200 PROTEIN POISONS 



This work was not resumed until 1902, and on repeating 

 the work the alcoholic precipitate failed to manifest any 

 poisonous action, but in this instance we heated the acid 

 extract for two hours. Inasmuch as the prolonged heating 

 seemed to be the only difference in the methods of pro- 

 cedure, another trial was made in which the acid extract 

 was heated in the autoclave at 110 for exactly ten minutes. 

 The alcoholic precipitate freed from acid as before was 

 ground to a fine powder in an agate mortar. An unweighed 

 portion of this powder was dissolved in 5 c.c. of water and 

 injected intra-abdominally. When last seen that night, 

 nine hours after the injection, the breathing was difficult 

 and irregular. The animal was found dead the next morning. 

 Autopsy showed extreme subcutaneous edema over the 

 abdomen. The peritoneal cavity contained a few cubic 

 centimeters of a clear fluid, and a smaller amount of bloody 

 exudate was found in the pleural cavity. The heart was 

 in diastole and the most marked changes were found in 

 the lungs. These were greatly congested and the left upper 

 lobe seemed to be consolidated. Closer examination showed 

 portions of the lungs to be completely hepatized. Many 

 of the air cells were filled with exudate and blood corpuscles. 

 The kidneys were highly congested and the liver seemed 

 pale and friable. Further experiments with weighed por- 

 tions of the powder showed the minimum fatal dose for 

 a guinea-pig when given intra-abdominally to be about 

 50 mg. Smaller doses down to 15 mg. made the animals 

 very sick, but failed to kill. 



The poisonous group obtained from the anthrax protein 

 by cleavage with 1 per cent, sulphuric acid is destroyed, 

 at least greatly weakened, by prolonged boiling in aqueous 

 solution. 



The cellular substance of the anthrax bacillus, prepared 

 by our method, is the least toxic of all the bacterial proteins 

 with which we have worked. This is true whether the cell 

 protein is derived from a pathogenic or a non-pathogenic 

 organism. It requires not less than JO mg. of the cell 

 substance, after extraction with ohol and ether, to 



