Grantee Research Project Results

2023 Progress Report: Advancing Safety and Reliability to Protect Public Health, Identifying Quantitative Reductions of Viral Pathogens and Surrogates for Water Reuse Applications

EPA Grant Number: R840260
Title: Advancing Safety and Reliability to Protect Public Health, Identifying Quantitative Reductions of Viral Pathogens and Surrogates for Water Reuse Applications
Investigators: Jang, Grace , Pepper, Ian L. , Reynolds, Kelly A. , Gerba, Charles P. , Achilli, Andrea , Betancourt, Walter
Institution: Water Research Foundation , University of Arizona
EPA Project Officer: Ludwig-Monty, Sarah
Project Period: August 1, 2021 through April 25, 2025
Project Period Covered by this Report: August 1, 2022 through July 31,2023
Project Amount: $1,239,813
RFA: Viral Pathogen and Surrogate Approaches for Assessing Treatment Performance in Water Reuse (2021) RFA Text | Recipients Lists
Research Category: Water Treatment

Objective:

The overall goal of this project is to improve public health protection through the development of quantitative data on the removal of human enteric viruses by treatment processes where wastewater is the source water. This data can be used by the water sector, communities, and regulators to provide log reduction values (LRV) to meet desired risk goals.

Progress Summary:

During this reporting period, the team conducted sampling at the selected water reuse facilities. Samples were collected from various locations at the participating facility and analyzed using digital PCR (dPCR) and cell culture to detect virus genomes and culturable viruses, respectively.

Our initial findings indicate that infectious coliphages were detected throughout the entire treatment process in the sampled facilities to date. Concentrated samples (equivalent to 100 to 200 liters) were assayed for infectious coliphages, which, to the best of our knowledge, is a larger sample volume than previously assayed in any study. This allows us to assess as much as a 10-log reduction of infectious virus.

Adenovirus appears to be more resistant to treatment processes when using the A549 cell line compared to the BGM cell line. We are currently in the process of completing cell culture assays, which should provide valuable insights into the potential coliphages and adenoviruses for measurements of LRVs of viruses.

Initial data indicates infectious coliphage (Somatic Coliphage and F+ Coliphage) may be a potentially useful indicator for both assessment of Log Reduction Values (LRVs) and in monitoring treatment processes. Four significant findings to date are the following:

Infectious adenovirus and indigenous coliphages are detected after a series of treatment and disinfection processes. The degree of removal is less after the initial treatment step.
Infectious adenovirus and stomatic and F+ coliphages removal by the different treatment processes is similar.
Indigenous coliphages could potentially be used as indicators of virus removal performance by advanced treatment processes.
Increasing the number of cell culture passages from two to three results in greater isolation of infectious adenoviruses. This indicates that adenovirus removal is underestimated if only two cell passages are performed.

Future Activities:

Continue collecting sampling at participating utilities and conducting cell culture assay for human pathogenic viruses. Note that cell culture assay and confirmation of infectious virus in cell culture typically require 3-4 months to complete.
Identify viruses isolated in cell culture.
Continue conducting engineering scale tests to validate potential LRV under controlled conditions.

Journal Articles:

No journal articles submitted with this report: View all 1 publications for this project

Supplemental Keywords:

Water Reuse, Risk Assessment, Viruses, Surrogate, Treatment

Relevant Websites:

WRF 4955 - Indicator Viruses for Advanced Physical Treatment Process Performance Confirmation Exit

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.