PTOMAINES, LEUCOMAINES, 
AND 
BACTERIAL PROTEIDS: 
OR 
THE CHEMICAL FACTORS IN THE CAUSATION OF DISEASE. 
BY 
VICTOR C. VAUGHAN, Ph.D., M.D., 
PROFESSOR OF HYGIENE AND PHYSIoWlfelOAL CHEMISTRY IN THE UNIVERSITY OF MICHIGAN, 
AND DIRECTOR OF THE HYGIENIC LABORATORY ; 
AND 
FREDERICK G. NOVY, Sc.D, M.D, 
ASSISTANT PROFESSOR OF HYGIENE AND PHYSIOLOGICAL CHEMISTRY IN THE UNIVERSITY 
OF MICHIGAN. 
SECOND EDITION, REVISED AND ENLARGED. 
PHILADELPHIA: 
LEA BROTHERS & CO., 
18 91. Entered according to Act of Congress in the year 1891, by 
LEA BROTHERS & CO., 
In the Office of the Librarian of Congress at Washington, D. C. 
DOBKAS, PRINTER, 
PHILADELPHIA, TO 
ALBERT B. PRESCOTT, Ph.D., M.D., F.C.S., 
DIRECTOR OF THE CHEMICAL LABORATORY IN THE UNIVERSITY OF MICHIGAN, 
THIS LITTLE WORK 
IS RESPECTFULLY DEDICATED 
AS A SLIGHT TOKEN OF THE HIGH ESTEEM IN WHICH 
HE IS HELD BY HIS FORMER STUDENTS, 
THE AUTHORS. P It E FACE TO SECOND EDITION. 
Tiie preparation of this edition lias been made a work 
of pleasure on account of the many kind words which have 
been said concerning our first effort to collect the scattered 
facts pertaining to the chemical factors in the causation of 
disease. We must be allowed to express our gratification 
at the general acceptance accorded to the statements which 
we first made three years ago, and which were then re- 
garded by many as extremely radical. At that time 
many of the leading bacteriologists held to the “mechan- 
ical interference” theory, and regarded the chemical pro- 
ducts of germs as of some interest, but in no direct way 
concerned in the causation of disease. Now the fact that 
a germ is pathogenic is considered to be sufficient evidence 
that it elaborates poisonous products, and the study of these 
products is regarded as of the greatest importance in the 
investigation of the germ and the disease which it causes. 
The interest in this subject is not confined to a study of 
the causation of disease, but efforts are being made to 
secure immunity from disease and even to effect cures by 
the employment of the bacterial products. This line of VI 
PREFACE TO SECOND EDITION. 
study has certainly become one of great interest to all 
scientific students of medicine. 
In the preparation of the present edition we have 
endeavored to utilize the latest and best information, and 
we can only express our thanks for the encouragement 
which we have received from so many sources and hope 
that the present effort will justify no censure. 
University of Michigan, September, 1891. PREFACE TO FIRST EDITION. 
Within the past ten years much has been said and 
written concerning the basic substances formed during the 
putrefaction of organic matter, and those which are pro- 
duced by the normal tissue-changes in the living organism. 
Many investigators have given their whole time and atten- 
tion to the study of these substances, and important discov- 
eries have been made and much light has been thrown upon 
what have heretofore been considered problems in medical 
science. To collect, arrange, and systematize the facts 
concerning ptomaines and leueomames has been our first 
object. Although many short essays, some of them of 
great value, have been written with the above-mentioned 
object in view, the present work may be regarded as the 
first attempt to make this collation embrace everything of 
importance on this subject. In endeavoring to accomplish 
this object we have met with many difficulties. The original 
reports of the various investigators are scattered through 
the pages of medical and scientific journals, transactions of 
societies, monographs, government reports, etc. However, 
with few exceptions we have been able to obtain the original VIII 
PREFACE TO FIRST EDITION. 
reports, and we think that we have included everything of 
importance published up to the present year (1888). 
To the physician the facts which have been made known 
concerning the putrefactive and physiological alkaloids 
must be of great value, and if this little work furnishes 
the means by which members of the profession may become 
better acquainted with the nature of those poisons which 
are introduced from without, and those which are gener- 
ated within the body of man, the object of its authors will 
be accomplished. 
University of Michigan, July, 1888. CONTENTS. 
PAGE 
Introduction 13 
CHAPTER I. 
Definition and Classification of the Bacterial 
Poisons 15 
CHAPTER H. 
Historical Sketch of the Bacterial Poisons . . 22 
CHAPTER IN. 
Foods Containing Bacterial Poisons : Poisonous Mus- 
sels, Oysters and Eels, Fish, Sausage, Ham, Canned 
Meats, Cheese, Milk, Ice-cream, Meal and Bread . . 36 
CHAPTER IV. 
General Considerations of the Relation of Bac- 
terial Poisons to Infectious Diseases : Classifica- 
tion of Diseases, How Germs Produce Disease, Definition 
of Infectious Disease, Koch’s Rules ... .84 
CHAPTER V. 
The Bacterial Poisons of some of the Infectious 
Diseases : Anthrax, Asiatic Cholera, Tetanus, Tuber- 
culosis, Diphtheria, Suppuration, The Summer Diar- 
rhoeas of Infancy, Typhoid Fever, Swine-plague (Hog- 
cholera), Rabbit Septicaemia, Pneumonia, Malignant 
CEdema, Puerperal Fever 101 
CHAPTER VI. 
The Nature of Immunity-giving Substances : Methods 
of securing Immunity ; Bacterial Products which Favor 
the Development of Infectious Diseases . . . 146 X 
CONTENTS. 
chaptp;r vii. 
PAGE 
The Germicidal Proteids of the Blood . . . 152 
CHAPTER VIII. 
Methods of Extracting Ptomaines. Basic Impurities in 
Reagents. The Stas-Otto Method, Dragendorff’s Method, 
Brieger’s Method, The Methods of Gautier and Etard. 
Remarks upon the Methods 157 
CHAPTER IX. 
Methods of Isolating the Bacterial Proteids . . 171 
CHAPTER X. 
The Importance of Ptomaines to the Toxicologist. 
Coniine-like Substances, Nicotine, Strychnine, Mor- 
phine, Atropine, Digitaline, Veratrine, Delphinine, Col- 
chicine. Effect of Ptomaines on Alkaloidal Reactions . 174 
CHAPTER XI. 
Chemistry of the Ptomaines: Primary Amines, Dia- 
mines, etc., the Choline Group, other Oxygen-containing 
Bases, Undetermined Ptomaines. Tables . . .187 
CHAPTER XII. 
Chemistry of the Leucomaines: Uric Acid Group, 
Creatinine Group, Undetermined Leucomaines. Tables 280 
CHAPTER XIII. 
The Autogenous Diseases 352 
CHAPTER XIV. 
Bibliography: Ptomaines, Leucomaines, Bacterial Pro- 
teids, Miscellaneous 364 PTOMAINES, LEUC0MA1NES, AND BACTERIAL 
PROTEIDS. 
INTRODUCTION. 
It is customary to divide bacteria into the parasitic and 
the saprophytic. The obligate parasite can live only on 
living matter; the obligate saprophyte can live only on 
dead matter. Since all attempts to grow the bacilli of 
syphilis and leprosy on artificial media have failed, they are 
probably obligate parasites. True parasitic germs do not 
prove speedily fatal to their hosts, because their continued 
existence depends upon the continued existence of their host, 
or on their transference to another host. Leaving out of 
consideration the obligate bacterial parasites, about which 
very little is known at best, the above classification becomes 
of but little importance to us in a study of the causal rela- 
tion of germs to disease, because a given bacterium may 
grow and multiply in one part of the body, while it is 
unable to do so in another; or it may thrive in one species 
of animal, while it finds the conditions unfavorable in an- 
other species ; or similar differences may exist in individual 
members of the same species. Thus, the white rat is ordi- 
narily and naturally immune against the bacillus of anthrax, 
but if the rat be exhausted by being kept on a small tread- 
mill for some hours it becomes susceptible to anthrax. 
Recognizing these facts, we propose that bacteria be divided 
into the toxicogenic and the non-toxicogenic. Since we 
know of no infectious disease in which poisons are not 
formed, the toxicogenic germs only are of interest to us. 14 
INTRODUCTION. 
Iii the study of these we must not only ascertain the nature 
of the poisons which they produce, but must know the con- 
ditions under which they can multiply and elaborate these 
poisons. To these points the following pages, in so far as 
they treat of the infectious diseases, will be devoted. 
However, all diseases are not infectious; all poisons 
formed within the body do not owe their existence to bac- 
teria. Some originate in the altered metabolism of the 
various tissues, and these will be discussed under the auto- 
genous diseases. CHAPTER I. 
DEFINITION AND CLASSIFICATION OF THE BACTERIAL 
POISONS. 
Ptomaines.—An exact classification of the chemical 
factors in the causation of the infectious diseases can prob- 
ably not be made at present. We know of two chemically 
distinct classes, one of which contains substances which 
combine with acids, forming chemical salts, and which in 
this respect at least correspond with the inorganic and 
vegetable bases. The members of this class are designated 
as ptomaines, a name suggested by the Italian toxicologist, 
Selmi, and derived from the Greek word mo/ia, meaning a 
cadaver. A ptomaine may be defined as a chemical com- 
pound which is basic in character and which is formed by 
the action of bacteria on organic matter. On account of 
their basic properties, in which they resemble the vegetable 
alkaloids, ptomaines may be called putrefactive alkaloids. 
They have also been called animal alkaloids, but this is a 
misnomer, because, in the first place, some of them are 
formed in the putrefaction of vegetable matter; amt, in 
the second place, the term “animal alkaloid” is more prop- 
erly restricted to the leucomaines—those basic substances 
which result from tissue metabolism in the body. While 
some of the ptomaines are highly poisonous, this is not an 
essential property, and others are wholly inert. Indeed, 
the greater number of those which have been isolated up 
to the present time do not, when employed in single doses, 
produce any apparently harmful effects. Brieger restricts 
the term ptomaine to the non-poisonous basic products, and 
designates the poisonous ones as “ toxines.” This is a 
classification, however, which seems to be of questionable 
utility. It is not always easy to say just what bodies are 
poisonous and what are not. The poisonous action of a 16 
PTOMAINES. 
substance depends upon the conditions under which, and 
the time during which, it is administered. Thirty grains 
of quinine may be taken by a healthy man during twenty- 
four hours without any appreciably ill effect, yet few of us 
would be willing to admit that the administration of this 
amount daily for three months would be wise or altogether 
free from injury. In the same manner the administration 
of a given quantity of a putrefactive alkaloid to a dog or 
guinea-pig in a single dose may do no harm, while the 
daily production of the same substance in the intestine of 
a man and its absorption continued through weeks and pos- 
sibly months may be of marked detriment to the health. 
We do not as yet know enough about the physiological or 
toxicological action of the putrefactive alkaloids to render 
the classification proposed by Brieger worthy of general 
adoption. 
All ptomaines contain nitrogen as an essential part of 
their basic character. In this they resemble the vegetable 
alkaloids. Some of them contain oxygen, while others do 
not. The latter correspond to the volatile vegetable alka- 
loids, nicotine and coniine, and the former correspond to 
the fixed alkaloids. 
Since all putrefaction is due to the action of bacteria, it 
follows that all ptomaines result from the growth of these 
microorganisms. The kind of ptomaine formed will de- 
pend upon the individual bacterium engaged in its produc- 
tion, the nature of the material being acted upon, and the 
conditions under which the putrefaction goes on, such as the 
temperature, amount of oxygen present, and the duration 
of the process. 
Brieger found that, although the Eberth bacillus grew 
well in solutions of peptone, it did not produce any pto- 
maine; while from cultures of the same bacillus in beef-tea 
he obtained a poisonous alkaloid. Fitz found that whilst 
the bacillus butyricus produces by its action on carbohy- 
drates butyric acid, in glycerin it produces propylic alcohol, 
and Morin has found amyl alcohol among the products of 
this germ. Brown has shown that while the inycoderma 
aceti converts ethyl ic alcohol into acetic acid, it converts 17 
DEFINITION. 
p ropy lie alcohol into propionic acid, and is without effect 
upon methylic alcohol, primary isobutylic alcohol, and 
ainylic alcohol. Some bacteria Avill not multiply below a 
given temperature. Thus, the bacillus butyricus will not 
grow at a temperature below 24°.1 The lower temperature 
does not destroy the organism, but it lies dormant until the 
conditions are more favorable for its growth. Pasteur 
divided the bacteria into two classes—the aerobic and the 
anaerobic. As the name implies, the former grow and 
thrive in the presence of air, while the latter find their 
conditions of life improved by the exclusion of air. There- 
fore, different ptomaines will be formed in decomposing 
matter freely exposed to the air, and in that which is buried 
beneath the soil or from which the air is largely excluded. 
Even when the same ferment is present the products of the 
putrefaction will vary, within certain limits, according to 
the extent to which the putrefying material is supplied with 
air. The kind of ptomaine found in a given putrid sub- 
stance will depend also upon the stage of the putrefaction. 
Ptomaines are transition products in the process of putre- 
faction. They are temporary forms through which matter 
passes while it is being transformed, by the activity of bac- 
terial life, from the organic to the inorganic state. Com- 
plex organic substances, as muscle and brain, are broken 
up into less complex molecules, and so the process of 
chemical division goes on until the simple and well-known 
final products, carbonic acid gas, ammonia, and water, 
result; but the variety of combinations into which an 
individual atom of carbon may enter during this long 
series of changes is almost unlimited, and with each change 
in combination there is more or less change in nature. In 
one combination the atom of carbon may exist as a con- 
stituent of a highly poisonous substance, while the next 
combination into which it enters may be wholly inert. 
It was formerly supposed that putrefaction was simply 
oxidation, but the researches of Pasteur and others have 
demonstrated the fact that countless myriads of minute 
1 All temperatures given in this work are Centigrade, unless otherwise 
specified. 18 
BACTERIAL PROTEIDS. 
organisms are engaged constantly in transforming matter 
from the organic to the inorganic form. Lock up the bit 
of flesh so that these little workers cannot reach it, and it 
will remain unchanged indefinitely. 
It may be asked if any of the changes occurring during 
putrefaction are to be regarded as purely chemical. Without 
doubt, many of the secondary products of putrefaction arise 
from reactions between antecedent and more complex prod- 
ucts or by the action of oxygen, water, and reducing agents 
upon primary products. Ptomaines formed in this way 
may be regarded as the indirect results of bacterial life. 
Bacterial Proteids.—These substances have been 
known for so short a time and are at present so imperfectly 
known that many difficulties arise in discussing them. In 
the first place, we may divide the bacterial proteids into 
two classes : (1) those which constitute an integral part of 
the bacterial cells, and (2) those which have not been 
assimilated by the cells, but which have been formal by 
the fermentative or cleavage action of the bacteria on the 
proteid bodies in which they are growing. Even this 
classification is of questionable value. We allow bacteria 
to grow for a number of days in a nutrient solution. We 
then separate the soluble constituents from the formal cells 
by filtration through porous tile; we wash the latter and 
then study their proteid contents, which constitute the first 
class, as given above ; but the filtrate contains, or may con- 
tain, any one or more of the following proteid bodies : (1) 
Those portions of the proteid substances which were used 
in the preparation of the nutrient solution and which have 
escaped the action of the bateria ; (2) proteids which have 
been at one time integral parts of the cells, but which 
have passed into solution on the death and dissolution of 
the bacteria; and (3) proteids which have been formed by 
the fermentative action of the bacteria, or those which are 
defined as constituting the second class, as given above. 
We know at present of no means by which one of these 
proteids can with certainty be isolated from the others. 
However, the above classification is a convenient one, and BACTERIAL proteids. 
19 
with a clear understanding that it is not free from criticism 
we may employ it until a more thorough and scientific study 
of these bodies has been made. 
The difficulty in discussing these substances lies not only 
in the classification, but in the name which shall be em- 
ployed to designate them. Brieger and Franker have 
proposed the term “ toxalbuminsbut, while it is true 
that some belong to the albumins, others are more truly 
albumoses ; others are most closely related to the peptones ; 
and still others differ in some important respects from all 
of these. In view of the above facts, we have decided upon 
the term “ bacterial proteids ” to designate those formed by 
the fermentative action of germs, while those which consti- 
tute an integral part of the cell will be known as “ the 
bacterial cellular proteids.” 
The Bacterial Cellular Proteids.—Nencki first prepared 
one of these substances from putrefactive bacteria. These 
were obtained by decantation, freed from fat with ether, 
dissolved in fifty parts of a potash solution of 0.5 percent., 
heated for some hours at 100° and filtered. The filtrate 
was acidified with dilute hydrochloric acid and precipitated 
by the addition of rock salt. The precipitate was washed 
with a saturated salt solution, dried at 100°, and washed 
free from salt with water. Nencki designates this sub- 
stance as “ mycoproteiu,” and finds that it has the formula, 
C25H42N609. Freshly precipitated mycoprotein forms in 
amorphous flakes, which are soluble in water, alkalies, and 
acids. The aqueous solution is acid in reaction. After 
being dried at 100° it is no longer wholly soluble in water. 
Nencki found that it is not precipitated from aqueous solu- 
tion by alcohol, but by picric acid, tannic acid, and mercuric 
chloride; that it does not give the xanthoproteid, but 
does give the Millon and the biuret reactions. According 
to Schaffer it is changed by acids into peptone, and on 
being fused with five parts of potash it breaks up into am- 
monia, amylamin, phenol (0.15 per cent, of its weight), vale- 
rianic acid (38 per cent.), leucine, and traces of indol and 
skatol. A proteid obtained from the yeast plant has the 
formula, C12I r21N303. 20 
BACTERIAL PROTEIDS. 
The purified pyogenetic agent obtained from the pneu- 
monia bacillus of Friedrander was found by Buchner 
to give the following reactions: It is soluble in water and 
the concentrated mineral acids, very soluble in dilute alka- 
lies, from which it is precipitated on the addition of an acid. 
From its aqueous solution, it is not precipitated by heat, 
nor by saturation with sodium chloride, but is precipitated 
by magnesium sulphate, copper sulphate, platinum chloride, 
gold chloride, lead salts, picric acid, tannic acid, and abso- 
lute alcohol. It gives the xanthoproteid, Millon, and 
biuret reactions. 
The Bacterial Proteids.—Brieger and Franker ob- 
tained the proteid poison of diphtheria by precipitating 
the filtrate from a Chamberland filter after concentration 
to one-third its volume at 30°, with absolute alcohol after 
feebly acidifying with acetic acid. The precipitate was puri- 
fied by repeated solution in water and reprecipitation with 
alcohol. Dried in a vacuum at 40°, it forms a snow-white, 
amorphous, very light mass. From its aqueous solution it 
is not precipitated by heat or dilute nitric acid, singly or 
combined, nor by sodium sulphate, sodium chloride, mag- 
nesium sulphate, or lead salts. It is precipitated by car- 
bonic acid (to saturation), concentrated mineral acids, 
potassium ferrocyanide and acetic acid, phenol, organic 
acids (soluble in excess), copper sulphate, silver nitrate, 
and mercuric chloride. The so-called alkaloidal reagents, 
phosphomolybdic acid, potassio-meeuric iodide, potassio- 
bismuthic iodide, platinum chloride, gold chloride, and 
picric acid also cause precipitation. The xanthoproteid, 
Millon, and biuret reactions give positive results. An ulti- 
mate analysis furnishes the following figures computed 
from the ash-free substance : C 45.35, II 7.13, N 16.33, S 
1.39, O 29.80. From these facts Brieger and Franker 
conclude that this substance is allied to serum-albumin. 
Their bouillon cultures contain serum-albumin, and they 
suppose that the bacteria convert this into the poison by 
causing a rearrangement in the atoms; but the same poison 
was formed when nutrient solutions containing no proteid BACTERIAL PROTEIDS. 
21 
save peptone were employed. In this ease they suppose 
that the bacteria reconvert the peptone into an albumin. 
The poisonous proteids obtained by Brieger and 
Franker from cultures of the Eberth germ, the comma 
bacillus, and the staphylococcus aureus are practically in- 
soluble in water, and more nearly related to the globulins 
than the albumins, although they differ from the former 
in their tardy and difficult solubility in dilute solutions of 
sodium chloride. 
The poisonous proteids isolated by Vaughan from cul- 
tures of two species of toxieogenic germs found in drinking 
water, supposed to be the cause of typhoid fever, are solu- 
ble in water, from which they are not precipitated by boil- 
ing, or by concentrated nitric acid, or by both. Potassium 
ferrocyanide and acetic; acid, sodium sulphate, magnesium 
sulphate, and carbonic acid also fail to precipitate them. 
They are precipitated by the general alkaloidal reagents, 
and respond to the xanthoproteid, Millon, and biuret tests. 
They are precipitated by ammonium sulphate when added 
to saturation, and for this reason cannot be classed among 
the peptones. Neither benzoyl chloride nor phenyl-hydra- 
zin chloride precipitate them. Their poisonous properties 
are destroyed by prolonged boiling or by being heated to 
80° for some hours, though they remain active after an 
exposure of ten minutes to the last mentioned temperature. 
Of the three bacterial proteids obtained by the same ex- 
perimenter from the bacilli x, a and A of Booker’s list of 
summer diarrhoea germs, the first two are soluble in water, 
while the other is not. So far as their behavior with pre- 
cipitating agents is concerned, the first two agree with the 
proteids of the water germs. 
Ttzzoni and Cattani find that the proteid of cultures 
of their tetanus germ is rendered inert by precipitation with 
absolute alcohol. It is obtained by saturation with am- 
monium sulphate, and the removal of the salt by dialysis. 
Further description of the individual proteids will be 
given in subsequent chapters. CHAPTER II. 
HISTORICAL SKETCH OF THE BACTERIAL POISONS. 
It must have been known to primitive man that the 
eating of putrid flesh was liable to affect the health more 
or less seriously; and when lie began his endeavors to 
preserve his food for further use, instances of poisoning 
from putrefaction must have multiplied. However, the 
distinguished physiologist, Albert von Haller, seems 
to have been the first to make any scientific experiments 
concerning the effects of putrid matter upon animals. He 
injected aqueous extracts of putrid material into the veins 
and found that death resulted. Later in the eighteenth 
century Morand gave an account of the symptoms in- 
duced by eating poisonous meat. In the early part of 
the present century (1808 to 1814) Gaspard carried on 
similar experiments. He use as material the putrid flesh 
of both carnivorous and herbivorous animals. With these 
he induced marked nervous disturbances, as stiffness of the 
limbs, opisthotonos, and tetanus. Gaspard concluded from 
the symptoms that the poisonous effects were not due to 
carbonic acid gas or hydrogen sulphide, but thought it 
possible that ammonia might have part in their produc- 
tion. In 1820 Kerner published his first essay on poi- 
sonous sausage, which was followed by a second in 1822. 
At first he thought that the poisonous properties were due 
to a fatty acid, similar to the sebacic of Thenard, and 
which originated during putrefaction. Later he modified 
these views, and believed the pron to be a compound con- 
sisting of the sebacic acid auttavolmrfe principle. This may 
be regarded as the first suggestion as to the probability of 
the development of a poisonous substance with basic prop- 
erties in decomposing matter. In 1822, Dupre observed 
a peculiar disease among the soldiers under his care, who, historical sketch. 
23 
during the warm and dry summer of that year, were 
compelled to drink very foul water. Later Magendie, 
induced by the investigations of Gaspard and the obser- 
vations of Dupre, made many experiments, in which dogs 
and other animals were confined over vessels containing 
putrid animal matter and compelled constantly to breathe 
the emanations therefrom.. The effects varied markedly 
with the species of animal and the nature of the putrid 
material, but in some instances symptoms were induced 
which resembled closely those of typhoid fever in man. 
Leuret directed his attention to the chemical changes 
produced in blood by putrefaction, but accomplished noth- 
ing of special value. Dupuy injected putrid material into 
the jugular vein of a horse, and with Trousseau studied 
alterations produced in the blood by these injections. 
During the third decade of the present century there 
were many investigators in addition to those mentioned 
above, who endeavored to ascertain the active agent in 
poisonous foods. Dann, Weiss, Buchner, Schumann, 
Cadet de Gassicourt, and Or Fi r, a studied poisonous 
sausage, but made no advance upon the work done by 
Keener. Henneman, Hunnefelp, Westrumb, and 
Serturner made contributions concerning poisonous 
cheese, but all believed the caseic acid of Keener to 
be the poisonous principle. 
In 1850 Schmidt, of Dorpat, made some investigations 
on the decomposition products and volatile substances 
found in cholera stools; and, two years later, Meyer, of 
Berlin, injected the blood and stools of cholera patients 
into lower animals. In 1853 Stich made an important 
contribution on the effects of acute poisoning with putrid 
material. He ascertained that, when given in sufficient 
quantity, putrid matter produces an intestinal catarrh, with 
choleraic stools. Nervous symptoms, trembling, unsteady 
gait, and, finally, convulsions were also observed. Stich 
made careful post-mortem examinations, and was unable 
to find any characteristic or important lesions. Theo- 
retically, lit! concluded that the putrid material contained 
a ferment which produced rapid decomposition of the blood. 24 
BACTERIAL TOISONS. 
Iii 1856 Panum published a most important contribu- 
tion to the knowledge of the nature of the poison present 
in putrid flesh. He first demonstrated positively the 
chemical character of the poison, inasmuch as he showed 
that the aqueous extract of the putrid material retained 
its poisonous properties after treatment which would insure 
the destruction of all organisms, Ilis conclusions were as 
follows: 
(1) “The putrid poison contained in the decomposed 
flesh of the dog, and which is obtained by extraction with 
distilled water and repeated filtration, is not volatile, but 
fixed. It does not pass over on distillation, but remains in 
the retort. 
(2) “The putrid poison is not destroyed by boiling, nor 
by evaporation. It preserves its poisonous properties even 
after the boiling has been continued for eleven hours, and 
after the evaporation has been carried to complete desicca- 
tion at 100°. 
(3) “The putrid poison is insoluble in absolute alcohol, 
but is soluble in water, and is contained in the aqueous ex- 
tract which is formed by treating with distilled water the 
putrid material which has previously been dried by heat 
and washed with alcohol. 
(4) “The albuminoid substances which frequently are 
found in putrid fluids are not in themselves poisonous only 
so far as they contain the putrid poison fixed and condensed 
upon their surfaces, from which it can be removed bv 
repeated and careful washing. 
(5) “The intensity of the putrid poison is comparable 
to that of the venom of serpents, of curare, and of certain 
vegetable alkaloids, inasmuch as 0.012 of a gramme of the 
poison, obtained by extracting with distilled water putrid 
material which had been previously boiled for a long time, 
dried at 100°, and submitted to the action of absolute 
alcohol, was sufficient almost to kill a small dog.” 
Panum made intravenous injections with this poison, and 
with ammonium carbonate, ammonium butyrate, ammo- 
nium valerianate, tyrosine, and leucine, and found that the 
symptoms induced by the putrid poison differed from those HISTORICAL SKETCH. 
25 
caused by the other agents. Moreover, he found the symp- 
toms to differ from those of typhoid fever, cholera, pyaemia, 
anthrax, and sausage poisoning. He was also in doubt as to 
whether the poison acted directly upon the nervous system, 
or whether it acted as a ferment upon the blood, causing 
decomposition, the products of which affected the nerve- 
centres ; but he was sure that it could not correspond to 
the ordinary ferments, inasmuch as it was not decomposed 
by prolonged boiling nor by treatment with absolute alcohol. 
Certainly, the putrid poison could not consist of a living 
organism. 
The symptoms observed by Panum varied greatly with 
the quantity of the poison used and the strength of the 
animal. After the intravenous injection of large doses, 
death followed in a very short time. In these eases there 
were violent cramps, and involuntary evacuations of the 
urine and feces; the respirations were labored, the pallor 
was marked, sometimes followed by cyanosis, the pulse 
feeble, the pupils widely dilated, and the eyes projecting. 
In these eases the autopsy did not reveal any lesion, save 
that the blood was dark, imperfectly coagulated and slightly 
infiltrated through the tissue. Post-mortem putrefaction 
came on with extraordinary rapidity. 
When smaller doses or more vigorous animals were used, 
the symptoms did not appear before from a quarter of an 
hour to two hours, and sometimes even later. In these 
eases the symptoms were less violent, and the animal gen- 
erally recovered. In all instances, however, the disturbances 
were more or less marked. 
In addition to the “ putrid poison,” Panum obtained a 
narcotic substance, the two being separated by the solubility 
of the narcotic in alcohol. The alcoholic extract was evap- 
orated to dryness, the residue dissolved in water and injected 
into the jugular vein of a dog. The animal fell into a deep 
sleep, which remained unbroken for twenty-four hours, 
when it awoke apparently in perfect health. 
Panum’s first contributions, which were published in 
Danish, did not attract the attention which they deserved, 
until after the lapse of several years. Now, however, their 26 
BACTERIAL POISONS. 
importance is fully appreciated, and the distinguished inves- 
tigator lived to receive the credit and honor due him. 
Weber, in 1804, and Hemmer and Schwenninger 
in 1866, confirmed the results obtained by Panum ; and 
Schwenninger announced that in the various stages of 
putrefaction different products are formed, and that these 
vary in their effects upon animals. In 1866, Pence 
Jones and Dupre obtained from the liver a substance 
which in solutions of dilute sulphuric acid gives the blue 
fluorescence observed in similar solutions of quinine. To 
this substance they gave the name “animal chinoidine.” 
Subsequently, the same investigators found this substance 
in all organs and tissues of the body, but most abundantly 
in the nerves. Its feebly acid solutions give precipitates 
with iodine, potassio-mercuric iodide, phospho-molybdic 
acid, gold chloride, and platinum chloride. From three 
pounds of sheep’s liver, they obtained three grammes of a 
solution in which, after slight acidulation with sulphuric 
acid, the intensity of the fluorescence was about the same 
as that of a similarly acidulated solution of quinine sulphate 
which contained 0.2 gramme of quinine per litre. Still 
later, this base was obtained bv Marino-Zuco. 
In 1868, Bergmann and Sciimiedeberg separated, 
first from putrid yeast, and subsequently from decomposed 
blood, in the form of a sulphate, a poisonous substance 
which they named sepsine. The sulphate of sepsine forms 
in needle-shaped crystals. Small doses (0.01 gramme) of 
this substance were dissolved in water and injected into the 
veins of two dogs. In a short time it produced vomiting, 
and later diarrhoea, which, in one of the animals, after a 
time, became bloody. Post-mortem examination showed, 
in the stomach and intestines, bloody ecchymoses. It was 
now believed that the “putrid poison ” of Panum had been 
isolated, and that it was identical with sepsine, but further 
investigations showed that this was not true. There are 
marked differences in their effects upon animals, and sepsine 
has not been found to be generally present in putrid ma- 
terial. It is only rarely found in blood, and the closest 
search has failed to show its presence in pus. Bergmann, HISTORICAL SKETCH. 
27 
following the same method which he had used in extracting 
this poison from yeast, has been unable to obtain it from 
other putrid material. Moreover, he was not always suc- 
cessful in obtaining the poison from yeast. Sepsine was 
not obtained in quantity sufficient to serve for an ultimate 
analysis, hence, its composition remains unknown. 
In 1869 ZtiLZER and Sonnenschein prepared from 
decomposed meat extracts a nitrogenous base, which in its 
chemical reactions and physiological effects resembled atro- 
pine and hyoscyamine. When injected under the skin of 
animals it produced dilatation of the pupils, paralysis of 
the muscles of the intestines, and acceleration of the heart- 
beat ; but it is uncertain and inconstant in its action. This 
probably results from rapid decomposition taking place in 
it, or to variations in its composition at different stages of 
putrefaction. This substance has also been obtained from 
the bodies of those who have died from typhoid fever, and 
it may be possible that the belladonna-like delirium which 
frequently characterizes the later stages of this disease is 
due to the ante-mortem generation of this poison within 
the body. 
Since 1870 many chemists have been engaged in making 
investigations on the products of putrefaction. We can 
only mention a few names at present, while others will be 
referred to subsequently in discussing the individual pto- 
maines. 
First of all stands the Italian Selmi, who suggested the 
name ptomaine, and whose researches furnished us with 
much information of value, and, what is probably of more 
importance, gave an impetus to the study of the chemistry 
of putrefaction, which has already been productive of much 
good and gives promise of much more in the future. Selmi 
showed that ptomaines could be obtained (1) by extracting 
acidified solutions of putrid material with ether; (2) by 
extracting alkaline solutions with ether; (3) by extracting 
alkaline solutions with chloroform ; (4) by extracting with 
amylic alcohol ; and (5) that there yet remained in the solu- 
tions of putrid matter ptomaines which were not extracted 
by any of the above-mentioned reagents. In this way he 28 
BACTERIAL POISONS. 
gave some idea of the great number of alkaloidal bodies 
which might be formed among the products of putrefaction, 
and the promising field thus discovered and outlined was 
soon occupied by a busy host of chemists. In the second 
place, he demonstrated the fact that many of the ptomaines 
give reactions similar to those given by the vegetable alka- 
loids. This led the toxicologist into investigations, the 
results of some of which we will ascertain further on. 
Selmi, however, did not succeed in isolating completely 
a single putrefactive alkaloid. All his work was done with 
extracts. lie remained ignorant, except in a general way, 
of the composition of these bodies. Nencki, in 187fi, 
made the first ultimate analysis and determined the first 
formula of a ptomaine. This was an isomer of collodine, 
which will be described later. 
ftoRSCH and Fassbender, in a case of suspected poison- 
ing, obtained by the Stas-Otto method a liquid which 
could be extracted from acid as well as alkaline solutions 
by ether, and which gave all the general alkaloidal reac- 
tions. They were unable to crystallize either extract by 
taking it up with alcohol and evaporating. The colorless 
aqueous solution was not at all bitter to the taste. The 
precipitate formed with phospho-molybdic acid dissolved on 
the application of heat, giving a green solution, which 
became blue on the addition of ammonia. They believed 
that this substance was derived from the liver, since fresh 
ox-liver, treated in the same manner, gave them an alkaloid 
which could be extracted with ether from acid as well as 
from alkaline solutions. Gunning found this same alka- 
loid in liver-sausage from which poisoning had occurred. 
Rorsch and Fassbender state that while in some of its 
reactions this substance resembles digitaline, it is distin- 
guished from this vegetable alkaloid by the failure of the 
ptomaine to give the characteristic bitter taste. 
Schivanert, whilst examining the decomposing intes- 
tines, liver, and spleen of a child which had dic'd suddenly, 
perceived a peculiar odor and obtained by the Stas-Otto 
method (ether extract from an alkaline solution) small 
quantities of a base, which was distinguished from nicotine HISTORICAL SKETCH. 
29 
and coniine by its greater volatility and its peculiar odor. 
He supposed that this substance was produced by decom- 
position, and, in order to ascertain the truth of his suppo- 
sition, he took the organs of a cadaver that had lain for 
sixteen days at a temperature of 30° and was well decom- 
posed. These were treated with tartaric acid and alcohol. 
The acid solution was first extracted with ether, and yielded 
no result; it was then rendered alkaline and extracted 
with ether. The latter extract gave, on evaporation, the 
same substance which he had found in the organs of the 
child. The residue was a yellowish oil, having an odor 
somewhat similar to propylamine. It was repulsive, but 
not bitter to the taste, and alkaline in reaction. On the 
addition of hydrochloric acid, it crystallized in white needles, 
which were freely soluble in water, but soluble with diffi- 
culty in alcohol. On the addition of ammonium hydrate 
to this crystalline substance, a white vapor of unpleasant 
odor was given off. The crystals dissolved in sulphuric acid, 
forming a solution which was at first colorless, but which 
gradually became dirty brownish-yellow, and grayish- 
brown on the application of heat. On being warmed with 
sodium molybdate, a splendid blue color, becoming gradu- 
ally gray, was produced. Potassium bichromate and sul- 
phuric acid gave a reddish-brown, then a grass-green color. 
Nitric acid gave a yellow color. A tartaric acid solution 
of the crystals produced, on the addition of platinum chlo- 
ride, a dirty yellow precipitate of small six-sided stars, 
which contained 31.55 per cent, of platinum. Gold chlo- 
ride gave a pale yellow, amorphous precipitate; mercuric 
chloride yielded white crystals ; potassio-mercuric iodide a 
dirty-white precipitate; and potass io-cadmic iodide yielded 
no result. Tannic acid produced only a turbidity. Sodium 
phospho-molybdate gave a yellow, flocculent precipitate, 
which became blue on the addition of ammonium hydrate. 
This base has a slight reducing power, and in this it 
resembles a substance obtained by Selmi, but it differs 
from Selmi’s extract inasmuch as it does not give a violet 
coloration on being warmed with sulphuric acid. In its 
amorphous character, its behavior to the general alkaloidal 80 
BACTERIAL POISONS. 
reagents, and its lack of bitter taste, it resembles the base 
obtained by Rorsch and Fassbender, but, unlike that 
alkaloid, it is extractable from alkaline solutions only. 
Selmi, iii commenting upon the base studied by Rorsch 
and Fassbender, Schwanert, and himself, believing 
that all were dealing with the same body, states that it 
does not contain phosphorus, and that it is separated with 
extreme difficulty from the vegetable alkaloids. 
Liebermann, in examining the somewhat decomposed 
stomach and intestines in a case of suspected poisoning, 
found an alkaloidal body which was unlike that studied by 
the chemists mentioned above, inasmuch as it was not vola- 
tile. The Stas-Otto method was employed. The ether 
extract from alkaline solution left, on evaporation, a brown- 
ish, resinous mass, which dissolved in water to a turbid 
solution, the cloudiness increasing on heating. This reac- 
tion agrees with coniine, but the odor differed from that of 
the vegetable alkaloid. The aqueous, strongly alkaline 
solution gave the following reactions : 
(1) With tannic acid, a white precipitate. 
(2) With potassium iodide, a yellowish-brown, turning 
to dark-brown precipitate. 
(3) With chlorine water, a marked white cloudiness. 
(4) With phospho-molybdie acid, a yellow precipitate. 
(5) With potassio-mercuric iodide, a white precipitate. 
(6) With mercuric chloride, a white cloudiness. 
(7) With concentrated sulphuric acid, after a while, a 
redd ish-v iolet coloration. 
(8) With concentrated nitric acid, after evaporation, a 
yellowish spot. 
These reactions exclude all vegetable alkaloids save 
coniine. The putrefactive alkaloid does not distil when 
heated on the oil-bath to 200°, while coniine distils at 
135°. The former is with certainty distinguished from 
coniine by its non-poisonous properties. 
This substance is extracted bv ether from acid, as well 
as from alkaline solutions. The yellow, oily drops ob- 
tained after the evaporation of the ether are soluble in 
alcohol. The taste is slightly burning. HISTORICAL SKETCH. 
31 
Selmi obtained from both putrefying and fresh intes- 
tines a substance which gave the general alkaloidal reac- 
tions with potassium iodide, gold chloride, platinum chlo- 
ride, potassio-mercuric iodide, and phospho-mofybdic acid. 
It has strong reducing power, and when warmed with 
sulphuric acid gives a violet coloration. These reactions 
are not due to leucine, tyrosine, creatine, or creatinine. 
This is the substance which, as has been stated, Selmi con- 
sidered identical with that observed by Rorsch and Fass- 
bender and Schwanert. The minor differences observed 
by the different chemists may have been due to the varying 
degrees of purity in which the substance was obtained by 
them. 
From human bodies which had been dead from one to 
ten months, Selmi removed many alkaline bases. From 
an ether solution of a number of these, one was removed 
by treatment with carbonic acid gas. One base which was 
insoluble in ether, but readily soluble in amylic alcohol, 
was found to be a violent poison, producing in rabbits 
tetanus, marked dilatation of the pupils, paralysis, and 
death. 
Parts of a human body preserved in alcohol were found 
by Selmi to yield an easily volatile, phosphorus-containing 
substance, which is soluble in ether and carbon disulphide, 
and gives a brown precipitate with silver nitrate. It is 
not the phosphide of hydrogen. A similar substance is 
produced by the slow decomposition of the yolks of eggs. 
With potassium hydrate it gives oft" ammonia and yields a 
substance having an intense coniine odor. It is volatile 
and reduces phosphomolybdic acid. 
Selmi also obtained from decomposing egg-albumin a 
body, whose chloride forms in needles, and which has a 
curare-like action on frogs. From one arsenical body which 
had been buried for fourteen days, he obtained, by extract- 
ing from an alkaline (made alkaline with baryta) solution 
with ether, a substance which formed in needles and which 
gave crystalline salts with acids. With sulphuric acid it 
gave a red color; with iodic acid and sulphuric acid it 
liberated free iodine and gave a violet coloration; with 32 
BACTERIAL POISONS. 
nitric acid it gave a beautiful yellow, which deepened on 
the addition of caustic potash. Platinum chloride gave no 
precipitate save in highly concentrated solutions. From a 
second arsenical body, Selmi obtained by the same method 
a substance which gave, with tannic acid, a white precipi- 
tate; with iodine in hydriodic acid a kermes-brown; with 
gold chloride a yellow, which was soon reduced ; with 
mercuric chloride a white; with picric acid, a yellow, 
which gradually formed in crystalline tablets. This sub- 
stance did not contain any arsenic, but was highly poi- 
sonous. From the stomach of a hog, which had been pre- 
served in a solution of arsenious acid, Selmi separated an 
arsenical organic base. The fluid was distilled in a current 
of hydrogen. The distillate, which was found to be strongly 
alkaline, was neutralized with hydrochloric acid and evapo- 
rated to dryness, when cross-shaped crystals, giving an odor 
similar to that of trimethylamine, were obtained. This sub- 
stance was found by Ciaccia to be highly poisonous, pro- 
ducing strychnia-like symptoms. With iodine in hydriodic 
acid it is said to give a gray, crystalline precipitate. 
From the liquid which remained in the retort, a non- 
volatile arsenical ptomaine was extracted with ether. An 
aqueous solution of this gave with tannic acid a slowly 
forming, yellowish precipitate, and similarly colored pre- 
cipitates with iodine in hydriodic acid, platinum chloride, 
auric chloride, mercuric chloride, potassio-mereurie iodide, 
potassio-bismuthic iodide, picric acid, and potassium bi- 
chromate. The physiological action of this substance as 
demonstrated on frogs was unlike that of the arsines, but 
consisted of torpor and paralysis. 
Moriggia and Battistini experimented with alkaloids, 
obtained from decomposing bodies, upon guinea-pigs and 
frogs, but did not attempt their isolation because of the 
rapid decomposition which they undergo when exposed to 
the air and by which they lose their poisonous properties. 
These alkaloids they found to be easily soluble in amylic 
alcohol, less soluble in ether. 
In 1871 Lombroso showed that the extract from mouldy 
corn-meal produced tetanic convulsions in animals. This HISTORICAL SKETCH. 
33 
threw some light upon the eases of sporadic illness which 
had long been known to occur among the peasants of Lom- 
bardy, who eat fermented and mouldy corn-meal. In 1876 
Brugnatelli and Zenoni obtained by the Stas-Otto 
method from this mouldy meal an alkaloidal substance 
which was white, non-crystalline, unstable, and insoluble 
in water, but readily soluble in alcohol and ether. With 
sulphuric acid and bichromate of potassium it yields a 
color reaction very similar to that of strychnine. 
The action of the ether extracts from decomposed brain 
resembles that of curare, but is less marked and more 
transitory. The beats of the frog’s heart were decreased in 
number and strengthened in force; the nerves aud the 
muscles lost their irritability, and the animal passed into 
a condition of complete torpor. The pupils were dilated. 
Guareschi and Mosso, using the Stas-Otto method, 
obtained from human brains which had been allowed to 
decompose at a temperature of from 10° to 15° for from 
one to two months, both volatile and non-volatile bases. 
Among the former only ammonia and trimethylamine were 
in sufficient quantity for identification. With these, how- 
ever, were minute traces of ptomaines. 
They obtained non-volatile bases from both acid and 
alkaline solutions. From the former they separated a sub- 
stance which gave precipitates with gold chloride, phospho- 
tungstic acid, phospho-molybdic acid, Mayer’s reagent, 
palladium chloride, picric acid, iodine in potassium iodide, 
and slightly with tannic acid. This substance was not 
precipitated with platinum or mercury. 
From the alkaline extract there was obtained a substance 
which in dilute hydrochloric acid solutions gave with gold 
chloride a heavy yellow precipitate with reduction, also 
precipitates with phospho-molybdic acid, platinum chloride, 
Mayer’s reagent, picric acid, phospho-tungstic acid, 
Marme’s reagent, iodine in potassium iodide, tannin, bi- 
chromate of potassium, palladium chloride, and mercuric 
chloride. It reduces ferric salts. From decomposed fibrin 
the same investigators obtained one well-defined ptomaine. 
Analyses of the platinum compound of this substance gave 34 
bacterial poisons. 
the formula C10H15N. This substance will be discussed in 
a future chapter. 
From fresh brain substance they separated ammonia, 
trimethylamine, and an undetermined base. These, how- 
ever, are not to be regarded as products of putrefaction, 
but as resulting from the action of the reagents upon the 
brain substance. The trimethylamine probably arises from 
the splitting up of lecithin, while the undetermined base 
is most likely choline, which also results from the breaking 
up of the lecithin molecule. 
They also show that when Dragendorff’s method is 
used basic substances can be obtained from fresh meat, and 
these are shown to be produced by the action of the sul- 
phuric acid on the flesh. 
To Brieger, of Berlin, is due the credit of isolating 
and determining the composition of a number of ptomaines. 
From putrid flesh he obtained neuridine, C5II14N2, and 
neurine, C5H13NO. The former is inert, while the latter is 
poisonous. From decomposed fish lie separated a poisonous 
base, C2H4(NH2)2, which is an isomerideof ethylenediamine, 
muscarine, C5H15N03, and an inert substance, C7H17N02, 
gadinine. Rotten cheese yielded neuridine and trimethyla- 
mine. Decomposed glue gave neuridine, dimethylamine, 
and a muscarine-like base. In the cadaver, he has found 
in different stages of decomposition, choline, neuridine, tri- 
methylamine, cadaverine, C5H14N2, putrescine, C4H,2K2, and 
saprine, C5H16N2. These are all inert. After fourteen 
days of decomposition he found a poisonous substance, 
mydaleine. From a cadaver which had been kept at 
from —9° to +5° for four months, Brieger obtained 
mydine, C8HnNO, the poisonous substance mydatoxine, 
06K13N02, also the poison methyl-guanidine. From 
poisonous mussel he separated mytilotoxine, C6H15N02. 
From pure cultures of the typhoid bacillus of Koch and 
Eberth, Brieger obtained a poison, typhotoxine, and, 
from like cultures of the tetanus germ of Rosenbach, 
tetanine. All of these bases will be discussed in detail in 
a subsequent chapter. historical sketch. 
35 
Gautier and Etard have also isolated ptomaines which 
will be described later. 
In 1885, Ar a ugh an succeeded in isolating an active 
agent from poisonous cheese, to which he gave the name 
tyrotoxicon. This discovery has been confirmed by New- 
ton, Wallace, Schaffer, Stanton, Firtii, Ladd, 
Wolff, Kl\{ura, Davis, and Kinnicutt. 
Nicati and Kietsch, Koch, and others, have shown 
the presence of a poisonous substance in cultures of the 
cholera bacillus. Salmon and Smith have done the same 
with cultures of the swine-plague germ ; Hoffa, with those 
of the anthrax bacillus; and Brieger with those of the 
tetanus germ. 
In 1888, Christmas obtained from cultures of the 
staphylococcus pyogenes aureus a proto id which, when in- 
jected into the anterior chamber of the eye or under the 
skin, causes suppuration. 
In 1889, Hankin isolated from cultures of the bacillus 
anthraeis a poisonous albumose, which, when employed in 
large doses, proves fatal, and in small doses gives immunity. 
In 1888, Koux and Yersin showed that the chemical 
poison of Loffler’s diphtheria bacillus is a proteid body 
which they believed to be of the nature of a ferment. In 
1890, this work was continued by Brieger and Frankel 
in their memorable contribution on bacterial poisons, in 
which they detail the methods by which they isolate their 
“ toxalbumins” from cultures of the Loffler bacillus, 
the anthrax bacillus, Eberth’s germ, the cholera vibrio, 
and the staphylococcus pyogenes aureus. Martin made a 
more detailed study of the albumoses of anthrax. Vaughan 
reported poisonous proteids in cultures of two toxicogenic 
germs found in drinking-water, also in cultures of three of 
Booker’s summer diarrhoea germs and in poisonous cheese. 
Novy and Schweinitz found both basic and proteid 
poisons in cultures of the swine-plague bacillus. 
Many other contributions have been made, many of 
which will be mentioned in subsequent chapters. CHAPTER III. 
FOODS CONTAINING BACTERIAL POISONS. 
Poisonous Mussels.—Judging from the symptoms 
produced, there seem to be three different kinds of poison- 
ous mussel. In one class, the symptoms resemble those of 
a true gastro-intestinal irritant. Fodere reports the case of 
a sailor, who, after eating a large dish of mussels, suffered 
from nausea, vomiting, pain in the stomach, tenesmus, and 
rapid pulse. After death, which occurred within two days, 
the stomach and intestines were found inflamed and filled 
with a tenacious mucus. Combe and others also report 
cases of the choleraic form of poisoning from mussel. 
However, the symptoms which most frequently manifest 
themselves after the eating of poisonous mussels are more 
purely nervous. A sensation of heat and itching appears 
usually in the eyelids, and soon involves the whole face, 
and perhaps a large portion of the body. An eruption, 
usually called nettle-rash, though it may be papular or 
vesicular, covers the parts. The itching is most annoying, 
and may be accompanied by marked swelling. There 
follows a distressing asthmatic breathing, which is relieved 
by ether. In some cases reported by Mohring, dyspnoea 
])receded the eruption, the patients became insensible, the 
face livid, and convulsive movements of the extremities 
were noticed. Burrow reports similar cases with delirium, 
convulsions, coma, and death within three days. 
In a third class of cases, there may be a kind of intoxi- 
cation resembling somewhat that of alcohol, then paralysis, 
coma, and death. 
In 1827, Combe observed thirty persons poisoned, two 
of them fatally, with mussels. He describes the symptoms 
as follows: “None, so far as I know, complained of any- 
thing peculiar in the smell or taste of the animals, and POISONOUS MUSSELS. 
37 
none suffered immediately after taking them. In general, 
an hour or two elapsed, sometimes more; and the bad 
effects consisted rather in uneasy feelings and debility than 
in any distress referable to the stomach. Some children 
suffered from eating only two or three; and it will be re- 
membered that Robertson, a young and healthy man, only 
took five or six. In two or three hours they complained 
of a slight tension at the stomach. One or two had cardi- 
algia, nausea, and vomiting; but these were not general or 
lasting symptoms. They then complained of a prickly feel- 
ing in their hands, heat and constriction of the mouth and 
throat; difficulty of swallowing and speaking freely; numb- 
ness about the mouth, gradually extending to the arms, with 
great debility of the limbs. The degree of muscular de- 
bility varied a good deal, but was an invariable symptom. 
In some it merely prevented them from walking firmly, 
but in most of them it amounted to perfect inability to 
stand. While in bed they could move their limbs with 
tolerable freedom, but on being raised to the perpendicular 
posture they felt their limbs sink under them. Some com- 
plained of a bad, coppery taste in the mouth, but in general 
this was in answer to what lawyers call a leading question. 
There was slight pain of the abdomen, increased on pres- 
sure, particularly in the region of the bladder, which organ 
suffered variously in its functions. In some the secretion 
of urine was suspended, in others it was free, but passed 
with pain and great effort. The action of the heart was 
feeble; the breathing unaffected; the face pale, expressive 
of much anxiety; the surface rather cold; the mental 
faculties unimpaired. Unluckily, the two fatal cases were 
not seen by any medical person; and we are, therefore, 
unable to state minutely the train of symptoms. We ascer- 
tained that the woman, in whose house were five sufferers, 
went away as in a gentle sleep, and that a few moments 
before death she had spoken and swallowed.” 
The woman died within three hours, aud the other death 
was that of a watchman, who was found dead in his box 
six or seven hours after he had eaten the mussels. Post- 38 
BACTERIAL POISONS. 
mortem examination in these showed no abnormality. The 
stomach contained some of the food partially digested. 
The explorer Vancouver reports four cases similar to 
those observed by Combe. One of the sailors died in five 
and a half hours after eating the mussels. 
In some recent cases reported by Schmidtmann, as 
quoted by B rieger, the symptoms were as follows : Some 
dock hands and their families ate of cooked blue mussels 
which had been taken near a newly built dock. The 
symptoms appeared, according to the amount eaten, from 
soon after eating to several hours later. There was a sen- 
sation of constriction in the throat, mouth, and lips; the 
teeth were set on edge, as though sour apples had been 
eaten. There was dizziness, no headache; a sensation of 
flying, and an intoxication similar to that produced by 
alcohol. The pulse was hard, rapid (eighty to ninety), no 
elevation of temperature, the pupils dilated and reaction- 
less. Speech was difficult, broken, and jerky. The limbs 
felt heavy ; the hands grasped spasmodically at objects and 
missed their aim. The legs were no longer able to support 
the body, and the knees knocked together. There was 
nausea, vomiting, no abdominal pain, no diarrhoea. The 
hands became numb and the feet cold. The sensation of 
cold soon extended over the entire body, and in some the 
perspiration flowed freely. There was a feeling of suffoca- 
tion, then a restful and dreamless sleep. One person died 
in one and three-quarters of an hour, another in three and 
one-half hours, and a third in five hours, after eating of 
the mussels. 
In one of these fatal cases rigor mortis was marked and 
remained for twenty-four hours. The vessels of all the 
organs were distended, only the heart was empty. Vir- 
chow concluded from the conditions observed that the 
blood had absorbed oxygen with great avidity. There was 
marked hypenemia and swelling of the mucous membrane 
of the stomach and intestines, which Virchow pronounced 
an enteritis. The spleen was enormously enlarged and the 
liver showed numerous hemorrhagic infarctions. 
Many theories have been advanced to account for poison- 39 
POISONOUS MUSSELS 
ous mussels. It was formerly believed that the effeets were 
due to copper which the animals obtained from the bottoms 
of vessels; but, as Christison remarks, copper does not 
produce these symptoms. Moreover, Christison made 
analysis of the mussels which produced the symptoms ob- 
served by Combe, and was unable to detect any copper. 
Bouchardat found copper in some poisonous mussels, but 
he does not state the amount of the copper nor the source 
of the animals. 
Edwards advanced the theory that the symptoms were 
wholly due to idiosyncrasy in the consumer. This may be 
true in some instances where only one or two of those par- 
taking of the food are affected, but it certainly is not a 
tenable hypothesis in such instances as those reported by 
Combe aud Schmidtmann, where a large number or all 
those who partook of the food were affected. 
Coldstream found the livers of the Leith mussels, as 
he thought, larger, darker, and more brittle than normal, 
and to this diseased condition he attributed the ill effects. 
Lamoroux, Mohring, de Beume, Chenu, and du 
Rondeau have supposed that the poisonous effects were 
due to a particular species of medusae upon which the mus- 
sels feed. De Beume found in the vomited matter of one 
person, suffering from mussel poisoning, some medusae, and 
he states that these are most abundant during the summer, 
when mussels are most frequently found to be poisonous. 
The theory of Burrow that the animal is always poison- 
ous during the period of reproduction has been received 
with considerable credit. However, cases of poisoning have 
occurred at different seasons of the year. 
Crumpe, in 1872, suggested that there is a species of 
mussel which is in and of itself poisonous, and this species 
is often mixed with the edible variety. Schmidtmann and 
Virchow support this idea. They state that the poisonous 
species has a brighter shell, a sweeter, more penetrating, 
bouillon-like odor than the edible kind, also that the flesh 
of the former is yellow aud that the water in which they 
are cooked is bluish. Loiimeyer also champions this 
opinion. This theory, however, is opposed by the majority 40 
bacterial poisons. 
of zoologists. Mobius states that the peculiarities of the 
supposed poisonous variety pointed out by Virchow and 
Schmidtmann are really due to the conditions under which 
tie animal lives, the amount of salt in the water, the tem- 
perature of the water, whether it is moving or still water, 
the nature of the bottom, etc. Finally, Mobius states that 
the sexual glands, which form the greater part of the 
mantle, are white in the male and yellow in the female. 
However, it has been shown later by Schmidtmann and 
Virchow that edible mussels may become poisonous if left 
in filthy water for fourteen days or longer, and, on the 
other hand, poisonous ones may become fit for food if kept 
for four weeks in good water. 
Cats and dogs which have eaten voluntarily of poisonous 
mussels have suffered from symptoms similar to those ob- 
served in man ; and rabbits have been poisoned by the 
administration of the water in which the food has been 
cooked. A rabbit which was treated in this manner by 
Schmidtmann died within one minute. From these 
mussels Brieger extracted the ptomaine mytilotoxine, 
which will be discussed in a subsequent chapter. This 
poison has a curare-like action. Whether or not those 
mussels which produce other symptoms also contain pto- 
maines, remains for future investigations to determine. 
In 1887 three other cases of mussel poisoning, one fatal 
case, occurred at Wilhelmshaven, the place which supplied 
Brieger with the mussels from which he obtained mytilo- 
toxine. Schmidtmann has found that non-poisonous 
mussels placed in the waters of this bay soon become poi- 
sonous, and that the poisonous mussels from the bay placed 
in the open sea soon lose their poisonous properties. Lin- 
der has found in the water of the bay and in the mussels 
living in it a great variety of protozoa, amoeba, bacteria, 
and other lower organisms, which are not found in the 
water of the open sea nor in the non-poisonous mussel. He 
has also found that, if the water of the bay be filtered, non- 
poisonous mussels in it do not become poisonous. He 
therefore concludes that poisonous mussels are those which 
are suffering from disease due to residence in filthy water. 41 
POISONOUS FISH. 
Brieger has tested dead and decomposed mussels taken 
from the open sea for mytilotoxine, with negative results. 
Poisonous Oysters and Eels.—Pasquier reported 
cases of poisoning at Havre from the eating of oysters 
taken from an artificial bed which had been established 
near the outlet of a drain from a public water-closet. 
Ciiristison says that an “ unusual prevalence of colic, 
diarrhoea, and cholera” at Dunkirk was believed to have 
been traced to an importation of unwholesome oysters from 
the Normandy coast. Vaughan and Novy obtained 
tests for tyrotoxicon in the liquor of some decomposed 
oysters which had caused illness in mauy people at a church 
festival. 
A7irey states that many persons were attacked with 
violent pain and diarrhoea a few hours after eating a pate 
made of eels from a stagnant cattle-ditch near Orleans, 
also that similar cases have occurred in various parts of 
France, and that domestic animals have been killed by 
eating the remains of the poisonous dish. 
Poisonous Fish.—While many species of fish are popu- 
larly regarded as poisonous, but little scientific work has 
been done iu this line, and we are not prepared to say to 
what extent this popular idea is correct. Mtura and 
Takesaki find that the ripe ovaries of tetrodon rubripes 
contain a substance which induces in rabbits acceleration 
of the respiratory movements, paralysis of the skeletal 
muscles, mydriasis, increased peristalsis of the intestines, 
and arrest of the heart. 
The disease known as “ kakke,” which prevails from 
May to October in Tokio is, according to Miura and 
others, an intoxication due to the eating of fish, which be- 
long to the scombridce. The affection is generally chronic 
or subacute, seldom acute. The most characteristic symp- 
tom is paralysis of the diaphragm with consequent dysp- 
noea and disturbance of the action of the heart. Electri- 
cal stimulation of the diaphragm has proven to be the 
most successful treatment. 42 
BACTERIAL BOISONS. 
Sausage Poisoning.—This is also known as botulis- 
mus and allantiasis. While considerable diversity has 
been observed in symptoms of sausage poisoning, we can- 
not divide the cases into classes from their symptoma- 
tology as has been done in mussel poisoning. The first 
effects may manifest themselves at any time from one hour 
to twenty-four hours after eating of the sausage, and cases 
are recorded in which it is stated, no symptoms appeared 
until several days had passed. However, we must re- 
member that trichinosis was frequently, in former times, 
classed as sausage poisoning, and it is highly probable that 
these cases of long delay in the appearance of the symp- 
toms were really not due to putrefaction, but to the pres- 
ence of parasites in the meat. A large majority of the one 
hundred and twenty-four cases more recently reported by 
Mueger sickened within twenty-four hours, and out of 
the forty-eight of these which were fatal, six died within 
the first twenty-four hours. At first there is dryness of 
the mouth, constriction of the throat, uneasiness in the 
stomach, nausea, vomiting, vertigo, indistinctness of vision, 
dilatation of the pupils, difficulty in swallowing, and 
usually diarrhoea, though obstinate constipation may exist 
from tine first. There is, as a rule, a sensation of suffoca- 
tion, and the breathing becomes labored. The pulse is 
small, thready, and rapid. In some cases the radial pulse 
may be imperceptible. Marked nervous prostration and 
muscular debility follow. These symptoms vary greatly 
in prominence in individual cases. The rechting and vom- 
iting, which may be most distressing and persistent in 
some instances, in others are trivial at the beginning and 
soon cease altogether. The same is true of the diarrhoea. 
As a rule, the functions of the brain proceed normally, but 
there may be delirium, then coma and death. In some 
there are marked convulsive movements, especially of the 
limbs, in others paralysis may be an early and marked 
symptom. The pupils may dilate, then become normal 
and again dilate. There is frequently ptosis, and paralysis 
of the muscles of accommodation is not rare. Complete 
blindness has followed in a few instances. SAUSAGE POISONING. 
43 
The fatality varies greatly in different outbreaks. In 
1820 Kerner collected reports of seventy-six cases, of 
which thirty-seven were fatal. In his next publication 
(1822) he increased the number to one hundred and fifty- 
five cases, with eighty-four fatal results. This gave a 
mortality of over fifty per cent., while in one outbreak 
reported by Muller the mortality was less than two per 
cent. 
A large proportion of the eases of sausage poisoning 
have occurred in Wurtemberg and the immediately adja- 
cent portions of Baden. This fact has, without doubt, 
been correctly ascribed to the methods there practised of 
preparing and curing the sausage. It is said to be com- 
mon for the people to use the blood of the sheep, ox, and 
goat in the preparation of this article of diet. Moreover, 
the blood is kept sometimes for days in wooden boxes and 
at a high temperature before it is used. In these cases it 
is altogether likely that putrefaction progresses to the poi- 
sonous stage before the process of curing is begun. How- 
ever, cases of poisoning have occurred from beef and pork 
sausages as well. 
Moreover, the method of curing employed in Wurtem- 
berg favors putrefaction. A kind of sausage known as 
“blunzen” is made by filling the stomachs of hogs with 
the meat. In curing, the interior of this great mass is not 
acted upon, and putrefaction sets in. The curing is usually 
done by hangiug the sausage in the chimney. At night 
the fire often goes out and the meat freezes. The alternate 
freezing and thawing render decomposition more easy. 
The interior of the sausage is generally the most poison- 
ous. Indeed, in many instances those who have eaten of 
the outer portion have been uuharmed, while those who 
have eaten of the interior of the same sausage have been 
most seriously affected. 
Many German writers state that when a poisonous saus- 
age is cut, the putrid portion has a dirty, grayish-green 
color, and a soft, smeary consistency. A disagreeable 
odor, resembling that of putrid cheese, is perceptible. The 
taste is unpleasant, and sometimes a smarting of the mouth 44 
BACTERIAL POISONS. 
and throat is produced. Post-mortem examination after 
sausage poisoning shows no characteristic lesion. It is 
generally stated that putrefaction sets in very tardily, but 
Muller shows that no reliance can be placed upon this 
point, and states that out of forty-eight recorded autopsies, 
it was especially stated in eleven that putrefaction rapidly 
developed. In some instances there has been noticed 
hypenemia of the stomach and intestinal canal, but this is 
by no means constant. The liver and brain have been re- 
ported as congested, but this would result from the failure 
of the heart, and would, by no means, be characteristic of 
poisoning with sausage. 
Von Faber, in 1821, observed sixteen persons who 
were made sick by eating fresh, uusmoked sausage made 
from the flesh of a pig which had suffered from an abscess 
on the neck. Five of the patients died. The symptoms 
were as follows: There was constriction of the throat, 
difficulty in swallowing, retching, vomiting, colic-like 
pains, vertigo, hoarseness, dimness of vision, and headache. 
Later and in severer cases, there was complete exhaustion, 
and, finally, paralysis. The eyeballs were retracted, the 
pupils were sometimes dilated, then contracted; they did 
not respond to light; there was paralysis of the upper lids. 
The tonsils were swollen, but not as in tonsillitis. Liquids 
which were not irritating could be carried as far as the 
oesophagus, when they were then ejected from the mouth 
and nose with coughing. Solid foods could not be swal- 
lowed. On the back of the tongue and in the pharynx 
there was observed a puriform exudate. 
Obstinate constipation existed in all, while the sphincter 
ani was paralyzed. The breathing was easy, but all had 
a croupous cough. The skin was dry. There was incon- 
tinence of urine. There was no delirium and the mind 
remained clear to the last. 
Post-mortem examinations were held on four. The 
skin was rough—“goose-skin.” The abdomen was re- 
tracted. The large vessels in the upper part of the stom- 
ach were filled with black blood. The contents of the 
stomach consisted of a reddish-brown, semi-fluid substance, SAUSAGE POISONING. 
45 
which gave off a repugnant, acid odor. In one case the 
omentum was found greatly congested. The large intes- 
tine was very pale, and the right ventricle of the heart 
was filled with dark fluid blood. 
Schuz cites thirteen cases of poisoning from liver saus- 
age in which the symptoms differed from the foregoing in 
the following respects : 
(1) In only one out of the thirteen was there constipa- 
tion ; all the others had numerous watery, typhoid-like 
stools. 
(2) Symptoms involving the sense of sight were present 
in only three; in all the pupils were unchanged. 
(3) The croupous cough was wholly wanting; though 
in many there was complete loss of voice. Difficulty of 
swallowing was complained of by only one. 
(4) Delirium was marked in all; and in one the dis- 
turbance of the mental faculties was prominent for several 
weeks. 
(5) There were no deaths. 
(6) The time between eating the sausage and the appear- 
ance of the symptoms varied from eighteen to twenty-four 
hours, and the duration of sickness from one to four 
weeks ; though in one case complete recovery did not occur 
until after two and one-half months. 
The sausages were not smoked, and all observed a garlic 
odor, though no garlic had been added to the meat. 
Tripe reports sixty-four cases. The symptoms came 
on from three and one-half to thirty-six hours after eating. 
The stools were frequent, watery, and of offensive odor. 
In some there was delirium. One died. In the fatal case 
the hands and face were cold and swollen. The pulse was 
rapid and weak. The pupils were contracted, but re- 
sponded to light. The small intestine was found inflamed. 
Hedinger reports the case of a man and a woman with 
the usual symptoms, but during recovery the dilatation of 
the pupils was followed by contraction. Birds ate of this 
sausage, and were not affected. 
Koser reports cases in which there were found, after 
death, abscesses of the tonsils, a dark, bluish appearance 46 
bacterial poisons. 
of the mucous membrane of the pharynx, larynx, and 
bronchial tubes, dark redness of the fundus of the stom- 
ach, and circumscribed, gray, red, and black spots on the 
mucous membrane of the intestine. The liver was brittle 
and the spleen enlarged. 
Many theories concerning the nature of the active prin- 
ciple of poisonous sausage have been advanced. It was 
once believed to consist of pyroligneous acid, which was 
supposed to be absorbed by the meat from the smoke used 
in curing it; but it was soon found that unsmoked sausage 
might be poisonous also. Emmert believed that the active 
agent was hydrocyanic acid, and Jager’s theory supposed 
the presence of picric acid. But these acids are not found 
in poisonous sausage, and, moreover, their toxicological 
effects are wholly unlike those observed in sausage poison- 
ing. As we have elsewhere seen, Kerner believed that 
he had found the poisonous principle in a fatty acid. This 
theory was supported by Dann, Buchner, and Schu- 
mann. Kerner believed the poison to consist of either 
caseic or sebacic acid, or both, while Buchner named it 
acidum botulinicum ; but the acids of the former proved 
to be inert, and that of the latter to have no existence. 
Schlossberger first suggested that the poisonous sub- 
stance is most probably basic in character, and he found 
an odoriferous, ammoniacal base which could not be found 
in good sausage, and which did not correspond to any 
known amides, imides, or nitril bases. However, this 
substance has not been obtained by anyone else, nor has 
it been demonstrated to be poisonous. 
Liebig, Duflas, Hirsch, and Simon believed in the 
presence of a poisonous ferment. N an den Corput de- 
scribed sarcina botulina, which was believed to constitute 
the active agent. Muller, Hoppe-Seyler, and others 
have found various microorganisms, and Virchow, Eich- 
enberg, and others have examined microscopically the 
blood of persons poisoned with sausage. Recently, Ehr- 
enberg has attempted to isolate the poisonous substance 
by employing Brieger’s method, but he obtained only 
inert substances. POISONOUS HAM. 
47 
Gaffky and Paak have made a thorough study of 
some sausage which poisoned a large number of people, 
among whom one, a strong man, died. The sausage was 
made of horse-flesh and liver. In the majority of the 
persons the symptoms came on within six hours and in 
one instance within half an hour. Many had a severe 
chill; some did not. The most prominent symptoms were 
headache, loss of appetite, pain in the bowels, vomiting 
and purging. In the fatal case, however, there was no 
vomiting. From the sausage Gaffky and Paak isolated 
a short bacillus, which when given by the mouth, sub- 
cutaneously or intravenously produced the above symptoms, 
with a fatal termination in most instances, in rabbits, 
guinea-pigs, mice, and apes. Gaffky and Paak were 
unable to isolate the chemical poison. 
Poisonous IIam.—Under this head we shall not discuss 
cases of poisoning from trichina or other parasites, but shall 
refer only to those instances in which the toxic agent has 
originated in putrefactive changes. A number of such 
cases have been observed within the past ten years, but 
only a few of them have been investigated scientifically. 
The best known of these, as well as the most thoroughly 
studied, is the Wellbeck poisoning, which Ballard in- 
vestigated successfully. In June, 1880, a large number of 
persons attended a sale of timber and machinery on the 
estate of the Duke of Portland at Wellbeck. The sale 
continued four days, and lunches were served by the pro- 
prietress of a neighboring hotel. The refreshments con- 
sisted of cold boiled ham, cold, boiled, or roasted beef, 
cold beefsteak pie, mustard and salt, bread and cheese, 
pickles and Chutney sauce. The drinks were bottle and 
draught beer, spirits, ginger beer, lemonade, and water. 
Many were poisoned, and Ballard obtained the particu- 
lars of seventy-two cases, among which there were four 
deaths. The symptoms are given by Ballard as follows: 
“ I propose to speak of the attacks under the name of 
‘diarrhoeal illness/ because diarrhoea was the most constant 
of all the symptoms observed, and the other symptoms 48 
BACTERIAL POISONS. 
were in some respects so peculiar that I am indisposed to 
give to the disease any name otherwise generally recognized. 
As might have been anticipated from our experience of 
diseases in general, there were varieties in severity among 
the cases investigated; and symptoms strongly marked in 
some, were slightly marked or altogether wanting in others. 
Perhaps I shall do the best service by giving first a general 
sketch of the course of the illness, subsequently illustrating 
it by a description of a few well-marked cases. 
“A period of incubation preceded the illness. In fifty- 
one cases where this could be accurately determined, it 
was twelve hours or less in five cases; between twelve and 
thirty-six hours in thirty-four cases; between thirty-six 
and forty-eight hours in eight cases; and later than this in 
only four eases. In many cases the first definite symptoms 
occurred suddenly, and evidently unexpectedly, but in some 
cases there were observed during the incubation more or 
less feeling of languor and ill health, loss of appetite, 
nausea, or fugitive, griping pains in the belly. In about 
a third of the cases the first definite symptom was a sense 
of chilliness, usually with rigors, of trembling, in one case 
accompanied by dyspnoea; in a few cases it was giddiness 
with faintness, sometimes accompanied by a cold sweat and 
tottering; in others, the first symptom was headache or 
pain somewhere in the trunk of the body, e. g., in the 
chest, back, between the shoulders, or in the abdomen, to 
which part the pain, wherever it might have commenced, 
subsequently extended. In one case the first symptom 
noticed was a difficulty in swallowing. In two cases it was 
intense thirst. But however the attack may have com- 
menced, it was usually not long before pain in the abdomen, 
diarrhoea, and vomiting came on, diarrhoea being of more 
certain occurrence than vomiting. The pain in several 
cases commenced in the chest or between the shoulders, and 
extended first to the upper and then to the lower part of 
the abdomen. It was usually very severe indeed, quickly 
producing prostration or faintness, with cold sweats. It 
was variously described as crampy, burning, tearing, etc. 
The diarrhoeal discharges were in some cases quite unre- POISONOUS HAM. 
49 
strainable, and (where a description of them could be ob- 
tained) were said to have been exceedingly offensive and 
usually of a dark color. Muscular weakness was an early 
and very remarkable symptom in nearly all the cases, and 
in many it was so great that the patient could only stand 
by holding on to something. Headache, sometimes severe, 
was a common and early symptom ; and in most cases there 
was thirst, often intense and most distressing. The tongue, 
when observed, was described usually as thickly coated 
with a brown, velvety fur, but red at the tip and edges. 
In the early stage the skin was often cold to the touch, but 
afterward fever set in, the temperature rising in some 
cases to 101°, 103°, aud 104° F. In a few severe cases 
where the skin was actually cold, the patient complained of 
heat, insisted on throwing off the bedclothes, and was very 
restless. The pulse in the height of the illness became 
quick, counting in some cases 100 to 128. The above 
were the symptoms most frequently noted. Other symp- 
toms occurred, however, some in a few cases, and some only 
in solitary cases. These I now proceed to enumerate. 
Excessive sweating, cramps in the legs, or in both legs and 
arms, convulsive flexion of the hands or fingers, muscular 
twitchiugs of the face, shoulders, or hands, aching pain in 
the shoulders, joints, or extremities, a sense of stiffness of 
the joints, prickling or tingling or numbness of the hands 
lasting far into convalescence in some cases, a sense of 
general compression of the skin, drowsiness, hallucinations, 
imperfection of vision, and intolerance of light. In three 
cases (one, that of a medical man) there was observed yel- 
lowness of the skin, either general or confined to the face 
aud eyes. In one case, at a late stage of the illness, there 
was some pulmonary congestion, and an attack of what was 
regarded as gout. Iu the fatal cases, death was preceded 
by collapse like that of cholera, coldness of the surface, 
pinched, features and blueness of the fingers and toes and 
around the sunken eyes. The debility of convalescence 
was in nearly all cases protracted to several weeks. 
“The mildest cases were characterized usually by little 
remarkable beyond the following symptoms, viz., abdominal 50 
bacterial poisons. 
pains, vomiting, diarrhoea, thirst, headache, and muscular 
weakness; any one or two of which might be absent.” 
The cause of this illness was traced conclusively to the 
hams eaten. Klein found in the meat a bacillus, cultures 
of which were used for inoculating animals. These inocu- 
lations were found generally to be followed by pneumonia. 
No attempt was made to isolate a ptomaine. 
Later, Ballard reported fifteen cases with symptoms 
similar to the above, and with one death, from eating baked 
pork. Not all of those who ate of this pork were made 
sick. This might have been due to inequality in the putre- 
factive changes in different portions of the meat, or it may 
have been due to differences in temperature in various por- 
tions of the meat during the cooking. In the blood, peri- 
cardial fluid, and lungs of the fatal ease, Klein observed 
bacilli similar to those discovered in the Wellbeck inquiry. 
Pneumonia was produced by inoculating guinea-pigs and 
mice with these bacilli. 
In meat which poisoned a large number of persons, 
Gartner found his bacillus euteritidis. The meat was 
from a cow which had a severe diarrhoea for two days be- 
fore she was killed. Of twelve persons who ate the flesh 
raw, all were sick; while of those who ate of the cooked 
food a large per cent, were also affected. In the meat and 
in the spleen of a person who died from the effects of the 
poison, Gartner found the bacillus, which proved fatal 
to animals. Good beef, inoculated with this bacillus and 
cooked some hours later, killed rabbits, guinea-pigs, and 
mice. The skin of the people who were poisoned and re- 
covered peeled off. The period of incubation varied from 
two to thirty hours. 
August 29, 1887, 256 soldiers and 36 citizens at Middle- 
burg, Holland, were taken sick after eating meat from a 
cow which had been killed while suffering from puerperal 
fever. The symptoms were nausea, vomiting, purging, 
elevation of temperature, and prostration. In some there 
were observed dizziness, sleepiness, and dilatation of the 
pupil. After a few days these symptoms gradually disap- 
peared, and in many an eczematous eruption of the lips 51 
POISONOUS MEAT. 
gave annoyance. Pigs, cats, and dogs which ate of the 
offal of tliis animal were also made sick. Thorough 
cooking did not destroy the poison, and those who took 
soup and boullion made from the meat were affected like 
those who ate of the muscular fibre. In most of the cases 
the symptoms came on within twelve hours after eating 
the meat. 
On a fete-day at Zurich, in 1839, 600 persons who were 
fed upon cold veal and ham were taken ill, with shivering, 
giddiness, vomiting, and diarrhoea. Some were delirious 
and others were salivated, the saliva being extremely fetid. 
In the worst cases there were involuntary stools, collapse, 
and death. The cause was traced to putrefactive changes 
in the meat. 
SiedIjER reports an instance of four persons having been 
made sick by eating decomposed goose-grease. There were 
giddiness, prostration, and violent vomiting. No metallic 
poison could be found. The grease was rancid, of repul- 
sive odor, and three ounces of it given to a dog produced 
the same symptoms which had been observed in the 
persons. 
Christison reports a number of cases in which persons 
were seriously, a few fatally, affected by eating various 
kinds of meat which had undergone partial putrefaction. 
Ollivier found six persons poisoned, four of them 
fatally, by eating of decomposed mutton. He also men- 
tions the poisoning of a family of three with ham pie. 
Chemical analysis failed to reveal the presence of any 
poison. 
Boutigny, having failed to find any poison in the meat 
furnished at a festival, and to which the serious illness of 
many was attributed, made a meal of stuffed turkey fur- 
nished by the same dealer, but after a short time his coun- 
tenance became livid, his pulse small aud feeble, a cold 
sweat bathed his body, aud violent vomiting and purging 
followed. His recovery was slow. 
Gjrserer observed nausea, vomiting, purging, and 
delirium after eating of bacon which was imperfectly cured. 52 
BACTERIAL POISONS. 
Poisonous Canned Meats.—Cases of poisoning from 
eating canned meats have become quite frequent. Although 
it may be possible that in some instances the untoward 
effects result from metallic poisoning, in the great majority 
of cases the poisonous principles are formed by putrefactive 
changes. In many instances it is probable that decomposi- 
tion begins after the can is opened by the consumer. In 
others, the canning is carelessly done and putrefaction is 
far advanced before the food reaches the consumer. In 
still other instances, the meat may be taken from diseased 
animals, or it may undergo putrefactive changes before 
the canning. What is true of canned meats is also true of 
canned fruits and vegetables. 
Dr. Ashworth, of Smithland, Iowa, has reported to us 
three fatal cases of poisoning from canned apricots. An 
infant, which was only eight days old, and which must 
have received the poison from its mother’s breasts, died 
within a few hours. The mother died forty-three hours 
after eating the apricots, and the father on the sixth day. 
The symptoms corresponded with those of poisoning by 
tyrotoxicon. However, it seems that no analysis was 
made, and these may have been cases of mineral poisoning. 
Poisonous Cheese.—In 1827 Hunnefeld made some 
analyses of poisonous cheese, and experimented with ex- 
tracts upon the lower animals. He accepted the ideas of 
Kerner in regard to poisonous sausage in a somewhat 
modified form, and thought the active agents to be sebacic 
and caseic acids. About the same time, Serturner, 
making analyses of poisonous cheese for Westrumb, also 
traced the poisonous principles, as he supposed, to these 
fatty acids. We see from this that during the first part of 
the present century the fatty acid theory, as it may be 
called, was generally accepted. 
In 1848, Christison, after referring to the work of 
Hunnefeld and Serturner, made the following state- 
ment : “His (Hiinnefeld’s) experiments, however, are not 
quite conclusive of the fact that these fatty acids are really 
the poisonous principles, as he has not extended his experi- poisonous cheese 
53 
mental researches to the caseie and sebacic acids prepared 
in the ordinary way. His views will probably be altered 
and simplified if future experiments should confirm the 
late inquiries of Braoonnot, who has stated that Proust’s 
caseic acid is a modification of acetic acid combined with 
an acrid oil.” 
In 1852 Schlossberoer made experiments with the 
pure fatty acids and demonstrated their freedom from poi- 
sonous properties. These experiments have been verified 
repeatedly, so that now it is well known that all the fatty 
acids obtainable from cheese are devoid of poisonous 
properties. 
It may be remarked here, that there is every probability 
that the poisonous substance was present in the extracts 
obtained by the older chemists. Indeed, we may say that 
this is a certainty, since the administration of these extracts 
to cats was, in some instances at least, followed by fatal 
result. The great mass of these extracts consisted of fatty 
acids, and as the chemists could find nothing else present, 
they very naturally concluded that the fatty acids them- 
selves constituted the poisonous substance. 
Since the overthrow of the fatty acid theory, various 
conjectures have been made, but none worthy of considera- 
tion. 
We make the following quotations from some of the best 
authorities who wrote during the first half of the past 
decade upon this subject: 
IlrLLER says : “ Nothing definite is known of the nature 
of cheese poison. Its solubility seems established from an 
observation by ITusemann, a case in which the poison was 
transmitted from a nursing mother to her child.” 
Husemann wrote as follows : u The older investigations 
of the chemical nature of cheese poison, which led to the 
belief of putrefactive cheese acids and other problematic 
substances, are void of all trustworthiness, and the dis- 
covery of the active principle of poisonous cheese may not 
be looked for in the near future, on account of the proper 
animals for controlling the experiments with the extracts, 54 
BACTERIAL POISONS. 
as dogs can eat large quantities of poisonous cheese without 
its producing any effect.” 
Brieger stated in 1885: “All kinds of conjectures con- 
cerning the nature of this poison have been formed, but all 
are even devoid of historical interest; because they are not 
based upon experimental investigations. My own experi- 
ments toward solving this question have not progressed 
very far.” 
In the above quotation we think that Brieger has 
hardly done justice to the work of IIcnnefeld and Ser- 
turneii. Their labors can hardly be said to be wholly 
devoid of historical interest, and they certainly did employ 
the experimental method of inquiry. 
In the years 1888 and 1884 there were reported to the 
Michigan State Board of Health about three hundred cases 
of cheese poisoning. As a rule, the first symptoms ap- 
peared within from two to four hours after eating the 
cheese. In a few the symptoms were delayed from eight 
to ten hours and were very slight. The attending physi- 
cians reported that the gravity of the symptoms varied with 
the amount of cheese eaten, but no one who ate of the 
poisonous cheese wholly escaped. One physician reported 
the following symptoms: “Everyone who ate of the 
cheese was taken with vomiting, at first of a thin, watery, 
later a more consistent reddish-colored substance. At the 
same time the patient suffered from diarrhoea with watery 
stools. Some complained of pain in the region of the 
stomach. At first the tongue was white, but later it be- 
came red and dry, the pulse was feeble and irregular; 
countenance pale, with marked cyanosis. One small boy, 
whose condition seemed very critical, was covered all over 
the body with bluish spots.” 
Dryness and constriction of the throat were complained 
of by all. In a few cases the vomiting and diarrhoea were 
followed by marked nervous prostration, and in some dila- 
tation of the pupils was observed. 
Notwithstanding the severity of the symptoms in many, 
there was no fatal termination among these cases, though 
several deaths from cheese poisoning in other outbreaks POISONOUS CHEESE. 
55 
have occurred. Many of the physicians at first diagnosed 
the cases from the symptoms as due to arsenical poisoning, 
and on this supposition some admiuisteted ferric hydrate. 
Others gave alcohol and other stimulants and treated upon 
the expectant plan. 
Vaughan, to whom the cheese was sent for analysis, 
made the following report: “All of these three hundred 
cases were caused by eating of twelve different cheeses. Of 
these, nine were made at one factory, and one each at three 
other factories. Of each of the twelve I received smaller 
or larger pieces. Of each of ten I received only small 
amounts. Of each of the other two I received about 
eighteen kilogrammes. The cheese was in good condition 
and there was nothing in the taste or odor to excite sus- 
picion. However, from a freshly cut surface there exuded 
numerous drops of a slightly opalescent fluid which red- 
dened litmus paper instantly and intensely. Although, as 
I have stated, I could discern nothing peculiar in the odor, 
if two samples, one of good, the other of poisonous cheese, 
were placed before a dog or cat, the animal would invari- 
ably select the good cheese. But if only poisonous cheese 
was offered, and the animal was hungry, it would partake 
freely. A cat was kept seven days and furnished only 
poisonous cheese and water. It ate freely of the cheese 
and manifested no untoward symptoms. After the seven 
days the animal was etherized and abdominal section was 
made. Nothing abnormal could be found. I predicted, 
however, in one of my first articles on poisonous cheese, 
that the isolated poison would affect the lower animals. 
As to the truth of this prediction we will see later. 
“ My friend, Dr. Sternberg, the eminent bacteriologist, 
found in the opalescent drops above referred to numerous 
micrococci. But inoculations of rabbits with these failed 
to produce any results. 
“At first I made an alcoholic extract of the cheese. After 
the alcohol was evaporated in vacuo at a low temperature 
a residue consisting mainly of fatty acids remained. I ate 
a small bit of this residue, and found that it produced dry- 
ness of the throat, nausea, vomiting, and diarrhoea. The 56 
BACTERIAL POISONS. 
mass of this extract consisted of fats and fatty acids, and 
for some weeks I endeavored to extract the poison from 
these fats, but all attempts were unsuccessful. I then made 
an aqueous extract of the cheese, filtered this, and drinking 
some of it, found that it also was poisonous. But after 
evaporating the aqueous extract to dryness on the water- 
bath at 100°, the residue thus obtained was not poisonous. 
