Clostridium cellulovorans Sleat et al.
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2. Under anaerobic conditions, withdraw approximately 0.5 to 1.0 ml of #1345 broth (5 to 6 ml) with a Pasteur or 1.0 ml pipette. Rehydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a #1345 agar slant and/or pre-reduced plate. A pre-reduced blood plate may also be inoculated with 0.1 ml of the cell suspension. An aerobic blood plate may also be streaked to check for purity.
5. Incubate the tubes and plates under anaerobic conditions at 37°C for 2 to 5 days.
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 connected 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.
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. A pink color observed in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred. Medium should be discarded.
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Sleat R, et al. Isolation and characterization of an anaerobic, cellulolytic bacterium, Clostridium cellulovorans sp. nov. Appl. Environ. Microbiol. 48: 88-93, 1984.
Validation list no. 17. Int. J. Syst. Bacteriol. 35: 223-225, 1985.
Tamaru Y, et al. Genome sequence of the cellulosome-producing mesophilic organism Clostridium cellulovorans 743B. J. Bacteriol. 192(3): 901-902, 2010. PubMed: 19948806