1株羊源多重耐药肺炎克雷伯菌噬菌体的生物学特性及全基因组分析

doi:10.16431/j.cnki.1671-7236.2024.05.026

Abstract

Abstract: 【Objective】 The emergence of multidrug-resistant pathogens poses a serious threat to public health security.The biological and genomic characteristics of a lytic phage of multidrug-resistant Klebsiella pneumoniae from sheep were analyzed in order to provide reference for the search of phage preparations for the prevention and treatment of Klebsiella pneumoniae infection.【Method】 Multidrug-resistant Klebsiella pneumoniae strain YH-2 from sheep was used as host bacteria to isolate and purify phage from fecal samples by double agar plate method,then observed the morphology by transmission electron microscope.The host spectrum,optimum multiplicity of infection (MOI),one-step growth curve,thermal stability,acid and base tolerance of phage were measured.The genome characteristics were analyzed based on whole genome sequencing.【Result】 A lytic phage from Klebsiella pneumoniae,named vB_KpnP_PYH-2 (PYH-2),was successfully isolated,which belong to the Podoviridae family.There was a halo around it,the halo spread around as the culture time went on.Phage PYH-2 could not only lyse the Klebsiella pneumoniae strains of sheep origin,but also lyse Klebsiella pneumoniae strains derived from cow mastitis.The optimal MOI was 0.001,the latent period was 15 min,the burst period was 10 min,and the amount of cleavage was 63 PFU/cell.Phage PYH-2 could maintain stable activity at 40-50 ℃ and pH 4.0-12.0.The full length of the phage PYH-2 genome was 45 958 bp,with 51.86% GC content,and it did not contain any drug resistance genes and virulence genes.There were 52 open reading frames (ORFs) in the genome of PYH-2,only 21 ORFs were known functional genes,including 6 tail-associated proteins.【Conclusion】 In this study,a phage with short incubation period,stable biological characteristics and high bactericides was isolated,which could be used as a candidate phage strain of cracking animal derived Klebsiella pneumoniae,laying a foundation for the prevention and treatment of multi-drug resistant Klebsiella pneumoniae infection.

Key words: Klebsiella pneumoniae; phage; biological characteristics; genomic analysis

CLC Number:

S852.61

HOU Gongmingzhu, ZHOU Haiqin, YU Xingyu, LI Nana, LI Yang, QU Yonggang, LI Yanfang, LIANG Yan, YAN Duo, YANG Shuhan, QIU Zhengqing, CHEN Siqi. Biological Characteristics and Whole Genome Analysis of a Multidrug-resistant Klebsiella pneumoniae Phage of Sheep Origin[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2047-2057.

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Biological Characteristics and Whole Genome Analysis of a Multidrug-resistant Klebsiella pneumoniae Phage of Sheep Origin

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