一株高效裂解高致病性副溶血弧菌噬菌体的分离鉴定与表征  

Isolation and Characterization of a Highly Eficient Lytic Phage against Highly Pathogenic Vibrio parahaemolyticus

作　　者：张永记 陈娅帆 邵伟 刘杰 张维 李新风 钟明月 何进[1] 杨航[3,4] ZHANG Yongji;CHEN Yafan;SHAO Wei;LIU Jie;ZHANG Wei;LI Xinfeng;ZHONG Mingyue;HE Jin;YANG Hang(National Key Laboratory of Agricultural Microbiology&Hubei Hongshan Laboratory,College of Life Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;Hubei Ziwey Biotechnology Co.,Ltd,Wuhan 430200,China;Key Laboratory of Virology and Biosafety,Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430071,China;Hubei Jiangxia Laboratory,Wuhan 430200,China)

机构地区：[1]华中农业大学生命科学技术学院,武汉430070 [2]湖北之微生物科技有限公司,武汉430200 [3]中国科学院武汉病毒研究所,武汉430071 [4]湖北江夏实验室,武汉430200

出　　处：《中国生物工程杂志》2024年第8期48-56,共9页China Biotechnology

基　　金：湖北江夏实验室生物安全科技重大项目(JXBS007)资助项目。

摘　　要：目的:分离获得含半透明仔虾病(translucent post-larvae disease,TPD)毒力因子的副溶血弧菌及裂解该弧菌的噬菌体,并评估噬菌体用于防治水产养殖行业弧菌感染的可能性。方法:利用选择平板法和分子生物学技术分离鉴定弧菌菌株,然后通过双层平板法分离获得噬菌体,并对其进行表征和全基因组测序分析。结果:分离到一株含TPD毒力因子的副溶血弧菌VP392和可裂解该菌株的噬菌体BP471,该噬菌体感染宿主菌最佳感染复数(multiplicity of infection,MOI)为1,潜伏期为20 min,裂解量为181 PFU/cell;在pH 3~11、温度55℃以下、盐浓度50 g/L以下均具有良好的稳定性,可明显抑制宿主菌生长且具有较广的宿主谱;全基因组测序表明其为双链DNA病毒,无毒力因子和抗生素抗性基因。结论:成功分离到一株可裂解含TPD毒力因子的副溶血弧菌噬菌体,其稳定性好、裂解谱广、安全性好,有望作为安全、环保型生物制剂应用到TPD防治中。Objective:To isolate phages with good lytic ability to Vibrio parahaemolyticus with translucent post-larvae disease(TPD)virulence factors and evaluate their potential application in the prevention or treatment of Vibrio infections in aquaculture industry.Methods:Vibrio strains were isolated using selective plates and screened by molecular identification.A phage was separated by double-layer plate method,and evaluated through a series of characterization experiments and whole genome sequencing analysis.Results:Vibrio parahaemolyticus strain VP392 with a translucent virulence factor and a lytic phage BP471 capable of lysing VP392 were isolated.The optimal multiplicity of infection(MOI)of phage BP471 was 1,the latent period was 20 min,and the burst size was 181 PFU/cell;phage BP471 retained good stability at temperatures below 55°C,salt concentration of 50 g/L,and pH 3-11.Phage BP471,characterized as a double-stranded DNA virus devoid of virulence factors and antibiotic resistance genes by whole-genome sequencing analysis,was able to inhibit the growth of its host bacterium in vitro.Conclusion:A lytic phage with good activity against Vibrio parahaemolyticus containing TPD virulence factors was successfully isolated.Phage BP41 has good stability,safety,and broad host spectrum,and may be a safe and environmentally friendly biological agent for the treatment of TPD in aquaculture.

关 键 词：半透明仔虾病 副溶血弧菌 噬菌体 生物学表征 全基因组测序 

分 类 号：Q939[生物学—微生物学]