Campylobacter concisus Tanner et al.

1. Add formate/fumarate supplement to all media to be used, as described above.
2. Open vial.
3. Under anaerobic conditions, withdraw 0.5 mL from a single tube of #1540 broth and rehydrate the entire pellet.
4. Aseptically transfer this aliquot back into the broth tube. Additional broth can be inoculated using 0.2 mL of this cell suspension per tube. A slant and a pre-reduced blood plate may also be inoculated with 0.1 mL each of the cell suspension. An aerobic blood plate may also be streaked to check for purity.
5. Incubate the broth tubes, slants, and plates at 37°C under anaerobic conditions. You may use an anaerobe jar with catalyst and gas generator pack or other suitable means of producing anaerobic conditions. Be sure caps are loosened to facilitate gas exchange. Incubate aerobic blood plate aerobically at 37°C.
6. Initially, 3 to 7 days of incubation are required before visible is evident. A slightly turbidity is seen in the broth and growth is observed on the agar surface. No growth should be seen on the aerobic plate.

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.
• 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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