A NEW SPECIES OF ALCOHOL FORMING BACTERIA 241 



Dissolved in 2000 cc. of the following salt solution: 



dis'OTjVed in 

 5000 cc. 



WATER 



grarns 



Potassium phosphate 5 



Magnesium sulphate 1.0 



Calcium chloric! 0.5 



The solution was divided into one liter portions sterilized 

 by the intermittent method, and inoculated with a pure culture 

 of Bacillus saccharalis. After an incubation period of two weeks, 

 an alcohol determination was made. The liquor was evaporated 

 down to about a third of the original volume, until the distil- 

 late gave only a faint reaction with iodoform. After repeated 

 distillations, the specific gravity of the 50 cc. portion was found 

 to be 0.99744 which corresponds to 1.707 per cent of alcohol 

 by volume. By dehydrating a small portion with fused car- 

 bonate of potash a solution was obtained which distilled at 79° 

 to 80°C. showing it to be ethyl alcohol. 



The residue was tested for acids and acetic acid was found 

 to be present, using the ethyl acetate test, A mannite solu- 

 tion made up according to the same formula as the glycerin 

 solution, except that 3 per cent of mannite was substituted 

 for an equal weight of glycerine, was next tried. It yielded 

 50 cc. of a distillate with a specific gravity of 0.99836 correspond- 

 ing to an alcohol per cent of 0.55 by volume. The presence of 

 acetic acid was also detected in the residue. 



Thinking that the low yields of alcohol in the two cases was 

 due to the small quantity of assimilable nitrogen in the solution, 

 3 per cent of glycerin was added to plain bouillon, and the 

 flask sterilized and inoculated as before. In this case the solu- 

 tion yielded 50 cc. of distillate of a specific gravity of 0.9864, 

 which corresponds to an alcohol per cent of 4.895, which was 

 much higher than in the previous experiment. The higher 

 yield in the latter case indicated that there was a lack of nitro- 

 gen in the solution previously used. Frankland and Fox in 

 their experiment with B. ethaceticus obtained a yield of 11.41 

 grams of alcohol from 60 grams of glycerin. In the experiments 

 of the above investigators it was found that B. ethaceticus formed 



