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Antitoxic serum in the treatment of anthrax
ABSTRACTS AND REPORTS.
16 7
Third Series.-Using the same precautions as in the first series, a monkey was inoculated with the contents of pustules from a case of small-pox in the human subject; seven days afterwards excavated papules surrounded by an areola had formed, and there were also some papules on the lips and extremities. ThlS generalisation was produced only once in seven such experiments, but in everyone of the monkeys inoculated some pustules formed. The monkey may thus be infected with small-pox of the human subject, and this variolation as a rule gives rise to only a local eruption. Fourtlt ~eries.-A monkey was inoculated with the contents of one of the pustules of a monkey of the first series. This second passage gave good pustules after seven days, and the same result up to the seventh passage. Rjtlz Serzts.- The monkeys of the fourth series were inoculated \\,lth fresh retrovaccin after the crusts had fallen. Result negative. The retrovaccin cultivated by successive passages in the monkey retained its infective power and its immunising power. Sixth Series.-The seven monkeys of the second series and the six monkeys of the fifth series were inoculated with the contents of pustules from a case of smallpox. Result negative. Retrovaccin from the calf as well as retrovaccin cultivated by seven passages on the monkey thus preserves the monkey against small-pox. Sez'oztll Series.- The small-pox of the first monkey of the third series was inoculated on a second monkey, and one obtained only pustules localised at the point of inoculation. The third, fourth, fifth, sixth, and seventh passages gave similar pustules, always in seven days. This series of experiments was repeated thrice with the same results. Eigltth Series.-With the sixth passage of the seventh series a calf was inoculated; after five days this calf showed pustules which in all respects resembled those ordinarily obtained with calf-vaccin. This experiment has be~n repeated with the same result with the seventh passage of the seventh senes. Ninth Serz'es.-The second calf of the eighth series was inoculated seven days after the first inoculation with fresh retrovaccin. No result. From these experiments one may draw the important conclusion that the retrovaccin protects the organism of the monkey against the virus of small-pox; this fact is in favour of the identity of the retrovaccin with the ordinary vaccin, which produces the same effect. The author remarks that at the present time we know nothing about the origin of cow-pox or of the relationship between that,'and small-pox, but he inclines to the view that true cow-pox, mitigated small-pox, and retrovaccin are identical; for each of these three viruses: (I) Provokes in the human subject after seven days similar pustules, with more or less fever; (2) protects against small-pox; (3) changes the duration of development when one inoculates it to various species of animals; and (4) degenerates after a greater or smaller number of passages on animals of the same species. The author admits that the most convincing proof of the identity of smallpox and cow-pox would have been furnished by using matter from the mitigated small-pox of the monkey for vaccinating human beings; that, however. was an experiment that he did not feel justified in performing.-.:lmzalt's de l'Institut Pasteur.
ANTITOXIC SERUM IN THE TREATMENT OF ANTHRAX. IN the month of January 1887, in the first number of the Anllales de l'Illstitut Pasteur, Metchnikoff stated that cultures of the anthrax bacillus made in the blood of vaccinated sheep had lost their virulence to such a
168
ABSTRACTS AND REPORTS.
degree that a cubic centimetre of the culture was incapable of causing the death of a rabbit. This fact raised the question whether the bacilli had been attenuated by contact with the serum, or whether the serum rendered the animal insusceptible to anthrax. The discovery of the preventive property of the serum of immunised animals has again raised this problem, and Marchoux has devised and carried out a series of experiments to show whether the serum of animals that have been immunised against living and very virulent anthrax cultures is preventive and curative. His experiments were carried out partly on rabbits and partly on sheep. When experiments have to be carried out on a large scale it is much better to immunise animals of the latter species. as they furnish much more blood. The sheep were first vaccinated according to the method of Pasteur, and they then had injected under the skin progressively increasing doses of virulent anthrax cultures; the quantity was doubled every eight days, and ultimately a dose of 200 cc. or even 300 cc . .of a very active culture were injected at one time. "With such enormous doses the animals exhibited a very marked reaction, manife'ited by diminution in weight and an elevation of temperature which sometimes lasted for eight days. "When the culture was injected into the veins the effect was scarcely more severe than after subcutaneous inoculation, provided only a small quantity of virus was used, but the intravenous inoculations were much more dangerous when large doses were employed. A sheep vaccinated in the ordinary way is quite refractory to inoculation with anthrax, even when large doses are employed, but the serum which such an animal furnishes has scarcely any preventive properties and no curative power at all. In order to develop the latter it is necessary to bring about a very high degree of immunity, so that the animals will bear the enormous doses