Methanocaldococcus vulcanius (Jeanthon et al.) Whitman
700851 ™
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700851 ™
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2. If needed exchange the gas in the test tube for 80% H2-20% CO2.
3. If the medium is pink (see discussion about resazurin) add 2.0 ml of reducing agent (3% cysteine, stock solution) per 100 ml of medium. Let the medium sit at room temperature for 10 to 20 minutes, until the resazurin becomes colorless, before inoculating.
4. Open the frozen vial and immediately place the vial under a stream of sterile gas, 80% H2-20% CO2 to maintain anaerobicity.
6. Growth should be detected in the broth within 48 hours. No growth should be detected on the aerobic plate or broth.
ANAEROBIC CONDITIONS:
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.
1. Displace the dead space in the syringe with a sterile
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Jeanthon C, et al. Methanococcus vulcanius sp. nov., a novel hyperthermophilic methanogen isolated from East Pacific Rise, and identification of Methanococcus sp. DSM 4213T as Methanococcus fervens sp. nov.. Int. J. Syst. Bacteriol. 49: 583-589, 1999. PubMed: 10319479
type strain
. Validation of publication of new names and new combinations previously effectively published outside the IJSEM.. Int. J. Syst. Evol. Microbiol. 52: 685-690, 2002. PubMed: 12054225
Whitman WBGenus I. Methanocaldococcus gen. nov.In: Whitman WBBergey's manual of systematic bacteriology2nd ed.New YorkSpringer-Verlag243-245, 2001
DSMZ, personal communication