Grantee Research Project Results
2024 Progress Report: Novel Quantitative Methods for Indigenous Viruses in Wastewater, Improving the Assessment of Water Reuse Treatment Performance
EPA Grant Number: R840258Title: Novel Quantitative Methods for Indigenous Viruses in Wastewater, Improving the Assessment of Water Reuse Treatment Performance
Investigators: Gim Aw, Tiong , Rose, Joan B. , Shen, Yun
Institution: Tulane University , University of California Riverside , Michigan State University
EPA Project Officer: Ludwig-Monty, Sarah
Project Period: August 1, 2021 through July 31, 2024 (Extended to July 31, 2026)
Project Period Covered by this Report: August 1, 2023 through July 31,2024
Project Amount: $1,239,241
RFA: Viral Pathogen and Surrogate Approaches for Assessing Treatment Performance in Water Reuse (2021) RFA Text | Recipients Lists
Research Category: Water Treatment
Objective:
Project objectives are to: (1) characterize the composition of indigenous viruses in wastewater (including viruses with three different forms: free, particle-associated, and vesicle-cloaked viruses) using a metagenomic approach, with the purpose of identification of viral surrogates for assessing treatment performance; (2) design and develop individual based quantitative assays for new viral surrogates using new instrumentation and molecular viability methods; and (3) validate new surrogates and new quantitative detection assays (from Objective 1 & 2) for estimating reduction of viruses through full-scale advanced water treatment processes (with some bench-scale work).
Progress Summary:
We successfully collected wastewater samples from the Upper Occoquan Service Authority (UOSA) wastewater treatment plant and completed monitoring of both human noroviruses (HuNoV) and HuNoV vesicles at each stage of the treatment process. Our results showed as the treatment process progresses, the overall amount of HuNoV decreases. However, the proportion of viral vesicles relative to the total number of viruses increases. This suggests that viral vesicles have a higher survival rate compared to free viruses during the wastewater treatment process. The vesicles appear to protect the enclosed viruses from certain disinfection and treatment methods, which may have important implications for public health and wastewater treatment practices.
We also completed metagenomic sequencing for particle-associated viruses in secondary treated wastewater. We collected secondary treated wastewater samples (n = 30) from five full-scale water reuse facilities in California, Florida and Ohio. Nearly complete genomes were assembled from wastewater samples including a novel water quality indicator, tomato brown rugose fruit virus. This study provides insights on particles-associated viruses in secondary treated wastewater which is the source water for reclamation. A robust metagenomic analysis can guide the development of novel viral indicators and surrogates for assessing the performance of wastewater treatment and water reuse processes.
We evaluated the efficiency of intercalating azo dyes propidium monoazide (PMA) for the quantification of viable viruses in wastewater and water samples using digital PCR. The results suggested the potential of a combined viability assay and digital PCR approach for directly quantifying virus removal and inactivation in full-scale wastewater and water treatment systems.
Future Activities:
We will continue wastewater sampling from full-scale water reuse systems to isolate free, particle-associated, and vesicle-cloaked viruses for metagenomic sequencing to determine changes in virus diversity after each treatment step. We will apply viability digital PCR assays for quantifying virus removal during full-scale advanced water treatment processes. We will conduct bench-top experiments to study the mechanisms and removal efficiencies of particle-associated viruses and viral vesicles through disinfection.
Journal Articles:
No journal articles submitted with this report: View all 8 publications for this projectSupplemental Keywords:
Pathogens, effluent, discharge, innovative technology, analytical, measurement methods, monitoring, geneticsProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.