# Phenotypic and genotypic characterization of probiotic strains in the context of antimicrobial resistance

**Authors:** Ádám Kerek, Nikolett Palkovicsné Pézsa, Eszter Kaszab, Ákos Jerzsele, Orsolya Farkas

PMC · DOI: 10.3389/fvets.2025.1684650 · Frontiers in Veterinary Science · 2025-10-03

## TL;DR

This study examines the antibiotic resistance of probiotic bacteria, finding some strains carry resistance genes, which raises safety concerns for their use in humans and animals.

## Contribution

The study provides a combined phenotypic and genotypic analysis of antimicrobial resistance in industrially used probiotic strains.

## Key findings

- Multiple probiotic strains showed resistance to clinically relevant antibiotics like gentamicin and amoxicillin.
- Whole-genome sequencing identified 27 distinct antimicrobial resistance genes across the strains.
- Bacillus licheniformis had the most diverse resistance gene profile, while Pediococcus acidilactici had none.

## Abstract

Antimicrobial resistance (AMR) has emerged as a critical global public health concern, particularly with regard to microorganisms used as probiotics in both veterinary and human healthcare. The aim of this study was to characterize the phenotypic and genotypic resistance profiles of several industrially applied probiotic bacterial strains, with special emphasis on the presence of antimicrobial resistance genes (ARGs).

Five strains, Enterococcus faecium, Bacillus licheniformis, Bacillus subtilis, Lactobacillus rhamnosus, and Pediococcus acidilactici were analyzed. Minimum inhibitory concentration (MIC) testing revealed resistance in multiple strains to clinically relevant antibiotics such as gentamicin, amoxicillin, tylosin, and florfenicol.

Whole-genome sequencing identified 27 distinct ARGs, primarily associated with efflux pumps and target protection or modification mechanisms. The Bacillus licheniformis strain harbored the most diverse ARG profile, whereas no resistance genes were detected in Pediococcus acidilactici.

These findings highlight the necessity of integrating phenotypic and genotypic assessments for the safe application of probiotic strains in animals and potentially human use.

## Linked entities

- **Chemicals:** gentamicin (PubChem CID 3467), amoxicillin (PubChem CID 33613), tylosin (PubChem CID 5280440), florfenicol (PubChem CID 114811)
- **Species:** Enterococcus faecium (taxon 1352), Bacillus licheniformis (taxon 1402), Bacillus subtilis (taxon 1423), Pediococcus acidilactici (taxon 1254)

## Full-text entities

- **Chemicals:** florfenicol (MESH:C035534), ARG (-), gentamicin (MESH:D005839), amoxicillin (MESH:D000658), tylosin (MESH:D015645)
- **Species:** Homo sapiens (human, species) [taxon 9606], Pediococcus acidilactici (species) [taxon 1254], Enterococcus faecium (species) [taxon 1352], Lacticaseibacillus rhamnosus (species) [taxon 47715], Bacillus licheniformis (species) [taxon 1402], Bacillus subtilis (species) [taxon 1423]

## Full text

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## Figures

2 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12533545/full.md

## References

84 references — full list in the complete paper: https://tomesphere.com/paper/PMC12533545/full.md

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Source: https://tomesphere.com/paper/PMC12533545