Connell, Christina Noelle2016-02-192016-02-192012-05http://hdl.handle.net/10125/101292M.S. University of Hawaii at Manoa 2012.Includes bibliographical references.Health risks associated with sewage-contaminated recreational waters are of important public health concern. Reliable water monitoring systems are therefore crucial. Current recreational water quality criteria rely predominantly on the enumeration of bacterial indicators, while potentially dangerous viral pathogens often remain undetected. Human enteric viruses have been proposed as alternative indicators; however, their detection is often hindered by low viral concentrations present in the aquatic environment. Reported here are novel and effective laboratory protocols for enhanced enteric virus detection in Hawaiian environmental waters. First, a fine-tuned, highly optimized assay for the detection of enterovirus, an important enteric virus subset, was developed by comparatively evaluating eighteen published enterovirus primer pairs for detection sensitivity. The primer set exhibiting the lowest detection limit under optimized conditions, EQ-1/EQ-2, was validated through testing urban wastewater, and then utilized in a field survey of 22 recreational bodies of water located around the island of Oahu, Hawaii. Eleven sites tested positive for enterovirus, indicating fecal contamination in a significant portion of Hawaiian waters. Additionally, the filter-feeding phenomenon of indigenous bivalve mollusks was explored as a natural bioconcentration technique to infer microbial quality of the surrounding waters. Shellfish were collected from 12 coastal locations and dissected for subsequent nucleic acid extraction from internal tissues. Optimized RT-PCR/PCR protocols were then applied to test for the presence of various enteric viruses, including enterovirus, adenovirus, norovirus genogroups I and II, and F-specific RNA coliphage. Shellfish collected from around the island tested positive for several enteric virus types, indicating that these animals are indeed natural and competent bioindicators of water quality. The extremely sensitive and innovative techniques implemented here are valuable resources to aid accurate reflection of microbial contamination in Hawaii's environmental waters.engmicrobiologyhuman enteric virusesHawaiiThesis