






    THE
    TOXINS AND VENOMS
    AND THEIR ANTIBODIES

    BY
    EM. POZZI-ESCOT

    AUTHORIZED TRANSLATION
    BY
    ALFRED I. COHN, PHAR. D.

    _FIRST EDITION_
    FIRST THOUSAND

    NEW YORK
    JOHN WILEY & SONS
    LONDON: CHAPMAN & HALL, LIMITED
    1906




    Copyright, 1906
    BY
    ALFRED I. COHN

    ROBERT DRUMMOND, PRINTER, NEW YORK




INTRODUCTION.


Our knowledge of the toxins is of quite recent date. It is hardly
twenty years since we began to acquire a knowledge of the facts that
are detailed in this volume, and to which modern medicine owes its most
recent and marvelous progress, particularly in serotherapy.

In this volume we have studied, besides the true toxins--substances of
cellular origin and of albuminoid nature and unknown composition--other
toxic substances, the nitrogenized alkaloidal bases introduced into
science through the researches of Selmi, Armand Gautier, and von
Behring, and which are highly hydrogenized nitrogenous crystallizable
principles of definite chemical composition--the products of the more
or less advanced breaking down of albuminoids.

Although these principles differ widely, by reason of their
physiological properties as a whole, from the toxic albuminoids, or
true toxins, it appears proper to consider them as products of the
advanced decomposition of these toxins--and in this respect their study
becomes imperative, the more so as they are very frequently encountered
together with the toxins, particularly in serpent-venoms, where their
action is exerted in addition to that of the true toxins.

In the first volume of this collection we dwelt on the essentially
reducing nature of the cellular functionation. To this
functionation--causing the splitting up or decomposition by hydrolysis
of nitrogenized albuminoid foods--is due the formation of these toxic
basic products within the organism, whether normally, or because of
certain pathological conditions.

This alone suffices to show that, during physiological life, oxygen
plays an essentially antitoxic rôle within the organism.

It is hoped that this succinct résumé, which it has been sought to
make as clear as possible, will be of service to those who, while
not scientists actively engaged in scientific progress, desire to be
abreast of the knowledge of modern evolution, but yet are not in a
position to consult original papers or large treatises.




CONTENTS.


                    PAGE

 INTRODUCTION                                iii


 PART I.

 _GENERALITIES REGARDING TOXINS AND
 ANTITOXINS._

 CHAPTER I.

 ALKALOIDAL TOXINS, PTOMAINES, AND LEUCOMAINES.

 Alkaloidal products of cellular life          1
 Ptomaines                                     4
 Physiological action                          5
 Extraction                                    5
 Classification, etc.                          7
 Leucomaines                                  10
 Xanthic leucomaines                          12
 Creatinic leucomaines                        13
 Neurinic leucomaines                         13
 Indeterminate leucomaines                    14

 CHAPTER II.

 TOXINS AND ANTITOXINS.

 Toxins                                       15
 Action of pathogenic bacteria                16
 Action of toxins                             17
 Nature of toxins                             18
 Origin of toxins                             20
 Autointoxications                            21
 General mode of action                       23
 Constitution of toxins; Ehrlich's theory     24
 Means of defense possessed by the organism
 against the action of toxins                 28
 Pasteur's vaccination method                 30
 Virus action                                 30
 Phagocytosis                                 32
 Antitoxins                                   33
 Mode of action                               35
 Formation; Ehrlich's theory                  38
 Serotherapy                                  41


 PART II.

 _THE TOXINS PROPER._

 CHAPTER III.

 I. VEGETABLE AND ANIMAL TOXINS.              42
 Abrin                                        42
 Ricin                                        44
 Robin                                        45
 Toxicity of the vegetable diastases          45

 II. TOXINS FROM MUSHROOMS                    46
 Phalline                                     48
 Symptomatology                               49
 Antidiastases                                51

 III. ANIMAL TOXINS                           53
 Peptotoxin                                   53
 Alimentary Intoxications                     55
 Urinary toxins                               57
 Variation of urinary toxicity                59
 Autointoxications (animal)                   60
 Glandular secretions                         62
 Suprarenal capsules                          63

 CHAPTER IV.

 THE MICROBIAL TOXINS.

 Pyogenic and pyretogenic properties          66
 Anthrax toxin                                67
 Tubercular toxin                             69
 Diphtheria toxin                             71
 Tetanus toxin                                76
 Mallein                                      79
 Typhoid toxin                                80
 Cholera toxin                                82

 CHAPTER V.

 THE VENOMS.

 General nature of venoms                     85
 Venomous serpents                            87
 Nature of serpent-venoms                     88
 Natural immunity towards serpent-venoms      90
 Artificial immunity towards serpent venoms   91
 Venoms of batrachians and saurians           92
 Fish-poisons                                 95
 Poisons of the hymenoptera                   96
 Poisons of scorpions                         97
 Poisonous blood and serums                   98
 Poisonous meats                             100




TOXINS AND VENOMS.




PART I.

_GENERALITIES REGARDING TOXINS AND ANTITOXINS._




CHAPTER I.

ALKALOIDAL TOXINS, PTOMAINES AND LEUCOMAINES.


=Alkaloidal Products of Cellular Life.=

Before entering upon the study of the true toxins, which are products
of an alkaloidal nature and of unknown composition, it is necessary to
say a few words regarding the most definite of the toxic alkaloidal
principles that are frequently encountered under various conditions,
conjointly with the true toxins, particularly in venoms, and which,
furthermore, are closely allied to these albuminoid toxins.

These principles are formed in essentially reducing media, whether it
be within the body of the organism, and by the simple exercise of its
normal function, in which case the principles bear the generic name
_leucomaines_[1]; or whether due to the action of anaerobic microbes,
when they are designated as ptomaines.[2] These basic principles, which
are essentially the products of cellular secretion, are usually toxic,
and sometimes even extremely so.

 [1] ARMAND GAUTIER: Sur les leucomaines, nouveaux alcaloides, dérivés
 de la transformation des substances protéiques des tissus vivants.
 _Bull. Soc. Chim._, 2e série, XLIII, p. 158.

 [2] ARMAND GAUTIER: "Communication sur les bases d'origine
 putréfactive." _Bull. Soc. Chim._ (2), XXXVII, p. 305.

As we shall presently see, ptomaines are essentially products formed
during putrefactive fermentation. The toxic properties of extracts from
the cadaveric fluids have long been known. Already in 1838 Panum[3] had
met with these products in snake venoms. Bergmann and Schmiedberg[4]
in 1868 isolated from septic pus a toxic substance which they named
_sepsin_; and almost at the same time Zuelzer and Sonnenschein[5]
reported having isolated from anatomical preparations an alkaloid
possessing mydriatic properties. It is, however, due particularly to
the researches of Selmi and Armand Gautier that we are now so well
informed regarding these toxic principles.

 [3] _Virchow Archiv._, X, p. 301.

 [4] _Medic. Centralblatt_, 1868, p. 497.

 [5] _Berlin. Klin. Woch._, 1869, No. 2.

The labors of Armand Gautier were first published in his _Traité de
Chimie Appliquée à la Physiologie_; those of Selmi in the _Actes de
l'Académie de Bologne_.

At first sight, there appears to be a great difference between these
alkaloidal bases, the ptomaines and leucomaines, and the albuminoid
toxins proper. The toxic bases of the first two groups are quite
definite chemical products which can be generally obtained quite pure,
and frequently in crystalline form. The toxins proper, on the other
hand, are highly complex albuminoid substances which greatly resemble
the true diastases in all their properties.

Nevertheless, between the toxic alkaloids, ptomaines and leucomaines,
and the toxic albuminoids, or more properly toxins, there exists no
absolutely sharp line of demarcation, but there is a gradual passage
from the one to the other by every intermediary grade, as a result of
the breaking down of the albuminoid molecule.

We shall see, moreover, as we proceed, that these substances are
formed under coexistent circumstances, and that they are, hence, found
together, whether it be in virus or in snake venom.

We will first consider the ptomaines, and then the leucomaines.

=Ptomaines.=

This name is more specially reserved to designate those alkaloidal
substances, generally highly hydrogenized, that are formed outside
the organism, from the fermentative action of anaerobic microbes on
albuminoid substances.

These bases are generally volatile, with an intense and tenacious
purulent odor; often, however, they possess a floral odor (aubépine,
syringa), and even like that of musk. They combine readily with acids
and with the chlorides of the heavy metals, yielding crystallizable
salts.

The ptomaines afford no specific reaction whereby they may be readily
identified; and their identification is effected only after a
painstaking analysis.

We must here call attention, however, to several of their more common
properties, beginning with their basic character, their oxidizability
by the air and consequently their well-defined reducing power--a
property that led Selmi to propose a mixture of ferric chloride and
potassium ferricyanide as a reagent for their detection.[6] They are
precipitated by all the general reagents for the vegetable alkaloids.
Selmi has given several reactions, such as those afforded by sulphuric,
hydrochloric, and nitric acids, which appear, however, to apply much
more to the impurities present than to the bases themselves.

 [6] Sulle ptomaïne od alcaloïdi cadaverici. Bologne, CLXXXVII, p. 11.

The physiological action of these bases varies greatly; in some
the action is an extremely toxic one, as in the case of neurine
and muscarine, which are true ptomaines; there are others, such as
cadaverine and putrescine, which are quite innocuous. The physiological
action of these bases, like that of the true toxins, is studied by
making hypodermic injections of solutions of the bases in healthy
animals, such as guinea-pigs, rabbits, and dogs.

In animals, the principal phenomena observed by Selmi to follow the
injection of the substances are the following: At first dilatation
of the pupil, then constriction; tetanic convulsions, soon followed
by muscular relaxation, and retardation, rarely acceleration, of
heart-beat; absolute loss of cutaneous sensibility; loss of muscular
contractility; paralysis of the vasomotors; greatly retarded
respiration; stupor, followed by death with the heart in systole.

It must be observed that in a number of cases where toxic researches
had been made in the past, these bases had been mistaken for poisons
which were believed to have been introduced into the organism with
criminal intent. No one will ever know how many have fallen victims in
the past to ignorance regarding the cellular mechanism!

The extraction of these bases is a tedious and difficult operation.
The materials must first be exhausted with water slightly acidulated;
then, after precipitating the albuminoids by boiling and defecating by
adding lead acetate, the liquid is evaporated to one-half its volume
and dialyzed in a vacuum.[7]

 [7] ARMAND GAUTIER: _C. rend. de l'Académie des Sciences_, CXIV, p.
 1256. _Ibid._, XCVII, p. 264, and XCIV, p. 1600.

Phosphomolybdate is then added to the dialyzed liquid, and the
precipitate formed, which now contains all the bases, decomposed by
boiling with lead acetate. After removing the excess of lead, there is
thus obtained a limpid solution of all the alkaloidal bases in the form
of acetates. These are separated by alcohol and by means of fractional
precipitations with various metallic salts, depending upon the known
properties of the bases.

In order to facilitate their study, the ptomaines have been grouped
under two distinct classes, the one embracing the cadaveric or
putrefactive ptomaines, of undetermined microbial origin, the other
containing the ptomaines formed by microbes of known character. Each
of these two groups is itself divided into subgroups, as shown in the
following table:

    GROUP I.

=CADAVERIC PTOMAINES OF UNDETERMINED MICROBIAL ORIGIN.=

  _a._ Amines.
  _b._ Guanidines.
  _c._ Oxamines (fatty or aromatic).
  _d._ Amido Acids.
  _e._ Carbopyridic Acids and analogues.
  _f._ Undetermined Ptomaines.


    GROUP II.

=PTOMAINES OF KNOWN MICROBIAL ORIGIN.=

  _a._ Ptomaines extracted from microbial cells.
  _b._ Ptomaines from pathological urines.

We will not here enter upon a detailed study of the bases belonging
to each of these groups. This subject is a vast one, requiring for
its treatment a volume devoted to it alone. We will here simply touch
upon the principal properties of several of the bases of each of the
subgroups named.


    BASES OF GROUP I.

_a._ =Amines.=--Among these we find nearly all the fatty amines, such
as the methylamines and the cyclic alkaloids such as pyridine. They are
formed particularly by the putrefaction of fish.

Certain of these bases are very toxic, for instance trimethylene
diamine, the collidines, and the parvolines.

_b._ =Guanidines.=--Among the products of ordinary putrefaction there
has been found so far only methylguanidine, C{2}H{7}N{3}. This is a
highly toxic base of which 0.2 Gm. is fatal to a guinea-pig.

_c._ =Oxamines.=--Under this designation the following bases are
comprised: 1. Neurine bases; 2. oxygenized aromatic bases; 3. bases of
unknown constitution. Amongst them we find neurine and choline, which
are toxic, and betaine, which is innocuous. They are found particularly
in putrid fish.

_d._ =Amido Acids.=--These ptomaines, which are usually innocuous in
small quantities, are particularly the products of the decomposition
of albuminoid substances. Among them we find glycocoll, leucine, and
tyrosine, as members of this group.

_e._ =Carbopyridic and Carboquinoleic Acids.=--So far only one base
is known belonging to this group, and that is morrhuic acid, which is
found in the decomposed livers of codfish, and which is a powerful
appetizer and stimulant in disassimilation.

_f._ =Undetermined Ptomaines.=--Under this heading are classed certain
undetermined bases, such as those found in normal urines, and in
spoiled meats and bread.

    BASES OF GROUP II.

_a._ =Ptomaines Isolated from Cultures of Pathogenic
Bacteria.=--Bacterial cultures contain, besides the true toxins, a
certain number of alkaloidal bases which sometimes possess considerable
toxicity.

In the cultures of streptococcus pyogenes there are found
trimethylamine and xanthic bases; in those of staphylococcus pyogenes
aureus are found xanthic bases and creatinine; while pyocyanine and
pyoxanthine are found in the cultures of bacillus pyocyaneus, etc.

_b._ =Ptomaines Isolated from Pathological Urines.=--Toxic ptomaine
bases have been found in the urines of a large number of diseases.[8]
It is quite probable that these bases are the results of a general
pathological condition due to some bacterial disease, the toxic
products of which are eliminated by the kidneys.

 [8] GRIFFITHS: _C. rend. de l'Académie des Sciences_, CXV, pp. 285 and
 667.

From the urines of epileptics Griffiths[9] isolated a colorless base
crystallizing in prisms having the formula C{12}H{15}N{5}O{7}, and
which was found to be exceedingly toxic; the same investigator isolated
from the urines of eczematous subjects a ptomaine which he named
_eczemine_,[10] and which is also highly toxic.

 [9] E. POUCHET: Contribution à l'étude des matières extractives de
 l'urine, _Thèse_, Paris, 1880; _Ibid._, _C. rend. de l'Académie des
 Sc._, XCVII, p. 1560; BOUCHARD: _C. rend. Soc. de Biolog._, Aug. 12,
 1882.

 [10] GRIFFITHS: _C. rend. de l'Académie des Sciences_, CXVI, p. 1206.

In certain cases of cystinuria there are found in the urine sulphurized
ptomaines, and in measles the urine contains an undetermined ptomaine,
_rubedine_, which is very poisonous. _Typhotoxine_, a very toxic
ptomaine, has been isolated from the urine of typhoid patients;
_erysipeline_, a hardly less toxic base, exists in the urine of
erysipelatic subjects; while _spasmotoxine_, _tetanotoxine_, and
_tetanine_, exceedingly active alkaloids, are found in the urines of
tetanus patients.[11]

 [11] BRIEGER: Untersuchungen über die Ptomaine, dritten Teil, p. 93;
 _Berichte d. D. Chem. Gesellschaft_, 1886, p. 3159; 1887, p. 69.

As a general rule, all abnormal urines contain toxic bases; the kidneys
appear, in fact, to serve as a means of eliminating the toxic products
that form in large quantity whenever, and for whatever cause, the
organism ceases to functionate normally, whether it be as a whole, or
in any one of its parts.[12]

 [12] CHARRIN: _Les poisons de l'urine_: Encyclopédie Léauté.


