Propionibacterium freudenreichii subsp. shermanii (van Niel) Holdeman and Moore
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2. Under anaerobic conditions, withdraw 0.5 ml of recommended broth from a single test tube (5 to 6 ml) and rehydrate the entire vial contents.
3. Aseptically transfer this aliquot back into the broth tube. A slant and a pre-reduced blood plate may also be inoculated with 0.1 ml each of the cell suspension. An aerobic blood plate may also be streaked to check for purity.
4. Incubate tubes and one plate under anaerobic conditions at 37°C. Incubate one blood plate aerobically at 37°C.
5. Within 48 to 72 hours, growth is evident by a slightly turbid broth with smooth sediment. Colonies on the anaerobic blood agar plate are pinpoint, circular, entire, glistening, and transparent. No growth occurs on the blood agar plates incubated aerobically.
Anaerobic conditions for transfer may be obtained by either of the following:
· Use of an anaerobic gas chamber, or
· Placement of test tubes under a gassing cannula system hooked to anaerobic gas.
Anaerobic conditions for incubation may be obtained by any of the following:
· Loose screw caps on test tubes in anaerobic chamber,
· Loose screw caps on test tubes in an activated anaerobic gas pack jar, or
· Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
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Johnson JL, Cummins CS. Cell wall composition and deoxyribonucleic acid similarities among the anaerobic coryneforms, classical propionibacteria, and strains of Arachnia propionica. J. Bacteriol. 109: 1047-1066, 1972. PubMed: 5062339