534 FRANK R. LILLIE 



TABLE 2— Continued 



■WASHINGS 



TESTS 



BULK OF EGGS I TIME OF CHANGE 



1913 

 August 17 



34 12/1.5 cc. 1/20— 5 sec. 0.325 cc. 9:15 a.m. 



35 On stirring up, the eggs disintegrate, and the fluid becomes highly 



colored; the agglutinating substance is absolutely neutralized. Test 

 1/1 negative, but the fluid is highly chemotactic and activating. 



In order to determine the role of the jelly in charging the sea- 

 water with fertilizin, I killed some eggs by heating them for 5 

 minutes to 60° C. (August 15, 1913,) and submitted them to a 

 series of 22 washings in 3 days, by which time the jelly was nearly 

 all dissolved away : the agglutinating power decreased from 800 

 at first to 1 at the end. The agglutinating substance could be 

 detected as long as any jelly remained. 



On the other hand, eggs may be entirely deprived of jelly by 

 shaking, and such eggs will continue to produce fertilizin as long 

 as eggs with jelly, though in lesser quantities at first. This matter 

 is so fundamental that I include another long table giving a com- 

 parison of eggs deprived of jelly by shaking and those with jelly 

 (table 3). 



August 21, 1913: In this experiment ripe eggs of Arbacia were 

 washed once and divided in three equal lots, A, B, and C, in 10 cc. sea- 

 water in graduated cylinders. They were allowed to settle, and at 

 9:20 A.M. the bulk of eggs was 1.8 cc. in A, 2 cc. in B, and 2 cc. in C. 

 Thus there were about 10 per cent fewer eggs in A than in B and C. 

 At 9 :23 A was transferred to a test-tube and given 6 vigorous shakes to 

 get rid of the jelly. A sample was then examined in India ink and it 

 was found that 90 per cent of the eggs were entirely freed from the jelly; 

 in the remainder it was usually reduced to a thin layer. A was then 

 returned to its cylinder in 10 cc. sea-water and allowed to settle. As 

 the eggs settled in A it could be seen that the fluid was only sHghtly 

 colored; so that it was certain that not many eggs were injured in the 

 shaking.^ At 9:40 a.m. the bulk of eggs was as follows: A, 0.6 cc; 

 B, 1.6 cc; C, 1.6 cc. As care was taken to avoid loss of eggs in A the 



- It should be noted that in such an experiment, it is extremely important not 

 to shake too much, for if the eggs themselves are injured so that the color escapes, 

 a complicating factor comes in considered beyond (see section 6, p. 544). 



