1
-m
BACTERIOLOGY
MEDICINE AND SURGERY.
A PRACTICAL MANUAL
FOR
PHYSICIANS, HEALTH OFFICERS AND STUDENTS,
BY
WM. HALLOCK PARK, M.D.,
ASSOCIATE PROFESSOR OF BACTERIOLOGY AND HYGIENE, UNIVERSITY AND BELLE-
VUE HOSPITAL MEDICAL COLLEGE, AND ASSISTANT DIRECTOR OF THE
RESEARCH BACTERIOLOGICAL LABORATORIES OF THE DE-
PARTMENT OF HEALTH, CITY OF NEW YORK.
ASSISTED BY
A. R. GUERARD, M.D.,
ASSISTANT BACTERIOLOGIST, DEPARTMENT OF HEALTH, CITY OF NEW YORK.
ILLUSTRATED WITH 87 ENGRAVINGS AND 2 COLORED PLATES.
LEA BROTHERS & CO.,
NEW YORK AND PHILADELPHIA.
BIOLOGY
LIBRARY
G
Entered according to the Act of Congress, in the year 1899, by
LEA BROTHERS & CO.,
In the office of the Librarian of Congress. All rights reserved.
DORNAN, PRINTER.
[
PREFACE.
IN the following pages the attempt has been made
to group together those facts in Bacteriology which
will constitute a sufficient text-book for the student and
which are of direct practical value to the physician
and health officer. Laboratory technique is given in
its essentials and to such an extent as is necessary to
make bacteriological methods plain to the physician,
to guide him in making the simple examinations pos-
sible in his office, and to show him under what
conditions he can obtain diagnostic or other help from
bacteriological examinations in laboratories. The phy-
sician can readily understand and apply the essentials
of bacteriology, but the actual carrying out of the
more difficult examinations should be left to the
trained bacteriologist.
Such subjects as the chemical changes produced by
bacteria, infection, immunity, the nature and use of
protective serums, and the diagnostic value of bacte-
riological cultures, are particularly emphasized, since
knowledge of such subjects is of special importance to
91547
iv PREFACE.
the practising physician, in that it enables him- to ob-
tain an intelligent grasp of the nature of the infectious
diseases.
The methods used in the laboratory for the isolation
and identification of the typhoid, tubercle, and diph-
theria bacilli have been given with especial fulness, as
bacteriological examinations of the discharges of per-
sons suspected to have typhoid fever, tuberculosis, or
diphtheria are now generally made for these bacteria
in the laboratories of the health departments of even
the smaller cities, because of the manifest importance
to the public of knowing where such sources of infec-
tion exist.
In preparing this book the best works have been
freely consulted. Of these, those of Fliigge and
Sternberg, on General Bacteriology, and those of
Abbott and Mallory and Wright, on Technique, should
perhaps be especially mentioned.
My sincere thanks are due to Dr. Hermann M.
Biggs, the Director of the Bacteriological Laboratory,
and to my colleagues in it, who have so freely furnished
me with the results of their original investigations. I
wish also to especially acknowledge my indebtedness
to Dr. A. R. Guerard, who has given me invaluable
aid in the preparation of the book. The illustrations,
with the exception of those on malaria and cholera,
for which I am indebted to Drs. Welch and Dunham,
PEE FA CE.
are almost entirely from photographs taken from
cover-glass preparations and cultures by Dr. Edward
R. Learning, Instructor in Photography in the Medical
Department of Columbia University.
NEW YORK, November,
CONTENTS.
INTRODUCTION.
PAGE
Historical Sketch of the Development of Bacteriology . . 17
CHAPTER I.
The General Characteristics of Bacteria Their Morphology
and Structure Vegetative and Spore Forms ... 33
CHAPTER II.
The Chemical Composition of Bacteria The Conditions Suit-
able for their Growth ....... 50
CHAPTER III.
Vital Phenomena of Bacteria Motion, Heat, and Light Pro-
ductionChemical Effects ( Ferments, Ptomai'ns, Toxins) . 58
CHAPTER IV.
The Relation of Bacteria to Disease ..... 85
CHAPTER V.
Immunity . . . . . . . . . . 102
CHAPTER VI.
Theories of Infection, Immunity, and Recovery . . . 118
CHAPTER VII.
Infection 128
viii CONTENTS.
CHAPTEK VIII.
PAGE
The Effect of Light, Electricity, Pressure, Agitation, Drying,
and Association with Other Micro-organisms upon Bac-
teria 134
CHAPTEK IX.
Effect of Temperature upon Bacteria 144
CHAPTER X.
