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Grantee Research Project Results

Final Report: Prevalence, abundance, and fate of antibiotics, antibiotic-resistant bacteria, and their determinant genes in U.S. wastewater systems

EPA Grant Number: R840826
Title: Prevalence, abundance, and fate of antibiotics, antibiotic-resistant bacteria, and their determinant genes in U.S. wastewater systems
Investigators:
Institution:
EPA Project Officer:
Project Period: September 1, 2024 through May 7, 2025
Project Amount: $2,350,211
RFA: National Priorities: Evaluation of Antimicrobial Resistance in Wastewater and Sewage Sludge Treatment and Its Impact on the Environment Request for Applications (RFA) (2023) RFA Text |  Recipients Lists
Research Category: Human Health , Water , Water Treatment

Objective:

To quantify the fate of antibiotic-resistant bacteria, antibiotic resistance genes, and antibiotics in wastewater treatment utilities across the U.S.

Summary/Accomplishments (Outputs/Outcomes):

This project advanced the development of a national wastewater surveillance framework for antimicrobial resistance (AMR) markers, generating foundational data and infrastructure for future public health monitoring. Key outputs and outcomes include:

  • Wastewater Surveillance Infrastructure & Sampling
    • Successful enrollment of 40 wastewater utilities across five geographic regions, representing diverse plant sizes and treatment configurations.
    • Completion of winter 2025 sampling campaign across all sites, with samples collected from influent, effluent stages, raw sludge, and biosolids.
    • Standardized protocols for microbial, molecular, and chemical analyses were developed and implemented under an EPA-approved QAPP.
  • Laboratory & Analytical Outputs
    • Culture-based data generated for four microbial targets: Escherichia coli, extended-spectrum beta-lactamases (ESBL) E. coli, enterococci, and vancomycin-resistant enterococci (VRE).
    • Physical-chemical parameters analyzed with QA/QC verification.
    • Molecular and chemical samples processed and archived for future analysis (e.g., DNA extraction, dPCR, mass spectrometry), though further processing was halted due to funding conclusion.
  • Capacity Building & Training
    • Three PhD students onboarded and trained in environmental microbiology, analytical chemistry, and stakeholder engagement.
    • Interdisciplinary skill development supported future leadership in wastewater surveillance and public health research.
    • Postdoctoral researcher recruited to lead molecular resistome analysis, though work was not initiated due to early funding conclusion.
  • Stakeholder Engagement & Dissemination
    • Collaborative relationships established with utility partners, many expressing interest in continued engagement.
    • Preliminary findings presented at the 2025 Association of Environmental Engineering and Science Professors Conference, highlighting regional trends and utility-level variability.
    • Anonymized data summaries shared with utility collaborators to support local decision-making.
    • Planning initiated for peer-reviewed publications and broader public outreach, contingent on future support.

Supplemental Keywords:

Wastewater epidemiology, AMR surveillance, utility partnerships, microbial indicators, environmental microbiology, environmental health research

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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.

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Last updated April 28, 2023
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