VIBRION SEPTIQUE 537 
point of injection, repeated if necessary, is usually the most direct 
method of obtaining cultures unless the strain has lost its pathogenicity 
through prolonged cultivation outside the body. It is also possible to 
allow mixed cultures to sporulate in media containing tissue or cooked 
meat; heat to 80° C, for thirty minutes, and inoculate the heated sus- 
pension upon agar-blood or agar-ascitic fluid plates. The well isolated 
colonies should be purified finally by the method of development from 
a single cell, to be certain that an admixture with B. sporogenes or 
other anaerobic symbiote is prevented. 
The organism tends to form a delicate, spreading film upon the sur- 
face of solid media, hence pure cultivations from surface colonies are 
difficult to obtain. There is nothing particularly characteristic about 
surface or submerged colonies in solid media. Deep colonies, however, 
tend to become filamentous in media that are not too rigid. Gelatin 
is softened by most strains, to the point where it will no longer solidify, 
but there is no evidence that a soluble gelatinase forms. The decom- 
position of protein, as measured by the increase in amino-acid forma- 
tion and free ammonia is very slight. The acidity of the organism in 
liberating amino-acids is distinctly less than that of B. welchii.^ 
]\rilk is rather slowly fermented; there is usually a slight evolution of 
gas (principally CO2 and Ho) , which is very much less rapid than that 
of B. welchii. The casein is coagulated and precipitated. The reac- 
tion of the medium becomes progressively acid. An odor of butyric 
acid develops. 
Both acid and some gas are produced from the hexoses— glucose, 
fructose and galactose. Maltose and lactose of the biose series are 
also fermented; salicin is also utilizable as a non-nitrogenous source 
of energy. Glycerin, the alcohols of the hexose series (mannitol, 
dulcitol and sorbitol), and starch, dextrin and inulin appear to be 
untouched. With reference to saccharose there appears to be differ- 
ence of opinion. Miss Robertson^ states that gas is not produced in 
this sugar, and this observation is in accord with that of Achalme,^ Jun- 
gano and Distaso.* and Hall.^ 
Products of Growth.— Vibrion septique is not a proteolytic organism. 
There is no evidence that proteolytic enzymes are produced, and chemi- 
cal evidences of the action of the vibrion upon the nitrogenous consti- 
tuents of media, even in the absence of utilizable carbohydrate, are 
minimal." The gas produced from carbohydrates is principally H2 
and CO2. 
In media containing cooked meat some gas is formed, and the meat 
becomes tinged with a bright pink coloration. The meat is not black- 
ened and a distinct odor of butyric acid is noticeable. The pink colora- 
tion, however, is not usually noticed in the milk coagulum; this may 
be a pohit of distinction between it and the Welch bacillus. 
1 Kendall, Day and Walker: Jour. Inf. DJs., 1922, 30, No. 2. 
2 British Med. Jour., 1918, i, 583. » Ann. Inst. Pasteur., 1902, 16, 633. 
* Les Anaerobies, Paris, 1910, p. 78. ^ jour. Infec. Dis., 1922, 30, 445. 
6 Wolf: Jour. Pathol, and Bacteriol., 1918, 22, 115. 
