Treponema socranskii subsp. buccale Smibert et al.
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2. Under anaerobic conditions, aseptically withdraw approximately 0.5 to 1.0 ml from a tube of #1367 broth. Rehydrate the entire pellet.
3. Transfer this aliquot back into the broth tube.
4. Additional tubes of broth may be inoculated with 0.5 ml each of the suspension. Inoculate two Tryptic Soy Agar plates with 5% sheep blood (ATCC Medium #260). Incubate one blood plate anaerobically to check for colony formation and incubate the second plate aerobically to check for purity.
5. Incubate tubes and one plate under anaerobic conditions at 37°C. Incubate the second blood plate aerobically.
6. Growth occurs within 7 days, and should be evident by very slight turbidity in the broth and helical cells under phase contrast microscopy. The aerobic plate should show no signs of growth.
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, or
· Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
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Smibert RM, et al. Treponema socranskii sp. nov., Treponema socranskii subsp. socranskii subsp. nov., Treponema socranskii subsp. buccale subsp. nov., and Treponema socranskii subsp. paredis subsp. nov. isolated from the human periodontia. Int. J. Syst. Bacteriol. 34: 457-462, 1984.
Wyss C, et al. Treponema amylovorum sp. nov., a Saccharolytic spirochete of medium size isolated from an advanced human periodontal lesion. Int. J. Syst. Bacteriol. 47: 842-845, 1997.