Clostridium proteolyticum Jain and Zeikus
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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 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 plate under anaerobic conditions at 37°C. Incubate blood plate aerobically at 37°C.
5. In 24 to 48 hours, growth is evident by turbidity in the broth. On agar surfaces, colonies are convex, circular, shiny, entire, clear, and smooth. No growth should occur on the blood agar plate 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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Validation list no. 26. Int. J. Syst. Bacteriol. 38: 328-329, 1988.
Jain MK, Zeikus JG. Taxonomic distinction of two new protein specific, hydrolytic anaerobes: Isolation and characterization of Clostridium proteolyticum sp.nov. and Clostridium collagenovorans sp.nov.. Syst. Appl. Microbiol. 10: 134-141, 1988.