Grantee Research Project Results
2023 Progress Report: Human Enteric Viruses and Viral Surrogates as Measures of Water Reuse Potential from Centralized and Decentralized Wastewater Treatment
EPA Grant Number: R840259Title: Human Enteric Viruses and Viral Surrogates as Measures of Water Reuse Potential from Centralized and Decentralized Wastewater Treatment
Investigators: Heufelder, George , Baumgaertel, Brian , Olmsted, Emily Michele , Pancorbo, Oscar C , Wigginton, Sarah
Institution: Barnstable County Department of Health and Environment and Massachusetts Department of Environmental Protection
EPA Project Officer: Ludwig-Monty, Sarah
Project Period: August 1, 2021 through July 31, 2024
Project Period Covered by this Report: August 1, 2022 through July 31,2023
Project Amount: $1,239,655
RFA: Viral Pathogen and Surrogate Approaches for Assessing Treatment Performance in Water Reuse (2021) RFA Text | Recipients Lists
Research Category: Water Treatment
Objective:
The aim of the present study is to compare the occurrence and log reduction values (LRV) of enteric viruses and viral surrogates in treated wastewater. Centralized and decentralized treatment modalities are evaluated for the potential for wastewater reuse following the removal of contaminants, as well as the viability of five viral surrogates as possible indicators for the presence of human enteric viruses. Testing is conducted via molecular and cultured analytical methods.
Progress Summary:
This project is successfully leveraging the assets and talents of two regional agencies: Massachusetts Alternative Septic System Test Center (MASSTC) for the full-scale test locations of candidate technologies, collection of samples and processing of culturable bacteriophage indicators, and Wall Experiment Station of Massachusetts Department of Environmental Protection (MassDEP-WES) for development of state-of-the-art assay techniques for human enteric virus, advanced instrumentation, and laboratory expertise to assay samples for human enteric virus genomic material.
• To date, over 130 sampling events have been conducted. Over 1000 assays for male-specific and somatic coliphage, and indicator bacteria have been completed with associated field measurements and chemistry measurements at MASSTC. We continue to supplement incoming wastewater with sodium bicarbonate and monitor alkalinity levels due to results indicating a viral inhibiting matrix effect caused by depleted alkalinity.
• MASSTC has added two additional technologies to be sampled. The first addition was the “toilet of the future” installed at Cape Cod Community College in Barnstable Massachusetts. This is a closed-loop toilet system that employs electrodes to self-chlorinate and recycle flush water. A HomeBiogas system, designed to digest organic waste into usable methane gas, was installed at MASSTC in August 2022 and added to this project in October 2022.
• Over 1000 samples for PCR analyses have been successfully sampled, concentrated, kept at -80oC and transported to MassDEP-WES.
• The MassDEP-WES ddPCR methods employ two analytical surrogates, bovine herpesvirus (BoHV, the DNA surrogate) and bovine coronavirus (BCoV, the RNA surrogate), spiked into all test samples, including QC samples, prior to nucleic acid extraction and carried through ddPCR analysis. These surrogates are also being spiked into a method blank and test sample (matrix spike) with each batch of test samples concentrated by ultrafiltration or centrifugal filtration in the field.
• MassDEP-WES staff has been working on the development and validation of ddPCR analytical methods for this project and associated SOPs for nucleic acid extraction and ddPCR analysis using the Bio-Rad platform. The final methods include extensive quality control elements at each of the preparation and analytical steps. All ddPCR method validation work will be completed by the end of October 2023 and analysis of project test samples will begin in November 2023.
• MassDEP-WES staff, using environmental samples, has compared the analytical performance of Bio-Rad’s ddPCR and Qiagen’s digital PCR (dPCR) platforms and found that both methods provide similar detection limits but ddPCR provides higher analytical resolution. Therefore, BioRad’s ddPCR methods will be used to quantitate all viral and viral indicator targets for this project, including indigenous human adenoviruses, human noroviruses GI and GII, and the viral indicators, crAssphage, pepper mild mottle virus (PMMoV), and the human-associated fecal marker Bacteroides HF183.
Future Activities:
In the subsequent reporting period, MASSTC will complete all sampling events and associated field measurements for all on-site and centralized wastewater treatment technologies and transport them all to WES for molecular analysis.
MassDEP-WES will complete all ddPCR and RT-ddPCR assay validations for each of the six target microbes and the two viral vaccine (RNA and DNA) positive controls. MassDEP-WES staff will complete all method development, optimization, automation, and validation, and quality assurance review and approval of all associated standard operating procedures (SOPs) by the end of October 2023. These will include an automated as well as a manual nucleic acid extraction, and ddPCR method. Starting in November 2023, MassDEP-WES staff will begin analyzing project test samples that were or will be submitted by MASSTC, processed, and frozen at - 80°C, using the optimized validated methods.
Journal Articles on this Report : 1 Displayed | Download in RIS Format
Other project views: | All 1 publications | 1 publications in selected types | All 1 journal articles |
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Type | Citation | ||
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Pfannerstill E, Arata C, Zhu Q, Schulze B, Woods R, Harkins C, Schwantes R, Mcdonald B, Goldstein A. Comparison between Spatially Resolved Airborne Flux Measurements and Emission Inventories of Volatile Organic Compounds in Los Angeles. ENVIRONMENTAL SCIENCE AND TECHNOLOGY 2023;57(41):15533-15545. |
R840259 (2023) R840010 (2022) |
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Supplemental Keywords:
Innovative/Alternative Technologies, onsite septic systems, bioreactors, EPA Region 1, soil treatment, media filters, human enteric viral pathogens, viral surrogatesProgress 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.