Clostridium tyrobutyricum van Beynum and Pette

1. Open vial according to enclosed instructions or visit www.atcc.org for instructions.
2. Under anaerobic conditions aseptically rehydrate the entire pellet with approximately 0.5 mL of #2107 broth. Aseptically transfer the entire contents to a 5-6 mL tube of #2107 broth. Additional test tubes can be inoculated by transferring 0.5 mL of the primary broth tube to these secondary broth tubes. Best practice dictates the use of pre-reduced media.
3. Use several drops of the primary broth tube to inoculate a #260 plate and/or #260 agar slant.
4. Incubate in an anaerobic atmosphere at 37°C for 48 hours. Incubate one agar plate aerobically at 37°C to check for contamination.

Anaerobic conditions for incubation may be obtained by any of the following:

• Loose screw caps on test tubes in an anaerobic chamber
• Loose screw caps on test tubes in an activated anaerobic gas pack jar
• Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained

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