Escherichia coli isolates (n ؍ 658) obtained from drinking water intakes of Comox Lake (2011 to 2013) were screened for the following virulence genes (VGs): stx 1 and stx 2 (Shiga toxin-producing E.
coli [STEC]), eae and the adherence factor (EAF) gene (enteropathogenic E. coli [EPEC]), heat-stable (ST) enterotoxin (variants STh and STp) and heat-labile enterotoxin (LT) genes (enterotoxigenic E. coli [ETEC]), and ipaH (enteroinvasive E. coli [EIEC]). The only genes detected were eae and stx 2 , which were carried by 37.69% (n ؍ 248) of the isolates. Only eae was harbored by 26.74% (n ؍ 176) of the isolates, representing potential atypical EPEC strains, while only stx 2 was detected in 10.33% (n ؍ 68) of the isolates, indicating potential STEC strains. Moreover, four isolates were positive for both the stx 2 and eae genes, representing potential EHEC strains. The prevalence of VGs (eae or stx 2 ) was significantly (P < 0.0001) higher in the fall season, and multiple genes (eae plus stx 2 ) were detected only in fall. Repetitive element palindromic PCR (rep-PCR) fingerprint analysis of 658 E. coli isolates identified 335 unique fingerprints, with an overall Shannon diversity (H=) index of 3.653. Diversity varied among seasons over the years, with relatively higher diversity during fall. Multivariate analysis of variance (MANOVA) revealed that the majority of the fingerprints showed a tendency to cluster according to year, season, and month. Taken together, the results indicated that the diversity and population structure of E. coli fluctuate on a temporal scale, reflecting the presence of diverse host sources and their behavior over time in the watershed. Furthermore, the occurrence of potentially pathogenic E. coli strains in the drinking water intakes highlights the risk to human health associated with direct and indirect consumption of untreated surface water. F ecal contamination is the primary contributor of pathogens to surface water and constitutes a significant public health risk. In Canada, the estimated rate of acute gastrointestinal illness attributable to water is 0.11 cases per person per year, which poses a substantial economic burden (1). Traditionally, the safety of water used for drinking and recreation as required by regulation is assessed in terms of the concentration of Escherichia coli, a dominant intestinal inhabitant of humans and warm-blooded animals, whose presence in water indicates fecal contamination and the potential presence of enteric pathogens (2). The sources of E. coli in the aquatic environment can originate from runoff from land applied with animal wastes or animal feeding operations, urban runoff, inadequate or failing septic or sewer systems, and wildlife (3). Identification of the source of fecal pollution is a high priority in order to protect and manage source water quality and also to assess the potential public health risk associated with fecal contamination from a given host source (4). To this end, many advances have been made in recent years to develop various ...