From this I ascertained that the poison was decomposed or 
volatilized at or below the boiling-point of water. I then 
tried distillation at a low temperature, but by this the 
poison seemed to be decomposed. 
“ Finally, I made the clear, filtered aqueous extract, 
which was highly acid, alkaline with sodium hydrate, agi- 
tated this with ether, removed the ether, and allowed it to 
evaporate spontaneously. The residue was highly poison- 
ous. By re-solution in water and extraction with ether, the 
poison was separated from foreign substances. As the ether 
took up some water, this residue consisted of an aqueous 
solution of the poison. After this was allowed to stand for 
some hours in vacuo over sulphuric acid, the poison sepa- 
rated in needle-shaped crystals. From some samples the 
poisoned crystallized from the first evaporation of the ether, 
and without standing in vacuo. This happened only when 
the cheese contained a comparatively large amount of the 
poison. Ordinarily, the microscope was necessary to detect 
the crystalline shape. From sixteen kilogrammes of one 
cheese, I obtained about 0.5 gramme of the poison, and in 
this case the individual crystals were plainly visible to the 
unaided eye. From the same amount of another cheese I 
obtained only about 0.1 gramme, and the crystals in this 
case were not so large. 1 have no idea, however, that by 
the method used all the poison was separated from the 
cheese.” 
To this ptomaine Vaughan has given the name tyro- 
toxicon (Tvpog, cheese, and poison). Its chemistry 
will be discussed in a subsequent chapter. 
During 1887, Wallace found tyrotoxicon in two 
samples of cheese which had caused serious illness. The 
first of these came from Jeauesville, Pa., and the symptoms POISONOUS CHEESE. 
57 
as reported to Wallace by Doolittle, who had charge 
of the cases, were as follows: “ There were at least fifty 
persons poisoned by this cheese. There were also eight 
others who ate of the cheese, but felt no unpleasant effects; 
whether this was due to personal idiosyncrasy, or to an 
uneven distribution of the poison throughout the cheese, I 
am unable to say. 
“ The majority, however, comprising fifty or sixty per- 
sons, were seized, in from two to four hours after eating the 
cheese, with vertigo, nausea, vomiting, and severe rigors, 
though varying in their order of appearance and in severity 
in different cases. The vomiting and chills were the most 
constant and severe symptoms in all the eases, and were 
soon followed by severe pain in the epigastric region, 
cramps in the feet and lower limbs, purging and griping 
pain in the bowels, a sensation of numbness or pins and 
needles, especially in the limbs, and lastly, very marked 
prostration, amounting almost to collapse in a few cases. 
“ The vomit at first consisted of the contents of the 
stomach, and had a strong odor of cheese; afterward it 
consisted of mucus, bile, and in three or four of the severer 
cases blood was mixed with the mucus in small quantities. 
Microscopic examination of the same was not made, but to 
the eye it appeared as such. The vomiting and diarrhoea 
lasted from two to twelve hours; the rigors and muscular 
cramps, one to two hours. The diarrhceal discharges, at 
first fecal, became later watery and light colored. No 
deaths occurred, and for the most part the effects were 
transient, and all that remained on the following day were 
the prostration and numbness; the latter occurred in about 
one-half the cases, and disappeared in from one to three 
days. 
“Children, as a rule, seemed to suffer less than adults, 
and, of course, it was not possible to elicit as definite symp- 
toms from them. The suddenness of the attack was 
remarked by all, some feeling perfectly well until the 
moment of attack. Nor did the symptoms seem to be in 
proportion to the amount of cheese taken ; some of the 
severest cases declared they had not eaten more than a cubic 58 
BACTERIAL POISONS. 
inch of it. One of the severest cases was about six and 
one-half months pregnant, but no interference with preg- 
nancy occurred. All the cheese which caused the sickness 
came from the same piece.” 
The second sample of cheese examined by Wallace 
came from Riverton, N. J. This outbreak included a 
smaller number of persons, all of whom recovered. 
Wolff has detected tyrotoxicon in cheese which poisoned 
several persons at Shamokin, Pa. The pores of this cheese 
were found filled with a grayish-green fungoid growth, 
though it is not supposed that this fungus was connected in 
any way with the poisonous nature of the cheese. Tests 
were made for mineral poison with negative results, after 
which tyrotoxicon was recognized both by chemical and 
physiological tests. “ A few drops of the liquid (extract), 
placed on the tongue of a young kitten, produced prompt 
emesis and numerous watery dejections with evident depres- 
sion and malaise of the animal. A larger cat was similarly 
affected by it, though the depression and malaise were not 
so marked nor so long continued.” 
Cheese poisoning caused the death of several children in 
the neighborhood of Heiligenstadt, in 1879, and there were 
many fatal cases from the same cause in Pyrmont, in 1878. 
Unfortunately we have not been able to find any detailed 
account of either the symptoms or the post-mortem appear- 
ances in these cases. 
Ehrhart has published the history of some cases of 
poisoning from cheese, of which the following is an abstract: 
The family of a workman, consisting of eight persons, ate for 
supper 600 grammes (about eighteen ounces) of Limburger 
cheese. The rind was covered with a heavy mould, while 
the interior had become fluid from putrefaction, and was of 
bitter taste. Three ate only of the mouldy rind, and these 
remained well. The next morning, the five who had eaten 
of the inner portion suffered from vertigo, nausea, vomiting, 
and abdominal pains; no stool. The father had convulsive 
movements of all the extremities. The pupils were dilated, 
and did not respond to light; there were double vision, 
cold sweat, skin cyanotic, abdomen distended, difficulty in POISONOUS CHEESE. 
59 
swallowing, delirium, mild trismus, and temperature 40° C. 
(104° F.). The temperature of the mother, on account of 
the great collapse, was subnormal. She had no convulsive 
movements, but there was prolonged loss of consciousness. 
The pulse was small and thready, and threatened paralysis 
of the heart. Recovery was very slow. The others suf- 
fered only from gastro-enteric symptoms. Ehrhardt 
discusses the question as to whether these symptoms were 
due to tyrotoxicon, or to infection with microorganisms; 
but as we have not had access to his original paper, we do 
not know what his conclusions are. However, there cannot 
be much doubt that in those cases in which the organism 
is taken into the alimentary canal, it continues the elabora- 
tion of its poisonous products. 
In 1890 Vaughan made the following additional report 
on poisonous cheese: 
‘‘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. The evidence 
that these samples had been the actual cause of the sickness 
among the people who had eaten of them was thus con- 
firmed by the experiments 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 demon- 
strate the nature of the poison. Tyrotoxicon could not be 
detected.’ 
“ One sample of this class was found by Hovy 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 grammes of the 
cheese was 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°. 60 
BACTERIAL POISONS. 
for twenty-four hours, became so poisonous that 100 c. c. of 
it introduced into the stomach of a full-grown cat caused 
death. Novy made plate cultures from the cheese and 
from the spleen and liver of the dead animals, and suc- 
ceeded in identifying one germ as common to both. Ster- 
ilized 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 prop- 
erties, 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 appreciable effect. The report made upon the 
samples was as follows : 
“ ‘ Animals fed upon the cheese were not affected. Tyro- 
toxicon could not be found. The sickness in 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 kilo- 
grammes of a cheese of this class was 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 super- 
natant fluid was decanted, and the precipitate was dissolved 
in water and re-precipitated 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 POISONOUS CHEESE. 
61 
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 alter some 
days seemed to be wholly recovered. 
“ This substance belongs to the so-called poisonous albu- 
mins. 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 satura- 
tion 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 this manner get enough of the poison to affect 
them. It cannot be said positively that the samples of 
cheese of the first class mentioned above owe their poison- 
ous properties to this substance. We have not had the 
opportunity of testing samples of this class since the 
recognition of the poisonous proteid in those of the 
second class. Four samples of the latter have been tested 
for the poisonous albumin 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.” 
In reference to the poisonous proteids in cheese and other 
articles of food the following interesting questions arise : 
How is the poisoning explained ? Is it not generally sup- 
posed that poisonous proteids are not absorbable from 
mucous membranes? Mitchell and Reichert showed 
that the venom of serpents may be absorbed from mucous 
membranes; especially did they find this to be true of the 
poisonous peptone of the cobra. It may be, however, that 
the bacteria, which are in the cheese and to which the 
formation of the poisonous proteids is due, find their way 62 
BACTERIAL POISONS. 
through the intestinal walls and form their poisonous pro- 
ducts within the spleen and other organs. The fact that 
Novy found the bacteria in the spleen and liver of the 
animals experimented upon confirms this view. 
Poisonous Milk.—In 1885 Vaughan found tyrotoxi- 
con in milk which had stood in a well-stoppered bottle for 
about six months. It was presumed that this milk was, 
when first obtained, normal in composition, but since this 
was not known with certainty, the following experiments 
were made: Several gallon bottles were filled with normal 
milk, tightly closed with glass stoppers, and allowed to 
stand at the ordinary temperature of the room. From time 
to time a bottle was opened and the test for tyrotoxicon was 
made. These tests were followed by negative results until 
about three months after the experiment was begun. Then 
the poison was obtained from one of the bottles. The coagu- 
lated milk was filtered through paper. The filtrate, which 
was colorless and decidedly acid in reaction, was rendered 
feebly alkaline by the addition of potassium hydrate and 
agitated with ether. After separation, the ethereal layer 
was removed with a pipette, passed through a dry filter- 
paper in order to remove a flocculent, white substance which 
floated in it, and then allowed to evaporate spontaneously. 
If necessary, this residue was dissolved in water and again 
extracted with ether. As the ether takes up some water, 
there is usually enough of the latter left after the sponta- 
neous evaporation of the ether to hold the poison in solu- 
tion, and in order to obtain the crystals this aqueous solu- 
tion must be allowed to stand for some hours in vacuo 
over sulphuric acid. 
From one-half gallon of the milk there was obtained 
quite a concentrated aqueous solution of the poison after 
the spontaneous evaporation of the ether. Ten drops of 
this solution placed in the mouth of a small dog, three 
weeks old, caused within a few minutes frothing at the 
mouth, retching, the vomiting of frothy fluid, muscular 
spasms over the abdomen, and after some hours watery 
stools. The next day the dog seemed to have partially 63 
POISONOUS MILK. 
recovered, but was unable to retain any food. This condi- 
tion continuing for two or three days the animal was killed 
with chloroform. No examination of the stomach was 
made. 
In 1886 Newton and Wallace obtained tyrotoxicon 
from milk and studied the conditions under which it forms. 
Their report is of so much value that the greater part of it 
is herewith inserted. 
“ On August 7th twenty-four persons, at one of the hotels 
at Long Branch, were taken ill soon after supper. At 
another hotel, on the same evening, nineteen persons were 
seized with the same form of sickness. From one to four 
hours elapsed between the meal and the first symptoms. 
The symptoms noticed were those of gastro-intestiual irri- 
tation, similar to poisoning by any irritating material— 
that is, nausea, vomiting, cramps, and collapse; a few had 
diarrhoea. Dryness of the throat and burning sensation 
in the oesophagus were prominent symptoms. 
“ While the cause of the sickness was being sought for, 
and one week after the first series of cases, thirty persons 
at another hotel were taken ill with precisely the same 
symptoms as noticed in the first outbreak. 
“ When the news of the outbreak was published one of 
ns immediately set to work, under the authority of the State 
Board of Health, to ascertain the cause of the illness. The 
course of the investigation was about as follows : 
“ The character of the illness indicated, of course, that 
some article of food was the cause, and the first part of our 
task was to single out the one substance that seemed at 
fault. The cooking utensils were also suspected, because 
unclean copper vessels have ofteu caused irritant poisoning. 
Articles of food, such as lobsters, crabs, blue fish, and 
Spanish mackerel, all of which at times, and with some 
persons very susceptible to gastric irritation have produced 
toxic symptoms, were looked for, but it was found that 
none of these had been eaten at the time of the outbreak. 
The cooking vessels were examined, and all were found 
clean and bright, and no evidence of corrosion was pre- 
sented. 64 
BACTERIAL POISONS. 
“Further inquiry revealed the fact that all who had 
been taken ill had used milk in greater or less quantities, 
and that persons who had not partaken of milk escaped 
entirely; corroborative of this, it was ascertained that 
those who had used milk to the exclusion of all other food 
were violently ill. This was prominently noticed in the 
cases of infants fed from the bottle, when nothing but un- 
cooked milk was used. In one case an adult drank about 
a quart of the milk, and was almost immediately seized 
with violent vomiting followed by diarrhoea, and this by 
collapse. Suffice it to say, that we were able to eliminate 
all other articles of food and to decide that the milk was the 
sole cause of the outbreak. 
“ Having been able to determine this, the next step was 
to discover why that article should, in these cases, cause so 
serious a form of sickness. 
“The probable causes which we were to investigate were 
outlined as follows : (1) Some chemical substance, such as 
borax, boric acid, salicylic acid, sodium bicarbonate, sodium 
sulphate, added to preserve the milk or to correct acidity. 
(2) The use of polluted water as an adulterant. (3) Some 
poisonous material accidentally present in the milk. (4) The 
use of milk from diseased cattle. (5) Improper feeding of 
the cattle. (6) The improper care of the milk. (7) The 
development in the milk of some ferment or ptomaine, 
such as tyrotoxicon. 
“At the time of the first outbreak we were unable, un- 
fortunately, to obtain any of the noxious milk, as that un- 
consumed had been destroyed; but at the second outbreak 
a liberal quantity was procured. 
“It was soon ascertained that one dealer had supplied 
all the milk used at the three hotels where the cases of 
sickness had occurred. His name and address having been 
obtained, the next step in the investigation was to inspect 
all the farms, and the cattle thereon, from which the milk 
was taken. We also learned that two deliveries at the 
hotels were made daily, one in the morning and one in the 
evening; that the milk supplied at night was the sole 
cause of the sickness, and that the milk from but oue of POISONOUS MILK. 
65 
the farms was at fault. The cows on this farm were found 
to be in good health, and, besides being at pasture, were 
well fed with bran, middlings, and corn-meal. 
“ So far we had been able to eliminate as causes diseased 
cattle aud improper feeding, and we were then compelled 
to consider the other possible sources of the toxic material. 
“ While the inspection of the farms was being made, the 
analysis of the milk was in progress. The results of this 
showed that no chemical substance had been added to the 
milk, that it was of average composition, that no polluted 
water had been used as a diluent, and that no poisonous 
metals were present. This result left us nothing to con- 
sider but two probable causes : improper care of the milk, 
and the presence of a ferment. 
“As to the former, we soon learned much. The cows 
were milked at the unusual and abnormal hours of mid- 
night and noon, and the noon’s milkiug—that which alone 
was followed by illness—was placed, while hot, in the cans, 
and then, without any attempt at cooling, carted eight miles 
during the warmest part of the day in a very hot month. 
“This practice seemed to us sufficient to make the milk 
unpalatable, if not injurious, for it is well known that when 
fresh milk is closed up in a tight vessel and then deposited 
in a warm place, a very disagreeable odor aud taste are 
developed. Old dairymen speak of the animal heat as an 
entity, the removal of which is necessary in order that the 
milk shall keep well and have a pleasant taste. While we 
do not give this thing a name, we are fully convinced that 
milk should be thoroughly cured by proper chilling and 
aeration before it is transported any distance or sold for 
consumption in towns or cities. 
“ This opinion is based on a study of the methods prev- 
alent among experienced dairymen, who ship large quanti- 
ties of milk to our great cities. The usual practice is to 
allow the milk to stand in open vessels, surrounded by ice 
or cold water, for from eight to twelve hours before trans- 
portation, and when placed on the cars it has a temperature 
of from 50° to 60° F., and is delivered to consumers in a 
perfectly sweet condition. The city of New York receives 66 
BACTERIAL POISONS. 
about 200,000 gallons each day from the surrounding 
country, and much of it brought in by the railroads has 
been on the cars for a time varying from six to twelve 
hours, yet we seldom hear of any of this milk undergoing 
the peculiar form of fermentation set up in the Long 
Branch milk. We may account for this by assuming that 
the proper care of the milk after it was taken from the 
cow, and the low temperature at which it was kept, have 
prevented the formation of any ferment; this opinion 
seems to be endorsed by all dairymen and managers of 
large creameries with whom we have consulted. They all 
agree in stating that milk maintained at a low temperature 
can be kept sweet and in good condition for many days. 
“ We have dwelt on this branch of our topic somewhat 
extensively, because we are fully persuaded that the im- 
proper care of the milk had much to do with the illness it 
produced. 
“ The results of our inquiry having revealed so much, 
we next attempted to isolate some substance from the 
poisonous milk, in order that the proof might be more 
evident. A quantity of the milk that had caused sickness 
in the second outbreak was allowed to coagulate, was then 
thrown on a coarse filter, and the filtrate collected. This 
latter was highly acid, and was made slightly alkaline by 
the addition of potassium hydrate. This alkaline filtrate 
was now agitated with an equal volume of pure, dry ether, 
and allowed to stand for several hours, when the ethereal 
layer was drawn off by means of a pipette. Fresh ether 
was added to the residuum, then agitated, and, when sepa- 
rated, was drawn off and added to the first ethereal 
extract. This was now allowed to evaporate spontane- 
ously, and the residue, which seemed to contain a small 
amount of fat, was treated with distilled water and filtered, 
the filtrate treated with ether, the ethereal solution drawn 
off and allowed to evaporate, when we obtained a mass of 
needle-shaped crystals. This crystalline substance gave a 
blue color with potassium ferricyanide and ferric chloride, 
and reduced iodic acid. The crystals, when placed on the 
tongue, gave a burning sensation. A portion of the crys- POISONOUS MILK. 
67 
tals was mixed with milk and fed to a cat, when, in the 
course of half an hour, the animal was seized with retching 
and vomiting, and was soon in a condition of collapse, from 
which it recovered in a few hours. 
“We are justified in assuming, after weighing well all 
the facts ascertained in the investigation, that the sickness 
at Long Branch was caused by poisonous milk, and that 
the toxic material was tyrotoxicon. 
“ The production of this substance was no doubt due to 
the improper management of the milk—that is, too long a 
time was allowed to elapse between the milking and the 
cooling of the milk, the latter not being attended to until 
the milk was delivered to the hotel; whereas, if the milk 
had been cooled immediately after it was drawn from the 
cows, fermentation would not have ensued, and the result- 
ing material, tyrotoxicon, would not have been produced.” 
In the same year, Schearer found the same poison in 
the milk used by, and the vomited matter of, persons made 
sick at a hotel at Corning, Iowa. 
In 1887, Firtii, an English army surgeon stationed in 
India, reported an outbreak of milk poisoning among the 
soldiers of his garrison. From the milk he separated, by 
Vaughan’s method, tyrotoxicon. He also obtained tyro- 
toxicon from milk which had been kept for some months 
in stoppered bottles, as had been previously done by 
Vaughan. (See page 62.) 
In 1887, Mesic and Vaughan observed four cases of 
milk poisoning, three of which terminated fatally, and 
Novy and Vaughan obtained tyrotoxicon from the milk, 
and from the contents of the intestine in one of the fatal 
cases. Vaughan reports these cases as follows : 
“ September 23, 1887, I was visited by Dr. A. G. Mesic, 
of Milan, Michigan, who informed me that he had four 
members of a family under his charge, all of whom were 
seriously ill with peculiar symptoms which he believed to 
be caused by tyrotoxicon. Since Dr. Mesic has written 
out for me the history of these cases, I will insert his report 
in full, as follows : 
“ ‘ Saturday, September 17, while passing the residence 68 
BACTERIAL POISONS. 
of S. II. Evans, a respectable farmer, I was called in to see 
him. I found him—a man of about fifty years, spare 
and muscular—vomiting severely, with flushed face, but 
with a temperature of 96° F. There was marked throb- 
bing of the abdominal aorta; the tongue had a white, 
heavy coating, and the breathing was very labored. I set 
to work with the ordinary remedies to allay the vomiting, 
which had already continued for some hours. The vomited 
matters were colored with bile. Pupils were dilated, and 
a rash resembling that of scarlatina, but coarser, covered 
the chest, forearms, and legs below the knees, while the 
abdomen and thighs remained unaffected. As the bowels 
had not been moved since the beginning of the attack, I 
administered a purgative dose of calomel with a little podo- 
phyllin and rhubarb. On Sunday a small stool resulted. 
During that day and night, and the following day, the 
retching and vomiting continued. Small doses of carbolic 
acid seemed to give the most relief. After the movement 
of the bowels the symptoms were somewhat more prom- 
ising ; but a heavy and unfavorable stupor was observable 
and persistent. 
“ ‘ On Sunday the coating of the tongue remained very 
thick, and had changed to a dark brown color. At first I 
thought that his symptoms indicated a depressed condition, 
which I had known in one instance to precede typhoid 
fever. However, after a few days, I concluded that I 
must look for the cause of the condition among the poi- 
sons ; but I could think of no one poison which would 
be likely to produce all the symptoms observed. During 
Monday, Tuesday, and Wednesday, there was but little 
change, and the treatment was continued. 
“ ‘ On Thursday morning I found the son Arthur, a lad 
of eighteen years, strong and vigorous, suffering with the 
same symptoms, only in a more violent form. After 
supper on Wednesday evening he was taken with nausea 
aud vomiting. He had no rash, but the symptoms were 
otherwise identical with those of the father, except in being 
more severe. I gave a cathartic, which acted only slightly. 
“ ‘ Aft my evening visit I found Mrs, Evans, a lady of POISONOUS MILK. 
69 
about forty-five, previously in good health, with the same 
symptoms. In this case the stupor was more marked from 
the first. I was unable at any time to obtain any cathartic 
action in this case. Copious enemata of warm water were 
used, but succeeded only in washing some hardened lumps 
from the rectum. By this time I had concluded that the 
poison was most likely tyrotoxicon. 
“‘On Friday morning the only remaining member of 
the family at home, Miss Alma, sixteen years of age, was 
affected in the same way as the others. On that day I 
went to Ann Arbor, and gave a history of the cases so far 
to Dr. Vaughan, who, from the symptoms, thought that 
my diagnosis was most probably correct, and he advised 
with me as to treatment, which I carried out. I gave two 
grains of sodium salicylate every four hours, and used small 
doses of the tonics and stimulants, quinine, mix vomica, 
digitalis, whiskey, and the aromatic spirits of ammonia. 
On Saturday the symptoms in all remained unimproved, 
and in the mother and son the stupor and labored breath- 
ing grew more marked. 
“ ‘ On Sunday, I again went to Ann Arbor, and brought 
Dr. Vaughan with me to see the patients. The tempera- 
ture of the mother on Sunday was as low as 94° F., and 
that of the son 95° F. Dr. Vaughan agreed with me as to 
diagnosis and treatment. Sunday evening the patients 
were all removed to the house of a neighbor, about forty 
rods distant (the reasons for this will be given later). Dr. 
Vaughan and I both expressed the fear that the mother, 
and possibly the son, would not live through the night. 
Both of these rapidly grew worse, and the son died at 7.45 
A.M. and the mother at 4 P.M., Monday. 
“ 1 During Monday the daughter rapidly grew worse, and 
at the time of her mother’s death could not be aroused, and 
practically she remained unconscious from that time on. 
The father was very weak, but retained his consciousness 
all the time. Convulsive movements of the limbs had 
been noticed in the son, but not in the mother. These 
now became more marked in the daughter, who remained 70 
BACTERIAL POISONS. 
in the heavy stupor, with labored breathing, until 5 p.m. 
Thursday, when she died. 
“‘ Mr. Evans has slowly improved, and now, October 
18th, is able to walk about the room. The sodium sali- 
cylate, even in the small doses used, seemed to cause severe 
headache; so apparent was this that the drug was discon- 
tinued, and drop doses of amyl nitrite, given every hour, 
seemed to relieve the pain in the head. His temperature 
remained below the normal until Thursday, October 14th, 
when it reached the normal. After this it was found once 
as high as 99.5° F., then 99° F., then again normal, where 
it remains. 
“ ‘All complained of a burning constriction in the throat, 
and difficulty in swallowing, and all, as long as they were 
conscious, frequently called for ice. In all the pulse was 
rapid and feeble, and death -seemed to result from failure 
of the heart. Those who died voided urine involuntarily, 
while Mr. Evans passed small quantities frequently, and 
for this buchu and uva ursa were given. During his con- 
valescence small doses of morphine were given, as he was 
unable to sleep, and became very restless. He is now 
taking teaspoonful doses of the elixir of calisaya and iron 
every four hours/ 
“ As stated above by Dr. Mesic, I first saw these patients 
Sunday, September 25th. On a sofa iit the room we found 
the daughter, Alma. She had been vomiting during the 
day, and seemed much exhausted. She was not inclined to 
talk, and seemed to be in a stupor, though when spoken to 
she responded rationally. Her pupils were slightly dilated, 
her tongue coated, her pulse 120 and weak, her face flushed, 
and a violent throbbing could be felt over the abdomen, 
which was retracted. Her temperature was 96° F. 
“ In another room were the father, mother, and son, two 
of them dying. The father was rational, and talked with 
some freedom when I asked as to the kind of food they 
had been eating, etc. His pupils were normal. Ilis face 
could not be said to present any peculiar feature. His 
pulse was rapid, breathing somewhat labored, and the 
throbbing of the abdominal aorta was plainly felt. The 71 
POISONOUS MILK. 
abdomen was retracted, and there was no pain on pressure. 
He complained of a burning constriction of the throat, 
swallowed with difficulty, and said that his throat and 
stomach felt as though they were on fire. 
“The mother lay perfectly still with eyelids closed, as if 
in a deep sleep. Her pulse was rapid, her face had a livid 
flush, her breathing was about 35 per minute, and labored. 
The skin was cool, but neither abnormally moist nor 
specially dry and harsh. She could not be aroused. In 
fact, she was comatose. 
“ The son rolled uneasily from one side of the bed to the 
other. His breathing, also, was very labored. His eyelids 
were closed, and the pupils were markedly dilated—did not 
respond to light. He could not be aroused. In mother 
and son, as well as in father and daughter, the abdomen 
was retracted, and the throbbing of the abdominal aorta 
Avas easily felt. 
“Noav, to what were these symptoms due? They were 
certainly those of some poison. Dr. Mesic had brought 
me some of the vomited matter, which I tested thoroughly 
for mineral poisons, with negative results. The symptoms 
certainly Avere not those of morphine, strychnine, digitalis, 
or aconite. They did have some resemblance to those of 
belladonna, but yet they were not the symptoms of bella- 
donna. The pupils Avere not as Avidely dilated as they 
would be in belladonna poisoning. There Avas in none of 
these persons the active delirium of belladonna poisoning. 
There was no picking at the clothing, no grasping of imag- 
inary objects in the air, no hallucinations of vision. Surely 
it could not be any vegetable alkaloid Avith which I Avas 
familiar. 
“ On the other hand, we know that nausea, vomiting, 
headache, dilatation of the pupil, rapid pulse, heavy breath- 
ing, constipation, and great prostration, with stupor, do 
occur in cases of poisoning with certain ptomaines. There- 
fore Ave began to look for conditions Avhich Avould be favor- 
able for the production of putrefactive alkaloids. These 
conditions Ave Avere not long in finding. 
“ The family, Avhich consisted of the four persons sick, and 72 
BACTERIAL POISONS. 
of a daughter about twenty years of age, who was away 
from home at the time when the others were taken ill, 
and for some months before that time, was evidently a 
tidy one. This was shown by their personal appear- 
ance, and by the clothing and bedding. But the house 
in which they lived was very old, and very much 
decayed. Mr. Evans had purchased the farm six years 
ago; and for some three years past, at least, they had been 
troubled every now and then, one or more of the family, 
with nausea and vomiting, followed by more or less prostra- 
tion. But in no instance, up to the present illness, had the 
symptoms been sufficient to cause them to summon a physi- 
cian. The family had worked hard in order to pay for the 
farm, and had determined to make the old house do until 
they were out of debt. Even before this family had moved 
to the farm, the house had tjeen known among the neigh- 
bors as an unhealthy one, and there had been much sick- 
ness and a number of deaths among its former tenants. 
“ The house is a frame one, and one of the neighbors said 
to me that it was an old house when he came to the neigh- 
borhood thirty-seven years ago. It consists of two rooms 
on the ground-floor, with attic rooms above. The frame 
rests upon four large logs or sills, which lie directly upon 
the ground, and are thoroughly rotten. There is no cellar 
under any part of the house. From the front, at least, the 
surface slopes toward the house, and the rain-water runs 
under it. In the floor of one room a trap-door had been 
placed, and directly under this a small excavation had been 
made for the purpose of collecting the rain-water when it 
accumulated under the house. Although this pit was dry 
at the time of our examination, its sides and bottom were 
marked with cray-fish holes, showing that water had stood 
in it. The floor was laid of unjointed boards, and every 
time that it was swept much of the filth fell through the 
cracks, and every time that the tidy housewife scoured and 
mopped the floor, the water, carrying with it the filth, ran 
through the crevices, and thus the conditions most favorable 
for putrefactive changes were brought into existence and 
maintained. POISONOUS MILK. 
73 
“ One corner of one of the rooms had been transformed 
into a small room, or buttery, as it was called, and in this, 
on shelves, the food was kept. On account of the more 
frequent scouring demanded by that part of the floor 
enclosed in this buttery, the boards had rotted away, and a 
second layer of boards had been placed over the original 
floor. Between these two floors we found a great mass of 
moist, decomposing matter, the accumulations of years, 
which the broom could not reach. When this floor was 
taken up, a peculiar, nauseating odor was observable, and 
was sufficient to produce nausea and vomiting in one of the 
persons engaged in the examination. Some of the dirt 
from beneath the floor, and some of that which had accumu- 
lated beneath the boards in the buttery, were taken for 
further study. 
“ The condition of the house was supposed to be unfavor- 
able to the patients, and for this reason they were moved, 
as Dr. Mesic has stated, to the house of a neighbor. Of 
course, thorough examination of the house was not made 
until the patients had been removed. 
“ Special inquiry was now made concerning the food used 
by this family. They had been living very simply. They 
lived upon bread, butter, milk, and potatoes, with coffee 
and ripe fruit. They had eaten no canned foods for months. 
They ate but little meat. Occasionally a chicken was killed 
and served, and rarely, some fresh meat was obtained from 
the village. During the week in which they were taken ill, 
all the meat used consisted of slices from a piece of bacon, 
the only meat which was kept in the house, and a chicken. 
None of the latter remained, but the bacon was examined. 
It seemed in perfect condition, and contained no trichina;. 
Moreover, as has been seen from the history of the cases, 
all the members of the family were not made sick by any 
one meal, but the opportunity of obtaining the poison must 
have been present for some time. Moreover, the fact that 
previous similar, but less severe, attacks had occurred at 
intervals for the past three years, convinced us that the 
poison must owe its origin to some long-existing condition. 
“ The drinking-water supply was also investigated. The 74 
BACTERIAL POISONS. 
water was obtained from a shallow well, and some of it was 
taken for analysis. But several families had for years used 
water from this well, and had remained healthy. 
“ The milk used by the family was studied. Of course, 
we could get none of that which had been used before the 
members of the family were stricken down. As soon as he 
made the diagnosis of tyrotoxicon poisoning, Dr. Mesic 
ordered the discontinuance of the use of milk, not only with 
the sick, but he forbade the daughter, who had returned, 
and any of the visitors using it. Mr. Evans owned four 
milch cows, and they were supplied with fair pasturage and 
abundant water. The greater part of the milk was placed 
in tin cans which were set in a wooden trough in the yard, 
and surrounded by cold water. The covers to the cans 
were arranged so that the air could have free access to the 
milk, and were left in this position until the milk was 
thoroughly cooled. Indeed, the cans were furnished by a 
creamery company, which followed the directions which I 
have previously given for the care of milk. On his first 
visit to me, Dr. Mesic brought some of the milk from one 
of these cans. This I examined, but failed to find tyro- 
toxicon in it. 
“ However, the family did not drink any of the milk from 
the cans. That which they did use was kept in the buttery 
which I have described. Here it stood upon a shelf, and 
some members of the family, at least, were in the habit of 
drinking from it between meals. This was especially true, 
it is said, of the son. He would frequently come from his 
work in the fields, go into the buttery and drink a glass or 
more of the milk. Mr. Evans states that he frequently 
observed that the taste of the milk was not pleasant. On 
my first visit to the premises I advised that some of the 
milk should be taken from the cans, allowed to stand in the 
buttery over night, and be sent to me the next day. This 
was done, and in this milk we found tyrotoxicon, not only 
by the employment of chemical tests, but by poisoning a 
kitten with it. 
“On the death of the mother and son, Dr. Mesic asked for 
a post-mortem, but the friends objected, and the undertaker POISONOUS MILK. 
75 
used an arsenical embalming fluid, so that, although consent 
was subsequently obtained, it was decided that the exami- 
nation would be so vitiated as to be worthless. On the 
death of the daughter, the coroner summoned a jury and 
held an inquest. The post-mortem was conducted by I)r. 
George A. Hendricks, in the presence of the jury and 
several physicians who had been invited. Dr. Hendricks 
has kindly furnished me with his report, which I present 
here in full: 
“ The autopsy was held fifteen hours after death. The 
abdominal viscera were first examined. The great omen- 
tum was small, in normal position, covering the small 
intestine. The small intestine was moderately distended 
with flatus. The jejunum was ashy-green in color; the 
ileum purplish-green. About eighteen inches from the ter- 
mination of the ileum was found a diverticulum two inches 
in length. The small intestine contained very little ali- 
mentary matter. The vermiform appendix was free, con- 
tained some small fecal lumps, and showed no evidence of 
inflammation. The caecum, ascending, transverse, and 
descending colon were empty and their circular fibres were 
tightly constricted, except at intervals where the intestine 
was distended with gas. The sigmoid flexure was moder- 
ately distended with gas, and the rectum contained small 
bits of fecal matter. The stomach was somewhat contracted 
and lay wholly upon the left side of the median line. It 
contained a few ounces of fluid. Its extremities were ligated 
and the organ removed. The mucous membrane of the 
stomach and intestine were not examined until they reached 
the chemist. The duodenum was distended with flatus. 
The liver was normal in size and appearance. The gall- 
bladder contained about one ounce of bile. The spleen was 
normal. One-half ounce of fluid deeply stained with blood 
was found in Douglas’s cul-de-sac. The uterus, Fallopian 
tubes, and ovaries were deeply congested. The left ovary 
was enlarged and presented on its posterior surface a hemor- 
rhagic spot, oval, about one-half line in length, and several 
other less distinct ones. The right ovary was normal in 
size and showed numerous Graafian scars. The ureters 76 
BACTERIAL POISONS. 
and bladder were normal; the latter contained a small 
amount of urine. The peritoneum, pancreas, and kidneys 
were perfectly normal. 
“The thoracic cavity was next opened. The lungs were 
normal; there was about one-half ounce of free serum in 
the left pleural cavity; none in the right. Pericardium 
normal; right auricle in diastole; left auricle and both 
ventricles in systole. 
“ The dura mater showed venous congestion ; the arach- 
noid, normal ; the pia mater, congested. On the surface 
of the centrum ovale, small drops of blood oozed from the 
divided vessels. The large veins of the velum interposi- 
tion were distended. Third and fourth ventricles were 
slightly distended with serous fluid, but the walls were 
normal. There seemed to be slight softening of the optic 
thalami. The sub-arachnoid fluid was about twice the nor- 
mal quantity. 
“ On examination of the mucous membrane of the 
stomach and intestine in the presence of the chemist, Prof. 
A. B. Prescott,’nothing abnormal could be found. The 
membrane was stained with bile, but there was not the 
slightest redness. The solitary glands were distinct, but 
not at all inflamed. Peyer’s patches were normal. 
“It will be seen that there existed no lesion which would 
account for the death. The venous congestion observed in 
the brain would follow from failure of the heart. 
“ Some of the post-mortem appearances bore a striking 
resemblance to those which I had observed in cats poi- 
soned with tyrotoxicon. This was especially noticeable in 
the condition of the mucous membrane of the stomach and 
intestine. Tyrotoxicon produces the symptoms of a gas- 
trointestinal irritant, but not the lesions. The contraction 
of the circular fibres of the intestine, which undoubtedly 
caused the constipation, I had also observed in cats that 
died from tyrotoxicon poisoning without either vomiting 
or stool.1 The action of this poison upon the stomach and 
1 Marsh reports a case in which the symptoms resembled very closely 
those of rapidly perforating typhlitis, but the post-mortem examination 
showed absolutely no evidence of this disease or of peritonitis. In fact the POISONOUS MILK. 
77 
intestine must be through the nervous system. Small 
doses cause both vomiting and purging, while after large 
doses vomiting may be impossible, and obstinate constipa- 
tion may exist. Both the vomiting and purging after 
small doses are undoubtedly due in part to increased 
activity of the circular fibres of the muscular coats, induced 
through the nerves; and the inability to vomit, and the 
constipation, one or both of which may be observed after 
large doses of the poison, are due to spasm of the same 
muscles, induced in the same manner. 
“ Prof. A. B. Prescott was requested by the coroner to 
analyze the material for mineral and vegetable poisons. 
He made analyses of the stomach and part of its contents, 
and a portion of the liver. His results were wholly nega- 
tive. 
“Novy tested a cold-water extract of the finely divided 
intestine for ptomaines. The fluid, which was acid in 
reaction, was filtered, then neutralized with sodium bi- 
carbonate, and shaken with ether. The ether, after 
separation, was removed, and allowed to evaporate spon- 
taneously. The residue was dissolved in water, and 
extracted again with ether. This ether residue gave the 
chemical reactions for tyrotoxicon, aud a portion of it was 
administered to a kitten about two months old. Within 
half an hour after the administration the kitten began to 
retch, and soon it vomited. Within the next three hours 
it was noticed to vomit as many as five times. The breath- 
ing became rapid and labored. The animal sat with its 
head down, aud seemed greatly prostrated. The pupils 
were examined, but could not be said to be dilated. There 
was no purging. The retching and heavy breathing, with 
evidences of prostration, continued more or less marked 
for two days, after which the animal slowly improved. 
“A quantity of fresh milk was divided into five por- 
tions of one quart each, placed in quart bottles which had 
only abnormality found in the intestines consisted of the contraction ot the 
circular fibres of the transverse and descending colon. Marsh believes 
that this was a case of ptomaine poisoning. 78 
BACTERIAL POISONS. 
been thoroughly cleansed, and treated in the following 
manner: 
“No. 1 consisted of the milk only, and was employed* 
as a control test. 
. “ No. 2 was mixed with a drachm of vomited matter. 
“No. 3 was treated with a portion of the contents of 
the stomach. 
“ No. 4 was treated with an aqueous extract of the in- 
testine. 
“ No. 5 was treated with a small portion of the soil 
which had been taken from the floor of the buttery, stirred 
up with water. 
“ These bottles were placed in an air-bath, and kept at a 
temperature of from 25° to 30° C. for twenty-four hours. 
Then each was tested for ptomaines. No. 1 yielded no 
tyrotoxicon, while all of the others contained this poison. 
The tests were both chemical and physiological. All of 
the samples yielded a non-poisonous base when treated 
according to Brieger’s method, and the same substance was 
obtained from perfectly fresh milk. It is most probably 
formed by the action of the heat and reagents employed in 
this method. This base was obtained in crystalline form, 
and several portions of it were administered to kittens 
without any effect. The further study of this body will 
be of interest to toxicologists, because it gives many of the 
general alkaloidal reactions. At first we supposed it to be 
Brieger’s neuridine, and this supposition may still be cor- 
rect, but, as we obtained it, it gave some reactions which 
are not given by neuridine. Further investigations will 
be made on this point. 
“ Tyrotoxicon was obtained from the filtered milk by 
two methods: (1) The one which we have previously used, 
and which consists in neutralizing the filtered milk with 
sodium bicarbonate, and extracting with ether. That por- 
tion of the poison employed in the physiological tests was 
obtained in this way, and in order to be sure that no poison 
came from the ether, the extract from the milk to which 
nothing had been added was given to a kitten, and was 
found to produce no effect, (2) The filtrate from the milk POISONOUS ICE-CREAM. 
79 
was heated to 70° C. (158° F.) (tyrotoxicou decomposes at 
91° C. (195.8° F.)) for some minutes, and filtered. This 
filtrate, which was perfectly clear, was treated with a small 
quantity of nitric acid in order to convert the tyrotoxicou 
into a nitrate, then pure potassium hydrate in the solid 
form was added until the solution was strongly alkaline. 
This solution was concentrated so far as it could be on the 
water-bath. (The potassium compound of tyrotoxicon is 
not decomposed below 130° C. (234° F.).) The dark 
brown residue, after cooling, was examined with the micro- 
scope and found to contain the crystalline plates of tyro- 
toxicon-potassium hydrate, along with the prisms of potas- 
sium nitrate. The former was separated from the latter 
by extraction with absolute alcohol and filtration. The 
alcohol was evaporated to dryness on the water-bath, and 
the residue again extracted with absolute alcohol. From 
this alcoholic solution tyrotoxicon was precipitated with 
ether. The precipitate was decomposed by adding acetic 
acid and heating, the tyrotoxicon being broken up into 
nitrogen and phenol. The phenol was recognized by pre- 
cipitation with bromine water, and by other well-known 
tests. 
“ On October 8th, the coroner’s inquest, which had been 
adjourned after the post-mortem in order to await the re- 
sults of the analysis, was resumed, and after hearing the 
testimony in accordance with the above stated facts, the 
jury returned a verdict of death from poisoning with tyro- 
toxicon.” 
Camman reports twenty-three cases of milk poisoning 
which he attributes to tyrotoxicon, although this poison 
could not be found in the milk. It may be that the active 
agent present belongs to the bacterial proteids. 
Kinnicutt has isolated tyrotoxicon from milk which 
had been kept for some hours in an unclean vessel. 
Poisonous Ice-cream.—In 1886, Vaughan and 
Novy obtained tyrotoxicon from a cream which had 
seriously affected many person at Lawton, Michigan. 
Vanilla had been used for flavoring, and it was supposed 80 
BACTERIAL POISONS. 
that the ill-effects were due to the flavoring. This belief 
was strengthened by the fact that a portion of the custard 
was flavored with lemon, and the lemon cream did not 
affect any one unpleasantly. Fortunately some of the 
vanilla extract remained in the bottle from which the fla- 
voring for the ice-cream had been taken, and this was for- 
warded to the chemists. Each of the experimenters took 
at first thirty drops of the vanilla extract, and no ill-effects 
following this, one of them took two teaspoonfuls more, 
with no results. This proved the uon-poisonous nature of 
the vanilla more satisfactorily than could have been done 
by a chemical analysis. 
Later, it was found that that portion of the custard 
which had been flavored with lemon was frozen immedi- 
ately ; while that portion which was flavored with vanilla 
and which proved to be poisonous, was allowed to stand 
for some hours in a building, which is described as follows 
by a resident of the village: 
“ The cream was frozen in the back end of an old 
wooden building on Main Street. It is surrounded by 
shade, has no underpinning, and the sills have settled into 
the ground. There are no eve-troughs, and all the water 
falling from the roof runs under the building, the streets 
on two sides having been raised since the construction of 
the house. The building had been unoccupied for a num- 
ber of months, consequently had had no ventilation, and 
what is worse, the back end (where the cream was frozen) 
was last used as a meat market. The cream which was 
affected was that portion last frozen ; consequently it stood 
in an atmosphere like that of a privy vault for upward of 
an hour and a half or two hours before being frozen.” 
The symptoms observed in these cases are given by Dr. 
Mopitt as follows: 
“About two hours after eating the cream every one was 
taken with severe vomiting, and after from one to six 
hours later with purging. The vomit was of a soapy char- 
acter, and the stools watery and frothy. There was some 
griping of the stomach and abdomen, with severe occipital 
headache, excruciating backache, and bone pains all over, POISONOUS ICE-CREAM. 
81 
especially marked in the extremities. The vomiting lasted 
from two to three hours, then gradually subsided, and 
everybody felt stretchy, and yawned in spite of all resist- 
ance. The throats of all were cedematous. One or two 
were stupefied; others were cold and experienced some 
muscular spasms. A numb feeling, with dizziness and 
momentary loss of consciousness, was complained of by 
some. Temperature was normal, and pulse from 90 to 
120. Tongue dry and chapped. All were thirsty after 
the vomiting subsided, and called for cold water, which 
was allowed in small quantities, with no bad results. 
After getting out no one of the victims was able to be in 
the hot sun for several days, and even yet (about ten days 
after the poisoning) the heat affects myself. I attended 
twelve persons, besides being sick myself, and all were 
affected in pretty much the same way. Several complain 
yet of inability to retain food on the stomach without dis- 
tressing them. The man who made the cream took a tea- 
spoouful of it, and he vomited the same as those who took 
a whole dish, but not so often or for so long a time. All 
are affected with an irresistible desire to sleep, which can 
scarcely be overcome. Even yet, some of us feel that 
drowsy condition, with occasional occipital headache.” 
The tyrotoxicon obtained from this cream was adminis- 
tered to a kitten about two months old. Within ten 
minutes the cat began to retch and soon it vomited. This 
retching and vomiting continued for two hours, during 
which the animal was under observation, and the next 
morning it was observed that the animal had passed several 
watery stools. After this, although the kitten could walk 
about the room, it was unable to retain any food. Several 
times it was observed to lap a little milk, but on doing so 
it would immediately begin to retch and vomit. Even cold 
water produced this effect. This condition continuing, 
after three days the animal was placed under ether and its 
abdominal organs examined. Marked inflammation of the 
stomach was supposed to be indicated by the symptoms, 
but the examination revealed the stomach and small intes- 
tine filled with a frothy, serous fluid, such as had formed 82 
BACTERIAL POISONS. 
a portion of the vomited matter, and the mucous membrane 
very white and soft. There was not the slightest redness 
anywhere. The liver and other abdominal organs seemed 
normal. 
A bit of the solid portion of this cream was added to 
some normal milk, which, by the addition of eggs and 
sugar, was made into a custard. The custard was allowed 
to stand for three hours in a warm room, after which it 
was kept in an ice-box until submitted to chemical analysis. 
In this tyrotoxicon was also found. 
Tyrotoxicon has since been found in some chocolate 
cream which poisoned persons at Geneva, N. Y., and in 
lemon cream from Amboy, Ohio. 
Sciiearer reports the finding of tyrotoxicon in both 
vanilla and lemon ice-cream which made many sick at 
Nugent, Iowa. 
Allaben reports poisoning with lemon cream, and 
makes the following interesting statements concerning it: 
“ I would first say July 4, 5, and 6 were very warm. 
Monday evening, July 5, the custards were cooked, made 
from Monday morning’s cream and Monday night’s milk, 
boiled in a tin pan that had the bright tin worn off. It 
was noticed that one pan of cream was not sweet, but 
thinking it would make no difference, it was used; the 
freezers were thoroughly cleaned and scalded, and the 
custards put in the same evening while hot; the cream was 
frozen Tuesday afternoon, having stood in the freezers 
since the night before, when the weather was very warm.” 
No analysis of this cream was made, but the symptoms 
agree with those of tyrotoxicon poisoning. 
Welford observed several cases of poisoning from 
custard flavored with lemon. These custards were tested 
for mineral poisons, with negative results. 
Morrow has put forth the claim that ice-cream poison- 
ing is solely due to artificially prepared vanillin, which is, 
according to his statement, used instead of vanilla extract, 
but the facts stated above concerning poisoning with creams 
in which other flavors had been used contradict this claim. 
Moreover, Gibson has shown the utter absurdity of the POISONOUS MEAL AND BREAD. 
83 
claim, inasmuch as he calculates from the amount of flavor- 
ing ordinarily used in ice-cream, that in order to produce 
the toxic symptoms observed, the flavoring must be ten 
times as poisonous as pure strychnine. 
Bartley suggests that poisonous cream sometimes 
results from the use in its manufacture of poor or putrid 
gelatin. This is highly probable, and with the gelatin 
the germs of putrefaction may be added to the milk. 
Poisonous Meal and Bread.—Reference has already 
been made to the fact that the peasants in certain parts of’ 
Italy are frequently poisoned by eating mouldy corn-meal. 
As has also been stated, Lombroso and others have ob- 
tained from this meal ptomaines, some of which give the 
same color reaction as strychnine. In 1886, Ladd suc- 
ceeded in isolating from “ heated ” corn-meal a ptomaine 
which forms in urea-like crystals. The quantity was not 
sufficient for an ultimate analysis, and the physiological 
action has not been studied. Poisoning from decomposed 
and mouldy bread is not unknown. CHAPTER IV. 
GENERAL CONSIDERATIONS OF THE RELATION OF 
BACTERIAL POISONS TO INFECTIOUS DISEASES. 
The majority of diseases may be grouped from an etio- 
logical standpoint into the following classes: (1) Trau- 
matic ; (2) infectious; (3) autogenous ; and (4) neurotic. 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 en- 
trance 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 indi- 
vidual 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 influences that it dies without producing any appre- 
ciable effects; or all may be infected, but with difference in 
degree, as is evidenced by variation in symptoms, in the 
length of time that tin's 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. RELATION TO INFECTIOUS DISEASES. 
85 
Charrin and Roger have shown that white rats, 
which are naturally immune to anthrax, become susceptible 
when fatigued by being kept ou a small tread-mill. Eleven 
rats were inoculated with an anthrax culture; five of these 
which were allowed to rest in the cage manifested no symp- 
toms of the disease, while six which were placed on the 
tread-mill developed the disease and died within from 
twenty-four to thirty hours. The bacilli were found in 
the liver and spleen of those which died ; and guinea-pigs 
inoculated with these germs died. The influence of the 
condition of health on susceptibility to the infectious dis- 
eases has also been shown by Leo, who found that mice 
which are naturally insusceptible to glanders, become highly 
susceptible when they are rendered diabetic by the adminis- 
tration of phloridzin. 
That some neurotic affections originate from traumatism 
we know. That others of this class are largely due to mal- 
nutrition accompanied by improper metabolism or insuffi- 
cient elimination, or, in other words, are to some extent 
autogenous, all believe. Understanding, then, that the 
above classification does not attempt a sharp and marked 
differentiation of the causes of disease, we will now give 
our attention to a consideration of the chemical factors in 
the causation of the infectious diseases, and of the trau- 
matic, autogenous and neurotic, in so far as these are influ- 
enced by infection. 
Recognizing the fact that germs do bear a causal relation 
to some diseases, the question arises, How do these organ- 
isms produce disease? In what way does the bacillus 
anthracis, for instance, induce the symptoms of the disease 
and death? Many answers to this question have been 
offered. Some of the most important of these are as fol- 
lows : 
1. It was first suggested by Bollinger that apoplecti- 
form anthrax is due to deoxidation of the blood by the 
bacilli. These germs are aerobic, and were supposed to 
deprive the red blood-corpuscles of their oxygen. This 
theory was suggested most probably by the resemblance of 
the symptoms to those of carbonic acid poisoning. The 86 
BACTERIAL POISONS. 
most prominent of these symptoms are dyspnoea, cyanosis, 
convulsions, dilated pupils, subnormal temperature, and, 
in general, the phenomena of asphyxia. Moreover, post- 
mortem examination reveals conditions similar to those 
observed after death by deprivation of oxygen. The veins 
are distended, the blood is dark and thick, the parenchy- 
matous organs are cyanotic, and the lungs hyperaemic. 
Bollinger compared this form of anthrax to poisoning 
with hydrocyanic acid, which was then believed to produce 
fatal results by robbing the blood of its oxygen. 
This theory was supported by the observations of Szpil- 
mann, who found that while the putrefactive bacteria are 
destroyed by ozone, the bacillus anthracis thrives and mul- 
tiplies in this gas. 
This theory pre-supposed a large number of bacilli in the 
blood, and this accorded with the estimate of Davaine, 
which placed the number at from eight to ten million in a 
single drop. But more extended and careful observation 
showed that the blood of animals dead from anthrax is 
often very poor in bacilli. Virchow reported cases of 
this kind. Bollinger himself found the bacilli often 
confined to certain organs and not abundant in the blood. 
Then Siedamgrotzky counted the organisms in the blood 
in various cases and found not only that the estimate made 
by Davaine is too large, but that in many instances the 
number present in the blood is small. Joffroy found in 
some of his inoculation experiments that the animals died 
before any bacilli appeared in the blood. These and other 
investigations of similar character began to cause workers 
in this field of research to doubt the truth of the theory of 
Bollinger, and these doubts were soon converted into 
positive evidence against it. Pasteur, in support of the 
theory, reported that birds were not susceptible to anthrax, 
and lie accounted for this by supposing that the blood 
corpuscles in birds do not part with their oxygen readily. 
However, it was shown by Oemler and Feser that the 
learned Frenchman had generalized from limited data, and 
that many birds are especially susceptible to the disease. 
Oemler found that the blood even when rich in bacilli relation to infectious diseases. 
87 
still possesses the bright-red color of oxy-haemoglobin. 
Toepper and Roloff reported cases of apoplectiform 
anthrax in which there was no difficulty in respiration. 
Toussaint caused animals which had been inoculated with 
the anthrax bacillus to breathe air containing a large 
volume of oxygen, and found that this did not modify the 
symptoms or retard death. Finally, Nencki determined 
the amount of physiological oxidation going on in the bodies 
of animals sick with anthrax by estimating the amount of 
phenol excreted after the administration of one gramme of 
benzol, and found that the oxidation of the benzol was not 
diminished by the disease. Thus, the theory that germs 
destroy life by depriving the blood of its oxygen has been 
found not to be true for anthrax, and it' not true for 
anthrax, certainly it cannot be for any other known disease. 
The bacillus anthracis is, as has been stated, aerobic, while 
most of the pathogenic bacteria arc anaerobic—that is, they 
live in the absence of oxygen. This element is not neces- 
sary to their existence, and, indeed, when present in large 
amount, it is fatal to them. Moreover, in many diseases, 
the bacteria are not found in the blood at all. Lastly, the 
symptoms of these diseases are not those of asphyxia. These 
facts have caused all bacteriologists to acknowlege that this 
theory is not the right one. 
2. If a properly stained section of a kidney taken from 
a guinea-pig, which has been inoculated with the bacillus 
anthracis, be examined under a microscope, the bacilli will 
be found to be present in such large numbers that they form 
emboli, which not only close, but actually distend the capil- 
laries and larger bloodvessels, and interfere with the normal 
functions of the organ. A similar condition is sometimes 
found on microscopical examination of the liver, spleen, 
and lungs. From these appearances, it was inferred by 
Bollinger that the bacilli produce the diseased condition 
simply by accumulating in large numbers in these impor- 
tant organs, and mechanically interrupting their functions. 
This is known as the mechanical interference theory. 
Klebs and Toussaint were formerly ardent advocates 
of this theory in its application to anthrax, and the latter 88 
BACTERIAL POISONS. 
thought that the symptoms and death are due to stoppage 
of the pulmonary circulation by means of emboli. How- 
ever, Hoffa studied this point by making numerous post- 
mortem examinations, and was unable to confirm it. A 
like result followed the work of Virchow, Colin, and 
Siedamgrotzky, and the mechanical-interference theory 
has been abandoned. 
In the majority of germ diseases this theory never had 
any support. There is not found any great accumulation 
of bacteria in any organ, and the number and distribution 
of the germs are such that the theory of mechanical inter- 
ference cannot be held. 
3. Another answer given to the question, How do germs 
cause disease? is, that they do so by consuming the proteids 
of the body and thus deprive it of its sustenance. The 
proteids are known to be necessary for the building up of 
eells, and it is also known that microorganisms feed upon 
proteids. But this theory is untenable for several reasons. 
In the first place, many of the infectious diseases destroy 
life so quickly that the fatal effect cannot be supposed to 
be due to the consumption of any very large amount of 
proteids. In the second place, the distribution of the micro- 
organisms is such in many diseases that they do not come 
in contact with any large proportion of the proteids of the 
body. In the third place, the symptoms of the majority of 
these diseases are not those which would be produced by 
withdrawing from the various organs their food. The 
symptoms are not those of general starvation. 
4. Still another theory, which has been offered, is that 
the bacteria destroy the blood corpuscles, or lead to their 
rapid disintegration. But in many of the infectious dis- 
eases, as has been stated, the microorganisms, although very 
abundant in some organs, are not present in the blood. 
Moreover, the disintegration of the blood corpuscles is not 
confirmed by microscopical examination. 
5. Seeing the vital deficiencies in the above theories, and 
being impressed by the results obtained by the chemical 
study of putrefaction, bacteriologists have been led to in- 
quire into the possibility of the symptoms of the infectious RELATION TO INFECTIOUS DISEASES. 
89 
diseases being due to chemical poisons. In investigating 
this theory, three possibilities suggest themselves : 
(a) The microorganisms themselves may be poisonous, 
or the poison may be an integral part of them. Neelsen, 
at one time an advocate of this theory, thus accounted for 
the appearance and increase in violence of the symptoms as 
the germs increase in number. In order for the conditions 
of this theory to be fulfilled the microorganisms must be 
present in the blood before any of the symptoms appear. 
But in anthrax the most thoroughly studied of all the in- 
fectious diseases, and the one to which all these theories 
have been applied, the bacilli first appear in the blood, as a 
rule, only a few hours before death, and long after the 
appearance of the first symptoms; while in many other 
diseases the germs are never found in the blood. More- 
over, as Hoffa has shown, if this theory be true, the in- 
jection of a large quantity of anthrax bacilli directly into 
the blood should be followed immediately by symptoms of 
the disease, and death should be speedy. But lie found, on 
making experiments of this kind, that the symptoms did 
not appear until from twenty-four to seventy-two hours. 
Nencki found by analysis that the substance of the an- 
thrax bacilli resembles vegetable casein in some respects, 
and animal mucin in others. This “ anthrax protein ” is 
freely soluble in alkalies, is insoluble in water, acetic acid, 
and the dilute mineral acids. It contains no sulphur and 
was believed by Nencki to be inert; but the recent re- 
searches of Buchner has shown that this belief is not well 
founded. It has been stated by a number of investigators 
that suppuration might lie induced by the injection of cer- 
tain sterilized cultures, but the dictum of Weigert, “no 
suppuration without bacteria,” has been generally accepted ; 
and statements to the contrary, although some of them have 
been made by men of excellent reputation, have until recently 
received but little credence. However, Buchner has shown 
conclusively that the albuminate of the bacterial cell in as 
many as seventeen different species possesses well-marked 
pyogenetic properties, and that the pus formed is free from 
germs. Buchner separated the microorganisms from the 90 
BACTERIAL POISONS. 
soluble substances accompanying them by sedimentation and 
decantation, washed the cells, dissolved them in a 0.5 per 
cent, solution of potash by the aid of heat, precipitated the 
albumin with dilute mineral acid, and, after repeated re- 
solution in alkali and reprecipitation with acid, employed 
the purified proteid in his experiments. Introduced with 
antiseptic precautions under the skin, this substance invari- 
ably causes suppuration. This demonstrates that the sub- 
stance of the bacterial cell is not altogether inert. It is 
impossible at present to say to what extent the course of 
an infectious disease may be influenced by the breaking 
down of a large number of bacterial cells and the intro- 
duction of their substance into the blood. 
(b) The microorganisms may be intimately associated 
with or may produce a soluble, chemical ferment, which, 
by its action on the body, produces the symptoms of the 
disease and death. This theory formerly had a number of 
ardent supporters, among whom might be mentioned the 
eminent scientist, de Bary. But Pasteur proved the 
theory false when he filtered anthrax blood through earthen 
cylinders, inoculated animals with the filtrate, and failed to 
produce any effect. Hencei made a similar demonstration 
when he inoculated a two per cent, gelatin preparation 
with the anthrax bacillus, which liquefied the preparation, 
and on standing the bacilli settled to the bottom. The 
supernatant fluid, which was clear, alkaline in reaction, and 
contained dissolved “ anthraxprotein,” was filtered and 
injected into animals without producing any effect.1 
It must not be inferred from the above statements that 
bacteria do not produce any ferments. Many of them do 
form both diastatic and peptic ferments, which may retain 
their activity after the bacteria have been destroyed; but 
there is no proof that in any case these ferments have any 
causal relation to the disease. After the diseased process 
has been inaugurated some of these ferments probably play 
1 We now know that if the supernatant fluid used in this experiment 
had been injected in sufficient quantity death would have been produced 
by the soluble chemical poisons. RELATION TO INFECTIOUS DISEASES. 
91 
an important part in the production of morphological 
changes, the nature of which will be indicated when these 
diseases are discussed. 
(c) The germ may produce chemical poisons by splitting 
up preexisting complex compounds in the body. This 
theory finds, in the first place, strong support in the well- 
known fact that many of the putrefactive germs produce 
highly poisonous bodies; and, in the second place, the for- 
mation of chemical poisons will account for the appearance 
of the symptoms of the disease when the microorganisms 
never find their way into the blood. The correctness of 
this theory has been tested by a large number of investi- 
gators, and with the result that its truth has been firmly 
established. It was soon found that pathogenic germs 
grown in meat broth and other culture media elaborate 
chemical poisons which, when injected into the lower animals, 
induce in an acute form one or more of the symptoms char- 
acteristic of the disease caused in man by the microorgan- 
ism. It is true that until quite recently this theory has 
been opposed by some, and it is altogether possible that 
at present there may be those who are not altogether con- 
vinced of its truth. However, we are not acquainted with 
any argument against it which remains unanswered. For 
a while Baumgarten claimed that the formation of chem- 
ical poisons in the dead matter of meat broth and other 
media by the germ does not prove that the same agent is 
capable of forming the same or similar products within the 
living body; but the isolation of tetanine from the ampu- 
tated arm of a man with tetanus, by Brieger, furnished 
the first positive answer to this criticism, and since that 
time a number of bacterial poisons have been obtained from 
the bodies of men and the lower animals. We now expect 
to find each specific, pathogenic microorganism producing 
its characteristic poison or poisons. The evidence on 
this point will be given further on in a brief sketch of the 
chemical factors in the causation of some of the best-known 
infectious diseases. 
Before taking up the individual diseases, we will give 92 
BACTERIAL POISONS. 
what appears to us, in the present state of* our knowledge, 
a correct definition of an infectious disease. 
An infectious disease arises when a specific, pathogenic 
microorganism, having gained admittance to the body, and 
having found the conditions favorable, grows and multi- 
plies, and in so doing elaborates a chemical poison which 
induces its characteristic effects. 
In the systemic infectious diseases, such as anthrax, 
typhoid fever, and cholera, this poison is undoubtedly taken 
into the general circulation, and affects the central nervous 
system. In the local infectious diseases, such as gonorrhoea, 
and infectious ophthalmia, the principal action of the poison 
seems to be confined to the place of its formation. Though 
even in these, when of a specially virulent type, the effects 
may extend to the general health. It may be that in some 
diseases the chemical poison has both a local and a systemic 
effect. Thus, it is by no means certain that the ulceration 
of typhoid fever is due directly to the bacillus. On the 
other hand, it is altogether probable that the anatomical 
changes in the intestine result from the irritating effects of 
the poison at the place of its formation. 
With the proof, that the deleterious effects wrought by 
germs are due to chemical poisons elaborated by them 
during their growth, admitted, let us inquire what proper- 
ties a microorganism must possess before it can be said to 
be the specific anise of a 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: 
1. The germ must present in all cases of that disease. 
2. The germ must be isolated from other organisms 
and from all other matter found with it in the diseased 
animal. 
3. The germ thus freed from all foreign matter must, 
when properly introduced, produce the disease in healthy 
animals. 
4. The microorganism must be found properly dis- RELATION TO INFECTIOUS DISEASES. 
93 
tributed in the animal in which the disease lias been 
induced. 
Let us give our special attention to the first of these 
rules for a few moments. What is meant by the state- 
ment 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 disease in America, decide whether 
or not a bacillus which he finds is identical with one which 
lias been reported as invariably present in the same disease 
by B, who has investigated an epidemic in Germany? 
W hat 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 staining reagents, 
manner of growth on various nutrient media, and, in ex- 
ceptional 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 property by which the germ causes the disease 
is left wholly out of consideration. It is admitted that any 
causal relation which the germ may have to the disease is 
due to its capability of forming one or more chemical poi- 
sons, and yet no attempt is made to ascertain whether or 
not it possesses this property. Indeed, some of the most 
eminent bacteriologists have taught that in the identifica- 
tion of germs the reactions with staining reagents and the 
appearance of the growths on the various nutritive media 
are of more importance than the observation of the effects 
upon animals. Thus, Flugge 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 inoculation or feeding have 
been made with impure material containing other active 
bacteria. It is known that a group of widely distributed 
organisms, which, however, are wholly different from the 
typhoid bacillus, have the power, when injected subcu- 
taneously or intravenously, of producing in animals death 
with marked swelling and ulceration of Peyer’s patches. 
To these organisms undoubtedly are due the apparently 94 
BACTERIAL POISONS. 
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 germ found in the spleen of man after death from 
typhoid fever, because they do not react in the same man- 
ner with the anil in stains, and present a different appear- 
ance in their growths on potatoes. 
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 inocu- 
lates animals, and these manifest all the signs, together with 
the appearance of the characteristic membrane of diphtheria, 
and in these animals lie finds his bacillus growing as in the 
throats of the children. All the rules of Koch have been 
complied with. Has A demonstrated that his bacillus is the 
sole cause of diphtheria ? No. 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 diphtheria. 
The most which can be proven by Koch’s rules is that a 
given germ is a cause of a certain disease. They do not 
show, as most bacteriologists would have us believe, that 
the given 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, produces certain effects which are due to the alka- 
loid, caffein, and which invariably follow the drinking of 
a decoction of these berries; would our supposed discoverer 
be justified in concluding that the coffee tree is the only 
plant 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 of China and the guarana of South America. 
The moment that it is granted that the real poison of the RELATION TO INFECTIOUS DISEASES. 
95 
disease is chemical in character, it becomes evident that no 
one is justified in saying that one germ is the sole source of 
that poison. Such a statement would be as unwarranted 
as one that the coffee tree is the sole source of caffein, or 
that the stryclmos Ignatii is the only species of the nat- 
ural order Logan iacese 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, 
but by the character of the chemical poison which is the true 
materies morbi. 
Bacteria cannot be classified, so far as their causal rela- 
tionship to disease is concerned (and this is the most im- 
portant knowledge to be gained from them), until we know 
the nature of their chemical products, for it is by virtue of 
these that the germs have any causal relationship to dis- 
ease. 
It is possible that two germs maybe 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. This is 
not only a possibility, it is a fact which has been demon- 
strated repeatedly, both with pathogenic and non-pathogenic 
organisms. A few illustrations may be given here: The 
veast plant is not the only microorganism which will pro- 
duce alcohol in saccharine solutions. The same product 
results from the growth of the bacterium Bischleri, bac- 
terium coli commune, bacterium ilei, bacterium ovale ilei, 
bacterium lactis aerogenes, and others (Nencki). Mor- 
phologically, there are marked differences between the yeast 
plant and these bacteria, but they alike produce alcohol. 
More than a dozen germs, including both micrococci and 
bacilli, are capable of generating lactic acid. Some of these 
produce an acid which is optically inactive; others, one 
which is dextro-rotatory; and others still, one which is lsevo- 
rotatory. The tetanus germ of Kitasato and that of 
Tizzoni and Cantani are known to be different. Cultures 
of the former in bouillon are virulent, while those of the 96 
bacterial poisons. 
latter in the same medium are inert. Not only are these 
two organisms morphologically and biologically distinct, 
but their poisons are chemically unlike. Brieger and 
Franker precipitated the poisonous albumin of the germ 
of Kitasato with alcohol, but this reagent renders the 
poison of the Italian germ inert. Notwithstanding this 
difference, however, both microorganisms and their chemical 
products produce tetanic convulsions and death in the lower 
animals. We 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. All attempts to find a mor- 
phologically specific germ in the summer diarrhoeas of 
infancy have failed. The labors of Booker in this coun- 
try and of Escherich in Germany have shown that no 
one species or variety is constantly present. No less than 
thirty distinct germs have been obtained from the bowels 
and feces of children suffering from these diarrhoeas. A 
germ which is frequently present one season may not be 
found at all the next. Are we to conclude from this fail- 
ure to comply with the first of Koch’s rules, that the sum- 
mer diarrhoeas of infancy are not due to microorganisms? 
Certainly not; especially in view of the fact that Baginsky 
and Stadthagen have obtained from pure cultures of a 
saprophytic germ found in the stools of cholera infantum a 
poisonous base and a poisonous proteid; and Vaughan 
has shown that at least three of Booker’s bacteria pro- 
duce chemical poisons which cause in kittens retching, 
vomiting, purging, collapse, and death. To the contrary 
we are justified in concluding that these diarrhoeas may be 
due to any one or more of a number of germs which differ 
from one another sufficiently morphologically 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 agents, but by a study of their 
chemical products, the agents by virtue of which they cause 
the disease. 
We think that we are justified in concluding that in 
those diseases in which the four rules of Koch have been RELATION TO INFECTIOUS DISEASES. 
97 
complied with, the germ is a cause of the disease, but our 
range of observation must be much wider than it now is 
before we can say that the given germ is the only cause of 
the disease. 
We believe that those few infectious diseases, such as 
anthrax and tuberculosis, which have such well-marked, 
typical, clinical histories, are due to equally well-marked 
and morphologically distinct microorganisms which can he 
recognized by microscopical study alone; but we do not 
believe that this is true in diseases showing such wide vari- 
ations in symptoms as is the case in typhoid fever and 
cholera infantum. 
In all cases, we insist that the true test of the specific 
character of a germ is to be made with its chemical pro- 
ducts. A given bacterium may not multiply in the circu- 
lating blood of a dog, and failure to do so is by no means 
proof that the same organism might not cause disease in 
man; but every germ which causes disease in man does so 
by virtue of its chemical products, and if these be isolated 
and injected into the dog in sufficient quantity a poisonous 
effect will be produced. In the study of the bacteriology 
of the infectious diseases, the third and fourth of Koch’s 
rules have not been complied with in many diseases on 
account of the insusceptibility of the lower animals. The 
majority of investigators, meeting with this difficulty, have 
been inclined to rest content with the first two rules, and 
to conclude that when a given germ is constantly present 
in a given disease, and not found in other diseases, that it 
is the cause of the disease with which it is associated. In- 
deed, we find so good an authority as Welch stating that 
the successful inoculation of animals is not necessary in 
order to prove the causal relationship of a germ to a disease. 
In 1889, Vaughan suggested that in those instances in 
which the third and fourth of Koch’s rules cannot be 
complied with on account of the insusceptibility of the 
lower animals, it must be shown that the germ can pro- 
duce chemical poisons which will induce in the lower ani- 
mals in an acute form the characteristic symptoms of the 98 
BACTERIAL POISONS. 
disease, before the proof that the given germ is the cause 
of the disease be accepted as positive. 
Heretofore, the science of bacteriology has been largely 
founded upon morphological studies. Bacteriologists have 
given their time and attention to the discovery of bacterial 
forms in the diseased organism and to observations of char- 
acteristics in structure and growth of different species of 
bacterial life. We must now study the physiology and 
chemistry of the 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 yeast plant 
was limited toils form and method of growth ; of how little 
practical importance this knowledge would be. That the 
yeast plant requires a saccharine soil before it can grow, 
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 condi- 
tions under which pathogenic germs multiply and the pro- 
ducts which they elaborate in their multiplification 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, 
we wish that it may be plainly understood that we are not 
offering any hostile criticism upon the great men who have 
done this work and who have formulated conclusions there- 
from. The development of bacteriology has been in accord- 
ance with the natural law governing the growth of all the 
biological sciences. The study of form naturally and neces- 
sarily 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 varia- 
tions more or less marked are likely to be found in other 
individuals. More time and wider opportunities of ob- RELATION TO INFECTIOUS DISEASES. 
99 
servation 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. 
We repeat that it is no discredit to the science nor to the 
men who have developed it to say that the study of bac- 
teriology has hitherto been almost wholly morphological. 
Without the morphologist the physiologist and the physio- 
logical chemist could 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 fundamental lines of work must be altered in 
order to correspond with this new knowledge. 
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 toxi- 
cogenic. This would necessitate a chemical study of all kinds 
of bacteria, both the pathogenic and the non-pathogenie. 
Every fact ascertained 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. 
Secondly, it must be determined under what conditions 
these germs are toxieogenic. 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 100 
BACTERIAL POISONS. 
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 con- 
ditions 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 treatment which is 
efficient in 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 
infectious 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 chief energy upon the central 
nervous system, 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. CHAPTER V. 
THE BACTERIAL POISONS OF SOME OF THE INFECTIOUS 
DISEASES. 
We will now give our attention to the chemical poisons, 
both the ptomaines and the proteids, of some of the infec- 
tious diseases, and in doing this we will illustrate and sub- 
stantiate the statements made in the preceding chapter. 
Anthrax.—Tlie definition of an infections disease, as 
we have given it, is well illustrated by the facts which have 
been learned concerning the causation of anthrax, which 
has probably been more thoroughly studied than any other 
infectious disease. Kauscii taught that this disease has its 
origin in paralysis of the nerves of respiration. As to the 
cause of this paralysis he gave us no information. Delafond 
thought that anthrax has its origin in the influence of the 
chemical composition of the soil affecting the food of ani- 
mals and leading to abnormal nutrition. The investigations 
of Gerlach in 1845 demonstrated the contagious nature 
of the disease, which was emphasized by Heusinger in 
1850 and accepted by Virchow in 1855. However, as 
early as 1849, Poleender found numerous rod-like micro- 
organisms in the blood of animals with the disease. This 
observation was confirmed by Brauell, who produced 
the disease in healthy animals by inoculations with matter 
taken from a pustule on a sick horse. Attempts were made 
to ridicule the idea that these germs might be the cause of 
the disease, and it was said that the bodies seen were only 
fine shreds of fibrin or blood crystals. Some claimed that 
the rod-like organisms reported were due to defects in the 
glass, while others claimed that the defects existed in the 
eye of the observer, and others still suggested that the de- 102 
BACTERIAL FOISONS. 
fects might be found back of the eye and in the brain. But 
in 1863, Davaine showed that these little bodies must 
have some causal relation to the disease, inasmuch as his 
experiments proved that inoculation of healthy animals 
with the blood of those sick with anthrax produced the 
disease only when taken at a time when the blood con- 
tained these organisms. He also demonstrated beyond any 
question that these rod-like bodies are bacteria, capable of 
growth and multiplication. The conclusions of this investi- 
gator were combated by many; but Pasteur, Koch, 
Bollinger, de Barry, and others, studied the morph- 
ology and life-history of these organisms, and then came 
the brilliant results of Pasteur and Koch in securing 
protection against inoculation anthrax by the vaccination 
of healthy animals with the modified germ and subsequent 
inoculation with the virulent form. Now, the bacillus 
anthracis is known in every bacteriological laboratory, and 
by inoculation with it the disease is communicated at will 
to susceptible animals. But here the question arose, How 
do these bacilli produce anthrax? and in answer to this 
question the various theories which we have mentioned 
were proposed. 
The first successful attempt to study the chemical poisons 
of anthrax was made by IIoffa, who obtained from pure 
cultures of the bacillus small quantities of a ptomaine, 
which, when injected under the skin of animals, produces 
the symptoms of the disease and death. This substance 
causes at first increased respiration and action of the heart, 
then the respirations become deep, slow, and irregular; 
the temperature falls below the normal ; the pupils are 
dilated, and a bloody diarrhoea sets in. On section the 
heart is found contracted, the blood dark, and ecchymoses 
arc observed on the pericardium and peritoneum. Hoffa 
names his poison anthracin. Recently Hoffa has isolated 
this poison from the bodies of animals dead from anthrax. 
It has been said that Hoffa’s work was the first suc- 
cessful attempt to study the chemical poisons of anthrax. 
However, his results cannot be considered altogether satis- 
factory. The small amount of the basic substance which ANTHRAX, 
103 
he obtained rendered it highly probable that in the ease of 
a germ .so virulent as that of anthrax there must be other 
chemical poisons produced. This supposition has been con- 
firmed by the labors of Hankin, who, in 1889, while at 
work in Koch’s laboratory, prepart'd from cultures of 
the bacillus anthracis an albumose which, when employed 
in comparatively large amount, proved fatal to animals, 
but when used in very small quantity gave immunity 
against subsequent inoculations with the living germ. 
Unfortunately, Han kin does not mention the symptoms 
induced by toxical doses of this substance. Whether or 
not the albumose of Hankin contains in statu nascendi 
the base of Hoff a, and owes its poisonous properties to 
the same, has not been determined. 
Brieger and Frankel obtained the poisonous proteid 
of anthrax from animals in which the disease had been 
induced by inoculation with the bacillus. The liver, spleen, 
lungs, and kidneys of these animals were finely divided 
and rubbed up with water. After this had stood in a 
refrigerator for twelve hours it was passed through a 
Chamberland filter and the proteid precipitated from the 
filtrate with absolute alcohol. 
Martin, by growing the anthrax bacillus for from ten 
to fifteen days in an alkaline albuminate from blood serum 
and filtration through porcelain, obtains the following 
metabolic products: 
1. Protoalbumose and deuteroalbumose and a trace of 
peptone. All of these react chemically like similar sub- 
stances prepared by peptic digestion. 
2. An alkaloid. 
3. Small quantities of leucin and tyrosin. 
The most characteristic property of the albumoses is that 
their solutions are strongly alkaline, and the alkalinity is 
not removed by treatment with alcohol, benzol, chloroform, 
or ether, or by dialysis. 
The alkaloid is soluble in water, alcohol, and amylic 
alcohol; insoluble in chloroform, ether, and benzol. Its 
solutions are strongly alkaline and the alkaloid forms crys- 
talline salts with acids. It is precipitated by the general 104 
BACTERIAL POISONS. 
alkaloidal reagents, with the exception of potassio-mercuric 
iodide. It is somewhat volatile and loses its poisonous 
properties on exposure to the air. 
The mixed albumoses are poisonous only in considerable 
doses, 0.3 gramme being required to kill a mouse of 22 
grammes weight when injected subcutaneously. Smaller 
doses cause a local oedema and a somnolent condition, from 
which the animal recovers. The larger doses produce a more 
extensive oedema and the somnolence deepens into coma, 
terminating in death. In some cases the spleen is enlarged. 
The absence of germs was demonstrated by plate cultures. 
The alkaloid causes similar symptoms. It is, however, 
more poisonous and acts more rapidly than the albumoses. 
The animal is affected immediately after the injection, and 
the gradually increasing coma terminates in death. The 
alkaloid also produces oedema, and in many cases throm- 
bosis of the small veins. Extravasation into the peritoneal 
cavity is occasionally seen, and the spleen is ordinarily 
enlarged and filled with blood. The fatal dose for a mouse 
is from 0.1 to 0.15 gramme, death resulting within three 
hours. 
This alkaloid does not appear to be identical in its action 
with the anthracin of Hoffa. 
Asiatic Cholera.—There are good reasons, apart from 
experimental evidence, for believing that the comma bacillus 
of Kocii produces its ill effects by the elaboration of chemi- 
cal poisons. This germ is not a blood parasite. It grows 
only in the intestine, and the symptoms of the disease and 
death must result from the absorption of its poisonous 
products. In confirmation of this statement experiment 
has shown that this is one of the most active, chemically, 
of all known pathogenic germs. 
In the first place, Bitter has shown that the comma 
bacillus produces in meat-peptone cultures a peptonizing 
ferment, which remains active after the organism has been 
destroyed. Like similar chemical ferments, it converts an 
indefinite amount of coagulated albumin into peptone. It 
is more active in alkaline than in acid solutions, thus 105 
ASIATIC CHOLERA. 
resembling pancreatin more than pepsin. This resem- 
blance to pancreatin is further demonstrated by the fact 
that its activity is increased by the presence of certain 
chemicals, such as sodium carbonate and sodium salicylate. 
That a diastatic ferment is also produced by the growth of 
the bacillus was indicated in the experiments of Bitter by 
the development of an acid in nutrient solutions contain- 
ing starch paste. However, all attempts to isolate the 
diastatic ferment were unsuccessful. A temperature of 60° 
destroys or greatly decreases the activity of ptyalin, and 
this seems to be true also of the diastatic ferment produced 
by the comma bacillus. But the formation of an acid from 
the starch pre-supposes that the starch is first converted into 
a soluble form. 
Fermi has succeeded in isolating the peptonizing ferment 
of the cholera germ in the following manner : 65 per cent, 
alcohol added to gelatin which has been liquefied by the 
bacillus precipitates the proteid, but not the ferment. After 
twenty-four hours the precipitate is removed by filtration 
and the ferment precipitated from the filtrate by the addi- 
tion of absolute alcohol. After being collected on a filter 
and dried the ferment is dissolved in an aqueous solution 
of thymol and its peptonizing properties demonstrated on 
gelatin tubes. 
Rietsch believes that the destructive changes observed 
in the intestines in cholera are due to the action of the 
peptonizing ferment. 
Cantani injected sterilized cultures of the comma bacil- 
lus into the peritoneal cavities of small dogs and observed 
after from one-quarter to one-half hour the following symp- 
toms : Great weakness, tremor of the muscles, drooping of 
the head, prostration, convulsive contractions of the pos- 
terior extremities, repeated vomiting, and cold head and ex- 
tremities. After two hours these symptoms began to abate, 
and after twenty-four hours recovery seemed complete. 
Control experiments with the same amounts of uninfected 
beef-tea were made with negative results. The cultures 
used were three days old when sterilized. Older cultures 
seemed less poisonous and a high or prolonged heat in 106 
BACTERIAL poisons. 
sterilization decreased the toxicity of the fluid. From these 
facts Cantani concluded that the poisonous principle is 
volatile, but the effect of high or prolonged heat in dimin- 
ishing the toxicity was more probably due to its destructive 
effect on the poisonous proteids. 
Cantani also observed that the blood of those sick with 
cholera is acid : this has been confirmed by Strauss by 
the examination of the blood directly after death; and 
Ahrend found lactic acid in the strongly acid urine of a 
cholera patient. 
Nicati and Rietsch produced fatal effects in dogs by 
injecting cultures, from which all germs had been removed 
by filtration, into the bloodvessels. Later, the same inves- 
tigators obtained from old bouillon cultures containing 
peptone a poisonous base. Ermengen also showed that 
cultures after filtration through a Chamberland filter are 
poisonous. 
Klebs has attemped to answer experimentally the ques- 
tion, In what way does the cholera germ prove harmful ? 
Cultures of the bacillus in fish preparations were acidified, 
filtered, the filtrate evaporated on the water-bath, the residue 
taken up with alcohol and precipitated with platinum chlo- 
ride. The platinum was removed with hydrogen sulphide, 
and the crystalline residue obtained on evaporation was 
dissolved in water and injected intravenously into rabbits. 
Muscular contractions were induced. Death followed in 
one animal, which, in addition to the above treatment, 
received an injection of a non-sterilized culture. In this 
ease there was observed an extensive calcification of the 
epithelium of the uriniferous tubules. Klebs believes this 
change in the kidney to be induced by the chemical poison, 
and from this standpoint he explains the symptoms of 
cholera as follows : The cyanosis is a consequence of arte- 
rial contraction, the first effect of the poison. The mus- 
cular contractions also result from the action of the poison. 
The serous exudate into the intestines follows upon epithe- 
lial necrosis. Anuria and the subsequent symptoms appear 
when the formation and absorption of the poison become 
greatest. ASIATIC CHOLERA. 
107 
Hueppe states that the severe symptoms of cholera can 
be explained only on the supposition that the bacilli produce 
a chemical poison, and that this poison resembles muscarine 
in its action. 
Vi leiers isolated by the Stas-Otto method from two 
bodies dead from cholera, a poisonous base which was liquid, 
pungent to the taste, and possessed the odor of hawthorn. 
It was strongly alkaline, and gave precipitates with the 
general alkaloidal reagents. From one to two milligrammes 
of this substance, injected into frogs, caused decreased 
activity of the heart, violent trembling, and death. The 
heart was found in diastole, and full of blood, and the 
brain slightly congested. However, the presence of this 
substance in the bodies of persons who have died of cholera 
does not prove that its production is due to the cholera 
bacillus. 
Poucitet extracted from cholera stools, with chloroform, 
an oily base belonging to the pyridine series. It readily 
reduces ferric as well as gold and platinum salts, and forms 
an easily decomposable hydrochloride. It is a violent poison, 
irritating the stomach, and retarding the action of the heart. 
Subsequently, he obtained an apparently identical substance 
from cultures of Koch’s comma bacillus. 
In 1887, Brieger made a report of his studies on the 
chemistry of the cholera bacillus. He used pure cultures on 
beef-broth (fleischbrei), which was rendered alkaline by the 
addition of a 3 per cent, soda solution. These were kept 
at from 37° to 38°. After twenty-four hours, cadaverine 
was found to be present. Older cultures furnished very 
small quantities of putrescine, but cultures on blood-serum 
yielded much larger amounts of this base. While eada- 
verine and putrescine cannot be said to be poisonous, they 
do cause necrosis of tissue into which they are injected, and 
their formation by the cholera bacillus may account for the 
necrotic tissue in the intestine in the disease. The lecithin 
of the beef-broth was slowly acted upon by the germs, but 
with age the amount of choline increased, reaching its 
maximum during the fourth week. 
Creatine proved still more resistant to the action of the 108 
BACTER1AL POISONS. 
germs; but, after six weeks, a considerable quantity of 
creatinine was isolated, and a smaller amount of methyl- 
guanidine. The latter is very poisonous, causing muscular 
tremors and dyspnoea. The presence of methyl-guanidine 
indicates that the comma bacillus acts as an oxidizing agent, 
since creatine yields methyl-guanidine only by oxidation. 
Brieger succeeded in finding, in addition to the above- 
mentioned ptomaines, which are common products of putre- 
faction, two poisons which he considers as specific products 
of the comma bacillus. One of these, found in the mer- 
curic chloride precipitate, is a diamine, resembling trime- 
thylenediamine. It produced muscular tremor and heavy 
cramps. In the mercury filtrate was found another poison, 
which, in mice, produced a lethargic condition ; the respira- 
tion and heart’s action became slow, and the temperature 
sank, so that the animal felt cold. Sometimes there was 
bloody diarrhoea. 
BRiEGERand Frankel found that the insoluble proteid 
which they obtained from cultures of the cholera bacillus, 
when suspended in water and injected subcutaneously in 
guinea-pigs, caused death after from two to three days. 
