iu which the advance of science has led to the establishment of the method in question. This has been dealt with in a communication made iu the Examination Hall in London and published in the English Medical Journals. The object of the present meeting is to place before you the experimental side of the question and to make you acquainted with the facts already established, and to estimate the grounds of the hope with which we introduce this method into actual practice.
Original {fl-oinmuntqafion;;.
The choleraic
The microbe
infection of animals.
to which we attribute the cause
Cholera is the comma-bacillus of that this malady is attributed to the microbe in is that it is the only question By Moifs. W. M. IIaffkine. one which is constantly found in the intestines Lecture delivered in the Calcutta Medical College on the of cholera patients and never in those of other 2ith March 1893. diseases or of healthy men. This discovery first Since the discovery of the agents of infecmude in Egypt, verified afterwards in Syria and tious diseases and of the power of production of in the epidemic of Spain, iu that of these diseases at will, the manner of introducing India, France, Italy and Massowah, and in the last Formernew methods has profoundly changed. of Russia, Germany and Paris, has epidemics ly the physician could only be guided by conjec- led to the conclusion that the specific poison of when tures and theoretical conclusions, which, cholera is produced solely by this bacillus, and they had led him to an hypothesis possessing that the other microbes found associated with some degree of probability, he was only able it, and which belong to the habitual fauna of Tiiose who have atto test on diseased men. the intestine only serve to favour or hinder its well the great diffiknow innovations tempted entrance into the intestinal canal. culties lying in the path of success. It may be The comma bacillus, propagated in the said in a general way that never is a purely theoretical conclusion fully confirmed by ex- laboratory by a series of cultures, shows itself to he variable in form, character of culture, perience. The conditions which determine the and virulence. The appearance of the culture and diverse. multifarious so phenomena are Most of the methods employed in medicine have in gelatine, generally the most characteristic, debeen discovered accidentally notwithstanding pends upon the fact that this microbe, like many others, secretes a special ferment which dissolves all the attempts at a priori reasoning. Only this substance. The secretion of this ferment two branches of knowledge, astronomy and not constant, and often a microbe is, however, have optics, (sciences purely mathematical,) which during many years preserves this fagiven us two or three great discoveries foreseen culty of liquifying gelatine intact, loses it at by speculative mind?. a On moment iu a relatively short time. of Siuce the discovery of the means reproduc- thegiven other haud, the virulence of the microbe for of disease in the animals things ing aspect medicine has changed. With the discovery of depends on the secretion of a special poison?a power of producing which is also pathological microbes, almost all the diseases toxine, the lost with known infectious agents can be communi- generally during the course of its cultivation in the cated to animals. laboratory. It is necessary to insist A.t present our methods are no the fact that the toxines have nothing iu upon confined into to abstract ideas, but enter longer common with the ferment liquefying the gelathe domain of objective facts. We can experitine in these substances have certain cases ; on animals ment by conveying to them the disbeen and studied apart. meanwhile artificially for separated ease, seeking, many years, efficaciThe microbe of cholera passes a great part ous measures of treatment, and only approaching our of its life outside the organism of man; iu the when in man experiments the laboratory have Vaccination against cholera is soil, in the water, in the air. With the vicissibeen successful. at this stage. By experimental researches a pro- tudes and varying conditions of its saprophyphylactic procedure has been established, the tic life, this microbe changes in its properties, efficacy of which can be demonstrated at any both in those which act on the culture medium moment. Necessarily the practical physician aud in those which cause the disease in man. In has to assure himself of its value, and before Nature as well as in the laboratory the microbe putting it in practice, to demand the most abso- modifies or loses its power of secreting the lute proof of its efficacy. I cannot enter into ferment which liquefies gelatine, and also its Still every the history of the subjeot, or into the maimer power of secreting the toxine.
