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 WALFISCHRAUSCHBRAND AND 
Baciiuus 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 Walfischrauschbrand 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 374° 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 1n the laboratory. 
| 
Total volatile alkaline material as— 
| Nitro- 
gen by | Alka- 
amino | linity 
Nitrogen per 100 grams. a ar acid | (0.05 
; method| N. acid 
aSnitro-| per 
gen per} gram). 
gram, 
Period 
Bacteria and culture No. of incu- 
bation. 
Am- Am- Am- | Am- 
Total. | monia.| ines. | monia.| ines. 
Sterile: Days. Mg. Mg. Mg. | Perct.| Perct.} Mog. Ce. 
7 le 0. 55 I: 
feng oe oe ee OU eP COE ee Sete SEES ne 2 || 1462 82) S252 2. Sa | Se ee tos eae eet 1.2 
Barre LYONS et BE 4] 208.4] 160.3| 481| 769) 23.1 ]..2....- 1.9 
UAE sec ccc coos Secet es Scece sSecsse 3 159gb 27 31.4 80. 3 LOR iee = oe =a) ae sos oe 
DOSES See oe ao cele letae area lee 2{ 145.8} 132.6 13. 2 81.0 9.0 Lai ESR R Te 
Zhe ee eR so ee ee oats « eed 2| 225.2} 198.8 26. 4 88. 2 11.8 AU Sees 
DCMS SL SY ES | Bite ora Ss (ad. speed: faceeme: 5.59 |-. ssa. 
: I } 
a 1These 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. 