which it is necessary to inject from time to time. As regards the proper moment at which to bleed an animal in order to obtain the most efficacious serum, it was ascertained by experiment that the curative power of the serum is most marked at from fifteen days to three weeks after the injection of virulent cultures. The first sheep was bled on the 18th January, and its serum when tested was found to have no preventive power. Two rabbits, one of which received 12 cc. and the other 20 cc. died after the same lapse of time as the control animal. The sheep was bled again on the 28th May 1895; it had been under treatment since 18th January, when it received about 200 cc. of culture, and the last inoculation had taken place on the 18th May with 100 cc. On the 29th May three rabbits were inoculated, the first with 2 cc. of serum, the second with + cc., and the third with 8 cc.; on the following day each of them received at the same time as a control animal one-third of a cubic centimetre of virulent anthrax culture. The control animal died in twenty-four hours, and rabbit No. I in eighteen days, while rabbits Nos. 2 and 3 survived. This experiment was repeated on several occasions, and always with most identical results. With a dose of 3 cc. the life of the animal was always saved and several times this result was obtained with 2 cc.; on the other hand, a dose of ICC. never sufficed to protect. The second sheep, after it had received 1050 cc. of virulent anthrax culture, furnished a serum which was only a little stronger than that of the first sheep. The dose which certainly protected had been reduced to 2 cc. ; and sometimes I cc. was preventive. "\Vhen this second sheep had received a total of 1400 cc. of virulent culture the preventive power of its serum had risen to such a degree that icc. was preventive. This is the most active serum that Marchoux has been able to obtain, but he thinks that it might be obtained of a higher degree of activity by using in place of sheep larger and more sensitive animals, such as the ass or the horse.
ABSTRACTS AND REPORTS.
The experiments just described ~how that the serum of the sheep is possessed of distinctly preventive powers against anthrax; but none of the animals treated by this serum had been v'1.Ccinated; all succumbed to a subsequent inoculation with virulent anthrax. In these experiments the inoculation was made under the skin of the flank, and the serum was injected on the one side and the anthrax on the other. It was found that the inoculation was more or less grave according to the point of the body where it was made, and in order to obtain protection a larger or smaller dose of serum was necessary according to the seat of the inoculation. Thus, in order to preserve a rabbIt inoculated under the skin of the ear required twice as much serum as when the inoculation was made under the skin of the back. A certain number of experiments were made on guinea-pigs with the serum, but in no instance was a successful result obtained. Ten cc. of serum injected into the peritoneum delayed death twelve hours after that of the control animals, and this was the best result obtained in the case of guinea-pigs. Experiments were also made to test the curative power of the serum. It was ascertained that the injection of a sufficient dose of serum immediately after inoculation with the virus sufficed to prevent development of any symptoms of the disease, just as if the bacilli introduced under the skin had not been able to develop. These animals were not vaccinated, for when subsequently inoculated they died from anthrax at the same time as the control subjects. The result is not the same when an interval is allowed to elapse between inoculation with the virus and the injection of the serum; in the case where the serum was injected into rabbits in from seven to twentyfour hours after infection, the animals showed premonitory symptoms of the disease, and when the animals were subsequently tested twelve days afterwards it was found that they had acquired a very solid resistance to anthrax. When there is already considerable cedema at the seat of moculation before the serum is injected it is very difficult to save the animal's life, even when considerable doses are employed; the best result obtained in such a case was the survival of a day or so after the control subjects. It was noted, however, in the case of some of the animals that twelve or eighteen hours after the adm1l1istration of the serum there was a distinct diminution in the extent of the local cedema or even a complete disappearance of it; nevertheless, these animals died with numerous bacilli in the blood, but Marchoux thinks that the result holds out hope that by obtaining a more active serum it may be possible to save the animal's life even at this period of the disease. The immunity acquired by vaccination does not at all resemble that which is brought about by antitoxic serum; the one is profound and lasting, the other superficial and transient. Vaccinated animals can support enormous doses of virulent cultures; thus, a rabbit vaccinated a year previously, and which had not for eight months received any inoculations, resisted inoculation with a dose of 5 cc. of culture, and this, although the pre\'ious inoculations had never e?,ceeded I cc. of virulent culture at a tittle. These facts throw into strong relief the difference between the immunity, difficult to acquire but very durable, which is conferred by vaccination with anthrax VIruS, and the almost immediately acquired but temporary immunity which follows the injection of anticarbon serum.-A71llales de l'Illstitut Pastellr.