    =Leucomaines.=[13]

 [13] ARMAND GAUTIER: _Bull. Acad. de Médecin_ (2), XV, p. 115.

The leucomaines are basic substances, nearly allied to the ptomaines,
but still more closely related to the ureides. They are formed
directly or indirectly by the breaking down of protoplasmal
albuminoids. The agents that effect the breaking down are the
hydrolyzing ferments of the economy. It is well to recall here that
these phenomena of hydrolyzation occur within the cell itself and in
a practically reducing medium, as we have already stated. The inmost
mechanism of these phenomena cannot here be detailed; it will be found
described by Armand Gautier in the _Chimie Biologique_, and in his work
_Chimie de la Cellule Vivante_.[14]

 [14] ARMAND GAUTIER: Leçons de chimie biologique. Published by Masson;
 _Ibid._, Chimie de la cellule vivante. Also published by Masson.

The extraction of these bases is an extremely delicate operation. It is
necessary to operate with a large quantity of substance, say 50 kilos.
The substance is finely chopped, then exhausted with twice its weight
of water acidulated with acetic acid (0.2 Cc. per liter) and containing
a trace of oil of mustard, which is intended to act as an antiseptic.
The albuminoids are precipitated by boiling, the solution then
filtered, evaporated in a vacuum at 60° C., and the bases extracted
with 95-per cent. alcohol.

The alkaloidal bases obtained in this manner are separated by
crystallization from alcohol or by various other chemical methods, the
description of which we will not enter upon here.

In order to facilitate the study of the leucomaines they are classed
under three groups, according to their chemical affinities. These
groups are as follows:

1. =Xanthic Leucomaines.=--The bases of this group appear to have a
composition resembling that of uric acid. When hydrolyzed, they yield
urea and guanidine. They are weak bases, and exhibit both basic and
weakly acid properties. They all possess the common characteristic of
being precipitated by copper acetate in acid solution with heat, and by
ammoniacal silver nitrate in the cold.

According to Kossel, these bases are derived from the nucleo-albumins
which are found in the cell nuclei, and which are, as we know,
substances rich in nitrogen and phosphorus.

Among the bases of this group may be mentioned _adenine_,
C{5}H{5}N{5}, which is obtained from infusions of tea.[15] This base
is non-toxic; it was discovered by Kossel,[16] and it crystallizes
easily.

 [15] KRUGER: _Bull. Soc. Chim._ (3), VIII, p. 687.

 [16] KOSSEL: _Zeitschrift für physiol. Chim._, X, p. 248; and _Bull.
 Soc. Chim._ (3), III, p. 239.

Some others of this group are:

_Guanine_, C{5}H{5}N{5}O, non-toxic, discovered by Unger;
_pseudo-xanthine_, obtained from muscular tissues; _sarcine_,
C{5}H{4}N{4}O, also but slightly toxic, discovered by Scherer;
_xanthine_, C{5}H{4}N{4}O{2}, which is found in many urines, and
which acts as a stimulant on the cardiac muscles; _paraxanthine_,
C{7}H{8}N{4}O{2}, a toxic base found in certain pathological
urines; _caffeine_ and _theobromine_, powerful diuretic bases; and
_carnine_, C{7}H{8}N{4}O{3}, from meat, a muscular stimulant like
caffeine.

2. =Creatinic Leucomaines.=--These have for their type guanidine; they
differ from the xanthic bases in that they are not precipitated by
copper acetate, but frequently are by ammoniacal silver nitrate. They
yield double salts with the chlorides of zinc and cadmium. To this
group belong _glycocyanine_, C{3}H{7}N{3}O{2}, and _glycocyanidine_,
C{3}H{7}N{3}O, both very toxic; _creatine_, C{4}H{9}N{3}O{2}, only
slightly toxic; _creatinine_, C{4}H{7}N{3}O; _lysatine_, which very
easily decomposes to form urea; _lysatinine_, _xanthocreatine_;
_arginine_, a vegetable base, etc.

3. =Neurinic Leucomaines.=--These have none of the characteristics of
the preceding bases; their type is neurine, a highly toxic base found
in the brain, nerves, and certain fish ova. These bases are sometimes
normally produced by the animal economy, and are also frequently the
result of microbic action. They are the result of the simple phenomena
of fermentative hydrolyzation of protagons and lecithins. Among these
bases are _choline_, a weak alkaloid, and _betaine_, which appears to
be non-toxic.

The former has the formula C{5}H{15}NO{2}; it was discovered
by Stocker. Wurtz synthesized it by combining trimethylamine and
glycol-monochlorhydrine, and treating the resulting hydrochloride with
silver oxide. Betaine, C{5}H{11}NO{2}, is found in beets; it was
discovered by Scheibler. Neurine is, chemically, trimethylvinylammonium
hydrate.

4. =Undetermined Leucomaines.=--Among these bases several are
important in more than one respect. For instance _spermine_, which is
found in the sperm, is a strong base possessing a powerfully dynamic
and tonic action on the nerves. It acts as an oxidizer. Spermine
was first obtained by Schreiner[17] from the sperm of mammifers in
which it occurs as a phosphate. It has the formula C{5}H{14}N{2}.
It was physiologically studied by Poehl, Tarchanoff, Weljaminoff,
and Joffroy.[18] _Plasmaine_, a toxic base found in the blood and
discovered by R. Wurtz,[19] has the formula C{5}H{15}N{5};
_protamine_, from fish milt, was discovered by Micocher.[20]

 [17] _Liebig's Ann. der Chemie_, CXCIV, p. 68.

 [18] _Journ. Soc. Phys. Chim. Russe_, 1893, No. 2; and _Bull. Soc.
 Chim._ (3), XII, p. 243.

 [19] _Leucomaines du Sang Normal_, Thèse, Paris, 1889.

 [20] _Joh reab. de Thiérchen_, 1874, p. 341; Picard, _Ibid._, p. 355.




CHAPTER II.

TOXINS AND ANTITOXINS.


We have already seen, in the preceding chapter, that the microbes
and the cells of various organisms are capable of secreting definite
products of a toxic nature to which the names "ptomaines" and
"leucomaines" have been given. Researches, which were begun scarcely
twenty years ago, have shown that, besides these crystallizable and
definite products, we meet with basic non-crystallizable substances of
unknown composition, possessing special toxic properties, sometimes
even of extreme violence. These substances have been named "toxins."

At first this generic name was extended toward indefinite basic organic
products that could be isolated from tissues and tumors both normal and
abnormal; later on, however, the name was applied to toxic substances,
equally indefinite, isolated from the culture media of microbes and the
active constituent of various venoms.

It is only since 1885, when Charrin called attention to them, that
investigations began to be made regarding them. In 1888 Roux and
Yersin,[21] in their beautiful researches on diphtheria, pointed out
the diastatic nature of the properties of the active albuminoid matter
existing in the cultures of the specific bacilli of this disease. From
that period, these products began to take a more and more prominent
place, from year to year, in the study of pathological affections, and,
by developing the knowledge of immunity, they have opened a new path to
the investigations of therapeutic technic.

 [21] ROUX and YERSIN; Mémoire sur Diphtérie. _Ann. Inst. Pasteur_,
 1888-1889.

It is due to the knowledge of these principles that we have learned
that the infectious microbes, far from acting as they were believed to
do only a few years ago, and which Pasteur strongly maintained to be
by vital parasitism--such as would be the case with the carbonizing
bacteria which, according to Pasteur, act by diverting the oxygen,
or causing capillary embolisms--owe their pathogenic action to the
toxic substances which are the products of their secretion, and which
spread throughout the organism, even though the microbe frequently is
localized in a very circumscribed spot, as in tetanus and in diphtheria.

The idea of intoxication by these products has now replaced the idea of
the direct action of the microbe on the elements or the liquids of the
organism.

The occurrence that takes place in diphtheria and tetanus is one of the
best examples to cite in support of this view.

Here, in fact, the pathogenic microbe is found only in a very limited
area in the organism attacked--the false membrane, in the case of
diphtheria, or frequently only a slight wound in the case of tetanus,
and the microbe becomes localized there only. Now, in both cases,
there are general phenomena of toxic effects. There must hence be a
diffusion of toxic substances which, distributed by the blood, affect
the different systems and exert a toxic action on the entire organism.

It must be observed that the toxins act as toxic agents only when in
a condition to be introduced into the circulation subcutaneously.
The cause of this innocuousness of the toxins when given per os
has frequently been studied. It appears to be quite probable that
the cause of the attenuation of the morbid properties is due to
the intervention of the digestive microbes. Such is the opinion of
Levaditi and Charrin[22]; it is also the conclusion that is to be
drawn from the experiments of Mme. Metchnikoff and of Calmette[23] on
the modifications undergone by a vegetable toxalbumin, abrin, and by
serpent venoms, when these toxalbumins are inoculated with the bacillus
subtilis chromogenus. Moreover, Charrin and Lefèvre,[24] on the one
hand, and Nencki, Sieber and Somanowsky,[25] and Carrière,[26] on the
other hand, have discovered that the digestive ferments, particularly
trypsin, destroy, even though but little, the toxins secreted by
the Loeffler and Nicolaier bacilli. This is practically contrary to
the opinion of Behring and of Rauson,[27] according to which the
innocuousness of the microbial poisons when administered per os is due
exclusively to the lack of absorption.

 [22] CHARRIN and LEVADITI: Le sort de toxines introduites dans le tube
 digestif. _Journal de Physiologie et de Pathologie Générales_, 1898, p.
 226.

 [23] Citing Metchnikoff.

 [24] _C. rend. de la Soc. de Biologie_, 1898.

 [25] _Centralblatt für Bakt._, 1898.

 [26] _C. rend. de la Soc. de Biologie_, 1899.

 [27] _Deutsche Med. Wochenschr._, 1898, No. 8.

=Nature of the Toxins.=--The molecules of the toxins are very nearly
like those of the diastases. Like these, the toxins appear to have
a very complex, and very unstable, internal structure. Their mode
of action frequently depends, as in the case of the diastases, upon
the medium in which they occur. Again, like the diastases, they are
generally destroyed by the action of sufficiently prolonged heat, but
less easily, for there are certain toxins that resist a temperature
of 100° C. for an indefinite period. They are, like the diastatic
albuminoids, insoluble in strong alcohol, and are precipitated from
their solutions on the addition of this reagent. They easily adhere
to precipitates that form in liquids in which they occur in solution,
and possess the remarkable property of diastases in that imponderable
masses produce considerable results.[28]

 [28] See POZZI-ESCOT: Les diastases et leurs applications, published by
 Masson, 1900; and _Traité de Physico-chimie_.

Although closely allied to certain alkaloidal bases, the toxins are
sharply distinguished by the remarkable fact that their action is never
immediate, but is always preceded by a period of incubation, which may
be quite long.

Like the alkaloidal bases, they appear to result from the hydrolyzing
breaking down of albuminoids and nucleo-albumins, and they appear to
be intermediary, from a chemical point of view, between these bodies,
the general characters of which they retain, and the alkaloids proper,
or ptomaines, to which we have called attention, and the principal
chemical and physiological properties of which they possess.

No absolutely precise knowledge is had regarding the chemical nature
and constitution of these remarkable substances. A number of analyses
of these substances have been published which, in general, permit no
definite conclusion to be drawn.[29] I have, however, elaborated
several speculative ideas regarding this subject.[30]

 [29] Regarding this see the works by KOCH and BRIEGER, _Deutsche
 Medicin. Wochenschr._, Oct. 22, 1891.

 [30] POZZI-ESCOT: Nature des Diastases. Published by J. Rousset,
 Paris, 1903. See also Recherches de la Nature Chimique des Diastases
 Oxydantes. _Revue génér. de chimie_, VII, pp. 129-136; and Aperçus
 sur la nature chimique des Diastases, _Bulletin de l'Association
 de Chimistes_, 1904, p. 769.--Propriétés Catalytiques de Quelques
 Diastases; _Ibid._, 1904, p. 1247.

We must here call particular attention to the ideas of Ehrlich
regarding the constitution of the toxins. According to this scientist,
their molecules contain two functional groups; the one, to which he
has given the name "haptophore," is that which enables the toxin to
attach itself to any cellular element whatever, and which it then
renders non-toxic by means of the other, or "toxophore," group. We will
particularize farther on regarding this very important conception.

=Origin of the Toxins.=--These toxic bodies result either as the
products of the secretion of microbial life, or as the result of the
normal functionation of cellular life in the higher vegetable or animal
organisms.

They are the direct products of life, and do not result, as was
formerly believed, from a more or less profound modification of the
more or less complex albuminoids that serve as a food for the various
species of microbes, or for the cellular elements.

The vegetable toxins are less numerous than the animal toxins.
They are met with, nevertheless, in almost all mushrooms which are
reputed or known to be toxic; the seed of the castor plant contains a
very toxic vegetable albuminoid, as is likewise the case with Abrus
precatorius (jequirity-bean), and certain others.

The true physiological toxins occupy a very important place in the
realization of the conditions that govern health, sickness, and death.

We will see later on that they are met with in quite large number
in the bladder, whence they are voided in the urine. Their number
varies considerably, according to diverse influences (waking, slumber,
eating, fasting, fatigue, oxygen, brainwork, health, disease, etc.).
It is necessary here to observe that the renal system serves for the
purification of the entire organism, and that in the case of normal
life we will find in the renal system a large portion of the products
of the cellular secretion of the organism, and among the number there
are found, as we know, a certain number of alkaloidal bases. We will
take up later the subject of urinary toxicity.

=Autointoxications.=--The toxins are also encountered, and often in
some number, in the muscular tissues and in the blood, particularly
in those of batrachians, mureids, and saurians. In the organism these
toxins, developed by the activity of the various cells, may cause
autointoxication whenever, for one cause or another, their normal
elimination ceases. "Although there are an infinity of diseases,"
remarked Prof. Bouchard, "there are but a few ways of becoming ill." Of
these ways that of autointoxication is the most frequent. "What else
is it, then," says Prof. Charrin, "in the last analysis, but to die
from affections of the kidney, the liver, the heart, the lung, etc., if
it be not to succumb because of the lack of oxygen, the accumulation
of carbonic acid, the influence of the numerous urinary poisons, the
action of acids, of salts, of biliary pigments, or the effect of
noxious principles, which the hepatic cell must normally destroy or at
least attenuate."

These autointoxications, always due to poor elimination of toxic
principles, toxins formed in very great number in the organism, and
which the normal modes of evacuation or destruction do not eliminate,
are always found to be the cause of all diseases, even those that are
manifested by attacks of the cerebro-spinal axis, and that exhibit
variously mania, insanity, symptoms of hyperexcitability, etc.

These autointoxications are controlled by the nervous system, and
the latter alone is the cause of a larger number of maladies than
is generally believed; in fact, if the mechanism of nutrition be
reduced to its most simple elements, it will be seen to consist of the
penetration of the foods, of the plasmatic principles, to the cells;
of their transformation within the interior of the cells, and finally
the rejection of all the matter that could not be utilized. It is the
nervous system that commands or dominates this mechanism, that controls
the taking-up of assimilable elements and the elimination of toxic
principles, the fruit of assimilation or disassimilation, and in such a
manner, in fact, that this same nervous system can, at its will, cause
starvation, or intoxicate.

The marvelous cures obtained by magnetic methods are due to no other
causes than favorable changes in the nervous system.

=General Mode of Action.=--The toxins, of whatever kind, always behave
like diastases, in the sense that their definite action appears to be
absolutely independent of their mass, and that imponderable quantities
suffice to cause serious morbid affections and profound modifications
in nutrition.

