Desulfotomaculum thermobenzoicum subsp. thermobenzoicum Tasaki et al.
49756 ™
49756 ™
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2. If needed exchange the gas in the test tube for 80% N2-20% CO2.
3. If the medium is pink (see discussion about resazurin) add 0.1 ml of reducing agent (3% cysteine, stock solution) per each 5-6 ml of medium. Let the medium sit at room temperature for 20 to 30 minutes - until the resazurin becomes colorless - before inoculating.
4. When the Balch tube is ready to inoculate, open the vial according to enclosed instructions.
5. For inoculation, use a 1.0 ml syringe tipped with 22 gauge needle. Make the syringe anaerobic (see discussion below) and withdraw 0.5 ml of #1282 broth and use this to rehydrate the freeze-dried pellet using anaerobic techniques. Transfer the rehydrated cell suspension back to a tube of broth and incubate at 60oC. Plate 0.1 ml of the inoculated culture onto a non-selective medium and incubate aerobically at 37oC. Inoculate a nonselective anaerobic and aerobic broth and incubate at 60oC
6. Growth should be detected in the broth within 48 to 72 hours. There should be no growth detected on the aerobic plate. There should be no growth in the nonselective aerobic or anaerobic broth.
ANAEROBIC CONDITIONS:
a. Balch tube refers to a special type of test tube that is designed to be pressurized and is suited for anaerobic work. The Balch test tubes can be purchased form Bellco glass (www.bellcoglass.com; stock no. 2048-00150).
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.
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
Cells are rods in singles and pairs.
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Tasaki M, et al. Isolation and characterization of a thermophilic benzoate-degrading sulfate-reducing bacterium Desulfotomaculum thermobenzoicum new species. Arch. Microbiol. 155: 348-352, 1991.
Deed RW, et al. Validation list no. 39. Int. J. Syst. Bacteriol. 41: 580-581, 1991. PubMed: 1742202
Plugge CM, et al. Desulfotomaculum thermobenzoicum subsp. thermosyntrophicum subsp. nov., a thermophilic, syntrophic, propionate-oxidizing, spore-forming bacterium. Int. J. Syst. Evol. Microbiol. 52: 391-399, 2002. PubMed: 11931147
type strain