9206 TRANSACTIONS LIVERPOOL BIOLOGICAL SOCIETY. 
sloped nutrient agar in tubes. After 24 hours at AQ? 
these secondary sub-cultures were again sub-cultured in 
the following sugar media:—Glucose, lactose, mannose, 
sucrose, dulcite, inulin, and adonite, all containing 
litmus. Litmus milk cultures were also made, and Voges’ 
and Proskauer’s reaction was tested for in the glucose 
cultures. All were incubated for 48 hours at 42° C. At 
first I examined the sugar cultures for motility of the 
bacilli, but later on abandoned this character as useless. 
In each case 1 e.c. of the cockle liquid in each flask 
was incubated anaerobically in litmus milk previously 
heated to nearly 100 ¢.c. and then cooled rapidly. By the 
‘“ Hnteritidis reaction ” is meant the clotting of the milk 
after 24 hours’ strictly anaerobic incubation; the appear- 
ance of acid, and extensive disruption of the clot by gas 
formation. No further cultures and no inoculations were 
made, 
I think it essential to give the details of the analyses, 
since future examination of some of the cockle beds may 
prove to be of great importance, and if such analyses are 
made and their results compared with those recorded here 
the comparison will hold good only if the methods 
employed are identical. There is, unhappily, no doubt 
that different bacteriological methods of enumeration of 
Bacillus coli in shell-fish may give strikingly discrepant 
results, and whether or not the method adopted above is 
the best, comparison analyses should be made by means 
of it. 
I give now the details of the analyses, stating in each 
case the results of the primary cultures, as well as those 
of the tertiary sub-cultures. 
lL. The Formby <cockile ene 
The samples were collected one hour before low 
water: (a) from Formby shore near a gutter on the sands 
and W.S.W. from the old Victoria Beach mark; and 