Koch has shown that tuberculin is capable of affecting 60 trillion
times its weight of the living human being. According to Vaillard one
milligramme of tetanus toxin will kill a horse weighing 600 kilos.
These two examples show what an enormous power the toxins possess.

My views regarding the manner in which diastases act I have developed
at length in my work _Nature des Diastases_. The close analogy between
these substances and the toxins, an analogy upon which, moreover,
I have dwelt at some length, permits me to refer the reader who is
desirous of fuller details to the small work just mentioned.

The mode of action of diastases resembles singularly closely that of
the catalytic substances, and we will admit, for the moment, that they
act by intermediary combination, resulting in their rapid decomposition.

We owe to Ehrlich[31] a new conception relative to the nature and mode
of action of the diastases, and which to-day plays an important rôle in
all our conceptions regarding immunity.[32]

 [31] EHRLICH: _Klinisches Jahrbuch_, 1897, VI. _Proceedings of
 the Royal Society_, 1900, No. 482, p. 424. _Nothnagles' specielle
 Pathologie und Therapie_, 1901, VIII, Schlussbetrachtungen, p. 163.

 [32] To have a complete exposé regarding this question, it will be
 profitable to consult No. 4 of this collection on _Sérums Immunisants_.

According to this scientist, the complex molecule of albuminoid
substances is constituted by a fixed central nucleus, and by a number
of lateral chains or receptors, fixed to this nucleus, which possess
diverse accessory functions, and which serve, particularly, for the
nutrition of the cells. These receptors have a great affinity for the
various substances necessary for the support of the living elements,
and they seize upon the alimentary substances, in normal life, just as
a leaf of the _Dionæa_ seizes a fly which serves as its food.

In these special conditions the receptors may attach themselves to
the complex molecules of albuminoid substances, such as the different
toxins.

Ehrlich supposes, as we have already seen, that a toxin contains two
special groups--a _toxophore_ group, which poisons, and a _haptophore_
group, which combines with the receptor. According to this theory,
the toxophore group of a toxin can act on an organism _only_ when the
haptophore group of the toxin encounters a suitable attachment or
receptor.

The receptors attached to the living protoplasmic molecule attract the
toxin, just as a lightning-rod attracts the lightning.

It is hence clearly proved that the toxigenic poisons exert their
noxious action on the cellular elements of sensitive organisms, by
entering into combination with these.

Experience has shown that they attach themselves, in a most rigorously
elective manner, to the tissues, and rapidly disappear from the general
circulation. Numerous facts, clearly established, attest the reality of
this fixation or attachment.

It is thus that von Behring and Wernicke[33] sought to ascertain
the quantity of antitoxin (we will see farther on that this name is
given to those substances which neutralize the activity of toxins
under certain conditions) which, introduced a certain time after the
introduction of the poison, will save the life of the animal. They
have experimented with diphtheria toxin, which we will study later,
and they have demonstrated that, if the antitoxic serum be introduced
immediately after the toxin, a dose of antitoxin twice as large as that
of the toxin suffices to effect a cure.

 [33] VON BEHRING and WERNICKE: Zeitschrift für Hygiene, XII.

Eight hours after the administration of the toxin the dose must be
trebled, while after thirty-six hours it is necessary to have recourse
to a quantity of antitoxin eight times as great. These experiments
show that the curative action of the antitoxin is so much the less the
longer the period of time that has elapsed between the introduction
of the toxin and the antitoxin. This is because the toxin has become
so intimately attached to the tissues that the antitoxin introduced
has not the power to destroy the combination. These facts have been
confirmed by Donitz[34] and by the classic experiments of Decroly and
Rousse.[35]

 [34] DONITZ: Ueber die Grenzen der Wirksamkeit des Diphtheria
 Heilserums. _Deutsche Med. Woch._, No. 27, 1897.

 [35] DECROLY et ROUSSE: _Arch. Int. de Pharmacodyn._, III and VI;
 Masoin: _Arch. Intern. de Pharmacodyn._, II, 1903.

 This is not, however, the case with cold-blooded animals, which,
 generally, are not affected by injections of poisonous toxins. Thus
 Metchnikoff[36] and his pupils have been able to show that the toxins
 introduced into certain cold-blooded animals (Oryetes nasicorius) may
 remain for several months without alteration in their circulation.

 [36] METCHNIKOFF: _L'Immunité_, Paris, 1902; MORGENROTH: Zur Kenntniss
 des Tetanus des Frosches. _Deutsche Med. Woch._, No. 35, 1898.

If we consider the facts of the theory of Ehrlich's lateral chains,
which we have mentioned, we are led to well-defined conclusions
regarding the mode of action of the toxins. In fact, since these toxins
exhibit a pronounced chemical affinity for the tissues, and while, on
the other hand, they can attach themselves only because of the presence
of certain functional groups of the protoplasmic molecules, this union
can take place only in certain specific centers. This has been fully
confirmed by experiments _in vitro_.

It is known, since the researches of Ehrlich,[37] Wassermann and
Takaki,[38] Marie,[39] Metchnikoff,[40] and a host of other scientists,
that this fixation is due to a clearly elective property. It is for
this reason that the tetanus toxin fixes itself only upon the nervous
tissue, and that in this action all passes as if the nervous tissue had
been provided with functional groups possessing an elective affinity
for the tetanic poison.

 [37] EHRLICH: _Berl. Klin. Woch._, No. 12, 1898.

 [38] WASSERMANN and TAKAKI: _Berl. Klin. Woch._, _Med._, p. 5, 1898.

 [39] MARIE: Sur les Propriétés Antitoxiques aux Centres Nerveux de
 l'Animal Sain. _Ann. Inst. Past._, 1898, p. 1.

 [40] METCHNIKOFF: Recherches sur l'Influence de l'Organism sur les
 Toxines. _Ann. Inst. Past._, 1899, p. 82.

=Means of Defense Possessed by the Organism against the Action of
Toxins.=--We have already seen that the renal organs serve for the
elimination of the toxins normally produced in the organism by the
simple play of its cellular mechanism. Experience has shown that the
toxins introduced from without into the circulation are generally
finally eliminated, even though in the meantime the modifications they
have imprinted on the economy may be transmitted hereditarily; and that
their influence on the general nutrition and the normal functionation
of the entire organism persists even after their elimination.

Much has been said regarding the elimination of these toxins by the
urine, but the experiments made by Métin, at the Institut Pasteur, have
shown the inaccuracy of this assumption, and it has been necessary to
seek another.

It has been remarked that oxidation destroys the toxins _in vitro_, and
it has been thought that a process resembling disinfection may well
take place within the tissues of the animal economy, but no decision
has been arrived at regarding the possible mechanism of this action,
which some attribute to the action of the oxidizing ferments of the
organism, or to the action of certain special cells.

According to Poehl, there is developed as destroyer a substance
possessing energetic oxidizing properties, which he has isolated and
named _spermine_, and which is found in most of the organic fluids
and particularly in the leucocytes, the special rôle of which we will
presently study.

There develops still another cause of elimination, or, to be more
exact, of the neutralization of the toxic principles in defense of the
organism against the toxins, and that is the formation of _antitoxins_.

It is well known that the term _virus_ has been reserved to designate
physiological liquids which were characterized, when first they were
known, by their property of transmitting to an organism certain
functional affections, but the true character of which is to expend
their toxicity upon the microbes which occur and are reproduced in the
organism, or upon the organized plastidulary granulations, as in the
case of the rabic virus, the special microbe of which has not as yet
been isolated.

Pasteur, when studying rabies, found that the brain and spinal marrow
of rabid animals contained the pure rabic virus in considerable
quantity, and that every particle of the marrow was capable of
imparting rabies to a perfectly healthy dog. After having ascertained
this fact, he found that he could _attenuate the action of the virus_,
either by passing the virus through certain animal organisms, such
as the monkey or rabbit, by gently heating, or even by allowing it to
oxidize and partially dry in the air, or else by submitting it to the
action of antiseptics or alternating electric currents of very high
tension.

Experiments have shown that a deadly virus, attenuated by one of the
means mentioned, may be injected, without danger of death, into the
living animal; and what is still better, the animal thus treated
acquires the power of resisting large doses of the virus, less and less
attenuated, and that it is possible to reach a point where the animal
economy may become habituated to very large doses of a highly virulent
virus without the organism experiencing any visible illness--that is,
the organism has been _vaccinated_ with regard to the particular virus.

Experiments have shown that this property is not peculiar to microbial
virus alone, but that it is common to the venoms the toxicity of which
is essentially due to some toxins, with the exception of those agents
noted.

The attenuated viruses act, as vaccins, through their soluble
constituents, which, either directly, by modifying the nutrition of
certain cells, or indirectly, by inducing reactions of the nervous
centers which preside over this nutrition, profoundly change the
conditions of life and give rise to the pathological condition--the
vaccined state.

Experiments by Behring and Kitasato[41] have shown that the tumors of
a vaccinated animal, freed from all organized matter visible under the
microscope by filtration through porcelain, contains principles capable
of directly or indirectly protecting other animals from the disease
caused by the corresponding virus. Meanwhile, experiments have shown
that the vaccinating matters are totally eliminated; nevertheless,
after their elimination, the immunity acquired remains with the animal,
which then continues to be protected against the corresponding virus.

 [41] _Deutsche Med. Wochenschr._, 1890, p. 1113.

Interest in this subject has incited numerous researches with a regard
to bringing to light the mechanism of this immunization; and this will
form the subject of another volume of this collection. We may state
here, however, that there have been recognized two concurrent causes of
this preservative action; the one, called _phagocytosis_, results from
the fact that the microbe introduced into the vaccined organism becomes
incapable of producing its usual toxins, while on the other hand the
immunization renders the organism capable of secreting substances
possessing an activity contrary to that of the virus, in fact true
counter-poisons, comprised under the general name _antitoxins_.

=Phagocytosis.=--We have seen that an organism subjected to a toxic
invasion tends to protect itself by proper means of defense; and one
of those is the direct putting into activity of the living cellular
elements themselves, and in particular, the leucocytes, or white
corpuscles, found in more or less number, according to pathological
conditions, in the blood and lymphatic fluids.[42]

 [42] It is necessary here to consult the work by LEVADITI: Le Leucocyte
 et ses Granulations. _Scientia_, Naud, publisher, Paris, 1903; also
 METCHNIKOFF: L'Immunité, Paris, 1902, Masson, publisher.

Metchnikoff has shown that the moment a foreign element, particularly
a microbe, enters the organism, these leucocytes come flocking from
all parts of the body, collect around the bacterial element, penetrate
it, and begin to digest it. These elements have received the name
_phagocytes_. The name _chemotaxis_ has been given to the property by
virtue of which they approach (positive chemotaxis) or move away from
(negative chemotaxis) certain substances which affect them powerfully.

Experiments have shown that the leucocytes are attracted by the
products secreted by pathogenic microbes, or saprophytes. Attracted by
the latter, the white corpuscles surround, envelop, and finally digest
them; and when it happens that all the pathogenic microbes within an
organism are absorbed, the organism survives, while in the contrary
case it succumbs.

Attention must be called to this attack by the white corpuscles within
the limits where they are normally confined. It is a pathologic
diapedesis--a leucocytosis provoked by the irritation of the
tissues--and caused either by the presence alone of foreign elements,
or by the soluble products secreted by them.

When, for any reason whatever, this phagocytic action is impeded,
the resistance of the organism to pathogenic infection ceases to be
effective, and the organism may therefore be invaded by the microbe.
Numerous causes may contribute to impede this action.


    =The Antitoxins.=

We have seen that the second means of defense possessed by the organism
resides in the action of special products, true defensive secretions,
possessing an activity contrary to that of the toxins, and which are
secreted by the cells of the organism under the influence of the
vaccins.

This is a property common to every organism, and which is observed even
in non-vaccinated subjects, although in this case the secretion forms
with great difficulty and in small quantity.

When an organism subjected to the toxic action of a bacterial infection
does not succumb to the intoxication, it emerges from the test gifted
with a new property, which may be augmented by habituation, and which
borders on immunity.

At first we were content to vaccinate small animals in the laboratory,
but in proportion as the discoveries in this domain extended, and there
developed a need for large quantities of antitoxins, recourse was had
to the larger animals, particularly horses and cattle. From the moment
that large quantities of blood and antitoxic serum were at command,
search was made for a means of isolating the antitoxin and determining
its properties.

Experiments so far made have shown that the antitoxins are substances
of an albuminoid nature, of unknown composition, and which are very
closely united to the albuminoid substances of the serum. It must
be observed, however, that Behring and Knorr oppose the assertion
regarding the albuminoid nature of tetanic antitoxin, but their reasons
for this do not appear to be well founded.

In general, these antitoxins are precipitable with the globulins, and
possess quite considerable powers of resistance towards physical and
chemical agents. Thus they are destroyed only at a temperature above
60-65° C. Kept in the dry state, in the residue of evaporated serum,
and away from the light and all oxidizing action, it is possible to
preserve their activity for a very long time.

They are essentially humoral substances; they are found in the blood
of vaccinated animals, from which may be obtained antitoxic serums
with a specific but transient immunity; and they are also found in the
plasmas of the lymph and exudates, in aqueous tumors, and in the milk.
They are seldom found in the cells.

=Mode of Action.=--Frequent attention has been paid to the mode of
action of the antitoxins upon the toxins, a phenomenon of great
importance in relation to the phenomenon of immunity acquired against
the toxins. At the beginning of our knowledge on this subject, the idea
of a destruction of the toxin immediately suggested itself, and was
advanced by von Behring.[43] According to this scientist the antibody
inhibits the morbigenic action of the toxin by neutralizing the toxin,
combining with the latter to form a compound of a chemical nature which
is devoid of toxicity and without action on the organism. According to
this theory, the influence of the antitoxin on the toxin is direct, and
does not require the intervention of the living cellular protoplasm.
Such was also the belief of Prof. Ehrlich.[44]

 [43] VON BEHRING and KITASATO: _Deutsch. med. Wochenschr._, 1890, p.
 1113.

 [44] EHRLICH: _Klin. Jahrb._ 1897, VI, p. 292.

Buchner, a little later, believed that the antitoxin, instead of acting
directly on the toxin, exercised a direct influence on the living
elements of the organism, preserving them from intoxication.[45]

 [45] BUCHNER: _Münchener med. Wochenschr._, 1893, p. 480.

Such was also the opinion of Roux[46]; and Calmette demonstrated that a
mixture of venom and of a non-toxic antivenom recovered its toxicity on
being heated to 68° C, whereby the antivenom was destroyed (Calmette:
_Le Venin des Serpents_, Paris, 1897, p. 58); and Wassermann arrived at
the same result.[47]

 [46] ROUX: _Annales de l'Institut Pasteur_, 1894, VIII, p. 724.

 [47] WASSERMANN: _Zeitschr. für Hygiene_.

The array of proofs offered by these scientists, which we cannot here
enlarge upon without uselessly extending our subject, would tend to
make one believe, at first glance, that the antitoxin does not act
directly on the toxin, but at the present time Buchner's theory appears
untenable. Numerous researches have proved conclusively that the toxin
and the antitoxin have a specific affinity for each other, by virtue
of which these principles combine to form a substance free from all
toxicity, but unstable, and which may be decomposed by heat or certain
other factors.[48]

 [48] J. DANZSY: _Annales de l'Institut Pasteur_, XVI, p. 331.

Some recent experiments by J. Martin and Cherry (_Proceedings of the
Royal Society_, 1898, LXIII, p. 423) have clearly brought out this
fact. These authors made mixtures of serpent venom with its antivenom,
which they filtered through a layer of gelatin, under the supposition
that, if the venom and its antivenom were not chemically combined,
the former alone would be able to pass through into the filtrate,
because its molecules are so much smaller. Martin and Cherry allowed
the venom and its antivenom to remain in contact for varying periods
before filtering. As the result of a series of experiments carried
out with this idea, they have demonstrated that the filtrate obtained
after allowing a few minutes' contact between the two substances,
was decidedly toxic, while that obtained after a contact of half an
hour was absolutely non-toxic. From this the authors conclude that
the antitoxin enters into chemical union with the venom, but that the
combination does not take place immediately, and requires a certain
length of time for its accomplishment.

