( 672 ) 
Fat-splitting moulds and yeasts can be accumulated in a feebly 
acid culture liquid, which, besides fat or fatty acids, contains only 
anorganie salts, with garden soil for inoculation material. They 
produce, together with endo-lipase, often also «- and g-lipase. 
In my experiments the fat-splitting power of bacteria was usually 
demonstrated by means of a method with fatcoated testtubes, which 
is carried out as follows. 
The inner side of a sterile test tube is coated with a thin layer 
of fat; now a nutrient liquid is introduced in which the bacterium 
to be examined on lipase grows well and this bacterium is inoculated 
into it. If now lipase is produced in the culture we see after two 
or three days that portion of the fat which touches the liquid grows 
white; this appears first and most obviously at the place where 
the bacterial growth is strongest. Aérobes decompose the fat first 
near the surface of the liquid, anaérobes first at the bottom of 
the tube. 
On the accompanying photographs Pl. 3 we see a series of fatted 
tubes in which various fat-splitting microbes have decomposed the 
fat. In 1 no fat-splitting bacterium is inoculated, hence the fat has 
remained unchanged; 2 and 8 contain cultures with ammonium 
chloratum as source of carbon resp. of B. Stutzeri and B. denitro- 
fluorescens non-liquefaciens ; 4 and 5 contain cultures of the same 
microbes but with kaliumnitrate as source of nitrogen; 6 contains 
a rough culture of a proteid putrefaction by inoeulation with soil; 
7 contains a rough culture of a pasteurised proteid putrefaction ; 
8, 9, 10, 11, and 12 contain cultures of various fat-splitting 
microbes isolated from soil and milk. 
Il. FAT-SPLITTING MICROBES IN THE SOIL. 
The microflora of the soil aboands in organisms which secrete 
lipolytic enzymes; a sowing of soil on a fatted plate after Eykmay, 
or dilutions in fatted tubes show this most clearly. In one gram 
of humus we not seldom count some ten thousand fat-splitting 
ferments. 
A. Acecumulations in culture media with ammonium chloratum 
as source of nitrogen. 
For the growth of bacteria, requiring beside fat as source of 
¢arbon only anorganie salts and ammonium ehloratum as source 
of nitrogen, the following culture liquid was used. 