The Destruction of Bacteria by Chemicals .... 151
CHAPTEK XL
Practical Disinfection and Sterilization (House, Person, In-
struments, and Food) Sterilization of Milk for Feeding
Infants 168
CHAPTEK XII.
The Preparation, Staining, and Microscopical Examination
of Bacteria . 197
CHAPTEK XIII.
The Cultivation of Bacteria Sterilization of Media Appa-
ratus . .212
CHAPTEK XIV.
The Use of Animals for Diagnostic and Test Purposes . . 236
CHAPTER XV.
The Procuring of Material for Bacteriological Examination
from those Suffering from Disease ..... 240
CHAPTER XVI.
Bacteriological Examination of Water and Air The Con-
tamination and Purification of Drinking Waters . . 245
CONTENTS i x
CHAPTER XVII.
PAGE
The Classification of Bacteria ...... 257
CHAPTER XVIII.
Bacillus of Tuberculosis (Koch's Tubercle Bacillus) . . 263
CHAPTER XIX.
Bacilli Showing Similar Staining Reactions to those of the
Tubercle Bacilli Syphilis Bacillus Smegma Bacillus
Leprosy Bacillus Grass Bacilli 311
CHAPTER XX.
Influenza Bacillus ......... 320
CHAPTER XXI.
Diphtheria Bacillus 329
CHAPTER XXII.
Tetanus Bacillus 335
CHAPTER XXIII.
Bacillus Typhosus ( Eberth-Gaff ky's Bacillus of Typhoid
Fever ; Bacillus Typhi Abdominalis) .... 402
CHAPTER XXIV.
Bacillus Coli Comnmnis (or Colon Bacillus of Escherich) . 444
CHAPTER XXV.
Pneumobacillus ( Friedlander's Bacillus) . . . 455
CHAPTER XXVI.
The Producers of Abscess, Cellulitis, Septicaemia, etc. (the
Staphylococci and Micrococcus Tetragenus) . . . 461
x CONTENTS.
CHAPTEE XXVII.
PAGE
Streptococcus Pyogenes (Streptococcus Erysipelatis ; Strep-
tococcus of Pus ; Streptococcus Pathogenes Longus) . 476
CHAPTER XXVIII.
Micrococcus Lanceolatus (Pneumococcus ; Micrococcus Pneu-
moniae Crouposae of Sternberg ; Micrococcus of Sputum
Septicaemia and Diplococcus of Fraenkel ; Diplococcus
Pneumonias of Weichselbaum ) . . . - . 498
CHAPTEE XXIX.
Diplococcus Intracellularis Meningitidis . . . . 516
CHAPTER XXX.
Micrococcus Gonorrhoaae (Gonococcus Neisser) . . . 522
CHAPTER XXXI.
Bacillus Pyocyaneus (Bacillus of Green and of Blue Pus)
Bacillus Proteus Vulgaris Bacillus of Malignant (Edema
Bacillus Aerogenes Capsulatus . .... . 535
CHAPTER XXXII.
Bacillus Anthracis (Anthrax Bacillus) Bacillus Anthracis
Symptomatici . . . 547
CHAPTER XXXIII.
Spirillum Choleras Asiaticae (Koch's Comma Bacillus of
Asiatic Cholera) 568
CHAPTER XXXIV.
Spirilla Resembling that of Cholera The Spirillum of Re-
lapsing Fever 589
CONTENTS. xi
CHAPTER XXXV.
PAGE
Glanders Bacillus 598
CHAPTER XXXVI.
Bubonic Plague Bacillus Yellow Fever Bacillus Whoop-
ing-cough Bacillus . . 606
APPENDIX.
BRIEF DESCRIPTIONS OF A FEW REPRESENTATIVE
PATHOGENIC MICRO-ORGANISMS WHICH
ARE NOT BACTERIA
CHAPTER XXXVII.
The Streptothrix Group ( Actinomyces) Favus and Ring-
worm Fungi Yeast ....... 615
CHAPTER XXXVIII.
Plasmodium Malarise (Malarial Parasites; Laverania)
Amoeba Coli (Amoeba Dysenteriae of Councilman and
Lafleur ; Dysenteric Amoeba) 626
CHAPTER XXXIX.
Vaccine The Micro-organism of Smallpox and Cowpox . 648
CHAPTER XL.
Rabies (Hydrophobia) 659
BACTERIOLOGY IN MEDICINE AND SURGERY.
INTRODUCTION.
