Propionibacterium freudenreichii subsp. shermanii (van Niel) Holdeman and Moore

1. Open thawed vial according to enclosed instructions or visit www.atcc.org for instructions.
2. Under anaerobic conditions 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 brucella blood plate and/or brucella blood agar slant.
4. Incubate in an anaerobic atmosphere at 30°C for 24 to 48 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

Anaerobe Systems Brucella Blood Plate (AS-111 or AS-141) is highly recommended to analyze colony morphology and purity. This strain does not grow well on ATCC Medium #260: Tryptic Soy Agar/Broth with 5% Sheep Blood (defibrinated).

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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