Ehrlich and Knorr have demonstrated that the neutralization is less
rapid in dilute solutions than in concentrated ones.

Prof. Svante Arrhenius has completed our knowledge regarding the mode
of combination between the toxins and the antitoxins, by demonstrating
the occurrence of limited reactions analogous to the etherification of
an alcohol by an acid, and in such a manner that there always exists,
in a mixture of these two substances, a certain quantity of free toxin
and antitoxin. This is an important modification of the general ideas
held in this respect.[49]

 [49] SVANTE ARRHENIUS: La Physico-chimie des Toxines et des
 Antitoxines. _Conférences de la Société chimique de Paris_, May 20,
 1904. See also MADSEN AND ARRHENIUS: Testkrift red indivulsen of
 Stotens Serum Institut. Copenhagen, 1902.

It appears necessary to bring here more clearly in evidence the fact
that _the antitoxin inhibits the noxious action of the toxin, even
outside the living organism, by uniting with it to form a compound in
identically the same manner as when a strong base and a strong acid are
brought together_. As we have seen, all the conditions of environment
that favor or retard the formation of salts, in a like sense influence
the neutralization of the toxin by its antitoxin.

=Formation of Antitoxins.=--Ehrlich's theory of side chains, to which
reference has already been made, furnishes us with an explanation of
the formation of the antitoxins in tumors. Let us suppose that, in
the organism, a cell had come into contact only with certain toxic
molecules incapable of compromising its life, and that the only result
was the immobilization of the receptors which are united with the
haptophore groups of the opposing toxins. It is known that, by virtue
of a property inherent in all living organisms, during the phenomena
of reparation, there is generally an overproduction of the neoformed
parts. In the case we here speak of, as the receptors fill an important
function in the nutrition of the opposing cellular elements, once they
become united with the toxic haptophores, they become incapable of
filling their normal function of nutrition. Under these conditions the
cells develop so large a quantity of receptors that, filling the cells,
and not finding any more room, they spread into the blood and other
liquids of the organism.

Under these conditions, every new injection of toxin into the organism
is absorbed into the blood where it meets with the free receptors which
possess great avidity for the haptophore group of its molecule, and the
two groups immediately unite, before the haptophore group of the toxin
has been able to attack and intoxicate a cellular element.

We thus see that the receptors which, when in a free state in tumors,
play the rôle of antitoxics or antitoxins, become, within the cellular
elements themselves, the vehicle of intoxications. Figuratively
speaking, so long as these fixators are attached to the molecule of the
living protoplasm they attract the toxin.

According to this ingenious conception, the formation of antitoxins is
hence absolutely independent of the action of the toxophore elements on
the cellular elements, and it suffices that these possess receptors or
side chains capable of uniting with the haptophore groups of the toxin.
This explains why it has been possible to produce antitoxins from
toxins which have lost some of their toxic properties, but which have
preserved their property of uniting with antitoxic substances. Ehrlich
gives the name _toxoids_ to those modified toxins that have lost their
toxophore groups, while the haptophore group, the producer of the
immunizing substance, is still preserved intact.

According to Metchnikoff's theory, which is very similar, it seems
quite possible that the phagocytes, thanks to the facility with
which they absorb poisons, occupy an important place as producers of
antitoxins. It has not been possible so far to verify this theory in
our at present imperfect knowledge regarding this subject. The domain
of immunity has, however, made brilliant conquests during these last
few years, so that we should not despair of arriving at a definite
solution before long.

In the vaccinated animal the antitoxin is reproduced, and it is
possible to obtain several times, from the vaccinated animals,
successive portions of antitoxic serum.[50] The protective power
of these antitoxins is absolutely marvelous. An animal accustomed
gradually to the tetanic virus yields a serum containing an antitoxin a
thousand times more active than the virus.

 [50] CH. SALMONSEN et TH. MADSEN: Réproduction de la substance
 antitoxique. _Ann. Inst. Pasteur_, XII, p. 762. ROUX et VAILLARD:
 _Ibid._, 1893, p. 83.

According to Vaillard, a quintillionth of a cubic centimeter of this
antitetanic serum suffices to preserve one gramme of living mouse from
the effects of a dose of tetanic serum that would otherwise be surely
fatal.

In the animal, the antitoxins are eliminated mostly by the fluids of
the body, and particularly by the urine. Ehrlich has demonstrated that
they also pass into the milk, and this fact is confirmed by a large
number of observers. It explains the immunity acquired by nurslings,
and which is transmitted by the milk.

=Serotherapy.=--The search for antitoxins and their rôle in the
etiology of infectious diseases are fundamental points in actual
therapy. It has been demonstrated that the serums of certain vaccinated
animals enjoy very extended antitoxic therapeutic properties; for
instance, the serum of vaccinated rabbits is an antivenom towards
erysipelas; and the sterilized cultures of the pneumococcus or of the
Bacillus pyocyaneus prevents infection of carbuncle (anthrax).

The antivenomous serum of the ass immunized by injections of increasing
doses of the venom of the terrible naja is a perfect prophylactic and
curative, not only as regards the venom of this serpent, but also
against that of the crotalus, trigonocephalus, and viper.

We shall take up the study of serotherapeutics in another volume of
this collection.




PART II.

_THE TOXINS PROPER._




CHAPTER III.


I. VEGETABLE AND ANIMAL TOXINS.

The vegetable toxins possess the characteristic property of being
innocuous, and of being almost completely devoid of poisonousness, when
they are absorbed by the intestines; we can see, from this, how greatly
they differ from the poisons proper.[51]

 [51] It is understood that the active principles of mushrooms are not
 comprised under this definition, but they will be studied under the
 next heading.

The vegetable toxins known are quite numerous; nevertheless our
knowledge regarding them is very incomplete. Our review of them will be
chiefly descriptive.

Many of the leguminous plants are poisonous, either because of
emanations exhaled by them, or by reason of their alkaloids, or because
of some toxins contained in them. We shall commence with these.

=Abrin.=--This toxin, which was studied in particular by Warden
and Waddell,[52] then by Kobert[53] and de Hellin,[54] is found
in the fruit of the Leguminosæ, Abrus precatorius (wild licorice,
or jequirity). Its name was given it by Warden and Waddell, who
discovered both its toxic nature and the vegetable toxin; the toxin
is found only in the seeds. To extract it, the seeds are macerated
in water, and the solution filtered and precipitated with alcohol;
the precipitate which forms is collected and dissolved in distilled
water, from which it is again precipitated by adding powdered ammonium
sulphate. The precipitate is then collected and submitted to dialysis
in order to eliminate the ammonium sulphate. The abrin so obtained
forms an albuminoid substance[55] stable at 100° C., and possessing
rotatory power; it liquefies starch paste, and is extremely toxic. One
milligramme suffices to kill a rabbit within several hours. It must
be observed, however, that, as is the case with all the toxins, abrin
acts or kills only after a period of incubation which generally exceeds
twenty-four hours.

 [52] WARDEN and WADDELL: _Non-bacillar Nature of Abrus Poison_.
 Calcutta, 1884.

 [53] KOBERT: _Arbeit. aus dem Pharmak. Institut._ Dorpat, 1893.

 [54] HELLIN: _Inaug. Dissert._ Dorpat, 1891.

 [55] EHRLICH: Experiment. Untersuchungen über Immunität. _Deutsch. Med.
 Woch._, 1891.

It is possible to vaccinate an organism so as to withstand a lethal
dose of abrin, but it requires quite a long time; it is effected by
injecting into a suitable animal very small doses of the substance,
and increasing the quantity gradually. Rabbits which have been
rendered highly immune towards venoms are capable of resisting without
inconvenience doses of abrin which are ordinarily fatal; and the blood
serum afforded by them contains a specific antibody for the substance.

=Ricin.=--This vegetable toxalbumin has been studied particularly by
Stillmark,[56] by Dixon,[57] and Thuson.[58] It is found in the seeds
of the castor plant; three or four of the seeds suffice to cause a
gastroenteritis accompanied by serious symptoms and even by death.

 [56] STILLMARK: _Arbeit. aus dem pharmacol. Inst. Dorpat_, 1889.

 [57] DIXON: _Austr. Med. Gazette_, 1887.

 [58] THUSON: _Journ. f. prakt. Chem._, XCIV, p. 444.

It was first isolated by P. Ehrlich, by treating the seeds with
lukewarm water, and precipitating the aqueous solution with alcohol.
The toxalbumin is soluble in water, but on boiling the solution, the
substance loses in great measure its activity.

Ricin possesses considerable activity. 0.00003 Gm. suffice to kill a
rabbit when injected hypodermically; 0.2 Gm. are fatal to man. The
action is not immediate, but follows a period of incubation. Ehrlich
has shown that, exercising precaution, it is possible to create, as
with abrin, a condition of tolerance or habituation, and in consequence
to cause the formation of a specific antibody.

=Robin.=--This toxic albuminoid was obtained from the bark of an Acacia
(Robinia Pseudacacia) by Power and Cambier,[59] by exhausting with
water at a temperature of about 30° C., and precipitating the infusion
with alcohol. The substance is analogous to ricin, and like this,
possesses powerful toxic properties.

 [59] POWER and CAMBIER: _Pharm. Journ. and Transact._, 1890.

=Toxicity of the Vegetable Diastases.=--The diastases, which have been
treated of in a volume of the Encyclopédie Léauté,[60] and to which
we would refer the reader who is desirous of more complete details,
develop powerfully energetic toxic properties when injected into
the organism. Thus _amylase_ causes, when injected subcutaneously,
a considerable rise of temperature, but without any other toxic
symptoms. _Invertin_ or _sucrase_ was studied by Roussy under the name
_pyretogenin_, but it appears probable that this diastase was not the
only substance present in the product, but that there were present
reducing diastases, as we have already shown in the first volume of
this collection, devoted to the phenomena of reduction within the
living organism.

 [60] POZZI-ESCOT: _Les Diastases et leurs Applications_, Masson, 1900.

The pyretogenin of Roussy gives rise to an attack of violent fever, but
it loses all activity when heated to 80-100° C.

Through his researches, Roussy clearly demonstrated,[61] for the
first time, that the fever may cause the formation within the blood
of a substance clearly belonging to the class of soluble ferments or
zymases. Now, it is well known that within the animal economy there
exist many ferments of this character; and experiment has shown that
they can, at a given period and under various influences, leave the
cells in which they are normally localized, pass into the blood plasma,
and reach the nervous centers, where they cause serious effects. We
have already dwelt upon the mechanism of autointoxication of the
organism. The toxic action of certain digestive diastases has been
shown by Hildebrandt, who has demonstrated that 0.1 Gm. of pepsin is
capable of killing a rabbit in two or three days.

 [61] ROUSSY: _Aperçu historique sur les ferments et fermentations_.
 Paris, 1901. J. Rousset, publ.


    II. TOXINS FROM MUSHROOMS.

Mushrooms are alimentary substances of the highest order, causing
a general stimulation of the entire organism. The substances
met with belong, according to their composition, to different
classes--celluloses, sugars, and amylaceous substances, alcohols,
acids, fats, astringents, essential oils, resins, alkaloids, and
albuminoids. The study of the last only, the albuminoids and diastases,
interests us here. The most important of these albuminoid substances,
_phallin_, was discovered in 1890 by Kobert. Pouchet also has isolated
a whole series of other toxic albuminoids, particularly from Amanita
muscaria (Fly Agaric).

There are alimentary as well as toxic species in every possible variety
among mushrooms, some species consisting chiefly of the edible kind,
others consisting of the poisonous variety.

In consequence of the toxicity of mushrooms, great attention must be
given to the treatment to which they are subjected when it is desired
to utilize them for alimentary purposes. Thus the toxic principles of
several varieties can be removed, and the mushrooms rendered edible by
very simple means.

Pouchet has made a very ingenious comparison between the ethereal,
alcoholic, saline, and aqueous extracts of mushrooms, and bacterial
cultures. The analogy is striking as to the presence of toxin,
toxalbumose, and albumoses more or less toxic; it is moreover not
exaggerated, since, according to the classification generally admitted,
mushrooms are nothing more than the very advanced representatives of a
group the more simple members of which constitute the bacteria.

The same author has shown that phallin obtained from the juice of the
Fly Agaric will kill a guinea-pig weighing 600 grammes in one hour.

As we have already stated, it is the phalline to which the ordinary
disorders which mushrooms cause are due. According to Kobert, a 1:250
000 solution of this substance causes an intense hemolysis, with all
its disastrous consequences.

According to Pouchet, the flesh of mushrooms must be compared with meat
that has been kept for some time to become tender, and it is well known
that though this "tendering" process renders the meat more digestible,
it may also allow the meat to acquire noxious properties, due to the
presence of toxins.

Phallin is the type of those toxic albuminoids of unknown composition
which exist in mushrooms, and which are comprised under the name
_sapotoxins_. The intravenous injection of phallin into an animal, in
the proportion of 1 part to 1 000 000 parts of body weight, causes
sudden death within one minute; in the proportion of 1:5 000 000, death
occurs in about three minutes; in the proportion of 1:50 000 000, death
also occurs, but is greatly retarded. An injection of 0.0005 Gm. per
kilo of body weight of animal causes solution of the blood corpuscles
to such an extent that thirty minutes later the blood serum is strongly
colored red, as well as the veins.

Instead of being easily altered under the influence of an elevated
temperature, as are many of the albuminoid substances, whereby their
toxic power is lost, phallin may be boiled for half an hour with water
without undergoing any noticeable alteration. Pellegrini has observed
that the dried juice of Amanita Phalloides (Death-cup) preserves its
properties for more than a year.

According to a recent paper by Gillot, the symptoms of poisoning by
mushrooms must be ascribed to albuminoids (phallin and albumose),
alkaloids (muscarine, choline, or betaine), or to resinoids (cambogic
and agaricic acids).

The _alkaloids_ found in mushrooms are: _Muscaridine_ (an oxyneurine),
which possesses considerable toxicity, and of which 0.00005 Gm.
suffices to kill a frog; _neurine_ (trimethylethylammonium hydroxide);
_choline_ (trimethyloxyethylammonium hydroxide); _mycetomuscarine_;
_anhydromuscarine_ (an oxyneurine); and a whole series of various
betaines.

=Symptomology.=--It is quite natural to divide this symptomology into
three different periods; that of incubation, that of manifestation of
symptoms, and that of termination.

The duration of the first period, that of incubation, is exceedingly
variable; it very rarely lasts more than forty-eight hours, and becomes
general only a few hours after absorption. Certain conditions influence
the duration; firstly the quantity of mushrooms ingested, then the
manner in which they were prepared; and, to some extent, the nature of
the organism, whether child or adult, healthy or in poor health.

When it is a question of the more particularly alkaloid-containing
mushrooms, especially when the poisoning is due to muscarine, the toxic
symptoms generally develop rapidly, the first symptoms appearing about
one hour after the ingestion of the mushrooms. On the other hand, if
the poisoning is due to one of the albuminoid group, and particularly
in the case of phallin, the period of incubation is longer, and may
last ten, twenty, thirty, or even forty-eight hours and more.

The symptoms begin with dizziness and an indefinable sensation of being
ill.

The second period is characterized chiefly by digestive and by nervous
derangements. The digestive derangements are evidenced by very violent
and painful vomiting, and diarrheas of choleraic or dysenteric
character. The nervous derangements vary according to whether they are
developed by an alkaloid, which causes delirium with hallucination, or
by albuminoids, which cause depression, ataxo-adynamia, and stupor,
these being particularly characteristic of the action of the toxic
albuminoids.

