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Genome-resolved metagenomics uncovers antimicrobial resistance gene carriers in hospital and municipal wastewater environments

Silvester, Reshma, Webster, Gordon ORCID: https://orcid.org/0000-0002-9530-7835, Perry, William B., Farkas, Kata, Rushton, Laura, Craine, Noel, Cross, Gareth, Kille, Peter ORCID: https://orcid.org/0000-0001-6023-5221, Weightman, Andrew J. ORCID: https://orcid.org/0000-0002-6671-2209 and Jones, Davey L. 2025. Genome-resolved metagenomics uncovers antimicrobial resistance gene carriers in hospital and municipal wastewater environments. Science of the Total Environment 1002 , 180607. 10.1016/j.scitotenv.2025.180607

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Abstract

Wastewater-based epidemiology (WBE) is a powerful approach to study antimicrobial resistance (AMR) dynamics at the population level. Using genome-resolved metagenomics, we recovered 3978 metagenome-assembled genomes (MAGs) from archived metagenome sequences generated under the national wastewater surveillance programme across Wales, UK. Taxonomic profiling of MAGs revealed a diverse bacterial community, with significant compositional shifts observed across seasons and sample sources. Approximately 13.6 % of the MAGs carried one or more antimicrobial resistance genes (ARGs), with genes conferring resistance to tetracycline and oxacillin being the most prevalent within the wastewater microbiomes. We also recovered yet-uncultivated microbial genomes- often referred to as “microbial dark matter” harbouring clinically relevant ARGs, offering insights into previously uncharacterised resistance reservoirs in wastewater. ARG-host associations shifted between untreated influent and treated effluent, with effluent profiles also varying significantly between secondary and tertiary treatments, revealing the impact of treatment type on ARG host composition. This study represents the first comprehensive genome-resolved metagenomic characterisation of ARG carriers across both hospital and municipal wastewater in Wales, effectively bridging clinical and environmental compartments. Our findings highlight the need to integrate high-resolution genome-resolved metagenomic surveillance into national AMR monitoring frameworks to track emerging threats, characterise ARG reservoirs and inform targeted public health interventions.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Schools > Biosciences
Schools > Pharmacy
Publisher: Elsevier
ISSN: 0048-9697
Date of First Compliant Deposit: 6 October 2025
Date of Acceptance: 23 September 2025
Last Modified: 06 Oct 2025 15:15
URI: https://orca.cardiff.ac.uk/id/eprint/181499

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