22,4 Goodpasture: Poisonous Constituent in Cholera 443 



The flasks were then placed in the ice box at 10° C. for several 

 weeks until the material in suspension had sedimented, leaving 

 a clear supernatant fluid. This was passed through a Mandler 

 filter. The filtrate was water clear and amber colored. Five 

 hundred cubic centimeters were made acid by the addition of 2 

 cubic centimeters of glacial acetic and two volumes of 95 per cent 

 alcohol were added. A light flocculent precipitate separated out, 

 estimated at about 0.2 gram. This was removed by sedimen- 

 tation and centrifuging, and washed once with 95 per cent 

 alcohol. The alcohol was decanted and the precipitate com- 

 pletely dissolved at room temperature in 20 cubic centimeters of 

 water made slightly alkaline with sodium carbonate. This 

 solution injected intravenously into a dog weighing 6 kilograms 

 resulted in vomiting, watery diarrhoea, collapse, and death within 

 one hour. Autopsy showed splanchnic congestion and intense 

 reddening of duodenal mucosa. 



The precipitate from an additional 500 cubic centimeters 

 treated in the same way was injected intraperitoneally at 10 a. 

 m. into a dog weighing 5.5 kilograms. There followed almost 

 immediate collapse, fall in blood pressure, labored respiration, 

 urination, and tenesmus. At 11.30 a. m. collapse continued. 

 Passing watery discharges from bowel. Pulse could not be felt 

 at 12 noon. Copious fluid fsecal discharges. Vomited blood- 

 stained fluid. Convulsions. 12.30 p. m., dead. At autopsy an 

 intense congestion of the intestinal mucosa was found, although 

 the peritoneal surfaces were pale and smooth. Large, irregular, 

 brick-red splotches of congestion were distributed throughout 

 the mucosa of the small bowel. Mucosa of colon congested. 



Since the fluid stools from cholera patients contain practically 

 no proteid precipitable by heat and acetic acid, the above method 

 of preparing the alcohol precipitate from water-clear fluid which 

 has passed through a diatomaceous filter is much the best. Even 

 an initial heating to sterilize the fluid is not absolutely necessary ; 

 initial heating may destroy some of its toxicity but was uni- 

 formly done in these experiments as a precautionary measure. 

 It has been found more difficult to dissolve the dried precipitate 

 than that washed once with alcohol, and the washed preparations 

 appear to be more toxic. 



No attempt has been made to purify the poison contained in 

 the faecal precipitate, and it is undoubtedly brought out of 

 solution along with other substances. Precipitation from an 

 acid solution is much easier than with the natural alkaline 

 reaction. Four volumes of alcohol or more are necessary with 



