{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T05:28:37Z","timestamp":1790227717773,"version":"4.1.0"},"reference-count":49,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:00:00Z","timestamp":1759968000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Henry Ford Health Global Health Initiative"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p><jats:italic><jats:italic>Pseudomonas aeruginosa<\/jats:italic><\/jats:italic>, an opportunistic pathogen associated with wound infections, resists many commonly available antibiotics. Its ability to form biofilm provides an additional trait to evade antibiotics. Biofilm-associated infections are difficult to treat, raising the need for alternative strategies. Thus, this research aimed to investigate the potential of bacteriophage to disrupt the biofilm produced by <jats:italic><jats:italic>P. aeruginosa<\/jats:italic><\/jats:italic> isolated from wound infections. Wound samples were collected aseptically, processed for the isolation of <jats:italic><jats:italic>P. aeruginosa<\/jats:italic><\/jats:italic>, and identified by standard microbiological methods. Antimicrobial susceptibility was determined by the Kirby-Bauer disc diffusion method. Bacteriophages were isolated using the double-layer agar method. Phenotypic assessment of biofilm formation by the isolates and its reduction by phages was conducted by the tissue culture plate assay. Out of 647 wound samples processed, 96 <jats:italic><jats:italic>P. aeruginosa<\/jats:italic><\/jats:italic> were isolated<jats:italic>.<\/jats:italic> Piperacillin\/tazobactam was the most effective antibiotic, while doxycycline was the least effective. Among the total isolates, 86 (89.6%) were multidrug-resistant (MDR) and 69 (71.9%) were biofilm producers. Three different phages isolated from sewage demonstrated a high specificity to <jats:italic><jats:italic>P. aeruginosa<\/jats:italic><\/jats:italic>. Of these, phage vB_PaeP_PS2 lysed the highest number of isolates (22.9%), including 17 MDR and 21 biofilm-producing isolates. The biofilm reduction assay demonstrated that phage treatment significantly reduced biofilm formation, with vB_PaeP_PS2 achieving a 58% reduction after 6\u2009h of treatment. In conclusion, this study highlights the high prevalence of biofilm-producing MDR <jats:italic><jats:italic>P. aeruginosa<\/jats:italic><\/jats:italic> in wound infections and, for the first time in Nepal, demonstrates the potential of locally isolated phages to lyse biofilm-forming MDR isolates and disrupt their biofilms.<\/jats:p>","DOI":"10.1371\/journal.pone.0334139","type":"journal-article","created":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:34:48Z","timestamp":1760031288000},"page":"e0334139","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Bacteriophage as an anti-biofilm agent against Pseudomonas aeruginosa from wound infection"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-8260-022X","authenticated-orcid":true,"given":"Padma","family":"Shrestha","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-8116-5199","authenticated-orcid":true,"given":"Sujit","family":"Tandukar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-5474-6442","authenticated-orcid":true,"given":"Manisha","family":"Shrestha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanjit","family":"Shrestha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anup","family":"Subedee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9469-2148","authenticated-orcid":true,"given":"Jivan","family":"Shakya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9583-5374","authenticated-orcid":true,"given":"Reshma","family":"Tuladhar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,10,9]]},"reference":[{"issue":"10","key":"pone.0334139.ref001","doi-asserted-by":"crossref","first-page":"1162","DOI":"10.3390\/antibiotics10101162","article-title":"Microbial Species Isolated from Infected Wounds and Antimicrobial Resistance Analysis: Data Emerging from a Three-Years Retrospective Study","volume":"10","author":"V Puca","year":"2021","journal-title":"Antibiotics (Basel)"},{"issue":"7","key":"pone.0334139.ref002","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0179245","article-title":"Isolation and in vitro evaluation of bacteriophages against MDR-bacterial isolates from septic wound infections","volume":"12","author":"RR Pallavali","year":"2017","journal-title":"PLoS One"},{"key":"pone.0334139.ref003","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2017\/8953829","article-title":"Bacterial isolates and their antimicrobial susceptibility patterns of wound infections among inpatients and outpatients attending the University of Gondar Referral Hospital, Northwest Ethiopia","volume":"2017","author":"A Mohammed","year":"2017","journal-title":"Int J Microbiol"},{"issue":"7","key":"pone.0334139.ref004","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0270576","article-title":"Pseudomonas aeruginosa clinical blood isolates display significant phenotypic variability","volume":"17","author":"RJ Scheffler","year":"2022","journal-title":"PLoS One"},{"issue":"3","key":"pone.0334139.ref005","doi-asserted-by":"crossref","first-page":"100296","DOI":"10.1016\/j.infpip.2023.100296","article-title":"Multi-drug resistant Pseudomonas aeruginosa: a 2019-2020 single center retrospective case control study","volume":"5","author":"AF Yang","year":"2023","journal-title":"Infect Prev Pract"},{"key":"pone.0334139.ref006","volume-title":"WHO Bacterial Priority Pathogens List 2024 Bacterial Pathogens of Public Health Importance, to Guide Research, Development, and Strategies to Prevent and Control Antimicrobial Resistance","author":"WHO","year":"2024"},{"key":"pone.0334139.ref007","doi-asserted-by":"crossref","first-page":"1004733","DOI":"10.3389\/fmicb.2022.1004733","article-title":"Phage-resistant Pseudomonas aeruginosa against a novel lytic phage JJ01 exhibits hypersensitivity to colistin and reduces biofilm production","volume":"13","author":"W Wannasrichan","year":"2022","journal-title":"Front Microbiol"},{"key":"pone.0334139.ref008","doi-asserted-by":"crossref","first-page":"161","DOI":"10.2147\/NSA.S325594","article-title":"Bacterial Biofilm Destruction: A Focused Review On The Recent Use of Phage-Based Strategies With Other Antibiofilm Agents","volume":"14","author":"S Amankwah","year":"2021","journal-title":"Nanotechnol Sci Appl"},{"key":"pone.0334139.ref009","unstructured":"Sr S. 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