As for the period of termination, it results either in death or a cure.
If the poisoning is due to phallin, death appears to be an almost
inevitable consequence, as it occurs in 80 per cent. or more of the
cases. The poisoning by the alkaloids is less dangerous, and the cure,
when it does occur, is very rapid, almost immediate, in fact, while in
the case of the toxic albuminoids the cure is very slow, and attended
by relapses.

One characteristic of these toxalbumins is that they are apt to develop
specific antitoxalbumins. This fact has been verified not only in the
case of abrin, ricin, robin, and their analogues, but also in that
of the vegetable and animal diastases possessing toxic properties
even in the slightest degree only. These antibodies generally exhibit
their action _in vitro_. Thus antiricin exerts its antiagglutinative
action on the erythrocytes _in vitro_ in a saline medium in which the
erythrocytes cannot live.

Here, again, as in the case of the antitoxins, it must be admitted that
the antitoxalbumin possesses a specific affinity by virtue of which it
unites chemically with the toxalbumin to give rise to a new substance
which is devoid of toxicity.

The first antidiastase obtained by immunization methods, and according
to the mechanism we have already seen, was _antiemulsin_, obtained by
Hildebrandt.[62] This antiemulsin counteracts, both _in vivo_ and
_in vitro_, the specific action of emulsin. These studies have been
followed by a large number of scientists, particularly by Camus and
Gley,[63] Carnot, Mesnil,[64] and Charron and Levaditi,[65] in the case
of trypsin; and Sachs[66] in the case of animal pepsin. Gessard[67]
obtained a very active _antityrosinase_, and Mohl an _antiurease_.

 [62] HILDEBRANDT: Weiteres über hydrolyt. Fermente, etc. _Virch.
 Arch._, CXXXI, 1895, P. 5.

 [63] CAMUS and GLEY: _Compt. rend. de la Soc. de Biolog._, 1897.

 [64] MESNIL: Sur la digestion des actinies. _Annales de l'Institut
 Pasteur_, 1901.

 [65] CHARRIN and LEVADITI: _Compt. rend. de l'Académie dest Sciences_,
 1900.

 [66] SACHS: Ueber Antiseptika. _Zeitschr. f. Biolog._, 1901, XXVI.

 [67] GESSARD: _Annales de l'Institut Pasteur_, 1901, p. 609; _Comp.
 rend. de la Société de Biologie_, May, 1902.

The most important researches regarding this subject have been
published by Morgenroth, Briot,[68] and Korschum[69] on _antilab_ (or
_antirennet_). The researches of these authors have fully demonstrated
that there is considerable difference between the various rennets,
which had heretofore been confounded under one head; thus there is no
difference whatever between animal rennet and the rennet extracted
by Rosetti[70] from Cynara cardunculus (cardoon) so far as their
coagulant action on milk is concerned, yet each yields an antibody
which is strictly specific to itself. From a scientific point of
view we see, therefore, that the preparation of antidiastases permits
us to differentiate certain diastases that could otherwise not be
differentiated.

 [68] BRIOT: Thèse de Doctorat ès-Sciences, Paris, 1900.

 [69] KORSCHUM: _Zeitschr. f. physiolog. Chemie_, 1902, XXXI.

 [70] ROSETTI: _L'Orosi, giorn. di chemica, farmacia et scienza affini_,
 1898.


    III. ANIMAL TOXINS.

As we have shown at the beginning of this chapter, certain diastases,
and particularly those that are concerned with the digestive processes,
pepsin, trypsin, etc., and which are produced in abundance by the
entire living organism, possess quite clearly defined toxic properties,
and sometimes to even a considerable extent.[71]

 [71] GUSTAVE SAUX: De la toxicité des produits de la digestion
 peptique. _Thèse de doctorat_, Bordeaux, 1902.

Hemialbumose, from which peptones are formed, is itself a dangerous
toxin. It is generally believed that the toxic action of the peptones
and of the products of digestion of the albuminoids is due not to
the peptone itself, but to the more advanced products of digestion,
alkaloidal products unquestionably closely allied to the ptomaines.

Nevertheless, the true peptones behave just like true poisons, when
they are introduced hypodermically into the blood.[72]

 [72] SCHMIDT: _Mühlheim, Arch. de physiol._, 1880.

Brieger has made us acquainted with a non-proteid substance, under
the name of "peptotoxin," which is met with at the beginning of the
putrefaction of albuminoids. This toxin, which is not a protein, is
nothing else but a ptomaine. It is not altered by heat, and possesses a
very high toxicity. Brieger claims that it is a hydroxylized derivative
of an aromatic amide.[73]

 [73] BRIEGER: _Berichte d. D. chem. Gesellsch._, XIX, p. 3120; and
 _Verhandl. d. Congress f. innere Med._, II, p. 277.

Besides these facts, experiment has shown that the leucocytes, or white
corpuscles, the defensive rôle of which we have noted in phagocytosis,
owe their properties to the ferments which they secrete, and
particularly to some of the digestive ferments. These white corpuscles
are very rich in ferments of all kinds. Rossbach found in them amylase;
Achalme found lipase, casease, and trypsin; and the study of immunity
has brought to light a series of other ferments, the alexins or cytases
(microcytase and macrocytase), which have an exceedingly important rôle
to play.

It may easily be conceived that under certain circumstances a part or
the whole of these ferments can pass into the blood of the fluids of
the body, when they give rise to serious disturbances in certain cases,
or confer immunity in others.

It is thus that, according to Gautier, the rise of temperature which
characterizes fever is a consequence of the abnormal transudation of
these normal ferments into the blood, and their transmission by the
general circulation to the nervous centers.

However, it is not only in the leucocytes that we meet with these
toxic digestive ferments; it appears quite probable, and has even been
partially demonstrated, that they occur in a large number of other
cellular elements.

It is not necessary here to dwell upon the formation of the antibodies
of this group of active substances. The animal toxins are animal
diastases, and we have seen in the preceding paragraph that these
substances yield specific antibodies with great facility. For the rest,
we will dwell more fully on these antibodies of the animal toxins in
another volume of this collection, specially devoted to the study of
these substances, and entitled "_Les Serums Immunisants_," to which we
refer the reader who is desirous of obtaining more complete details
than he can obtain in the present volume.

=Alimentary Intoxications.=--What we have already stated permits
us to understand the phenomena of indigestion and botulism. The
toxic substances form within the digestive tract when the nervous
conditions modify the composition of the gastric juice, and arrest
the flow of hydrochloric acid, the presence of which normally checks
the development of the microbial flora, so rich within the stomach.
The result is the production, within the organism, of all kinds of
dangerous toxins. The same thing happens when the liver does not
functionate normally, and this, affords us a knowledge of the mechanism
by which foods that are most wholesome may become toxic by reason of
poor digestion or poor assimilation.

The absorption of spoiled viands may, _a fortiori_, produce serious
results. The alteration may be due not only to a bacterial infection,
as in tainted meat, but it has also been proved that the flesh of an
animal that has died of terror or madness may be very dangerous as a
food, even after cooking, because, although there are toxins which are
destroyed by a sufficient heat, there are ptomaines and certain toxins
that resist destruction under these conditions.[74]

 [74] POLLIN and LABIT: _Examens des aliments suspects_, Masson,
 publisher.

The use of preserved but spoiled beef, preserved ham or birds, sausages
frequently, and pieces of pork tainted by sausage poison, gives rise to
a succession of toxic symptoms the principal ones of which are dryness,
constriction of the pharynx, bilious vomiting, diarrhea, dyspnea with
pulmonary edema, etc. Fish and eggs are foods quite frequently capable
of developing serious results; the same is the case with molluscs,
mussels, oysters, lobsters, and snails. Lastly, moldy bread, spoiled
cheese, putrid water, and spoiled vegetables themselves, are proper
agents for determining attacks of botulic poisoning.

We have seen, at the beginning of this volume, that putrid meats
contain ptomaines, which are among the most toxic alkaloidal bases. We
have shown that Brieger has isolated from them neuridine, putrescine,
muscarine, and guanidine; that Nencki has isolated hydrocollidine;
and that Gautier and Etard have obtained from them parvoline--only to
mention a few of them.

Lastly, there may develop within the gastrointestinal tract dangerous
putrefactions, the products of which may enter the veins and arteries
from the ileum (a portion of the small intestine) and be distributed
throughout the organism. Although such poisonings occur, they do not
immediately follow the ingestion of the spoiled or toxic foods, but
they are always preceded by a period of incubation varying from several
hours to several days.

These alimentary poisonings are recognized by a great physical
depression, accompanied by vomiting and paralysis of the lower
extremities, sweats, and diarrheas. In some cases there occur cutaneous
eruptions; and when death happens, this occurs only several days later,
and generally without being preceded by any great pain.

=Urinary Toxins.=--As we have already remarked several times, it is by
the renal way that the organism voids its principal waste products.

We have seen also that it is by the kidneys that the toxins are
eliminated in all pathological conditions. As a general rule, the
urines are always more or less toxic. This toxicity of the urines
must be attributed in the first place to the crystallizable organic
principles (ptomaines and leucomaines[75]) which they contain;
secondly, to the non-crystallizable[76] extractive matters not so well
known; and lastly, to the saline substances, among which the potassium
salts are the most active. We find these mineral salts particularly
abundant under normal conditions in the urines of the herbivora.
According to Bouchard, 0.18 Gm. of potassium chloride are sufficient
to prove fatal to 1000 Gm. of living organism; a man excretes on the
average 2.5 Gm. of this salt, and a rabbit excretes about double this
quantity, weight for weight.

 [75] ADDUCO: _Arch. Ital. de biolog._, 1891.

 [76] POUCHET: _Thèse de Doctorat en Médecine_, Paris, 1878.

A very large number of hypotheses have been advanced regarding the
toxicity of the urines. Wilson considers the urea as being responsible
for it; Stadthagen[77] believes it to be due to the potassium salts,
etc. Bouchard[78] was the first to recognize that the toxicity of the
urines is due to a number of causes. We will not dwell further on these
active principles which, in the last analysis, are no other than
those that form in the various portions of the organism, and which are
eliminated by the urine.

 [77] STADTHAGEN: _Zeitschr. f. Klin. Med._, XV.

 [78] BOUCHARD: _Leçons sur les Autointoxications_.

It is self-evident, and it has already been shown, that the toxicity
of the urines varies greatly according to the malady, in consequence
of the elimination of toxins by the urines. According to Bouchard, in
infectious maladies the urines are twelve times more highly charged
with toxins than is blood serum. Moreover, the toxicity of the urines
is considerably augmented the moment there is the least febrile
condition, no matter what the cause is.[79]

 [79] Regarding this point see the excellent work by A. CHARRIN:
 _Poisons de l'Organism_. Masson, publ.

Even in the normal condition, the urinary toxicity varies greatly; and
this is easily conceived since the physiological phenomena that control
this secretion undergo incessant rise and fall. Thus, for example, the
urines eliminated during sleep are less active than those produced
during waking, because during sleep the elimination of cellular poisons
is at a minimum. Exercise, walking, physical and intellectual labor,
exert their portion of influence on these oscillations of toxicity;
and this variation of toxicity is due not to any one variation in the
mineral extractive matters, but rather more or less to the organic
toxic products. We will not dwell further on this subject, but will
simply refer to the work by Charrin, already mentioned, for all
supplementary details.

=Autointoxications.=[80]--The cells of the organism having, as a whole,
a life very much like that of the microbes, it is quite natural that
among the excreted products of the living tissues there should be found
the same substances formed as a result of the anaerobic fermentation of
albuminoids. Experiment has demonstrated that this is so, and Armand
Gautier has irrefutably proven the existence of these principles.[81]
Bouchard was the first to demonstrate the toxic nature of muscle
extract,[82] and Roger[83] established the fact that the toxicity of
this extract is due to ferment-toxins; it has since been recognized
that after death these toxins accumulate in the muscles.

 [80] CH. BOUCHARD: _Des Autointoxications_. Paris, 1887.

 [81] _Bull. Acad, de Médecine_ (2), X, p. 947, and XX, p. 115.

 [82] BOUCHARD: _Leçons sur les Autointoxications_, Paris, 1887.

 [83] ROGER: Toxicité des Extraits des Tissus Normaux. _Soc. de
 Biolog._, 1891, p. 728.

The extract of kidney made rapidly by cold process by triturating the
washed kidney with glycerin, and precipitating the glycerinic solution
with alcohol, contains toxic ferments to which the name "_hystozymes_"
has been given.[84] These ferments split up hippuric acid into benzoic
acid and glycocoll. Lépine has likewise discovered in the kidney a
very toxic pyrogenic substance.[85] Roger has given us evidence of the
toxic properties of the liver, washed and pulped, and then sterilized
by filtration through a porous diaphragm. This scientist has shown that
the toxic properties are due to albuminoids, which lose their activity
when heated to 100°C.[86]

 [84] It is well to recall here that the kidneys contain both reducing
 and oxidizing ferments, as has been demonstrated by de Rey-Pailhade,
 and later by Abelous and Gérard.

 [85] LÉPINE: _Compt. rend. de l'Acad. des Sciences_, May 13, 1889;
 _Soc. de Biol._, 1891, p. 724.

 [86] ROGER: _Compt. rend. Soc. Biol._, 1891, p. 727.

It must be remarked that the organs we have studied are essentially
reducers, and that the more powerful reducers yield the most toxic
extracts. We find here a confirmation of Armand Gautier's views
regarding the anaerobic origin of the toxic substances formed within
the organism.[87]

 [87] POZZI-ESCOT: _Compt. rend. de l'Acad. de Médecine_ (3), XLVII, p.
 400. See also POZZI-ESCOT: _Etat actuel de nos Connaissances sur les
 Oxydases et les Réductases_. Dunod, publ., Paris. 1902.

Blood serum precipitated by alcohol affords products which possess very
marked toxic power. It would appear that the toxic products we speak
of here are thermogenic diastatic substances derived from the white
blood corpuscles. In certain diseases the blood serum may acquire a
high degree of toxicity. We will recur again presently to this property
as a normal characteristic of the blood of various animal species, and
will study it in greater detail in a future volume of this collection,
devoted to the immunizing active principles.

=Glandular Secretions.=--On studying the venoms we will see that a
certain number of these products are the result of glandular secretion.
This is a general property of the glands; and it was Brown-Sequard who
first drew attention to the rôle played by these glands, and to the
importance of the products that they throw into the blood.[88]

 [88] _Compt. rend. de l'Acad. des Sciences_, CXIV, pp. 1237, 1318,
 1399, and 1534; CXV, p. 375; and CXVI, p. 856.

P. Noel showed later that the testicular juice possesses a high degree
of activity, which he attributed to an oxidizing ferment, and which we
have already mentioned, _spermine_.

The greater number of the other glands contain proteid matters and
various peptones, more or less toxic, with amides and alkaloids.

Particular mention must be made of the thyroid gland, the secretions
of which exercise a powerful action on the nervous centers and on
nutrition.[89] It appears reasonable to attribute to the secretions
of this gland a very powerful antitoxic action, and the first proof
of this fact is that the organisms deprived of this gland become the
seat of serious derangements; the urines of such organisms become
particularly toxic, while, on the other hand, the hypodermic injections
of the aqueous extract of the gland, when the derangements spoken of
exist, cause the immediate disappearance of the derangements caused by
the excision of the gland.[90]

 [89] LAULANIÉ: _Compt. rend. Soc. de Biol._, 1894, p. 187.

 [90] GLEY: _Compt. rend. Soc. de Biol._, 1891, p. 250.

Attempts have been made to isolate the active principle of the glands.
Notkine isolated a _tyroproteid_,[91] which is not sensibly toxic to
animals who still retain the gland, but which becomes toxic when the
gland is excised. It seems probable, however, that this product is not
the principal agent of the thyroid gland.

 [91] _Semaine Médicale_, Apr. 3, 1895, p. 138.

