546 FRANK R. LILLIE 



from the fertilizin. The same observation was made also on other 

 occasions. • 



The presence of the anti-fertihzin in the eggs could also be 

 demonstrated in extracts of fresh eggs. These were made (1) 

 by extracting in distilled water, (2) by drying eggs and grinding 

 them in sea-water, (3) by grinding fresh eggs with sand in a 

 mortar. I had previously often noticed that such extracts 

 gave decidedly atypical agglutination of the spermatozoa, as 

 compared with the action of the secretion of the eggs into sea- 

 water. Distilled water extracts were then examined to find the 

 cause for such atypical behavior. After it became clear from 

 the experiments quoted above that the eggs contained the anti- 

 fertilizin, the method of making the distilled water extracts was 

 modified by first shaking the eggs as free from jelly as possible, 

 so as to avoid the great excess of fertilizin contained in the jelly. 

 These experiments yielded the following results: 



August 22, 1913: 10:35 a.m. washed eggs were shaken six times 

 in a test tube to get rid of the jelly. The eggs were washed twice again 

 and shaken six times more; 60 per cent lost jelty; let stand. 2:15 p.m. 

 Poured off sea-water to 2 cc. and added fresh sea-water to 8 cc. and 

 divided equally in two test tubes, A and 5; allowed to settle. 2:27 p.m. 

 Poured off sea-water from settled eggs to 1 cc. in both A and B. To A 

 added 7 cc. sea-water. To B added 7 cc. distilled water. The eggs 

 were allowed to stand until they were completely plasmolyzed in B 

 and the supernatant fluid was bright red. Tests were then made within 

 an hour, or less, of the supernatant fluid in A and B, for the fertilizin, 

 with somewhat surprising results: 



Thus if only extreme dilutions were compared the conclusion would 

 be that B is 32 times stronger than A , whereas comparison of the undi- 



