In 1898 Kiinnemann (Le) isolated the same species from soil and 
a variety from horse-dung and straw. 
By accumulation experiments, logically carried out, L have succeeded 
in obtaining this bacillus from soil, canalwater, sewage-water and 
horse-dung. 
The following experiment always led practically to a pure culture 
from canalwater : 
A bottle of about 200 Cem. is partly filled with fresh canalwater 
with addition of 2°/, calcium-tartrate, 2°/, KNO, and 0,05°/, K,HPO, 9, 
computed after the whole capacity. Then the bottle is filled up to 
the neck with canalwater and the stop is loosely put in, so that 
a little water is pressed from the bottle. In this way it is filled 
without a single bubble of air and, after shaking, put in a thermostat 
of 25° of 28°. The calcium-tartrate solves at this temperature for 
only 1°/,, so that this salt remains for a great part at the bottom. 
Commonly already after one day a feeble production of gas is to be 
observed, issuing from the non-solved calcium-tartrate at this bottom. 
The process gets into full course after three or four, sometimes only 
after five days. So much gas thereby is produced that a coarse, 
slimy scum originates at the surface and a great quantity of the 
liquid is pressed out of the bottle. The gas containing only 
nitrogen and carbondioxyd, the culture remains anaerobic. The liquid 
grows turbid by the growth of the bacteria and the fine, cristalline 
calcium-tartrate changes into coarsely granular calcium-carbonate. 
After a week, in consequence of the scum formation, the bottle is 
nearly half void, and after about 12 days the reaction is at an end, 
in as much, corresponding with the chosen quantity of tartrate, all 
nitrate has disappeared. 
If a vigorously growing culture is sown on broth gelatin, a mixture 
is obtained of colonies of various different species, from which 
B. stutzeri ean easily be isolated, if we are once acquainted with it. 
From such a bottle some drops are inoculated into a bottle of 
about 50 eem. capacity’), which, after sterilisation, is filled for */, 
with the following sterile culture liquid: ; 
Tap-water, 2 °/, calcium-tartrate, 2°/, KNO, and 0,05 °/, K,HPO,. 
After inoculation the bottle is quite filled up with the same liquid 
in the above described way, and after the lapse of two or three 
days, the same phenomena appear as in the first bottle. 
If now, once more, of this transport a plate culture on broth 
gelatin is made, the great diminution in the number of species is 
1) The capacity of the bottle is not indifferent, 
