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<p>The rise of antimicrobial-resistant (AMR) pathogens poses a critical global public health threat. <i>Enterobacter cloacae</i> complex (ECC) causes severe, life-threatening infections due to both intrinsic and acquired resistance to last-line antibiotics. Its ability to form resilient biofilms further complicates treatment, underscoring the urgent need for innovative therapies. Phage therapy has emerged as a promising alternative, offering targeted antibacterial activity and the capacity to co-evolve with bacterial resistance. Here, we report the characterization of novel phages with potent antibiofilm activity against a multidrug-resistant <i>E. cloacae</i> clinical strain BOJ39. Phages were isolated from diverse environmental sources in Algeria. Host-range, thermal and pH stabilities, lytic and antibiofilm assays were assessed. Transmission electron microscopy (TEM) whole-genome sequencing with bioinformatic analyses were used for morphological and genomic characterization. Seventeen phages showing halo formation around plaques (designated ECHP) were selected. TEM revealed podovirus morphology. ECHP maintained infectivity across broad temperatures (2–70&#xa0;°C) and pH (2–12) ranges. Despite a narrow host range, ECHP showed strong lytic and antibiofilm activity against BOJ39. Genomic analysis revealed nine distinct genotypes among the ECHP isolates, which collectively constitute a putative novel species within the genus <i>Koutsourovirus</i>. No toxin, resistance, or virulence genes were detected, and a unique bifunctional virion-associated depolymerase was identified. ECHP Phages represent a novel <i>Koutsourovirus</i> lineage with strong lytic and anti-biofilm activity against multidrug-resistant and biofilm forming ECC. Their genetic safety and anti-biofilm potential highlight their promise as candidates for therapeutic development and as sources of novel antibacterial agents.</p>

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Novel Koutsourovirus phages disrupt clinical multidrug-resistant Enterobacter cloacae biofilms: genomic and functional characterization

  • 期刊论文

  • 发表日期：

    2026年1月26日

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  • Ibtihadj Souda,
  • Karima Boubekri,
  • Hibat Ellah Boutadjine,
  • Amal Thebti,
  • Hadda Imene Ouzari,
  • Benjamin D. Hatton,
  • Dalal Asker

摘要

The rise of antimicrobial-resistant (AMR) pathogens poses a critical global public health threat. Enterobacter cloacae complex (ECC) causes severe, life-threatening infections due to both intrinsic and acquired resistance to last-line antibiotics. Its ability to form resilient biofilms further complicates treatment, underscoring the urgent need for innovative therapies. Phage therapy has emerged as a promising alternative, offering targeted antibacterial activity and the capacity to co-evolve with bacterial resistance. Here, we report the characterization of novel phages with potent antibiofilm activity against a multidrug-resistant E. cloacae clinical strain BOJ39. Phages were isolated from diverse environmental sources in Algeria. Host-range, thermal and pH stabilities, lytic and antibiofilm assays were assessed. Transmission electron microscopy (TEM) whole-genome sequencing with bioinformatic analyses were used for morphological and genomic characterization. Seventeen phages showing halo formation around plaques (designated ECHP) were selected. TEM revealed podovirus morphology. ECHP maintained infectivity across broad temperatures (2–70 °C) and pH (2–12) ranges. Despite a narrow host range, ECHP showed strong lytic and antibiofilm activity against BOJ39. Genomic analysis revealed nine distinct genotypes among the ECHP isolates, which collectively constitute a putative novel species within the genus Koutsourovirus. No toxin, resistance, or virulence genes were detected, and a unique bifunctional virion-associated depolymerase was identified. ECHP Phages represent a novel Koutsourovirus lineage with strong lytic and anti-biofilm activity against multidrug-resistant and biofilm forming ECC. Their genetic safety and anti-biofilm potential highlight their promise as candidates for therapeutic development and as sources of novel antibacterial agents.