Development and application of a spiral plating method for the enumeration of Escherichia coli O157 in bovine faeces

Aim: To develop and validate a direct plating method applicable to epidemiological studies for enumerating Escherichia colt O157 in cattle faeces. Methods and Results: The spiral plate count method was used to enumerate E. colt O157 in faecal samples. The accuracy and variation of counts was then assessed using faecal samples inoculated with E. colt O157. There was good agreement between inoculated levels of E. colt O157 and those recovered from faeces, particularly when counts were >10(2) CFU g(-1) of faeces. The method was applied to a small study assessing short-term survival of E. colt O157 in naturally infected cattle faeces. E. colt O157 was found to survive in faeces for over 10 days at concentrations above 10(3) CFU g(-1) of faeces. Populations of E. colt O157 were also found to increase 100-fold in the first few hours after defecation. Conclusions: The enumeration method is easy to implement and enables a quick throughput of large numbers of samples. The method is accurate and reliable and enables the inherent variation in count data to be explored but needs to be used in combination with a more sensitive method for samples containing <10(2) CFU g(-1) of faeces. Significance and Impact of the Study: The method described is appropriate for enumeration of E. colt O157 in cattle faeces in large-scale epidemiological studies.