(583 ) 
For the continuation of the culture by inoculating slight quan- 
tities of material of a rough fermentation into the same nutrient 
liquid, two precautions should be taken. First, the inoculation should 
be done into the medium, freed from air by boiling, the bottle 
being entirely filled with the hot liquid, so that on cooling no air 
can dissolve, Second, an acidity of less than 7 proves not suffi- 
cient, hence this should be 8 or 10 ec, as otherwise the lactic acid 
ferments might prevail and supplant the sarcine. 
From the necessity of expelling the air we see that the fermen- 
tation sarcine undoubtedly belongs to the ordinary anaerobics, which, 
considering the success of the rough accumulation experiment with 
aeration, might perhaps not have been expected ; but the fact holds good 
in the same way for the butyric acid ferment, generally accepted as 
an obligative anaerobic, so that, also with respect to the fermentation 
sarcine, there should be spoken of ‘‘microaerophily.” Further exami- 
nation shows that in deep test-tubes with maltwort-agar, very easily 
pure cultures may be obtained, whereby the sareine is recog- 
nisable by the obvious size and the remarkably rapid development 
of its colonies. On the other hand, on maltwort, or broth-bouillon- 
glucose-agar-plates with or without acid at 37° C., with access 
of air, no growth at all of the sarcine takes place, as might 
be expected. Of course the packages can also be seen on the 
plates without growing and be removed in a pure condition. When 
we make use of little acid for the rough accumulation, colonies of 
lactic acid ferments, belonging to the physiological genus Lacto- 
bacillus, will develop on the plates at the air, which can grow 
as well with as without air, but whose other life conditions corre- 
spond to those of the sarcine. In this case the experiment shows at 
the same time tbat everywhere in garden soil real lactic acid fer- 
ments are present, whereof the proof had not been given until now. 
When using much acid, for example 10 ce. or more normal 
acid per 100 ce. of culture fluid, through which the vital functions 
of the sarcine, such as rapidity of growth and the faculty of assimilating 
oxygen, are lessened, certain alcohol ferments, proper to garden soil, 
come to development, but they can, together with some of the other 
impurifications of the rough accumulations, as moulds, JZucor and 
Oidium, be checked and expelled by exclusion of air, hence, by 
culture in closed bottles. To this end however, it is necessary to render 
the conditions for the sarcine as favorable as possible and not allow 
a temperature below 37° C. 
The staying out of the butyric acid fermentation (caused by Granu- 
lobacter saccharobutyricum), which so readily originates with exclusion 
