Comparison of methods for the detection of coliphages in recreational water at two California, United States beaches

Methods for detection of two fecal indicator viruses, F+ and somatic coliphages, were evaluated for application to recreational marine water. Marine water samples were collected during the summer of 2007 in Southern California, United States from transects along Avalon Beach (n = 186 samples) and Doheny Beach (n = 101 samples). Coliphage detection methods included EPA method 1601 – two-step enrichment (ENR), EPA method 1602 – single agar layer (SAL), and variations of ENR. Variations included comparison of two incubation times (overnight and 5-h incubation) and two final detection steps (lysis zone assay and a rapid latex agglutination assay). A greater number of samples were positive for somatic and F+ coliphages by ENR than by SAL (p < 0.01). The standard ENR with overnight incubation and detection by lysis zone assay was the most sensitive method for the detection of F+ and somatic coliphages from marine water, although the method takes up to three days to obtain results. A rapid 5-h enrichment version of ENR also performed well, with more positive samples than SAL, and could be performed in roughly 24 h. Latex agglutination-based detection methods require the least amount of time to perform, although the sensitivity was less than lysis zone-based detection methods. Rapid culture-based enrichment of coliphages in marine water may be possible by further optimizing culture-based methods for saline water conditions to generate higher viral titers than currently available, as well as increasing the sensitivity of latex agglutination detection methods.

► Methods for detection of coliphages in recreational marine waters were evaluated.
► The EPA1601 was best and a short version of EPA1601 was the second best at detecting coliphages.
► The use of CLAT showed lower sensitivity when compared to spot-plating at detecting coliphages.