40 PROTEIN POISONS 



supernatant fluid, the latter being somewhat opalescent. 

 Guinea-pigs were treated with both portions, and all died. 

 This demonstrates that superheated steam disrupts the 

 bacterial cells, but does not destroy the intracellular poison. 

 Heating the cellular substance of the colon bacillus in 

 physiological salt solution to 140 in the autoclave does 

 not destroy or even weaken the poison. The cell substance 

 used in this experiment had been prepared six years pre- 

 viously. One gram was thoroughly mixed with 100 c.c. of 

 salt solution. Two cubic centimeters of this mixture, con- 

 taining 20 mg. of the cell substance, killed guinea-pigs of 

 300 grams' weight when injected intra-abdominally, and 

 0.5 c.c. or 5 mg. made the animals very sick. The emulsion 

 was then heated in the autoclave to 140 and held at this 

 temperature for ten minutes. One cubic centimeter of 

 this heated emulsion, containing 10 mg. of the cell sub- 

 stance, killed guinea-pigs of 300 grams' weight, and 0.1 c.c. 

 or 1 mg. made the animals sick. Animals treated with the 

 heated emulsion died more promptly and from smaller 

 doses than those treated with the unheated preparation. 

 Evidently the heating prepared the cell substance so that 

 it was more promptly split up by the ferments of the body. 

 When heated to this temperature .a part of the poison 

 passes into solution. This was shown by filtering the 

 heated emulsion through hard paper. The filtrate was 

 clear, slightly acid to litmus, gave the biuret, Millon, and 

 -naphthol tests, and killed guinea-pigs of 300 grams' 

 weight. The animals that had the unheated suspension 

 showed marked peritoneal inflammation, with bloody exu- 

 date. In those having the larger doses (40 mg. of the cell 

 substance) the inflammatory condition extended to the 

 muscular walls of the abdomen. In those that had the 

 heated suspension the inflammatory condition was much 

 less marked, there being only a slight serous exudate, less 

 and less stained with blood as the amount of the cell sub- 

 stance injected decreased. In those killed with the filtrate 

 there was no evidence of peritoneal inflammation. This 

 furnishes a beautiful illustration of the nature of inflam- 



