Graduate Thesis Or Dissertation
 

Optimization of Wastewater Sampling and Concentration Methods to Accurately Detect and Quantify SARS-CoV-2 in Varying Scales of Catchment

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/xs55mm00j

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  • At the beginning of the COVID-19 pandemic, the world was unprepared for such a massive challenge to global health. Individual testing kits and personal protective equipment were scarce. There were minimal ways to track the rapid spread of the disease, until researchers discovered that SARS-CoV-2, the virus that causes COVID-19, is shed in the human stool. Wastewater-based epidemiology (WBE) was implemented across the world to track community health through a single pooled sample. Due to the rapid implementation of WBE programs, many different protocols for sampling and analysis were developed with limited knowledge on their effect on SARS-CoV-2 detection and quantification. To better understand how these methodologies affect the recovery of the SARS-CoV-2 signal, a variety of studies were conducted to evaluate different sampling and viral concentration methods. Two of the most common sampling types utilized for WBE, grab and composite, were evaluated by comparing the SARS-CoV-2 concentrations in 24 hourly grab samples with their associated 24-h composite. Additionally, the SARS-CoV-2 concentrations at a low-flow building site with high frequency (5-min and 15-min) sampling were evaluated to quantify the effects of sampling frequency on composite values. The results indicated that composite sampling should be utilized in almost every scale of catchment except for a high-flow wastewater treatment plant influent, where it may be acceptable to use grab samples if only attempting to determine the presence or absence of SARS-CoV-2. At the scale of micro-sewersheds or building effluents, high-frequency composite sampling is required to capture the temporal variation in viral signal. Another critical step to optimize is the concentration of SARS-CoV-2 from wastewater. Six concentration methods commonly used in WBE were compared using a wastewater treatment plant influent sample with a high SARS-CoV-2 concentration. The best relative recovery was found in the methods that utilized electronegative membrane filtration. To supplement this knowledge, the different solid and liquid fractions of an influent were separated into the individual components (i.e., liquid, settleable solids and non-settleable solids) to evaluate where SARS-CoV-2 RNA was partitioned in the wastewater. About 90% of total SARS-CoV-2 signal obtained in electronegative membrane filtration was found in the non-settleable solid fraction, indicating that future concentration method development should focus on retaining the SARS-CoV-2 RNA from the non-settleable solids.
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  • This work was supported by the National Science Foundation RAPID #2027679. Clean Water Services participated in this research with partial funding from the CARES Act through the Washington County Cities and Special District Assistance program.
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  • Pending Publication
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  • 2021-06-12 to 2022-07-12

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