ALTHOUGH most of the more important discoveries
in bacteriology which place it on the footing of a
science are of comparatively recent date, the founda-
tion of the study of vegetable and other micro-organisms
was laid over two centuries ago. From the earliest
times its history has been intimately associated with
that of medicine. Indeed, it is only through the inves-
tigations into the life-history of micro-organisms in their
relation to disease that our present knowledge of the
etiology, course, and prevention of the infectious diseases
has been acquired ; and it is only by the practical ap-
plication of the principles and methods of bacteriology
that many diseases can be positively diagnosed or the
problems which present themselves to the sanitarian be
certainly solved. The prominent position which bac-
teriology already holds toward medicine is, moreover,
daily increasing in importance. Original discoveries are
constantly adding to the list of known germ diseases,
and the outlook is favorable for eventually obtaining
through serums or through the toxic substances of the
micro-organisms themselves means for immunizing
against, if not curing, many of the specific infections.
Even at present bacterial products and protective
2
18 BACTERIOLOGY.
serums are used successfully as preventives in many of
the infectious diseases and as a cure in several. An
acquaintance, therefore, with the main facts and results
of bacteriology is as necessary to the education of the
modern physician as a knowledge of anatomy, pathol-
ogy, chemistry, or any of the allied sciences.
But before entering into a detailed consideration of
the subject, it may be interesting and instructive to
review briefly the most important steps which led up
to the development of the science, and upon which its
foundation rests, in which we shall see that the vast
results obtained by bacteriology were gained only
through long and laborious research, and after many
obstacles were met and overcome by indomitable per-
severance and accurate observation and experiment.
The first authentic observations of living microscopi-
cal organisms of which there is any record are those of
Athanasius Kircher, in 1671. This original investi-
gator demonstrated the presence in putrid meat, milk,
vinegar, cheese, etc., of <e minute living worms," but
did not describe their form or character.
Not long after this, in 1675, Anthony von Leeuwen-
hoek observed in rain-water putrid infusions, and in
his own and other saliva and diarrhoeal evacuations
living, motile " animalculse " of most minute dimen-
sions, which he described and illustrated by drawings.
Leeuwenhoek was a linen-draper by trade, living at
the time of his discoveries in Amsterdam, but he prac-
tised the art of lens-grinding, in which he eventually
became so proficient that he perfected a lens superior
to any magnifying glass obtainable at that day, and
with which he was enabled to see objects very much
smaller than had ever been seen before. " With the
INTRODUCTION. 19
greatest astonishment/' he writes, " I observed distrib-
uted everywhere through the material which I was
examining animalcules of the most microscopic size,
which moved themselves about very energetically."
The work of this observer is conspicuous for its purely
objective character and absence of speculation ; and his
descriptions and illustrations are done with remarka-
ble clearness and accuracy, considering the imperfect
optical instruments at his command. There is little
doubt that Leeuwenhoek really saw some of the larger
species of micro-organisms which we now recognize as
bacteria, probably spirilla.
It was not until many years later, however, that any
attempt was made to define the characters of these
minute organisms and to classify them. The first to
make such an effort was Otto Friedrich Miiller, in
1786 ; but having no means of obtaining pure cultures
all the earlier botanists naturally fell into serious errors
in the classification of bacteria. Thus various motile
organisms, which are now known to be of vegetable
origin, were commonly included under the infusoria,
which are unicellular animal organisms.
Ehrenberg, in 1838, thus describes under the gen-
eral name Vibrioniens four genera of filamentous bac-
teria :
1. Bacterium filaments linear and inflexible.
2. Vibrio filaments linear, sinuous, flexible.
3. Spirillum filaments spiral, inflexible. y.
4. Sperochcete filaments spiral, flexible.
Dujardin, in 1841, also placed the vibrioniens of
Ehrenberg among the infusoria, describing them as
extremely slender, filiform animals without appreciable
organization and without visible locomotive organs.
20 BACTERIOLOGY.
Perty, in 1852, drew attention to the vegetable
origin of these minute organisms ; Robin, in 1853,
suggested their relationship to the algae ; Davaine, in
1859, emphasized this fact ; and since it has been con-
firmed by the investigations of Cohn, Nageli, and
others. Bacteria are now generally believed by bacte-
riologists to be vegetable organisms, schizomycetes, or
fission-fungi, closely allied to the algse.
From the earliest investigations into the life-history
and properties of bacteria these micro-organisms have
been thought to play an important part in the causa-
tion of infectious diseases. The doctrine of contagium
aminatum was based upon the discoveries of Athana-
sius Kircherand Leeuwenhoek, and the " animalculse "
then observed in organic materials were believed to be
the cause of the great epidemics of the day, such as
the plague. Shortly after these first investigations,
Lange and Hauptmann advanced the opinion that
puerperal fever, measles, smallpox, typhus, pleurisy,
epilepsy, gout and many other diseases were due to
animal contagion. Andry and Linne, in 1701, as-
sumed the same cause for syphilis, and Lancisi, in
1718, for malaria. In fact, so wide- spread became the
belief in a causal relation of these minute organisms to
disease that it soon amounted to a veritable craze, and
all forms and kinds of diseases were said to be pro-
duced in this way, upon no other foundation than that
these organisms had been found in the mouth and in-
testinal contents of men and animals, and in water.
