Desulfovibrio aespoeensis Motamedi and Pedersen
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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. (see below)
3. Aseptically transfer 0.5 ml of ATCC Medium #2108 to the vial and rehydrate the freeze-dried pellet. Transfer the suspension back into the tube of broth. Inoculate a plate of non-selective medium with 0.1 of the culture.
4. Seal the test tube with a rubber stopper and incubate anaerobically at 30oC. Incubate the plate(s) aerobically as a purity check.
5. Growth could be detected within 24 hours. Cells appear as vibriod shaped rods in singles and pairs. The cells are motile. Good growth is not obtained on agar.. Once growth has been established, the culture should be transferred to fresh broth every 24 to 48 hours.
· 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.
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