATER. 

The water should be filtered through a short column of 
fine sand in the stem of a funnel, the sand being supported on 
a plug of wire-cloth placed beneath it. The sand retains the 
whole of the organisms contained in the water. The sand is 
then washed with distilled water, both being allowed to run 
into a test-tube. The mixed sand and water, after being 
shaken, must be left for the sand to sink, leaving the organisms 
suspended in the water. A definite quantity of this can be 
taken out by a pipette and the organisms enumerated. — Inter- 
national Journal of Microscopy & Natural Sciences. 



THE BACTERIOLOGICAL WORLD. 371 

MISCELL A.NJEO US. 



BACTERIOLOGICAL INVESTIGATIONS CONCERNING THE 
ETIOLOGY OF PNEUMONIA IN CHILDREN. 



Neumann {Rev. Mens, cles Mai. del' Enf., July, ]890) 
says : 

"Bacterial investigations in a number of cases of pneumo- 
nia in children led the author to the conclusion that the pneu- 
mococcus of Fraenkel is the unvarying cause of fibrinous 
pneumonia in children, the same as it is in adults. The same 
organism is also to be found in most of the cases of broncho- 
pneumonia in children. In sixteen cases of broncho-pneumonia, 
nine of which were consecutive to measles, the pneumococcus 
was found ten times. It is often very difficult to find the 
pneumococcus in broncho-pneumonia on account of its frequent 
association with other micro-organisms, including the strepto- 
coccus aureus et albus, streptococcus pyogenes, proteus vulgaris, 
and bacillus pyocyaneus. If the pneumococcus is not found, 
one must remember that it was present at the beginning of the 
disease, but has become attenuated, and has disappeared as the 
disease has progressed. The presence of an organism is an 
evidence of the existence of pneumonia, but the absence of the 
organism does not necessarily indicate that pneumonia is not 
present. Prophylaxis against pneumonic infection should be 
carried out by means of thorough ventilation and cleanliness of 
the apartments occupied by children with pneumonia, also by 
disinfection of the buccal cavity, and calling attention to the 
danger residing in the sputa of patients with pneumonia." — 
Arch. Pediatrics. — Gin. Lancet Clinic. 



372 THE BACTERIOLOGICAL WORLD. 

TECHNIQ UE. 



New and rapid method of staining the capsule of 
bacillus pneumoniae. — Dr. U, Gabbi employs the following 
method for staining the capsule of bacillus pneumonice Fraenkel. 
The sputum to be examined is spread upon a cover-glass and 
quickly dried in a spirit lamp flame. Two or three drops of a 
solution of 2.5 grams carbolic acid, 1 gram fuchsin, and 15 
grams alcohol, in 100 grams distilled water, are then dropped 
on the preparation, where they are allowed to remain for one 
minute. It is then quickly washed in water, and in prepara- 
tions thus treated the bacillus is stained dark red and its 
capsule bright red. The staining of the capsule vanishes if 
left too long in the water. — Journal of the Royal Microscopical 
Society, — Microscopic Bulletin. 

"Air-gas'' for bacteriological work. — Dr. 0. Katz, 
who had to work on Rodd Island, N. S. Wales, where the ordi- 
nary appliances of civilization were not available, made use of 
the "Alpha patent gas-making machine." This apparatus 
produces gas in the shape of a mixture of atmospheric air and 
the vapor of petroleum spirit (gasoline), the mixture being 
called air-gas. By means of weights atmospheric air is pumped 
through a drum into a chamber, where it is impregnated with 
the vapor of the volatile fluid. The mixture then passes into a 
gasometer, from which the burners are supplied automatically. 
The author used a 40-light machine, capable of yielding 200 
feet of gas an hour. 

The author considers that air-gas makes an efficient substi- 
tute for coal-gas for ordinary lighting purposes, and he also 
used it with success for heating thermostats, and also for 
other bacteriological purposes. — Journal Royal Microscopical 
Society. 

Negative indol-reaction as a test for the typhoid 
bacillus. — The resemblance, from their morphological and 
cultivation aspects, of numerous bacilli to the bacillus of 
typhoid, suggests the importance of having some specific test 
which may be easily applied in making a differential diagnosis 
between the bacillus of typhoid and other bacteria. 



THE BACTERIOLOGICAL WORLD. 373 

Recent. observations have shown that potato cultivations do 
not afford a certain criterion ; but after experimenting with six- 
teen kinds of bacteria, Dr. S. Kitasato found that these, when 
cultivated in bouillon, produced indol, while the typhoid 
bacillus did not. 

The indol was tested for by Salkowski's method, that is, by 
treating the cultivations with a solution of nitrate of potash, 
and then adding a few drops of strong sulphuric acid. With 
this test the typhoid bacillus remained colorless, while the 
other sixteen bacilli assumed the characteristic red hue. 

Careful chemical analysis also constantly showed the absence 
of indol from the typhoid cultivations. 

As all these pseudo-typhoid bacilli developed on potato 
quite differently from the real typhoid bacillus, the author con- 
cludes that the negative indol reaction is in itself no better test 
than the growth on potato is. — Journal of the Royal Microscop- 
ical Society. 

On bacteriological examination of the sputa. — As 
is well known, all methods of a bacteriological examination of 
the sputa, as yet proposed, are applicable only to a newly 
expectorated discharge, and not to one of old standing. The 
inconvenience arising from the circumstance, for a busy practi- 
tioner (especially for a country one), being very considerable, 
Dr. Wikhail Y. Saveliefd, of Zemlansk, draws attention (Medit- 
einshoie Obozrenie, No. 21, 1890, p. 835), to a simple method 
for preserving the sputa, and examining it bacteriologically, at 
a more or less remote date after its expectoration. The author 
makes the patient expectorate directly into a vessel containing 
95 per cent, alcohol, in which medium the discharge remains 
unchanged for five to six months, and even longer. Since the 
vehicle causes the coagulation of the sputum, the clots should 
be liquefied by treating them with a two per cent, aqueous 
solution of caustic potassa, before the microscopical examina- 
tion. The following technical details are recommended by the 
writer : A small lump is taken out ( by means of two steril- 
ized needles, and with the usual bacteriological precautions) 
from the middle of a clot and placed on a cover-glass, after 
which two or three drops of the potassa solution are poured 
over the bit (from a pipette). After two or three seconds, the 



374 THE BACTERIOLOGICAL WORLD. 

specimen is covered with another glass slip, the whole com- 
pressed with the fingers between the bibulous paper, and the 
sputum, stained after this or that method in vogue. — Valerius 
Idelson, in St. Louis Medical and Surgical Journal. 

A NEW TEST FOR ALBUMEN AND OTHER PROTEIDS.* — Dr. 

John A. Mc William gives, in The British Medical Journal, a 
method of testing for proteids by salicyi-sulphonic acid. Take 
a small amount of urine (for example, 20 minims), preferably 
in a very small test-tube, and add a drop or two of a saturated 
watery solution of the reagent. If the urine is strongly alka- 
line, an extra drop or two of the acid should be added, and, if 
no opalescence or precipitate occurs, it is well to test the reac- 
tion with litmus paper, and make sure that the urine has been 
rendered strongly acid. On adding the reagent, shake the tube 
quickly, so as to mix its contents. Then examine at once. 
The occurrence of an opalescence, or cloudiness, immediately or 
within a very few seconds (for example, 2 to 3 seconds), is a 
test for proteids, intermediate in delicacy between the cold 
nitric acid test on the one hand, and the acetic acid and heat test 
(in favorable circumstances) on the other. The development 
of an opalescence some time after ( for example, -J- to 2 
minutes), is a more delicate test than even acetic acid and heat, 
and shows the presence of minute traces of proteid, which are 
probably insignificant from a clinical point of view, as a rule. 

Next, heat the tube to the boiling point. If the precipi- 
tate, or opalescence, is caused by the ordinary " albumen " 
(albumen and globulin), commonly present in albuminous 
urine, it does not disappear on heating, but, on the other hand, 
becomes markedly flocculent. But, if the precipitate, or opal- 
escence, is due to the presence of albumoses or peptones, it 
clears up on heating ( before the boiling point is reached), and 
reappears when the tube cools. 

The phenyl-hydrazik test for sugar. — Havilburg 
(Centralbl. fur klin, Medicin, January 31, 1891), of Rio de 
Janeiro, proposes a modification of the Fischer-Jaksch test for 
the presence of sugar in the urine, which he considers more 



* These matters, though not truly bacteriological, concern the 
mioroscopist and bacteriologist. 



THE BACTERIOLOGICAL WORLD. 375 

reliable. Two parts of phenyl-hydrazin hydrochlorate and 
three of sodium acetate are added to half a test-tube full of 
water, to which a like quantity of urine to be tested is added. 
Instead of boiling over a water-bath, the mixture is shaken 
with chloroform. In a little time the fluids separate. If sugar 
be present, canary-yellow crystals appear in the upper layer. 
The reaction is more striking if the fluid is boiled and per- 
mitted to cool before the addition of chloroform. Havilburg 
suggests that the procedure might be adapted for use as 'a ready 
means of quantitative determination of the amount of sugar 
present. — American Journal Medical Science. 

KlJEHKE'S METHYLENES LUE METHOD OP STAINING: BAC- 
TERIA. — This method is especially recommended for staining 
bacteria in sections of animal tissues, although it is equally 
applicable to cover-glass preparations made from fresh tissues. 
The usual differences in the method of staining cover- glass 
preparations and sections are to be observed. 

The advantages to be derived from this method are found 
in its being applicable to all known forms of bacteria. It 
eliminates the use of special stains for certain micro-organisms 
where only their presence is to be demonstrated. It possesses 
superior powers of differentiation between bacteria and the 
tissue elements. The method as given by Dr. Kuehne is 
essentially as follows : 

The sections which have been cut by the ordinary method 
( although Dr. Kuehne recommends the freezing microtome for 
this purpose) are transferred directly from alcohol to a watch- 
glass containing carbol-methylene-blue. First. The sections 
should remain in this staining fluid for about half an hour ; 
some bacteria, such as the bacillus of leprosy, requiring a longer 
time, one to two hours. If the sections remain in the staining 
fluid for a much longer period, the differentiation between the 
germs and tissue elements becomes more difficult. 

After staining for the desired length of time, the exact 
period of which will have to be determined by test experiments 
for the different germs and tissues, the sections are rinsed in 
clear water and then placed in acidulated water, second, until 
they become a pale blue. They are then washed in a weak 
watery solution of carbonate of lithium, third, and again placed 



376 THE BACTERIOLOGICAL WORLD. 

in clear water. This part of the procedure is very important, 
and to insure good results should be performed with much care. 
The time that the sections should remain in the decolorizing 
agents varies with their thickness, histological structure, and 
the intensity of the stain, making it impossible to give any 
definite rule to be followed. The degree of decolorization can 
be very nearly determined at any moment by moving the sections 
about in the fluid by means of a glass rod. If the section is 
very thin, or if there are other reasons why it should take up 
very little of the stain, a momentary immersion in the acidulated 
water is sufficient. In all cases where the staining process is 
completed the sections should have a pale blue color, for if 
darker the over-stained corpuscles and cell-nuclei of the tissue 
would obscure the bacteria. In Cases where it is feared that too 
much color has been removed in the acid a drop of a saturated 
watery solution of methylene-blue should be added to the 
lithium-water. 

After the sections have remained in the water for some 
minutes they are dehydrated in absolute alcohol in which, in 
difficult cases, a little methylene-blue may be dissolved, and 
then transferred to a watch-glass containing methylene-blue 
anilin oil ; fourth, the sections can be dehydrated in the alcohol 
without injury to the stained bacteria. The sections are now 
transferred to pure anilin oil., in which they are rinsed and then 
placed in some essential oil, as turpentine, where they should 
remain for two minutes. In order that the sections should be 
perfectly cleared they are transferred from the turpentine to 
xylol, from which they are mounted in balsam. It is recom- 
mended that the sections should pass successively through two 
xylol baths in order to secure absolute elimination of the anilin 
oil. The xylol may be used for a considerable number of 
sections. 

Dr. Kuehne employs a glass rod for transferring the sections 
from one solution to another instead of the ordinary spatula or 
section-lifter. The end of a small glass rod is immersed in the 
fluid containing the section, which is allowed to fold itself over 
the rod, and in this position it is lifted from the fluid. The 
end of the rod is then gently immersed in the second liquid, 
where the section unfolds itself from the rod and floats upon 



THE BACTERIOLOGICAL WORLD. 377 

the surface. In this way the danger of tearing the section is 
diminished and the time required for their transfer from solu- 
tion to solution is much shortened. This is an important con- 
sideration where a large number of sections are to be stained. 

First. Carbol-meihylene-blue. — This is prepared by grind- 
ing in a mortar 1.5 grams of methylene-blue with 10 ccm. of 
absolute alcohol until dissolved ; 100 ccm. of 5 per cent, car- 
bolic acid are gradually added and thoroughly mixed with the 
alcoholic solution. The resulting liquid is preserved in a well- 
stoppered bottle, until used. When only a small quantity is to 
be employed it is better to prepare only a half, or a quarter 
even, of the above quantity, as its staining power is diminished 
by long standing. It should always be filtered before using. 

Second. Weak acidulated water. — To 500 ccm. of distilled 
water add 10 drops of nitric acid. 

Third. Lithium-water. — To 10 ccm. of distilled water 
add from 6 to 8 drops of a saturated watery solution of carbonate 
of lithium. The saturated solution may be used as a decoloriz- 
ing agent in sections with over-stained nuclei. 

Fourth. Methylene-blue anilin oil. — About one-half gram of 
methylene-blue is ground in a mortar with 10 ccm. of pure 
anilin oil. When the oil is saturated with the coloring matter 
the entire mass is poured unfiltered into a vial, where the 
undissolved coloring matter will settle, leaving the saturated 
supernatant oil clear. To a watch-glass of pure anilin oil add 
a few drops of the saturated methylene- blue- oil until the 
desired degree of colorization is obtained. 

Bacteriological examination of influenza. — M. 
Kirchner ( Zeitschrift f. Hygiene, Bd. 9, Heft, 3) has made an 
examination of the bacteria in the sputa and blood of patients 
suffering from influenza, and, from the thoroughness of the 
work and the uniformity of the results, his observations appear 
to be very trustworthy. The examinations were made upon 
thirty patients in the military garrison in Hanover, Germany. 
Cultures were made in all cases where it was possible, but cir- 
cumstances prevented the attempt to cultivate organisms in 
some cases. Still the microscopic examination in all and the 
cultures in the cases where these were made showed the con- 
stant presence of an organism with distinct characteristics. 



378 THE BACTERIOLOGICAL WORLD. 

This organism was the only one found in the blood and the 
only one which was found in every case in the sputa. It is a 
diplococcus much smaller than Fraenkel's diplococcus pneu- 
monia (Sternberg's micrococcus Pasteuri ), and the distance 
between the two cocci is greater in the former than in the latter. 
The latter has somewhat pointed ends, whereas Kirchner's 
diplococcus has perfectly rounded ends. Kirchner's organism 
grows much more vigorously on agar than Fraenkel's, though 
like the latter it only grows at a high temperature, but does 
not die out as rapidly. The experiments upon animals are not 
yet complete ; but where a culture of the organism was injected 
into the thoracic cavity of a guinea-pig, the animal died in 
twenty-four hours, and the organisms were found in the lungs. 
Inoculations under the skin had no effect. The diplococcus, 
judging from these experiments, seems not to have special 
pathogenic properties. 

[ Although Kirchner's observations make it probable that 
the diplococcus described above is the cause of epidemic influ- 
enza, one of Koch's postulates is unfulfilled, viz., the produc- 
tion of influenza or a similar disease by inoculating pure 
cultures in animals ; maybe this test is impossible, for, as in 
several other cases, it is possible that lower animals are not sus- 
ceptible to influenza. This makes the proof that the organism 
is the cause of the disease much more difficult, and necessarily 
leaves the matter in doubt.] 

Disinfection. — The question of disinfection would seem 
to be one which had long ago been exhausted, and yet Behring 
( Desinfection, Desinfectionsmittel und Desmfectionsmethoden, 
ibid.) has published the results of experiments carried on in the 
Hygienic Institute in Berlin, which show that in much of the 
work done under this head many important conditions were 
lost sight of. It will not be possible in this review to give 
even a meager outline of Behring's experiments, but the 
original article is worthy of careful study, as it is a most thorough 
treatment of the subject. 

Behring points out that in all experiments on disinfection 
the following points must be taken into consideration : 



THE BACTERIOLOGICAL WORLD. 379 

"lirst. The unquestionable proof [ in any given case ] 
that the disinfection has been accomplished, i. e., the real kill- 
ing of the organisms [not merely the prevention of growth ]. 

"Second. The chemical character of the material to be 
disinfected. 

"Third. The kind of bacteria. 

"Fourth. The length of time that the disinfectant works. 

"Fifth. The temperature at which the disinfectant is made 
to work. 

"Sixth. The number of bacteria." 

Behring shows that the above points have not all been con- 
sidered, as a rule, in previous experiments with disinfectants, 
and hence are faulty. — The Brooklyn Medical Journal. 

Diagnostic examination. — Dr. Laveran gives a very 
clear account of his methods of examination of the blood in 
cases of malaria. He points out that such examination is 
exceedingly necessary in hot countries, where typhoid fever or 
sunstroke may be mistaken for malaria, or vice versa. An 
examination of the blood always puts the matter beyond doubt. 
He recommends that the examination should be made just at 
the beginning of a febrile attack, and before quinine has been 
administered, as during the period of apyrexia the organisms 
are seldom found in the peripheral circulation, but appear to 
be collected in the internal organs, and especially in the spleen. 
For the examination of the fresh blood, the skin should be 
cleansed with soap and water, rinsed with alcohol and carefully 
dried, then, everything' being ready, the finger is pricked with 
a pin that has been heated to redness, and allowed to cool, the 
little round globule of blood that appears is touched with a 
clean slide ; a cover glass is lowered down onto the blood, which 
is pressed out until the film assumes a transparent yellow color ; 
the film is then not too thick, and should be examined at once. 
The clot that is formed at the margin prevents the drying of 
the film ; but, in order to keep the film thin, it is better to wipe 
away the blood that is pressed from under the cover glass, and 
then to surround with paraffine. Daylight and no sub-stage con- 
denser should be used for examinations, or the organisms are 
rendered too transparent. The movements of the flagelia and 



380 THE BACTERIOLOGICAL WORLD. 

the amoeboid movements can all be made out. If the organisms 
are pigmented they are readily enough seen, but a most careful 
search may have to be made for those non-pigmented organisms 
that sometimes adhere to the red blood corpuscles. If the speci- 
men is to be preserved for further examination the film should 
be prepared by compressing between two cover glasses, which 
are carefully separated, allowed to dry, and passed two or three 
times through a clear flame ; each film is mounted unstained 
and dry, with a parafrme rim to keep out the air, and to retain 
the cover glasses in position. When it is wished to stain the 
organisms in order to bring them into special prominence, the 
films, after being heated on the cover glass, are put into a 
mixture of acohol and ether; they are then allowed to dry, after 
which they are stained with a concentrated aqueous solution of 
methylene-blue for thirty seconds ; they are then rinsed in 
water and mounted dry, the cover glass being surrounded with 
paraffin. The leucocytes are colored deep blue, the free 
spherical organisms and those adhering to the red blood cor- 
puscles pale blue, whilst other forms are scarcely tinged. 
A contrast stain may be obtained by using eosin. With these 
stained preparations artificial light may of course be used. In 
all cases where possible both methods of preparation should be 
resorted to, as each has its advantages. — British Medical Jour- 
nal. 

Bacilli in syrhilis. — Dr. von Marschalko, who has been 
working in the pathological laboratory of the Buda-Pesth Univer- 
sity, at the subject of the bacteriology 61 syphilis, has devised a 
method of staining, by which bacilli may be found in syphilitic 
growths of various kinds — such as papules on the skin, the indura- 
ted base of a chancre, and condylomata, as well as in the secretion 
of primary sores, while he has not been able to find them in 
non-syphilitic growths, or in the secretion of soft chancres. 
Whether they are really inseparably connected with syphilis, 
can, of course, only be decided by a prolonged series of observa- 
tions, but there would at least appear to have been a prima- 
facie case made out for further investigation. The method 
employed is by double staining, as follows : The object, after 
having been hardened in alcohol, is immersed for thirty-six 



THE BACTERIOLOGICAL WOULD. 381 

hours in an alkaline solution of methyl blue ; it is then rinsed 
well with water, and restained by a concentrated solution of 
vesuvin in water. The bacilli are stained blue, and can be 
easily distinguished from the tissues, which are brown. — 
Medical and Surgical Reporter. 



REMEDIES AND CLINICAL NOTES. 



HERPES GENITALIS. 

I£ Acid. Carb ni. xxx. 

Tarro-Petrolene 5 ij. 

M. ft. ungt. Sig. : Apply two or three times, daily. 

Note : Pyoktanine may be added in the quantity of, say, gr. v. or 
substituted for carbolic acid in the dose of ten grains. 

SUMMER DISTURBANCES OF CHILDREN. 

In fermentative disorders of the alimentary canal in the young, 
middle-aged or old, Listerine has given most satisfactory results. In 
the summer diarrhoea of children, Dr. I. N. Love, of St. Louis, speaks 
very highly of it, given in combination with glycerine and simple 
syrup. A formula that I have time and again used — in fact, it has 
almost become routine with me of late years — is as follows : 

]$. Bismuth Sub. Nit ' half a drachm. 

Tr. Opii twenty drops. 

Syr. Ipecac 

Syr. Rhei Arom aa two drachms. 

Listerine half an ounce. 

Mist. Creta one ounce. 

M. Sig. — Teaspoonful as often as necessary, but not more fre- 
quently than every three or four hours. This for children about ten 
or twelve months old. 

D. J. Roberts, M. D., in Southern Practitioner. 



382 THE BACTERIOLOGICAL WORLD. 

QUERIES AND ANSWERS. 



Mr. Editor. — Which speedy permanent treatment for rebellious, 
syphilitic ulceration on nose and upper lip do you advise? 

A Subscriber. 

A . This is not a question properly belonging to bacteriology and. 
the editor is not a specialist in skin diseases. However, a competent 
physician to whom this question was submitted says : Use Iodide of 
Potassium internally at the highest possible dose and apply the follow- 
ing locally and constantly : 

1$. Hydrargyri Chlorid. Corrosiv gr. ij. 

Aristol 3 j- 

Tarro-Petrolene 5 1 J- 

M. ft. oint. Sig. : Apply two or three times daily after one good 
scraping of surface of ulcers and drying. 

J. B. W., M. D. 



LABORATORY NOTES. 



SECOND REPORT. 

Dear Sir : — Since the opening of the Chicago Pasteur Institute 
(July 2, 1890) 55 persons have received treatment, 

Thirty-one came from Illinois — 10 from Iowa — 4 from Indiana — 2 
from Ohio — 2 from Minnesota — 1 from Missouri — 1 from Tennessee — 2 
from Kentucky — 1 from South Dakota — 1 from Arizona. 

Thirty-six were male and nineteen female. 

The youngest patient was 2£ years old and the oldest 56 . Fifty- 
one were bitten by dogs, three by cats and one by a skunk. 

Thirty-one persons were bitten in the hands and arms, three in the 
face, and twenty in the body and legs. Twenty-four had their 
wounds cauterized with chemical agents, and thirty-one had not. 

Thirty-three persons were bitten by animals recognized and 
ascertained of being rabid, by experiments made upon rabbits, by the 
death of persons and other animals bitten by the same, or by symptoms 
shown during life ; and twenty-two persons were bitten by animals 
strongly suspected of being rabid . 

All the persons treated are now enjoying good health. 

Adhering to the principles of strict and careful sterilization of all 
instruments and fluids used, no abscesses nor inflammations nor even 



THE BACTERIOLOGICAL WORLD. 383 

irritation of any kind occurred at the seat of inoculations, thus testing 
the purity of the material used. 

It has been made a practice at this institute never to subniit per- 
sons to treatment, unless in need of it ; consequently ninety-six persons 
were refused treatment as not reasonable evidence was produced to 
demonstrate the madness of the animals ; all these persons are also in 
good health confirming the correctness of our judgment. 

A. Lagorio, M. D. 

Chicago, May 1, 1891. Director. 



READING NOTICE. 

The Bacteriological World takes pleasure in drawing attention 
to the following very important announcement : 

We are gratified to inform our readers that arrangements have 
just been completed, and contracts signed, by which we have been 
appointed sole agents for the United States and Canada for the sale of 
the famous microscopes and objectives of Charles Reichert, of Vienna. 
His objectives stand in the very front rank. His microscopes are 
eminently esteemed in the laboratories of Europe. Further announce- 
ments in regard to catalogues, etc., will be made in due course. 

Respectfully, 

James W. Queen & Co. 



LITERATURE. 

THE ARENA. 

CONTENTS FOR JUNE NUMBER. 

The Unknown .Camille Flammarion. 

The Logic of Port Royal. ... Prof. Brentano. 

A Radical Remedy Prof. Joseph R. Buchanan. 

Through the Slums of Boston By the Editor. 

The Governmental Control of Railways. C. Wood Davis. 

What is Buddhism? Charles Schroder. 

The Irrigation Problem in the North- 
west James Realf , Jr . 

Evolution and Christianity Prof. Jas. T. Bixby, Ph. D. 

The Realistic Trend of Modern German 

Literature Emil Blum, Ph. D. 

At Home Amelia B. Edwards. 

Faith in God as a Personal Equation. . .Rev. C. A. Bartol. 

The Immigration Problem Rabbi Schindler and others 

The Malungeons Will Allen Dromgoole. 



384 THE BACTERIOLOGICAL WORLD. 

The Dangers of an Irresponsible Educa- 
ted Class in a Republic Helen M. Gardener. 

The Chivalry of the Press Julius Chambers. 

Working-Girls' Clubs Helen Campbell. 

The New Old Testament ..Rev. John W, Chadvvick. 

LIPPINCOTT"S MONTHLY MAGAZINE. 

CONTENTS OF MAY NUMBER. 

Vampires Julien Gordon. 

The Experiences of a Photographer. . . . A. Bogardns. 

Lost Treasures of Literature William Shepard. 

Poems Charles Henry Luders. 

That Hound o' Joel Trout's M. G. McClelland. 

Absence Owen Wister. 

Some Familiar Letters by Horace Gree- 
ley — III Edited by Joel Benton. 

"A Successful Woman " M. E. W. Sherwood. 

A Blossom from the Hague Wm. E. S. Fales. 

Aims of University Extension Sydney T. Skidmore. 

Polly Patience Stapleton. 

By the Sea Clinton Scollard. 

What Country Girls Can Do Grace H. Dodge. 

Latent Force John Worrell Keely. 

The Personality of the Prince of Wales. Frank A. Burr. 

The Mou jik Julien Gordon. 

Some Letters to Julien Gordon 

John Dickinson Anne H. Wharton. 

Literary Dynamics Francis H. Williams. 

" Maidens Choosing "..'..- Frederic M. Bird. 

With the Wits (Illustrated by leading 
artists) . 

THE NORTH AMERICAN REVIEW. 

CONTENTS OF MAY NUMBER. 

The Wiman Conspiracy Unmasked .... Sir Charles Tupper. 

Canada and the United States Marquis of Lome. 

Napoleon's Views of Religion H. A. Taine. 

f William S. Andrews. 
Common Sense on the Excise Question \ 

( Howard Crosby, D. D. 

The Modern Extinction of Genius Julien Gordon. 

Our Business Prospects Henry Clews. 

Lynch Law and Unrestricted Immigra- 
tion Hon. Henry C. Lodge. 

The Politician and the Pharisee Hon. J. S. Clarkson, Ex-Ass't 

Postmaster General. 



THE BACTERIOLOGICAL WORLD. 385 

NOTES AND COMMENTS. 

Can a Poor Girl Go to College *? Alice Hayes. 

Tight Lacing for Monkeys Edward P. Jackson. 

The Closing Door of Quackery Wm. G. Eggleston, M. D. 

( S. G. Pratt. 

A National Chorus •< 

( Theodore Thomas. 

A Catholic on the School Question Prof. M. F. Egan. 

THE POPULAR SCIENCE MONTHLY. 

CONTENTS OF JUNE NUMBER. 

New Chapters in the Warfare of Sci- 
ence. XII. Miracles and Medicine . 

Part II Andrew D. White, LL. D., 

L. H. D. 

Our Grandfathers Died too Young. . . .Mrs. H. M. Plunkett. 

The Development of American Indus- 
tries since Columbus. V. The 
Manufacture of Wool S. N. Dexter North. (Illus.) 

Questions concerning the Minor Planets. M. F. F. Tisserand. 

The Natchez Indians Howard A. Giddings. 

Survivals from Marriage by Capture ... Lieut. Col. A. B. Ellis. 

The Characteristics of Insects Louis Montillot. (111.) 

" The Pearl of Practice " Elizabeth Robinson. 

Future of the Dry Land M. A. de L' Apparent. 

The Music of the Birds Simeon Pease Cheney. 

Sketch of Nicolaus Copernicus (With Portrait). 

Correspondence : Feet-washing and Feet-kissing. 

Editor's Table : Liberty and Civilization. 

Literary Notices. 

Popular Miscellany. 

Notes. 

THE CENTURY MAGAZINE. 

CONTENTS FOR MAY, 1891. 

Portrait of Emperor Nicholas I. En- 
graved by G. Kruell Frontispiece. 

Game-Fishes of the Florida Reef C. F. Holder. 

Pictures by Victor Perard. 

Illusions Robert Underwood Johnson. 

Salons of the Empire and Restoration. .Amelia Gere Mason. 

