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  • Title: Production of types A and B spores of Clostridium botulinum by the biphasic method: effect on spore population, radiation resistance, and toxigenicity.
    Author: Anellis A, Berkowitz D, Kemper D, Rowley DB.
    Journal: Appl Microbiol; 1972 Apr; 23(4):734-9. PubMed ID: 4111814.
    Abstract:
    Spores of three strains each of type A and type B Clostridium botulinum were produced both by a biphasic (solid medium overlaid with an aqueous phase) and by a "conventional" (deep broth culture) procedure. Sporogenesis by the biphasic system was more rapid, convenient, and economical, and yielded as many or more heat-resistant (80 C, 10 min) spores per milliliter as by the conventional technique. Of several aqueous phases [thiamine-hydrochloride, yeast extract, (NH(4))(2)SO(4)] tested with strain 62A, the highest spore colony counts were obtained with 2.0% (NH(4))(2)SO(4). The six strains formed maximum spore numbers in 5 to 6 days of incubation. Spores produced by the two methods had essentially equal radiation resistances (D and lag values), and their subcultures gave similar toxin titers (LD(50) values).
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