<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.
(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>