赵浩航,康旭,黄子威,温依铭,汪志文,喻大鹏,甘桢,夏洪丽,夏立群,鲁义善

• 1:广东海洋大学水产学院
• 2:广东省水产动物病害防控与健康养殖重点实验室
• 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

摘要(Abstract)：

【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。

关键词(KeyWords)： 大口黑鲈;肠道微生物;益生菌;分离与鉴定;芽孢杆菌

Abstract:

Keywords:

基金项目(Foundation): 深圳市可持续发展专项（KCXFZ20211020165547010）;; 深圳市科技计划（JCYJ20220530140813030）

作者(Author): 赵浩航,康旭,黄子威,温依铭,汪志文,喻大鹏,甘桢,夏洪丽,夏立群,鲁义善

参考文献(References)：

• [1]卢俊桦,蔡春静,李璇,等.鼠李糖乳杆菌对加州鲈生理特性与肠道菌群的影响[J].饲料工业, 2022, 43(20):16-21.
• [2] ZHAO L L, TANG G, XIONG C, et al. Chronic chlorpyrifos exposure induces oxidative stress, apoptosis and immune dysfunction in largemouth bass(Micropterus salmoides)[J].Environmental Pollution, 2021, 282:117010.
• [3] ZHOU Y L, HE G L, JIN T, et al. High dietary starch impairs intestinal health and microbiota of largemouth bass,Micropterus salmoides[J]. Aquaculture, 2021, 534:736261.
• [4] TRAN N T, YANG W, NGUYEN X T, et al. Application of heat-killed probiotics in aquaculture[J]. Aquaculture, 2022,548:737700.
• [5] BANERJEE G, RAY A K. The advancement of probiotics research and its application in fish farming industries[J].Research in Veterinary Science, 2017, 115:66-77.
• [6]郭同旺,周书洪,郑守伟,等.几种常见水产养殖益生菌的应用研究现状及展望[J].盐科学与化工, 2020, 49(11):3-7.
• [7]张冬梅,颜浩骁,罗茂林,等.饲料中添加枯草芽孢杆菌对大口黑鲈幼鱼生长、肠道组织结构、抗氧化能力、免疫能力和肠炎的影响[J].动物营养学报, 2022, 34(1):575-588.
• [8]胡娟,高辰辰,药园园,等.饲用枯草芽孢杆菌HGcc-1对鲤肠肝健康、血清补体及肠道菌群的影响[J].水产学报,2021, 45(10):1753-1763.
• [9]张静,陈红莲,鲍俊杰,等.水产养殖中嗜水气单胞菌拮抗菌的研究进展[J].江苏农业科学, 2020, 48(17):21-33.
• [10]温俊,孙笑非.水产动物肠道微生态调控研究进展[J].饲料研究, 2009(9):68-70.
• [11] CHEESMAN S E, GUILLEMIN K. We know you are in there:Conversing with the indigenous gut microbiota[J].Research in Microbiology, 2007, 158(1):2-9.
• [12] GUPTA V, GARG R. Probiotics[J]. Indian Journal of Medical Microbiology, 2009, 27(3):202-209.
• [13] JE?EWSKA-FR?CKOWIAK J, SEROCZY?SKA K,BANASZCZYK J, et al. The promises and risks of probiotic Bacillus species[J]. Acta Biochimica Polonica, 2018, 65(4):509-519.
• [14]巨向红,谢晓杭,张金燕,等.一株纳豆芽孢杆菌的饲料学特性研究[J].广东海洋大学学报, 2015, 35(4):87-92.
• [15] GUO X, CHEN D D, PENG K S, et al. Identification and characterization of Bacillus subtilis from grass carp(Ctenopharynodon idellus)for use as probiotic additives in aquatic feed[J]. Fish&Shellfish Immunology, 2016, 52:74-84.
• [16] WANG Y P, LI A Y, JIANG X, et al. Probiotic potential of Leuconostoc pseudomesenteroides and Lactobacillus strains isolated from yaks[J]. Frontiers in Microbiology,2018, 9:2987.
• [17]孙承文,巩华,赖迎迢,等.水产源芽胞杆菌的分离鉴定及生物学功能分析[J].微生物学杂志, 2020, 40(6):30-35.
• [18] KIM S K, GUEVARRA R B, KIM Y T, et al. Role of probiotics in human gut microbiome-associated diseases[J].Journal of Microbiology and Biotechnology, 2019, 29(9):1335-1340.
• [19] ZENG Z, ZHANG J, LI Y, et al. Probiotic potential of Bacillus licheniformis and Bacillus pumilus isolated from tibetan yaks, China[J]. Probiotics and Antimicrobial Proteins,2022, 14(3):579-594.
• [20] KIM D H. Gut microbiota-mediated pharmacokinetics of ginseng saponins[J]. Journal of Ginseng Research, 2018,42(3):255-263.
• [21] LIU H T, WANG S F, CAI Y, et al. Dietary administration of Bacillus subtilis HAINUP40 enhances growth, digestive enzyme activities, innate immune responses and disease resistance of tilapia, Oreochromis niloticus[J]. Fish&Shellfish Immunology, 2017, 60:326-333.
• [22] ARGYRI A A, ZOUMPOPOULOU G, KARATZAS K A G, et al. Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests[J]. Food Microbiology,2013, 33(2):282-291.
• [23] NANDI A, DAN S K, BANERJEE G, et al. Probiotic potential of autochthonous bacteria isolated from the gastrointestinal tract of four freshwater teleosts[J]. Probiotics and Antimicrobial Proteins, 2017, 9(1):12-21.
• [24] RAMESH D, VINOTHKANNA A, RAI A K, et al. Isolation of potential probiotic Bacillus spp. and assessment of their subcellular components to induce immune responses in Labeo rohita against Aeromonas hydrophila[J]. Fish&Shellfish Immunology, 2015, 45(2):268-276.
• [25] CHRISTAKI E, MARCOU M, TOFARIDES A. Antimicrobial resistance in bacteria:mechanisms, evolution, and persistence[J]. Journal of Molecular Evolution, 2020, 88(1):26-40.
• [26] DONLAN R M. Biofilm formation:a clinically relevant microbiological process[J]. Clinical Infectious Diseases,2001, 33(8):1387-1392.