VACCINATION AGAINST ASIATIC CHOLERA.
ot Asiatic
Koch. The
reason
-
13
98
INDIAN MEDICAL GAZETTE.
time we search for the microbe, not in its natural habitat, but in the intestine of a cholera patient, we must obviously expect to finil a specimen which,varying as much as possible in all its other properties, will have preserved intact its power of giving the disease. While dealing with our subject it is useful to put aside for the time the other properties of this microbe which concern the biologist and the morphologist, and to give the name of Asiatic choleraic microbe to specimens which have preserved the pathological property of producing this malady. It is important to know that, in this relationship comma-bacilli have a nature essentially identical. In effect we shall see later that animals protected by choleraic vaccine resist equally the different varieties of cholera microbes obtained under the most variable con-
ditions. It appears that all animals used in the laboraare sensitive to the choleraic poison. Perthe of virulence haps by accentuating sufficiently the comma-bacillus no animal will be found proof against it. However, in order to experiment, with cholera as an infectious disease, it is not sufficient to be able to poison the animal by the poison itself; it is necessary to cause the microbe to live and multiply in the interior of the animal organism, so that it may be in a position to struggle with the cellules of the latter, to be able to pass it from one animal to another, and to produce the disease in an infinite series of subjects of the same species. As we find it in Nature or cultivated iu the organism of man, the microbe of cholera cannot pass into the bodies of inferior auimals. In order to accomplish this it is necessary to modify gradually its physiological habits. The process by which this end is arrived at is to inject a strong dose of these microbes into the peritoneal cavity of guinea-pigs. The animal dies poisoned iu the space of 15 to 20 hours, after which there is found in this cavity a quantity of exudation containing comma-bacilli. This liquid is removed with care, and exposed to oxidation in the air for several hour?, and afterwards injected into the peritoneum of another animal which is selected in accordance with its size in inverse proportion to the quantity of liquid removed from the body of the first animal. For a large quantity of fluid a small animal is chosen, and for a small quantity a larger one. By transferring the effusion from a second to a third animal, from a third to a fourth, and so on, the series can be continued indefinitely. In proportion as we advance, the infectious properties of the microbe become more and more exalted. The dose of the microbes
tory
to kill the animal will diminish, death will occur more speedily, and the leucocyte The reaction will be gradually suppressed. microbe will acquire the property of killing the most resisting kinds of animal*, while those
necessary
which have attained
[April immunity by
the
1893.
specia
process of inoculation against this microbe will preserve their immunity during long periods. We thus obtain a complete picture of a most characteristic microbian disease. It is important to observe that in proceeding in this manner we do not produce a new toxinesecreting function in the microbe, but only increase that which existed from the beginning, and which it is easy to show existed before any passage through the animal. Besides this, we communicate to it the property of living and multiplying in the organism of that animal. These are the two modifications which the procedure of " passage" imports into the physiology of the microbe. In the natural disease in man the microbe is confined to the small intestine, and does not penetrate into the peritoneal cavity. It is iu the intestine that it multiplies ami secretes its toxine, and it is from there that this toxine passes into the rest of the organism and poisons the system. With certain precautions it is easy to' reproduce this same form of intoxication in animals ; only it is less severe than when the microbe and its toxine act ou the peritoneal cavity. The reason is probably simple. In the intestine the microbe and the toxine of cholera are mixed with an infinite variety of microbes and other ferments which destroy and neutralise their action. Here are two dead guineapigs, and one rabbit killed by the intraperiAfter the toneal infection, and a sick animal. meeting we will make an autopsy of this animal, and we shall find in the peritoneal cavity the exudation in question full of comma-bacilli, the small intestine will be inflamed and its contents liquefied. The spleen and liver will be found iu a condition reciprocally inversed. In the case of a virulent microbe and a feebly resisting animal, the liver has a normal aspect and the spleen is enlarged and black. In the case of a feeble microbe, and a highly resisting animal the spleen is normal, but the liver is covered with a fibrinous deposit formed of white blood corpuscles. Occasionally microbes penetrate into the thoracic cavity and into the intestines, the internal muscles of the sternum are strongly inflamed, and the bladder is empty. When the microbe has arrived at its maximum power the animal dies 8 hours after infection. The fatal dose is small, and with the quantity of an ordinary agar-agar culture one can kill 32 to 64 individuals. The inoculation of this microbe deep into the muscle produces an enor mous swelling and finally kills the animal. The inoculation of the peritoneal cavity of rabbits and pigeons is mortal for the latter. The Choleraic Vaccination.
Tim cholera vaccination method has been established by following the way traced in this
April
1893.]
HAFFKINE ON VACCINATION AGAINST ASIATIC CHOLERA.
