Bacteroides fragilis (Veillon and Zuber) Castellani and Chalmers

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 #1490 broth. Aseptically transfer the entire contents to a 5-6 mL tube of #1490 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 24 hours. Incubate one agar plate aerobically at 37°C to check for contamination.

• 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

Always use freshly prepared pre-reduced media or pre-reduced media that has been previously prepared but stored under anaerobic conditions.  Resazurin in the media is a color indicator for anaerobic conditions.  Observance of pink color in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred.  Medium should be discarded.

Additional information on this culture is available on the ATCC^® web site at www.atcc.org.

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