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<p>The rise of methicillin-resistant <i>Staphylococcus pseudintermedius</i> (MRSP), a multidrug-resistant zoonotic pathogen with strong biofilm-forming ability, highlights the urgent need for alternative therapies. Here, we report the isolation and comprehensive characterization of vB_SpsS_VL4 (VL4), a novel temperate phage obtained from sewage using a clinical MRSP isolate. VL4 genome (39,788&#xa0;bp) contains 68 open reading frames (ORFs) with no virulence, resistance, or toxin genes and encodes lysis proteins with Cysteine, Histidine-dependent Amidohydrolase/Peptidase (CHAP) domains. Genomic and phylogenetic analyses classified VL4 as a new species within the <i>Coventryvirus</i> genus. Despite its temperate nature, VL4 demonstrated efficient replication (latent period 50&#xa0;min; burst size 56 PFU per cell), remained viable at neutral pH and temperatures up to 37&#xa0;°C, and significantly suppressed the host bacterial growth. Among 31 <i>S. pseudintermedius</i> clinical isolates, 27 (87%) strains were susceptible to VL4, while no activity was detected against <i>Staphylococcus aureus</i>. Moreover, VL4 inhibited biofilm formation and partially degraded mature MRSP biofilms. Collectively, these findings identify VL4 as a promising <i>Coventryvirus</i> phage with potential for future engineering and therapeutic development against difficult-to-treat MRSP infections.</p>

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Genomic and functional characterization of a novel temperate Coventryvirus phage infecting Staphylococcus pseudintermedius

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月22日

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  • Patoo Withatanung,
  • Sukanya Apiratwarrasakul,
  • Pathomporn Sresuwadjarey,
  • Nathita Phumthanakorn,
  • Varintip Lerdsittikul

摘要

The rise of methicillin-resistant Staphylococcus pseudintermedius (MRSP), a multidrug-resistant zoonotic pathogen with strong biofilm-forming ability, highlights the urgent need for alternative therapies. Here, we report the isolation and comprehensive characterization of vB_SpsS_VL4 (VL4), a novel temperate phage obtained from sewage using a clinical MRSP isolate. VL4 genome (39,788 bp) contains 68 open reading frames (ORFs) with no virulence, resistance, or toxin genes and encodes lysis proteins with Cysteine, Histidine-dependent Amidohydrolase/Peptidase (CHAP) domains. Genomic and phylogenetic analyses classified VL4 as a new species within the Coventryvirus genus. Despite its temperate nature, VL4 demonstrated efficient replication (latent period 50 min; burst size 56 PFU per cell), remained viable at neutral pH and temperatures up to 37 °C, and significantly suppressed the host bacterial growth. Among 31 S. pseudintermedius clinical isolates, 27 (87%) strains were susceptible to VL4, while no activity was detected against Staphylococcus aureus. Moreover, VL4 inhibited biofilm formation and partially degraded mature MRSP biofilms. Collectively, these findings identify VL4 as a promising Coventryvirus phage with potential for future engineering and therapeutic development against difficult-to-treat MRSP infections.