With nine portraits. Decorations by A. Brennan. 
The Squirrel Inn. I Frank R. Stockton. 

Pictures by A. B Frost. 

Poetry O. C. Auringer. 

Visible Sound. 

I. Voice-Figures Margaret Watts Hughes. 

Pictures from photographs taken by the author. 



386 THE BACTERIOLOGICAL WORLD. 

II. Comment. 
Four diagrams. 

Of One We Love or Hate Maurice Francis Egan. 

The Faith Doctor. (Begun in February) . Edward Eggleston. 

Ballad of an Old Pine John H. Boner. 

Louisa May Alcott Josephine Lazarus. 

With three portraits. 

A Bulgarian Opera Bouffe F. Hopkinson Smith. 

Pictures by the author. Portraits from photographs by Karastoyanov, Sofia. 

In Disguise Frances Louise Bushnell. 

At the Court of the Czar. In two Parts. 

Part I George Mifflin Dallas. 

Decorations by George Wharton Edwards. 

A Heady Maid Louise Morgan Sill. 

Exhibition of Artists' Scraps and 

Sketches William Lewis Fraser. 

With thirty pictures by the Exhibitors. 

Old Gus Lawson Richard Malcolm Johnston. 

Picture by E. W. Kemble. 
The Confederate Diplomatists and ) 

their Shirt of Nessus. A Chapter - John Bigelow. 

of Secret History ) 

Pioneer Mining in California E. G. Waite. 

Pictures by Harry Fenn, E. W. Kemble, A. C. Redwood, W. L. Dodge, 
A. Castaigne, with suggestions from paintings by Xahl, and from "Punch." 
In Beaver Cove Matt Crim. 

TOPICS OF THE TIME. 

International Copyright Accomplished — Lobby Evils and Remedies. 
The National Conference of Charities and Corrections. 
An American Cheap Money Experiment. 

OPEX LETTERS. 

Certain Criticisms of Certain Tales James Lane Allen. 

The Negro in Nashville C. F. Smith. 

Homeopathy and "Expectant Treat- 
ment " 

" Does Vivisection Help? " Thomas W. Kay, Mary Putnam 

Jacobi, M. D. 

BRIC-A-BRAC. 

Observations 

Friend and Lover M. A. De Vere. 

To a Thermometer W. D. Ellwanger. 

Ashes J. C. Miller. 

When Youth Mounts E. Singleton. 

Love's Young Dream G. D. Litchfield. 

Fame C. H. Crandall. 

The Survival of the Fittest V. F. Boyle. 

The Latest Indian Outrage. Drawn by E. W. Kemble. 



(TL 



D 




A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies. 

Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological World, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to at the 
same office. 

Authors of original articles published in this magazine will be entitled to twenty- 
copies of the number containing it, provided, that they request the same in writing 
when sending the communication. Keprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. - 

Discontinuation :— At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first of next January. 



COLLABORATORS 



Paul Gibier, M. D., New York. 
Prof. W. T. Belfield, IVi. D., Chicago. 
Prof. A. W. McAlester, A. M., M. D., 

Missouri University Med. Dep't. 
Prof. Henry P. Loomis, M. D., 

New York . 



Frof. L. Bremer. M. D., St. Louis, Mo. 
J. W. Stickler, M. D., Orange, N. J. 
Prof. Paul Schweitzer, Ph. D., 

Missouri University. 
Prof. T. J. Burrill, Ph. D., 

Illinois University. 



B.T. 


Gallowat 


, B. S., Section of Vegetable Pathology, Washington, 


D. C. 


Vol, 1, 




COLUMBIA, 


MO, 


U. S. A., JUNE, 1891. 


No. 6. 















EDITORIAL. 



SPECIFICITY OR NON-SPECIFlCITY OF PULMONARY CON- 
SUMPTION. 

In a paper read the third of June before the Pennsylvania 
State Medical Society, Dr. Thos. J. Mays, takes the ground 
that pulmonary consumption is not a specific disease and not 
produced by contagion. The following words of his explain 
the views he holds : 

" It goes without saying that the successful treatment of 
pulmonary consumption depends entirely on a correct and 
rational view of its origin. On this ground, there can be no 
division of opinion. Unfortunately, however, one part of the 
profession holds that the tubercle bacillus is the sole cause of 
the disease ; while the other part believes that the disease 



388 THE BACTERIOLOGICAL WORLD. 

depends on numerous other causes, and that the bacillus is 
merely one of its products or concomitants. 

"Now, it must be confessed that diametrically opposite 
opinions on a question of such tremendous practical and public 
importance are no great credit to a profession which is justly 
' proud of her logical conquests, and which rightfully rejoices in 
her scientific achievements, and in her work of philanthropy. 
Especially is this a misfortune when we consider that the whole 
difficulty rests on an erroneous interpretation of the funda- 
mental terms which are involved in the question ; and, in 
endeavoring to prove this, I desire it to be perfectly understood 
at the outset, that I yield to no one in my admiration of, and 
I might even say in my veneration for, those pains- taking 
explorers who so patiently and assiduously devote their life and 
energies to the investigation of this fearful malady. But I do 
not regard this as a question of personal reverence. It is one 
which must determine the true relation of the consumptive to 
his family, to his friends, and to society ; whether or not he is 
to be looked upon as a center of infection, a source of danger, 
and liable to spread the disease among those who are well. 

"Where then lies the fundamental misapprehension? It 
is, first, in affirming that, because a disease is capable of being 
communicated from one animal to another through inocula- 
tion, that, therefore, it must be contagious or infectious among 
the human species ; second, in holding that the disease ( tuber- 
culosis ) which is produced artificially in animals through inoc- 
ulation is the same as that ( consumption ) which is naturally 
produced in man." 

We believe Prof. Mays perfectly sincere in his opiniou 
and doubt not in the least the noble animus of his excellent 
contribution. 

However, we fail to see in the same light and believe that 
no statistic, bedside or hospital record warrants the assertion nor 
even the assumption that tuberculosis is not a specific affection. 
In fact, the very statistics and reports which the learned Doctor 
quotes copiously are insufficient, in our humble judgment, to 
support his conclusions, and many may be satisfactorily and 
rationally explained under the principle of contagiousness. 

In the first place 1 it is true that a part of the profession 
holds that the bacillus of Koch is the sole cause of the disease 



THE BACTERIOLOGICAL WORLD. 389 

in question whilst the other part holds that the bacillus is " one 
of the products or concomitants of the affection." But if we 
judge from writings, medical journals, discussions, congresses, 
association meetings in the whole universe, the reports of foreign 
academies and learned societies, the oft-repeated opinions of the 
most learned and most practical physicians of all countries, the 
part of the medical profession which holds that the bacillus is 
"one of the products of consumption " sinks into insignificance 
compared with the other part, the mass of practitioners who 
believe the reverse. 

The Doctor gives in the above lines what he holds as 
the fundamental misapprehension that leads so many to believe 
in the contagiousness of tuberculosis. First, he considers it an 
error to conclude that "because a disease is capable of being 
communicated from one animal to another through inoculation, 
that, therefore, it must be contagious or infectious among the 
human species." 

This is undoubtedly a positive evidence that (lower) ani- 
mal tuberculosis is transmissible, and a transmissible disease is 
a specific one. And that tuberculosis of man and beast is 
identical seems positively ■ proven by numerous observations of 
some of the most accurate practitioners as well as laboratory 
investigators, by the transmission of the disease from man to 
animals, not only by inoculation but also by inhalation of 
human tuberculous sputa, thereby causing pulmonary tubercu- 
losis to develop in animals, and also by the ingestion of human 
tuberculous sputa producing tuberculosis in the intestines, etc., 
of the animal subjects so treated. 

The disease thus produced is similar in its action upon the 
organs affected as it is in man when it occurs accidentally. The 
same organisms, the same pathological lesions and the tuber- 
culosis products have the same virulence. 

These facts, which answer the second so-called misappre- 
hension as well as the first of Dr. Mays, are not derived only 
from inoculations among lower animals as you see but by 
transmission of the disease from man to animals by various 
means — inoculation being only one of the various methods of 
conveying it. 

But still more has been done to prove the specificity of 
tuberculosis in man. The Gazette Medical de Paris, 1872, No. 



390 THE BACTERIOLOGICAL WORLD. 

17, gives an account of one inoculation made with sputa on a 
man of 55 years by three bold Greek physicians, Demet, Paras- 
keva and Zallonis, of Syria. 

They had a patient affected with gangrene of the big toe of 
the left foot, and in whom nothing whatever indicated the 
slightest predisposition to tuberculosis. His lungs were sound 
so far as the ear and various methods of diagnosis could find. 
Three weeks after, the experimentalists detected at the summit 
of the right lung the beginning of an induration. The thirty- 
eighth day the patient died from the effects of the gangrene. 
Post-mortem examination revealed seventeen tubercles at 
the summit of the right lung where they had diagnosed indu- 
ration ; two were found on the upper portion of the left lung 
and two more on the liver. The doctors thought the tubercles 
advanced in proportion to the lapse of time between inoculation 
and death. 

Certainly this fact in itself is not conclusive, but it is a 
very strong addition to the many other evidences of the posi- 
tive contagiousness of tuberculosis. 

In denying that by inoculation or transmission we do 
establish the proof of the specific nature of tuberculosis, one 
must also deny that inoculation or any other processes of 
transmission of any other malady is in any sense evidence of 
specificity. 

If this is true, then, the inoculation of glanders between 
animals or from animal to man ; the inoculation of anthrax 
between animals or from animal to man ; the inoculation of 
vaccinia from bovines to man or among the human species ; the 
inoculation of septicemia between animals or from animal to 
man are in no sense evidence of specificity. 

In fact the basis from which Dr. Mays reached his con- 
clusion that the mass of medical men who believe in the con- 
tagiousness of consumption have misinterpreted the bearing of 
inoculation, etc., etc., furnishes equal grounds for the denial of 
the specificity of every known and contagious disease in the 

world,. 

It is only by transmission of a disease that one can posi- 
tively and undeniably establish its specific nature. 



THE BACTERIOLOGICAL WORLD. 391 



LESSONS IN BACTERIOLOGY. 



LESSON FIFTH {continued). 



BY PAUL PAQUIN, M. D. 



CULTIVATION" OF BACTERIA. 

Before proceeding with the second part of this lesson, I 
wish to make clearer, correct the sense, and add to two or three 
statements made in the last lesson. 

First, regarding the sterilization of blood serum, which we 
will see in detail, I wish to say that it may be effected also in a 
water bath though not so safely as in dry air. ( See twelfth 
line from the bottom, page 314, May number of this magazine.) 

Second, a beef broth culture containing gelatine will be 
better and more solid when heated from five to six or ten min- 
utes two to four consecutive days at a moist temperature of not 
over 212° F. ( See page 314, first and second lines from the 
bottom, and the first line at the top of page 315.) 



PAET II. 

ESSENTIAL APPARATUS TO CULTIVATE BACTERIA. 

The first requisite is apparatus of culture presenting sim- 
plicity, solidity, convenience and cheapness as far as possible. 
The second is the proper preparation of culture media. 

Apparatus. — The most commonly used recipients are test 
tubes, the most suitable kind being the narrow, thick tubes 
without flange at the mouth. Little culture flasks, as the 
Pasteur flask for instance, are also in use. However, in a case 
of necessity, one might utilize ordinary little round medicine 
bottles. 



* Illustrations from Salmonsen's Technology, French edition. 



392 THE BACTERIOLOGICAL WORLD. 

The tubes, bottles or flasks are first to be washed carefully 
and thoroughly in distilled or at least boiled water and allowed 
to drain well. A good-sized bundle of cotton wool is twisted 
with the fingers and adjusted exactly, an inch or two in depth, 
in the opening of each, leaving a projection over the edges. 
( See Fig. 7.) This plug must be dense and tight enough to 
prevent the introduction of air germs, but yet not so tight- 
fitting as to be difficult of moving by a gentle, firm tortion 
motion. When plugged they are placed in a wire basket, say, 
or a perforated metallic bucket of small size and the whole 
introduced into a dry heat sterilizer. ( See Fig. 6, A.) 

Leave them there at about 280° F. for ten or twenty min- 
utes or until the cotton wool plugs begin to be slightly brownish. 
Thus sterilized, and as sd*on as cooled, the tubes or bottles are 
ready to serve as recipients of culture media. These apparatus 
are for liquids or semi-liquids. 

There are other culture apparatus in use for such culture 
media, but 1 intend to giYe you only the simplest and most 
economical that may be used advantageously without any 
expensive laboratory arrangement and without the knowledge 
of difficult special details of laboratory technique. For cultiva" 
tion of bacteria on more solid matter, as on potato for instance, 
glass dishes are used as recipients. They are composed of two 
flat glass dishes, the larger serving as cover for the smaller. 
( Cut will appear in next issue.) 

To prepare them they should be well washed as the test 
tubes, drained, and a circular sheet of absorbent paper damp- 
ened with a bi-chloride of mercury solution placed in the bot- 
tom of the smaller dish. The solid culture media are carefully 
placed inside and covered. 

Practice alone will teach the skill necessary for these 
manipulations without the introduction of air germs ; further 
we will note these manipulations more in detail. 

All these apparatus are for the cultivation of aerobic bacteria. 
The anaerobic need special treatment, and we will consider them 
apart. 

There are special apparatus, some sort of incubators, to 
keep the cultures at any temperature desired. Of these we 
shall speak in due time. 




FIG. 6. 



A. Dry Heat Sterilizing Oven; a common metallic box 
covered with pelt. 

B. Koch's Steam Sterilizer: a, hook to suspend objects; 
6, metallic bucket containing plugged test-tubes; 

c, metallic cylinder to augment height of apparatus; 

d, c?, perforated metallic plate; e, water guage. 



CL 



A. 




B. 




FIG. 7. 

A.-a. Chamberland Porcelain Filter bougie; 6, the Filter 'in operation 

by means of Aspirator. 
B. Various Culture Vessels— Test-Tubes and Flasks : I, II, VII plugged 

with cotton— wool; V, VI, provided with tubular corks or plugs. 



THE BACTERIOLOGICAL WORLD. 395 

LESSON SIXTH. 

CULTURE MEDIA AND PROCESSES OF PREPARATION", 

Among the many nutritious preparations to cultivate bac- 
teria, I will here give you only those which have proven suc- 
cessful with the majority of organisms. The laboratory worker 
has his media of predilection and changes them to suit his 
special needs in his particular operations. 

Broths in general. — The juice obtained in boiling lean 
meat, such . as beef, pork, chicken, etc., even with its acid 
reaction is suitable to grow various micro-organisms. But if 
neutralized by the addition of bi-carbonate of soda, it is still 
better and affords proper nourishment for a number of patho- 
genic germs. 

It should be noted, at the onset, that variety in the quantity 
and quality of nutritive principles in a given media may result 
in differences in the growths in general, and even in a measure 
produce different microscopic appearances in the same indi- 
vidual. 

Beep broth. — Take one pound of very lean beef chopped 
fine ; one litre of water, put into an enameled kettle, or a large 
glass flask which may be placed in a water bath, boil thirty 
minutes and filter. The filtrate should then be neutralized by 
adding drop by drop while stirring a saturated solution of 
bi- carbonate or phosphate of soda. Test the liquid often with 
litmus paper while doing this, in order to stop the addition of 
alkaline liquid the moment it becomes neutral or slightly 
alkaline. Add one-half dram of common salt. After this 
boil the filtrate again to coagulate all the insoluble albu- 
menoids. The addition of the white of an egg before beginning 
this second cooking is of material assistance in clarifying the 
broth. Let the liquid stand until cool, filter again, and put 
into the sterilized test tubes, bottles or culture flasks spoken 
of, or put into a large sterilized glass flask to keep as stock 
culture broth to be drawn from at convenience in the future. 
The test tubes or other bottles ( in which the cultures are to be 
made ) should be about one quarter full Eeplace the cotton 
plugs immediately after liquid is in them. 



396 THE BACTERIOLOGICAL WORLD. 

These culture recipients and their broth are then placed in 
a steam sterilizer and heated to over 300°F. five or ten minutes, 
and complete sterilization is effected. In case one has no steam 
sterilizer, the media should be placed in a wire basket or per- 
forated tin pail say, and boiled for about thirty minutes once a 
day for three to six days in a water bath. 

Cloudiness may spoil the appearance and even utility of 
broths thus prepared. This may be avoided by repeating the 
cooking process once or twice and the addition of more egg 
albumen after neutralization as before stated, before putting 
into the test tubes or other culture vessels. 

Five per cent, of pure glycerine added to this before neu- 
tralization renders it proper to grow the bacillus of tubercu-. 
losis. The addition of one to two per cent, of peptone, a little 
cane sugar, etc., is also occasionally resorted to particularly for 
the cultivation of anaerobic organisms. 

Liver broth. — We use in our laboratory, liver broth (sub- 
ject to solidification by gelatine, etc.) which at first I had pre- 
pared somewhat on the principle of beef broth but which was 
afterwards beneficially modified by my assistant, Dr. Paul 
Evans. The modified method is as follows : 

Take one pound of finely chopped beef liver and put in a 
two litre flask with one litre of distilled water. Heat in water 
bath for half an hour at about or below 212° F. and press the 
liquid through a fine gauze or clean linen cloth. Boil the 
liquid for an hour or more and filter. Put in four per cent, 
glycerine, carefully neutralize with sodium carbonate (make 
slightly alkaline rather than acid). Add white of an egg and 
boil again a few minutes. Filter again. Sterilize carefully one 
hour once a day for three to four days at boiling point in steam 
sterilizer or water bath. 

Another fluid which I have used in the laboratory is the 
following : 

Phosphate of magnesium 20 grains 

Chloride of sodium . . . . 20 - " 

Carb. of sodium. .... 15 " 

Nitrate of ammonium 10 " 

Carbonate of potassium 10 " 

Sulphuric acid ........ 30 drops 

Water 6 ounces 

Beef broth (prepared as stated) 1 litre 



THE BACTERIOLOGICAL WORLD. . 397 

Dissolve the salts in a part of the water, say half, and the 
acid in the remainder, then add each solution to the broth 
while stirring. Neutralize, put in culture tubes or flasks and 
sterilize at 212° F. repeatedly, or at once at over 300° F. 

This is a special medium used in special investigations as 
we may see further, but not proper for the growth of all organ- 
isms. 

Vegetable infusioxs.— Besides these animal substances, 
various vegetable infusions are used as nutritive media. 
Thus hay, wheat, cabbage, potato infusions made by boiling 
these and sterilization as for other substances are occasionally 
employed. I need not dwell on these. They are very simple. 

Again decoctions of fruits are in use for the growth of 
organisms such as fungi, moulds, etc. Among these may be 
mentioned the plum decoction that Salmonsen speaks of. I 
have cultivated organisms of various kinds on fresh ripe grape 
decoctions. 

In the study of pathogenic bacteria acting on animal tis- 
sues these are of little value except the potato decoction which 
has some utility. 

Some solid cultures. — The animal broths described may 
be solidified very profitably for the cultivation of bacteria on 
a smooth surface or in the center of their transparent sub- 
stance, thus affording the opportunity of following the growths 
step by step with the naked eye, watching their progress, 
appearance and action on the nutrient matter. 

To solidify the meat broths, gelatine or agar-agar (from 
gelatinous algues) or both are added as follows : Add from 5 to 
10 per cent, of the best quality of gelatine in clear thin plates or 
fibers to the broth before neutralization. Dissolve at a low heat, 
neutralize or make slightly alkaline, then heat to about boil- 
ing point ; filter whilst hot for in cooling or after cooling, it will 
not filter. There are water baths, filters or hot water filters 
on the market for this purpose. They have the advantage of 
keeping the liquid at a constant heat whilst being filtered 
directly into a sterilized stock flask or into the culture tubes. 

In using agar-agar, the broth should be made slightly alkaline 
before its addition because if cooked in an acid solution it will 



398 THE BACTERIOLOGICAL WORLD. 

form sugar. The remainder of the process of preparation is as 
for gelatine. The quantity used is 1-5 to 1-2 per cent. 

Jensen had the happy idea to combine agar-agar and gela- 
tine and thus improve the jelly medium. This combination is 
effected in the proportion of 5 per cent, of gelatine and 0.75 
per cent, of agar, taking care to add the agar, as just stated 
after a slight alkalinization, or at least neutralization of the 
fluid, and not before as is done with gelatine. 

(Sixth lesson continued in July number.) 



ORIGINAL ARTICLES. 



SORE THROAT AND PURULENT PLEURISY WITH STREP- 
TOCOCCI. 



BY M. HANOT, 

Before the Hospital Medical Society, Paris. 

We admit to-day that a certain number of sore throats are 
microbic in origin, but the precise documents on the matter do 
not permit us to establish yet a new satisfactory classification. 
This is why I have thought it my duty to present to the society 
the observation of a sore throat by streptococci, the evolution 
of which has shown that such a sore throat may present signs 
to the deepest infection and be followed with the gravest com- 
plications. 

Here is, in short, this observation : 

X , aged 19 years, resident of Paris for 5 months, came 

under my care the seventh of March, 1889 ; nothing in the 
antecedents. 

Twelve days previous there was a violent sore throat, with 
lassitude, anorexy, fever, prostration. In the night of the third 
to fourth, violent stitch in the side. 

At his entrance into the hospital the prostration was great. 
The patient responded painfully to the questions we addressed 



THE BACTERIOLOGICAL WORLD. 399 

to him. Feverish tongue. Uniform red tint of the pharyngeal 
mucous surfaces. The tonsils did not seem augmented in vol- 
ume, but they were painful. No false membranes ; not of 
adenopathy, nor epistaxis, nor vomiting ; liver extending the 
false ribs a finger's breadth ; urine slightly urobilic, with a 
cloud of albumen ; acute dyspepsia ; signs of pleuritic effusion 
in the right, in the inferior, two-thirds ; pulse, 90 ; temperature 
in the neighborhood of 100^° F. 

The general state is worse than could be expected in the 
ordinary pleurisy. Therefore we diagnosed an infectious pleurisy 
of pharyngeal origin. 

Treatment : milk, broth, tonics, 15 grains of sulphate of 
quinine. 

The eleventh of March the prostration is the same, diarrhoea 
augmented. An exploring puncture gives issue to a little 
clouded, flaky liquid, in which we find numerous streptococci. 
An evacuating opening brings forth afterwards nearly a pint of 
liquid. 

The twenty-first, the general state and local condition being 
the same, a new puncture permits the evacuation of one pound 
of purulent liquid. Temperature over 102° F. ; pulse, 128 ; 
respiration, 50. 

The twenty-fifth, empyema in place of election ; issue of a 
litre of pus ; antiseptic wash of the pleural cavity ( solution of 
chloride of zinc 2 per cent., and boric solution 4 per cent); 
drainage. 

The temperature, which in the morning was nearly 103° F., 
instead of falling, as in the fortunate cases, rose to above 104°F.' 
The general state also remains serious. The patient is as if mum- 
mified in his bed, and presents the maximum signs of infection. 
The muscles are literally melted ; dull color ; complete anorexia. 
Finally, death, the twenty-fourth of April. 

At the autopsy we ascertained that the amygdales, hardly 
more voluminous than normal, contained small abscesses of the 
size of the head of a pin to that of a pea. The tissue, retro- 
pharyngeal, cellular tissue, is the seat of a well-confined lateral 
purulent streak, a centimetre and a half in width, directing 
itself towards the right, from the summit of the thoracic cage. 
There, loosening the parietal pleura of the thoracic wall, it 



400 THE BACTERIOLOGICAL WORLD. 

descended to the level of the inferior third of the chest, where 
it opened in the interior of the serous cavity. The right pleural 
cavity did not contain liquid ; its walls were covered with thin 
grayish false membranes. The right lung is congested and not 
hepatized in the. superior two-thirds, atelectasis in its inferior 
third. No tubercles. The left lung was wholly congested. 

The purulent channel contained some streptococci. The 
same micro-organisms have been found in the amygdalien 
abscesses. 

Nothing particular to note in the other organs. 

I will draw your attention in connection with this case 
that Messrs. Oornil and Babes, in their book, mentioned the 
presence of streptococci in abscesses of the amygdales. 

There was described, said Mr. Fernet, in discussing the 
above report, a certain [number of serous pleurisies ; these are 
tuberculous, rheumatic, scarlatinous, pleuritic, consecutive to 
other diseases, or to^the affection of the neighboring organs. 

Mr. Netter has said to us that, bacteriologically, 70 per 
cent, of serous pleurisies were tuberculous. And in the 30 per 
cent, which remained Mr. Talamon has found twice the pneu- 
mococcus, three times the streptococcus, in 17 cases of pleurisy 
that he has examined ; we are able then to admit that certain 
cases of serous pleurisy are produced by the pneumococcus and 
by the streptococcus. I have just observed one in which Mr. 
■Grirode has found the Eberth bacillus. We have noticed some- 
times pleurisy coming unexpectedly as complications of typhoid 
fever. 

Messrs. Eendu and Yalentini have rejDorted each an obser- 
vation of purulent pleurisy ; recently, Mr. Charrin reported an 
observation of hemorrhagic pleurisy. No one has yet reported 
a case of serofibrinous pleurisy in which has been found the 
Eberth bacillus. 

The case was that of a young man aged 18, who pre- 
sented all the symptoms of typhoid fever ; the rosy lenticular 
spots were lacking, it is true, but at the fifteenth day of the 
affection this fact was nothing astonishing, and this was on the 
fifteenth day. In searching well in the antecedents, however, 
we found that the affection had begun six weeks before ; the 
disease had at this epoch a stitch in the side and symptoms 



THE BACTERIOLOGICAL WORLD. 401 

which permit the supposition that the effusion dated back to 
that time. And it is at the moment that convalescence seemed 
to establish itself that symptoms analogous to those of the 
typhoid fever supervene. A complete examination at the time 
of his entrance to the hospital led to the discovery of a serous 
pleurisy in which Mr. G-irode found the Eberth bacillus. 

I have observed several analogous cases from a symptomol- 
ogical point of view, though bacteriological examinations were 
either omitted or remained negative, and I ask myself if there 
is not reason to admit a variety of serofibrinous pleurisy of a 
typhoidic character. 

Mr. Ohantemesse remarked, in the discussion, that there 
are perhaps in certain cases some infections by typhic bacilli. 
It is thus, he said, that I have seen a case of typhoid fever last 
five months; that, in another patient, I have seen abscesses due 
to typhic bacilli occur unexpectedly fifteen months after the 
typhoid fever ; that, finally, in a third, I have seen an osteoar- 
thritis which had lasted three years and which had originated 
six weeks after a typhoid fever. These facts show how long may 
last the virulence oi the typhic bacillus. Mr. Dupre has found 
in the pus of a biliary vesicle the Eberth bacillus still virulent 
a very long time after a typhoid fever. — Report from La 
Tribune Medicate . 



402 THE BACTERIOLOGICAL WORLD. 



MEDICAL BACTERIOLOGY. 



THE ELIMINATIVE TREATMENT OF PUERPURAL SEPTI- 
CEMIA. 



From a study of infectious diseases in general, and from 
observations on the methods of elimination of toxic substances 
from the organism by the natural emunctories, Heliodor de 
Swiecicki ( Gazette Hebdomadaire de Medecine et de Chirurgie) 
advises the employment of these means to rid the system of 
poison in puerperal septicemia. Of course, the early treatment 
of this disease, if the case is seen in time, is, first, by asepis and 
and antisepsis by means of uterine irrigation, etc.; and, sec- 
ondly, by the use of alcohol and every means to prevent the 
poison from overpowering the organism. But, if the morbid 
germs and their products have surcharged the blood, the author 
says, the production of exaggerated diuresis and diaphoresis 
would be a rational method of eliminating the morbid elements. 
The treatment consists of subcutaneous injections of pilocarpine 
and the drinking of large and frequent draughts of sterilized 
chloride of sodium solution. If there is any difficulty in the 
patient's swallowing the solution, it is to be introduced directly 
into the stomach by means of a tube. The author reports the 
success of this method of treatment in several grave cases of 
puerperal septicaemia. The amelioration of the symptoms was 
prompt, the temperature falling, the pulse becoming less rapid, 
and the rigors disappearing in a very short time. The patient 
must be watched closely for any sign of heart failure; in the 
event of which small doses of alcohol are to be given. — 
Exchange. 



NEW TREATMENT OF PHTHISIS. 



BY GERMAIN SEE. 

The author shuts his patient up for two, three or more 
hours daily in a hermetically closed metallic chamber, into which 
is slowly admitted a current of compressed air, which, having 



THE BACTERIOLOGICAL WORLD. 403 

passed through a mixture of creosote and eucalyptol, is saturated 
with the vapor of these substances. Since August last ten cases 
of phthisis have been submitted to this treatment, all of which 
cases, with one exception, had reached the period of soften- 
ing, and bacilli had been detected in the sputa. The results 
obtained were : Return of appetite, even in advanced cases, gain 
of weight and strength, fall of temperature to the normal in a 
week or two, disappearance of haemoptysis, diminution of cough 
and of purulency of sputa, cessation of dyspnoea. It is claimed 
that the method reduces the malady to a purely local lesion, all 
the general symptoms disappearing, even though rales may per- 
sist. M. See related the history of seven of his cases, all of 
which were relieved, and some actually cured. The treatment 
has been found efficacious in fetid bronchitis (dilatation of the 
bronchi ). 



SURGICAL BACTERIOLOGY. 



THE ACTION OF TRYPSIN, PANCREATIC EXTRACT AND 
PEPSIN UPON S TOUGHS, COAGULA AND MUCO-PUS. 



BYE. T. MORRIS. 



