霍氏肠杆菌噬菌体的分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2024.01.032

摘要/Abstract

摘要： 【目的】探究霍氏肠杆菌噬菌体的形态特征和生物学特性，预防和控制霍氏肠杆菌感染。【方法】本研究以霍氏肠杆菌为宿主菌，使用双层琼脂平板法从污水和淤泥中分离噬菌体并进行生物学特性研究。用透射电镜观察纯化后噬菌体的形态特征，测定噬菌体效价、温度稳定性、pH稳定性、最佳感染复数及一步生长曲线。【结果】成功分离鉴定出1株霍氏肠杆菌噬菌体，并命名为H2；该噬菌体在双层琼脂平板上形成大小均一、圆形透明的噬菌斑；透射电镜观察结果显示，噬菌体H2的头部呈二十面体对称，直径为85 nm±2 nm，具有一长尾且有尾丝，尾部长190 nm±2 nm，归属于尾噬菌体目、肌尾噬菌体科；噬菌体H2的温度稳定性和pH稳定性结果显示，在35~55 ℃、pH 5.0~9.0范围内效价基本保持稳定；最佳感染复数为0.0001；根据最佳感染复数绘制一步生长曲线，测得噬菌体H2的潜伏期约为20 min，爆发期约为50 min，平均裂解量约为68 PFU/cell。【结论】噬菌体H2具有裂解能力强、热稳定和酸碱稳定等优点，本研究为后续霍氏肠杆菌噬菌体制剂的研发提供了良好的研究材料。

关键词: 噬菌体; 分离鉴定; 霍氏肠杆菌; 生物学特性

Abstract: 【Objective】 This study was aimed to explore the morphological and biological characteristics of Enterobacter hormaechei phage, and prevent and control Enterobacter hormaechei infections.【Method】 In this study, Enterobacter hormaechei was used as host bacteria.A strain of phage was isolated from sewage and silt by double-layer agar plate method and its biological characteristics were studied.The titer of the phage, temperature and pH stability, optimal multiplicity of infection (MOI) and one-step growth curve of phage were measured.【Result】 A strain of Enterobacter hormaechei phage was successfully isolated and identified and named H2.The phage formed uniform, round and transparent plaque on the double-layer agar plate.The results of transmission electron microscopy showed that the head of phage H2 was icosahedral symmetry, with a diameter of 85 nm±2 nm and a long tail with a tail filament length of 190 nm±2 nm, was a member of Caudovirales and the Mycobacteriophages family.The results of temperature and pH stability of phage H2 showed that the titer remained stable in the range of 35-55 ℃ and pH 5.0-9.0.The best MOI was 0.0001.One-step growth curve according to the best MOI was drew, and it was measured that incubation period was about 20 min, the outbreak period was about 50 min, and the lysis amount was about 68 PFU/cell.【Conclusion】 Phage H2 had strong cracking ability, thermal stability and acid-base stability, which provided a good material for the research and development of subsequent phage preparations.

Key words: phage; isolation and identification; Enterobacter hormaechei; biological characteristics

中图分类号:

阳亭亭, 冯子娟, 黄轶, 周虹宏. 霍氏肠杆菌噬菌体的分离鉴定及生物学特性研究[J]. 中国畜牧兽医, 2024, 51(1): 323-329.

YANG Tingting, FENG Zijuan, HUANG Yi, ZHOU Honghong. Isolation, Identification and Biological Characteristics of Enterobacter hormaechei Phage[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 323-329.

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