Clostridium haemolyticum (Hall) Scott 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 entire vial contents.
3. Aseptically transfer this aliquot back into the broth. Additional tubes may be inoculated with 0.5 ml each from the suspension. Also, 0.1 ml may 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. Within 24 to 48 hours, growth should be evident by turbidity in the broth. No growth should occur on 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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