一株鼠伤寒沙门氏菌噬菌体的分离鉴定及对小鼠的防制效果研究

doi:10.16431/j.cnki.1671-7236.2018.01.030

摘要/Abstract

摘要：

本研究旨在通过双层平板法从鸡粪中分离鼠伤寒沙门氏菌烈性噬菌体，并通过电镜观察、热稳定性、pH稳定性、最佳感染复数、一步生长曲线和在小鼠上的防制效果试验对噬菌体进行评估。结果显示，从鸡粪中成功分离出一株鼠伤寒沙门氏菌噬菌体（φBLCC8-0050-3），该噬菌体有一个正多面体立体对称的头部，头径直径约65 nm，无尾部结构，应属于盖噬菌体科（Tectiviridae）烈性噬菌体；噬菌斑成圆形空斑，周围有晕环，直径为2~3 mm；温度耐受范围是30~50℃；pH耐受范围为3~10。当感染复数（multiplicity of infection，MOI）为0.0001时，子代噬菌体滴度最高；按照一步生长曲线结果，潜伏期为20 min左右，爆发时间为180 min左右，爆发量为43 333 PFU/cell；从噬菌体对鼠伤寒沙门氏菌的防制效果来看，处理14 d后，阳性对照组小鼠存活率为0，噬菌体预防组小鼠存活率为90%，噬菌体治疗组小鼠存活率为70%。说明与阳性对照组相比，预防组和治疗组都对小鼠抵抗沙门氏菌感染有一定的作用。结果表明，φBLCC8-0050-3是一株具有良好应用前景的噬菌体，可作为一种生物抑菌剂用于鼠伤寒沙门氏菌的防制。

关键词: 鼠伤寒沙门氏菌; 裂解性噬菌体; 生物学特性; 防制效果

Abstract:

This study was aimed to separate a lytic phage against Salmonella Typhimurium from the manure of chicken using double-layer agar culture method. The lytic phage was assessed according to electron microscope observation, thermal stability, pH stability, optimal multiplicity of infection (MOI), One-step growth curve and the inactivation effects on the growth of Salmonella Typhimurium were assayed. As a result, a lytic phage (φBLCC8-0050-3) against Salmonella Typhimurium was successfully isolated from the manure of chicken.φBLCC8-0050-3 had an icosahedral head approximately 65 nm in diameter and had no tail. φBLCC8-0050-3 belonged to family Tectiviridae according to the international classification standard. The plaques of φBLCC8-0050-3 were round and had halo ring around and their sizes which was about 2 to 3 mm in diameter. φBLCC8-0050-3 was stable over a range of temperature (30 to 50℃) and pH (3 to 10). The optimal MOI of φBLCC8-0050-3 was 0.0001. One-step growth experiment showed that the latent time and burst time were 20 and 180 min, respectively, and the burst size was 43 333 PFU/cell. From the control effect of bacteriophage against Salmonella Typhimurium, after 14 days of treatment, the survival rate of mice in the positive control group was 0, was 90% in phage prevention group, and was 70% in treatment group. Compared with the positive control group, the prevention group and the treatment group had a certain effect on mice against Salmonella infection. The results showed that the φ BLCC8-0050-3 was a promising phage, which could be used as a biological antibacterial agent in the prevention and control of Salmonella Typhimurium.

Key words: Salmonella Typhimurium; lytic phages; biological properties; control effect

中图分类号:

S852.61

赵影, 刘秀侠, 李金敏, 徐海燕, 谷魏. 一株鼠伤寒沙门氏菌噬菌体的分离鉴定及对小鼠的防制效果研究[J]. 《中国畜牧兽医》, 2018, 45(1): 234-241.

ZHAO Ying, LIU Xiuxia, LI Jinmin, XU Haiyan, GU Wei. Isolation and Identification of a Phage Against Salmonella Typhimurium and Its Control Effect on Mice[J]. , 2018, 45(1): 234-241.

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