中国畜牧兽医2024，Vol.51Issue(5)：2047-2057,11.DOI:10.16431/j.cnki.1671-7236.2024.05.026
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
Biological Characteristics and Whole Genome Analysis of a Multidrug-resistant Klebsiella pneumoniae Phage of Sheep Origin
摘要
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.关键词
肺炎克雷伯菌/噬菌体/生物学特性/全基因组分析Key words
Klebsiella pneumoniae/phage/biological characteristics/genomic analysis分类
农业科技引用本文复制引用
侯宫明珠,杨舒涵,邱正青,陈斯奇,周海琴,余星雨,李娜娜,李扬,屈勇刚,李彦芳,梁晏,严多..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[J].中国畜牧兽医,2024,51(5):2047-2057,11.基金项目
兵团重点领域科技攻关计划(2021AB014) （2021AB014）
兵团第十二师重点领域科技攻关计划(SRS2022013) （SRS2022013）
国家自然科学基金(32160847) （32160847）
