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<p>The increasing prevalence of extended-spectrum β-lactamase (ESBL)-producing uropathogenic <i>Escherichia coli</i> (UPEC), particularly strains with multidrug resistance (MDR) and enhanced biofilm-forming capacity, represents a major challenge in the management of urinary tract infections. This study investigated the antibacterial, antibiofilm, and mechanistic effects of 1,8-cineole against MDR ESBL-producing UPEC isolates. Antibacterial activity was evaluated using disk diffusion and minimum inhibitory concentration (MIC) assays. Membrane integrity, antibiofilm activity, and structural alterations were assessed through intracellular protein and nucleic acid leakage analyses, biofilm eradication assays, and scanning electron microscopy (SEM), respectively. Quantitative real-time PCR (qRT-PCR) was used to analyze the expression of biofilm-associated genes. Molecular docking was performed to investigate interactions between 1,8-cineole and major UPEC biofilm-related proteins. 1,8-Cineole exhibited marked antibacterial activity against MDR ESBL-producing UPEC isolates, with inhibition zone diameters of 13.4 ± 1.7&#xa0;mm and MIC values of 1.4 to 2.8&#xa0;mg/mL. Treatment induced significant leakage of intracellular proteins and nucleic acids, indicating disruption of membrane integrity. Exposure to 4 × MIC markedly impaired mature biofilm architecture. SEM analysis confirmed severe morphological damage, including membrane disruption and cellular deformation. qRT-PCR analysis demonstrated significant downregulation of the biofilm-associated genes <i>csgA</i>, <i>csgD</i>, <i>fimH</i>, <i>luxS</i>, and <i>pgaC</i> following treatment with 1,8-cineole. Molecular docking further revealed stable interactions between 1,8-cineole and key biofilm regulatory proteins, particularly with FimH. 1,8-Cineole exerts potent antibacterial and antibiofilm activity against MDR ESBL-producing UPEC through membrane destabilization and modulation of biofilm-associated pathways, supporting its potential as a plant-derived antimicrobial candidate for biofilm-associated urinary tract infections.</p>

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Exploring the antibacterial and antibiofilm potential of 1,8-cineole against Escherichia coli through in vitro, gene expression, and in silico analyses

  • 期刊论文

  • 发表日期：

    2026年8月4日

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  • Milad Kashi,
  • Amin Khoshbayan,
  • Zahra Chegini,
  • Aref Shariati

摘要

The increasing prevalence of extended-spectrum β-lactamase (ESBL)-producing uropathogenic Escherichia coli (UPEC), particularly strains with multidrug resistance (MDR) and enhanced biofilm-forming capacity, represents a major challenge in the management of urinary tract infections. This study investigated the antibacterial, antibiofilm, and mechanistic effects of 1,8-cineole against MDR ESBL-producing UPEC isolates. Antibacterial activity was evaluated using disk diffusion and minimum inhibitory concentration (MIC) assays. Membrane integrity, antibiofilm activity, and structural alterations were assessed through intracellular protein and nucleic acid leakage analyses, biofilm eradication assays, and scanning electron microscopy (SEM), respectively. Quantitative real-time PCR (qRT-PCR) was used to analyze the expression of biofilm-associated genes. Molecular docking was performed to investigate interactions between 1,8-cineole and major UPEC biofilm-related proteins. 1,8-Cineole exhibited marked antibacterial activity against MDR ESBL-producing UPEC isolates, with inhibition zone diameters of 13.4 ± 1.7 mm and MIC values of 1.4 to 2.8 mg/mL. Treatment induced significant leakage of intracellular proteins and nucleic acids, indicating disruption of membrane integrity. Exposure to 4 × MIC markedly impaired mature biofilm architecture. SEM analysis confirmed severe morphological damage, including membrane disruption and cellular deformation. qRT-PCR analysis demonstrated significant downregulation of the biofilm-associated genes csgA, csgD, fimH, luxS, and pgaC following treatment with 1,8-cineole. Molecular docking further revealed stable interactions between 1,8-cineole and key biofilm regulatory proteins, particularly with FimH. 1,8-Cineole exerts potent antibacterial and antibiofilm activity against MDR ESBL-producing UPEC through membrane destabilization and modulation of biofilm-associated pathways, supporting its potential as a plant-derived antimicrobial candidate for biofilm-associated urinary tract infections.