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This study isolated and phenotypically characterized bacteriophages targeting MDR and biofilm-forming UPEC from hospital sewage in Ethiopia. Twenty-three archived MDR UPEC isolates were screened for biofilm formation, and phages were characterized by host range, efficiency of plating, multiplicity of infection (MOI), adsorption kinetics, one-step growth, environmental stability, and biofilm biomass reduction. Eight isolates (34.8%) formed biofilms, including 3 strong, 1 moderate, and 4 weak biofilm formers. Two phages, \u03a6ET-3175 and \u03a6FETA-207, were isolated with titers of 1.35\u2009\u00d7\u200910\u2079 and 1.12\u2009\u00d7\u200910\u2079 PFU\/mL, respectively. \u03a6ET-3175 showed broader activity, with detectable activity against 6\/23 (26.1%) isolates, an optimal MOI of 0.1, &gt;\u200995% adsorption within 20\u00a0min, a latent period of approximately 30\u00a0min, and a burst size of 147\u2009\u00b1\u20094.6 PFU per infected cell. Both phages retained detectable activity across pH 4\u201310 and at temperatures up to 50\u00a0\u00b0C. \u03a6ET-3175 reduced biofilm biomass by 3.2\u201339.5% among four susceptible biofilm-forming isolates, but the paired difference was not statistically significant (Z\u2009=\u2009\u2212\u20091.826,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.068). These findings provide preliminary in-vitro phenotypic evidence and support further genomic, structural, and biological characterization.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-73906-9","type":"journal-article","created":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T09:15:31Z","timestamp":1790846131000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation and phenotypic characterization of bacteriophages targeting multidrug-resistant biofilm-forming uropathogenic Escherichia coli from hospital sewage"],"prefix":"10.1038","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-3102-6766","authenticated-orcid":false,"given":"Ephrem","family":"Tamrat","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6949-3494","authenticated-orcid":false,"given":"Awoke","family":"Derbie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2331-0557","authenticated-orcid":false,"given":"Feleke","family":"Mekonnen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-8076-0179","authenticated-orcid":false,"given":"Zelalem","family":"Asmare","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7153-4849","authenticated-orcid":false,"given":"Alene","family":"Geteneh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5259-2245","authenticated-orcid":false,"given":"Tesfaye","family":"Kassa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,10,1]]},"reference":[{"issue":"8","key":"73906_CR1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics9080466","volume":"9","author":"T De Miguel","year":"2020","unstructured":"De Miguel, T. et al. 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