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Optimisation of culture methods for the selective isolation of diarrhoeagenic Escherichia coli from water

Diarrhoea caused by strains of Escherichia coli is recognised as a significant cause of morbidity and mortality world-wide. Efficient isolation of these bacteria from water is necessary for effective management of available water sources and for the control and prevention of waterborne disease. Methods that are currently available do not allow for the efficient isolation of the diarrhoeagenic E. coli. Therefore, the aims of the current study were: to determine the efficacy of Chromocult® Coliformen Agar for identification of diarrhoeagenic E. coli; correlate virulence gene carriage with biochemical profiles of diarrhoeagenic E. coli; and develop an enrichment method for the selective isolation of diarrhoeagenic E. coli. Chromocult® Coliformen Agar proved unreliable for identifying diarrhoeagenic E. coli – 30% of strains would have been incorrectly identified using this medium. The carriage of pathotype-specific virulence genes by enteroaggregative E. coli strains was associated with the production of ornithine decarboxylase and the fermentation of sucrose. The enrichment procedure detailed in this study was an improvement upon an existing method. In addition, the improved method proved effective for the isolation of five diarrhoeagenic E. coli pathotypes. In contrast, the original procedure was only shown to be effective when isolating enterohaemorrhagic E. coli. The improved method was applied on environmental water and two samples yielded colonies positive for genes carried by enteropathogenic E. coli and enterotoxigenic E. coli, respectively. As confirmation of these results PCR products were sequenced and corresponded to published sequences of the virulence genes. / Ms. P.S. Kay Dr. T. G. Barnard Prof. P. Jagals

Identiferoai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:uj/uj:7661
Date13 August 2008
CreatorsHeine, Lee
Source SetsSouth African National ETD Portal
Detected LanguageEnglish
TypeThesis

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