Section showed inflammatory swelling and redness of the 
subcutaneous tissue, extending into the muscles for some 
distance about the point of injection, but no necrosis. 
There was no change in the intestines and no effusion into 
the peritoneum. In some instances there were evidences 
of beginning fatty degeneration of the liver. Upon rab- 
bits this substance, even in large doses, was without effect. 
In endeavoring to obtain immunity in guinea-pigs against 
cholera, Gamaleia employs cultures which have been ster- 
ilized at 120°. Subcutaneous injections of these cause 
transient oedema, and the animals soon recover. The high 
temperature destroys not only the bacillus, but renders inert 
certain “ ferment-like ” products. However, if the cultures 
be sterilized at 60°, large doses (10 c.e. per kilogramme, body 
weight) cause death, injected intravenously in rabbits, and 
a less amount produces marked symptoms. The animals 
refuse food, and a diarrhoea, which may continue for hours, 
appears. Often there is cloudiness of the cornea and reten- ASIATIC CHOLERA. 
109 
tion of urine, which is albuminous. The animals recover 
very slowly. In this connection Bouciiard remarks that 
in 1884 he obtained by the intravenous injection of the 
urine of a cholera patient in rabbits muscular tremor, cyan- 
osis, albuminuria, and diarrhoea, but that he has never suc- 
ceeded in inducing these symptoms with the cholera vibrio. 
Petri finds that the comma bacillus produces in solu- 
tions of peptone large amounts of tyrosin and leucin, a 
small quantity of indol, fatty acids, poisonous bases, and a 
poisonous proteid. The proteid resembles peptone in its 
behavior toward heat and chemical reagents, and is desig- 
nated by Petri as “ toxopeptone.” It is not precipitated 
by heat or concentrated nitric acid, nor by potassium ferro- 
cyanide and acetic acid, nor by ammonium sulphate added 
to saturation. With sodium phospho-tungstate it gives a 
precipitate which clears up on the application of heat. The 
precipitate with tannic acid is insoluble in an excess of the 
precipitant. It gives the biuret reaction perfectly, but 
responds to Millon’s test but feebly. 
In quantities of 0.30 of a gramme per kilogramme and 
more it is fatal to guinea-pigs within eighteen hours. It 
produces muscular tremor and paralysis. Post-mortem 
shows an effusion into the peritoneal cavity, marked injec- 
tion of the bloodvessels of the intestines, and isolated 
hemorrhagic spots. 
This proteid is not rendered inert by a temperature of 
100°. Petri does not claim that he has obtained a chemi- 
cally pure body, but supposes that it is contaminated with 
more or less unchanged peptone. 
Scholl has studied the chemical products of the cholera 
bacillus when grown under anaerobic conditions. Fresh 
eggs were sterilized and inoculated in the usual way. The 
eggs, after being kept for eighteen days at 36°, were opened. 
The contents smelled intensely of hydrogen sulphide, but 
not of amines. The albumin was completely fluid, while 
the yolk was more solid and of a dark color. 
Five c c. of the fluid contents were injected into the 
abdomen of a guinea-pig. Soon the posterior extremi- 
ties were paralyzed, and after ten minutes the paralysis 110 
BACTERIAL POISONS. 
became general, the animal lying on the side. After five 
minutes more convulsive movements of the extremities 
began, and forty minutes after the injection the animal was 
dead. Section showed the vessels of the small intestines 
and stomach highly injected, a colorless effusion in the 
peritoneal cavity, and the heart in diastole. 
The albuminous content of the egg was poured into ten 
times its volume of absolute alcohol. The precipitate was 
collected and washed with alcohol until a colorless filtrate 
was obtained. The precipitate was then digested for fif- 
teen minutes with 200 c.c. of water and filtered. Eight 
c.c. of the filtrate was injected into the abdomen of a 
guinea-pig. Paralysis resulted immediately, and within 
one and one-fourth minutes the animal was dead. Section 
showed marked injection of the vessels of the small intes- 
tines, a bloody transudate in the peritoneal cavity and the 
heart in diastole. 
The poisonous proteid was rendered inert by a tempera- 
ture of 100°; it was not altered by short exposure to 75°, 
but attempts to evaporate the solution at 40° in vacuo over 
calcium chloride destroyed the poisonous properties. The 
proteid was finally precipitated from its aqueous solution 
by a mixture of alcohol and ether. It was washed with 
ether and the ether allowed to evaporate spontaneously. A 
small bit of this proteid proved fatal to guinea-pigs, and 
the same post-mortem changes were found as given above. 
Scholl classes this proteid among the peptones. It is not 
precipitated by heat or concentrated nitric acid, singly or 
combined, nor by ammonium sulphate. It gives the xantho- 
proteid and biuret reactions. Scholl regards this as the 
true poison of cholera, and points out its difference from 
the proteid of Brieger and Frankel and that of 
Petri. 
Bujwid found that on the addition of from five to ten 
per cent, of hydrochloric acid to bouillon cultures of the 
cholera bacillus there was developed after a few minutes a 
rose-violet coloration which increased during the next half 
hour and in a bright light showed a brownish shade. The 
coloration is more marked if the culture is kept at about 111 
ASIATIC CHOLERA. 
37°. In impure cultures this reaction does not occur. 
The Finkler-Prior bacillus cultures give after a longer 
time a similar, but more of a brownish coloration. Cul- 
tures of many other bacilli were tried and failed to give 
this reaction.1 
Brieger found that this color is due to an indol deriva- 
tive. In cholera cultures on albumins he obtained indol 
by distillation with acetic acid. 
Bujwid has made a further contribution to our knowl- 
edge of the “cholera-reaction.” His conclusions are as 
© 
follows: 
(1) Five to ten per cent, of hydrochloric acid added to 
cholera cultures produce a rose-violet coloration, which is 
characteristic of the comma bacillus. 
(2) No other bacterium gives the same coloration under 
the same conditions. 
(3) The coloration appears in such cultures which are 
from ten to twelve hours old, so that this test can be used 
for diagnostic purposes, and will give results before they 
can be obtained by plate cultures. 
(4) Impure cultures do not give this reaction. 
Dunham finds the best medium for the “ cholera-reac- 
tion” to be a one per cent, alkaline peptone solution with 
one-half per cent, of common salt. Bujwid prefers a two 
per cent, feebly alkaline peptone solution with salt. Jadas- 
sohn finds that gelatin cultures give the reaction both 
before and after the liquefaction of the gelatin. The un- 
dissolved gelatin, after the addition of hydrochloric or 
sulphuric acid, becomes rose-violet. 
Cohen claims that cultures of other bacilli give a similar 
coloration, but Bujwid explains that the results obtained 
by Cohen were due to the use of impure acids, which con- 
tained nitrous acid. Salkowski agrees with Bujwid, and 
states that, when acids wholly free from nitrous acid are 
used, the reaction is characteristic of the comma bacillus. 
He explains the reaction by supposing that the germ pro- 
1 Poehl deserves the credit of being the first to call attention to this 
reaction, though his work was evidently unknown to Bujwid at the time 
when the latter published his report. 112 
BACTERIAL POISONS. 
duces nitrous acid, which exists in the culture as a nitrite. 
On the addition of' an acid the nitrous acid is set free, and 
acting upon the iridol, which is also present, gives the 
coloration. 
From a very exhaustive research on the importance of 
this test Petri eoines to the following conclusions : 
(1) Seven pure cultures of the cholera germ from as many 
sources gave the reaction with equal distinctness. 
(2) Of one hundred other bacteria tested in the same 
way twenty gave a red coloration. In nineteen of these 
the coloration is due to the nitroso-indol reaction of 
Baeyer. The twentieth (anthrax) gave a color which is not 
due to indol. 
(3) In case of the cholera germ and the others as well, 
the reaction is due to the reducing action of the bacteria on 
nitrates. The reaction is most marked at blood-tempera- 
ture and with the cholera bacillus ; it is least distinct with 
the bacilli of Finkler and Miller. 
(4) None of these bacteria convert ammonia into nitrite. 
(5) The simple addition of sulphuric acid is sufficient to 
give the test, which, however, is most marked when the 
nutritive solution contains 0.01 per cent, of nitrate. 
(6) The reaction is most marked if the sulphuric acid be 
added after the addition of a very dilute nitrite solution. 
Schuchardt calls attention to the fact that Virchow 
observed a red coloration on the addition of nitric acid to 
filtered cholera stools in 1846. Griesinger, in 1885, 
also made mention of the production of a red coloration in 
rice-water stools on the addition of nitric acid. 
A “cholera-blue” has also been observed by Brieger 
in cultures in meat extract containing peptone and gelatin. 
This substance, which is yellow by reflected, and blue by 
transmitted light, is developed by the addition of eoneen- 
trated sulphuric acid to the culture. It may be separated 
from the “cholera-red” as follows : Treat the culture with 
sulphuric acid, then render alkaline with sodium hydrate, 
and extract with ether. Evaporate the ether, and remove 
the “cholera-red” with benzol, then again dissolve the 
“ cholera-blue ” in ether. The characteristic absorption TETANUS. 
113 
bands for this coloring matter begin in the first third of the 
spectrum, between E and F, and darken all of the zone 
lying beyond. 
Winter and Lesage treat a bouillon culture of the 
cholera germ with sulphuric acid, dissolve the precipitate 
in an alkaline medium, reprecipitate with acid, and redis- 
solve in ether, which on evaporation leaves oily drops, 
which, on cooling, form a yellow mass of the appearance 
of a fat This substance is insoluble in water and acids, 
soluble in alkalies and ether. It melts at 50°, and does 
not lose its virulence on being boiled with alcohol rendered 
feebly alkaline. The virulence of a culture and the amount 
of this substance contained therein are in direct proportion 
to each other. 
Small doses of this substance 11 milligramme to 100 
grammes of body weight of the animal) in feebly alkaline 
solution introduced into the stomachs of guinea-pigs cause, 
as a rule, within from four to six hours, a chill, and death 
after twenty-four hours. With larger doses the tempera- 
ture falls after from one-half to one hour, and death results 
within from twelve to twenty hours. Smaller doses cause 
a less marked reaction and the animal recovers within 
twenty-four hours. If killed within this time the animal 
shows a choleraic condition. Rabbits succumb only after 
repeated subcutaneous injections. The substance can be 
extracted from the muscles, liver, kidneys, and urine of 
the poisoned animals. It can also be obtained from cultures 
of a cholera infantum germ. The fact that this poison be- 
longs neither to the ptomaines nor albumins is of interest. 
Cunningham describes ten species of the common ba- 
cillus, one of which does not liquefy gelatin, and fails to 
respond to the cholera reaction. He also states that there 
are cases of undoubted cholera in Calcutta in which the 
common bacillus is wholly wanting. 
Tetanus.—In 1884, Nicolaier, by inoculating 140 
animals with earth taken from different places, produced 
symptoms of tetanus in 69 of them. In the pus which 
formed at the point of inoculation he found micrococci and 114 
bacterial poisons. 
bacilli. Among the latter was one which was someAvhat 
longer and slightly thicker than the bacillus of mouse- 
septicaemia. In the subcutaneous cellular tissue he found 
this bacillus alone, but could not detect it in the blood, 
muscles, or nerves. Heating the soil for an hour rendered 
the inoculations with it harmless. In cultures, Nicolaier 
was unable to separate this bacillus from other germs, but 
inoculations with mixed cultures produced tetanus. In the 
same year, Carle and Ratone induced tetanus in lower 
animals by inoculations with matter taken from a pustule 
on a man just dead from tetanus. In 1886, Rosenbach 
made successful inoculations on animals with matter taken 
from a man who had died from tetanus consequent upon 
gangrene from frozen feet. With bits of skin taken from 
near the line of demarcation, he inoculated two guinea-pigs 
on the thigh ; tetanic symptoms set in within twelve hours, 
and one animal died within eighteen, and the other within 
twenty-four hours. The symptoms corresponded exactly 
with those observed in the “ earth tetanus” of Nicolaier, 
and the same bacillus was found. With mixed cultures of 
this, Rosenbach was also able to cause death by tetanus 
in animals. Beumer had under observation a man who 
died from lockjaw following the sticking of a splinter of 
wood under his finger-nail. Inoculations of mice and 
rabbits with some of the dirt found on the wood led to 
tetanus. The same observer saw a boy die from this dis- 
ease following an injury to the foot from a sharp piece of 
stone. White mice inoculated with matter from the wound, 
and those inoculated with dirt taken from the boy’s play- 
ground, died of tetanus. The bacillus of Nicolaier was 
again detected. Giordano reports the case of a man who 
fell and sustained a complicated fracture of the arm. He 
remained on the ground for some hours, and when assist- 
ance came the muscles and skin were found torn and the 
wounds filial with dirt. On the fifth day he showed symp- 
toms of tetanus, from which he dial on the eighth day. 
Inoculations and examinations for the bacillus were again 
successful. Ferrari also made successful inoculations 
with the blood taken during life from a woman with TETANUS. 
115 
tetanus after an ovariotomy. Hocksinger has confirmed 
the above-mentioned observations by carefully conducted 
experiments, the material for which was furnished by a case 
of tetanus arising from a very slight injury to the hand, 
the wound being filled with dirt. Shakespeare has suc- 
ceeded in inducing tetanus in rabbits by inoculating them 
witli matter taken from the medulla of a horse and of a 
mule, both of which had died from traumatic tetanus. 
These uniform observations leave no room to doubt that 
tetanus is often, at least, due to a germ which exists in 
many places in the soil, and that the disease is transmissible 
by inoculation. 
Bonome observed nine cases of tetanus among seventy 
persons injured by the falling of a church from the 
earthquake at Bajardo. The bacillus of Nicolaier was 
detected in the wounds, and animals inoculated with the 
lime-dust of the fallen building died of tetanus. Of many 
persons injured by the falling of another church at the 
same time, none had tetanus, and animals inoculated with 
the lime from this church suffered no inconvenience. 
The same experimenter found the bacillus in the wound 
of a sheep which died from tetanus after castration. 
Beumer found the tetanus bacillus in the sloughing 
tissue of the umbilical cord of a child which was taken ill 
on the sixth day after birth, and died four days later from 
tetanus. From this he concludes that tetanus neonatorum 
and “ earth tetanus ” are identical, and advises that the cord 
should be dressed antiseptically. 
Kitasato has succeeded in isolating the bacillus of 
Nicolaier by growing the mixed cultures, from the pus of a 
wound on a man who died from tetanus, at a high tempera- 
ture (80°), and subsequently developing the germ under 
hydrogen. The bacillus grows only in the absence of air, 
and not in carbonic acid. It develops on agar, blood-serum, 
and gelatin, the last of which it gradually liquefies witli the 
formation of gas. The growth is more vigorous when the 
nutritive medium contains from 1.5 to 2 per cent of grape- 
sugar. 
In 1888 Belfanti and Pescarolo found in the pus of 116 
bacterial poisons. 
a wound, which was followed by tetanus, a bacillus which 
they believed to differ morphologically from that of Nico- 
laier and Rosenbach, and which in pure cultures induces 
tetanus in animals. The number of animals experimented 
upon was great and included mice, guinea-pigs, frogs, 
rabbits, pigeons, geese, sparrows, a chicken, and a dog. 
The pigeons, chicken, geese, and frogs proved immune. 
After subcutaneous injections a bloody oedema appeared at 
the place of inoculation and pus formed in small quantity. 
Paralysis first appeared and was followed by convulsions 
and opisthotonos. Later studies lead Belfanti and Pes- 
carolo to conclude that their bacillus is really that of 
Nicolaier, but differing somewhat from that of Kita- 
sato. Kitasato states positively that the germ which he 
has isolated is absolutely anaerobic, while the Italians find 
that theirs will not only grow aerobically, but when so 
grown will induce a classical tetanus. 
Lampiasi found in the blood from various organs of a 
man who died from so-called spontaneous tetanus, and in 
two cases of tetanus in mules, a spore-forming bacillus, 
which in pure cultures induced tetanus in animals. This 
bacillus is wholly different morphologically from that of 
Nicolaier. 
Widenmann reports a very interesting wise of a boy 
who fell from a wall and wounded his face on a piece of 
vine-stake in the earth. The boy died of tetanus, and the 
splinters extracted from the face and the earth about the 
stake were examined. The splinter was introduced under 
the skin of a mouse, which died thirty hours later of tetanus. 
In the pus formed about the splinter numerous microorgan- 
isms, among which a micrococcus and a short, thick bacillus 
abounded, were found, but in none of the many animals ex- 
perimented upon could the bacillus of Nicolaier be de- 
tected. In animals inoculated with the earth, however, the 
Nicolaier germ was found. Widenmann concludes that 
the so-called tetanus bacillus is found in most cases on ac- 
count of its very wide distribution in the soil and not as a 
result of its causal relation to the disease. 
Flugge has produced tetanus in animals without being TETANUS. 
117 
able to find the bacillus of Nicolaier, and Wyssokow- 
itsch has examined an earth which did not induce tetanus, 
but which caused suppuration, and in the pus the Nico- 
laier bacillus was found to be abundant. With the pus 
obtained from three eases of tetanus neonatorum due to 
omphalitis Kischensky induced tetanus in animals. The 
pus contained pyogenetic micrococci and a short bacillus, 
but the germ of Nicolaier could not be detected. 
Although Kitt claims that his tetanus bacillus is iden- 
tical with that of Kitasato (which is now regarded as a 
pure culture of the germ of Nicolaier), the former lique- 
fies solid blood-serum and the latter does not. Bacteriolo- 
gists generally agree that the Nicolaier bacillus is found 
only at the place of inoculation and that it is never present 
in the blood or internal organs, yet Shakespeare, as we 
have seen, induced tetanus in rabbits by inoculating them 
with matter taken from the medulla of a horse and that of 
a mule, both of which had died of tetanus. The bacillus 
which has been so well studied by Tizzoni and Cattani 
has certain constant biological differences from that of 
Kitasato. 
Pla has studied eight cases of traumatic tetanus both by 
cultures and by inoculation of animals. In none has he 
found the germ of Nicolaier. Moreover, since tetanus 
was induced in animals by bits of matter taken from the 
spinal cord, the Nicolaier germ could not have been the 
cause, if, as bacteriologists now teach, this germ is never 
found save at the place of inoculation. 
Brieger has obtained in the mixed cultures of the germ 
of Nicolaier and Bosenbacii four poisonous substances. 
The first, tetanine, which rapidly decomposes in acid solu- 
tions, but is stable in alkaline solutions, produces tetanus 
in mice when injected in quantities of only a few milli- 
grammes. The second, tetanotoxine, produces first tremor, 
then paralysis followed by severe convulsions. The third, 
to which no name has been given, causes tetanus accom- 
panied by free flow of the saliva and tears. The fourth, 
spasmotoxine, induces heavy clonic and tonic convulsions. 
Brieger has also isolated tetanine from the amputated 118 
BACTERIAL poisons. 
arm of a man with tetanus, thus showing that this chemical 
poison is formed in the body as well as in the artificial 
cultures. 
Brieger and Frankel obtained a “ toxalbumin ” from 
a culture of Kitasato’s germ in bouillon containing grape- 
sugar. This substance is soluble in water, and when in- 
jected in small amounts subcutaneously in guinea-pigs, 
tetanus appears in about four days, and soon terminates 
fatally. On the other hand, cultures of the bacillus of 
Tizzoni and Cattani in bouillon with sugar fail to pro- 
duce any chemical poison, but the cultures in gelatin are 
highly poisonous after filtration through porcelain. Even 
one-half cubic centimetre of the latter induces the disease 
and death in rabbits weighing from one and a half to two 
kilogrammes. Death results never later than three days, 
while, as has been seen above, the first symptoms induced 
by the poison from the bacillus of Kitasato usually 
appear on the fourth day. Brieger and Frankel ob- 
tained their proteid by precipitation with absolute alcohol, 
but the addition of this agent to cultures of the germ of 
Tizzoni and Cattani destroys its poisonous properties. 
The active substance of the Italian germ was obtained 
either (1) by dialysis, solution in water, and evaporation in 
a vacuum; or (2) by precipitation with ammonium sul- 
phate, separation by dialysis, and drying in a vacuum. 
This poisonous body is soluble in water, non-dialyzable, 
destructible by a temperature above 60°, and by treatment 
with concentrated mineral acids, and is unaffected by alka- 
lies or by prolonged treatment with carbonic acid gas. It 
contains a ferment which liquefies gelatin and digests fibrin. 
This peptonizing ferment is active only in alkaline solu- 
tion, and is present in the bouillon cultures which are not 
poisonous; therefore, the poison and the peptonizing fer- 
ment must be two distinct bodies. However, on account 
of the properties which we have mentioned, Tizzoni and 
Cattani conclude that, the poison also belongs to the 
soluble ferments or enzymes. 
Buschettini has studied the distribution of this poison tetanus. 
119 
through the body and its elimination in the following 
manner: 
Animals were poisoned by injections of the substance 
prepared by Tizzoni and Cattani, and just before death 
they were killed and bits of various organs rubbed up with 
sterilized water were injected into other animals. Emul- 
sions from the liver and supra-renal capsules were invariably 
without effect, while those from the kidney were constantly 
poisonous. This is supposed to prove that the poison is 
eliminated by the kidney. The blood taken from the vena 
cava was found to be poisonous in three out of four experi- 
ments. When the injections were made under the skin 
the lumbar cord was active in four out of eight cases, and 
in all, when the injections were made directly into the 
sciatic nerve. On the other hand, when the inoculations 
were made under the dura mater, the brain was found to 
be active while the lumbar cord remained inactive. From 
these experiments it is concluded that the poison not only 
circulates in the blood, but is deposited in the central 
nervous system. 
A. Babes prepared, from cultures made by Y. Babes 
and Puscaria in agar containing no peptone, an albumose 
which causes tetanus in animals. 
Faber finds in a mixed culture a poisonous proteid 
body which resembles closely, so far as it has been studied, 
that of Tizzoni and Cattani. Faber lays much stress 
upon the arguments in favor of this substance being a 
soluble ferment. With this proteid, convulsive movements 
first appear and become very distinct in the muscles about 
the point of injection. In case very small amounts are 
employed, the convulsive movements do not become general 
and the animal finally recovers. 
Peyraud claims to have secured immunity in animals 
against “ earth tetanus ” by giving to them strychnia in 
gradually increased doses. Nocard could not confirm this 
claim. 
According to Led antes, the poisonous arrows of the 
natives of the New Hebrides are prepared as follows : The 
points, which are usually made from human bones, are first 120 
BACTERIAL POISONS. 
covered with a vegetable resin, then smeared with the slime 
of swampy places. 
Ljermann found that material taken from the arm of a 
man who had died from tetanus, and who had been buried 
for two and one-half years, induced tetanus in animals. 
Tin’s would seem to show that the poison retains its viru- 
lence for a long time. In this material there were found 
nine kinds of bacteria, but none of these in pure culture, 
or in mixed culture, induced the disease. This is explained 
l>y the supposition that non-pathogenie bacteria may receive 
toxicogenie properties from the media in which they grow. 
Tuberculosis.—Whatever may be the ultimate verdict 
concerning the curative properties of Koch’s tuberculin, its 
employment has made us familiar with the action of the 
chemical products of the bacillus tuberculosis on man. Un- 
fortunately, Koch has given us but little information con- 
cerning the nature of his tuberculin, and the little which he 
has given us has been to some extent misleading. We 
would not imply that he has intentionally been misleading. 
Indeed, we believe that such avus not his intention. He 
speaks of the agent as an extract of a pure culture of the 
bacillus tuberculosis with 50 per cent, glycerin. One would 
infer from Koch’s statements that tuberculin is prepared 
by extracting the bacterial cells with 50 per cent, glycerin, 
and that the bacterial products are not present. But, 
as has been shown by IIueppe and Scholl, the proteids 
of the cells of the bacillus tuberculosis cannot be extracted 
with 50 per cent, glycerin. Moreover, the same investiga- 
tors have prepared a fluid identical in physical properties, 
in chemical reactions, and in its effects on animals, with 
Koch’s fluid, by each of the three following methods : 
1. Cultures of the bacillus are filtered, sterilizedjty heat, 
and concentrated. 
2. The supernatant, fluid portion of the culture is de- 
canted from the mass of germs at the bottom of the flask, 
and then concentrated. 
3. The culture is freed from germs by filtration through 
a Chamberland filter, and concentrated. TUBERCULOSIS. 
121 
These fluids contain : 1, the constituents of the nutritive 
medium which have not been altered by the growth of the 
germ, such as glycerin, albumins, albumoses, and peptones ; 
2, the bacterial products, which may possibly belong to the 
ptomaines, the bacterial albumins or albumoses and bacterial 
ferments; and 3, any constituents of dead, broken-down 
bacilli which may have passed into solution. To which of 
these constituents the action of the fluid is due has not been 
positively determined. However, from the similarity in 
the action of this fluid with that of the bacterial products 
of other germs, we seem justified in assuming that these 
constitute the active principle. 
As early as 1888, Hammersciilag found a poisonous 
proteid among the products of the growth of this germ. 
More recently he finds that as much as 27 per cent, of the 
cellular substance of the bacillus tuberculosis is soluble in 
alcohol and ether. In this extract there is, in addition to 
fat and lecithin, a poison which induces in rabbits and 
guinea-pigs convulsions followed by death. The part insol- 
uble in alcohol and ether consists of cellulose and proteids. 
Hammerschlag has also prepared from cultures of this 
bacillus a “ toxalbumin ” which, when injected subcuta- 
neously in rabbits, causes an elevation of temperature of 
from 1° to 2°, which continues for a day or longer. 
Zuelzer has reported the isolation of a poisonous pto- 
maine from agar cultures of the bacillus tuberculosis. He 
says that the injection of 1 centigramme or less of this 
substance subcutaneously in rabbits or guinea-pigs causes, 
after from three to five minutes, increased frequency of 
respiration (to 180 per minute?) and an elevation of tem- 
perature of from 0.5° to 1°. He also reports marked pro- 
trusio bulbi as a constant symptom ; the eyes become very 
bright and the pupils are dilated. From two to three 
centigrammes suffice to kill rabbits, death occurring in from 
two to four days. The place of injection is reddened, and 
hemorrhagic spots are formed in the mucous membrane of 
the stomach and small intestines. In two instances from 
15 to 20 cubic'centiinetres of clear fluid were found in the 
peritoneal cavity. 122 
BACTERIAL POISONS. 
Baumgarten draws the following conclusions from his 
experiments with tuberculin on rabbits with inoculation 
tuberculosis : 
It causes an exudative inflammation in the vascular 
tissue about the tubercle, and in this way the tuberculous 
tissue may be isolated and, when situated superficially, re- 
moved. In some cases, however, after the prolonged employ- 
ment of the agent, the tuberculous tissue itself may, under 
the influence of the exudative fluid and the polynuclear 
leucocytes, break down and form abscesses. The bacilli 
themselves are in no way harmed by the use of tuber- 
culin, and, after its constant employment for months, they 
retain their original form and lose none of their virulence. 
Some preparations seem to show that the bacilli multiply 
more rapidly when the injections are made, but a positive 
statement on this point is reserved until further studies 
have been made. It is certain, however, that the non- 
tubercular tissue of animals acquires no immunity against 
the disease from the injections. This is shown by the 
appearance of metastatic foci in animals in which from 
seven to twelve grammes of the original amount 
which would be equivalent to from seventy to one hundred 
and eighty grammes in man) has been injected. It is further 
shown bv the fact that in some animals treated subcutane- 
ously, tubercles have appeared at the point of injection. 
Prudden and Hodenpate summarize the results which 
they have obtained by the inoculation of animals with dead 
tubercle bacilli as follows: “ These dead tubercle bacilli 
are markedly chemotactic. When introduced in consider- 
able amount into the subcutaneous tissue or into the pleural 
or abdominal cavities, they are distinctly pyogenetic, caus- 
ing aseptic localized suppuration. Under these conditions 
they are capable, moreover, of stimulating the tissues about 
the suppurative foci to the development of a new tissue, 
closely resembling the diffuse tubercle tissue induced by 
the living germ. We have found that dead tubercle bacilli 
introduced in small numbers into the bloodvessels of the 
rabbit largely disappear within a few hours or days, but 
that scattering individuals and clusters may remain here TUBERCULOSIS. 
123 
and there in the lungs and liver, clinging to the vessel walls 
for many days without inducing any marked changes in the 
latter. After a time, however—earliest in the lung, later, 
as a rule, in the liver—a cell proliferation occurs in the 
vicinity of these dead germs, which leads to the formation 
of new multiple nodular structures bearing a striking mor- 
phological resemblance to miliary tubercles. There is in 
them, however, no tendency to cheesy degeneration and no 
evidence of proliferation of the bacilli, but rather a steady 
diminution in their number. It seems to us that the new 
structures originate in a proliferation of the vascular endo- 
thelium under the stimulus of the dead and disintegrating 
germs.” 
Maffucci finds that cultures of the tubercle bacillus 
(from a mammal), when grown from one to six months on 
glycerin, blood-serum, or liquid blood-serum, and then 
sterilized by being repeatedly heated to from 65° to 70°, 
produces in guinea-pigs, when employed subcutaneously, a 
progressive marasmus, which terminates fatally within from 
fourteen days to five or six months. lie also finds that 
eggs inoculated with sterilized cultures of the chicken tuber- 
culosis bacillus produce chickens which are feeble and soon 
die of emaciation. In neither the guinea-pigs nor chickens 
could he find any tubercles. This author, unfortunately, 
does not state positively whether the bacilli employed in his 
experiments on guinea-pigs were obtained from man or some 
other mammal. 
Crookshank and Herroun report the isolation of a 
ptomaine and an albumose not only from artificial cultures 
of the bacillus, but also from bovine tuberculous tissue. 
The ptomaine is reported as causing an elevation of tem- 
perature in tuberculous, and a depression in healthy, ani- 
mals. “ The albumose, whether obtained from pure culti- 
vations of the bacillus, or from tuberculous tissue, produced 
a marked rise of temperature in tuberculous guinea-pigs. 
On the other hand, in an experiment tried on a healthy 
guinea-pig, there was an equally well-marked fall of tem- 
perature.” 124 
BACTERIAL POISONS. 
Diphtheria.—That the Loffler bacillus is a cause 
of diphtheria no one can now deny. The fact that this 
germ, although found only at the seat of inoculation, causes 
marked systemic disturbances, indicates that its action must 
be due to its soluble products. This was early recognized 
by Loffler, who in 1887 attempted to ascertain the 
nature of the poison. A flask of bouillon containing pep- 
tone and grape-sugar was, three days after it had been 
inoculated with the bacillus, evaporated to 10 c.c., and this 
was injected into an animal, but was without effect. A 
second flask of the same material was extracted with ether, 
but this extract was also found to be inert. Next, some 
neutral beef broth was extracted with glycerin some four 
or five days after it had been inoculated with the bacillus. 
The glycerin extract, when treated with five times its 
volume of absolute alcohol, deposited a voluminous, floc- 
culent precipitate, which was collected, washed with alcohol, 
dried, and dissolved in a little water. A further precipita- 
tion with alcohol and a current of carbonic acid gas secured 
a white substance, and the injection of from 0.1 to 0.2 
gramme of this, dissolved in water, subcutaneously in 
guinea-pigs, caused marked pain followed by a fibrous 
swelling with hemorrhage into the muscles and oedema, 
terminating in necrosis. From these studies Loffler 
concluded that the poison belongs to the enzymes. 
Roux and Yersin found that bouillon cultures from 
which the bacillus had been removed by filtration through 
a Chamberland filter are poisonous, especially cultures 
which are four or five weeks old. The results obtained 
varied with the amount of the fluid, the species of animal, 
and the method of administration. The effects observed 
were a serous exudation into the pleural cavity, a marked, 
acute inflammation of the kidney, fatty degeneration of the 
liver, especially after injection into a bloodvessel, and (ede- 
matous swelling in the surrounding tissue after subcu- 
taneous inoculation. In some instances, in dogs, rabbits, 
and guinea-pigs, paralysis, generally in the posterior extre- 
mities, followed. The action of the poison was found to 
be very slow, and, as a rule, death occurred days, and in DIPHTHERIA. 
125 
some instances weeks, after the inoculation, and was pre- 
ceded by marked emaciation. 
The cultures first employed were seven days old; older 
cultures (six weeks) contain more of the poison, and the 
symptoms appear within a few hours. In cultures espe- 
cially rich in the poison, a small amount (from 0.2 to 2 c.e.) 
injected under the skin in guinea-pigs suffices to induce the 
symptoms. Mice and rats are markedly insusceptible, but 
succumb to large doses. 
Heating to 100° for twenty minutes renders the poison 
inert, and a temperature of 58° maintained for two hours 
markedly lessens its virulence. 
The poisonous substance is precipitated by absolute 
alcohol, and is carried down mechanically on the addi- 
tion of calcium chloride to the filtered cultures. These 
investigators agree with Loffler that the poison belongs 
to the enzymes. The great toxicity of this substance is 
indicated by the statement of Roux and Yersin that 0.4 
milligramme suffices to kill eight guinea-pigs or two rab- 
bits, and that 2 centigrammes of the calcium chloride 
precipitate, containing about 0.2 milligramme of the pure 
poison, will kill a guinea-pig within four days. 
Brieger and Franker have made a very complete 
study of the chemical products of the Loffler bacillus. 
They employed cultures of bouillon and peptone containing 
from five to six per cent, of glycerin, and others containing 
ten per cent, of sterile, fluid blood-serum. The latter were 
found to be most suitable. In these the bacilli grow most 
abundantly. In all cases they confirmed the statement of 
Roux and Yersin that the cultures, at first alkaline, be- 
come strongly acid, and finally again alkaline, with the 
exception that the glycerin cultures remained acid. 
For the removal of the bacteria two methods were em- 
ployed. First the bacilli were destroyed by heat. When 
a temperature of 100° was employed the cultures were 
rendered inert, but it was found that exposure for from 
three to four hours to a temperature of 50° was sufficient 
to destroy the germs, while the virulence of the chemical 
products was not affected. The second method of removing 126 
bacterial poisons. 
the bacteria consisted of filtration through a Chamberland 
filter. The germ-free filtrate could be heated to 50° with- 
out loss of toxicity, while a temperature of 60° rendered 
it inert. In the majority of the experiments the filtration 
method was used and in this way a large quantity of a 
poisonous fluid of uniform strength was obtained. 
Varying amounts of this fluid were used upon animals, 
mostly guinea-pigs and rabbits, and it was found that the 
effects varied with the quantities employed and the methods 
of administration. The symptoms appeared most promptly 
when the injections were made directly into a bloodvessel. 
Of four rabbits which were given subcutaneously respec- 
tively 1, 21, 5, and 10 c.c. of the filtrate on December the 
28th, the first died January 4th ; the second, January 2d ; 
the third, December 31st; and the fourth, December 30th. 
In all cases in which death did not occur too early, paralysis 
appeared. The limbs were first paralyzed, and this was 
true whether the fluid was administered intravenously or 
subcutaneously. The post-mortem appearances were iden- 
tical with those observed after inoculation with the bacillus, 
with the exception of the absence of the pseudo-membrane. 
After subcutaneous injection there was a gelatinous, grayish- 
white, sometimes reddish, cedematous fluid formed at the 
point of injection; and, after larger doses, necrosis. In 
cases in which death was delayed, there were effusions in 
the pleura, fatty degeneration of the liver, and inflamma- 
tion of the kidneys. Especially marked were these cellular 
changes in rabbits which were treated with small amounts 
intravenously. 
Brieger and Franker conclude this part of their 
report with the following statement: “ We have shown 
that the Loffler diphtheria bacillus produces in its cul- 
tures a poisonous, soluble substance, separable from the 
bacteria, which causes in susceptible animals the same 
phenomena which are induced by inoculation with the 
living microorganism. We have further shown that this 
substance is destroyed by a temperature over 60°, but that 
it can be heated to 50°, even in the presence of an excess 
of hydrochloric acid, without being destroyed. This last DIPHTHERIA. 
127 
fact is contrary to the assumption that the chemical poison 
of the diphtheria bacillus is a ferment or enzyme.” 
The fluid was tested for basic products, but with wholly 
negative results, except that small amounts of kreatinin and 
cholin were found. It was also distilled at from 20° to 
35° in a vacuum, and the distillate was found to be inert. 
The poisonous substance was found to be insoluble in 
alcohol, soluble in water, and non-dialyzable. It was pre- 
cipitated by saturation witli ammonium sulphate. 
The substance was obtained by allowing the germ-free 
filtrate, after being rendered feebly acid with acetic acid, to 
fall into a large volume of absolute alcohol. It was puri- 
fied by repeated solution in water and precipitation with 
alcohol. It contains a large amount of sulphur, and re- 
sponds to the biuret and Millon tests. It is, therefore, 
classified among the albumins. Since it is not precipitated 
by saturation with magnesium sulphate at 30°, it cannot 
belong to the globulins. The fact that it is precipitated by 
saturation with ammonium sulphate, and that it does not 
dialyze, shows that it is not peptone. It is, therefore, 
classified by Brieger and Frankel among the albumins, 
and is designated as a “ toxalbumin.” 
The special reactions and the results of an ultimate 
analysis of this substance have already been given (page 20). 
This proteid induces in animals all the symptoms and 
post-mortem appearances which have been mentioned as 
following the administration of the filtered cultures. It is 
to be noted that the injection of small quantities of this 
proteid (2|- milligrammes per 1 kilogramme of the body- 
weight of the animal) does not produce its effects until after 
the lapse of weeks, and possibly months. This peculiarity 
in action distinguishes this class of substances from all other 
chemical poisons, and it has received as yet no satisfactory 
explanation. There is no reason for believing that the body 
obtained by Brieger and Frankel is chemically pure, 
and until it has been obtained in this condition we can 
only speculate concerning its true nature. 
It should be remarked that the Loffler bacillus shows 
not only marked morphological variations, but that it is 128 
BACTERIAL POISONS. 
very variable in its virulence, some cultures having been 
obtained which are wholly without effect upon animals. 
From cultures of this kind Brieger and Franker pre- 
pared a non-poisonous albumin differing in its ultimate 
composition and in many of its chemical reactions from the 
poisonous one. 
Frankel has been unable to secure immunity in ani- 
mals against diphtheria by the employment of small doses 
of the “ toxalbumin.” If the dose is large enough the 
animal dies. If it is smaller, the animal seems to become 
more susceptible and succumbs more readily to inoculations 
with the germ. While this is true of the filtered culture, it 
is not the case with that which has been sterilized by heat. 
Frankel finds that if from 10 to 20 c.c. of a cul- 
ture of the bacillus three weeks old, which has been 
heated for one hour at from 65° to 70°, be injected under 
the skin of the abdomen of guinea-pigs, immunity against 
subsequent inoculation with the virulent germ is secured, 
provided that the inoculation is not made earlier than the 
fourteenth day after the treatment with the sterilized 
culture. He thinks that the culture contains two specific 
albumins, one of which is poisonous, while the other gives 
immunity. The former is destroyed by a temperature of 
from 65° to 70°, while the other retains its characteristic 
properties. He admits the possibility that the poisonous 
albumin may be converted into the other form by the high 
temperature. He finds that the modified culture, which 
gives immunity, is of no service for therapeutic purposes, 
and that if an animal be treated with it directly after inocu- 
lation with the germ, death is not retarded, but is hastened. 
From these experiments he concludes that the vaccination 
albumin at first lessens, and subsequently increases the 
resistance of the animal. 
Sprouck and his students have confirmed the above 
statements concerning the toxicity of the germ-free cultures 
of this bacillus. They have also called attention to the 
albuminuria following the employment of this poison. In 
the urine they find casts, white, and sometimes red, blood- 
corpuscles. Microscopic examination of the kidney after SUPPURATION. 
129 
death shows the same changes which are observed in the 
diphtheritic nephritis of children. Babes also finds that 
the germ-free cultures produce the parenchymatous degener- 
ations of the internal organs which arc found in the human 
body. 
Tange has shown that the chemical poison is formed in 
the body as well as in culture-flasks. A large piece of 
pseudo-membrane was macerated in water in an ice-chest 
for twenty-four hours, and then filtered through porcelain. 
The filtrate, injected into animals, produced all the symp- 
toms which have been obtained by a similar employment of 
artificial cultures. Tangl also observed that in some cases 
in which the animals were inoculated with the sterilized 
culture through the mucous membrane a pseudo-membrane 
formed at the point of injection. 
Suppuration.—As early as 1879, Leber concluded 
from his observation on infective keratitis that the asper- 
g’illus must produce certain soluble products which diffuse 
through the cornea and set up an inflammatory action in 
the adjacent vascular tissue. In 1882, he showed that sup- 
puration could be induced by the introduction of sterilized 
mercury and copper, and that the pus formed is free from 
germs. In 1884, lie induced suppuration by the injection 
of cultures of the staphylococcus pyogenes aureus which 
had been sterilized by being boiled for hours. In 1888, 
the same investigator reported that he had found an alco- 
holic extract of the dried staphylococcus to be highly pyo- 
genetic. From this extract he has prepared a crystalline 
body which he calls phlogosin. This substance is readily 
soluble in alcohol and ether,.sparingly soluble in water, 
and it crystallizes in needles. The crystals can be sub- 
limed, leaving no residue, and the sublimate, which forms 
in rosettes, still possesses the pyogeneticproperties. Alkalies 
precipitate this substance from its solution in amorphous 
granules, which dissolve in acids, forming crystalline salts. 
Leber refers to the observation of the botanist Pfeffer, 
who found that vegetable cells are attracted by certain 
chemical substances, and adopts the term chemotactic action 130 
BACTERIAL POISONS. 
(chemotactische Wirkung) to indicate the property of certain 
chemical agents of attracting leucocytes. 
As has been stated, Buchner has found that the cells of 
many bacteria contain pyogenetic proteids. The amount 
of these substances in the cells varies with the kind of 
germ, and some species (the bacillus prodigiosus, for in- 
stance) seem to contain no such bodies. The bacillus pyo- 
cyaneus contains a large quantity of the proteid, and is 
suitable for lecture demonstration. The germs are taken 
from potato cultures and rubbed up with water. Then 
they are treated with about fifty volumes of a 0.5 per cent, 
solution of caustic potash. This forms in the cold a muci- 
laginous mass which dissolves at the temperature of the 
water-bath. After being heated for some hours the fluid 
is filtered through a number of small filters; the first por- 
tions should be refiltered. The filtrate is a greenish fluid 
(pyocyanin) which by the careful addition of acetic or 
hydrochloric acid (an excess is to be avoided) forms a 
voluminous precipitate (pyocyaneus proteid). This pre- 
cipitate should be collected on a filter, washed with water, 
then suspended in water and a few drops of a soda solution 
added, when a dark-brown fluid, with a tendency to gela- 
tinize in the cold, containing about 10 per cent, of the pro- 
teid, is obtained. 
15.254 grammes of the moist bacteria yield 1.44 gramme 
of dry bacterial substance, and this after the treatment 
given above furnishes 0.2739 gramme of dry proteid = 
19.3 per cent. This proteid leaves 11.52 per cent, of ash, 
which contains phosphoric acid, but consists principally of 
sodium chloride. 
Much smaller amounts of proteid were obtained from 
other germs, but the Eberth germ, bacillus subtilis, lactic 
acid bacillus, red bacillus from potato, and staphylococcus 
Pyogenes aureus furnished considerable quantities. 
The chemotactic properties of these proteids were tested 
in the following manner : The dissolved proteid was placed 
in a spindle-shaped glass tube, and the tubes, sterilized by 
prolonged boiling, were introduced under the skin on the SUPPURATION. 
131 
backs of rabbits with antiseptic precautions, and the ends 
of the tubes broken off subcutaneously. 
After from two to three days the tubes were removed 
and found to contain, in addition to some of the proteid, 
several millimetres of fibrinous pus, which was examined 
microscopically and by the preparations of cultures, which 
invariably remained sterile. The proteid of the Eberth 
bacillus was found to have specially marked pyogenetic 
properties. 
Similar experiments were made with the following crys- 
talline substances: the butyrate and valerianate of ammo- 
nia (each 1 per cent, solution), trimethylamin (2 per cent.), 
ammonia (2 per cent.), leucin, tyrosin and glycocol (1 per 
cent.), urea (5 per cent.), and urate of ammonia and skatol 
(1 per cent.). Glycocol and leucin only were found to have 
the chemotactic action, and with these this action was but 
slight compared with that of the bacterial proteids. 
The next experiments were made with the object of 
ascertaining whether or not proteids similar to those derived 
from the bacteria would cause a like effect. The bacterial 
cellular proteids resemble very closely vegetable casein 
some of which was prepared from wheat gluten and tested 
as above. This proteid was found to be possessed of 
marked chemotactic properties. The subcutaneous injec- 
tion of sterilized preparations of wheat-flour and ground 
peas were also found to cause suppuration. Negative 
results were obtained witli starch and solutions of disodium 
hydrie phosphate. From this it is concluded that the active 
agent in the flour is its casein. 
Peptone was employed without effect, while gelatin was 
found to act energetically. Alkaline albuminates were 
prepared from muscle, liver, lungs, and kidney by treating 
finely divided portions of these organs with potash and pro- 
ceeding as in the preparation of the bacterial proteids. All 
of these caused the formation of pus, and the preparations 
from the liver were found to be specially potent. 
Similar preparations from blood and egg-yolk were 
active, while those from fibrin and the white of egg had no 
effect. Hemi-albumose was also found to be active, and 132 
BACTERIAL POISONS. 
this fact is placed in contrast with the negative result 
obtained with peptone. 
One of the most interesting results was obtained by the 
daily injection of a chemotactic proteid directly into the 
blood. Before the first injection the proportion of white to 
red corpuscles was 1 : 318 ; on the second day, 1 : 126 ; on 
the third, 1 :102 ; on the morning of the fourth, 1 : 73 ; on 
the afternoon of the fourth, 1 :38. After this there was 
no further increase. The absolute number of red corpuscles 
remained unchanged, while the absolute number of the 
white multiplied sevenfold. The white corpuscles were on 
the first days often found in groups of from two to four, 
and later, of from ten to twenty. This seems to demon- 
strate that these substances cause an increased production 
of leucocytes. General leucocytosis was induced by the 
similar employment of vegetable casein and an alkaline 
albuminate prepared from the muscles of a calf. 
Finally, Buchner tested the action of this proteid upon 
himself. One cubic centimetre of a very dilute solution, 
containing 3.5 milligrammes of the solid proteid, was 
injected under the skin of the forearm with antiseptic pre- 
cautions. Two hours later there was marked pain along 
the lymphatics, especially localized in the elbow and axilla. 
The temperature showed no marked elevation (only 37.8°). 
On the following day there were marked erysipelatous 
redness and swelling extending for some inches about the 
place of injection, and accompanied by severe pain. The 
inflamed area felt hot, and projected distinctly above the 
surrounding surface. The lymphatics of the arm appeared 
like red cords. On the third day the swelling and redness 
were more marked, and extended from the wrist to the 
elbow. On the fourth day the symptoms began to recede. 
Here we have clinically a perfectly typical erysipelas with 
lymphangitis, and Buchner claims that all the cardinal 
symptoms of inflammation—rubor, calor, dolor—could not 
be produced without involvement of the solid tissues. 
Similar, but loss marked, symptoms were induced by the 
injection of a dilute solution of vegetable casein. 
Buchner states that bacteria will not cause infamma- SUMMER DIARRH(EAS OF INFANCY. 
133 
tion unless they be broken down. The pyogenetic substance 
contained within the bacterial cell can have no chemotactic 
action until the cell disintegrates. Thus, the anthrax 
bacillus contains a pyogenetic substance, but no pus is 
formed in mice with anthrax, because there is no destruction 
of the bacilli. This pyogenetic proteid of the anthrax 
bacillus, however, manifests its action in malignant pustule. 
These experiments are of the greatest interest. We must 
say, however, that it is possible that the bacterial cellular 
proteid may be modified by the treatment to which it has 
been subjected in these experiments. We do not as yet 
know enough about the nature of this proteid to say that 
its nature and its action are not altered by being heated for 
hours with an alkali. However, accepting Buchner’s 
work, it throws much light upon processes which have 
heretofore been but imperfectly understood. 
The Summer Diarrhoeas of Infancy.—In a paper 
published in 1888, Vaughan stated that the microorgan- 
isms which produce the catarrhal or mucous diarrhoeas of 
infancy are probably only putrefactive or saprophytic in 
(“haracter, and that they prove harmful by splitting up 
complex molecules and forming chemical poisons. At that 
time it was generally believed that a specific germ would 
be found, but the truth of the above statement is being 
made more manifest with every experimental study of the 
subject. Able and diligent bacteriologists, among whom 
Booker, in this country, and Escherich, in Germany, 
deserve special mention, have made a careful study of the 
bacteria found in the intestines and stools in these diseases, 
and all agree that no specific organism has been found. 
Booker has reported the isolation of more than thirty 
kinds. In true cholera infantum the proteus group of bac- 
teria was found in fifteen out of nineteen cases, but in the 
ordinary diarrhoeas there is no constancy in the species 
present. Germs which are frequently found one year are 
rarely seen in the cases observed the next summer. This 
has been the experience of all who have studied the bacteria 
of the summer diarrhoeas of infancy. Vaughan has studied 134 
BACTERIAL POISONS. 
the chemical products of the germs x, a, and A of Booker’s 
list in the following manner and with the results as stated 
below. 
Of these germs, Booker makes the following statements : 
“x was found almost as a pure culture in the feces of a 
fatal ease of diarrhoea, a was strongly pathogenic when 
tested last winter. A was isolated last summer; liquefies 
gelatin, and belongs to the proteus group.” 
Beef-tea cultures of each of these germs were made and 
kept in an incubator at 37° for forty-eight hours. At the ex- 
piration of this time these cultures were used for inoculating 
flasks of sterilized beef-broth. Eight flasks, each contain- 
ing about ten ounces, were employed for each germ. These 
•cultures were kept in the incubator at 37° for ten days. 
They were then twice filtered through heavy Swedish filter- 
paper. The second filtrate was allowed to fall into a large 
volume of absolute alcohol feebly acidified with acetic acid. 
A voluminous, flocculent precipitate resulted in each case. 
After the precipitates had subsided the supernatant fluid 
was decanted. The precipitates were then treated with dis- 
tilled water, in which those from x and a were soluble, 
while that from A proved insoluble. A large volume of 
absolute alcohol was again added, and the mixture allowed 
to stand for four days. The precipitates from x and a com- 
pletely subsided, leaving the supernatant fluids perfectly 
clear; but in the case of A the subsidence was not com- 
plete. The precipitates were collected, by decantation and 
filtration, on porous plates, and dried over sulphuric acid. 
These substances are proteid in composition, but differ f rom 
known proteids and from one another That from x is 
slightly yellow, as seen deposited in the alcohol, but be- 
comes grayish on exposure to the air. It is readily soluble 
in water, from which it is not precipitated by heat or nitric 
acid, singly or combined. 
It gives the biuret and xantho-proteid reactions. It is 
precipitated by saturating its aqueous solution with ammo- 
nium sulphate, and therefore cannot be classed with the 
peptones. Sodium sulphate and carbonic acid fail to throw SUMMER DIARRHOEAS OF INFANCY. 
135 
it down from its aqueous solution, consequently we must 
say that it is not a globulin. 
This leaves us with no other choice than to place it 
among the albumins, but we must admit that it possesses 
properties which do not belong to the known albumins. 
The proteid prepared from cultures of the germ a is, as 
seen under the alcohol, very light, flocculent, and perfectly 
white, but so soon as it is brought in contact with the air 
it begins to blacken, and finally dries down on the porous 
plate in black scales. 
It possesses the same general properties in regard to the 
action of solvents and other reagents which were found to 
be possessed by the proteid obtained from cultures of x. 
The proteid of A is peculiar, inasmuch as it is practically 
insoluble in water. 
These three proteids are highly poisonous. When in- 
jected under the skin of kittens or dogs they cause vomit- 
ing and purging, and, when employed in sufficient quantity, 
collapse and death. Post-mortem examination shows the 
small intestine pale throughout and constricted in places. 
The heart has been invariably, so far, found in diastole and 
filled with blood. The following brief notes from the 
record of experiments will illustrate the nature of the 
symptoms and the post-mortem appearances. 
A small amount of proteid from bacillus x, dissolved in 
water, was injected under the skin on the back of a kitten 
about eight weeks old. Within one-half hour the animal 
began to vomit and purge, and death resulted within eigh- 
teen hours. The small intestines were pale, contracted in 
places, and contained a frothy mucus. The stomach was 
distended with gas and contained yellowish mucus. The 
liver was normal, the spleen and kidneys congested, and 
the heart distended. 
Another kitten was treated with the proteid from bacil- 
lus a, dissolved in water. The vomited and fecal matters 
in this case were green. The animal died after fifteen 
hours, and presented appearances practically identical with 
those mentioned above. 
A third kitten was treated with some of the proteid of 136 
BACTERIAL POISONS. 
bacillus A, suspended in water, and presented substantially 
the same symptoms and post-mortem appearances. 
A fourth animal was treated in the same manner as the 
above with a proteid prepared from some canned meat. 
This was done as a control on tlie above experiments, and 
the kitten remained unaffected. This experiment demon- 
strates the fact that the poisonous properties are peculiar to 
the bacterial proteids. 
Concerning the amount of one of these proteids neces- 
sary to produce a fatal result in the animals experimented 
upon a few experiments have been made. 
Under the skin on the back of a guinea-pig, Vaughan 
injected ten milligrammes of the dry-scale proteid from 
bacillus a. This caused death within twelve hours. Of 
two kittens treated with fifteen milligrammes each of the 
a albumin, one died after forty-eight hours and the other 
recovered after two days of purging and vomiting. Two 
dogs, of about five pounds’ w’eight, had each forty milli- 
grammes, and, after serious illness of two days’ duration, 
speedily recovered. 
During these two days of vomiting and purging the dogs 
were constantly shivering, as with cold, but the rectal tem- 
perature stood at from 102.5° to 103.5° F. 
There was in no case any sign of inflammation at the 
point of injection. 
Plate cultures have been made from the proteids them- 
selves and from the blood, liver, spleen, and kidneys of 
some of the animals killed with the proteid, and these plates 
have remained sterile, thus demonstrating that no germ 
has been introduced into the animal along with the chem- 
ical poison. 
What conclusions may we draw from these facts when 
considered in connection with the results of the labors of 
Booker and Escherich? We will formulate our ideas 
in the following propositions: 
(1) There are many germs, any one of which, when in- 
troduced into the intestines of the infant, under certain 
favorable conditions, may produce diarrhoea. 
As has been stated, many different germs have been SUMMER DIARRHOEAS OF INFANCY. 
137 
found in the intestines of infants suffering from summer 
diarrhoea, and vve now find that three species of these are 
capable of producing chemical poisons, which induce effects 
substantially identical with the symptoms observed in the 
infants, and it is not unreasonable to suppose that many 
other of these germs produce similar poisons. 
(2) Many of these germs are probably truly saprophytic. 
A germ growing in the intestine does not necessarily 
feed upon living tissue. The food in the duodenum before 
absorption has no more vitality than the same material in 
the flask. Moreover, the excretions poured into the intes- 
tines from the body are not supposed to be possessed of 
vitality. A germ which will grow upon a certain medium 
in the flask and produce a poison will grow on the same 
medium in the intestine and produce the same poison, pro- 
vided it is not destroyed by some secretion of the body. 
(8) The only digestive secretion which is known to have 
any decided germicidal effect is the gastric juice; therefore, 
if the secretion be impaired there is at least the possibility 
that the living germ will pass on to the intestine, will there 
multiply, and will, if it be capable of so doing, elaborate a 
chemical poison which may be absorbed. 
There is no longer any doubt that the acid of the gastric 
juice has a marked germicidal effect upon many of the 
microorganisms. 
Vaughan has found that an exposure to a two-tenths 
per cent, solution of hydrochloric acid for half an hour will 
destroy Eberth’s germ and two poison-producing bacilli 
which he has isolated from drinking-water which was 
believed to have caused typhoid fever. Although the ger- 
micidal effect of this acid has not been tried on the bacteria 
under consideration, doubtless it will be found to be con- 
siderable. 
The chief reason why the breast-fed child has a better 
chance for life than the one fed upon cow’s milk lies in the 
fact that the former gets its food germ-free; but a second 
reasou is to be found in the larger amount of acid required 
to neutralize the cow’s milk, as has been pointed out by 138 
BACTERIAL POISONS. 
Escherich. The gastric juice is the physiological guard 
against infection by way of the intestines.1 
It is also possible that some of the secretions poured into 
the intestines have germicidal properties, or that the cells, 
in absorbing the poisonous proteids, may to a limited ex- 
tent so alter them that they are no longer poisonous, or 
that in a perfectly normal condition the liver may be able 
to prevent these poisons from entering the general circula- 
tion without change. These are all possibilities, which 
science at some time in the future will investigate. 
(4) Any germ which is capable of growing and produc- 
ing an absorbable poison in the intestine is a pathogenic 
germ. 
It is not necessary that a germ be capable of growing 
and causing disease and death when injected under the skin 
or into the blood in order to establish its right to rank with 
the pathogenic germs. In the blood the organism is acted 
upon by a wholly different fluid from that with which it 
is surrounded in the intestine, and the germicidal properties 
of the blood have been unquestionably demonstrated. 
(5) The proper classification of germs in regard to their 
relation to disease cannot be made from their morphology 
alone, but must depend largely upon the products of their 
growth. 
As has been stated, three microorganisms, differing suf- 
ficiently to be recognized as of different species, produce 
poisons, all of which induce vomiting and purging, and, 
when used in sufficient quantity, death. Morphologically 
these bacilli may not be closely related, but physiologically 
they are near akin. 
If these deductions be true, we will try to avoid the 
introduction into the alimentary canal, not only of the so- 
called specific pathogenic germs, but of all toxicogeuic 
microorganisms. 
1 It has been said that this statement cannot he true, because there are 
other acids whicli are more powerful germicides than hydrochloric acid, 
but there is no force in this argument. The question is not whether the 
stomach is supplied with the very best germicide, but whether it is sup- 
plied with any at a'l. The human eye may not be a perfect mechanism, 
but it is man’s only organ of vision. TYPHOID fever. 
139 
Baginsky and Stadthagen have obtained from cul- 
tures of the “ white liquefying bacterium ” of the former a 
poisonous proteid which produces in mice, after about five 
hours, slight dyspnoea. The coat becomes rough, the ani- 
mal sits with drooping head, and when forced to move does 
so sluggishly, but without any evidence of paralysis. The 
marked apathy increases, and death results after two or 
three days. Section shows an infiltration about the place 
of injection, congestion of the spleen, liver, and peritoneum. 
The intestine is hypersemic throughout its entire length, 
and its upper portion contains a reddish-brown fluid. 
From cultures of the same bacterium Baginsky and 
Stadthagen have also obtained a poisonous ptomaine, 
which is probably identical with one found by Brieger 
in putrid horseflesh, and which has the formula C7H17N()2. 
That tyrotoxieon is one of the causes of the violent 
choleraic diarrhoea of children there can scarcely be a 
doubt. The symptoms induced by the poison cannot be 
distinguished from those of the disease. The post-mortem 
appearances are very much alike, if not identical, and the 
poison has been found in a sample of milk a part of which 
had been given to a child not more than two hours before 
the first symptoms of a violent attack of the disease made 
themselves manifest. 
Typhoid Fever. — Iu 1880, Eberth discovered a 
bacillus which he believed to be the cause of typhoid fever, 
and this belief has been quite generally accepted. In the 
first edition of this work it was stated that the fever and 
the characteristic lesions of the disease had been produced 
in animals by inoculation with this germ. This is now 
known to be erroneous. As has been stated (page 93), the 
essential lesions of typhoid fever may be produced in ani- 
mals with a number of microorganisms, among which, 
however, the Eberth bacillus is not included. The results 
obtained by Franker and Simmonds, and Seitz have been 
shown by Beumer and Peiper to be fallacious, and the 
germ with which the experiments were made by Vaughan 
and Novy, and mentioned in the first edition, is known 140 
BACTERIAL poisons. 
not to be identical with that of Eberth. It is true that 
this germ induced in dogs a continued fever of from 
twenty-eight to thirty-five days in duration, terminating 
in some instances fatally and revealing ulceration and per- 
foration of the small intestines, but for this reason it is 
known to be different from Eberth’s bacillus, because the 
latter never induces these effects. Notwithstanding this 
failure to affect the lower animals, the majority of bacteri- 
ologists believe, as lias been stated, that the Eberth 
bacillus is the sole and only cause of typhoid fever. In 
this believe Vaughan refuses to concur, and claims that 
the Eberth bacillus as found in the spleen after death is 
an involution form of any one of a number of germs 
which are found in certain waters. As this is not the 
place for an extended discussion of purely morphological 
questions, the reader is referred to the literature of the 
subject, and we will content ourselves with giving the 
following summary of what is known concerning the 
chemical products of the Eberth bacillus and of the 
germs studied by Vaughan. 
In 1885, Brieger obtained from pure cultures of the 
Eberth bacillus a poisonous ptomaine, which produced in 
guinea-pigs a slight flow of saliva, frequency of respira- 
tion, dilatation of the pupils, profuse diarrhoea, paralysis, 
and death within from twenty-four to forty-eight hours. 
Post-mortem examination showed the heart in systole, the 
lungs hypenemic, and the intestines contracted and pale. 
At first Brieger was inclined to regard this as the specific 
poison of typhoid fever and named it typhotoxine. How- 
ever, he has more recently modified his opinion and is 
inclined to regard typhoid fever as due to a mixed in- 
fection. 
Brieger and Franker have found in cultures of the 
Eberth bacillus a proteid which causes death in rabbits 
after from eight to ten days. They say nothing about the 
symptoms. 
In 1889, Vaughan isolated from mixed cultures from 
typhoid stools a base, forming crystalline salts and capable 
of inducing in cats and dogs a marked elevation of tern- TYPHOID FEVER. 
141 
perature accompanied by severe purging. The following 
is the record of one experiment with this substance: 
(i An aqueous solution of the crystals was given to a dog 
by the mouth at 3 p.m. The rectal temperature before the 
administration was 101° F. At 3.15, retching and vomit- 
ing set in and continued at intervals for more than two 
hours. At 3.30, the temperature was 103° F. At 3.55, 
the animal began to purge. The first discharges contained 
much fecal matter, but subsequently they were watery 
and contained mucus plainly stained with blood. At 4, 
the temperature was l03.5° F. and remained the same at 
4.30. The animal was not seen again until 10 a.m. the 
next day, when its temperature was 100.5°, and recovery 
seemed complete.” 
This base was not obtained in quantity sufficient for an 
ultimate analysis. The platino-chloride crystallizes in fine 
rhombic prisms and the hydrochloride in long, delicate, 
red needles. The red color seems to be inherent to the 
substance and not due to impurities. The mercury and 
platinum compounds are insoluble in alcohol, soluble in 
water. The hydrochloride is soluble in both water and 
alcohol. 
In 1890, Vaughan reported the isolation, from water 
supposed to cause typhoid fever, of a number of toxi- 
cogenic germs. The chemical products of two of these 
have been studied. They belong to the proteids, and an 
analysis of one of them by Freer shows it to belong to the 
nucleins. These poisons are soluble in water, the opales- 
cent solution showing a distinctly acid reaction. They are 
not precipitated by heat or nitric acid singly or combined. 
They dissolve in nitric acid, forming a colorless solution, 
which becomes yellow on the addition of ammonia. They 
dissolve in caustic alkalies and the solution becomes purple 
on the addition of a dilute solution of copper sulphate. 
On white rats these poisons produce symptoms which are 
identical with those which follow inoculations with the 
living germs. The rat seems to shiver with cold and gives 
evidence of abdominal pain. It lies with its limbs flexed 
and head drawn down for a few seconds, then stretches out 142 
bacterial poisons. 
the limbs. It lies on the side for a short time, then sits 
with the head drawn under the body. 
Dogs shiver as with cold, but at the same time the rectal 
temperature is from one to four degrees above the normal. 
In some instances vomiting and purging have been induced. 
The following experiments seem to show that the poison 
accumulates in the nerve-centres : 
Two guinea-pigs were treated with hypodermic injec- 
tions of one of these poisons, the amount used being about 
ten times the dose which ordinarily proves fatal to these 
animals. Within twelve hours both were dead. Plate 
cultures made from the liver, spleen, blood, brain, and 
spinal cord remained sterile. Small quantities of the brain 
and cord were rubbed up in a sterilized dish with sterilized 
water, and two c. c. of the emulsion were injected under 
the skin of each of four guinea-pigs. These animals seemed 
to be very excitable the next day, throwing themselves 
about violently in the cages when slight noises were made 
near them. Within a period of from sixteen to twenty- 
four days all died. This experiment needs repetition, and 
it will be necessary to prepare and inject similar emulsions 
made from other organs before any positive conclusions can 
be drawn. 
In a study of fatal cases of typhoid fever at Bucharest 
Babes finds that the typical germ differs markedly from 
that of Eberth. 
Swine-plague, or Hog-cholera.—The researches of 
Loffler, Schutz, Lydtin, and Sohottelius in Europe, 
and of Billings and Salmon in this country, have demon- 
strated the existence among swine of at least three infectious 
diseases. These are— 
(1) Hog-erysipelas, or rouget of France, or Sclnveine- 
rothlauf of Germany. 
(2) German swine-plague, or Schweineseuche. 
(3) American swine-plague (Billings), or hog-cholera 
(Salmon). 
The first two of these are exclusively European diseases, 
and their chemical poisons have not been studied. SWINE-PLAGUE. 
143 
The American swine-plague is preeminently a disease of 
the digestive tract involving most markedly the large in- 
testine. It is the great swine disease of this country, and 
is probably present in England, where it is associated with 
other diseases under the name of swine-fever. A disease 
which was observed in Denmark and Sweden for the first 
time in 1888-89 and known as swine-pest or swine-diph- 
theria, has been shown by Selander, Frosch, and others 
to be identical with our swine-plague. In the summer of 
1889 France was visited by a swine disease, which is con- 
sidered by Cornil and Chantemesse to be identical with 
the German swine-plague, but which Rietsch and Jobert, 
after a comparative study of the microorganisms, pronounce 
as the American disease. In this country we have at pres- 
ent no positive demonstration of the existence of auy other 
infectious swine disease. The swine-plague of Salmon 
has been the subject of considerable discussion, but its ex- 
istence can hardly be said to be established. 
The following statements concerning the chemical poisons 
refer to the swine-plague of Billings or the hog-cholera 
of Salmon, which are only two names for one disease. 
In pure cultures of this bacillus Novy has found a poi- 
sonous base, which probably has the composition C10H26N2, 
and to which he has provisionally given the name, suso- 
toxine. One hundred milligrammes of the hydrochloride 
of this base causes in white rats convulsive tremors and 
death within one and one-half hours. Post-mortem exam- 
ination shows the heart in diastole, lungs pale, stomach 
contracted, a serous effusion in the thoracic cavity, and the 
subcutaneous tissue pale and oedematous. 
Novy has also obtained a poisonous proteid from cul- 
tures of this germ. The following experiments illustrate 
the effects obtained with this body : 100, 50, and 25 milli- 
grammes, respectively, were injected into three young rats 
from the same litter. The animal which received 100 mg. 
soon began to crawl about on its belly, being unable to rise. 
The eyes were soon filled with a thick secretion and the 
toes became red. Finally it became quiet, lying on its 
belly, with feet extended. The respirations became deeper, 144 
bacterial poisons. 
and a coma-like condition set in. The animal died, with- 
out convulsions, within about three hours. The rat which 
received 50 mg. went through the same course of symptoms, 
but these were less intense. Death resulted four hours 
after the injection. The one which received 25 mg. be- 
came very sick, but finally recovered, and one week later 
it was given another injection of 30 mg., which produced 
scarcely any effect. Then it was treated at intervals of 
five, three, five, two, and four days, respectively, to 40, 50, 
75, 100, and 125 mg. without effect. Three days after the 
last injection the animal was inoculated with one c. c. of a 
bouillon culture of the highly virulent germ. Only a 
slight temporary effect was observed during the first day, 
after which recovery was complete and permanent. A 
control rat which was given the same quantity of the cul- 
ture sickened the next day and died one week later. From 
this it will be seen that the animal was rendered immune 
against the disease. 
Schweinitz also reports the detection of a slightly 
poisonous base, which he designates as sueholotoxiu, and a 
poisonous proteid, and with these he has been able to secure 
immunity in guinea-pigs against the virulent germ. The 
proteid body is classed among the albumoses, and is said 
to crystallize in white, translucent plates when dried in 
vacuo over sulphuric acid and to form needle-like crystals 
with platinum chloride. No one else has reported a crys- 
talline bacterial proteid, and this body is deserving of a 
more extended study. 
Rabbit Septicaemia.—Hoffa has killed rabbits by 
inoculation with pure cultures of the bacillus of this disease, 
and has isolated from the bodies of these animals methyl- 
guanidin, while in the bodies of healthy rabbits this poison 
could not be found. The fatal dose of methylguanidin for 
rabbits was found to be 0.2 gramme when given subcu- 
taneously. Since Hueppe has suggested that the bacte- 
rium of chicken-cholera is identical with that of rabbit 
septicaemia, chickens were poisoned with methylguanidin, 145 
PUERPERAL fever. 
and the symptoms were observed to be analogous to those 
of the disease. 
Pneumonia.—Bonardi has made a chemical study of 
the diplococcus of Frankel. He finds certain poisons— 
ptomaines—which he has been unable as yet to obtain in 
quantity sufficient for ultimate analysis. Pie also claims 
to have secured immunity against the germ by treating 
rabbits with small quantities of the chemical poisons. 
Malignant (Edema.—Kerry fiuds that the bacillus 
of this disease decomposes albumin with the formation of 
fatty acids, leucin, hydro-paracumaric acid, and a foul- 
smelling oil of the composition C8Hlfi04. This oil is in- 
soluble in water, alkalies, and acids, easily soluble in ether, 
benzol, bisulphide of carbon, and alcohol. It is optically 
inactive, and on being oxidized furnishes valerianic acid. 
Nothing is said concerning its action upon animals. 
Among the gaseous products are carbonic acid, hydrogen, 
and marsh gas. The author was unable to determine 
whether or not free nitrogen is formed. 
Puerperal Fever.—Bourget claims to have isolated 
several ptomaines from the urine of women with puerperal 
fever. His conclusions are as follows: (1) In puerperal 
fever the urine contains highly poisonous bases. (2) The 
toxicity of the urine is most marked when the symptoms 
of the disease are most grave, and diminishes as the symp- 
toms abate. (3) The ptomaines obtained from the urine 
prove fatal when injected into frogs and guinea-pigs. (4) 
Toxic bases, resembling those obtained from the urine, were 
extracted from the viscera of a woman who had died of 
puerperal fever. CHAPTER VI. 
THE NATURE OF IMMUNITY-GIVING SUBSTANCES. 
Ogata and Jasuhara find that anthrax bacilli grown 
in the blood-serum of animals naturally immune to the 
disease will not on subsequent inoculation induce the dis- 
ease in animals naturally susceptible. Thus, anthrax 
germs grown in frog-blood make mice sick, but do not 
prove fatal to them, and those grown on the blood-serum 
of white rats or dogs have a similar effect upon rabbits; 
but germs grown in the blood of animals not immune kill 
both mice and rabbits. They also find that the injection 
of one drop of frog blood-serum or one-half drop of serum 
from a dog into a mouse, any time within seventy-two 
hours before to five hours after inoculation with anthrax, 
protects this animal from the disease. A guinea-pig weigh- 
ing 400 grammes was given twenty drops of frog’s blood 
diluted with the 0.6 per cent, salt solution and immediately 
thereafter inoculated with virulent anthrax; the animal 
became slightly sick, but soon recovered. The same was 
true of a rabbit weighing 1500 grammes which was treated 
with 8 c.c. of defibrinated dog’s blood. The experimenters 
conclude that one-fourth of a drop of the serum of the dog 
diluted to three times its volume with the salt solution is 
the smallest amount which will give immunity against 
anthrax to a mouse of 10 grammes. 
Kitasato and Behring have secured immunity in 
some animals against tetanus and diphtheria by the follow- 
ing methods: 
(1) By the method of Franker (for diphtheria), which 
has been given. (See page 128.) 
(2) By the addition of iodine trichloride to cultures four 
weeks old, in the proportion of 1 : 500 ; allow to stand for 
sixteen hours; inject 2 c.c. into the abdominal cavity of a 147 
IM M UNITY — GIVING SUBSTANCES. 
guinea-pig; three weeks later inject 0.2 c.c. of a culture 
in bouillon containing iodine trichloride in the proportion 
of 1 : 5500. 
(3) By the metabolic products of the diphtheria bacillus 
in the living body. In the pleural cavities of guinea-pigs 
killed by inoculation with the germ there is often a reddish, 
germ-free transudate; 10 c.c. to 15 c.c. of this kills guinea- 
pigs; small amounts give immunity. 
(4) By inoculating with the virulent germ and arresting 
the growth of the same with iodine trichloride, gold-sodium 
chloride, uaphthylamine, or carbolic acid. Of eight guinea- 
pigs, each of which was inoculated with 0.3 c.c. of a viru- 
lent culture, two, which were not treated, died within 
twenty-four hours; four, which had—two each—a 1 per 
cent, and a 2 per cent, solution of iodine trichloride injected 
immediately and at the place of inoculation, recovered ; of 
two which had the same treatment six hours after the 
inoculation, one died after four days. 
(5) By peroxide of hydrogen in diluted sulphuric acid. 
Guinea-pigs bear from 1 :4000 to } : 2500; mice, 1 : 2000 
to 1 : 800; rabbits, less than 1 : 1500 of this substance 
per body-weight. Injections of this solution before inocu- 
lation give more or less immunity, or, rather, increase the 
resistance to the disease; given after inoculation it hastens 
death. 
None of these methods are applicable to the prevention 
or treatment of the disease in man. 
Tizzoni and Cattani have reviewed the above state- 
ments in so far as they refer to tetanus. These experi- 
menters find that the addition of an equal volume of 
either a 2 per cent, solution of fresh chlorine water or 
iodine trichloride, or a 5 percent, solution of phenylic acid 
to the poisonous, filtered tetanus culture destroys the tox- 
icity of the same; but they state that the injection of these 
substances into animals either before or after inoculation 
with the germ has no effect upon the development or course 
of the disease. 
However, they do find that the blood-serum of an ani- 
mal which is immune will protect against either the 148 
BACTERIAL POISONS. 
living germ or tlie germ-free culture. Pigeons and dogs 
are but slightly susceptible to tetanus and they are made 
still less so by being treated for a number of times with 
small quantities of the virulent culture. After each re- 
covery these animals are found to be less susceptible, arid 
finally they acquire a high degree of immunity, and then 
their blood is employed in securing immunity in other 
animals much after the manner already detailed for 
anthrax. 
Tizzoni and Cattani have attempted to ascertain the 
nature of that constituent of the blood-serum which gives 
immunity. In these experiments serum from a dog which 
had been rendered immune against tetanus was employed. 
In the first place, a filtered culture of the tetanus germ 
was concentrated in vacuo at 40° until one-half c.c. of 
it would kill a rabbit within thirty-six hours. To this 
amount of the culture, the blood-serum was added after 
having been subjected to varied treatment, and the whole 
was subsequently injected into a rabbit. The blood-serum 
retains its antitoxic properties when kept in the dark at 
15° for some days, and it may be heated to 60° without 
injury. A temperature of 65° weakens, and one of 68° 
(tile temperature at which the serum coagulates) completely 
destroys the antitoxic properties of the serum. The 
“ tetanus-antitoxine ” is non-diffusible. It is precipitated 
from the blood-serum on the addition of absolute alcohol 
and from the dried percipitate it may be extracted either 
with water or glycerin, though very slowly with the 
latter. From these facts it is concluded that the antitoxin 
is a proteid with the characteristics of an enzyme. 
Hankin gives the following argument in favor of the 
theory that immunity is not due to ptomaines, but to pro- 
teids : “ It is generally admitted that in acquired immunity 
against a disease we are dealing (for the most part, at least) 
with a phenomenon of the nature of acquired tolerance of 
a poison. If we consider what this theory really implies, 
and, further, suppose that the poison involved is a pto- 
maine or other body of an alkaloidal nature, numerous 
diffculties immediately present themselves. For, iu the 149 
IMMUNITY-GIVING SUBSTANCES? 
first place, if acquired immunity is of this nature, we are 
dealing with an acquired tolerance of a poison, which 
tolerance is conferred by administering a single dose, or at 
most a very limited number of doses. Further, this ac- 
quired tolerance, thus easily obtained, is very permanent, 
lasting for mouths, or even years. Now, though acquired 
tolerance of alkaloids is constantly observed, it is but 
limited in degree, and only obtained as the result of a 
long-continued succession of doses.1 Secondly, since ac- 
quired tolerance of this hypothetical poison results in the 
microbe being no longer capable of living in the body, 
this theory implies that the poison in question is one that 
is produced by the microbe in order to live there. In 
other words, that it is a poison capable of lowering the 
bacteria-killiug power possessed by every living animal 
body.2 
“ Of course, it is conceivable that a ptomaine might be 
concerned in doing this, but, so far as I know, no parallel 
to such action can be found among bodies of an alkaloidal 
nature. 
“ When, however, we turn to what is known of poison- 
ous proteids, we at once find that they have properties 
analogous to those of the hypothetical immunity-giving 
poison. 
“ First, as regards the question of tolerance: Two poisons 
are known, which, in the nature of the tolerance they pro- 
duce, resemble the hypothetical poison in question. Both 
of them are albumoses. The first is the ordinary hemi- 
albumose of proteid digestion. It is known that the injec- 
tion of a single minute dose confers immunity against a 
1 Carbone claims to have obtained immunity in rabbits against the action 
of the proteus vulgaris by means of not more than two previous injections 
of small quantities of neurin obtained from cultures of the proteus. He 
still further states that immunity against the same germ is obtained by 
muscarin, which produces physiological effects practically identical with 
those of this neurin. 
2 With this statement we must take issue. The experiments already 
given in which immunity is induced in a susceptible animality the injection 
of the serum of the blood of an animal naturally immune show that the 
immunity-giving substance is not necessarily of bacterial origin, and cer- 
tainly that it is not necessarily a product of the germ against which the 
immunity is secured. 150 
BACTERIAL POISONS. 
further dose for a period of twelve hours. The second 
albumose is the poisonous principle of snake-poison. 
Sewall, in 1887, published a very interesting research on 
acquired immunity against snake-poison, lie showed that 
it was possible, by the injection of a few minute doses, to 
give pigeons such a tolerance of this substance that, three 
months after the treatment, they were able to stand what 
would otherwise be seven times the lethal dose. He sug- 
gests in his paper that, by inoculation with the ptomaines 
produced by bacteria, it may be possible to protect animals 
against their disease-producing powers, although the re- 
markable case of tolerance he had discovered suggested that 
not ptomaines, but albumoses, were the substances con- 
cerned in giving immunity against a disease;1 for I sug- 
gest that this fact—that the only cases of tolerance known 
which resemble the tolerance implied in disease-immunity 
are cases of tolerance against albumoses—strongly suggests 
that immunity against a disease is immunity against an 
albumose produced by the microbe.” 
In conformity with the above-stated theory, IIankin 
prepared, as we have already stated, from cultures of the 
anthrax bacillus a poisonous albumose, which, when em- 
ployed in small doses, gives immunity; in large doses, 
proves fatal. Hankin endeavored to separate any ferment 
that might be present and to which the immunity might 
possibly be due. A quantity of lime water was added to 
a solution of the albumose and the lime precipitated by the 
addition of phosphoric acid. Theoretically, the precipitate 
should contain any ferment present, and the immunity- 
giving property of the albumose would be diminished by 
the amount of the ferment thus removed, in case the im- 
munity be due to the ferment. However, the albumose 
was found to have lost none of its immunity-producing 
power. From this IIankin concludes that the albumose 
is the real immunity-producing agent. He does not in- 
1 We would suggest the faet that in 1887 it was not known that bacteria 
produce albumoses, and at that time the term “ptomaine” was employed 
to indicate all the bacterial poisons. development of infectious diseases. 
151 
form us whether or not any test of the phosphate precipi- 
tate was made. 
Bacterial Products which Favor the Develop- 
ment of Infectious Diseases.—Roger has made a 
very interesting contribution on this subject, and if his 
work be confirmed the question of mixed infection will 
become more important than it has been supposed to be. 
Rabbits are not naturally susceptible to the germ of char- 
bon symptomatique; indeed, inoculation with pure cultures 
of the bacillus has no visible effect. But Roger finds that 
if the staphylococcus pyogenes aureus, proteus vulgaris, or 
bacillus prodigiosus be injected into the animal at the same 
time with the germ of charbon symptomatique the latter 
develops and produces the disease. The same result is 
obtained when a sterilized culture of the bacillus prodigi- 
osus is employed. He at first supposed that the chemical 
products of the bacillus prodigiosus so lowered the vitality 
of the tissues that the pathogenic germ was enabled to 
establish itself; but he found that the same results were 
obtained when the two inoculations were made in distant 
parts of the body. The most marked effects were seen 
when the sterilized culture was injected into a vein and the 
charbon bacillus subcutaneously. In these instances the 
rabbits rapidly developed enormous tumors, and died 
within twenty-four hours. One drop of the sterilized cul- 
ture was found to be sufficient, when injected intravenously, 
to render rabbits susceptible to the pathogenic germ. 
In this connection it may be remarked that from time 
to time statements have been made which would lead us to 
infer that there are certain poisonous proteids which in 
some way yet unknown render the body especially suscep- 
tible to the invasion of bacteria. Rossbacii injected a 
poisonous albumose from the juice of the papain tree into 
the bloodvessels of animals and obtained a septicaemia. 
The blood was found to be filled with non-pathogenic 
germs which came from the intestines. The results of 
Rossbacii have, however, been questioned by others. 
Hankin makes the statement that a small dose of snake- 152 
BACTERIAL POISONS 
poison, which is too small to kill the animal outright, after 
a certain time may cause death from septicaemia. He says: 
“ The albumose of the snake-poison has apparently so far 
suppressed the germicidal power of the animal that ordinary 
decay-producing bacteria can increase and multiply in the 
blood. Further, it is often remarked that animals killed 
by a snake-bite putrefy rapidly, as if the bacteria-killing 
power of the blood-serum had been diminished.” A simi- 
lar statement has been made concerning the action of the 
poisonous albumose of jequirity seeds. Further investiga- 
tion must discover how much of truth and how little of 
error lie in these claims. CHAPTER VII. 
THE GERMICIDAL PROTEIDS OF THE BLOOD. 
As early as 1872 Lewis and Cunningham showed 
that bacteria injected into the circulation rapidly disappear. 
In the blood of twelve animals, which had been treated 
with such injections, bacteria could be found in only seven 
after six hours. In thirty animals, bacteria were found in 
the blood of only fourteen after twenty-four hours, and in 
seventeen animals, bacteria were found in only two when 
the examination was made from two to seven days after 
the injection. 
Iu 1874, Traube and Gscheidlen found that the 
blood taken from a rabbit into the jugular vein of which 
forty-eight hours before c.c. of a fluid rich in putre- 
factive germs was injected, remained without undergoing 
decomposition for months. These investigators attributed 
the germicidal properties of the blood to its ozonized 
oxygen. Similar results were obtained by Fodor and 
Wysokowicz. The latter accounted for the disappearance 
of the bacteria not by supposing that they were destroyed 
by the blood, but that they found lodgement in the capil- 
laries. 
The first experiments made with extra-vascular blood 
were conducted by Groiimann under the direction of A. 
Schmidt. It was found that anthrax bacilli, after being 
kept in coagulating plasma, were less virulent, as shown by 
their effects upon rabbits. Groiimann supposed that in 
some way the bacteria were influenced by the process of 
coagulation. 
Iu 1887, Fodor made a second’series of experiments in 
which he used blood taken from the heart, and showed the 
marked germicidal properties of this on anthrax bacilli. 
In 1888, Nijttall used defibriuated blood taken from 154 
BACTERIAL POISONS. 
various species of animals (rabbits, mice, pigeons, and 
sheep) and found that this blood destroyed the bacillus 
anthracis, bacillus subtilis, bacillus megaterium, and staphy- 
lococcus pyogenes aureus when brought in contact with 
them. Nissen continued this work and employed blood- 
serum as well as defibrinated blood. The conclusions 
reached were as follows : 
(1) The addition of small quantities of sterilized salt- 
solution or bouillon to the blood does not destroy its 
germicidal properties. 
(2) Cholera germs and Eberth’s bacilli are easily de- 
stroyed by fresh blood 
(3) For a given volume of blood there is a maximum 
amount of bacilli which can be added. If too many 
germs are used the destruction is incomplete. 
(4) Blood whose coagulability has been destroyed by 
the injection of peptone is still germicidal. 
(5) Filtered blood-plasma from the horse is germicidal. 
Bkhiiing has attributed the action of the blood of 
white rats on anthrax bacilli to the presence of a hypo- 
thetical basic body to which the decidedly alkaline reaction 
of the blood is supposed by him to be due. Later, he lays 
special stress upon the amount of carbonic acid gas in the 
blood-serum. 
Buchner has made a most exhaustive study of this 
subject, in which he has been aided by Voit, Sittmann, 
and Ortiienberger. The results of this work are stated 
as follows : 
(1) The germicidal action of the blood is not due to 
phagocytes, because it is not influenced by freezing and 
thawing the blood, by which the leucocytes of the blood 
of the rabbit are destroyed. 
(2) The germicidal properties of the cell-free serum must 
be due to soluble constituents. 
(3) Neither neutralization of the serum, nor the addition 
of pepsin, nor the removal of carbonic acid, nor treatment 
with oxygen have any effect upon the germicidal proper- 
ties of the blood. 
(4) Dialysis of the serum against water destroys its GERMICIDAL PROTEIDS OF THE BLOOD. 
155 
activity, while dialysis against 0.75 per cent, salt solution 
does not. In the diffusate there is no germicidal substance. 
The loss by dialysis with water must be due to the with- 
drawal of the inorganic salts of the serum. 
(5) The same is shown to be the case when the serum is 
diluted with water and when it is diluted with the salt 
solution. In the former the germicidal action is destroyed, 
while in the latter it is not. 
