Bifidobacterium boum Scardovi et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of the recommended broth from a single test tube (5 to 6 ml) and rehydrate the vial contents.
3. Aseptically transfer this aliquot back into the broth. Additional tubes may be inoculated with 0.5 ml each from the suspension. 0.1 ml may also be inoculated onto a slant. Streak several blood plates to check for colonial morphology and purity.
4. Incubate tubes under an anaerobic atmosphere at 37oC. Incubate one agar plate anaerobically for colony formation, and one aerobically for aerobic contamination check.
5. In 24 to 48 hours, growth should be evident by good turbidity in the broth and by colonies on the anaerobic agar surfaces. After 48 hours, the colonies will be white, glistening, with irregular edges. No growth should occur on the aerobic plate.
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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Scardovi V, et al. Bifidobacterium cuniculi, Bifidobacterium choerinum, Bifidobacterium boum, and Bifidobacterium pseudocatenulatum: Four new species and their deoxyribonucleic acid homology relationships. Int. J. Syst. Bacteriol. 29: 291-311, 1979.
Skerman VB, et al. Approved lists of bacterial names. Int J Syst Bacteriol 30: 225-420, 1980.