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Given the rising prevalence of antibiotic-resistant\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      , phages are increasingly regarded as promising alternatives to conventional antibiotics. Henuyfy11N was isolated using the double-layer agar method and characterized via transmission electron microscopy in this study. Biological assays included stability under varying pH and temperature, UV sensitivity, host range, optimal multiplicity of infection, adsorption rate, and one-step growth curve.\n                      <jats:italic toggle=\"yes\">In vitro<\/jats:italic>\n                      lytic activity against extended-spectrum \u03b2-lactamase (ESBL)-producing\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      and biofilm eradication capacity was assessed. Whole-genome sequencing enabled phylogenetic, synteny (the analysis of conserved blocks of genetic sequence between different genomes), and functional annotation analyses.\n                      <jats:italic toggle=\"yes\">In vivo<\/jats:italic>\n                      , the therapeutic efficacy was evaluated in a mouse infection model. Phage Henuyfy11N has not yet been classified. It demonstrated high lytic activity, a short latent period, and a burst size of 57.1 PFU\/cell. The phage remained stable across a broad pH range (3\u201311) and temperatures up to 70\u2103. Its circular double-stranded DNA genome (41,103 bp, G + C% 50.88) contains 54 open reading frames, with no tRNA, virulence, or antibiotic resistance genes. Genomic and phylogenetic analyses revealed close relatedness to phage BUCT789. Henuyfy11N effectively lysed ESBL-producing\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      , disrupted biofilms, and significantly improved survival in the mouse infection model. Henuyfy11N shows high host specificity, efficient lytic activity, rapid replication, and a safe genomic profile, demonstrating some potential as a therapeutic agent against ESBL-producing\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      infections.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        The widespread use of antibiotics has led to increasing antibiotic resistance, which is a growing global health concern. Therefore, the development of novel antimicrobial therapy that can cure drug-resistant bacteria-induced infections is imperative. Phages are of increasing interest as natural enemies of bacteria, with clear advantages in antibacterial applications. In this study, by using extended-spectrum \u03b2-lactamase (ESBL)-producing\n                        <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                        2025011N as a host, we successfully isolated and purified\n                        <jats:italic toggle=\"yes\">Escherichia<\/jats:italic>\n                        phage Henuyfy11N and conducted a series of experiments to verify its genomic character and biological character. Our findings revealed that the phage exhibited excellent tolerance to a broad spectrum of pH and wide temperature range. Phage Henuyfy11N was effective in disrupting mature biofilm, and no genes for virulence, lysogenic, integrase, or AMRs were found in the genome. Besides, Henuyfy11N showed promising antibacterial effects\n                        <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                        , indicating potential as a therapeutic agent against ESBL-producing\n                        <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                        infections.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03253-25","type":"journal-article","created":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T09:06:03Z","timestamp":1777280763000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Characterization of novel phage Henuyfy11N: a potential therapeutic agent against extended-spectrum \u03b2-lactamase (ESBL)-producing\n                    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