Impact of enumeration method on diversity of Escherichiacoli genotypes isolated from surface water.
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UNLABELLED: There are numerous regulatory-approved Escherichiacoli enumeration methods, but it is not known whether differences in media composition and incubation conditions impact the diversity of E.coli populations detected by these methods. A study was conducted to determine if three standard water quality assessments, Colilert , USEPA Method 1603, (modified mTEC) and USEPA Method 1604 (MI), detect different populations of E.coli. Samples were collected from six watersheds and analysed using the three enumeration approaches followed by E.coli isolation and genotyping. Results indicated that the three methods generally produced similar enumeration data across the sites, although there were some differences on a site-by-site basis. The Colilert method consistently generated the least diverse collection of E.coli genotypes as compared to modified mTEC and MI, with those two methods being roughly equal to each other. Although the three media assessed in this study were designed to enumerate E.coli, the differences in the media composition, incubation temperature, and growth platform appear to have a strong selective influence on the populations of E.coli isolated. This study suggests that standardized methods of enumeration and isolation may be warranted if researchers intend to obtain individual E.coli isolates for further characterization. SIGNIFICANCE AND IMPACT OF THE STUDY: This study characterized the impact of three USEPA-approved Escherichiacoli enumeration methods on observed E.coli population diversity in surface water samples. Results indicated that these methods produced similar E.coli enumeration data but were more variable in the diversity of E.coli genotypes observed. Although the three methods enumerate the same species, differences in media composition, growth platform, and incubation temperature likely contribute to the selection of different cultivable populations of E.coli, and thus caution should be used when implementing these methods interchangeably for downstream applications which require cultivated isolates.
