食品科学 ›› 2019, Vol. 40 ›› Issue (10): 141-148.doi: 10.7506/spkx1002-6630-20180621-405

• 生物工程 • 上一篇    下一篇

副溶血性弧菌广谱裂解性噬菌体的筛选及其在海产品安全控制中的应用

郑小双1,高 璐1,2,张 辉3,饶胜其1,2,杨振泉1,2,*   

  1. 1.扬州大学食品科学与工程学院,江苏 扬州 225009;2.扬州大学 江苏省人兽共患病学重点实验室,江苏 扬州 225009;3.江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地,江苏省农业科学院农产品质量安全与营养研究所,江苏 南京 210014
  • 出版日期:2019-05-25 发布日期:2019-05-31
  • 基金资助:
    江苏省农业科技自主创新资金项目(CX(16)1028);江苏省人兽共患病重点实验室开放课题(R1508);扬州大学科技创新团队培养对象资助项目(2016)

Screening for Broad-Spectrum Lytic Phage against Vibrio parahaemolyticus and Its Application to Marine Products

ZHENG Xiaoshuang1, GAO Lu1,2, ZHANG Hui3, RAO Shengqi1,2, YANG Zhenquan1,2,*   

  1. 1. College of Food Science and Engineering, Yangzhou University, Yangzhou 225009, China; 2. Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China; 3. Key Laboratory for Food Quality and Safety of Jiangsu Province-State Key Laboratory Cultivation Base by Province and Ministry of Science and Technology, Institute of Food Safe and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
  • Online:2019-05-25 Published:2019-05-31

摘要: 以致病性副溶血性弧菌(Vibrio parahaemolyticus,Vp)菌株作为宿主,从污水样品中分离裂解性噬菌体,并对其生物学特性及其在模拟污染黄鱼中Vp的抑菌作用进行研究。结果获得5 株广谱裂解性噬菌体,形态鉴定属于肌尾噬菌体科(Myoviridae)、长尾噬菌体科(Siphoviridae)和盖噬菌体科(Corticoviridae),裂解谱结果显示单噬菌体分别可以对5~24 株不同来源的Vp菌株产生裂解效应,由5 株噬菌体构成的混合物VppMIX能裂解所有Vp菌株(42 株)。噬菌体分离株在60 ℃和pH 4~10条件下具有良好的裂解活性。单噬菌体不敏感突变频率(bacteriophage insensitive mutant frequency,BIMF)在10-3~10-5范围内,但VppMIX能够显著降低的BIMF值达到10-6。应用Vp模拟污染黄鱼片作为模型研究VppMIX的抑菌效果,结果显示在25 ℃恒温保藏12 h后,不同剂量VppMIX处理的黄鱼样品中Vp载量比对照组(未处理)降低了1.41~4.98(lg(CFU/g))(P<0.01)。本研究为海产品中致病性Vp的控制提供了新型生物抑菌剂。

关键词: 副溶血性弧菌, 噬菌体, 广谱, 抑菌作用

Abstract: Five lytic bacteriophages were isolated from sewage samples using pathogenic Vibrio parahaemolyticus (Vp) strains as hosts. The biological and antibacterial properties of the isolated bacteriophages were studied using artificially contaminated yellow croaker as a model. Transmission electron microscopy (TEM) revealed that the 5 phages belonged to the Myoviridae, Siphoviridae or Corticoviridae families. The lytic spectrum showed that the single phages could lyse 5–24 Vp strains, while their mixture (VppMIX) could lyse all 42 tested Vp strains. The activity of the phages remained stable at 60 ℃ and pH 4–10. The bacteriophage insensitive mutant frequency (BIMF) of the single phages was 10-3–10-5, while that of VppMIX was reduced to 10-6. The bacteriostatic test showed that number of Vp in artificially contaminated yellow croaker was significantly reduced by 1.41–4.98 (lg(CFU/g)) as compared with the control (P < 0.01) when treated by VppMIX at 25 ℃ for 12 h . This study provides a new biological bacteriostatic agent to control pathogenic Vp in marine products.

Key words: Vibrio parahaemolyticus, phage, broad-spectrum, bacteriostatic effect

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