Clostridium phytofermentans Warnick et al.

1. Open vial.
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. A slant may be inoculated with 0.1 ml. Streak two agar plates to check for colonial morphology and purity.
4. Incubate tubes under an anaerobic atmosphere at 30^oC for 48 to 72 hours. Incubate one agar plate anaerobically for colony formation, and the other agar plate aerobically for a contamination check.
5. No growth should occur on the agar plate incubated aerobically.

ANAEROBIC CONDITIONS:

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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