江西农业大学学报2024，Vol.46Issue(3)：713-725,13.DOI:10.3724/aauj.2024064
大肠杆菌烈性噬菌体vB_EcoM_JN03的分离、鉴定及其抑菌效果研究
Isolation,identification and antibacterial effect study of E.coli lytic bacteriophage vB_EcoM_JN03
摘要
Abstract
[Objective]E.coli O157:H7 is one of important foodborne pathogens.The improper use of antibiotics has led to E.coli O157:H7 multi-drug resistance and pan-drug resistance becoming an increasingly serious problem.The study of its natural bactericide,virulent phage,has theoretical significance and practical application value for the prevention and control of E.coli O157:H7 pollution.[Method]In this study,the multi-drug resistant E.coli O157:H7 was used as the host bacteria,and the virulent phage was isolated from sewage by double-layer agar plate method.Its biological characteristics were characterized and the whole genome sequence was analyzed.The antibacterial effect of the phage on E.coli O157:H7 in food was evaluated.[Result]A phage capable of lysing E.coli O157:H7 was successfully isolated and purified,named vB_EcoM_JN03(GenBank accession number:OR885871).vB_EcoM_JN03 belongs to Caudovirales,Myoviridae.The optimal multiplicity of infection(MOI)was 0.01,the incubation period was 30 min,and it reached the plateau after 120 min.The burst size was 80 PFU/cell.It is stable in the range of 30-60℃ and pH 4-12.The genome of bacteriophage vB_EcoM_JN03 is double-stranded DNA,with a total length of 158142 bp,G+C content of 44.63%,encoding 206 open reading frame(ORF),and does not contain known encoded virulence,antibiotic resistance and lysogenic genes.vB_EcoM_JN03 significantly inhibited E.coli O157:H7 in milk and beef samples at 4℃.[Conclusion]In this study,a virulent bacteriophage vB_EcoM_JN03 of E.coli O157:H7 was isolated and identified.It has good inhibitory effect on host bacteria and is expected to be used as a natural bactericide to control E.coli O157:H7 contaminated food.关键词
大肠杆菌O157:H7/烈性噬菌体/生物学特性/生物杀菌剂Key words
E.coli O157:H7/lytic phage/biological characterization/biological fungicide分类
轻工纺织引用本文复制引用
李天娇,舒梅,吴国平,钟婵,潘红,赵远洋,张玲..大肠杆菌烈性噬菌体vB_EcoM_JN03的分离、鉴定及其抑菌效果研究[J].江西农业大学学报,2024,46(3):713-725,13.基金项目
国家自然科学基金地区项目(32160608、32360629)和江西省现代农业产业技术体系建设专项(JXARS-12)Project supported by the Regional Project of National Natural Science Foundation of China(32160608、32360629)and Jiangxi Province Modern Agricultural Industrial Technology System Construction Special Fund(JXARS-12)江西省科学技术支撑项目(20212BBF63027,2023BAB205080)同时对本研究给予了资助,谨致谢意! （32160608、32360629）
