Fibrobacter succinogenes (Hungate) Montgomery 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. Under anaerobic conditions, withdraw 0.5 ml of the recommended broth from a single test tube (5 to 6 ml) and rehydrate the entire vial contents.
3. Aseptically transfer this aliquot back into the broth. Additional tubes may be inoculated with 0.5 ml each from the suspension. Also, 0.1 ml may be inoculated onto a slant. Streak one blood plate to check for any aerobic contaminants.
4. Incubate tubes under an anaerobic atmosphere at 37oC. Incubate one agar plate aerobically for aerobic contamination check.
Anaerobic conditions for transfer may be obtained by either of the following:
· Use of an anaerobic gas chamber, or
· Placement of test tubes under a gassing cannula system hooked to anaerobic gas.
Anaerobic conditions for incubation may be obtained by any of the following:
· Loose screw caps on test tubes in anaerobic chamber,
· Loose screw caps on test tubes in an activated anaerobic gas pack jar, or
· Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
We suggest using Co-enyme M and adding that reducing agent to the medium at least one hour before the medium is to be inoculated. Co-enzyme M (mercaptoethanesulfonic acid) (100 X solution): Dissolve 5.0 g in 100 ml of deionized water. Distribute into screw cap test tubes, 56 ml per tube and seal with rubber stoppers under N2 gas. Autoclave to sterilize. Excess tubes can be stored at room temperature for up to 2 months.
Additional information on this culture is available on the ATCC web site at www.atcc.org.
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Amann RI, et al. Diversity among Fibrobacter isolates: Towards a phylogenetic classification. Syst. Appl. Microbiol. 15: 23-31, 1992.