576 Dale. — Observations on Gymnoascaceae. 
stained in various ways. The best results were obtained with 
Flemming’s triple-stain-safranin, gentian violet and orange G, 
and with toluidine blue and eosin. The latter stain is some- 
what uncertain, but when successful the results are very good. 
The eosin stains the nucleoli red, while the toluidine blue 
stains the protoplasm blue. 
A very useful stain for these Fungi is brazilin 1 , which dif- 
ferentiates the nuclei very clearly. Its special advantages are 
that its effects are very certain, and there is no over-staining. 
The results seem to be equally good, whether the material is 
stained before or after cutting. 
I. The Life-History of Gymnoascus Reessii. 
The original material consisted of little brick-red balls, made 
up of thick-walled septate hyphae, freely branching and anasto- 
mosing, and enclosing a mass of ripe ascospores, spherical in 
form and of a pale brown colour. These spores were for the 
most part isolated, but some were still contained in the 
spherical asci (PI. XXVII, Fig. 1). 
The thick-walled hyphae branch in a peculiar manner, the 
branches arising almost at right angles to the axis which bears 
them. Thus anastomosis is facilitated, and also the dense 
growth which results in the spherical mass of hyphae sur- 
rounding the groups of asci. The branches are said by Fischer 2 
to be covered with ‘short, straight, or slightly bent spines, 
10-15 j ul long.’ Both in the original material and in the 
cultures subsequently made from it, the ends of the hyphae 
were blunt (Fig. 1 a). The hyphae were either empty or 
contained a greater or less amount of protoplasm. None of 
the asci was attached to any hyphae. 
The ascospores readily germinated in various nutritive 
media. Those chiefly used were beer-wort, or horse-dung 
extract, made up with 10 per cent, or 15 per cent, of gelatine. 
Colonies were afterwards transferred either to sterilized horse- 
1 Hickson, Q. J. M. S., vol. 44, p. 469 (1901). 
2 Engler und Prantl, loc. eit., p. 294. 
