Klebsiella pneumoniae (family Enterobacteriaceae) is a gram-negative bacterium that has strong pathogenicity to humans and can cause sepsis, pneumonia, and urinary tract infection. In recent years, the unreasonable use of antibacterial drugs has led to an increase in drug-resistant strains of K. pneumoniae, a serious threat to public health. Bacteriophages, viruses that infect bacteria, are ubiquitous in the natural environment. They are considered to be the most promising substitute for antibiotics because of their high specificity, high efficiency, high safety, low cost, and short development cycle. In this study, a novel phage designated vB_KpnP_IME279 was successfully isolated from hospital sewage using a multidrug-resistant strain of K. pneumoniae as an indicator. A one-step growth curve showed that vB_KpnP_IME279 has a burst size of 140 plaque-forming units/cell and a latent period of 20 min at its optimal multiplicity of infection (MOI = 0.1). Phage vB_KpnP_IME279 survives in a wide pH range between 3 and 11 and is stable at temperatures ranging from 40 to 60 °C. Ten of the 20 strains of K. pneumoniae including the host bacteria were lysed by the phage vB_KpnP_IME279, and the multilocus sequence typing and wzi typing of the 10 strains were ST11, ST37, ST375, wzi209, wzi52, and wzi72, respectively. The genome of vB_KpnP_IME279 is 42,518 bp long with a G + C content of 59.3%. Electron microscopic observation showed that the phage belongs to the family Podoviridae. BLASTN alignment showed that the genome of the phage has low similarity with currently known phages. The evolutionary relationship between phage vB_KpnP_IME279 and other Podoviridae was analyzed using a phylogenetic tree based on sequences of phage major capsid protein and indicates that the phage vB_KpnP_IME279 belongs to the Podoviridae subfamily. These data enhance understanding of K. pneumoniae phages and will help in development of treatments for multidrug-resistant bacteria using phages.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriofágy klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• fylogeneze MeSH
• genom virový MeSH
• hostitelská specificita MeSH
• Klebsiella pneumoniae účinky léků   izolace a purifikace   virologie   MeSH
• lidé MeSH
• mikrobiologické techniky MeSH
• mnohočetná bakteriální léková rezistence MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• odpadní vody mikrobiologie   virologie   MeSH
• Podoviridae klasifikace   genetika   izolace a purifikace   MeSH
• RNA ribozomální 16S MeSH
• sekvenování celého genomu MeSH
• techniky typizace bakterií MeSH
• teplota MeSH
• zastoupení bazí MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Five bacteriophages (Kpn5, Kpn12, Kpn13, Kpn17 and Kpn22), each having specificity against Klebsiella pneumoniae strain B5055, were isolated from sewage samples and characterized in terms of growth characteristics, genetic material, morphology and structural proteins. Adsorption rate as well as single step growth curve experiments showed variation among phages. Restriction enzyme digestion of DNA confirmed the presence of double stranded DNA as well as the heterogeneous nature of genetic material. RAPD-PCR was performed to further distinguish these closely related phages. Their genome fingerprint confirmed their diversity. Transmission electron microscopy, on the other hand, showed their morphological similarity; they were assigned to family Podoviridae, order Caudovirales on the basis of their head and tail morphology. Structural proteins resolved on SDS-PAGE showed the presence of similar major outer membrane proteins. The bacteriophages, belonging to Podoviridae family with short stumpy tails, were found to be nontoxic to mice. They showed maximum count in various organs at 6 h post inoculation, which persisted till 36 h. These phages thus have the potential to be used for phage therapy.

• MeSH
• anatomické struktury zvířat virologie   MeSH
• bakteriofágy MeSH
• DNA fingerprinting MeSH
• DNA virů genetika   MeSH
• DNA genetika   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• financování organizované MeSH
• genetická variace MeSH
• genotyp MeSH
• Klebsiella pneumoniae virologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• odpadní vody virologie   MeSH
• Podoviridae MeSH
• přichycení viru MeSH
• technika náhodné amplifikace polymorfní DNA MeSH
• transmisní elektronová mikroskopie MeSH
• virion ultrastruktura   MeSH
• virové proteiny analýza   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH