# Natural Essential Oils as Promising Antimicrobial Agents to Improve Food Safety: Mechanistic Insights Against Multidrug-Resistant Campylobacter jejuni and Campylobacter coli Isolated from Tunisia

## TL;DR

This study explores how natural essential oils can combat drug-resistant Campylobacter bacteria in food, offering a safer alternative to antibiotics.

## Contribution

The study identifies specific essential oils that effectively target multidrug-resistant Campylobacter species through membrane disruption.

## Key findings

- Eucalyptus globulus, Thymus algeriensis, and Myrtus communis showed strong inhibition of Campylobacter jejuni.
- Essential oils caused rapid bacterial reduction and membrane damage, as shown by time-kill and lysis experiments.
- EO treatments reduced salt tolerance and induced cytoplasmic leakage, confirming membrane disruption.

## Abstract

The increasing prevalence of multidrug-resistant (MDR) Campylobacter species poses a serious threat to food safety and public health, highlighting the urgent need for natural antimicrobial alternatives to conventional antibiotics. This study investigated the antibacterial potential and mechanism of action of seven essential oils (EOs), Cymbopogon citratus, Mentha pulegium, Artemisia absinthium, Myrtus communis, Thymus algeriensis, Thymus capitatus, and Eucalyptus globulus, against multidrug-resistant Campylobacter jejuni and Campylobacter coli. The antimicrobial activity was first assessed by the agar disk diffusion and broth microdilution methods to determine inhibition zones, minimum inhibitory concentrations (MICs), and minimum bactericidal concentrations (MBCs). The most active EOs were further evaluated through time–kill kinetics, cell lysis, salt tolerance, and membrane integrity assays to elucidate their bactericidal mechanisms. Results showed that E. globulus, T. algeriensis, and M. communis exhibited the strongest inhibitory effects, particularly against C. jejuni, with MIC values ranging from 3.125% to 6.25%, while C. coli was more resistant. Time–kill and lysis experiments demonstrated rapid bacterial reduction and significant decreases in optical density, indicating cell disruption. Additionally, EO treatments reduced salt tolerance and induced leakage of cytoplasmic materials, confirming membrane damage. Overall, these findings suggest that selected essential oils exert potent antimicrobial effects through membrane disruption and osmotic imbalance, offering promising natural strategies to control MDR Campylobacter in food systems. The application of such bioactive compounds could contribute significantly to improving food quality, extending shelf life, and enhancing food safety.