Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2

Virology Journal · 9 authors, 4 centres

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TL;DR

This study characterized a novel siphovirus bacteriophage (ZCKP2) isolated from sewage water, reporting a 48.2 kbp genome with no virulence, lysogeny, or antibiotic resistance genes, and demonstrating in vitro lytic activity against multidrug-resistant Klebsiella pneumoniae. Phage ZCKP2 showed stability across a range of temperatures and pH values and effective bacterial killing in a MOI-dependent manner.

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Phage vB_Kpn_ZC2 (ZCKP2) was isolated from sewage water using the clinical Klebsiella pneumoniae isolate KP/08 as host. ZCKP2 was identified as a siphovirus with a 48.2 kbp genome. The phage lysed 7 of 30 tested strains and displayed effective bacterial killing over 325 minutes in an MOI-dependent manner. Genomic analysis detected no lysogeny-related, antibiotic resistance, or virulence genes, supporting its safety profile. Genome annotation predicted adjacent genes encoding holin and lysozyme. These are in vitro findings; no in vivo efficacy or safety data are reported.

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other

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infectious disease

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ORIGINAL PAPER

PMC10158348

Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2

Virology Journal

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ABSTRACT

Abstract Background Bacteriophages (phages) are one of the most promising alternatives to traditional antibiotic therapies, especially against multidrug-resistant bacteria. Klebsiella pneumoniae is considered to be an opportunistic pathogen that can cause life-threatening infections. Thus, this study aims at the characterization of a novel isolated phage vB_Kpn_ZC2 (ZCKP2, for short). Methods The phage ZCKP2 was isolated from sewage water by using the clinical isolate KP/08 as a host strain. The isolated bacteriophage was purified and amplified, followed by testing of its molecular weight using Pulse-Field Gel Electrophoresis (PFGE), transmission electron microscopy, antibacterial activity against a panel of other Klebsiella pneumoniae hosts, stability studies, and whole genome sequencing. Results Phage ZCKP2 belongs morphologically to siphoviruses as indicated from the Transmission Electron Microscopy microgram. The Pulsed Field Gel Electrophoresis and the phage sequencing estimated the phage genome size of 48.2 kbp. Moreover, the absence of lysogeny-related genes, antibiotic resistance genes, and virulence genes in the annotated genome suggests that phage ZCKP2 is safe for therapeutic use. Genome-based taxonomic analysis indicates that phage ZCKP2 represents a new family that has not been formally rated yet. In addition, phage ZCKP2 preserved high stability at different temperatures and pH values (-20 − 70 °C and pH 4 – 9). For the antibacterial activity, phage ZCKP2 maintained consistent clear zones on KP/08 bacteria along with other hosts, in addition to effective bacterial killing over time at different MOIs (0.1, 1, and 10). Also, the genome annotation predicted antibacterial lytic enzymes. Furthermore, the topology of class II holins was predicted in some putative proteins with dual transmembrane domains that contribute significantly to antibacterial activity. Phage ZCKP2 characterization demonstrates safety and efficiency against multidrug-resistant K. pneumoniae , hence ZCKP2 is a good candidate for further in vivo and phage therapy clinical applications. Supplementary Information The online version contains supplementary material available at 10.1186/s12985-023-02034-x. Keywords: Gram negative, Klebsiella pneumoniae , Siphovirus, Multi-drug resistance (MDR), Bacteriophage, Phage therapy

CITATION EXPORT

Mohamed S Fayez, Toka A Hakim, Bishoy Maher Zaki et al.. Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2. Virology Journal. (2023). https://doi.org/10.1186/s12985-023-02034-x

AUTHORS

9 · 4 CENTRES

Mohamed S Fayez, Toka A Hakim, Bishoy Maher Zaki, Salsabil Makky, Mohamed Abdelmoteleb, Kareem Essam, Anan Safwat, Abdallah S Abdelsattar, Ayman El-Shibiny

1Center for Microbiology and Phage Therapy, Zewail City of Science and Technology, Giza, 12578 Egypt
2Microbiology and Immunology Department, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Giza, 11787 Egypt
3Department of Botany, Faculty of Science, Mansoura University, Mansoura, 35516 Egypt
4Faculty of Environmental Agricultural Sciences, Arish University, Arish, 45511 Egypt