From the researches of Schaeffer, Roos, and Sigmund Fraenkel[92] it
results that the active principle of the gland is not a toxin, but a
purely chemical substance, a true leucomaine, which has received the
name _thyroantitoxin_.

 [92] _Wiener Med. Blätter_, No. 48; and _Gesellsch. d. Aerzte in Wien_,
 Nov. 22, 1895.

On the other hand, Baumann quite recently extracted from the thyroid
gland an iodized substance, which he named _thyroiodine_.[93]

 [93] _Zeitschr. f. Physiol. Chem._, XXI, pp. 319 and 481; and XXII, p.
 1. ARMAND GAUTIER: Chimie Biologique, 2d edit., pp. 330-332. Masson,
 publ.

The suprarenal capsules also possess properties that have often
attracted the attention of physiologists during the last few years.
They are considered as being, just like the thyroid gland, producers
of antitoxins; they destroy, or seem to destroy, toxins that are
artificially introduced into the circulation.

Albanèse[94] maintains that the function of the suprarenal capsules
is to neutralize neurine, the toxic product of the disassimilation of
the nervous system; this view, however, is opposed by Boinet[95] and
Langlois.[96] On the contrary, it has been definitely proven that the
suprarenal glands exert a specific action on the poisons of muscular
origin. Abelous and Langlois[97] have in fact demonstrated that the
alcoholic extract of the muscle of a decapsulated animal has the same
properties as the extract of tetanized muscle; the decapsulated animal
gives ergographic tracings analogous to those afforded by tetanized
animals. The removal of the suprarenal capsule from an animal brings
results, hence, analogous to those of fatigue--that is to say, that
the toxic substances which accumulate as a result of the decapsulation
resemble those that result from muscular exertion. The suprarenal
capsules exert their action furthermore on other toxic products as
well, as Guieysse[98] has shown, and particularly on the exogenous
poisons. In conclusion, it may be said that the matter concerns a
most important rôle, and we cannot do better in this respect than
to refer the reader to the memoir presented by Sergent and Bernard
to the Académie de Médecine in 1902 and entitled _l'Insuffisance
Surrénale_.[99]

 [94] ALBANÈSE: Recherches sur les fonctions des capsules surrénales.
 _Arch. Italiennes Biol._, 1892.

 [95] BOINET: _Compt. rend. Soc. de Biol._, Mch. 1896.

 [96] See _Compt. rend. de Biol. et Arch. Physiologie_, 1891-1897.

 [97] LANGLOIS: Thèse de doctorat en Méd., Paris, 1897.

 [98] GUIEYSSE: _Les capsules surrénales du cobaye_, Thèse, Paris, 1901.

 [99] Encyclopédie Léauté, CCCXIV, Masson, publ., Paris, 1904.




CHAPTER IV.

THE MICROBIAL TOXINS.


There is but one way of characterizing the toxic poisons secreted
by microbes, and that is to apply to them the name of the microbes
generating them; thus the soluble and toxic poison of the tetanus
bacilli has received the name _tetanus toxin_.

In toxic microbial cultures it is necessary to distinguish the toxins
proper from the toxic alkaloids (ptomaines) which generally accompany
them; this is easily accomplished by evaporating the solution in a
vacuum at about 30°C., and then treating with alcohol and ether, in
which the alkaloids are soluble, while the true toxins are insoluble.
By fractional precipitation with alcohol it is easy to isolate the
peptones and true toxins.

The microbial toxins possess two essential properties; one the pyogenic
property, thanks to which the toxins first attract, then destroy the
white blood corpuscles or leucocytes, and transform them into pus, and
the other the pyretogenic property, which appears to belong only quite
indirectly to the pyogenic substance. The toxins in general retard the
heart action.

We will not speak of the distinctions it has been sought to establish
between the substances which possess these different properties, but
will at once take up the discussion of several of the microbial toxins.

=Anthrax Toxin=[100] (from Bacillus Anthracis).--We will describe the
preparation of this toxin as a type.

 [100] ARLOING, CORNEVIN, THOMAS: _Le Charbon Symptomatique_, 1st edit.,
 Paris; and LE DANTEC: _La Bactéridie du Charbon_, Masson, publ.;
 STRAUS: _Le Charbon des Animaux et de l'Homme_, Paris, 1887.

The cultures of the bacillus are made in Liebig's bouillon, to which
has been added 0.1% of fibrin, the whole being carefully sterilized
for a long time at 110° C. The cultures medium is inoculated with a
drop of blood taken from the heart or spleen of an animal that has
died of anthrax. At the end of a week, the culture is filtered, and
the filtrate acidulated with a little acetic acid and precipitated
by adding powdered ammonium sulphate. The flocculent precipitate
is collected, washed, dissolved in distilled water, and dialyzed.
The dialyzed solution is concentrated in vacuo at 40-45° C., and
precipitated by adding to it alcohol. The precipitate formed is then
collected and dried.[101]

 [101] HANKIN: _British Medical Journal_, Oct. 12, 1889, and July 12,
 1890.

In this manner there is obtained a grayish-white substance which is
soluble in water, and which is fatal in large doses, but which, given
in repeated small doses, confers immunity against anthrax.

According to Hankin, it seems that the toxic property of this toxin is
due to an albumose.

Marchoux[102] has been able to confer immunity upon sheep by injecting
first small quantities of the filtered culture of the anthrax bacilli,
and then the virulent anthrax itself.

 [102] _Annal. Instit. Pasteur_, IX, p. 785.

The animals thus rendered immune yield a serum which may be used as a
vaccin against anthrax, and which even possesses curative properties
under certain conditions.

In every case the acquired immunity is only temporary. We will recall
to recollection the method employed by Pasteur for vaccinating against
anthrax, using attenuated cultures, a method which is practiced daily
at the present time.[103]

 [103] CHAMBERLAND: _Le Charbon et la Vaccination Charbonneuse_, Paris,
 1887. PETERMANN: _Annal. Instit. Pasteur_, VI, p. 32.

From the cultures of symptomatic anthrax (Bacillus Chauvæ) Chauvée
extracted a very active toxin which can withstand without impairment
a temperature of 110°C.[104] Roux[105] has shown that the serum of
animals that have succumbed to the symptomatic anthrax is capable
of vaccinating against this disease; we have here a new proof that
the antitoxin is in fact a product of the defense of the cells of
the organism, and the author mentioned has been able to vaccinate
guinea-pigs by injecting into the peritoneum culture bouillon
sterilized by heating to 115° C. or by filtering through porcelain.

 [104] DEUTSCHMANN: _Annal. Instit. Pasteur_, VIII, p. 403.

 [105] _Annal. Inst. Pasteur_, Feb. 1888.

=Tubercular Toxin.=--The culture bouillons of Koch's bacillus contain
one or more active substances which constitute, and which is at the
present designated as, tuberculin.[106] Koch's therapeutic tuberculin
is obtained by evaporating to one-tenth its volume a culture bouillon
of Koch's tubercle bacilli prepared from a 4-per cent. glycerinic
mutton bouillon, and filtering through porcelain. By fractional
precipitation it is possible to obtain from the crude tuberculin so
prepared a product which is considered as pure tuberculin, and which
possesses considerable activity.

 [106] AUCLAIR: _Thèse de doctorat_, Paris, 1897; and _Arch. de
 Médecine_, exp. 1898.

Prolonged boiling on the water-bath completely destroys the activity
of this tuberculin, which moreover hardly ever keeps longer than three
weeks. It has been found possible to preserve it for an indefinite
period, however, by adding to it 30 to 40 per cent. of glycerin. It
possesses all the general reactions of albuminoids.

Tuberculin is not toxic in the proper sense of the word. Injected in
small quantities into the healthy human being[107] and into healthy
animals, it exerts no effect; on the other hand, however, in tubercular
organisms, even in incipient stages of the disease, even where it is
almost impossible to make a clinical diagnosis, the injection of very
small quantities develops a lively and characteristic reaction.[108]

 [107] KOCH: _Deutsch. Med. Woch._, Nov. 13, 1890-1897, No. 14, p. 209.

 [108] _Annal. de l'Instit. Pasteur_, V, p. 191; _Arch. de la Soc. Biol.
 de Saint-Pétersbourg_, I, p. 213.

Grasset and Vedel consider the tuberculin as an excellent means of
diagnosing tuberculosis in man, but in such a case it is necessary
to operate with the greatest caution, with very small quantities of
the tuberculin, and to feel, in some sort, the sensitiveness of the
patient, particularly in the case of children.

It is chiefly for the diagnosis of tuberculosis in cattle, however,
that tuberculin is valuable. Thanks to Nocard, the procedure has to-day
become a common practice. The injection of a fairly large dose, 0.3
to 0.4 Gm., according to the size of the animal, causes, in about ten
hours or so, if the animal is tuberculous, a strong febrile reaction
with an elevation of temperature of 1.5 to 3° C., whereas if the animal
is not tuberculous no such reaction takes place.

Cases in which tuberculosis is far advanced, and in which the organism
is impregnated with tuberculin, do not react after the injection of
tuberculin.[109]

 [109] NOCARD and LECLAINCHE: _Les Maladies Microbiennes des Animaux_.

Tuberculin does not confer immunity, and the bacillus retains all its
virulence, even in injected tissues; nevertheless, the return to health
of animals in which injections have been recently made may be due to
the action of large doses of the serum; and on the other hand the
tuberculin, in large quantities, may render the location unsuitable for
the development of the tubercle bacilli.

=Diphtheria Toxin.=--The most characteristic property of the diphtheria
bacillus is the production, in culture media, of a special toxic
substance which has been named _diphtheritic toxin_; this name,
however, has come to be also extended to a liquid in which the bacilli
have lived, and which has been sterilized by filtration or by any other
suitable process.

Roux and Yersin[110] were the first to affirm that diphtheria is an
autointoxication caused by a very active poison formed by the microbe
in the restricted locality where it develops. In order to obtain this
toxin[111] a culture of the bacillus is first made in a mutton bouillon
made strongly alkaline with sodium carbonate (10 grams per liter), and
with the addition of 2 per cent. of peptone. At the end of about one
month, the culture being kept at about 37° C., the liquid is filtered
through porcelain. It is indispensable to employ a very virulent
bacillus; it is hence frequently advantageous to increase the virulence
and toxigenic power of the bacilli it is desired to use.

 [110] _Annal. de l'Instit. Pasteur_, II, p. 632, and VIII, p. 611.

 [111] See SPRONK: _Annal. de l'Instit. Pasteur_, IX, p. 785; _Ibid._,
 X, p. 333; MARTIN, _Ibid._, XII, p. 26; SPRONK, _Ibid._, XII, p. 711.

The toxic liquid obtained is exceedingly powerful: 0.1 Cc. kills a
rabbit in forty-eight hours. This toxin is very sensitive to the
effects of heat. When heated to 65° C. it loses almost all its
toxicity; at 70º C. it becomes innocuous; and it only requires to be
heated to 100° C. for fifteen minutes in order to lose all immediate
activity even in large doses. Nevertheless toxins thus weakened are
capable of proving fatal to an animal even after five or six months.

Light, oxygen, ozone and all oxidizers destroy the active principle of
the diphtheria toxin, which is, moreover, rendered almost inactive by
organic acids.

This toxin is capable of diffusing through animal membranes, a fact
that is in agreement with the toxic effect seen in a subject attacked
with diphtheria, and due to the toxin passing through the mucosa. In
spite of this property, however, the diphtheritic poison may be taken
into the stomach without any pernicious results.

Roux and Yersin have shown that, like all the diastases, it may be
precipitated from its solutions by the development, within these,
of certain precipitates, particularly calcium phosphate. It is
precipitated from its solutions by alcohol, as has been observed
also in the case of diastatic solutions. All the toxic substance
is contained in the albuminous precipitate thus obtained; but the
prolonged action of alcohol, or repeated successive precipitations,
alter it finally. Diphtheria toxin is likewise precipitated by the
reagents for albumoses, particularly sodium sulphate in saturated
solution. This procedure has been utilized by Brieger and Fraenkel
for preparing the pure toxin, which they obtained in the form of very
light, brilliant white, amorphous flocks, affording all the principal
reactions of the soluble albumoses (biuret, xanthoproteic, Millon's),
and which they characterized as a toxalbumin.

On injecting into healthy animals this diphtheria toxin attenuated
by sufficiently heating at 70° C, employing at first small doses,
and gradually increasing, it is possible to immunize them against
diphtheria, as was first demonstrated by Carl Fraenkel.

Roux and Martin, who have specially studied this procedure,[112] have
shown that a horse may be easily immunized by injecting into the animal
the toxin diluted with a third of its volume of Gram's iodine solution,
and in successively increasing doses. The initial dose is 0.25 Cc.;
then, after two days, 0.5 Cc. of the same toxin is injected, and in
like manner the dose is increased up to the eighteenth day, when the
pure toxin is injected, at first in small doses, which are gradually
increased so that at the end of two or three months injections of 80
Cc. of the pure toxin may be given without danger; the animal is then
completely immunized.

 [112] Contribution à l'Étude de la Diphtérie. _Annal. de l'Instit.
 Pasteur_, VIII, p. 609; _Ibid._, p. 640.

The serum of an animal rendered immune in this manner contains a
diphtheria antitoxin which possesses high power. A guinea-pig which
has received an injection of 0.01 Cc. of the antitoxin is perfectly
capable of withstanding a lethal dose of 0.5 Cc. of the toxin. The
antidiphtheria serum thus obtained, and in almost limitless quantities,
from an immunized animal, is capable of saturating the therapeutic
diphtheritic toxin, and has to-day taken rank in therapeutics as the
most efficacious remedy in diphtheria. Injected in varying doses, it
confers a temporary but immediate immunity.

Nevertheless antidiphtheria serum must not be considered as an
antidote; and in pathological diphtheria, the more serum is required
the later it is used.[113] In certain cases, if employed too late, it
may prove ineffective.

 [113] BAYEUX: _Thèse de Doctorat_, Paris, 1899.

The preventive action of the serum is remarkable. In 10 000 inoculated
cases Behring and Ehrlich have had but 10 cases of diphtheria, and
these were, moreover, of a benign character. The duration of the
immunizing action appears to be from three weeks to two months.

This diphtheria antitoxin was first prepared by Guérin and Macé[114] by
adding to the antidiphtheria serum a large volume of alcohol, washing
the precipitate, and drying it in a vacuum. It is soluble in water, and
loses its activity when heated to 65° C. Wassermann[115] has proposed
to extract it from the milk of immunized animals, by first coagulating
the milk by rennet in the presence of sodium chloride, filtering, and
removing the fat from the clear liquid by means of chloroform. After
decanting, the clear solution obtained is precipitated by adding to it
30 to 33 per cent. of ammonium sulphate. The precipitate is dried in a
vacuum on a polished porcelain slab after having first been strongly
expressed. It is then dissolved in water.[116]

 [114] _Compt. rend. de l'Acad. des Sc._, Apr. 5, 1895.

 [115] _Zeitschr. für Hygiene_, XVIII, p. 235.

 [116] ROUX and MARTIN: Contribution à l'Étude de la Diphtérie. _Annal.
 de l'Instit. Pasteur_, VIII, p. 512.

=Tetanus Toxin.=--The fact that the tetanus bacillus never penetrates
to the interior of the organism enabled us long ago to foretell that
it secretes a very powerful toxin capable of dialyzing and diffusing
through the economy. Kuno Faber was the first to fully recognize the
fact that the culture bouillon of this bacillus, fully sterilized by
filtration through porcelain, possesses an exceedingly high toxicity,
and exerts a toxic effect on 50 000 000 times its own weight of living
organism. Brieger had previously, however, extracted three ptomaines
from the cultures of the bacillus--_tetanin_, _tetanotoxin_, and
_spasmotoxin_.[117] In order to obtain a highly active liquid, the same
culture medium is inoculated several times in succession, but filtering
each time before the new inoculation; the microbes greatly increase in
number after each fresh inoculation, and the toxic substance developed
by them accumulates.[118]

 [117] Die Pathogenese des Tetanus. _Berlin. Klin. Wochenschr._, 1890,
 No. 31.