(6) The inorganic salts have in and of themselves no 
germicidal action. They are active only in so far as they 
affect the normal properties of the albuminates of the serum. 
The germicidal properties of the serum reside in its proteid 
constituents. 
(7) The difference in the effects of the active serum and 
that which has been heated to 55° is due to the altered 
condition of the albuminates. This difference may possibly 
be a chemical one (due to changes within the molecule), or it 
may be due to alterations in mycelial construction. The 
albuminates act on the bacteria only when the former are 
in an “active state.” 
Halliburton has prepared from the lymphatic glands 
a globulin which he designates as cell-globulin ,5, and which 
agrees with fibrin ferment in inducing coagulation in plasma. 
Hankin has tested the germicidal properties of this cell- 
globulin. His experiments have been conducted in the 
following manner: The lymphatic glands (in later experi- 
ments the spleen, also) of a dog or cat are freed as much 
as possible from fat and connective tissue, then finely 
divided, and extracted with a dilute solution of sodium 
sulphate (one part of a saturated sodium sulphate solution 
+ nine parts of water). The cell-globulin passes into 
solution, while the other proteids are but sparingly soluble. 
After twenty-four hours the fluid is filtered and mixed 
with an excess of alcohol. The voluminous precipitate 
containing the cell-globulin is collected on a filter aud 
washed with absolute alcohol. For use, a part is dissolved 
in water and a small quantity of a bouillon culture of the 
anthrax bacillus added. Plate cultures are made along 
with control plates from time to time, and in this way the 156 
bacterial poisons. 
germicidal property of the substance is demonstrated. 
Hankin closes this contribution with the following con- 
clusions : 
(1) Halliburton’s cell-globulin /? has marked germi- 
cidal properties. 
(2) In this respect it differs from fibrin ferment. 
(3) The germicidal property of this substance seems to 
be identical with that of serum as described by Buchner, 
Nissen, and Nuttall. 
(4) The active properties of the serum are probably due 
to this or to an allied body. 
In a more recent contribution Hankin designates the 
germicidal agents of the body as “defensive proteids.” 
He thinks it probable that blood-serum owes its activity 
to these bodies and that the assumption of an “active con- 
dition of the serum albuminate” made by Buchner is 
unnecessary. He also thinks that Behring’s supposed 
alkaline base exists in the form of an albumose. We know 
of three albumoses which are alkaline in reaction. These 
are the protomyosinose and deuteromyosinose of KI'jhne 
and Chittenden, prepared by the digestion of myosin, 
and the anthrax albumose of Martin. 
By a method similar to that which he had employed in 
the preceding experiments, Hankin has isolated a “defen- 
sive proteid ” from the blood-serum and the spleen of the 
rat. This substance belongs to the globulins, and the nat- 
ural immunity of the rat against anthrax is probably due 
to its existence in the blood. 
Stern finds that the blood taken from different men, or 
from the same man at different times, varies markedly in 
its germicidal properties; also, that the germicidal proper- 
ties of the blood when kept at 42° are at least as great as 
at the normal temperature of the body. These statements 
are substantially confirmed by Rovighl CHAPTER VIII. 
METHODS OF EXTRACTING PTOMAINES. 
From what has been given in the preceding pages, one 
may gather some idea of the peculiar difficulties with which 
the chemist has to contend in his endeavors to isolate the 
basic products of putrefaction. He has to deal with very 
complex substances, of the nature and reactions of many 
of which he must be ignorant. Besides, the substances 
which he seeks are often most prone to undergo decompo- 
sition, and in this way escape detection. Many ptomaines 
are volatile or decomposable at any temperature near that 
of boiling water. In these cases, solutions cannot be 
evaporated in the ordinary way and the poison separated 
from the residue. Indeed, the investigator has frequently 
been disappointed when on the evaporation of a solution, 
which he has demonstrated to be poisonous, he finds that 
the residue is wholly inert. Again, he may destroy the 
ptomaine by the action of reagents which he uses. So 
simple a procedure as the removal of a metallic base from 
a solution containing a ptomaine, by precipitation with 
hydrogen sulphide gas, has been known to destroy wholly 
the ptomaine. Probably the most perplexing difficulty in 
the isolation of these putrefactive alkaloids lies in the great 
number, complexity, and diversity of the other substances 
present in the decomposing mass. The same ptomaine may 
be present in equal quantities in two samples of milk, and 
yet it may be easily obtained from the one, while from the 
other only minute traces can be secured. The difference is 
due to the fact that the other constituents of the milk in 
the two samples are at different stages of the putrefactive 
process, and, consequently, differ greatly in their reactions 
and in their effects upon the agents employed to isolate 
the poison. All chemists will appreciate these difficulties. 158 
BACTERIAL POISONS. 
One of the first things tor the chemist who undertakes 
to do this work is to ascertain whether or not his reagents 
are pure. We have found a number of samples of German 
ether, which was imported on account of its supposed 
purity, to yield on spontaneous evaporation a residue which 
gave several of the alkaloidal reactions, and a few drops of 
which, injected under the skin of a frog, caused paralysis 
and death within a few minutes. We would advise that 500 
c.c. of the ether to be used should be allowed to evaporate 
spontaneously, and its residue, if there be one, be examined 
both chemically and physiologically. The basic substance 
which exists in some samples of sulphuric ether is pyridine. 
Guareschi and Mosso found commercial alcohol almost 
invariably to contain small quantities of an alkaloidal sub- 
stance, the odor of which is similar to that of nicotine and 
pyridine. Its solutions are precipitated by gold chloride, 
phosphowolframic acid, phosphomolybdic acid, potassium 
iodide, and Mayer’s reagent, but not by platinum chloride 
or tannic acid. It does not reduce, or reduces feebly, ferric 
salts. From one sample of alcohol they obtained a base 
which, in addition to the above reactions, did give a pre- 
cipitate with platinum chloride. Alcohol may be freed 
from these substances by distillation over tartaric acid. 
In amylic alcohol, Haitinger has found as much as 
0.5 per cent, of pyridine. It may be purified in the same 
manner as recommended for ethylic alcohol. 
Chloroform, when found to leave any residue on evapora- 
tion, should be washed first with distilled water, then with 
distilled water rendered alkaline with potassium carbonate, 
then dried over calcium chloride and distilled. 
Petroleum ether sometimes contains a base which has an 
odor similar to trimethylamine or pyridine, and which gives 
a precipitate with platinum chloride, forming in octahedra. 
Benzole may contain a similar substance. 
The following methods have been used for the purpose 
of extracting the putrefactive alkaloids : 
The Stas Otto Method.—This method depends upon 
the following facts : (1) The salts of the alkaloids are sol- METHODS OF EXTRACTING PTOMAINES. 
159 
uble in water and alcohol, and generally insoluble in ether, 
and (2) the free alkaloids are soluble in ether, and are re- 
moved from alkaline fluids by agitation with ether. These 
principles are capable of great variety in their application. 
The usual directions are as follows : Treat the mass under 
examination with about twice its weight of pure 90 per 
cent, alcohol, and from ten to thirty grains of tartaric or 
oxalic acid; digest the whole for some time at about 70°, 
and filter. Evaporate the filtrate at a temperature not ex- 
ceeding 35° either in a strong current of air or in vacuo 
over sulphuric acid. Take up the residue with absolute 
alcohol, filter, and again evaporate at a low temperature. 
Dissolve this residue in water, render alkaline with sodium 
bicarbonate, and agitate with ether. After separation re- 
move the ether with a pipette, or by means of a separator, 
and allow it to evaporate spontaneously. The residue may 
be further purified by redissolving in water and again ex- 
tracting with ether. 
The following modifications of this method are em- 
ployed : Instead of tartaric or oxalic acid, acetic acid is 
frequently used. 
When the fluid suspected of containing a ptomaine is 
already acid from the development of lactic or other organic 
acid, the addition of an acid is often dispensed with. 
Ether extracts are made from both acid and alkaline 
solutions. 
Chloroform, amylic alcohol, and benzine are used as sol- 
vents after extraction with ether. 
The modification of this method, as carried out by Selmi 
and Marlno-Zuco is given in detail as follows: 
The material is divided as minutely as possible, placed 
in a large flask, and treated with twice its volume of 90 per 
cent, alcohol, and acidulated with tartaric acid in the pro- 
portion of 0.5 gramme to 100 c.c. of the mixture, taking 
care from time to time that the reaction is permanently 
acid. The flask, which is connected with a reflux condenser, 
is now placed on the water-bath and kept at the constant 
temperature of 70° for twenty-four hours. While yet 
warm the liquid is transferred to a special apparatus for 160 
BACTERIAL POISOXS. 
filtration by the aid of atmospheric pressure. The liquid 
is poured upon a wet cloth supported upon a perforated 
porcelain funnel, which is connected below with a receiver 
exhausted by a water-pump or aspirator. In this way 
rapid filtration is secured, and by repeated washing the 
extraction is made thorough. The acid alcoholic liquid is 
now transferred to a special distillation apparatus. 
A large tubulated retort of ten litres capacity is con- 
nected by means of a cork to a large tubulated receiver. 
The tubulure of the retort is provided with a small per- 
forated cork, which carries a glass tube finely drawn out 
and extending to the bottom of the retort. The tubulure 
of the receiver is connected with Liebig’s bulbs containing 
dilute sulphuric acid (1 to 10), and the bulbs in turn are 
connected with a water-pump or aspirator. 
In order to prevent the passage of air through the corks, 
they are covered with animal membrane which has been 
freed from fat. By means of the aspirator a fine current 
of air is drawn through the liquid and suffices to keep it 
constantly agitated. The retort is kept on the water-bath 
at a temperature of from 28° to 30°. The receiver is kept 
cold by a current of water In this manner the distilla- 
tion of the alcohol goes on rapidly and conveniently. More- 
over, decomposition is so far prevented that volatile bases 
are never found in the bulbs. 
The aqueous residue, after the removal of the alcohol by 
distillation, is filtered and extracted with ether as long as 
anything is dissolved. It is then mixed with powdered 
glass and evaporated to dryness in vacuo. This residue is 
repeatedly extracted with absolute alcohol. The alcohol is 
distilled again in the apparatus already described. The 
residue is taken up with distilled wrater and filtered. It is 
then made alkaline with sodium bicarbonate and repeatedly 
extracted with ether, benzine, and chloroform. 
In order to obtain the base from the solvent, the greater 
part may be evaporated on the water-bath and the re- 
mainder allowed to evaporate spontaneously, or the re- 
mainder may be treated with dilute hydrochloric acid and 
the evaporation continued on the water-bath or in vacuo. brieger’s method. 
161 
Dragendorff’s Method.—The finely divided sub- 
stance is digested for some hours with water acidulated 
with sulphuric acid at from 40° to 50°. This is repeated 
two or three times, and the united filtered extracts are 
evaporated to a syrup. This is treated with four volumes 
of alcohol aud digested for twenty-four hours at 30.° After 
cooling, the alcoholic extract is filtered, the residue washed 
with 70 per cent, alcohol, and the united filtrates freed 
from alcohol by distillation. The aqueous residue, diluted 
if desirable, is filtered and submitted to the following ex- 
tractions : 
(1) The acid liquid is shaken with freshly rectified petro- 
leum ether as long as this reagent leaves any residue on 
evaporation. 
(2) The acid fluid is now extracted with benzine. 
(3) The next solvent used is chloroform. 
(4) The liquid is now again extracted with petroleum 
ether in order to remove traces of benzine and chloroform. 
(5) The liquid is now made alkaline with ammonia and 
successively extracted with petroleum ether, benzine, chloro- 
form, and amylic alcohol. 
(6) The remainder of the ammoniacal liquid is mixed 
with powdered glass, evaporated to dryness, the residue 
pulverized, aud extracted with chloroform. 
The residue obtained with each of the above solvents 
should be examined for ptomaines. 
Brieger’s Method.—The substance under examination 
is divided as finely as possible, and then heated with water 
slightly acidified with hydrochloric acid. During the 
heating care must be taken that the feebly acid reaction is 
maintained. The heating should continue for only a few 
minutes. The liquid is then filtered and concentrated, at 
first on a plate and then on the water-bath, to a syrup. If 
one has material which is highly odorous, as is the case 
frequently both with aqueous and alcoholic extracts of 
putrid material, Brieger recommends that a piece of 
apparatus devised by Bocklisch be used. The fluid to be 
evaporated is placed in a globular flask, the rubber stopper 162 
BACTERIA L POISONS. 
of which carries two small glass tubes. One of these, b, 
extends to the bottom of the flask, while A terminates just 
above the surface of the liquid. The tube, A, is connected 
with a water-pump or aspirator, which draws the vapor 
through the tube. In order to prevent the return of con- 
densed fluids, the end of A in the flask is curved upon 
itself. The tube, b, is finely drawn out and through it a 
current of air is constantly moving. This prevents the 
formation of a deposit or a pellicle in the fluid. By rcgu- 
lating the amount of air coming through this tube, more 
or less of a vacuum will be formed in the flask. After 
evaporation to a syrup, an extraction is made with 96 per 
cent, alcohol, and the filtered extract is treated with a warm 
alcoholic solution of lead acetate. The lead precipitate is 
removed by filtration, the filtrate evaporated to a syrup and 
again extracted with 96 per cent, alcohol. The alcohol is 
driven off; the residue taken up with water; traces of 
lead removed with hydrogen sulphide; and the filtrate, 
acidified with hydrochloric acid, evaporated to a syrup. 
This syrup is extracted with alcohol, and the filtrate pre- METHODS OF GAUTIER AND ETARD. 
163 
cipitated with an alcoholic solution of mercuric chloride. 
The mercury precipitate is boiled with water, and on ac- 
count of differences in solubility of the double compounds 
with mercury, one ptomaine may be separated from others 
at this stage of the process. (If thought best, the lead pre- 
cipitate may be freed from lead and carried through the 
following steps of the process. Brieger has found small 
amounts of ptomaines in the lead precipitate only in his 
work with poisonous mussels.) 
The mercury filtrate is freed from mercury, evaporated, 
and the excess of hydrochloric acid carefully neutralized 
with soda (the reaction is kept feebly acid), then it is again 
taken up with alcohol in order to free it from inorganic 
salts. The alcohol is evaporated, the residue taken up with 
water, the remaining traces of hydrochloric acid neutralized 
with soda; the whole acidified with nitric acid, and 
treated with phospbomolybdic acid. The phosphomolyb- 
date double compound is separated by filtration, and de- 
composed by neutral acetate of lead. This is hastened 
by heating on the water-bath. The lead is removed by 
hydrogen sulphide, the filtrate is evaporated to a syrup and 
taken up with alcohol, from which many ptomaines are 
deposited as chlorides, or double salts may be formed in 
the alcoholic solution. Brieger states that the chlorides 
as deposited from the alcoholic solution are seldom pure, 
and he advises for their purification, precipitation with 
gold chloride, platinum chloride, or picric acid, and, on 
account of differences in solubility of these double salts, 
the process of purification is rendered more easy. The 
chloride of the base is obtained by removing the metallic 
base with hydrogen sulphide; while the picrate is taken 
up with water, acidified with hydrochloric acid, and re- 
peatedly extracted with ether, in order to remove the 
picric acid. 
The Methods of Gautier and Etard.—The putrid 
matters, liquid and solid, are distilled at a low temperature 
in vacuo. The distillate (A) contains a considerable quan- 
tity of ammonium carbonate, some phenol, skatol, trimethyl- 164 
bacterial poisons. 
amine, and the volatile fatty acids. The residue after dis- 
tillation is treated in succession by ether and by alcohol. 
The extraction with ether (B) separates the ptomaines 
and some fatty acids. The alcoholic extract (C) removes 
the remainder of the fatty acids, as well as the acid and 
neutral nitrogenized bodies, almost all of which are crys- 
tallizable. The insoluble residue is boiled with dilute 
hydrochloric acid, with exclusion of air, finally evaporated 
to dryness, and the residue again extracted with alcohol. 
This new alcoholic solution (D) can be divided by acetate 
and subacetate of lead into two principal portions. 
By operating in this manner the complex products of 
putrefaction are readily separated into four portions. 
In his more recent work, Gautier has employed the 
following method : The putrid liquids, after the removal 
of fats, are feebly acidified with very dilute sulphuric acid, 
then distilled in vacuo at a low temperature. The distillate 
contains ammonia, phenol, iudol, and skatol. The syrupy 
residue, separated from any crystals which may have 
formed, is rendered alkaline with baryta, filtered, and ex- 
tracted a great number of times with chloroform, in order 
to dissolve the bases. The solution is distilled at a low 
temperature, either in vacuo or in a current of carbonic 
acid. The contents of the retort, on being treated with 
water and tartaric acid, separate into a brown resin and a 
liquid portion. The latter is removed and treated with a 
dilute solution of potash, when it gives off the odor of 
carbylamine, which was discovered by Gautier in 1866, 
and which, according to Calmer, is a constituent of the 
venom of toads. The alkali also sets free the bases, which 
are removed by extraction with ether, and the ether evapo- 
rated in a current of carbonic acid gas under slight pressure, 
then under a bell-jar over caustic potash. The bases may 
be separated by fractional precipitation with platinum 
chloride, or, if present in sufficient quantity, by distillation 
in vacuo. 
Still later, Gautier has modified his method as follows: 
The alkaline putrid liquid is treated with oxalic acid (in- 
stead of sulphuric acid) to free acidulation and as long as REMARKS UPON THE METHODS. 
165 
the fatty acids continue to separate. The liquid is then 
warmed and distilled as long as a turbid fluid passes over. 
Pyrrol, skatol, phenol, indol, volatile fatty acids, and some 
of the ammonia pass over. The portion which remains in 
the retort is rendered alkaline with lime-water. The pre- 
cipitate which forms, and which contains the greater part 
of the fixed fatty acids, is removed. The liquid portion, 
which is alkaline, is distilled to dryness, care being taken 
to receive the distillate in very dilute sulphuric acid. The 
bases and ammonia pass over. The distillate is neutralized 
(with sulphuric acid) and evaporated almost to dryness, 
then decanted from ammonium sulphate, which crystallizes. 
The mother-liquor is extracted with concentrated alcohol, 
which dissolves the sulphates of the ptomaines. After 
driving off* the alcohol, the residue is rendered alkaline 
with caustic soda, and successively extracted with ether, 
petroleum ether, and chloroform. 
The lime precipitate is dried and extracted with ether 
of thirty-six degrees, which removes any fixed bases that 
may be present. 
Remarks upon the Methods. — The fundamental 
difference between the Stas-Otto and the Dragendorff 
methods consists in the fact that in the former the first 
extraction is made with a dilute solution of an organic acid 
(tartaric usually), while in the second a similar solution of 
a mineral acid (sulphuric) is employed. In their various 
modified forms any solvent may be used for separating the 
alkaloid from the other constituents of the original solu- 
tion. Therefore, the question has been asked, Which is 
the more suitable acid for use in making the first solution ? 
The answer to this question will also be the one to the 
question, Which is the better method of extracting pto- 
maines, the Stas-Otto method or that of Dragendorff? 
The Italian chemists Guareschi and Mosso have at- 
tempted to answer this question experimentally, and the 
evidence which they have furnished is condemnatory of 
the method of Dragendorff. They show that basic 
bodies are formed by the action of the dilute sulphuric 16(3 
BACTERIAL poisons 
acid upon albuminous substances. As this point is of 
vital importance to the investigator in this branch of 
chemical science, we will give a brief abstract of the work 
of Guareschi and Mosso : 
One kilogramme of fresh meat was treated with dilute sul- 
phuric acid (in the proportion recommended in the Dea- 
gendorff method) and alcohol. The dark solution after 
filtration was made alkaline with ammonium hydrate and 
extracted with ether. The ethereal solution gave on evap- 
oration an oily substance which had the odor of extracts 
obtained from putrid fibrin. This substance, which was 
obtained in considerable quantity, was soluble in water and 
strongly alkaline in reaction. After neutralization with 
hydrochloric acid, its aqueous solutions gave the following 
alkaloidal tests: 
(1) With platinum chloride, a yellowish-red precipitate, 
insoluble in water, alcohol, and ether, and apparently iden- 
tical with the compound obtained from putrid fibrin with 
the same reagent. 
(2) With gold chloride, yellow precipitate, then reduc- 
tion to metallic gold. 
(3) With phosphomolybdic acid, a heavy, yellow precipi- 
tate, forming a blue solution on the addition of ammonium 
hydrate. 
(4) With phosphotungstic acid, a white precipitate. 
(5) With Mayer’s reagent, a heavy, whitish precipitate. 
(6) With picric acid, white precipitate, instantly. 
(7) With iodine in potassium iodide solution, a heavy 
kermes-red precipitate. 
(8) With tannic acid, white precipitate. 
(9) With mercuric chloride, white, amorphous precipi- 
tate. 
(10) With Marme’s reagent, heavy precipitate. 
(11) With potassium ferricyanide, no precipitate, but a 
cloudiness, with a formation of Prussian blue on the addi- 
tion of ferric chloride. 
The same quantity of this meat was also treated by the 
Stas-Otto method. The alcoholic extract was evaporated 
on the water-bath and not in vacuo. The acid was neu- REMARKS UPON THE METHODS. 
167 
tralized with sodium bicarbonate. The ether extract gave 
on evaporation a faintly yellow residue, of not unpleasant 
odor and feebly alkaline reaction. After neutralization 
with hydrochloric acid, it was only slightly soluble in 
water. The pale yellow filtrate gave no precipitate with 
Nos. 1, 2, 8, 9, and 10 of the above-mentioned reagents, 
but gave a slight turbidity with Nos. 3, 4, 5, 6, and 7, and 
with 11 formed Prussian blue. 
Guareschi and Mosso conclude from this and other 
experiments that the Dragendorff method is not suit- 
able for the extraction of ptomaines, and they recommend 
the employment of the Stas-Otto method with these con- 
ditions : (1) no more acid should be added than is abso- 
lutely necessary to keep the reaction acid ; (2) the heat 
used in evaporation should not be great, and it is better 
that evaporation should be made in vacuo. In this way, 
they say, no ptomaine will be obtained from fresh tissue. 
The same investigators extracted fresh flesh without the 
addition of any.acid. Thirty kilogrammes of perfectly fresh 
meat were digested for two hours at from 50° to 60° with 
about one and one-half volumes of water. The fluids of the 
meat contained enough acid to give to the whole of this solu- 
tion an acid reaction. It was evaporated to half its volume 
on the water-bath, filtered, and evaporated still further. 
The small residue was taken up with about four volumes 
of 96 per cent, alcohol. The reddish, alcoholic solution 
left on evaporation on the water-bath a brownish residue, 
which was dissolved in water and extracted with ether (A), 
then the solution was made alkaline with ammonium 
hydrate and again extracted with ether (B). 
A gave on evaporation and cooling crystals of methyl- 
hydantoin, while the mother-liquor contained acetic acid. 
B also yielded crystals of methyl-hydantoin, while the 
mother-liquor gave alkaloidal reactions with most of the 
general alkaloidal reagents, none with platinum chloride. 
Methyl-hydautoin does not give these reactions. 
Marino-Zuco has made many comparative tests with 
these two methods. He ascertained that by treating fresh 
eggs, brain, liver, spleen, kidney, lungs, heart, and blood 168 
BACTERIAL poisons. 
by either of the methods, he could obtain a substance which 
gave alkaloidal reactions, and which he demonstrated to be 
choline. His experiments led him to believe that choline 
did not exist pre-formed in these fresh tissues, but that it 
resulted from the action of the dilute acids upon lecithin. 
It was found most abundantly in those tissues which are 
rich in lecithin, such as the yolks of eggs, brain, liver, 
and blood; while only traces could be obtained from the 
whites of eggs, lungs, and heart. The method of Dragen- 
dorff was found to furnish much larger quantities of 
choline than could be obtained by the Stas-Otto method. 
Coppola agrees with his countrymen, mentioned above, 
in condemning the method of Dragendorff. 
Enough has been said to show that results obtained by 
the Stas-Otto method are much more reliable than those 
secured by the method of Dragendorff. However, the 
former is not a perfect method, nor has a perfect one yet 
been devised. The principal difficulties met with in the 
Stas-Otto method are as follows : 
(1) In most instances the extraction of the base is very 
incomplete. (2) The degree to which the putrefactive 
alkaloid is removed by the solvent will depend very 
largely upon the nature of the other substances present. 
This fact in some cases aids and in others hinders the 
labors of the investigator. Thus, several ptomaines, which 
when pure are wholly insoluble in ether, may be removed, 
in part at least, from organic mixtures by this solvent by 
passing into the solution along with other substances, but 
if the attempt is made to purify one of these bases by re- 
peated solution and extraction with ether, the result is a 
failure, because the more perfectly the alkaloid is freed 
from impurities, the less soluble it is in ether. This criti- 
cism, however, is equally applicable to the Dragendorff 
method, jand to all others in so far as extractions are made. 
However, we may state that whenever it is applicable 
this method is the best now employed. By it the sub- 
stances are submitted to the least chemical manipulation, 
and the results obtained are the most reliable. Many of 
the more complex putrefactive products are so easily de- 169 
REMARKS UPON THE METHODS 
composed or otherwise altered that the investigator should 
seek to isolate them by the simplest methods possible. If 
it can be done without the addition of any acid or without 
the application of heat, so much the better. 
Especially is the modification of this method employed 
by Marino-Zuco, and already described, to be commended. 
By his method, Brteger has discovered a considerable 
number of basic bodies and has given great impetus to the 
study of the chemistry of putrefaction. The method is 
capable of a great many modifications. As long ago as 1868, 
Bergmann and Sohmiedeberg employed precipitation 
with metallic salts in order to obtain sepsine from putrid 
yeast. The method used by them was as follows : Putrid 
yeast was diffused through parchment paper; the diffusate 
was acidified with hydrochloric acid, and treated with mer- 
curic chloride solution until a heavy cloudiness and, after 
some time, a slight precipitate formed. This was removed 
by filtration; the filtrate was rendered strongly alkaline 
with sodium carbonate, and then further treated with a 
solution of mercuric chloride as long as a precipitate 
formed. This precipitate was collected on a filter, washed, 
suspended in a little acidified water, and decomposed with 
hydrogen sulphide. The precipitate was removed, the free 
hydrochloric acid in the filtrate taken up with silver car- 
bonate, and the excess of silver removed with hydrogen 
sulphide. The filtrate was evaporated to dryness; the 
residue dissolved in alcohol (a part remaining insoluble), 
and acidified with sulphuric acid, when a colorless or 
slightly yellow crystalline precipitate formed. The crys- 
talline sepsine sulphate was purified by solution in water 
and precipitation with alcohol. 
Brieger has obtained some of his bases by a much sim- 
plified modification of his complete method, which we have 
given in full. For instance, in obtaining neuridine, he 
treated the aqudbus extract of the putrid material, after 
boiling and filtration, with mercuric chloride, collected the 
precipitate, decomposed it with hydrogen sulphide, evapor- 
ated the filtrate on the water-bath, and extracted the base 
from the residue with dilute alcohol. 170 
BACTERIAL POISONS. 
By this method and its modifications Brieger has 
obtained many brilliant results,among which maybe men- 
tioned his discovery of mytilotoxine, typhotoxine, and 
tetauine. However, the method is not free from criticism. 
The great number of chemical manipulations to which the 
organic matter is subjected is liable to lead to the formation 
of some basic substances and to the destruction of others. 
One is justified in considering the isolated base as pre- 
existing in the original material only when it produces 
symptoms identical with those caused by the substance from 
which it is extracted. There can be no doubt that by this 
method many ptomaines would be decomposed. With it 
Ehrenberg obtained from poisonous sausage only inert 
bases, and tyrotoxicon, the ptomaine of poisonous cheese, 
is decomposed both by heat and the hydrogen sulphide 
employed. The origin of the ptomaines possessing a mus- 
cariue-like action discovered by Brieger has been ques- 
tioned by Gram, who states tljat when the lactate of choline, 
an inert substance which is widely distributed both in plants 
and animals, is heated, it is converted into a poison with 
such an action. CHAPTER IX. 
METHODS OF ISOLATING THE DACTERIAL PROTEIDS. 
Hankin employed the following process in preparing 
his anthrax proteid : 
“The cultures are made in 0.1 per cent. Liebig’s extract 
of meat solution, to which some fibrin is added. The Lie- 
big’s extract is very difficult to sterilize, and must be heated 
for two or three hours in the steam sterilizer on two or 
three successive days. The fibrin must be added only after 
this has been done, and then the flask is re-sterilized by 
repeated heating to boiling-point, for a short time only on 
each occasion. If the fibrin were added at first it would 
be decomposed by the prolonged boiling. By the above 
method this only occurs to a slight degree, a mere trace of 
peptone being present in the sterilized culture-fluid. After 
sterilizing, this is inoculated with the blood of an animal 
dead of anthrax, and kept at the ordinary temperature. 
The anthrax forms a typical growth on the masses of fibrin, 
and samples of the liquid removed on successive days show 
a gradual increase in the strength of their biuret reaction. 
After about a week the liquid is filtered and the albumose 
extracted. The reason for not keeping the flask at a tem- 
perature of 37° is that the albumose is gradually decom- 
posed into peptone by the anthrax ferment present, and 
this change takes place more rapidly at the higher tempera- 
ture. For instance, I have found scarcely a trace of albu- 
mose in a culture which had been kept at 37° for a week, 
and which gave a strong biuret reaction. The albumose 
is separated from the culture-liquid thus prepared by satu- 
ration with ammonium sulphate. It is better to acidulate 
it slightly by adding a little acetic acid. The bulky pre- 
cipitate of albumose which then appears is filtered off, and 
the salt separated from it by dialysis. An excess of thymol 172 
BACTERIAL POISONS. 
must be added at this stage to prevent putrefaction, or the 
dialysis can be carried on in a current of water which is 
warmed to from 45° to 50° C., at which temperature the 
growth of microorganisms is inhibited. After dialyzing 
for twenty-four hours or more the greater part of the salt 
will have vanished, and the albumose will be found in 
solution in a considerable quantity of water which will not 
have passed through the parchment. It is now necessary 
merely to concentrate the solution and precipitate the 
albumose by the addition of alcohol. In my earlier ex- 
periments this was accomplished by evaporating in vacuo 
at a temperature of 45° to 48°. When at length the liquid 
has been reduced to a few cubic centimetres it is poured 
into alcohol, and the precipitated albumose is tiltered off, 
washed with the same reagent (alcohol), and dried. 
“ Evaporating in vacuo is a long and tedious process, 
and it requires a somewhat complicated apparatus. When 
it is used for pathogenic albumoses there is always a risk 
of the temperature employed destroying or diminishing 
their physiological properties. Further, if the albumose 
is allowed to evaporate to dryness, it may be difficult to 
make it pass into solution again. To avoid these difficul- 
ties I have designed a method of concentrating such solu- 
tions which is less objectionable. It depends on the prin- 
ciple that, if alcohol and water are placed on opposite sides 
of a membrane, the water rapidly dialyzes through to mix 
with the alcohol, while only traces of alcohol pass through 
to mix with the water. Consequently, if a watery solu- 
tion of albumoses is dialyzed against alcohol, the solution 
diminishes in bulk and is rapidly concentrated, owing to 
the passage of the water through the membrane. 
“My modus operandi is to place the dilute albumose 
solution in a parchment sausage skin which is immersed in 
a foot glass full of methylated spirit. The spirit can be 
changed after some hours if it is desired to prolong the 
process; but this is not usually necessary. In this way I 
have been able to bring 400 c.c. of albumose solution down 
to 100 c.c. in the course of a single night, at the ordinary 
temperature, without risk to the albumose or trouble to ISOLATING THE BACTERIAL PROTEIDS. 
173 
myself' The concentrated solution is then poured into 
absolute alcohol, which precipitates the albumose and re- 
moves any impurities that might be derived from the 
methylated spirit. This prolonged treatment with alcohol 
will tend to remove any free ptomaines or other substances 
soluble in alcohol. Peptones and salts present in the cul- 
ture liquid remained for the most part in solution when 
the albumose was precipitated with (NH4)2S04. No soluble 
proteids (except traces of peptone) were present in the cul- 
ture medium.” 
Ordinarily the bacterial proteids are isolated by preci- 
pitation with absolute alcohol, re-solution in water and re- 
precipitation with alcohol. However, as has been stated, 
Tizzoni and Cattani find that strong alcohol destroys the 
activity of the poison of their tetanus germ. The method 
employed in obtaining the bacterial cellular proteids has 
already been given (see page 130). CHAPTER X. 
THE IMPORTANCE OF PTOMAINES TO THE TOXICOLOGIST. 
The presence in the cadaver of substances which give 
not only the general alkaloidal reactions but respond to 
some of the tests which have hitherto been considered 
characteristic of individual vegetable alkaloids, must be 
of the greatest importance to toxicologists. The possi- 
bility of mistaking putrefactive for vegetable alkaloids 
should always be borne in mind by the chemist in making 
his medico-legal investigations. On the other hand, as we 
have seen in preceding chapters, cases of poisoning by 
ptomaines sometimes terminate fatally, and in such in- 
stances the chemist should not be satisfied with determin- 
ing the absence of mineral and vegetable poisons, but 
should strive to detect in the food or in the dead body 
positive evidence of the presence of the putrefactive 
alkaloid. 
We will give a brief account of those cases in which 
putrefactive substances have been found to resemble in 
their reactions the vegetable alkaloids. 
Coniine-like Substances.—The most celebrated case 
in which a substance giving reactions similar to those of 
coniine has been found, was the Braudes-Krebs trial, 
which took place in Braunschweig in 1874. From the 
undecomposed parts of the body two chemists obtained, 
in addition to arsenic, an alkaloid which they pronounced 
coniine. This substance was referred to Otto for further 
examination. Otto reported that the substance was 
neither coniine nor nicotine, nor any vegetable alkaloid 
with which he was acquainted. Otto converted the sub- 
stance into an oxalate, dissolved it in alcohol, evaporated 
the alcohol, dissolved the residue in water, rendered this CONIINE-LIKE SUBSTANCES. 
175 
solution alkaline with potash, then extracted the base with 
petroleum ether. On evaporation of the petroleum ether 
the alkaloid appeared as a bright yellow oil, which had a 
strong, unpleasant odor, quite different, however, from 
that of coniine. It was strongly alkaline and had an 
intensely bitter taste. At ordinary temperature it was 
volatile. From its aqueous solution it was precipitated 
by the chlorides of gold, platinum, and mercury. In 
these reactions it resembled nicotine, from which it differed 
in the double refractive and crystalline character of its 
hydrochloride. With an ethereal solution of iodine this 
substance did not give the Roussin test for nicotine, but 
instead of the long ruby-red crystals there appeared small, 
dark-green, needle-shaped crystals. 
This substance was found to be highly poisonous. Seven 
centigrammes injected subcutaneously into a large frog pro- 
duced instautaueous death, and forty-four milligrammes 
given to a pigeon caused a similar result. On account of its 
poisonous properties the jury of medical experts decided 
that the substance was a vegetable alkaloid. Otto says 
that this decision astounded the chemists. 
Buouakdel and Boutmy found in the body of a woman, 
who had died, after suffering, with ten other persons, from 
choleraic symptoms from eating of a stuffed goose, a base 
which gave the odor of coniine and the same reactions with 
gold chloride and iodine in potassium iodide, etc., as 
coniine. The same base was found in the remainder of 
the goose. But it did not give a red coloration with the 
vapor of hydrochloric acid, and it did not form butyric 
acid on oxidation, and although it was poisonous, it did 
not produce in frogs the symptoms of coniine poisoning. 
Selmi repeatedly found coniine-like substances in de- 
composing animal tissue. By distilling an alcoholic extract 
from a cadaver, acidifying the distillate with hydrochloric 
acid, evaporating, treating the residue with barium hydrate 
and ether, and allowing the ether to evaporate spontane- 
ously, he obtained a residue of volatile bases, the greater 
part of which consisted of trimethylamine. After remov- 
ing the trimethylamine, the residue had the odor of the 176 
BACTERIAL poisons. 
urine of the mouse. Later, Selmi obtained an unmistak- 
able coniine odor from a chloroform extract of the viscera 
of a person who had been buried six months, and in an- 
other ease ten months after burial. Two or three drops of 
an aqueous solution of the alkaline residue of the chloro- 
form extract allowed to evaporate on a glass plate gave off 
such a penetrating odor that Selmi was compelled to with- 
draw from close proximity to the substance. The odor 
imparted to his hands in testing the substance with the 
general alkaloidal reagents remained for half an hour. 
This volatile base seemed to be formed by the spontane- 
ous decomposition of other ptomaines. 
An aqueous solution of a ptomaine obtained by Selmi 
by extraction with ether according to the Stas-Otto 
method from the undecomposed parts of a cadaver had 
no marked odor, but after having been kept for a long time 
in a sealed tube it not only gave off a marked coniine 
odor, but the vapor turned red litmus-paper blue. Again, 
the sulphate of a ptomaine obtained from putrid egg- 
albumin, on standing formed in two layers, one of which 
was a golden-yellow liquid, which on being treated with 
barium hydrate gave off ammonia, and later, the odor of 
coniine. Since butyric and acetic acids were formed by 
the oxidation of this base, Selmi concluded that he had 
real coniine or methylconiine, and that it was formed by 
the oxidation of certain fixed ptomaines, or by the action 
of different amido bases on volatile fatty acids. There- 
fore Selmi believed in the spontaneous origin of coniine 
or closely allied bases in putrid matter, also in the exist- 
ence of a “ cadaveric coniine.” 
The substance which was found by Sonnexsgiiein in a 
criminal trial in East Prussia, and which was believed by 
that chemist to be the alkaloid of the water hemlock (cicuta 
virosa), is thought by Otto, Husemann, and others, to be 
a cadaveric coniine. Otto says that the symptoms re- 
ported in the case were not those of either coniine or 
cicuta. Sonxenscheix obtained the base six weeks after 
the exhuming of the body, which had been buried three 
months. The base had the odor of coniine, the taste of A NICOTINE-LIKE SUBSTANCE. 
177 
tobacco, gave with potassium bichromate and sulphuric 
acid the odor of butyric acid, and behaved witli reagents 
like coniine. 
Husemann states that at present it is very difficult, if 
not impossible, for the chemist to state with certainty that 
he has detected true coniine in the dead body. The symp- 
toms and the post-mortem appearances must conform with 
those induced by the vegetable alkaloid. The analysis 
must be made before decomposition sets in, and the amount 
of the base found must be sufficient for physiological ex- 
periments to be made with it. 
A Nicotine-like Substance.—Wolckenhaar ob- 
tained from the decomposed intestines of a woman, who 
had been dead six weeks, by extraction with ether from an 
alkaline solution, a base which bore a close resemblance to 
nicotine. The base was fluid, at first yellow, but on being 
exposed to the air, brownish-yellow. It was strongly alka- 
line in reaction and gave off an odor resembling nicotine, 
but stronger, not ethereal, but benumbing and similar to 
that of fresh poppy-heads. It was soluble in all propor- 
tions in water, and the solutions, which did not become 
cloudy on the application of heat, did not taste bitter, but 
were slightly pungent. The peculiar odor did not disap- 
pear on saturating the base with oxalic acid. The hydro- 
chloride was yellow, like varnish, had a strong odor, and 
became moist on exposure to the air. Under the micro- 
scope it showed no crystals, differing in this respect from 
nicotine hydrochloride. It differed from nicotine also in 
its reactions with potassio-bismuthic iodide, gold chloride, 
iodine solution, mercuric chloride, and platinum chloride. 
It also failed to give the Roussin test for nicotine. More- 
over, it could not be identified with trimethylamine, spar- 
teine, mercurialine, lobeline, or other fluid and volatile 
bases. 
The studies of Rorsch and Fassbender (page 28), of 
Schwanert (page 28), of Liebermann (page 30), and 
of Selmi (page 31), have already been referred to in a 
preceding chapter. 178 
bacterial poisons. 
Strychnine-like Substances.—In a criminal prose- 
cution at Verona, Ciotta obtained from the exhumed, but 
only slightly decomposed body, an alkaloid which gave a 
crystalline precipitate with iodine in hydriodic acid, a red 
coloration with hydriodic acid, and a color test similar to 
that of strychnine with sulphuric acid and potassium 
bichromate, and with other oxidizing agents. This sub- 
stance was strongly poisonous, but did not produce the 
tetanic convulsions which are characteristic of strychnine. 
Ciotta pronounced this substance as probably identical 
with strychnine. Portions of the body were subsequently 
submitted to Selmi for his opinion. Selmi found that 
the substance which gave the color-reaction was not crys- 
talline, and that there was only “the presumption of a 
bitter taste to it,” while one part of strychnine in 40,000 
parts of water is intensely bitter. Selmi also held that 
many ptomaines give reactions similar to strychnine with 
iodine in hydriodic acid and with hydriodic acid. He 
also held that its physiological properties were such that it 
could not be strychnine. This substance could hardly 
have been aspidospermine, which reacts with sulphuric 
acid and potassium bichromate similarly to strychnine, be- 
cause quebracho bark, in which this alkaloid is found, was 
not at that time used as a medicine or known in Italy. 
Ptomaines giving reactions similar to those of strych- 
nine, and also causing tetanic spasms, have been found in 
Italy in decomposed corn-meal. Selmi obtained one of 
these substances, but found that it differed from strychnine 
inasmuch as it could not be extracted with ether. 
Lombroso has named the poisonous substance found in 
decomposed corn-meal pellagroceine, but this is really a 
mixture of ptomaines, some of which produce narcosis and 
paralysis, and others produce the symptoms of nicotine 
poisoning instead of the spasms caused by strychnine. 
A Morphine-like Substance.—In the Sonzogua 
trial, at Cremona, Italy, the experts seem to have con- 
founded a ptomaine with morphine. This substance was 
not removed from either alkaline or acid solutions with 1)1 GIT A LINE—LIKE SUBSTANCES. 
179 
ether, but could be extracted with amylic alcohol. It 
reduces iodic acid, but in its other reactions, as well as in 
its physiological properties, it bore no resemblance to 
morphine. In frogs it arrested the heart in systole, which 
is said never to happen in poisoning with morphine. It 
failed to give both the ferric chloride and the Pellagri 
tests for morphine. 
In the same body there was found a substance which 
was extracted from alkaline solutions with ether, and which 
gave, with hydrochloric acid and a few drops of sulphuric 
acid, on the application of heat, a reddish residue similar 
to that obtained by the same reagents with codeine, but in 
its other reactions it did not resemble this alkaloid. 
Atropine-like Substances.—Many investigators have 
found products of putrefaction which in their mydriatic 
properties resemble atropine and hyoscyamine. To this 
class belongs the substance observed by Zuelzer and Son- 
NENSCHEIN. It wTas removed from alkaline solutions by 
ether, and formed microscopic crystals, an aqueous solution 
of which, when applied to the conjunctiva, produced a 
mydriatic effect, and, when administered internally, in- 
creased the action of the heart and arrested the movements 
of the intestines. Moreover, with certain alkaloidal re- 
agents, such as platinum chloride, it resembled atropine. 
But when heated with sulphuric acid and oxidizing agents 
it did not give the odor of blossoms (Reuss’s test). How- 
ever, Selmi found ptomatropines which with sulphuric 
acid and oxidizing agents did give the blossom odor as dis- 
tinctly as the vegetable atropine. These putrefactive bases 
also developed this odor spontaneously after standing for 
two or three days, and this does not happen with atropine. 
The odor was produced with the ptomatropines by nitric 
and sulphuric acids, both in the cold and on the applica- 
tion of heat, while these acids in the cold do not produce 
the odor with atropine. 
DreiTAline-like Substances.—Elsewhere we have 
referred to the discovery of a ptomaine belonging to this 180 
bacterial poisons. 
class by Borsch and Fassbender (see page 28). Trot- 
tarelli obtained a similar substance from the brain of a 
man in whose abdominal viscera he could find no poison. 
The sulphate of this base gave on evaporation an aromatic- 
smelling and astringent-ta ting residue. It became purple 
with sulphuric acid alone, and dark red with hydrochloric 
and sulphuric acids. On frogs this ptomaine showed no 
toxic effect. 
A Veiiateine-like Substance.—Brouardel and 
Boutmy obtained from a corpse which had lain in water 
for eighteen months, and a large portion of which had 
changed into adipocere, a ptomaine resembling veratrine. 
It was removed from alkaline solutions by ether. On 
being heated with sulphuric acid it became violet. With 
a mixture of sulphuric acid and barium peroxide it be- 
came, in the cold, brick-red; and, on being heated, violet. 
With boiling hydrochloric acid it took on a cherry-red 
coloration. However, it differed from veratrine, inasmuch 
as it reduced ferric salts instantly, and when injected into 
frogs subcutaneously it did not induce in them the spas- 
modic muscular contractions characteristic of veratrine. 
Bechamp obtained by the Stas-Otto method from the 
products of the pancreatic digestion of fibrin an alkaloid 
body which gave with sulphuric acid a beautiful carmine- 
red, similar to that given with veratrine. By digesting this 
substance with gastric juice, and again extracting, he 
obtained a body which behaved with sulphuric acid similar 
to curarine. 
A Delphinine-like Substance.—In 1870, General 
Gibbone, an Italian of prominence, died suddenly. His 
servant was accused of having poisoned him. Two chem- 
ists of some reputation reported the presence of delphinine 
in the viscera. It seemed somewhat improbable that the 
servant should know anything of so rare a substance, or 
that he should have been able to obtain it. However, two 
or more varieties of staphisagria grow in Southern Italy, 
and it was possible that the servant had used some prepara- A COLCHICINE-LIKE SUBSTANCE. 
181 
tion made by himself from the plant. The supposed alka- 
loid was given to Selmi, of Bologna, for further study. It 
was removed from alkaline solutions by ether. When 
heated with phosphoric acid it became red, and when 
brought in contact with concentrated sulphuric acid, red- 
dish-brown. In these tests the substance resembled delph- 
inine, but with sulphuric acid and bromine water, also 
with Froiide’s reagent, the colorations characteristic of 
the vegetable alkaloid failed to appear. Moreover, Selmi 
showed that delphinine gave the following reactions, to 
which the suspected substance did not respond : (1) Delph- 
inine dissolved in ether, and treated with a freshly prepared 
ethereal solution of platinic chloride, gives a white, floccu- 
lent precipitate which is insoluble in an equal volume of 
absolute alcohol. (2) Delphinine gives precipitates with 
auro-sodium hyposulphite, and with a sulphuric acid solu- 
tion of cupro-sodium hyposulphite, the latter precipitate 
being soluble in an excess of the reagent. 
Finally, Oiaccia and Vella showed that while delph- 
inine arrests the heart of the frog in diastole, the suspected 
substance arrests it in systole. 
A Colchicine-like Substance.—Baumert found in 
a suspected ease of poisoning, twenty-two months after 
death, a substance which gave many of the reactions for 
colchicine. It was extracted from acid solutions with ether, 
to which it imparted a yellow color. On evaporation of 
the ether a yellow, amorphous substance remained, and 
this dissolved in warm water with yellow coloration. It 
could be extracted from acid solutions also by chloroform, 
benzol, and amylic alcohol, but not by petroleum ether. It 
was removed with much more- difficulty from alkaline 
solutions. 
All the extracts were yellow, and left on evaporation a 
feebly alkaline, markedly bitter, sharp-tasting, amorphous, 
yellow residue, which dissolved in water and dilute acids 
incompletely, forming a resin. When this resin was dis- 
solved in dilute sodium hydrate, and the solution rendered 182 
BACTERIAL POISONS. 
acid by sulphuric acid, the same reactions were obtained as 
with the original extract. 
With phosphomolybdic acid, phosphotuugstic acid, potas- 
sio-bismuthic iodide, potassio-mercuric iodide, iodine in 
potassium iodide, tannic acid and gold chloride, this sub- 
stance gave the same reactions which were obtained by 
parallel experiments with genuine colchicine; thus, the 
tannic acid precipitates were both soluble in alcohol, and 
the precipitates with phosphomolybdic acid in both cases 
became blue on the addition of ammonium hydrate. 
Concentrated sulphuric and dilute nitric and hydrochloric 
acids dissolved the supposed colchicine with yellow colora- 
tion. Strong nitric acid (1.4 sp. gr.) colored the substance 
dirty red, scarcely to be called a violet. When the sub- 
stance was purified as much as possible, this color became 
a beautiful carmine-red. The addition of water changed 
the red into yellow, and caustic soda produced a dark, dirty 
orange. 
In general, in the above-mentioned reactions, the putre- 
factive product agreed with the real colchicine, but the 
former gave precipitates with picric acid and platinum 
chloride, while the latter gives no precipitates with these 
reagents. 
In 1886, Zeisel proposed the following test for colchi- 
cine : When a hydrochloric acid solution of the alkaloid is 
boiled with ferric chloride, it becomes green, sometimes 
dark-green and cloudy. Now, if the fluid be agitated with 
chloroform, the chloroform will sink, taking up the coloring 
matter, and appearing brownish, granite-red or dark, and 
the supernatant fluid clears up without becoming wholly 
colorless. 
Baumert applied this test to both colchicine and the 
putrefactive product. To from two to five cubic centi- 
metres of the suspected solution in a test-tube, he added 
from five to ten drops of strong hydrochloric acid and 
from four to six drops of a ten per cent, solution of ferric 
chloride, then heated the mixture directly over a small 
flame until it was evaporated to half its volume or less. In 
the presence of one milligramme of colchicine the originally MORPHINE. 
183 
bright-yellow solution became gradually olive-green, and, 
on further concentration, dark-green aud cloudy. Then, 
on shaking the fluid with chloroform, admitting as much 
air as possible, the chloroform subsided, having a ruby-red 
color if as much as two milligrammes of colchicine were 
present, and a bright yellow if only one milligramme, and the 
supernatant fluid became of a beautiful olive-green. When 
ether, petroleum ether, benzol, carbon disulphide, or arnylic 
alcohol was substituted for the chloroform, the coloration 
did not appear. From this Baumert infers that the red 
coloring matter is either only soluble in chloroform, or that 
it is not formed until the chloroform is added. 
Baumert found this test of great value in decidiug 
whether or not the substance which he found was colchi- 
cine. The putrefactive product did not respond to the 
test. 
Some of this substance was sent to Brieger, who de- 
cided that it was not a base, but a peptone-like substance. 
It was also found to be inert physiologically. 
Before these investigations were made by Baumert, 
Liebermann had found the same or a similar colchicine- 
like substance in the cadaver. His description differed 
from that of Baumert only in regard to the taste of the 
substance, Liebermann having failed to observe any 
marked taste iu the substance which he found, while, as has 
been stated, Baumert reported a distinctly bitter taste. 
A colchicine-like substance has been found in beer, and 
it has been suggested that it was this which the above- 
mentioned toxicologists found in the bodies which they 
examined, but Liebermann states that the man whose 
body he examined had been a total abstainer from beer. 
Tamba compared the reactions of ptomaines obtained 
from putrid sausage with similar reactions of various alka- 
loids, and then ascertained the effect upon the alkaloidal 
reactions by mixing the alkaloids with the ptomaines. His 
results are as follows : 
Morphine.—Ptomaines are colored yellow with nitric 
acid; reddish-yellow with concentrated sulphuric acid; 184 
BACTERIAL POISONS. 
blue, violet, then green with Frohde’s reagent; yellow 
when evaporated with concentrated sulphuric acid, then 
treated with hydrochloric acid aud decomposed with sodium 
bicarbonate. The ptomaines reduce ferric chloride, but not 
iodic acid. With sugar and concentrated sulphuric acid, 
they give a yellow coloration. 
Mixtures of the ptomaines and morphine give absolutely 
characteristic reactions for morphine with sugar and sul- 
phuric acid, the violet coloration appearing distinctly; aud 
by evaporation on the water-bath with sulphuric acid, addi- 
tion of hydrochloric acid and decomposition with sodium 
bicarbonate, the violet color appearing. Iodic acid is re- 
duced by morphine in the presence of ptomaines, only 
when the ptomaines are present in minute quantity. 
The other reactions for morphine are not applicable in 
the presence of ptomaines. 
Strychnine.—The characteristic color reaction for this 
alkaloid, with potassium bichromate and sulphuric acid, is 
not atfected by the presence of ptomaines.1 
Brucine.—The nitric acid reaction for brucine is not 
affected by ptomaines. On the other hand, the reaction 
with sulphuric and nitric acids, in which a red coloration 
is obtained, is scarcely visible in the presence of ptomaines. 
The action of mercuric nitrate and heat on brucine, by which 
a violet coloration is produced, is not destroyed bv the 
presence of ptomaines. 
Veratrine.—The characteristic coloration of veratrine 
by concentrated sulphuric acid is not influenced by pto- 
maines. The same is true of the cherry-red coloration with 
concentrated hydrochloric acid. On the contrary, the action 
of sugar and sulphuric acid on veratrine is without result 
in the presence of ptomaines. 
Atropine.—The deep violet coloration produced by 
fuming nitric acid, subsequent concentration, and the addi- 
1 In contradiction to this, see page 178. DELPHININE. 
185 
tion of alcoliolic potassium hydrate, is not affected by the 
presence of ptomaines. On the other hand, the character- 
istic odor produced by the action of sulphuric acid and 
heat on atropine is scarcely recognizable when ptomaines 
are present. 
Narceine.—The blood-red color produced by concen- 
trated sulphuric acid fails in the presence of ptomaines. 
Colchicine.—Fuming nitric acid colors the ptomaines 
reddish-yellow, but the violet coloration of colchicine with 
nitric acid appears in well-defined form, even in the pres- 
ence of ptomaines. The other reactions for colchicine are 
valueless when ptomaines are present. 
Codeine.—The blue coloration of codeine with concen- 
trated sulphuric acid holds good when ptomaines arc present. 
The same is true of the reaction with sulphuric acid, heat, 
and the subsequent addition of nitric acid. Froiide’s re- 
agent fails with codeine when mixed with ptomaines, inas- 
much as the bluish coloration rapidly passes into a brown. 
Aconitine.—Phosphoric acid and concentrated sulphuric 
acid are without reaction on the alkaloid when mixed with 
ptomaines. 
Picrotoxine.—The reducing action of picrotoxine on 
alkaline copper sulphate solution is seriously affected by the 
presence of ptomaines. The same is true of other tests for 
this poison. 
Delphinine.—The reaction of delphinine with sulphuric 
acid and bromine water, as well as the one with Frohde’s 
reagent, is so much influenced by the presence of ptomaines 
that the alkaloid cannot be recognized. 
These results are to be accepted with caution, as it is not 
reasonable to suppose that all ptomaines will affect the test 
for the vegetable alkaloids in the same manner or to the 186 
bacterial poisons 
same degree. Moreover, there is no proof that Tamba 
worked with pure ptomaines. 
Tamba has also proposed to separate vegetable from 
putrefactive alkaloids by adding to ethereal solutions of 
mixtures an equal volume of a saturated ethereal solution 
of oxalic acid, and allowing to stand, when the oxalates of 
the vegetable alkaloids will separate in crystalline form, 
and the oxalates of the ptomaines will remain in solution. 
In other words, the oxalates of the vegetable alkaloids are 
insoluble in ether, while the oxalates of the putrefactive 
alkaloids are soluble in ether. But, in contradiction to this, 
Bocklisch states that the oxalate of cadaverine is insoluble 
in ether. 
The most important work which the toxicologist is called 
upon to do at present is to isolate and identify beyond all 
question the bacterial poisons. This work has become im- 
portant on account of the frequent occurrence of poisoning 
from articles of infected food. CHAPTER XI. 
CHEMISTRY OF THE PTOMAINES. 
The basic substances described in the following pages 
are arranged, so far as possible, in the regular natural order. 
An inspection of the list of these bases will show the remark- 
able fact of the predominancy of the amine type. Almost 
two-thirds of the known ptomaines contain only C, H, and 
N, and represent simple ammonia substitution compounds. 
Of the oxygenated bases, all of those whose constitution is 
known possess the trimethylamine molecule as their basic 
constituent, and it is quite probable that most, if not all, of 
the remaining ptomaines will be found to possess the same 
or a similar basic nucleus. 
It will be seen, furthermore, that a very large number of 
the ptomaines described possess little or no toxic action, 
and are, therefore, physiologically inert. It would seem, 
as Brieger has already pointed out, that a certain quantity 
of oxygen is necessary to the formation of poisonous bases. 
A free supply of oxygen, on the other hand, invariably 
yields non-toxic ptomaines. The poisonous bases begin to 
appear on about the seventh day of putrefaction, and in 
turn disappear if this is allowed to go on for a considerable 
period of time. 
Methylamine, CH3.NH2.—This is the simplest organic 
base that is formed in the process of putrefaction. It is 
ammonia in which one atom of hydrogen has been replaced 
by the methyl radical. It occurs in herring-brine (Tol- 
lens, 1866 ; Bocklisch, 1885) ; in decomposing herring, 
twelve days in spring (Bocklisch) ; in pike, six days in 
summer (Bocklisch) ; in haddock, two months at a low 
temperature (Bocklisch) ; in the fermentation of choline 
chloride (Hasebroek). Brieger has shown it to be 188 
BACTERIAL POISONS. 
present in cultures of comma bacillus on beef-broth which 
were kept for six weeks at 37°-38°. Ehrenberg re- 
ported its possible presence in poisonous sausage, and ob- 
tained it by growing a bacillus from this source on intes- 
tines (1887). In Brieger’s method, methylamine is found 
both in the mercuric chloride precipitate and filtrate. The 
mercury double salt is readily soluble in water, and can 
thus be separated from any accompanying cadaverine or 
putrescine. Methylamine is an inflammable gas of strong 
ammoniacal odor, and burning with a yellow flame. It is 
readily soluble in water, and its solutions give reactions 
similar to those of ammonia. Its salts are, as a rule, also 
soluble in both water and alcohol. 
The Hydrochloride, CH3 NII2.HC1, crystallizes in 
large deliquescent plates. On being heated with alkali, it 
gives off the odor of methylamine. 
The Platinochloride, (CH3.NH2.HCl)2PtCl4 (Pt = 
41.31 per cent.),1 yields hexagonal plates which usually 
occur heaped up in several layers. It is soluble in about 
fifty parts of water at ordinary temperature, and can be 
readily recrystallized from hot water. It is insoluble in 
absolute alcohol and in ether. 
The Aurochloride, CH3.NH2.HC1.AuC13 + H20, 
forms prisms, which are readily soluble in water. There 
is also a readily soluble picrate. 
Methylamine does not possess any toxic action, even when 
given in fairly large doses. This physiological indifference 
is shared by nearly all the monamines and diamines that 
have been obtained among the products of putrefaction. 
Dimethylamine, (CH3)2.NH, has been found in putre- 
fying gelatin, ten days at 35° (Brieger, 1885); in yeast 
decomposing in covered vessels for four weeks during sum- 
mer (Brieger); in decomposing perch, six days in summer 
(Bocklisch); and in herring-brine (Bocklisch, 1886). It 
lias been found in poisonous sausage, and in cultures of a 
1 The percentages given in the following pages are calculated from Au= 
196.64 (Kriiss), Pt = 194.46 (Seubert), Cl = 35.37, 0 = 15.96. CHEMISTRY OF THE PTOMAINES. 
189 
bacillus obtained from this source, on liver and intestines 
(Ehrenrerg, 1887). It is also formed, together with 
trimethylamine, when neuridine hydrochloride is distilled 
with sodium hydrate (Brieger, I., 23). It occurs in the 
mercuric chloride precipitate as well as filtrate. From 
cadaverine it can be separated by platinum chloride, since 
cadaverine platinochloride is difficultly soluble in cold water, 
and recrystallizes from hot water, whereas the dimethyl- 
amine double salt remains in the mother-liquor. In like 
manner it can be separated from neuridine. From choline 
it can be isolated by recrystallizing the mercuric chloride 
precipitate from hot water. 
The free base is a gas at ordinary temperature, but can 
be condensed to a liquid which boils at 8° -9°. The 
hydrochloride, (CH3)2.NIT.HC1, crystallizes in needles, 
which deliquesce on exposure to air and are soluble in abso- 
lute alcohol (Brieger, I , 56). It is insoluble in absolute 
alcohol (Bocklisch) but soluble in chloroform (Behrend), 
and can then be separated from methylamine hydrochloride, 
which is insoluble in chloroform. 
The Platinochloride, [(CII3)2.NH.HCl]2PtCl4, (Pt 
= 39.00 per cent.), crystallizes in long needles, which are 
easily soluble in hot water, less soluble in cold water. Some- 
times it forms orange-yellow plates or prisms, or else small 
needles. 
The Aurochloride, (CH3)2.NH.HC1.AuC13, forms 
needles (Bocklisch), or large yellow monoclinic plates 
(Hjortdahl), which are insoluble in absolute alcohol. 
Trimethylamine, C3H9N = (CH3)3N, has been known 
for a long time to occur in animal and vegetable tissues. 
Dessaignes showed its presence in leaves of Chenopodium 
(1851), in the blood of calves (1857), and later in human 
urine. It has been obtained from ergot (Seeale cornutum) 
by Walz (1852) and Brieger (1886); from herring-brine 
by Wertheim, Winkles, Tollens, and Bocklisch. 
In these substances, with the exception of herring-brine, it 
probably does not exist pre-formed, but is rather a product 
of the method employed for its isolation. In fact, Brieger 190 
bacterial poisons. 
has shown that it does not exist in ergot, but is formed at 
the expense of the choline present, which, on distillation 
with potash, decomposes and yields trimethylamine and 
glycol. Thus: 
C2H4OH.N(CII3)3.OH = N(CII3)3 + C2H4(OH)2. 
It is also formed when betaine and neuridine are distilled 
with potash. It may have a similar origin in most of the 
other cases, since choline is now known to be widely dis- 
seminated in plants and animals, either as such or as a 
constituent of the more complex lecithin. Trimethylamine 
has been found in the putrefaction of yeast (IIesse, 1857 ; 
Muller, 1858); in cheese after six weeks in midsummer 
(Brieger) ; in human liver and spleen after from two to 
seven days (Brieger) ; in perch after six days in mid- 
summer (Bocklisch); in mussel (Mytilus edulis) after six- 
teen days (Brieger) ; in putrefying brains after from one 
to two months, and in fresh brains (Guareschi and 
Mosso); in cultures of the Streptococcus pyogenes on beef- 
broth, bouillon, meat extract, and blood-serum, and from 
cultures of the comma bacillus (Brieger). It has also 
been found in cod-liver oil. Ehrenberg (1887) reports 
its presence in considerable quantity in poisonous sausage, 
and in cultures of a bacillus, isolated from this, grown on 
liver, intestines, and meat bouillon. 
Trimethylamine is found both in the mercuric chloride 
precipitate and filtrate. It remains in the mother-liquor 
from which cadaverine, neuridine, and dimcthylamine pla- 
tinochlorides have crystallized. If an aqueous solution of 
mercuric chloride is used as the precipitant, the trimethyl- 
amine will be found almost entirely in the filtrate, from 
which it can be obtained after removal of the mercury by 
evaporating the filtrate to dryness, extracting with alcohol, 
and treating the solution thus obtained with alcoholic pla- 
tinum chloride. 
The free base is a liquid possessing a strong, fish-like 
odor. Its boiling-point is 9.3°. It is strongly alkaline in 
reaction and freely soluble in water. 
The Hydrochloride, (CH3)3N.HC1, is deliquescent and chemistry of the ptomaines. 
191 
freely soluble in water and alcohol. Heated to 285° it 
decomposes. With alkalies it gives off the odor of the 
free base. 
The Platinochloride, [(CH3)3N.HCl]2PtCl4 (Pt = 
36.92 per cent.), is soluble in hot water, from which, on 
cooling, it recrystallizes in orange-red octahedra or needles, 
which do not lose water when heated at 100°-110° (Bock- 
lisch). 
The Aurochloride, (CH3)3N.HC1.AuC13 (An = 49.39 
per cent.), is easily soluble, and hence can be separated from 
choline aurochloride, which is difficultly soluble. Similarly, 
this base can be separated from ammonia by the use of gold 
chloride. 
Trimethylamine is not a strong poison, since very large 
doses of it must be given in order to bring out any physio- 
logical disturbances. 
Kthylamine, C2H5.NH2, is formed in putrefying yeast 
(Hesse, 1857); in wheat flour (Sullivan, 1858); and also 
in the distillation of beet-sugar residues. 
It is a strongly annnoniacal liquid boiling at 18.7°, and 
is miscible with water in every proportion. Like the other 
amines, it is combustible. It possesses strong basic prop- 
erties, and is capable of expelling ammonia from its salts in 
a manner analogous to the action of the fixed alkalies. 
The Hydrochloride, C2H5.HH2.HC1, forms deliques- 
cent plates, which melt at 76°-80°. It is readily soluble 
in water and alcohol. 
The Platinochloride, (C2H5.NH2.HCl)2PtCl4, forms 
orange-yellow rhombohedra (Weltzien), or hexagonal- 
rhombohedral crystals (Topsoe). 
The Aurochloride, C2H5.NH2.HC1.AuC13, forms gold- 
yellow monoclinic prisms, readily soluble in water. 
With picric acid it forms short brown prisms, not very 
soluble in water. 
Diethylamine, C4HuN = (C2II5)2NH, has been ob- 
tained by Bocklisch from pike which were allowed to 
putrefy for six days in summer ; and by growing a bacillus 192 
bacterial poisons. 
obtained from poisonous sausage on intestines and on meat 
bouillon (Ehrenberg, 1887). 
It is an inflammable liquid which boils at 57.5°, possesses 
strong basic properties, and is soluble in water. 
The Hydrochloride, (C2H5)2NH.HC1, crystallizes in 
needles (Bocklisch) ; in long needles and prisms from 
absolute alcohol; in plates from ether-alcohol. These are 
not deliquescent and are easily soluble in water and in 
chloroform ; rather difficultly in absolute alcohol. Heated 
with sodium hydrate it gives off alkaline vapors. From an 
alcoholic solution it is precipitated by addition of alcoholic 
mercuric chloride. The mercury double salt is difficultly 
soluble in hot water, from which it recrystallizes on cooling. 
The Platinochloride, [(C2H3)2.lS[H.HCl]2PtO]4, crys- 
tallizes in orange-yellow monoclinic crystals, which are 
easily soluble in water. 
The Aurochloride, (C2H5)2.NH.HC1.AuC13 (An = 
47.71 per cent.), forms trimetric crystals (Topsoe), which 
are difficultly soluble (Bocklisch). It melts at about 165°. 
With picric acid it forms an easily soluble picrate (Lea). 
Triethylamine, C6H15N = (C2H5)3N, was obtained 
by Brieger (1885) from haddock which were exposed for 
five days in an open vessel during summer. He obtained 
it by distilling with potash, after removal of platinum by 
hydrogen sulphide, the mother liquor from which neuridine, 
the base C2H8N2, muscarine, and gadinine had successively 
crystallized (see Gadinine). It has also been found by 
Bocklisch (1886) in putrid pike, and by Ehrenberg 
(1887). The latter obtained it from cultures of a bacillus, 
found in poisonous sausage, and grown on meat bouillon. 
The free base is oily in character and possesses an am- 
moniaeal odor. It is but slightly soluble in water, and 
boils at 89°-89.5°. 
The Platinochloride, [(C2lI5)3N’.HCl]2I)tCl4 (Pt = 
31.84 per cent.), crystallizes in needles which are readily 
soluble in water. 
With mercuric chloride the aqueous solution gives no 
precipitate. chemistry of the ptomaines. 
193 
With picric acid it yields yellow needles which are but 
slightly soluble in cold water. 
Propylamine, C3II7.jNtH2, is isomeric with trimethyl- 
amine, and can therefore be easily confounded with that 
base. There are two propylamines possible represented 
by the formulae CH3.CH2.CH2.NH2 and ((1H3)2.CH.NIf2. 
The former, or the normal compound, boils at 47°-48°, 
whilst the latter, or iso-propylamine, boils at 31.5°. Both 
are liquids possessing an ammoniacal, fish-like odor. They 
form crystalline salts ; the hydrochlorides melt respectively 
at 155°-158°, and at 139.5°. 
Iso-propylamine (?) has been found among the distilla- 
tion products of the vinasse of beet-root molasses. Propyl- 
amine has been obtained by Briefer (1887) from cultures 
of the bacteria of human feces on gelatin. Schwanert 
has isolated from the organs of a cadaver a basic substance 
which was said to possess an odor similar to propylamine. 
Butylamine, C4HuN, was obtained by Gautier and 
Mourgues (1888) in cod-liver oil. It forms a colorless, 
mobile, alkaline liquid, the boiling-point of which they 
found to be 86° at 760 mm. It absorbs carbonic acid from 
the air and readily forms salts. The platinochloride forms 
golden-yellow plates which are quite soluble. 
In animals it produces an increase in the function of the 
skin and kidneys, and in large doses fatigue, stupor, and 
vomiting. 
Iso-amylamine, C5H13N = (CH3)2.CH.CH2.CH2.NH2, 
has been obtained by Limpricht in the distillation of horn 
with potash ; it also occurs in the putrefaction of yeast 
(Muller, Hesse, 1857); and in cod-liver oil (Gautier 
and Mourgues, 1888), where it constitutes nearly one- 
third of the bases present. 
It is a colorless, strongly alkaline liquid, possessing an 
odor which is not disagreeable. At the ordinary pressure 
it boils at 97°-98°. 
The hydrochloride forms deliquescent crystals, which 194 
BACTERIAL POISONS. 
have a bitter, disagreeable taste. The platinochloride crys- 
tallizes in goklen-yellow slender plates, which are very 
soluble in boiling water. The base is, according to Gau- 
tier and Mourgues, identical with that obtained by treat- 
ing iso-amylcarbimide with potash. 
It is a very active poison, producing rigor, convulsions, 
and death. Four milligrammes produces death in a green- 
finch in three minutes. 
Caproylamine (Hexylamine), C6H15N, has been 
found to occur by Hesse (1857) in the putrefaction of 
yeast. Hager isolated from some putrid material what he 
thought to be a mixture of amylamine and caproylamine, 
and named it septicine. 
Hexylamine was found, in small quantity, in eod-liver 
oil by Gautier and Mourgues, and according to these 
authors it resembles amylamine in its action, but is less 
toxic. 
Tetanotoxine, C5HnN, (?) was obtained by Brieger 
(1886) as one of the products of the growth of the tetanus 
microbe on beef-broth or on brain-broth. It lias also been 
obtained by Kitasato and Weye (1890) from pure cultures 
of the tetanus bacillus, kept eight days at 36°. For its 
isolation see Tetanine, and Ber. 19, 3120. It is tetanizing 
in its action, produces first tremor, then paralysis and vio- 
lent convulsions. It forms an easily soluble gold double 
salt which melts at 130°. The platinochloride is difficultly 
soluble, and decomposes at 240°. The hydrochloride is 
crystalline, and is readily soluble in alcohol and in water. 
It melts at about 205°. From warm alcohol it crystallizes 
in flat, pointed plates. 
Spasmotoxine, a base of as yet unknown composition, 
produces in animals violent clonic and tonic convulsions. 
It was obtained by Brieger (1887) from cultures of the 
tetanus germ on beef-broth. chemistry of the ptomaines. 
195 
Another toxine was obtained by Brieger (1887) in cult- 
ures of the tetanus microbe which produced complete tetanus, 
salivation, and tear-secretion. In its composition it is prob- 
ably a diamine. The platinochloride forms plates which 
begin to decompose at 240°. The hydrochloride is very 
deliquescent. Gold chloride and picric acid form very 
soluble compounds. Besides these three bases he isolated 
another toxic substance, tetanine, and a base (see under 
Tetanine). 
Dihydrolutidine, C7HuH, was found in cod-liver oil 
by Gautier and Mourgues (1888). It is the first known 
hydrolutidine. It is a colorless, somewhat oily, very alka- 
line and caustic liquid, the odor of which is sharp, but 
somewhat agreeable when dilute. It absorbs carbonic acid 
from the air, darkens and thickens ; is feebly soluble in 
water, and boils at 199° at 760 mm. pressure. The salts 
are bitter to the taste. 
The hydrochloride crystallizes in a confused mass of 
needles or in plates. The nitrate reduces silver nitrate—a 
property of all hydropvridine bases (Hofmann). The 
sulphate forms fine stellate deliquescent needles. 
The platinochloride is readily precipitated from concen- 
trated solutions as a canary-yellow precipitate. From 
warm solutions it crystallizes in lozenge-shaped plates which 
are often imbricated. On boiling with water it loses hydro- 
chloric acid and forms (C7HuNCl)2PtCl2, which possesses a 
lighter color, is more soluble than the normal salt, and crys- 
tallizes confusedly. 
The aurochloride crystallizes in needles which form fan 
or lozenge-shaped masses. It is scarcely altered even in hot 
water. 
The Iodomethylate, C7HUN.CH3I, is obtained by mix- 
ing, in the eold, the base and methyl iodide. The colorless 
compound thus obtained is soluble in water and in alcohol, 
and possesses a disagreeable, somewhat nauseating odor. 
Treated with potash it yields a colorless, aromatic, very 
alkaline oil. 
The base on oxidation with boiling potassium perman- 196 
BACTERIAL POISONS. 
ganate yields an acid, C7II7N02, and from this fact the 
discoverers conclude that the base is a dihydro-dimethyl- 
pyridine, C5H4(CH3)2NH. 
Physiological Action.—Jt is moderately poisonous. In 
small doses it diminishes the general sensibility ; in larger 
doses it produces trembling, especially of the head; pro- 
found depression alternating with periods of extreme ex- 
citement ; paralysis of the posterior limbs, and death. 
A Base, C8HuN, isomeric, but not identical, with alde- 
hyde-collidine, was obtained by Nencki as early as 1876, 
by allowing a mixture of 200 grammes of pancreas and 600 
grammes of gelatin in ten litres of water to putrefy for five 
days at 40°. The method used by Nencki for its isola- 
tion is as follows : The fluid mass was distilled with sul- 
phuric acid, to drive off the volatile acids, then rendered 
alkaline with barium hydrate, and again distilled. The 
distillate was received in dilute hydrochloric acid, and on 
evaporation gave a crystalline residue of ammonium chlo- 
ride, and of a salt which formed in long rhombic plates. 
The latter were separated from the ammonium salt by abso- 
lute alcohol. The free base was obtained from the salt by 
treating it with sodium hydrate, and extracting the solution 
with ether. 
This compound, as already stated, is isomeric with colli- 
dine, and also with O. de Coninck’s base, with which it is 
possibly identical. The latter, however, will be described 
separately. 
The free base is oily in character, and possesses a peculiar, 
not unpleasant odor. It readily absorbs carbonic acid gas 
from the air, forming after a time a lamellar, crystalline 
mass of the carbonate. The salt of this base on heating 
gives off an oil which burns with a smoky flame, and pos- 
sesses an odor similar to that of xylol or cumol. Nencki 
was therefore at first of the opinion that the ptomaine was 
an aromatic base, probably an isophcnyl-ethvlamine of the 
following composition: C6II5— Cll\^jj3. He supposed CHEMISTRY OF THE PTOMAINES. 
197 
that it was formed from the putrefaction of tyi’osin, accord- 
ing to the following equation : 
CsHuN03= CsH„N + COj + 0. 
We know that tyrosin does split up, on being heated to 
270°, into carbonic acid and oxyphenyl-ethylamine, thus : 
C.H<OT,.OH.NEr,OOOH = CtH/°= CHiNH! + 00i 
In 1883 Erlenmeyer and Lepp observed that phenyl- 
a-amido-propionic acid (phenyl-alanine), on dry distillation, 
decomposed with the formation, among other products, of a 
base having the composition C8HUN. This base was found 
to be identical with phenvl-ethylamine, C6H5.CH2.OH2.NH2, 
and in its properties and composition it resembles Nencki’s 
base. Recently (1889), Nencki has taken up a similar 
view in regard to the nature of this base, and now regards 
it as possessing the formula just given—that it is plienyl- 
ethylamine. He regards phenyl-amido-propionic acid—one 
of the three aromatic nuclei contained in the albumin mole- 
cule—as the source of this base. From the fact that phenyl- 
a-amido-propionic acid is a well-known putrefactive product, 
it would seem that Nencki’s base may arise either from the 
putrefactive decomposition of that acid, or from the splitting 
up of the acid as a consequence of the method employed in 
isolating the base. The latter would seem to be the most 
probable explanation of the genesis of this base, inasmuch 
as Brieger, by using his method for the isolation of pto- 
maines, has not been able to obtain it from putrid gelatin. 
The Platinociiloride, (C8H}1N.HCl)2PtCl4 (Pt = 
29.89 per cent.), is readily soluble in hot, and but slightly 
soluble in cold water, and can be, therefore, recrystallized 
from water. It forms beautiful flat needles. On dry 
heating it gives off* an oil which possesses an odor 
resembling very much that of xylol or cumol, and burns 
with a smoky flame. This distinguishes Nencki’s base 
from collidine, since the platinochloride of the latter does 
not show this behavior. 
encki also obtained from putrid gelatin, under certain 198 
BACTERIAL POISONS. 
ill-defined conditions, especially when no glyeoeoll was 
present, a basic product which gave, with sulphuric acid, 
large lamellar crystals. The free base forms a thick color- 
less syrup, possessing a nauseous, bitter taste. It did not 
become crystalline even after standing some time. Unlike 
the base C8HuH, it is not volatile, and is, therefore, obtained 
on evaporation of the acidulated solution after previous 
removal of the volatile bases by distillation with baryta. 
A Base, C8HuN, isomer of collidine and of the preceding 
base, with which it is possibly identical, was obtained by 0. 
de Coninck (1888) in the later stages of putrefaction of 
sea-polyps (poulpes inarins). It forms a yellowish, rather 
mobile liquid, possessing a strong benumbing (yireu&e) odor, 
and is but slightly soluble in water. It is soluble in methyl 
and ethyl alcohol, ether and acetone. Its density is 0.9865. 
When dried over potash it boils at 202° without undergoing 
decomposition. On exposure to the air it becomes brown, 
hydrates rapidly, and the boiling-point is then lowered. It 
has not been noticed to absorb carbonic acid from the air. 
It resembles some of the bases obtained from Dippel’s oil. 
The salts are in general less stable than those of the pyri- 
dine bases, and in this respect it approaches the dihydro- 
pyridine bases. 
The Hydrochloride, C8HuN.HC1, forms white or 
slightly yellowish radiate masses which are deliquescent 
and very soluble in water. The hydrobromide, ( 8HnN. 
HBr, resembles it, but is less deliquescent and a trifle less 
soluble in cold water. 
The Platinochloride, (C8H11N’.IICl)2PtCl4, is a dark 
orange-colored powder, which is insoluble, or almost so, in 
cold water, and is a rather stable compound. Boiling 
water and water at 80° decompose it into hydrochloric 
acid and (C8HnNCl)2PtCl2, which is a light-brown powder, 
insoluble in cold, scarcely so in hot water. 
The Aurochloride, C8HuN.HC1.AuC13, forms a light 
yellow precipitate. It is quite stable in cold, but very un- 
stable in hot or even warm water. It cannot be modified 
by withdrawal of hydrochloric acid. CHEMISTRY OF THE PTOMAINES. 
199 
It forms two compounds with mercuric chloride. (C8HU 
N.HCl)2HgCl2 crystallizes in small white needles, which 
are slightly soluble in water and in dilute alcohol, insoluble 
in absolute alcohol, and on exposure to moist air undergo 
change. The second compound, 2(C8HnN.HCl).3HgCl2, is 
obtained by adding an excess of concentrated mercuric 
chloride to a concentrated solution of the hydrochloride. 
It forms slightly yellow, somewhat longer needles which 
are insoluble in the principal solvents, and are likewise 
changed by atmospheric humidity. 
The Iodomethylate, C8HUN.CH3I, is formed by 
mixing solutions of the base and methyl iodide in absolute 
ether. It is deposited as a network of hue white needles, 
which are but slowly altered in the air, and are soluble in 
absolute alcohol. This solution on the addition of a little 
potash assumes a dark-red color, which is heightened by 
the addition of a little hydrochloric or acetic acid, and 
destroyed by ammonia without any resultant fluorescence. 
Warmed with excess of moist solid potash it becomes 
garnet-red in color and gives off an odor resembling that 
of the dihydropyridines. It thus behaves the same as the 
pyridine iodomethy kites. 
On oxidation with potassium permanganate it yields an 
acid which melts at 229°-230°, and begins to sublime at 
150°. It presents all the characteristics of nicotinic acid, 
c6h5no2, which is formed as the result of oxidation of 
nicotine. With hydrochloric acid it forms the compound, 
C6H5N02.HC1. With copper acetate it forms a salt; this, 
distilled with lime, yields a substance which on boiling 
with platinum chloride and water forms the compound 
(C5H5NCl)2.PtCl2. This same substance forms an iodo- 
methylate, which in alcoholic solution gives, on addition of 
potash, the characteristic reaction of pyridine bases. 
The base, C8HUN, therefore yields pyridine and nico- 
tinic acid. 
A Base, ChH13N, was obtained by Gautier and Etard 
(1881) from the chloroformic extracts (see method, page 164) 
from putrefying mackerel, as well as from the decomposing 200 
BACTERIAL POISONS. 
flesh of the horse and ox. It is regarded by these authors 
as a constant and definite product of the bacterial fermenta- 
tion of albuminoid substances, but this view is hardly justi- 
fiable, inasmuch as the base has not been found by other 
investigators. It is accompanied by the base C17II38X4 
(page 228). Nencki (1882) asserted the identity of this 
base with the one which lie had isolated in 1876, and to 
which he had ascribed the formula C8IInN. On the other 
hand Gautier and Etard consider their base to be iden- 
tical with the hydrocollidine obtained by Cahours and 
Etard by the action of selenium on nicotine. 
The free base is an alkaline, almost colorless, oily liquid, 
possessing a penetrating odor resembling that of syringa. 
It is volatile without decomposition, and boils at about 205°, 
while hydrocollidine boils at 210°. Its density at zero is 
1.0296. When exposed to the air it oxidizes slowly, be- 
comes brown and viscous, and at the same time absorbs 
carbonic acid. It differs from a collidine in possessing a 
strong reducing action, since both the gold and platinum 
double salts become reduced on heating, and even in the 
cold. 
The Hydrochloride, C8H13N.HC1, is very soluble in 
water and in alcohol, and usually forms fine needles re- 
sembling snow crystals. It is neutral in reaction and pos- 
sesses a bitter taste. In the presence of an excess of acid 
it reddens and resinifies. 
The Platinochloride, (08H13N.HCl)2PtCl4 (Pt = 
29.7 per cent.), is of a light yellow, flesh-color, crystalline, 
and but slightly soluble. It dissolves on warming, and re- 
crystallizes in bent needles. 
The Aurociiloride is rather soluble, and becomes 
slowly reduced in the cold ; rapidly on warming. 
Physiological Action.—This isomer of hydrocollidine 
is strongly poisonous. Even so small a dose as 0.0017 
gramme of the hydrochloride produced, when injected under 
the skin of a bird, marked unsteadiness of gait, followed by 
paralysis of the extremities, and finally death. The pupils 
are normal and the heart stops in diastole. Larger doses 
(0.007 gramme) cause at first vomiting and staggering, CHEMISTRY of the ptomaines. 
201 
which soon give way to a condition of exaltation. Toward 
the end tetanic convulsions set in, followed by almost com- 
plete paralysis. 
A Base, C9H13N, isomeric with parvoline, lias been ex- 
tracted by Gautier and Etard (1881) from decomposing 
mackerel and horseflesh. The method employed by these 
chemists for its isolation is given on page 164. The iden- 
tity of this base with the synthetic parvoline, obtained by 
Waage by heating ammonia with propionic aldehyde in a 
sealed tube at 200°, cannot be considered to be definitely 
settled, although an apparent identity exists in regard to 
their boiling-points. Thus, the synthetic parvoline boils at 
193°-196°, while Gautier and Etard assign to their 
base a boiling-point a little below 200°. Further investi- 
gation is necessary to decide upon the question of the iden- 
tity of this base with parvoline, or of the ptomaine C8H13N 
with hydrocollidine. 
The free base is an oily, amber-colored liquid, possessing 
the odor of hawthorn blossoms. It is slightly soluble in 
water; very soluble in alcohol, in ether, and in chloroform. 
Its boiling-point, as stated above, is a trifle below 200°. 
Like the bases C8H13N and C10H15N it becomes brown and 
soon resinifies on exposure to air. 
The Peatinocheoride, (C9H13N.HCl)2PtCl4 (Pt = 
28.65 per cent.), is slightly soluble, crystalline, and flesh 
colored; exposed to the air it soon becomes pink. 
The Aurocheoride is quite soluble. 
A Base, C10H15N, was isolated by Guareschi and 
Mosso (1883) from ox-blood fibrin which had been allowed 
to putrefy for five months. In 1887 it was re-obtained from 
putrid fibrin by Guareschi, who this time ascribed to it the 
formula C10H13N. In 1886 Oechsner de Coninck found 
it among the basic products formed in the putrefaction of 
the jelly-fish (poulpes marins, Hugounenq, page 21). The 
method used for its extraction was that of Gautier and 
Etard (see page 164). It forms a brownish oil of strong 
alkaline reaction, which soon resinifies. It possesses an 202 
BACTERIAL POISONS. 
unpleasant, weak pyridine or coniine odor, and is but 
slightly soluble in water; soluble in ether and in chlo- 
roform. 
In regard to the constitution of this ptomaine we know 
nothing, but from its physical characters it would seem to 
possess a pyridine nucleus. It is isomeric with corindine, 
a homologue of parvoline and collidine, which has been 
obtained from coal-tar. 
For the behavior of the hydrochloride to alkaloidal re- 
agents, see Table I. 
The Hydrochloride, C10H15N.HC1, crystallizes in 
colorless cholesterine-like plates which are somewhat deli- 
quescent. 
The Platinochloride, (C10H16N.HCl)2PtCl4 (Pt = 
27.52 per cent.), forms a light flesh-colored, crystalline pre- 
cipitate, and is insoluble in water, alcohol, and ether. It 
does not resinify, and is stable at 100°. 
