22,4 Goodpasture: Poisonous Constituent in Cholera 441 



apparently from morphia intoxication as it did not recover 

 completely from anaesthesia. 



It was evident from this experiment that in 100 cubic centi- 

 meters of fluid thus treated only a small quantity of toxic 

 material was present. In the remaining experiments a light 

 flocculent precipitate was removed from the intestinal fluid by 

 the addition of alcohol, and this fraction redissolved and in- 

 jected into dogs was found to be toxic. 



EXPERIMENT 2 



Two hundred cubic centimeters of fluid, prepared from the 

 sample used in the above experiment, exactly in the same way, 

 was strongly acidified with 2 cubic centimeters of glacial acetic, 

 and an equal volume of 95 per cent alcohol added. An abundant 

 flocculent precipitate resulted which was separated by centrif ug- 

 ing, washed twice with 95 per cent alcohol, once with absolute 

 alcohol, and once with ether, pressed between filter paper, and 

 dried; weight 0.3 gram. Most of this material dissolved on 

 warming in 20 cubic centimeters of water made alkaline with 

 sodium carbonate. A small undissolved residue was centrifuged 

 off. The supernatant fluid was injected intravenously into a 

 dog weighing 7 kilograms, under light ether anaesthesia, at 

 10.30 a. m. At 11 a. m. the animal had recovered from the 

 anaesthesia. He was vomiting bile-stained material; respiration 

 was deep and labored, pulse weak. Salivation was noticeable 

 and a little later tenesmus with the passage per rectum of a 

 small amount of mucus-containing fluid. At 12 noon his con- 

 dition was improved, and recovery followed. 



The substance recovered by alcoholic precipitation evidently 

 contained toxic material, but this fluid removed from the intes- 

 tine post mortem contained much more proteid than the watery 

 discharges from patients early in the disease, and the alcoholic 

 precipitation recovered from the latter proved to be more toxic. 



EXPERIMENT 3 



From 500 cubic centimeters of clear amber colored stool 

 with flakes of mucus on the surface, from a typical case of 

 cholera one day after onset, 0.355 gram of dried alcoholic precip- 

 itate was recovered. The original fluid stool was strongly 

 alkaline. It was acidified weakly to litmus with acetic acid, 

 and boiled. A light flocculent precipitate was removed by 

 centrif uging. To the remaining supernatant fluid (500 cubic 

 centimeters) two volumes of 95 per cent alcohol were added. 



