Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2
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 stud...
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| Format: | Article |
| Language: | English |
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BMC
2023-05-01
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| Series: | Virology Journal |
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| Online Access: | https://doi.org/10.1186/s12985-023-02034-x |
| _version_ | 1797806820518002688 |
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| author | Mohamed S. Fayez Toka A. Hakim Bishoy Maher Zaki Salsabil Makky Mohamed Abdelmoteleb Kareem Essam Anan Safwat Abdallah S. Abdelsattar Ayman El-Shibiny |
| author_facet | Mohamed S. Fayez Toka A. Hakim Bishoy Maher Zaki Salsabil Makky Mohamed Abdelmoteleb Kareem Essam Anan Safwat Abdallah S. Abdelsattar Ayman El-Shibiny |
| author_sort | Mohamed S. Fayez |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T06:13:10Z |
| format | Article |
| id | doaj.art-0681401a3c5c4ad08d54ccfaaf3d6481 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-03-13T06:13:10Z |
| publishDate | 2023-05-01 |
| publisher | BMC |
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| series | Virology Journal |
| spelling | doaj.art-0681401a3c5c4ad08d54ccfaaf3d64812023-06-11T11:05:52ZengBMCVirology Journal1743-422X2023-05-0120111610.1186/s12985-023-02034-xMorphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2Mohamed S. Fayez0Toka A. Hakim1Bishoy Maher Zaki2Salsabil Makky3Mohamed Abdelmoteleb4Kareem Essam5Anan Safwat6Abdallah S. Abdelsattar7Ayman El-Shibiny8Center for Microbiology and Phage Therapy, Zewail City of Science and TechnologyCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyDepartment of Botany, Faculty of Science, Mansoura UniversityCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyCenter for Microbiology and Phage Therapy, Zewail City of Science and TechnologyAbstract 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.https://doi.org/10.1186/s12985-023-02034-xGram negativeKlebsiella pneumoniaeSiphovirusMulti-drug resistance (MDR)BacteriophagePhage therapy |
| spellingShingle | Mohamed S. Fayez Toka A. Hakim Bishoy Maher Zaki Salsabil Makky Mohamed Abdelmoteleb Kareem Essam Anan Safwat Abdallah S. Abdelsattar Ayman El-Shibiny Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2 Virology Journal Gram negative Klebsiella pneumoniae Siphovirus Multi-drug resistance (MDR) Bacteriophage Phage therapy |
| title | Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2 |
| title_full | Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2 |
| title_fullStr | Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2 |
| title_full_unstemmed | Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2 |
| title_short | Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2 |
| title_sort | morphological biological and genomic characterization of klebsiella pneumoniae phage vb kpn zc2 |
| topic | Gram negative Klebsiella pneumoniae Siphovirus Multi-drug resistance (MDR) Bacteriophage Phage therapy |
| url | https://doi.org/10.1186/s12985-023-02034-x |
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