Physiological Action.—This ptomaine resembles curara, 
although it is by no means as strong. 0.012 gramme of 
the free base produced in a frog dilatation of the pupil and 
slowing of the respiration. The nostrils were motionless, 
and within five hours complete paralysis of the muscles 
took place. The reflex excitability gradually diminished 
until it finally disappeared. An orange-blossom odor was 
observed about the frogs which were poisoned by this 
ptomaine. The same amount of ptomaine injected into a 
greenfinch produced vomiting, and a condition of weakness 
and decreased sensibility, followed soon, however, by re- 
covery. A rat was not affected by 0.020 gramme of the 
free base. The hydrochloride acts much more energetically. 
A Base, C10H15N, was isolated by 0. de Coninck, in 
1880 (Hugounenq, page 21, C. liendus, 1888), from 
sea-polyps in an advanced stage of putrefaction, together 
with the base C8HUN. The method employed for its ex- 
traction was that of Gautier and Etard (see page 104). 
It forms a slightly yellow, viscous liquid, and possesses a 
pleasant odor resembling that of blooming broom. Its 
density is about 1.18. It boils at about 230° (uncorrected), CHEMISTRY OF THE PTOMAINES. 
203 
with initial decomposition. In water it is but slightly 
soluble, readily so in ether, alcohol, acetone, and ligro'in. 
It is rapidly oxidized by the air, becomes brown, and 
resinifies but does not absorb carbonic acid. 
The Hydrochloride, C10H15N.HC1, forms line yel- 
lowish, very deliquescent needles, which in the presence of 
a trace of air are at once colored red; if more air is present 
the red changes to a brown, and in the open air a resin is 
formed the same as from the free base. It is very easily 
soluble. 
The Hydrobromide, C10H15N.HBr, crystallizes in a 
network of line deliquescent needles, which become likewise 
red on exposure to air. It is very soluble in water; less 
so in strong alcohol, and almost insoluble in ether. 
The Platinochloride, (C10H15N.HCl)2PtCl4, forms a 
dark-red powder, which is insoluble in cold water; very 
soluble in warm water. It can be kept in dry air ; in moist 
air it loses hydrochloric acid and becomes partially oxidized. 
Boiling water decomposes it. (C10H15N.Cl)2PtCl2 forms 
clear-brown plates, which are stable in moist air, and melt 
at 206°. It is insoluble in cold water, soluble in boiling 
water, but decomposes. In recrystallizing, warm previously 
boiled water should be used. 
The Aurochloride, C10H15N.HC1.AuC13, occurs as a 
light-yellow precipitate; insoluble in cold water, soluble in 
warm water. It is decomposed by boiling water ; is stable 
when kept in a moist atmosphere. 
The Iodomethylate, C10l I15Nt.CH3I, in warm alcoholic 
solution yields, on the addition of strong potash, a bright- 
red color, which soon becomes brown, and in about an 
hour the solution shows a greenish-blue fluorescence. This 
rapidity of change is due to the extreme oxidizability of the 
ptomaine. 
O. de Coninck considers tin’s base, as well as C8HUN, 
as belonging to the pyridine and not to the hydropyridine 
series. 
A Base, C10H17N, was described by Griffiths (1890) 
as derived from cultures on peptone-agar of the bacterium 204 
BACTERIAL POISONS. 
allii, a germ obtained from putrid onions. The base 
(hydrochloride?) forms colorless, prismatic, microscopic, 
very deliquescent needles, which are soluble in warm water, 
alcohol, ether, and chloroform. It gives off a hawthorn-like 
odor, especially when warmed. With phosphomolybdic acid 
it yields a white; with iodine in potassium iodide and with 
tannic acid a chestnut-colored precipitate. Nessler’s solu- 
tion produces a yellow chestnut-colored precipitate. Picric 
acid throws down a yellow slightly soluble deposit. The 
platinochloride, (C10H17N.HCl)2PtCl4, is yellow, crystalline, 
and difficultly soluble in cold water and in alcohol; soluble 
in warm water. Gold chloride produces a thick yellow 
precipitate soluble in water. Dilute sulphuric acid pro- 
duces a violet-red color. The base is apparently a hydro- 
coridine. 
A Base, C32H31N, was obtained by Del£zinier (1889) 
and is said to be the alkaloid, isolated in 1879 by Brouar- 
del, which in its chemical and physiological properties was 
described as similar to veratrine. It forms an almost color- 
less oily fluid, which possesses a hawthorn-like odor. It 
is very readily oxidizable and yields the veratrine-like re- 
actions only in the presence of air. It is soluble in alcohol, 
ether, toluene, and benzene; and forms well-defined salts 
which are very deliquescent. It appears to be an amine, 
and in its composition differs from cevadine by 9H20. 
Nothing is stated in regard to its source or method of 
preparation. The analytical results given—C = 89.41, 
11 = 7.3, N = 3.03 — correspond more to the formula 
C34H33N. 
Eth yeidenedi A mine (?), C2II8N2.—This base was con- 
sidered at first by Brieger to be identical with ethylene- 
diamine, but subsequent comparison showed this to be an 
error. Tims, the former is poisonous and does not form 
a gold salt, while the latter is not poisonous and does form 
a rather difficultly soluble gold salt. Again, ethylene- 
diamine forms a platinochloride which is almost insoluble 
in hot water, whereas the platinum double salt of the pto- chemistry of the ptomaines. 
205 
ma'ine is much more easily soluble. Brieger is, therefore, 
inclined to think that it is identical with ethylidenediamine, 
CH3.CH(NH2)2, rather than with ethylenediamine, which 
has the structure, CH2.NH2.CH2.VH2. This ptomaine 
was obtained by Brieger, in 1885 (I., 44), from decom- 
posing haddock (see Gadinine). 
The free base can be obtained, without decomposition, on 
distilling the hydrochloride with sodium hydrate. 
The Hydrochloride, C2H8N2.2HC1, crystallizes in 
long glistening needles which are readily soluble in water, 
insoluble in absolute alcohol. It gives no combination with 
gold chloride. For its behavior to alkaloidal reagents see 
Table I. 
The Platinochloride, C2H8N2.2HCl.PtCl4 (Pt = 
41.49 per cent.), forms small yellow plates which are 
moderately difficultly soluble in water. It can be readily 
recrystallized from hot water. 
Physiological Action.—Frogs seem to be less suscepti- 
ble to the action of this poison than mice or guinea-pigs. 
In the latter, it produces a short time after injection an 
abundant periodic flow of secretion from the nose, mouth, 
and eyes. The pupils dilate and the eyeballs project. 
Violent dyspnoea then comes on and predominates until the 
death of the animal, which does not take place for twenty- 
four hours or more. The heart is stopped in diastole. 
Trimethylenedi amine (?), C3H10N2 (?), is a toxic base 
isolated by Brieger (1887) from cultures of the comma 
bacillus on beef-broth. It may be stated here that from 
the same source, cholera cultures, Kunz (1888) obtained a 
base which he considered to be identical with spermine or 
ethyleneimine (see next chapter). It is present, however, in 
exceedingly minute quantity, and occurs in the mercuric 
chloride precipitate, from which it is obtained by the fol- 
lowing method : The precipitate is decomposed by hydrogen 
sulphide, the filtrate evaporated to dryness, and the residue 
taken up with absolute alcohol and precipitated by an 
alcoholic solution of sodium picrate. The precipitate 206 
BACTERIAL POISONS. 
thus obtained consists of the picrates of eadaverine, crea- 
tinine, and of this new base. It is boiled with absolute 
alcohol to remove the insoluble eadaverine picrate; the 
filtrate is evaporated to expel the alcohol, and the bases 
then converted into the platinum double salts, whereby the 
easily soluble creatinine platinocldoride can be separated 
from the corresponding less soluble compound of the new 
base. 
Owing to the small quantity of this substance present, a 
complete study of its properties has not as yet been made. 
It gives difficultly soluble precipitates with gold chloride 
and with platinum chloride ; the compound with the latter 
crystallizes in long needles. With picric acid it gives a 
precipitate consisting of felted needles, which resemble 
creatinine picrate; they melt at 198°. Phosphomolybdic 
acid yields a precipitate crystallizing in plates, while potas- 
sium-bismuth iodide gives dark-colored tine needles. From 
its physiological action it seems to be identical with the 
basic substance isolated from choleraic bodies by different 
observers. It causes violent convulsions and muscle 
tremor. 
Besides trimethylenediamine another toxinewas obtained 
by Brieger from cholera cultures, but in quantity insuffi- 
cient for analysis. It was obtained from the mercuric 
chloride filtrate after elimination of methylamine, trimethyl- 
amine, and traces of choline and creatinine, as an insoluble 
platinum double salt. Subcutaneous injection of this base 
into mice produced a paralysis-like lethargic condition, 
slowing of respiration and heart’s action, lowering of 
temperature, and finally, death in twelve to twenty-four 
hours. In some cases bloody stools were passed. 
Putrescine, C4H12N2, is a diamine which almost in- 
variably occurs together with eadaverine, with which it is 
apparently closely related. This base was also discovered 
by Brieger in 1885 (II., 42), who has obtained it from 
putrefying human internal organs (for four months at a 
low temperature without access of much oxygen); and 
from the same material, decomposing at the ordinary tern- CHEMISTRY OF THE PTOMAINES. 
207 
perature of the room, for from three days to three weeks. 
It has also been obtained from herring, twelve days in 
spring; from pike, six days in summer; from haddock, 
two months (Bocklisch). Also from putrid mussel, six- 
teen days (Brieger) ; and from human as well as horse 
flesh. Brieger has obtained it from cultures of the bac- 
teria of human feces on gelatin, and in small quantity in 
rather old cultures of the comma bacillus on beef-broth ; in 
larger quantity in cultures of the same germ on blood- 
serum. 
Udranszky and Baumann in 1888 demonstrated the 
existence of putrescine and cadaverine in the urine of cyst- 
inuria, the former constituting about one-third of the 
total amount of the two bases present. In the feces of the 
same patient, on the contrary, putrescine constituted by far 
the greater quantity, while cadaverine formed but 10 to 15 
per cent. Normal feces, as well as the feces of various 
diseases with the possible exception of cholera stools, are 
free from diamines. It would seem, therefore, that these 
bases occur in cystinuria as the result of putrefactive 
changes going on in the intestines; becoming partly ab- 
sorbed they appear in the urine. In two cases of cystin- 
uria, reported by Brieger and Stadtiiagen, cadaverine 
was found almost solely present in the urine. 
According to Mester the diamines are proportionate to 
the amount of cystin excreted, and therefore constitute a 
fixed symptom, the cause of which is the same as that of 
the cystinuria. 
Although putrescine is recognizable on about the fourth 
day of the putrefaction, yet it does not occur in appreciable 
quantity until about the eleventh day. The amount that 
is formed increases as the putrefaction goes on, so that a 
considerable quantity may be obtained after two or three 
weeks. A very good source for the preparation of putrescine, 
cadaverine, and neuridine is gelatin which has been allowed 
to decompose in contact with water for some weeks. 
Neuridine is, apparently, formed first, but is soon replaced 
by the former two bases. In the process of extraction it 
is first obtained in the alcoholic mercuric chloride precipi- 208 
BACTERIAL POISONS. 
tate. For its separation from cadaverine and other accom- 
panying bases, see page 220. 
From the urine of cystinuria it is best obtained by pre- 
cipitation with benzoyl chloride (Baumann’s method). 
For this purpose about 1500 c.c. of urine are treated 
with 200 c.c. of sodium hydrate solution (10 per cent.), 
then 20 to 25 c.c. of benzoyl chloride is added, and the 
whole shaken till the odor of the latter disappears. The 
yellowish-white precipitate which forms may consist of in- 
soluble phosphates, carbohydrates, polyatomic alcohols, and 
diamines. The cystin compound is precipitated only in 
concentrated solutions. The precipitate contains from a 
half to two-thirds of the diamines present; it is filtered 
off*, digested with warm alcohol, and the. solution filtered. 
The alcoholic filtrate is concentrated and then poured into 
about thirty times its volume of cold water. The diamine 
compounds then crystallize out. To separate the two dia- 
mines they are redissolved in just sufficient warm alcohol 
to effect solution, and this is then poured into about twenty 
times this volume of ether. The putrescine benzoyl com- 
pound is thus thrown out of solution. The filtrate from 
this on concentration yields the cadaverine compound. To 
isolate that portion of the diamines which remained in the 
original filtrate with benzoyl cystin, it is acidulated with sul- 
phuric acid and extracted with ether. The residue obtained 
on evaporating the ethereal solution is first neutralized with 
a 12 per cent, sodium hydrate solution, then mixed with 
three to four times its volume of the same solution. The 
precipitate which forms consists of the sodium compounds 
of benzoyl cystin and the diamines. It is washed with 
sodium hydrate, and the two compounds separated by their 
different solubilities in water—the cystin compound is 
readily soluble, that of the diamines insoluble. To purify 
the benzoyl diamines they are dissolved in warm alcohol 
and precipitated with excess of water. 
Putrescine (from putresco, to rot, to putrefy) is a water- 
clear, rather thin liquid which fumes in the air and has a 
a peculiar semen-like odor, almost undistinguishable from 
that of cadaverine, and reminding one somewhat of the CHEMISTRY OF THE PTOMAINES. 
209 
pyridine bases. It absorbs carbonic acid energetically from 
the air, without losing thereby the repulsive odor. The 
boiling-point of the free base, as ordinarily obtained, is 
about 135°. It is not decomposed by distillation with 
potassium hydrate, and is rather difficultly volatile with 
steam. With acids it forms beautiful crystalline salts. 
Putrescine unites with water, like ethylenediamine, to form 
a hydrate, and this water can only.be removed by distilla- 
tion with metallic sodium. The perfectly anhydrous base 
boils at 156°-157°, and then solidifies to plates (Brieger), 
which melt at 24° (Udranszky and Baumann). The 
synthetic base boils at 158°-1()0°, and melts at 23°- 
24° (Ladenburg). Like cadaverine it is difficultly solu- 
ble in ether. 
The constitution of putrescine has been determined by 
Udranszky and Baumann (1888). They showed that 
the dibenzoyl compound of putrescine was identical with 
that of the synthetic tetramethylenediamine and of the 
base which they found in the urine of cystinuria. 
Putrescine, therefore, is tetramethylenediamine, a homo- 
logue of cadaverine, and its rational formula is : 
nh2.ch2.ch2.ch2 ch2.nh2. 
The same authors (Zeitschr. f. Physiol. Chern. 13, 591) 
point out that diamines may possibly occur in putrefaction 
as the result of oxidation of monamines. Thus, putrescine 
might arise from methylamine according to the equation : 
CH3.CH2.NII2 CII2—CII2—nh2 
+ 0=| + h2o. 
ch3.ch2.nh2 cii2—cri2—nh2 
In a similar manner cadaverine might form from ethyl 
and propylamine. 
Putrescine can be prepared synthetically, according to 
Ladenburg’s method, by converting ethylene bromide into 
the cyanide and then reducing tin's by means of sodium 
in absolute alcohol. 
On heating the concentrated aqueous solution of the 
hydrochloride witli potassium nitrite there is produced an 210 
BACTERIAL POISONS. 
oil, soluble in water, from which it can be extracted with 
ether. This oil, on treatment with phenol and sulphuric 
acid, gives Liebermann’s nitroso-reaction, which would 
seem to show that putrescine is not a primary diamine 
(butylenediamine), but is rather a secondary diamine 
(Brieger, II., 42). As a primary diamine it should take 
up, on repeated treatment with methyl iodide, six methyl 
radicals; whereas, if it. is a secondary diamine, only four 
methyl radicals can enter the molecule. Thus, to illustrate, 
methylamine, CH3.NH2 (a primary amine), combines with 
three molecules of methyl iodide to form (CH3)4N.UI. 
Similarly, dimethylamine (CII3)2.NH, requires only two 
molecules to form (CH3)4N.HI. In the case of diamines, 
double this number of methyl groups is required to effect 
complete saturation. As a matter of fact, Brieger (III, 
101), on treating putrescine with methyl iodide, has suc- 
ceeded in introducing four, and only four, methyl radicals. 
From this, however, it does not follow that putrescine is 
not a primary amine, since cadaverine, an unquestioned 
primary diamine, yields a substitution compound contain- 
ing only two methyl groups (see p. 215). 
The tetra-methyl substitution-product of putrescine, 
C4H8(CH3)4N2, can be distilled without decomposition. The 
free base crystallizes in long prisms. The hydrochloride 
forms small needles which are easily soluble; with phos- 
photungstic acid it gives a white crystalline precipitate, 
with phosphomolybdic acid a yellow crystalline precipitate, 
with picric acid needles. Potassium-bismuth iodide gives 
a brownish-red amorphous deposit, while the potassium 
mercuric iodide forms prisms. Gold chloride yields diffi- 
cultly, and platinum chloride easily soluble octahedra; 
aqueous mercuric chloride forms needles. 
The aurochloride has the formula C81I22N2.2AuC14. 
This tetra-methyl derivative of putrescine is enormously 
poisonous as compared with putrescine. The symptoms 
are the same as those produced by muscarine or ncurine. 
They are: abundant salivation; dyspnoea—respiration at 
first increases, then decreases; contraction of the pupils; 
paralysisffif the muscles of the limbs and trunk; increased CHEMISTRY OF THE PTOMAINES. 
211 
peristaltic action of the intestines, ejaculation of semen, 
dribbling of urine, and, finally, violent clonic convulsions. 
In the case of mice and guinea-pigs the convulsions are 
prominent immediately after the injection of the poison. 
Putrescine Hydrochloride, C4H12N2.2HG1, forms 
long colorless needles, which are very easily soluble in 
water; difficultly so in dilute alcohol; entirely insoluble in 
absolute alcohol, and can thus be separated from cadav- 
erine hydrochloride. To accomplish this separation it is, 
perhaps, better to dissolve the mixture of the hydrochlo- 
rides in hot 96 per cent, alcohol. On cooling the solution 
thus obtained, the putrescine salt crystallizes out, whereas 
that of cadaverine remains in solution. Putrescine hydro- 
chloride differs from cadaverine hydrochloride in that it is 
not hygroscopic and can be exposed for days to the air 
without suffering any change on the surface of the crystals. 
For the behavior of the free base and the hydrochloride 
to alkaloidal reagents, see Table I. Putrescine is not toxic, 
though it possesses some marked physiological properties 
(see Cadaverine, page 215). According to Scheurlen 
putrescine, like cadaverine, produces inflammation, suppu- 
ration, and necrosis. It is not poisonous to dogs (Udran- 
szky and Baumann). It is optically inactive. 
The Platinochloride, C4H12N2.2HCl.PtCl4 (Pt = 
39.16 per cent.), often appears under the microscope in the 
form of cholesterine-like plates. In the pure condition it 
appears as six-sided plates, which are superposed in layers. 
The crystals possess a splendid silvery lustre, and are rather 
difficultly soluble in cold water; less so in hot water. 
The Aurochloride, C4H12N2.2HC1.2AuC13 + 2H20, 
crystallizes likewise in plates, which are difficultly soluble 
in cold water. It can, therefore, be readily separated from 
cadaverine aurochloride, which is easily soluble in water. 
The water of crystallization can be driven off completely 
only at 110° (B rieger). According to Bocklisch, it 
loses this water on standing over sulphuric acid, or on 
heating at 100°. 
The Picrate, C4H12H2.2C6H2(N02)30H, is difficultly 
soluble, aud crystallizes from a hot aqueous solution in 212 
bacterial poisons. 
needles; from hot aqueous alcohol, on cooling, in yellow 
plates. It begins to brown at 230°, and on further heating 
becomes darker, till finally, at 250°, it decomposes with 
rapid evolution of gas (Bocklisch). 
The Carbonate is crystalline. 
The Mercury double salt is easily soluble in a large 
quantity of water, and can thus be separated from the 
cadaverine salt, which is difficultly soluble. From hot con- 
centrated aqueous solution it crystallizes in needles. 
The Dibenzoyl - putrescine, C4H8(NHCOC6H5)2, 
forms silky plates or long needles, which are more diffi- 
cultly soluble in hot alcohol than those of the cadaverine 
compound. From this solution it is reprecipitated by ad- 
dition of water or ether. Its melting-point is 175°. It 
sublimes without decomposition. 
Cadaverine, C5HuN2, is a diamine isomeric with sap- 
rine and neuridine, and, like the latter, it occurs very fre- 
quently in decomposing animal tissues. Twelve isomers of 
this composition are possible. Another isomer, gerontine (see 
next chapter) has been described by Grandis (1890). It is 
a very striking fact, that in ordinary putrefaction as choline 
disappears the diamines appear and increase in quantity 
according as the time of putrefaction is extended. It is 
also worthy of note that cadaverine appears in putrefaction 
before putrescine. It has been obtained by Brieger (1885) 
from human lungs, hearts, livers, ete. (hence the name), 
which were allowed to putrefy at the ordinary temperature 
for three days; from the same organs, and from horseflesh, 
after four months in a closed vessel at —9° to + 5° ; 
from putrid mussel after sixteen days ; from putrid egg and 
blood albumin. It seems to be a constant product of the 
growth of the comma bacillus, irrespective of the soil on 
which it is cultivated. 
Bocklisch has isolated it from perch and pike, six days 
in midsummer; from herring, twelve days in spring ; from 
haddock, two months at a low temperature ; from cultiva- 
tions of Finkler and Prior’s vibrio proteus on beef- 
broth, thirty to thirty-five days at 37° to 38° (Ber. 20, CHEMISTRY OF THE PTOMAINES. 
213 
1441). Cadaverine seems to be a constant product of the 
activity of the genus vibrio, inasmuch as it does not occur 
in cultures in which this genus is absent. Thus, it is not 
present in the excrements of healthy or typhoid patients ; in 
cultures of Emmerich’s bacillus, of Eberth’s bacillus, and 
of the pyogenic bacteria. It is said to occur in cultures of 
the bacillus of hog-cholera (v. Schweinitz). Oechsner 
de Coninck lias found it in putrid jelly-fish (Hugounenq, 
page 23). It is present with putrescine in the urine and 
feces of cystinuria (Udranszky and Baumann, 1888). 
The odor of cholera stools and the breath of cholera patients 
may be possibly due to cadaverine, although the base has 
not been demonstrated in such cases. It has also been ob- 
tained from caviar. 
Cadaverine occurs in the mercuric chloride precipitate, 
from which it is isolated according to the methods given on 
pages 206 and 221. For its isolation and separation from 
putrescine by the use of benzoyl chloride, see page 208. 
This base was at first ascribed the formula C5H16N2, but 
subsequent researches led Brieger and Bockliscii to the 
adoption of the formula C5HT4N2. In 1883, Ladenburg 
prepared, as the first step in the synthesis of piperidine, a 
base, pentamethylenediamine, possessing the same empirical 
formula as cadaverine, and later (Ber. 18, 2956) he showed 
the possibility of the identity of these two bases. This led 
to their direct comparison and the successful establishment 
of their identity. In fact, Ladenburg, as a crucial test of 
the identity, converted cadaverine into piperidine, and found 
the latter base to agree entirely in its chemical and physical 
properties with those of the natural alkaloid (Ber. 19, 2586). 
Ladenburg, however, observed one apparent difference 
between cadaverine and pentamethylenediamine, and that 
was in the composition of the mercury double salts. That 
of the former base, whether obtained from alcoholic or 
aqueous solution (Bockliscii, Ber. 20, 1441), was found 
to combine with four molecules of mercuric chloride; 
whereas the double salt of pentamethylenediamine was 
found by Ladenburg to contain only three molecules of 
mercuric chloride. Subsequently he found that he had 214 
BACTERIAL POISONS. 
prepared this salt by mixing the aqueous solutions of the 
hydrochloride of the base and of the mercuric chloride in 
the molecular ratio of 1 to 4, and on using a larger excess 
of mercuric chloride he obtained a salt containing four 
molecules of mercuric chloride (Ber. 20, 2216). The com- 
plete identity of these two bases has, therefore, been estab- 
lished. The constitutional formula of cadaverine is, there- 
fore : 
nh2—ch2—cii2—ch— ch2—ch2—nii2. 
Cadaverine can be prepared synthetically according to 
Ladexburg’s method. For this purpose trimethylene 
bromide is converted into the cyanide, and this is then 
reduced by sodium in absolute alcohol. 
Cadaverine forms a somewhat thick, water-clear, syrupy 
liquid, which possesses an exceedingly unpleasant odor, 
resembling somewhat that of coniine (piperidine) and of 
semen. When dehydrated with potassium hydrate it boils 
at 115°-120° (Brieger). It boils at 175° (Brieger, III., 
98), and fumes in the air. The base eagerly absorbs car- 
bonic acid from the air, and solidifies into a crystalline 
mass, the carbonate. It is volatile with steam, and can be 
distilled, without decomposition, even in presence of sodium 
or barium hydrate, or soda lime. Neuridine, its isomer, 
decomposes under these circumstances. When heated with 
alcoholic potash and chloroform it does not give the iso- 
nitril reaction, nor does it give the characteristic odor of oil 
of mustard on treatment with carbon disulphide and mer- 
curic chloride. The absence of these reactions at first 
induced Brieger to conclude that cadaverine and putres- 
cine were not primary amines, but Ladenburg (1885) 
showed that this conclusion was not justifiable. These two 
reactions are given by primary monamines, but in this case 
they are not given by cadaverine, a primary diamine. It 
is probable that this behavior holds true for all diamines. 
Cadaverine is, undoubtedly, identical with the so-called 
“animal coniine,” which has been isolated at various times 
from cadavers. 
Cadaverine and putreseine were at first regarded as CHEMISTRY OF THE PTOMAINES. 
215 
physiologically indifferent, but more recent investigations 
by Scheurlen, Grawitz, and others, show that both these 
bases are capable of producing strong inflammation and 
necrosis. According to Behring, in large doses it is 
poisonous to mice, rabbits, and guinea-pigs; it is not 
poisonous to dogs (Udranszky and Baumann). Cadav- 
erine is one of those substances which can set up suppura- 
tion in the absence of bacteria. In cholera Asiatica the 
necrosis of the intestinal epithelium is quite common, and it 
would seem that this pathological change, as well as 
the muscular spasms and algidity, are due to the pres- 
ence of these bases. It should be noted, however, that 
Udranszky and Baumann failed to obtain any sign of 
intestinal irritation on feeding dogs enormous doses of 
cadaverine. Besides these local effects, they prevent, even 
in small quantity, the coagulation of blood, and render it 
“ laky.” According to Grawitz, cadaverine seems to 
hinder the growth of bacteria. The cystitis observed in 
cystinuria may possibly be due to the presence of cadaverine 
and putrescine in the urine. Both bases are optically 
inactive. 
When cadaverine is treated with methyl iodide, a base 
is obtained, the hydrochloride of which gives with pla- 
tinum chloride a double salt, having the composition: 
C5H12(CH3)2U2.2HCl.PtCl4. This new base, therefore, is 
cadaverine in which two atoms of hydrogen have been 
replaced by two methyl radicals. The platinochloride of 
this derivative forms long, clear red needles, which, unlike 
those of cadaverine, do not change their shape on repeated 
recrystallization. It is moderately difficultly soluble in 
water (Brieger, II., 41). Since cadaverine is a primary 
diamine it should combine with six molecules of methyl 
iodide to form a saturated compound. This, however, lias 
not been obtained. 
The Hydrochloride, C5H14N2.2HC1, crystallizes in 
beautiful, long deliquescent needles (Brieger). According 
to Bocklisch, it forms long, colorless needles or prisms; 
crystallizes from alcohol in plates, and is not deliquescent 
except on long standing. It is soluble in water, alcohol, 216 
BACTERIAL POISONS. 
alcohol-ether; but is insoluble in absolute alcohol, ether, 
etc. It can readily be separated from putrescine hydro- 
chloride by its solubility in 96 per cent, alcohol (Bock- 
lisch). The strictly pure base, as well as the hydro- 
chloride, does not give a blue color with ferric chloride and 
potassium ferricyanide. For reactions of the hydrochloride 
and of the free base, see Table I. 
Cadaverine hydrochloride on dry distillation decomposes 
into NII3, HC1, and piperidine, C5HUN. The latter is a 
well-known poisonous alkaloid which exists in the combined 
state in black pepper. It is not known whether this change, 
whereby the non-poisonous cadaverine is converted into a 
toxic base, can take place under the influence of bacteria 
during the processes of putrefaction or not. However, it 
does not seem improbable that this simple chemical change 
should be effected through the action of living organisms; 
for Schmidt has already shown that the almost physiologi- 
cally indifferent choline, when subjected to the action of 
the bacteria of hay-infusion, decomposes into a neurine-like 
base possessing a muscarine-like action, and under certain 
conditions it yields a base which in its action resembles 
pilocarpine. 
The Sulphate likewise forms beautiful, well-formed 
needles, and in its solubility corresponds to the hydro- 
chloride. 
The Platinochloride, C5H14N2.2HCl.PtCl4 (Pt = 
38.08 per cent.), crystallizes after some time, on the addition 
of platinum chloride to a not too concentrated solution of the 
hydrochloride, in the form of long, beautiful orange-red 
needles (Bocklisch). Ordinarily it is obtained at first in 
long, dirty red needles, which on repeated recrystallization 
become clearer and assume a form similar to that of ammo- 
nium platinochloride. It forms chrome-yellow rhombic 
prisms which are short and octahedra-like. In polarized 
light they are strongly double refracting. It is very slightly 
soluble in cold water; can be recrystallized from hot water 
(Bocklisch). Its solubility in water at 12° is l'to 113- 
114. It decomposes at 235°-236°. 
The Aurochloride,C5H14N2.2IIC].2AuC13(Au=50.41 chemistry of the ptomaines. 
217 
per cent.), crystallizes partly in cubes, and partly in long 
needles which at first possess a bright lustre, but under the 
desiccator soon effloresce and become opaque. The water 
of crystallization is completely removed on standing over 
sulphuric acid. It is very easily soluble, and melts at 188° 
(Bocklisch). 
The Picrate, C5H14N2.2C6H2(N02)30H, forms yellow 
plates which are difficultly soluble in cold water. From 
hot water it crystallizes in long prisms, which melt at 221° 
with decomposition. It is insoluble in absolute alcohol 
and can be recrystallized from hot dilute alcohol. 
Cadaverine hydrochloride combines with mercuric chlo- 
ride, when the aqueous solutions of these two salts are 
mixed in the molecular ratio of 1 to 4, to form c5h14n2. 
2HC1.3IIgCl2. This salt can be recrystallized from hot 
water (Ladenburg). When an excess of mercuric chlo- 
ride is used the double salt has the composition C5H14N2. 
2IIC1.4HgCl2. This last salt melts at 216° (Ladenburg) ; 
at 214° (Bocklisch). It is difficultly soluble in cold 
water; from hot water it crystallizes in needles or plates 
(Bocklisch). 
The Neutral Oxalate, C5H14N2.H2C204-j-2H20, was 
prepared by Bocklisch by adding a little less than the cal- 
culated quantity of alcoholic oxalic acid to the cadaverine. 
The precipitate may be recrystallized from hot dilute alco- 
hol, when it is obtained in the form of needles, which melt 
at about 160° and at the same time give off gas. 
The Acid Oxalate, C5H14N2.2H2C204+H20, is made 
by bringing the neutral salt into alcoholic oxalic acid. It 
• is soluble in hot dilute alcohol, and recrystallizes from it in 
quadratic plates, sometimes in glistening needles. It melts 
at 143° with decomposition. After it has been dried 
over sulphuric acid, it loses, on being heated to 105°-110°, 
one molecule of water (Bocklisch, Ber. 20, 1441). The 
insolubility of these oxalates in absolute alcohol shows the 
fallacy of Tamba’s distinction between ptomaines and vege- 
table alkaloids. (See page 186.) 
The dibenzoyl derivative, C5H10(NHCOC6H5)2, crys- 
tallizes in long needles and plates, readily soluble in 218 
BACTERIAL POISONS. 
alcohol, difficultly so in ether, and insoluble in water; 
hence the alcoholic solution can be precipitated by addition 
of water or ether (separation from the putreseine compound, 
see p. 208). It melts at 129°-130°. It is not changed by 
boiling with dilute acids and alkalies; but boiling with 
concentrated hydrochloric or sulphuric acids for a long 
time finally breaks it up. 
Neuridine, C5H14N2, was tlie first diamine isolated from 
animal tissues (Brieger, 1883). It is one of the most 
common products of putrefaction, and as such has been 
obtained by Brieger from putrid horseflesh, beef, human 
muscle, five to six days; from haddock, five days in sum- 
mer ; from cheese, six weeks in summer; from gelatin, 
ten days at 35° ; from decomposing human internal organs, 
three to eleven days ; from cultures of the Eberth bacillus, 
with mydine. 
Bocklisch has obtained it from perch, six days in 
summer ; from barbel after three days in summer. 
It has also been obtained from fresh eggs in the prepa- 
ration of choline by heating with baryta; and also from 
fresh brain by heating with 2 per cent, hydrochloric acid 
(Brieger, I., 57-61). Ehrenberg (1887) found it in 
poisonous sausage and obtained it by growing a bacillus 
from this source on liver and meat bouillon. 
Neuridine is almost invariably accompanied by choline, 
and as the duration of putrefaction increases, the latter 
gradually decreases in amount and yields a corresponding 
increase in trimethylamine, whereas the yield of neuridine 
increases from day to day. The amount of neuridine 
formed depends upon the nature of the organ employed in 
putrefaction. The greatest yield is obtained from gelatin- 
ous tissues, such as intestines; and especially from pure 
gelatin. On the other hand, such tissues as the spleen 
and liver yield but little. 
Neuridine comes down in the mercuric chloride precipi- 
tate (sometimes it occurs in the filtrate), and can then be 
isolated from the other bases present in a number of ways. 
One method is given under Gadinine. Another convenient CHEMISTRY OF THE PTOMAINES. 
219 
method of separation is to precipitate it from alcoholic 
solution by alcoholic picric acid. The picrate thus ob- 
tained is, for the purpose of further purification, recrystal- 
lized from absolute alcohol, then decomposed by extracting 
its acid solution with ether (to remove the picric acid) and 
evaporating the aqueous solution to dryness. The residue 
is now extracted with alcohol and the alcoholic solution 
precipitated by alcoholic platinum chloride. The platino- 
chloride can now be recrystallized from hot water. 
The free base, as obtained by the treatment of the 
hydrochloride with moist freshly precipitated silver oxide, 
possesses an extremely repulsive odor, similar to that of 
human semen. On evaporation of its aqueous solution it 
yields a gelatinous-like mass, and at the same time slowly 
decomposes. It does not crystallize when evaporated in a 
vacuum, and decomposes even under these conditions. The 
same disagreeable odor is obtained when the hydrochloride 
is warmed with potassium hydrate. Brieger (I., 24) re- 
gards this decomposition-product of neuridine as au oxida- 
tion product of the original substance. 
The free base is very readily soluble in water, but is 
insoluble in ether and absolute alcohol; difficultly soluble 
iu amyl alcohol. It gives white precipitates with mercuric 
chloride, neutral and basic lead acetates. When distilled 
with fixed alkali it yields di- and tri-methylamine, thus 
probably showing some relation to neurine, hence the name 
neuridine. It does not give Hofmann’s iso-nitril reac- 
tion. but it does not follow from this, as shown under 
cadaverine, that it may not be a primary diamine. It is 
isomeric with cadaverine, saprine and gerontine. 
The Hydrochloride, C5H14N2.2HC1, crystallizes in 
long needles which are extremely soluble in water and 
in dilute alcohol, but are insoluble in absolute alcohol, 
ether, benzol, chloroform, petroleum ether, benzine, amyl 
alcohol, etc. Its insolubility in absolute alcohol may be 
used to effect a separation from choline hydrochloride. It 
can be recrystallized from slightly warm dilute alcohol. 
Although the pure salt is insoluble in the reagents just 
giveu, nevertheless, in the presence of other animal matter 220 
BACTERIAL POISONS. 
it is dissolved in greater or less quantity, and hence can be 
obtained by the Stas-Otto as well as by the Pragen- 
dorff method. The crystals resemble urea in form. On 
heating very cautiously the salt sublimes, and at the same 
time appears to undergo a partial internal decomposition, 
inasmuch as many of the groups of needles in the sublimate 
are colored red or blue. For the behavior of the hydro- 
chloride with the alkaloidal reagents, see Table I. 
Pure neuridine is not poisonous, but as long as it is 
contaminated with other putrefaction products it possesses a 
toxic action similar to that of peptotoxine. This holds 
true for the other non-poisonous bases. . 
The Platinochloride, C5H14N2.2HCl.PtCl4, crystal- 
lizes in beautiful flat needles. Recrystallized from hot 
water, it forms aggregations of small, clear, yellow needles. 
It is readily soluble in water, from which it is precipitated 
on the addition of alcohol. 
The Aurochloride, CsH14N2.2HC1.2AuC13, is rather 
difficultly soluble in cold water (Bocklisch), and crystal- 
lizes on cooling of the hot, saturated solution in bunches of 
clear, yellow, short needles. 
The Picrate, C5H14N2. 2C6II2(X02)3OII, can be recrvs- 
tallized from boiling water, in which it is very difficultly 
soluble, in the form of needles united in plumose groups. 
It is almost insoluble in cold water ; less difficultly soluble 
in alcohol. It is not fusible, but begins to brown and 
give off yellow vapors at 230°, and carbonizes completely 
at 250°/ 
Saprine, C5H14N2, was found in human livers and 
spleens after three weeks’ putrefaction (Brieger, II., 30, 
46, 58). It occurs together with cadaverine, putrescine, 
and mydaleine in the mercuric chloride precipitate. To 
separate these bases, Brieger (1885) used the following 
process : The mercury salts were decomposed with hydrogen 
sulphide, the filtrate evaporated to dryness, and the residue 
then extracted with alcohol. The putrescine hydrochloride 
is insoluble in alcohol, and is thus removed. The alcoholic 
solution was treated with platinum chloride, which precipi- CHEMISTRY OF THE PTOMAINES. 
221 
tated the greater part of the cadaverine. The mother- 
liquor, on concentration, yielded a mixture of the platino- 
chlorides of cadaverine and saprine. Each successive crop 
contained more of the saprine double salt. The two kinds 
of crystals were now separated by means of a magnifying- 
glass. The saprine platinoehloride thus obtained was finally 
purified by repeated recrystallization from water. The 
mother-liquor, after the removal of the saprine platino- 
chloride, contains the mydaleine salt, which, on account of 
its solubility in water, crystallizes only on concentration, 
or on standing under a desiccator. The mercuric chloride 
filtrate contains some mydaleine and the ptomaine, which 
yields a platinoehloride containing 28.40 per cent, platinum. 
The free base is a diamine, and was first ascribed the 
formula C5II16N2. It appears, however, to be isomeric 
with cadaverine and neuridine. The term saprine is derived 
from the Greek <?airp6c} signifying putrid. It possesses a 
weak pyridine-like odor, and can be distilled with steam or 
with potassium hydrate without undergoing decomposition. 
In its reactions it behaves the same as cadaverine, except 
that it gives an amorphous precipitate with potassium- 
bismuth iodide, whereas cadaverine gives a crystalline pre- 
cipitate. The free base gives an immediate intense blue 
color with ferric chloride and potassium ferricyanide. 
The Hydrochloride, C5H14N2.2HC1, forms flat needles 
which are not hygroscopic (distinction from cadaverine 
hydrochloride). Its reactions are the same as those of 
cadaverine hydrochloride (see Table I.). It is, however, 
tinged slightly blue by a mixture of ferric chloride and 
potassium ferricyanide, whereas the free base gives an 
intense blue. It differs from cadaverine in that it does not 
give the reddish-brown color with potassium bichromate 
aud sulphuric acid. Again, it forms no aurochloride; 
while, on the other hand, cadaverine hydrochloride yields 
an easily soluble salt, crystallizing in splendid needles. 
The Platinochloride, C5H14N2.2HCl.PtCl4, forms 
parallel, aggregated, pointed crystals, which are somewhat 
soluble in water, and are thus distinguished from cadaverine 222 
BACTERIAL POISONS. 
platinochloride, which crystallizes in rhombs, and is diffi- 
cultly soluble in water. 
Physiologically, it is indifferent. 
A Base, C7H10N2.—Until very recently the nature of the 
basic substances which are formed as products of the alco- 
holic fermentation of sugar or molasses has been but little 
understood. Kramer and Pinner, in 1869, found in crude 
fusel oil a small quantity of a volatile base which they 
apparently identified with a collidine. This observation was 
confirmed by Ordonneau and others; and still more re- 
cently (January, 1888) Morin has contributed an elaborate 
paper upon the bases formed during alcoholic fermentation. 
The portion of crude fusel oil which boils above 130.5° 
was extracted with slightly acidulated water, the acid 
aqueous solution thus obtained was made alkaline, and the 
oily bases which were thus set free were then distilled with 
vapor of water. The free bases were dried over potassium 
hydrate and then subjected to fractional distillation. Three 
fractions were thus obtained, boiling respectively at 155°- 
160°, 171°-172°, and 185°-190°.‘ Only the second frac- 
tion, which boils at 171°-172°, was studied, and was found 
to possess the formula C7H10N2. Heated with concentrated 
hydrochloric acid, it is decomposed in part with the forma- 
tion of ammonia. It combines with ethyl iodide to form a 
yellow crystalline compound, which is soluble in water 
and alcohol, insoluble in ether. The hydrochloride crys- 
tallizes in fine white needles, soluble in water and alcohol, 
and but very slightly soluble in absolute ether. The free 
base, as stated above, boils at 171°-172°, is very soluble in 
water, alcohol, ether, etc. When pure it forms a colorless, 
strongly refracting, very mobile oil, which possesses a char- 
acteristic nauseating odor, but slightly resembling that of 
the pyridine bases. Its density at 12° is 0.9826; toward 
litmus paper the base shows no decided reaction. The 
platinochloride is crystalline and is very soluble in water 
and alcohol, slightly soluble in ether. Potassio-mercuric 
iodide does not precipitate the aqueous solution of the free 
base, but in solutions of the hydrochloride it gives a yellow 223 
CHEMISTRY OF THE PTOMAINES. 
flocculent precipitate, which soon crystallizes in long bril- 
liant yellow needles. This reaction takes place readily in 
solutions of 1 to 1000, and only after some hours in solu- 
tions of 1 to 10,000; and is not given by the bases of the 
pvridic and quinolinic series. Mercuric chloride produces 
an immediate flocculent precipitate in solutions of the base 
having a concentration of 1 to 1000, but requires some 
time to appear in 1 to 10,000. Phosphotungstic acid gives 
an immediate white precipitate even in a dilution of 1 to 
10,000. Phosphomolybdic acid in solutions of the same 
strengtli yields a yellow precipitate. 
The physiological action of this base has been examined 
by R. Wurtz, who found the lethal dose for rabbits, etc., 
to be about one gramme per kilogramme of body weight. 
It produces stupor, paralysis, which at first appears in the 
rear extremities; the sensibility becomes diminished and 
the pupils are dilated and unresponsive to light; the rate 
of heart-beat is lowered, and the rectal temperature falls as 
low as 35° ; death follows a more or less prolonged coma. 
Tax ret obtained by the action of ammonia on glucose 
a number of bases, to which he applied the generic name 
of glucosines. One of these, having the formula C14H10N2 
(C = 6), corresponds in its formula and its general proper- 
ties to Morin’s base C7H10jST2 (C = 12), and, in fact, the 
two bases are considered by Tanret to be identical. 
It is interesting to note in this connection that alkaloidal 
bases have been found in petroleum by Bandrowski, and 
that similar basic substances have been detected by Weller 
in paraffin oil. 
Most of the solvents in common use, such as alcohol, 
ether, chloroform, benzole, petroleum ether, amyl alcohol, 
etc., have been shown at different times to contain basic 
pyridine compounds, though ordinarily in very minute 
quantity. On the other hand, Haitinger has found in 
some specimens of amyl alcohol as much as 0.5 per cent, 
of pyridine. 
Susotoxine, C10H26N2 (?), is a base isolated by Now in 
1890 from cultures of the hog-cholera bacillus of Salmon 224 
BACTERIAL POISON8. 
(swine-plague of Billings). It is probably identical with 
the base obtained by v. Schweinitz from the same germ, 
although the formula ascribed to it by him is C14H32N2. 
The free base lias not been obtained. The hydrochloride 
forms a light-yellow syrup which shows no tendency to 
crystallize. It is soluble in water and in absolute alcohol, 
and is somewhat hygroscopic. When heated with fixed 
alkali it gives off a strong amine odor, such as is perceived 
on evaporating the original culture-fluid, if it happens to be 
alkaline in reaction. 
The platinochloride is obtained by precipitation as a light, 
flesh-colored, granular precipitate. It is readibly soluble 
in water, from which it can be reprecipitated by addition 
of absolute alcohol. From aqueous solution, when allowed 
to evaporate slowly, it crystallizes in long, thick needles. 
The mercurochloride is thrown down from solutions of 
the hydrochloride in absolute alcohol, by alcoholic mercuric 
chloride, as a heavy, white, granular precipitate. This 
readily dissolves on the addition of a small quantity of 
water, and can be perfectly reprecipitated by addition of 
absolute alcohol. On treatment with hydrogen sulphide it 
is readily decomposed, yielding the pure hydrochloride 
The aurochloride is very soluble in water and alcohol. 
From the alcoholic solution it may be partially precipitated 
by ether as a light-yellow, oily precipitate, which is adhe- 
rent to the sides and bottom of the tube. 
Physiological Action.—The base is toxic; only in rela- 
tively large doses, as seen from the following experiment. 
About 100 milligrammes, dissolved in a little water, were 
injected subcutaneously into a young rat. The animal 
was at first quiet, apparently unwilling to move. After 
some ineffectual attempts at jumping, it settled down in a 
recumbent position, and when placed on its side was unable 
to rise. Respiration was at first retarded, later increased, 
but toward the end was again very slow. Convulsive tre- 
mors shook the body at frequent intervals. The animal 
kicked vigorously. Reflexes were present almost to the 
end. As death approached, the red eyes whitened and took 
on a glazed, opaque appearance. Death resulted in one CHEMISTRY OF THE PTOMAINES. 
225 
and a half hours. The animal was on its side, the feet 
extended. Post-mortem examination showed the heart 
arrested in diastole, lungs rather pale, stomach contracted, 
serum in thoracic cavity, subcuta pale and (edematous. 
Repeated doses of smaller quantities seem to confer a partial 
immunity to the action of the germ. 
Methyl-guanidine, C2H7N3, =NH= 3‘ 
This base has long been known as a product of the oxi- 
dation of creatine and creatinine, but had never been met 
with in animal tissues. Brieger in 1886 (III., 33) ob- 
tained it from horseflesh which was allowed to decompose 
in a closed vessel at a low temperature (—9° to +5°) for 
four months. Bockliscii (Ber. 20, 1441) isolated it from 
impure cultures on beef-broth of Finkler and Prior’s 
vibrio protens, containing ordinary putrefaction bacteria, 
for twenty to thirty days at 37°-38°. Vibrio proteus 
alone seems incapable of forming this base. The comma 
bacillus after some time (six weeks) partially decomposes 
creatinine with formation of a small quantity of methyl- 
guanidine (Brieger). The bacillus of anthrax likewise is 
capable of transforming creatine into methyl-guanidine. 
It occurs in the mercuric chloride filtrate (Brieger), 
from which it is obtained, after the removal of the mercury 
by hydrogen sulphide, by precipitation with phospho- 
molybdic acid. The precipitate is decomposed with neutral 
lead acetate, and the filtrate from this, alter removal of the 
lead by hydrogen sulphide, is concentrated and then sodium 
picrate added. The resinous picrate precipitate is purified 
by boiling with much water, and, finally, it is recrystallized 
from boiling absolute alcohol. According to Bockliscii, 
it occurs in the mercuric chloride precipitate (not in the 
filtrate), from which it is isolated, after removal of the mer- 
cury and concentration of the clear filtrate, by precipitation 
with sodium picrate. The precipitate containing cadaverine, 
methyl-guanidine, and creatinine, is boiled with absolute 
alcohol (cadaverine picrate is insoluble) and the alcoholic 
solution is then evaporated to drive ofl the alcohol and 226 
BACTERIAL POISONS. 
taken up with water. From this aqueous solution, after 
removal of picric acid, methyl-guanidine is precipitated by 
gold chloride, whereas creatinine remains in solution. 
This ptomaine is identical with the synthetic methyl- 
guanidine (methyluramine) which can be readily obtained 
by boiling a creatine solution with mercuric oxide or with 
lead dioxide and dilute sulphuric acid (Dessaignes). The 
parent substance of methyl-guanidine as it occurs in putre- 
faction is undoubtedly the creatine which exists preformed 
in the muscular tissue. If such is the case, the bacteria 
engaged in its production must be considered as possessing 
an oxidizing action, since this base is prepared synthetically 
from creatine by oxidation. That creatine does not offer 
much resistance to the action of bacteria is shown in the 
fact that Friedlander’s pneumonia coccus, which pos- 
sesses but small chemical powers, is capable of slowly but 
steadily decomposing creatine, yielding as one of the pro- 
ducts acetic acid. Strecker and Erlenmeyer, as well 
as Baumann, have shown that creatine, although a sub- 
stituted guanidine, is not poisonous, but is readily converted 
into creatinine, which is a relatively toxic substance. On 
the other hand, guanidine and methyl-guanidine are quite 
violent poisons. This is, therefore, another instance in 
which a toxic substance is formed by the action of bacteria 
from a previously non-poisonous base (see page 244). 
According to Lossen, guanidine is formed, though in 
small quantity, in the oxidation of albumin. 
The formulae of these closely related substances are here 
given for comparison : 
Creatine, NH = C/N(CH3).CH2.C02H 
/N(CH3).CH2 
Creatinine, NH — C\j^jj 
/N(CH3).CH2 
Methyl-hydantome, () = 
Methyl-guanidine, NH==C(^Tj^'^ chemistry of the ptomaines. 
227 
Guanidine, NII = \ y[H.2 
Urea, 0 = 0<£g; 
Methyl-guanidine forms a colorless, easily deliquescent 
mass possessing a strong alkaline reaction. On heating 
with potassium hydrate it decomposes, and yields ammonia 
and methylamine. It is a highly poisonous base. 
The Hydrochloride, C2H7N3.HC1, can be obtained 
from the picrate by dissolving the latter in water acidulated 
with hydrochloric acid, and extracting the solution with 
ether to remove the picric acid. The colorless aqueous 
solution now, on evaporation, yields a thin syrup which 
crystallizes in vacuum to compact prisms. These are in- 
soluble in alcohol, and give with platinum chloride a double 
salt of monoclinic needles (Haushofer) which are very 
easily soluble (1 part in about 7 parts water, Tatarinow). 
The Aurochloride, C2H7N3.HC1.AuC13 (Au = 47.71 
per cent.) forms rhombic crystals (Haushofer) which are 
easily soluble in ether, more difficultly in water or alcohol; 
readily soluble (Brieger). It readily decomposes on heat- 
ing in pure water, but may be recrystallized from water 
acidulated with hydrochloric acid. It melts at 198°. 
The Picrate, C2H7N3.C6H2(N02)30H, comes down at 
first as a resinous precipitate, which when boiled with much 
water solidifies in the form of felted needles. It is very 
difficultly soluble in water, and can be purified by repeated 
recrystallization from boiling absolute alcohol—distinction 
from cadaverine. It melts at 192°. 
The Oxalate, (C2H7N3)2.H2C204-|-2H2O, forms crystals 
which are easily soluble in water. 
Physiological Action.—Methyl-guanidine as obtained 
from putrefying flesh is identical in its physiological 
action with the synthetic base. It has already been stated 
that the non-poisonous creatine is readily converted into the 
relatively energetic poison creatinine. The latter substance 
possesses a paralyzing action differing very much from its 228 
BACTERIAL POISONS. 
decomposition-product methyl-guanidine. This base is very 
poisonous, and the symptoms are marked by dyspnoea, 
muscle tremor, and general clonic convulsions. B rieger 
lias observed the following symptoms on injection of about 
0.2 gramme of methyl-guanidine into a guinea-pig: The 
respiration at once becomes more rapid, and in a few min- 
utes abundant passage of urine and stool takes place; the 
pupils dilate rapidly to the maximum and cease to react. 
The animal is uneasy but motionless, though not exactly 
paralyzed. Respiration becomes deeper and more labored, 
the head moves from side to side, the extremities become 
gradually paralyzed; dyspnoea sets in, the animal falls on 
its side and dies (twenty minutes) amid general clonic con- 
vulsions of short duration. Fibrillary twitchings of the 
trunk muscles are observed only in the beginning. Post- 
mortem showed the heart to be stopped in diastole, the in- 
testines filled with fluid, the bladder contracted, the cortex 
of the kidney hypercemic, but the papillae of the kidneys 
surprisingly pale. 
Morrhuine, C19H27N3, was obtained by Gautier and 
Mourgues (1888) from the mother liquors of aselline on 
concentration of the platinum-containing liquid. This sub- 
stance constitutes about one-third (0.07 per cent.) of all the 
bases found in cod-liver oil, and is named from Gadus 
morrhua, the ordinary codfish. The free base is an oily, 
very thick, amber-yellow liquid, the odor of which resem- 
bles somewhat that of syringa. It floats on water and par- 
tially dissolves; is more soluble in ether and in alcohol. 
The base is very alkaline and is caustic to the tongue. It 
absorbs carbonic acid and is non-volatile. The salts of 
copper are precipitated by it, but the hydrate formed is not 
redissolved 
The hydrochloride is very deliquescent. The gold salt 
forms a yellow precipitate which readily dissolves on 
warming. The platinum salt, C19H27N3.2HCl.PtCl4 (Pt = 
27.56 per cent.), crystallizes in barbed needles, which are 
quite soluble. (Separation from aselline, p. 230). CHEMISTRY OF THE PTOMAINES. 
229 
Physiological Action.—The base possesses the property 
of exciting the appetite; it acts as a diaphoretic and above 
all as a diuretic. 0.029 gramme given subcutaneously 
to a guinea-pig produced in two and a half hours a loss 
of 13.5 grammes in the weight of the animal. The same 
effect is produced in birds. Strong doses (0.1 gramme per 
kilogramme) produce fatigue and hebetude. 
A Base, C131I20V4, was obtained as early as 1868 by 
Oser, who observed its formation during the fermentation 
of pure cane-sugar by means of yeast. The hydrochloride 
when dried in vacuo is said to form a white, very hygro- 
scopic foliaceous mass, which soon becomes brown on expo- 
sure to air. At first it imparts a burning taste, which is 
soon replaced by a very bitter sensation. 
A Base corresponding to the formula Cl7H38N4 was ob- 
tained by Gautier and Etard from the mother-liquors of 
the platinochloride of the base C8H13N. Very little is 
known, however, in regard to the general properties of this 
base, owing to the small quantity which could be isolated. 
This base and the one obtained by Oser from the yeast- 
fermentation of sugar, C131I20N4, and aselline, t3 v 
are the only ptomaines thus far isolated which are known 
to contain four atoms of nitrogen. 
The Platinochloride, C17H38N4.2HCl.PtCl4 (Pt = 
27.62 per cent.), is readily soluble, and crystallizes in 
needles which possess a light-yellow flesh color. When 
heated to 100°, it slowly decomposes, giving off a syringa- 
like odor. 
Aseltjne, C25H32N4, isolated by Gautier and Mour- 
gues (1888), together with five other bases from cod-liver 
oil. (See p. 263.) It is present only in small quantity in 
the oil. The name is derived from Asellus major, the great 
codfish. The free base is thrown down from the1 solutions 
of the hydrochloride by the addition of alkali, in amorphous 
white floccules which are almost insoluble in water. It is 
almost colorless, but on exposure to the air becomes slightly 230 
BACTERIAL I’OISONS. 
green. It is not hygroscopic, and possesses a density of about 
1.05. On heating it melts to a viscid yellowish fluid, pos- 
sessing an aromatic odor; is non-volatile. Although almost 
insoluble in water, it imparts to it an alkaline reaction and 
a bitter taste. It is soluble in ether, more so in alcohol. 
The salts are crystal lizable, but are partially dissociated 
by the action of warm water. The hydrochloride forms 
crossed or entangled needles which are quite bitter. The 
gold salt is very reducible. The platinochloride, C25H32N4. 
2HCl.PtCl4 (Pt = 24.41), is orange-yellow in color; solu- 
ble in warm waler, insoluble in cold water (separation 
from morrhuine, p. 228), and is rapidly changed by boiling 
water. The mercury salt is precipitated in the cold ; redis- 
solves on heating, and then, on cooling, recrystallizes. 
In large doses it produces fatigue, short and rapid respi- 
ration, and stupor. Three milligrammes of the hydro- 
chloride kills a greenfinch in fourteen minutes. 
Mydine, C8HuNO, is a non-poisonous base which 
was obtained by Beieger in 1886 (HI., 25) from the 
putrefaction of about two hundred pounds of human in- 
ternal organs; and also in cultures of the Eberth bacillus 
on peptonized blood-serum. It occurs in the mercuric 
chloride filtrate, and is isolated from it after the removal 
of the mercury by hydrogen sulphide, by precipitation with 
phosphomolybdic acid. The gummy precipitate which is 
produced is decomposed on the water-bath with a solution 
of neutral lead acetate, and the filtrate on evaporation yields 
a colorless hydrochloride, crystallizing in plates. It is 
purified by recrystallization of the pi crate. 
The free base is strongly alkaline, and possesses an am- 
moniacal odor. It is characterized bv its strong reducing 
properties. The name mydine is derived from fivSau, to 
putrefy. With platinum chloride it gives, after a time, an 
extremely soluble salt; with gold chloride, a precipitate of 
metallic gold. On distillation it is decomposed. 
The Hydrochloride, C81IuNO.HC1, crystallizes in 
colorless plates. It gives a blue color with ferric chloride 
and potassium ferricyanide. CHEMISTRY OF THE PTOMAINES. 
231 
The Picrate, CsIIuN0.C6H2(NO2)3OfI, is obtained in 
broad prisms, which melt at 195°. It is the only salt 
suitable for manipulations. 
In describing Nencki’s collidine (page 196) it was stated 
that tyrosin might be looked upon as the source of that 
base. It would seem, however, to be more appropriately 
the parent substance of mydine, inasmuch as it decomposes 
on being heated to 270° into carbonic acid and oxyphenyl- 
ethylamine, C8HuNO. The change that takes place can 
be represented by the equation : 
c-ii<<cE,.chnh1.co1h = + CO, 
Tyrosin. 
OxYPHEN YI.-ETH YI.AMINE. 
A Base, C5HuN02, was isolated by E. and H. Sal- 
kowski (188-3) from decomposing fibrin and meat. In its 
composition it is isomeric with betaine anhydride. It is 
extremely soluble in water, very difficultly so in alcohol, 
insoluble in ether, and possesses a semen-like odor and 
saline taste. The aqueous solution, which is not alkaline 
in reaction, yields on evaporation a stellate crystalline mass, 
which on standing over sulphuric acid becomes a white 
powder, which melts at 156°. It dissolves silver oxide, 
but not cupric hydrate, thus apparently indicating that it 
is not an amido acid. Moreover, it does not give a pre- 
cipitate or blue coloration with copper acetate, or ammo- 
niacal silver nitrate. It thus differed from the then known 
amido-valerianic acids, its isomers. Recently, however 
(1891), Gabriel and Aschan showed that <5-amido-vale- 
rianic acid agrees with this base in its reactions to copper 
and silver oxide, copper acetate, and ammoniacal silver 
nitrate. The gold salt of the synthetic base possessed the 
same composition as that of 8alkow.sk r, and melted at 
86°-87°. 
The identity of this base with -am i do - val er i an i c acid 
(homopiperidinic acid) would seem to be established, and 
as such it is regarded. Its structure, then, is represented by 
NII2.CH2.CH2.CH2.CH2.C02H. 232 
BACTERIAL POISONS. 
For its synthetic preparation see Ber. 24, 1365 (1891). 
The base does not seem to possess a toxic action. 
The Hydrochloride, C5IIuN02.HC1, forms colorless, 
stellate crystals, which are permanent in the air, and are 
extremely soluble in water, even in absolute alcohol. 
The Aurochloride, C5HuN02.HC1.AuC13 -f- II20, is 
obtained on slow evaporation, as large, well-formed, beau- 
tiful dark-yellow crystals. They are probably monoclinic, 
contain water of crystallization, and melt at below 100°. 
T1 ie Platinochloride gave on analysis results cor- 
responding to the formula (C7H15N02.HCl)2PtCl4. This 
may possibly be due to the presence of some higher homo- 
logues of the base C5HuN02. It forms fine orange-yellow 
crystals, which are very difficultly soluble in alcohol, easily 
so in hot water, from which, on cooling, it crystallizes in 
beautiful plates. 
Choline Group.—The following four bases are closely 
related, and, indeed, starting from choline, the oldest and 
best-known individual, the remaining bases can be readily 
prepared from it. Moreover, they can all be prepared 
synthetically according to methods that will be subsequently 
indicated. As choline is the most prominent member, we 
have thought best to class these substances together as con- 
stituting the choline group. It is very probable that my- 
datoxine and mytilotoxine, when their constitution becomes 
known, will be found to be homologues of certain members 
of this group. 
Neurine, C5H13NO = C2H3.N(CH3)3.OH. — This sub- 
stance was obtained and named thus by Liebreich (1865), 
who prepared it by boiling protagon for twenty-four hours 
with concentrated baryta. Previous to its discovery as a 
decomposition-product of protagon from the brain it was 
prepared synthetically bv IIofmann (1858) by treating 
trimethylamine and ethylene bromide with potassium hy- 
drate or silver oxide. Ha eye R (1866), by boiling an alco- 
holic. extract of the brain with baryta water, obtained on 
separation by three different methods, a base, or rather a CHEMISTRY OF THE PTOMAINES. 
233 
mixture of bases, which, on analysis, gave results corre- 
sponding to the three formula!: 
1 2 3 
(C5H14NOCl),PtCl4 (05HuNCl)aPtCl4 (C5H14NCl),PtCl4 
Formula No. 3 was the one accepted by Liebreich for 
neirrine, but, according to Baeyer, Liebreicii’s neurine 
salt is not simple, but is a mixture of Nos. 1 and 2. lie 
himself accepts formula No. 1 as the platinochloride of 
neurine, and distinctly states (Annal. <1. Chem. u. Pharm., 
142, 323, 1867) that neurine is in composition trimethyl- 
oxyethyl-ammonium hydroxide. And, according to him, 
choline from bile, and sinkaline from white mustard, appear 
to be identical with neurine. 
This nomenclature of Baeyer’s was at first adopted by 
Wurtz and others, who showed that the oxyethyl base 
was identical with choline and sinkaline. On that account 
Strecker, in 1868 (Annal., 148, 79), suggested the re- 
striction of the name choline to the oxyethyl base, and to 
reserve the name neurine for the base whose platinochloride 
is represented in No. 3, as originally was done by Lieb- 
reich. In 1869 Liebreich showed conclusively that 
pure protagon, when heated with baryta for twenty-four 
hours, yields a substance having the composition of the 
vinyl base: 
N(CH3)3.C2H3.OH. 
The platinochloride of this base crystallized in five-sided 
yellow plates, which, after a time, oil exposure to the air, 
became cloudy; on treatment now with water a portion 
dissolved, and the solution was found to contain the oxy- 
ethyl base. Furthermore, he observed that when the alco- 
holic extract of the brain, from which all the protagon had 
been removed, is treated with baryta, only the latter, the 
oxyethyl base, is obtained. Finally, in 1870, Wurtz 
abandoned the use of the term neurine to designate the 
oxyethyl base, and returned to the name choline, originally 
applied to the oxyethyl base by its discoverer, Strecker. 
Nevertheless, the confusion in the use of these two terms 234 
BACTERIAL POISONS. 
continued to exist, and even at the present time it is the 
cause of no little misunderstanding. Thus, Marino-Zuco 
(1885), in his excellent researches on the genesis of pto- 
maines, applies the term neurine, following Baeyer’s pre- 
cedent, to the oxyethyl base, C5H15N02, which is really 
choline, according to the proper nomenclature. 
We have gone somewhat at this point in detail into the 
history and the proper use of the terms neurine and choline 
because of the confusion which is sure to arise if the dis- 
tinction is not thoroughly borne in mind. The name 
neurine, then, should be used only to denote the vinyl base 
C5H13NO. It is trimethyl-vinyl-ammonium hydrate. On 
the other hand, choline is applied to the oxyethyl base 
c5ii,no2, which is trimethyl-oxyethyl-ammonium hydrate. 
Neurine has been obtained by Brieger (1883) in the 
putrefaction of horse, beef, and human flesh for five to six 
days in summer. It also occurs in the commercial, so-called 
“ neurine,” together with choline (Brieger, I., 34). Lier- 
reich obtained it in the decomposition of protagon by 
baryta. And Brieger (I., 60) also has isolated it along 
with choline from fresh human brains, by boiling with 
baryta; but has not obtained it by digesting the brains on 
the water-bath with two per cent, hydrochloric acid. It 
lias been found in putrid, and as result of this change 
poisonous, mushrooms (Berlinerblau, 1888). 
The genesis of neurine is still rather obscure, and it is 
to be hoped that future investigations may shed more light 
upon the mysterious production of this highly poisonous 
base. Its occurrence in the brain together with choline 
would seem to indicate that it is either derived from 
choline by the removal of water, or that it exists together 
with choline, partly replacing the latter in the molecule of 
protagon (lecithin), according to the hypothesis put for- 
ward by Lippmann (page 241). The question of its 
derivation from choline by withdrawal of a molecule of 
water has already been subjected to an interesting experi- 
mental discussion. Ch. Gram attempted to explain the 
production of neurine and other muscarine-like ptomaines 
as due to the dehydrating action of the acids employed in 235 
chemistry of the ptomaines. 
the methods of extraction, and, indeed, lie claimed to have 
converted choline platinochloride, by heating with hydro- 
chloric acid, into neurine. This statement has been dis- 
puted by Briefer, who showed that the platinochloride 
of choline, as well as the hydrochloride, may be heated 
with fifteen or thirty per cent., or even concentrated hydro- 
chloric acid, for six to eight hours on a water-bath, with- 
out any conversion whatever (III., 15). That neurine 
may be obtained from choline, at least by chemical pro- 
cesses, was shown by Baeyer, in 1866, who found that 
choline chloride, when heated with several times its volume 
of concentrated hydriodic acid and some red phosphorus, 
gave a compound C5H13NI2 which, on digestion with fresh, 
moist silver oxide, yielded a vinyl base identical with that 
previously obtained synthetically by Hofmann, and now 
known as neurine. Brieger has tried, unsuccessfully, to 
bring about this dehydration by the putrefaction of pure 
choline (I., 59). However, Schmidt and Weiss (1887) 
were more successful, and they found that choline, as well 
as the hydrochloride and lactate, is changed by the action 
of microorganisms into the strongly poisonous neurine. 
Their results are given in full uuder choline (see page 244.) 
From what has been said it is evident that neurine can 
only arise from choline, and this, as will be seen later, is 
derived from lecithin. 
Neurine is almost invariably accompanied by choline, 
from which, however, it can be readily separated by the 
difference in the solubilities of the platinochlorides. It 
occurs in the mercuric chloride precipitate (and in the 
filtrate), and from this it can be obtained, after removal of 
the mercury, by precipitating the solution of the mixed 
hydrochlorides in absolute alcohol by platinum chloride. 
The platinochlorides are then separated by recrystallization 
from water, since the neurine is difficultly soluble, while 
the choline salt is readily soluble. 
The free base possesses a strong alkaline reaction, and 
on contact with the fumes of hydrochloric acid it yields a 
cloud. According to Liebreich, the alkaline solution 
cannot be neutralized by passing through it carbonic acid. 236 
bacterial poisons. 
The Chloride, C5II12N.C1, is extremely poisonous, and 
crystallizes in fine hygroscopic needles. 
The Platinociiloride, (C5H12N.Cl)2PtCl4 (Pt = 33.60 
per cent.), is difficultly soluble in hot water, and crystallizes 
in beautiful, well-formed octahedra belonging to the regular 
system. No twin-crystals are observed. Sometimes the 
crystals contain water of crystallization, at other times they 
do not (Brieger, I., 33). According to Liebreich, it 
forms from an aqueous solution in five- or six-sided, heaped- 
up plates resembling urea nitrate, while from an alcoholic 
solution it forms needles, which on exposure to air become 
opaque, and are partially converted into the oxyethyl base 
—choline. 
The Aurochloride, C5H12N.C1.AuC13 (Au = 46.37 
per cent.), forms flat prisms, which are difficultly soluble 
in hot water (Brieger.) Dissolves easily, and can be 
purified by crystallization (Liebreich). 
Physiological Action.—Neurine is exceedingly poisonous, 
even in small doses, and in its action it strongly par- 
takes of the characteristic stamp of poisoning by muscarine. 
The injection of a few milligrammes into frogs produces 
in a short time a complete paralysis of the extremities, with 
deadening of reflex excitability. Respiration stops first, 
while the rate of heart-beat gradually decreases till, finally, 
stoppage in diastole takes place. The injection of atropine 
at this point does away with the effect of neurine, so that 
the heart begins to beat again. Previously atropinized 
frogs, as a rule, withstand the action of the poison. Im- 
mediately after the introduction of this substance there can 
be observed a distinct period of exaltation, which, however, 
soon gives way to the characteristic stage of depression seen 
in the progressive slowing of the rate of heart-beat. Of 
the warm-blooded animals, cats seem to be much more 
sensitive to its action than mice, rabbits, or guinea-pigs. 
The symptoms seen in rabbits are profuse moistening of the 
nasal cavities and upper lip, which is succeeded by an in- 
tensely profuse salivation; later on there is noticeable an 
abundant secretion from the nasal mucous membrane and 
from the eyes; the latter, however, ceases in a short time. CHEMISTRY of the ptomaines. 
237 
The movements of the heart and of respiration are at first 
quickened and strengthened, but before long the paralytic 
effects produce a constant slowing and weakening, till 
finally complete cessation of both movements results. The 
decided dyspnoea observed gradually alters its character, 
and just before death the respiration is irregular and super- 
ficial. The heart, as in frogs, continues to beat after the 
respiratory movements have ceased, until finally it stops in 
diastole. Direct application of concentrated solutions of 
the poison to the eyes produces almost always a contraction 
of the pupil, while a similar but less constant contraction 
is seen when it is injected. The peristaltic action of the 
intestines is heightened to such an extent that continual 
evacuation takes place. Just before death, violent clonic 
convulsions occur. Atropine possesses a strong antagonistic 
action toward neurine, and the injection of even a small 
quantity is sufficient to dispel the symptoms just de- 
scribed. 
Choline, C5H15N02 = C2H4OH.N(CH3)3.OH. — This 
base is identical with the sinkaline of yon Babo, the bili- 
nenrine of Liebreich, and the nenrine of Baeyer, 
Marino-Zuco, and others. According to Schmiedeberg 
and Harnack, it is identical with Letellier’s amanitine 
(agaricine), to which they assign, however, the formula 
(CH3)3N.(CHOH.CH3)OH. Choline was first prepared, 
and so named, by Strecker, in 1862, by treating hog-bile 
with hydrochloric acid. It was prepared synthetically by 
Wurtz (1868) by direct union of ethylene chlorhydrine 
and trimethylamine. 'File reaction that takes place can be 
represented by the equation : 
f OH ) cr3! 
cT + N= cil:i NCI 
3 ’ c;ii(.oh j 
Baeyer (1866) obtained it by boiling an alcoholic extract 
of the brain with baryta water; and Liebreich, in 1869, 
showed that if the alcoholic extract, from which all the 238 
BACTERIAL POISONS. 
protagon liad been removed, be thus treated, only choline is 
formed, whereas pure protagon, on heating with baryta, 
yields neurinc. Jt has been obtained from the yelk of 
eggs ; from bile; from fresh brains (Brieger) ; from fresh 
eggs, blood, lungs, and hearts, and from lecithin (Marino- 
Xijco) ; from human placenta (Boehm); from the eye; 
from commercial neurine (Brieger) ; from fresh as well as 
decomposing internal organs of the cadaver (Brieger, 
1885); from herring-brine and decomposing pike, three 
days in midsummer (Bocklisch). It has also been isolated 
from cultures of vibrio proteus (Bocklisoii), and of comma 
bacillus (Brieger). Ehrenberg (1887) found it in 
poisonous sausage, and, by growing a bacillus obtained from 
this, on liver. 
Not only has choline been met with in the animal tissues, 
but it has also been observed within the last few years to be 
very widely distributed in the vegetable kingdom, especi- 
ally so in fatty seeds. Thus, it has been found (Harnack, 
1876) accompanying muscarine, in toadstool (Agaricus mus- 
carius); in hops, and lienee in beer (Griess and Harrow ; 
in the seeds of Trigonella, in Indian hemp, areea- and earth- 
nuts, hemp seeds and lentils (Jahns); in the seeds of white 
mustard, as a glycoside (von Babo); in ergot (Brieger); 
in the germs of pumpkins and lupines (Schulze, Zeitsokr. 
f. Physiol. Chem., 11, 365); in beech-nuts and morels (Hel- 
vetia eseulenta, Boletus lusidus, Amanita pantherina, Bohm) ; 
in flores sambuci (elder), and extracts of belladonna, hyos- 
cyamus, ipecacuanha root and Acorus calamus (Kunz), 
and Scopolia Japonica (Schmidt and Henschke) ; in the 
sprouts and cotyledons of Sqja beans (Schulze, 1888), in 
the fiit from hog’s bean, vetch, peas and lupines (Jacob- 
son, 1889); from the lecithin of lupine seeds (Schulze and 
Steiger) ; and in Chekeu leaves (Myrtus cheken, Weifs). 
According to Lirrmann {Ber. 20, 3206), it is present, to- 
gether with betaine, in the molasses from beet-root sugar. 
Choline (Ritthausen) and betaine (Bohm) exist together 
in cotton-seeds; hence, choline occurs in the press-cakes from 
cotton seeds (Bohm). According to Schulze, and also 
Ritthausen, choline occurs with betaine and another CHEMISTRY OF THE PTOMAINES. 
239 
base in the seed of the vetch, and in peas with a base re- 
sembling betaine. The two bases have also been found 
together in Scopolia atropoides by Siebert. 
Choline may readily be prepared, after the method of 
Diakonow, from the yelk of eggs. These are extracted 
with ether, then with alcohol, and the extracts thus ob- 
tained evaporated, when the resulting residues are boiled 
with baryta for one hour. The filtrate, after the removal 
of the barium by carbonic acid, is evaporated and the 
residue is abstracted with absolute alcohol. The alcoholic 
solution is now precipitated with platinum chloride. 
Brieger (II., 55) has presented a method which is much 
simpler in its details and obviates the use of the expen- 
sive platinum chloride. The tissues rich in lecithin, as 
yelk of egg, brain, etc., are heated with concentrated 
hydrochloric acid for some hours on the water-bath. The 
insoluble residue is filtered off, and the filtrate, after neu- 
tralization of the excess of free acid with carbonate of 
sodium, is evaporated. The residue is extracted with 
alcohol, and the alcoholic solution is precipitated with 
alcoholic mercuric chloride. The precipitate thus obtained 
on recrystallization several times from a large quantity of 
boiling water, yields the pure double salt of choline. 
If desirable, it can be made from pure lecithin, best pre- 
pared according to Gilson’s method. Yelk of eggs is 
repeatedly shaken up with ether until the latter is colored 
only a faint yellow; the ether solution then distilled, the 
residue taken up in petroleum ether and filtered. The 
filtrate, in a separatory funnel, is well shaken with 75 
per cent, alcohol, and tin’s is repeated several times with 
fresh alcohol. The alcoholic extracts are combined, allowed 
to stand for some time, then filtered and subjected to dis- 
tillation to remove traces of petroleum ether. The solu- 
tion is now set aside in a cool place for several days; the 
precipitate which forms consists of cholesterine, etc., and 
a little lecithin. The alcoholic solution is filtered by de- 
cantation, then decolored by boiling with bone-black; 
rapidly evaporated at 50-60° to a syrupy consistency. 
This residue is extracted with ether, the solution filtered 240 
BACTERIAL poisons. 
and evaporated. The lecithin thus obtained is almost per- 
fectly pure, but contains traces of cholesterine. To com- 
pletely purify it, it can be dissolved in as little absolute 
alcohol as possible, and set aside to reprecipitate in the 
cold, —5 to 15°. 
In regard to the genesis of choline the preponderance of 
testimony goes to show that it is derived from the decom- 
position of lecithin, which, according to the researches of 
Diakonow and others, is one of the most widely distributed 
compounds, occurring in greater or less quantity in all of 
the animal tissues. Lecithin, which is a complex esther 
(Strecker, Hundeshagen, Gilson), decomposes under 
the action of acids and alkalies into a base (choline) 
glycerin, phosphoric acid, and fatty acids (stearic, oleic, 
palmitic, etc.). Gilson has shown that dilute sulphuric 
acid slowly decomposes lecithin, forming choline, which, 
after a few days, disappears; on the other hand, sodium 
hydrate, in even 1 per cent, solution, rapidly decomposes 
it. This change is undoubtedly accomplished in a similar 
manner through the agency of bacteria. Brieger (II., 
17) is inclined to believe that choline exists preformed in 
the various tissues, inasmuch as he has been unable to ob- 
tain it from the brain, which is rich in lecithin, by boiling 
with 2 per cent, hydrochloric acid. (See Schulze, page 
242.) Prolonged heating with concentrated hydrochloric 
acid was necessary in order to obtain any choline from the 
brain. This result of Brieger’s is somewhat at variance 
with that of Marino-Zuco (see Relazione, etc., pages 29, 
30, and 38), who obtained from 25 grammes of lecithin, by 
the method of Stas, a small quantity of the aurochloride 
of a base, while from a similar amount he obtained more 
relevant quantities by the method of Dragendoeff. 
The occurrence of choline in the vegetable kingdom 
would be inexplicable to us at present were it not that 
we now know of the existence of lecithin-like bodies in 
plants, from the decomposition of which substantially the 
same products are obtained as from the lecithin obtained 
from the animal tissues. The existence of such a body in 
plants was first predicted by Scheirler in 1870, who was chemistry of the ptomaines. 
241 
led to this conclusion in his celebrated study of beet-root 
sugar, because of the presence of oleic acid, glycerin, phos- 
phoric acid, and betaine, as well as cholesterin, in the beet- 
root extracts. This hypothesis was confirmed by Hoppe- 
Seyler, who, in 1879, found a lecithin substance in yeast. 
Schulze found a similar compound in the cotyledons of 
lupine, while Jacobson observed its presence in mustard- 
seeds, in fenugreek-seeds, in maize and wheat, in the fat 
from beans, peas, vetch, and lupines. Heckel showed its 
presence in globularia, and Lippmann has found it in beet- 
root. According to Hoppe-Seyler, this lecithin-like sub- 
stance exists in all vegetable cells undergoing development. 
Schulze and Likiernik (1891) were the first to prepare 
lecithin in a pure condition from plants. It was found to 
possess the same properties and yield the same decomposi- 
tion-products as lecithin from animal tissues. Up to the 
present time lecithin has always been supposed to contain a 
radical, which gives rise to choline on saponification, as an 
essential component, while on the other hand the fatty 
acids entering its molecule are well known to be replaceable 
by one another. Thus we may have a di-stearine lecithin 
as well as a di-oleine lecithin. The existence of several 
lecithins in the yelk of eggs has been recognized for some 
time, and according to Schulze and Likiernik this is 
also true of the lecithins in plants. Recent observations 
of Lippmann (Ber. 20, 3206) show that the above basic 
radical, hitherto regarded as constant in lecithin, may pos- 
sibly be capable of replacement by other similar radicals. 
He found on saponifying with baryta two different speci- 
mens of lecithin, both obtained from beet-root, that while 
one of them yielded oleic acid, glycerin, phosphoric acid, 
and betaine; the other lecithin gave oleic acid (and some 
other fatty acids), glycerin, phosphoric acid, and choline, 
with no betaine—at least not in isolable quantity. This 
remarkable difference has led Lippmann to suggest an ex- 
planation which, while it may not be the correct one, never- 
theless possesses a high degree of probability. According 
to him, the lecithin molecule may contain interchange- 
able basic radicals in the same manner that it contains 242 
BACTERIAL POISONS. 
interchangeable acid radicals. This view is supported not 
only in the case of beet-root, where choline and betaine 
exist together, but the same two bases have been observed 
in cotton-seeds. A similar coexistence was observed in the 
toad-stool (Agaricus muscarius), in which choline and mus- 
carine were found. And, lastly, the same condition holds 
true probably for mytilotoxine and betaine, which were 
shown to be present together in poisonous mussels. 
Lecithin cannot always be regarded as the source of 
choline in plants, since this base is known to occur as a 
glucoside in the seeds of white mustard. The sinapin de- 
composes according to the equation : 
Ci6H33N05 + 2H20 = C5H15N02 + CuHl205. 
Sinapin. Choline. Sinapic Acid. 
According to Schulze (1891) the choline which is iso- 
lated from pea- and vetch-seeds exists preformed in the 
seeds, and does not result from lecithin by the process of 
extraction. This is also probably true with reference to 
cottonseed-cake. The condition in which betaine exists is 
not determined. 
The protoplasm itself is another possible source of choline 
as well as of other nitrogenous bases, as xanthine, etc. We 
know from Diiechsel’s brilliant investigation (1890) that 
casein on treatment with hydrochloric acid and stannous 
chloride yields ammonia, amido acids, and organic bases— 
lysatine, C6H13N302, and lysatinine, C6HuN30 — homo- 
logues of creatine, C4II9N302, and creatinine, C4II7lSr30. 
From lysatinine urea can be readily obtained by treatment 
with baryta. Subsequently, Siegfried (1891) showed that 
vegetable protoplasm (conglutin from lupine) when treated 
in the same way yields similar products. Later, Schulze 
demonstrated that the base, arginine, C6H14N402, is formed 
in lupine sprouts at the expense of the proteids present, and 
he pointed out that this base is probably related to lysatine, 
from which it differs only by NH (sec next chapter). 243 
CHEMISTRY OF THE PTOMAINES. 
Decompositions of Choline.—Baeyer (1866) suc- 
ceeded in converting choline into neurine by a purely 
chemical process. This was accomplished by heating 
choline chloride with concentrated hydriodic acid and 
red phosphorus in a sealed tube at 120°-150°, whereby 
the compound C3HI3NT2 was formed. The iod-iodide of 
choline thus obtained, on treatment with moist silver oxide, 
gave a base whose platiuoehloride corresponded to the 
formula (C5H12NCl)2PtCl4-TH20. This double salt, ac- 
cording to Baeyer, is readily soluble in water, and gives 
reactions similar to choline. Although Baeyer is em- 
phatic in his assertion that this is the vinyl compound 
(neurine) formed from the oxy-ethyl base (choline), yet it 
seems that there is room for doubt in regard to the 
interpretation of his results. Thus neurine platiuoehloride 
is difficultly soluble in water, contrary to the behavior of 
the platiuoehloride obtained by him. On the other hand, 
choline platiuoehloride is easily soluble in water, and it 
would seem, therefore, that Baeyer has not converted 
choline into neurine, but rather has regenerated choline 
from its iod-iodide. If such were the case, we would ex- 
pect that the iod-iodide of neurine, C5II13NI2, which has 
the same composition as the corresponding derivative of 
choline, would yield, on treatment with silver oxide, the 
oxy-ethyl base. Baeyer has apparently not been able to 
effect this change, since he holds that the vinyl base may 
be prepared from the oxy-ethyl, but that the reverse, the 
preparation of the oxy-ethyl base from the vinyl compound, 
cannot be accomplished. 
Whether.the change described by Baeyer takes place or 
not, it is, nevertheless, certain that choline does not readily 
give up a molecule of water and thus become converted 
into neurine. Ch. Gram announced, in 1886, that choline 
chloride and lactate on heating on the water-bath de- 
compose, and that this conversion into the vinyl base was 
complete when the aqueous hydrochloric acid solution of 
choline platinochloride was heated for five or six hours on 
the water-bath. In this way Gram endeavored to explain 
the formation as due to the action of acids upon choline, 244 
BACTERIAL POISONS. 
but Brieger has shown that the platinum salt of choline, 
as well as its hydrochloride, can be heated with fifteen or 
thirty per cent., or even concentrated, hydrochloric acid for 
six or eight hours without undergoing any change into 
neurine, thus disproving the results obtained by Gram. 
E. Schmidt has confirmed Brieger’,s observations in 
regard to the resistance of choliue to decomposition by 
acids, but he has gone further, and has shown that what 
the action of acids has failed to do is readily accomplished 
through the agency of bacteria. He found that choline 
chloride, when allowed to stand with hav infusion, or with 
dilute blood for fourteen days at 30°—35°, it almost entirely 
decomposed, yielding large quantities of trimethylamine 
and a base, the platinochloride of which resembles in form 
and solubility the double salt of neurine, and possesses a 
similar physiological action. Choline lactate in hay infu- 
sion developed an odor of trimethylamine in twelve hours, 
but at the end of fourteen days a good deal of choline was 
still present. In this case no neurine was present, but 
instead a homologous base was found, which can be obtained 
synthetically by the action of trimethylamine on allyl 
bromide. According to Meyer, of Marburg, this base 
does not possess the muscarine-like action of neurine, but 
resembles more closely pilocarpine. 
Brieger (I., 59) had unsuccessfully tried to transform 
choline into neurine by putrefaction. He observed that the 
choline decomposed with extreme slowness, even when the 
putrefaction was carried on at a higher temperature, yield- 
ing only trimethylamine. Wurtz (1868) showed that 
dilute solutions of free choline can be heated to boiling 
without any perceptible decomposition. Concentrated 
solutions, however, decompose with the formation of tri- 
methylamine and glycol, C2H4(OII)2 (see page 190). The 
decomposition of choliue was studied somewhat by 
Mautiiner (1873), who confirmed Wurtz’s observation 
that choline was scarcely decomposed by boiling water, and 
he showed that when exposed to the action of decomposing 
blood it yielded trimethylamine. The results obtained by 
K. Hasebroek (Zeitsehrift f. Physiol. Chem., 12,151, 1888) CHEMIS T R Y OF T LI E PTOMAINES. 
245 
deserve special mention at this place. lie carried on the 
putrefaction of very dilute solutions of the chloride of 
choline in the presence of little or no oxygen in Hoppe- 
Seyler fermentation flasks. Sewer slime, because of its 
strong fermentative properties, was used to induce the 
putrefaction, and calcium carbonate was added to neu- 
tralize any acidity that might develop during the fermen- 
tation. 