 [118] NAILLARD: _Compt. rend. de l'Acad. des Sciences_, CXX, p. 1181.

Experiment has shown that the culture bouillon thus obtained contains
two kinds of toxic substances[119]--highly toxic alkaloidal bases
(ptomaines, tetanin, tetanotoxin, etc.), and a true toxin, possessing
diastatic properties, and of almost incredible toxic power.

 [119] _Annal. Instit. Pasteur_, V, 15.

This toxin had already been isolated by Kitasato. It is a toxalbumin,
and is very sensitive to the action of heat. A temperature of 65° C.,
maintained for 30 minutes, renders it quite inactive; and it becomes
oxidized and is destroyed by the action of the air in the presence of
light.

Brieger and Boer,[120] by precipitating with zinc chloride the filtered
culture bouillon, obtained a pure, amorphous tetanus toxin, which they
also considered as a toxalbumin, and which possesses exceedingly toxic
properties.

 [120] _Deutsche Med. Wochenschr._, No. 49, Dec. 3, 1896.

If a precipitate be caused to form in these toxic solutions, as, for
instance, a precipitate of calcium phosphate, this carries down with it
all the toxin present in the liquid. 0.0005 Gm. of this precipitate is
surely fatal to a guinea-pig.

Dozon and Cournemont have observed that even in doses of 300 to 400 Gm.
of the filtered culture liquid, this toxin is not immediately toxic
to a horse, but kills the animal only after a period of incubation
of at least twenty-four hours. The blood of such an animal, however,
is immediately and directly fatal to animals into which it is
injected.[121]

 [121] _Compt. rend. Soc. Biol._, 1893, p. 294; _Ibid._, 1894, p. 878.

Experiment has shown that animals that have been cured of tetanus
possess no immunity whatever against tetanus; nevertheless Behring and
Kitasato[122] first, and Wassermann and Kitasato later on, succeeded
in preparing a _tetanus antitoxin_. To obtain this, the immunization
of the animal, horse or cow, is effected by injecting increasing
quantities of the toxin, more or less attenuated by mixing it with
Gramm's iodine solution; the immunization is easily and rapidly
accomplished by the process devised by Roux and Vaillard.[123]

 [122] _Deutsch. Med. Wochenschr._, 1890.

 [123] _Annal. Instit. Pasteur_, VII, p. 64.

The immunized animals yield a serum which, mixed with tetanus cultures,
renders these innocuous, and which enjoys an antitoxic power that
borders on the marvelous.[124] A quintillionth of a cubic centimeter
of the serum per gramme weight of a live mouse suffices to protect the
animal from an otherwise fatal quantity of tetanus toxin.[125]

 [124] NOCARD: _Bull. de l'Acad. de Médecine_, Oct. 22, 1895.

 [125] NAILLARD: _Compt. rend. de l'Acad. de Sciences_, CXX, p. 1181.

This serum is nevertheless powerless to preserve man in cases of acute
tetanus; it confers an immediate, but only transitory, immunity.

As to its mode of action, it appears to cause a permanent condition
of excitation or of nutritive reaction of the cells, which makes
these resistant to the poison. As in the case of the other toxins,
the quantity of antitoxin necessary to protect an organism is so much
greater the later the treatment is applied.

=Mallein (Toxin of Glanders).=--Among the soluble products secreted
in the culture media by the glanders bacilli, there are found true
toxins to which are ascribed certain symptoms of glanders infection.
These toxins have been isolated and designated by the name _mallein_.
First prepared by Helman and Kalmino, mallein was later on specially
studied by Roux and Nocard, and, in consequence of the researches of
the last-mentioned scientist, it has acquired great importance.[126] It
is obtained by sterilizing at 110° C. cultures of the glanders bacillus
made with mutton bouillon with the addition of salt, glycerin, and
peptones. To isolate the toxin the culture bouillon is first sterilized
by heating for half an hour in an autoclave at 100° C. It is then
filtered, concentrated to one-tenth its volume on a water-bath, and
filtered through a Chardin filter. The mallein is thus obtained in the
form of a brown syrupy liquid containing half its weight of glycerin.

 [126] NOCARD: _Les Maladies microbiennes des animaux_, Paris.

This solution keeps well when kept from air, light, and heat. In
practice it is employed in 10-per cent. solution in phenolated water
(5:1000). The mallein may be precipitated from the crude solution by
the addition of alcohol, as recommended by Foth. Foth's mallein occurs
as a white, light powder, very easily soluble in water.

Mallein enjoys a very important rôle in veterinary therapeutics, a rôle
analogous to that of tuberculin, permitting the diagnosis of incipient
glanders.[127]

 [127] STRAUSS: _Arch. de Médic. expériment_, 1886.

Experience has shown that in animals already attacked by glanders,
even if ever so slightly, the thermic reaction never fails when 0.25
Cc. of the mallein solution is injected. In healthy animals, however,
the injection of mallein, even in much larger quantities, causes no
apparent effect. In animals attacked by glanders the reaction attains
its maximum in twelve hours, and several days are required for the
temperature to return to normal.[128]

 [128] CADIOT and ROGER: _Compt. rend. Soc. Biol._, 1895, p. 770;
 WLADIMIROW: _Arch. des Sciences Biol. de St.-Pétersbourg_, IV, p. 30;
 BOURGES and MÉRY: _Soc. de Biol._, Feb. 5, 1878.

According to Nocard, mallein possesses no immunizing properties
whatever.[129]

 [129] GALTIER: _Compt. rend. de l'Acad. des Sciences_, XCII, p. 303;
 STRAUSS: _Arch. de Médic. expériment_, I, p. 489.

=Typhoid Toxin.=--This is obtained, like the other microbial toxins,
from a culture, prepared with more or less difficulty, from Eberth's
typhoid bacillus. This toxin, injected into guinea-pigs, develops in
them typhoid fever.

In the solution there occurs a ptomaine, which has been isolated by
Brieger, and which gives rise to almost all the phenomena of typhoid
fever; this ptomaine is called _typhotoxin_.[130]

 [130] BRIEGER: _Microbes, Ptomaïnes et Maladies_, Doin, publ., Paris,
 1887; LUFF: _Brit. Med. Journ._, 1889.

The same author, in collaboration with Fraenkel,[131] later on isolated
a toxalbumin from the culture bouillon of the typhoid bacillus.
Sanarelli[132] obtained an active toxin by macerating for several days
at 60° C. a month-old culture of the typhoid bacillus made with a 2-per
cent. glycerin-bouillon. Chantemesse has also published a process which
yields a highly virulent toxin.[133]

 [131] _Berlin. Klin. Wochenschr._, 1890.

 [132] _Annal. de l'Instit. Pasteur_, VIII, p. 103.

 [133] _Compt. rend. Soc. de Biol._, p. 232, Jan. 30, 1897. _Congrès
 d'Hygiène de Madrid_, 1898.

Chantemesse and Widal[134] have shown that on injecting into an
organism increasing quantities of the sterilized cultures of Eberth's
Bacillus, it is possible to fully immunize an animal against the
bacillus itself, and even also against the Bacillus coli communis. The
operation, however, is tedious and painful. The serum of immunized
animals possesses preventive and curative properties respecting the
effects of typhoid bacilli.

 [134] _Annal. l'Instit. Pasteur_, VI, p. 755; SANARELLI: _Ibid._, p.
 721.

A dose of the filtered culture, which is fatal to a guinea-pig,
becomes innocuous when mixed with 0.5 Cc. of the serum of a vaccinated
guinea-pig; 6 Cc. of the serum injected six hours after an injection
of the virulent culture, hence when this is in full action, suffice
to save the animal.[135] So far as the human being is concerned, the
results obtained have not been sufficiently satisfactory.

 [135] FUNCK: _La Sérothérapie de la Fièvre Typhoïde_, I, Brussels, 1896.

The culture bouillon of the Bacillus coli communis, which is closely
allied to Eberth's bacillus, also contains soluble toxic substances
which have been named coli-bacillus toxin. This substance, which is
produced only in small quantity by the microbe, is fatal only in very
large doses.

=Cholera Toxin.=--Very little is known regarding the toxic products of
the spirillium choleræ; nevertheless, the fact that typical cholera
exhibits every symptom of the action of a toxic agent demonstrates
quite clearly the elaboration of some toxic substance within the
cultures of this microbe.

Villiers[136] found in it a liquid ptomaine; Klebs[137] found another
and crystallizable ptomaine; while Pitai discovered in it a toxin
unalterable by heat, and which he considered as a toxopeptone.
According to Gamaleia[138] there is present a true toxin, alterable
by heat, and the reactions of which entitle it to be considered as a
nucleo-albumin; he has also found in it a toxic nuclein.

 [136] _Compt. rend. de l'Acad. des Sciences_, Jan. 12, 1885.

 [137] KLEBS: _Allgem. Wien. Med. Zeit._, 1887.

 [138] _Arch. de Méd. Expérim._, IV, p. 173.

These toxic substances are found, according to Gamaleia, Pfeiffer,
and Sanarelli,[139] confined during the life of the microbe within
its cellular envelope, and does not diffuse through this. Metchnikoff
and Roux are of the contrary opinion,[140] however, and they have
prepared a toxin almost insensitive to a temperature of 100° C., and
precipitable from its solutions by ammonium sulphate or strong alcohol;
the toxin is a toxalbumin. This toxin is quite toxic; one-third of a
cubic centimeter suffices to kill 100 Gm. of guinea-pig in 18 hours;
with larger doses, death is almost immediate.

 [139] _Annal. de l'Instit. Pasteur_, IX, p. 129.

 [140] _Ibid._, X, p. 257.

By immunizing guinea-pigs, rabbits, and horses with this cholera toxin,
Metchnikoff and Roux obtained a serum which is distinctly antitoxic for
rabbits. Nothing absolutely certain has been found as to its action on
man.[141]

 [141] HAFFKINE: _Compt. rend. de l'Acad. des Sciences_, 1892;
 METCHNIKOFF: _Annal. de l'Instit. Pasteur_, VII, p. 403; and ROUX:
 _Ibid._, X, p. 253.

       *       *       *       *       *

We will not dwell longer here on the toxins of microbial origin. It
appears evident, however, from what has been stated above, that the
great majority, if not all, of the virulent microbes manifest their
virulence by means of toxic secretions. Almost every one of these
toxins has been the subject of study. They would otherwise not have
interested us here, where our main object was but to dwell upon the
general properties.




CHAPTER V.

THE VENOMS.


=General Nature of Venoms.=--The venoms are more or less toxic products
secreted by certain reptiles, batrachians, and fish; by a large number
of invertebrates; by arachnids, apids, scorpionids, araneids, and a
large number of other insects.

The venoms are toxic principles very closely allied to the microbial
toxins; like the latter, they form two classes, the one alkaloidal,
the other proteid, possessing a true diastatic character. They
closely resemble the microbial toxins, moreover, by the fact that
they are capable of being transformed into vaccins by attenuation of
their virulence, by the action of heat or chemical reagents, and of
leading to habituation of use and the conference of immunity.[142]
Moreover, like the various viruses, the serum of immunized animals is
antivenomous, so that if injected into the veins or beneath the skin of
non-immunized animals, the serum confers upon them an immunity against
venom which lasts for some time.

 [142] _Annal. de l'Instit. Pasteur_, VIII, p. 281; _Journ. of
 Physiol._, VIII, p. 203; and _Soc. de Biol._, 1894, p. 111.

These venoms, like the microbial toxins, possess but slight toxicity
when absorbed via the stomach. Fraser, utilizing a method previously
advocated, succeeded, by following this method, in vaccinating against
serpent-venom by causing the absorption by animals of constantly
increasing doses of venom.

It was thus possible to make the animals withstand doses a thousand
times greater than the ordinary lethal dose; the blood and serums
of these animals at this point possessed immunizing properties, and
this property passed by heredity to the offspring, to which it is
transmitted by the blood itself, and by the milk during feeding.

Along with these resemblances between the venoms and toxins, attention
must be called to a very important difference. As we have already
seen, the action of the toxins on the organism is always preceded
by a certain period of incubation; the action of the venoms, on the
contrary, is almost instantaneous, and in this respect they behave like
chemical agents and alkaloidal toxins.

If the venoms are preserved in a moist condition, they change because
they undergo putrefaction, which is generally the case with all
diastatic substances, and particularly the toxins.

It is interesting to note that animals which have been bitten by a
venomous serpent, but which, for some reason or other, have not
succumbed to the venom, never recover their former condition; if they
were young, their functions cease to develop, and they droop; if they
are adults, their general condition remains that of stupefaction.

=Venomous Serpents.=--Among the venomous serpents,[143] the most
important as well as the most dangerous are the following: Cobra di
capello (Naja tripudians, the hooded cobra) and its analogues, the
black Naja, Naja hagé, etc.; the elops (coral serpent); the bungurus
of Bengal and Burmah; the Platycercus proteroglyphia, which is found
chiefly in the waters of the Indian Ocean; the crotalian solenoglyphs
of the two Americas, and among which in particular are the rattlesnake,
the fer-de-lance (the yellow viper) of Martinique; the surucucu of
Guiana; and the moccasins and copperheads of Texas and Florida. Lastly,
the entire group of viperian solenoglyphs, among which are the Echidnæ,
the bite of some of which, for instance the daboia or echidna, is
dreadful; the African vipers, among which may be mentioned the horned
viper, the bite of which will kill a camel; the springing viper of
Congo, and the rhinoceros-viper of Gabun; the European vipers, the most
dangerous of which is certainly the asp of France, which is exceedingly
numerous in certain regions.

 [143] CALMETTE: _Le Venin des Serpents_, Paris, 1896.

The effects of the bites of venomous serpents on man and animals
are generally well known to the public; it is well to recall them,
nevertheless. From the moment the bite has been inflicted, complete
symptoms of poisoning develop, attended by a condition of extreme and
increasing weakness, with vomiting, hemorrhage, and decomposition of
the blood. There are, besides, particular effects which vary with every
venom.

The following table by Calmette[144] gives the comparative toxicity
of various venoms, taking as the standard of comparison the quantity
sufficient to kill a rabbit in three or four hours:

  Naja tripudians           0.00047
  Naja hagé                 0.0003-0.0007
  Acanthophis antarctica    0.001
  Ceraste                   0.0017-0.0021
  Haplocephalus variegatus  0.0025
  Trigonocephalus           0.0025

 [144] CALMETTE: _Annal. Instit. Pasteur_, VIII, p. 276; IX, p. 229.

=Nature of Serpent-venoms.=--These venoms are homogeneous liquids,
somewhat more dense than water, in which they are soluble, slightly
colored green or yellow, transparent, and insoluble in alcohol; they
contain from 30 to 35 per cent. of solid matter. When fresh, they have
a slightly acid reaction. Towards chemical reagents, and particularly
acids, they behave like albuminoids; almost all the combinations they
afford with the various albuminoid reagents are active, despite their
insolubility. According to Gautier, they are decomposed by caustic
potash.

According to numerous researches, oxidizers like potassium
permanganate, the hypochlorites, hydrogen peroxide, and gold chloride
(in 1% solution) destroy the venoms; in certain cases when immediately
injected hypodermically in the poisoned region, these substances are
excellent antidotes _in vivo_.[145]

 [145] WINTER and BLYTH: _The Analyst_, 1877, p. 204; LACERDA: _Compt.
 rend. de l'Acad. des Sciences_, XCIII, p. 466; CALMETTE: _Annal.
 Instit. Pasteur_, VI, p. 175, and VIII, p. 278.