16 7
Third Series.-Using the same precautions as in the first series, a monkey was inoculated with the contents of pustules from a case of small-pox in the human subject; seven days afterwards excavated papules surrounded by an areola had formed, and there were also some papules on the lips and extremities. ThlS generalisation was produced only once in seven such experiments, but in everyone of the monkeys inoculated some pustules formed. The monkey may thus be infected with small-pox of the human subject, and this variolation as a rule gives rise to only a local eruption. Fourtlt ~eries.-A monkey was inoculated with the contents of one of the pustules of a monkey of the first series. This second passage gave good pustules after seven days, and the same result up to the seventh passage. Rjtlz Serzts.- The monkeys of the fourth series were inoculated \\,lth fresh retrovaccin after the crusts had fallen. Result negative. The retrovaccin cultivated by successive passages in the monkey retained its infective power and its immunising power. Sixth Series.-The seven monkeys of the second series and the six monkeys of the fifth series were inoculated with the contents of pustules from a case of smallpox. Result negative. Retrovaccin from the calf as well as retrovaccin cultivated by seven passages on the monkey thus preserves the monkey against small-pox. Sez'oztll Series.- The small-pox of the first monkey of the third series was inoculated on a second monkey, and one obtained only pustules localised at the point of inoculation. The third, fourth, fifth, sixth, and seventh passages gave similar pustules, always in seven days. This series of experiments was repeated thrice with the same results. Eigltth Series.-With the sixth passage of the seventh series a calf was inoculated; after five days this calf showed pustules which in all respects resembled those ordinarily obtained with calf-vaccin. This experiment has be~n repeated with the same result with the seventh passage of the seventh senes. Ninth Serz'es.-The second calf of the eighth series was inoculated seven days after the first inoculation with fresh retrovaccin. No result. From these experiments one may draw the important conclusion that the retrovaccin protects the organism of the monkey against the virus of small-pox; this fact is in favour of the identity of the retrovaccin with the ordinary vaccin, which produces the same effect. The author remarks that at the present time we know nothing about the origin of cow-pox or of the relationship between that,'and small-pox, but he inclines to the view that true cow-pox, mitigated small-pox, and retrovaccin are identical; for each of these three viruses: (I) Provokes in the human subject after seven days similar pustules, with more or less fever; (2) protects against small-pox; (3) changes the duration of development when one inoculates it to various species of animals; and (4) degenerates after a greater or smaller number of passages on animals of the same species. The author admits that the most convincing proof of the identity of smallpox and cow-pox would have been furnished by using matter from the mitigated small-pox of the monkey for vaccinating human beings; that, however. was an experiment that he did not feel justified in performing.-.:lmzalt's de l'Institut Pasteur.
ANTITOXIC SERUM IN THE TREATMENT OF ANTHRAX. IN the month of January 1887, in the first number of the Anllales de l'Illstitut Pasteur, Metchnikoff stated that cultures of the anthrax bacillus made in the blood of vaccinated sheep had lost their virulence to such a
168
ABSTRACTS AND REPORTS.
degree that a cubic centimetre of the culture was incapable of causing the death of a rabbit. This fact raised the question whether the bacilli had been attenuated by contact with the serum, or whether the serum rendered the animal insusceptible to anthrax. The discovery of the preventive property of the serum of immunised animals has again raised this problem, and Marchoux has devised and carried out a series of experiments to show whether the serum of animals that have been immunised against living and very virulent anthrax cultures is preventive and curative. His experiments were carried out partly on rabbits and partly on sheep. When experiments have to be carried out on a large scale it is much better to immunise animals of the latter species. as they furnish much more blood. The sheep were first vaccinated according to the method of Pasteur, and they then had injected under the skin progressively increasing doses of virulent anthrax cultures; the quantity was doubled every eight days, and ultimately a dose of 200 cc. or even 300 cc . .of a very active culture were injected at one time. "With such enormous doses the animals exhibited a very marked reaction, manife'ited by diminution in weight and an elevation of temperature which sometimes lasted for eight days. "When the culture was injected into the veins the effect was scarcely more severe than after subcutaneous inoculation, provided only a small quantity of virus was used, but the intravenous inoculations were much more dangerous when large doses were employed. A sheep vaccinated in the ordinary way is quite refractory to inoculation with anthrax, even when large doses are employed, but the serum which such an animal furnishes has scarcely any preventive properties and no curative power at all. In order to develop the latter it is necessary to bring about a very high degree of immunity, so that the animals will bear the enormous doses which it is necessary to inject from time to time. As