中国兽医学报2024,Vol.44Issue(8) :1691-1697.DOI:10.16303/j.cnki.1005-4545.2024.08.15

一株产气荚膜梭菌烈性噬菌体的分离鉴定及特性分析

Isolation,identification and characterization of a virulent phage of Clostridium perfringens

李梦娇 宋战昀 刘博 许智强 刘月 王秋霖 冯新
中国兽医学报2024,Vol.44Issue(8) :1691-1697.DOI:10.16303/j.cnki.1005-4545.2024.08.15

一株产气荚膜梭菌烈性噬菌体的分离鉴定及特性分析

Isolation,identification and characterization of a virulent phage of Clostridium perfringens

李梦娇 1宋战昀 2刘博 2许智强 1刘月 1王秋霖 1冯新1
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作者信息

  • 1. 吉林大学动物医学学院人兽共患病研究教育部重点实验室,吉林长春 130062
  • 2. 长春海关技术中心,吉林长春 130062
  • 折叠

摘要

产气荚膜梭菌(Clostridium perfringens)是一种常见的革兰阳性厌氧条件致病菌,在自然界中广泛存在,可引起腹泻、气性坏疽等疾病.临床上采用抗生素治疗产气荚膜梭菌感染,但细菌会通过突变、耐药质粒传播等方式产生耐药性,使产气荚膜梭菌在抗生素的环境压力下得以存活.因此寻找和开发新制剂来替代抗生素或作为饲料添加剂以定向清除机体携带的产气荚膜梭菌或预防感染是十分重要的.本研究从污水中分离得到了一株产气荚膜梭菌烈性噬菌体vB_CPP_AT.利用透射电镜观察噬菌体形态,通过裂解谱、MOI、酸碱及温度耐受性等分析该产气荚膜梭菌噬菌体基本生物学特性.研究发现vB_CPP_AT噬菌体为短尾噬菌体,在60 min时呈暴发式增长,最佳MOI为0.1,专一性裂解产气荚膜梭菌,裂解率为40%(8/20),与参试的其他细菌不发生裂解反应,具有良好的热稳定及酸碱耐受性.vB_CPP_AT噬菌体基因组为双链DNA,全长16 790 bp,具有20个开放阅读框.基因组分析vB_CPP_AT噬菌体为1株新的产气荚膜梭菌烈性噬菌体.该结果为噬菌体vB_CPP_AT应用于产气荚膜梭菌的临床治疗奠定了基础.

Abstract

Clostridium perfringens is a common Gram-positive anaerobic conditioned pathogen,widely existing in nature,which can cause diarrhea,gas gangrene,and other diseases.Antibiotics are used in the clinical treatment of Clostridium perfringens infection,but the bacteria will devel-op resistance through mutation,drug-resistant plasmid transmission,and other ways,so that Clos-tridium perfringens can survive under the environmental pressure of antibiotics.Therefore,it is very important to find and develop new preparations to replace antibiotics or as feed additives to target the removal of Clostridium perfringens from the body or to prevent infection.In this study,a virulent Clostridium perfringens phage vB_CPP_AT was isolated from sewage by double plate method.The morphology of the bacteriophage was observed by transmission electron microscope.The biological characteristics of the bacteriophage were analyzed by lytic spectrum,MOI,pH,and temperature tolerance.The results showed that the vB_CPP_AT belongs to the Podoviridae.It would grow explosively at 60 min with an optimal MOI of 0.1.The vB_CPP_AT only lyse Clos-tridium perfringens and the lytic rate was 40%(8/20).No cleavage reaction occurred with other bacteria tested.The phage had good thermal stability and acid-base tolerance.Genomic analysis re-vealed that the phage had double-stranded DNA with a total length of 16 790 bp,and 20 open read-ing frames.Genomic analysis of vBCPPAT showed that it was a new virulent phage of Clostridi-um perfringens.The results laid a foundation for the clinical treatment of Clostridium perfringens with phage.

关键词

产气荚膜梭菌/噬菌体生物学特性/基因组

Key words

Clostridium perfringens/biological characteristics of phage/genome

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基金项目

吉林省教育厅基金资助项目(JJKH20231204KJ)

国家自然科学基金资助项目(U19A2038)

出版年

2024
中国兽医学报
吉林大学

中国兽医学报

CSTPCDCSCD北大核心
影响因子:0.702
ISSN:1005-4545
参考文献量7
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