24 



BULLETIN 908, U. S. DEPARTMENT OF AGRICULTURE. 



confirming those found under artificial conditions. Both ammonia 

 and amines were present in appreciable amounts, the quantity of 

 amines in this instance being in excess of the ammonia. 



Laboratory Experiments with Bacillus Walpischrauschbrand and 



Bacillus B. 



To establish more firmly the conclusion that the decomposition of 

 the feed and of the fish, resulting in the condition termed belly blown, 

 is due to the presence of Bacillus Walfisclirauschbrand and Bacillus B, 

 found in great numbers in the samples of feed and swelled cans 

 examined, and to show that the presence of these bacteria is indi- 

 cated when volatile nitrogen bases are found in belly-blown fish, 

 swelled cans, etc., these bacteria were grown in pure culture upon a 

 medium containing fish protein, and the products resulting from 

 their growth observed. The fish used for cultures 24, 25, 31, and 32 

 (Table 8) were fresh Boston mackerel. Fresh-caught Potomac bass 

 were used for cultures 27, 28, 29, and 30. After removing the skin, 

 solid masses of meat were passed through a meat grinder and mixed 

 with a solution of dextrose of such strength that the final mixture, of 

 a uniform paste consistency, contained 0.2 per cent of dextrose. 

 The whole was then sterilized under 15 pounds pressure. Portions 

 of this paste were inoculated with 24-hour-old dextrose agar anaerobic 

 cultures of the bacteria, and covered with an inch layer of sterilized 

 fish broth made firm with 1.5 per cent agar and no nutriment, and 

 then incubated at 37^° C. until removed for analysis. Whenever a 

 sample was removed for analysis, the presence of the bacterium 

 with which it had been inoculated was determined positively. 



Table 8. — Ammonia and amines in pure cultures of Bacillus Walfischrauschbrand and 

 Bacillus B grown in the laboratory . 





Period 

 of incu- 

 bation. 



Total volatile alkaline material as— 



Nitro- 

 gen by 



amino 

 acid 

 method 

 as nitro- 

 gen per 



gram. 1 





Bacteria and culture No. 



Nitroge 



n per 100 grams. 



Percentage 

 of total. 



Alka- 

 linity 



(0.05 

 N.acid 



per 

 gram). 





Total. 



Am- 

 monia. 



Am- 

 ines. 



Am- 

 monia. 



Am- 

 ines. 



Sterile: 



30 



Days. 

 7 



2 

 4 

 7 

 7 



2 

 4 

 3 

 2 

 2 

 3 



Mg. 

 18.5 



259.5 



Mg. 



Mg. 



Per ct. 



Per ct. 



Mg. 

 0.55 



5.34 

 5.65 

 9.95 

 5.16 



Cc. 

 0.0 



B. Walfischrauschbrand: 



28 



215.2 



44.3 



82.9 



17.1 





28 





31 



510.1 

 334.6 



146.8 

 208.4 

 159.1 

 145.8 

 225.2 



447.1 

 286.2 



63.0 



48.4 



87.7 

 85.6 



12.3 

 14.4 



2.1 



32 



1.5 



Bacillus B : 



24 



1.2 



24 



160.3 

 127.7 

 132.6 

 198.8 



48.1 

 31.4 

 13.2 



26.4 



76.9 

 80.3 

 81.0 

 88.2 



23.1 

 19.7 

 9.0 

 11.8 





1.9 



25 





29 



2.52 

 5.07 

 5.59 





27 2 





27 



















1 These determinations were made by the latest amino acid method and apparatus. Van Slyke (J. 

 Biol. Chem. (1913) 16:121). 

 '' A few micrococci in this culture. 



