Grantee Research Project Results

2024 Progress Report: A cross-regional, cross-laboratory investigation of viral pathogens and surrogates in wastewater for non-potable reuse

EPA Grant Number: R840257
Title: A cross-regional, cross-laboratory investigation of viral pathogens and surrogates in wastewater for non-potable reuse
Investigators: Jiang, Sunny C. , Goel, Ramesh , Yan, Tao , Solo-Gabriele, Helena , Sherchan, Samendra , Whiteson, Katrine , Rosso, Diego , Bischel, Heather N
Institution: University of California - Irvine , University of California Davis , University of Miami , Tulane University , University of Hawaii, Manoa , University of Utah
EPA Project Officer: Ludwig-Monty, Sarah
Project Period: August 1, 2021 through July 31, 2024 (Extended to July 31, 2025)
Project Period Covered by this Report: August 1, 2023 through July 31,2024
Project Amount: $1,240,000
RFA: Viral Pathogen and Surrogate Approaches for Assessing Treatment Performance in Water Reuse (2021) RFA Text | Recipients Lists
Research Category: Water Treatment

Objective:

1) identify a suite of viral pathogens and surrogates, and their detection methods that may best serve as the indication of viral risk in water reuse; 2) compare viral pathogen and surrogate detection methods in wastewater by different labs in order to develop a set of Standard Operational Protocols (SOP) that can be adopted as standard methods by water utilities; 3) apply the SOPs to wastewater samples collected from wastewater reclamation plants across the U.S., from Miami to Hawaii, at different seasons and treatment processes to identify the best markers for viral risk in water reuse; 4) develop Artificial Neural Network (ANN) models to predict infectious viruses in the effluent using pathogen/surrogate and treatment conditions; and 5) integrate the outcomes from the cross-regional and cross-laboratory study into quantitative microbial risk assessment (QMRA) models to estimate health risk from exposure to the water for non-potable reuse.

Progress Summary:

Samples were collected and analyzed for coliphage (somatic and F-specific) infectivity for 1.5 year from 6 wastewater utilities quarterly.
Viruses in samples were concentrated using two standardized methods (PEG precipitation & e-negative filtration).
Water quality and physical/chemical parameters were collected for all sample collection dates.
Drafted manuscript on phage removal and cross regional comparison of between water quality and virus log removals.
Extracted nucleic acids from all virus concentrates from all samples.
Developed standardized protocols for target viral assays by qPCR and ddPCR and completed 80% of the sample analysis
Developed and optimized flow cytometry assay methods for quantification of virus-like particles in wastewater by UC Davis team.
Carried out preliminary Machine Learning Analysis to predict the virus log removal

Future Activities:

Complete the manuscript for coliphage cross regional comparison for submission
Perform statistical analysis to identify the relationships between viral surrogates and seasonal, geographical locations as well as WWTPs characteristics.
Analysis data from field data collection for viral removal efficiency by direct counting of virus-like particles using flow cytometry as a rapid measurement of viral removal in two wastewater treatment plants by UC Davis and UC Irvine.
Complete all qPCR, ddPCR and metagenomic analysis of viral concentrates from all seasonal and plant collections across the U.S.
Complete of Artificial Neural Network and perform the analysis between wastewater characteristics and viral surrogates.
Carried out Quantitative Microbial Rish Analysis
Compete the manuscript for ddPCR analysis of pathogenic viruses

Journal Articles:

No journal articles submitted with this report: View all 6 publications for this project

Supplemental Keywords:

coliphage, infectivity, viral concentration, genetic analysis, Flow Cytometry, artificial neural network, quantitative microbial risk assessment

Progress and Final Reports:

Original Abstract

The 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.