The fermentation, as shown by the evolution of gases, 
lasted for about three months. The total quantity of gas 
given off’ was about one litre from 1.17 grammes choline 
chloride. The gases consisted almost entirely of carbonic 
acid and marsh gas. No hydrogen was evolved. When 
the fermentation ceased the flask was opened and several 
cubic centimetres of the almost neutral clear liquid were 
injected under the skin of a rabbit without producing the 
least effect. 
This liquid distilled with alkali gave methylainine 
and ammonia. What is remarkable about this experiment 
was the total absence of the higher amines—as, for instance, 
trimethylamine, which has been observed so many times as 
a decomposition-product of choline. The absence of any 
poisonous base, as neurine, was probably largely connected 
with the absence of oxygen. 
Free choline ordinarily forms a strongly alkaline syrup 
which combines readily with acids to form salts, most of 
which arc deliquescent. By oxidation it is converted into 
betaine (see page 249), and on treatment with concentrated 
nitric acid it gives rise to muscarine (see page 251). These 
reactions can be represented by the equations : 
CH2OH C<oh 
I I 
ch2 +o2= ch2 +h2o. 
l(CH3)3.OH N(CH3)3.OH 
Choline. Betaine. 246 
bacterial poisons. 
CH,OH CII2OH 
I I' 
CH2 + O = CHOH 
I ‘ I 
N(CH3)3.OII N(CH3)3.OH. 
Muscarine. 
By the action of dilute nitric acid choline is converted 
into a base the platinochloride of which is efflorescent and 
corresponds to the formula (C4H10N2O3Cl)PtCl4 + 2H20 
(SCHMIEDEBERG and HaRNACk). 
According to Mautiiner, choline resembles the caustic 
alkalies in its action. Although putrefying blood decom- 
poses it into trimethylamiue, yet, when present in the pro- 
portion of 1.4 percent., it is said to arrest putrefaction. 
A 1 to 2 per cent, solution is said to dissolve fibrin or 
coagulated albumin on boiling. 
The free base, as well as the carbonate, is dimorphous 
and forms thin plates or long needles. 
The Chloride, C5H14NO.CI, is easily soluble in water 
and in absolute alcohol (separation from neuridine hydro- 
chloride). It crystallizes over sulphuric acid to needles 
which readily deliquesce in the air. 
The Platinochloride, (C5H14NO.Cl)aPtCl4 (Pt = 
81.64 per cent.), presents an interesting case of trimorph- 
ism. It crystallizes in monoclinic plates (Rinne) which 
are easily soluble in water, insoluble in alcohol; also in 
characteristic superposed plates, sometimes in the form of 
orange-red flat prisms (Brieger). From a warm saturated 
solution containing 15 per cent, alcohol it crystallizes in 
yellow regular octahedra containing one molecule of water 
of crystallization (JaHns); from aqueous solution on slow 
evaporation it forms plates, clinorhombic prisms, or needles 
(Hoppe-Seyler) which are anhydrous. When rapidly 
crystallized it forms prisms (IIundeshagen, Jahns, 
Schulze) ; and if the solution is concentrated the prisms 
are very thin, almost needles. According to Schulze, it 
sometimes forms beautiful orange-red, chiefly six-sided 
plates. Jahns maintains that the plates and prisms be- chemistry of the ptomaines. 
247 
long to the same system ; while HundeSHAGEN holds that 
they are distinct. Instead of the salt presenting an in- 
stance of trimorphism as first stated by Hundesiiagen, it 
would seem that but two forms occur—anhydrous mono- 
clinic and octahedra with oue molecule of water of crystal- 
lization. It contains always more or less water of crystal- 
lization which it does not give up completely over sulphuric 
acid, but only at 110° (Brieger). The natural platino- 
chloride becomes strongly electric on rubbing, whereas the 
synthetic choline double salt does not become electric. It 
melts at 225° with effervescence (Jahns). 
The Aurochloride, C5H14NO.C1.AuC13 (Au = 44.48 
per cent.), is crystalline and is difficultly soluble in cold 
water, but can be recrystallized from hot water or from 
boiling alcohol. It forms prisms, or gold-yellow long 
needles, which are very easily soluble in hot water and 
alcohol (Lippmann). It can be separated from neuridine 
aurochloride by its solubility in water (Brieger). On 
heating, the gold salt melts to a brown liquid (Schulze) 
and decomposes at 264°. 
The Mercurociiloetde, C5IIuNO.C1.6HgCl2, is ex- 
tremely difficulty soluble even in hot water. On this 
account the mercury salt is very convenient for the separa- 
tion of choline from accompanying bases. 
The Bicrate, C5H14N0.0C6H2(NO2)3, forms long, broad 
needles which arc more easily .soluble than neuridine'picrate, 
and hence can be separated by recrystallization. It is more 
easily soluble in alcohol than in water. 
Physiological Action of Choline.—Choline was regarded 
for a long time as physiologically inert, but this belief 
was set aside by Gaehtgens (1870), who showed that, 
when given in large quantity, it possessed a toxic action. 
This observation of Gaehtgens has since been con- 
firmed by Glause and Luciisinger, Brieger, and 
Boehm. The chloride of choline produces in animals the 
same muscarine-like symptoms of poisoning as are devel- 
oped by the vinyl base neurine, the only difference lies in 
the intensity of the action. In order to bring about a 
physiological disturbance, choline must be given in rela- 248 
BACTERIAL POISONS. 
tively large doses. Thus, Brieger found it necessary to 
give about 0.1 gramme of choline chloride hypodermi- 
cally to a one kilogramme rabbit in order to bring out the 
same effects as are obtained by the injection of 0.005 
gramme of the neurine salt. He also found that the 
fatal dose for a one-kilogramme rabbit was about 0.5 
gramme, which is about ten times as large as the fatal 
close of neurine chloride. Boehm observed that doses of 
0.025-0.1 gramme produced in frogs general paralysis, 
which, in a short time, leads to death or recovery; and 
that in its curara-like paralyzing action, choline resembles 
artificial muscarine, although the latter is about live 
hundred times stronger. Atropine, as in the case of 
neurine and muscarine, antagonizes the action of choline. 
Thus, 0.05 gramme of the chloride produced in a frog in 
one hour diastolic stoppage of the heart. This condition 
was removed by the injection of 0 001 gramme of atropine, 
the heart-beat rising to the normal in about fourteen min- 
utes; 0.05 gramme of choline chloride, given subcutane- 
ously to a rabbit (1250 grammes) produced salivation, which 
lasted but a short time, and did not affect the heart-beat 
and respiration; 0.10 gramme was necessary to bring out 
all the symptoms; 0.05 gramme, given to guinea-pigs, had 
no effect whatever. 
Betaine (Oxyneurine), C5H13N03.—This base lias 
been well known for some time, beeause of its occurrence 
in the vegetable kingdom. Thus, it is present in cotton- 
seed (Boehm, Ritthausen and Weger); in beet-root juice 
(Beta vulgaris), and hence in beet-root molasses (Schei- 
BLER, 1806). It occurs also in cattle-turnip and Lycium 
barbarum ; and is found with choline and another base in 
vetch-seeds; in peas a base similar to betaine exists 
(Schulze). With choline it occurs in Scopolia atropoides 
(Siebert). It does not exist in these substances as such, 
but is formed from a more complex substance by the action 
of hydrochloric acid or baryta (Liebreicii). In this respect 
it resembles choline, neurine, and probably muscarine. 
Quite recently, Lippmann (1887) has obtained a lecithin- CHEMISTRY OF THE PTOMAINES. 
249 
like body from sugar-beet, which, on heating with baryta 
gave oleic acid, glycerin, and phosphoric acid (glycerin- 
phosphoric acid), and betaine. Betaine, however, does not 
seem to be a constant constituent, inasmuch as on one occa- 
sion he obtained chiefly choline, and little or no betaine. 
These two bases also occur together in cotton-seed, and 
this fact has led Scheibler to the conclusion that it is 
no mere chance. Lecithin, as is well known, may con- 
tain variable acid constituents (oleic, stearic, palmitic, etc.), 
and reasoning on this fact, and on the results of his experi- 
ments, Lippmann has been led to suppose that it may also 
contain different bases in variable proportions. 
It has been obtained from human urine (Liebreicji, 
1809), and from poisonous and non-poisonous mussel, but 
not from putrid mussel (Brieger, 1885, III., 76). The 
method for its separation from mussel is described on page 
255. 
Betaine may be obtained synthetically in several ways : 
(1) by oxidation of choline with potassium permanganate; 
(2) by the action of methyl iodide on glycocoll; (3) by 
treating monochloracetic acid with trimethylamine. The 
last two methods are of value as indicating the constitution 
of betaine, and the changes which take place can be repre- 
sented by the equations: 
NH2 N(CH3)3I 
c!h2 + 3CU3I = CH2 + 2HI. 
I I 
co2h co2ti 
Glycocoll. 
Betaine Iodide. 
N(CH3)3C1 
CII2C1 | 
| “ + N(CH3)3 = oh2 
coji I 
co2h. 
Monochloracetic Acid. 
From the formulae of the salts of betaine it is evident 250 
BACTERIAL POISONS. 
that betaine has properly the composition C5H13N03, which 
is expressed by the structural formula : 
N(CH3)3OH 
I 
cir2 
I 
00,11. 
The free base is, however, readily converted into the 
anhydride, CflHnN02, trimethyl glycocoll; the structural 
formula of which is : 
CH2-N(CII3)3 
I I 
CO —o. 
Betaine is ordinarily regarded as crystallizing with one 
molecule of water, and the composition is expressed by the 
formula: C5HuN02+H20 (= OH.N(CH3)3.CII2.CG2H). 
It loses this water of crystallization by heating at 100°, or 
on standing over sulphuric acid, forming an anhydride of 
the formula already given. Liebreich claims that free 
betaine possesses the formula C5HUN()2, because it yields a 
compound having the composition (C5lIuN02)ZnCl2. The 
free base separates from alcohol in large crystals which deli- 
quesce on exposure to the air. As obtained by Brieger 
from the hydrochloride by treatment with moist silver 
oxide, it possessed a sweetish taste and neutral reaction. 
When distilled with potassium hydrate, it yields trimethyl- 
amine and other bases, among which a base of the formula 
C8IIi7N05 occurs in the largest quantity. 
The Chloride, C5II12N02.C1, forms beautiful crystals, 
monoclinic plates, which are permanent in the air, and this 
can be made use of to effect a separation from the choline 
salt, which is deliquescent. It is insoluble in absolute 
alcohol. This fact can be made use of in their separation 
(Lippmann). It can, moreover, be easily separated from 
other bases by its aurocldoride, which is easily soluble. If 
a little potassio-mercuric iodide is added to a solution of 
the chloride, there forms a light-yellow or whitish oily 251 
CHEMISTRY of the ptomaines. 
precipitate, which is soluble in excess, but on rubbing the 
sides of the tube with a glass rod it reappears as yellow 
needles. This is said to be a characteric test (Brieger, 
Schulze, 1891). 
The Aurochloride, C5II12N02.C1.AuC13 (Au = 43.12 
per cent.), forms magnificent cholesterin-like plates, and is 
easily soluble (Brieger). The aurochloride from sugar- 
beet is said to crystallize in needles or plates, and to be 
difficultly soluble in cold water (Scheibler, Lippmann). 
The double salt of the ptomaine melts at 209°, and in this 
it coincides with that obtained from beet-sugar, as well as 
with that of the synthetically prepared base (Brieger). 
The platinochloride is yellow and crystalline. 
Betaine is not poisonous. 
Muscarine, C5II15N03 = C5H13N02 + II20, the well- 
known toxic principle which Schmiedeberg obtained 
from poisonous mushroom (Agaricus muscarius), has been 
obtained also by Brieger in 1885 (I., 48) from haddock 
which had been allowed to decompose for five days. The 
process by which its isolation was effected is described on 
page 258. This base is specially interesting, because of the 
relation it bears to choline, for Schmiedeberg has shown 
that it is formed when choline, or, better still, the platino- 
chloride, is oxidized by concentrated nitric acid. It is 
barely possible that Brieger’s base is distinct from 
Schmiedererg’s ; nevertheless, it closely resembles it and 
apparently is identical. 
The Chloride, C5II14N02.C1, is obtained on the decom- 
position of the platinochloride with hydrogen sulphide, as 
a syrupy residue, which, under the desiccator, shows a 
tendency to gradually crystallize. It is deliquescent 
(Harnack). 
The Platinooiiloride, (C5H14N02.Cl)2PtCl4 (Pt = 
30.08 percent.), forms as a crystalline deposit of octahedra, 
which are difficultly soluble in water. They lose their 
water of crystallization (2H20) only by means of strong 
heating. 
The Aurochloride, C6II14N02.C1.AuC13, crystallizes 252 
BACTERIAL POISONS. 
in needles, and is difficultly soluble in water, more so than 
the choline double salt (Harnack). 
Physiological Action.—Small (loses of this ptomaine 
induce in frogs total paralysis, with stoppage of the heart 
in diastole, and this action is antagonized by subsequent 
injection of atropine, as well as in the case of previously 
atropinized frogs. Very small doses produce in rabbits 
profuse salivation and lachrymation, contraction of the 
pupil, profuse diarrhoea, and passage of urine and semen ; 
finally, the animal dies in convulsions, which, however, are 
only of short duration. 
Constitution of the Members of the Choline 
Group.—The structure of choline was clearly demon- 
strated by Wurtz, who accomplished the synthesis of this 
base by treatment of ethylene chlorhydrine with trimethyl- 
amine. This same method can be applied to the synthe-is 
of betaine and neurine by using monochloracetic acid and 
vinylbromide instead of ethylene chlorhydrine. The struc- 
tural formulae which can be deduced from these reactions 
are as follows: 
Cn2OH CH2 C02H CH2OH 
I II I I 
ch2 on ch2 choh 
I I I I 
N(( TI3)3.01 r N(CH3)3.( >1 T N(CH3)3.OII N(CH3)3.( )II 
Ciioi.ine. Neurine. Betaine. Muscarine. 
The formula) of betaine and muscarine are ordinarily given 
as the anhydrides, but there can be no doubt that the free 
bases possess the structure indicated above. All these bases, 
since they can be prepared from choline, may also be con- 
sidered as oxidation-products of trimethyl-ethyl-ammonium 
hydrate : 
CHS 
I ■ 
cit2 
I 
N(CII3)3.OH. CHEMISTRY OF THE PTOMAINES. 
253 
Mydatoxine, C6H13N02.—This base was obtained by 
Brieger in 1886 (III., 25, 32) from several hundred 
pounds of human internal organs which were allowed to 
stand in closed but spacious wooden barrels for four months, 
at a temperature varying from —9° to +5°. He obtained 
much larger quantities of it, however, from horseflesh which 
had putrefied under the same conditions. In the process 
of extraction it is found in the mercuric chloride precipitate 
together with cadaverine, putrescine, and another base, 
C7H17N02. It can be isolated from this mixture by recrys- 
tallizing the mercury salts, which removes the cadaverine, 
because of its difficult solubility in water, and decomposing 
the soluble mercury salts by hydrogen sulphide. The 
filtrate freed from mercury is now evaporated to dryness 
and the residue repeatedly extracted with absolute alcohol, 
in order to remove putrescine hydrochloride, which is 
insoluble. The alcoholic solution, after standing some time 
to permit complete separation of any dissolved putrescine, 
is then evaporated to dryness and taken up with water. 
This solution gives, on the addition of gold chloride, a pre- 
cipitate of the aurochloride of the base C7H17N02. The 
filtrate from this precipitate, containing the mydatoxine, is 
treated with hydrogen sulphide to remove the gold, and 
then evaporated to dryness. The colorless, syrupy hydro- 
chloride thus obtained forms with platinum chloride a 
double salt which is readily soluble in water, and can be 
purified by repeated recrystallizations from absolute alcohol 
containing some hydrochloric acid. 
The name mydatoxine is derived from /uvddo, to putrefy. 
The free base is obtained from the hydrochloride by treat- 
ment with moist, freshly precipitated silver oxide, as a 
strongly alkaline syrup, which solidifies in vacuo to plates. 
It is insoluble in alcohol, ether, etc. It does not distil 
without decomposition. It is isomeric with the base, 
06H13NO2, obtained by Brieger in 1888 from tetanus 
cultures. 
The Hydrochloride, C6H13H02.HC1, is a colorless, 
deliquescent syrup which does not form any double salt 
with gold chloride. With platinum chloride it gives an 254 
BACTERIAL POISON'S. 
easily soluble salt. Otherwise it combines only with plios- 
phomolybdic acid, with which it forms cubes. Ferric 
chloride and potassium ferricyanide yield, after a time, 
Berlin-blue. It is readily soluble in alcohol. 
The Platinochloride, (C6II13N02.IICl)2PtCI4 (Pt — 
29.00 per cent.), melts at 193°, with decomposition. It 
crystallizes in plates which are extremely soluble in water. 
It can be readily recrystallized from absolute alcohol acidu- 
lated with hydrochloric acid. The mercury salt is readily 
soluble in water. 
The exact formula of this base, of mytilotoxine, and 
some other bases, cannot be considered to be permanently 
settled, inasmuch as the formula of the hydrochloride, 
C6H13N02.HC1, as deduced from the analysis of the 
platinum double salt, may equally apply to the base 
C6HI4N02.OII as to the base Cgil13NC)2. If the first 
formula is correct, then mydatoxiue is a homologue of 
betaine, and its structure would be expressed by (1). 
(3) • 
COJI 
I 
ch2 
I 
CII2 
I 
N(CII3)3OH 
(2) 
C^° 
I Ml 
ci i 
II 
CII 
I 
N(CII3)3OII. 
The second formula would seem to correspond to an uu- 
saturated aldehyde of the choline group and its structure 
may be indicated by (2). 
This ptomaine, although it possesses toxic properties, is 
not, however, a strong poison. Its action is the same as that 
of the base C7II17N()2 (see page 262), with which it is associ- 
ated, except that the symptoms of poisoning develop slower, 
so that the death of a guinea-pig does not take place for 
about twelve hours. White mice are very susceptible to the 
action of these two poisons. A short time after the injec- 
tion of even small doses they are taken with convulsions CHEMISTRY OF THE PTOMAINES. 
255 
which come on in paroxysms. The eyeballs roll upward. 
Laehrymatiou, diarrhoea, and dyspnoea come on, and the 
mice die within a short time. 
A Base (?), C6TI13N02, an isomer of the preceding, was 
obtained by Brieger in 1888 from tetanus cultures. It 
is not poisonous—distinction from mydatoxine. It proba- 
bly is an amido-acid. The platinochloride crystallizes in 
plates, is easily soluble in water and in alcohol, and melts 
at 1!>7° with decomposition (see page 267). 
Mytilotoxine, C6II15N02, is the specific poison of toxic 
mussel (Mytilus edulis), from which it was obtained by 
Brieger in 1885(111., 76). This poison is formed during 
the life of the animal under certain conditions which have 
been thoroughly studied by Schmidtmann, Virchow, and 
others (see p. 40). Brieger obtained the poison by extract- 
ing toxic mussel with acidulous water, and evaporating this 
solution to a syrupy consistency. The residue was thor- 
oughly extracted with alcohol, and this solution was treated 
with lead acetate, in order to remove mucilaginous sub- 
stances. The filtrate was then evaporated, and the residue 
extracted with alcohol. Any lead that had dissolved was 
removed by hydrogen sulphide. The alcohol was expelled, 
and the resulting syrup was taken up with water and 
decolored by boiling with animal charcoal. The clear solu- 
tion was now neutralized with sodium carbonate, acidulated 
with nitric acid, and precipitated with phospliomolybdie 
acid. The precipitate was decomposed by warming with 
neutral lead acetate, and the resulting filtrate, after the 
removal of the lead by hydrogen sulphide, was acidulated 
with hydrochloric acid and evaporated to dryness. The 
residue was extracted with absolute alcohol, whereby 
betaine, on account of its insolubility, is removed, and the 
alcoholic solution was precipitated by alcoholic mercuric 
chloride. The mercury precipitate is repeatedly recrystal- 
lized from water, and the poison is obtained as an easily 
soluble double salt. 
The free base as obtained by the addition of alkali to 256 
BACTERIAL POISONS. 
the hydrochloride possesses a disagreeable odor which dis- 
appears on exposure to air, and the substance ceases to pos- 
sess poisonous properties. B rieger has proposed the 
application of this test for the recognition of poisonous 
mussel; on treatment of these with alkali the characteristic 
odor is developed. Mytilotoxine is also destroyed on dis- 
tillation with potassium hydrate and in the distillate there 
is found an aromatic non-poisonons product and trimethyl- 
amine. The free base, therefore, does not exist by itself 
for any length of time, but soon becomes converted into an 
inert substance. H. Salkowskt has also shown that it is 
destroyed on boiling with potassium carbonate, whereas 
its hydrochloric acid solution can be evaporated to dry- 
ness and heated to 110° without destroying its poisonous 
property. 
The Hydrochloride, C6II]5N02.IIC1, prepared from 
the aurochloride, crystallizes in tetrahedra. It is extremely 
poisonous and according to Brieger produces exactly the 
same symptoms which have beeu observed by Schmidt- 
mann in persons who have partaken of poisonous mussels 
(see page 38). On standing, however, the pure hydro- 
chloride gradually becomes dark and decomposes with loss 
of its poisonous property—a change corresponding to that 
which tetanine undergoes (p. 267). The gold salt is better 
adapted for preservation. The ordinary alkaloidal reagents 
produce in its solutions, if at all, only oily precipitates. 
As stated under mydatoxine, the formula of the hydro- 
chloride, C6II15N02.IIC1, is applicable to either one of two 
bases, C6H16NG2.OfI or C6II15N02. The base correspond- 
ing to the first formula is evidently a hoinologue of mus- 
carine, and should possess a similar physiological action. 
As a matter of fact, mytilotoxine does resemble muscarine 
somewhat in its action, and its occurrence together with 
betaine would seem to make it a decomposition-product of 
lecithin, in which case this base must be looked upon as a 
member of the choline group. It is interesting to know 
that a compound corresponding to the formula C6H16N02.0II 
has been known for some time, and was prepared by Har- 
riot in a manner analogous to Wurtz’s synthesis of chemistry of the ptomaines. 
257 
choline, by treating glycerin monochlorhydrine with tri- 
methylamine. This base, trimethyl-glyceryl-ammonium 
hydrate, has this structure: 
ch2oii 
I 
CHOU 
I 
ch2 
I 
N(CH3)3OH. 
It would seem that IIanriot’s base might possibly be 
identical with mytilotoxine, but a careful comparison made 
by Brieger showed that it possesses no physiological action 
and that its chemical reactions are entirely different. 
Mytilotoxine would, therefore, seem to possess the for- 
mula, C6H15N02, as originally given it by Brieger. From 
the fact that on distillation with potassium hydrate it yields 
trimethylamine, it follows that mytilotoxine is a quarter- 
nary base. He is inclined to regard it as a methyl deriva- 
tive of betaine, which is so common in mussels, and repre- 
sents it by formula No. 1. 
(1) 
co2h 
I 
CH.CH3 
I 
N(CH3)3.OH 
(2) 
ch2oii 
I 
C1I.CII3 
I 
N(CH3)3.OH 
No. 1, however, is C6H15N03, instead of C6H15N02, as 
above. The formula No. 2, C6H17N02, would represent 
a derivative of choline or muscarine, with only a slightly 
higher percentage of hydrogen. 
The Aurochloride, CcH15N02.HC1.AuC]3(Au = 41.6(3 
per cent.), crystallizes in cubes. Its melting-point is 182°. 
It is well to observe that Brieger has been unable to 
obtain this base from mussels that were allowed to putrefy 
for sixteen days. 
Physiological Action.—According to Brieger, mytilo- 
toxine produces all the characteristic effects seen in mussel 258 
BACTERIAL POISONS. 
poisoning, and it is, therefore, a strong paralysis-producing 
poison, and resembles curara in its action. This action is 
explainable now that Glause and Luchsinger have 
shown that all trimethyl-ammonium bases have a musca- 
rine-like action. For the symptoms induced by poisonous 
mussel see page 88. 
Gadinine, C7H17N02, was found in haddock (1885) 
which was allowed to decompose in open iron vessels for 
five days during summer. Briegeu has also obtained it 
from cultures of the bacteria of human feces on gelatin. 
The decomposing mass was thoroughly stirred every day 
in order to bring it into contact with atmospheric oxygen 
(Biiieger, I., 49). It was then treated with water, and 
hyd rochloric acid was added to acid reaction, and after being 
warmed the mixture was filtered and the filtrate concen- 
trated on the water-bath to a syrupy consistency. This 
syrupy residue was extracted with water, and the aqueous 
solution was precipitated with a solution of mercuric chlo- 
ride. The mercuric chloride precipitate contained a base, 
the quantity of which, however, was insufficient for a com- 
plete analysis (see page 272). The mercuric chloride filtrate, 
after the removal of the mercury by hydrogen sulphide, 
was evaporated to a syrup, and this was then repeatedly 
extracted with alcohol. The alcoholic solution thus ob- 
tained contained neuridine, a base of the same composition 
as ethylenediamine, muscarine, gadinine, and triethylamine. 
These bases were separated in the following manner: The 
alcoholic solution gave with platinum chloride a precipitate 
of neuridine. The filtrate from this platinum precipitate 
was heated on the water-bath to expel the alcohol, and then 
the platinum was removed by hydrogen sulphide. The 
aqueous filtrate was concentrated to a small volume which, 
on addition of platinum chloride, gave a precipitate of the 
isomer of ethylenediamine. The mother-liquor from this 
precipitate was concentrated on a water-bath, and on cool- 
ing the platinochloride of muscarine crystallized out. From 
the mother-liquor of this precipitate on standing in a des- 
iccator, the gadinine double salt crystallized. The mother- chemistry of the ptomaines. 
259 
liquor from the gadinine platinochloride was treated with 
hydrogen sulphide to remove the platinum, and the aque- 
ous filtrate on distillation with potassium hydrate gave 
triethylamine. 
Gadinine (from Gadus callarias, haddock) in small doses 
does not appear to be poisonous; larger doses (0.5-1 gramme) 
are decidedly toxic and may kill guinea-pigs. The formula 
of the free base as deduced from the analysis of the platiuo- 
chloride may be either C7H17N02 or C7H18N02 Oil. 
The Hydrochloride, C7H17N0,.IIC1, as obtained by 
the decomposition of the platinochloride with hydrogen 
sulphide, crystallizes under the desiccator in thick, colorless 
needles, which are easily soluble in water; insoluble in 
alcohol. It forms no combination with gold chloride, but 
does give crystalline precipitates with phosphomolybdic 
acid, phosphotungstic acid, and picric acid. 
The Platinochloride, (C7H17N02.HCl)2PtCl4 (Pt = 
27.68 per cent.), is at first quite soluble, and on standing over 
a desiccator it crystallizes in golden-yellow plates, which, 
when once formed, are again difficultly soluble in water. 
It can be recrystallized from hot water. It melts at 214°. 
Typhotoxine, C7H17N02.—This base was named thus 
by Brieger in 1885 (III., 86), and is regarded by him as 
the specific toxic product of the activity of Koch-Eberth’s 
typhoid bacillus. It is, however, probable that, as in the 
case of tetanus, there are basic and other products formed. 
He obtained it by cultivating the bacillus on beef-broth for 
eight to fourteen days at the temperature 37.5-38°. The 
nature of the soil on which it grows has a great deal to do 
with the formation of the poison. An especially important 
factor is the temperature : for Brieger has observed that 
no poison was produced in one case where the temperature 
remained by accident at 39° for twenty-four hours. In 
such cases creatine is present in quantity, whereas otherwise 
the reverse is the rule. 
In the process of extraction it occurs in the mercuric 
chloride precipitate, and from this it is obtained, after the 
removal of the mercury by hydrogen sulphide, as an easily 260 
BACTERIAL POISONS. 
deliquescent hydrochloride. This for the purpose of puri- 
fication is converted into the difficultly soluble aurochloride. 
Typhotoxine is isomeric with gadiniue and the compound 
C7H17NO,, which Brieger obtained from putrefying horse- 
flesh. In its properties it is, however, very different. 
Thus, the free base is strongly alkaline and its hydrochloride 
yields a difficultly soluble picrate. On the other hand, the 
isomer from horseflesh possesses a slightly acid reaction, 
and does not form a picrate. Again, typhotoxine gives 
with Ehrlich’s reagent (sulpho-diazobenzole) an imme- 
diate yellow color, which disappears upon the addition of 
alkali, whereas the isomer does not give this reaction. 
Furthermore, the two bases differ in their physiological 
action and in their behavior to alkaloidal reagents (see 
Table I.). Their aurochlorides, however, possess the same 
melting-point. 
The Hydrochloride is readily deliquescent, and unites 
with platinum chloride to form an easily soluble double 
salt crystallizing in needles. 
The Aurochloride, C7II17N02.IIC1.AuC13 (Au = 40.46 
per cent.), is difficultly soluble, and crystallizes in prisms, 
which melt at 176°. In its melting-point and solubility 
(197°, Brieger, Arch. f. pathol. Anat., 115, 489) it 
agrees with its isomer from horseflesh. From some of his 
first experiments in the cultivation of the typhoid bacillus, 
Brieger (II., 69) obtained a basic product differing in 
some of its characters from typhotoxine. Its aurochloride, 
on analysis, gave 41.91 and 41.97 per cent, of An, 16.06 
per cent, of C, and 3.66 per cent, of II.; while typho- 
toxine aurochloride gave 40.78 per cent. Au, 17.38 per 
cent. C, and 3.85 per cent. H. For a comparison of the 
reaction of these two substances see Table I. 
In its physiological action, typhotoxine differs from its 
isomer (page 262) in that the latter produces symptoms 
with well-marked convulsions, whilst the former throws 
the animal into more of a paralytic or lethargic condition. 
The action of this base has been studied only on mice and 
guinea-pigs. It produces at first slight salivation with 
increased respiration • the animals lose control over the 261 
CHEMISTRY OF THE PTOMAINES. 
muscles of the trunk and extremities, and fall down help- 
less upon their sides. The pupils become strongly dilated, 
and cease to react to light; the salivation becomes more 
profuse; the rate of heart-beat and of respiration gradually 
decreases, and death follows in from one to two days. 
Throughout the course of these symptoms the animals 
have frequent diarrhceic evacuations, but at no time are 
convulsions present. On post-mortem, the heart is found 
to be in systole, the lungs are strongly hyperaemic, the 
other internal organs pale, the intestines firmly contracted, 
and their walls pale. 
A Base(?), C7H17N02, was obtained by Brieger in 1880 
(III., 28) on working over about one hundred pounds of 
horseflesh which had been allowed to undergo slow putre- 
faction with limited access of air aud at a low temperature 
(—9° to +5°) for four months. It occurs in the mercuric 
chloride precipitate together with cadaverine, putrescine, 
and mydatoxine, and from these bases it can be separated 
and isolated according to the method on page 233. 
A similar, if not identical substance, having the com- 
position C7Hl7N02, was obtained by Bag insky and Stadt- 
iiagen (1890) from cultures on horseflesh, ten days at 35°, 
of a bacillus, closely allied to Finkler-Prior’s, and iso- 
lated from stools of cholera infantum. The gold salt in 
crystalline form and properties is the same as Brieger’s, 
except that it possesses a somewhat higher melting-point. 
The free substance possesses, even after most careful 
purification, a slightly acid reaction. This acidity is 
removed from even a large quantity of the substance by 
the addition of a drop of alkali. On account of the acid 
character of the free substance, Brieger does not consider 
it to be a base (a ptomaine). It differs, however, from the 
amido-acids in its poisonous character; in the fact that, 
unlike an acid, it does not unite with bases to form salts; 
and in not giving the characteristic red coloration (Hof- 
meister’s reaction for the amido-acids) with ferric chloride. 
Whatever the true nature of this substance may be, it 
nevertheless, in its other properties, behaves like a base. 262 
BACTERIAL TOISONS. 
Thus, it forms simple as well as double salts. On boiling 
with copper acetate, it gives amorphous floccules. Under 
the desiccator it solidifies into plates which deliquesce on 
exposure to the air. It does not combine either with silver 
oxide or with cupric hydrate. On dry distillation it yields 
a distillate possessing a strong acid reaction and a peculiar 
odor. The distillate does not give any precipitate with 
platinum chloride, or with gold chloride, nor does it react 
with copper acetate. With phosphomolybdic acid, how- 
ever, it forms an amorphous mass; with ferric chloride and 
potassium ferricyanide it yields an immediate precipitate of 
Berlin-blue, whereas the original substance does not give 
any blue coloration. 
The Hydrochloride, C7III7N02.HC1, crystallizes in 
fine needles which are insoluble in absolute alcohol. When 
its aqueous solution is treated with freshly precipitated 
silver oxide, the resulting filtrate contains some silver oxide 
in solution, from which it can be removed by hydrogen 
sulphide; thus differing from an ammoniacal silver solu- 
tion, which gives no precipitate on treatment with hydrogen 
sulphide. In this respect it resembles Salkowski’s base, 
page 231. For reactions of the hydrochloride, see Table I. 
The Aurochloride, C7H17N02.IIC1.AuC13, forms 
plates which are difficultly soluble in water, and melt at 
176°—the melting point of the gold salt of typhotoxine. 
It is dimorphous, since sometimes it is also obtained in 
needles which can be changed into plates. 
It does not form a pierate, nor does it give a reaction 
with sulpho-diazobenzole. 
Physiological Action.—This substance, when injected 
into frogs, produces a eurara-like action. A few minutes 
after the injection the animal falls into a condition of 
paralysis, and, although it can still react toward reflexes, it 
cannot move from its place. At times fibrillary twitchiugs 
pass over the body. The pupils dilate, the heart-action 
becomes gradually weaker, and finally, after several hours 
the animal dies, with the heart in diastole. Doses of 0.05 to 
0.3 gramme of the hydrochloride, injected into guinea-pigs, 
produce in a short time a slight tremor, gradual increase in CHEMISTRY OF THE PTOMAINES. 
263 
respiration, and slight moistening of the lower lip. The 
pupils at first contract, then dilate ad maximum, and become 
reactionless. The temperature remains at first normal; 
chills of short duration follow in rapid succession. The 
animal squats on the ground with its snout pressing against 
the floor in exactly similar manner as is caused by the 
mussel poison. Violent clonic convulsions follow in con- 
tinually shorter intervals, and at the same time lachryma- 
tion and salivation become profuse, but not as excessive 
as in the case of the muscarine-like ptomaines. The tem- 
perature sinks with the decrease in the rate of respiration, 
the ears previously gorged become pale and cold, and the 
heart-action becomes irregular and less frequent than before. 
General paralysis sets in, but the head still moves upward 
and backward. External stimuli induce violent clonic 
convulsions, the animal repeats frequently choking move- 
ments, and at the same time yields large quantities of 
saliva; finally, it falls upon its side completely paralyzed, 
and dies. The heart stops in diastole, the intestines are 
pale and strongly contracted, and the bladder is empty and 
likewise contracted. 
Morriiuic Acid, C9H13N03, was obtained by Gautier 
and Mourgues (1888) from brown cod-liver oil, together 
with six bases, already described—namely, butylamiue, 
amylamine, hexylamine, dihydrolutidine, aselline and mor- 
rhuiue. These bases constitute about 0.2 per cent, of the 
oil. The discoverers regard them as true leucomames, 
dissolved from the hepatic cells by the oil. It is more 
probable, however, that these compounds are the products 
of initial decomposition, and for that reason they are de- 
scribed under the head of ptomaines. This compound is 
relatively abundant, and is basic as well as acid in charac- 
ter. It is resinous in appearance, and can be crystallized 
in flattened prisms, or large lance-shaped plates. When 
recently precipitated it is oleaginous, viscous, then gradually 
hardens. It possesses a disagreeable aromatic odor re- 
sembling that of the sea-weeds, upon which the fish feed. 
According to the discoverers its probable source is the 264 
BACTERIAL POISONS 
lecithin derived thus from these weeds. It is soluble in 
alcohol, and but slightly in ether. It reddens turmeric, 
decomposes carbonates and with acids forms salts which 
precipitate lead acetate and silver nitrate, but not copper 
acetate, even on warming. 
The hydrochloride is crystalline, and is partially disso- 
ciated by excess of water. The platinum salt is soluble, 
and crystallizes in very small cross-shaped prismatic 
needles. The gold salt is amorphous and is readily altered 
on heating. 
The properties of this compound show that it is of a 
pyridine nature, and inasmuch as it does not give a pre- 
cipitate with copper acetate, it would appear that the carb- 
oxyl is not directly united to the pyridine nucleus. This 
does not necessarily follow now that we know that some 
amido-acids exists which do not give a reaction with copper 
acetate (see page 231). Its pyridine nature is further- 
more shown on distillation with lime. An oily alkaline 
base is thus obtained which forms an iodomethylate, and 
this with potassium hydrate yields quite an intense red 
color, resembling lees (De Coninck’s reaction). On 
oxidation with permanganate of potassium it yields a mono- 
basic acid. According to Gautier and Mourgues the 
compound is probably identical with De Jungh’s gaduine, 
and they ascribed to it the following constitution, which, 
it should be said, lacks full confirmation: 
II 
C 
\ 
HC COH 
I II 
H2C C—C3H6.C02H. 
\ / 
N 
H 
Compare with ty rosin, C9HuN03 (page 197). chemistry of the ptomaines. 
265 
A Base, C5H12N204, was obtained by Pouciiet (1884) 
from the residual liquors resulting from an industrial treat- 
ment of debris of bones, flesh, and waste of all kinds, with 
dilute sulphuric acid. It is accompanied by another base, 
C7H18N206, from which it can be separated by treatment 
with alcohol. The base itself forms tufts of delicate needles 
which alter or decompose less easily than the accompanying 
base. The platinochloride, (C5Hl2N2O4.H01)2PtCl4, forms 
a dull yellow powder, somewhat soluble in strong alcohol, 
but insoluble in ether. The platinochloride (C7II18N2()6. 
HCl)2PtCl4 is insoluble in ether. 
The hydrochlorides of these bases form silky needles 
which are altered by excess of hydrochloric acid and by 
exposure to air. Pouchet considers them to be closely 
allied to the oxy-betaines. The general alkaloidal reagents 
precipitate these bases; the phosphomolybdic precipitate, 
on addition of ammonia, gives a blue tint. Both bases are 
toxic, and exert a paralyzing action upon the reflex move- 
ments. 
The method employed by Pouchet for their isolation 
was to precipitate them as tannates. The precipitate was 
decomposed by lead hydrate in the presence of strong 
alcohol, the excess of lead removed from the solution by 
hydrogen sulphide, and the clear liquid thus obtained was 
submitted to dialysis. The above bases occurred in the 
dialysate. In the non-dialyzable portion volatile bases 
were found probably identical with those described by 
Gautier and Etard. 
Tetanine, C13H30N2O4, was obtained in 1886 by 
Brieger (III., 94) by cultivating impure tetanus microbes 
of Rosenbach, in an atmosphere of hydrogen on beef- 
broth for eight days at 37°—38°. It likewise occurs in 
cultures on brain-broth. Later (April, 1888), Brieger 
succeeded in obtaining tetanine from the amputated arm 
of a tetanus patient, identical in its physiological action 
and chemical reactions with that isolated from cultures 
of Rosenbach’s germs on beef-broth. The presence of 
tetanine during life in tetanus patients has thus been 266 
BACTERIAL POISONS. 
demonstrated. It lias not been found in the brain and 
nerve tissue of persons dead from tetanus. A portion of 
the jelly-like mass taken from the amputated arm was 
found to contain tetanus bacilli as well as staphylococci 
and streptococci, and when planted on beef-broth, tetanine 
was formed, but no tetanotoxiue or spasmotoxiue. 
Kitasato and Weyl (1890), employing pure cultures 
of the tetanus bacillus, obtained from kilogramme beef 
used as culture medium 1.7118 gramme of tetanine hydro- 
chloride (0.187 per cent.). Tetanotoxine was also present. 
For its isolation Brieger employed the following 
method: The cultures were slightly acidulated with 
hydrochloric acid, heated and filtered; the filtrate was 
then treated with lead acetate and with alcoholic mercuric 
chloride in the manner described under my.tilotoxine (page 
255). Kitasato and Weyl digest the cultures with 0.25 
per cent, hydrochloric acid for some hours at 60°, then render 
slightly alkaline, filter, and distil in vacuo at 60°. The 
residue in the retort is worked for tetanine by Brieger’s 
method, while the distillate contains tetanotoxine, ammo- 
nia, indol, hydrogen sulphide, phenol and butyric acid. 
The filtrate from the above mercuric chloride precipitate 
contains the greater part of the active principle, provided 
the precipitate has been thoroughly washed. After the 
removal of the mercury by hydrogen sulphide, it is evap- 
orated and the residue is repeatedly extracted with absolute 
alcohol, in which the tetanus poison readily dissolves, and 
can thus be separated from the insoluble ammonium 
chloride. The alcoholic solution is treated with alcoholic 
platinum chloride, which precipitates the ammonium and 
creatinine platiuochlorides, whilst the platinochloride of the 
poison remains in solution. The filtrate from this precipi- 
tate gives, on the addition of ether, a flocculeut precipitate 
possessing exceedingly deliquescent properties. The pre- 
cipitate is, therefore, rapidly filtered off by means of a 
pump, and dried in vacuo. It can then be recrystallized 
from hot 96 per cent, alcohol, and the beautiful clear-yellow 
plates thus obtained, if dried again in vacuo, become 
rather difficultly soluble in water, from which it can then CHEMISTRY OF THE PTOMAINES. 
267 
be recrystallized and obtained in a perfectly pure condi- 
tion. If boiled with boneblack it decomposes, yielding a 
non-poisonous crystalline compound. 
Phosphomolybdic acid cannot be used in the separation 
of tetanine, inasmuch as it destroys the poison (Brieger). 
Bocklisch has also observed that it destroys the poison 
formed in the putrefaction of fish. 
Tetanine obtained by treating the hydrochloride with 
freshly precipitated moist silver oxide forms a strongly 
alkaliue yellow syrup. With alkaloidal reagents it gives 
the same reactions as the hydrochloride, except that it does 
not give a blue color with ferric chloride and potassium 
ferricyanide. It is easily decomposed in acid solution, but 
is permanent in alkaline solution. 
The Hydrochloride, C]3H30N2O4.2HC], is deliques- 
cent and is easily soluble in absolute alcohol. Beside with 
platinum it combines only with phosphomolybdic acid to 
form an easily soluble crystalline precipitate, which on the 
addition of ammonium hydrate becomes white. If, how- 
ever, the hydrochloride is impure, phosphomolybdic acid 
produces a precipitate which is colored an intense blue by 
ammonia. Potassium-bismuth iodide yields. a precipitate 
which is at first amorphous, but soon becomes crystalline. 
Ferric chloride and potassium ferricyanidc produce a slowly 
developing blue color which probably is due to impurities. 
When kept for some months the highly poisonous hydro- 
chloride becomes syrupy, brownish, and wholly inert. 
Examined at this stage, the syrup was found, by means of 
platinum chloride, to contain a substance the hydrochlo- 
ride of which crystallized in plates. This is readily soluble 
in water and alcohol, and melts at 197°, with total decom- 
position, the same as tetanine. It combines only with phos- 
phomolybdic acid to form an easily soluble compound. The 
platinum salt has the composition C6II13N02 2HCl.PtCl4. 
This substance is non-poisonous and probably an amido- 
acid. It is different, however, from leuein and Nencki’s 
isomers of leuein, although possessing the same composi- 
tion. It is also isomeric with mydatoxine, C6H13N()2, but 
this is highly poisonous to mice, while the former is inert 268 
13ACTEKIAL POISONS. 
(see page 255). Tetanine resembles mytilotoxine with 
respect to this loss of toxicity on standing. 
The Platinochloride, C13H30N2O4.2HCl.PtCl4 (Pt = 
28.33 per cent.), is easily soluble in absolute alcohol from 
which it is precipitated on the addition of ether. From 
ninety-six per cent, alcohol it crystallizes in clear yellow 
plates. After repeated recrystallization from alcohol and 
drying in vacuo it becomes difficultly soluble in water so 
that it can be recrystallized from the latter. It decomposes 
at 197°. 
This base produces the characteristic, though probably 
not all the symptoms of tetanus, since we know of at least 
three other toxines (pages 194, 195) which occur with teta- 
nine in cultures of the tetanus microbe. The symptoms 
induced by relatively large doses in warm-blooded animals, 
as mice, guinea-pigs, and rabbits, exhibit two distinct phases. 
In the first, the animal is thrown into a lethargic paralytic 
condition, then suddenly becomes uneasy, and the respira- 
tion becomes more frequent. This is followed by the second 
phase, in which tonic and clonic convulsions, especially the 
former, predominate till death results. 0.5 gramme has 
but slight action on guinea-pigs. Small doses do not 
seem to affect guinea-pigs, while frogs seem to be much 
less sensitive than mice. The characteristic convulsions 
and opisthotonus seen in tetanus in man are also produced 
in guinea-pigs on injection of large doses of this base. Dogs 
and horses seem to be but slightly sensitive to the action of 
this poison. 
A Base, C14N20N2O4, was isolated by Guaresciii in 
1887 from putrid fibrin. It occurs in the chloroform or 
ether extracts along with the base C10H13N, and is probably 
an amido-acid (see page 201). 
A Base, C7H18N206, w7as isolated by Pouchet in 1884. 
It is said to form short, thick prisms which become brown 
when exposed to light. 
The Platinochloride, (C7H18N206.HCl)2PtCl4, crys- 
tallizes in prismatic needles which are insoluble in strong chemistry of the ptomaines. 
269 
alcohol. For further details in regard to this base see 
page 265. 
Tykotoxicon has been obtained in poisonous cheese 
(Vaughan, Wallace, Wolff), in poisonous ice-cream 
(Vaughan, Novy, Schearer, Ladd), in poisonous 
milk (Vaughan, Novy, Newton, Wallace, Firth, 
Schearer), and in cream-puffs (Stanton). The methods 
of separating this poison and its effect upon animals have 
already been given with sufficient detail. Chemically, it is 
very instable. When warmed with water to about 90°, it 
decomposes. Hydrogen sulphide also decomposes it, there- 
fore all attempts to isolate it by precipitation with some 
base, such as mercury or lead, and then removing the base 
with hydrogen sulphide, have failed. Its unstable char- 
acter is illustrated by the fact that it may disappear 
altogether within twenty-four hours from milk rich in the 
poison which is allowed to stand in an open beaker. 
With potassium hydrate it forms a compound which 
agrees in crystalline form, chemical reactions, and the per 
cent, of potassium which it contains, with the compound of 
diazobenzole and potassium hydrate. This substance is best 
obtained from milk containing tyrotoxicon as follows : The 
filtered milk, which is acid in reaction, is neutralized with 
sodium carbonate, agitated with an equal volume of ether, 
allowed to stand in a stoppered glass cylinder for twenty- 
four hours, the ether removed, and allowed to evaporate 
spontaneously from an open dish. The aqueous residue is 
acidified with nitric acid, then treated with an equal volume 
of a saturated solution of potassium hydrate, and the whole 
concentrated on the water-bath (this compound is not 
decomposed below 130°). On being heated the mixture 
becomes yellowish-brown, and emits a peculiar aromatic 
odor. On cooling the tyrotoxicon compound forms in 
beautiful, six-sided plates along with the prisms of potas- 
sium nitrate. 
With equal parts of sulphuric and carbolic acids, pure 
tyrotoxicon gives a green coloration, but in whey the color 
varies from yellow to orauge-red. This color reaction may 270 
BACTERIAL POISONS. 
be used as a preliminary test in examining milk for tyro- 
toxicon. It is best carried out as follows: Place on a clean 
porcelain surface two or three drops each of pure carbolic 
and sulphuric acids. Then add a few drops of the aqueous 
solution of the residue left after the spontaneous evapora- 
tion of the ether. If tyrotoxicon be present, a yellow to 
orange-red coloration will be produced. This test is to 
be regarded only as a preliminary one, for the coloration 
may be due to the presence of a nitrate or nitrite, or as 
Huston has shown, to butyric acid. The tyrotoxicon 
must be converted into the potassium compound and puri- 
fied before the absence of nitrate or nitrite can be positively 
demonstrated. Moreover, the physiological test should 
always be made in testing for this poison. 
With platinum chloride in alcoholic solution tyrotoxicon 
forms a compound which explodes with great violence at 
the temperature of the water-bath. This also corresponds 
with the compound of platinum chloride and diazobenzole. 
Pure tyrotoxicon is insoluble in ether, and its extraction 
from alkaline solutions by this solvent is due to the pres- 
ence of foreign matter, with which the poison is taken up 
by the ether. 
The physiological action of this ptomaine has been suf- 
ficiently discussed in a preceding chapter. 
Mydaleine (/ivdaMor, putrid) is a poisonous base ob- 
tained in 1885 from putrefying cadaveric organs, liver, 
spleen, etc. (Brieger, IT., 31, 48). Though it is appa- 
rently present on about the seventh day, it is unobtainable 
until about the third or fourth week. The method for its 
separation from the accompanying bases is given under 
Sapriue (page 220). It is liable to occur in the mercuric 
chloride filtrate, as well as in the precipitate, inasmuch as 
the double salt is insoluble only in perfectly absolute alco- 
hol. In order to purify the platinochloride obtained as on 
page 221, it is repeatedly recrystallized from a very small 
quantity of lukewarm water. This base has not been ob- 
tained in sufficient quantity to permit of a complete deter- 
mination of its composition. It is probably a diamine, CHEMISTRY OF THE PTOMAINES. 
271 
containing four or five carbon atoms, and hence is nearly 
related to some of the diamines already described. 
The Platinochloride, on analysis, gave : Pt = 38.74 
C = 10.83, H = 3.23. It crystallizes in small needles, and 
is extremely soluble in water. 
The Hydrochloride crystallizes with extreme dif- 
ficulty, even on standing for some time in a desiccator. On 
exposure to the air it rapidly deliquesces. 
Physiological Action.—Mydaleine has an entirely specific 
action. Small quantities injected into guinea-pigs or 
rabbits produce, after a short time, a moistening of the 
under lip, and an abundant flow of secretion from the nose 
and eyes. The pupils dilate gradually to maximum, and 
become reactionless; the ear vessels become strongly in- 
jected, and the body temperature rises 1° to 2°. The hairs 
bristle, and the animal occasionally shudders. Gradually 
the salivation ceases,the respiration and heart-action, which 
were at first hastened, now decrease, the temperature falls, 
the ears become pale, and the animal finally recovers. 
During the action of the poison the animal shows a ten- 
dency to sleep, and the peristaltic action of the intestines is 
heightened. Larger doses (0.050 gramme) induce au ex- 
ceedingly violent action, which invariably results in the 
death of the animal. On post-mortem, the heart is found 
to be stopped in diastole, and the intestines and bladder 
contracted; otherwise nothing abnormal is observed. 
A Toxic Base.—From human livers and spleens which 
were decomposing for two weeks in thorough contact with 
air there was isolated, besides cadaverine and putrescine, a 
small quantity of a poisonous base (Brieger, II., 29, 48). 
The mercuric chloride precipitate was decomposed, and the 
hydrochlorides were precipitated by gold chloride (to re- 
move cadaverine, which is soluble), and the auroehloride 
was then changed into the platinum salt, whereby the in- 
soluble putrescine platinochloride was removed. In the 
mother-liquors from the putrescine salt an easily soluble 
platinum compound was separated, and found to contain 
41.30 per cent. Pt. It crystallized in tine needles. The 272 
BACTERIAL POISONS. 
hydrochloride formed small, readily deliquescent needles, 
and did not produce a precipitate in alcoholic platinum 
chloride. Injected into guinea-pigs and rabbits it induced 
an exalted peristaltic action of the intestines, which lasted 
several days, and produced in the animals, on account of 
the continuous evacuations, a condition of great weakness. 
No disturbance in the functions of the other organs was 
observed. 
A Base was isolated from decomposing haddock which 
were exposed for five days during summer in an open iron 
vessel. Brieger (I., 42) found in the aqueous mercuric 
chloride precipitate (see page 258) a base the hydrochloride 
of which crystallized in well-formed, small needles. The 
platinochloride likewise crystallized in beautiful needles, 
and gave, on analysis, 36.03 per cent, of Pt; 7.81 per cent, 
of N. 
A substance of muscarine-like action was obtained by 
Brieger (I., 59) from putrefying gelatin, ten days at 
35°, though in insufficient quantity to permit a determina- 
tion of its character. The residue containing this substance 
gave, on distillation with alkali, only ammonia. 
A Base was obtained by Bockliscii (III., 52, 53) from 
herring which had undergone putrefaction for twelve days. 
It was found in the distillate, together with trimethylamine 
and dimethylamine, obtained by distilling the mercuric 
chloride filtrate, after the removal of the mercury, with 
sodium hydrate. The platinochloride was easily soluble, 
and crystallized in large thin plates. On analysis it gave: 
Pt = 28.57, C = 22.34, II — 4.66. The hydrochloride is 
easily soluble in water, and in absolute alcohol, and be- 
sides with platinum gives only with phosphomolybdic acid 
a yellow precipitate which is soluble in excess, and with 
ammonia gives an immediate blue color. It immediately 
reduces a mixture of ferric chloride and potassium ferri- 
cyanide with formation of Berlin blue; and similarly CHEMISTRY of the ptomaines 
273 
throws down metallic gold from solutions of gold 
chloride. 
From poisonous mussel, Brieger (III., 79) obtained an 
aurochloride of a base crystallizing in needles. The quan- 
tity isolated was insufficient for analysis. It is interesting 
because of its property of inducing salivation, a symptom 
which has been observed by Sohmidtmann and by Crumpe 
in some cases of mussel poisoning. 
A Base was obtained by Guareschi and Mosso (Journ. 
fur praktische Chew., 28, 508) from fresh beef, in the 
alkaline ether extract obtained by Dragendorff’s method. 
It formed a yellowish alkaline fluid, of unpleasant odor, 
and after a time gave a deposit of microscopic crystals. 
The hydrochloride gave the following reactions : Gold chlo- 
ride, yellow crystalline precipitate ; platinum chloride, pre- 
cipitate ; potassium iodide and iodine in hydriodic acid, 
kermes-red precipitate; phosphotungstic acid, nothing; 
phosphomolybdic acid, an abundant yellow precipitate; tan- 
nic acid, heavy, grayish precipitate, same with Mayer’s 
reagent; picric acid, yellow precipitate; MarmiVs reagent, 
precipitate soluble in excess ; potassium bichromate, noth- 
ing ; potassium permanganate and sulphuric acid, violet 
color ; potassium ferricyauide and ferric chloride, Prussian 
blue precipitate. 
By giving a precipitate with tannin, and not with phos- 
photungstic acid, it resembles neurine. 
Ch. Gram has studied the decomposition of yeast under 
the influence of an infusion of hay. The yeast was allowed 
to putrefy for fourteen days, and was then treated with zinc 
sulphate. The latter was precipitated by barium hydrate, 
and the filtrate after the removal of the barium by sul- 
phuric acid, was evaporated to dryness, and extracted with 
absolute alcohol. The alcoholic solution was evaporated, 
and the residue again extracted with alcohol. The extrac- 
tion residue was taken up with water, and again subjected to 
the above treatment with zinc sulphate, barium hydrate, etc. 274 
BACTERIAL POISONS. 
The filtrate was poisonous, and produced, in frogs, paral- 
ysis and stoppage of the heart in diastole Addition of 
platinum chloride and alcohol precipitated two bases. One 
of these, although possessing a curara-like action, did not 
affect the heart. When its solution was heated for twenty- 
four hours on the water-bath, it caused general paralysis 
and stoppage of the heart. The platinochloride contained 
38.05 per cent, of platinum. 
The other base also possessed a slight curara-like action, 
and its platinochloride gave, on analysis, 40.92 and 39.4 
per cent, of platinum. 
Brieger found a basic substance in small quantities in 
cultures of the staphylococcus pyogenes aureus on bouillon 
and beef-broth (II., 74). The hydrochloride formed groups 
of colorless, non-deliqucscent needles. With platinum 
chloride it yielded a double salt, crystallizing in needles, 
and containing 32.93 per cent, of Pt. For its reactions, 
see Table I. 
From aqueous as well as alcoholic solutions of cultures of 
staphylococcus aureus Leber (1888) isolated a crystalline 
substance which he named phlogasine. The composition 
of this substance is not known. It does not seem to con- 
tain nitrogen, and inasmuch as it blackens silver it prob- 
ably contains sulphur. It crystallizes in fine needles which 
are soluble in ether and in alcohol; difficultly soluble in 
water. It sublimes in needles. Alkalies precipitate it as 
amorphous yellow fioccules which arc soluble in acid and 
then can be recrystallized. With potassium ferricyanide 
and ferric chloride it yields a blue color, and with potassio- 
mercuric, cadmic, and bismuth iodides precipitates which 
are soluble in excess. No precipitate is produced by gold 
or platinum chlorides, phosphotungstic or molybdic, tan- 
nic or picric acids. 
A small quantity applied to the conjunctiva produces 
intense inflammation, suppuration, and necrosis. Intro- 
duced into the anterior chamber it induces intense suppura- 
tion and keratitis. The substance is entirely distinct from 
the base obtained by Brieger, described above. CHEMISTRY OF THE PTOMAINES. 
275 
A Base—boiling point about 284°—was obtained by 
Brieger (II., Gl) from human livers and spleens which 
were putrefying for two to three weeks. It occurs in the 
mercuric chloride filtrate, as described under Saprine, page 
220, together with some mydaleine, trimethylamine, and 
hydrocarbons. The filtrate, after the mercury is removed 
by hydrogen sulphide, is evaporated to dryness, and finally 
the last traces of water are removed in a vacuum. The 
residue is then treated with absolute alcohol, and from this 
alcoholic solution the mydaleine is precipitated by the addi- 
tion of alcoholic mercuric chloride. The trimethylamine 
is separated by distillation of the alkaline filtrate, previously 
deprived of its mercury by hydrogeu sulphide; while the 
mother-liquor yields an oily mixture of hydrocarbons and 
bases. The latter were separated by fractional distillation, 
whereby only one of the bases was obtained in sufficient 
quantity for study. It boiled at about 284°, and gave 
with hydrochloric acid, oil evaporation, a salt crystallizing 
in beautiful, long needles, which were very easily soluble 
in perfectly absolute alcohol. With gold chloride and picric 
acid it gave only oily products; with ferric chloride and 
potassium ferricyanide, an intense blue; with platinum 
chloride, an extremely easily soluble double salt, which 
appeared under the microscope in the form of very fine 
needles, while from alcohol-ether the double salt slowly 
separated in thin plates which contained 30.36 per cent, of 
platinum. The free base showed a slight fluorescence. It 
is not poisonous, and, according to Brieger, is probably 
a pyridine derivative. 
Other non-poisonous bases were present in very small 
quantity in the mother-liquor described above, after the 
separation of the oily mixture. 
Peptotoxine.—By this name Brieger (I., 14-19) has 
designated a poisonous base which he has found in some 
peptones, and hence in the digestion of fibrin ; in putre- 
fying albuminous substances, such as fibrin, casein, brain, 
liver, and muscles. It is a well-known fact that animal 
tissues, in the early stages of putrefaction, possess strong 
toxic properties, even before the decomposition could have 276 
BACTERIAL POISONS. 
advanced far enough to etlect a splitting-up of the proteid 
and carbohydrate molecules. Brieger and others have 
tried to seek an explanation of this toxicity by connecting 
it with an early peptonization of the proteids brought about 
by the action of ferments which are distributed throughout 
the tissues, and which begin their activity immediately after 
death. This poison has not been definitely isolated, but its 
general properties and action have been studied by 
Brieger and Salkowski. The former prepared it by 
digesting fibrin for twenty-four hours with gastric juice at 
the temperature of the blood. The perfectly fresh peptone 
thus obtained was evaporated to a syrupy residue, and this 
was then extracted with boiling alcohol. The residue left 
on evaporation of the alcoholic solution was digested for 
some time with amyl alcohol, which on subsequent evapor- 
ation gave amorphous brownish masses. This extract can 
then be purified by neutral lead acetate. The filtrate, after 
the removal of the lead by hydrogen sulphide, is repeat- 
edly extracted with ether, then evaporated to dryness, and 
extracted as before, with amyl alcohol. This final extract 
is evaporated to drive off* the alcohol, taken up with water, 
and filtered. The colorless aqueous solution thus obtained 
contains the poisonous substance, which, however, can only 
with extreme difficulty be brought to crystallization in 
vacuo. 
This poison, when in its purest condition, as shown by 
its failure to give the biuret reaction, possesses a neutral 
reaction. Its behavior to Millon’s reagent is quite charac- 
teristic: it gives a white precipitate, which on boiling 
becomes intensely red. From this reaction, Brieger is 
inclined to regard this substance as a hydroxyl or an 
amido-derivative of benzole. The ptomaine can be ex- 
tracted from acid as well as alkaline solution by amyl 
alcohol—more difficult in the cold than on heating. It 
is absolutely insoluble in ether, benzol, and chloroform ; 
very soluble in water. It is not destroyed by boiling, by 
passing hydrogen sulphide, or by strong alkalies; but is 
destroyed, however, when the putrefaction lasts longer 
than eight days. For its behavior to reagents, see 
Table I. CHEMISTRY OF THE PTOMAINES. 
277 
Various observers have shown that peptone possesses a 
toxic action, and some have been led to regard this toxicity 
as not due to the peptone itself, but rather to the presence 
of this or some other ptomaine. At least Brieger found 
one specimen of dry Witte’s peptone to be perfectly harm- 
less; whereas, the fresh peptone formed by fibrin digestion 
possessed strong toxic powers. Moreover, this non- 
poisonous peptone when exposed to the action of gastric 
juice was found to yield the poisonous substance. The 
poisonous nature of proteids aud the physiological action 
of this base will be described later. 
Pyocyanine, C14H14N02, is the coloring matter of blue 
pus, and is produced by the action of bacillus pyocyaneus. 
It was isolated by Ledderhose (1887) and is said to be an 
anthracene derivative. On contact with the air it is oxidized 
to pyoxanthose, a yellow substance. According to Kunz 
it contains nitrogen and sulphur. The picrate is of a dark 
reddish-brown color; the platinum salt is black and some- 
times is obtained as glittering fine golden needles. 278 
BACTERIAL POISONS. 
Table of Ptomaines. 
Formula. 
Name. 
Discoverer. 
Physiological action.1 
C H6N 
Methylamine. 
Bocklisch. 
Non-poisonous. 
C2 Hj N 
Dimethylamine. 
Brieger. 
tt tt 
C3H9N 
Trimethylamine. 
Dessaignes. 
tt (( 
c2h5n 
Spermine(?). 
Kunz. 
ft tt 
c2 h7 n 
Ethylamine. 
Hesse. 
it ft 
C4 HUN 
Diethylamine. 
Bocklisch. 
it -ft 
c6 H1BN 
Triethylamine. 
Brieger. 
it tt 
O3 -N 
Propylamine. 
“ 
C4 H„N 
Butylamine. 
Gautier & Mourgues. 
Poisonous(?). 
C6 HUN (?) 
Tetanotoxine. 
Brieger. 
Poisonous. 
c6 h13n 
Amylamine. 
Hesse. 
“ 
c6 h1bn 
Hexylamine. 
“ 
“ 
O7 HUK 
Di-hydrolutidine. 
Gautier & Mourgues. 
“ 
C8 HUN 
Collidine(?). 
Nencki. 
C8 HUN 
Pyridine base(?). 
0. de Coninck. 
c8 h18n 
Hydrocollidine(?). 
Gautier and Etard. 
Poisonous. 
C9 H13N 
Parvoline(?). 
“ “ “ 
Ci0H,6N 
Unnamed. 
Guareschiand Mosso. 
Poisonous. 
Pyridine base(?). 
0. de Coninck. 
OioUijN 
Hydrocoridine(?). 
Griffiths. 
'-taHglJN 
Unnamed. 
Delezinier. 
C2 H8 No 
Ethylidenediamine(?). 
Brieger. 
Poisonous. 
C3 Hg N2 
Trimethylenediamine(?). 
tt 
«« 
C4 h12n2 
Putrescine. 
tt 
Not very poisonous. 
C5 H14N2 
Cadaverine. 
t t 
tt tt 
GB H14N2 
Neuridine 
Non-poisonous. 
c6 h14n2 
Saprine. 
“ <« 
c7 ii10n2 
Unnamed. 
Morin. 
tt a 
o10h26n2(?) 
Susotoxine. 
Novy. 
Poisonous. 
c,h7 n3 
Methyl-guanidine. 
Brieger. 
“ 
C19H07M3 
Morrhuine. 
Gautier & Mourgues. 
Diuretic, etc. 
Oi3H20N4 
Unnamed. 
Oser. 
Ol7®38^4 
“ 
Gautier and Etard. 
Aselline. 
Gautier & Mourgues. 
Poisonous. 
c6 H13N 0 
Neurine. 
Brieger. 
it 
Cg HUN 0 
My dine. 
“ 
Non-poisonous. 
CB HjjN 02 
fc-amido-valerianic acid. 
E. and H. Salkowski. 
(( “ 
c6 h15n o2 
Choline. 
Brieger. 
Poisonous. 
c0 h13n o2 
Mydatoxine. 
C6 H]gN 02 
Unnamed. 
Brieger, 1888. 
(tetanus cult.) 
Non-poisonous. 
C6 h1bn 02 
Mytilotoxine. 
Brieger. 
Poisonous. 
1 Only those bases are here denoted as poisonous which possess a decided toxicity. CHEMISTRY OF THE PTOMAINES. 
279 
Table op Ptomaines—Continued. 
Formula. 
Name. 
Discoverer. 
Physiological action.1 
C7 H17N 02 
Gadinine. 
Brieger. 
Poisonous. 
C7 h17n 02 
Typhotoxine. 
“ 
“ 
C7 h17n 02 
Unnamed. 
“ 
Ci4H14N 02 
Pyocyanine. 
Ledderhose. 
Non-poisonous. 
C6 H1SN 03 
Betaine. 
Brieger. 
c5 h16n 03 
Muscarine. 
“ 
Poisonous. 
C9 H13N O3 
Morrhuic acid. 
Gautier & Mourgues. 
c5 h12n2o4 
Unnamed. 
Pouchet. 
Poisonous. 
Ci3H3qN204 
Tetanine. 
Brieger. 
“ 
c14h20n2o4 
Unnamed. 
Guareschi. 
c7 h18n2o6 
“ 
Pouchet. 
Poisonous. 
tt 
Tyrotoxicon. 
Vaughan. 
«( 
Mydaleine. 
Brieger. 
<< 
Spasmotoxine. 
U 
A diamine(?). 
“ (tetanus cult.) 
Peptotoxine. 
“ 
ft 
Phlogosine. 
Leber. 
Inflammatory. 
1 Only those bases are here denoted as poisonous which possess a decided toxicity. CHAPTER XII. 
CHEMISTRY OF THE LEUCOMAINES. 
Under this head are classed those basic substances which 
are found in the living tissues, either as the products of 
fermentative changes or of retrograde metamorphosis. 
Most of these substances have already been known for 
many years, though their real significance as alkaloidal 
bodies, and their relation to the functional activities of the 
animal organism have been but little understood, or rather 
they have not been brought together under the leading 
conception that they are alkaloidal products of physiologi- 
cal change. The first attempt at the systematic study and 
generalization of these basic substances was made by 
Gautier, who applied to them the name leucomaines, a 
term derived from the Greek signifying white of 
eggs. Under this name he includes all those basic sub- 
stances which are formed in animal tissues during normal 
life, in contradistinction to the ptomaines or basic products 
of putrefaction. The distinction between vegetable and 
animal alkaloids is not very well defined, and, in fact, there 
seem to be reasons for considering their formation as due 
to the same causes which bear an intimate relation to the 
physiology of the cells and tissues of both kingdoms. 
Thus, vegetable tissues are known to contain not only 
definite ptomaines, such as choline, but also leucomaines, as 
hypoxanthine, xanthine, etc. Indeed, in this latter group 
must be placed, on account of their relation to xanthine, 
those well-defined alkaloidal bases, caffeine and theobro- 
mine. Not only are the representatives of these two 
divisions of basic substances common to both kingdoms, 
but their parent bodies, lecithin, nuclein, etc., are known 
to occur in both, thus giving rise to the same bases on 
decomposition. CHEMISTRY OF THE LEUCOMAINES. 
281 
So far as the genesis of most of the leucoma'ines is 
concerned, we know very little, though Gautier is of the 
belief that they are being formed continuously and inces- 
santly in the animal tissues side by side with the forma- 
tion of urea and carbonic acid and at the expense of the 
nitrogenous elements. It is quite probable, as Kossel has 
pointed out, that some of these products are in themselves 
antecedents of end-products of metabolism. This is unques- 
tionably true of the imido group, which exists in the ade- 
nine and guanine molecules, and through vital or putre- 
factive processes is split off, giving rise to ammonia, which 
in turn serves to form urea and uric acid. Bouchard has 
sought an explanation of the presence of these bases in the 
urine, by supposing that they were originally formed in 
the intestinal tract, from which they were absorbed into the 
system, to be subsequently eliminated by the kidneys. This 
view has also been brought forward by Schar (1886), who 
holds that these bases, which may be formed by putrefactive 
changes in the intestinal tract, are absorbed into the circu- 
latory system, whence they may be partly eliminated by 
the kidneys or may be partly deposited in the tissues them- 
selves. 
The views of Bouchard and Schar have, to a certain 
extent, been confirmed by the investigations of Udranszky 
and Baumann, who showed that the well-known ptomaines, 
cadaverine and putrescine, occur in the urine in cystinuria, 
and are formed by putrefactive changes induced in the in- 
testinal tract probably by specific microorganisms. Under 
this same head fall the recent observations of Wolkow 
and Baumann, that alkapton is produced from tyrosin by 
similar changes in the intestines. The origin of the true 
leucoma’ines cannot, however, be accounted for in this 
manner, for they are indissolubly connected with the meta- 
bolism of the cell itself, and are, therefore, formed in the 
tissues and organs proper, especially those rich in nucleated 
cells. 
Another source of the nitrogenous bases must not be 
lost sight of, and that is protoplasm itself. The researches 
of Drechsel, Siegfried, and Schulze have shown that 282 
BACTERIAL POISONS. 
nitrogenous bases do result from the decomposition of 
animal and vegetable proteids (see p. 242). 
The leucoma'ines proper can be divided into two distinct 
and well-defined groups: (1) the Uric Acid Group, and 
(2) the Creatinine Group. 
The first of these divisions contains a number of well- 
known bases which are closely related to uric acid. The 
order in which they will be described is as follows : 
Adenine, C5H5N6. 
Hypoxanthine, C5H4N40. 
Guanine, C6H5NaO. 
Xanthine, C5H4N402. 
(Uric Acid, C5H4N403.) 
Heteroxantliine, C6H6X402. 
Paraxanthine, C7H8N402. 
Carnine, C7H8N403. 
Pseudoxanthine, C4H5N40. 
Gerontine, C5H14X2. 
Spermine, C2H5N ( ? ). 
The members of the second group have all been dis- 
covered by Gautier, and by him are regarded as allied 
to creatine and creatinine. These two substances, especially 
the latter, have been hitherto regarded as strongly basic in 
character, but Salkowski has recently shown that creati- 
nine, when perfectly pure, possesses little or no alkaline 
reaction, and, moreover, does not combine with acids. The 
bases in this group are : 
(Creatinine, C4H7N30.) 
(Creatine, C4H9N302.) 
Cr uso - creati n i n e, C5H8N40. 
Xantho-creatinine, C5H10N4O. 
Amphi-creatine, C9HlfN704. 
Base, CnH24N10O6. 
Base, C12H25NuO, 
Besides these two general classes of leucoma'ines, there 
may be made a third class of undetermined leueomames, 283 
chemistry of the leucomaines. 
embracing those bases which have been observed, but 
studied more or less incompletely, in the various physio- 
logical secretions of the body. 
Leucomaines of the Uric Acid Group. 
Adenine, C5H5H5, which was discovered by Kossel in 
1885, forms the simplest member of the uric acid group of 
leucomaines, and as such it deserves special attention, inas- 
much as it shows most clearly the relation that exists 
between hydrocyanic acid and the members of this group. 
This base is apparently formed by the polymerization of 
hydrocyanic acid—a view that is confirmed, at least in 
part, by the fact that on heating with potassium hydrate 
to 200°, it yields a large quantity of potassium cyanide. 
Moreover, by the action of reducing agents, it is converted 
into a substance similar to, if not identical with, azulmic 
acid. It has not been prepared synthetically, though 
Gautier has claimed to have synthesized two closely re- 
lated bodies, xanthine and methyl-xantlnnc, by simple 
heating of hydrocyanic acid in a sealed tube in contact with 
water and a little acetic acid. 
This base was first prepared from pancreatic glands— 
lienee the term adenine, which is derived from the Greek 
word a6rjv, meaning a gland. It has since been shown to 
occur together with guanine, hypoxanthine, etc., as a 
decomposition-product of nuclein, and, therefore, it may 
be obtained from all tissues and organs, animal or vege- 
table, rich in nucleated cells. Accordingly, it has been 
found in the kidneys, spleen, pancreatic, thymus and lym- 
phatic glands, in beer-yeast, in spermatic fluids, but not in 
testicles of the steer ; occurs also in tea-leaves. In the latter 
adenine appears to exist in a preformed condition, since it can 
be extracted without the use of acid reagents. The thymus 
gland, as a prototype of embryonic, highly cellular tissue, 
yields a considerable amount of adenine ; that from a calf, 
for instance, was found by Schindler to contain 0.18 per 
cent. It has also been observed in the liver and urine of 284 
BACTERIAL POISONS. 
leucocythsemic patients ; its occurrence in this disease will 
be readily understood when it is remembered that leucocy- 
thsemia is characterized by the presence in the blood of an 
unusual proportion of the nucleated white blood-corpuscles, 
which, owing to various unfavorable conditions, become de- 
stroyed in time, and the contained nuclein, as a result, 
splits up into adenine and guanine. These two bases may, 
therefore, be expected in all pathological conditions where 
there is an abnormal accumulation of pus. Indeed, as early 
as 1865, Naunyn extracted from pus, obtained from the 
pleural cavity, a considerable quantity of a substance 
which was probably either adenine or guanine, or both. 
Adenine does not occur, or only in minute traces, in meat 
extract; and in this it resembles guanine, which is present 
only in traces. This may be due to the fact that adenine 
and guanine are readily converted into hypoxanthine and 
xanthine respectively, as has been shown in the putrefaction 
experiments of Schindler. They may be considered as 
transitional products of cell-metabolism, the imido group 
contained in each readily being replaced by oxygen, and 
giving rise to ammonia, and this in turn to urea. Kossel, 
however, explains this fact on the ground that the muscle 
tissue is very poor in nucleated cells, i. e., in nuclein. It 
would seem that the muscle cell in losing the morphological 
character of a cell has also suffered a corresponding loss in 
its chemical properties. For while the decomposition- 
products of nuclein—hypoxanthine, xanthine, phosphoric 
acid, etc.—are found in the muscle tissue, they do not exist 
in combination as they do in the nuclein molecules. This is 
seen in the fact that the bases exist in the free condition, 
since they can be extracted by water; and again, the phos- 
phoric acid is present in the muscle tissue, not in organic 
combination, but as a salt. In the nucleated cell, adenine, 
guanine, etc., do not exist in the free condition, but form, 
in part at least, with albumin and phosphoric acid, a loose 
combination which is readily decomposed by the action of 
acids at the boiling temperature. This same change takes 
place spontaneously after death. 
There can be no doubt that adenine and guanine play an CHEMISTRY of the leucomaines. 
285 
important part in the physiological function of the cell 
nucleus, which, from recent observations, appears to be 
necessary to the formation and building up of organic 
matter. It is now known that non-nucleated cells, though 
capable of living, are not capable of reproduction. We 
must look, therefore, to the nucleus as the seat of the 
functional activity of the cell—indeed, of the entire organ- 
ism. Nuclein, the parent substance of adenine and guanine, 
is the best known and probably most important constituent 
of the nucleus, and as such it has been already credited 
with a direct relation to the reproductive powers of the 
cell. This chemical view has recently been confirmed by 
Zacharias, who showed that chromatin of histologists is 
identical with nuclein. Liebermann has questioned nu- 
clein as being the source of xanthine compounds, but in 
this he is not supported by the mass of evidence. 
The method employed by Kossel for the preparation 
of adenine, is as follows : The finely divided pancreatic 
glands are heated to boiling, for three or four hours, with a 
large quantity of dilute sulphuric acid (0.5 per cent, by 
volume of concentrated acid), and the acid solution thus 
obtained is treated with a slight excess of hot concentrated 
baryta water. The excess of baryta is removed by carbonic 
acid, and the solution is then filtered; the filtrate is con- 
centrated to a small bulk, about 100 c.c., rendered alkaline 
with ammonium hydrate, and finally precipitated with an 
ammoniacal solution of silver nitrate. The precipitate, 
consisting of the silver compound of the xanthine bodies, 
is partially dried by spreading over porous porcelain 
plates; then dissolved in warm nitric acid of specific 
gravity 1.1, to which a little urea has been added to pre- 
vent the formation of hypoxanthine should traces of nitrous 
acid be present. The filtered acid solution, treated with 
silver nitrate, on cooling, gives a deposit of the silver 
salts of hypoxanthine, guanine, and adenine, which is fil- 
tered off and thoroughly washed. The adenine separates 
out almost quantitatively if a little silver nitrate solution 
is added. The filtrate contains any xanthine silver com- 
pound that may be present. The washed precipitate of the 286 
BACTERIAL POISONS. 
silver salts is suspended in water, nitric acid added, decom- 
posed with hydrogen sulphide (ammonium sulphide, or, 
better, hydrochloric acid, may be used), and the clear filtrate 
is concentrated on the water-bath to a small volume. It is 
then saturated with ammonium hydrate and digested on the 
water-bath for some time, whereby adenine and hypoxan- 
thine go into solution, while the guanine remains undis- 
solved (see p. 287). From the annnoniacal solution on 
partial concentration and subsequent cooling, the adenine 
crystallizes out first, whereas the more soluble hypoxanthine 
remains in solution. If the adenine is still colored it can 
be purified by dissolving in water and boiling with animal 
charcoal. The hot aqueous solution is then rendered very 
slightly alkaline with ammonium hydrate and allowed to 
cool; adenine crystallizes out, and can be still further puri- 
fied by recrystallization from water. 
Ammonium sulphide has been employed by Schindler, 
in place of hydrogen sulphide, in decomposing the silver 
compounds of the above bases. Bruhns recommends in- 
stead warming with very dilute hydrochloric acid, espe- 
cially if guanine is present. The solution can then be 
neutralized with FalIC03, using methyl-orange as indi- 
cator, and the adenine separated from hypoxanthine by the 
picric acid method described below. 
Another method for the separation of adenine from 
hypoxanthine is based upon the behavior of the nitrates 
of these bases in aqueous solution. From concentrated 
aqueous solutions of the nitrates, free hypoxanthine crystal- 
lizes out first, because the nitrate is decomposed; whereas, 
adenine, which is a stronger base, remains in combination 
with the acid, in solution. 
Schindler determines adenine and hypoxanthine indi- 
rectly. The ammoniacal solution which is filtered from the 
insoluble guanine is evaporated to dryness on a weighed 
platinum dish, dried at 110°, and weighed. A nitrogen 
determination is now made of the mixed bases and from 
these data the proportion of each is calculated. 
By far the best method for the quantitative separation of 
adenine and hypoxanthine is the picrate method of Bruhns. CHEMISTRY OF THE LEUCOHAINES. 
287 
The solution of the salts of the bases, preferably as nitrates 
or sulphates, must be neutral or faintly acid ; excess of 
alkali or acid interferes. Such a solution can be obtained 
by evaporating the filtrate from the guanine in Kossel’s 
method (page 286), and dissolving the residue in nitric 
acid; this is neutralized with sodium carbonate, using 
methyl-orange as indicator. On the addition of excess of 
sodium picrate the adenine is thrown down as a clear yel- 
low flocculent precipitate. If the precipitation is made at 
the boiling temperature, on cooling the adenine salt sepa- 
rates in a crystalline condition and is more easily filtered 
and washed. After standing fifteen minutes the precipitate 
is filtered off by the aid of a suction-pump on a weighed 
filter, washed with cold water, and dried at 100°. As a 
correction for the solubility of the adenine picrate, 2.4 mg. 
per 100 c.c. filtrate can be added to the calculated amount 
of adenine. 
The hypoxanthine picrate is very soluble, and, therefore, 
remains in solution. In this it is estimated according to 
the method described on page 302. 
Adenine, when crystallized from warm or impure solu- 
tions, is obtained either as an amorphous substance, pearly 
plates, or in the form of very small microscopic needles ; 
from dilute cold solutions it separates in long, needle-shaped 
crystals containing three molecules of water. This water 
of crystallization is lost on exposure to the air or on heating 
to 53°, and the crystals become opaque. It is soluble in 
about 1086 parts of water at the ordinary temperature; 
more easily in hot water, from which, on cooling, it recrys- 
tallizes. The aqueous solution possesses a neutral reaction. 
The free base is insoluble in ether, chloroform, and alcohol; 
soluble in glacial acetic acid, and somewhat in hot alcohol. 
It dissolves readily in mineral acids, yielding well-crystal- 
lizable salts. The fixed alkalies dissolve it with ease, but 
on neutralization of the solution it is reprecipitated In 
aqueous ammonium hydrate it is more readily soluble than 
guanine (which is insoluble, Schindler), and more diffi- 
cultly soluble than hypoxanthine—a fact which is made use 288 
BACTERIAL POISONS. 
of to effect a separation from these bases. It is but slightly 
soluble in sodium carbonate. 
Adenine can be heated to 278° without melting; at this 
temperature it becomes slightly yellow, and yields a white 
sublimate. It can be completely volatilized without decom- 
position, by heating on an oil-bath to 220° ; the sublimate 
consists of pure, white, plumose needles of adenine, but at 
250° partial decomposition occurs, and some hydrocyanic 
acid forms. When heated with potassium hydrate to 200° 
on an oil-bath, it yields a considerable quantity of potas- 
sium cyanide. Adenine is quite indifferent to the action of 
acids, alkalies, and even oxidizing agents. Thus, it may be 
boiled for hours with baryta, potash, or hydrochloric acid, 
without suffering decomposition. But when heated with 
dilute hydrochloric acid, or concentrated hydriodic acid, 
in a sealed tube at a temperature exceeding 100°, adenine 
is completely decomposed, with formation of carbonic acid 
and ammonia : 
C5H5N5 + 51I20 + 50 = 5C02 + 5^H3. 
The free base, as well as benzoyl-adenine, is unaffected by 
the weak oxidizing action of potassium permanganate, but 
on stronger oxidation it is wholly destroyed. Bromine water 
produces in aqueous solutions of adenine an oily precipitate, 
which, on contact with potassium hydrate or ammonia, gives 
a beautiful red or violet color. Sodium amalgam and zinc 
chloride appear to have no action; but on boiling with zinc 
and hydrochloric acid it yields a very unstable reduction- 
product, which in the presence of oxygen first assumes a red 
color, and finally throws down a reddish-brown precipitate. 
This brown substance appears to be identical with azulmic 
acid, which has been known for a long time as a product of 
the polymerization of hydrocyanic acid. 
When adenine’is evaporated on the water-bath with dilute 
or fuming nitric acid, it gives a white residue which fails to 
give any coloration with sodium, ammonium, or barium 
hydrate. Similarly, it does not give the so-called Weidel’s 
reaction (murexide test) on evaporation with chlorine water 
and exposure of the residue to an ammoniacal atmosphere. CHEMISTRY OF THE LEUCOMAINES. 
289 
In this respect it resembles hypoxanthine, which, when 
pure, does not answer to either of these tests. Another test 
for adenine, which, however, is given also by hypoxanthine 
but not by guanine and caffeine, is as follows: The sub- 
stance to be tested is digested for half an hour with zinc 
and hydrochloric acid in a test-tube on the water-bath. If 
adenine is present, the solution will assume on standing, 
more rapidly on shaking, a ruby-red coloration, which later 
on turns into a brownish-red. This reaction depends upon 
the formation of a reduction-product, which, owing to its 
unstable nature, is soon oxidized by the oxygen of the 
atmosphere into a brownish, amorphous substance, appa- 
rently identical with azulmic acid. 
On treatment with nitrous acid, it is converted into hypo- 
xanthine according to the equation : 
c6h5n5 + hno2 = c5h4n4o+ n2 + h2o. 
This formation of hypoxanthine from adenine is analogous 
to Strecker’s transformation of guanine into xanthine by 
a similar action of nitrous acid (see Guanine). In both 
oases the change of a highly nitrogenized into a less nitro- 
genized body is accomplished by replacing an Nil group 
by O, or, more exactly, of an NII2 group by OH. In fact, 
the change is identical with that seen in the conversion 
of primary amines into primary alcohols. Thus, 
c2h,nh2 + hno2 = c2h5oh + n2 +h3o. 
Ethylamine. Ethyl Alcohol. 
In the extraction of adenine from the mother-liquors of 
tea-leaves after removal of caffeine, if urea is not added to 
the nitric acid, nearly one-half of the adenine may be con- 
verted into hypoxanthine. By processes of putrefaction 
adenine is converted into hypoxanthine and guanine into 
xanthine (Schindler). The change is, therefore, some- 
what analogous to that produced by nitrous acid. Adenine 
undergoes this decomposition much more rapidly than the 
other xanthine compounds. 
