145 
10). The ordinary reactions—solubility in alcohol, See Birt ag - 
mic acid, staining with alkanet, etc.—show beyond doubt t eir : y 
character. They vary in size from spheroids, 7-8 to 10-12 w= in dila- 
meter, to ovoids, 14-15 by 10-12 
It wae thus lente foe observation in hanging-drop 
cultures, that the yeast gives rise to the hyphe which swell into the 
chlamydospores, every stage in the development of which was followed. 
Attempts were then made to get the chlamydospores to germinate, 
but although it was possible to isolate them in hanging drops in spite 
of the difficulty of separating all yeast-cells—which gave trouble by 
growing first and over-crowding the chlamydospores—I found no trace 
of germination of the latter in the first state. Nor was any success 
obtained after heating the wort-gelatine containing the spores to 55°C 
for five minutes. This killed the yeast-cells, and I was able to keep 
chlamydospores under observation in hanging drops, but in no case 
was I able to obtain the slightest direct evidence of their germination. 
Of course they may have been killed also, though it does not appear 
likely. 
ee or two plates of the mixed yeast and chlamydospores in wort- 
gelatine heated as above, a few straggling colonies did appear after a 
fortnight, suggesting that one or two yeast-cells may have survived the 
heating to 55°C. 
Many other attempts were made to get the chlamydospores to ger- 
minate. Not only were they sown fresh, but also after drying, after 
freezing, and, as we have seen, after heating. Digestion in gastric 
juice was equally of no avail. 
Of the media tried, in addition to wort-gelatine, I employed wort- 
gelatine with extract of cow-dung added ; wort-agar, agar with extract 
of horse-dung; agar with decoction of willow-wood, with extract of 
rotting pine-leaves, the same with Levulose added, gelatine with beech- 
wood extract, etc. Also the dead leaves of pine, beech, oak, grass, 
fern ; the saw-dust of oak and of pine ; blocks of beech-wood and pine 
and of rotten willow, as well as the Same saturated with extract of 
willow. Pieces of mushroom were also used. Elder-pith, saturated 
with Klebs’ solution, or with extract of pine-water with or without 
xylose, in all cases without success. Nor would the chlamydospores 
germinate in any of the following carbo-hydrates, made up in 10 °/o 
solutions with yeast extract. 
Saccharose, dextrose, lactose, mannite, maltose, xylose, dextrine, 
gum, all of which were tried at 32°C, 24°C, and at ordinary tempera- 
tures, 16-18°C, as were the other more solid media. It may therefore 
be concluded that either these chlamydospores require some very 
special conditions to force them to germinate, or they are incapable 
of further growth—a very unlikely view. 
The yeast-form, on the other hand, grows readily on all the above - 
media except the following : Beech-wood gelatine, elder-pith with 
Klebs’ solution, rotten willow blocks with or without willow extract, 
mushroom, and the dead leaves and sawdust, on which it either refuses 
to grow at all or buds very sparely. 
