Monitoring antibiotic resistance genes in wastewater environments: The challenges of filling a gap in the One-Health cycle

Miłobędzka, Aleksandra; Ferreira, Catarina; Vaz‐Moreira, Ivone; Calderón-Franco, David; Górecki, Adrian; Purkrtová, Sabina; Bartáček, Jan; Dziewit, Łukasz; Singleton, Caitlin M.; Nielsen, Per Halkjær; Weissbrodt, David G.; Manaia, Célia M.

doi:10.1016/j.jhazmat.2021.127407

Cited by 72 publications

(50 citation statements)

References 114 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rhodoferax is abundant in WWTP effluents and often identified as AMR carriers by encoding multiple efflux pumps (Jin et al, 2020; Zhou et al, 2020). ARGs have been identified from metagenome-assembled genomes (MAGs) retrieved from activated sludge from Danish WWTPs (Miłobedzka et al, 2022; Singleton et al, 2021): Rhodoferax was the most abundant population containing multiple ARGs.…”

Section: Resultsmentioning

confidence: 99%

Metagenomic profiling and transfer dynamics of antibiotic resistance determinants in a full-scale granular sludge wastewater treatment plant

Calderón-Franco

Sarelse

Christou

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

In the One Health context, wastewater treatment plants (WWTPs) are central to safeguard water resources. Nonetheless, many questions remain about their effectiveness to prevent the dissemination of antimicrobial resistance (AMR). Most surveillance studies monitor the levels and removal of selected antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in intracellular DNA (iDNA) extracted from WWTP influents and effluents. The role of extracellular free DNA (exDNA) in wastewater is mostly overlooked. In this study, we analyzed the transfer of ARGs and MGEs in a full-scale Nereda® reactor removing nutrients with aerobic granular sludge. We tracked the composition and fate of the iDNA and exDNA pools of influent, sludge, and effluent samples. Metagenomics was used to profile the microbiome, resistome, and mobilome signatures of iDNA and exDNA extracts. Selected ARGs and MGEs were analyzed by qPCR. From 2,840 ARGs identified, the genes arr-3 (2%), tetC (1.6%), sul1 (1.5%), oqxB (1.2%), and aph(3")-Ib (1.2%) were the most abundant among all sampling points and bioaggregates. Pseudomonas, Acinetobacter, Aeromonas, Acidovorax, Rhodoferax, and Streptomyces populations were the main hosts of ARGs in the sludge. In the effluent, 478 resistance determinants were detected, of which 89% from exDNA potentially released by cell lysis during aeration in the reactor. MGEs and multiple ARGs were co-localized on the same extracellular genetic contigs. These can pose a risk for AMR dissemination by transformation into microorganisms of receiving water bodies. Total intracellular ARGs decreased 3-42% as a result of wastewater treatment. However, the ermB and sul1 genes increased by 2 and 1 log gene copies mL-1, respectively, in exDNA from influent to effluent. The exDNA fractions need to be considered in AMR surveillance, risk assessment, and mitigation.

show abstract

Section: Resultsmentioning

confidence: 99%

Metagenomic profiling and transfer dynamics of antibiotic resistance determinants in a full-scale granular sludge wastewater treatment plant

Calderón-Franco

Sarelse

Christou

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…It is worth mentioning that successful PCR is not only depended on a proper primer design. It was previously described that composition of the original sample can have important implications during amplification (Miłobedzka et al, 2021). Main problem which can occur is related to difficulties during DNA purification.…”

Section: Discussionmentioning

confidence: 99%

“…Testing the occurrence of ARGs in various environments is important, but also the abundance of these genes should be analysed. For such studies the qPCR technique is used (Miłobedzka et al, 2021). Here, we developed a qPCR protocols for testing the abundance of the mcr-1 and mcr-3 genes, since they are of special clinical relevance being hosted mostly by bacterial strains isolated from hospital patients.…”

Section: Discussionmentioning

confidence: 99%

Development and validation of novel PCR primers for identification of plasmid-mediated colistin resistance (mcr) genes in various environmental settings

Górecki¹,

Musialowski²,

Wołącewicz³

et al. 2022

Journal of Hazardous Materials

Self Cite

View full text Add to dashboard Cite

Antibiotic resistance is considered one of the biggest threats to public health and has become a major concern for governments and international organizations. Combating this problem starts with improving global surveillance of antibiotic resistance genes (ARGs) and applying standardized protocols, both in a clinical and environmental context, in agreement with the One Health approach. Exceptional efforts should be directed to controlling ARGs conferring resistance to Critically Important Antimicrobials (CIA). In this study, a systematic literature review to synthesize data on the identification of mcr genes using a PCR technique was performed. Additionally, a novel set of PCR primers for mcr-1 -mcr-9 genes detection was proposed. The developed primers were in silico and experimentally validated by comparison with mcr-specific PCR primers reported in the literature. This validation, besides being a proof-of-concept for primers' usefulness, provided insight into the distribution of mcr genes in municipal wastewater, clay and river sediments, glacier moraine, manure, seagulls and auks feces and daphnids from four countries. This analysis proved that commonly used primers may deliver false results, and some mcr genes may be overlooked in tested samples. Newly-developed PCR primers turned out to be relevant for the screening of mcr genes in various environments.

show abstract

“…Hospitals are hot spots for antibiotic resistant bacteria and resistance genes. These latter, harbored either by bacterial cells, plasmids or phages, are certainly eliminated via wastewater discharge contributing to the widespread dissemination of antibiotic resistance in the environment and possibly impacting human health [ 9 ]. Monitoring the occurrence of antibiotic-resistant bacteria in hospital sewage would; therefore, appear to be of crucial importance.…”

Section: Introductionmentioning

confidence: 99%

First detection of vanA positive Enterococcus faecium clonal complex 17 in hospital wastewater in Algeria: an epidemiological report

Cherak

Bendjama

Moussi

et al. 2022

New Microbes and New Infections

View full text Add to dashboard Cite

Monitoring antibiotic resistance genes in wastewater environments: The challenges of filling a gap in the One-Health cycle

Cited by 72 publications

References 114 publications

Metagenomic profiling and transfer dynamics of antibiotic resistance determinants in a full-scale granular sludge wastewater treatment plant

Metagenomic profiling and transfer dynamics of antibiotic resistance determinants in a full-scale granular sludge wastewater treatment plant

Development and validation of novel PCR primers for identification of plasmid-mediated colistin resistance (mcr) genes in various environmental settings

First detection of vanA positive Enterococcus faecium clonal complex 17 in hospital wastewater in Algeria: an epidemiological report

Contact Info

Product

Resources

About