Institute, puSa, for lsif-is 129 



number into the steeping vat. In the 1917 experiments it 

 was frequently found that whereas the uninoculated 

 khazana water might contain very few hydrolyzers yet the 

 corresponding fermented liquor contained an abundance of 

 these bacteria; this implied the introduction of the latter 

 by the plant, and it may be asked how manufacture could 

 be benefited by purification of the khazana water, if the 

 plant itself brings in so large a number of bacteria. There 

 is reason to think, however, that this difficulty is not so 

 serious a one as at first sight may appear, first because the 

 bacteria generally associated with the plant do not seem to 

 include as a rule any large number of deleterious organisms, 

 and secondly because these bacteria, if they find the water 

 already in possession of others, as would be the case with 

 successfully inoculated khazana water, do not have time to 

 exercise much influence upon the character of the fermen- 

 tation. For this same-reason beneficial plant bacteria must 

 frequently be prevented from carrying out normal hydro- 

 lysis by the presence of unduly large numbers of detri- 

 mental organisms in the khazana water. The problem 

 therefore is how far is it practicable to provide fairly clean 

 water for steeping, and how to produce a satisfactory artifi- 

 cial inoculation sufficiently vigorous to overcome the des- 

 tructive action of detrimental bacteria already present. 



A great deal of experimental work was done to deter- 

 mine the best way of making a " mass " culture, i.e., a pure 

 culture of the specific bacterium sufficient in quantity to 

 stand distribution through the 6,000 to 10,000 gallons of 

 water in the steeping vat without undergoing too high 

 dilution. It was found possible to make a simple culture 

 medium of ammAiium sulphate, superphosphate, wood ash 

 and sugar, and to activate fermentation by the use of about 

 one gallon of this culture, after 6 hours' incubation, in each 

 1,000 gallons in the steeping vat. In many cases, how- 

 ever, growth of the artificial inoculum appeared to be inter- 

 fered with, probably owing to excessive competition with 

 other bacteria and in some instances to failure to make the 

 proper adjustments between the temperature of the mass 



