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Functional cloning of the dihydropteroate synthase gene of Staphylococcus haemolyticus
Authors:Paul Kellam  Walter S Dallas  Stuart P Ballantine  Chris J Delves
Institution:Department of Genetics and Microbiology, Life Science Building, University of Liverpool, P.O. Box 147, Liverpool L69 3BX, UK; Institute of Freshwater Ecology, Windermere Laboratories, Cumbria, UK
Abstract:Abstract The usefulness of oxonol ( bis -(1,3-dibutylbarbituric acid)trimethine oxonol) as a generally applicable indicator of bacterial viability was investigated using untreated and killed cultures of a variety of bacterial genera. Killing methods involved either heat or bactericidal antibiotics. For all strains tested, the fluorescent dye showed significantly more intense staining of killed than untreated cells. The sensitivity of Aeromonas salmonicida to gentamicin was assessed using oxonol. Although the bacterium was shown to be sensitive to the antibiotic, there was a delay between the time cells lost culturability, as judged by numbers of colony forming units, and that for which a dead cell population could be detected by flow cytometry.
Keywords:Bacterial viability  Oxonol dye  Flow cytometry  Membrane potential
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