ORIGINAL RESEARCH
Isolation and characterization of Klebsiella pneumoniae bacteriophages encoding polysaccharide depolymerases with rare capsule specificity
1 Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia
2 Pavlov First Saint Petersburg State Medical University, Saint Petersburg, Russia
3 Pediatric Research and Clinical Center of Infectious Diseases of the Federal Medical Biological Agency, Saint Petersburg, Russia
Correspondence should be addressed: Roman B. Gorodnichev
Malaya Pirogovskaya, 1а, Moscow, 119435, Russia; moc.liamg@b.r.vehcindorog
Funding: the study was supported by the funds of the State Assignment "Development of the Scheme for Complex Therapy of Infectious Diseases Caused by Antibiotic Resistant Pathogens Involving the Use of Bacteriophages or Their Derivatives in Combination with Antibacterials” (code: Bacteriophage-2). Typing of Klebsiella pneumoniae strains was supported by the Russian Science Foundation (project No. 22-15-00149, https://rscf.ru/project/22-15-00149/).
Author contribution: Gorodnichev RB, Kornienko MA — study plan, data acquisition and processing, manuscript writing; Bespiatykh DA — data processing; Malakhova MV — data acquisition; Veselovsky VA, Goloshchapov OV, Chukhlovin AB, Bespyatykh JA — data acquisition and processing, Shitikov EA — study plan, data processing, manuscript writing.
Compliance with ethical standards: experimental work was carried out in compliance with the guidelines SP 1.3.2322-08 "Safety of Working With Microorganisms of III—IV Groups of Pathogenicity (Danger) and Causative Agents of Parasitic Diseases"; guidelines SP 1.3.2518-09 “Additions and Amendments № 1 to the guidelines SP 1.3.2322-08 "Safety of Working With Microorganisms of III—IV Groups of Pathogenicity (Danger) and Causative Agents of Parasitic Diseases"; guidelines "Sanitary and Epidemiologic Requirements for the Handling of Medical Waste" (SanPiN 2.1.7.2790-10 SanPiN 3.3686-21, SanPiN 2.1.3684-21); Federal Clinical Guidelines "Rational Use of Bacteriophages in Clinical and Epidemiological Practice".
Bacterial infections caused by antibiotic resistant strains of Klebsiella pneumoniae are among the most dangerous threats for the world's public healthcare. Treatment with bacteriophages and/or their derivatives could become one of the alternative methods for therapy of infections caused by K. pneumoniae. The study was aimed to isolate from the environment and characterize the capsule-specific K. pneumoniae bacteriophages that are useful for therapy and possess the polysaccharide depolymerase genes. Bacteriophages were isolated from the river water samples by enrichment method. The host range of bacteriophages were assessed using the collection of 180 K. pneumoniae clinical strains. Bacteriophage whole genome sequencing was performed on the MiSeq platform (Illumina). Four new bacteriophages from different taxonomic groups were isolated and characterized during the study: vB_KpnM_NDO71 (Vequintavirinae family), vB_KpnS_MAG26fr (Casjensviridae family), vB_KpnS_MDA2066 (Ackermannviridae family), and vB_KpnS_PMM-G3 (Drexlerviridae family). Bacteriophages vB_KpnM_NDO71, vB_KpnS_MAG26fr, and vB_KpnS_PMM-G3 had a narrow lytic spectrum and lysed all strains with the capsular type of the host: KL45, KL19 or KL28, respectively. Bacteriophage vB_KpnS_MDA2066 showed lytic activity against strains with two different capsular types: KL19 and KL107. Bacteriophages were strictly virulent and contained no integrase genes, potentially dangerous toxin genes or antibiotic resistance determinants. This allows them to be used in therapeutic practice. Receptor-binding proteins represented by polysaccharide depolymerases were predicted for each bacteriophage.