1株沙门氏菌噬菌体的分离鉴定和特性分析

doi:10.16431/j.cnki.1671-7236.2023.10.031

中国畜牧兽医 ›› 2023, Vol. 50 ›› Issue (10): 4168-4178.doi: 10.16431/j.cnki.1671-7236.2023.10.031

1株沙门氏菌噬菌体的分离鉴定和特性分析

廖彬如¹, 余春燕¹, 黄巧辰¹, 莫玉鹏^1,2, 黄炜乾^1,3, 李磊¹, 韩凯欧¹, 王乐平¹, 李珣¹, 王晓晔¹

1. 广西大学动物科学技术学院, 广西壮族自治区兽用生物制品工程研究中心, 广西畜禽繁育与疾病防控重点实验室, 广西高校动物疫病预防与控制重点实验室, 南宁 530004;
2. 桂林力源集团, 桂林 541012;
3. 广东容大生物股份有限公司, 清远 515000

收稿日期:2023-04-25 出版日期:2023-10-05 发布日期:2023-09-26
通讯作者: 王晓晔 E-mail:xywang@gxu.edu.cn
作者简介:廖彬如,E-mail:1906369437@qq.com;余春燕,E-mail:1415733127@qq.com
基金资助:
南宁市重点研发计划(20212023)；良庆区重点研发计划(202109)；防城港市重点研发计划(AB21014016)；武鸣区重点研发计划(20210102)；江南重点研发计划(20220620-2)

Isolation,Identification and Characterization of a Strain of Salmonella Phage

LIAO Binru¹, YU Chunyan¹, HUANG Qiaochen¹, MO Yupeng^1,2, HUANG Weiqian^1,3, LI Lei¹, HAN Kaiou¹, WANG Leping¹, LI Xun¹, WANG Xiaoye¹

1. Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics, Guangxi Key Laboratory of Animal Reproduction, Breeding and Disease Control, Guangxi Colleges and Universities Key Laboratory of Prevention and Control for Animal Disease, College of Animal Science and Technology, Guangxi University, Nanning 530004, China;
2. Guilin Liyuan Group, Guilin 541012, China;
3. Guangdong Rongda Biological Co., Ltd., Qingyuan 515000, China

Received:2023-04-25 Online:2023-10-05 Published:2023-09-26

摘要/Abstract

摘要： 【目的】分离有效裂解沙门氏菌的噬菌体,为探索新的沙门氏菌防控方法提供理论依据和参考。【方法】以鸡白痢沙门氏菌X-1014为宿主菌,通过双层琼脂平板法从污水中分离并纯化出噬菌体。采用噬菌斑与电子显微镜观察、宿主谱测定、生物学特性测试、体外抑菌试验及噬菌体的全基因组测序分析方法评估该噬菌体。【结果】分离纯化出1株能裂解沙门氏菌和大肠杆菌的裂解性噬菌体,命名为ZH5(GenBank登录号:OM864357.1)。双层平板法结果显示,噬菌斑透亮且边缘清晰。电镜观察显示,噬菌体ZH5头部呈二十面体、头部直经约50 nm,尾部长度约20 nm。宿主谱分析结果显示,噬菌体ZH5能裂解3株沙门氏菌和1株大肠杆菌。生物学特性结果显示,最佳感染复数为10,潜伏期和爆发期分别为20和60 min,裂解量约为139 PFU/cell;酸碱耐受范围为pH 4.0~10.0,且在温度为4~50 ℃的环境中能存活30 min。体外抑菌结果显示,当感染复数为10时,噬菌体ZH5对沙门氏菌X-1014和CVCC 1806抑制效果最佳。全基因组分析结果显示,该噬菌体全长为42 949 bp,GC含量为51.41%,其中已知功能编码序列(coding sequences,CDS)占比为52%。线性比对分析结果显示,噬菌体ZH5与大肠杆菌噬菌体 Minorna相似性最高(相似性94.19%;覆盖率84%)。系统发育进化树分析表明,噬菌体ZH5属于Drulisvirus属。【结论】本研究分离出1株能跨宿主裂解沙门氏菌和大肠杆菌的噬菌体,为进一步研究沙门氏菌抗菌剂提供了良好的材料。

关键词: 沙门氏菌; 噬菌体; 生物学特性; 体外抑菌效果; 全基因组分析

Abstract: 【Objective】 It was aimed to isolate a phage with effective cracking for Salmonella,and provide experimental data and reference for exploring the countermeasure method of Salmonella.【Method】 Salmonella Pullorum X-1014 was used as the host bacteria,and the phage was isolated and purified from sewage by double-layer plate method.The phage was evaluated by plaque and electron microscopy,host spectrum,biological characteristics test,antibacterial test in vitro and whole genome sequencing analysis.【Result】 A lytic phage was isolated and purified,named ZH5 (GenBank accession No.:OM864357.1),which could lyse Escherichia coli and Salmonella.The results of double-layer plate method showed that the plaque was transparent and the edge was clear.Electron microscopy showed that phage ZH5 had an icosahedral head with a straight diameter of about 50 nm,a tail with a length of about 20 nm.The phage ZH5 could lyse three strains of Salmonella and one strain of Escherichia coli.The results of biological characteristics showed that the optimal multiplicity of infection was 10,the incubation period was 20 min and the outbreak period was 60 min,and the lysis volume was about 139 PFU/cell.The acid-base tolerance range was pH 4.0-10.0,and it could survive for 30 min in the environment of 4-50 ℃.Phage ZH5 showed the best inhibitory effect on Salmonella X-1014 and CVCC 1806 when the multiplicity of infection was 10.The whole genome analysis showed that the length of the phage was 42 949 bp,and the GC content was 51.41%,of which the proportion of known functional coding sequences (CDS) was 52%.Phage linear alignment showed that the genome of phage ZH5 was the most similar to the Escherichia phage Minorna(identity 94.19%;cover 84%).Phylogenetic tree analysis showed that phage ZH5 belonged to the Drulisvirus genus.【Conclusion】 A phage capable of cross-host lysis of Salmonella and Escherichia coli was isolated,which provided a good theoretical material for further research on Salmonella antibacterial agents.

Key words: Salmonella; phage; biological characteristics; in vitroantibacterial effect; whole-genome analysis

中图分类号:

S852.61

廖彬如, 余春燕, 黄巧辰, 莫玉鹏, 黄炜乾, 李磊, 韩凯欧, 王乐平, 李珣, 王晓晔. 1株沙门氏菌噬菌体的分离鉴定和特性分析[J]. 中国畜牧兽医, 2023, 50(10): 4168-4178.

LIAO Binru, YU Chunyan, HUANG Qiaochen, MO Yupeng, HUANG Weiqian, LI Lei, HAN Kaiou, WANG Leping, LI Xun, WANG Xiaoye. Isolation,Identification and Characterization of a Strain of Salmonella Phage[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4168-4178.

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1株沙门氏菌噬菌体的分离鉴定和特性分析

Isolation,Identification and Characterization of a Strain of Salmonella Phage

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