{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T02:46:46Z","timestamp":1791946006729,"version":"4.3.4"},"reference-count":103,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T00:00:00Z","timestamp":1754438400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"DoD Congressionally Directed Medical Research Programs, Peer-Reviewed Medical Research Program, Focused Program Award","award":["PR182667"],"award-info":[{"award-number":["PR182667"]}]},{"name":"DoD Congressionally Directed Medical Research Programs, Peer-Reviewed Medical Research Program, Focused Program Award","award":["MI210045"],"award-info":[{"award-number":["MI210045"]}]},{"name":"Military Infectious Diseases Research Program","award":["PR182667"],"award-info":[{"award-number":["PR182667"]}]},{"name":"Military Infectious Diseases Research Program","award":["MI210045"],"award-info":[{"award-number":["MI210045"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Multidrug-resistant (MDR) strains of Klebsiella pneumoniae present an acute threat as they continue to disseminate globally. Phage therapy has shown promise as a powerful approach to combat MDR infections, but narrow phage host ranges make development of broad acting therapeutics more challenging. The goal of this effort was to use in vitro directed evolution (the \u201cAppelmans protocol\u201d) to isolate K. pneumoniae phages with broader host ranges for improved therapeutic cocktails. Five myophages in the genus Jiaodavirus (family Straboviridae) with complementary activity were mixed and passaged against a panel of 11 bacterial strains including a permissive host and phage-resistant clinical isolates. Following multiple rounds of training, we collected phage variants displaying altered specificity or expanded host ranges compared with parental phages when tested against a 100 strain diversity panel of K. pneumoniae. Some phage variants gained the ability to lyse previously phage-resistant strains but lost activity towards previously phage-susceptible strains, while several variants had expanded activity. Whole-genome sequencing identified mutations and recombination events impacting genes associated with host tropism including tail fiber genes that most likely underlie the observed changes in host ranges. Evolved phages with broader activity are promising candidates for improved K. pneumoniae therapeutic phage cocktails.<\/jats:p>","DOI":"10.3390\/ijms26157597","type":"journal-article","created":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T10:13:51Z","timestamp":1754475231000},"page":"7597","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Phage Host Range Expansion Through Directed Evolution on Highly Phage-Resistant Strains of Klebsiella pneumoniae"],"prefix":"10.3390","volume":"26","author":[{"given":"Kevin A.","family":"Burke","sequence":"first","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4644-8743","authenticated-orcid":false,"given":"Tracey L.","family":"Peters","sequence":"additional","affiliation":[{"name":"Department of Entomology, College of Agricultural, Human and Natural Resource Sciences, Washington State University, Pullman, WA 99164, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga A.","family":"Kirillina","sequence":"additional","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-7696-1808","authenticated-orcid":false,"given":"Caitlin D.","family":"Urick","sequence":"additional","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bertran D.","family":"Walton","sequence":"additional","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5753-6058","authenticated-orcid":false,"given":"Jordan T.","family":"Bird","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nino","family":"Mzhavia","sequence":"additional","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin O.","family":"Georges","sequence":"additional","affiliation":[{"name":"Department of Emerging Infectious Diseases, Walter Reed Army Institute of Research-Africa, Nairobi P.O. Box 606-00621, Kenya"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2993-9794","authenticated-orcid":false,"given":"Paphavee","family":"Lertsethtakarn","sequence":"additional","affiliation":[{"name":"Department of Bacterial and Parasitic Diseases, Walter Reed Army Institute of Research-Armed Forces Research Institute of Medical Sciences, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1418-6523","authenticated-orcid":false,"given":"Lillian A.","family":"Musila","sequence":"additional","affiliation":[{"name":"Department of Emerging Infectious Diseases, Walter Reed Army Institute of Research-Africa, Nairobi P.O. Box 606-00621, Kenya"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5770-8980","authenticated-orcid":false,"given":"Mikeljon P.","family":"Nikolich","sequence":"additional","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2256-861X","authenticated-orcid":false,"given":"Andrey A.","family":"Filippov","sequence":"additional","affiliation":[{"name":"Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Chang, D., Sharma, L., Dela Cruz, C.S., and Zhang, D. 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