{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:52:41Z","timestamp":1785909161680,"version":"3.56.0"},"reference-count":31,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      The increasing prevalence of carbapenem-resistant\n                      <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                      (CRKP) has posed a major challenge to clinical infection management. Phage therapy represents a promising alternative against multidrug-resistant bacterial infections; however, its application is limited by the scarcity of effective therapeutic phages. Therefore, the identification and characterization of novel phages are urgently needed.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      A novel Klebsiella pneumoniae phage was isolated and purified from hospital wastewater using the double-layer agar plate method with\n                      <jats:italic>K. pneumoniae<\/jats:italic>\n                      ATCC 700603 as the host strain. A total of 95 bacterial strains were used to evaluate the host range and lytic activity of the phage. The biological characteristics of the phage, including optimal multiplicity of infection (MOI), one-step growth curve, and stability under different environmental conditions (temperature, pH, ethanol, and ultraviolet exposure), were systematically investigated. Morphological characterization was performed by transmission electron microscopy. Whole-genome sequencing and bioinformatics analyses were conducted to determine genomic characteristics, taxonomic classification, and the presence of antibiotic resistance genes, virulence factors, or lysogeny-related genes to assess the therapeutic potential and biosafety of the phage.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The novel phage vB_KpnD_A2 exhibited potent lytic activity against multidrug-resistant Klebsiella pneumoniae strains. Host range analysis showed that vB_KpnD_A2 lysed 57.1% of carbapenem-resistant K. pneumoniae isolates and 63.6% of extended-spectrum \u03b2-lactamase-producing strains, while displaying specificity toward\n                      <jats:italic>K. pneumoniae<\/jats:italic>\n                      . The phage showed excellent biological properties, with an optimal MOI of 10\n                      <jats:sup>-<\/jats:sup>\n                      \u2076, a latent period of 40 min, and a burst size of 8.6\u00d710\u2074PFU\/cell. In addition, vB_KpnD_A2 maintained infectivity over a broad temperature range (4-70\u00b0C) and pH range (pH 2-13). Whole-genome analysis identified vB_KpnD_A2 as a strictly lytic Webervirus phage within the family Drexlerviridae, without detectable antibiotic resistance genes, virulence factors, or lysogeny-associated genes, supporting its therapeutic potential and biosafety.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>The newly characterized phage vB_KpnD_A2 exhibited potent lytic activity against selected multidrug-resistant Klebsiella pneumoniae isolates, favorable genomic safety, and good environmental stability. These findings support its potential as a therapeutic phage candidate for CRKP control and expand anti-Klebsiella phage resources for future phage-based interventions against antimicrobial-resistant bacterial infections.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1877260","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:41:19Z","timestamp":1785908479000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A novel bacteriophage capable of efficiently lysing multi-drug-resistant Klebsiella pneumoniae"],"prefix":"10.3389","volume":"17","author":[{"given":"Haipeng","family":"Zhang","sequence":"first","affiliation":[{"name":"Yunnan Provincial Key Laboratory of Natural Epidemic Diseases Prevention and Control Technology, Yunnan Institute of Endemic Diseases Prevention and Control","place":["Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qianwen","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Center for Global Health, School of Public Health, Nanjing Medical University","place":["Jiangsu, Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohong","family":"Nie","sequence":"additional","affiliation":[{"name":"The Laboratory Department of Dali Bai Autonomous Prefecture Chinese Medicine Hospital","place":["Yunnan, Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinlian","family":"Gao","sequence":"additional","affiliation":[{"name":"The Laboratory Department of Dali Bai Autonomous Prefecture Chinese Medicine Hospital","place":["Yunnan, Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunxian","family":"Yang","sequence":"additional","affiliation":[{"name":"The Laboratory Department of Dali Bai Autonomous Prefecture Chinese Medicine Hospital","place":["Yunnan, Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youhong","family":"Zhong","sequence":"additional","affiliation":[{"name":"Yunnan Provincial Key Laboratory of Natural Epidemic Diseases Prevention and Control Technology, Yunnan Institute of Endemic Diseases Prevention and