Desulfarculus baarsii (Widdel) Kuever et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. Perform all steps under anaerobic conditions.
3. Aseptically transfer 0.5 ml of #1283 broth to the vial and rehydrate the pellet. Transfer the suspension back into the broth tube (5 to 6 ml). Inoculate a plate of a non-selective medium such as Trypticase Soy, Nutrient, Brain Heart Infusion or Blood agar with 0.1 to 0.2 ml.
4. Seal the test tube with a rubber stopper and incubate anaerobically at 37oC. Incubate the plate aerobically for a purity check.
5. After one to two days, growth is evident by turbidity through out the broth. When examined microscopically, the cells appear as single (some pairs) comma-shaped rods that are very motile. Once growth has been detected, the culture should be transferred to fresh broth(s). Growth should be detected within 24-48 hours.
6. This culture is very sensitive to oxygen when initially rehydrated, therefore steps should be taken to avoid exposure to oxygen. Once growth has been established, the culture should be transferred to fresh broth medium, every 48 hours. The culture will remain viable for up to 1 week at 4oC under anaerobic conditions, as long as the culture has established good growth before it is refrigerated.
· 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.
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Validation list no. 7. Int. J. Syst. Bacteriol. 31: 382-383, 1981.
Widdel FAnaerober abbau von fettsäuren und benzoesäure durch neu isolierte arten sulfat-reduzier-ender bakterien Dissertation, Gottingen, 1980