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Multi-laboratory evaluation of methods to detect coliphages in large volumes of ground water



Multi-laboratory evaluation of methods to detect coliphages in large volumes of ground water



Abstracts of the General Meeting of the American Society for Microbiology 102: 437



Two new methods (1601 and 1602), to detect coliphages as virus indicators of fecal contamination in large (0.1-1 liter) volumes of groundwater were evaluated in samples seeded with mixed populations of coliphages from sewage. Three strains or E. coli bacterial hosts were used: CN13 for somatic coliphages, Famp for F+ coliphages and C3000 for both somatic and F+ coliphages. A single agar layer plaque assay method, EPA Method 1602, resulted in moderate (apprx35%) to excellent (apprx85%) detection of apprx100 coliphages in 0.1-liter groundwater samples, regardless of E. coli host. Detected plaques were confirmed by picking and re-spotting onto host bacteria lawns with success rates of 57 to 99%. When 1-L volumes of ground water seeded with 1-2 PFU of coliphages were analyzed by two-step enrichment method (EPA Method 1601), there was a very high likelihood of coliphage detection, with average recoveries of 30 to >100%. Variability in coliphage recoveries, based on calculated 95% confidence intervals, were within the expected range for an MPN assay. Of 82 total trials in which 10 1-L bottles of ground water were seeded with 1-2 coliphages/L, detection was achieved in 79 trials (96%). Hence, there is a very high probability of detecting low levels of coliphages in 1-liter volumes of groundwater using this method. In comparing the two methods, participating labs favored the enrichment over the single agar layer method, the former being easier to perform, sensitive in detecting low numbers of coliphages and more consistent in its results. By the enrichment method, both somatic and male-specific coliphages were detected simultaneously on a single host, E. coli C3000, at sensitivities comparable to detecting either somatic or male-specific coliphages individually. The simultaneous detection of both somatic and male-specific coliphages simplifies the method as well as lowers costs. Enrichment Method 1601 appears to be efficient and sensitive for detecting low levels of coliphages in ground water.

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Accession: 035349271

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