"Desulfovibrio magneticus" Sakaguchi et al.
700980 ™
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700980 ™
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2. Perform all steps under anaerobic conditions.
3. Aseptically transfer 0.5 ml of #2225 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 Trypticase 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 30oC. Incubate the plate(s) aerobically as a purity check.
5. After six to eight weeks, growth should be evident as indicated by turbidity that settles to the bottom of the tube and is easily resuspended when the tube is inverted. Once growth has been established the culture should be transferred to fresh broth every 4 to 6 weeks.
6. This culture is very sensitive to oxygen when initially rehydrated; therefore steps should be taken to avoid exposure to oxygen.
ANAEROBIC CONDITIONS:
· 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-10% carbon dioxide-10% hydrogen gas mixture is typically employed as the oxygen free gas source.
When examined microscopically, the cells appear as singles, 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 0.1 ml of a 3% stock solution of cysteine per 5-6 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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Sakaguchi T, et al. A novel method for isolation of magnetic bacteria without magnetic collection using magnotaxis. J. Microbiol. Methods 26: 139-145, 1996.
Sakaguchi T, et al. Magnetite formation by a sulphate-reducing bacterium. Nature 365: 47-49, 1993.
Kawaguchi R, et al. Phylogenetic analysis of sulfate-reducing magnetic bacterium, RS-1, demonstrates its membership of the delta-Proteobacteria. FEMS Microbiol. Lett. 126: 277-282, 1995.
Sakaguchi T, et al. Desulfovibrio magneticus sp. nov., a novel sulfate-reducing bacterium that produces intracellular single-domain-sized magnetite particles. Int. J. Syst. Evol. Microbiol. 52: 215-221, 2002. PubMed: 11837306
type strain