Slackia heliotrinireducens (Lanigan) Wade et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of recommended broth from a single test tube (5 to 6 ml) and rehydrate the vial contents.
3. Aseptically transfer this aliquot back into the broth tube. Additional tubes may be inoculated with 0.5 ml each from the suspension. A slant of #2008 may also be inoculated with 0.1 ml. Streak several blood plates to check for colonial morphology and purity.
4. Incubate tubes under an anaerobic atmosphere at 37oC. Incubate one agar plate anaerobically for colony formation, incubate the second aerobically for an aerobic contamination check.
5. Within 24 to 48 hours, growth should be evident by faint turbidity in the broth tube. Cells in broth appear as spherical to slightly elongated cocci, occurring singly, in pairs, in small clusters and occasionally in short chains of 3 to 6 cells. The anaerobic plate shows colonies that are pinpoint, entire, colorless , and transparent, with no hemolysis. No growth should occur on agar plates incubated aerobically.
· To obtain a fully reduced medium, it is necessary that the medium be anoxic and that a reducing agent be added. Common reducing agents are sodium sulfide, cysteine, dithiothreitol, and titanium citrate.
· Tubes of media are placed under a gassing cannula system hooked to a source of oxygen free gas.
· All transfers are performed while the test tubes are on the cannula system with a gentle stream of oxygen free gas flowing through the system.
· As the test tubes are removed from the cannula system each is sealed with butyl rubber stopper thus maintaining the anaerobic headspace.
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Lanigan GW. Peptococcus heliotrinreducans, sp. nov., a cytochrome-producing anaerobe which metabolizes pyrrolizidine alkaloids. J. Gen. Microbiol. 94: 1-10, 1976. PubMed: 945326
Validation list no. 10. Int. J. Syst. Bacteriol. 33: 438-440, 1983.
Ezaki T, Yabuuchi E. Transfer of Peptococcus heliotrinreducens corrig. to the genus Peptostreptococcus: Peptostreptococcus heliotrinreducens, Lanigan 1983 comb. nov. Int. J. Syst. Bacteriol. 36: 107-108, 1986.
Wade WG, et al. The family Coriobacteriaceae: reclassification of Eubacterium exiguum (Poco et al. 1996) and Peptostreptococcus heliotrinreducens (Lanigan 1976) as Slackia exigua gen. nov., comb. nov. and Slackia heliotrinireducens gen. nov., comb. nov., and Eubacteriumlentum (Prevot 1938) as Eggerthella lenta gen. nov., comb. nov.. Int. J. Syst. Bacteriol. 49: 595-600, 1999. PubMed: 10319481