导　　师： 祁自忠;韩茵

学科专业： 0852

授予学位： 硕士

作　　者： ;

机构地区： 中国海洋大学

摘　　要： 仿刺参/(却japonicm/)养殖以其单品种产值最高的优势及每年20/%的增长速度，已逐步成为一项新的热点国民经济产业。然而，近年来随着仿刺参养殖规模的不断扩大，养殖环境日益恶化，病害频发，严重威胁该产业的健康、可持续发展。传统的抗生素和消毒剂的使用取得了一定成效，但是过量且频繁地使用导致耐药性菌株滋生，环境污染及食品安全等问题。有益菌以其能够抑制病原菌，调节动物肠道微生态环境，增强养殖动物免疫力，且绿色环保的优势逐步被认可并广泛应用。本研究将采用高通量16S rDNA序列分析技术研究患病仿刺参肠道和添加在词料中海泥的菌群结构组成，并结合传统的分离培养技术从仿刺参养殖区海泥中分离蹄选对仿刺参病原菌灿烂弧菌有显著括抗作用的潜在有益菌。同时从商品化的EM菌液中分离蹄选优势有益菌，并通过优化发酵条件，大规模应用于仿刺参的养殖生产。以上研究为有益菌在仿刺参养殖中的研发及应用提供理论和实践依据。所获得研究结果如下： 1.高通量16S rDNA序列分析技术发现患病仿刺参肠道细菌种类分布以变形菌门（Proteobacteira/)为主，占74.20/%；另有梭形菌门（Fusobacteria/)、拟杆菌门（Bacteroidetes/)、厚壁菌门（Firmicutes/)、放线菌门（Actinobacteira/)、蓝细菌门（Cyanobacteira/)、浮霉状菌门（Planctomycetes/)共7个门类。其中，蓝细菌门和浮霉状菌门的细菌仅检测到1株。进一步分析变形菌门细菌的组成，发现主要为Y-变形囷纲和S-变形囷纲的细囷，其分布分别占71.83/%和26.98/%。丫-变形菌纲中包括弧菌属（Vibrio/)、亮发菌属（LeucotMx/)、假单胞菌属/(Pseudomonas/)、希瓦氏菌属（Shewamlla/)、科威尔氏菌属（Colwellia/)、默里特氏菌属/(^MorUella/)等共22个属的细菌。其中弧菌在y-变形菌纲中的比例最高，占64/%；主要为� The culture of Apostichopus japonicus has become a new hot spot of the nationaleconomic industry with the highest production value and20/%production growth rateeach year. However, the expansion of the Apostichopus japonicus aquaculture led tothe deteirorative environment and frequent outburst of disease, which have been be aseirous threat for the sustainable development of sea cucumber /(Apostichopusjaponicus/) culture in China. Traditional use of antibiotics and disinfectants haveachieved some successes, but excessive and frequent use of them lead to a lot ofproblems, such as the emergence of drug resistance strains, environment pollution andfood safety. Probiotics has gradually been accepted and widely used in aquaculturewith its advantages of the antagonism to pathogenic bacteria, the intestinal bacterialbalance of animals, the immune enhancement of farmed animals, and so on. In thisstudy, Roche GS FLX System was used to study the diversity of the intestinalbacteiral community of diseased Apostichopus japonicus and sea mud in the culturalarea. The potential probiotics which have significant antagonism to Vibrio splendiduswere isolated from the sea mud. At the same time, a probiotics DLOl isolated from thecommercialized EM was optimized to the fermentation and applied in the cultivationof Apostichopus japonicus in large scale. The results provide theoretical basis for thedevelopment and application of probiotics in the cultivation of Apostichopusjaponicus. The results obtained are as follows: 1. Using FLX16S rDNA sequence analysis, diversity of the intestinal bacteiralcommunity of diseased Apostichopus japonicus are mainly Proteobacteira whichaccount for74.20/%. There are also other seven phylums, such as Fusobacteria,Bacteroidetes, Firmicutes, Actinobacteira, Cyanobacteira, Planctomycetes. Only onestrain is detected from Cyanobacteria and Planctomycetes. Analysis of thecomposition of Proteobacteria bacteria shows y-Proteobacteria and s-Proteobacteira account for71.83/%and26.98/%respectively, y-proteobactei

关 键 词： 仿刺参 肠道菌群 序列分析技术 有益菌 芽孢杆菌

领　　域： [农业科学] [农业科学]