By Weakened Virus. This has been secured by heating the defibrinated blood to 55° C. for ten minutes (Toussaint); Pasteur, Chamberland and Roux accomplished the same end by making anthrax cultures at 42° to 43° C. in presence of air; Chauveau by subjecting the virulent culture for eight days to oxygen under a pressure of 8 atmospheres at a temperature of 38° C.; Chamberland, Roux and others have cultivated the bacillus in weak antiseptic bouillons as phenic acid (1:600 or 1200), bichromate of potash (1:2000 or 5000), sulphuric acid (2:100).
Other methods have been followed, as growing the bacillus in the blood or serum of immune animals (dog, chicken, pigeon, white rat, frog).
Of these different methods that of Pasteur has been most extensively adopted. The temperature of culture (42° C.) prevents the formation of spores and the duration of exposure to air gradually lessens the virulence until in 12 or 13 days it is not fatal to the Guinea pig and after 31 days it fails to kill the young mouse. Thus preparations of varying grades of virulence, and adapted to the varying susceptibility of different animals, are secured. The protective inoculation is made by preference in spring, when there is less chance of complication by a coincident accidental infection, it is to be avoided if possible in animals at hard work, in advanced gestation, in full milk, in extreme youth, or in ill health. To secure the best results it should be repeated with a stronger preparation 12 to 15 days after the first injection. The acquired immunity lasts a year and over, and it is probably perpetuated by new and non-fatal doses taken in casually, on the anthrax pastures. Hundreds of thousands of live stock in all parts of the world have been treated in this way with the result of reducing a mortality of two, five or ten per cent. to insignificant proportions. It can only be safely adopted on anthrax lands, as elsewhere it may lead to the stocking of new areas with a malignant germ which in young and susceptible animals reacquires its original virulence.
It can never be safely ignored that we are dealing with the living seed of a most deadly infection. Though robbed of a large part of its virulence by artificial culture at 107:5° F., yet many accidental conditions, contribute to a relapse to its original potency, and when it has once killed a victim, the reacquired virulence is usually persistent. If the virus employed for protective purposes in cattle and sheep, is inoculated on Guinea pigs of 1 to 30 days old, from these on those of several months, and from these last on sheep, the virulence is constantly and persistently enhanced. The same is true of the microbe which is inoculated on a succession of pullets of steadily encreasing ages (Roux and Chamberland), or on a succession of pigeons (Metchnikoff). The germs reinforced in potency in any such way are liable to be the starting points for dangerous infections in animals and permanent contamination of soils and waters. Fortunately an occurrence of this kind is rare, yet with a wide application of the Pasteurian inoculation the opportunities also are great, and with the free sale and distribution of the mitigated virus (anthrax “vaccine”), the evil may grow indefinitely. The method departs from the ideal one, aimed at a final extinction of the disease, and accepts in place a mere temporary protection of the herd or flock, and though in this affection eradication cannot always be secured, every effort should be made to gain it and above all to prevent an encrease of the area of infection.
Technique. The weakened virus (1st “vaccin”) is sold in tubes holding enough for 100, 200, or 300 sheep. Of this ⅛ cc. is injected subcutem on the inner side of the thigh of the mature sheep, and 12 or 15 days later a similar dose of the stronger preparation (“2d vaccin”). For the ox or horse double the amount (¼th cc.) is used, being injected behind the shoulder, and on the side of the neck in the respective animals. The dose is graduated in the different subjects according to the size and age, yet a considerable latitude is permissible. The syringe must be disinfected before and after inoculations by a 5 per cent. solution of carbolic acid, or by boiling, and the nozzle should be dipped in strong carbolic acid immediately before and after each insertion. This will greatly obviate infection of the liquid used, and of the wound by any virulent germs lodged on the surface of the skin. The liquid to be injected should be used as soon as possible after preparation, and if kept should be in a dark cold place, and if the tube is once opened the whole of its contents should be used the same day,—never kept over. The second, stronger preparation should never be used until the system has been prepared for it by the use of the first.
By the Soluble Toxins in Sterile Solution. In 1884, in an outbreak of anthrax in Skaneateles, N. Y., I drew blood from an anthrax cow, subjected it to 212° F. for 30 minutes, dissolved out the soluble toxins in boiled water, and injected the product subcutem, in the dose of 2 to 4cc. according to size, into every apparently healthy member of the herd, excepting one, which was left as a check. The check animal died of anthrax while all of the others escaped.
Since that time I have personally used it in every herd where opportunity offered, and with equally good results. In an outbreak near Elmira, Dr. Moore adopted it in a large dairy herd, and the disease was suddenly arrested.
In several experimental cases (one cow and 2 Guinea pigs) at the N. Y. S. V. College, the outcome was not so satisfactory and in a herd in Oneida Co., N. Y., it is said to have failed to check the disease.
Notwithstanding these untoward results in other hands, I am still confident that we have in this a measure of no small value, and worthy of application in suitable cases. A certain percentage of failures in immunization are to be looked for. Even cowpox vaccination is not always protective against itself; I knew one man who was successfully vaccinated every three years in a comparatively long series. Many habitually self-limiting diseases relapse in particular individuals. Even in the case of anthrax excess of glucose in the system, and the lack of some unknown influence of the spleen are respectively destructive of immunity. Even after the Pasteurian inoculation a certain number of inoculated animals are lost, it may be between the first and second injection, or it may be “two or three months” after the latter (Galtier). We must also bear in mind that in an infected herd or flock, there are always a certain number already infected at the time of the protective inoculation, and as the protective conditions are slowly established, through the action of the leucocytes, it is unreasonable to expect that serious illness and death can be obviated in such animals.
In inoculation with the Pasteur lymph, the bacillus is held not to enter the blood, a position supported by the researches of Bitter, Perroncito, Wissokovicz, Lubarsch, Metchnikoff, Chamberland and Roux, so that the resulting immunization must come from the toxins. Add to this that Chauveau (1885), conferred immunity on a sheep by injecting intravenously, anthrax blood, defibrinated and sterilized by heat: Arloing obtained immunity in the sheep by injecting, subcutem, the clear supernatant liquid from old bouillon cultures of anthrax, from which all bacilli had been precipitated. Roux and Chamberland obtained the same result by using the pulp of an anthrax spleen treated with essential oil of mustard, so as to destroy the life of the bacillus, and then evaporated in vacuo to remove the essence. Smaller doses proved effective, than when the splenic pulp had been filtered or sterilized by heating to 58° C.