Something more than peroxide of hydrogen is wanted for 
managing the abscess cavities of tuberculous bone disease to our 
entire satisfaction. It is true that with hydrogen peroxide at 
hand we can open the biggest of cold abscesses m a feeling of 
security for the patient, and yet it is desirable to get the tough 
lining membrane of fibrinous lymph and the loose coagula and 
the sloughing diseased tissues out of the way as thoroughly as pos- 
sible ; for once these substances have begun to seethe in fermen- 
tative processes, we are hard pressed to keep the patient out of 
trouble. 

The sharp spoon and scissors will do the work, to be sure, 
but only after tedious labor and egregious wounding of the 
weak patient. 

A resource was brought into play a few weeks ago when I 
had occasion to make suggestions relative to the treatment of a 



404 THE BACTERIOLOGICAL WORLD. 

crushed liver. Portions of the organ, which were dark and 
sloughing, remained so firmly attached that their removal was 
dangerous, and the pultaceous lining membrane of the enor- 
mous abscess seemed to invite all manner of microbe guests. 
The idea of liquefying the dead tissues with a digestive ferment 
came into mind, and this being suggested was carried into 
effect by the family physician, who injected into the abscess 
cavity a solution of scale pepsin, and writing to me afterward, 
said : "The pepsin did mighty good work. It broke up all 
dead tissues, rendering them mostly liquid, and changed the 
color from brown to straw color. The liquefied substances were 
easily washed out through the drainage-tube. The wound was 
sterilized daily afterward with hydrogen peroxide, and the 
patient recovered without a bad symptom." 

On theoretical grounds I had supposed that trypsin would 
be the best liquefier, that pancreatic extract would stand next 
in value, and that pepsin would be used only when it was 
inconvenient to obtain the other ferments ; but when it came 
to a practical test, pepsin led all the rest. Experiments, which 
were carefully made for me in detail by Dr. Charles N". Haskell, 
I will refer to only in a brief way here. Dr. Haskell used 
tough, partially dried blood coagula for test purposes, and the 
trypsin, pancreatic extract and pepsin were fresh. As a result 
of numerous experiments, it was determined that four grams 
of pepsin dissolved in three hundred cubic centimetres water, 
acidulated with one per cent, of hydrochloric acid and applied 
to one hundred grams of the coagula at a temperature some- 
what above 100° F., would liquefy the coagula in thirty-six 
minutes. 

Pancreatic extract used in alkaline' solution, other factors 
being as in the pepsin experiment, required two hours and 
twenty-six minutes for liquefying the coagula, and at the end 
of that time little tough knots of fibrin still remained. 

Trypsin in alkaline solution, and used in the same propor- 
tions and under the same conditions as the pancreatic extract 
and the pepsin, required two hours and ten minutes for lique- 
fying the one hundred grams of coagula. 



THE BACTEEIOLOGICAL WORLD. 405 

From these experiments it seems that a 10-per-cent. solu- 
tion of the best pepsin acidulated with 1 per cent, of hydro- 
chloric acid, and heated to a temperature above 100° F. ( not 
over 120° F. ), will be proper for surgical purposes. 

We need not employ the pepsin until the patient has recov- 
ered from the effects of ether after an operation, and then the 
liquefying process can be attended to at leisure. The abscess 
cavity should be washed out with boiled water, for antiseptic 
solutions would interfere with the action of a digestive ferment. 
The patient then assuming a good position for holding the 
pepsin solution in the abscess cavity, he can receive the hot 
injection ; and hot fomentations continued for an hour will 
promote the action of the ferment down below. 

Bad tissues sufficiently liquefied are washed out with boiled 
water, and the whole wound is then sterilized with peroxide of 
hydrogen and prepared according to the surgical conception of 
neatness. 

The facts relative to the comparative activity of the diges- 
tive ferments were not known to me at the time when I asked 
friends to collect practical points for this paper. 

Dr. 0. N". Haskell liquefied two grams of tough lining 
membrane from the tuberculous abscess of a case of hip-joint 
disease with pepsin in fifty-five minutes. 

Dr. C. D. Jones, of Brooklyn, poured a solution of pancre- 
atic extract (pancreatic extract, 5^j j water, gviij ) into the 
abscess cavity of a case of hip-joint disease one week after the 
operation of excision had been perfected. He then wrote me 
as follows : ' * The solution was allowed to remain in place half 
an hour, and the result was remarkable. Upon irrigation, I 
washed out numerous shreds of broken-down ligamentous tissue 
and many spicula of dead bone that had become imbedded in 
the soft tissues and that had previously escaped both irrigator 
and curette. The wound was then flushed out with hydrogen 
peroxide, and this treatment was followed by a marked improve- 
•ment in the patient's general condition." 

Dr. L. A. Zerega applied a thick solution of pancreatic 
extract to the large slough of a burned wound of the chest, but 
four hours were required for breaking up and separation of the 



406 THE BACTERIOLOGICAL WORLD. 

slough. A thick solution of the pancreatic extract applied to 
the large sloughs of a case of fracture of the spine failed to 
liquefy the sloughs after prolonged application. These two 
experiments were made at my suggestion, and were wrong in 
theory, as later facts have shown. If a thin solution of pepsin 
could be applied to such sloughs in a proper way, they would 
succumb pretty rapidly ; but it is only in case of sloughs situ- 
ated deep in cavities that we can apply the pepsin in a practical 
way. 

In one case in which the bladder contained blood- clots and 
the catarrhal mucous membrane discharged ropy muco-pus, 
pepsin injected for the purpose of liquefying the clots not only 
fulfilled its mission in that direction, but unexpectedly cleared 
out the muco-pus and left the interior of the bladder quite 
clean. The process was repeated as soon as the muco-pus again 
became abundant, and the patient experienced a feeling of 
relief after the simple cleansing that pepsin afforded. 

I much regret that pepsin was not used in one of my cases 
of empyema following a bullet wound of the chest. The chest 
cavity was walled off into compartments by fibrinous bands, and 
these compartments contained large lymph coagula which were 
removed with great difficulty. Pepsin would have made simple 
work of breaking down the bands and liquefying the lymph 
coagula. 

It is not easy to see at a glance the whole field for digestive 
ferments in surgery, but we know that they are bland and 
harmless in any proportion, and that they will liquefy dead 
tissues close down to living ones, and that their action will 
then end abruptly. The harmless employment of digestive fer- 
ments should be the means of saving many a life that otherwise 
would flicker and go out under the waves of septic infection 
from decomposing organic matter in the depths of tuberculous 
abscesses. — Xeiu York Medical Journal. 



THE BACTERIOLOGICAL WORLD. 407 



SELECTIONS. 



STUDIES ON THE ACTION OF DEAD BACTERIA IN THE 

LIVING BODY. 



BY T. MITCHELL PRUDDEN, M, D. 

Director of the Laboratory of the Alumni Association, College of Phy- 
sicians and Surgeons. New York. 

First Article. — Introductory. 

The researches of the past decade, bringing to light, one 
after another, the specific micro-organisms of some of the most 
common and fatal diseases, have been so surprising, so definite, 
so full of the promise of fruitful outlooks upon hitherto untrod- 
den fields, that we have scarcely yet had time to recover from 
the glamour of the new light or to realize, in the urgency of 
fresh practical problems, the exact extent and bearings of the 
new knowledge. 

For a time it seemed enough, and even more than enough, 
that month by month the proof grew stronger that anthrax, 
tuberculosis, typhoid fever, erysipelas, pneumonia, tetanus, 
diphtheria, and a whole group of allied "wound diseases" and 
others of the so-called "infectious" class, were always associ- 
ated with certain germs, each peculiar in its life history, and 
each standing, as we say, in an etiological relationship to its 
particular disease. 

But as we have little by little become accustomed to the 
new light, it has become evident not only that we are still 
ignorant about the relationship of micro-organisms to several 
extremely frequent and important infectious diseases — the 
exanthemata, for example — but also that when we have learned 
that a given acute infectious disease is always associated with a 
particular form of germ, when the life history of that germ is 
made out, and we can say that it stands in an etiological rela- 
tionship to the disease, there yet remains a series of accessory 
problems to be solved in each particular case scarcely less 



408 THE BACTEEIOLOGICAL WORLD. 

important than the establishment of the invariable association 
of the germ with the disease. 

We are just beginning fairly to realize that the disease 
is not an entity, a thing imparted by the invading germ to 
the body, but that it is the result of the reaction of the 
body cells in the presence of the germs ; that the body-cell 
•factor is just as important and just as much in need of study 
as is the germ-cell factor. We have been largely forgetful, 
hitherto, as with painful detail the characters and preferences 
and metabolisms and vulnerabilities of the pathogenic germs 
have one by one been brought to light, that, before our knowl- 
edge of the acute infectious diseases can be at all complete, the 
characters and preferences and metabolisms and vulnerabilities 
of the body cells must be subjected to an equally careful 
scrutiny. The germ side of the problem is new and fascinating ; 
the man side is old, and cellular pathology is a phrase familiar 
to our ears. But these old problems have become fairly new in 
their new light, and cannot too soon be taken up afresh if our 
knowledge of the acute infectious diseases is to be symmetrical 
and of lasting use. 

Partly by clinical observation and partly by laboratory 
studies is the new knowledge of the man side of this theme to 
be acquired, and old clinical observations, which have lain unin- 
terpreted or misinterpreted, and new facts which the new 
points of view cannot fail to elicit, will surely be fast forth- 
coming. 

But, returning to the bacterial side of the problem, it 
became evident, very soon after the definite status of pathogenic 
germs was made out, that something more than their mere 
presence was necessary to account for the manifestations of the 
acute infectious diseases. 

The early discovery that certain pathogenic germs set free 
poisonous substances of one kind or another as the result of 
their life processes, and the evidence that these substances were 
directly accountable for many of the manifestations of the acute 
infectious diseases, drew attention to the complexity of the 
problems involved, called in the services of the physiological 
chemists, and for a time it seemed, and still does to many, that, 
after all, it was the poisons which the bacteria elaborated and 



THE BACTEKIOLOGICAL WORLD. 409 

sent out into the body on their destructive missions which was 
the most important thing. "Ptomaines" became a favorite 
word. When we had said that a given germ produced a given 
disease or effect by the elaboration of a given ptomaine, it 
seemed to many fairly unreasonable to ask for any further 
explanation of the acute, infectious diseases. The germs were 
relegated to the more humble function of poison-factories, and 
the ptomaines were invested with the insignia of malevolent 
power. The pendulum seems disposed to swing back germward 
now, and in this paper, which is preliminary to the record of 
some experimental studies made by Dr. Hodenpyl and the 
writer, and shortly to follow, on the tubercle bacillus, it is my 
purpose briefly to review a series of recent studies on the germ- 
cell bodies which throw a curiously interesting new light on 
some old body-cell problems. 

A very curious vital phenomenon which has long been 
known in certain unicellular organisms — such as the fresh- 
water amoeba and in the leucocytes of both the cold and warm- 
blooded animals — is their response by movement to contact 
with solid substances. Thus the amoeba floating free in fluids 
tends to assume a spheroidal form and to remain immobile. 
When, however, under suitable conditions, it touches a solid 
surface, like that of a glass slide, it sends out pseudopodia and 
performs those curious progressive evolutions known as the 
amoeboid movement. Essentially, the same series of movements 
is observed in leucocytes when they, under favorable conditions, 
come in contact with solid surfaces— such as a glass slide or the 
walls of the body lymph-spaces. This faculty in these primi- 
tive forms of life, consistiug of a simple lump of protoplasm, is 
called tactile sensibility, and it is in virtue of this that many of 
the remarkable and useful evolutions of the leucocytes in the 
body transpire. 

If was found by Pfeffer ( 1 ), a good while ago, that some 
of the lowly vegetable organisms endowed with locomotion 
— the flagellata, bacteria, etc. — were capable of moving toward 
or away from certain substances which exerted a chemical 
action upon them. This property he designated as cliemotaxis, 
and further postulated as positive cliemotaxis the attracting 



410 THE BACTERIOLOGICAL WORLD. 

effect, and as negative chemotaxis the repelling effect, on such 
organisms of the chemical substances. 

Pfeffer has shown that mobile bacteria move toward nutri- 
ent substances, such as beef-tea, and Engelmann and others (2) 
have demonstrated their movement toward oxygen, both effects 
being apparently due to the positive chemotactic action of these 
substances. Stahl (3) showed that similar properties exist 
in the plasmodia'of myxomycetes. This movement has been 
proved to be due, not to currents in the fluids, not to diffusion, 
but to the specific action of the particular chemical substances 
in question on living organisms. 

The chemotactic powers of the juice of raw potatoes, which 
contains, as Pfeffer showed, potash salts and asparagin, has 
been used in capillary tubes by Ali Cohen ( 4 ) to separate 
mobile from immobile bacteria in mixtures. In this way he 
found that he could separate cholera and typhoid bacilli from 
the numerous other forms in the faeces, and thus make easier 
the obtaining of pure cultures for diagnostic purposes. 

Now, the same condition of affairs exists in the leucocytes 
of both the cold-blooded and warm-blooded animals, and the 
conditions and bearings in them of this positive and negative 
chemotaxis were studied in detail by Massart and Bordet ( 5 ) 
and by Gabritchevski ( 6 ) in 1890. The latter observer has 
grouped as the result of his experiments certain chemical 
substances in accordance with their action in this way upon 
leucocytes. Thus, in the group of substances exciting a nega- 
tive — repelling — chemotaxis, we have concentrated salt solution, 
10 per cent. ; lactic acid ; quinine, 0.5 per cent. ; alcohol, 10 
percent.; chloroform; jequirity ; glycerin; bile. Substances 
having no effect — indifferent chemotaxis — are distilled water ; 
dilute salt solution, 0.1 to 1 per cent. ; carbolic acid, 1-per-cent. 
solution; antipyrme ; glycogen; peptone; beef -tea; blood; 
aqueous humor. Among the most prominent substances excit- 
ing a positive chemotaxis are especially sterilized and non- 
sterilized cultures of various pathogenic and non-pathogenic 
bacteria. 

The general method of testing the powers of these various 
substances is to fill small capillary glass tubes, closed at one 
end, with the substance to be tested, and to thrust these beneath 



THE BACTERIOLOGICAL WORLD. 411 

the skin of an animal. After a few hours these tubes are with- 
drawn and their contents examined. Into tubes filled with 
substances inciting positive chemotaxis the leucocytes crowd 
in great numbers, while they are held away from tubes having 
negative chemotactic contents, and when filled with indifferent 
substances there is no effect at all. While the tactile sensibility 
of leucocytes may cause them to cluster in small numbers about 
the surface of the glass tubes, the effect of this property in the 
leucocytes is altogether insignificant as compared with the 
chemical substance exciting positive chemotaxis. 

It appears, then, that there are certain substances associated 
with bacteria which excite in the leucocytes a movement toward 
the germs. The culture medium itself has no such effect, but 
the action is developed equally whether the cultures be living 
or have been killed by boiling. It would thus appear that 
either the bacteria themselves or some result of their life and 
growth must be the exciting agency. 

Under the dominant views regarding the significance of 
the various chemical substances set free by bacteria as they 
grow, it has been assumed that it was largely under the excit- 
ing influence of the ptomaines that leucocytes exhibited the 
phenomena of chemotaxis in the presence of bacteria. A prac- 
tical bearing was given to the subject, under the influence of 
this view, by the assumption that in the process of suppura- 
tion, as commonly induced by various species of bacteria, the 
leucocytes gathering at the inflammatory foci were drawn 
thither in virtue of their chemotactic properties which the 
metabolic bacterial poisons brought into play. To this view 
the doctrine of phagocytosis, as held by Metschnikoff and his 
adherents, readily attached itself, and we had a well-rounded 
hypothesis, in accordance with which the leucocytes, drawn, in 
virtue of their chemotaxis, into the vicinity of invading bacteria, 
at once set to work to destroy them, and with them the poison 
sources which were stimulating excessive cell inroads. 

But, while these observations were going on, an allied but 
quite independent series of experiments was being carried out by 
Buchner and his associates in Munich, which have thrown a 
new and apparently most significant light upon both the phe- 



412 THE BACTERIOLOGICAL WORLD. 

nomena of chemptaxis and the nature of suppuration. To these 
experiments let us then briefly turn. 

While it is fully established that a true suppurative 
inflammation may be experimentally induced by a variety of 
inorganic substances, it is still true that the suppurative pro- 
cesses which occur in the body, either as independent lesions or 
as complications of a variety of diseases, are practically always 
due to the action of bacteria. So that in a clinical sense the 
summary statement, "no suppuration without bacteria," is 
true. While, as above indicated, it has been the general belief 
of late that the metabolic products of bacterial life, the 
"ptomaines" or the " toxines," were the active agents in 
inducing suppuration, this, save in a few instances, has not 
been proved. 

Buchner ( 7 ), m the course of some experiments on the 
introduction of anthrax spores and anthrax bacilli into the 
trachea of rabbits and guinea-pigs, had observed some time 
ago that while the introduction of the bacilli was followed by 
an intense inflammatory reaction of the lung tissue, with 
accumulation of leucocytes, fibrin, etc., in the air spaces, the 
introduction of the spores alone was followed by no such marked 
inflammatory reaction, but that the spores entered the blood 
channels and induced, in due time, -the usual systemic effects 
of anthrax poisoning. 

There is one factor — so reasoned Buchner — which has not 
been taken definitely into account in the causation of suppura- 
tive inflammation by bacteria, and that is the possibility that 
the effect may be produced, not by the ptomaines, not by the 
toxines already so much studied, but by the albumenoid con- 
stituents of the bacteria cells themselves. If this were true, 
then the intense exudative inflammation in the lungs following 
the introduction of the anthrax bacilli might be explained by 
the local disintegration of the bacilli and the setting free of 
their potent proteid constituents, while no such effect would 
follow the introduction of spores. 

It has been repeatedly shown by numerous observers ( 8 ) 
that sterilized cultures of various pyogenic bacteria — such as 
staphylococcus pyogenes aureus, bacillus pyocyaneus, etc, — 



THE BACTERIOLOGICAL WORLD. 413 

were as capable of producing suppuration as were the fresh liv- 
ing cultures. But it was believed that this was due to the reten- 
tion of a toxic substance furnished by the life processes of 
the germ which had not been destroyed by the sterilization, but 
clung about the dead germ bodies. Although Wyosokowitsch 
had filtered off the fluid from sterilized anthrax cultures and 
found that the filtrate was not pyogenic, while the solid 
material was, he inferred only that the toxic material assumed 
to cause suppuration was not soluble in the nutrient fluid. 

Buchner had also shown ( 9 ), in the course of some other 
experiments, that the sterilized emulsion of the so-called 
pneumo-bacillus of Friedlander, subcutaneously injected, could 
cause suppuration in rabbits and guinea-pigs. He found fur- 
ther that if such a sterilized emulsion were allowed to stand for 
some time, so that the solid could be separated from the fluid 
parts of the mass, the fluid part did not cause suppuration, 
while the solid part did. That the effect of such sterilized 
bacterial emulsions was not due to their mechanical effects in 
the tissues was shown by such control experiments as the intro- 
duction of powdered charcoal, infusorial earth, magnesia, 
potato emulsion, etc., beneath the skin, with negative results. 

By a series of manipulations similar to that practiced 
with the pneumo-bacillus, Buchner now tested the effect of 
sterilized emulsions of cultures of seventeen different species of 
bacteria, among which may be mentioned staphylococcus pyo - 
genes aureus, staphylococcus cereus Jlavus, sarcina aurantiaca, 
bacillus procligiosus, bacillus fitzianus, bacillus cyanogenus, 
bacillus megatherium, bacillus subtilis, bacillus coli communis, 
bacillus acicli lactici, bacillus anthracis, proteus vulgaris, 
Finkler's comma bacillus, etc. The injection of one cubic 
centimetre of the sterilized emulsions of each of these germs 
resulted within two or three days in an aseptic — that is, bacte- 
ria-free — purulent infiltration in the subcutaneous tissue at the 
seat of injection. On the other hand, the clear fluid obtained 
by sedimentation from the sterilized emulsions of bacillus cya- 
nogemis, bacillus megatherium and bacillus anthracis induced 
no suppuration while the separated sediment invariably did. 

While it thus seemed probable that the albuminous mate- 
rial of the bacterial cell was at least chiefly concerned in 



414 THE BACTERIOLOGICAL WORLD. 

inducing suppuration on the injection of sterilized emulsions, 
this was not yet fairly proved. Buchner now sought to 
strengthen the evidence by a very ingenious experiment. The 
modern technique of staining bacteria with the aniline dyes 
depends, as is well known, upon the power of these dyes to 
enter into chemical combination with the bacterial cell plasma. 
Acting upon this principle, Buchner found that if he added to 
a sterilized emulsion of thepneumo-bacillus, which was strongly 
pyogenic, an aqueous solution of methyl violet, the emulsion 
was wholly bereft of its pyogenic powers. Anent of this inter- 
esting bit of evidence of the importance of the bacterial cell 
proteids, Buchner calls attention to its bearing upon the theory 
of the antiseptic and antipyogenic action of the so-called 
pyoktanin of Stilling, the usefulness of which in practice is 
still sub judice. 

But more definite proof of the importance of the bacterio- 
protein in inducing suppuration was still needed and Buchner 
proceeded to separate it from cultures of the pneumo-bacillus 
after the method of K"ecki, by digestion of masses of culture in 
dilute potash and precipitation with acetic or hydrochloric 
acid. The precipitate separated by filtration was again dis- 
solved in dilute potash solution and reprecipitated. This was 
done the third time, and at last the purified product was brought 
into solution. This material gave the chemical reaction of an 
albumenoid body. Subcutaneous injection of this material in 
rabbits in some cases was followed by a gathering of leucocytes, 
in others not. As it seemed likely that on simple subcutaneous 
injection the material was readily and rapidly absorbed before 
it produced local effects, recourse was had to a method of 
experiment used by Councilman in his well-known studies on 
suppuration (10). Small glass tubes, drawn out at the ends, 
were filled with pneumo-bacillus protein, sealed up, and steril- 
ized by steam for an hour. These were then introduced, with 
strict antiseptic precautions, beneath the skin of rabbits, shoved 
away from the opening, and, after they were healed in, their 
tips were broken off. After fi^e days the tubes were exposed. 
Around the openings of these, as well as extending deep into 
their interior, were masses and plugs of leucocytes. Cultures 



THE BACTERIOLOGICAL WORLD. 415 

showed no living bacteria. Control experiments with tubes 
filled with salt solution showed no collection of leucocytes. 

If thus seemed to be proved that the pyogenic action of 
sterilized cultures of Friedlander's pneumo-bacillus is due to 
the freed albumenoid constituents of the bacteria cell. That 
such a freeing of the contents of the bacterial cells occurs in the 
tissues of the living body seems evident from their well-known 
proneness to disintegration and the development of involution 
forms in suppurative foci. 

The next thing to be done was to carry on a similar series 
of experiments with other well-known pathogenic bacteria. To 
this task Buchner and his associates addressed themselves in a 
series of studies as yet not fully published ( 11 ). But, even so 
far as their results are known, some most significant facts have 
been elicited. Buchner endeavored to separate by the method 
of Necki (see above ) the bacterio -protein from about fifteen 
species of bacteria, but in many of these the attempt was 
unsuccessful, because sufficient solution and extraction of the 
proteid ingredients of the germs did not occur. The bacillus 
pyocyaneus gave the most abundant albuminous extract, but a 
sufficient amount was obtained from staphylococcus pyogenes 
aureus, bacillus typhosus, bacillus subtilis, bacillus acidi lactici, 
and from the red potato bacillus for animal experiment. It 
was, in fact, found that capillary tubes filled with the purified 
proteids from all these species of bacteria and placed beneath 
the skin of the rabbit showed after two or three days, extend- 
ing into the open end, a plug of fibrinous pus several millime- 
tres in length. This plug was found, on microscopical 
examination, to consist largely of leucocytes. 

That the ordinary chemical decomposition products of 
bacterial cell life are not concerned in inducing this positive 
chemotaxis in the leucocytes was shown by introducing beneath 
the skin of rabbits tubes filed with such substances as butyrate 
and valerianate of ammonia, trimethylamin, ammonia, glycocoll, 
leucin, tyrosin, urea, etc. These were, for the most part, 
wholly without effect upon the leucocytes, only glycocoll and 
leucin exciting in some cases a moderate chemotaxis, not at all 
to be compared, however, with that of the bacterio-proteins. 



416 THE BACTERIOLOGICAL WORLD. 

It would thus seem to be highly probable, if not absolutely 
proved, that the power of exciting positive chemotaxis, which 
at least many species of bacteria display, is due to the proteid 
ingredients of their bodies when these are set free, as they may 
be naturally when the germs disintegrate in the tissues, or arti- 
ficially by chemical extraction. 

With that keenness and fertility of thought which charac- 
terizes Buchner's work, he now gave wider range to his investi- 
gation. He recognized the fact that, though of late the 
phagocytic action of the leucocytes has been especially empha- 
sized in relation to bacteria, this is by no means their chiefest 
or most constant role. Dispose of bacteria, the leucocytes 
undoubtedly do ; whether after themselves killing them, or 
after they are destroyed b}^ other agencies, has not yet been 
fully determined. But by far the most constant phagocytic 
role of the leucocytes is in carrying on the process of resorption 
and disposal of useless particles and dead material in the living 
body. About such material they gather much as they do in 
the vicinity of bacteria, though not in such marked degree or 
under such dramatic conditions. 

Now, what attracts the leucocytes into the vicinity of a 
particle of dead and useless muscle, or cartilage, or connective 
tissue which they are to absorb and remove ? Certainly not bac- 
terial poison, certainly not bacterial proteids ; for with what 
may be called the normal phagocytic functions of the leucocytes 
bacteria having nothing to do. Having shown that a proteid 
substance derived from the bacterial cells was capable through 
chemotaxis of attracting leucocytes, Buchner now studied in a 
similar way the effects of closely allied substances — namely, the 
so-called vegetable caseins, gluten casein from wheat and legu- 
min from peas, both separated by precipitation from alkaline 
solutions. Both of- these substances were capable of exciting 
the most marked chemotaxis in the leucocytes of rabbits. 
Moreover, as it has been shown that vegetable casein exists as 
such in the grain of cereals and of the leguminosae, he intro- 
duced beneath the skin of rabbits or guinea-pigs, under strict 
antiseptic precautions, masses of wheat and pea meal, and 
found that within two days these masses were surrounded and 
penetrated by enormous masses of leucocytes. Cultures from 



THE BACTERIOLOGICAL WORLD. 417 

these masses proved the entire absence of bacteria. Starch 
introduced subcutaneously under the same conditions induced 
no gathering of leucocytes. 

That this gathering of leucocytes was due to chemotaxis 
and not simply mechanical, owing to the tactile sensibility of 
the leucocytes, was shown by introducing subcutaneously in a 
rabbit in one place an emulsion of infusorial earth with 0.7-per- 
cent, salt solution, and in another place an emulsion of the 
earth with gluten casein. In the first, after three days, but 
few leucocytes had gathered about the foreign material, while 
the second was surrounded and partially penetrated by an enor- 
mous number of leucocytes. 

But still another step remained to be taken. As the gath- 
ering of leucocytes about dead organic fragments in the tissues 
which are to be removed, as so often happens, cannot be ordi- 
narily due to bacteria or bacterio -protein, so, also, interesting 
as the observation may be, can vegetable proteins have no part 
in the matter. So alkali albumenates were prepared and puri- 
fied, in a manner similar to that employed with the bacterial 
and other vegetable proteins from muscle, liver, lungs and 
kidneys of rabbits. These tested in the same way were all 
found to strongly attract leucocytes when introduced beneath 
the skin in tubes. Of the alkali albumenates prepared from 
blood, fibrin, yolk and white of egg, only the blood and yolk 
of egg showed moderate power of exciting positive chemotaxis, 

These experiments show that it is only certain of the 
decomposition products of animal tissue which possess chem- 
otactic powers, and that these, as a rule, are the earlier and not 
the ultimate products of the decomposition. 

Finally, as it has been shown that a general leucocytosis 
is apt to be associated with febrile inflammatory processes. 
Buchner and Roemer studied the effects of iutravenous injec- 
tions in rabbits of these various chemotactic proteids. They 
found that within eight hours of their introduction into the 
blood there was a marked leucocytosis lasting for several hours, 
and that this might be heightened by repeated injections. Thus 
they found, by a daily injection of 2 cc. of an eight-per-cent. 
solution of the bacterio-proteins of bacillus pyocyaneus, the 
relation of white to red blood cells, which at first was 1 to 318, 



418 THE BACTERIOLOGICAL WOKLD. 

was on the second day 1 to 126 ; on the third day, 1 to 102 ; on 
the fourth morning, 1 to 73 : and on the same evening, 1 to 38. 
From this time on no increase was noted. The absolute num- 
ber of the red blood cells remained unchanged, while there was 
an absolute sevenfold increase in the number of leucocytes. 
Gluten casein, as well as alkali albumenate from muscle, 
injected into the blood, showed similar but less pronounced 
effects. 

Considering now the bearing of all these experiments on 
suppuration and on physiological resorption of the dead organic 
materials in the tissues, it would appear that in simple resorp- 
tion, as in bacterial suppuration, the leucocytes are drawn to 
the seat of operation by a proteid material. This in resorption 
seems to be furnished by the dead and disintegrating tissues 
themselves, and when the leucocytes have gathered up a certain 
amount of refuse in their bodies they may carry it away. In 
bacterial suppuration, on the other hand, the attracting 
material may be furnished by the protein of the disintegrating 
bodies of the bacteria themselves, but poisonous ptomaines 
furnished by the live bacteria may cause the destruction and 
degeneration of the attracted leucocytes, which thus collect 
as pus. 

