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<p>Bacteriophages show promise for microbiome engineering, but studying their transmission dynamics in multimember communities and animal hosts is technically challenging. We therefore created ‘Phollow’, a live imaging-based approach for tracking phage replication and spread in situ with single-virion resolution. Following interbacterial phage transmission is achieved by marking virions with distinct fluorescent proteins during assembly in newly infected cells. In vitro cell virology studies revealed clouds of phage virions dispersing upon bacterial lysis, leading to rampant transmission. Combining Phollow with optically transparent zebrafish, we visualized phage outbreaks within the vertebrate gut. We observed that virions from a zebrafish-derived <i>Plesiomonas</i> strain, but not a human-derived <i>E. coli</i>, rapidly disseminate systemically to the liver and brain. Moreover, antibiotics triggered waves of interbacterial transmission and sudden shifts in gut community ecology. Phollow ultimately empowers multiscale investigations of phage transmission and transkingdom interactions that have the potential to open new avenues for phage-based microbiome therapies.</p>

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Phollow reveals in situ phage transmission dynamics in the zebrafish gut microbiome at single-virion resolution

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年4月18日

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  • Lizett Ortiz de Ora,
  • Elizabeth T. Wiles,
  • Mirjam Zünd,
  • Maria S. Bañuelos,
  • Nancy Haro-Ramirez,
  • Diana S. Suder,
  • Naveena Ujagar,
  • Julio Ayala-Angulo,
  • Calvin Trinh,
  • Courtney Knitter,
  • Shane Gonen,
  • Dequina A. Nicholas,
  • Travis J. Wiles

摘要

Bacteriophages show promise for microbiome engineering, but studying their transmission dynamics in multimember communities and animal hosts is technically challenging. We therefore created ‘Phollow’, a live imaging-based approach for tracking phage replication and spread in situ with single-virion resolution. Following interbacterial phage transmission is achieved by marking virions with distinct fluorescent proteins during assembly in newly infected cells. In vitro cell virology studies revealed clouds of phage virions dispersing upon bacterial lysis, leading to rampant transmission. Combining Phollow with optically transparent zebrafish, we visualized phage outbreaks within the vertebrate gut. We observed that virions from a zebrafish-derived Plesiomonas strain, but not a human-derived E. coli, rapidly disseminate systemically to the liver and brain. Moreover, antibiotics triggered waves of interbacterial transmission and sudden shifts in gut community ecology. Phollow ultimately empowers multiscale investigations of phage transmission and transkingdom interactions that have the potential to open new avenues for phage-based microbiome therapies.