Abstract
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 environment. Reported here are novel and effective laboratory protocols for viral concentration and highly sensitive and optimized RT-PCR for the efficient detection of enteroviruses, an important enteric virus subset, in Hawaiian environmental waters. Eighteen published enterovirus primer pairs were comparatively evaluated for detection sensitivity. The primer set exhibiting the lowest detection limit under optimized conditions, EQ-1/EQ-2, was validated 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 at these locations. As an additional means of viral concentration, shellfish were collected from 9 sample sites and subjected to dissection, RNA extraction, and subsequent RT-PCR. Shellfish tissue from 6 of 9 sites tested positive for enterovirus. The techniques implemented here are valuable resources to aid accurate reflection of microbial contamination in Hawaii’s environmental waters.
Highlights
Sewage-contaminated recreational water can pose numerous health risks to the public; effective water quality monitoring is absolutely essential [1]
These indicators often fail to reflect the presence of important hazardous viruses [2]. This is of important concern, as viral pathogens shed in human feces may compromise public safety by polluting recreational waters that meet bacterial indicator standards
Because reliance on bacterial indicators alone for water quality surveillance fails to reflect the presence of potentially problematic viral pathogens, a need for alternative monitoring parameters exists [2,3,4]
Summary
Sewage-contaminated recreational water can pose numerous health risks to the public; effective water quality monitoring is absolutely essential [1]. Microbiological water quality is primarily assessed via bacterial indicators such as enterococci, fecal coliform, and total coliform bacteria These indicators often fail to reflect the presence of important hazardous viruses [2]. This is of important concern, as viral pathogens shed in human feces may compromise public safety by polluting recreational waters that meet bacterial indicator standards. These bacterial indicators may grow naturally in tropical environments, resulting in inaccurate assessment of water pollution levels [3]. Alternative monitoring systems are needed to improve the surveillance of recreational waters and secure public protection from waterborne disease [4]
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