Among those who were especially conspicuous at
this time for their advanced views on the germ-theory
of infectious diseases was Marcus Antonius Plenciz, a
physician of Vienna. This acute observer, who pub-
INTRODUCTION. 21
lished his views in 1762, maintained that not only
were all infectious diseases caused by micro-organisms,
but that the infective material could be nothing else
than a living organism. On these grounds he en-
deavored to explain the variations in the period of
incubation of the different infectious diseases. He also
insisted that there were special germs for each infectious
disease by which the specific disease was produced.
Plenciz believed, moreover, that these organisms were
capable of multiplication in the body, and suggested
the possibility of their being conveyed from place to
place through the air. He also made original investi-
gations into the process of decomposition, and having
found " animalcube " in all decomposing matter, he
became so thoroughly convinced of the causative rela-
tion of these organisms to the process that he formu-
lated the law that decomposition takes place by means
of living organisms, and is possible only through their
increase.
These views, it is true, were largely speculative, and
rested upon insufficient experiment; but they were so
plausible, and the arguments put forward in their sup-
port were so logical and convincing, that they continued
to gain ground, in spite of considerable opposition and
ridicule, and in many instances the conclusions reached
have since been proved to be correct. The fact that
infectious diseases were of sudden occurrence, breaking
out often in isolated places, and that they frequently
remained clinging for long periods to certain localities,
leaving others unaffected, was evidence that they were
not produced by a gaseous infective agent. Moreover,
the mode of infection, its unlimited development among
large numbers of individuals, and gradual spread over
22 BACTERIOLOGY.
wide areas the incubation, course of, and resulting
immunity in recovery from infectious diseases all
pointed to the probable cause being a living organism.
Amoug other distinguished men of the day whose
observations exerted a most powerful influence upon
the doctrine of infection may be mentioned Henle.
His writings (Pathological Investigations, 1840, and
Text-book of Rational Pathology, 1853), in which he
described the relation of micro-organisms to infectious
diseases, and defined the character and action of bac-
teria upon certain phases and symptoms of these affec-
tions, are remarkable for their clearness and precision.
But, meanwhile, the question which most interested
these investigators into the cause of infectious diseases
was, Whence are these micro-organisms derived which
were supposed to produce them ? Were they the result
of spontaneous generation due to vegetative changes in
the substances in which the organisms were found, or
were they reproduced from similar pre-existing organ-
ismsthe so-called vitalistic theory ? This question is
intimately connected with the investigations into the
origin and nature of fermentation and putrefaction.
Among those who advocated the theory of spon-
taneous generation was Neidham, who, in 1749, at-
tempted to prove by experiment the truth of his opin-
ions. He placed a grain of barley in a watch-glass
containing water, covered it carefully, and allowed it
to germinate. On later examination he found bacteria
present, which he maintained were the result of changes
in the grain itself due to its germination.
In 1769, Spallanzani showed by another experiment
that the theory of spontaneous generation was incor-
rect. He demonstrated that if putrescible infusions
INTR OD UCTION. 23
of organic matter were placed in symmetrically sealed
flasks and then boiled the liquids were sterilized ;
neither were living organisms found in the solutions,
nor did they decompose ; and the infusions remained
unchanged for an indefinite period.
It was objected to these experiments that the high
temperature to which the liquids had been subjected so
altered them that spontaneous generation could no
longer take place. This objection was met by Spall-
anzani by cracking one of the flasks and allowing air
to enter, when living organisms and decomposition
again appeared in the boiled infusions.
Another objection raised was that in excluding the
oxygen of the air by hermetically sealing the flasks the
essential condition for the development of fermentation,
which required free admission of this gas, was inter-
fered with. This objection was then met by Schulze,
in 1836, by causing the air admitted to the boiled
decomposable liquids to pass through strong sulphuric
acid. Air thus robbed of its living organisms did not
produce decomposition ; whereas when no such precau-
tions were taken with the air admitted the boiled solu-
tions quickly fell into putrefaction, and living organ-
isms were found to be present.
Schwann, in 1839, obtained similar results in another
way; he deprived the air admitted to his boiled liquids
of micro-organisms by passing it through a tube
which was heated to a temperature high enough to
destroy them. To this investigator is also due the
credit of having discovered the specific cause the yeast
plant, or saccharomyces cerevisice of alcoholic fermen-
tation, the process by which sugar is decomposed into
alcohol and carbonic acid.
