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Use of Plaque Assay to Detect Enteric Viruses in a Rural Watershed

Dep. of Biology, Grove City College, 100 Campus Drive, Grove City, PA 16127;
Dep. Chemistry, Chemical Engineering and Environmental Science, New Jersey Inst. of Technology, Newark, NJ.

^* Corresponding author (fjbrenner{at}gcc.edu).

ABSTRACT

Water samples were collected from four locations within the Munnell Run Watershed in Mercer County, Pennsylvania, and analyzed for fecal coliforms by MPN and enteric phages by plaque assay using Salmonella typhimurium WG 49 and Bacteroides fragilis HSP 40 as hosts. Fecal coliform concentrations and the number of phages varied seasonally (P < 0.001), as well as among the different sampling stations (P < 0.001). At all sampling stations positive for phages, S. typhimurium WG 49 PFUs outnumbered B. fragilis HSP 40 PFUs. Phages also were isolated from a septic discharge pipe and a wetland receiving septic drainage, but not from three avian species, 12 mammalian species, two streams without human wastes or from natural wetland, indicating that these viruses were of human origin. MPNs of fecal coliforms and S. typhimurium and B. fragilis PFUs were correlated with stream temperature (P < 0.001) and rainfall (P < 0.01). Only fecal coliforms and S. typhimurium WG 49 phages were correlated with suspended solids concentrations (P < 0.01). Likewise, there was a significant interaction among these parameters and MPNs of fecal coliform and HSP 40 and WG 49 PFUs (P < 0.001). The presence of host specific phages indicate the existence of septic discharges in the watershed, but both fecal coliforms and enteric viruses persist in stream systems, especially during the summer months.

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