16 E. E. JUST. 



1. July 2, 10:00 A.M.: Eggs from 4 females washed 5 times in 250 c.c. of sea- 

 water at each washing. 2 c.c. of eggs after the fifth washing. At 12:00 M. 210 

 c.c. of supernatant water withdrawn and filtered, 210 c.c. of sea-water being added 

 to the eggs. A sample of this removed at 1:30 P.M. on testing gave at 1/1,600 

 dilution a 10 second reaction. This is equal to morning tests. Washed 5 times 

 during the afternoon by removing 210, 217, 203, 190, and 205 c.c. of water and 

 adding in each case the quantity withdrawn. Tested at 8:30 P.M., supernatant 

 sea-water gave an 8-second reaction at 1/1,600. July 3, 9:00 A.M. 210 c.c. of 

 supernatant sea-water removed, 210 c.c. added. Tested during the day to 1:30 

 P.M. negative. 



2. July 3, 9:10: Ovaries of 5 females in sea-water strained. Sea-water added 

 to loo-c.c. mark. Washings (75 c.c. of water removed each time) as follows: 

 9:30, 9:34, 9:38, 10:05, 10:12 during the morning and at 1:50, 2:30 and 2:38 during 

 the afternoon. Tested at 9:40, 10:50 and 1:30 gave 10 second reactions at i/ioo. 

 At 2:40, 8-second reaction on 1/50 dilution. 2:55 P.M. 75 c.c. of supernatant 

 water removed, 75 c.c. added. 5 P.M., reaction faintly positive at 1/50. July 4, 

 10:30 A.M. 75 c.c. removed, 75 c.c. added, n :io A.M. negative when tested. 



2. Source of Fertilizin. 



In Arbacia only mature eggs produce fertilizin; immature eggs 

 are incapable of secreting the agglutinating substance; other 

 tissues do not elaborate it, nor does it occur -in the perivisceral 

 fluid (Lillie, '13, '14). In Echinarachnius mature eggs alone 

 with or without jelly yield fertilizin to the sea-water. Neither 

 the immature egg nor the body-fluid is capable of charging sea- 

 water with sperm agglutinin. 



(a) Comparison of Ripe Eggs With and Without Jelly. Though 

 the jelly is loaded with fertilizin, it does not form the fertilizin 

 for jelly-free eggs are capable of charging the sea-water with the 

 sperm agglutinating substance. The jelly hulls enclosing 

 Echinarachnius eggs may be removed by shaking or by treat- 

 ment with acid. The method of removing the jelly by shaking 

 demands care since agitation may cause injury to the eggs. If 

 shaking brings about the binding of fertilizin we should get a 

 cessation of fertilizin production. This should not be attributed 

 to mere loss of jelly, though it might be so interpreted by some. 

 Again, it might be argued that the use of acid to remove the jelly 

 might injure the egg; this objection could be raised only in the 

 event that fertilizin is no longer produced by the egg after acid 

 treatment, which is at least remotely possible. Certainly, it 

 would be gratuitous to argue that if the fertilizin continues to 

 be produced after acid treatment the acid carries the fertilizin 

 from jelly to egg! In short, after shaking or after acid treatment 



