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Advancements in Virus Detection and Water Quality Monitoring: Insights from Freshwater and Wastewater Studies

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Gouthro, Madison

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Abstract

Human viruses are present in both wastewater and freshwater. In wastewater, they are difficult to effectively treat and therefore can end up in our freshwater systems. These viruses pose a significant health risk in freshwater environments however, current monitoring methods are inadequate for efficiently detecting their presence. Therefore, robust viral surveillance methods are essential to detect the presence of viruses in these waters, ensuring these waters remain safe for human and ecological health. To address the challenges associated with conventional sampling techniques, passive sampling has emerged as a promising alternative for viral concentration. In wastewater, wastewater-based epidemiology is commonly used to provide insights into the health of served communities where in freshwater, viral surveillance efforts have predominantly focused on the detection of enteric pathogens. This thesis addresses the pressing need for improved viral concentration and detection methodologies, with a focus on respiratory viruses in freshwater and wastewater environments. Through the assessment of various total nucleic acid extraction kits and elution buffers, this research aims to refine passive sampling protocols using granular activated carbon (GAC)-based samplers to enhance the detection and quantification of respiratory viral RNA. Field applications demonstrated the effectiveness of GAC-based passive samplers in capturing SARS-CoV-2, INFA, RSV, and MeV in freshwater, significantly enhancing the sensitivity and reliability of viral surveillance. Furthermore, this research explores the occurrence of viral genes for the same four respiratory viruses in relation to various physical water quality characteristics and extreme climate events in recreational freshwater environments. Through the deployment of GAC-based passive samplers, findings indicated high viral abundance in highly recreational lakes, with major climate events like wildfires and rainstorms further impacting viral contamination levels. Finally, this research introduces a rapid viral concentration method using powdered activated charcoal sodium alginate (PAC-NaA) hydrogel beads, optimized for capturing viruses from wastewater. The study successfully detected endogenous SARS-CoV-2 and Adenovirus using the PAC-NaA hydrogel beads. Overall, by improving these methodologies, the study aims to provide more effective and reliable tools for viral surveillance, thereby protecting public health and ensuring the safety of aquatic ecosystems.

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Water, Wastewater, RNA, Respiratory Viruses, Concentration Technique, Molecular Methods, RT-qPCR, Freshwater, Surveillance

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