Sedimentibacter hydroxybenzoicus (Zhang et al.) Breitenstein et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of 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 tube. A slant and a pre-reduced blood plate may also be inoculated with 0.1 ml each of the cell suspension. An aerobic blood plate may also be streaked to check for purity.
4. Incubate tubes and plate under anaerobic conditions at 37°C. Incubate blood plate aerobically at 37°C.
5. Within 24 to 48 hours, growth should be evident by turbidity in the broth. No growth should occur on the blood agar plate incubated aerobically.
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.
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Zhang X, Wiegel J. Isolation and partial characterization of a Clostridium species transforming a para-hydroxybenzoate and 3,4-dihydroxybenzoate and producing phenols as the final transformation products. Microb. Ecol. 20: 103-121, 1990.
freshwater lake near Athens
Zhang X, et al. Clostridium hydroxybenzoicum sp. nov., an amino acid-utilizing, hydroxybenzoate-decarboxylating bacterium isolated from methanogenic freshwater pond sediment [published erratum appears in Int. J. Syst. Bacteriol. 44: 854, 1994]. Int. J. Syst. Bacteriol. 44: 214-222, 1994. PubMed: 8186087
Zhang X, Wiegel J. Reversible conversion of 4-hydroxybenzoate and phenol by Clostridium hydroxybenzoicum. Appl. Environ. Microbiol. 60: 4182-4185, 1994.
Huang J, He Z, Wiegel J. Cloning, characterization, and expression of a novel gene encoding a reversible 4-hydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum. J. Bacteriol. 181(16): 5119-5122, 1999. PubMed: 10438791