The ease with which adenine and guanine are reduced 
outside of the organism shows that similar changes may take 290 
BACTERIAL POISONS. 
place within the cell-nucleus proper. For we know that 
every cell is endowed with an enormous reducing power, 
and hence it is not difficult to see how the oxygen-free 
adenine can be readily converted into a body or bodies 
which greedily take up oxygen. We must, therefore, look 
upon adenine and guanine not only as the antecedents of 
hypoxanthine and xanthine, but also as intermediate pro- 
ducts which, when they form in the cell, may give rise to 
important chemical processes, especially those of a synthetic 
nature. It is highly probable that the study of the decom- 
position-products of nuclein will explain to us many of the 
metabolic changes in the organism, and throw additional 
light upon the migration of the amido group from the 
proteid molecule to the amido acids and urea derivatives. 
Thus, the formation of xanthine from guanine represents 
the conversion of a guanidine residue into a urea residue. 
A similar change is undoubtedly effected in the transforma- 
tion of adenine into hypoxanthine. 
Adenine unites with bases, acids, and salts. The salts 
of adenine with mineral acids can be recrystallized, thus 
differing from the corresponding salts of guanine and hypo- 
xanthine, which are dissociated by the action of water. 
The solutions of the salts, however, show an acid reaction 
to litmus but not to methyl-orange. 
The hydrochloride, C5H5N5.HC1 + |II20, forms color- 
less, transparent, strongly refracting crystals. One part of 
the anhydrous salt is soluble in 41.9 parts of cold water. 
Microscopically it is distinct from that of hypoxanthine and 
adenine-hypoxan thine. 
The nitrate, CjHgNj.HNOg + l2 II20, crystallizes from 
the aqueous solution in fine, stellate needles. One part of 
the dry salt dissolves in 110.6 parts of water. 
The sulphate, (C5IIvN5)2.H2S04 + 2H20, can be obtained 
from the aqueous solution in two different crystalline forms. 
This may possibly be due to the presence of adenine-hypo- 
xanthine compound (Bkuhns). It is easily soluble in hot 
water, and at the ordinary temperature it is soluble in 153 
parts of water. 
The oxalate, C5H5Na.C2H204 -f H20, is obtained by dis- CHEMISTRY OF THE LEUCOMAINES. 
291 
solving adenine in hot, dilute, aqueous oxalic acid, from 
which solution, on cooling, it separates as a voluminous, 
difficultly soluble precipitate of roundish masses which are 
composed of long, delicate needles. The oxalates of guanine, 
hypoxanthine, and xanthine are more easily soluble than 
that of adenine, and exhibit, moreover, a different appear- 
ance. 
The picrate, C5H5N5.C6H2(N02)30H + H20, is thrown 
down as a bright yellow flocculent precipitate, when aqueous 
solutions of adenine salts are treated with sodium picrate. 
Recrystallized from hot water it forms bright-yellow, very 
voluminous bunches of long fine needles, which, on drying, 
acquire a silky lustre and form a felted mass. It is diffi- 
cultly soluble in cold water (1 : 3500); more readily in hot 
water and in alcohol (96 per cent.); is insoluble in dilute 
acids. The water of crystallization is not lost on exposure 
to air but is driven off at 100° ; the salt then remains un- 
changed even at 220°. A cold concentrated aqueous solu- 
tion of the salt treated with one-tenth its volume of cold con- 
centrated solution of sodium picrate produces a precipitate 
of short fine needles consisting of most of the adenine picrate 
(five-sevenths). The solubility of the picrate can thus be 
reduced to as low as 1 :13750, and on this fact is based the 
quantitative method of Bruhns. The salt can also be 
obtained in its characteristic groups by combining cold 
saturated aqueous adenine solution (1 : 1086) with picric 
acid; with sodium picrate, however, adenine gives no pre- 
cipitate, since the picrate is soluble in an equivalent quan- 
tity of sodium hydrate. Thus is explained Kossel’s 
statement that adenine forms an easily soluble compound 
with picric acid. Heated on a platinum foil it burns slowly 
and leaves considerable carbon residue. The very bright 
yellow color of the salt serves to distinguish it from most 
of the other picrates, especially guanine picrate. 
The platinochloride, (C5H5N5.HCl)2PtCl4, crystallizes 
from dilute aqueous solution in small yellow needles. The 
concentrated aqueous solution of this sail, when boiled for 
some time, decomposes, with the separation of a clear, 292 
BACTERIAL POISON'S. 
yellow powder, which is but slightly soluble in cold water, 
and has the composition C5H5N5.HCl.PtCl4. 
The aurochloride, on evaporation, yields very charac- 
teristic forms. 
The silver salt of adenine, C5H4AgN5, is formed when 
silver nitrate is added in molecular proportion to a boiling 
ammoniacal solution of adenine. An excess of silver 
nitrate produces, in the cold, the compound C5H3Ag2N5 + 
H20, which is converted slowly in the cold, immediately on 
warming, into the other salt, according to the equation : 
2(C5H3Ag2N5 + H20) = 2C5H4AgN6 + AgzO + H20. 
Owing to this instability the two compounds are always 
found together in varying proportion. Both are difficultly 
soluble in water, and ammonia even at the boiling-point. 
The precipitation of adenine by an ammonical silver solu- 
tion is complete, and is therefore available for quantitative 
estimation. 
Adenine silver nitrate, C5H5N5.AgN03, (Ag = 35.4 per 
cent.), corresponds to the similar hypoxanthine and guanine 
salts. It is obtained by dissolving the above silver com- 
pounds in hot nitric acid; and from this solution, on cool- 
ing, it separates in needle-shaped crystals, which are not 
permanent. This lack of stability, as compared with the 
permanent hypoxanthine silver nitrate, was first pointed 
out by Kossel, and was thought to be due to loss of nitric 
acid in washing, and also by heating at 100°. Bruhns, 
however, has shown that the acidity of the wash-water is 
indicated by litmus, but not by methyl-orange, which is 
not colored red by silver nitrate. The reaction is, there- 
fore, due not to free nitric acid, but to silver nitrate. It 
would seem that adenine, as well as hypoxanthine, and pos- 
sibly xanthine, form silver compounds containing one and 
two molecules of silver nitrate; the greater the quantity of 
silver nitrate used the higher is the per cent, of silver, i.e., 
the more of the latter compound is formal. These are very 
unstable, and are decomposed by dilute nitric acid, more so 
by water, into silver nitrate and the compound containing 
one molecule of silver nitrate. We have in this behavior CHEMISTRY OF THE LEUCOMAINES. 
293 
an interesting case of mass-action and chemical equilibrium 
between adenine, silver nitrate, nitric acid and water. 
Ammonium hydrate removes the nitric acid from this as 
easily as from the hypoxanthine compound, and there is 
formed, according to the composition of the original salt, a 
varying mixture of C5H4AgN5 and C5H3Ag2N5 -f- H20. 
The solubility in nitric acid is about the same as that of 
hypoxanthine silver nitrate. 
Adenine silver picrate, C5H4AgN5.C6H2(NO2)30H + 
H20, is obtained as an amorphous voluminous yellow pre- 
cipitate when silver nitrate is added to a cold aqueous solu- 
tion of adenine picrate. If the latter solution is previously 
raised to the boiling-point the precipitate then soon becomes 
crystalline and rapidly subsides. The adenine can thus be 
almost wholly removed from solution. The crystalline 
form loses its water of crystallization at 120°, while the 
amorphous form does not appreciably decrease in weight and 
its composition does not appear to be as constant as that of 
the corresponding hypoxanthine compound. On treatment 
with ammonium hydrate the picric acid is removed, and 
adenine silver, C5H4AgN5, is left, stained yellow by traces 
of picric acid. 
Adenine-mercury picrate, (C5H4N5)2Hg.2C6H2(N02)3OII, 
can be prepared by treating a hot concentrated aqueous 
solution of adenine picrate with an excess of sodium picrate 
and then with mercuric chloride. It forms a yellow gran- 
ular crystalline precipitate (microscopic needles) which rap- 
idly subsides and increases in quantity as the solution cools. 
Its composition apparently varies, containing one to two 
molecules of water, according to the temperature of the 
solution. One molecule is given off at 100°, and the second 
at 105°-120°. The latter preparation, then, on exposure to 
the air, rapidly absorbs one molecule of water. The ob- 
ject of the sodium picrate in the precipitation is to combine 
with the hydrochloric acid, which is set free. The precipi- 
tate produced by mercuric chloride in cold adenine picrate 
solution shows yellow and white granules, and is not homo- 
geneous. Bruhns considers it to be a mixture of the aden- 
ine-mercury picrate and the compound C5H4N5Hg2Cl3; if 294 
BACTERIAL POISONS. 
sodium picrate is added, however, the pure adenine-mercury 
picrate forms, since no hydrochloric acid is set free. 
Adenine-mercuric chloride, C8H4N8HgC], is thrown down 
as a white, finely granular precipitate when a boiling aque- 
ous adenine solution is treated gradually with concentrated 
mercuric chloride solution. It is formed according to the 
following reaction : 
C5H5N5 + HgCl2 = C6H4N6HgCl + HC1. 
That free hydrochloric acid forms can be ascertained by 
methyl orange. Treated with ammonium hydrate the 
chlorine is removed, and there is formed apparently the 
compound C5H4N5HgOII. If dissolved in warm dilute 
hydrochloric acid and allowed to crystallize, the double salt 
C5II5X5.11 Cl. HgC 12 -f- 2II20 separates in long stellate silky 
needles. 
Another mercury compound, C5H4]Sr5Hg2Cl3, is obtained 
when the precipitation takes place in the cold. The 
precipitate is white, flocculent, and anhydrous. In this 
reaction, as above, for each adenine molecule an equivalent 
of hydrochloric acid is set free. This same body is also 
produced when an adenine solution is boiled with a large 
excess of mercuric chloride and as little hydrochloric as 
possible to effect solution. On cooling small stellate needles 
separate out, which do not lose their weight at 110°. It 
can also be obtained by boiling the following compounds 
with water. 
When adenine is boiled with a large excess of mercuric 
chloride and much hydrochloric acid to completely dissolve 
the precipitate that first forms, there is deposited on cooling 
a crystalline product, which is variable in its composition, 
and apparently consists of double salts of adenine and 
mercuric chloride, such as C5H5N5.HC1.5HgCl2 aud C,H5NV 
HC1.6HgCl2. On boiling with water these rapidly de- 
compose, forming the compound C5H4N5.Hg2Cl3. The 
formation of a double salt, C5H5N5.HCl.HgCl2 + 2H20 
is described above. 
Adenine-mercury cyanide, (C5H5N5)2IIg(CN)2, separates CHEMISTRY OF THE • LEUCOMAINES. 
295 
as stellate needles and plates when a mixture of hot solu- 
tions of adenine and mercuric cyanide are allowed to cool. 
An adenine bismuth iodide, C. HrN5.H1.2Bif3 2H20, 
is obtained when an aqueous adenine solution is treated 
with potassium bismuth iodide containing free hydriodic 
acid. The heavy precipitate, which in color resembles 
carbon monoxide haemoglobin, consists of microscopic glit- 
tering red needles. On contact with much water it partly 
decomposes, forming light reddish-yellow amorphous floc- 
cules, which become darkish-brown at 100°. 
Adenine bromide. By treating well-dried adenine with 
excess of dried bromine a dark-red body is obtained which 
appears to contain six atoms of bromine. On mere ex- 
posure to the air, more rapidly on heating at 100°-120°, 
it decomposes, yielding bromine and a brom-adenine, 
C5H4Bi\N5. This compound is white, difficultly soluble in 
cold water (1 : 10,000), more readily in hot water, very 
easily in ammonia. It crystallizes from water or dilute 
ammonia in stellate needles. It is a rather strong base and 
forms well-characterized salts from which it is thrown 
down as a white micro-crystalline precipitate by addition 
of ammonia. It is also formed from adenine-bromide by 
treatment with sodium bisulphite. The picrate resembles 
that of adenine but is more voluminous; silver compounds 
are also formed resembling those of adenine. The silver 
nitrate compound decomposes on boiling with nitric acid 
with separation of silver bromide. It is only difficultly 
attacked by boiling alcoholic potash. 
When adenine is treated with zinc and hydrochloric acid, 
in the cold, it forms a difficultly soluble crystalline double 
salt which has not been obtained in the pure state. This 
double salt is not obtained by direct treatment of adenine 
hydrochloride with zinc chloride. 
One of the hydrogen atoms of adenine is capable of re- 
placement by organic radicals. Thus it forms crystalline 
methyl and ethyl compounds. 
Acetyl-adenine, C5H4N5.CO.CH3, can be obtained by 
heating the anhydrous base with an excess of acetic anhy- 
dride for some time, in an oil-bath, at 130°. It crystallizes 296 
BACTERIAL POISONS. 
in small white scales which dissolve but slightly in cold 
water and in alcohol; more readily in hot water, in dilute 
acids and alkalies. Heated to 260° it becomes yellow but 
does not melt. 
Benzoyl-adenine, C5H4N5.CO.C6H5, is obtained by the 
action of benzoic anhydride, but not of benzoyl chloride, 
on adenine. It crystallizes from water in long, lustrous, 
thin needles which sometimes are grouped in bundles, and 
melt at 234°-235°. It is easily soluble in hot alcohol, from 
which it recrystallizes on cooling; also in dilute acids and 
in ammonia. With ammoniacal silver nitrate it gives a 
precipitate resembling that of adenine, but is more readily 
soluble in ammonia. This compound is quite stable, since 
it decomposes very slowly on boiling with hydrochloric 
acid; on protracted boiling with water it is changed into 
adenine and benzoic acid. 
Benzyl-adenine, CsH4Ns.CH2.CfiH5, was obtained by 
Thoiss by heating well-dried adenine with benzyl chloride 
to boiling (178°) on an oil-bath. The compound forms 
pure white microscopic crystals and melts at 259°. It is 
easily soluble in hot water and in hot alcohol. With acids 
it forms salts from which alkalies throw down the base. 
The hydrochloride forms fine glossy needles which are 
readily soluble in alcohol and in water, but not in ether. 
The sulphate and nitrate possess similar properties. Like 
adenine it yields a silver compound which is insoluble in 
ammonia. On reduction with zinc and hydrochloric acid 
it forms an amorphous red unstable compound. Treated 
with nitrous acid, benzyl-adenine is reduced to benzyl-hypo- 
xanthine, thus showing that the benzyl group replaces a 
hydrogen atom in the group C5H4H4, which Kossel has 
called adenyl (see page 307). 
Benzyl-adenine picrate, C12HnN5.CfiH2(N02)30H, is ob- 
tained as fine felted yellow needles, which are fairly soluble 
in water and in alcohol; insoluble in ether. 
A methyl-adenine was obtained by Thoiss in an impure 
state by heating dried adenine with methyl iodide in a sealed 
tube at 100°. It can be crystallized from absolute alcohol. 
The aqueous solution of the base is precipitated by baryta CHEMISTRY OF THE LEUCOMAINES. 
297 
water; alcoholic zinc chloride also yields a precipitate which 
is soluble in excess of ammonium hydrate. Mercuric nitrate 
also gives a precipitate. Cadmium chloride yields a pre- 
cipitate which dissolves on warming, reappears on cooling, 
and is soluble in ammonia. Basic lead acetate has no effect. 
Nothing definite is known in regard to the physiological 
action of adenine, except that when fed to dogs it appears 
to be eliminated as such, in part at least, by the urine. 
Adenine-Hypoxanthine, 06H5N5 -f- C5H4N40. The 
occurrence of this compound was observed by Kossee, but 
it was isolated and studied for the first time by Bruhns. 
It can be prepared by cooling a hot aqueous solution of 
equal parts of the two bases. At first it is obtained as 
thick, starch-like semi-transparent masses, which later in 
part become white and chalky. By spontaneous evapora- 
tion of its solution in very dilute ammonia it forms pearly 
aggregates of very small radially arranged needles, which 
contain water of crystallization. These effloresce some- 
what and lose the water at 100°. The compound is more 
readily soluble in water than its components, but an exact 
determination of its solubility is impossible, inasmuch as 
the separation from hot solutions is not completed for some 
weeks. Any adenine present can be separated by recrys- 
tallization. It forms a distinct crystalline hydrochloride, 
which should be borne in mind when examining microscopic- 
ally for the two bases; but the combination is loose, since 
addition of gold chloride brings down the characteristic 
gold salt of adenine. Ordinarily it does not form salts 
with sulphuric or nitric acids, but more often is decomposed 
by these, so that the difficultly soluble adenine crystallizes 
out. Once, however, Bruhns obtained a sulphate which 
differed from the pure adenine and hypoxanthine sulphates ; 
thus is perhaps explained the observation of Kossel that 
adenine sulphate forms crystals belonging to two systems. 
The compound can be decomposed into its constituents by 
fractional crystallization of the sulphate or nitrate; but 
better by forming the picrates, which are very unequally 
soluble in water. The existence of this compound undoubt- 298 
BACTERIAL POISONS. 
edly explains many of the mistakes and discrepancies con- 
cerning the properties of hypoxanthine, which it resembles 
more than adenine, and for the same reason, perhaps, 
adenine was so often overlooked. 
Hypoxanthine, C5II4N40, sometimes also known as 
sarcine or sarkine, was discovered by Scherer (1850) in 
splenic pulp and in the muscles of the heart, and was 
named thus because it contains one atom of oxygen less 
than xanthine. It has since been obtained, usually accom- 
panying adenine and guanine, from nearly all of the 
animal tissues and organs rich in nucleated cells, i. e., in 
nuclein. It has been found in blood after death, but not 
in blood when flowing through the bloodvessels. Salomon 
lias recently shown it to be a normal constituent of urine, 
present, however, in an exceedingly minute quantity. In 
the blood and urine of leucocytlnemic patients it occurs in 
increased quantity owing to the abnormally large number 
of nucleated white blood-corpuscles in circulation (see page 
284). Bence Jones observed in the urine of a boy, who 
about three years before showed the symptoms of renal 
colic, a deposit of characteristic whetstone-like crystals, 
resembling uric acid, but differing from the latter by dis- 
solving readily on the application of heat, while from 
hydrochloric acid it crystallized in elongated six-sided 
plates. These crystals he believed to be those of xan- 
thine, but Scherer and others consider them to be hypo- 
xanthine. It is therefore quite possible, though very rare, 
for this base to form a deposit in the urine and to be 
confounded in shape with uric acid. Thudichum has 
obtained it from the urine of persons sick with liver or 
kidney diseases. 
Among other places it has been found in the brain, 
muscle, serum, marrow of bones, kidney, heart, spleen, 
liver, peripheral muscles (sarkine of Strecker) ; in the 
spawn of salmon (Piccard), in the testicles of the bull 
(Salomon), in the nuclein of pus and red corpuscles (Kos- 
Sel), in developing eggs, and in putrefaction of albumin 
(Salomon). It has also been found in the spores of lyco- 299 
CHEMISTRY OF THE LEUCOMAINES. 
podium, and in the pollen of various plants, in seed of black 
pepper, in grass, clover, oats, bran of wheat, larvae of ants ; 
in the juice of potato (Schulze) ; in certain wines 
(Kayser) ; in the aqueous decoction of yeast of beer 
(Schutzenberger) ; and also in the liquid in which yeast 
is grown (Bechamp). Demant has shown it to be rela- 
tively abundant in the muscles of pigeons in a state of in- 
anition, while in muscles of well-fed pigeons it is said to 
be entirely absent. Salomon found hypoxanthine and 
xanthine in the cotyledons of lupine, as well as in the 
sprouts of malt, while Reinke and Rodewald observed 
these two bases together with guanine in JEthalium sep- 
ticum—with adenine, xanthine, and theophylline, it occurs 
in tea-leaves (Kossel). 
Hypoxanthine has not been extracted from the pancreas, 
where it seems to be replaced by guanine, or rather by 
adenine. It seems that hypoxanthine bears a relation to 
adenine similar to that which we see between glycocoll and 
glycocollic acid. 
Hypoxanthine occurs frequently in plants together with 
the other members of this group, namely, adenine, guanine, 
and xanthine. The widely distributed character of these 
bases is due to the presence of a parent substance, viz., 
nuclein, the necessary constituent of all cells capable of 
development, which under the influence of acids, and 
probably likewise of ferments, decomposes into the above- 
mentioned bases. They may, therefore, be considered as 
the first steps in the retrograde metamorphosis of all 
tissues, since they have their origin in nuclein, an impor- 
tant proteid substance. Recent advances in biological 
chemistry have shown that the undeveloped eggs of various 
insects and birds yield much less quantity of xanthine 
bodies (hypoxanthine, xanthine, etc.) on treatment with 
dilute acid than the partially developed eggs (Tichomiroff, 
Kossel). This is dependent upon the remarkable fact 
observed by Kossel that the nuclein of undeveloped 
chicken eggs differs from the nuclein of cell nuclei and 
resembles that obtained from milk. For, while the nuclein 
from the cell nuclei decomposes into adenine, guanine, 300 
BACTERIAL POISONS. 
hypoxanthine, etc., that from undeveloped eggs and from 
milk yields no nitrogenous bases on treatment with acids. 
But as the egg develops, i.e., the nucleated cells increase in 
number, this latter nuclein is gradually converted or gives 
way to the ordinary cell nuclein, and hence it is that the 
chick embryo yields guanine, hypoxanthine, and possibly 
adenine. 
Unquestionably, the presence of hypoxanthine, etc., in 
developing cells is due to the presence of the nuclein mole- 
cule, from which it is readily split olf. In muscle, however, 
hypoxanthine and xanthine appear to exist preformed, and 
bear no relation to nuclein, since they are in the free condi- 
tion, and can be extracted from the tissue by water. For 
an explanation of this peculiar fact, see Adenine, page 284, 
and Guanine, page 308. 
According to the observations of Salomon and Chit- 
tenden, hypoxanthine is formed by the digestion of blood 
fibrin with gastric juice, pancreatic juice, or on heating with 
water or dilute acids. Egg albumin under the same con- 
ditions does not yield any hypoxanthine, except when 
treated with pancreatic juice. These observations require 
repetition, inasmuch as the fibrin used undoubtedly con- 
tained nuclein, which, as we now know, readily decomposes 
under those conditions into its characteristic nitrogenous 
bases. Be that as it may, it appears, however, to be one 
of the products formed by the decomposition and succes- 
sive oxidation of proteid matter previous to the formation 
of uric acid and urea. 
When a mixture of guanine, xanthine, and hypoxanthine 
is allowed to putrefy, the bases decompose and disappear in 
the order named. Hypoxanthine resists bacterial action 
the longest, and this corresponds with its behavior to re- 
agents (Baginsky). Adenine during putrefaction, in the 
absence of air, is converted into hypoxanthine, and guanine 
is correspondingly changed into xanthine (Schindler). 
An imido group is, therefore, replaced by oxygen, and 
probably goes to form urea. This conversion is a very 
important fact, since the process of putrefaction, as Hoppe- 
Seyler has repeatedly pointed out, is analogous to the CHEMISTRY OF THE LEUCOMAINES. 
301 
vital process, and the same chemical change may take place 
in the animal organs. The same change very probably 
takes place in the auto-digestion of yeast. Its formation 
under these conditions can be represented thus : 
C5H5N5 + H2Q = C5H4¥40 + NH3. 
Hypoxanthine can be readily obtained from a number of 
closely related substances. Thus, carnine, by the action of 
oxidizing agents, is converted into hypoxanthine (page 328). 
For this reason Weidel and Schutzenberger regard 
hypoxanthine as derived from carnine, but this view is now 
entirely set aside by our present knowledge of the relation 
of this base to nuclein. 
Again, it can be obtained from adenine (page 289) by the 
action of nitrous acid. The relation that hypoxanthine 
bears to uric acid is shown by the fact that the latter is 
converted by nascent hydrogen first into xanthine, and 
finally into hypoxanthine. 
C5H4N403 + 2H2 = C5H4N40 + 2HzO. 
This transformation of uric acid into hypoxanthine is of 
especial importance, since together with Horbaczewski’s 
synthesis of uric acid, accomplished by acting on urea with 
either glycocoll or trichlorlactamide, it constitutes the last 
step in the complete synthesis of hypoxanthine from its 
elements. 
Hypoxanthine has been hitherto regarded as a step lower 
than guanine in the series of nitrogenous products of 
regressive metamorphosis, and consequently was considered 
as derived from guanine. The investigations of Kossel, 
however, show that it arises not from guanine but from 
adenine. On the other hand, guanine is to be looked upon 
as the source of xanthine. It is probable that in the 
organism it is oxidized as soon as it is set free from the 
nuclein, forming successively xanthine, uric acid, urea, etc., 
and the small quantity present in the urine is all that has 
escaped oxidation. When fed to dogs, it was observed that 
the amount of hypoxanthine present in the urine decreased, 
Uric Acid. 302 
BACTERIAL POISONS. 
and even became less in amount than before the experiment; 
but, on the other hand, the amount of xanthine in the urine 
was found to have been increased above the normal. This 
shows that hypoxanthine in the body is oxidized probably 
first to xanthine, then into uric acid. According to Robert 
hypoxanthine is a true muscle stimulant. 
The fact that hypoxanthine is so widely distributed in 
the organism, and in much larger quantities than was 
formerly supposed, shows that it constitutes, together with 
the closely related bodies creatine, xanthine, guanine, etc., 
the normal antecedents of urea and uric acid. This view is 
furthermore strengthened since hypoxanthine is especially 
abundant in those organs which are most active in pro- 
ducing metabolic changes in the body, viz., the liver and 
spleen. 
It may be prepared from the urine, according to the 
method given under paraxanthine (page 322); or from 
extract of meat, or from glandular organs, such as the liver, 
spleen, etc., by the process on page 285. Nuclein, on de- 
composition with acids, yields about one per cent, of this 
base. It can be determined with adenine indirectly by 
Schindler’s method (page 286); but better still directly 
by Bruhn’s picrate method (see page 286). After the 
adenine has been precipitated by sodium picrate, the deter- 
mination of hypoxanthine in the filtrate is not difficult if 
hydrochloric and other acids, the silver salts of which do 
not quite dissolve in ammonia, are absent. The filtrate 
from the adenine picrate is rendered slightly alkaline with 
ammonia and precipitated with silver nitrate at the boiling- 
point. The slightly yellow-colored precipitate is washed 
with hot water till the wash-water is colorless; then dried 
at 120° for from two to three hours, when it has the com- 
position 2C5H2Ag2N40 + H20. It contains, however, traces 
of picric acid and some adenine silver, and hence the quan- 
tity of hypoxanthine calculated from the weight obtained is 
higher than it really is. Bruiins, as a correction, subtracts 
3.0 mg. from the calculated quantity of hypoxanthine. 
A more convenient method than the preceding is to esti- 
mate hypoxanthine as hypoxanthine silver picrate. The chemistry of the leucomaines. 
303 
filtrate from the adenine picrate (page 287) is raised to the 
boiling-point and silver nitrate solution gradually added. 
The precipitate is washed with cold water till the wash- 
water is colorless, then dried at 100°, when its composition 
is represented by the formula C5H3AgN40.C6H2(NO2)3OH. 
The calculated quantity of hypoxanthine here is likewise 
slightly higher than it should be. Bruhns deducts 1.0 
mg. from the calculated result. 
In the presence of hydrochloric acid, etc., the deter- 
mination of hypoxanthine is somewhat circuitous since the 
precipitated silver chloride must be separated from the 
hypoxanthine compound. The best procedure in this case 
is to saturate the filtrate from adenine picrate with am- 
monia and precipitate it completely with silver nitrate. 
The precipitate is washed with hot water (a thorough wash- 
ing is not necessary), then it is boiled several times with 
nitric acid of 1.1 specific gravity. The acid each time is 
rapidly decanted on to a small filter, and finally the residue 
washed on the filter with 10 c.c. of the hot acid (total 100 
c.c.). To the combined acid filtrate silver nitrate is added, 
and the whole set aside for twenty-four hours. The pre- 
cipitate is dried at 100°. The amount of hypoxanthine 
lost depends upon the quantity of silver chloride present. 
The correction to be added is 3.1 mg. (Bruhns). In 
Neubauer-Kossel’s method the mixed adenine and hypo- 
xanthine silver salts can be decomposed with a little hydro- 
chloric acid and estimated in this way. 
Hypoxanthine is a white, colorless, crystalline powder, 
sometimes in part amorphous ; according to Bruhns, pure 
hypoxanthine does not form floccules and bunches of micro- 
scopic needles, but usually coherent crusts, which consist of 
roundish, sharp-cornered granules; some resemble quadratic 
octahedra. It is soluble in about 300 parts of cold water 
(Strecker). The base separates slowly from aqueous 
solutions, and when pure the solubility, even in the begin- 
ning, is less than 1: 300. At the end of four days Bruhns 
found it to be 1 :1880. It is more easily soluble in boiling 
water (78 parts), and, on cooling, separates in the form of 
white, crystalline floccules, thus differing from xanthine, 304 
BACTERIAL POISONS. 
which is amorphous. The solubility in cold alcohol is very 
slight, about 1 :1000. It dissolves in acids and alkalies 
without decomposition, and from solutions in the latter 
it can be precipitated by passing carbonic acid, or by the 
addition of acetic acid. The aqueous solution possesses a 
neutral reaction. The free base can be heated up to 150° 
without suffering decomposition, but above this temperature 
it sublimes, and partially decomposes, with evolution of 
hydrocyanic acid. When heated with potassium hydrate 
to 200°, it yields ammonia and potassium cyanide. Heated 
with water to 200°, it decomposes into carbonic acid, formic 
acid, and ammonia, and in this respect it agrees with adenine 
(page 288). The properties of Strecker’s sarkine agree 
closely with those of adenine-hypoxanthine ; and, inasmuch 
as the latter has been often described as hypoxanthine, it is 
very desirable that the properties of hypoxanthine be re- 
determined. 
When evaporated with an oxidizing agent, chlorine water 
or nitric acid, the residue is said to give on contact with 
ammonia vapors a rose-red color (Weidel, nmrexide test). 
Kossel, however, has shown that this is due to the presence 
of xanthine, and that pure hypoxanthine does not give either 
the murexide test or the xanthine reaction. According to 
Strecker, concentrated nitric acid converts hypoxanthine 
into a nitro-compound, which in turn, by the action of a 
reducing agent, is changed into xanthine. This statement 
has not been confirmed either by Fischer or by Kossel. 
It does not give a green color with sodium hydrate and 
chloride of lime—distinction from xanthine (page 316). 
With acids it yields crystallizable compounds, and, like 
the amido acids, it forms compounds with bases, and also 
with metallic salts, such as silver nitrate and copper acetate. 
The hydrochloride, C5H4N4O.IICl -f- H20, crystallizes in 
needles, and, like the nitrate and sulphate, it is dissociated 
on contact with water. The crystalline form is character- 
istic and distinct from that of adenine, as well as adenine- 
hypoxanthine. The nitrate forms thick prisms or roundish 
masses, readily soluble in water and ammonia. Platinum CHEMISTRY OF THE LEUCOMAIJSTES. 
305 
chloride forms a yellow, crystalline double salt, having the 
composition C5H4H4O.HCl.PtCl4. 
The picrate forms yellow prisms easily soluble in water, 
which solution is not affected as that of adenine by sodium 
picrate. 
Hypoxanthine silver, C5H2Ag2N4O.H20. All attempts 
to obtain a compound containing but one atom of silver 
in the molecule, corresponding to the adenine compound 
C5H4AgH5, have failed. The above compound was first 
prepared by Strecker, and given the formula C5H4N40. 
Ag20 ; but the former is preferable, since on heating at 120° 
two and a half molecules of water are lost and 
2C5H2Ag2N40 + H20 (Ag = 60.2 per cent.) results. 
At 140°-150° it loses again in weight and becomes gradu- 
ally gray; on exposure to air it absorbs moisture. In this 
form hvpoxanthine can be estimated quantitatively (see 
page 302); the presence of sodium picrate does not interfere, 
but chlorides, etc., do. It is insoluble in hot water. The 
compound, C5H2Ag2N40.3lI20, is obtained in the form of 
microscopic needles, by treating pure hypoxanthine silver 
nitrate with excess of aqueous ammonia. On boiling with 
ammonia-water it is but slightly dissolved, and appears to 
slowly lose a part of its water of crystallization. As a 
result of the decomposition one-half of the hypoxanthine 
passes into solution and can be recovered on boiling with 
addition of silver nitrate in the crystalline form ; or in the 
cold, as the usual amorphous precipitate, C.112 Ag2N40.H20. 
Hypoxanthine silver nitrate, C,II4X4().AgN03, (Ag = 
35.29 per cent.), is the best-known compound ; its formula 
was established by Strecker. It is obtained by dissolving 
the above precipitate, produced by addition of silver 
nitrate to an ammoniacal solution of the base, in hot nitric 
acid, specific gravity 1.1 ; on cooling the hypoxanthine 
silver nitrate crystallizes in the form of tufts of microscopic 
needles or plates. Heated at 100°-120° it remains con- 
stant in weight; the quantity of silver present, when deter- 
mined, is always somewhat higher than the theoretical, 306 
BACTERIAL POISONS. 
especially if an excess of silver nitrate is employed in the 
precipitation. The explanation of this fact is probably 
that given under Adenine, though presence of silver chlo- 
ride may partly be the cause. On treatment with am- 
monia it loses not only nitric acid but also half of the 
hypoxanthine, and C5H2Ag2N40.3H20 forms. The change 
takes place readily even in the cold, and if during the 
digestion an excess of silver nitrate is added, the hypo- 
xanthine set free is converted into this compound, which is 
wholly constant in composition compared with the hypo- 
xanthine silver nitrate. The conversion is quantitative. 
Very dilute hydrochloric acid, as well as hydrogen sulphide, 
removes the silver from this compound. 
Hypoxanthine- si lver picrate, 
C5II3AgN40.C6H2(N02)30H (Ag = 22.88 per cent.), 
is gradually formed by adding silver nitrate to a boiling 
solution of hypoxanthine picrate. The precipitate is granular 
and of a lemon-yellow color, and consists of aggregations of 
line short needles. It is slightly soluble in hot, insoluble in 
cold water. It is, therefore, applicable for a quantitative 
determination of the base. Aqueous ammonia very readily 
and completely removes the picric acid from the compound, 
and the residue is hypoxanthine silver, which is slightly 
colored yellow by a trace of picric acid ; half of the hypo- 
xanthine passes into solution. Nitric acid with difficulty 
converts it into hypoxanthine silver nitrate. 
Hypoxanthine mercuric chloride, CsII3N4OHgCl, is ob- 
tained by adding an equivalent quantity of mercuric chloride 
to a boiling solution of hypoxanthine. The precipitate, 
which increases on cooling, is crystalline. 
A second compound, C5II3N4OIIg2Cl3, is produced by 
adding a strong excess of mercuric chloride, in the cold, to 
an aqueous solution of hypoxanthine. It forms a heavy 
granular micro-crystalline precipitate, which contains some 
water of crystallization. 
By boiling the preceding compound with just sufficient 
hydrochloric acid to effect complete solution, there is formed 
on standing a precipitate of white roundish aggregates of 307 
CHEMISTRY OF THE LEUCOMAIJSTE3. 
leafy or needle-shaped glittering crystals which have the 
composition C5H4N4OHgCl2 + H20. 
The following table of Bruhns illustrates the analogy 
existing between the mercury compounds of adenine and 
hypoxanthine and similar derivatives of ammonium : 
Ammonium. Adenine. Hypoxanthine. 
NH2HgCl 05H4N5HgCI C5H3N40HgCl(-f-H20) 
NH2Hg2Cl3 C6H4N5Hg2Cl3 C5H3N40Hg2Cl3(+H20) 
(NH,)lHg01,{(g^§(20^s(NOi)sOH C5H1N.OHgCl,(+H,0) 
A brom-hypoxanthine compound corresponding to that of 
adenine lias not been obtained. 
Benzyl-1 lypoxanthine, C5H3N4O.CH2.C6H5, was obtained 
by Tiioiss by the action of nitrous acid on benzyl-adenine. 
It forms a white crystalline mass which under the micro- 
scope consists of thin plates. It is easily soluble in hot 
water, dilute alcohol, and in acetic ether; insoluble in ether 
and chloroform. It melts at 280°. It appears, as Kossel 
has pointed out, that adenine and hypoxanthine contain a 
group, C5H4N4, which he named adenyl. The formation of 
the benzyl derivatives of these two bases shows that the 
hydrogen atom which is replaced occurs in the adenyl and 
not in the imido group. According to this view adenine is 
to be considered as adenylimide (C5H4N4.NH) and hypo- 
xanthine as adenyloxide (C5H4N40). 
Phosphomolybdic acid precipitates hypoxanthine from 
acid solution, and in general it gives the ordinary alkaloidal 
reactions. 
It is not precipitated by ammoniacal basic lead acetate. 
Copper acetate does not precipitate it in the cold, but does 
ou boiling. This fact has been made use of in the isolation 
of hypoxanthine. Mercuric chloride, as well as mercuric 
nitrate, produces a flocculent precipitate. 
Altogether, in its behavior to reagents it resembles xan- 
thine to a very considerable degree. The two can be 
separated, however, by the different solubilities of the 
hydrochlorides in water, and more especially of the silver 
salt in nitric acid. 
Physiological Action.—25-100 mg, begin to act on frogs 308 
BACTERIAL POISONS. 
in from six to twenty-four hours, and produce increased re- 
flex excitability and convulsive attacks ; 5-100 mg. is fatal 
(Filehne). When injected subcutaneously into hepato- 
tomized geese or chickens a corresponding increase in uric 
acid secretion is observed (v. Mach). This conversion is 
analogous to that observed by Stadthagen in the case of 
guanine (page 310), and shows that in the xanthine bodies 
we have antecedents of uric acid apart from the synthesis of 
the latter from ammonia in the liver. The process by which 
this change is effected is undoubtedly one of oxidation. 
Guanine, C5H5N60, was discovered, in 1844, by Unger, 
as a constituent of guano, in which it is present in varying 
quantities according to the region from which the guano 
comes. Thus, the Peruvian guano is reported as containing 
the largest proportion of this base, and on that account this 
variety is employed when it is desired to prepare guanine. 
Since its discovery by Unger, it has been met with in a 
very large number of tissues, both animal and vegetable; 
in the liver, pancreas, lungs, retina, in the thymus gland of 
the calf, and in the testicle substance of the bull; in the 
scales of the bleak, and in the swimming-bladder of fish, as 
well as in the excrements of birds, of insects, as the garden 
spider, in which it occurs with a small quantity of uric acid 
(Weinmann), and is to be regarded as a decomposition 
product of proteids formed in the tissues of the spider. It 
is also found in the spawn and testicle of salmon, and 
Schulze and others have shown it to be present in the 
young leaves of the plane-tree, of vine, etc., also in grass, 
clover, oats, as well as in the pollen of various plants. 
Schutzenberger has isolated it, together with hypoxan- 
thine, xanthine, and carnine, from yeast which had been 
allowed to stand in contact with water at near the body- 
temperature. Pathologically, it occurs in the muscles, liga- 
ments, and joints of swine suffering from the disease known 
as guanine-gout. Normally, guanine, like adenine, is 
present in muscle tissue only in traces. It has never been 
found in the urine, though xanthine has been mistaken for 
guanine by some. 309 
CHEMISTRY OF THE LEUCOMAINES. 
As to the origin of this sulstance in the organism very 
little has been known up to within a few years, exetpt so 
far as it has been shown to be, together with other members 
of this group, a transitory procluet in the retrograde meta- 
morphosis of nitrogenous foods and tissues. In the ease of 
the lower animals it is evidently the end-proeluet of all 
change, inasmuch as it is excreted as such. Our knowledge 
as to the immediate origin of this and the other allied bases 
has lately been extended by the brilliant researches of 
Kossel on the decomposition products of nuclein, in which 
he has shown that this essential constituent of all nucleated 
cells, whether animal or vegetable, decomposes under the 
action of water or dilute acids into adenine, guanine, hypo- 
xanthine, and xanthine. We know that the first two bases 
are readily converted by the action of nitrous acid into the 
other two; that is to say, an NH group in these bases is 
replaced by an atom of 0—a change which it is not at all 
unlikely takes place in the tissues, perhaps in every cell 
nucleus. That such a change is quite probable is shown by 
the putrefaction experiments of Schindler, whereby aden- 
ine and guanine were converted respectively into hypoxan- 
thine and xanthine. If this explanation is correct, then 
adenine and guanine are transition-products between the 
complex proteid molecule on the one hand, and hypoxan- 
tliine and xanthine on the other. These two, in turn, form 
the connecting link to the last step in the regressive meta- 
morphosis of the nitrogenous elements of the tissues, viz., 
the formation of uric acid and urea. We can thus trace a 
succession of cycles from the complex nuclein molecule, 
which is apparently indispensable to the functional activity 
of all reproductable cells, to the physiologically waste pro- 
ducts urea and uric acicl. 
Schulze and Bosshaed recently (1886) found in young 
vetch, clover, ergot, etc., a new base, to which they have 
given the name vernine. It has the formula 
and is of especial interest at this point, since on heating 
with hydrochloric acid it apparently yields guanine. We 
have, therefore, at least two well-defined sources of guanine, 
the nucleins and vernine. 310 
BACTERIAL POISONS. 
Neither adenine nor guanine oecur in normal muscle 
further than in mere traces, a fact which can only be 
explained on the ground that the muscle tissue is poor in 
nucleated cells, and hence in nuclein. Just as the muscle 
cell has become morphologically differentiated from the 
typical cell, it may be looked upon also as having under- 
gone a concomitant chemical differentiation, inasmuch as we 
no longer find the phosphoric acid, xanthine, and hypo- 
xanthine in the same chemical combination as they occur in 
the original cell. The phosphoric acid, instead of existing 
as a part of an organic compound, is present in the muscle 
tissue as a salt; similarly the hypoxanthine and xanthine 
occur in the free condition, extractable by water, and no 
longer in combination with other groups of atoms consti- 
tuting a part of a more complex molecule—nuclein. 
Guanine and creatine apparently mutually replace one 
another. Thus, in the muscle, as just stated, guanine occurs 
only in traces, whereas creatine is especially abundant. 
This may find its explanation in the fact that both are sub- 
stituted guanidines. Creatine is regarded by Hoppe- 
Seyler as an intermediate product in the formation of 
urea, and a similar role, it will be remembered, belongs to 
guanine. From Stadt ha gen’s experiments on dogs we 
know that guanine ingested, produces an increase in the 
amount of uric acid and nrea excreted, and the same is 
also true of the nuclein derived from yeast. These results 
have led him to the conclusion that in mammals uric acid 
is a direct, or more or less altered cleavage product of pro- 
teids, notwithstanding the fact that in birds it is the result 
of synthesis in the liver. 
In the decomposition of nuclein-containing substances, 
such as yeast, liver, spleen, etc., by dilute acids, neither 
adenine nor guanine is found alone, but they are always 
accompanied by hypoxanthine, and usually by a very small 
quantity of xanthine. 
Guanine may be readily prepared from Peruvian guano 
by boiling it repeatedly with milk of lime until the liquid 
becomes colorless. The residue, consisting largely of uric 
acid and guanine, is boiled with a solution of sodium car- CHEMISTRY OF THE LEUCOMAINES. 
311 
bonate, filtered, and the filtrate, after the addition of sodium 
acetate, is strongly acidulated with hydrochloric acid. This 
precipitates the guanine, together with some uric acid. The 
precipitate is dissolved in boiling hydrochloric acid, and the 
guanine then thrown out of solution by the addition of am- 
monium hydrate. Guanine is also obtained in the decom- 
position of nuclein with dilute acids, and can, therefore, be 
prepared from such cellular organs as the spleen, pan- 
creas, etc., according to the method given on page 285. It 
should be noted here that in the decomposition of the mixed 
silver compounds with hydrogen sulphide or ammonium 
sulphide (Schindler) the guanine, often only in part, passes 
into solution with adenine and hypoxanthine, and the re- 
mainder is held back in the silver sulphide precipitate. The 
latter should, therefore, be boiled with dilute hydrochloric 
acid, and on saturating the filtrate with ammonia the guan- 
ine after a while separates. That portion of the guanine 
which did pass into solution with the other two bases is 
separated from them by digestion with ammonia on a water- 
bath. The two portions are then combined, transferred to 
a filter, previously dried at 110° and weighed, washed well 
with ammonia, then dried and weighed. 
The free base forms a white, amorphous powder, insol- 
uble in water, alcohol, ether, and ammonium hydrate; 
easily soluble in mineral acids, fixed alkalies, and in excess 
of concentrated ammonium hydrate. It can be heated to 
above 200° without undergoing decomposition. When 
evaporated with strong nitric acid it gives a yellow residue, 
and tin’s on the addition of sodium hydrate assumes a red 
color, which on heating becomes purple, then indigo-blue; 
on cooling it returns to a yellow, passing through purple 
and reddish-yellow shades due, according to V. Brucke, to 
absorption of water. This is the so-called xanthine reac- 
tion, and is supposed to be due to the formation of xanthine 
and a nitro product. It is given best by guanine, then by 
xanthine, and is not given by either hypoxanthine or 
adenine. 
Nitrous acid converts it directly into xanthine, thus : 
C5H5N50 + HN02 = C5H4N402 + n2 + ii2o. 312 
BACTERIAL POISONS. 
This reaction is identical with that of adenine, whereby 
hypoxanthine is formed (see page 289). By putrefaction in 
the absence of air it forms xanthine (Schindler). The 
change can be represented by the equation : 
c5H5N5o + H2o = c5h4n4o2 + nh3. 
On oxidation with potassium permanganate it yields urea, 
oxalic acid, and oxy-guanine. By hydrochloric acid and 
potassium chlorate it is oxidized to carbonic acid, guani- 
dine, and parabanic acid, according to the equation : 
CO-NH. TT 
C5H5N50 + H20 + 30 = | >C0 + >C = NH -f C02. 
CO—NH7 
According to Strecker, a small amount of xanthine is 
formed in this reaction, and it is quite possible that this 
base is also formed on oxidation with nitric acid. 
Guanine combines with acids, bases, and salts. It unites 
with bases to form crystalline compounds ; and with one or 
two equivalents of acid it also yields crystallizable salts. 
Thus, with hydrochloric acid it forms the two salts, 
C5H5N50.(HC1)2 and C5HsN,O.HC1 + H20. Similar com- 
binations can be obtained with nitric acid. The sulphate 
(C5H5N50)2H2S04, crystallizes in long needles, and, like 
the other salts, is decomposable by water. The platino- 
chloride, (C6H,N5O.HCl)2PtCl4 4- 2H20, is readily obtained 
in a crystalline condition. The silver compound is 
soluble in hot nitric acid, and on cooling recrystallizes 
in fine, needle-shaped crystals, having the composition 
C5H5N60.AgNO3. 
The solutions of the hydrochloride are precipitated by 
mercuric chloride and nitrate, potassium chromate, potas- 
sium ferricyanide, and by picric acid. Basic lead acetate 
gives a precipitate only on addition of ammonium hydrate. 
The reaction with picric acid (Capranica) is said to be 
very characteristic, and a means of distinguishing this base 
from xanthine and hypoxanthine. It is best obtained by 
adding a cold, saturated solution of picric acid to the warm 
Parabanic Acid. 
Guanidine. CHEMISTRY OF THE LEUCOMAINES. 
313 
acidulated solution of guanine, when a light, crystalline 
precipitate forms. Under the microscope it appears in 
pencil-shaped, fern-like tufts of fine, orange-yellow needles. 
Physiologically guanine like uric acid is inert (Filehne). 
Xanthine, C5H4N402, is also very widely distributed in 
the organism, and lias been met with in almost all the 
tissues and liquids of the animal economy. Together with 
hypoxanthine, guanine, and possibly adenine, it occurs in 
many plants, among which may be mentioned lupine, 
sethalium, sprouts of malt, tea-leaves (Baginsky), auto- 
digestion of yeast, gourd seeds, soja beans, etc. It was 
first discovered by Marcet (1819) in a urinary calculus, 
and since then has been frequently found as the only or 
chief constituent of many calculi. Unger and Phipson 
have extracted it from guano, while Salomon has shown 
it to be one of the products formed in the pancreatic diges- 
tion of fibrin. Schutzenberger found it together with 
carnine and hypoxanthine in the liquors from yeast. It is 
a normal constituent of the urine, but is present only in 
extremely minute quantities. During the use of sulphur- 
baths, or after the thorough application of sulphur salves, 
the quantity of xanthine in the urine is considerably in- 
creased. It is likewise more abundant in the urine of leuco- 
cythsemic patients, for the reasons already given on page 
283. Baginski holds that the amount of xanthine nor- 
mally present in the urine may be increased tenfold in the 
case of acute nephritis. Bence Jones observed in the 
urine of a child sick with renal colic, a deposit of crystals 
which he considered to be xanthine, but other observers 
are inclined to regard the crystals as those of hypo- 
xanthine. Vaughan has reported the presence of xan- 
thine in deposits from the urine of patients with enlarged 
spleen. 
Xanthine may be prepared synthetically in several ways. 
Thus, it may be obtained by the reduction of uric acid by 
means of sodium amalgam, according to the equation : 
c,h4n4o3 + H2 = c,h4n4o2 + H20. 
Uric Acid. Xanthine, 314 
BACTERIAL POISONS. 
Now that uric acid has been prepared synthetically, this 
forms the final step in the complete synthesis of xanthine. 
By further action of nascent hydrogen the xanthine in turn 
is converted into hypoxanthine. The reverse operation, 
the conversion of hypoxanthine into xanthine, though re- 
ported by Strecker has not been confirmed by Fischer 
or by Kossee. It is, therefore, evident that these bodies 
form a continuous oxidation series with uric acid as the 
final product. Although this change is unquestionably the 
one which goes on in the animal economy, yet all attempts 
to reproduce it in the laboratory by oxidation with potas- 
sium permanganate or nitric acid have apparently yielded 
only negative results. Again, xanthine may be prepared 
from guanine by putrefaction of the latter, or by oxidation 
with nitrous acid. The change may be represented by this 
equation : 
c5h5n5o + hno2 = c5h4n4o2 + n2 + h2o. 
Guanine. 
Xanthine. 
This reaction, first described by Strecker (1858), corre- 
sponds exactly to the one by which Kossel has transformed 
adenine into hypoxanthine (see page 289). 
Gautier, starting out on the hypothesis that xanthine 
is a polymerization-product of hydrocyanic acid, has en- 
deavored to prepare it directly from this compound. In- 
deed, he claims to have succeeded in effecting the synthesis 
of not only xanthine, but also its homologue, by simply 
heating hydrocyanic acid in a sealed tube with water and a 
little acetic acid, the latter being added to neutralize any 
ammonia that might form. He expresses the reaction as 
follows : 
11HCN + 4H20 = C5H4N402 + C6II6N402 + 3NH3. 
Nearly all of the methods that have been employed for 
the preparation of xanthine are based upon its precipitation 
as the insoluble silver compound. From the urine it can 
be isolated according to the method given under paraxan- 
tliine, on page 322. It may also be obtained from the 
Xanthine. 
Methyl-xanthine. CHEMISTRY OF THE LEUCOMAINES. 
315 
urine by Hofmeister’s method. The urine, acidulated 
with hydrochloric acid, is precipitated with phosphotungstic 
acid; the precipitate is decomposed by warming with 
baryta, filtered, and the filtrate is freed from barium by the 
cautious addition of sulphuric acid. The solution is then 
made alkaline with ammonium hydrate, any traces of phos- 
phates that appear are filtered oifi, and finally it is precipi- 
tated by addition of ammoniaeal silver nitrate. The pre- 
cipitate which forms consists of the silver compounds of the 
xanthine bodies, and is purified by dissolving in hot nitric 
acid, as given on page 285. Xanthine has been shown to 
be formed at the same time with guanine, adenine, and 
hvpoxanthine, in the decomposition of nuclein by means of 
dilute acids. It may, therefore, be prepared from cellular 
organs according to the method given under Adenine. The 
method of its preparation from tea-leaves is also given 
elsewhere. 
Xanthine is a white, granular, amorphous body, and is 
deposited from hot aqueous solution on cooling in colorless 
floecules, or as a fine powder, which, under the microscope, 
is seen to consist of rounded granules. When occurring in 
calculi, it forms compact, moderately hard, yellow or brown 
fragments, which, on being rubbed with the finger-uail, 
assume a wax-like appearance. It is difficultly soluble in 
cold water (about 14,000 parts), alcohol, and ether; some- 
what more soluble in boiling water (about 1200 parts). It 
is soluble in alkalies and alkali carbonates, not bicarbon- 
ate, and from these solutions it is precipitated on neutral- 
ization with acids, or by passing carbonic acid. In warm 
ammonia it dissolves more readily than does uric acid or 
guanine, and on cooling the ammonium compound recrys- 
tallizes. It acts as a weak base, and as a weak acid ; with 
salts of the heavy metals it forms difficultly soluble or 
insoluble compounds. Its basic properties, however, are 
weaker than those of hypoxanthine or guanine. 
When xanthine is evaporated with nitric acid it leaves a 
lemon-yellow residue (hence its name), which is not changed 
by ammonium hydrate—distinction from uric acid—but 
with potassium hydrate becomes yellowish-red, on heating 
purple-red. When added to a mixture of bleaching powder 316 
BACTERIAL POISON'S. 
and sodium liydrate in a Avatch-glass the solution becomes 
covered by a dark-green scum, which changes to a brown, 
and soon disappears—distinction from hypoxanthine. 
By means of a very interesting synthetic reaction, xan- 
thine may be converted into theobromine, the active con- 
stituent of Theobroma cacao. Tims, the xanthine is dissolved 
in a sufficient quantity of sodium hydrate, necessary to form 
the neutral compound CsH2Na2N402, and this product, 
when treated with boiling acetate of lead, yields a white 
precipitate of lead xanthine, C5H2PbN402. This is dried 
at 130°, then heated for twelve hours at 100° with methyl 
iodide, when the dimethyl derivative, C6H2(CH3)2N402, is 
formed. This compound is identical with the natural theo- 
bromine, and by a similar treatment is converted into tri- 
methyl-xanthine or caffeine. The relation of xanthine to 
theine (caffeine) is shown in the fact that it exists together 
with hypoxanthine, adenine, and possibly guanine, iu fresh 
tea-leaves. It is, therefore, clear, that by starting from 
guanine of guano we can produce successively xanthine, 
dimethyl xanthine, and trimethyl xanthine, the last two 
compounds being identical with the alkaloids of theobroma 
and of coffee. 
Nascent hydrogen converts this base into hypoxanthine, 
but the reverse operation, the oxidation of hypoxanthine 
into xanthine, has been questioned of late by Kossel and 
others. On heating, a small portion volatilizes; the 
greater part decomposes into ammonium carbonate, cyan- 
ogen, and hydrocyanic acid. Heated to 200° with hydro- 
chloric acid, it decomposes with the formation of ammonia, 
carbonic acid, formic acid, and glycoeoll (E. Schmidt). 
When bromine is allowcel to act on xanthine, there is 
formed a substitution compound, having the formula 
C5H3BrN402. With potassium chlorate and hydrochloric 
acid it yields alloxan and urea. 
Xanthine is a weak base, which dissolves in acids with 
the formation of salts. 
The hydrochloride, CSH4N402.HC1, is difficultly soluble 
in water, more so than the corresponding salt of hypoxan- 
thine, from which it is deposited in glistening six-sided CHEMISTRY OF THE LEUCOMAINES. 
317 
plates, often forming aggregations. Its solution does not 
precipitate platinum chloride. The nitrate forms fine yellow 
crystals. 
The sulphate, C5H4H402.H2S04 + H20, crystallizes in 
microscopic glistening rhombic plates, decomposable by 
water. 
With baryta water xanthine forms the difficultly soluble 
compound C5H4H402.Ba(OH)2, which corresponds to the 
hypoxanthine salt C5H4lSr4O.Ba(OH)2, and to that of 
guanine. 
From ammoniacal solution, silver nitrate precipitates the 
compound C5H4N402.Ag20, which is insoluble in ammonia, 
but soluble in hot nitric acid. From the nitric acid solu- 
tion, on long standing, there separates the compound 
C8H4N402.AgN03, which, on contact with water, decom- 
poses, giving off nitric acid. The ammoniacal solution is 
also precipitated by lead acetate—separation from hypo- 
xanthine—also by calcium and zinc chlorides. Cupric 
acetate gives a precipitate only on boiling. The aqueous 
solution is not precipitated by lead acetate, but is by phos- 
phomolybdic acid, phosphotungstic acid, by mercurous and 
mercuric salts. Picric acid gives an easily soluble com- 
pound, which resembles that of hypoxanthine, but differs 
from that of guanine. 
As to the physiological relation of xanthine very little 
need be said. It bears the same relation to guanine that 
hypoxanthine does to adenine, and, like the latter, is to be 
looked upon as an intermediate compound, a step lower 
than guanine, and nearer the limit of oxidation—uric acid. 
It is quite probable that in the body it is oxidized about 
as rapidly as it is formed. Like hypoxanthine, it is to 
be regarded as a true muscle stimulant, especially of the 
heart. (Baginski). According to Filehne it produces 
in frogs a decided muscular rigor and paralysis of the spinal 
cord. The heart muscle is also affected, which is not the 
case with caffeine or theobromine. The fatal dose is less 
than one-half pro mille. In its action it is stronger than 
theobromine, while caffeine is weaker than either of the 
two. Pasohkis and Pal hold that the reverse is true. 318 
BACTERIAL POISONS. 
In closing the description of the preceding bodies it may 
be well to present briefly our present knowledge as to their 
constitution. Gautier, starting out with the idea that 
they are polymerization-products of hydrocyanic acid, has 
deduced theoretically cyclic formulae, recalling the hexagon 
of the benzole derivatives. These formulae, though formid- 
able in appearance, are a complete failure so far as they are 
expressions of chemical reactions. Tims, the formula of 
guanine: 
N= CH 
H —CO —N\g_c)c = NH 
% 
HN NH 
fails to show either a urea or a guanidine residue, and yet 
it is a well-known fact that guanine on oxidation yields 
parabanic acid and guanidine (page 312). In a similar 
manner, his xanthine formula fails to show up the urea 
residues which we know to be present. 
Horbaczewski’s synthesis of uric acid has thrown con- 
siderable light upon the constitution of these bases. As a 
consequence of his method of synthesis uric acid was shown 
to possess the structural formula given below. E. Fischer 
has found, as a result of experimental work, the constitu- 
tion of xanthine to be expressed by the subjoined formula. 
We know that uric acid on treatment with nascent hydro- 
gen is converted into xanthine, then into hypoxanthine. 
It follows, therefore, that a relation exists between hypo- 
xanthine and xanthine similar to that between xanthine 
and uric acid. The formula of hypoxanthine, as deduced 
from this relation, and given below, probably represents 
its constitution quite closely. It is possible, however, that 
the CH and CO groups will be found to occupy the 
reverse position which they are given in this formula, in 
which case corresponding changes must be made in the 
formulae of guanine and adenine. The latter two are based 
upon the relation which these bodies bear to xanthine and CHEMISTRY OF THE LEUCOMAINES. 
319 
hypoxanthine, and cannot be said to be the result of direct 
experimental evidence. 
NH—C —NH N = C —NH N=C — NH 
I I I 
CO . CO CH 
I I 'll 
NII—C CO NII—C CO N—C CO 
II II I II I 
CO—NH CII—NH CH—NH 
c5h4n403 c5ii4n4o2 c5h4n4o 
N = C —NH N=C — NH 
I I 
CO CH 
I II 
NH—C C=NH N—C C=NII 
II I II I 
CH—NH . CH—NH 
Uric Acid. 
Xanthine. 
Hypoxanthine. 
Guanine. 
c5h5n50 
Adenine. 
C5H5N5- 
Heteroxanthine, C6H6N402, is a new base which was 
isolated from the urine in 1884 by Salomon. In its 
composition it is methyl-xanthine, and is intermediate 
between xanthine and paraxanthine or dimethyl-xanthine. 
It occurs in the urine of man and of the dog in about the 
same amount as paraxanthine, and the method for its isola- 
tion will be found under the description of that base. It 
is a remarkable fact that tin’s base occurs in dog’s urine 
unaccompanied by paraxanthine, and the same seems to 
hold true for the urine of leucocythaemic persons. Salomon 
examined the liver and muscles of a dog, but was unable 
to obtain any heteroxanthine or paraxanthine, and the total 
amount of xanthine bodies present was about normal. 
Hence, he is inclined to think that these two bases may 
possibly have their origin in the kidney. Unlike the other 
xanthine bodies, heteroxanthine has not as yet been isolated 320 
BACTERIAL POISONS. 
from plants, meat extract, or guano. The amount of 
xanthine bodies present in the urine is unaffected by phos- 
phorus poisoning. Neither this base nor paraxanthine lias 
been found in bull’s testicles; xanthine is also absent, and 
only hypoxanthine and guanine were found to be present. 
Heteroxanthine forms a white amorphous powder, which 
sometimes on prolonged contact with water forms micro- 
scopic crystalline tufts. It is very difficultly soluble in 
cold water; much more easily in hot water, and the solu- 
tion thus obtained is neutral in reaction. It is easily soluble 
in ammonium hydrate, but is insoluble in alcohol and ether. 
When heated it volatilizes without melting and at the same 
time gives off a small quantity of hydrocyanic acid. On 
evaporation with nitric acid on the water-bath (xanthine 
reaction) it remains as a pure white residue, which on con- 
tact with sodium hydrate develops only a trace of reddish 
coloration or none at all. Weidel’s test (page 328) pro- 
duces a splendid red color, which becomes blue on the ad- 
dition of sodium hydrate. Simple evaporation with chlo- 
rine water gives a similar though not so strong a color 
reaction. 
Silver nitrate produces in ammoniacal, as well as in 
nitric acid solutions, a precipitate which readily dissolves 
on warming in even very dilute nitric acid ; from this 
solution, if not too concentrated, the heteroxanthine silver 
nitrate compound crystallizes in well-formed plate-like 
prismatic crystals. Copper acetate produces in the cold, in 
solutions of heteroxanthine, a clear-green precipitate. It 
is also precipitated by phosphotungstic acid, and by ammo- 
niacal basic lead acetate. Picric acid does not give a yellow- 
colored precipitate in solutions of the hydrochloride. Mer- 
curic chloride readily precipitates heteroxanthine in the 
form of a grayish-yellow compound, which on standing 
twelve to twenty-four hours becomes converted into pure 
white crystalline aggregations. This mercuric compound 
can be converted directly into the corresponding silver 
compound by the addition of silver nitrate and ammonia, 
as described under paraxanthine. 
The hydrochloride is characterized by its rather difficult 321 
CHEMISTRY OF THE LEUCOMAINES. 
solubility and ready crystallization (a distinction from the 
paraxanthine salt). The salt forms large colorless tufts 
of crystals, which on contact with water soon lose their 
transparency and become opaque ; gradually their crystal- 
line form disappears, till finally they completely decom- 
pose with the formation of heteroxanthine. This decom- 
position is hastened by warming, either with or without 
addition of ammonia. Platinum chloride produces in the 
hydrochloric acid solution a precipitate of crystalline double 
salt. 
This base resembles paraxanthine in its property of 
yielding a difficultly soluble precipitate with the fixed 
alkali. This reaction is best brought about by dissolving 
the heteroxanthine hydrochloride in warm dilute sodium 
hydrate, when, on cooling, the corresponding sodium salt 
will crystallize out in oblique-angled plates. These crystals 
dissolve easily in water, and on neutralization of the 
solution with an acid a dense pulverulent precipitate of 
heteroxanthine forms. It can thus be distinguished from 
paraxanthine, the sodium compound of which, on similar 
treatment, yields the characteristic crystalline form of the 
free base. This sodium reaction, therefore, distinguishes it 
at once from xanthine, hypoxanthine, guanine, and para- 
xanthine. It differs from the latter, as has already been 
indicated, in the solubility and amorphous character of the 
free base; in the behavior of the hydrochloride and the 
sodium compound, and in the not giving a precipitate with 
picric acid, nor the characteristic odor given by paraxan- 
thine on heating. 
In its composition, heteroxanthine is, as has already 
been stated, methyl-xanthine and probably is related to if 
not identical with an isomeric body obtained synthetically 
by Gautier (see page 314). The fact nevertheless re- 
mains, that in the urine we have normally a homologous 
series of xanthine bodies, namely, xanthine, heteroxanthine, 
and paraxanthine. 
l\vraxanth ine, C7II8N402, was isolated in 1883 by 
Salomon, who lias since shown it to be a constituent of 322 
BACTERIAL POISONS. 
normal urine, although present in exceedingly minute 
quantity. Thus from 1200 litres of urine, only 1.2 grammes 
(0.0001 per cent.) of tins substance were obtained. It has 
not been found in the urine of dogs or in that of leuco- 
cythsemic patients. Thudichum was the first to isolate 
paraxanthine from the urine, and he named it urotheo- 
bromine (1879). 
The method employed for the isolation of this base is, 
with a slight modification, that of E. Salkowski, as 
originally proposed for the preparation of xanthine bases 
from urine. The urine in portions of 25 to 50 litres is 
made alkaline with ammonium hydrate and allowed to 
stand twenty-four hours. The clear supernatant fluid is 
decanted from the precipitate of phosphates and treated 
with silver nitrate (0.5 to 0.6 gramme per litre). The gray- 
ish precipitate of xanthine compounds which forms is trans- 
ferred to a filter and washed with water till free from 
chloride; it is then suspended in water and decomposed 
with a current of hydrogen sulphide. The liquid is filtered 
by decantation aud the filtrate is evaporated to dryness; 
the residue is extracted with 3 per cent, sulphuric acid to 
remove uric acid; the solution thus obtained, after it lias 
been rendered alkaline with ammonia, is precipitated by 
silver nitrate. 
A better procedure is to concentrate the filtrate directly 
over the flame or on the water-bath, till the uric acid begins 
to crystallize out. It is then filtered, and the filtrate, after 
diluting somewhat with water, is rendered alkaline with 
ammonium hydrate in order to precipitate any remaining 
uric acid and phosphates. The whole is allowed to stand 
one or two days, then filtered, and the filtrate again pre- 
cipitated with silver nitrate. The thoroughly washed pre- 
cipitate of the xanthine compounds, now free from uric 
acid, is dissolved in as little as possible of hot nitric acid 
of specific gravity 1.1, to which a little urea lias been 
added, and the clear solution is set aside for twenty-four 
hours. The silver salt of hypoxanthine crystallizes from 
the solution and is filtered oft*. It can be purified by re- 
peated recrystallization from hot nitric acid, containing a CHEMISTRY OF THE LECCOMAINES. 
323 
little urea, then decomposed with hydrogen sulphide, and 
the filtrate, rendered alkaline with ammonium hydrate, is 
concentrated to a small volume. On standing, pure hypo- 
xanthine crystallizes out. The filtrate from the silver salt 
of hypoxanthine on being rendered alkaline with ammonium 
hydrate gives a precipitate which formerly was regarded as 
consisting entirely of the xanthine silver compound, but 
which from the investigations of Salomon, has been shown 
to be a mixture of the salts of xanthine, paraxanthine, 
and heteroxanthine. 
The separation of these bases is effected by the solubility 
of the free bases in ammonium hydrate. For this purpose 
the precipitate of the mixed silver salts is decomposed with 
hydrogen sulphide, and the filtrate, rendered ammoniacal to 
remove traces of phosphates and oxalates, is moderately 
concentrated. After standing twenty-four hours, heteroxan- 
thine crystallizes out, partly in finely formed sheaves and 
tufts of needles, partly in radially striated masses. The 
fluid is decanted from the crust of heteroxanthine which 
forms in the bottom of the beaker, and after being concen- 
trated somewhat is again allowed to stand. In this way a 
second crop is obtained, and this is repeated till finally the 
separated masses scarcely give a precipitate with sodium 
hydrate. All the heteroxanthine is now united and dis- 
solved in a little hot water by the aid of sodium hydrate. 
After twenty-four hours the greater part of the heteroxan- 
thine crystallizes out in bunches of crystals of sodium 
heteroxanthine, while a small part together with any traces 
of xanthine remains in solution. The crystalline mass is 
dried by pressure, dissolved in a little water, and the solu- 
tion neutralized by addition of hydrochloric acid, when the 
heteroxanthine separates as a pulverulent precipitate. To 
remove any traces of paraxanthine, dissolve in hydrochloric 
acid; on standing forty-eight hours the heteroxanthine salt 
separates, while the easily soluble salt of paraxanthine 
remains in solution. To obtain the pure free heteroxan- 
thine, the hydrochloric salt is evaporated with ammonium 
hydrate; the well-washed residue of heteroxanthine is then 
dissolved in dilute ammonia, the solution filtered, evapor- 324 
BACTERIAL POISONS. 
ated slowly, and the precipitate which forms is finally washed 
with alcohol and ether. 
The original ammoniacal mother-liquors of heteroxan- 
thine yield on further concentration amorphous floccules of 
xanthine, which are removed by filtration ; from the filtrate, 
when concentrated still more, paraxanthine crystallizes out. 
Paraxanthine is obtained in colorless, glassy, generally 
six-sided plates, which are arranged in tufts or rosettes. 
From very concentrated aqueous solutions it crystallizes in 
long, colorless, interwoven needles, which on drying exhibit 
the silky lustre of tyrosin. The crystals belong to the 
monoclinic system, and may crystallize with as well as 
without water. If water is present on careful heating (110°) 
the crystals lose their brilliancy and become whitish and 
opaque, and at 120°-130° the water is completely driven 
off. The conditions under which crystals containing water 
are formed are not known; probably by slow crystalliza- 
tion, whereas rapid crystallization from hot concentrated 
solution yields the anhydrous needles. At about 170°—180° 
sublimation takes place. The melting-point is at about 
284° (Kossel). It can be heated to 250° without melting 
or suffering any decomposition, but when heated more 
strongly it gives off white vapors which possess a distinct 
iso-nitril odor, at the same time it carbonizes and takes fire. 
When evaporated with concentrated nitric acid, as in the 
ordinary xanthine test, it gives only a slight yellow residue. 
On the other hand, Weidel’s test, evaporation with chlo- 
rine water containing a trace of nitric acid, and then placing 
the dry residue into an ammoniacal atmosphere under a 
bell-jar, gives a beautiful rose-red color. 
It is difficultly soluble in cold water (though more easily 
than xanthine); somewhat more readily soluble in hot 
water, and insoluble in ether and alcohol. It is soluble in 
ammonium hydrate, hydrochloric acid, and nitric acid. Its 
solutions are neutral in reaction. 
Silver nitrate produces in nitric acid, as well as in ammo- 
niacal solutions, a flocculent or gelatinous precipitate, which 
in concentrated solutions forms an almost perfect jelly-like 
mass. This silver precipitate is soluble in warm nitric CHEMISTRY OF THE LEUCOMAINES. 
325 
acid, from which on cooling it separates in white crystalline 
tnfts possessing a silky lustre. On decomposition with 
hydrogen sulphide the silver salt yields pure paraxanthine. 
Picric acid produces in the hydrochloric acid solution a 
precipitate consisting of densely felted yellow crystalline 
spangles. 
It is also precipitated by phosphotungstic acid and copper 
acetate; mercuric chloride when added in exeess gives a 
precipitate composed of a mass of colorless prisms, which 
are rather difficultly soluble in cold water ; easily in hot 
water. The crystals of paraxanthine mercuric chloride 
when moderately heated become opaque from loss of water 
of crystallization; at a higher temperature they melt, under- 
going at the same time partial decomposition, and on strong 
heating they evolve disagreeable nauseating vapors. The 
aqueous solution of this mercuric double salt gives with 
silver nitrate an abundant precipitate of silver chloride, 
which disappears on the addition of ammonium hydrate 
and is replaced by the flocculent gelatinous precipitate of 
silver paraxanthine. The hydrochloric acid solution of 
paraxanthine crystallizes with difficulty even when strongly 
concentrated, and on the addition of platinum chloride it 
yields a well-crystallizable orange-colored paraxanthine 
platinochloride. It is not precipitated by basic lead acetate 
nor by mercuric nitrate. 
In its behavior to the xanthine test this base resembles 
hypoxanthine, whereas in giving Weidel’s reaction it 
approaches xanthine. Finally, it coincides with guanine 
by yielding a precipitate with picric acid. Although it 
thus agrees in some of its reactions with all three of these 
xanthine bodies, it can, however, be easily distinguished 
from them by its behavior with the fixed alkalies. Sodium 
or potassium hydrate dissolves these bases and holds them 
in solution, but when added to concentrated paraxanthine 
solution the alkali produces a precipitate of long, glittering, 
crystalline spangles, which under the microscope are seen to 
consist of delicate rectangular, often longitudinally striated, 
plates which are either isolated or united in tufts. Besides 
these crystals there are also present hexagonal plates resem- 326 
BACTERIAL POISONS. 
bling cystin. The crystals are soluble in a little water, or 
on warming, but precipitate again on cooling. Paraxan- 
thine, however, shares with heteroxanthine the property of 
forming a difficultly soluble compound with the fixed alka- 
lies, but can be distinguished from the latter by neutralizing 
with an acid the solution of the sodium or potassium com- 
pound, when, in the case of paraxanthine, there will be 
obtained a precipitate of the characteristic crystals of that 
base ; whereas heteroxanthine is obtained on similar treat- 
ment as a dense pulverulent precipitate. This reaction is 
not given by theophylline. 
It is interesting to observe that paraxanthine is isomeric 
with theobromiue, theophylline, and also with a body re- 
cently described by Fischer as dioxy-dimethyl-purpurine. 
In its composition it is, therefore, a dimethyl-xanthine. 
The physiological action of paraxanthine has been studied 
by Salomon. Injections into the muscles of 1-2 mg. pro- 
duced almost at once a rigor-mortis-like condition of the 
muscles affected, with diminished reflex excitability without 
previous increase; 6-8 mg. introduced into the lymph sac 
brings on a gradual loss of voluntary motion as well as of 
reflex excitability; the rigor is more marked in.the anterior 
extremities, which have a wooden or waxy consistency. 
Dyspnoea is likewise an early symptom, but as soon as rigor 
sets in the respirations drop far below the normal, and may 
even be absent for several minutes. At times the lungs are 
enormously dilated, same as in theobromine. The heart’s 
action is intact till the very last. In mice the reflexes are 
increased almost to a tetanus. The lethal dose for frogs, 
subcutaneously, was found to be 0.15-0.2 per cent, of the 
body-weight—somewhat lower than that of theobromine and 
xanthine. The action of these three bases is very similar. 
They produce in common the slow creeping movements, 
followed by cessation of spontaneous muscle action, com- 
plete loss of reflex excitability without a previous rise, 
and the heart’s action is not affected till in the latest stages. 
Carnine, C7H8N403, was isolated in 1871 from Amer- 
ican meat-extract by Weidel, but has not been obtained CHEMISTRY OF THE LEUCOMAINES. 
327 
from muscle-tissue itself. It lias also been obtained from 
yeast liquors by Sen utze nberger, and from urine by 
Pouchet. It can be separated from the meat-extract, of 
which it forms about one per cent., by the following method 
originally employed by Weidel. The extract is dissolved 
in six or seven parts of warm water, then concentrated 
baryta water is added, avoiding, however, an excess. The 
filtrate is precipitated by basic lead acetate. The precipitate 
is collected, thoroughly washed and pressed, and finally it 
is repeatedly extracted with a large quantity of boiling 
water. The carnine lead salt is thus dissolved out; the 
filtrate, after removal of the lead by hydrogen sulphide, is 
evaporated to a small volume. The concentrated solution 
thus obtained is treated with silver nitrate, which gives a 
precipitate of silver chloride and of the silver salt of car- 
nine. By treatment with ammonium hydrate the silver 
chloride can be completely removed from the precipitate, 
whereas the silver compound of carnine is insoluble in that 
reagent. To obtain pure carnine the silver salt is decom- 
posed with hydrogen sulphide, and the filtrate, after purifi- 
cation by bone-black, is evaporated to crystallization. 
Carnine forms white crystalline masses, which on drying 
become loose and chalk-like. It is very difficultly soluble 
in cold water, easily and completely in boiling water, and 
recrystallizes on cooling. It is insoluble in alcohol and 
ether. The taste is decidedly bitter, and the reaction is 
neutral. The base is not precipitated by neutral lead 
acetate, but is precipitated by the basic salt as a flocculent 
white precipitate, soluble in boiling water. On heating, 
carnine decomposes and takes fire, and at the same time 
gives off a peculiar odor. It crystallizes with one molecule 
of water, which it loses at 100°-110°. 
The hydrochloride, C7H8N403.HC1, is crystalline, and 
decomposes on heating with concentrated hydrochloric acid. 
The platinochloride, C7H8N403.HCl.PtCl4, forms a fine, 
sandy, golden-yellow powder. 
With silver nitrate, carnine unites to form a white floccu- 
lent precipitate, insoluble in nitric acid or in ammonium hy- 
drate. Its formula corresponds to 2(C7II7AgN403)+AgN(33. 328 
BACTERIAL POISONS. 
Carnine is not affected by prolonged boiling with concen- 
trated barium hydrate. Bromine water decomposes it with 
the evolution of gas and the formation of hypoxanthine. 
This change takes place according to the following equa- 
tion : 
C7H8N403 + 2Br = C5H4N4O.HBr + CH3Br + C02. 
A similar decomposition into hypoxanthine is brought about 
by the action of nitric acid, though in this case oxalic acid 
and a yellow body are formed. When carnine is evaporated 
with chlorine water containing a little nitric acid, the resi- 
due, on contact with ammonia, gives a rose-red color 
(murexide test). This is due, according to Weidel, to 
the formation of hypoxanthine, but it has since been shown 
that the latter base does not give this reaction, and hence it 
is due to the production of xanthine, or some similar body. 
The physiological action of carnine has been examined 
somewhat by Brucke, and according to him it is not very 
poisonous. The only effect observed, when taken inter- 
nally, was a fluctuation in the rate of the heart beat, though 
even this was by no means definite in its nature. 
A Base, C4H5N50, was obtained by Gautier from 
fresh muscle tissue of beef, according to the method given 
on page 334, and on account of a resemblance in some of 
its properties with xanthine, he named it pseudoxanthine. 
This name is very inappropriate, not only because it differs 
so much in its empirical formula from that of xanthine, 
C5H4N402, but also because the term pseudoxanthine has 
already been applied by Schultzen and Filehne to a 
body isomeric with xanthine, which was obtained by the 
action of sulphuric acid on uric acid. 
The free base forms a light-yellow powder, slightly 
soluble in cold water, soluble in weak alkali and in hydro- 
chloric acid. The hydrochloride is very soluble, and it 
forms stellate prisms with curved faces, which resemble 
the corresponding salt of hypoxanthine, and to some extent, 
also, the whetstone-shaped crystals of uric acid. 
Like xanthine, its aqueous solution is precipitated in the CHEMISTRY OF THE LEUCOMAINES. 
329 
cold by mercuric chloride, silver nitrate, and by ammo- 
niaeal lead acetate, but not by normal lead acetate. On 
evaporation with nitric acid, the residue gives, on contact 
with potassium hydrate, as in the case of xanthine, a beau- 
tiful orange-red coloration (xanthine reaction). It differs 
from xanthine, not only in its empirical composition, but 
also in its greater solubility, and in its crystalline form. 
It is possible that this base, on account of its great resem- 
blance to xanthine, may have been mistaken, at different 
times, for that compound. 
Gerontine, CsH14N2, is a new base which was isolated 
by Grandis in 1890. It has been repeatedly observed in 
the form of peculiar crystals found in the cell nuclei in the 
liver, particularly of old dogs. The free base is an isomer 
of cadaverine, etc., and resembles it somewhat. It crystal- 
lizes in needles which are readily soluble in water and alco- 
hol ; possesses a strongly alkaline reaction, and yields the 
ordinary alkaloidal reactions. 
The hydrochloride forms prismatic crystals, which are 
deliquescent and easily soluble in alcohol. 
The platinochloride, C5H]4N2.2HCl.PtCl4, is soluble in 
water and crystallizes in spindle-shaped needles, arranged 
in rosettes. It decomposes at 115°. 
The gold salt forms small needles, and is easily soluble 
in water and alcohol. 
It combines with one molecule of mercuric chloride to 
form deliquescent cubes or rectangular prisms containing 
two molecules of water of crystallization. It decomposes 
above 100°. This distinguishes it from cadaverine, which 
combines with three to four molecules of mercuric chloride. 
The crystals observed in the liver are probably the phos- 
phate. 
The new base also yields a benzoyl compound which 
melts at 175°-176°. 
Physiological Action.—It seems to exert a paralyzing 
action upon the nerve centres, and leaves the nerves and 
muscles unaffected. 330 
bacterial poisons. 
Spermine, C2H5N, or CinH2fiN4 (?), is the basic substance 
obtained by Schreiner (1878) from semen, calf’s heart, 
calf’s liver, bull’s testicles, from the organs of leucocythae- 
mics, and also from the surface of anatomical specimens 
kept under alcohol. In 1888 Kunz reported the presence 
of a non-poisonous base, C2H5N, spermine or ethyleneimide 
in cholera cultures. In this case it occurs, then, as a pto- 
maine. A confirmation of the identity of the two bases is 
necessary. Previous to this, however, it had been known 
for a long time under the name of “Charcot-Neumann 
or Leyden crystals,” which are the phosphate of spermine. 
These peculiarly shaped crystals have been found in the 
sputa of a case of emphysema with catarrh, in the bronchial 
discharges in acute bronchitis, as well as in sputa of chronic 
bronchitis, in the blood, spleen, etc., of leucocythsemies and 
ansemics, and in the normal marrow of human bones, as 
well as in human semen. Altogether it seems to have a 
very wide distribution, especially in certain diseases, as in 
leucocythsemia. 
It can be prepared from fresh human semen in the fol- 
lowing manner: The semen is washed out of linen by a 
little warm water, evaporated to dryness, boiled with alco- 
hol, and the insoluble portion is allowed to subside by 
standing some hours. The precipitate is filtered off, washed, 
and dried at 100°. This residue, containing the spermine 
phosphate, is triturated, and then extracted with warm 
ammoniacal water. From this solution, on slow evapora- 
tion, the phosphate crystallizes in its peculiar-shaped 
crystals. 
The free base is obtained, on decomposing the phosphate 
with baryta and evaporating the filtrate, as a colorless 
liquid, which, on cooling, crystallizes. From alcohol it 
crystallizes in wavellite-shaped crystals, which readily 
absorb water and carbonic acid from the atmosphere. 
They are readily soluble in water and in absolute alcohol, 
almost insoluble in ether, and possess a strongly alkaline 
reaction. When heated with platinum it gives off thick, 
white fumes, and a weak ammoniacal odor. With potas- 
sium bismuth iodide it yields orange-colored crystalline CHEMISTRY OF THE LEUCOMAINES. 
331 
floccules, which, under the microscope, appear as long, 
sharp, plumose needles—distinction from diethylenediamine. 
The aqueous solution of the base is precipitated by phos- 
phomolybdic and phosphotungstic acids, tannic acid, gold 
and platinum chlorides. It cannot be volatilized from 
alkaline solution by steam without undergoing decomposi- 
tion (Majert and Schmidt). It is not poisonous. 
The hydrochloride, C2H5N.HC1 (?), crystallizes in six- 
sided prisms, united in tufts, and is extremely soluble in 
water, almost insoluble in absolute alcohol and ether. 
The aurochloride, C2H5N.HC1.AuC13 (?), forms shining, 
golden-yellow, irregular plates, and when freshly precipi- 
tated it is easily soluble in water, alcohol, and ether, but 
the dried salt is incompletely soluble in water. The aque- 
ous solution, treated with magnesium, gives off a sperm- 
like odor. The platinochloride crystallizes in prisms. 
The phosphate, (C2H5N)2.H3P04-|-3II20(?), forms prisms 
and slender double pyramids arranged in rosettes. It is 
difficultly soluble in hot water, insoluble in alcohol, easily 
soluble in dilute acids, alkalies, and alkali carbonates. It 
melts with decomposition at about 170°. It is probable 
that the above formula does not represent the salt as found, 
and from theoretical considerations Ladenburg was in- 
clined to think that Schreiner’s phosphate had the com- 
position (C2H5NH)4Ca(P04)2. 
Ladenbtjrg and Abel prepared in 1888 a compound, 
ethyleneimine, which was first supposed to be isomeric with 
spermine. The reaction whereby it is prepared is similar 
to the one by which Ladenburg effected the synthesis of 
piperidine. Ethylenediamine hydrochloride is subjected to 
dry distillation, when it decomposes into ammonium 
chloride and the hydrochloride of the new base. The re- 
action was supposed to be represented by the equation : 
ch2nh2.hci ch2V 
I = I >NH.HC1+NH4C1. 
ch2nh2.hci ch2 
Since then Ladenburg has shown that the boiling-point 
of this compound did not agree with what it should be 332 
bacterial poisons. 
theoretically, if represented by the above formula. A de- 
termination of the vapor density showed that the molecular 
weight was twice that corresponding to the formula given, 
and hence was C4H]0N2. Majert and Schmidt assuming 
spermine to be ethyleneimine, as was apparently shown by 
Ladenburg and Abel’s investigation, attempted to pre- 
pare the latter on a manufacturing scale with the expecta- 
tion that it might be used as a substitute for Brown- 
Sequard’s testicular fluid. They were soon able to show, 
however, that ethyleneimine did not possess the composition 
assigned to it, but that it was identical with Hofmann’s 
diethylenediamine (piperazine), 
2. CH2\ 
\CH2.OH,/ 
This was soon confirmed by Hofmann and by Ladenburg. 
Spermine was then assumed to be identical with piperazine, 
but recently (1891) Majert and Schmidt compared some 
spermine from Schreiner with their own piperazine and 
found the two bases to be distinct, especially with reference 
to the phosphates and the potassium bismuth iodide pre- 
cipitates. 
About the same time (1891) Poehl announced that the 
composition of spermine was more complex than what it 
had been hitherto supposed to be. He ascribed to it the 
formula C10I I26X4. The formula of the platinum salt cor- 
responded to C10H26N,4HCl.2PtCl4; and that of the gold 
salt was represented by C10H26N4.4HC1.4AuCl3. 
From this it would appear that spermine is essentially 
distinct from piperazine. The composition and structure 
of this interesting base must therefore be considered as not 
settled. 
The nuclein of the spawn of salmon has been found 
by Miescher to exist in a salt-like combination with a 
basic substance, to which he applied the name protamine. 
