TETANUS. 61 



virulence on being boiled with alcohol rendered feebly alkaline. 

 The virulence of a culture and the amount of this substance con- 

 tained therein are in direct proportion to each other. Small doses 

 (one milligram to one hundred grams of body weight) in feebly 

 alkaline solution, introduced into the stomachs of guinea-pigs cause, 

 as a rule, within from four to six hours, a chill and death after 

 twenty-four hours. With larger doses the temperature falls after 

 from one-half to one hour and death results within from twelve to 

 twenty hours. Smaller doses cause a less marked reaction and the 

 animal recovers within twenty-four hours. Rabbits succumb only 

 after repeated subcutaneous injections. The substance can be ex- 

 tracted from the liver, muscles, kidneys, and urine of the poisoned 

 animals. It can also be obtained from cultures of a cholera infantum 

 germ. It is quite certain that this substance is an artificial product. 



Tetanus. — Brieger has obtained from cultures of the tetanus germ 

 four poisonous substances. The first, tetanin, which rapidly decom- 

 poses in acid, but is stable in alkaline solutions, produces tetanus in 

 mice when injected in only a few milligrammes. The second, teta- 

 noxin, produces first tremor, then paralysis, followed by severe con- 

 vulsions. The third, to which no name has been given, causes teta- 

 nus, accompanied by free flow of the saliva and tears. The fourth, 

 spasmotoxin, induces clonic and tonic convulsions. The same in- 

 vestigator isolated tetanin from the amputated arm of a man with 

 this disease. More recent researches lead us to attach but little 

 importance to the crystalline bodies discovered by Brieger, and it is 

 highly probable that the crystals with which he worked were not of 

 themselves poisonous, but were mixed with small quantities of the 

 toxin. 



In their researches on toxalbumins Brieger and Frankel obtained 

 from cultures of the tetanus germ in bouillon containing grape sugar, 

 a substance soluble in water, which when injected subcutaneously 

 in guinea-pigs caused tetanus to appear after about four days, and 

 led to a fatal termination. 



Later, Brieger and Cohn prepared tetanus poison from cultures of 

 the bacillus in veal broth containing one per cent, of pepton, and 

 one-half per cent, of common salt. These cultures were rendered 

 germ-free by filtration through porcelain, and treated with ammo- 

 nium sulphate to supersaturation. This throws the poison out of 

 solution and it floats on the surface, from which it is removed by a 

 platinum spatula. This crude poison, when dried in vacuo, is found 

 to contain 6.5 per cent, of ammonium sulphate. Of the filtered cul- 

 ture 0.00005 c.c. suffices to kill mice. From one liter of the culture 

 one gram of the dried substance was obtained, and of this 0.000,000,1 

 gram killed a mouse with the typical symptoms of tetanus. This 

 crude product contains, besides the poison, albumins, pepton, amido- 



