实验环境中野生型鼠疫噬菌体的生长表型特征  

Phenotypic characteristics of wild-type plague phage growth in different experimental environments

作　　者：李存香[1] 祁芝珍[1] 张青雯[1] 冯建萍[1] 金泳[1] 赵海红[1] 应凯业 张晓璐 李广辉 金星[1] LI Cunxiang;QI Zhizhen;ZHANG Qingwen;FENG Jianping;JIN Yong;ZHAO Haihong;YING Kaiye;ZHANG Xiaolu;LI Guanghui;JIN Xing(Qinghai Institute for Endemic Disease Control and Prevention,Xi'ning,Qinghai 810021,China)

机构地区：[1]青海省地方病预防控制所,鼠疫菌专业实验室,青海西宁811602

出　　处：《中国热带医学》2023年第7期720-724,共5页China Tropical Medicine

基　　金：青海省医药卫生科技项目重点课题(No.2022-wjzd-06)。

摘　　要：目的 观察野生型鼠疫噬菌体在不同实验培养环境中的生长表型特征,为后续噬菌体的生物学特性鉴定及其与宿主菌相互作用研究提供科学依据。方法 利用液体培养法、固体培养法和基于OmniLogTM微生物鉴定系统的微量液体培养法检测3株野生型鼠疫噬菌体的生长情况。结果 基于LB液体培养基的生长结果显示,鼠疫噬菌体476号经28℃和37℃作用20~24 h后生长状况优于37℃培养的鼠疫噬菌体087号和072204号,37℃培养的鼠疫噬菌体087号优于072204号。以鼠疫疫苗株EV76为宿主菌,鼠疫噬菌体476号在双层琼脂培养基上经28℃和37℃培养20~24 h后产生较为清晰的噬菌斑;鼠疫噬菌体087号和072204号经37℃培养20~24 h后在双层琼脂培养基上产生不透明的噬菌斑。基于OmniLog TM系统微量液体培养法的生长结果显示,3株鼠疫噬菌体经33℃裂解鼠疫疫苗株EV76时,通过96孔微量板曲线图显示第1列出现横线,且峰值不超过100,随着噬菌体数量的降低,稀释试验孔依次均出现与鼠疫疫苗株EV76相似的生长曲线,实验孔中四唑类染料颜色随着噬菌体数量的减少和宿主菌数量的增多而逐渐加深。由此说明鼠疫噬菌体476号在28℃和37℃均裂解鼠疫疫苗株EV76;鼠疫噬菌体087号和072204号在37℃均裂解鼠疫疫苗株EV76。结论 3株野生型鼠疫噬菌体的裂解能力具有温度依赖性,3种实验条件下的生长结果一致。Objective To observe the phenotypic characteristics of 3 wild-type plague phages under different experimental environments,providing scientific evidence for the identification of phage biological characteristics and the study of their interaction with host bacteria in the future.Methods The sensitivity of 3 wild-type plague phages were detected by using liquid culture method,emisolid medium method and micro-liquid culture method based on OmniLog TM microbial identification system.Results The growth result based on LB liquid medium showed that the growth of plague phage 476 for 20-24 hours at both 28℃and 37℃ was better than that of plague phages 087 and 072204 at 37℃,and the growth of plague phages 087 was better than that of plague phages 072204 at 37℃.With the attenuated plague bacterium EV76 as the host bacterium,phage 476 was able to form visible plaque on double-layer agar medium for 20-20 hours at both 28℃ and 37℃,phages 087 and 072204 were only able to form opaque plaque on double-layer agar medium for 20-24 hours at 37℃.The growth results based on OmniLogTM system showed that when plague phage was lysed in EV76 strain at 33℃,the first row appeared as a straight line with a peak of no more than 100 in the 96-well microplate curve chart.As the phage quantity decreased,the dilution plate appeared with growth curve similar to EV76 strain in turn,and the color of tetrazolium dyes in the experimental wells gradually deepened as the phage number decreased and the host bacteria number increased.Therefore,it indicates that phage 476 was sensitively at both 28℃and 37℃,while phage 087 and 072204 were temperature-dependent only at 37℃to attenuated plague bacterium EV76.Conclusions The lysing ability of 3 wild-type plague phages are temperature-dependent,and the growth results are consistent under the three experimental conditions.

关 键 词：鼠疫噬菌体 野生型 实验环境 表型特征 

分 类 号：R378[医药卫生—病原生物学]