Candidate biomarkers of antibiotic resistance for the monitoring of wastewater and the downstream environment

A Margarida Teixeira; Ivone Vaz-Moreira; David Calderón-Franco; David Weissbrodt; Sabina Purkrtova; Stanislav Gajdos; Giulia Dottorini; Per Halkjær Nielsen; Leron Khalifa; Eddie Cytryn; Jan Bartacek; Célia M Manaia

doi:10.1016/j.watres.2023.120761

Candidate biomarkers of antibiotic resistance for the monitoring of wastewater and the downstream environment

Water Res. 2023 Dec 1:247:120761. doi: 10.1016/j.watres.2023.120761. Epub 2023 Oct 19.

Affiliations

¹ CBQF - Centro de Biotecnologia e Química Fina - Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua de Diogo Botelho 1327, Porto 4169-005, Portugal.
² Department of Biotechnology, Environmental Biotechnology Section, Delft University of Technology, van der Maasweg 9, Delft, HZ 2629, the Netherlands.
³ Department of Biotechnology, Environmental Biotechnology Section, Delft University of Technology, van der Maasweg 9, Delft, HZ 2629, the Netherlands; Department of Biotechnology and Food Science, Norwegian University of Science and Technology, Trondheim 7034, Norway.
⁴ Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, 5 Technická, Prague 166 28, Czech Republic.
⁵ Department of Water Technology and Environmental Engineering, Faculty of Environmental Technology, University of Chemistry and Technology Prague, 5 Technická, Prague 166 28, Czech Republic.
⁶ Department of Chemistry and Bioscience, Center for Microbial Communities, Aalborg University, Aalborg 9220, Denmark.
⁷ Institute of Soil, Water and Environmental Sciences, The Volcani Institute, Agricultural Research Organization, P.O Box 15159, Rishon Lezion 7528809, Israel.
⁸ CBQF - Centro de Biotecnologia e Química Fina - Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua de Diogo Botelho 1327, Porto 4169-005, Portugal. Electronic address: cmanaia@ucp.pt.

PMID: 37918195
DOI: 10.1016/j.watres.2023.120761

Abstract

Urban wastewater treatment plants (UWTPs) are essential for reducing the pollutants load and protecting water bodies. However, wastewater catchment areas and UWTPs emit continuously antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), with recognized impacts on the downstream environments. Recently, the European Commission recommended to monitor antibiotic resistance in UWTPs serving more than 100 000 population equivalents. Antibiotic resistance monitoring in environmental samples can be challenging. The expected complexity of these systems can jeopardize the interpretation capacity regarding, for instance, wastewater treatment efficiency, impacts of environmental contamination, or risks due to human exposure. Simplified monitoring frameworks will be essential for the successful implementation of analytical procedures, data analysis, and data sharing. This study aimed to test a set of biomarkers representative of ARG contamination, selected based on their frequent human association and, simultaneously, rare presence in pristine environments. In addition to the 16S rRNA gene, ten potential biomarkers (intI1, sul1, ermB, ermF, aph(3'')-Ib, qacEΔ1, uidA, mefC, tetX, and crAssphage) were monitored in DNA extracts (n = 116) from raw wastewater, activated sludge, treated wastewater, and surface water (upstream and downstream of UWTPs) samples collected in the Czech Republic, Denmark, Israel, the Netherlands, and Portugal. Each biomarker was sensitive enough to measure decreases (on average by up to 2.5 log-units gene copy/mL) from raw wastewater to surface water, with variations in the same order of magnitude as for the 16S rRNA gene. The use of the 10 biomarkers allowed the typing of water samples whose origin or quality could be predicted in a blind test. The results show that, based on appropriate biomarkers, qPCR can be used for a cost-effective and technically accessible approach to monitoring wastewater and the downstream environment.

Keywords: Anthropogenic pollution; Antibiotic resistance; Gene monitoring; Quantitative PCR; Wastewater.

MeSH terms

Angiotensin Receptor Antagonists / analysis
Angiotensin-Converting Enzyme Inhibitors / analysis
Anti-Bacterial Agents / analysis
Anti-Bacterial Agents / pharmacology
Drug Resistance, Microbial / genetics
Genes, Bacterial*
Humans
RNA, Ribosomal, 16S / genetics
Wastewater*
Water / analysis

Substances

Wastewater
RNA, Ribosomal, 16S
Angiotensin Receptor Antagonists
Angiotensin-Converting Enzyme Inhibitors
Anti-Bacterial Agents
Water