Brachyspira hyodysenteriae (Harris et al.) Ochiai et al.

1. Open thawed vial.
2. Under anaerobic conditions aseptically transfer the entire contents to a 5-6 mL tube of #1827 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 #260 plates and 0.5 mL of the suspension to #1827 slants, making a biphasic culture.
4. Incubate in an anaerobic atmosphere at 37°C for 2-5 days. Incubate one agar plate aerobically at 37°C to check for contamination.
5. Growth can be observed as light turbidity in the broth and as a thin film on the slants and colonies on the #260 plate. The aerobic plate should show no signs of growth.

ANAEROBIC CONDITIONS:

Anaerobic conditions for transfer may be obtained by the use of an anaerobic gas chamber or placement of test tubes under a gassing cannula system connected to anaerobic gas.

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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