Clostridium puniceum Lund et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of #1869 from a single test tube (5 to 6 ml) and rehydrate the vial contents.
3. Aseptically transfer this aliquot back into the broth tube. Additional tubes may be inoculated with 0.5 ml each from the suspension. A slant of #1869 may also be inoculated with 0.2 ml. Streak several blood plates to check for colonial morphology and purity.
4. Incubate tubes and one blood plate under an anaerobic atmosphere at 26oC. Incubate the other blood plate in air.
5. Within 48 hours, growth is evident by turbidity in the broth and by colonies on the anaerobic agar surfaces. On #260 agar, colonies are entire, raised, and slightly rough with a light pink hue. No growth should occur on the agar plate incubated in air.
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.
For best anaerobic growth, use freshly prepared #260 agar or store freshly prepared media under anaerobic conditions prior to use. Medium #1869 contains resazurin. Resazurin is a color indicator for anaerobic conditions. Observance of pink color in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred. Medium should be discarded.
Additional information on this culture is available on the ATCC web site at www.atcc.org.
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If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.
Lund BM, et al. Characterization of strains of Clostridium puniceum sp.nov., a pink-pigmented, pectolytic bacterium. J. Gen. Microbiol. 122: 17-26, 1981.
Validation list no. 6. Int. J. Syst. Bacteriol. 31: 215-218, 1981.
Young JM, et al. Names of plant pathogenic bacteria 1864-1995. Rev. Plant Pathol. 75: 721-763, 1996.