Whether the ptomaines themselves may not indirectly 
furnish chemotactic material by causing the destruction of the 
tissue elements and the setting free of their albuminous con- 
stituents, is a matter requiring further study. 

It is also not improbable that the limited suppuration 
induced by bacteria-free chemical substances — such as turpen- 
tine, calomel, etc., — may be due. to the chemotactic tissue- 
proteids set free by the action of the chemicals on these tissues. 

It seems probable that not only are the leucocytes drawn 
toward the chemotactic proteidsthus produced in or introduced 
into the body, but that the fixed connective-tissue cells are 
stimulated to proliferation. In fact, Buchner found that by 
the introduction of a sterilized emulsion containing 3.5 milli- 
grams of pyocyaneus protein into the forearm of one of his 
associates, a severe inflammation was induced with all the 
symptoms of an acute typical erysipelas, with lymphangeitis, 
such as must have involved the fixed connective-tissue cells. On 



THE BACTERIOLOGICAL WORLD. 419 

the fourth day the m inflammatory process underwent resolution. 
Gluten casein induced similar but less acute effects. 

These most clever and striking researches of Buchner would 
seem to throw much light on the whole subject of the theory of 
suppuration, and to promise large accessions to our knowledge 
of inflammation when the many lines of thought and study 
which they suggest shall have been followed out. 

It is now evident that an aseptic suppuration is possible 
under a variety of conditions. 

It still remains true, however, for the purposes of surgical 
practice, that the suppurative processes as we see them in the 
clinic and at the bedside are due to the presence of bacteria of 
one form or another. It is true, also, that the suppurations 
which we can induce experimentally with sterilized — that is, 
dead — bacterial cultures, or with certain dead proteid substances 
— aseptic suppurations — are limited in their duration, extent 
and destructive power, as compared with those occurring under 
the influence of living germs. This is because in the latter case 
the growing and new forming germs may keep up the inflam- 
mation once alight to an almost indefinite extent. 

We purpose, in the paper which is to follow, to detail some 
results of a series of experiments on the action of dead tubercle 
bacilli on the living tissue not less striking than are those which 
show the power of other sterilized bacteria to induce suppura- 
tion. — Neto Yorfc Medical Journal. 



THE CHEMICAL FACTORS IN THE CAUSATION OF DISEASE. 



LECTURE DELIVERED AT THE POST-GRADUTE MEDICAL SCHOOL OF CHICAGO 
BY VICTOR C VAUGHAN, M. D., PH. D., OF ANN ARBOR, MICH. 

Professor of Hygiene and Physiological Chemistry and Director of the 
Hygienic Laboratory in Michigan University. 



LECTURE I. GENERAL CONSIDERATIONS OF THE RELATION OF 
BACTERIAL POISONS TO THE INFECTIOUS DISEASES. 

The majority of diseases may be grouped, from an etiologi- 
cal standpoint, into the following classes : First, traumatic ; 
second, infectious ; third, autogenous, and, fourth, neurotic. 



420 THE BACTERIOLOGICAL WORLD. 

It must be understood, however, that in many diseases the 
cause is not single, but multiple, and for this reason sharp lines 
of classification cannot be drawn. For instance, the greatest 
danger in those traumatic affections in which the traumatism 
itself does not cause death, lies in infection. The wound has 
simply provided a suitable point of entrance for the infecting 
agent. Indeed, the break in the continuity of tissue may be so 
slight that it is of import and danger only on account of the 
coincident infection. This is true in many cases of tetanus. 
Furthermore, an infectious disease, whether it originates in a 
traumatism or not, is markedly influenced by what we are 
pleased to call the idiosyncrasy of the patient, and by which 
we mean the peculiarities of tissue metabolism taking place in 
the individual at the time. A dozen men may be exposed alike 
to the same infection, and the infecting agent may find a suit- 
able soil for its growth and development in two of these, while 
in the other ten this same agent meets with such adverse influ- 
ences that it dies without producing any appreciable effect ; or 
all may be infected, but with differences in degree, as is evi- 
denced by variations in symptoms, in the length of time that 
the infecting agent continues to grow and develop in the body, 
and in the ultimate result. Every physician who has had 
experience in the treatment of typhoid fever, diphtheria, 
scarlet fever — or, in short, of any of the infectious diseases— will 
appreciate the importance of the personal equation in his 
patients. That some neurotic affections originate in trauma- 
tism, we know. That others of this class are largely due to 
malnutrition accompanied by improper metabolism or insuffi- 
cient elimination ; or, in other words, are to some extent auto- 
genous, all believe. Understanding, then, that the above 
classification does not attempt a sharp and marked differentia- 
tion of the causes of disease, I wish to devote the first two of 
the three lectures, which you have kindly asked me to give, to 
a consideration of the chemical factors in the causation of the 
infectious, and of the traumatic, autogenous and neurotic dis- 
eases in so far as these are influenced by infection. 

The science of bacteriology is now entering upon a new 
and promising era in its development. Heretofore, this science 



THE BACTEKIOLOGICAL WOKLD. 421 

has been largely founded upon morphological studies. Bac- 
teriologists have given their time and attention to the discovery 
of bacterial forms in the diseased organism, and to the observa- 
tion of characteristics in structure and growth of different 
species of bacterial life. The question, Do certain germs have 
a causal relation to certain diseases ? having been settled in the 
affirmative, the next question which naturally arises is, In 
what way is this causal effect accomplished ? How do germs 
prove harmful ? To this question, a number of answers have 
been proposed. But as I have elsewhere* discussed these 
theories, I will not repeat them here. Suffice it to say that it 
is now generally admitted that the deleterious effects wrought 
by germs are due to chemical poisons elaborated by them dur- 
ing their growth. Granting this, it will be at once seen that 
the morphological study of germs, important as it is, becomes 
wholly inadequate in ascertaining their true relationship to the 
diseases with which they are associated. We must now study 
the physiology and the chemistry of germs, and until this is 
done we must remain ignorant of the true cause of disease, and, 
so long as we remain ignorant of the cause, it cannot be expected 
that we shall discover scientific and successful methods of 
treatment. Suppose that our knowledge of the } 7 east plant was 
limited to its form and method of growth, of how little practi- 
cal importance this knowledge would be. That the yeast 
plant requires a saccharine soil before it can grow, and that 
given such a soil it produces carbonic acid gas and alcohol, are 
the most important and practical facts which have been 
ascertained in its study. Likewise, the conditions under which 
pathogenic germs multiply and the products which they 
elaborate in their multiplication must be ascertained before 
their true relationship to disease can be understood. 

In saying that the morphological work upon which the 
science of bacteriology rests almost wholly is inadequate, I 
wish that it may be plainly understood that I am not offering 
any hostile criticism upon the great men who have done this 
work and who have formulated conclusions therefrom. The 
development of bacteriology has been in accordance with the 
natural laws governing the growth of all the biological sciences. 

* Ptomaines and Leucomaines. 



422 THE BACTERIOLOGICAL WORLD. 

The study of form naturally and necessarily precedes the study 
of function. The ornithologist finds a new species of bird. He 
first studies its shape and size, the color of its plumage, the 
form of its beak, the number and arrangement of the feathers 
of the tail and. wing, the color of the eyes, etc. All this he 
can do with a single specimen, recognizing the fact, however, 
that variations, more or less marked, are likely to be found in 
other individuals. More time and wider opportunities of 
observation will be needed before he can tell where and. when 
this bird is accustomed to build its nest, upon what insects, 
grains and. berries it feeds, with what other species of birds it 
lives in peace and with what it is at war. A much greater 
range of observation and study is necessary before the naturalist 
can tell how his newly discovered species would thrive if carried 
to a new climate, where it would be compelled to live upon 
unaccustomed food, to build its nest of strange material and 
to encounter new foes. I repeat that it is no discredit to the 
science nor to the men who have developed it, to say that the 
study of bacteriology has hitherto been almost wholly morpho- 
logical ; without the morphologist, the physiologist and the 
physiological chemist would not exist. The science having 
had for its support only morphological studies, the deductions 
and formulated statements arrived at by its students have been 
reached in accordance with the knowledge obtained from this 
source. But now, it being admitted that the causal relation 
between a given germ and a certain disease is dependent upon 
the chemical products of the growth of the germ, the funda- 
mental lines of work must be altered in order to correspond 
with this new knowledge. Let us inquire into the changes, 
which the introduction of this new factor must make in our 
fundamental conceptions of the causal relation between germs 
and disease. 

The four rules of Koch have been generally conceded to 
be sufficient to show that a given germ is the sole and sufficient 
cause of the disease with which that germ is associated. 
Briefly these rules are as follows : 

First. The germ must be found in all cases of this disease. 

Second. The germ must be separated from other organ- 
isms and from all other matter found with it in the diseased 
animal. 



THE BACTERIOLOGICAL WORLD. 423 

Third. The special bacterium thus freed from all foreign 
matter must, when properly introduced, produce the disease in 
health} 7 animals. 

Fourth. The micro-organism must be found properly dis- 
tributed in the animal in which the disease has been induced. 

Let us give our special attention to the first of these rules 
for a few moments. What is meant by the statement that the 
special germ must be found in every case of the disease ? How 
will A, pursuing his studies on the bacteriology of a given dis- 
ease in America, decide whether or not a bacillus, which he 
finds, is identical with one which has been reported as invari- 
ably present in the same disease, by B, who has investigated an 
epidemic in Germany ? What means are relied upon to prove 
the identity of these two organisms ? The means which have 
been relied upon wholly are the form, size, reaction with stain- 
ing reagents, manner of growth on the various nutrient media, 
and, in exceptional instances, correspondence in their effects 
upon the lower animals. In other words, with the exception of 
those instances in which the effects upon animals are tried, the 
characteristic, by which the germ causes the disease, is left 
wholly out of consideration. 

Indeed, some of the most eminent bacteriologists have 
taught that in the identification of germs the reactions with 
staining agents and the appearances of the growths on the 
various nutritive media are of more importance than the obser- 
vation of the effects upon animals. Thus, Fluegge says : 

" Inoculation experiments with both typhoid dejections and pure 
cultures of the Eberth bacillus have universally been without success. 
'The few experiments in which a typhoid disease has followed inocula- 
tion or feeding have been made with impure material containing other 
active bacteria. It is known that a group of widely distributed organ- 
isms, which, however, are wholly different from the typhoid bacillus, 
have the power, when injected subcutaneously or intravenously, of 
producing in animals death with marked swelling and ulceration of 
Peyer's patches. To these organisms undoubtedly are due the appar- 
ently positive results which some authors have supposed to be due to 
inoculation with the typhoid bacillus." 

In other words, this eminent author teaches that, although 
other germs may cause the essential symptoms and lesions of 
typhoid fever in the lower animals, they are not related to the 



424 THE BACTERIOLOGICAL WORLD. 

germ found in the spleen of man after death from typhoid 
fever, because they do not react in the same manner with the 
aniline stains. 

Bacteria cannot be classified so far as their causal relation- 
ship to diseases is concerned ( and this is the most important 
knowledge to be gained from them ) until we know the nature 
of the chemical poisons which they elaborate, for it is by virtue 
of these poisons that they have any causal relationship to 
disease. 

It is possible that two germs may be unlike in form, and 
yet they may produce poisons which are identical or those which 
are very similar in their effects upon man. One germ may be 
stained by Gram's method and another fail to be acted upon 
when so treated, but this does not prove that their chemical 
products are totally unlike. 

We will suppose that in an epidemic of diphtheria A 
examines the membrane from a hundred, or we might as well 
suppose a thousand, children and finds a characteristic, well- 
marked, easily recognized bacillus in all. He isolates this 
organism and obtains it in pure culture. He inoculates animals 
and these manifest all the signs together with the appearance of 
the characteristic membrane of diphtheria, and in these animals 
he finds his bacillus growing as in the throats of the children. 
All of the rules of Koch have been complied with. Has A 
demonstrated that his bacillus is the sole cause of diphtheria ? 
ISTo. He has shown that his bacillus is a cause of diphtheria, 
but he has not proven that there may not be other germs, wholly- 
different from his in form and size, which may also cause diph-. 
theria. The most which can be proven by Koch's rules is that 
a given germ is a cause of the disease. They do not <show 3 as 
most bacteriologists would have us believe, that this germ is the 
sole cause of the disease. 

To illustrate, we will suppose that a botanist in visiting 
Arabia should find a tree producing a berry, the coffee berry, 
which when properly prepared and taken into the system pro- 
duces certain effects. These effects are due to the alkaloid 
caffein, and our supposed discoverer finds that they invariably 
follow the drinking of a decoction of these berries. Would he 
be justified in concluding that the coffee tree is the only plant 



THE BACTERIOLOGICAL WORLD. 425 

in the world capable of producing these supposed characteristic 
effects ? Should he reach such a conclusion, the fact that it is 
not warranted would be shown by a study of the tea plant in 
China and the guarana of South America. 

What is meant in the first of Koch's rules by the statement 
that the germ must be present in all cases of the disease? In 
answering this question, I will give two specific illustrations 
showing the manner in which eminent bacteriologists have 
applied this rule. This is well shown in the bacteriological 
literature of typhoid fever and diphtheria. 

Eberth found the bacillus, which is now generally consid- 
ered by bacteriologists as the sole cause of typhoid fever, in 
eighteen out of forty cases. In the examination of twenty- 
eight bodies dead from typhoid fever .Gaffky found the Eberth 
germ in twenty-six ; Fraenkel and Simonds reported in twenty- 
five out of twenty-nine cases ; Seitz in twenty-two out of 
twenty-four ; Rietsch in thirty -five out of thirty-six ; and 
Kowalskski found it present in each of twenty-nine cases. From 
these and similar reports, bacteriologists have concluded that 
this germ is not only a cause, but the sole cause of typhoid fever. 
Are they justified scientifically in this conclusion? Have they 
any right to say that there is no other germ in existence which 
may cause typhoid fever? The speaker has shown that there 
are at least two other germs which induce the same symptoms 
in a more intensified form in the lower animals, and has iso- 
lated and studied the chemical poisons of these germs, and 
recenty Babes has shown that in the examination of twelve per- 
sons who died at Bucharest of typhoid the Eberth germ was 
not present, and that an organism giving quite different reac- 
tions was present. 

Loeffler and other German investigators have shown the 
presence of a bacillus in the majority, not in all, of the cases of 
diphtheria which they have studied, and they again conclude 
that this bacillus is not only a cause but the sole cause of the 
disease. In a large number of cases in New York, Prudden 
failed to find the Loefner bacillus, but did find a wholly different 
organism. ~No one acquainted with Prudden's work will ques- 
tion the accuracy and care with which it was done. It is true 
that the chemical poisons of the Loefner bacillus have been 



426 THE BACTERIOLOGICAL WORLD. 

studied, and no one can doubt that it is a cause of diphtheria, 
but it may be found that the streptococcus of Prudden is capable 
of elaborating a similar poison. 

The moment that it is granted that the real poison of the 
disease is chemical in character, it becomes evident that no one 
is justified in saying that one germ is the sole source of such a 
poison. Such a statement would be as much unwarranted as 
one that the coffee tree is the sole source of caffein, or that the 
strychnos ignatii is the only species of the natural order log- 
aniacece which produces a convulsive poison. In other words, 
the specific cause of a given disease is not to be determined 
wholly by the morphology of the germ found associated with 
that disease in a given epidemic, but by the character of the 
chemical poison which is the true maleries morbi. 

I think that we are justified in concluding that, in those 
diseases in which the four rules of Koch have been complied 
with, the germ is a cause of the disease, but our range of obser- 
vation must be much wider than it now is, before we can say 
that the given germ is the only cause of the disease. It may be 
found, and I think this highly probable, that those few infec- 
tious diseases, such as anthrax and tuberculosis, which have 
such well-marked typical clinical histories, are due to equally 
well-marked and morphologically distinct micro-organisms 
which can be recognized by microscopical study alone ; but I do 
not expect that this will prove to be true in diseases showing 
such wide variations in symptoms as is doue by typhoid fever 
and cholera infantum. In fact I am convinced that these last- 
mentioned diseases may and do result from any one of a number 
of bacteria which are in form, size, methods of growth and 
reactions towards staining reagents quite distinct one from the 
other, but which produce poisons similar in their effects. 

That you may be convinced of the truth of the foregoing 
statements I give a few specific instances which will confirm 
them, and demonstrate that, in their causal relation to disease, 
germs cannot be classified from a study of the form, size, 
methods of growth and reactions with staining reagents. 

Typhoid fever. — Eberth lays special stress upon the fact 
that his germ does not take the ordinary stains readily, and 
says that it is not colored by Bismark-brown and hasmotoxylin, 



THE BACTERIOLOGICAL WORLD. 427 

and only slightly by the other ordinary stains ; but what is the 
verdict of others on this point? In the paper in which he calls 
attention to the fact that he has photographed the short bacillus 
before Eberth had published his discovery, Koch says : "Eberth 
states that these short bacilli have but slight tendency to take 
stains. The photograph here given shows, to the contrary, 
that in taking stains these bacilli are but slightly inferior to 
other bacteria/'' G-affky states that the best coloring agent is 
methyl-blue, that the bacilli are stained very well with methyl- 
violet, gentian-violet, Bismark-brown and fuchsin, and less well 
by haemotoxylin. Coates had no difficulty in staining with 
Bismark-brown. Meyer had the same difficulty as Eberth in 
staining ; but Eriedlander found that the bacilli in sections 
from the spleen stained intensely. In view of this variety of 
statement, can anyone claim that these men were working with 
a germ which can be distinguished from others by its tinctorial 
properties ? 

Tetaxus. — The tetanus germ of Kitasato and that of 
Tizzoni and Cattani are known to be quite distinct. Cultures 
of the former in bouillon are virulent, while those of the latter 
in the same medium are inert. Not only are these two organ- 
isms morphologically and biologically different, but their poisons 
are chemically unlike. Brieger and Fraenkel precipitated the 
poisonous albumen of the germ of Kitasato with alcohol, but 
this reagent renders the poison of the Italian germ inert. 
Notwithstanding this difference, however, both micro-organisms 
and their chemical products produce tetanic convulsions and 
death in the lower animals. TTe must, therefore, admit that 
there are at least two distinct germs, each of which is capable 
of causing tetanus, and how many other bacteria with like 
properties there may be no one can tell. 

The summer diarrhoeas oe ixfaxct. — In this class of 
diseases all attempts to find a morphologically specific germ have 
failed. The labors of Booker in this country, and of Escherich 
in Germany, have shown that no one species or variety is con- 
stantly present. No less than thirty distinct germs have been 
obtained from the bowels and faeces of children suffering from 
the summer diarrhoeas. A germ which is frequently present 
one season will not be found at all the next. Are we to conclude 



428 THE BACTERIOLOGICAL WORLD. 

from this failure to comply with the first of Koch's rules that 
the summer diarrhoeas of infancy are not due to micro-organ- 
isms ? Certainly not ; especially in view of the fact that 
Baginsky and Stadthagen have shown that the bacillus discov- 
ered by Baginsky produces a chemical poison which induces 
diarrhoea in the lower animals, and I have shown that at least 
three of Booker's bacteria produce poisons which have similar 
effects. To the contrary, we are justified in concluding that 
the diarrhoea may be due to any one or more of a number of 
germs which differ from one another sufficiently morphologic- 
ally to be classified as distinct species. The similarity among 
these bacteria will not be discovered by a study of their size, 
form and reactions with staining reagents, but by a study of their 
chemical poisons, the agents by virtue of which they cause the 
disease. At the meeting of the American Medical Association 
at Newport, in 1889, I proposed, in a paper on the etiology of 
typhoid fever, that in those cases in which the lower animals are 
not available, on account of their insusceptibility, before a given 
germ can be considered a cause of a disease, it must be shown 
that that germ can produce chemical poisons which will induce 
in the lower animals, in an acute form, the characteristic 
symptoms of the disease. That this rule would be a wise one 
I have become, even more strongly than I was at that time, 
convinced. 

In the little volume on " Ptomaines and Leucomaines," 
written by Novy and myself, and published in 1888, I gave the 
following definition of an infectious disease : i{ An infectious 
disease arises when a specific, pathogenic micro-organism, hav- 
ing gained admittance to the body and having found the con- 
ditions favorable, grows and multiplies, and in so doing 
elaborates a chemical poison which induces its characteristic 
effects." 

It should be remembered that this definition was written 
at a time when the importance of the chemical products of bac- 
terial growth was not generally recognized. Indeed, at that 
time bacteriologists quite universally held to the mechanical 
interference theory, and more than a year later so good an 
authority as Welch, of Baltimore, stated that a knowledge of 



THE BACTERIOLOGICAL WORLD. 429 

the chemical products of bacteria was interesting but not essen- 
tial to a belief in the causative agency of a micro-organism, and 
he went still further, inasmuch as he claimed that the success- 
ful inoculation of animals is not necessary in order to prove the 
causal relationship of a certain germ to a given disease. Now, 
it is generally conceded that a successful inoculation is a proof 
of the formation of deleterious chemical products, and in case 
successful inoculation cannot be obtained on account of the 
insusceptibility of the lower animals, the demonstration of the 
elaboration of chemical poisons is deemed not only interesting 
but important. For instance, the strongest proof that the 
cholera bacillus of Koch has a causal relation to Asiatic cholera 
has been furnished by the study of its chemical products, and 
the similar studies of many of the other infectious diseases 
have yielded results which are of both interest and importance, 
as I shall endeavor to show in the next lecture. 

The only modification which I desire to make in the above 
definition of an infectious disease consists of an elaboration of 
what is meant by a " specific " micro-organism. The specific 
nature of a germ is to be determined not by its size, form? 
growth on nutritive media and reactions with staining reagents, 
all of which properties are more or less variable with the con- 
ditions under which the germ has been grown, but upon its 
capability of producing within the body deleterious substances. 

The study of the chemical factors in the causation of the 
infectious diseases opens up for us a field in which much work 
must be done. Let us attempt a statement of the nature 
of some of the researches that must be carried out along this 
line. 

In the first place we must ascertain what germs are toxico- 
genic. This would necessitate a chemical study of all kinds of 
bacteria, both the pathogenic and the non-pathogenic. Every 
fact learned in this investigation will not have its practical 
application in medicine, but will have its scientific value, and 
many will most probably be of more or less direct service to 
man. There is a vast field here for careful scientific work, and 
I should be glad to see these researches carried out largely in our 
own country. 



430 THE BACTERIOLOGICAL WORLD. 

Secondly, it must be determined under what conditions 
these germs are toxicogenic. It is not at all probable that all 
those bacteria which are capable of producing poisons when 
grown on dead material outside of the body are also capable of 
multiplication and the production of the same substances when 
under the influence of the various secretions of the body. Some 
bacteria are destroyed by a normal gastric juice within a short 
time, while others are not. The conditions of life and growth 
are different when the infecting agent is introduced into the 
blood from what they are when infection occurs by the way of 
the alimentary canal. This is well recognized in the two forms 
of anthrax, one of which arises from inoculation through a 
wound and the other by way of the intestines. A preventive treat- 
ment which is efficient in the one is of no service in the other. 
Then, again, we are to study those conditions of the blood and 
other fluids of the body, which are especially unfavorable to the 
successful implantation or the continued existence of an infec- 
tious disease. 

Thirdly, the chemical properties and the physiological 
action of these poisons will demand careful attention. Some 
are especially depressing in their action upon the heart, others 
seem to manifest their energy upon the central nervous sys^ 
tern, while others still act like true gastro-intestinal irritants. 
In the study of the toxicological effects of these bacterial 
poisons every method of investigation known in the most 
advanced physiological work must be employed. The action of 
these agents on the heart, the brain, the spinal cord, etc., must 
be thoroughly studied. 

After all this has been done we shall certainly advance in 
our knowledge of the treatment of the infectious diseases. The 
morphological study of bacteriology has been of inestimable 
value to us in the prevention of the infectious diseases. It has 
rendered the investigation of disinfectants both possible and 
imperative, and by the knowledge thus gained yellow fever has 
repeatedly, and Asiatic cholera in one instance at least, been 
kept from our shores. Epidemics of small-pox, diphtheria and 
scarlet fever have been arrested. In this way scientific medi- 
cine has been advanced, and such an advance is never made 



THE BACTERIOLOGICAL WORLD. 431 

without great benefit to mankind in general. Moreover, anti- 
septic surgery, which had already been inaugurated by the 
genius of Lister, has received the scientific support which was 
necessary to its constant, universal and correct employment. 
All of these benefits, and possibly more, which do not occur to 
me at this time, have been accomplished, and to Pasteur, Koch 
and the host of others who have rendered this possible, we 
should be and are grateful. But what can those of us who are 
engaged in the treatment of the infectious diseases say of the 
present condition of the practice of medicine ? As one of this 
number I am forced to admit that my successes are few and my 
failures many. Are we satisfied with the treatment which we 
now employ for tuberculosis, diphtheria, scarlet fever, typhoid 
fever and the other internal infectious diseases ? Do we treat 
these diseases with much more success than our fathers did ? 
They prescribed for their consumptive patient liberal food, cod- 
liver oil, and all the open air exercise possible. They never 
carried a Begeron hydrogen-sulphide bag, used a Weigert hot- 
air oven, or injected Koch's lymph ; but can we say that our 
successes are markedly more numerous than theirs ? Has the 
knowledge of germs aided us in the treatment of these diseases ? 
Do we to-day possess one germicidal agent which will destroy the 
bacilli of typhoid fever in the spleen of our patient and at the 
same time do no injury to the patient himself ? 

For myself I am perfectly willing to acknowledge that I am 
not satisfied with my success in the treatment of the infectious 
diseases, and it seems to me that careful scientific study along 
the lines which we have been following to-night holds out suffi- 
cient encouragement to lead us to further study. Could we 
treat morphine poisoning before we knew anything of the chem- 
ical properties or the physiological and toxicological effects of 
this alkaloid ? Can we counteract or antidote the typhoid 
poison, before we know anything of its nature or its manner of 
action ? I do not mean to imply that it is the duty of each of 
us to become a chemist or to undertake physiological research. 
We need as much as anything else a closer study of these sub- 
jects at the bedside. The scientific methods of the laboratory 
should be carried into the wards of the hospital. The phy- 
siologist keeps the dog upon which he is making some import- 
ant study under his own trained eye and trusts no one else with 



432 THE BACTERIOLOGICAL WORLD. 

the observations. The patient with typhoid fever in the hospi- 
tal ward is seen by his medical attendant once or twice during 
the day. The remainder of the time all observations must be 
made and records taken by a nurse, who is not only devoid of 
scientific training, but has. so much work to do that but little 
time can be given to the individual patient. There is work 
enough in the study of the poisons of disease to employ the 
chemist, physiologist and clinician, and the greatest good can 
be secured only by the combined labors of all. 

You may think it strange that, enthusiastic as I am about 
the importance of the study of the bacterial poisons, I have said 
nothing about the therapeutical uses to which some of them 
may be put, as evidenced by the introduction by Koch of his 
so-called lymph or tuberculin as a cure for tuberculosis. 
There may be valuable curative agents discovered among the 
products of bacterial activity, but I have never believed that a 
disease will be cured by the administration of the very poison 
which is causing the disease. I say I have never believed this, 
but I am open to conviction on this point, and take much 
interest in a study of the results now being reported from all 
parts of the world, as well as in my own observation of the 
effects of the proposed cure. So far, however, I have seen no 
sufficient reason for any change in my belief on this point. 

( To he continued.) 



THE BIOLOGY OF THE LANCEOLATE DIPLOCOCCUS. 
[TRANSLATED BY H. W. LOEB, A. M., M. D., ST. LOUIS, MO.] 

by pio foa (Turin). 

The diplococcus lanceolatus appears somewhat different if 
taken from an inflamed lung or from meningitic pus. The 
extraordinarily different frequency of occurrence of pneumonia 
and cerebro-spinal meningitis encourages the thought that the 
two diseases may differ from each other in the action of the 
micro-organisms. However, the writer was unable to obtain 
any change in the virulence by transplantation of the cultures. 



THE BACTERIOLOGICAL WORLD. 433 

Some little is observed, depending upon the course and age of 
the cultures. The strength of the virulence, therefore, depends 
upon the shorter or greater persistance of the cocci upon the 
inoculated part. Besides this, by the conjoined inoculation 
with the staphylococcus or with the proteus vulgaris the quali- 
ties of the pneumo-coccus may be given to the meningo-coccus. 
The writer discusses further the differences in the ana- 
tomical results which occur in the organs of the body when 
they are inoculated with the meningo-coccus or pneumo-coccus. 



THE HISTOLOGICAL CHANGES OF THE ORGANS IN DIPH- 
THERIA. 

[TRANSLATED BY H. W. LOEB, A. M., M. D.] 

by babes ( Bucharest). 

The writer describes the tissue changes in diphtheria, 
which entirely correspond to the description given by Oertel. 
So far as the relation of these changes to the Loeffler bacillus is 
concerned, the difficulty of interpretation proceeds from the 
fact that streptococci accompanying the bacillus of Loeffler, as 
well as other bacilli which are similar, though not identical, to 
those of Loeffler. 

He mentions the remarkably great caryokinesis which is 
found in the diphtheritic inflamed membranes. On account 
of the chemical products of the diphtheria bacillus one can only 
very imperfectly produce the peculiar anatomical changes of 
diptheria, whereas the picture which is obtained by the inocu- 
lation of the bacilli is on the whole very identical to human 
diphtheria. 

( To be continued.) 



434 TB*E BACTERIOLOGICAL WORLD. 



SCIENTIFIC GLEANINGS. 



BACTERIAL POISONS. 



BY VICTOR C . VAUGHAN . 

Medical News ; Popular Science News. 

