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<p>The rising resistance of <i>Pseudomonas aeruginosa</i> to benzalkonium chloride (BAC) represents a major challenge in infection control. This study developed a benzalkonium chloride–essential oil nanoemulsion (BAC-EO-NE) using <i>Litsea cubeba</i> and <i>Melaleuca alternifolia</i> (tea tree) oils to overcome BAC resistance through terpene-mediated synergism. GC–MS profiling identified citronellal (17.3%), citronellol (13.8%), and 1,8-cineole (44.5%) as dominant membrane-active constituents. The optimized nanoemulsion displayed nanoscale droplet size (132.2&#xa0;nm), narrow PDI (0.162), and stable zeta potential (-13.5&#xa0;mV) with excellent thermodynamic stability. Biocompatibility studies revealed markedly reduced hemolysis (4.7 ± 0.21%) versus free BAC (9.2 ± 0.32%) and high fibroblast viability (&gt; 77%) after 72&#xa0;h. The BAC-EO-NE exhibited enhanced antibacterial activity, producing up to 2.7-fold larger inhibition zones and 4–sixfold lower MIC/MBC values compared to BAC. Notably, resistance in BAC-resistant <i>P. aeruginosa</i> was reversed, lowering MIC from &gt; 350 to 65&#xa0;µg/mL. The formulation effectively disrupted biofilms (MBEC 400–750&#xa0;ppm) and achieved superior contact-killing kinetics on treated textiles. SYTOX Green assays confirmed increased membrane permeability in <i>P. aeruginosa</i> and Methicillin-resistant <i>S. aureus</i> (MRSA). In an ex vivo porcine-skin disinfection model, BAC-EO-NE completely eradicated <i>S. aureus</i> within 96&#xa0;h, outperforming mupirocin. These findings demonstrate, for the first time, that plant volatile nanoemulsion-assisted membrane disruption enhances bacterial susceptibility to BAC and improves antibacterial activity. The BAC-EO-NE represents a promising, biocompatible strategy for next-generation disinfectants and topical antimicrobials targeting resistant pathogens.</p> Graphical Abstract <p></p>

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Plant Volatile Nanoemulsion Resensitize Pseudomonas aeruginosa to Benzalkonium Chloride

  • 期刊论文

  • 发表日期：

    2026年7月20日

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  • Ngurang Nisha,
  • Pompi Das,
  • Karpi Ango,
  • Debajit Mahanta,
  • Sanjeeb Kalita

摘要

The rising resistance of Pseudomonas aeruginosa to benzalkonium chloride (BAC) represents a major challenge in infection control. This study developed a benzalkonium chloride–essential oil nanoemulsion (BAC-EO-NE) using Litsea cubeba and Melaleuca alternifolia (tea tree) oils to overcome BAC resistance through terpene-mediated synergism. GC–MS profiling identified citronellal (17.3%), citronellol (13.8%), and 1,8-cineole (44.5%) as dominant membrane-active constituents. The optimized nanoemulsion displayed nanoscale droplet size (132.2 nm), narrow PDI (0.162), and stable zeta potential (-13.5 mV) with excellent thermodynamic stability. Biocompatibility studies revealed markedly reduced hemolysis (4.7 ± 0.21%) versus free BAC (9.2 ± 0.32%) and high fibroblast viability (> 77%) after 72 h. The BAC-EO-NE exhibited enhanced antibacterial activity, producing up to 2.7-fold larger inhibition zones and 4–sixfold lower MIC/MBC values compared to BAC. Notably, resistance in BAC-resistant P. aeruginosa was reversed, lowering MIC from > 350 to 65 µg/mL. The formulation effectively disrupted biofilms (MBEC 400–750 ppm) and achieved superior contact-killing kinetics on treated textiles. SYTOX Green assays confirmed increased membrane permeability in P. aeruginosa and Methicillin-resistant S. aureus (MRSA). In an ex vivo porcine-skin disinfection model, BAC-EO-NE completely eradicated S. aureus within 96 h, outperforming mupirocin. These findings demonstrate, for the first time, that plant volatile nanoemulsion-assisted membrane disruption enhances bacterial susceptibility to BAC and improves antibacterial activity. The BAC-EO-NE represents a promising, biocompatible strategy for next-generation disinfectants and topical antimicrobials targeting resistant pathogens.

Graphical Abstract