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ORIGINAL RESEARCH article

Front. Microbiol.

Sec. Phage Biology

Genomic and Functional Characterization of a Lytic Klebsiella Phage UHKP with Antibiofilm Activity

Provisionally accepted
Muhammad HassanMuhammad Hassan1Iqbal Ahmad AlviIqbal Ahmad Alvi2,3*Sadiq Noor KhanSadiq Noor Khan1Dawood AhmedDawood Ahmed1*Muhammad AsifMuhammad Asif4Afshan SaleemAfshan Saleem5
  • 1Department of Medical Lab Technology, The University of Haripur, Haripur, Khyber Pakhtunkhwa, Pakistan, Haripur, Pakistan
  • 2Department of Microbiology, Afghan International Islamic University, Kabul, Afghanistan, Kabul, Afghanistan
  • 3Department of Microbiology, Hazara University Mansehra, Khyber Pakhtunkhwa, Pakistan, Mansehra, Pakistan
  • 4Department of Microbiology, Government College University Lahore, Punjab, Pakistan, lahore, Pakistan
  • 5Department of Microbiology, The University of Haripur, Haripur, Khyber Pakhtunkhwa, Pakistan, Haripur, Pakistan

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Klebsiella pneumoniae is an opportunistic pathogen causing severe hospital-acquired infections, and it rapidly acquires multidrug resistance. Its robust biofilm formation further complicates treatment and drives interest in phage therapy. A phage UHKP was isolated from hospital sewage using an MDR K. pneumoniae strain (KP-03). UHKP formed clear plaques and having phage titer 2.3×109 PFU/mL. Host-range testing on 19 clinical isolates showed a narrow spectrum. Only four MDR strains (KP-03, KP-05, KP-08, KP-11) and one K-17 serotype were lysed, with no activity on other strains or species. One-step growth analysis yielded a 30 min latent period and 85 PFU burst size. In planktonic culture, UHKP at MOI 1 stopped bacterial growth by 4 h and cleared cultures by 8 h, whereas at MOI 0.1 killing was delayed and incomplete. In static biofilm assays, UHKP eradicated 98% of 24-h and 96% of 48-h biofilm biomass by 24 h (MOI 1). Clearance of 72–96 h biofilms was limited (≤87% by 24 h). UHKP possesses an icosahedral head of 56 ± 3 nm and a short, non-contractile tail measuring around 15 ± 2 nm. Genome sequencing revealed a 62,542 bp dsDNA genome (56.6% GC) encoding 77 ORFs, and phylogenetic analysis placed UHKP in the genus Lastavirus. UHKP carries no lysogeny, toxin or antibiotic-resistance genes.

Keywords: Bacteriophage therapy, biofilm disruption, Genomic characterization, Klebsiella pneumoniae, multidrug resistance, UHKP

Received: 25 Dec 2025; Accepted: 03 Feb 2026.

Copyright: © 2026 Hassan, Alvi, Khan, Ahmed, Asif and Saleem. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Iqbal Ahmad Alvi
Dawood Ahmed

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Supplementary Material