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一株许氏平鲉(Sebastes schlegelii)肠道乳酸菌的分离鉴定及特性分析
王腾腾1,2, 韦秀梅2, 常城1,2, 韩慧宗2, 姜海滨2
1.上海海洋大学水产与生命学院 上海 201306;2.山东省海洋资源与环境研究院山东省海洋生态修复重点实验室 烟台 264006
摘要:
为筛选具有广谱抑菌活性的乳酸菌用于微生态制剂开发,本研究自野生许氏平鲉肠道内共分离出35株细菌,从中筛选出1株乳酸菌YH1,其革兰氏染色呈阳性,且可发酵脱脂乳产生凝固、无气泡、呈酸性现象,基于形态学观察、生理生化和16S rDNA序列鉴定乳酸菌YH1为食窦魏斯氏菌。分析了酸碱度、温度和NaCl含量对食窦魏斯氏菌YH1生长的影响;选取金黄色葡萄球菌、嗜水气单胞菌、溶藻弧菌、副溶血弧菌、迟钝爱德华氏菌、鳗弧菌、假交替单胞菌和哈维氏弧菌等多种病原指示菌,以琼脂扩散法测定YH1培养12h产物的抑菌活性;并采用肌肉注射和腹腔注射两种方式对YH1的动物安全性进行检测。实验结果显示,食窦魏斯氏菌YH1适宜的生长条件为pH5.7-8.8、温度20-40℃,最适生长条件为pH7.2-8.4,温度35℃,在MRS培养基中添加NaCl会抑制YH1的生长;食窦魏斯氏菌YH1的无菌培养产物可显著抑制金黄色葡萄球菌、嗜水气单胞菌、溶藻弧菌、副溶血弧菌、迟钝爱德华氏菌、鳗弧菌、假交替单胞菌、哈维氏弧菌等病原指示菌的生长;食窦魏斯氏菌YH1在108CFU/mL及以下浓度对许氏平鲉是相对安全的。以上研究表明了食窦魏斯氏菌YH1具有广谱抑菌性和动物安全性,为其应用于水产养殖业奠定了理论基础。
关键词:  乳酸菌YH1  食窦魏斯氏菌  许氏平鲉  鉴定  特性
DOI:10.11693/hyhz20160700163
分类号:
基金项目:山东省良种工程项目“优质抗病速生鱼类良种选育”,2014-2017;国家现代农业产业技术体系项目,CARS-50-Z09号。
IDENTIFICATION AND CHARACTERISTICS OF A LACTIC ACID BACTERIA ASSOCIATED FROM INTESTINE OF BLACK ROCKFISH SEBASTES SCHLEGELII
WANG Teng-Teng1,2, WEI Xiu-Mei2, CHANG Cheng1,2, HAN Hui-Zong2, JIANG Hai-Bin2
1.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;2.Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Shandong Marine Resource and Environment Research Institute, Yantai 264006, China
Abstract:
To screen wide-spectrum bacteriostatic activity for probiotics application, a lactic acid bacteria strain Weissella cibaria YH1 was recognized among 35 bacterial strains isolated from the intestine of wild Sebastes schlegelii. Morphological observation, physiological and biochemical characteristic analyses, and 16S rDNA sequencing were applied. Skim milk was fermented into coagulation with this gram-positive YH1 without bubbling, and the supernatant was acidic. The effects of pH, temperature, and NaCl content on the growth of strain YH1 were analyzed. Some indicator pathogens (Staphylococcus aureus, Aeromonas hydrophila, Vibrio alginolyticus, Vibrio Parahaemolyticus, Edwardsiella tarda, Vibrio anguillarum, Pseudoalteromonas sp, and Vibrio harveyi) were used to measure the bacteriostatic activity of the strain product in agar diffusion method. Meanwhile, animal safety of strain YH1 was tested by intramuscular and intraperitoneal injections to S. schlegelii. Results show that YH1 can survive in range of pH 5.7-8.8 and 20-40℃, and best at 7.2-8.4 and 35℃. The growth of YH1 was inhibited by adding NaCl to MRS medium. The agar diffusion assay revealed that the product of YH1 inhibited the growth of all the indicator pathogens mentioned above. YH1 was safe to S. schlegelii as the survival rate post-injection reached 100%. Therefore, YH1 could inhibit the growth of wide-spectrum bacteria, and shall be developed for more applications in aquaculture.
Key words:  lactic acid bacteria strain YH1  Weissella cibaria  Sebastes schlegelii  identification  characterization
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