阪崎克罗诺杆菌噬菌体JC01的筛选和特性  

Isolation and characterization of bacteriophage JC01 against Cronobacter sakazakii

作　　者：刘丹丹[1] 兰冠达 蒋洁 李菁华[1] 黄红兰[1] 赵春燕[1] LIU Dandan;LAN Guanda;JIANG Jie;LI Jinghua;HUANG Honglan;ZHAO Chunyan(Department of Pathogen Biology,College of Basic Medical Science,Jilin University,Changchun 130021,Jilin,China;Department of Laboratory Medicine,The People’s Hospital of Guangxi Zhuang Autonomous Region(Guangxi Academy of Medical Sciences),Nanning 530021,Guangxi,China)

机构地区：[1]吉林大学基础医学院病原生物学系,吉林长春130021 [2]广西壮族自治区人民医院(广西医学科学院)检验科,广西南宁530021

出　　处：《微生物学通报》2023年第12期5475-5486,共12页Microbiology China

基　　金：吉林省科技厅国际合作项目(20210402023GH)。

摘　　要：【背景】阪崎克罗诺杆菌是一种食源性病原体,摄入受污染的婴儿配方奶粉(powdered infant formula,PIF)常引起新生儿坏死性小肠结肠炎和脑膜炎,对早产儿和免疫功能低下的婴儿健康甚至生命产生严重威胁。噬菌体具有特异性杀菌性,可成为一种新型生物防控制剂预防阪崎克罗诺杆菌和其他食源性致病菌的污染。【目的】从污水中分离出感染阪崎克罗诺杆菌噬菌体,评价其生物学特性、基因组生物信息及在婴儿配方奶粉中杀菌作用。【方法】采用双层琼脂法分离鉴定噬菌体,并测定其酸碱稳定性、温度稳定性、宿主范围和一步生长曲线,透射电镜观察形态,对其基因组进行二代测序和裂解功效测定。【结果】从污水中分离到一株新的能裂解阪崎克罗诺杆菌的噬菌体JC01,具有二十面立体对称头部和非收缩的长尾,噬菌体JC01基因组为双链DNA,由61736 bp组成,预测有76个开放阅读框(open reading frame,ORF),不含有tRNA,基因组中不含有耐药基因和毒力基因。系统发育分析及基因比对分析显示噬菌体JC01是全新的噬菌体,归类于有尾噬菌体纲(Caudoviricetes)卡金斯病毒科(Casjensviridae)雅昆病毒属(Jacunavirus),被命名为Jacunavirus 01。噬菌体JC01在不同温度(−20–40°C)和pH 5.0–9.0范围内稳定,且对婴儿配方奶粉污染的阪崎克罗诺杆菌具有良好的杀菌效果。【结论】JC01噬菌体是裂解阪崎克罗诺杆菌的新噬菌体,作为生物安全防控剂在食品生产和加工方面具有很大潜力。[Background]Cronobacter sakazakii is an emerging food-borne pathogen.The powdered infant formula(PIF)contaminated by this pathogen may cause necrotizing enterocolitis and meningitis in infants,threatening the life of premature or immunocompromised infants.Bacteriophages are well recognized for their antibacterial properties and may be novel biocontrol agents for C.sakazakii and other food-borne pathogenic bacteria.[Objective]To characterize a novel C.sakazakii bacteriophage isolated from sewage and evaluate its antibacterial effects in PIF.[Methods]The double-layer agar method was used to isolate phages,and then the pH stability,thermal stability,host range,one-step growth curve of the isolate were determined.The phage morphology was observed by transmission electron microscopy.Next-generation sequencing was performed for the genome of the phage,and the lysis efficiency was evaluated.[Results]A novel bacteriophage JC01 against C.sakazakii was isolated from sewage.The phage had an icosahedral head and a long non-contractile tail.The genome of JC01 was 61736 bp in length,containing 76 the open reading frames(ORF)and no drug-resistance gene,virulence gene or tRNA.The phylogenetic analysis and genome comparison indicated that JC01 was a novel phage belonging to the genus Jacunavirus,the class Casjensviridae of the order Caudoviricetes,and its was named as Jacunavirus JC01.The phage was stable at−20 to 40°C and pH 5.0−9.0 and capable of lysing C.sakazakii in the contaminated PIF.[Conclusion]JC01 appears to be a novel phage against C.sakazakii,and it might be an alternative biocontrol agent for C.sakazakii contamination in food processing and production environments.

关 键 词：阪崎克罗诺杆菌 噬菌体 基因组分析 食品安全 

分 类 号：S852.61[农业科学—基础兽医学]