Control","place":["Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liyuan","family":"Shi","sequence":"additional","affiliation":[{"name":"Yunnan Provincial Key Laboratory of Natural Epidemic Diseases Prevention and Control Technology, Yunnan Institute of Endemic Diseases Prevention and Control","place":["Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao","family":"Jin","sequence":"additional","affiliation":[{"name":"The Laboratory Department of Dali Bai Autonomous Prefecture Chinese Medicine Hospital","place":["Yunnan, Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Wang","sequence":"additional","affiliation":[{"name":"Yunnan Provincial Key Laboratory of Natural Epidemic Diseases Prevention and Control Technology, Yunnan Institute of Endemic Diseases Prevention and Control","place":["Dali, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1227147","DOI":"10.3389\/fmicb.2023.1227147","article-title":"Isolation and characterization of novel bacteriophage vB_KpP_HS106 for Klebsiella pneumonia K2 and applications in foods","volume":"14","author":"Chen","year":"2023","journal-title":"Front. Microbiol."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jviromet.2018.09.001","article-title":"Simple drop cast method for enumeration of bacteriophages","volume":"262","author":"Chhibber","year":"2018","journal-title":"J. Virol. Methods"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1186\/s40168-023-01547-5","article-title":"Water mass age structures the auxiliary metabolic gene content of free-living and particle-attached deep ocean viral communities","volume":"11","author":"Coutinho","year":"2023","journal-title":"Microbiome"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1186\/s12929-023-00946-y","article-title":"A novel phage carrying capsule depolymerase effectively relieves pneumonia caused by multidrug-resistant Klebsiella aerogenes","volume":"30","author":"Cui","year":"2023","journal-title":"J. Biomed. Sci."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"fiaf043","DOI":"10.1093\/femsec\/fiaf043","article-title":"Weberviruses are gut-associated phages that infect Klebsiella spp","volume":"101","author":"Dawson","year":"2025","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"103998","DOI":"10.1016\/j.ebiom.2022.103998","article-title":"Klebsiella species: taxonomy, hypervirulence and multidrug resistance","volume":"79","author":"Dong","year":"2022","journal-title":"EBioMedicine"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1265011","DOI":"10.3389\/fcimb.2023.1265011","article-title":"Isolation and characterization of three novel lytic phages against K54 serotype carbapenem-resistant hypervirulent Klebsiella pneumoniae","volume":"13","author":"Fang","year":"2023","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"4038","DOI":"10.3390\/ijms24044038","article-title":"Isolation and characterization of the first Zobellviridae family bacteriophage infecting Klebsiella pneumoniae","volume":"24","author":"Gorodnichev","year":"2023","journal-title":"Int. J. Mol. Sci."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"2450462","DOI":"10.1080\/21505594.2025.2450462","article-title":"Characterization and genomic insights into bacteriophages Kpph1 and Kpph9 against hypervirulent carbapenem-resistant Klebsiella pneumoniae","volume":"16","author":"Huang","year":"2025","journal-title":"Virulence"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"e0135324","DOI":"10.1128\/aem.01353-24","article-title":"Challenges and opportunities of phage therapy for Klebsiella pneumoniae infections","volume":"90","author":"Kou","year":"2024","journal-title":"Appl. Environ. Microbiol."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"301","DOI":"10.3390\/antibiotics9060301","article-title":"Growing trend of fighting infections in aquaculture environment-opportunities and challenges of phage therapy","volume":"9","author":"Kowalska","year":"2020","journal-title":"Antibiotics"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"792305","DOI":"10.3389\/fcimb.2021.792305","article-title":"Isolation and characterization of novel Phages targeting pathogenic Klebsiella pneumoniae","volume":"11","author":"Li","year":"2021","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"943279","DOI":"10.3389\/fmicb.2022.943279","article-title":"Biological and genomic characteristics of two bacteriophages isolated from sewage, using one multidrug-resistant and one non-multidrug-resistant strain of Klebsiella pneumoniae","volume":"13","author":"Liang","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1186\/s12866-022-02518-w","article-title":"Low-concentration iron promotes Klebsiella pneumoniae biofilm formation by suppressing succinic acid","volume":"22","author":"Liu","year":"2022","journal-title":"BMC Microbiol."