Picard has found it in the same source, together with 
hypoxanthine and guanine, but no xanthine. The formula 
assigned to this base is quite complex, and cannot be con- 
sidered as definitely settled. Analysis of the platino- CHEMISTRY OF THE LEUCOMAINES. 
333 
chloride gave : Pt=24.64, Cl=26.45, N=15.03, 0=22.80, 
H=4.15, 0=6.93. The hydrochloride forms an amor- 
phous, hygroscopic, sticky mass. 
Leucomaines of the Creatinine Group. 
The knowledge of the formation of basic substances 
(ptomaines) during the putrefaction of nitrogenous organic 
matter, led to a series of investigations having for their 
object the isolation of alkaloidal bodies, if such existed, 
from the normal living tissues of the organism. A number 
of compounds possessing alkaloidal properties, such as the 
xanthine derivatives, already described, had been known 
for a long time, although their physiological relation to 
the animal economy was little, if at all, understood. 
Guaresciii and Mosso, in the course of their researches on 
ptomaines, were among the first to direct their attention to 
the possible presence of ptomaine-like bodies in fresh tissues. 
They obtained in those cases where the extraction was 
carried on without the use of acids, only very minute traces 
of an alkaloidal body (possibly choline), and an inert sub- 
stance, methyl-hydantoin, which, although it can scarcely 
be classed as a basic compound, is closely related to creatine, 
and for this reason will be described at the end of this sec- 
tion. Other Italian chemists, as Paterno and Spica and 
Marino-Zuco, had also shown that the normal fluids and 
tissues of the body were capable of yielding substances 
alkaloidal in nature, and these were regarded by them as 
identical with, or similar to, the ptomaines of Selmi. 
Arginine, C6H14N402, is a base obtained by Schulze 
from the conglutin of lupine sprouts, and according to him 
it is related to creatinine and possibly to the leucomaines of 
Gautier. Lysatine, C6H13N302, and lysatinine, C6HnN30, 
are analogous bases, obtained by Drechsel from casein 
(page 242). These three bases can properly be looked upon 
as important sources of the nitrogenous bases found in 
animals and plants. 
Liebreich, in 1869, discovered in normal urine an 
oxidation-product of choline, probably identical with 334 
BACTERIAL POISONS. 
betaine (pp. 249 and 343), and Pouchet, in 1880, announced 
the presence in the same secretion of allantoin, carnine (page 
344), and an alkaloidal base, which, however, was not 
obtained at that time in sufficient quantity to permit a 
determination of its character. Subsequently he succeeded 
in isolating this base as well as another closely related 
body, both of which will be described in their proper 
place. Gautier has been engaged for a number of years 
in the study of the leucomaines occurring in fresh muscle 
tissue, and he has succeeded in isolating several new 
compounds. 
A number of these substances are credited with possess- 
ing an intensely poisonous action, and if such is the ease 
it is very evident that any undue accumulation of such 
bases in the system, resulting from an interference in the 
elimination, may give rise to serious disturbances. The 
amount of these substances present in the daily yield of 
the urine is very small—so small, indeed, that we must 
rather look upon this small quantity as having escaped 
oxidation in the body. It is well known that the living 
tissues possess an enormous oxidizing and reducing power, 
and, according to Gautier, there is constantly going on 
in the normal tissues of the body a cycle of changes—the 
formation of leucomaines and their subsequent destruction 
by oxidation, before they have accumulated in sufficient 
quantity to produce poisonous effects. 
The following method was employed by Gautier in 
his study of the leucomaines of muscle tissue: The 
finely divided fresh beef-meat or the Liebig’s meat extract 
is treated with twice its weight of water, containing 0.25 
gramme of oxalic acid, and one to two c.c. of commercial 
peroxide of hydrogen per litre. The purpose of these 
precautions is to prevent fermentation. At the end of 
twenty-four hours the liquid is raised to the boiling-point, 
then filtered through linen, and the residue is thoroughly 
squeezed. The filtrate is again raised to the boiling-point 
in order to coagulate any remaining albumin, and finally 
filtered through paper. The clear liquid thus obtained is 
evaporated in a vacuum at a temperature not exceeding CHEMISTRY OF THE LEUCOMAINES. 
335 
50°, and the acid syrupy residue is extracted with 99 
per cent, alcohol; the alcoholic extract is in turn 
evaporated in a vacuum, and the residue taken up with 
warm alcohol of the same strength. The filtered alcoholic 
solution is set aside for twenty-four hours, and any deposit 
which forms is removed by filtration; ether (65°) is then 
added as long as a precipitate continues to form, and the 
whole is again allowed to stand for twenty-four hours. 
The ether-alcoholic filtrate from this precipitate is evapo- 
rated first on the water bath, and finally in a vacuum ; 
the slight residue obtained contains a small quantity of 
basic substances possessing an odor of hawthorn. 
The syrupy precipitate produced by the ether partially 
crystallizes on standing; a little absolute ether is then 
added, and after standing several days more the liquid is 
separated by means of an aspirator from the deposit of 
crystals (A). These are first washed with 99 per cent, 
alcohol, and then extracted with boiling 95 per cent, 
alcohol. The alcoholic solution, concentrated by evapora- 
tion, gives, on cooling, a deposit of lemon-yellow-colored 
crystals of xantho-creatinine (B), from the mother-liquor 
of which there separates a crop of new crystals (C). The 
residue of the crystals (A) left after treatment with the 
boiling 95 per cent, alcohol is extracted with boiling water, 
which afterward gives a slight deposit of yellowish-white 
crystals of amphi-creatine (D). The aqueous mother-liquors 
on concentration yield another deposit of orange-colored 
crystals of cruso-creatinine (E). Gautier has, further- 
more, separated three other bases from the mother-liquors 
of the crystals obtained as above. Thus, a base which he 
named pseudoxanthine is stated to have been obtained by 
evaporating the alcoholic mother-liquors of B, D, E (?) in 
a vacuum, taking up the residue with water, and precipi- 
tating the hot solution with copper acetate. The precipitate 
is decomposed with hydrogen sulphide, and the aqueous 
solution, filtered while boiling-hot, yields a deposit of a 
sulphur-yellow poAvder of pseudoxanthine. Thus, by the 
use of alcohol, ether, and water, Gautier, according to his 
statement, has succeeded in obtaining a sharp separation 336 
BACTERIAL POISONS. 
between these bases. The importance of the subject is such 
as to require not only confirmation of the results arrived at 
by Gautier, but also a more detailed and exact study of 
the chemical and physiological behavior of these bodies. 
To the physiological chemist these substances are of 
especial interest because of the possible relation which they 
bear to the formation of creatine and creatinine in the 
muscle. It will be seen that in the leucomaines of this 
group, as well as in those of the uric acid group, hydro- 
cyanic acid plays a very important part in the molecular 
structure of these bases. Just what the function of this 
cyanogen group is, so far as the vital activity of the tissues 
is concerned, we know very little, though recent investiga- 
tions seem to show that the seat of the cyanogen formation 
lies within the nucleated cell, and is intimately connected 
with the functions of the nuclein molecule. 
Cruso-creatinine, C5H8N40, forms orange-yellow crys- 
tals which are slightly alkaline in reaction, and possess a 
somewhat bitter taste. It yields a soluble, non-deliquescent 
hydrochloride crystallizing in bundles of needles; also a 
soluble platinochloride which forms tufts of beautiful, 
slender prisms. The aurochloride is obtained as slightly 
soluble, crystalline grains, and, like the platinum double 
salt, is partially decomposed on heating. It is not precipi- 
tated by acetate of zinc or by mercuric nitrate, but is pre- 
cipitated in the cold by solutions of alum. Zinc chloride 
produces in somewhat concentrated solutions a pulverulent 
precipitate which dissolves on heating, and recrystallizes 
again when it cools. Like xantho-creatinine, it is not thrown 
out of solution by oxalic or nitric acid, and is thus distin- 
guished from urea and guanidine; nor is it precipitated by 
acetate of copper—a distinction from xanthine derivatives. 
Mercuric chloride produces an abundant flocculent precipi- 
tate which on heating partially dissolves, decomposing at 
the same time. Sodium phosphomolybdate gives a heavy 
yellow precipitate, whereas potassium mercuro-chloride and 
iodine in potassium iodide have no effect. Potassium ferri- 
cyanide is not reduced. This base differs in its composition CHEMISTRY OF THE EEUCOHAINES. 
337 
from creatinine by HCN, the elements of hydrocyanic acid, 
but in its crystalline form and alkaline reaction, and some 
other properties, it would seem to be closely related to this 
latter substance. Because of this apparent relationship and 
its golden-yellow color, Gautier named it cruso-creatinine. 
Xantho-creatinine, C5H10X4O, is the most abundant 
of muscle leucomaines. It crystallizes in sulphur-yellow, 
thin spangles, consisting of nearly rectangular plates which 
resemble somewhat those of cholesterin. It is soft and 
talc-like to the touch ; possesses a slightly bitter taste, and 
when dissolved in boiling alcohol it gives off the odor of 
acetamide, though ordinarily in the cold it has a slight 
cadaveric odor. When heated, the substance evolves an 
odor of roast meat, carbonizes in part, and yields ammonia 
and methylamine. The crystals are amphoteric in reaction, 
are soluble in cold water, and can be recrystallized from 
boiling 99 per cent, alcohol. 
It forms a hydrochloride crystallizing in plumose needles, 
and a very soluble platinochloride; the aurochloride crys- 
tallizes with difficulty. Like creatinine, it is precipitated 
by zinc chloride; the yellowish-white precipitate dissolves 
with partial dissociation on warming, and on cooling sepa- 
rates as isolated or stellate groups of fine needles which 
possess the composition (C5H10X4O)2ZnCl2. Silver nitrate 
throws down, in the cold, a flocculent precipitate which 
likewise dissolves on heating, and recrystallizes in needles. 
Mercuric chloride produces a yellowish-white precipitate. 
It is not precipitated by oxalic or nitric acid, nor by potas- 
tassio-mercuric chloride, or iodine in potassium iodide. 
Tannin produces in time a slight turbidity, while sodium 
phosphomolybdate gives a heavy yellowish precipitate. 
This base is distinguished from the members of the uric 
acid group by not giving a precipitate with copper acetate, 
not even on heating. 
On gentle oxidation with potassium permanganate it is 
converted into a black substance insoluble in acids and 
alkalies, and resembling azulmic acid. By treatment with 
recently precipitated mercuric oxide, it yields a substance 338 
BACTERIAL POISON’S. 
which can be recrystallized from boiling 93 per cent, 
alcohol in needles which possess a slight alkaline reaction, 
and forms a slightly soluble, crystalline platinochloride. 
This new substance is precipitated from alcoholic solution 
by the addition of ether, as a mass of beautiful, white, silky 
needles resembling caffeine. These crystals melt at 174° ; 
caffeine melts at 178°. 
Xantho-creatine, given in fairly large doses, is poison- 
ous, producing in animals depression, somnolence, and 
extreme fatigue, accompanied by frequent defecation and 
vomiting. In its general properties this base resembles 
creatinine very much, and it was on account of this resem- 
blance and its yellow color that it was named xantho-crea- 
tinine. This relation becomes especially evident since this 
base appears in the physiologically active muscle at the 
same time with creatinine, constituting sometimes one-tenth 
of the creatinine present. Monari has found this base in 
the aqueous extract of the muscles of an exhausted dog, 
and also in the urine of soldiers tired by several hours’ 
march. He also demonstrated its presence in the urine of a 
dog after previous injection of creatinine. 
Amphi-creatine, C9H19N704, is slightly soluble and 
crystallizes from boiling water in yellowish-white oblique 
prisms, which possess, if any, a slightly bitter taste. 
When heated to 100° it decrepitates somewhat, and at 
110° it becomes opaque white. Potassium hydrate docs 
not decompose it in the cold. Although a weak base, it 
combines to form salts just as the preceding members of 
this group. The hydrochloride is crystalline, and is not 
deliquescent; the platinochloride forms rhombic plates, 
which are soluble in water, but are insoluble in absolute 
alcohol; the aurochloride crystallizes in easily soluble, very 
small, microscopic crystals, which are tetrahedral to hexa- 
hedral in their habit. It is not precipitated by copper 
acetate or by mercuric chloride; nor does it give the 
murexide test, or the xanthine reaction. Sodium phospho- 
molybdate produces a yellow, pulverulent precipitate. In 
its properties it resembles creatine, and indeed Gautier CHEMISTRY OF THE LEUCOMAINES. 
339 
thinks it may be possibly a combination of creatine, 
C4H9N302, and a base C5H]0N4O2, which, it will be seen, 
differs from the former only by a HCN group. This 
second compound, if it really exists, has an analogy in 
cruso-ereatinine, the relation of which to creatinine may be 
expressed by the equation : 
c5h8n4o = c4h7n3o+hcn. 
Crtjso-oreatinine. Creatinine. 
In a similar manner, amphi-creatine may be regarded as 
C9H19N704 = 2C4H9N302+HCN. 
Amphi-creatine. Creatine. 
A Base, C11H24N10O5, was isolated by Gautier from 
the mother-liquors of xantho-creatinine. It crystallizes in 
colorless or yellowish, thin, apparently rectangular plates, 
which are tasteless, and possess an amphoteric reaction. 
The hydrochloride forms bundles of fine needles; the sul- 
phate yields a confused mass of needles; the platinochlo- 
ride is soluble, non-deliquescent, and crystalline. When 
heated with water in a sealed tube at 180°-200°, it gives 
off ammonia and carbonic acid, and is converted into a 
new base, which, however, has not been studied. This 
reaction may be expressed by the equation : 
CuH24N10O6 = 2C5H10N4O2+CO(NH2)2. 
L BE A. 
The urea which at first forms, is, in turn, decomposed, 
thus : 
co(NH2)2+H2o = co2+2NH3. 
It is to be observed that this base differs in composition 
from the following one by IICN, the hydrocyanic acid 
molecule. 
A Base, CJ2H25Nu05, was obtained from the mother- 
liquors of cruso-creatinine, and forms rectangular silky 
plates, resembling those of the preceding base and of 
xantho-creatinine. It forms crystallizable salts. 
These complex bases will require further study in order 340 
BACTERIAL POISONS. 
to elucidate their physiology, and the possible connection 
which they may have with the formation of urea, and of 
the creatinine derivatives already described. 
Methyl-hydantoin, C4H6N202, = CO yjg CCT 
—This substance was obtained by Guareschi and Mosso 
(1883), by extracting fresh meat with 1-1.5 volumes of 
water (without addition of acid), for two hours at 50°-60°. 
The aqueous extract was evaporated on the water-bath and 
the residue was extracted with 95 per cent, alcohol. This 
alcoholic solution, after the alcohol was driven off, was 
taken up in water, filtered, and the aqueous solution was 
first extracted with ether, then rendered alkaline with 
ammonia, and again extracted with ether. The alkaline 
ether extract gave on evaporation a white crystalline residue 
of methyl-hydanto'in. The amount of this substance 
present in flesh appears to be quite variable, since, at times, 
none whatever can be extracted. Albertoni has isolated 
it from dog’s flesh. Previous to its discovery in flesh by 
Guareschi and Mosso, it was known for a long time as a 
decomposition-product of various nitrogenous bases of the 
body. Thus, Neubauer prepared it by heating creatin- 
ine with barium hydrate, while Huppert obtained it by 
fusing together sarcosine with urea. As it occurs in muscle 
it is probably derived from the creatine, though under 
what conditions this splitting up takes place is not definitely 
known. Acetic and lactic acids are incapable of effecting 
this change. At all events, it belongs to the ureides, and 
is intermediate between creatinine, sarcosine, and urea 
Compare the above formula with that of creatinine, p. 226. 
Methyl-hydanto'in forms prisms which are easily soluble 
in water and alcohol, and but slightly soluble in cold ether. 
It melts at 156° (Salkowski) ; at 159°-160° (Guareschi 
and Mosso). Its aqueous solution is slightly acid in reac- 
tion. On strong heating it volatilizes. When fused with 
potassium hydrate it gives off ammonia; it reduces mercuric 
nitrate in the cold. Treated with mercuric oxide it assumes 
an alkaline reaction, and the filtrate on heating yields CHEMISTRY OF THE LEUCOMAINES. 
341 
metallic mercury. With silver oxide it forms pearly lanceo- 
late plates having the composition C4H5N202. Ag. It does 
not give any alkaloidal reactions. 
Undetermined Leucomaines. 
It was shown at qu5te an early period that exhalations 
from animals contain, besides an increased amount of car- 
bonic acid, some organic matter, the nature of which, on 
account of the exceedingly minute quantity in which it 
occurs, has never been satisfactorily determined. Never- 
theless, various observers did not hesitate to ascribe to it 
the ill effects consequent upon breathing impure air, while 
at the same time the carbonic acid formed during respira- 
tion was considered as either entirely inert or as insignifi- 
cant in its action. Thus, respired air from which moisture 
and carbonic acid have been removed, but which still contains 
the organic vapors, has been found to be highly poisonous. 
On the other hand, if the respired air is drawn through 
a red-hot tube to destroy the organic matter, the air thus 
purified is capable of sustaining life even in presence of a 
large percentage of carbonic acid. While it cannot be, 
therefore, doubted that the organic matter of expired air 
plays a most important part in producing the well-known 
noxious effects resulting from breathing confined and vitiated 
air, nevertheless it would seem from experiments made by 
Angus Smith that the increase of even such small quanti- 
ties of carbonic acid in the air, as from 0.04, the normal 
amount present, to 0.1 per cent., is capable of producing 
systemic disturbances characterized by a decrease in the 
pulse-rate and an increase in the rate of respiration. 
Smith is consequently of the opinion that the constant 
lowering of the pulse in impure air occasioned by the pres- 
ence of carbonic acid must have a depressing effect on 
the vitality. Whatever ill effects the carbonic acid may 
produce of itself, it remains certain that this gas is not the 
most potent and most injurious constituent of respired air; 
Leucomaines of Expired Air. 342 
bacterial poisons. 
and the investigations of Hammond, Nowak, Seegen, 
and others, point conclusively to the organic matter as the 
direct and immediate agent which produces those symp- 
toms of sickness and nausea experienced in badly ventilated 
closed rooms. 
Of special importance to the sanitarian and physician is 
the work on the nature and action of the poisonous principle 
of expired air which has been done by Brown-Sequard, 
d’Arsonval, and 14. Wurtz. The first two observers 
found that the vapors exhaled by dogs, when condensed, 
and the aqueous liquid (20-44 c. c.) thus obtained was in- 
jected into other animals, death was produced, generally 
within twenty-four hours. The symptoms observed were 
dilatation of the pupil, increase of heart-beat to 240-280 
per minute, which may last for several days or even weeks, 
while the temperature remains normal; the respiratory 
movements are generally slowed, and usually there is ob- 
served paralysis of the posterior members. Choleraic diar- 
rhoea is invariably present. As a rule, it appears that 
larger doses cause labored respiration, violent retching, and 
contraction of the pupil. A rapid lowering of temperature, 
0.5° to 5°, is sometimes observed. These same symptoms, 
apparently in aggravated form, were obtained when the 
liquid had been previously boiled for the purpose of de- 
stroying any germs that might be present. The appearances 
presented on post-mortem were much like those observable 
in cardiac sytieope. 
The above work has been confirmed, in part, by 14. 
Wurtz, who, by passing expired air through a solution of 
oxalic acid, has obtained besides ammonia a volatile organic 
base which is precipitated by Bouchardat’s reagent and 
by potassio-mercuric iodide. It is said to form a platinum 
double salt crystallizing in short needles, and a soluble 
gold salt. When heated to 100° it gives off a peculiar 
odor. This basic substance may properly be looked upon 
as a leucoma'ine. 
Dastre and Loye and Lehmann and Jessen have 
repeated the above experiments with wholly negative re- 
sults. It is possible that the most highly poisonous sub- CHEMISTRY OF THE LEUCOMAINES. 
343 
stances formed in the body when there is an insufficient 
air-supply are not eliminated in the exhaled air. 
Sewer-air, according to observations made by Odling, 
contains a basic substance which is probably in composition 
a compound ammonia. It contains, however, more carbon 
than methylamine and less than ethylamine. 
It should be remarked that Jackson has (Dec. 1887) 
announced the presence in expired air of quantities of car- 
bon monoxide gas sufficient to produce the ill effects ordi- 
narily attributed to the organic matter. The presence of 
this poisonous gas must first be fully demonstrated before 
it can be taken into account in the consideration of the 
toxicity of air ; certainly, even if present, it cannot explain 
the results obtained by the French investigators as stated 
above. 
According to Ilosva, expired air contains nitrous acid. 
This may possibly be derived from that which is constantly 
being formed in the mouth, probably by the reduction of 
nitrates (Miller). 
Leucomaines of the TJrine. 
A number of basic substances have been isolated at 
different times from the urine, and on that account they 
may be properly classed as leucoma'ines. Thus, Liebreich 
(1869) found in the urine a base which apparently was an 
oxidation-product of choline, and which lias since been 
regarded as identical with betaine. In 1866 Dupre and 
Bence Jones found, among other things in the urine, an 
alkaloidal body which in sulphuric acid solution possessed 
a blue fluorescence (see p. 347). Most of the members of 
the uric acid group of leucoma'ines have been detected in 
the urine and on account of their well-defined nature they 
are described by themselves. In the urine and feces of 
cystinuria Udranszky and Baumann discovered the well- 
known ptomaines, cadaverine and putrescine. For isola- 
tion, see pp. 207 and 208. 
In 1879, Thudichum announced the presence in the 
urine of four new alkaloids, one of which, urotheobromine, 344 
BACTERIAL poisons. 
was subsequently rediscovered by Salomon and named 
paraxanthine (page 321). Another base which was ob- 
tained, namely, reducine, yielded a barium salt which readily 
reduced the salts of silver and mercury. Its formula prob- 
ably corresponds to C12H24N609 or C6HuN304. A third 
alkaloid, parareducine, formed a zinc compound having the 
composition C6H9N3O.ZnO. A fourth base is said to give 
a compound with platinum chloride and to contain an aro- 
matic nucleus (aromine). Besides these four bases Thudi- 
chum describes two other substances which he considers 
basic. These are urochrome, the normal pigment of the 
urine, and creatinine. 
In 1880, Pouchet announced the presence of carnine, 
C7H8N403, and of another base which he subsequently ana- 
lyzed and found to have either the composition C7H12N402 
or C7H14N402. This substance formed deliquescent fusi- 
form crystals, sometimes crystallized in bundles or irregular 
spheres, which possessed a slightly alkaline reaction and 
combined with acids to form crystallizable salts. It was 
soluble in dilute alcohol, almost insoluble in strong alcohol, 
insoluble in ether. The hydrochloride yielded double salts 
with gold chloride, platinum chloride, and mercuric chlo- 
ride. The platinochloride formed deliquescent golden- 
yellow rhombic prisms. This base occurred in the dialysate 
(see page 265). From the non-dialyzable portion, Pouchet 
obtained another base corresponding to the formula 
C3H5N02, which he calls the “extractive matter of urine.” 
It yields precipitates with the general alkaloidal reagents, 
is non-crystallizable and is altered on exposure to air and 
resinified by hydrochloric acid. On the addition of plati- 
num chloride it is rapidly oxidized, but does not yield a 
platinochloride. The same author regards the urine as 
containing very small quantities of some pyridine bases 
which are analogous or identical with those obtained by 
Gautier and Etard from decomposing fish. 
The distinguished Italian toxicologist Selmi was, per- 
haps, the first to draw attention to the probable formation 
of basic substances in the living body during those patho- 
logical changes brought on by the presence of pathogenic chemistry of the leucomaines. 
345 
germs; and in a memoir presented to the Academy of 
Sciences of Bologna, in December, 1880, he announced 
that infectious diseases, or those in which there occurs an 
internal disarrangement of some element, either plasmic or 
histological, must be accompanied or followed by an elimi- 
nation of more or less characteristic products, which would 
be a sign of the pathological condition of the patient. To 
support this theory he examined a number of pathological 
urines, and succeeded in obtaining from them basic sub- 
stances, some of which were poisonous, others not. Thus, 
a specimen of urine from a patient with progressive paraly- 
sis gave two bases strongly resembling nicotine and coniine; 
from other pathological urines the bases obtained usually 
had either an ammoniacal or trimethylamine odor. A 
strong confirmation of Selmi’s theory is seen in the obser- 
vations made by Bouchard, Villiers, Lepine, Gau- 
tier, and others, all of whom have found basic substances 
in the urine of various diseases. 
It is now a well-established fact that the urine of disease, 
as cholera (Bouchard) and septicaemia (Feltz), etc., is far 
more poisonous than normal urine. That poisons which 
are generated within the body by the activity of bacteria 
can be excreted in the urine is seen in the fact that im- 
munity to the action of bacillus pyocyaneus has been con- 
ferred on animals by previous injection of urine taken from 
animals inoculated with that bacillus (Bouchard) or with 
filtered cultures of the same (Charrin and Buffer). 
Unfortunately, none of these bases supposedly character- 
istic of pathological urines have been isolated in a chemi- 
cally pure condition; nor has the study of normal urine 
been carried sufficiently far to show the positive absence of 
such bodies. 
Villiers has denied the existence of alkaloids in normal 
urine, and this has been confirmed experimentally by 
Stadtiiagex, who, moreover, agreed with Feltz and 
Ritter that specific organic poisons are absent from nor- 
mal urine. The observed physiological action is there- 
fore largely (70-80 per cent.), or wholly, due to the potas- 
sium salts present. 346 
BACTERIAL POISONS. 
Leucomdines of the Saliva. 
According to the statement of Gautier (1881), normal 
human saliva contains divers toxic substances in small 
quantities which differ very much in their action according 
to the time of their secretion, and probably according to 
the individual gland in which they are secreted. The 
aqueous extract of saliva at 100° is poisonous or narcotic 
in its action toward birds. To show the presence of basic 
substances, the aqueous extract was slightly acidulated with 
dilute hydrochloric acid, then precipitated by Mayer’s 
reagent; the precipitate was washed, then decomposed by 
hydrogen sulphide, and the solution filtered. The filtrate 
on evaporation gave a residue consisting of microscopic 
slender needles of a soluble hydrochloride. This salt, 
purified by extraction with absolute alcohol, forms soluble 
crystalline, but easily decomposable double salts with 
platinum chloride and with gold chloride. The solution 
of the hydrochloride produces an immediate precipitate of 
Prussian blue in a mixture of potassium ferricyanide and 
ferric chloride, and when injected into birds produces a 
condition of stupor. 
Leucomaines from other Tissues of the Body. 
Selmi’s work upon the formation of ptomaines during 
the process of putrefaction led many investigators to doubt 
the production of these bases by the decomposition of the 
proteid or other complex molecules. To substantiate this, 
a number of chemists, especially Italian, endeavored to 
show that Selmi’s bases, to a large extent at least, exist 
preformed in the various tissues. Paterno and Spica 
(1882) succeeded in extracting from fresh blood as well as 
from fresh albumin of eggs substances identical, or at least 
similar, to those designated under the name of ptomaines. 
Their observations, however, were confined to the detection 
of alkaloidal reactions in the various extracts obtained by 
Dragendorff’s method, and at no time were they in 
possession of a definite chemical individual. Marino- CHEMISTRY of the leucomaines. 
347 
Zuco (1885) was more successful, inasmuch as he succeeded 
in obtaining from fresh tissues and organs relevant quan- 
tities of a base identical with choline, and, in addition, he 
obtained extremely minute traces of other alkaloidal bodies. 
One of these, obtained by the Stas method from the liver 
and spleen of an ox, exhibited in hydrochloric acid solution 
a beautiful violet fluorescence resembling very much that 
of the salts of quinine. A similar base, probably identical 
with this one, was obtained by Bence Jones and Dupre 
(1856) from liver, nerves, tissues, and other organs, and 
was named by them “ animal chinoidine.” A greenish- 
blue fluorescence is frequently observable in the alcoholic 
extracts of decomposing glue as well as from other putrefy- 
ing substances. From a number of very thorough experi- 
ments, he concluded that basic substances do not preexist 
in fresh organs, but that the acids employed in the process 
of extraction exert a decomposing action upon the lecithin 
present in the tissues, resulting in the formation of choline. 
He further showed that the method of Dragendorfe, on 
account of the larger quantity of extractives which form, 
invariably gave a larger yield of this base than did the 
Stas-Otto method. Similar observations were made by 
Guareschi and Mosso, by Coppola and others. At the 
present time there is no doubt that some basic substances, 
among these choline, can be formed by the action of re- 
agents, and, on the other hand, it is equally well demon- 
strated that similar bases do preexist in the physiological 
condition of the tissues and fluids of the body. 
Recently R. Wurtz has obtained from normal blood a 
number of crystalline products of alkaline reaction, which 
form vvell-crystallizable double salts with gold, platinum, 
and mercuric chlorides. These, however, have not been 
as yet subjected to analysis, because of the minute quan- 
tities which were isolated. 
Morelle (1886) showed the presence in the spleen of 
the ox of a base, the hydrochloride of which crystallized 
in deliquescent needles and likewise formed crystalline 
platino- and aurochlorides. From experiments made by 
Laborde, the base would seem to possess decided toxic 348 
bacterial poisons. 
properties, bringing on a dyspnoeic condition with con- 
vulsive movements and loss of motion. The post-mortem 
examinations revealed an extended visceral cedematous 
infiltration, and stoppage of the heart in systole. 
A. W. Blyth has claimed to have isolated from milk 
two alkaloidal substances, namely galactine, the lead salt 
of which is said to have the formula Pb203C54H18N4025, 
and lactochrome, the mercury salt of which is represented 
by the formula Hg0C6H18N06. 
Leucomdines of the Venoms of Poisonous Serpents. 
The study of the chemistry of the venoms of serpents 
and of batrachians is fraught with so many difficulties and 
with so much danger, that we cannot wonder at the present 
unsatisfactory condition of our knowledge in regard to the 
poisonous principles which they contain. Much of the work 
that has been done hitherto is not only inaccurate and very 
contradictory, but is far from meeting the requirements of 
exact toxicological research. From recent investigations 
it seems, however, to be quite certain that the most active 
constituent of the venom of serpents is not alkaloidal in 
its nature as has been supposed by some. In 1881 
Gautier announced the isolation of two alkaloids from 
the venom of the cobra which gave precipitates with tannin, 
Mayer’s reagent, Nessler’s reagent, iodine in potas- 
sium iodide, etc. They formed crystallizable platinochlo- 
rides and aurochlorides, and also crystalline, neutral, some- 
what deliquescent hydrochlorides. The neutral or slightly 
acid solutions produced an immediate precipitate of Prus- 
sian blue in a mixture of potassium ferricyanide and ferric 
chloride. These substances possess a decided physiological 
action, though Gautier himself does not consider them 
to be the most dangerous constituents of the venoms. This 
observation of Gautier as to the presence of distinct basic 
substances in venoms is at variance with that of Wolcott 
Gibbs, who has been unable to obtain an alkaloid from the 
rattlesnake (Crotalus) venom. S. Weir Mitchell and 
E. T. Reichert likewise state that they have been utterly chemistry of the leucomaines. 
349 
unable to substantiate Gautier’s statements. Still more 
recently Wolfenden, in an elaborate paper on the nature 
of cobra venom, has confirmed Wolcott Gibbs as to the 
entire absence of any alkaloidal body. 
Mitchell and Reichert have made a careful study of 
the venoms of various serpents, such as cobra, rattlesnake, 
moccasin, and Indian viper, and have succeeded in isolating 
two proteid constituents, one belonging to the class of 
globulins and the other to the peptones. The peptone is 
said to be non-precipitable by alcohol. According to 
them, the globulin constituent consists of at least three 
distinct globulins. They found that boiling coagulates 
and destroys the globulin as a poison, but leaves the 
venom peptone toxically unchanged, so that the solution, 
though still poisonous, fails to produce the characteristic 
local lesions due to fresh or unboiled venom. On the other 
hand, Gautier asserts that the venom is not sensibly 
altered on being heated to 120°-125°, and that the toxic 
action remains constant even when all the proteid con- 
stituents are removed, thus showing that the toxic action 
cannot be attributed to the albuminoids. The venom pep- 
tone from the rattlesnake or the moccasin, however, when 
injected into animals produced toxic effects which were 
marked by an oedematous swelling over the site of injection; 
the tumor was filled with serum, and so also was the sub- 
cutaneous cellular tissue. Furthermore, a gradual breaking 
down of the tissues occurred, accompanied by rapid putre- 
factive changes and a more or less extensive slough. That 
peptones may possess intensely poisonous properties has been 
shown to be the case by a number of authors, among whom 
may be mentioned Schmidt-Mulhelw, Hofmeister, 
Pollitzer, and others. Brieger has, moreover, demon- 
strated that the formation of peptones in the process of 
digestion is accompanied by the development of a toxic 
ptomaine which he has named peptotoxine. 
The venom globulins, on the other hand, though present 
in less quantity than the peptones, induced the same re- 
markable local effects seen on injection of the pure venom. 350 
BACTERIAL POISONS. 
They cause local bleedings, destroy the coagulability of the 
blood, and rapidly corrode the capillaries. * 
These results of Mitchell and Reichert, which are 
given here somewhat in full, have been questioned by 
Wolfenden, who, while agreeing in the main that the 
poisonous property of venom is due to proteid constituents, 
regards their peptone not as a true peptone, but rather as 
one or more bodies of the albumose group of proteids. He 
likewise' regards the globulin of moccasin venom to be 
some other proteid body. According to him, the cobra 
venom owes its toxicity to the proteids, globulin, serum- 
albumin, acid albumin. Occasionally there seem to be 
present traces of peptone and of hemialbumose. 
Brieger was at first apparently inclined to believe that 
the action of venoms is due to animal alkaloids, on the ground 
that these bases are extremely soluble, and hence always go 
into solution along with the likewise very soluble proteid 
constituents, and that the difficulty in their isolation lies in 
the elimination of these proteids. Since then Brieger 
and Frankel pointed out the poisonous nature of some 
bacterial proteids, and also showed that cobra poison yields 
with alcohol a precipitate which gives proteid reactions. 
The proteids of serpents’ venom should be compared 
with the poisonous proteids formed by the activity of the 
pathogenic bacteria, as well as with similar compounds, 
the p/iytalbumoses of castor seeds, jequirity, etc. Fossibly 
similar compounds will be found in croton aud other 
species of ricinus, jatropha, loco-weed, etc. The poisons 
secreted by certain spiders and fish may be mentioned in 
this connection. 
Cloez and Gratiolet in 1852 examined the poison 
contained in the cutaneous pustules of some batrachians, 
and succeeded in extracting a substance which gave a white 
precipitate with mercuric chloride and formed a platinum 
double salt. Beyond this meagre information very little is 
known in regard to the character of these poisons, though 
Zalesky, in 1866, announced the isolation of an alkaloid 
to which lie assigned the formula C34H60N2O5, and which 
he named salamandarine. According to Hutartre (1890) CHEMISTRY OF THE LEUCOMAINES. 
351 
this base is a leucoma'ine. and similar products, but with 
different physiological action, are to be found in other 
batrachians, as the toad, triton (?), green and red frogs, 
and in the epidermis of some fish. According to Calmeil, 
the poison from the toad contains methyl-carbylamine and 
isocyanacetic acid. 
Table of Leucomaines. 
Formula. 
Name. 
Discoverer. 
Source. 
Physiological action. 
C6 h5 n6 
Adenine. 
Kossel. 
Nuclein-contain- 
ing organs. 
Non-poisonous; muscle 
stimulant. 
c5 h4 n4 0 
Hypoxanthine. 
Scherer. 
Nuclein-contain- 
ing organs. 
Non-poisonous; muscle 
stimulant. 
c6 h5 n5 0 
Guanine. 
Unger. 
Nuclein-contain- 
ing organs, 
guano. 
Non-poisonous; muscle 
stimulant. 
c5 h4 n4 o2 
Xanthine. 
Marcet. 
Nuclein-contain- 
ing organs, 
calculi. 
Non-poisonous; muscle 
stimulant. 
c6 h6 n4 o2 
Heteroxanthine. 
Salomon. 
Urine. 
c7 h8 n4 o2 
Paraxanthine. 
Thudichum 
Salomon. 
Poisonous. 
cT h8 n4 o3 
Oarnine. 
Weidel. 
Liebig’s meat 
extract. 
Non-poisonous; muscle 
stimulant. 
c4 h5 n5 0 
Pseudoxanthine(?) 
Gautier. 
Muscle. 
c6 H14N2 
Gerontine. 
Grandis. 
Liver of dogs. 
Poisonous. 
C2 H5 N (?) 
Spermine. 
Schreiner. 
Sperma, in tis- 
sues ofleuco- 
cythsemics. 
Non-poisonous. 
C6 H8 n4 0 
Cruso-creatinine. 
Gautier. 
Muscle. 
C5 H10N4 0 
Xantho-creatinine 
“ 
“ 
Poisonous. 
C9 H]gN7 O4 
Amphi-creatine. 
“ 
“ 
C„H24Ni0O5 
Unnamed. 
“ 
“ 
0i2H„5Nn05 
“ 
“ 
“ 
c7 h12n4 o2 
“ 
Pouchet. 
Urine. 
os h5 no. 
“ 
“ 
“ 
O5 
Salamandarine. 
Zalesky. 
Salamander. 
Poisonous. CHAPTER XIII. 
THE AUTOGENOUS DISEASES 
All living things are composed of cells. The simplest 
forms of life are unicellular, and in these all the functions 
of life devolve upon the single cell. Absorption, secretion, 
and excretion must be carried on by the same cell. A 
collection of unicellular organisms might be compared to a 
community of men with every individual his own tailor, 
shoemaker, carpenter, cook, farmer, gardener, blacksmith, 
etc. However, only the lowest forms of life are unicellular; 
all others are multicellular. In the higher animals there 
is a differentiation not only in the size aud structure of the 
cells, but in the labor which they perform. The body of 
man may be compared to a community in which labor has 
been specialized. Certain groups of cells, which we desig- 
nate by the term “ organ/’ take upon themselves the task 
of doing some special line of work, the well-doing of which 
is essential to the health, not only of that group, but of 
other groups as well, or of the body as a whole. There 
is an interdependence among the various organs. Certain 
groups of cells supply the fluids or juices which act as 
digestants, and among these there is again a division of 
labor. The salivary glands supply a fluid which partially 
digests the starch of our food ; the peptic glands supply 
the gastric juice which does the preliminary work in the 
digestion of the proteids; while the pancreatic juice com- 
pletes the digestion of the starches begun in the mouth, of 
the proteids begun in the stomach, and does the special 
work of emulsifying the fats. But even some of these 
products of complete digestion would be harmful should 
they enter the circulation unchanged. The peptones must 
be converted into serum-albumin by the absorbing mechan- 
ism of the walls of the intestines, aud while 10 per cent. THE AUTOGENOUS DISEASES. 
853 
of the fat of the food is split up into glycerin and fatty 
acids by the action of the pancreatic juice, a much smaller 
per cent, enters the thoracic duct in this divided form. 
The food may be taken in proper quality and quantity; 
the digestive juices may do their work promptly and 
properly, but if the absorbents fail to perform their func- 
tions properly, disease results. It may happen that the 
failure lies in improper or imperfect assimilation and the 
result becomes equally disastrous, and with the effects of 
non-elimination we are fairly conversant. Of the myriads 
of cells in the healthy human body there are none which 
are superfluous. It is true that among these ultimate 
entities of existence, death is constantly occurring, but in 
health regeneration goes on with equal rapidity and each 
organ continues to do its.daily and hourly task. The 
microscope has made us familiar with the size and shape 
of the various cells of the body, and students of pathology 
have described the alterations in form and size character- 
istic of various disease states. But we must remember 
that in the study of these ultimate elements of life there 
are other things, besides their morphological history, to 
investigate. They are endowed with life, and they, as well 
as the germs, have a physiology and chemistry which we 
know but slightly. They are influenced beneficially or 
harmfully, as the case may be, by their environment. 
They grow and perform their functions properly when 
supplied with the needed pabulum. They are not immune 
to poisonous agents. They are injured when the products 
of their own activity accumulate about them. 
The object in writing this chapter has been to collect 
what evidence we may concerning those diseases which 
arise from imperfect or improper activity of the cells of the 
body, not due to the introduction of foreign cells. To 
designate this class of diseases we have selected the word 
autogenous, and we understand that in these diseases the 
materies morbi is a product of some cell of the body, and 
not, as in the case of the infectious diseases, of cells intro- 
duced from without the body. 
It is true, without exception so far as we know, that the 354 
BACTERIAL POISONS. 
excretions of all living things, plants and animals, contain 
substances which are poisonous to the organisms which 
excrete them. A man may drink only chemically pure 
water, eat only that food which is free from all adultera- 
tions, and breathe nothing but the purest air, free from all 
organic matter, both living and dead, and yet that man’s 
excretions would contain poisons. Where do these poisons 
originate? They are formed within the body. They 
originate in the metabolic changes by which the complex 
organic molecule is split up into simpler compounds. We 
may suppose—indeed, we have good reasons for believing— 
that the proteid molecule has certain lines of cleavage along 
which it breaks when certain forces are applied, and that 
the resulting fragments have also lines of cleavage along 
which they break under certain influences, and so on until 
the end-products, urea, ammonia, water, and carbon-dioxide 
are reached ; also that some of these intermediate products 
are highly poisonous has been abundantly demonstrated. 
The fact that the hydrocyanic acid molecule is a frequent 
constituent of the leucoma'ines is one of great significance. 
We know that chemical composition is an indication of 
physiological action, and the intensely poisonous character 
of some of the leucoma'ines conforms to this fact. It 
matters not whether the proteid molecule be broken up by 
organized ferments, bacteria, or by the unorganized fer- 
ments of the digestive juices, by the cells of the liver or by 
those still unknown agencies, which induce metabolic 
changes in all the tissues—in all cases poisons may be 
formed. These poisons will differ in quality and quantity 
according to the proteid which is acted upon, and according 
to the force which acts. 
Peptones formed during digestion do not in health reach 
the general circulation. When injected directly into the 
blood they act as powerful poisons. They destroy the 
coagulability of the blood, lower blood-pressure, and in 
large quantities cause speedy death. Brunton attributes 
the lassitude, depression, sense of weight in the limbs, and 
dulness in the head occurring in the well-fed, inactive man, 
after his meals, to poisoning with peptones. The remedy THE autogenous diseases. 
355 
which he proposes is less food, especially less nitrogenous 
food, and more exercise. That some substance resulting 
from the proteids of the food is the cause of this trouble 
Brunton thinks is evidenced by the fact that the weak- 
ness and languor are apparently less after meals consisting 
of farinaceous foods only. 
That peptone finds its way into the general circulation 
frequently is shown by its detection in the urine in many 
diseased conditions, some of which are infectious and others 
autogenous in character. However, propeptonuria, or albu- 
mosuria, is more common than peptonuria, and we have 
already seen that many of the bacterial albumoses are among 
the most highly poisonous bodies known, but the action of 
the albumoses formed during digestion has not, so far as 
we know, been studied. The valuable work of Kuhne and 
Chittenden on the chemical character of these bodies 
should be supplemented by a thorough investigation of their 
physiological effects when injected into the blood. It is more 
than probable that valuable information would be secured 
by such studies. That albumose is frequently found in the 
urine is shown by the following list of diseases in which 
it has been observed, given in the last edition of the work 
of Neubauer and Vogel on the urine: Kosner has 
found it in spermatorrhoea; Koppen, in mental diseases 
without spermatorrhoea ; Kahler, in osteomalacia; Bence 
Jones, in multiple myeloma; Senator and others, in 
dermatitis, intestinal ulcer, liver abscess, croupous pneu- 
monia, apoplexy, vitium cordis, resectio coxse, parame- 
tritis, endocarditis, typhoid fever, nephritis, phthisis, etc.; 
Loeb, in measles and scarlet fever; Leube, in urticaria; 
and Lassar, after inunctions of petroleum. Kottnitz, 
Furstner, and others, find albumose frequently in the 
urine in mental diseases. Evidently, there is much to 
learn from the study of the conditions accompanied by the 
elimination of the albumoses in the urine. It is more than 
probable that the acute Bright’s disease following scarlet 
fever, diphtheria, and the other acute infectious diseases, 
owes its existence to the poisonous albumoses of these dis- 
eases. Prior has recently shown that undigested egg 356 
BACTERIAL POISONS. 
albumin is sometimes absorbed and produces marked dis- 
turbances. A boy, after eating sixteen raw eggs, had a 
high fever accompanied by the appearance of both albumin 
and hemoglobin in the urine. 
Brieger obtained by digesting fibrin with gastric juice 
a substance which gives reactions with many of the general 
alkaloidal reagents and to which he has given the name 
“ peptotoxine.” A few drops of a dilute aqueous solution 
of this substance sufficed to kill frogs within fifteen min- 
utes. The frogs became apparently paralyzed and did not 
respond to stimuli. Slight tremor was perceptible in the 
muscles of the extremities. Rabbits of about one kilo- 
gramme weight were given from 0.5 to 1 gramme of the 
extract subcutaneously. About fifteen minutes after the 
injection, paralysis beginning in the posterior extremities 
set in; the animal fell into a somnolent condition, sank 
and died. In some rabbits several hours elapsed before 
the above-mentioned symptoms appeared. 
Peptotoxine was found by Brieger. to be formed not 
only by the digestive juice, but to be among the first 
putrefactive products of proteids, as fibrin, casein, brain 
substance, liver, and muscle. 
It is highly probable that many of the nervous symptoms 
which accompany some forms of dyspepsia are due to the 
formation and absorption of poisonous substances. 
In some persons the tendency to the formation of poisons 
out of certain foods is very marked. Thus, there are some 
to whom the smallest bit of egg is highly poisonous; with 
others, milk will not agree; and instances of this kind are 
sufficiently numerous to give rise to the adage, “ What is 
one man’s meat is another man’s poison.” 
Brunton is of the opinion that the condition which we 
term “ biliousness,” and which is most likely to exist in 
those who eat largely of proteids, is due to the formation 
of poisonous alkaloids; but of this we have no positive 
proof. 
Whether or not the unorganized digestive ferments ever 
find their way into the blood in quantity sufficient to cause 
deviations from health, we are not in a position to state THE AUTOGENOUS DISEASES. 
357 
definitely. The older physiological chemists teach us that 
pepsin and trypsin are frequent, if not constant constituents 
of normal urine, but their experiments were made without 
any reference to the possibility of the ferments which they 
found being formed by the bacteria of the urine, and after 
carefully going over the literature of the subject we are not 
prepared to pass judgment on the truth of their statements. 
However this may be, the fact that these ferments manifest 
a marked toxicological effect when introduced into the 
blood is of great interest, especially at this time. Hilde- 
beandt has recently reported the results of some experi- 
ments made by himself upon this subject. He finds that 
a fatal dose of pepsin for dogs is from 0.1 to 0.2 gramme 
per kilogramme of body weight. The subcutaneous injec- 
tion of these quantities is followed by a marked elevation 
of temperature, which he designates as “ ferment fever.” 
This fever begins within an hour after the injection, 
reaches its maximum after from four to six hours, and 
may continue for some days. On the day preceding death, 
the temperature generally falls below the normal. During 
the period of elevation there are frequent chills. 
The symptoms which accompanying the fever vary 
somewhat with the species of animal. Rabbits lose flesh 
nothwithstanding the fact that they continue for a while 
to eat well, they become very weak, and death is preceded 
by convulsive movements. Dogs tremble in the limbs, be- 
come uncertain in gait, and vomiting, dyspnoea and coma 
are followed by death. 
On section there is observed parenchymatous degenera- 
tion of the muscles of the heart and similar changes in the 
liver and kidney. There are abundant hemorrhages in the 
intestinal canal, in Peyee’s patches, in the mesenteric 
glands; and in the lungs in cats. Thrombi are frequently 
found in the lungs and in some cases in the kidneys. 
The effect upon the coagulability of the blood is worthy 
of note. At first there is a period during which the coagu- 
lability of the blood is greatly lessened, then follows a 
period of greater rapidity in coagulating, and it is in this 
latter stage that the thrombi are formed. 358 
BACTERIAL POISONS. 
These experiments are interesting not only as a possible 
explanation of the cause of some of the autogenous fevers, 
which will be discussed later, but in view of the present 
tendency to inject such complex animal solutions as 
Brown- S£quard’s elixir and Koch’s lymph subcutane- 
ously, and they will probably cause us to exercise a little 
more care in this direction. 
That certain febrile conditions are autogenous there can 
be no doubt. These, like other diseases originating within 
the system, may be due to either of the following causes: 
1. There may be an excessive formation of poisonous sub- 
stances in the body. Thus, Bouchard has shown that 
the urine excreted during the hours of activity is much 
more poisonous than that excreted during the hours of rest. 
Both physical and mental labor are accompanied by the 
formation of these deleterious bodies, and if the hours of 
labor are prolonged and those of rest shortened, there will 
be an accumulation of effete matters within the system. 
2. The accumulation of the poisonous matters may be due 
to deficient elimination. 3. Some organ whose duty it is 
to change harmful into harmless bodies may fail to prop- 
erly perform its functions. Illustrations of diseased con- 
ditions arising from these several causes will be given. 
First, we may mention fatigue fever, which is by no 
means uncommon, and from which the overworked physi- 
cian not infrequently suffers. One works night and day 
for some time; elimination seems to proceed normally; 
but after a few days there is an elevation of temperature 
of from one to three degrees, the appetite is impaired, and 
then if the opportunity for rest is at hand sound and rest- 
ful sleep is impossible. The tired man retires to his bed 
expecting to fall asleep immediately, but he tosses from side 
to side all night, or his sleep is fitful and uurefreshing. 
The brain is excited and refuses to be at rest. The senses 
are alert, and all efforts to sink them in repose are unavail- 
ing. Fatigue fever is frequently observed in armies upon 
forced marches, especially if the troops are young and un- 
accustomed to service. Mosso has studied this fever in THE AUTOGENOUS DISEASES. 
359 
the Italian army. He states that in fatigue the blood is 
subjected to a process of decomposition brought about by 
the infiltration into it from the tissues of poisonous sub- 
stances, which, when injected into the circulation of healthy 
animals, induce malaise and all the signs of excessive ex- 
haustion. It is possible that in this decomposition of the 
blood the fibrin-ferment, which, according to Schmidt, is 
held in combination in the colorless corpuscles, is liberated; 
and it has been shown by Edelberg that the injection of 
small quantities of free fibrin-ferment into the blood causes 
fever, while the injection of larger quantities is followed by 
the formation of thrombi, as has been demonstrated by the 
experiments of Edelberg, Bonne, Birk, and Kohlar. 
Fatigue fever is often accompanied, especially during the 
period of elevation, by chilly seusations, and consequently 
it is pronounced malarial and quinine is administered, but 
it does no good—often harm, by increasing cerebral excite- 
ment. The proper treatment is prolonged rest, with proper 
attention to elimination. 
Then there is the fever of exhaustion, which differs from 
fatigue fever only in degree. It is brought on by pro- 
longed exertion without sufficient rest and often without 
sufficient food. The healthy balance between the formation 
and elimination of effete matter is disturbed, and it may 
be weeks before it is reestablished—indeed, it may never 
be regained, for some of those cases terminate fatally. The 
fever of exhaustion may take on the typhus form, delirium 
may appear, muscular control of the bowels may be lost, 
and death may result. 
That the fever of exhaustion has been mistaken for 
typhoid by some of the ablest clinical teachers is shown by 
Peter in the following quotation. “It was in 1852,” says 
he, “ when entering upon my clinical studies and ardent in 
my attendance at the clinic of Chomel, I was witness of 
the following instance: A young man was received under 
the celebrated professor’s charge suffering from prostration, 
muscular pain, and rhachialgia. Chomel made the exam- 
ination with all the care and attention used by him ; then 360 
BACTERIAL poisons. 
—as was also usual with him in the presence of the patient 
—he gave the diagnosis in Latin, which was 1 Aut febris 
peyerica, aut variola incipientis ’ (either typhoid fever or 
incipient smallpox). I felt rather dissatisfied at a diagnosis 
so little precise by one so eminent in his art. The truth of 
the matter was, though Chomel was not aware of it, this 
young fellow in a state of destitution had walked from 
Compiegne to Paris, sleeping by the wayside at night and 
nourishing himself with such refuse food as chance supplied. 
It was under such circumstances the patient had developed 
febrile symptoms. The day after his admission, and simply 
from rest in bed, he felt better, and the day following he 
was altogether well.” 
That all cases of the fever of exhaustion do not terminate 
so rapidly as that instanced above many physicians know. 
We have seen at least one such case terminate fatally. 
Then, again, there is the fever of non-elimination, which 
all physicians of experience have observed. There is a 
feeling of languor, the head aches, the tongue is coated, the 
breath offensive, and the bowels constipated. The physi- 
cian fears typhoid fever, but finds that a good, brisk cathar- 
tic dissipates all unpleasant symptoms, and the temperature 
falls to the normal. This fever is also liable to appear 
among those who are confined to bed from other causes. 
Brunton says : “ No one who has watched cases of acute 
diseases, such as pneumonia, can have failed to see how a 
rise of temperature sometimes coincides with the occurrence 
of constipation, and is removed by opening the bowels.” 
The surgeon and obstetrician have often had cause to rejoice 
when they have found a fever, which they feared indicated 
septicaemia, disappearing after free purgation. 
Bouchard has shown that normal feces contain a highly 
poisonous substance, which may be separated from them by 
dialysis, and which, when administered to rabbits, produces 
violent convulsions. He estimates that the amount of 
poisonous alkaloids formed in the intestines of a healthy 
man each twenty-four hours would be quite sufficient to 
kill, if it was all absorbed. He proposes the term “ ster- THE AUTOGENOUS DISEASES. 
361 
corsemia” for that condition which results from arrest of 
excretion from the intestine. 
It is more than probable that the poisons of the intes- 
tines are due to the bacteria which are normally present; 
but this would not exclude the fever of non-elimination 
from the list of autogenous diseases. The bacterial cells 
which are normally present in the intestines cannot be 
regarded as invaders from without. 
It would seem from some recent studies that not all sur- 
gical fevers are due to bacterial activity. The absorption 
of aseptic blood-clots and of disintegrated tissue in cases of 
complicated fractures and contusions of the joints is accom- 
panied by an elevation of the temperature above normal. 
A like result may follow the intravenous injection of a 
sterile solution of haemoglobin or of the blood of another 
animal. The causative agent in the production of these 
fevers remains unknown. In the blood of twelve out of 
fifteen patients with aseptic fever, at the clinic of Noth- 
nagel, Hammerschlag has found free fibrin-ferment, 
but in five persons without fever he found the same sub- 
stance in the blood. This leaves the causative agent in the 
production of the aseptic, or, more properly speaking, the 
non-bacterial, fevers unknown. 
The chemical theory of so-called uraemia has received 
support in recent researches, notwithstanding the fact that 
the old idea that urea is the active poison and the theory 
of Frereiches that ammonium carbonate is the active 
agent have been abandoned. 
Landois laid bare the surface of the brain in dogs 
and rabbits, and sprinkled the motor area with creatine, 
creatinine, and other constituents of the urine. Urea, 
ammonium carbonate, sodium chloride, and potassium 
chloride had but slight effect; but creatine, creatinine, 
and acid sodium phosphate caused clonic convulsions on 
the opposite side of the body which later became bilateral. 
The convulsions continued at intervals for from two to 
three days, when, growing gradually weaker, they disap- 
peared. Landois concludes that chorea gravidarum is a 362 
BACTERIAL POISONS. 
forerunner of eclampsia. These experiments have been 
confirmed by Leubuscher and Zeichen. 
Falck injected into both sound and nephrotomized ani- 
mals fresh urine, urine and the ferment of Musculus and 
Lea, and urine which had undergone spontaneous decom- 
position, without producing any symptoms which were 
comparable with those observed in uraemia. However, he 
did find that if a few drops of an infusion of putrid flesh 
were added to the urine before injection all the typical 
symptoms of uraemia were induced. That the infusion of 
putrid flesh alone had no effect was also demonstrated. 
This would lead us to believe that some ferment in the 
infusion converts some constituent of the urine into a 
highly poisonous body. In this connection attention may 
be called to the fact that creatine may be converted by the 
action of certain germs into methyl-guanidine, which pro- 
duces convulsions. Whether such conversion occurs in 
uraemia or not, and if it does what the cause of it is, are 
questions which must be left for future investigations to 
decide. It would be well for someone to test the brain 
and blood of a person who has died in uraemic convulsions 
for methyl-guanidine. 
That there is a marked disturbance of tissue metabolism 
caused by the inhalation of vitiated air has been shown by 
Araki. In the urine of animals rendered unconscious by 
being kept in a confined space this experimenter found 
albumin, sugar, and lactic acid. If the animals had been 
kept without food for some days before being subjected to 
this experiment albumin and lactic acid were found, but 
no sugar appeared. This was undoubtedly due to the fact 
that the glycogen of the body had been exhausted by the 
fasting. Identical results were observed in animals which 
were poisoned with carbon monoxide. Hogs which were 
poisoned with curare, and in which the respiratory move- 
ments were maintained artificially, secreted very little 
urine; but the blood was found to contain considerable 
quantities of sugar and lactic acid. The urine of frogs in 
which the respiration was retarded by the production of 
tetanus with strychnine secreted urine containing sugar and 1 ? 
I.—Tabular View of the Reactions of Certain Ptomaines. 
Trimethylamine 
Hydrochloride, 
NiCHals.HCl 
Diethylamine 
Hydrochloride, 
NCCsHsV-H.HCl 
A Base (p- 202), 
CioHuN.HCl 
Ethylidenediamine 
Hydrochloride, 
C2H8No,2HC1 
— 
CiHi-Nu- 
Oadaverine, 
CsHhNo 
Putrescine 
Hydrochloride, 
C4Hi..N..,2HC1 
Oadaverine 
Hydrochloride, 
05Hi4N->,2HC1 
Neuridine 
Hydrochloride, 
CoH14N-.,2HC1 
——■ 
Methyl-guanidine 
Hydrochloride, 
02H7K3.HC1 
Neurine Chloride, 
CsHioN.Cl 
Choline Chloride, 
CsHhNO.CI 
Betaine Chloride, 
C5H1-.NO2.Cl 
Typhotoxine 
Hydrochloride, 
C7H17NO2.HCI 
A Base (262), 
C7H17NO2.HCI 
Base from Cul- 
tures of 
Typhoid Bacilli) 
(Brieger, ii. 69) 
Phosphotungstic 
Acid. 
White crystalline 
precipitate, easily 
soluble in water. 
Yellowish-white 
\\ 
ite precipitate, 
ible in excess. 
White precipitate, 
easily soluble in 
excess. 
White precipitate. 
White precipitate, 
easily soluble in 
excess. 
White amorphous 
precipitate, soluble 
in excess. 
White precipitate, 
insoluble in water; 
on standing be- 
comes crystallized. 
White precipitate, 
soluble in excess. 
White crystalline 
precipitate. 
White precipitate. 
precipitate. 
s< 
Phosphomolybdic 
Acid. 
Phosphoantimonic 
Acid. 
White granular 
precipitate. 
Yellow precipitate. 
Precipitate easily 
soluble in excess. 
Heavy yellow pre- 
cipitate, difficultly 
soluble in NH4OH; 
no blue color. 
White precipitate. 
Yellowish-white pre- 
cipitate, soluble in 
excess. 
Yt 
ow precipitate. 
White crystalline 
precipitate, soluble 
in excess. 
White crystalline 
precipitate. 
Yellow precipitate. 
White crystalline 
precipitate ; yel- 
low precipitate 
(Bocklisch). 
White precipitate, 
soluble in excess. 
White crystalline 
precipitate. 
White flocculent 
precipitate. 
Yellow crystalline 
precipitate. 
White crystalline 
precipitate, insolu- 
ble in excess. 
White voluminous 
precipitate. 
Voluminous pre- 
cipitate. 
White curdy pre- 
cipitate. 
Yellow precipitate. 
A precipitate, 
readily soluble in 
excess. 
Yellow crystalline 
precipitate. 
Yellow precipitate. 
White precipitate 
Yellow precipitate 
crystallizing in 
needles. 
First amorphous, 
then assumes 
cauliflower shape. 
llow needles. 
Well-formed, diffi- 
cultly soluble 
broad needles. 
Difficultly soluble 
precipitate. 
Picric Acid. 
Yellow needles. 
Yellow needles. 
Precipitates slowly 
in beautiful yellow 
needles. 
Yellow needles. 
D 
y-white preci 
pitate. 
Dirty-white, volu 
minous precipitate. 
Tannic Acid. 
Whitish precipi- 
tate. 
White amorphous 
precipitate. 
0 
amorphous preci- 
pitate. 
Potassium Bismuth 
Iodide. 
Red precipitate. 
Brick-red precipi- 
tate. 
Precipitate of red 
plates. 
C 
y precipitate, 
oon becomes 
crystalline. 
Brown precipitate. 
First amorphous, 
then crystallizes 
in needles. 
Red needles. 
Red amorphous 
precipitate. 
Brick red precipi- 
tate. 
Red amorphous 
precipitate. 
Red amorphous 
precipitate. 
Brick-red precipi- 
tate. 
Resinous precipi- 
tate. 
Brick-red amorph- 
ous precipitate. 
Reddish-brown 
precipitate. 
Potassium Mercuric 
Iodide. 
Potassium Cadmium 
Iodide. 
Yellowish precipi- 
tate. 
Oily precipitate, 
soon becomes 
granular. 
Whitish precipi- 
tate. 
Id. 
Resinous precipi- 
tate. 
Id. 
0 
Yellowish-white, 
voluminous preci- 
pitate. 
White precipitate. 
Yellowish crystal- 
line precipitate. 
Clear yellow, oily 
precipitate, soluble 
in excess; when 
sides are rubbed 
= yellow needles. 
Oily drops which 
do not become 
crystalline. 
Id. 
At first an oily pre- 
cipitate, which 
soon solidifies to 
needles. 
Id. 
Id. 
precipitate. 
crystalline pre- 
cipitate. 
Yellow crystalline 
precipitate. 
Crystalline pre- 
cipitate. 
Yellow precipi- 
tate, soluble in 
hot water. 
Gold Chloride. 
A precipitate. 
Platinum Chloride. 
Heavy yellow pre- 
cipitate. 
A precipitate. 
Precipitate in con- 
centrated solution. 
Alcoholic HgClo 
- 0. 
White granular 
precipitate, ap- 
pears slowly. 
A precipitate of 
short prisms, rather 
easily soluble. 
Mercuric Chloride. 
White precipitate. 
White granular 
precipitate. 
Alcoholic HgClo 
gives precipitate 
in cone, solution. 
Iodine in Potassium 
Iodide. 
Brown precipitate, 
solidifying after 
some time in 
plates. 
Brown oily preci- 
pitate. 
Brown precipitate. 
Brown crystalline 
precipitate. 
Brown needles. 
Oily drops 
Brown amorphous 
precipitate. 
Brown granular 
precipitate. 
Oily precipitate. 
Id 
Id. 
Brown precipitate 
precipitate. 
Id. 
Iodine in Hydriodic 
Acid. 
Id. 
Id. 
Id. 
Id. 
Id. 
Id. 
Id. 
A precipitate of 
fine needles. 
Crystalline pre- 
cipitate. 
Id. 
Id. 
Ferric Chloride 
and Potassium 
Ferricyanide. 
When perfectly 
pure, no blue 
color. 
0 
After a time gives 
a blue coloration. 
0 
blue precipitate. 
pure. 
Potassium Bichro- 
mate and Con- 
centrated Sul- 
phuric Acid. 
Fhohde’s re- 
agent = 0. 
mate = yellow, 
hardly crystalline 
precipitate. 
precipitate which 
soon disappears. 
= needles. 
To face page 362. Table II.—Ptoma s in Toxicological Examinations. 
Note.—The greater part of this Table Hass been taken direct from Grabner’s Inaugural Dissertation. 
Table III.—Reactions of Selmi’s Ptomaines. 
h 
Mydaleine 
Peptotoxine. 
Hoffa’s Ptomaine 
of Anthrax Bacil- 
Ptomaine from 
Cultures of 
Staphylococcus 
pyogenes aureus. 
Susotoxine 
Hydrochloride. 
Rorsch and 
Fassbender. 
Schwanert. 
Liehermann. 
Zuelzer and 
Sonnenschein. 
\| 
Gtelder. 
Brouardel and Boutmy. 
Baumurt 
(Liehermanu). 
Otto. 
Solvent. 
Ether 
acid). 
a. 
Ether 
(alkaline). 
6. 
Chloroform 
(alkaline). 
c. 
Amyl Alcohol 
(alkaline). 
d. 
Hydrochloride. 
Ins. 
Substance examined 
Liver of decompos- 
ing cadaver and of 
fresh liver of ox. 
Decomposing liver, 
kidney, and 
stomach. 
Putrefying stomach 
and contents. 
Muscle macerated in 
water. 
Liver, 
ach, ui 
an exl; 
conti 
kidney, stom- 
id intestine of 
umed arsenic- 
ining body. 
Cadaver. 
Death by asphyxia. 
Cadaver. 
Death by hydro- 
cyanic acid. 
Parts of putrefying 
goose and cadaver; 
In water 18 months. 
White precipitate, 
soluble in excess. 
Voluminous white 
precipitate. 
White precipitate. 
White precipitate 
soluble in excess. 
Heavy white 
precipitate 
soluble in NH4OH 
Iodine in Hydriodic Acid 
Id. 
Precipitates in 
two crystalline 
forms. 
Crystalline 
precipitate. 
Reddish-brown 
precipitate with 
deliquescing 
crystals. 
Solvent 
Residue 
Ether (acid and alka- 
line). 
Amorphous, not 
bitter. 
Ether (alkaline). 
Liquid, volatile; re- 
pulsive taste. 
Ether (acid and alka- 
line). 
Resinous, brownish, 
soluble in water; 
acid taste. 
Ether (alkaline). 
Greasy brownish 
mass, with crystals. 
Ether (acid). 
Yellow, amorphous; 
Petroleum ether 
Gold Chloride .... 
Id. 
Yellow amorphous 
precipitate 
Id. 
Heavy yellow pre- 
cipitate. 
Yellow flocculent 
precipitate. 
Yellowish precipi- 
tate soluble in 
Bio 
vn extract. 
Alkaline. 
Alkaline, volatile 
(alkaline). 
Bright yellow oil. 
A precipitate. 
Yellowish pre- 
cipitate. 
NH4OH; no blue 
color. 
liquid; odor that of 
urine of mice. 
taste sharp, 
bitter. 
Mercuric Chloride . . 
Phosphomolybdic Acid . 
Ill 
White precipi- 
tate. 
Violet or dark 
blue. 
Whitish preci- 
pitate. 
precipitate. 
Tannic Acid. 
Gold Chloride 
White precipitate. 
Gradual cloudiness. 
Bluish yellow pre- 
cipitate. 
Dirty-yellow six- 
sided stars. 
White precipitate. 
White precipitate. 
Yellow crystalline 
precipitate 
Brownish-yellow 
precipitate. 
White precipitate. 
White precipitate. 
White precipitate. 
Violet precipitate. 
Same as colchicine; 
soluble in alcohol. 
Same as colchicine. 
Yellow oily preci- 
pitate. 
Clear yellow pre- 
cipitate. 
Yellow needles. 
O 
A precipitate. 
Concentrated Sulphuric 
Acid (warmed). 
Violet red. 
Violet or yellow- 
ish-brown. 
Reddish 
coloration. 
Sulphuric Acid and Potas- 
sium Dichromate. 
Gradually passes 
into green. 
White granular 
precipitate. 
Flocculent yellow- 
ish-white precipi- 
Platinum Chloride 
Yellowish precipi- 
tate. 
Id. 
A precipitate, 
Id. 
colchicine 
= 0. 
Iodic Acid 
Reduction. 
Reduction. 
Reduction. 
Reduction. 
tilt 6. 
Iodine in Potassium 
Yellowish-brown 
precipitate. 
Clear brown precipi- 
Yellow to dark 
brown. 
Kermes-brown pre- 
cipitate. 
fd. 
Kermes-brown precipitate. 
Kermes-brown preci- 
pitate. 
Same as colchicine. 
Iodic Acid + Sulphuric 
Acid -f- Soda. 
Nitric Acid 
Violet. 
Dirty-brown oily 
precipitate. 
needles. 
Frohde’s Reagent 
Splendid blue, later 
green. 
Orange-red. 
Slight yellow. 
Yellow oily preci- 
pitate. 
A compact yellow 
precipitate. 
Golden yellow 
precipitate. 
warming 
stronger). 
Phosphomoi.yhdic 
Acid 
Yellow precipitate, 
on warming green ; 
with NH4OH, blue. 
Yellow precipitate, 
with NH4OH, blue. 
Yellow precipitate. 
Heavy flocculent 
precipitate. 
Yellow precipitate. 
White precipitate. 
blue with NH4OH. 
Hydrochloric Acid -f Sul- 
phuric Acid. 
Slight violet 
Violet color with 
odor of haw- 
Id. 
Faint rose color. 
Sulphuric Acid 
Dirty brownish-yel- 
low, unchanged on 
warming. 
Reddish-brown, then 
grass-green. 
Yellow. 
Colorless, then slight 
reddish-violet color. 
On warming, violet. 
In the cold, brown- 
ish violet. 
Colorless; on warm- 
Yellow color. 
yellow. 
ing, violet. 
Sulphuric Acid+Bromine 
Frohde’s Reagent . . . 
Red (permanent) 
Yellowish-brown 
to violet or yel- 
low. 
Cloudiness. 
Slight violet, 
more distinct 
on standing. 
iodide = a red 
precipitate. 
Sulphuric Acid and 
No odor of butyric 
phuric acid. 
Red. 
Oily drops. 
A precipitate. 
acid. 
Addition of alcohol 
produces a slight 
precipitate. 
Id. 
MATE 
Nitric Acid 
Yellow spots on eva- 
poration. 
Id. 
Dark-yellow color, 
Ferric Chloride .... 
Hydrochloric Acid . . . 
Rose-colored 
precipitate. 
Microscopic 
needles united in 
tufts. 
0 
Yellowish precipi- 
tate. 
concentrate! 
= carinin 
which with 
IINO:1 
red, 
water 
A precipitate. 
Yellowish-white 
Id. 
gives yellow. 
A precipitate. 
precipitate. 
Hydrochloric Acid 
Deliquescent white 
needles. 
Cherry-red on heat- 
Colorless 
ing. 
Phosphoric Acid (diluted) 
On warming, 
violet color. 
Mercuric Chloride 
White crystalline 
precipitate. 
Dirty-white precipi- 
tate. 
White cloudiness. 
Curdy white precipi- 
tate. 
White precipitate. 
Dirty-brown oily 
precipitate. 
Brown precipitate. 
Brownish-yellow 
precipitate. 
Dirty-brown 
precipitate. 
Sulphuric Acid (diluted) 
Violet pass- 
ing into yel- 
low; odor of 
hawthorn. 
Potassium Mercuric 
Chloride 
Id 
Abundant precipi- 
tate. 
Same as colchicine. 
becomes black. 
Id. 
Id. 
Oily drops. 
Chlorine Water 
Heavy white precipi- 
tate. 
Id. 
A precipitate, 
Potassium Platjnio Cya- 
nide. 
colchicine 
= 0. 
mmediate intense 
blue color. 
Berlin blue. 
Intense blue color 
Picric Acid 
Abundant yellow 
precipitate Ionic 
acid is reduced. Sil- 
ver nitrate, white 
precipitate with re- 
Potassium ferri- 
Zeisel’s reaction=0, 
Potassium Argentic Cya- 
nide. 
Platinum Chloride . . 
cyanide is reduced. 
II0SO4 + BaH.,02 = 
brick-red ; on warm- 
ing, violet. 
colchicine, green. 
Ferric salts = blue. 
Millon’s reagent 
showed presence of 
f Occasionally' 
( precipitates. 
t 
1 
duction ofsiver. Fer- 
ric chloride = 0. 
peptone. 
Potassium Dichromatf. . 
Physiological 
Action. 
When fed to pigeons 
no effect. 
Causes mydriasis and 
increase in the rate 
of heart-beat. 
In frogs, produces 
slowing of heart, 
paralysis, death. 
Non-poisonous. 
Non-poisonous. 
Non-poisonous 
Intensely poisonous. THE AUTOGENOUS DISEASES. 
363 
lactic acid. In the urine of three epileptics there were 
found albumin and lactic acid directly after the seizure. 
The factor common to all these cases is diminished oxygen- 
ation of the blood, and to this is ascribed the appearance of 
the abnormal constituents of the urine. These investiera- 
tions demonstrate the influence of impure air upon the 
chemistry of the living cells of the animal body. CHAPTER XIV. 
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Widenmann. Zeitschrift f. Hygiene, 5, 522. 
Wyssokowitsch. Zeitschrift f. Hygiene, 1, 1. INDEX. 
Adenine, 283 
Adenine-hypoxanthine, 297 
Aerobic bacteria, 17 
Agaricine, 237 
Albumins, poisonous, 61 
Albumoses, 35 
immunity from, 150 
in urine, 355 
Alcohol, basic substances in, 158 
dialysis into, 172 
effect on bacterial proteids, 173 
Alcoholic fermentation, bases in, 222 
Aldehyde collidine, 196 
Alkaloids, interference in reactions 
of, by ptomaines, 183 
separation from ptomaines, 186 
Alkapton, 281 
Amanitine, 237 
American swine-plague, 142 
Amido-valerianie acid, 231 
Amphi-creatine, 338 
Amylamine, 193 
Amylic alcohol, impurities in, 158 
Anaerobic bacteria, 17 
Alkaloids, 15 
Animal cliinoidine, 26, 243, 347 
coniine, 214 
Anthracin, 102, 104, 277 
Anthrax, 101 
theories of, 85 et seq. 
bacillus, products of, 101 et seq. 
proteid, 103, 171 
albumose, 103, 156, 171 
Apricots, poisonous, 52 
Arginine, 189, 242, 333 
Aromine, 344 
Aselline, 229 
Aseptic fever, 361 
Asiatic cholera, 104 
bacillus of, products of, 107 
Atropine-like substances, 27, 179 
Autogenous diseases, 14, 352 
Azulmic acid, 283, 288, 337 
i 1}ACILLUS butyricus, 16 
I iJ enteriditis, 50 
j Bacon, poisonous, 51 
| Bacteria, classification of, 13 
in summer diarrhosas, 133, 
136 
| Bacterial cellular proteids, 19 
method of extraction, 
130 
poisons, definition and classifica- 
tion of, 15 
historical sketch of, 22 
foods containing, 36 
relation to infectious dis- 
eases, 84 
proteids, 18 
method of extraction, 170, 
171 
Bacterium allii, 203 
Batrachians, poison of, 350 
I Beer, colchicine-like substance in, 
183 
Benzol, impurities in, 158 
Bergmann and Schmiedeberg’s me- 
thod, 169 
Betaine, 248, 334, 343 
Bibliography of the leucomaines, 
381 
! of the ptomaines, 364 
Bilineurine, 237 
Biliousness, 356 
Blood, germicidal properties, 153 
leucomaines in, 347 
Bocklisch’s base, unnamed, 272 
Botulinic acid, 46 
Bread, poisonons, 83 
Brieger’s bases, unnamed, 195, 255, 
261, 271, 272, 273, 274, 275 
methods, 161 
disadvantages of, 170 
Brouardel’s veratrine, 204 
Bujwid’s cholera-reaction, 110 
Butylamine, 193 388 
INDEX 
Cadaveric coniine, 176,214 
Cadaverine, 34, 107, 212 
Caffeine, 316 
Canned meats, poisonous, 52 
Caproylamine. 194 
Carbon monoxide in expired air, 343 
Carbonic acid, 341 
Carnine, 326, 334, 344 
Caseic acid, 23, 46, 52 
Charcot-Neumann, crystals of, 330 
Cheese, poisonous, 52 
Chemotaxis, 129 
Cholera, 104 
Bujwid’s reaction, 110 
-blue, 112 
-infantum, 133 
-red, 110 
-stools, 213 
Choline, 34, 107, 237 
decompositions of, 243 
-group,232 
constitution of, 252 
Chorea gravidarum, 361 
Cicuta virosa, 176 
Codeine-like substances, 179 
Cod-liver oil, bases from, 263 
Comma bacillus, ferments produced 
by,104 
products of, 107 et seq. 
Colchicine-like substances, 181 
Zeisel’s test for, 182 
Collidine, 28, 196, 198 
Coniine, difficulties in detection of, 
177 
-like substances, 30,174 
Coridine, 204 
Corindine, 202 
Corn-meal, ptomaines in, 33, 83, 178 
Creatine, 189, 226 
Creatinine, 189, 226 
-group, 333 
Cruso-creatinine, 336 
Cyanogen, role of, 336 
Cystinuria, bases in, 207 
DE CONHSTCK’S bases, 198, 202 
Delezinier’s base, 204 
Delphinine-like substances, 180 
Deutero-albumose, 103 
-myosinose, 156 
Dialysis, concentration by, 172 
Diamines, 204 
Diarrhosas of infancy, 133 
Diethylamine, 191 
Digitaline-like substances, 28,179 
Dihydrolutidine, 195 
Dimethylamine, 34, 188 
Dimethyl-xanthine, 336 
Diphtheria, 124 
bacillus of, products of, 124 
immunity to, 128 
Dippel’s oil, 198 
Diseases, classification of. 84 
relation of bacterial poisons to, 84 
DragendorfFs method, 161, 167 
Dyspepsia, 356 
EBERTH’S bacillus, 16, 139 
products of, 140 
Eclampsia, 362 
Eel, poisonous, 41 
Ehrlich’s reagent, 260 
Enzymes, 35, 105, 118 
Ethylamine, 191 
Ethyleneimine, 205, 330 
Ethylidenediamine, 34, 204 
Expired air, leucomaines in, 341 
FECES, poisons in, 360 
Ferments, 35, 105, 118 
from comma bacillus, 104 
in urine, 357 
Fever, aseptic, 361 
of exhaustion, 359 
of fatigue, 358 
of non-elimination, 360 
Fish, poisonous, 41, 350 
Foods containing bacterial poisons, 36 
GADININE, 258 
Gaduine, 264 
Gaduinic acid, 263 
Galactine, 348 
Gautier’s pseudo-xanthine, 328 
Gautier and Etard’s bases, 199, 201, 
229 
methods, 163, 164 
extraction of leucomaines, 334 
German swine-plague, 142 
Germs, relation of, to disease, 85 et seq. 
Gerontine, 329 
Globulins, germicidal properties of, 
155 
Glucosines, 223 
Glycol, 190 
Goose-grease, poisonous, 51 
Gram’s bases, 273 
Griffith’s base, 203 389 
INDEX. 
Guanidine, 227, 312 
Guanine, 308 
Guareschi’s base, 268 
and Mosso's bases, 201, 273 
HAM, poisonous, 47 
Hankin’s method, 171 
Heteroxanthine, 319 
Hexylamine, 194 
Historical sketch of the bacterial poi- 
sons, 22 
Hog-cholera, 142 
-erysipelasj 142 
Homo-piperidinic acid, 231 
Hydrocollidine, 200 
Hydrocoridine, 204 
Hydrocyanic acid, 283, 354 
Hydrolutidine, 195 
Hyoscyamine-like substances, 27 
Ilypoxanthine, 298 
ICE-CREAM, poisonous, 79 
Immunity from blood serum, 
146, 147 
methods of securing, 146 
-producing substances, nature of 
of, 146 et seq. 
by intoxication with ptomaines, 
225 
to diphtheria, 128 
to pneumonia, 145 
to swine-plague, 144 
to tetanus, 119 
Indol, 111 
Infectious diseases, 84, 101 
how produced, 85 
definition of, 92 
favored by bacterial pro- 
ducts, 151 
Iso-amylamine, 193 
Iso-cyanacetic acid, 351 
Iso-propylamine, 193 
KAKKE, 41 
Koch’s rules, 92 
IACTIC acid, 106 
J Lactochrome, 348 
Lecithin, decomposition of, 240 
preparation of, 239 
Leucin, 19, 103, 109 
Leucocythfemia, urine in, 284 
Leucomaines, bibliography of, 381 
chemistry of, 280 
extraction of, 334 
pathological importance of, 354 
tables of, 351 
Lutidine, 195 
Lysatine, 189, 242, 333 
Lysatinine, 189, 242, 333 
Malignant oedema, 145 
Marino-Zuco’s method, 159 
Meal and bread, poisonous, 83 
Meat, poisonous, 50 
Methylamine, 187 
carhylamine, 351 
guanidine, 34, 108, 144, 225, 362 
hydantoin, 226, 340 
method of extraction, 167 
uramine, 226 
xanthine, 314, 319 
Milk, leucomaines in, 348 
poisonous, 62 
Monamines, 187 
Morin’s base, 222 
Morphine-like substances, 178 
Morrhuic acid, 263 
Morrhuine, 228 
Muscarine, 34, 251 
Mussel, poisonous, 36 
Mutton, poisonous, 51 
Mycoderma aceti, 16 
Mycoprotein, 19 
Mydatoxine, 34, 253 
isomer of, 255, 267 
Mydaleine, 34, 270 
Mydine, 34, 230 
Mylitotoxine, 34, 40, 255 
'VT ARCOTIC substance of Panum, 25 
i_l Nencki’s base, 196 
Neuridine, 34, 218 
Neurine, 34, 232 
Nicotine-like substances, 177 
Nicotinic acid, 199 
Non-toxicogenic bacteria, 13 
Nucleins, 141 
OSER’S base, 229 
Oxy-betaines, 265 
Oxygenated bases, 230 
Oysters, poisonous, 41 390 
INDEX. 
PANUM’S narcotic substance, 25 
putrid poison, 24 
Paraffin oil, bases in, 223 
Parareducine, 344 
Parasitic bacteria, 13 
Paraxanthine, 321 
Parvoline, 201 
Pellagroceine, 178 
Pentamethylenediamine, 213 
Pepsin, action of, 357 
Peptones, poisonous nature of, 354 
et seq. 
Peptotoxine, 275, 356 
Petroleum, bases m, 223 
Peptotoxine, 275, 356 
Phenyl-ethylamine, 197 
Phlogosine, 274, 129 
Phosphorus-containing substances,31 
Phytalbumose, 350 
Piperazine, 332 
Piperidine, synthesis of, 213 
Pneumonia, chemical products in, 
145 
Poisonous foods, 36 
Pouchet’s bases, 265, 268, 344 
Propylamine, 193 
Protalbumose. 103 
Protamine, 332 
Protomyosinose, 156 
Pseudo-xanthine, 328 
Ptomaines, bibliography of, 364 
chemistry of, 187 
definition of, 15 
table of, 278, 279 
separation of alkaloids from, 186 
methods of extraction of, 157 
remarks upon, 165 
Ptomatropine, 179 
Puerperal fever, 145 
Putrefactive alkaloids, 15 
Putrescine, 34, 107, 206 
Putrid poison of Panum, 24 
Pyocyanine, 277 
Pyogenetic proteids, 130 
Pyoxanthose, 277 
Pyridine, 107, 199, 202, 203, 275, 344 
RABBIT septicaemia, 144 
Reagents, purity of, 158 
Reducine, 344 
Reus’s test tor atropine, 179 
Rouget, 142 
Roussin’s test for nicotine, 177 
O ALAMANDARINE, 350 
O Saliva, leucoma'ines in, 346 
Salkowski’s base, 231 
Saprophytic bacteria, 13 
Saprine, 34, 220 
Sarcina botulina, 46 
Sarcine, 298 
Sarcosine, 340 
Sausage, poisonous, 22, 42 
Schweineseuche, 142 
Sebacic acid, 22,46, 52 
Selmi’s method, 27, 159 
Sepsine, 26 
method of extraction, 169 
Septicajmia of rabbits, 144 
Septicine, 194 
Sinapin, 242 
Spasmotoxine, 117, 194 
Spermine, 205, 330 
Spleen, leucoma'ines in, 347 
Staphisagria, 180 
Staphylococcus pvog. aureus, bases 
from, 274 
Stas-Otto method, 158, 167 
Stercoraemia, 360 
Strychnine-like substances, 32, 178, 
204 
reactions, 33, 1 fjO 
Sucholotoxine, 144 
Summer diarrhoeas of infancy, 133 
Suppuration, 129 
Susotoxine, 143, 223 
Swine-plague, American, 142 
products of bacillus of, 143 
German, 142 
TETANINE, 34, 117, 265 
Tetanizing substance, 32 
Tetanotoxine, 117, 194 
Tetanus, 113, 147 
bacillus, products of, 117 et seq. 
immunity to, 119 
neonatorum, 115 
toxines 194, 195, 255, 265, 267 
Tetrahydronaphthylamine, 201 
Tetramethylenediamine, 209 
Tetramethyl-putrescine, 210 
Theine, 316 
Theobromine, synthesis of, 316 
Theophylline, 326 
Toxalbumins, 19, 35, 118, 121, 127 
Toxicogenic bacteria, 13, 99 
Toxicology of ptomaines, 174 
Toxines, 15 
Toxopeptones, 109 INDEX. 
391 
Triethylamine, 192 
Trimethylamine, 34, 189 
Trimethylenediamine, 108, 205 
Tuberculin, 120 
Tuberculosis, 120 
products of bacillus of, 121 
Typhoid bacillus, 139 
products of, 140 
fever, 139 
Typhotoxine, 34, 140, 259 
isomer of, 258, 261 
Tyrosin, 103, 109, 197, 281 
Tyrutoxicon, 34, 41, 56, 61, 79, 269 
in summer diarrhcea, 139 
UNDETERMINED leucomaines, 
341 
ptomaines, 269 et seq 
Uraemic poisoning, 361 
Urea, 227 
Uric acid, 318 
group of leucomaines, 282 
Urine, ferments in, 357 
leucomaines in, 343 
toxicity of, 345 
Urochrome, 344 
Urotheobromine, 343 
VALERIANIC acid, 231 
Vanilla, 79 
Veal, poisonous, 51 
Venoms of serpents, 348 
Veratrine-like substances, 180, 204 
Vernine, 309 
Vitiated air, effects of inhalation of, 
362 
TTT'EIDEL’S reaction, 288 
VV White liquefying bacterium, 
products of, 139 
XANTHINE, 313 
group, constitution of, 318 
Xantho-creatinine, 337 
test for colchicine, 182