食品科学 ›› 2017, Vol. 38 ›› Issue (8): 96-101.doi: 10.7506/spkx1002-6630-201708016

• 生物工程 • 上一篇    下一篇

海水鱼肠道源性抗哈维氏弧菌乳酸菌的筛选与鉴定

马欢欢,吕欣然,林 洋,白凤翎,励建荣   

  1. 1.渤海大学食品科学与工程学院,辽宁省食品安全重点实验室,生鲜农产品贮藏加工及安全控制技术国家地方联合 工程研究中心,辽宁 锦州 121013;2.北京林业大学生物科学与技术学院,北京 100083
  • 出版日期:2017-04-25 发布日期:2017-04-24
  • 基金资助:
    辽宁省科技厅攻关项目(2015103020);泰山学者蓝色产业领军人才团队支撑计划项目(鲁政办字(2015)19号)

Screening and Identification of Anti-Vibrio harveyi Lactic Acid Bacteria Derived from Marine Fish Intestine

MA Huanhuan, Lü Xinran, LIN Yang, BAI Fengling, LI Jianrong   

  1. 1. Food Safety Key Laboratory of Liaoning Province, National & Local Joint Engineering Research Center of Storage, Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products, College of Food Science and Technology, Bohai University, Jinzhou 121013, China; 2. College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China
  • Online:2017-04-25 Published:2017-04-24

摘要: 哈维氏弧菌是引起对虾和鱼弧菌病的主要病原菌,给海产养殖业造成重大的经济损失。利用1.0% CaCO3MRS琼脂从鲈鱼、刀鱼、牙鲆和大菱鲆等海水鱼肠道中分离到49 株乳酸菌,采用牛津杯琼脂扩散法从中筛选出对哈维氏弧菌具有较强拮抗作用的菌株YP4-5,抑菌直径为18.26 mm。通过蛋白酶、pH值和温度等因素对菌株YP4-5的无细胞上清液(cell free supernatant,CFS)抑菌物质进行初步分析,表明抑菌活性物质对蛋白酶敏感,pH2.5~4.5时对哈维氏弧菌具有抑制作用,具有良好的热稳定性,初步判断YP4-5 CFS中抑菌活性物质为细菌素类。0.4倍最小抑菌浓度YP4-5 CFS可有效抑制74.74%哈维氏弧菌的生长,扫描电子显微镜观察结果表明YP4-5 CFS使哈维氏弧菌细胞完整性破坏,细胞膜溶解,胞内物质外泄。经生理生化和16S rRNA鉴定菌株YP4-5为清酒乳杆菌,为预防和控制水产养殖中哈维氏弧菌感染研发微生态制剂提供良好的菌源。

关键词: 海水鱼肠道, 乳酸菌, 拮抗, 哈维氏弧菌, 筛选与鉴定

Abstract: Vibrio harveyi is a main pathogenic bacterium that can induce vibriosis of shrimp and fish in the marine environment, causing huge economic losses in aquaculture. In this paper, 49 strains of lactic acid bacteria (LAB) were isolated from the marine fish intestine by the use of 1.0% CaCO3 MRS agar, including seabass, coilia ectenes, Paralichthys lethostigma and turbot. strain YP4-5 isolated from the intestine of Paralichthys lethostigma had strong antagonistic activity against V. harveyi using the method of Oxford cup agar diffusion, showing an inhibitory zone diameter of 18.26 mm. Antimicrobial substances of the cell-free supernatant (CFS) produced by strain YP4-5 were sensitive to protease, heat stable and effective within the pH range of 2.5–4.5, and they were preliminary determined as bacteriocins. The growth curve indicated that 74.74% of V. harveyi was inhibited by treatment with the CFS at a concentration of 0.4 MIC. Furthermore, cell integrity damage, cell membrane dissolution and intracellular component leakage were observed by scanning electron microscope. Strain YP4-5 was identified as Lactobacillus sakei according to physiological and biochemical characteristics and 16S rRNA sequence analysis. This research can provide a good source of probiotics to prevent and control V. harveyi infection in aquaculture.

Key words: marine fish intestine, lactic acid bacteria, inhibition, Vibrio harveyi, screening and identification

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