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Purpose <p>To investigate the efficacy of antimicrobial peptide CRAMP-34 against mono- and mixed-species biofilms formed by <i>Staphylococcus aureus</i> and <i>Pseudomonas aeruginosa</i>.</p> Method <p>By establishing an in vitro biofilm model, crystal violet staining and viable cell counting wereused to evaluate the eradication of the biofilm. Transcriptomics analysis combined with RT-qPCR wasemployed to elucidate the underlying mechanisms.</p> Result <p>CRAMP-34 significantly reduced biofilm biomass in both single-species and mixed speciesbiofilms across concentrations of 15.6–125 μg/mL, exhibiting a concentration-dependent effect. Usingselective agar plates, we demonstrated that CRAMP-34 significantly reduced the viable counts of both <i>P. aeruginosa</i> and <i>S. aureus</i> within mixed-species biofilms. Transcriptomic profiling of mature mixed species biofilms treated with 62.5 μg/mL CRAMP-34 revealed 86 differentially expressed biofilm-related genes in <i>P. aeruginosa</i>, predominantly involving exopolysaccharide biosynthesis and quorumsensing regulation. Subsequent assays confirmed altered alginate and pyocyanin expression, alongsideimpaired bacterial swimming motility, consistent with transcriptional changes. Furthermore, RT qPCRanalysis showed that CRAMP-34 downregulated key biofilm-associated genes in <i>S. aureus</i>, including <i>icaA</i> and <i>agrA</i>.</p> Conclusion <p>These findings indicate that CRAMP-34 disrupts <i>P. aeruginosa–S. aureus</i> biofilms byinterfering with exopolysaccharide production and Pseudomonas quinolone signal (PQS) systemregulation in <i>P. aeruginosa</i>, while also directly affecting S. aureus biofilm-related gene expression.Collectively, CRAMP-34 shows promising potential for preventing and treating biofilm-associatedinfections involving these two pathogens.</p>

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Eradication Efficacy of Antimicrobial Peptide CRAMP-34 Against Mixed-Species Biofilms Formed by Staphylococcus aureus and Pseudomonas aeruginosa

  • 期刊论文

  • 发表日期：

    2026年4月28日

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  • Huiming Hu,
  • Miao You,
  • Tengyue He,
  • Lihong Yuan,
  • Fan Wang,
  • Yanshuang Huang,
  • Huizhen Zhu,
  • Ruiqi Chen,
  • Shuting Liu,
  • Wei Wei,
  • Hongzao Yang,
  • Hongwei Chen

摘要

Purpose

To investigate the efficacy of antimicrobial peptide CRAMP-34 against mono- and mixed-species biofilms formed by Staphylococcus aureus and Pseudomonas aeruginosa.

Method

By establishing an in vitro biofilm model, crystal violet staining and viable cell counting wereused to evaluate the eradication of the biofilm. Transcriptomics analysis combined with RT-qPCR wasemployed to elucidate the underlying mechanisms.

Result

CRAMP-34 significantly reduced biofilm biomass in both single-species and mixed speciesbiofilms across concentrations of 15.6–125 μg/mL, exhibiting a concentration-dependent effect. Usingselective agar plates, we demonstrated that CRAMP-34 significantly reduced the viable counts of both P. aeruginosa and S. aureus within mixed-species biofilms. Transcriptomic profiling of mature mixed species biofilms treated with 62.5 μg/mL CRAMP-34 revealed 86 differentially expressed biofilm-related genes in P. aeruginosa, predominantly involving exopolysaccharide biosynthesis and quorumsensing regulation. Subsequent assays confirmed altered alginate and pyocyanin expression, alongsideimpaired bacterial swimming motility, consistent with transcriptional changes. Furthermore, RT qPCRanalysis showed that CRAMP-34 downregulated key biofilm-associated genes in S. aureus, including icaA and agrA.

Conclusion

These findings indicate that CRAMP-34 disrupts P. aeruginosa–S. aureus biofilms byinterfering with exopolysaccharide production and Pseudomonas quinolone signal (PQS) systemregulation in P. aeruginosa, while also directly affecting S. aureus biofilm-related gene expression.Collectively, CRAMP-34 shows promising potential for preventing and treating biofilm-associatedinfections involving these two pathogens.