CHAMBERLAIN, ROBERT E. (The National Drug Co., Swiftwater, Pa.). Evaluation of live tularemia vaccine prepared in a chemically defined medium. Appl. Microbiol. 13:232-235. 1965.-A chemically defined medium was prepared which adequately supported growth of a vaccine strain of Pasteurella tularensis. This medium differed from those previously described in: (i) concentration of components, (ii) a requirement for calcium pantothenate to obtain increased growth, and (iii) a low initial pH. Varying the concentration of individual components up to 10 times the standard amount did not increase the viable population or affect dissociation. The vaccine strain grown in this chemically defined medium, although lower in viable population, appears to retain its identity and to be equal in potency to that prepared by the conventional method. This preliminary study indicates the potential utility of this medium as a basis for controlled studies of a live bacterial vaccine in terms of growth characteristics, dissociation, virulence, and immunogenicity.
A chemically defined medium was prepared which adequately supported growth of a vaccine strain of
Pasteurella tularensis
. This medium differed from those previously described in: (i) concentration of components, (ii) a requirement for calcium pantothenate to obtain increased growth, and (iii) a low initial
p
H. Varying the concentration of individual components up to 10 times the standard amount did not increase the viable population or affect dissociation. The vaccine strain grown in this chemically defined medium, although lower in viable population, appears to retain its identity and to be equal in potency to that prepared by the conventional method. This preliminary study indicates the potential utility of this medium as a basis for controlled studies of a live bacterial vaccine in terms of growth characteristics, dissociation, virulence, and immunogenicity.
Results of minimal inhibitory concentration tests with a diversity of bacterial strains showed that 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390) inhibited the growth of all microorganisms tested (other than Pseudomonas aeruginosa) at 25 jug/ml or less, whereas MICs obtained for P. aeruginosa ranged from 50 to >100 ,ug/ml. Therefore, C-390 was evaluated as a potential selective agent for P. aeruginosa in pseudomonas agar F. Recovery tests were conducted on this medium with 53 strains of P. aeruginosa, and the results were compared to those obtained in similar tests on commercially available selective media, i.e., pseudomonas isolation agar and Pseudosel agar. The results of these comparisons indicated that pseudomonas agar F with C-390 was significantly less inhibitory than Pseudosel agar and pseudomonas isolation agar and more selective than pseudomonas isolation agar. The incorporation of C-390 in pseudomonas agar F also provided a medium that was both selective and differential. Preliminary evidence also suggested that C-390 may be added to other basal media with comparable results.
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