中国畜牧兽医2023，Vol.50Issue(12)：5160-5169,10.DOI:10.16431/j.cnki.1671-7236.2023.12.037
1株肠炎沙门菌噬菌体GD01的分离鉴定及生物学特性分析
Isolation,Identification and Biological Characterization Analysis of a Bacteriophage GD01 from Salmonella Enteritidis
摘要
Abstract
[Objective]The objective of this study was to analyze the biological characteristics and genomic information of a phage of Salmonella Enteritidis isolated and purified from sewage.[Method]Using Salmonella Enteritidis as host,a phage of Salmonella Enteritidis was isolated and purified from mixed samples of lake water from South China Agricultural University and sewage from a broiler farm in Guangzhou.The host spectrum,optimal multiplicity of infection,one-step growth curve,thermal stability and pH stability of phage were detected.The genome of the bacteriophage was sequenced and bioinformatics analyzed,and the bacteriostatic ability of the bacteriophage against Salmonella in milk contaminated at different temperatures was investigated.[Result]A phage of Salmonella Enteritidis was successfully isolated and named GD01,which could form round,transparent and clearly edged plaque on a double-layer agar plate.Under electron microscopy,the head of GD01 was icosahedral symmetry,with a diameter of 62 nm and a tail length of 224.4 nm,belonged to the Caudovirales,Siphoviridae.The host spectrum was narrow,and the optimum pH was 8.0,the optimum temperature was 37℃,the optimal multiplicity of infection was 0.01.The one-step growth curve showed that the latency of GD01 was 10 min,the lysis period was 110 min,and the average amount of cleavage was 50 PFU/cell.Whole genome sequencing analysis results showed that the total length of GD01 genome was 60 109 bp,the GC content was 56.22％,it was double-stranded DNA,contained 50 open reading frame(ORF),and did not contain virulence genes,drug resistance genes and tRNA genes.The results of in vitro bacteriostatic test showed that phage GD01 still had good bacteriostatic effect on Salmonella after treatment at 37 ℃ for 8 h and at 4 ℃ for 72 h.[Conclusion]The isolated bacteriophage GD01 of Salmonella Enteritidis had strong acid-base tolerance and temperature tolerance,clear bioinformatics properties,and good antibacterial effect in vitro,which had the potential and application prospect of replacing antibiotics.关键词
沙门菌/噬菌体/生物学特性/全基因组测序Key words
Salmonella/phage/biological characteristics/whole genome sequencing分类
农业科技引用本文复制引用
高铎,王晋宇,李河谕,蒋祺丰,陈子涵,徐禧莹,贾坤..1株肠炎沙门菌噬菌体GD01的分离鉴定及生物学特性分析[J].中国畜牧兽医,2023,50(12):5160-5169,10.基金项目
广州市科技计划项目(SL2022E04J00201、202206010131、202202010033) （SL2022E04J00201、202206010131、202202010033）
广东省现代农业产业技术体系(2023KJ127) （2023KJ127）
