E. E. Atkin and A. Bacot 
48y 
was carefully cleaned and a number of larvae were washed in five changes 
of tap water, by thorough agitation and violent currents set up by a 
water-jet; there is little danger of the larvae being washed out of a 
deep pan, as they invariably keep at the bottom when the water is 
agitated. They were then left 24 hours, so that they might void the 
contents of their guts, and then again washed in two or three changes 
of water. After this preliminary treatment they were put into the 
clean pan and fed exclusively on broth cultures of Bacillus coli. As 
a result of this procedure the percentage of breakdowns in the sterility 
of the experimental tubes was much reduced. 
One or two precautions with regard to pipetting the eggs are also 
advisable. Floating eggs should be avoided, and single detached eggs 
alone should be taken up. When two or more eggs remain attached 
after immersion in the lysol there is a probability that at the point of 
attachment a portion of the surfaces will not have been completely 
sterilized. The reason for avoiding floating eggs is also the danger of 
incomplete sterilization. In order to get rid of eggs attached to each 
other, the eggs may be sifted through motor veil gauze or any other 
very fine gauze. 
The whole of the experiments are placed as an appendix, references 
to .them only being given in the text. 
Section A. Eggs. 
Effect of contaminated water. Unsterile conditions. 
Experiments I and II, already referred to on page 483, show clearly 
the powerful action of water charged with organic matter and living 
organisms on dormant eggs which failed to hatch in clean water. 
Effect of alkaline solutions—under unsterile and sterile conditions. 
Experiments III and IV. It was suggested by one of our colleagues 
that the results which followed the addition of water or other media 
charged with bacteria or yeasts might be due to the ammonia content. 
Aggramonte showed (1902) that the presence of lye from wood ashes 
stimulated the early hatching of the eggs. Two trials were carried out, 
one under unsterile and the other under sterile conditions. From 
these it seems quite clear that, although a few eggs may respond to 
the presence of alkaline solutions in either tap or distilled water, the 
efiect is very feeble in comparison with that due to either contaminated 
water or a broth culture of B. coli. 
