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<p>A combination approach using phytocompounds with antibiotics offers a promising strategy to enhance antibiotic efficacy and reduce pathogen virulence. This study investigates the synergistic antibacterial and anti-virulence potential of <i>Acacia nilotica</i> ethyl acetate fraction (ANEF) and its bioactive compound, methyl gallate (MG), in combination with meropenem against <i>Pseudomonas aeruginosa</i> PAO1. The synergy was assessed using checkerboard assays against <i>P. aeruginosa</i> and <i>Escherichia coli</i>, with ANEF-meropenem and MG-meropenem combinations yielding fractional inhibitory concentration index (FICI) values of 0.187 and 0.312, respectively, indicating strong synergism. These combinations significantly reduced bacterial viability, with log₁₀ CFU/ml reductions of 4.48 (ANEF-meropenem) and 3.68 (MG-meropenem). The combinations inhibited quorum sensing (QS)-regulated virulence factors, including pyocyanin, pyoverdine, protease, swarming, and EPS production by 58–82% at ½ FIC. The biofilm disruption assays further confirmed the anti-biofilm potential, with reductions of 86.67% by ANEF-meropenem and 82.66% for the MG-meropenem combination. The mechanistic evaluations revealed enhanced bacterial membrane disruption, with MG-meropenem showing the highest membrane permeability (54.63% dye uptake) and cellular leakage (OD 1.28). Hemolysis assays indicated dose-dependent toxicity, with maximum hemolysis by 45.39% (ANEF) and 29.94% (MG) at the MIC value. However, ANEF exhibited no adverse developmental effects in <i>Caenorhabditis elegans</i> N2 worms. These findings demonstrate that ANEF and MG, in synergy with meropenem, effectively impair <i>P. aeruginosa</i> growth, virulence, and biofilm formation while maintaining low toxicity profiles. The findings support the potential of phytochemical-antibiotic combinations as promising adjunct therapies for multidrug-resistant <i>P. aeruginosa</i> infections.</p>

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Anti-infective synergy of Acacia nilotica bioactive fraction and methyl gallate with meropenem against pathogenicity of Pseudomonas aeruginosa

  • 期刊论文

  • 发表日期：

    2025年6月11日

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  • Samreen,
  • Iqbal Ahmad

摘要

A combination approach using phytocompounds with antibiotics offers a promising strategy to enhance antibiotic efficacy and reduce pathogen virulence. This study investigates the synergistic antibacterial and anti-virulence potential of Acacia nilotica ethyl acetate fraction (ANEF) and its bioactive compound, methyl gallate (MG), in combination with meropenem against Pseudomonas aeruginosa PAO1. The synergy was assessed using checkerboard assays against P. aeruginosa and Escherichia coli, with ANEF-meropenem and MG-meropenem combinations yielding fractional inhibitory concentration index (FICI) values of 0.187 and 0.312, respectively, indicating strong synergism. These combinations significantly reduced bacterial viability, with log₁₀ CFU/ml reductions of 4.48 (ANEF-meropenem) and 3.68 (MG-meropenem). The combinations inhibited quorum sensing (QS)-regulated virulence factors, including pyocyanin, pyoverdine, protease, swarming, and EPS production by 58–82% at ½ FIC. The biofilm disruption assays further confirmed the anti-biofilm potential, with reductions of 86.67% by ANEF-meropenem and 82.66% for the MG-meropenem combination. The mechanistic evaluations revealed enhanced bacterial membrane disruption, with MG-meropenem showing the highest membrane permeability (54.63% dye uptake) and cellular leakage (OD 1.28). Hemolysis assays indicated dose-dependent toxicity, with maximum hemolysis by 45.39% (ANEF) and 29.94% (MG) at the MIC value. However, ANEF exhibited no adverse developmental effects in Caenorhabditis elegans N2 worms. These findings demonstrate that ANEF and MG, in synergy with meropenem, effectively impair P. aeruginosa growth, virulence, and biofilm formation while maintaining low toxicity profiles. The findings support the potential of phytochemical-antibiotic combinations as promising adjunct therapies for multidrug-resistant P. aeruginosa infections.