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Culturing and Maintaining Clostridium difficile in an Anaerobic Environment
Published on: September 14, 2013
Continuous monitoring of oxygen concentrations in several systems for cultivation of anaerobic bacteria
Abstract:
Anaerobic chambers and jars are the two conventional methods used in clinical microbiology laboratories to produce an anaerobic atmosphere. The evacuation-replacement method, the Oxoid AnaeroGen, the Merck Anaerocult A, the BBL GasPak, the BBL GasPakPlus, the Adams Scientific GasGendicator, the Difco Anaerobic, and the bioMérieux Generbox anaer systems were compared for the timed decrease in the oxygen concentration in an anaerobic jar. The experiment was repeated 10 times for each system. The oxygen concentration was measured with an oxygen analyzer series 3600 instrument (Orbisphere Laboratories, Neuchâtel-Geneva Switzerland). The BBL GasPak, the BBL GasPakPlus, the bioMérieux Generbox, the Adams Scientific GasGendicator, and the Difco Anaerobic systems contain sodium borohydride, which liberates hydrogen. The Anaerocult A system contains iron powder which binds the oxygen. The active ingredient of the AnaeroGen system is ascorbic acid. The times to reach an O2 concentration of 0.5% were 8 to 15 min for the evacuation-replacement method, 26 to 41 min for the AnaeroGen system, 60 to 93 min for the Anaerocult A system, and 22 to 419 min for the sodium borohydride systems. The AnaeroGen system, the Anaerocult A system, and the evacuation-replacement method never failed to produce an anaerobic atmosphere. The sodium borohydride systems failed in 10 of 70 runs. These results suggest that the evacuation-replacement method or the Oxoid AnaeroGen system should be used to produce an anaerobic atmosphere. The Anaerocult A system showed a good reproducibility, but the length of time required to reach an appropriately low level of oxygen was too long. Because of the high failure rate, the borohydride systems cannot be recommended.
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