Desulfovibrio bastinii Magot et al.
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2. Perform all steps under anaerobic conditions.
3. Aseptically transfer 0.5 ml of #1250 broth to the vial and rehydrate the pellet. Transfer the suspension back into the broth tube. Inoculate a plate of a non-selective medium such as Tryptic Soy, Nutrient, or blood agar with 0.1 ml of the cell suspension.
4. Seal the tube with a rubber stopper and incubate anaerobically at 37oC. Incubate the plate(s) aerobically as a purity check.
5. After one or two days, growth should be evident as indicated by turbidity through out the broth. Once growth has been established the culture should be transferred to fresh broth every 24 to 48 hours.
6. This culture is very sensitive to oxygen when initially rehydrated, therefore steps should be taken to avoid exposure to oxygen. When the culture exhibits good growth it will remain viable for up to 1 week if stored at 4oC under anaerobic condition.
· 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.
· 100% nitrogen or 80% nitrogen-20% carbon dioxide gas mixture is typically employed as the oxygen free gas source.
When examined microscopically, the cells appear as typical single (some pairs) comma-shaped rods that are motile.
Always use freshly prepared anaerobic medium. If there is any question about the anaerobic condition of the medium, the medium can be reduced with the addition of 1.5% cysteine
(2.0 ml per 100 ml of medium).
Other commonly used reducing agents are sodium sulfide, cysteine, dithiothreitol, and titanium citrate. Cysteine is the reducing agent of choice since it does not cause the ferrous ammonium sulfate to precipitate.
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water sample collected in oil pipeline, Emeraude offshore oilfield, Congo
Michel Magot, personal communication
Magot M, et al. Desulfovibrio bastinii sp. nov. and Desulfovibrio gracilis sp. nov., moderately halophilic, sulfate-reducing bacteria isolated from deep subsurface oilfield water. Int. J. Syst. Evol. Microbiol. 54(Pt 5): 1693-1697, 2004. PubMed: 15388730