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"1035364","DOI":"10.3389\/fcimb.2022.1035364","article-title":"Biological characteristics and genomic analysis of a novel Vibrio parahaemolyticus phage phiTY18 isolated from the coastal water of Xiamen China","volume":"12","author":"Liu","year":"2022","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"854908","DOI":"10.3389\/fmicb.2022.854908","article-title":"Isolation of Klebsiella pneumoniae phage vB_KpnS_MK54 and pathological assessment of endolysin in the treatment of pneumonia mice model","volume":"13","author":"Lu","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1186\/s12866-025-03813-y","article-title":"Characterization and genome analysis of novel Klebsiella pneumoniae phage vbKpUKJ_2 isolated from hospital sewage water","volume":"25","author":"Mirza","year":"2025","journal-title":"BMC Microbiol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"e0199422","DOI":"10.1128\/spectrum.01994-22","article-title":"Characterization of novel Klebsiella phage PG14 and its antibiofilm efficacy","volume":"10","author":"Mulani","year":"2022","journal-title":"Microbiol. Spectrum"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"1491961","DOI":"10.3389\/fmicb.2025.1491961","article-title":"Isolation, characterization, and genomic analysis of a novel bacteriophage vB_Kp_XP4 targeting hypervirulent and multidrug-resistant Klebsiella pneumoniae","volume":"16","author":"Peng","year":"2025","journal-title":"Front. Microbiol."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"62","DOI":"10.18502\/ijm.v15i1.11919","article-title":"Isolation of new Klebsiella pneumoniae phage PSKP16","volume":"15","author":"Sadeqi","year":"2023","journal-title":"Iran. J. Microbiol."},{"key":"ref21","volume-title":"Molecular Cloning: A Laboratory Manual","author":"Sambrook","year":"2001"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"85","DOI":"10.2174\/0109298665181166231212051621","article-title":"Molecular machinery of the triad Holin, endolysin, and Spanin: key players orchestrating bacteriophage-induced cell lysis and their therapeutic applications","volume":"31","author":"Samir","year":"2024","journal-title":"Protein Pept. Lett."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1186\/s12866-024-03355-9","article-title":"Analysis of a new phage, KZag1, infecting biofilm of Klebsiella pneumoniae: genome sequence and characterization","volume":"24","author":"Saqr","year":"2024","journal-title":"BMC Microbiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"1421724","DOI":"10.3389\/fcimb.2024.1421724","article-title":"Isolation and characterization of two novel bacteriophages against carbapenem-resistant Klebsiella pneumoniae","volume":"14","author":"Senhaji-Kacha","year":"2024","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1007\/s11262-024-02053-y","article-title":"Characterization and complete genome analysis of Klebsiella phage Kp109 with lytic activity against Klebsiella pneumoniae","volume":"60","author":"Singh","year":"2024","journal-title":"Virus Genes"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"lqab067","DOI":"10.1093\/nargab\/lqab067","article-title":"PHROG: families of prokaryotic virus proteins clustered using remote homology","volume":"3","author":"Terzian","year":"2021","journal-title":"NAR Genom. Bioinform."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"002111","DOI":"10.1099\/jgv.0.002111","article-title":"Summary of taxonomy changes ratified by the international committee on Taxonomy of viruses (ICTV) from the bacterial viruses subcommittee, 2025","volume":"106","author":"Turner","year":"2025","journal-title":"J. Gen. Virol."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1186\/s12866-024-03311-7","article-title":"Performance comparison of BD Phoenix CPO detect panel with cepheid Xpert Carba-R assay for the detection of carbapenemase-producing Klebsiella pneumoniae isolates","volume":"24","author":"T\u00fczemen","year":"2024","journal-title":"BMC Microbiol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"2339","DOI":"10.1007\/s00705-019-04309-7","article-title":"Characterization and complete genome sequence analysis of phage GP4, a novel lytic Bcep22-like podovirus","volume":"164","author":"Wang","year":"2019","journal-title":"Arch. Virol."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"3402","DOI":"10.13344\/j.microbiol.china.190367","article-title":"Characterization and genome analysis of a novel Klebsiella pneumoniae phage","volume":"46","author":"Wang","year":"2019","journal-title":"Microbiol. China"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"611","DOI":"10.3390\/v10110611","article-title":"Isolation of a novel Bacillus thuringiensis phage representing a new phage lineage and characterization of its endolysin","volume":"10","author":"Yuan","year":"2018","journal-title":"Viruses"}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1877260\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:41:20Z","timestamp":1785908480000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1877260\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":31,"alternative-id":["10.3389\/fmicb.2026.1877260"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2026.1877260","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]},"article-number":"1877260"}}