The author sums up the present state of our knowledge on 
the above subjects as follows : 

First. Man is attacked by the infectious diseases either 
through the alimentary canal or through the blood or lymph. 

Second. The gastric juice is a physiological guard against 
infection by the way of the intestines. 

Third. Additional guards against infection by the intes- 
tines are probably to be found in the absorbing cells of the 
stomach and intestines. 

Fourth. Susceptibility to the intestinal infectious diseases 
is increased when, for any reason, these physiological guards 
are defective. 

Fifth. All toxicogenic germs are dangerous when intro- 
duced into the intestines, and their capability of doing injury 
lies in their production of chemical poisons. 

Sixth. Many of these poisons are proteid in character. 

Seventh. These poisonous proteids most probably act by 
catalysis. 

Eighth. In the splitting up of complex molecules into sim- 
pler ones, heat is liberated and fever manifests itself. 

Ninth. The physiological guard against infection through 
the blood or lymph lies in the germicidal action of the proteids 
of these fluids. 

Tenth. Susceptibility to infection through the blood or 
lymph is increased by impoverishment of these fluids. 

Eleventh. We can continue to treat consumption and 
other systemic diseases by the employment of liberal diet, 
exercise in the open air, and constitutional remedies without 
being unscientific in our practice. 



THE BACTERIOLOGICAL WORLD. 435 

Twelfth. Filth, without being the bearer of a specific germ, 
is a cause of disease. 

Thirteenth. Wherever man pollutes the soil about him, 
the air which he breathes, and the water which he drinks with 
his own excretions, .there enteric fever will be found. 

Fourteenth. In their causal relation to disease, germs can- 
not be classified without a knowledge of the chemical changes 
which they induce. 

Fifteenth. While certain bacterial poisons can result only 
from the growth of certain germs, other poisons similar to one 
another in their action, though probably not identical, may 
result from any one of a number of organisms. In the former 
case we have such diseases as anthrax and small-pox, with their 
practically constant symptoms and well-marked course ; in the 
latter case we have such diseases as the summer diarrhoea of 
infancy and enteric fever, with their varying symptoms. 
— Buffalo Medical and Surgical Journal. 



BACILLUS OF CHOLERA IN SOIL. 

Prof. V. de G-iaxa draws the following conclusions from 
his experiments with the cholera bacillus on soil. When the 
cholera bacillus enters a soil rich in common bacteria, even 
though it finds conditions favorable to its existence and repro- 
duction, it rapidly succumbs in the struggle which takes place 
between itself and the other bacteria. Tbe latter increase in 
number, and this increase is rendered possible as far as the 
deeper layers of the soil are concerned by the addition of nutri- 
ent material which agrees with them, and also modifies the 
condition of the ground. 

Should the cholera bacillus enter in relatively large num- 
bers a soil which is inhabited by a few ordinary bacteria, not 
only its preservation, but also its reproduction become possible, 
until an increase of the common bacteria takes place from the 
penetration of the soil by nutritive matters which enter along 
with the pathogenic bacteria. 



436 THE BACTERIOLOGICAL WORLD. 

t 

MICRO-ORGANISMS OF THE HEALTHY STOMACH AND 

THEIR ACTION. 

M. J. E. Abelous confirms the results of Pasteur and 
Duclaux relative to the action of microbes in the stomach, and 
the part they play in digestion. The microbes were taken 
from the author's stomach by washing it out after fasting. The 
usual precautions were taken to prevent external contamina- 
tion, and the mouth and pharynx were previously washed out 
with sublimate. The microbes were cultivated on gelatin, 
peptonized and glycerined gelatin, potato, gelatinized serum, 
neutral and acidulated bouillon. The species 'isolated were 
sar cina ventriculi bacillus pyocyaneus, bacterium lactis aero- 
genes, bacillus subtilis, bacillus mycoides, bacillus amylobacter, 
vibrio rugula, and nine other species, which are distinguished 
by letters, A, B, 0, etc. These include one coccus and eight 
bacilli* The chief facts about these are that their resistance to 
an artificial gastric juice is greatly in excess of the mean dura- 
tion of digestion in the stomach, and that they are potentially 
anaerobic. — Exchange. 



TECHNIQ UE. 



EXAMINATION OF WATER FOR TYPHOID BACILLUS. 

Dr. Finklenburg (Centralbl. f. Bakt. u. Parasitenk., Bd. 
ix, No. 9, 1891) points out that in examining water for typhoid 
bacillus the sediment should always be used as well as the water 

itself. In a certain village, X , there was an outbreak of 

typhoid fever amongst the boys of the school, following a simi- 
lar outbreak in a neighboring group of houses. There was 
found to be a dangerous proximity of the spring from which 
the water was derived to the closets in the schoolyard. Exam- 
ining the water by the ordinary gelatine plate method, the 
author was unable to find any traces of the Eberth-Garrky bacil- 
lus in any of his cultivations ; but on allowing the water to 
stand and making cultivations from the sediment, there 
appeared, among others, characteristic colonies, the bacilli of 



THE BACTERIOLOGICAL WORLD. 437 

which, in staining and microscopic appearances, could not be 
distinguished from those of the typhoid bacillus. 

The author considers that this examination of deposited 
matter should never be omitted, as he thinks that it is probable 
that many of the germs must be held mechanically suspended 
with the solid constituents in the water rather than free in the 
water itself, as these organisms are almost invariably intro- 
duced along with solid sewage or other organic matter. He 
thinks such examination is especially necessary wherever a 
search is made for pathogenic micro-organisms. — Supp. British 
Medical Journal. 



A REVIEW OF THE METHODS OF DEMONSTRATING THE 

FLAGELLA ON MOTILE BACTERIA WITH SPECIAL 

REFERENCE TO THE STAINING PROCESS* 

BY VERANUS A. MOORE, M. D. 

Assistant in the Laboratory of the Bureau of Animal Industry, 
Department of Agriculture, Washington, D. C. 

There is, perhaps, no element in the known structure of 
micro-organisms that is more difficult to demonstrate than the 
flagella of motile bacteria. Ehrenberg was among the first of 
the earlier investigators in this field to infer from the discovery 
of a vortical action in front of a large motile spirillum ( s. volu- 
tans) that its movements were produced and controlled by a pair 
of fine flagella, one at each end of its spiral body. With the 
improved microscopical apparatus, Oohn ( 1 ) succeeded in 
demonstrating the presence of the supposed flagella on this spi- 
rillum, and his discovery was confirmed shortly afterwards by 
Dallinger, Drysdale, and others. This fact suggested the idea 
that all motile bacteria depended for their movement on similar 
motile filaments. 

Dallinger and Drysdale ( 2 ), basing their work upon the 
fact that the minuter monads are provided with one or more fla- 
gella, turned their investigations to bacterium termo to deter- 
mine whether or not there existed a flagellum or flagella on that 



* Read before the Washington Microscopical Society, February 20, 1891. 



438 THE BACTERIOLOGICAL WORLD. 

bacterium. For this work a -^ inch Powell and Lealand 
immersion lens were employed, and special care taken to secure 
the best possible illumination. The specimen examined was 
prepared by adding a bit of culture of the bacterium termo 
grown in Cohn's nutritive fluid to a drop of distilled water on a 
slide and covering it with an extremely thin cover glass. The 
examination was continued incessantly for nearly five hours, 
when a flagellum was distinctly seen at one end of each of two 
termo that were moving slowly across the field. The flagella, 
as seen on the moving germs, were exquisitely delicate and con- 
stantly lashing. Other large motile bacteria were subsequently 
found to possess similar flagella. 

The difficulty in detecting the flagella on motile bacteria in 
a fresh condition is well illustrated by the above briefly- 
described experiment. It is also a fact worthy of note that the 
flagella have been seen by this process only on the larger sapro- 
phytic forms, and that the smaller and especially the pathogenic 
germs could not be considered as possessing flagella, except from 
analogy, until other methods had been devised. 

Dr. R. JSTeuhauss ( 5 ) reports a successful demonstration of 
the flagella on the comma bacillus by means of photography. A 
culture of this bacillus in meat broth was allowed to grow for 
four weeks, when it showed, instead of the ordinary delicate 
comma bacilli, large, thick bacilli and long spirilla. Most of 
these had lost their motility, but a few germs were still capable 
of movement. A preparation was made by adding a small quan- 
tity of this culture to a drop of distilled water on a slide and 
covering it with a thin cover glass. In this condition they were 
photographed, and a negative obtained which showed a delicate 
spiral flagellum attached to a short, much-curved bacillus. 
Other successful negatives were also obtained from the same 
field by focusing. 

It is, however, through the development of staining 
methods that the demonstration of these hair-like appendages 
on the great majority of motile bacteria has been made possible, 
and, in many instances, comparatively simple. The discovery 
of a process by which these minute structures could be stained 
was first made by Koch, who, after a long series of negative 
experiments made with the ordinary dyes employed in staining 



THE BACTERIOLOGICAL WORLD. 439 

bacteria, finally succeeded in staining the flagella on a certain 
number of saprophytic forms by using a solution of the extract 
of logwood. Since then other methods have been devised for 
staining the flagella on pathogenic as well as saprophytic bacte- 
ria, so that now we are able to demonstrate their presence on 
nearly all known motile germs. The development of these 
methods has been attended with much interest for each step 
in advance has been taken only after long and patient study, 
aud each in its turn has been to a greater or less degree the 
foundation for the succeeding one. In reviewing the various 
methods, I shall give them in a form as condensed as possible, 
assuming that the ordinary technique in staining bacteria is 
somewhat familiar. It is but just, however, to note that each 
author to be referred to gives a very full account of his experi- 
ments, and, in nearly every case, the experiments and results 
which led to the formulation of the method finally recom- 
mended. Although of much interest in the history of the 
development of the latest methods, the unsatisfactory processes 
will, as a rule, be omitted at this time. 

Koch's method. — Koch ( 4 ) employed as his staining fluid 
a concentrated aqueous solution of the extract of logwood. 

The bacteria and" their flagella were stained by applying this stain- 
ing fluid in two different ways : First. By adding the solution to a small 
quantity of the germ-containing liquid; second, by making cover-glass 
preparations, by the ordinary method, from the germ-containing sub- 
stance, allowing them to dry in the air, after which the film on the 
cover glass was covered with the staining solution and allowed to 
remain for a considerable length of time. The cover was then rinsed 
in water and the preparation mounted in glycerine. The flagella were 
stained a beautiful brown. In order to retain the flagella in a stained 
condition the preparations were treated, prior to mounting, with a 
weak solution of chromic acid or with Mueller's fluid. The stained 
cover glass could he allowed to dry and mounted in balsam. By this 
process Koch preserved a specimen of the bacillus tremulus which 
showed both the spores and flagella. 

Xeuhauss' method (5). — This method was devised with 
special reference to the staining of the flagella of the comma 
bacillus. It is as follows : . 

The dried' cover-glass preparations of a culture of this germ were 
boiled five minutes in common (Kaiser) black ink, after which they 
were placed in a weak, warmed solution of chromate of sodium for 



440 THE BACTERIOLOGICAL WORLD. 

fifteen minutes. This process was repeated two or three times. Upon 
examination very delicate lines could be seen extending from the ends 
of some of the bacilli, but they were so indistinct that the author 
could not positively affirm that they were flagella. It was this failure 
to satisfactorily stain the flagella of these germs that led Neuhauss to 
employ photography as a means of demonstrating their presence, as 
previously noticed. 

Loeffler's method, i.— Prof. Loeffler ( G ) was the first 
to apply the principle of a mordant in the staining of the 
flagella and cilia of micro-organisms. He succeeded in staining 
the flagella on a large number of motile bacteria by subjecting 
the preparations to the action of a mordant before they were 
brought into the staining fluid. The procedure recommended 
is essentially as follows : 

The Mordant. — To 10 cc. of a 20-per-cent. solution of tannin a 
sufficient quantity of an aqueous solution of the sulphate of iron is added 
to give to the fluid a dark violet color. To this is added 3 to 4 cc. of a 
logwood decoction ( 1 part wood, 8 parts water ). The liquid will now 
have a blackish violet color. Care must be taken not to add an excess 
of the logwood as it would interfere with the staining process. When 
prepared the mordant should be kept in a well-stoppered bottle, and 
in order to preserve it, 4 to 5 cc. of a 5-per-cent. solution of carbolic 
acid may be added. 

The Staining Fluid. — To 100 cc. of a saturated watery solution of 
aniline oil is added 1 cc. of a 1-per-cent. solution of sodium hydrate to 
give to it a slightly alkaline reaction. This alkaline aniline water is 
poured into a flask, in which has been placed 4 to 5 grams of powered 
methylene blue, methyl violet or fuchsin. The flask is vigorously 
shaken and closed with a tightly fitting rubber cork. This solution 
can be kept for a considerable length of time. It must always be 
filtered before using. 

The material to be examined must form a very thin layer upon the 
cover glass. If the germ-containing substance is albuminous, a very 
small quantity of it is added to a drop of sterile distilled water on a 
cover glass and thoroughly mixed with it ; a small quantity of this is 
conveyed to a second cover glass and treated in a like manner ; and 
again from the second a third preparation is made. By this treatment 
the albuminous substance is sufficiently diluted, and the microbes are 
isolated in a watery medium. The preparations are allowed to dry in 
the air, after which the films are fixed by passing the covers, film 
upward, through a flame in the usual manner. 

A few drops of the mordant are poured over the film and the 
cover glass held over a flame until the fluid begins to evaporate. It is 
then removed from the action of the flame, and after a very short time 



THE BACTERIOLOGICAL WORLD. 441 

the mordant is washed off in a stream of distilled water. Care should be 
taken to remove all traces of the -mordant from the edges of the cover 
glass, as it would form, if present, a very troublesome precipitate with 
the staining fluid. The next step is to filter a few drops of the stain- 
ing fluid upon the film. This is allowed to act for a brief time, when 
the cover glass is held over a flame and gently heated. Better results 
are obtained if the staining fluid is only slightly warmed and allowed 
to act for a longer period. As soon as the film becomes darkened ( a 
blackish red if fuchsin is used ) the stain is washed of in distilled water. 
The preparation is now ready for microscopical examination. This 
can be made at once in a drop of distilled water, or the preparation 
allowed to dry and mounted in balsam. 

The microbes with their flagella should be deeply stained, 
resting upon a colorless background if the germs are in a purely 
watery medium, but if albumen is present they are surrounded 
by a uniformly feebly stained medium, the intensity of which 
depends upon the quantity of albumen present. With this pro- 
cess Loefner succeeded in demonstrating the flagella on a large 
number of motile bacilli, spirilla, and upon the motile micro- 
coccus recently described by Ali Cohen. 

Trexeaiaxx's method i. — Soon after the publication of 
Loeffler's method, D. Trenkmann (8) announced his process 
of staining flagella, which in principle is similar to Loeffier's, 
but differs from it in the composition of the mordant and stain- 
ing fluid used, and in several of the lesser important details in 
its application. It is as follows : 

The corer-giass preparations are prepared in the same manner as 
Loefner prepared his. After they have been dried in the air they are 
placed ( without passing them through a flame ) in a fluid composed 
of 1 per cent, tannin and \ per cent, hydrochloric acid and allowed to 
remain in it for from 2 to 12 hours. They are then washed in water and 
transferred to the staining fluid. This consists of a weak solution of 
dahlia ( 2 drops of a concentrated alcoholic solution of dahlia to 20 drops 
of water ). Fuchsin, gentian violet, methylene blue, methyl green, 
Vesuvian, or Victoria blue may be used. The preparations remain in 
the staining fluid for from 1 to -4 hours, when they are rinsed and 
examined. »The flagella are stained with any of the aniline dyes, but 
more satisfactory results are obtained with dahlia, fuchsin or methyl 
violet. The most desirable stain is carbol fuchsin ( 2 drops of concen- 
trated alcoholic solution of fuchsin to 20 drops of a 1-per-cent. solution 
of carbolic acid ) . M 

In a second process recommended in the same article catechu was 
employed. An excess of powered catechu was placed in water and 



442 THE BACTERIOLOGICAL WORLD. 

allowed to macerate for several days and filtered. The filtrate impreg- 
nates the flagella somewhat feeblv without the addition of an acid. 
The cover-glass preparations, prepared as above, are placed in a solu- 
tion composed of four parts of the filtrate and one part of a 5-per-cent. 
solution of carbolic acid. They are allowed to remain in this for from 
2 to 12 hours, after which they are stained in the same manner as by 
the first process. 

A third process is recommended, which consists in placing the 
cover-glass preparations in a concentrated solution of logwood for 
from 2 to 12 hours, after which they are washed and stained in some 
of the aniline dyes. Fuchsin is especially recommended. The addi- 
tion of an acid is said to improve the mordant properties of the log- 
wood solution. The acids proposed are hydrochloric ( ^ per cent.), 
gallic ( £ per cent.), or carbolic ( 1 to 2 per cent.). 

Trenkmann also obtained positive, though less satisfactory, results, 
by three other methods of procedure. First. The preparations are 
transferred from the solution of the extract of logwood to Boehmer's or 
Grenadier's hgematoxylin solution. Second. The specimens are first 
treated in gallic acid and then stained with some of the aniline dyes. 
Third. The preparations are treated first with hgematoxylin solution 
and then with aniline dye. 

By the use of these methods he was able to demonstrate 
the tufts of flagella on many spirilla, especially the spirillum 
undnla and a water bacillus. 

Loeffler's method, ii. — About one year after the pub- 
lication of his first method, Loeffler announced ( 7 ) a second 
process for the straining of flagella which is a marked improve- 
ment over the procedure first recommended. In his careful 
investigation into this subject he succeeding- in discovering an 
important principle by the observance of which it is supposed 
that t..e flagella on all motile bacteria can be stained. This 
consists in the degree of acidity or alkalinity of the mordant. 
In applying this principle he found that those germs which 
were acid-producing organisms (those that will change an 
alkaline medium to an acid one during their development), 
such- as the typhoid bacillus and others, require an alkaline mor- 
dant, and that the alkaline-producing organisms ( those that 
convert an acid into an alkaline medium during their develop- 
ment ), such for example as the comma bacillus, the spirillum 
of Finkler-Prior, the bacillus of blue milk and others, require 
an acid mordant.* The degree of acidity or alkalinity of the 
mordant necessary to secure the staining of the flagella of a 



THE BACTERIOLOGICAL WORLD. 443 

specified germ must be determined by actual experiment ; when 
this is accomplished the method does not differ materially from 
that first proposed. The formulae for the preparation of the 
mordant and staining fluid, together with the details in their 
application, are as follows : 

( 1 ) The Mordant. — To 10 cc. of a 20-per-cent. aqueous solution 
of tannin, 5 cc. of a cold saturated solution of the sulphate of iron and 
1 cc. of an aqueous or alcoholic solution of f uchsin, methyl violet, or 
" Wollschwarzlosung " are added. The fuchsin is especially recom- 
mended. 

The foregoing solution is to be regarded as the standard or stock 
solution to be used, and one which is successfully employed in stain, 
ing the flagella of certain micro-organisms ; but for others the addition 
of an acid or alkali is necessary. Thus for the comma bacillus it is 
necessary to add to the 16 cc. of mordant I to 1 drop of a solution of 
sulphuric acid equivalent to a 1-per-cent. solution of sodium hydrate ; 
for the spirillum rubrum 9 drops of the acid solution are required. For 
the typhoid bacillus 1 cc. of a 1-per-cent. solution of sodium hydrate 
must be added to the 16 cc. of mordant. The bacillus subtilis requires 
28 to 30 drops and the bacillus of malignant oedema 36 to 37 drops of 
the sodium solution. By first determining whether the germ in 
question is an alkali or acid-producing organism the necessary quantity 
of the acid or alkali solution to be added to the mordant can easily be 
determined. 

( 2 ) The Staining Fluid. — The staining fluid here recommended is 
ordinary neutral aniline water in which fuchsin crystals are dissolved 
to saturation. As the aniline water is very nearly neutral a saturated 
solution of fuchsin in it is sufficient. Better results may, possibly, be 
obtained by adding to this as much of a 1 to 100, or still better, 1 to 
1,000 solution of sodium hydrate as it is necessary to bring it almost 
to a point of precipitation. 

Cover-glass preparations should be prepared from a pure culture 
( agar or gelatine ) of the germ to be studied in the manner recom- 
mended in the first process to eliminate all albuminous material- 
Sterilized hydrant water is preferred to distilled water for diluting the 
culture. It is of the utmost importance that the cover glasses should 
be free from all grease or other impurities. In order to clean the cover 
glasses they are boiled in sulphuric acid, washed in distilled water, 
immersed in ammoniated alcohol, from which they are dried on clean 
linen. The film on the cover-glass is fixed by heat, but care must be 
taken not to overheat the preparation. The desired amount of heat can 
be obtained by holding the cover between the thumb and index finger 
over the flame, instead of passing it through the flame by means of 
forceps. By this method overheating is avoided. After heating, the 
film on the cover-glass is covered with the mordant and held over a 



444 THE BACTERIOLOGICAL WORLD. 

flame until steam is given of. It is then removed from the flame and 
after one-half to one minute the cover is rinsed in water, then in abso- 
lute alcohol, and again in water until the mordant is completely 
removed. The film is then covered with a few drops of the staining 
fluid, and the preparation again heated until the solution begins to 
vaporize. It is then removed from the flame, and, after allowing the 
stain to act for about one minute, the cover is washed in a stream of 
water. The preparation can be examined immediately in water, or 
allowed to dry and mounted in balsam. 

By this method Prof. Loeffler demonstrated the flagella on 
a large number of both saprophytic and pathogenic bacteria. 
On some of these tufts of flagella are observed, while others 
possess only one flagellum at each extremity. His paper is 
illustrated by eight photographs of different bacteria studied by 
him, on each of which the flagella can be plainly seen. 

Trenkmann's method, it. — Loeffler J s second process was 
soon followed by Trenkmann's modification of his (Trenkmann's) 
method ( 9 ), in which he introduces iodine water to be applied 
to the preparation immediately after the mordant. This is 
based upon the same principle as the use of the iodine solution 
employed in Gram's method of staining bacteria. This method 
is thought by its author to be superior to the processes previ- 
ously announced, because it is more simple in its details and 
more reliable in its results. The method is as follows : 

( 1) The Mordant. — This consists of a 2-per-cent. solution of tannin, 
to which i to £ per cent, hydrochloric acid is added. 

( 2) Iodine Water. — This is prepared by adding a small quantity of 

pure iodine to a few cubic centimetres of distilled water and allowing 

it to stand, with frequent shaking, for twenty-four hours. The iodine 

solution used in Gram's method, or a drop of the tincture of iodine in 

10 cc. of distilled water, may be used. 

( 3) The Staining Fluid.— This is prepared by adding one drop of 
a saturated alcoholic solution of gentian violet to 10 cc. of distilled 
water. To this, 40 cc. of aniline water are added. The solution 
remains clear. It stains the bacilli and flagella very well, while the 
background is feebly, if at all, colored. 

The cover-glass preparations are prepared with the usual care 
to avoid grease or dirt upon the glass and to secure the necessary dilu- 
tion. Potato infusion is used for such germs as will grow in it. Cul- 
tures in this medium are to be diluted 5 to 10 times ; bouillon cultures 
40 to 50 times, and when the cover-glass preparations are made directly 
from gelatine or agar cultures a dilution of 100 times is necessary. He 
employs boiled water for making the dilutions. A drop of a 10-per- 
cent, solution of alcohol is placed on the preparation and the whole 



THE BACTERIOLOGICAL WORLD. 445 

allowed to dry in the air. The dried preparations, without being 
heated, are placed in the mordant, where they are allowed to remain 
for from 6 to 12 hours or longer. They are then thoroughly rinsed in 
water, after which they are placed in the iodine water, where they 
should remain for about one hour. At the end of that time they are 
again rinsed in water and transferred to the staining solution. This is 
allowed to act for about one-half hour, when the preparations are 
rinsed and mounted for examination, either directly in water or 
allowed to dry and sealed in balsam. With some germs better results 
are obtained by varying the quantity of hydrochloric acid in the mor- 
dant. 

Dowdeswell's method. — In a comparatively recent pub- 
lication Mr. Dowdeswell ( 10 ) states, after referring to the 
method employed by Neuhauss to demonstrate the flagella on 
the comma bacillus, that it is not difficult to detect the flagella 
on the comma forms if appropriate but ordinary means are 
adopted. The optical apparatus required are simply a normal 
retina, a good objective with a moderate angle of aperture, and 
good light. He recommends as a staining fluid an aqueous 
solution of gentian-violet, although other .aniline dyes will 
answer quite as well, and for mounting purposes the acetate of 
potash. There are no special directions given excepting that 
the specimens must be mounted in the fluid mentioned, and not 
in Canada balsam. He also affirms that no difficulty will be 
experienced in staining the flagella on microbes as small as the 
bacterium termo. 

From what has already been said it will be observed that 
the flagella differ to a greater or less degree on the various bac- 
teria on which they have been demonstrated. Dallinger ( 3 ) 
figures the flagella on the bacterium termo as two thread-like 
appendages, one projecting from each end of the rod-shaped 
germ. Their length is much greater than the long diameter of 
the germ, but their thickness one-tenth that of the bacterium. 
He gives the mean average diameter of the flagellum of this 
germ as ¥ o4 1 700 inch, which he determined from fifty meas- 
urements with each of four different lenses. The flagella on 
the spirillum volutans and several other forms are similarly 
figured by the earlier investigators. From the plates and 
descriptions given by Loeffler and Trenkniann it seems that the 
spirilla and a few bacilli possess one or more thread-like fila- 
ments projecting only from the extremities of the organism, 



446 THE BACTERIOLOGICAL WORLD. 

but that the greater number of the motile bacteria on which 
these appendages have been demonstrated are provided with a 
greater or less number of long, delicate, thread-like filaments. 
They appear to be given off either single or in tufts both at the 
extremities and at short and varying intervals along the entire 
length of the rod-shaped organism. In the stained preparation 
the flagella appear either straight, curved or in the form of 
spirals. Many of them are usually detached from the germs, 
presumably during the process of preparation, and appear as 
extremely long, delicate bacilli or spirilla lying between the 
bacteria. This fact renders it difficult to determine with any 
degree of accuracy the number of flagella belonging to any 
particular germ. 

The question may naturally arise, which one of the differ- 
ent methods presented is best adapted for staining the flagella 
on any particular motile germ that maybe under consideration? 
In answer to such a question I would say that Dr. Theobald 
Smith successfully employed Loeffler's second method in staining 
the flagella on both the typhoid bacillus and the bacillus of hog 
cholera. 

I have also found it to give the best results in staining 
the flagella on a considerable number of motile bacteria. 
Although the bacillus of hog cholera is an alkali-producing 
organism, Dr. Smith stained its flagella by using the neutral 
or standard mordant ( 11 ). I have found, however, that the 
flagella on this bacillus stains quite as well by adding from one 
to three drops of the acid solution to the mordant, which shows 
that the range in the reaction of the mordant that can be used 
in staining the flagella, on this germ at least, is much greater 
than might be inferred from the method. This is of consider - 
able importance, as it will, if true with respect to all germs, 
greatly diminish the number of test experiments in order to 
determine the degree of the reaction of the mordant necessary 
to be used with the germ in question. With Trenkmann's 
second process I have been partially successful, but with the 
other methods I have met with only negative results. Thus far, 
however, my experience has been too limited to undervalue 
these methods, or to recommend too highly the one with which 
I have been, in a measure, successful. — Microscojrical Journal. 



THE BACTERIOLOGICAL WORLD. 447 

THE EXAMINATION OF THE BLOOD FOR L AVER AN 'S 
MALARIAL GERM. 

The following, from The Canadian Practitioner of May 
16, 1891, will prove of interest to many "physicians : 

Laveran's directions for examining the blood of malarious 
patients are as follows : The blood should be taken at the 
height of a fever attack, and from a patient who has had no 
quinine for some time. The blood should be taken from a 
finger tip, after a thorough cleansing of the skin to be pierced. 
The cleansing should be such as to prevent all chance of con- 
tamination of the blood as it oozes out. The # drop so obtained 
is to be taken upon a clean cover glass, and a second cover placed 
upon it, so that a thin layer of blood may be obtained between 
two cover glasses. This fresh preparation is then to be exam- 
ined by daylight and with a dry lens of high power. In this 
way one will of tenest be able to see the flagella on the periphery 
of the round, pigmented, free corpuscles. If a dry preparation 
be desired, then the cover glasses must be separated from one 
another and the blood dried by passing the cover glass three 
times through a flame. The specimen can then be examined 
either stained or unstained. Laveran stains with a concentra- 
ted watery solution of methyl blue, before using which he 
washes the cover in a mixture of equal parts of alcohol and 
ether. By this method the nuclei of the white blood corpus- 
cles are stained dark blue, the free, round bodies, or those 
attached to the red blood corpuscles, a pale blue, while the still 
growing corpuscles stain hardly at all. For specimens so pre- 
pared, Laveran recommends dry lenses also. 



448 THE BACTERIOLOGICAL WORLD. 



MISCELLAKEO US. 



THE REALM OF THE MICROBE. 



MRS. PRIESTLY. 

Nineteenth Century, London, May. 

All living tilings throughout the animal and vegetable 
kingdoms are composed of cells, springing, in the first 
instance, from one single cell. The entire human child, with 
all its great future possibilities, is, as Haeckle says, in its first 
stage only a single ball of protoplasm monerula. But at the 
other end of the scale we have microscopic organisms, existing 
as simple cells, capable of performing all the functions of life, 
and of playing an important role in the economy of nature. 

