4i8 



E. E. JUST. 



offset the possibility that the failure of the shed eggs to cleave is 

 due to the presence of blood with the sperm. 



(c~) July 27. Samples of shed eggs from 14 females insemi- 

 nated in turn. Very high amount of inhibition present in each of 

 Nos. i to u, inclusive. Membrane separation in these eggs as 

 follows : 



Remainder of uninseminated eggs of Nos. i to n, inclusive, 

 put in one dish. "Washed in five changes of 250 c.c. of sea-water. 

 Inseminated. 13 per cent, cleavage. 



Ovaries from these same females chopped up separately in sea- 

 water; eggs strained and collected. Portion of each inseminated. 

 Average fertilization 95 per cent. Remaining eggs added and 

 inseminated in one dish. Fertilization 97 per cent. 



Eggs from Nos. 12, 13, 14, added, inseminated. Fertilization 

 close to 100 per cent. 



These uninseminated shed eggs gave high agglutinin test. I 

 thought it worth while to add the eggs from several females in 

 order to increase the amount of sperm agglutinin (fertilizin) ; 

 this, however, failed to improve results. These eggs do not fail 

 to develop because of lack of fertilizin or of failure of sperm 

 entry. They are loaded with blood inhibitor and this suspends the 

 fertilization-reaction. 



(d} July 30, 9:50 A.M. 9 females opened, gently drained of 

 blood. Each lot of shed eggs then in finger bowls of 200 c.c. of 

 sea-water. Portions of these eggs removed and inseminated at 

 10:00 A.M., 10:28 A.M., 10:43 A.M., and 11:25 A.M., with 

 the following results : 



1 Not sufficient eggs left to make count. 



