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  • Eugenol as a Novel Antibiotic Adjuvant Against Carbapenem-Resistant Acinetobacter baumannii: Synergistic Activity With Carbapenems and Downregulation of OXA-51

  • Sheyda Sharabiani,1 Shirin Mohammadipour,2 Masoumeh Abed,3 Danial Ansari talebi,4 Pejman Rastar,5 Leila Asadpour,6,*
    1. Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran.
    2. Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran
    3. Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran.
    4. DVM, Faculty of Veterinary Medicine, Islamic Azad University, Shoushtar Branch, Shoushtar, Iran
    5. Department of Animal Sciences and Veterinary Medicine, Faculty of Agriculture, Water, Food and Functional Foods, Islamic Azad University, Rasht Branch, Rasht, Iran
    6. Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran.


  • Introduction: Carbapenem-resistant Acinetobacter baumannii is an important multidrug-resistant pathogen associated with limited therapeutic options. The increasing prevalence of carbapenem resistance highlights the need for novel compounds capable of restoring or enhancing antibiotic activity. Eugenol, a bioactive natural compound, has demonstrated antibacterial potential and may act synergistically with conventional antibiotics. This study investigated the antibacterial activity of eugenol alone and in combination with carbapenems against resistant A. baumannii isolates and evaluated its potential effect on the expression of the carbapenemase-associated gene OXA-51.
  • Methods: Ten clinical A. baumannii isolates were initially evaluated for antimicrobial susceptibility. Four isolates showing complete resistance to both imipenem and meropenem were selected for subsequent experiments. The antibacterial activity of eugenol was assessed by disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. Synergistic interactions between eugenol and imipenem or meropenem were evaluated using the checkerboard method and fractional inhibitory concentration index (FICI). The effect of sub-MIC eugenol on OXA-51 expression was assessed by real-time PCR in isolates carrying this gene.
  • Results: Eugenol exhibited clear antibacterial activity against all four selected carbapenem-resistant isolates, producing inhibition zones of 21–27 mm at a concentration of 1 mg. Eugenol MIC values were 250 µg/mL in two isolates and 500 µg/mL in the remaining two isolates, while the MBC was 500 µg/mL for all four isolates. Checkerboard analysis demonstrated a synergistic interaction between eugenol and imipenem in two isolates (FICI = 0.5), a partial synergistic interaction in one isolate (FICI = 0.75), and an additive interaction in one isolate (FICI = 1.0). The combination of eugenol with meropenem showed complete synergy in three isolates and partial synergy in one isolate. Molecular analysis further demonstrated that sub-MIC eugenol significantly reduced OXA-51 expression by at least 50% in the gene-positive isolates (p < 0.05).
  • Conclusion: Eugenol demonstrated direct antibacterial activity against carbapenem-resistant A. baumannii and enhanced the activity of both imipenem and meropenem through synergistic interactions. The substantial reduction in OXA-51 expression provides additional molecular evidence supporting its potential as an antibiotic-adjuvant strategy. These findings suggest that eugenol may represent a promising novel approach for improving the efficacy of carbapenems against resistant A. baumannii.
  • Keywords: Acinetobacter baumannii; carbapenem resistance; eugenol; antibiotic synergy; OXA-51

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