{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T20:07:09Z","timestamp":1791576429485,"version":"4.3.4"},"reference-count":46,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T00:00:00Z","timestamp":1740096000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004595","name":"Universiti Sains Malaysia","doi-asserted-by":"publisher","award":["R501-LR-RND003-0000000977-0000"],"award-info":[{"award-number":["R501-LR-RND003-0000000977-0000"]}],"id":[{"id":"10.13039\/501100004595","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Background<\/jats:title>\n<jats:p>The increasing incidence of AmpC \u03b2-lactamase producing by <jats:italic>K<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> has raised global alarm. Consequently, there is a crucial need for effective methods to inactivate pathogenic bacteria and mitigate the associated risks. Bacteriophage therapy has been demonstrated to be an effective and alternative approach for targeting and inactivating <jats:italic>K<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> that produces AmpC.<\/jats:p>\n<jats:p>This study aimed to isolate and characterize the <jats:italic>Klebsiella pneumoniae<\/jats:italic> AmpC-specific phages from hospital wastewater.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methods<\/jats:title>\n<jats:p>The hospital wastewater samples were collected from the sewage water effluent of a tertiary hospital at Universiti Sains Malaysia, located on the east coast of Malaysia. These samples underwent serial filtration and centrifugation processes for phage recovery. The phage solutions were undergoing a screening test by spot assay using clinical isolates of <jats:italic>Klebsiella pneumoniae<\/jats:italic> AmpC strain as amplification hosts. The isolated AmpC-phages were further studied and characterised to determine the phage\u2019s host range, temperature, pH, and chloroform stabilities. High-Resolution Transmission Electron Microscopy (HRTEM) was performed to determine the phage type.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Results<\/jats:title>\n<jats:p>Thirty HWW samples were analyzed using four K. pneumoniae AmpC strains resulting in a total of 120 screening plates. The AmpC\u2014Klebsiella pneumoniae (AmpC-KP) phages were detected in 31.70% (38\/120) of the plates. The AmpC-KP phages had lytic diameters ranging from 1\u20133 mm, and a phage titer ranged from4\u00d710<jats:sup>3<\/jats:sup>\u20133.2\u00d710<jats:sup>7<\/jats:sup> PFU\/ml. The phages had a narrow\u2013host range stable at a temperature range from -20\u201350\u02daC. The phages were also stable at pH ranging from 4 to 9 and at different concentrations of chloroform (5%,10%). Based on HRTEM, <jats:italic>Siphoviridea<\/jats:italic> was identified.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusions<\/jats:title>\n<jats:p>The AmpC-phages were abundant in hospital wastewater, and HWW was a good source for AmpC-KP phages. The isolated AmpC phages had a high effectivity and specificity for AmpC-KP with a narrow host range and could survive under harsh conditions such as (temperature, pH, and chloroform).<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0315079","type":"journal-article","created":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T18:22:34Z","timestamp":1740162154000},"page":"e0315079","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["Isolation and characterization of bacteriophage against clinical isolates of AmpC beta lactamase\u2013Producing Klebsiella pneumoniae from hospital wastewater"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8840-9139","authenticated-orcid":true,"given":"Manal Abdel Haleem A.","family":"Abusalah","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mai Abdel Haleem A.","family":"Abusalah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0034-7624","authenticated-orcid":true,"given":"Chan","family":"Yean Yean","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ismail","family":"Aziah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6612-8360","authenticated-orcid":true,"given":"Abdul Rahman","family":"Zaidah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,2,21]]},"reference":[{"issue":"8","key":"pone.0315079.ref001","doi-asserted-by":"crossref","first-page":"973","DOI":"10.14202\/vetworld.2017.973-978","article-title":"Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens","volume":"10","author":"RK Shende","year":"2017","journal-title":"Vet World"},{"issue":"1","key":"pone.0315079.ref002","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/s40560-020-0429-6","article-title":"Molecular diversity of extended-spectrum \u03b2-lactamases and carbapenemases, and antimicrobial resistance.","volume":"8","author":"T Sawa","year":"2020","journal-title":"Journal of Intensive Care"},{"issue":"3","key":"pone.0315079.ref003","article-title":"A Unified Numbering Scheme for Class C \u03b2-Lactamases","volume":"64","author":"MGP Page","year":"2020","journal-title":"Antimicrobial Agents and Chemotherapy"},{"key":"pone.0315079.ref004","doi-asserted-by":"crossref","first-page":"62","DOI":"10.12688\/aasopenres.13165.1","article-title":"Prevalence of plasmid-mediated AmpC beta-lactamases in Enterobacteria isolated from urban and rural folks in Uganda.","volume":"3","author":"CF Najjuka","year":"2020","journal-title":"AAS Open Res"},{"issue":"3","key":"pone.0315079.ref005","article-title":"Multidrug-Resistant ESBL\/AmpC-Producing Klebsiella pneumoniae Isolated from Healthy Thoroughbred Racehorses in Japan","volume":"10","author":"E Sukmawinata","year":"2020","journal-title":"Animals (Basel)."},{"issue":"3","key":"pone.0315079.ref006","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1016\/j.jcmgh.2022.11.005","article-title":"et al Klebsiella pneumoniae Induces Inflammatory Bowel Disease Through Caspase-11-Mediated IL18 in the Gut Epithelial Cells","volume":"15","author":"Q Zhang","year":"2023","journal-title":"Cell Mol Gastroenterol Hepatol"},{"issue":"1","key":"pone.0315079.ref007","first-page":"26","article-title":"et al Isolation and Characterization of Klebsiella Phages for Phage Therapy","volume":"2","author":"EM Townsend","year":"2021","journal-title":"Phage (New Rochelle)"},{"issue":"4","key":"pone.0315079.ref008","first-page":"277","article-title":"The antibiotic resistance crisis: part 1: causes and threats","volume":"40","author":"CL Ventola","year":"2015","journal-title":"P t"},{"issue":"6","key":"pone.0315079.ref009","doi-asserted-by":"crossref","first-page":"3417","DOI":"10.1128\/AEM.70.6.3417-3424.2004","article-title":"Evaluation of a cocktail of three bacteriophages for biocontrol of Escherichia coli O157:H7","volume":"70","author":"G O\u2019Flynn","year":"2004","journal-title":"Appl Environ Microbiol"},{"issue":"8","key":"pone.0315079.ref010","doi-asserted-by":"crossref","first-page":"2037","DOI":"10.1007\/s00705-018-3938-z","article-title":"Virus classification\u2014where do you draw the line","volume":"163","author":"P Simmonds","year":"2018","journal-title":"Arch Virol"},{"key":"pone.0315079.ref011","doi-asserted-by":"crossref","first-page":"635597","DOI":"10.3389\/fcimb.2021.635597","article-title":"Rekindling of a Masterful Precedent; 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