Pasteur, in the course of his study of the germ theory, 
argued that all that has lived must die, and all that is dead 
must be disintegrated, dissolved or gasified ; the elements 
which are the substratum of life must enter into new cycles of 
life. One grand phenomenon he saw, presides over this work 
— the phenomenon of fermentation. 

What, then, is the cause of fermentation ? In order to 
answer this profound question, Pasteur devoted himself to the 
study of the microscopic beings, which he finally divided into 
two great classes, the aerobies and the anaerobies, those which 
require free oxygen for their existence, and those which are 
killed by the presence of free oxygen, although able to wrest 
oxygen from the materials whence they derive their nourish- 
ment. 

The aerobies are those which begin work on the surface of 
things, their mission being to clear the earth, by a process of 
slow combustion of all that is dead. The anaerobies, working 
simultaneously, spring into activity underneath the surface of 
putrescible matter, and, dying on exposure to the free oxygen 
of the air, are, in their turn, swept away by the aerobies on the 
surface. 



THE BACTERIOLOGICAL WORLD. 449 

Thus the two great classes of minute living organisms 
co-operate towards the fulfilment of a common end, the one 
beginning work which the other takes up and completes. Bat 
for their united efforts we should cease to live, for the earth would 
be littered with fallen debris and the organic matter of every 
kind, all of which it is their function to transmute into the very 
elements which are necessary to life again. In the parts of 
the earth where these organisms do not exist there is no vege- 
tation, no organic matter, no life of any kind ; the region is 
one vast field of ice, a sandy desert, or an expanse of eternal 
Snow. When perchance these desolate places are invaded by 
living creatures who starve and fall by the way, there is no 
decay, for the organisms whose office is that of putrefaction 
are not present to perform their analytical functions. si Thus," 
says Pasteur. 

" The destruction of everything that has lived reduces itself to the 
simultaneous action of these three great natural phenomena, fermen- 
tation, putrefaction and slow combustion. The carbon, the hydrogen, 
the nitrogen of organic matters are transformed by the oxygen of the 
air, and by the action of these aerobies, into carbonic acid, vapor of 
water, and ammonia gas." 

Having thus recognized the yast importance of these 
minute organisms, Pasteur, the chemist, watched, with unceas- 
ing interest, the work of these greater chemists in Nature's 
own laboratory. Not only were they engaged in the immense 
business of preserving the balance between life and death, but 
they participated largely in the everyday work of the world, 
and were taking an active part in the industries of man. The 
power of these lowly and invisible creatures is so enormous that 
they are well called " the masters of the world," If we neglect 
the laws of health they are the Nemesis which deals punish- 
ment ; for in the steady, ceaseless pursuit of duties, they spare 
none. All organic matter is the same to them whether it be 
the human body living or the dungheap at the cottage door. 

In nature, some of these organisms live only in living 
bodies, arid are consequently distinguished as parasites, while 
others live on dead bodies, and are known technically as sapro- 
phytes, but in the laboratory both forms as a rule can be culti- 
vated on artificial material, rabies being an exception, as it can 
only be cultivated in the living bodies of animals. The Kev. 



450 THE BACTERIOLOGICAL WORLD. 

Dr. Dallinger, who keeps a museum of living putrefactive 
organisms, describes one of them, to which he has devoted 
special attention, and thus reveals the marvelous workings of 
nature in a world we cannot see. This creature is so minute 
that the average measurement is T o j, 00 of an inch in length 
and 1 9 \ of an inch in breadth. Nevertheless, it has six 
flagella, each one of which is three times as long as the length 
of its body. The movement of these creatures is exceedingly 
beautiful and graceful, with the long flagella waving to and fro. 
After a moment's rest the group under observation will be seen 
to start into active movements in a series of wavelike leaps, 
reminding one of the movements of a shoal of porpoises, which 
is continued for from ten to fifteen undulations. This organ- 
ism, unlike others described by Dr. Dallinger, was never 
anchored and had no power of attaching itself, but by freely 
darting upon the matter attacked, such matter was visibly, in 
the course of half an hour, reduced in size and altered in shape. 
Each one in turn, in a space of about two seconds, comes into 
contact with the particle and at once recedes to a distance of 
four or five times the length of the flagella, and instantly again 
darts upon the object, and this may be continued by given 
forms for hours. 

Fifty to a hundred may be seen with ease in one micro- 
scopic field pursuing their untiring work. It is the more 
entrancing that it is apparently rhythmical, not like the meas- 
ured march of a regiment, but the rhythmic movement of a 
peal of bells. 

These are examples of the putrefactive organisms at work 
throughout nature, in company with a vast number of other 
forms, all engaged in ridding the world of putrescible matter. 

It is difficult to imagine any study more fruitful in happy 
results to mankind or more intensely fascinating to the student, 
than that comprised in the new world of the invisible. 

The point where perhaps most discussion has taken place 
is that on the border line between the pathogenic and -the non- 
pathogenic organisms. In some cases, for instance, that of the 
bacillus subtihs and the bacillus anthracis the difference is so 
slight that it can hardly be detected under the microscope, yet 
the one is harmless to man, and the other deadly. 



THE BACTERIOLOGICAL WORLD. 451 

From the study of the familiar fermentation of yeast, 
Pasteur passed to the study of the diseases of beer and wine, 
and thence to the silk-worm disease, which latter proved the 
connecting link which established the general relationship 
between the diseases of beer, wine, and all putrescible things, 
with the diseases of all living creatures from silk- worms to 
man, — The Literary Digest. 



The medico-legal value of the gonococcus. — Since 
the discovery of the gonococcus, and since its general accept- 
ance by the medical profession as a means of differentiation 
between specific and non-specific blennorrhea, hopes have 
been entertained that it might prove of value in medico-legal 
cases, especially in those instances of vulvo- vaginitis occurring 
in children, and often giving rise to suspicions of criminal prac- 
tice. Neisser, Lober and Kratter all assert very positively that, 
by the aid of the microscope, they can determine without 
question whether the vaginal discharge is gonorrheal, depend- 
ent upon impure contact, or whether it is non-specific, in 
which case it may be evoked by any one of a large number of 
irritants. 

Vibert and Bordas ( La Medicine Moderne, November 13, 
1890) point out that to be of value in legal medicine the proof 
must be absolute, and that probabilities are of little avail. The 
vagina contains, even in health, a vast number of micro- 
organisms. If the secretion of an acute vaginitis occurring in 
the person of a little girl is examined, diplococci, presenting all 
the characteristics of the gonococcus, are usually observed. 

In those cases examined by the authors, the -cause of 
inflammation was purely traumatic, and, according to the evi- 
dence, it was scarcely possible that gonorrheal pus had come 
into contact with the mucous membranes. The discharge of 
these cases contained diplococci resembling in every respect 
gonococci. From four cases pure cultures of this micro- 
organism were obtained, in the remainder there were other 
varieties. The diplococci were found always in the interior of 
the pus-corpuscles, and were very abundant. In form, in 
dimensions, in position, in grouping, in habitat, and in reaction 



452 THE BACTERIOLOGICAL WORLD. 

to staining fluids, these micro-organisms were identical with 
gonococci. 

It is well known that in the Gram method, the aniline 
stain affects all micro-organisms likely to be found in blennor- 
rhoeal pus, excepting the gonococci ; hence, when a preparation 
previously colored is washed for some minutes in Gram's solu- 
tion, and is then dehydrated in alcohol, the gonococci are 
completely decolorized. The diplococci of non-gonorrhoeal pus 
act in the same manner. 

Other authors, and especially Bumm, have asserted that 
there are micro organisms which cannot be distinguished either 
morphologically or by staining methods from the gonococcus. 

It would seem, from the present evidence, that the results 
of microscopical examination of vulvo-vaginal discharges are 
not sufficiently conclusive to warrant a positive statement either 
for or against impure contact. — Philadelphia Medical News. 

Resorcin in septic poisoning. — Andeer (Chem. Centra/- 
Matt), in view of the many deaths which have recently 
occurred from dissection wounds, poisoning by cadaverous 
alkaloids, etc., calls attention to the uniform efficacy of resorcin 
as a remedy in such cases. On the other hand, in antidotmg 
the venom of serpents and insects, it is only occasionally suc- 
cessful — a fact which the author explains by supposing that 
resorcin acts antiseptically upon morbid matters which are 
alkaline or neutral in reaction, but is powerless or hurtful when 
they are acid. — Drtiggists' Circular. 

Fish poison. — The Vierteljahrsschrift fur Gerichtliche 
Medicin calls attention to a very important matter in connection 
with th@ use of fish as a food. It appears that an investigation 
of the result of eating fish, preserved on ice for use in the Lon- 
don markets, has led to the discovery that those were most 
dangerous which were kept in immediate contact with the ice. 
Poisoning by fish which had not been in contact with ice was 
not observed at all. This is attributed to the influence of the 
water derived from the ice, and bearing whatever impurities it 
has had before being frozen, which promotes the formation of 
the animal alkaloids known as fish poison. — Druggists' Cir- 
cular. 



THE BACTERIOLOGICAL WORLD. 453 

Prevention of diphtheria.— Dr. Augustus Oaille con- 
cludes a paper on this subject, read before the American 
Pediatric Society, by saying that at the present time his convic- 
tion is firm that, in the absence of filthy carious teeth and nasal 
obstruction from adenoid vegetations or greatly enlarged ton- 
sils/ the daily prolonged use of mild antiseptic liquids by means 
of spray, insufflation or gargling, will prevent diphtheritic 
infection. He claims, also, that this procedure is indicated for 
those who aie exposed to diphtheritic infection; and, also, as^ 
routine treatment for every case of nasal catarrh, pertussis, 
measles and scarlet fever, in fact, for every condition in child- 
ren in whom the naso-pharyngeal mucous membrane is hyper- 
emic or congested, and therefore predisposed to diphtheritic 
infection. — Druggists' Circular. 

Salt in milk for children. — The addition of sodium 
chloride prevents the solid coagulation of milk by either rennet 
or gastric juice. The cow's milk ought never to be given with- 
out table salt, and the latter ought to be added to a woman's 
milk when it behaves like cow's milk, in regard to solid curd- 
ling and consequent indigestibility. Habitual constipation of 
children is influenced beneficially, since not only is the food 
made more digestible, but the alimentary secretions, both serous 
and glanular, are made more effective by its presence. — Arch, 
of Ped. 



READING NOTICE. 



A solid train to Buffalo, N. Y., leaves St. Louis 8 :05 A. M. daily 
via Big Four Route, with through sleeping car to Grand Central Sta- 
tion, New York City, via Lake Shore & New York Central Ry. This 
is the Famous " Southwestern Limited," vestibuled from end to end ; 
Cafe & Dining Car service. For time tables, rates, etc. , apply Big 
Four Ticket Offices, Cor. Broadway & Chestnut Sts. and Union Depot, 
St. Louis, Mo. 

As usual, the Big Four Route will make low excursions rates on 
account of the 4th of July ! Tickets will be on sale July 3d and 4th 
between all local points within a radius of 200 miles from initial point 
of ticket at one fare for the round trip. Tickets good going on date of 
sale and good returning to and including July 6th, 1891. Turn out, 
one and all and celebrate the glorious fourth. For tickets and full 
information call on or address. W. F. Snyder, 

Gen. Western Agent, C. C. C. & St. L. Ry., St. Louis. 
D. B. Martin, 
General Passenger Agent, Cincinnati, O. 



454 THE BACTERIOLOGICAL WORLD. 

LITERATURE. 

HARPER'S MAGAZINE. 

CONTENTS FOR JULY. 

An Imperative Duty — A Story Wm. Dean Howells. 

Peter Ibbetson — A Novel George Du Maurier. 

Oliver Wendell Holmes — A biographical 

and Critical essay George Win. Curtis. 

The Avon from Evesham to Tewkesbury. .A. T. Quiller Couch. 

London — Saxon & Norman Walter Besant. 

The Episode of the Marques de Valdeflores . Thos. A. Janvier. 

American Riders Frederic Remington. 

The Republic of Paraguay Theodore Child. 

Briticisms and Americanisms Brander Matthews. 

Christianity and Socialism Rev. James M. Buckley. 

Literary Notes Lawrence Hutton . 

THE POPULAR SCIENCE MONTHLY. 

CONTENTS FOR JULY, 1891. 

The Development of American Industries 
since Columbus. VI. The Revolution 
of Wool Spinning and Weaving. 
( Illustrated.) S. N. Dexter North. 

Man and the Glacial Period. (Illustrated.). Prof. G.Frederick Wright. 

Sanitary Improvement in New York dur- 
ing the last Quarter of a Century General Emmons Clark. 

Deportment of Savage Negroes Paul Reichard. 

Pollen : Its Development and Use. (Illus- 
trated.) Joseph F. James, M. Sc. 

The Meteoritic Hypothesis J. Ellard Gore, F. R. A. S. 

Our Agricultural Experiment Stations Prof. C. L. Parsons. 

A Coming Solution of the Currency Ques- 
tion Charles S. Ashley. 

Scientific Dreams of the Past Albert de Rochas. 

The Colors of Letters David Starr Jordan. 

Animal and Plant Lore. IV. ( Concluded. ). Mrs. Fanny D. Bergen. 

Hoff ding's Outlines of Psychology 

The Quianganes of Luzon Prof. F. Blumentritt. 

On the Wings of the Wind 

Sketch of George Catlin. ( With Portrait.) 

Editor's Table The New Jesuitism and Social Reform. — Charity as a 
Fetich 

Literary Notices 

Popular Miscellany - 

Notes 



THE BACTERIOLOGICAL WORLD. 455 

LIPPINCOTT'S MONTHLY MAGAZINE. 

CONTENTS FOR JUNE. 

Gold of Pleasure George Parsons Lathrop. 

Some Familiar Letters by Horace Greeley. 

IV . . Edited by Joel Benton. 

A Literary Pet ,. Edgar Fa wcett. 

Alexandra, Princess of Wales t Lucy C. Lillie. 

Oracles Clinton Scollard. 

A By- Way in Fiction Agnes Repplier. 

Is Alaska worth Visiting ? Grace Peckham, M. D. 

Horace, Ode IV. , To Sestius Caroline Augusta Furness. 

In the Thorvaldsen Museum C. H. Herford. 

Beneath the Trees Charles Morris. 

Sonny Mary E. Wilkins. 

The College Settlement Hester Dorsey Richardson. 

Before the Hour Florence Earle Coats. 

Two Recent Novels : 

"Atnian" A. H. W. 

" Romance of a Spanish Nun " Frederic M. Bird. 

With the Wits. ( Illustrated by leading artists.) 



LISTERINE 



Antiseptic I C J I f\ I NoN - ToxIC 

Prophylactic . ^^^ r— I < I \ I p- Non-Irritant. 

Deodorant. I V J 1 I I \l Non-Escharotic 



FORMULA.- Listerine is the essential antiseptic constituent of Thyme, Eucalyptus, 
Baptisia, Gaultheria and Mentha Arvensis, in combination. Each fluid drachm 
also contains two grains of refined and purified Benzo-boracic Acid. 
DOSE.— Internally : One teaspoonful three or more times a day ( as indicated ) 
either full strength or diluted, as necessary for varied conditions. 
LTSTEBJNE is a well-proven antiseptic agent — an antizymotic— especially useful 
in the management of catarrhal conditions of the mucous membrane ; adapted to 
internal use, and to make and maintain surgical cleanliness— asepsis— in the treat- 
ment of all parts of the human body, whether by spray, irrigation, atomization or 
simple local application, and therefore characterized by its particular adaptability to 
the field of 

PREVENTIVE MEDICINE -INDIVIDUAL PROPHYLAXIS. 



* LISTBRIUE * 

Destroys promptly all odors'emanating from diseased gums and teeth, .aid will be 
found of great value when taken internally, in teaspoonful doses, to control the fer- 
mentative eructations of dyspepsia, and to disinfect the mouth, throat and stomach, 
It is a perfect tooth and mouth wash, indispensable for the dental toilet. 
Descriptive literature upon request. 



LAMBERT PHAEMACAL CO., ST. LOUIS, MO. 

AGENCIES: 

S MAW SON & THOMPSON, ROBERTS & CO., S. PAPPENHEIM, VILANOVA HOS. Y CIA. 

LONDON, E C. PARIS. BERLIN, W. BARCELONA. 



CX/CTIB RATES 5 



m 




u 



L 



J 




( ILLUSTRATED.) 
$3.00 PER YEAR IN ADVANCE. 



We offer it with the following publications at the rate indicated : 

The Journal I American Medical Association 

( WEEKLY.) 
Price, $5.00. With Bacteriological World, $7.00. 



The Journal of Comparative Medicine and Veterinary Archives. 

EDITORS : 

W. A. Conklin, Ph. D., D. V. S., Director Zoological Gardens, N. Y. 

Rush S. Huidekoper, M. D., Veterinarian, Prof. American Veterinary 

College, New York. 
Price, $3.00. With Bacteriological World, $5.00. 



THE fM\U\ VETERINARY REVIEW. 

Editor in Chief: Prof. A. Liautard, Principal, American Veterinary 

College, New York. 
Price, $3.00. With Bacteriological World, $5.00. 



THE CINCINNATI MEDICAL NEWS, 

Editor : J. A. Thacker, A. M., M. D., Cincinnati, Ohio. 
Price, $2.00. With Bacteriological World, $4.50. 



THE MEDICAL HERALD. 

Editor : Daniel Morton, M. D., St. Joseph, Mo. 

Price, $2.00. With Bacteriological World, $4.50. 



ST. LOUIS CLINIQUE. 

Editors : Dr. William Porter and a Corps of Associates, 

St. Louis, Mo. 

Price, $1.00. With Bacteriological World, $3.75. 



Address THE BACTERIOLOGICAL WORLD, Columbia, Mo. 

Or for quicker attention: DR. T. J. TURNER, Mexico, Mo., 

Who has full authority to attend to all business matters for the 
Bacteriological World Company. 

* For the United States, Canada and Mexico only. 



m 



F 



-< 



A^TT? DTOT QP TP A 
Ji JjUlULUulun 




A Monthly Illustrated Magazine Devoted to the Study of Micro-Organisms and Specific Maladies. 



Original articles, clinical reports, books for review, exchanges, scientific 
correspondence, should be addressed to the Editor of the Bacteriological World, 
Columbia, Missouri. 

Subscriptions, advertisements, and all business matters are attended to at the 
same office. 

Authors of original articles published in this magazine will be entitled to twenty 
copies of the number containing it, provided, that they request the same in writing 
when sending the communication. Reprints at actual cost. 

Except the three months' trial subscription, at75cts., no subscription will be 
received for less than one year, commencing in January. 

Discontinuation :— At the end of the year we shall consider subscribers indi- 
vidually as wishing the magazine continued, unless they order it stopped in writing 
before the first of next January. 



collaborators: 



Paul Gibier, M. D., New York. 
Prof. W. T. Belfield, M. D., Chicago. 
Prof. A. W. McAlester, A. M., M. D., 

Missouri University Med. Dep't. 
Prof. Henry P. Loomis, M. D., 

New York . 



Prof. L. Bremer. M. B\, St. Louis, Mo. 
J. W. Stickler, M. D., Orange, N. J. 
Prof. Paul Schweitzer, Ph. D., 

Missouri University. 
Prof. T. J. Burrill, Ph. D., 

Illinois University. 



B. T. Galloway, B. S., Section of Vegetable Pathology, Washington, D. C 



U. S. A., JULY, 1891. 



7. 



EDITORIAL. 



THE ROLE OF MICROBES IN THE DIGESTION OF FOOD. 

It was Pasteur who first ventured the opinion that possibly 
microbes were useful factors in normal digestion in the animal 
world. Duclaux, in his celebrated biological chemistry, 
impressed one as believing, to a limited degree, in this theory. 
We are told,. though we confess never having read it, that 
Vaughan is imbued with the same idea also, in a limited sense. 

Judging from natural laws governing nourishment imme- 
diately after birth in large animals, one would think that this 
must be a somewhat unreasonable theory. Indeed, it cannot be 
shown that, in case of a child excluded from bacteria from the 
moment of its birth, life would cease, because digestion must 
fail in the absence of digestive ferments in the alimentary tract. 



458 THE BACTERIOLOGICAL WORLD. 

In fact, the digestion of the first milk in the young, taken 
from the breast, is free from microbes and goes into a canal 
likewise unpolluted ; and there is no good reason to believe 
that if these conditions were kept up until the adult stage the 
subject must starve. An organism has naturally the powers 
and activities necessary for digestion and other functions, irre- 
spective of the aid of other foreign living organisms as a 
necessary adjunct. 

True, there are to be found in the mouth and alimentary 
tract bacteria capable of causing certain changes comparable to 
the natural digestion. For instance, starch may be converted 
into sugar in the mouth ; it is possible even that ptyalin is not 
so active in that sense as was believed, and that microbes 
increase this converting power. But this does not prove their 
necessity in the mouth, for the animal is capable of operating 
this transformation in the small intestines by the action of 
certain principles in the pancreatic juice. 

It is a fact that microbes produce, for the purpose of their 
own nourishment, digestion of food similar to digestion by 
complex beings, but it is for the sustenance of their own 
structure and not directly of other beings. Their action in this 
line in the alimentary canal is purely accidental, in the sense 
that it is not a necessary phenomeuon of the growth and life of 
beings thus harboring them. 

But what role do these microscopic organisms — minute 
ferment germs — play in cases of indigestion ? Here they are, 
indeed, a great factor, and, if they may be valuable in acci- 
dentally assisting in the dissolution of too much food ingested, 
or a quantity of foodstuff that the animal organism cannot fully 
overcome, they may also cause, during the process of trans- 
formation, very serious disturbances. 

The stomach of a human being for instance, like the heart 
and other organs, has a limited capacity beyond which it cannot 
safely be taxed. There is a reserve force for a case of emer- 
gency, say, to digest the pound or more of surplus albumenoids 
and other stuff ingested by the bon vivant or glutton at a four- 
hour-long banquet, just as the heart may beat one hundred 
and eighty and more a minute in the efforts to carry oxygen to 
the tissues in case of disease, fever, but this is an unnatural 



THE BACTERIOLOGICAL WORLD. 4 59 

activity which, if continued, must sooner or later result in 
incapacitating the stomach ; it eventually becomes exhausted 
and fails to secrete the necessary digestive fluids in sufficient 
quantity and proper quality. The same thing may be said of 
all the other overtaxed digestive organs. If they are kept 
under constant strain, secreting, churning to their utmost 
capacity, they must soon fail, as the street-car mule, pulling to 
its utmost, drawing daily on its reserve force under the driver's 
whip, is bound to fail. Exhaifstion, more or less complete, is 
inevitable. What then follows ? 

The gastric juice, being deficient, has lost partly its germi- 
cidal properties as well as digestive properties. The food 
swallowed fails of complete digestion ; that which is digested 
is acted upon simultaneously by the natural ferments of the 
body and by the ferment (or decomposition or transformation) 
microbes ; the surplus that fails entirely of natural digestion is 
decomposed largely, if not entirely, by microbes. 

What is the result of such complicated and unnatural 
transformations ? The result is practically as we would find 
them had we followed the process in a tin dish. The sum total 
is putrefaction in part at least with the usual results. It is not 
a natural process of digestion, but a " rotting process," if you 
will pardon the expression, which though it does aid in freeing 
the body from its inconvenient load does so in a very dangerous 
and often a very painful way, thereby complicating the func- 
tional troubles brought about by unwise feeding, and doubtless 
often producing some organic lesions that eventually end in 
death. Consider the results of the decomposition of foods 
serving as man's nourishment for instance, and see what prod- 
ucts may be the result of chronic or acute indigestion under the 
converting influence of bacteria. In the phenomena which 
follow, " sugar is transformed into lactic acid, mannite, dextrine, 
glycerine, butyric acid,, mucilage, etc Alcohol is transformed 
into acetic acid, urea into carbonate of ammonia, etc. There is 
generated the putrid poisonous substance named sepsin ; the 
septic alkaloids of Zulzer and Sonnenschein ; the ptomaines 
found by Selim, G-autheir, Breiger, Vaughan* ; certain nar- 



* Name added to quotation by editor. 



460 . THE BACTERIOLOGICAL WORLD. 

cotics, leucine and tyrosine, butyric, palmitic, margaric and 
fatty acids ; volatile products, iodol, phenol, scatol, sulphurated 
hydrogen, ammonia, carbonic acid, water," etc. 

With such a quantity of chemical products ( many of which 
are pernicious) produced in our bodies, is it a wonder that the 
head aches, and the whole anatomy of the dyspeptic is sore, and 
he sometimes ends in the insane asylum ? Is it not rational to 
conclude that when one converts one's stomach into a labora- 
tory for generation of products of decomposition, putrefaction, 
they produce disease by their direct local action, their action 
on the nervous centers, their action on the blood and every cell 
of every organ of the body when absorbed and carried to them 
with the food intended to nourish them ? Verily, indigestion is 
a method of poisoning by nature's processes. Indeed, when 
produced by gourmandizing, intemperance, knowing both to be 
injurious, it is a species of slow suicide ; leaving aside the 
cause and intent, it differs from the arsenic poisoning mainly 
in the degree of rapidity in accomplishing the end. 



THE BACTERIOLOGICAL WORLD. 461 



LESSONS IN BACTERIOLOGY. 



LESSON SIXTH {Continued). 



BY PAUL PAQUIN, M. D. 



CULTURE MEDIA, ETC. 

Solid cultures ( continued ). — All broths which have 
been gelatinized or to which has been added agar-agar for the 
purpose of solidifying should be put in test tubes or small cul- 
ture flasks, vials, and the like, while still warm ; for, in cooling, 
they will lose their fluidity and become a jelly-like mass. If 
one desires to keep such material in stock it should be placed 
in a large flask ( previously sterilized, of course, just as the test 
tubes, etc, ), and then the final sterilization process carried on. 
When any of the material is needed, put the flask in a water 
bath, and, as soon as melted, fill the culture recipients as may 
be desired. Always remember that whenever a stock flask is 
opened it should be sterilized again at once, for air germs are 
likely to have entered it and may soon grow. 

To make any broth clear, transparent and free from cloud- 
iness it is better to use the albumen of egg in the follow- 
ing manner : For each pint of culture broth beat the white 
of one egg, in one, two or three ounces of water ; let the broth 
cool a little after the boiling and before last filtration, then add 
the beaten egg. Boil again a very minutes in the water bath 
and filter. I have mentioned the addition of egg albumen 
before but did not state the mode of procedure. 

To obtain a larger surface in the tubes, allow them to cool 
in a slanting position after sterilization. 

Blood serum. — This material is sterilized properly only 
with great difficulty. It is indeed almost impossible to succeed 
in getting a pure, good transparent serum medium without 
special apparatus. The reason will be apparent to all. Steril- 
ization is usually effected only by very high heat applied once, 



462 THE BACTERIOLOGICAL WORLD. 

or repeated heating at nearly or' quite 212° F. In either case 
blood serum coagulates at such temperatures as the albumen of 
an egg in boiling water and becomes white, opaque ; its surface 
soon dries and it is unfit for the proper cultivation of bacteria. 
The usual way of getting blood serum for culture media fs 
as follows : A glass jar, say a Mason fruit jar, plugged with 
cotton is wrapped thoroughly in a few thicknesses of dense 
cotton cloth, linen or paper, and then sterilized at dry heat in 
the manner indicated for such sterilization. The jar is carried 
to a slaughterhouse and the wrapping removed only just before 
filling it. Remove plug of cotton carefully, give it to an aid to 
hold, with under surface downwards, then with the greatest pos- 
sible care and cleanliness allow blood from a calf (or older 
bovine ) to enter the jar. When nearly full replace plug of cotton 
and carry the blood to your work room (laboratory) without 
shaking it if possible. Once there, place the jar in a dish of 
cold water and put the whole in a cool place ( ice box ) to 
allow the clot of solid material to form, and the serum to separate 
all around it. After about thirty hours the serum may be drawn 
carefully from around the clot (without disturbing the latter ) 
by means of a glass pipette sterilized or a sterilized rubber 
syphon and made to run into sterilized culture tubes. All care 
possible should be taken to transfer the serum from the jar to 
the culture flasks or tubes, because air germs might enter with 
serious consequences, owing to the fact that blood sterilization 
must be effected at a comparatively very low temperature. Do not 
moisten the lips or the upper portion of the inside walls of the 
tubes ; because, in the heating to follow, they may fail of steril- 
ization*if the water bath process is used, and this is the only 
way available where one has not gone to the expense of fitting 
up a laboratory with an assortment of more or less expensive 
instruments. The test tubes ( always sterilized ) being filled 
and plugged, place them in a wire basket or perforated tin pail 
and plunge the whole into a water bath kept at a temperature 
of about 120° F., which may be maintained thus by close watch- 
ing by means of a low gas jet or an alcohol lamp. Repeat this 
heating once a day for about a week when sterilization ought to 
be complete and the medium sufficiently clear to serve for the 
cultivation of bacteria. In order to get a larger culture surface 



THE BACTERIOLOGICAL WORLD. 463 

of this solid material it is only necessary to incline the tubes 
with care within the wire basket or pail, if the size of it will 
admit, and to leave them untouched until the last heating. 

Dry-heat sterilization of blood serum is effected by means 
of a special serum " coagulator and sterilizer," which requires 
no special technology to manage. 

The heating is repeated at dry regulated heat as for the 
method just described. Certain instruments are on the market 
to regulate the heat of such apparatus as well as to regulate the 
heat of culture ovens or incubators to be designated later. 
They are very handy, but not indispensable. 

Potato culture. — The potato constitutes an excellent 
nutrient soil for the cultivation of bacteria of various kinds, 
though it does not afford the growth of all germs. The field 
of culture is prepared by sterilizing mashed potato on glass 
plates about six inches by four, in shallow special dishes or 
round trays covered by another larger but similar vessel serving 
as lid. 

