大口黑鲈肠道益生菌的分离与鉴定  

Isolation and Identification of Intestinal Probiotics from Micropterus salmoides

作　　者：赵浩航 康旭 黄子威 温依铭 汪志文[1,2,3] 喻大鹏 甘桢 夏洪丽 夏立群 鲁义善[1,2,3] ZHAO Hao-hang;KANG Xu;HUANG Zi-wei;WEN Yi-ming;WANG Zhi-wen;YU Da-pengGAN Zhen;XIA Hong-li;XIA Li-qun;LU Yi-shan(Fisheries College of Guangdong Ocean University,Zhanjiang 524088,China;Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy culture,Zhanjiang 524088,China;Shenzhen Institute of Guangdong Ocean University/Guangdong Provincial Engineering Research Center for Aquatic Animal Health Assessment/Shenzhen Public Service Platform for Evaluation of Marine Economic Animal Seedings,Shenzhen 518120,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东省水产动物病害防控与健康养殖重点实验室,广东湛江524088 [3]广东海洋大学深圳研究院/广东省水生动物健康评估工程技术研究中心/深圳市海水经济动物种苗健康评价公共技术服务平台,广东深圳518120

出　　处：《广东海洋大学学报》2023年第4期62-68,共7页Journal of Guangdong Ocean University

基　　金：深圳市可持续发展专项(KCXFZ20211020165547010);深圳市科技计划(JCYJ20220530140813030)。

摘　　要：【目的】分离鉴定大口黑鲈(Micropterus salmoides)肠道益生菌,评估分离菌用作饲料添加剂的益生潜力。【方法】采集大口黑鲈肠道样本,采用稀释涂布法分离细菌,经16S rRNA序列比对和生化试验鉴定种属,并进行抗逆性能、黏附能力、溶血活性和药敏特性等鉴定。【结果】从大口黑鲈肠道分离菌株84株,经测序、blast比对,筛选出3株同源性最高的菌株MS-1、MS-2、MS-3,分别鉴定为地衣芽孢杆菌(Bacillus licheniformis)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和贝莱斯芽孢杆菌(Bacillus velezensis)。3株分离菌均为γ-溶血,不具有生物膜,疏水性50%~93.3%,自凝集性43.6%~69.2%;可在pH 2.0~12.0环境和5.0 g/L胆盐中存活;经高温处理后仍可生长;3株菌对大多数抗生素敏感。【结论】芽孢杆菌MS-1、MS-2和MS-3抗逆性高,黏附性能及饲料学安全性高,可作为大口黑鲈养殖中潜在益生菌。【Objective】To isolate and identify intestinal probiotics of Micropterus salmoides,and evaluate the probiotics potential of the isolated bacteria as feed additives.【Method】Intestinal samples of Micropterus salmoides were collected and bacteria were isolated by dilution coating method,and species were determined by 16S rRNA sequence comparison screening and biochemical tests.Subsequently,stress resistance,adhesion,hemolytic activity and drug sensitivity characteristics were identified.【Result】Eighty-four strains of probiotics were isolated from the intestine of Micropterus salmoides.After sequencing and blast comparison,three strains of probiotics with the highest homology,MS-1,MS-2 and MS-3,were screened out and identified as Bacillus licheniformis,Bacillus amyloliquefaciens and Bacillus velezensis,respectively.The three isolates showedγ-hemolysis,without biofilm,with 50%‒93.3%hydrophobicity and 43.6%‒69.2%self-agglutination.They could survive in pH 2.0‒12.0 environment and 5.0g/L bile salt and grow after high temperature treatment and were sensitive to most antibiotics.【Conclusion】Bacillus MS-1,MS-2 and MS-3 have high stress resistance,adhesion and feed safety,and can be used as potential probiotics in the aquaculture of Micropterus salmoides.

关 键 词：大口黑鲈 肠道微生物 益生菌 分离与鉴定 芽孢杆菌 

分 类 号：S965.199[农业科学—水产养殖] Q93-3[农业科学—水产科学]