36 BOWMAN. 



the intestine were negative. However, the organism was recovered from 

 a stool passed three days after feeding the bacterial suspension. 



After the organism had been grown for one month its pathogenicity 

 was again tested. Comparatively large doses failed to kill either guinea 

 pigs or rabbits. It is evident fro, a this that Bacillus "S" rapidly loses 

 its pathogenicity. The original fatal dose was again obtained after 

 passage through guinea pigs. 



The filtrate from living cultures. — Two flasks, each containing 150 

 cubic centimeters of sterile bouillon, one being 1.5 per cent alkaline, the 

 other two per cent acid, were inoculated and allowed to grow at 37° C. 

 for twenty-four hours. The cultures were then each passed through 

 a Berkefeld filter, and the filtrate injected in large and small amounts 

 intraperitoneally into guinea pigs, and intravenously into rabbits. The 

 result was negative. The cultures after growing for seven days in 

 alkaline and acid bouillon respectively were filtered and the filtrate 

 inoculated into rabbits and guinea pigs with negative results. 



A search was then made for an endotoxin. Six complete slants of 

 agar in tubes were inoculated and the organism allowed to grow twenty- 

 four hours, it was then suspended in salt solution and placed in a shaking 

 machine for eighteen hours. The filtrate was injected into guinea pigs 

 -and rabbits, without result. 



Suspension of "killed cultures. — Suspension of 24-hour growths of the 

 bacilli were placed in an incubator at 60° C. for one hour, and plates 

 were then made demonstrating the sterility of the suspension. Two 

 guinea pigs were inoculated each with 2 cubic centimeters of the suspen- 

 sion ; one rabbit received 2 cubic centimeters intravenously. After 

 twenty-four hours the guinea pigs were both dead, the rabbit still alive. 

 At autopsy it was demonstrated that the peritioneal cavity of each guinea 

 pig contained a large amount of sero-purulent fluid. However, smears 

 and cultures from this were negative. At the end of forty-eight hours 

 the plates made from the killed suspension were examined. They were 

 sterile. 



Agglutination tests. — As all four cases occurred in private practice 

 it was not possible to obtain blood from each patient. Blood in very 

 small amount was obtained from Case I on a slide, and although accurate 

 quantitative tests could not be made, those which were conducted were 

 very suggestive. 



The blood from Case I was diluted with salt solution 1 to 50 and 

 hanging drop preparations made with controls in each case. Table III 

 shows the result. 