regards the proper moment at which to bleed an animal in order to obtain the most efficacious serum, it was ascertained by experiment that the curative power of the serum is most marked at from fifteen days to three weeks after the injection of virulent cultures. The first sheep was bled on the 18th January, and its serum when tested was found to have no preventive power. Two rabbits, one of which received 12 cc. and the other 20 cc. died after the same lapse of time as the control animal. The sheep was bled again on the 28th May 1895; it had been under treatment since 18th January, when it received about 200 cc. of culture, and the last inoculation had taken place on the 18th May with 100 cc. On the 29th May three rabbits were inoculated, the first with 2 cc. of serum, the second with + cc., and the third with 8 cc.; on the following day each of them received at the same time as a control animal one-third of a cubic centimetre of virulent anthrax culture. The control animal died in twenty-four hours, and rabbit No. I in eighteen days, while rabbits Nos. 2 and 3 survived. This experiment was repeated on several occasions, and always with most identical results. With a dose of 3 cc. the life of the animal was always saved and several times this result was obtained with 2 cc.; on the other hand, a dose of ICC. never sufficed to protect. The second sheep, after it had received 1050 cc. of virulent anthrax culture, furnished a serum which was only a little stronger than that of the first sheep. The dose which certainly protected had been reduced to 2 cc. ; and sometimes I cc. was preventive. "\Vhen this second sheep had received a total of 1400 cc. of virulent culture the preventive power of its serum had risen to such a degree that icc. was preventive. This is the most active serum that Marchoux has been able to obtain, but he thinks that it might be obtained of a higher degree of activity by using in place of sheep larger and more sensitive animals, such as the ass or the horse.
ABSTRACTS AND REPORTS.
The experiments just described ~how that the serum of the sheep is possessed of distinctly preventive powers against anthrax; but none of the animals treated by this serum had been v'1.Ccinated; all succumbed to a subsequent inoculation with virulent anthrax. In these experiments the inoculation was made under the skin of the flank, and the serum was injected on the one side and the anthrax on the other. It was found that the inoculation was more or less grave according to the point of the body where it was made, and in order to obtain protection a larger or smaller dose of serum was necessary according to the seat of the inoculation. Thus, in order to preserve a rabbIt inoculated under the skin of the ear required twice as much serum as when the inoculation was made under the skin of the back. A certain number of experiments were made on guinea-pigs with the serum, but in no instance was a successful result obtained. Ten cc. of serum injected into the peritoneum delayed death twelve hours after that of the control animals, and this was the best result obtained in the case of guinea-pigs. Experiments were also made to test the curative power of the serum. It was ascertained that the injection of a sufficient dose of serum immediately after inoculation with the virus sufficed to prevent development of any symptoms of the disease, just as if the bacilli introduced under the skin had not been able to develop. These animals were not vaccinated, for when subsequently inoculated they died from anthrax at the same time as the control subjects. The result is not the same when an interval is allowed to elapse between inoculation with the virus and the injection of the serum; in the case where the serum was injected into rabbits in from seven to twentyfour hours after infection, the animals showed premonitory symptoms of the disease, and when the animals were subsequently tested twelve days afterwards it was found that they had acquired a very solid resistance to anthrax. When there is already considerable cedema at the seat of moculation before the serum is injected it is very difficult to save the animal's life, even when considerable doses are employed; the best result obtained in such a case was the survival of a day or so after the control subjects. It was noted, however, in the case of some of the animals that twelve or eighteen hours after the adm1l1istration of the serum there was a distinct diminution in the extent of the local cedema or even a complete disappearance of it; nevertheless, these animals died with numerous bacilli in the blood, but Marchoux thinks that the result holds out hope that by obtaining a more active serum it may be possible to save the animal's life even at this period of the disease. The immunity acquired by vaccination does not at all resemble that which is brought about by antitoxic serum; the one is profound and lasting, the other superficial and transient. Vaccinated animals can support enormous doses of virulent cultures; thus, a rabbit vaccinated a year previously, and which had not for eight months received any inoculations, resisted inoculation with a dose of 5 cc. of culture, and this, although the pre\'ious inoculations had never e?,ceeded I cc. of virulent culture at a tittle. These facts throw into strong relief the difference between the immunity, difficult to acquire but very durable, which is conferred by vaccination with anthrax VIruS, and the almost immediately acquired but temporary immunity which follows the injection of anticarbon serum.-A71llales de l'Illstitut Pastellr.