( 669 ) 
in those containing an easily assimilable source of nitrogen, suchr as 
peptone or proteids, after inoculation with garden soil. Addition of 
calcium nitrate to this culture medium proved very favourable for 
the process. 
In his experiments with the pure culture of a fat-splitting bacterium 
in broth, caleium nitrate and 4.424 ers. of fat, after 35 days’ culti- 
vation at room temperature, more than the half was split and 
+ 0.7 er. assimilated; after a cultivation of more than a year, 
however, nearly all the added fat was split, more than the half 
being assimilated. 
The fatty acids and the glycerin formed from the fats are oxydised 
without intermediate produets to carbonic acid and water. 
BecHHoLp') demonstrated that fats and soaps disappear in the 
sewage mud of the installations for the purification of water at 
Stettin, by bacterial action. 
EYKMAN *) gives in a treatise on fat-splitting bacteria, beside other 
facts, a simple and nice method to demonstrate lipase secreted by 
micro-organisms. This method is based on the splitting action 
of diffusing lipase, produced by bacterial streaks on agar or 
gelatin, on a thin fat layer at the underside of this substratum. The 
fat, decomposed under the bacterial mass has the appearance of 
an opaque white stripe, very distinct from the non-decomposed fat. 
According to Eykman the following bacteria split fat: B. pyocyaneum, 
Staphylococcus pyogenes aureus, B. prodigiosum, B. indicum, B. ruber, 
B. fluorescens liquefaciens. 
Raun*) has studied fat-splitting by microbes on a culture medium 
of the following composition. In an inclined Erienmeyer flask some 
fat is melted. After cooling the flask is put upright and a thin 
layer of an anorganic culture liquid is filled into it. 
After inoculation with garden soil a good growth of moulds and 
bacteria results, which, after transferenve to a similar flask soon 
recommences. 
Four fat-splitting moulds and two fat-splitting bacteria were isolated 
on culture plates of an anorganie nutrient liquid soliditied with agar 
and containing tributyrin in finely divided state. On these plates a 
clear field appears around the colonies of fat-splitting microbes, in 
consequence of the splitting action of the diffusing lipase on the 
tributyrin, and of the solution of the products formed in the medium, 
1) Zeitschr. f. Angew. Chem. 1898 S, 849 cit. 4). 
*) Eighth Dutch Phys. and Med. congress 1901 p. 171, 
5) Ceniralblatt f. Bakt. 2 Abt 1906 Bd. 15 S. 422, 
