Low-temperature-growing clostridia from marine sediments

Abstract

Marine sediments inoculated into cooked meat medium were enriched for 5 months at 0–5 °C. Organisms isolated and purified were tested for growth at low temperatures, and those growing only anaerobically at 5 °C and below were retained for further study. The growth–temperature range was determined by growing 18 isolates on a temperature-gradient incubator over a temperature range of 0 to 34 °C. On the bases of minimum, optimum, and maximum growth temperatures, the 18 isolates were separated into obligate and facultative psychrophilic and psychrotrophic clostridia. The DNA was extracted from cells grown at 12 °C and the percentage of G + C for all the isolates fell within the range characteristic of Clostridium species. The volatile alcohols and acids produced on a glucose medium were detected by gas chromatography. Ten of the isolates produced no alcohols and only acetic and butyric acids. One isolate produced butanol and acetic and butyric acids. The remaining seven isolates produced one or two alcohols, ethanol and butanol, and up to six acids. The 18 isolates fell into distinct groups with a close relationship between growth–temperature range and volatile alcoholic and acidic products. Biochemical tests showed that the isolates resembled Clostridium species described in the literature; however, they were not like any known species and were not named.