Since we possess direction by vaccination against smallpox and cholera, brought by by that against rabies. It consists in the hypo-pigs to a very high dermic inoculation of a virus of a definite natureinoculate it
already the
99
Original {fl-oinmuntqafion;;.
The choleraic
The microbe
infection of animals.
to which we attribute the cause
Cholera is the comma-bacillus of that this malady is attributed to the microbe in is that it is the only question By Moifs. W. M. IIaffkine. one which is constantly found in the intestines Lecture delivered in the Calcutta Medical College on the of cholera patients and never in those of other 2ith March 1893. diseases or of healthy men. This discovery first Since the discovery of the agents of infecmude in Egypt, verified afterwards in Syria and tious diseases and of the power of production of in the epidemic of Spain, iu that of these diseases at will, the manner of introducing India, France, Italy and Massowah, and in the last Formernew methods has profoundly changed. of Russia, Germany and Paris, has epidemics ly the physician could only be guided by conjec- led to the conclusion that the specific poison of when tures and theoretical conclusions, which, cholera is produced solely by this bacillus, and they had led him to an hypothesis possessing that the other microbes found associated with some degree of probability, he was only able it, and which belong to the habitual fauna of Tiiose who have atto test on diseased men. the intestine only serve to favour or hinder its well the great diffiknow innovations tempted entrance into the intestinal canal. culties lying in the path of success. It may be The comma bacillus, propagated in the said in a general way that never is a purely theoretical conclusion fully confirmed by ex- laboratory by a series of cultures, shows itself to he variable in form, character of culture, perience. The conditions which determine the and virulence. The appearance of the culture and diverse. multifarious so phenomena are Most of the methods employed in medicine have in gelatine, generally the most characteristic, debeen discovered accidentally notwithstanding pends upon the fact that this microbe, like many others, secretes a special ferment which dissolves all the attempts at a priori reasoning. Only this substance. The secretion of this ferment two branches of knowledge, astronomy and not constant, and often a microbe is, however, have optics, (sciences purely mathematical,) which during many years preserves this fagiven us two or three great discoveries foreseen culty of liquifying gelatine intact, loses it at by speculative mind?. a On moment iu a relatively short time. of Siuce the discovery of the means reproduc- thegiven other haud, the virulence of the microbe for of disease in the animals things ing aspect medicine has changed. With the discovery of depends on the secretion of a special poison?a power of producing which is also pathological microbes, almost all the diseases toxine, the lost with known infectious agents can be communi- generally during the course of its cultivation in the cated to animals. laboratory. It is necessary to insist A.t present our methods are no the fact that the toxines have nothing iu upon confined into to abstract ideas, but enter longer common with the ferment liquefying the gelathe domain of objective facts. We can experitine in these substances have certain cases ; on animals ment by conveying to them the disbeen and studied apart. meanwhile artificially for separated ease, seeking, many years, efficaciThe microbe of cholera passes a great part ous measures of treatment, and only approaching our of its life outside the organism of man; iu the when in man experiments the laboratory have Vaccination against cholera is soil, in the water, in the air. With the vicissibeen successful. at this stage. By experimental researches a pro- tudes and varying conditions of its saprophyphylactic procedure has been established, the tic life, this microbe changes in its properties, efficacy of which can be demonstrated at any both in those which act on the culture medium moment. Necessarily the practical physician aud in those which cause the disease in man. In has to assure himself of its value, and before Nature as well as in the laboratory the microbe putting it in practice, to demand the most abso- modifies or loses its power of secreting the lute proof of its efficacy. I cannot enter into ferment which liquefies gelatine, and also its Still every the history of the subjeot, or into the maimer power of secreting the toxine.