We shall not here enter upon a detailed study of the toxic albuminoid
principles of serpent-venoms; moreover, our knowledge is rather vague,
as it is, on a number of points. It will suffice us to know that, taken
altogether, the active albuminoids of these venoms are numerous, and
that each venom has its own particular active constituents, differing
according to the species and variety of the snake.

Each one of these substances acts more or less rapidly, and may be
associated with different principles which give rise to the variability
of the action of these toxic agents. Among these toxic albuminoids, the
most virulent appear to be true albumins and globulins, followed by
the nucleo-albumins, as we have already stated; there are also found
in venoms alkaloidal bases, but these principles are present only in
very slight quantity. These bases are but very slightly toxic compared
with the toxins that accompany them.

=Natural Immunity towards Serpent-venoms.=--Certain animals exhibit
a natural immunity toward snake-bites; among them are the snakes
themselves, the hog, the hedgehog, and the mongoos (an Egyptian rat);
the blood of these animals contains apparently an antitoxin.[146]

 [146] _Compt. rend. de l'Acad. des Sciences_, CXXI, p. 745; JACODOT:
 _Arch. de Médecine Navale_, VII, p. 390.

Fontana[147] had remarked that snakes were quite unaffected by the
bite of the viper, even when inoculated with the venom hypodermically.
Physalix and Bertrand[148] confirmed these statements, and showed
that the snake perfectly resisted quantities of viper-venom capable
of killing at least 20 guinea-pigs. According to these scientists,
this natural immunity is due to the existence in the blood of toxic
principles analogous to those of viper's venom--principles that exist
in the labial glands of the snake, and pass into the blood and the
fluids via the internal secretions. These writers, and also Calmette,
have shown that the blood of venomous serpents becomes antitoxic when
heated.

 [147] _Traité sur le Venin de la Vipère_, Florence, 1781.

 [148] _Archives de Physiologie_, 1894, p. 423.

It has been known for a long time that the hedgehog and the mongoos
eat certain venomous reptiles, and eagerly hunt for the vipers in
particular. When the hedgehog is bitten, which happens quite often
despite its dexterity, it resists the viper-venom quite well. Physalix
and Bertrand[149] have experimentally demonstrated that the hedgehog
withstands a dose of viper-venom capable of killing at least 40
guinea-pigs. Levin[150] has shown that young individuals are less
resistant, and it is concluded from this, and perhaps incorrectly so,
that the immunity of the hedgehog is naturally acquired, rather than
inherent. Bertrand and Physalix have nevertheless shown that on heating
the blood of the hedgehog to 88° C. it manifests an antitoxic power
toward serpent-venom _in vitro_.

 [149] _Bull. Muséum Histoire Naturelle_, I, p. 294; _Compt. rend. Soc.
 de Biol._, 1899. p. 77.

 [150] _Deutsche med. Woch._, 1898, p. 629.

=Artificial Immunity toward Serpent-venom.=--Immunity may be conferred
upon every individual by utilizing the method of habituation. This
fact was simultaneously elicited by Calmette, Bertrand, and Physalix.
To effect the immunity these scientists prepare an antivenomous serum
and inject it into animals, giving at first very small quantities of
the diluted venom, and gradually increasing the doses, and the periods
intervening between the injections. At the end of about two months of
this treatment, the immunity has reached its maximum. Certain rabbits,
thus slowly inoculated, have been able to withstand 0.04 Gm. of the
venom of the naja at a single injection; such rabbits then yield a
vaccinal serum.[151]

 [151] _Annal. de l'Instit. Pasteur_, 1895, p. 229; _Compt. rend. de
 l'Acad. des Sciences_, CXXII, p. 203.

At the Institut Pasteur at Lille there is prepared in this manner an
antivenomous serum from the horse; it is capable of acting upon 20 000
times its own weight. This has rendered great service in the treatment
of snake-bites, particularly in hot countries, where the accidents are
of daily occurrence. _In vitro_ it acts quite as well preventively
as therapeutically. It arrests the effects of the naja, the horned
ceraste, the trigonocephalus, the rattlesnake, and of almost every one
of the venomous serpents known.

The relatively considerable immunity possessed by certain
snake-charmers, and which passes for a magical gift, is due to nothing
else but a natural immunity, acquired perhaps by heredity, and it
always appears to follow as a result of a nonfatal snake-bite.

=Venoms of Batrachians and Saurians.=--We observe here a fundamental
difference between these poisons and those of snakes, as we shall see.
These latter, in fact, appear to owe all their toxicity to true toxins
which they contain, while the poisons of batrachians and saurians are
chiefly composed of alkaloidal bases.[152]

 [152] CLOEZ: _Compt. rend. de l'Acad. des Sciences_, XXXIV, p. 592.

The poison of toads and frogs (studied by Faust, Bertrand, and
Physalix) is chiefly secreted by the glands of the subcutaneous tissues
of these animals; it has but a very slight action on the unbroken skin,
but it rapidly inflames the nasal and buccal conjunctival mucosa. The
poison is a yellowish liquid, milky and viscid, with a waxy odor and
an insupportably bitter taste. It is strongly acid and caustic. When
dried, the poison yields to ether a fatty matter which, when absorbed
by an animal, plunges the latter into a coma that may end in death.

The residue insoluble in ether contains the non-toxic albuminoids, and
ptomaines, such as methylcarbylamine,[153] and isocyanacetic acid,
resulting from the decomposition of a lecithin that appears to be
soluble in ether.

 [153] _Ibid._, XCVIII, p. 538.

To obtain this venom, Physalix and Bertrand[154] skin the toads, first
chloroformed, and dry the skins in a vacuum over sulphuric acid; the
skins are then cleaned by treating with carbon disulphide to remove
fatty matters, and the toxic principles removed by means of 95-per
cent. alcohol; the poison so obtained, however, is impure. A better
procedure is to express the parotid glands which have been placed in
distilled water. Faust found in this venom a principle which he named
_bufonin_. Physalix and Bertrand isolated from it also a resinoid
substance soluble in alcohol and in a large excess of water; this
substance, which they named _bufotaline_, acts upon the heart. These
authors have also obtained another substance which has a paralyzing
action, and which they have named _bufotenin_.

 [154] _Ibid._, CXXVIII, pp. 45-48.

The poison of the common toad acts as a paralyzant upon the heart and
on the spinal marrow[155]; that of the common frog possesses similar
properties. The poison of the tritons is quite analogous to that of the
toads; it contains a lecithin hydrolyzable by water with the formation
of alanin, formic acid, and alpha-isocyanopropionic acid.

 [155] P. BERT: _Compt. rend. de la Soc. de Biologie_, 1885, p. 524.

Zalnosky[156] isolated from the glands of the skin of the salamander a
white, thick, bitter and alkaline liquid poison, containing a highly
poisonous alkaloid, _salamandrine_, or _samandarine_, which acts on
the brain, the medulla, and the spinal cord, and which has the formula
C{54}H{60}N{2}O{5}; it is a strong base and yields crystallizable
salts.

 [156] _Bull. Soc. Chim._ [2], VI, p. 344.

=Fish-poisons.=[157]--Very little accurate knowledge is extant
regarding these. Many fish are poisonous, and among them are the
synanceia, found in the Indian Ocean between the Netherland Isles and
New Caledonia; considerable numbers are found in the neighborhood of
the latter locality. These fish are provided with spiny rays which are
in direct communication with a poisonous system having its seat in the
dorsal fin. The prick of one of the spiny rays of this fish may under
certain circumstances result fatally, and in every case it causes a
rapid and painful gangrene.

 [157] BOFFORD: _Thèse de doctorat en Médecine--Les Poissons venimeux_,
 Paris, 1889; O. ARCOS: _Thèse de doctorat--Essais sur les accidents
 causés par les poissons venimeux_, Paris, 1887.

From the reservoir the poison is conducted to the sharp extremity of
the spines by a deep channel with which each spiny ray is provided; the
animal has 26 poison-sacs, two for each ray, and the sacs burst when
the corresponding sting is in any manner compressed.

The poison is an odorless liquid having a slight styptic or acidulous
taste, and exhibiting a bluish fluorescence; it rapidly becomes turbid.

The weevers, which are numerous on the shores of the Mediterranean Sea,
and which are also met with in the northeastern portion of the Atlantic
Ocean, are likewise very dangerous, which explains their popular names
"viper-weever," "spiderweever," etc. These fish are provided with a
double set of poisonous apparatus, the one opercular, which is the
more dangerous, and the other dorsal. The opercular spine has a double
channel in connection with a conical cavity hollowed out in the base of
the opercular bone. The bottom of this cavity is provided with special
cells which secrete the poison. The dorsal glands have a similar
structure.

The poison of the weever is a liquid, limpid when the fish is alive,
and turbid when dead; it has a slight bluish fluorescence, is neutral
in reaction, and is coagulated by acids and bases. It acts as a
paralyzant, its action being exerted on the medulla and spinal cord; it
retards the heart's action.

These examples will suffice; and we will not dilate further on this
subject, because, as already stated, but little is accurately known
regarding the subject, and what is known may be summed up as follows:
Fish-poisons always give rise to an intense pain, frequently with motor
paralysis, followed by paralysis of sensation; they affect the heart,
arresting it in diastole; and they are more dangerous to fish and
cold-blooded animals than to mammifers.

=Poisons of the Hymenoptera.=[158]--The poison system of the bee, and
of such insects as the wasps, bumblebees, etc., is known to consist of
a hollow sting consisting of two sharp needles communicating with two
poison-bearing glands, and forming a flexible tube. One of these glands
secretes an acid liquid (formic acid); the other secretes an alkaline
fluid.

 [158] PHILOUZE: Venin des Abeilles. _Annales de la Société Linn. du
 Maine-et-Loire_, IV.

The action of the bee-poison is very often benign, but there have been
cases where death followed the infliction of numerous stings.

Our information regarding the poison of the cantharides and flies is
very vague[159]; the same is true of the poisons of various arachnids,
acarides, and myriapoda. So far as spiders are concerned, it is known
that their poison is an oily liquid having an acid and bitter taste,
and containing a toxalbumin derived from the skin of the insect. The
bite of the ordinary spider occasions simply a slight local pain, with
redness; that of the large poisonous spider, however, may kill the
larger animals, and even man.

 [159] JOYEUX-LAFFRIÉE: _Thèse de doctorat en Médecine_, Paris, 1883; P.
 BERT: _Compt. rend. de la Soc. de Biol._, II [4], p. 136.

=Poison of Scorpions.=[160]--This poison is a colorless, acid liquid,
having a higher specific gravity than water, in which liquid it is
soluble. The famed legend of the suicide of scorpions is well known to
all. It is stated that when the insect finds itself in a position where
its death is inevitable, it stings itself, and dies from the effects of
its own poison. A simple method has even been described of bringing
this result about experimentally by surrounding the insect with a
circle of fire. Bounne, of Madras,[161] who has studied the procedure,
has demonstrated its entire falsity by showing, first of all, that the
insect dies from the effects of the excessive heat, and further, that
the poison of the scorpion is harmless to individuals of the species
that furnish it.

 [160] CALMETTE: _Annales de l'Instit. Pasteur_, X, p. 232.

 [161] _Proceedings of the Royal Society_, XLII, p. 17.

Metchinkoff[162] has confirmed these facts, and has moreover
demonstrated that the blood of the scorpion possesses an undoubted
antitoxic power against the poison of the insect.

 [162] METCHNIKOFF: _L'Immunité_, p. 344.

The poison of the scorpion serves it to kill the insects which are its
prey. Frogs and birds stung by the scorpion also generally die. A dose
of 0.0005 Gm. kills a guinea-pig in less than one hour; and according
to Calmette[163] less than 0.0005 will kill a white mouse in two hours.
Oxidizers destroy the toxicity of the poison. Guinea-pigs immunized
against the poison of the scorpion resist perfectly very large doses of
the poison.

 [163] CALMETTE: _Annal. de l'Instit. Pasteur_, X, p. 232.

=Poisonous Blood and Serums.=--It is an almost general fact that the
blood and blood serum of batrachians, eels, lampreys, snakes (even
non-poisonous ones), and hedgehogs are very poisonous. Mosso has
found in the blood serum of the lamprey a toxin possessing a strong
hemolytic power, and which he has named _ichthyotoxin_. O.5 Cc. of this
serum injected into a dog kills it in a few minutes. He also observed,
in 1888, that the blood of the eel, in like dose, kills a dog almost
immediately, and that the blood contains an ichthyotoxin analogous to
that of the lamprey.

This substance, which appears to be closely allied to the sero-albumin
of the blood, has a phosphorus-like, sharp, and burning taste. By
digestion it loses its toxicity, as well as by heating at 68° to 70° C.
It is easily obtained by precipitating with ammonium sulphate the serum
of eels, and dialyzing the precipitate dissolved in water. The power of
this substance is almost as great as that of the cobra poison, 0.002
Gm. being instantly fatal per kilo of dog.

The blood of snakes is likewise very toxic; the same is true of the
blood of the viper, as 0.02 Cc. will kill a guinea-pig in two hours.
All these bloods lose their toxicity when heated above 70° C. The serum
of the hedgehog is peculiar in this respect; when heated at 38° C.
for fifteen minutes it loses its toxicity, but it then possesses an
immunizing power against the poisons.

The subject possesses great interest, because it was in studying
these immunizing properties that Camus and Gley,[164] and later
on Kossel[165] and Tchistowitch,[166] discovered the first
anticytotoxin,[167] which they obtained by treating the animals with
increasing quantities of the serum of eels. On mixing the antitoxic
serum of these animals _in vitro_ with the red blood-corpuscles of the
species furnishing the serum and of the hemolytic serum of eels, it is
found that the blood-corpuscles kept quite well.

 [164] _Archives internat. de Pharmacodynamie_, III and IV.

 [165] _Berliner Klin. Wochenschr._, 1895, No. 7.

 [166] _Annal. de l'Instit. Pasteur_, XIII, p. 406.

 [167] The name "cytases" or "alexins" has been given to hemolyzing
 diastatic substances which are found in certain serums. It has been
 known for a long time that the serum of the blood of many animals
 destroys the red blood-corpuscles of other and different species. The
 chemical composition of these cytases or alexins is not yet definitely
 known, but the substances rank among the albuminoids; they are
 destroyed by a temperature of 55° to 56° C., and act only in saline
 solutions (Ehrlich and Morgenroth, _Berlin. Klin. Woch._, pp. 6 and
 481). The cytases or alexins, which will be studied in another volume
 of this collection, and which will discuss the active principles
 of the immunizing serums, constitute one of the numerous soluble
 intraleucocytary ferments, and they pass into the serous liquids of the
 organism only as the result of a rupture of or injury to the phagocytes.

As to the blood of the hedgehog, we have already seen that Physalix
and Bertrand have shown that it may be a counter-poison towards
serpent-venom under certain conditions. In its normal condition it is
highly toxic.

=Poisonous Meats.=--It is particularly among the fish that we find
these normally present, and it is a singular fact that, for a given
species, the toxicity frequently depends upon the period of the
year. Thus, at the period of spawning, certain fish may be extremely
poisonous, or, on the contrary, may entirely cease to be so. The
anchovy ballassa from the shores of India occasions death even in
very small quantity; the poisonous meltite of the same seas causes
violent vomiting; the fugu of the Japanese seas possesses an extreme
poisonousness at the spawning period, while, on the contrary, it is
perfectly innocuous at all other periods.

Numerous cases of poisoning have been chronicled every year by the
journals, due to the ingestion of mussels; in the flesh of these
crustaceæ is found a dangerous toxin, _methylotoxin_. The flesh of
oysters is also unwholesome at the spawning period.

The toxic symptoms caused by these animals become apparent in not
less than twenty-four hours after ingestion. The poisoning due to
these fresh meats must not, however, be confounded with that caused by
tainted or spoiled meats.




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