168 
A. E. Lechmere. 
The pieces bearing mycelia were isolated and well washed in 
fresh water and transferred to Petri dishes containing distilled 
water. 
On examination of the series of mycelia isolated in this manner, 
two distinct species were found to be present; each of these was 
then isolated into pure culture by the following method. A small 
piece of the mycelium was teased off with fine needles and trans¬ 
ferred to meat extract gelatine in Petri dishes. Full details of this 
method are given in the other paper. 
In each case the fungus grew very rapidly on this medium, but 
at the same time produced very vigorous liquefaction, which enabled 
the protozoa and bacteria to spread to the edge, and so rendered 
isolation of hyphae free from bacteria difficult. 
To overcome this point, the plates were tilted at an angle of 
60", the direction of the vertical being marked on the back of the 
dish by an arrow drawn in wax so that the plate could always be 
replaced in the same position. In this way all the bacteria, etc., 
were carried to the lower part of the plate culture, leaving a fringe 
of hyphae perfectly free from organisms growing towards the upper 
portion. Other plates were inoculated with small fragments of 
hyphae taken from the extreme upper edge of a two days’ growth, 
and very soon the cultures were obtained free from organisms. 
Growth on the gelatine medium is always entirely vegetative, 
so, in order to follow the life-history of these two species, a series 
of cultures in each case was made on white of egg in distilled water, 
sterilized Petri dishes being used throughout. 
As a result of the previous investigation, white of egg was found 
to be by far the most convenient medium with which to work. 
The stock of albumen is easily prepared by placing small slices 
of the white of a hard-boiled egg in test-tubes containing water. 
These tubes can then be sterilized in an autoclave at 120°C for 
twenty minutes, and so preserved for future use. For the purpose 
of the cultures, small pieces of albumen can be removed from these 
slices by means of a thick platinum needle and transferred to the 
Petri dishes containing distilled water. 
After two to three days’ growth on the albumen, a fairly dense 
fringe of hyphae is developed, and formation of sporocystscommences, 
at this stage the water is changed every other day to keep the 
cultures fresh. 
After a fairly vigorous mycelium had been developed several 
cultures were allowed to remain without changing the water, to 