VACCINATION AGAINST ASIATIC CHOLERA.
ot Asiatic
Koch. The
reason
-
13
98
INDIAN MEDICAL GAZETTE.
time we search for the microbe, not in its natural habitat, but in the intestine of a cholera patient, we must obviously expect to finil a specimen which,varying as much as possible in all its other properties, will have preserved intact its power of giving the disease. While dealing with our subject it is useful to put aside for the time the other properties of this microbe which concern the biologist and the morphologist, and to give the name of Asiatic choleraic microbe to specimens which have preserved the pathological property of producing this malady. It is important to know that, in this relationship comma-bacilli have a nature essentially identical. In effect we shall see later that animals protected by choleraic vaccine resist equally the different varieties of cholera microbes obtained under the most variable con-
ditions. It appears that all animals used in the laboraare sensitive to the choleraic poison. Perthe of virulence haps by accentuating sufficiently the comma-bacillus no animal will be found proof against it. However, in order to experiment, with cholera as an infectious disease, it is not sufficient to be able to poison the animal by the poison itself; it is necessary to cause the microbe to live and multiply in the interior of the animal organism, so that it may be in a position to struggle with the cellules of the latter, to be able to pass it from one animal to another, and to produce the disease in an infinite series of subjects of the same species. As we find it in Nature or cultivated iu the organism of man, the microbe of cholera cannot pass into the bodies of inferior auimals. In order to accomplish this it is necessary to modify gradually its physiological habits. The process by which this end is arrived at is to inject a strong dose of these microbes into the peritoneal cavity of guinea-pigs. The animal dies poisoned iu the space of 15 to 20 hours, after which there is found in this cavity a quantity of exudation containing comma-bacilli. This liquid is removed with care, and exposed to oxidation in the air for several hour?, and afterwards injected into the peritoneum of another animal which is selected in accordance with its size in inverse proportion to the quantity of liquid removed from the body of the first animal. For a large quantity of fluid a small animal is chosen, and for a small quantity a larger one. By transferring the effusion from a second to a third animal, from a third to a fourth, and so on, the series can be continued indefinitely. In proportion as we advance, the infectious properties of the microbe become more and more exalted. The dose of the microbes
tory
to kill the animal will diminish, death will occur more speedily, and the leucocyte The reaction will be gradually suppressed. microbe will acquire the property of killing the most resisting kinds of animal*, while those
necessary
which have attained
[April immunity by
the
1893.
specia
process of inoculation against this microbe will preserve their immunity during long periods. We thus obtain a complete picture of a most characteristic microbian disease. It is important to observe that in proceeding in this manner we do not produce a new toxinesecreting function in the microbe, but only increase that which existed from the beginning, and which it is easy to show existed before any passage through the animal. Besides this, we communicate to it the property of living and multiplying in the organism of that animal. These are the two modifications which the procedure of " passage" imports into the physiology of the microbe. In the natural disease in man the microbe is confined to the small intestine, and does not penetrate into the peritoneal cavity. It is iu the intestine that it multiplies ami secretes its toxine, and it is from there that this toxine passes into the rest of the organism and poisons the system. With certain precautions it is easy to' reproduce this same form of intoxication in animals ; only it is less severe than when the microbe and its toxine act ou the peritoneal cavity. The reason is probably simple. In the intestine the microbe and the toxine of cholera are mixed with an infinite variety of microbes and other ferments which destroy and neutralise their action. Here are two dead guineapigs, and one rabbit killed by the intraperiAfter the toneal infection, and a sick animal. meeting we will make an autopsy of this animal, and we shall find in the peritoneal cavity the exudation in question full of comma-bacilli, the small intestine will be inflamed and its contents liquefied. The spleen and liver will be found iu a condition reciprocally inversed. In the case of a virulent microbe and a feebly resisting animal, the liver has a normal aspect and the spleen is enlarged and black. In the case of a feeble microbe, and a highly resisting animal the spleen is normal, but the liver is covered with a fibrinous deposit formed of white blood corpuscles. Occasionally microbes penetrate into the thoracic cavity and into the intestines, the internal muscles of the sternum are strongly inflamed, and the bladder is empty. When the microbe has arrived at its maximum power the animal dies 8 hours after infection. The fatal dose is small, and with the quantity of an ordinary agar-agar culture one can kill 32 to 64 individuals. The inoculation of this microbe deep into the muscle produces an enor mous swelling and finally kills the animal. The inoculation of the peritoneal cavity of rabbits and pigeons is mortal for the latter. The Choleraic Vaccination.
Tim cholera vaccination method has been established by following the way traced in this
April
1893.]
HAFFKINE ON VACCINATION AGAINST ASIATIC CHOLERA.
Since we possess direction by vaccination against smallpox and cholera, brought by by that against rabies. It consists in the hypo-pigs to a very high dermic inoculation of a virus of a definite natureinoculate it
already the
99
