Zymomonas mobilis subsp. mobilis (Lindner) Kluyver and van Niel

1. Open vial according to enclosed instructions.
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 of broth may be inoculated with 0.5 mL each from the suspension, and 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 30°C.  Incubate one agar plate anaerobically for colony formation, and one aerobically as a purity check.
5. Within 24-48 hours, growth should be evident by turbidity in the broth tube with slight gas formation.  No growth should occur on agar plates 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,
• Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.

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