The -usual potato cultures, however, consist merely of 
cooked potatoes cut in two under the most carefully aseptic 
methods — the two cut surfaces serving as the culture field. 

Take neat medium-sized potatoes having no injuries, few 
eyes, or specks ; wash several times in clean water rubbing them 
with a brush or stiff rough towel and rinsing in clean water 
( scalding preferred ) ; then immerse them for an hour in a 
solution of bichloride of mercury 1-1000. Place them in a 
perforated pail or wire basket and cook them by steam in an 
ordinary covered kitchen steamer or in a special laboratory 
steamer. In thirty or forty minutes the steaming will have 
destroyed the remaining external micro-organisms and cooked 
the potato sufficiently. Let them cool down without opening 
the steamer and they are ready to be cut in two and serve as 
culture material. 

Before doing this, however, another step must have been 
taken in another direction. Whilst the potatoes were cooking, 
dishes to receive them as culture media should have been 
prepared. These dishes consist of circular trays in pairs in the 
form of a stiff straw hat without rim — the larger forming a lid 
for the smaller, just as if in covering low tumbler (glass) with 



464 THE BACTERIOLOGICAL WORLD. 

a larger one fitting loosely over the mouth. These dishes are 
washed clean, sterilized by shaking a 1-1000 solution of 
bichloride of mercury in the dishes closed, allowing the 
solution to run out gradually by tipping over and without 
removing the cover entirely. The object is to wash these 
dishes thoroughly with bichloride . ( after using only water ) ; 
drain them and yet do the whole in such a way as to expose 
the inside surfaces as little as possible to the air. In order to 
keep dampness in the potato chamber thus prepared a ( circu- 
lar ) piece of filter ( or other ) paper is saturated with the 
bichloride solution and placed at the bottom. It is safer, 
however, to put this paper in in washing ; put a good deal of the 
liquid in and allow it to come in contact with every portion of 
the dishes inside without exposing them any further to the air. 
Then dip the hands in the same kind of solution ( kept in another 
dish for the purpose ) ; sterilize the blade of a sharp enough 
kitchen knife by passing it in the blaze of a gas burner or of an 
alcohol lamp three or four times ; pick a potato from within 
the steamer with the index finger and thumb of the left hand, 
the lid having been removed most carefully by the right hand, 
holding also the knife with edge of blade upward, or better 
by an assistant ; sever the potato in two and carefully place in 
the dishes or circular glass trays. 

The cut surfaces, at this stage, are still 'in contact sticking 
together; by carefully lifting the lid to one side to introduce the 
knife blade they can be separated with the ( freshly sterilized ) 
point forced into the incision, thus turning upward the cut 
surfaces of each half of the potato. 

Leave the potatoes thus prepared two or three days at a 
warm temperature before inoculating them with any bacteria ; 
the object is to allow the growth of any air germ that may have 
accidentally fallen on the potato during the various operations. 
If any growth should develop from air germs the media is 
useless for any inoculation or special growth. 

During all this work one thing must be kept constantly in 
mind, i. e., the object sought is two clean cut surfaces, perfectly 
sterilized in the center of a sterile potato, inclosed in sterile 
dishes. A little practice will soon render one perfectly com- 
petent to accomplish this easily. It is unwise to try and 



THE BACTERIOLOGICAL WORLD. 465 

cultivate special bacteria on a potato surface, or any other 

culture media, before giving sufficient time to the material to 

develop any growth of germs fallen accidentally therein or 

thereon. 

(Seventh lesson in August number.) 



ORIGINAL ARTICLES. 



TRANSFORMATION AND ELIMINATION OF THE ORGANIC 
MATTER BY THE PYOCYANIC BACILLUS. 



BY MESSRS. A. ARIs"AUD AND A. CHARRIK. 
Before the Academy of Sciences, Paris, France. 

In a preceding note, we have made known the transfor- 
mations resulting in the organic matter (asparagine ) under the 
vital influence of the pyocyanic bacillus. We have studied the 
mode of elimination of nitrogen ; it remains, among other 
things, to establish the carbone of the carbon of this asparagine. 

The weight of the carbon of the liquid of culture is, at 
the start, 1 gr. 600 per litre, quantity corresponding to the 5 
grams of the asparagine employed. The direct dosing of 
CO 2 disengaged, during the fifteen days of the evolution of the 
microbe, has given some figures coming close, in average, to 
those that it is possible to calculate, in holding account of the 
carbon fixed in the microbian protoplasma, and of that which 
enters in the composition of the substances of secretions, sub- 
stances not yet determined, constituting the fixed residue. The 
results of the analyses are as follows : 

RESPECTIVE QUANTITIES OF SUBSTANCES ELABORATED AT THE COST OP 
THE ORGANIC MATTER (ELIMINATION OF THE CARBON). 

Total carbon in 1 litre of culture "1 gr., 600. 

Carbon released in the state of carbonic acid.l gr., 160 or 72.5 per 100. 
Carbon combined in the microbic proto- 
plasma 0.221 mgr. or 13.8 per 100. 



466 THE BACTERIOLOGICAL WORLD. 

Carbon of the soluble substances, fixed 

products of secretions not determined(l). 0.216 mgr. or 13.5 per 100. 

Carbon in combination secondary prod- 
ucts, pyocyanine, methylamine and loss 
in the analysis 0.003 mgr. 

*Gr. stands for grams not grains . 

The determination of the oxygen fixed by the bacillus 
presents interest. Our experiments have not, until now, 
given absolutely concordant results. In order to arrive at 
exact results, we have undertaken some analyses by different 
methods. Still the analysis, in accord with the calculation, 
shows that this quantity of absorbed oxygen is in relation to 
the proportion of O 2 which has been created ; it can be esti- 
mated, at from one and a half to two times the volume of the 
culture. 

In the void the evolution is poor ; CO 2 permits no develop- 
ment. In hydrogen we note fairly great activity ; ammonia is 
formed, we have not yet studied by which mechanism. 

We have completed our researches in making the medium 
vary. Instead of utilizing the asparagine, we have taken the 
gelatine, more complex in constitution, taking care to add 
the necessary nutritive salts. The curve of elimination of the 
nitrogen matter is then more regular than the asparagine ; 
it seems that the diastase of the bacillus does not intervene 
and that this bacillus assimilates directly the elements of the 
gelatine. The results are resumed in the following table : 

Grams. 

Total nitrogen at the start of the culture, March 30, 1891 0,757 

Nitrogen eliminated in the state of ammonia, 90 hours after the 

start 0.1344 

April 7, 196 hours after the start 0.3444 

April 9, 234 hours after the start 0.4564 

April 14, 360 hours after the start 0.5124 

April 16, 420 hours after the start 0.5272 

April 21, 528 hours after the start 0.530 

Prom this moment (twenty-second day) the ammonia aug- 
ments no more. In a similar medium the nitrogen eliminated 
in the state of ammonia is then from 70 per 100 of the total of 
the nitrogen, instead of 91 per 100 in the culture of the aspara- 
gine. On the other hand we observe that the weight of the 



THE BACTERIOLOGICAL WORLD. 467 

microbes is very considerable; it is of. 0.990 per litre (0.420 
in .the asparagine culture); the weight of the organic soluble 
matter fixed attains 0.495 (0.330 in the culture of asparagine). 
We may conclude that in order to study these last principles of 
secretion, so interesting from a physiological point , of view, it 
would be preferable to employ the medium of gelatine. In 
this case the formation of pyocyanine is almost wanting. 

Notwithstanding the absence of this pigment, the special 
products, as experiments demonstrate, are active. On the 
indications of Mr. Bouchard we have sought their actions. Not 
being able at the moment to study them separately, we have 
had recourse to the following method followed by this author, 
to measure the toxic properties of the urines. By physical 
means of dissociation, these products have been separated into 
three groups — the volatile, the insoluble in alcohol, and the 
soluble in alcohol, i. e., three extracts. 

A very clear property characterizes the first one, that is, 
the portion separated by distillation. This extract acts on the 
vaso-motors, paralyzes the dilator centers, contracts the vessels 
slightly. 

Introduced in the organism of the rabbit, the matters of the 
second extract, taken again by the water or the glycerine, cause 
diarrhoea, fever, albuminuria, hemorrhages ; they weaken the 
resistance of the vaccinated, are opposed to diapedesis, alter the 
tissues. The heat diminishes their poisonousness. In the lowest 
dose the toxic power is not appreciable, while the vaccinating 
power is noticed. This second extract contains some most 
important secretions ; it is poisonous and produces Vaccination 
by temperature ; we can, nevertheless, diminish its deleterious 
effects in preserving those which are useful. There is not abso- 
lute parallelism between these two properties. 

As to the third extract, it causes convulsions, and, if the 
animal does not succumb immediately, generally the subject 
recovers in the end : while, if. without going to a mortal dose, 
insoluble principles have been used, he becomes sicker and 
sicker. 

This third extract sometimes raises the temperature, but it 
never renders refractory any more than the first. So that there 



468 THE BACTERIOLOGICAL WORLD. 

are some microbic substances which are morbific and not vac- 
cinal. 

We must add that the ordinary principles of the life* of 
bacteria, such as the ammonia, becoming too abundant, may 
enter in line of reckoning of the power of poisoning. 

The diverse propositions advanced are all justified by 
numerous experiments. 



HYGIENIC BACTERIOLOGY. 



CORROSIVE SUBLIMATE AS A DISINFECTANT AGAINST THE 
STAPHYLOCOCCUS PYOGENES AUREUS. 



BY B. MEADE BOLTON", M. D. 

A. C. Abbott ( Johns Hopkins Hospital Bulletin, No, 12, 
April, 1891 ) publishes the results of the test made upon cultures 
of the staphylococcus pyogenes aureus with a 1-1000 solution of 
corrosive sublimate. Abbott finds that the disinfectant power 
of corrosive sublimate in the above concentration, when tested 
by methods which exclude the carrying over of minute quanti- 
ties of the disinfectant, is not so great as has been claimed. 
He holds that, in many of the experiments heretofore made to 
test corrosive sublimate, the latter has been assigned a higher 
rank than it deserves, because some of it has been transferred 
to the culture medium, and has inhibited, but not destroyed, 
the growth. 

His experiments were made upon liquid cultures containing 
sterilized sand. With cultures of this sort he was able, by 
filtration, to get a better distribution of the organisms in the 
liquid, avoiding macroscopic clumps, which might interfere 
with the action of the disinfectant upon the organisms in the 
center. Suspensions in water were also used — also filtered. 
Fresh cultures and fresh solutions of corrosive sublimate were 
used, of course, in every case. 



THE BACTERIOLOGICAL WORLD. 469 

Abbott finds that the number of organisms makes a differ- 
ence in the efficacy of the disinfectant. The greater the 
number of organisms the more difficult the disinfection. Cul- 
tures vary in their resisting power — organismsfrom one culture 
resisting better than those from another. Cultures in beef-tea 
resist better than suspensions in water. Organisms which remain 
alive after the action of the disinfectant are retarded in their 
growth and are weakened in virulence. Corrosive sublimate in 
the proportion of 1-400,000, retarded growth in cultures of 
bouillon containing peptone, 1-600,000 without peptone. The 
staphylococci, which have been attenuated by the action of the 
sublimate, regain their virulence when cultivated for some time 
on ordinary culture media. — Brooklyn MedicalJournal. 



INFLUENZA BACTERIA. 

A recent number of the Pr ogres Medical contains an 
abstract of an article by Professor Babes, which appeared in 
the Centralolatt fur BaJcteriologie und Parasitenkunde on the 
subject of bacteria in influenza. The author has succeeded 
in isolating two varieties of micro-organisms to which he thinks 
that in all probability the pathological element in the causation 
of influenza may be attributed. Those of the first variety 
form short chains, are immobile, and are not stained by 
Gram/s method. Their colonies are small in dimensions, trans- 
parent, and developed through the entire culture media. They 
have been proved capable of producing active pulmonary inflam- 
mations. The bacteria of the second class produce on agar- 
agar colonies that present a dark center and a whitish periph- 
ery. They are also immobile, but, unlike the first variety, are 
colored by Gram's method. In mice, the inoculation of this 
culture produced fatal pneumonia, the bacilli being found in the 
blood-vessels ; and in rabbits a local inflammation resulted. — 
New York Medical Journal. 



470 THE BACTERIOLOGICAL WORLD. 

THE FORM OF DYSENTERY PRODUCED BY THE AMCEBA 

COLI. 



BY Wi T. COUNCILMAN, M. D. 

Dysentery, as a word, was first used in a clinical sense. It 
meant tenesmus. It is an infla mmation of the large intestine. 
The pathological lesions are exceedingly numerous and varied.- 
It would be absurd to suppose that all forms of dysentery come 
from the same cause, just as in pneumonia. The pulse is 
rapid. The case may last several months. In the anatomical 
lesions which are produced there is much variation. Some die 
before ulceration begins, and other cases are not so virulent. 
The lesions are in the submucous tissue. Clinically the disease 
is chronic. It begins with a diarrhoea. It is intermittent in 
character, may last for a month, stop, and then break out 
again. There is no pain m the earlier stages,, but later there is 
pain ; the diarrhoea becomes worse, and there is tenesmus. 
The stools are mixed with blood. There is usually no fever in 
the entire course of the disease, emaciation still goes on. The 
duration may be from three to six months. Towards the last 
the complexion assumes a dull, earthy-like color. There is 
nothing distinctive in the stools; they are always fluid, and 
considerable in amount. When these patients die, the 
anatomical lesions are exceedingly characteristic. There is 
entire absence of diphtheritic exudation, which one finds in the 
acute cases, and the surface of the bowel shows numerous 
ulcerations. These ulcers are relatively small, and the surface 
is mammilated. There is a sm all loss of substance at the apex 
of these elevations, and an abscess cavity ; -and on this a 
depression is filled with an opaque, greyish, gelatinous-looking 
mass. The entire intestine is much thickened, the muscular 
coat thickened Then large ulcers can be found which run 
into each other, so that we may have long, sinuous passages 
running through the intestines in various directions, and the 
membrance becomes undermined, sloughs and falls off. It 
extends to the mucosa and t he muscular coat, and large sloughs 
are cast off. The microscopic characteristic appearance is 
evident. The amount of sloughing is often extreme, some as 
large as the hand. 



THE BACTERIOLOGICAL WORLD. 471 

This is interesting from the complications. The most fre- 
quent is the abscess of the liver. There has been seen at the Johns 
Hopkins Hospital eleven cases of this form, of which five died 
and three are still under treatment. Of these eleven cases, 
four deaths occurred from liver abscess, and one case of liver 
abscess is still under treatment. The abscesses of the liver are 
fully as characteristic as in the intestinal ulceration. The 
liver abscesses are filled with a gelatinous-looking mass. If 
they are larger they may have a distinct fibrous form, and look 
like other forms of abscess. 

The next most common complication is abscess of the 
lung. The formation of abscess of the lung is interesting. ■ In 
four cases at the hospital, three had abscesses of the lung. 
The lung abscess comes from the liver abscess. The liver 
abscess is on the upper surface of the liver. There is an 
adhesive pleuritis and the lung abscess follows. It is very 
insidious in its outset, there is always pyrexia, abdominal pain 
and much sweating. When the lung complication takes place, 
the appearance of the sputum is characteristic — tough, of 
dingy cherry-red color, as in acute pneumonia, so that the 
spit-cup can be turned up without the sputum running out. 
The sputum also contains small masses of necrotic tissue, lung 
tissue. The cause of the disease has long been suspected, 
Davaine, in 1853, described the cercomonas in the stools. Then 
Lander, in Prague, found small amoeba? in the stools of a child 
which had died of dysentery, also in cholera. Cholera patients 
in India by Cunningham and Lewis. Lsrch, of St. Petersburg, 
was the first to describe this as we now know it. It is an 
amoeba l0 1 , to ^^ of an inch in length. It is either round 
or is undergoing movements. The outer part is a homogeneous 
mass, and the inner part is filled with granular matter. The 
movements are active, and its large size makes it easy to study 
under a low-power microscope. It may be seen to change its 
form and shape under the microscope. It puts out its 
pseudopodia and draws them in again. It is found in the 
stools, in the abscess of the liver and lung. We find in it 
things which the organism has taken up for food, as for 
example, red blood corpuscles. The organism may be seen in 
the microphotographs, which aue passed around. He thought 



472 THE BACTERIOLOGICAL WORLD. 

this form of disease was not so uncommon as many thought. 
He had tried to find the geographical range of the disease. 
Woodward also had written largely on dysentery, had recorded 
six hundred and ninety-three deaths from intestinal affection. 
— Journal of the American Medical Association. 



MEDICAL BACTERIOLOGY. 



THE TREATMENT OF MALARIA. 

Laveran finds that in the contest between the organism 
and the hematozoons, the leucocytes play an important role. 
It is a well-recognized fact that, if an insoluble and finely- 
pulverized substance be injected into the blood of an animal, 
the leucocytes rapidly invade this powder and charge them- 
selves with its elimination from the blood ( Ponfick, Hoffman, 
Langerhaus) . 

The researches of Metschnikoff prove that the leucocytes 
not only invade inert powders and the bodies of microbes, but 
also, in a great number of cases, the living pathogenic microbes, 
and that this property, known as phagocytosis, plays a very 
important role in the evolution of infectious diseases and in 
the phenomenon of immunity. The cells of the splenic pulp 
and those of the endothelium of the connective tissue, and of 
the pulmonary epithelium participate in this power. He has 
also concluded that the paroxysm of the fever persists during 
the life of these microbes in the blood, ceasing only when the 
parasites become the prey of the phagocytes. Especially is this 
marked in relapsing and malarial fever. The resistance which 
the microbes exert against this action of the leucocytes may be 
of long duration if they be lodged in the spleen or in the 
marrow of bone. 

Wyssokowitsch has noticed that the spores of the bacillus 
subtilis lodged in the spleen were still living at the end of three 
months, an interesting fact in a point of view of the study of 
latent microbism. 



THE BACTERIOLOGICAL WORLD. 473 

Metschnikoff, G-olgi Gamaleia and others insist upon the 
destructive action of heat' upon the microbes, while, on the 
contrary, cold diminishes the activity of the phagocytes. It is 
well known that an organism placed. in the best general condi- 
tion will resist most successfully the inroads of the pathogenic 
microbes, whilst all debilitating causes militate against the cure 
of malaria. Laveran himself has observed in animals whose 
blood contained haematozoa, that under the influence of an 
abundant alimentation the microbes disappeared, while dilata- 
tion favored their development. The anaemic diseases — over- 
fatigue, ill nourishment — are attended with frequent relapses 
of the fever, and the salts of quinine themselves seem to 
become powerless in preventing these relapses. On the con- 
trary, under the influence of only a good regimen and rest, 
frequently the condition of malarial patients will ameliorate, 
the anaemia grow less, the relapses less frequent, and a cure may 
occur without the intervention of a more active medication, 
simply because the organism has been placed in the best hygi- 
enic condition. 

All the tonics have, consequently, a favorable action upon 
the progress of the disease, and foremost among these may be 
placed wine, coffee and arsenious acid. Arsenic taken in 
small doses is of great service in the treatment of rebellious 
fevers with the malarial cachexia. If given in large doses, 
however, relapses of fever will often be produced, and the 
condition of the patient aggravated. Hydrotherapy is valuable, 
always used as a tonic medication. It succeeds sometimes when 
other medications have failed. It is well to know, however, 
that the first treatment may provoke a relapse ; hence it is 
necessary to proceed with prudence, beginning with short and 
lukewarm douches before arriving at cold douches, not touching 
at first the spleen, and prescribing some doses of quinine at the 
same time. One can readily understand that a cold douche, 
especially if applied over the splenic region or following a par- 
oxysm, will cause contraction of the splenic vessels, driving 
the haematozoa into the circulation, The malarial patients, 
who in thermal stations — Vichy, for instance — take these 
waters both internally and in the form of baths, often have 
relapses at the beginning of the treatment, when they do not 



474 THE BACTERIOLOGICAL WORLD. 

proceed with prudence. The change of climate is often bene- 
ficial to these patients, because the great heat of malarial 
climates is very debilitating, giving rise to exhausting sweats, 
nights without rest, and dyspepsia — all of which are corrected 
in a better climate. Quinine is of invaluable service in the 
treatment of these fevers, especially acting as a specific in the 
chronic forms. It has a very powerful destructive action upon 
all microbes, especially upon the protozoa, to which group the 
haematozoa belong. These rapidly disappear from the blood of 
patients who are submitted to a course of quinine medication. 
The spherical bodies and the flagellate forms disappear first. 
The treatment must be continued for some time to insure entire 
destruction of the microbes. It is probable that the phagocytes 
invade more readily the hasmatozoa when they have been killed 
or benumbed by the salts or quinine, but that the activity of 
the phagocytes is not directly influenced by the quinine. — La 
Medicine Moderne, February 19, 1891. 



THE THERAPY OF DIPHTHERIA. 



BY F. LOEFFLER, M. D. 

In our fight against this microbe in nature, two points are 
always before us ; First. To guard against the colonization 
of the bacillus ; in healthy subjects on the intact mucous mem- 
branes, and in diseased individuals on the mucous membrane 
that has not been as yet infected. This colonization can be 
hindered, if we bring the healthy mucous membrane into con- 
tact with some substance which will prevent the growth of the 
bacilli, or, better still, which will kill them, as soon as it is 
brought into contact with them. Of course, these substances 
cannot be of a nature calculated to injure the mucous mem- 
brane, or such that even a continued use of them could give rise 
to toxic or other unpleasant symptoms. Second. The clusters 
of developed bacilli in the superficial layers of the pseudo- 
membrane must be killed, so that no spreading of the infection 
to parts of the mucous membrane, as yet intact, can take place, 



THE BACTERIOLOGICAL WORLD. 475 

and also that no living bacillus may be expelled from the 
patient's mouth, so that the spread of the disease in other per- 
sons may be avoided. 

In experimental work the bacilli should be placed in a 
nutritive substance congenial to them, and in which they will 
increase rapidly, as is the case when they are in the human 
mucous membrane. The temperature of the culture should 
also be such as to facilitate the growth of the germs. I have 
found the following mixture especially favorable to the growth 
of the bacilli : Four parts of beef -blood serum, one pint of 
neutralized bouillon, 1 per cent, of peptone, 1 per cent, of grape 
sugar, 0.5 per cent, of common salt, at a temperature of 
99.5° F. 

Ten cubic centimetres of this serum mixture is placed in a 
test-tube. The tube is held at an angle so as to allow a larger 
surface of the serum to be exposed to the air until it coagulates, 
This tube was now inoculated superficially with a fresh culture 
of the bacillus. In eight hours small colonies could be seen, 
and in twenty-four hours the whole surface of the serum was 
covered with a thick layer of the bacilli. Into the freshly inoc- 
ulated tube > enough of the remedy to be tested was poured to 
cover the surface of the serum — about 10 to 15 cubic centi- ' 
metres. The fluid was either immediately poured off again, or 
else allowed to stay in contact with the serum for ten seconds. 
The act of gargling can seldom be continued longer than ten 
or thirty seconds, and, therefore, this time was chosen in order 
to prove the efficacy of the drugs to be employed. The pouring 
in and out of the fluid did not work off the bacilli. 

If a blood-serum culture is allowed to remain undisturbed 
for several days, it will be found that a thick layer has formed 
over the serum. This layer corresponds with the colonies of 
bacilli in the superficial membranous layers in the human sub- 
ject. After the remedy had been tested upon the superficial 
culture of the bacilli, it was also tested in cultures several days 
old, in a similar manner. The time allowed for contact being 
twenty to thirty seconds. After the fluid had been poured off, 
tests were made both from the superficial and deep layers of 
the culture. 



476 THE BACTERIOLOGICAL WORLD. 

I discovered, in this way, that, while some remedies cause 
the immediate death of the bacilli in superficial cultures, they 
have comparatively little or no effect when the cultures have 
become thick layers, even when the contact is prolonged, and 
that in order to produce any effect the remedy had to be used 
in a concentrated form. 

These comparative experiments seem, to me, to be of great 
practical value. They teach us that a prophylactic treatment 
with weak solutions is not sufficient, but that we must combine 
these with concentrated applications which will also cause the 
immediate destruction of the bacilli in the deeper layers. 

The remedies which I have tested comprise most of those 
which have found, or still find, therapeutic application in diph- 
theria, and also other antiseptics. Some I have subjected to a 
most careful study, others I have only been able to pay passing 
attention to. 

They are as follows : Bichloride of mercury, cyanide of 
mercury, metallic mercury, nitrate of silver, chloride of silver 
dissolved in sub-sulphurate of soda, permanganate of potash, 
chlorate of potash, saturated lime water, aqueous chlorine solu- 
tions, aqueous bromine solutions, aqueous iodine solutions, 
trichloride of iodine, chlorate of lime solutions, double iodide 
of potash solutions, alcohol, allyl-alcohol, benzyl-alcohol, ether, 
chloroform, carbolic acid, salicylic acid, resorcin, hydrochinon, 
tropseolin, benzol, brenzcatechin, the xyloles, toluol, anisol, 
phenetol, pseudocumol, oil of turpentine, anilin, thymol, the 
creosoles, creosol-sulphonic acids, and thirty-eight etheric oils. 

The majority of the organic preparations were placed at 
my disposal by Dr. Limpricht, of our chemical institute, and 
my thanks are due him. 

Bichloride of mercury proved its powerful antiseptic quali- 
ties also in killing the tubercle bacillus. Solutions of 1-10,000 
caused immediate sterilization of a superficial culture, and even 
a solution of 1-20,000, when brought into contact with the 
cultures, caused the death of nearly all the bacilli, the few 
remaining ones, however, formed themselves into colonies 
within a few days. A 1-2,000 solution, even, when kept in 
contact with the cultures for twenty seconds, failed to exert 
any lasting influence upon the deeper layers of the culture. 



THE BACTERIOLOGICAL WORLD. 477 

The employment of a 1-1,000 solution caused the disappear- 
ance of nearly all the bacilli, and any stronger solution than 
this caused their complete extermination, even in the deepest 
layers. 

My experiments with the cyanide of mercury were particu- 
larly interesting. This drug has been used in the treatment of 
diphtheria by Hugo Schulz, Erichsen, Eothe and others. 
Sellden, of Sweden, has, perhaps, had the most extended 
experience with it. He has used it in one hundred and fifty- 
six cases of diphtheria, and only lost four, three of which he 
came too late to relieve. 

His treatment consisted in giving a spoonful of the follow- 
ing formula every hour, day and night : 

Hydrag cyan gram, 0.02. 

Tinct. aconiti grams, 2.00. 

Mel. crud " 50.00. 

Ag. dest " 150.00. 

M. 
For young children 0.1 to 0.4 milligrams of the cyanide 
mercury was the dose employed. Then, two to four times 
during an hour the following gargle was employed : 

Hydrag cyan grams, 0.04. 

Ag. menth. pip " 400.00. 

Either of these solutions have a strength of 1-10,000. My 
own experiences have shown that solutions of 1-10,000 exter- 
minate nearly all the germs, although the contact may only be 
momentary. In solutions of 1-8,000, the sterilization is abso- 
lutely complete. The thick cultures were not, however, steril- 
ized even by twenty seconds'' contact with a 1-1,000 solution, 
this result being obtainable only when a solution of 1-200 was 
used. All the same, repeated applications of a 1-1,000 solution 
will cause subsequent sterilization. Sellden's method of using 
the solutions every fifteen minutes or half hour would surely 
hinder the growth of the bacilli. This treatment, however, 
will not cause a destruction of the bacilli within the membranes. 
In one case of diphtheria I was able to trace the bacilli in the 
membranes ( by cultures) after the cyanide of mercury solutions 
had been used for three weeks. 



478 THE BACTERIOLOGICAL WORLD. 

In my opinion, preparations of silver promise excellent 
results. Nitrate of silver in solutions of 1-1,000, when brought 
in contact with cultures for one to ten seconds, had a decided 
action. The serum would remain apparently sterile for about 
twenty-four hours, then, however, colonies would begin to form 
again. Stronger solutions of 1-150 caused complete destruction 
of the bacilli. The same solution had no effect upon cultures, 
even when kept in contact for twenty seconds. 

Fresh chloride of silver, dissolved in sub-sulphurate of 
soda, was found by Behring to be very destructive to the gono- 
coccus of gonorrhoea in solutions of 1-7,500. Personally I 
brought four cases of whooping cough to a speedy termination 
by inhalations of a 1-10,000 solution of the chloride. Solu- 
tions stronger than 1-5,000 are very irritating. In experiment- 
ing with this drug on serum infected with the bacillus of 
diphtheria, I found that ten seconds' contact with solutions of 
strengths, varying from 1-600 to 1-4,000, rendered the serum 
sterile, while the tubes tested with solutions of 1-5,000 to 
1-10,000 developed small colonies the subsequent day. But on 
the third or fourth day, all the tests contained bacilli. In the tube 
treated with the 1-600 solution, the colonies were very mark- 
edly disseminated, but in the weaker solution a coating of the 
bacilli had formed over the serum. Indeed, this drug showed 
no superiority to nitrate of silver. 

Permanganate of potash in a 1-per-cent. solution left 
enough bacilli to form new colonies within the course of a day. 
A 2-per-cent. solution sterilized the surface of the serum. In 
order to positively destroy cultures within twenty seconds, a 
5-per-cent. solution was necessary, although the 3-per-cent. 
solution was very active. 

Chlorate of potash, 5 per cent., saturated lime water, 
peroxide of hydrogen 1 per cent., sulphuric acid 1.25 per cent., 
and lactic acid 1 per cent., were entirely inactive when kept in 
contact for 10 seconds. 

