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鼠伤寒沙门菌烈性噬菌体的分离鉴定与生物学特性
引用本文:何依蓉,张奕杰,杨伟,陈晓聪,张家翔,陈培富. 鼠伤寒沙门菌烈性噬菌体的分离鉴定与生物学特性[J]. 畜牧兽医学报, 2021, 52(3): 763-771. DOI: 10.11843/j.issn.0366-6964.2021.03.020
作者姓名:何依蓉  张奕杰  杨伟  陈晓聪  张家翔  陈培富
作者单位:云南农业大学动物医学院, 昆明 650201
基金项目:云南省专家工作站项目(202005AF150041);国家级大学生创新创业项目(201810676018)。
摘    要:
旨在分离鼠伤寒沙门菌烈性噬菌体,为控制该病原菌感染或污染提供生物制剂.以1株从市售散装牛奶中分离到的多重耐药性鼠伤寒沙门菌为宿主菌,采用人工诱导方法,连续1周每日给3只试验鸡饲喂宿主菌悬液,7 d后,采用双层琼脂平板法从试验鸡粪便中分离培养噬菌体,并对其效价、核酸类型、宿主谱、热稳定性与酸碱耐受性以及对鼠伤寒沙门菌感染...

关 键 词:鼠伤寒沙门菌  烈性噬菌体  生物学特性  生物防治
收稿时间:2020-08-25

Isolation,Identification and Biological Properties of a Lytic Phage against Salmonella Typhimurium
HE Yirong,ZHANG Yijie,YANG Wei,CHEN Xiaocong,ZHANG Jiaxiang,CHEN Peifu. Isolation,Identification and Biological Properties of a Lytic Phage against Salmonella Typhimurium[J]. Chinese Journal of Animal and Veterinary Sciences, 2021, 52(3): 763-771. DOI: 10.11843/j.issn.0366-6964.2021.03.020
Authors:HE Yirong  ZHANG Yijie  YANG Wei  CHEN Xiaocong  ZHANG Jiaxiang  CHEN Peifu
Affiliation:College of Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, China
Abstract:
Isolation of an appropriate lytic phage against Salmonella Typhimurium will help to provide a biological for controlling infection or pollution caused by this pathogen.A strain of Salmonella Typhimurium with multidrug resistance which had been isolated from bulk milk was used as host bacteria.Artificial induction method was employed.Three chickens were fed daily with the host bacterium suspension for a week.After 7 days,the phage isolation and culture was conducted from chicken feces by using double agar plate method.The titer,nucleic acid type,host spectrum,thermal and acid-base stability and therapeutic effect in Salmonella Typhimurium infected mice using the phage were analyzed.A lytic phage against Salmonella Typhimurium was isolated and purified,named KM104.This phage efficiently lysed the homologous host strain and another Salmonella Typhimurium strains in vitro,producing clear and circular plaques with a diameter of about 1 mm.The viral nucleic acid type was DNA and its genome size was 28 kb or so.The highest titer of the virus was 4.6×108 PFU·mL-1.The optimal multiplicity of infection(MOI)was 1:10,the incubation period was approximately 6 min,and it reached the highest titer at 70 min.The phage maintained high viability under the condition of 20-60℃and a pH value range of 2.0-8.0.The phage effectively reduced the injury in duodenal tissue of experimental mice caused by Salmonella Typhimurium.The results here indicated that KM104 can efficiently lyse Salmonella Typhimurium in vivo and in vitro with rapid proliferation and strong adaptability to the environment.Its acid-fast character is expected to help resist the degradation by gastric acid,making the phage have a potential application value in preventing and controlling Salmonella Typhimurium infection or pollution.
Keywords:Salmonella Typhimurium  lytic phage  biological properties  biological control
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