Acetobacterium fimetarium Kotsyurbenko et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of the recommended broth from a single tube (5 to 6 ml) and rehydrate the vial contents.
4. Incubate tubes under an anaerobic atmosphere at 20oC. Incubate one agar plate anaerobically for colony formation, and one aerobically for aerobic contamination check.
5. In 48 hours, growth should be evident by turbidity that settles to the bottom of the tube. No growth should occur on agar plate incubated aerobically.
a. Balch tubes (available from Bellco Glass, Vineland, NJ; are specially designed for anaerobic work and use an aluminum crimp cap to hold a rubber stopper in place. Needles can easily be inserted through the stopper, and the tubes can be pressurized to 2 atm. Alternatively, serum vials may be used, or screw cap tubes with butyl rubber stoppers, in the latter case the stopper may be removed and the tube placed under a cannula system that dispenses sterile, oxygen free gas for addition of reducing agents or inoculation.
b. Resazurin is a commonly used redox indicator that is pink when the redox potential is above 50 mv, and colorless when the redox potential is below 110 mv, i.e. highly reducing. Most strict anaerobes require this low redox potential for optimum growth.
c. 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.
d. Syringes can be made anaerobic by one of two methods.
1. Displace the dead space in the syringe with a sterile
2. Displace the dead space in the syringe with a
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Validation list no. 60. Int. J. Syst. Bacteriol. 47: 242, 1997.
Kotsyurbenko OR, et al. New species of psychrophilic acetogens: Acetobacterium bakii sp. nov., A. paludosum sp. nov., A. fimetarium sp. nov.. Arch. Microbiol. 163: 29-34, 1995.