Iodine in aqueous solutions was inactive, but a solution 
composed of 5 parts of iodine, 10 parts of iodide of potash, and 
300 parts water, sterilized cultures in 20 seconds. 

Bromine in 1-300 to 1-500 solutions sterilized the serum 
in 10 seconds. The bacilli in cultures were only destroyed 



THE BACTERIOLOGICAL WORLD. 479 

after a 20 seconds' contact with a 215-per-cent. solution of 
bromine. 

Chlorine water, containing 0.009 grams of 01. to the 
cubic centimetre (1-112.7) destroyed the bacilli on the serum, 
even in weaker solutions (1-1,100) in 10 seconds. The first- 
named solution also sterilized cultures in 20 seconds, but the 
weaker solutions had but little effect. 

Although saturated lime water had been proven inactive, 
saturated solutions of chlorate of lime were most effective. 
Fifty parts of chlorate of lime were mixed with 200 parts of 
water. The filtrate destroyed the bacilli on the serum, even 
when diluted twenty-five times. The cultures, however, were 
only destroyed by the undiluted filtrate, in 20 seconds. Weaker 
solutions were almost inactive. 

Behring has recently used trichloride of iodine successfully 
in treating diphtheria in animals, in a 1-1,000 solution. Even 
a 1-2,000 solution has a decided action. My own experiments 
showed that a 1-per-cent. solution exterminates the bacilli in 
cultures in 20 seconds. 

Absolute alcohol destroyed nearly all the germs on the 
serum surface upon momentary contact, and even a 5-per~cent. 
solution was decidedly active. A mixture of alcohol and ether 
is especially valuable, and the following mixture I have proved 
to be most serviceable : 

ft 

Alcohol 50 parts. 

Ether 25 parts. 

Water 25 parts. 

On cultures, however, both alcohol and ether gave negative 
results. 

Allyl-alcohol gave negative results, while benzyl-alcohol 
sterilized the inoculated serum in 10 seconds completely. 
Chloroform water was equally as effectual. 

Carbolic acid gave negative results in a 1-per-cent. solution* 
A 3-per-cent. or 4-per-cent. solution will sterilize the inoculated 
serum. The addition of 20-40 per cent, of alcohol greatly 
intensified the action of a 2-per-cent. solution. Cultures were 
destroyed in 20 seconds by a 5-per-cent. solution. The same 
results could be accomplished by a 3-per-cent. solution, with 
the addition of 30 per cent, of alcohol. 



480 THE BACTERIOLOGICAL WORLD. 

Lysol, which has been recently introduced into commerce, 
proved very efficacious. Two-per-cent. solutions sterilized the 
inoculated serum in 10 seconds. Cultures, however, were not 
positively sterilized, even by 20 seconds' contact with a 5 per- 
cent, solution. 

The kresols were but slightly active. Salicylic acid, while 
active in sterilizing serum inoculations, gave negative results in 
cultures. Two and five-per-cent. solutions were employed. 

The action of the etherial oils was especially interesting. 
The majority of these strongly odorous oils contain carbo- 
hydrogen in the form of C 10 H 16 , and a few contain oxygen. 
These actions were extremely varied. The majority of them, 
however, seemed to check the growth of the bacilli, even by 
their fumes. Anisol in a 5-per-cent. solution of equal parts 
of ether and alcohol was very effective. Thymol in steam form 
was inactive, but in a mixture with 20-per-cent. of alcohol, it 
killed all the bacilli upon contact — even in solutions of 1-500. 
The direct application of watery solutions of anilin were active 
on the serum surfaces. Anisol solutions were inactive. Oil of 
turpentine, even with the addition of 50 per cent, of alcohol, 
gave negative results, but the addition of 2 per cent, of car- 
bolic acid rendered it very active. 

Metallic mercury in vapor form was effective in sterilizing 
serum surfaces. 

So much for my experiments, which, however, are far from 
being complete. Many of the most active agencies are, unfor- 
tunately, unsuitable for practical use, on account of their toxic 
qualities. 

For prophylactic treatment of persons exposed to diph- 
theria, gargling every two or three hours, for five or ten 
seconds with a suitable solution, is recommended. For this 
purpose, I have found the following very useful : Bichloride 
of mercury 1-10,000, or 1-15,000, or better still, cyanide of 
mercury in a 1-8,000 solution, or 1-10,000. The taste of the 
latter preparation is less unpleasant than that of the former. 
Chloroform water, or thymol 1-500 in 20 per cent, of alcohol, 
is also useful and agreeable. It is of great importance that 
the gargling should be done properly and thoroughly. In vapor 
form, oil of orange peel, lemon oil, eucalyptus oil, anisol, phen- 



THE BACTERIOLOGICAL WORLD. 481 

etol, benzol and toluol, are of service. These are best used 
according to Feldbausch's method. A small metallic tube, 
containing absorbent cotton, moistened with the oil, is inserted 
in each nostril, and the patient is therefore continually breath- 
ing the fumes. These tubes should remain in place as long as 
the patient is in an atmosphere probably victated with the 
bacilli of diphtheria. The same oils might also be sprayed or 
vaporized in the sick room with material advantage. In this 
way the air would be more or less saturated with the fumes, 
and, certainly, an atmosphere in which orange peel oil, or 
lemon oil, is suspended, is both pleasant and refreshing. 

Eegarding the treatment of patients actually suffering 
from this dread disease, I advocate gargling every one or two 
hours with weak solutions, and gargling every three or four 
hours with solutions which will positively destroy the bacilli, 
even in thick cultures, For the latter purpose, I have found 
the following most practicable : Bichloride of mercury, 
1-1,000 ; carbolic acid 3-per-cent. solution in 30 per cent, of 
alcohol, finally, equal parts of alcohol and oil of turpentine, 
with 2 per cent, of carbolic acid. Between the appointed hours 
for gargling, the parts might be painted with a 5-per-cent. 
solution of carbolic acid, 2 per cent, of bromine, 1 per cent, of 
chlorine, or with a concentrated watery solution of the kresols, 
especially o-kresol and^?-kresol. 

Bichloride of mercury has been used with the most excel- 
lent effects, and also the 3-per-cent. solution of carbolic acid in 
30 per cent, of alcohol. Under the treatment described above, 
the membranes can be proven to be sterile within a few days. 
The general condition of the patient improves rapidly. Garg- 
ling with the carbolic-acid solution will at first be unpleasant 
to the patient, but this soon passes off, and the patient soon 
becomes accustomed to it. It is to be noted that, in diph- 
theritic scarlet fever, the above treatment is also very 
successful. 

I trust, that, in this brief lecture, I have been able to set 
forth clearly the principles of the modern therapy of diphtheria, 
based on the most recent researches and extensive experience, 
and that with the advance of medical knowledge this bete noxr 
of medicine may lose many of its horrors, and be able to claim 
but few victims. — Exchange. 



482 THE BACTERIOLOGICAL WORLD. 



SELECTIONS. 



STUDIES ON THE ACTION OF DEAD BACTERIA IN THE 

LIVING BODY. 



BY T. MITCHELL PRUDDEN, M. D. 

Director of the Laboratory of the Alumni Association, College of 
Physicians and Surgeons, New York. 

AND EUGENE HODENPYL, M. D. 

First Assistant in Pathology. 

SEC.OND ARTICLE. — THE ACTIOK OF STERILIZED CULTURES OF 
THE TUBERCLE BACILLUS. 

. We have seen in the first article that the action of bacteria 
in the body is by no means limited to the changes induced by 
those substances called ptomaines, albumoses, or toxalbumins, 
which are set free by living germs. It appears that the proteid 
constituents of the bacterial cells themselves, when these are 
set free, either by a natural disintegration of the germs or by 
an artificial extraction, are capable not only of stimulating the 
fixed body cells to proliferative changes, but may, by calling 
into play the forces involved in chemotaxis, cause, either a 
moderate collection of leucocytes or induce marked suppu- 
ration. 

It is probable that these two fairly distinct factors, the 
eliminated ptomaines and albumoses on the one hand, and the 
bacterio-proteins on the other, are both involved in many, if 
not most, of the acute infectious diseases. But one or other 
of these factors is apt to be so preponderant in many cases as 
to fairly dominate both the symptoms and lesions. 

For example in tetanus, typhoid fever and Asiatic cholera, 
the toxic substances absorbed into the body at large from the 
seat of growth of the pathogenic germs may determine pro- 
found and even fatal symptoms without much local change at 
the proliferating germ centers. 



THE BACTEKIOLOGICAL WORLD. 483 

On the other hand, in many of the local suppurations — in 
pneumonia and in tuberculosis, for example — the most marked 
effects are apt to be induced in the immediate vicinity of the 
invading germs. 

In still other diseases — diphtheria, for example — we may 
have extreme local tissue changes together with profound sys- 
temic effects. 

Finally, under a variety of unusual conditions, pathogenic 
germs which commonly induce one particular order of lesions 
may bring about most striking alterations of another class. 

As an example of this variability of effect, we may cite the 
suppurative inflammations which not infrequently occur as com- 
plications of typhoid fever, when these germs lodge or grow in 
unusual situations. 

Perhaps, the most striking example of a pathogenic germ 
whose effects are closely limited to the immediate neighbor- 
hood of its seat of growth is the bacillus tuberculosis. That 
there may be systemic effects induced by the absorption into 
the body at large of poisons eliminated by the tubercle bacillus, 
we, of course, cannot deny. But, if such there be, we certainly 
know very little about them, and they must be insignificant in 
comparison with the dominant lesion of tuberculosis — namely, 
the production immediately about the germs of a new short- 
lived tissue having a moderately characteristic morphology and 
prone to undergo degenerative changes of great significance, 
both to the germs which cause it and to the integrity of the 
invaded organ. 

It is because of the peculiarly direct and constant relation- 
ship between the germ and the lesions which it induces that we 
have selected the tubercle bacillus for an experimental study 
along the lines which our preliminary paper has suggested. 

But, before detailing our own experiments, it seems desir- 
able to briefly notice some observations already made in this 
field, which will have a bearing in establishing our point of 
view. 

A number of studies have been made on the metabolic pro- 
ducts set free during the growth of the tubercle bacillus, both 
in cultures and in the body. But these studies have been 
made from such a variety of different standpoints and with 



4.8-i THE BACTERIOLOGICAL WORLD. 

such different technique that it is difficult to glean any very- 
positive and definite data from the results. 

Hammerschlag found in the alcohol and ether extract of 
the tubercle bacilli, fat, lecithin and a poison which induced 
spasms and death in rabbits and guinea-pigs. The residual 
material of the bacilli contained an albumenoid and cellulose. 
He could not separate a ptomaine by Brieger's method, but did 
find evidence of a fever- causing toxalbumiu. 

The recent publications of Koch on the nature and effects 
of substances in or derived from cultures of the tubercle bacillus 
we may assume to be familiar to all our readers. But the 
manner in which they have been made known and the lack of 
detail in the description of the mode of preparation of his 
" lymph " leave his results in a most unsatisfactory state of 
indefiniteness, so far as the scientific bearing of the subject is 
concerned. Some substance or substances, it would appear, 
either set free during the growth of the tubercle bacilli or, as it 
is said, extracted from them by glycerine, are capable of exert- 
ing rather ill-understood effects upon tubercle tissue in the 
body and are not without extreme effects in many cases upon 
the body at large. With the therapeutic bearing of the sub- 
stances contained in Koch's tuberculin we have here nothing 
to do. 

One point, however, in Koch's last publication on this 
subject has an important bearing upon the subject in hand — 
namely, the statement that pure cultures of the tubercle 
bacillus killed by boiling, or in other ways, are capable of 
inducing local suppuration with no other effects, if introduced 
subcutaneously into the guinea-pig. 

Hueppe and Scholl cultivated the tubercle bacilli in a 
glycerine-peptone bouillon, and found that the fluid contained 
substances having essentially the same effects upon animals as 
those noted by Koch. As the result of an interesting series of 
experiments, these observers came to the conclusion that the 
specific poison of Koch's "lymph" did not belong to the pro- 
teins, but to some substance or substances eliminated by the 
bacilli during their growth and contained in the culture fluids, 
and which were not, as Koch asserted, extracted from the 
culture by glycerine. Hueppe and Scholl separated from the 



THE BACTERIOLOGICAL WORLD. 485 

" lymph " which they had prepared, and which seemed to be 
practically identical with the " lymph " of Koch, by precipita- 
tion with alcohol, a substance which appears to contain the 
active ingredient of the " lymph, " They separated in the 
same way the same substances from Koch's " lymph," and, 
on testing these in tubes beneath the skin of animals, they 
found that in both cases this substance was markedly chemo- 
tactic. 

That Koch's lymph is not, as it is furnished in the crude 
condition, chemotactic and does not tend to cause abscesses, 
may be due to the fact that it contains glycerine, which exerts 
an influence on the leucocytes opposite to that which positive 
chemotactic substances display. 

Trudeau, wording also with fluid cultures, obtained 
materials apparently the lesult of metabolic processes of the 
tubercle bacilli which induced toxic effects upon animals. But 
these were not marked, since he did not succeed in sufficiently 
concentrating the fluid containing the materials sought for. 

Zulzer extracted from agar cultures a substance which, on 
purification and injection subcutaneously in small quantities 
into rabbits and guinea-pigs, caused temporary increased 
rapidity of respiration and an elevation of temperature. Larger 
doses were fatal. 

Crookshank separated the bacilli from glycerine-broth 
cultures by filtration, and concentrated the filtrate by evapora- 
tion. This concentrated filtrate, injected subcutaneously into 
guinea-pigs, induced muscular spasms, a lowering of the tem- 
perature, and death. 

Weyl extracted tubercle bacilli with dilute caustic soda, 
and found in this extract a substance which seemed to him to 
belong in the mucin group. Subcutaneous injection of a solu- 
tion of this substance produced in animals a local necrosis at 
the seat of injection. 

Maffucci found that sterilized cultures of tubercle bacilli 
introduced beneath the skin of guinea-pigs in considerable 
amounts were capable of inducing a chronic poisoning of the 
organism, ending in death from marasmus and great destruc- 
tion of the red blood cells in the spleen, but does not note any 
marked local changes beyond the producing of an abscess. 



486 THE BACTERIOLOGICAL WORLD. 

Wyssokowicz killed the tubercle bacilli in cultures by boil- 
ing, and these on subcutaneous injection in a rat caused a small 
local abscess, containing well-formed and readily stained though 
dead tubercle bacilli. 

An emulsion of a boiled culture of the tubercle bacilli 
was injected into the abdominal cavity of a rat. The animal 
was killed on the thirtieth day. A few small nodules were 
found beneath the liver which consisted of a central pus-like 
mass surrounded by a thin new-formed tissue wall. The central 
portion of these nodules consisted of tubercle bacilli and 
leucocytes, while their walls were composed largely of endothe- 
lial cells, with here and there giant cells among them. Readily 
stained tubercle bacilli were scattered through the walls of these 
nodules, and the epithelioid and giant cells were often grouped 
about them. The microscopical examination of the liver 
revealed numerous small spheroidal nodules composed of 
epithelioid cells and leucocytes, among which were scattered a 
few tubercle bacilli. 

We know that various kinds of substances, organic and 
inorganic, when introduced into the body, can become sur- 
rounded by a tissue in many respects resembling tubercle tissue. 
This was abundantly shown in the years just preceding the 
discovery of the tubercle bacillus and the final establishment 
of tuberculosis as an acute infectious disease. Whether the 
dead tubercle bacilli in these experiments of Wyssokowicz's act 
in some special way in stimulating the tissues to these tubercle- 
like growths, or whether they do this simply as foreign bodies, 
must be shown by further studies. 

The action of the living tubercle bacillus in inducing the 
local lesions of tuberculosis seems to be quite complex. But 
there appear to be several tolerably distinct classes of influences 
at work. 

In the first place, there seems to be an irritation or stimu- 
lation of the tissues near the bacilli which results in the prolif- 
eration of cells, primarily and notably of the connective tissue 
cells. The effect of this is to produce what is called the 
epithelioid cell tissue. Following this, or associated with it, 
there is a local action either on the leucocytes or the blood 
vessels, or both, which leads to the accumulation of small 



THE BACTERIOLOGICAL WORLD. 487 

spheroidal cells either in the periphery of the tubercle or dis- 
tributed through it. Generally there is a necrotizing action 
which results in the coagulation necrosis or cheesy degeneration 
of the new-formed tissue or other tissue near the tubercle. 

The giant cells which form such striking features in many 
phases of tuberculosis appear to be formed under the influence, 
on the one hand, of those factors which induce cell proliferation, 
and, on the other, of those which retard the completion of the 
cell proliferation when once it is under way. The impulse to 
cell division under these circumstances is in so far successful 
that a protoplasmic mass is formed with many nuclei, bat the 
consummation of this attempt, the division of the cell bodies, 
is in abeyance. 

In view of the new facts regarding the singular power of 
the bacterio-protein and chemotaxis which the researches briefly 
summarized in our first article have revealed, and the promising 
but, it must be confessed, rather obscure light which the recent 
studies on the poisons of the tubercle bacillus have elicited, it 
has seemed to us that some further studies on the tubercle 
bacillus should be made with the view of learning, if possible, 
in more detail something of the factors which are concerned in 
developing the complex structures which we call tubercles and 
tubercle tissue. 

Our first series of studies was made with the view of 
determining what effects, if any, are produced in the body of 
the rabbit by the introduction of cultures of the tubercle 
bacilli killed* by prolonged boiling and freed from any material 
elaborated by them during their growth on the artificial culture 
media, so far as such substances are soluble in the culture 
media, or in water, or in 50-per-cent. glycerine. That is to 
say, we wished in these experiments to learn whether dead 
bodies of the tubercle bacilli alone, apart from any of their 
elimination or soluble metabolic products, produced changes 
in the body cells of living rabbits, and, if so, of what kind. 

Technique. — We cultivated the tubercle bacillus for this 
purpose on peptone agar with 6-per-cent. glycerine, and in 
glycerine- peptone bouillon. In all cases we continued the 
growth until it became voluminous, and we obtained identical 
results, whether the cultures were two months or six months 



488 THE BACTERIOLOGICAL WORLD. 

old, and whether they were made in bouillon or on agar. We 
have used cultures of tubercle bacilli derived from three differ- 
ent sources. 

The solid masses of culture were scraped carefully off 
from the agar, or filtered from the culture bouillon. In some 
cases we were not particular to remove from the cultures what 
culture bouillon clung to the germ masses. But in most of the 
experiments the masses of bacilli were washed with sterilized 
distilled water, and then boiled in a small amount of sterilized 
water for from an hour and a half to four hours. In still 
another set of experiments we first carefully washed the culture 
masses, and then boiled them for from two to four hours in 
50-per-cent, glycerine with water, and then filtered off the fluid, 
washing the germ masses before making an emulsion with pure 
water for injection. 

We have found that the condensation fluid in glycerine- 
agar cultures and also the glycerine bouillon in *which the 
tubercle bacilli had grown, when dropped into strong alcohol, 
gives a white precipitate, which, according to the studies of 
Koch and Hueppe and Scholl, contains the active principles of 
Koch's tuberculin. We furthermore found that, after the 
glycerine-agar condensation water or the glycerine bouillon in 
which the tubercle bacilli are grown has been carefully washed 
out of the culture masses of bacilli, no more of this tuberculin- 
containing material is extracted from the bacilli by either water 
or 50-per-cent. glycerine, even after boiling for four hours. It 
would thus appear that the active substance in Koch's ' ' lymph " 
is a material elaborated by the life processes of these germs and 
set free in the culture media, as shown by Hueppe and 
Scholl ( 3 ). 

After steaming the bacilli for from an hour and a half to 
four hours in the steam sterilizer, and separating them from 
their metabolic products in the manner described, we have 
made an emulsion of the bacilli in sterilized water which we 
have used in our experiments. In a few cases we have boiled 
the bacilli together with a small amount of the culture fluids 
containing their metabolic products, but the results of these 
experiments have been the same as when we have carefully 
separated them. 



THE BACTERIOLOGICAL WORLD, 489 

"We find that after prolonged boiling, either in water or in 
50-per-cent. glycerine, while many of the bacilli present the 
usual bizarre, broken, granular involution and degeneration 
forms so often described, most of them preserve their form 
intact, and are stained in the usual way by the Koch-Ehrlich 
method. The broken granular forms are readily stained. 

We have, by a series of separate experiments, proved that 
the tubercle bacilli are certainly killed by the prolonged steaming 
to which they have been subjected, of from an hour and a half 
to four hours. 

As this is, of course, a point of vital importance in our 
experiments, we have been very careful about this preliminary 
sterilizing. We have used the Arnold steam sterilizer, which 
is one of the most efficient forms of laboratory sterilizers known 
to us. The material to be killed has been invariably placed in 
small, thin-walled flasks, exposed to the full action of Jive 
steam. The duration of the steaming was, of course, carefully 
reckoned from the time when the chamber of the steamer and 
its contents was at the full boiling temperature. While we 
have found in a series of preliminary experiments that an hour 
and a half 's steaming sufficed to kill the cultures, as, from 
what we know of the vulnerability of these germs, it should, 
in all except one set of experiments, we continued the steaming 
from two to four hours, the average time being about two hours 
and a half. 

The masses of tubercle bacilli thus steamed have been 
proved to be dead, first, by their failure to grow on favorable 
culture media ; and, second, by, the much more delicate test of 
repeated inoculations in guinea-pigs, which invariably gave 
negative results so far as the development of tuberculosis is 
concerned. Finally, we have inoculated guinea-pigs with 
materials from the lesions about to be described as induced in 
rabbits by the dead cultures, and obtained in the guinea-pig 
only negative results. 

In these ways we have satisfied ourselves that the material 
which we were using contained in every case no living tubercle 
bacilli, but only their dead bodies. 

Experiments. — subcutaneous injections. — We first 
made a series of injections of the emulsion of the sterilized 



490 THE BACTERIOLOGICAL WORLD. 

tubercle bacilli in rabbits beneath the skin and found that in a 
considerable proportion of cases a small local abscess is 
developed in from two to six weeks. This abscess, sometimes 
firmly encapsulated with fibrous tissue, contains pus cells, granu- 
lar detritus, and many readily stained and well-formed tubercle 
bacilli, together with involution forms. This pus, as above 
indicated, on inoculation into that most susceptible animal, the 
guinea-pig, gave negative results ; no tuberculosis developed, 
which was to be expected if the cultures of the tubercle bacilli 
used for inoculation had been readily sterilized. We have 
further made cultures of this pus with a view of determining 
whether this suppuration might not be due to some of the 
ordinary pyogenic bacteria, but the results were negative. We 
thus find, in consonance with the results of Koch, and of 
Hueppe and Scholl, that dead tubercle bacilli are, under certain 
conditions, markedly pyogenic. 

We have tested the chemotactic powers of sterilized cultures 
of the tubercle bacillus introduced in tubes beneath the skin 
of rabbits, in the manner described in our first article, and 
found in every case after six days a plug of leucocytes extend- 
ing deep into the broken end of the tube. We have introduced 
the dead cultures beneath the skin of rabbits inclosed between 
the laminae of Ziegler's plates. In from three to five days the 
capillary space was found crammed with leucocytes. The dead 
tubercle bacillus thus possesses marked positive chemotactic 
powers. 

Peritoneal and pleura injections. — We next made 
injections of the emulsion of dead tubercle bacilli in consid- 
erable quantities (2 to 3 cc. of a milky emulsion) into the 
peritoneal and pleural cavities — in four animals into the peri- 
toneal cavity ; in two, into the pleural cavity. We obtained 
one positive result in each set of injections. The others were 
all negative. 

In the positive cases, whose results were similar in both the 
pleura and peritoneum, we found several larger and smaller 
white nodules adherent to the serous surfaces. These consisted 
of a central soft creamy mass, made up largely of pus cells with 
cell fragments and granular detritus intermingled with large 
and smaller clusters of readily stained and well-formed tubercle 



THE BACTERIOLOGICAL WORLD. 491 

bacilli. While the nuclei of some of the pus cells were not 
readily stained by hematoxylin, there seemed to be no evidence 
of well-marked coagulation necrosis in the cells except when 
very large masses of the dead bacilli were introduced. This 
central pus mass was in all cases closely surrounded and inclosed 
by a wall of fibrous tissue, in the inner layer of which were 
numerous larger and smaller masses of epithelial cells inter- 
mingled with giant cells. Some of these epithelial and giant 
cell islets were irregular in shape and diffuse in outline. Oth- 
ers were sharply circumscribed and rounded, and presented the 
general appearance of miliary tubercles. Tubercle bacilli were 
everywhere abundant in these epithelial cell masses, and 
nowhere else in the fibrous wall of the nodules. Well marked 
cheesy degeneration was nowhere seen in these cell masses 
which resemble tubercle tissue, even when the animal remained 
alive for four weeks after the injection. 

We thus see that the injection of dead cultures of the 
tubercle bacilli is capable of occasionally inducing in the pleural 
and peritoneal cavity of the rabbit a suppurative focus and the 
formation about it of a tissue morphologically similar to tuber- 
cle tissue. 

While these results are interesting, we should not forget 
that a great variety of substances, organized and unorganized, 
on introduction into the serous cavities, as well as beneath the 
skin, are capable of stimulating the cells of the part to the pro" 
duction of a new tissue which in many respects closely resem- 
bles in its morphological characters the tissue induced by living 
tubercle bacilli. The new tissue growths formed under these 
conditions have been the subject of the most extended research 
in times gone by. And every pathologist who lived through 
the epoch immediately preceding the discovery of the tubercle 
bacillus knows to what fallacious conclusions experimental 
studies on the introduction of foreign bodies into animals led 
many an observer, owing to the unwarrantable stress which was 
laid upon epithelioid cell tissue and giant cells in determining 
the morphology of tubercle. 

We have, therefore, left this line of experiment here and 
turned our attention to a form of experiment which seemed to 
promise a more direct insight into the detailed effects of dead 



492 THE BACTERIOLOGICAL WORLD. 

tubercle bacilli upon the living animal cells. This was the injec- 
tion of the dead germs directly into the blood vessels of the 
rabbit through the ear veins. 

The leading idea in these experiments was to introduce into 
the body the dead bacilli under such conditions that their effects 
upon individual cells could, if possible, be determined. 

Intravenous injections. — We have used in these experi- 
ments the sterile emulsions of tubercle bacilli described above, 
and have injected them in moderate quantity, aiming to intro- 
duce into each animal an amount of the material which would 
correspond to a spheroidal mass of the solid culture of from 
about one to two millimetres in diameter, diffused in about two 
cubic centimetres of water. In order to avoid gross vascular 
disturbance it is important to break up as much as is practicable 
the flocculent masses of the tubercle bacilli with a platinum 
needle before injecting them. The animals bear this injection 
into the blood perfectly well, and we have in our series of exper- 
iments in this way rarely observed their general health was in 
any way impaired up to the third week, when a small proportion of 
the animals have died emaciated ( five out of twenty- four ). The 
animals were killed at intervals from the first day to the second 
month, as follows : First, second, fifth, eighth, tenth, seven- 
teenth, eighteenth, twenty-first, twenty-second, twenty-fifth, 
twenty-sixth, twenty-seventh, twenty-eighth, thirty-second, 
thirty-fourth, thirty-fifth, forty-eighth, sixtieth day. The 
organs carefully hardened in strong alcohol, the lungs being 
filled through the trachea. Some of the sections were stained 
in the usual way with hematoxylin and eosin, while others 
were stained for tubercle bacilli. We have experimented in this 
way upon twenty-four rabbits. 

We deem it best to give a summarized statement of our 
results, rather thau a detailed account of the individual experi- 
ments, because the effects of the dead bacilli vary considerably, 
depending upon the amount introduced and the size and vigor 
of the animal, and in many other less well-defined ways. 

If the animal is killed within twenty-four hours after the 
injection of a moderate amount of dead tubercle bacilli, the 
organs hardened in alcohol, and sections finally stained for 
tubercle bacilli, it will be found that the bacilli are present in 



THE BACTERIOLOGICAL WORLD. 493 

by far the largest numbers, and in the largest masses in the 
capillaries of the lungs. They are next most abundant in the 
capillaries of the liver, and may be found in small numbers in 
the spleen. 

After the first three or four days we have been unable to 
find bacilli in the spleen, but they appear to continue in approxi- 
mately the same numbers in the lungs and liver as at first. 

We have made no examinations of the other viscera with a 
view of determining the presence or distribution of the bacilli 
in them. 

We need only refer here to the experiments of Wyssokowicz 
( 12 ) and others on the rapid disappearance of micro-organisms 
injected into the blood.* 

Examinations of sections of the organs of animals killed 
during the first and second week after the injection usually 
show that, while the bacilli have largely disappeared from all 
the viscera except the lungs and liver, they may still be found 
scattered singly or in small clusters in the small blood-vessels of 
these organs. Their situation can usually be best made out in 
the capillaries of the liver. Here they may be lying apparently 
free against the wall of the vessel, or they seem often to be sur- 
rounded by a small quantity of a nearly homogeneous or finely 
granular material which may be fibrin or blood plates, or, finally, 
they may lie singly or in small masses inclosed in variously 
shaped cells. 

The earliest, and throughout the experiments the most 
constant, lesion which we have found "as the result of the venous 
injection of dead tubercle bacilli is the development in the 
lungs of -from one or two to innumerable small white nodules, 
some invisible to the naked eye, some as large as two or three 
millimetres in diameter. These small nodules may appear as 
early as the fifth day after the injection, and persist up to the 
end of the second month, In the earliest days they consist of a 
central denser mass made up of epithelioid and giant cells, 



* The use of masses of dead tubercle bacilli in the study of mu