Nov. 19, 1921 Hemotoxins from Parasitic Worms 401 



It may be concluded, therefore, that the hemolytic agent of Ascaris 

 lumbricoides is primarily a secretory product of the intestine and that 

 part of this substance finds its way into the body fluid where it is ap- 

 parently neutralized by blood elements that are obtained from the 

 host. 



7. EXPERIMENTS WITH DIFFERENT CHEMICAL. FRACTIONS OF ASCARIS 



I^UMBRICOIDES 



In contrast to the comparatively slight solubility of the hemolytic 

 substance of Ascaris lumbricoides in physiological salt solution is its 

 ready solubility in lipoid solvents, especially in alcohol. Equal quanti- 

 ties of powder were suspended in 5 cc. each of physiological salt solu- 

 tion, 95 per cent alcohol, ether, and acetone for 48 hours. The filtrates 

 were evaporated and redissolved in 5 cc. of physiological salt solution. 

 These extracts were then tested on a 5 per cent suspension of washed 

 rabbit red blood cells. The alcoholic extract was the most potent 

 from the point of view of hemolysis. Acetone and ether extracts were 

 about as potent as the physiological salt-solution extract. In a second 

 series of experiments in which A. lumbricoides powder was extracted 

 in the substances referred to above, the extracts were tested on sheep 

 red blood cells. In those experiments the alcoholic extract was the 

 most potent, while the physiological salt-solution extract and the ether 

 extract were the least potent. 



Further experiments with different fractions of Ascaris lumbricoides 

 were performed. Dried worm material was ground up in a mortar and 

 extracted in four volumes of ether in a flask for 48 hours at 37° C. The 

 ether was then removed from the worm material and saved and fresh 

 ether was added to the flask. This was allowed to extract for 24 hours, 

 the ether being removed at the end of that period and added to the 

 first ether extract. To the worm material fresh ether was again added, 

 and after 24 hours of extraction the mixture was filtered. The last 

 ether filtrate was practically free from any extract, A portion of the 

 ether extract was then evaporated and a brownish yellow fatty sub- 

 stance left behind. This substance had the characteristic odor of A. 

 lumbricoides. A small quantity of this substance was emulsified in 

 physiological salt solution and tested on washed rabbit blood corpuscles, 

 which it hemolyzed. A second portion of ether extract in solution was 

 shaken with an equal quantity of distilled water and allowed to remain 

 at room temperature for two hours. Two layers — ^namely, an ether 

 layer (fraction i) and a water layer (fraction 2) — were separated. The 

 ether layer (fraction i) was evaporated, and a fatty substance was left 

 behind which was hemolytic to washed sheep corpuscles. A portion 

 of this substance was redissolved in ether, and to this solution an equal 

 quantity of a solution of sodium bicarbonate was added and the mix- 

 ture was thoroughly shaken. The ether layer (fraction la) was removed 