24 BACTERIOLOGY.
Helmholtz, in 1843, repeated and confirmed
Schwann's experiments with calcined air. He found
that the free admission of air so treated to boiled
organic infusions was not capable of producing fer-
mentation of any kind.
Again, it was objected to these experiments that the
heating of the air had perhaps brought about some
chemical change which hindered the production of fer-
mentation. Schroeder and von Dusch, in 1854, then
showed that by a simple process of filtration, which
has since proved of inestimable value in bacteriological
work, the air can be mechanically freed from germs.
By placing in the mouth of the flask containing the
boiled solutions a loose plug of cotton, through which
the air could freely circulate, it was found that all
suspended micro-organisms could be excluded, and
that air passed through such a filter, whether hot or
cold, did not cause fermentation of boiled infusions.
Similar results were obtained by Hoffmann in 1860,
and by Chevreul and Pasteur in 1861, without a cotton
filter, by drawing out the neck of the flask to a fine
tube and turning it downward, leaving the mouth open.
In this case the force of gravity prevents the suspended
bacteria from ascending, and there is no current of air
to carry them upward through the tube into the flask
containing the boiled infusion.
Tyndall later showed (1876), by his well-known in-
vestigations upon the floating matters of the air, that
in a closed chamber, in which the air is not disturbed
by currents, all suspended particles settle to the bottom,
the superincumbent air being optically pure, as is
proved by passing a ray of light through it. He dem-
onstrated that the presence of living organisms in
INTR OD UCTION. 25
decomposing fluids was always to be explained either
by the pre-existence of similar living forms in the in-
fusion or upon the walls of the vessel containing it, or
by the infusion having been exposed to air which was
contaminated with organisms.
These facts have since been practically confirmed on
an enormous scale in the preservation of food by the
process of sterilization. Indeed, there is scarcely any
biological problem which has been so satisfactorily
solved or in which such uniform results have been ob-
tained ; but all through the experiments of the earlier
investigators irregularities were constantly appearing.
Although in the large majority of cases it was found
possible to keep boiled organic liquids sterile in flasks
to which the oxygen of the air had free access, the
question of spontaneous generation still remained un-
settled, inasmuch as occasionally, even under the most
careful precautions, decomposition did occur in such
boiled liquids.
This fact was explained by Pasteur in I860 by ex-
periments showing that the temperature of boiling
water was not sufficient to destroy all living organ-
isms, and that, especially in alkaline liquids, a higher
temperature was required to insure sterilization. He
showed that at a temperature of 110 to 112 C. (230
to 233 F.), however, which he obtained by boiling
under a pressure of one and one-half atmospheres, all
living organisms were invariably killed.
Pasteur at a later date (1865) demonstrated that the
organisms which resist the boiling temperature are, in
fact, reproductive bodies, which he described under the
name of " corpuscles ovoides " or " corpuscles bril-
lants " now known as spores. Perty, in 1852, and
26 BACTERIOLOGY.
Robin, in 1853, had observed these highly refractile
bodies ; but it was not until 1876 that the development
of spores was carefully investigated and explained by
Cohn and later by Koch. These observers showed that
certain rod-shaped organisms possess the power of pass-
ing into a resting or spore-stage under peculiar conditions
of growth, and when in this stage they are much less
susceptible to the injurious action of higher tempera-
tures than when in their normal vegelative condition.
With this discovery the controversy of spontaneous
generation was finally settled. If these micro-organ-
isms, some of them being capable of producing the
more resistant spores, were present in the air, dust,
soil, water, etc., it was easy enough to explain the
irregularities in the foregoing experiments ; nor was it
any longer to be doubted that these bacteria, through
their products, were the cause, not the effect, of fer-
mentation and putrefaction, and that when organic sub-
stances were completely sterlized and protected against
the entrance of living germs from without, no develop-
ment of micro-organisms occurred in them.
Stimulated by the establishment of the fact that fer-
mentation and putrefaction were due to the action of
living organisms reproduced from similar pre-existing
forms, the study of the causal relation of these micro-
organisms to disease was taken up with renewed vigor.
Reference has already been made to the opinions and
hypothesis of the earlier observers as to the microbic
origin of infectious diseases. The first positive grounds,
however, for this doctrine, founded upon actual ex-
periment, were the investigations into the cause of cer-
tain infectious diseases in insects and plants. Thus
Bassi, in 1837, demonstrated that a fatal infectious
INTRODUCTION. 27
malady of the silkworm muscardine was due to a