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<p>(1) Determine infective /total particle ratio in fresh human stool for G1. 1 and GII.4 stool samples using the PGM-MB/RT-PCR. Determine infective /total norovirus particles in wastewater treatment (WWT) and lagoon influent and effluent. Evaluate the effect of chlorination, and UV as used in WWT.
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(2) Evaluate impact of bio accumulation on infective/total particle ratio in oyster as compared to harvest water. Evaluate the effect of sunlight and other environmental conditions on viability of norovirus. (3) In conjunction with objectives 1 and 2, the effect of WWT/oyster bioaccumulation and environmental conditions will be evaluated on male-specific coliphage. Quantitative measures of the number of male-specific MS2 (phage MS2) particles by qRT-PCR and conventional culture techniques. Phage MS2 is an enteric viral surrogate for human norovirus.</p>
<p>Viability Assays. PGM-MB assay will be performed as previously described (Dancho et al 2012, IJFM 155:222.) after UV, chlorine, sunlight, or other treatments of norovirus (essentially as described in Kingsley et al 2014 IJFM 171:94.) Experiments will be performed in triplicate (N=3; n=9). Male-specific coliphage assays will be performed on WWT samples, shellfish, and stool samples to determine persistence of this sewage indicator relative to human norovirus viability.</p>