一株金黄色葡萄球菌烈性噬菌体的生物学特性及其裂解效果

摘要/Abstract

摘要： 目的：以耐甲氧西林金黄色葡萄球菌为宿主菌进行噬菌体分离，并研究其生理特性和初步运用效果。方法：用双层平板法对噬菌体进行分离纯化，透射电镜观察其结构，进一步测定其裂解谱，研究其生理特性，评价其对细菌生物膜的清除作用，并以牛奶和牛肉为模型探究其抑菌效果。结果：分离到一株烈性噬菌体P65，可裂解耐甲氧西林金黄色葡萄球菌和其他金黄色葡萄球菌。透射电镜表明，噬菌体的头部呈多面体结构，整体长度为（236±5）nm、头部直径为（71±2）nm、尾部直径长（165±5）nm，属于有尾噬菌体目中的长尾噬菌体科。噬菌体的最佳感染复数为MOI=0.01，一步生长曲线显示，噬菌体的潜伏期为10 min，裂解周期为90 min，裂解量为35.46 PFU/cell。在pH 4-10和温度30-45 ℃条件下具有稳定的裂解活性。紫外照射70 min后，噬菌体的效价由1.91x108 PFU/mL降至2.65x104 PFU/mL。经噬菌体处理48 h后，菌株MRSA 2和MRSA 24的生物被膜清除率分别为91.3%和92.2%。在25℃条件下，P65在牛奶和牛肉细菌污染模型中有一定的抑菌效果。结论：分离到一株烈性噬菌体，该噬菌体在清除生物被膜和食品抗菌方面有一定的运用潜能。

关键词: 烈性噬菌体, 金黄色葡萄球菌, 生物学特性, 应用

Abstract: Objective: To isolate phage using methicillin-resistant Staphylococcus aureus (MRSA) as the host bacteria, and to study its physiological characteristics and preliminary application effects. Methods: The bacteriophage was separated and purified by the double-layer plate method. The structure was observed by transmission electron microscope; the cleavage profile was further measured; the physiological characteristics were studied; the biofilm removal effect was evaluated; and the antibacterial effect was explored by using milk and beef as models. Results: A potent bacteriophage P65 was isolated, which could lyse MRSA and other S. aureus. Transmission electron microscopy showed that the head of the phage was a polyhedral structure with an overall length of (236±5) nm, a head diameter of (71±2) nm, and a tail diameter of (165±5) nm. It belongs to Siphoviridae, long-tailed bacteriophage family. The optimal multiplicity of infection of the phage was MOI=0.01. The one-step growth curve showed that the incubation period of the phage was 10 min, the lysis period was 90 min, and the lysis amount was 35.46 PFU/cell. It had stable cleavage activity under the conditions of pH 4-10 and temperature 30-45 ℃. After 70 min of UV irradiation, the titer of the phage decreased from 1.91x108 PFU/mL to 2.65x104 PFU/mL. After 48 hours of phage treatment, the biofilm clearance rates of strains MRSA 2 and MRSA 24 were 91.3% and 92.2%, respectively. At 25°C, P65 had a certain antibacterial effect in milk and beef contamination models. Conclusion: A potent lytic phage was isolated, which would have certain potential for application in removing biofilms and food antibacterial.

Key words: Lytic phage, Staphylococcus aureus, biological characterization, application

中图分类号:

TS251.1

侯忠余李传友朱成林于基成唐俊妮. 一株金黄色葡萄球菌烈性噬菌体的生物学特性及其裂解效果[J]. 食品科学, 0, (): 0-0.

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