Isolation and identification of a novel bovin Klebsiella pneumoniae bacteriophage vB_Kpn_B01

Abstract:

[Background] Klebsiella pneumoniae (K. pneumoniae) is an important pathogen that can infect both humans and bovine. The wide spread of Klebsiella pneumoniae and its high drug resistance have caused great difficulties in the treatment of the disease. [Objective] To isolate a lytic Klebsiella pneumoniae bacteriophage. Its biological properties were studied, and the entire genome analysis of the bacteriophage was performed. [Methods] Klebsiella pneumoniae bacteriophage was isolated by the double-layer agar culture method from a dairy farm trough in Sichuan province. We used the same method to conduct host range, thermostability, pH, optimal multiplicity of infection (MOI), and one-step growth experiment. Phage morphology was observed by transmission electron microscopy (TEM). The whole genome of the bacteriophage was sequenced. [Results] A lytic Klebsiella pneumoniae bacteriophage vB_Kpn_B01 was isolated which had strong specificity. The morphology of plaque was round, transparent plaques with clear boundaries. Phage found in this study was double-stranded DNA bacterial viruses belonging to the Siphoviridae family, and the size of it was 113 227 bp. Comparative genome analysis revealed that vB_Kpn_B01 genome possesses the highest similarity to the bacteriophages in the genus of Sugarlandvirus. The genome of vB_Kpn_B01 comprises 149 coding sequences (CDS) and 25 tRNAs, but does not contain any known antibiotic resistant genes or virulent genes. [Conclusion] Phage vB_Kpn_B01 has a strong lysis ability and the necessary genes to lyse host bacteria. It has the potential to be used in the prevention and treatment of multi-drug resistant bacteria in animal husbandry.