沙门菌烈性噬菌体的分离鉴定、生物学特性及基因组分析

doi:10.11843/j.issn.0366-6964.2022.08.025

摘要/Abstract

摘要： 旨在为研制沙门菌特异性抗菌制剂提供试验数据和参考，以禽源沙门菌为宿主菌，从鸡场污水中分离纯化沙门菌噬菌体，并对其生物学特性和基因组学进行研究。结果显示：分离到1株沙门菌烈性噬菌体S5，透射电镜下该噬菌体头部直径约50 nm，尾部长约200 nm，头部呈正二十面体对称，属有尾噬菌体目、长尾噬菌体科；该噬菌体对20株沙门菌的裂解率为75%，效价为8×10⁹PFU·mL^-1；在30~60℃作用30 min和1 h的效价基本不变；pH 4.0~11.0对噬菌体活性影响不明显；最佳感染复数为0.000 1，一步生长曲线显示其感染宿主菌后5 min内（潜伏期）效价基本保持不变，暴发期约为95 min，平均裂解量为50 PFU·cell^-1。噬菌体S5是双股DNA病毒，基因组全长为72 519 bp，GC含量39.45%，含有9种tRNA，预测到109个ORF，未发现任何编码细菌毒力因子等有害产物的基因。沙门菌烈性噬菌体S5，裂解效果较好、对环境适应性强、遗传背景清楚，以期成为防控沙门菌的抗生素替代产品。

关键词: 沙门菌, 噬菌体, 生物学特性, 生物信息学分析, 基因组

Abstract: The current study aims to provide experimental data and references for the development of Salmonella specific antibacterial agents. Here, we isolated a Salmonella phage from chicken farm sewage and studied its biological characteristics and genomics. The results showed that Salmonella phage S5 was isolated. Transmission electron microscopy showed that the phage was a long-tailed phage with a head diameter of about 50 nm and a tail length of about 200 nm. Its head was icosahedral symmetrical, belonging to Caudovirales, Siphoviridae. The phage has a lysis rate of 75% against 20 strains of Salmonella, with a titer of 8×10⁹ PFU·mL^-1. The phage titer remained basically unchanged at the temperature from 30 to 60℃ for 30 min or 1 h, with little effect on phage activity at pH4.0-11.0. The optimal MOI of phage was 0.000 1. The one-step growth curve showed that its titer remained basically unchanged within 5 min incubation period after infection with the host bacteria. The outbreak period was about 95 min, and the average lysate was 50 PFU·cell^-1. Phage S5 was a double-stranded DNA virus. The genome was 72 519 bp in length, and the GC content was 39.45%. It contained 9 types of tRNAs and 109 predicted ORFs. No harmful factors such as bacterial virulence factors had been found. Salmonella virulent phage S5 has good lysis effect, strong adaptability to the environment and clear genetic background. It is expected to become an alternative product of antibiotics for the prevention and control of Salmonella.

Key words: Salmonella, phage, biological characteristics, bioinformatics analysis, genome

中图分类号:

Q939.48

岑鑫, 阳亭亭, 赵尊福, 文永平, 张焕容. 沙门菌烈性噬菌体的分离鉴定、生物学特性及基因组分析[J]. 畜牧兽医学报, 2022, 53(8): 2677-2688.

CEN Xin, YANG Tingting, ZHAO Zunfu, WEN Yongping, ZHANG Huanrong. Isolation, Identification, Biological Characteristics and Genome Analysis of a Virulent Salmonella Phage[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2677-2688.

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