# Comparative core-genome MLST of vancomycin-resistant Enterococcus faecium supports the utility of wastewater-based surveillance: a pilot study

**Authors:** Ahmad Ibrahim Al-Mustapha, Laura Lindholm, Riikka Laukkanen-Ninios, Venla Johansson, Ananda Tiwari, Viivi Heljanko, Kirsi-Maarit Lehto, Anssi Lipponen, Sami Oikarinen, Tarja Pitkänen, Annamari Heikinheimo, Anna-Maria Hokajärvi, Anna-Maria Hokajärvi, Anniina Sarekoski, Aleksi Kolehmainen, Soile Blomqvist, Kati Räisänen, Carita Savolainen-Kopra, Teemu Möttönen, Oskari Luomala, Aapo Juutinen, Annika Länsivaara, Rafiqul Hyder, Erja Janhonen, Paula Kurittu

PMC · DOI: 10.1186/s12866-025-04610-3 · BMC Microbiology · 2026-01-05

## TL;DR

This study shows that wastewater can be used to track antibiotic-resistant bacteria, like vancomycin-resistant Enterococcus faecium, and aligns with human surveillance data.

## Contribution

The study demonstrates the utility of wastewater-based surveillance for tracking AMR in VREfm using core-genome MLST.

## Key findings

- VREfm was isolated from 22% of wastewater samples, with no significant variation across treatment plants.
- Most wastewater isolates clustered closely with human isolates, showing genetic similarity.
- Wastewater-based surveillance aligns with national human surveillance data for VREfm.

## Abstract

Wastewater-based surveillance (WBS) could complement clinical data and be used as an early warning tool for population-level monitoring of priority pathogens such as vancomycin-resistant Enterococcus faecium (VREfm). In this pilot study, we isolated VREfm using CHROMAgar VRE from 77 composite wastewater influent samples from ten wastewater treatment plants (WWTPs), assessed their phenotypic antibiotic susceptibility profiles using the broth microdilution assay, and used core genome MLST (cgMLST) to examine the genetic relatedness of human and wastewater isolates in Finland. VREfm was isolated from 17 samples (22%), with no significant difference in the isolation rate across the ten WWTPs (p = 0.407). The phenotypic antimicrobial susceptibility testing (AST) revealed that all isolates were resistant to ampicillin, ciprofloxacin, erythromycin, teicoplanin, and vancomycin and were susceptible to chloramphenicol, daptomycin, and linezolid. All 17 VREfm isolates belonged to CC17, and 12 of them were sequence type ST80. Altogether, 76% (n = 13) of the wastewater isolates harboured the vanA gene, 18% (n = 3) had the vanB gene, and 6% (n = 1) had both vanA and vanB genes. cgMLST revealed that most VREfm strains from wastewater clustered closely (< 10 allelic differences) with those from human surveillance isolates across the country. The ST80 isolates belonged to the clusters identified during 2022–2023 as part of national VRE surveillance of screening samples. Therefore, WBS closely aligns with what is observed through human surveillance. We believe that this pilot supports the utility of WBS as a sensitive tool for AMR surveillance. This study also contributes to the One Health approach, needed to fully understand the transmission dynamics of pathogenic bacteria and effectively manage the antimicrobial resistance (AMR) challenge.

The online version contains supplementary material available at 10.1186/s12866-025-04610-3.

## Linked entities

- **Genes:** vanA (vanillate O-demethylase oxygenase) [NCBI Gene 877879], vanB (vanillate O-demethylase) [NCBI Gene 877880]
- **Chemicals:** ampicillin (PubChem CID 6249), ciprofloxacin (PubChem CID 2764), erythromycin (PubChem CID 12560), teicoplanin (PubChem CID 133065662), vancomycin (PubChem CID 14969), chloramphenicol (PubChem CID 5959), daptomycin (PubChem CID 21585658), linezolid (PubChem CID 3929)
- **Species:** Enterococcus faecium (taxon 1352)

## Full-text entities

- **Chemicals:** ampicillin (MESH:D000667), erythromycin (MESH:D004917), ciprofloxacin (MESH:D002939), teicoplanin (MESH:D017334), vancomycin (MESH:D014640), daptomycin (MESH:D017576), linezolid (MESH:D000069349), chloramphenicol (MESH:D002701)
- **Species:** Homo sapiens (human, species) [taxon 9606], Enterococcus faecium (species) [taxon 1352]

## Full text

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## Figures

2 figures with captions in the complete paper: https://tomesphere.com/paper/PMC13005440/full.md

## References

4 references — full list in the complete paper: https://tomesphere.com/paper/PMC13005440/full.md

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Source: https://tomesphere.com/paper/PMC13005440