2株海水鱼源潜在益生菌的分离、鉴定及特性分析  

Isolation,Identification,and Characterization of Two Potential Probiotics from Marine Fish

作　　者：杜文勇 王腾腾 韩慧宗 王斐 张明亮 宋元照 姜海滨 DU Wenyong;WANG Tengteng;HAN Huizong;WANG Fei;ZHANG Mingliang;SONG Yuanzhao;JIANG Haibin(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai201306,China;Shandong Marine Resource and Environment Research Institute,Yantai,264006,China)

机构地区：[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,山东烟台264006

出　　处：《渔业科学进展》2023年第3期188-199,共12页Progress in Fishery Sciences

基　　金：财政部和农业农村部:国家现代农业产业技术体系专项;山东省自然科学基金青年项目(ZR2020QC202);烟台市科技创新发展计划项目(2020MSGY073,2021XDHZ054)共同资助。

摘　　要：为筛选用于微生态制剂研发的海水鱼源益生菌,本研究对海捕野生许氏平鲉(Sebastes schlegelii)和大泷六线鱼(Hexagrammos otakii)的消化道内壁黏膜样品进行细菌分离纯化,获得80株可培养细菌。使用选择性培养基对菌株的产酶能力进行测定,选取海水鱼常见病原菌为指示菌测定分离菌株产物的抑菌活性,筛选出2株潜在益生菌TS2和TH8,并进行菌株的生理生化检测、16S rDNA序列分析、生长特性及其对宿主安全性的研究。结果显示,TS2产蛋白酶(protease)、淀粉酶(amylase)和脂肪酶(lipase),其无菌培养产物可显著抑制鳗弧菌(Vibrio anguillarum)、副溶血弧菌(Vibrio parahaemolyticus)、哈维氏弧菌(Vibrio harvey)和假交替单胞菌(Pseudoalteromonas nigrifaciens)的生长。TH8产蛋白酶和脂肪酶,其无菌培养产物可显著抑制鳗弧菌、溶藻弧菌(Vibrio alginolyticus)、副溶血弧菌、假交替单胞菌、嗜水气单胞菌(Aeromonas hydrophila)、金黄色葡萄球菌(Staphyloccocus aureus)和大肠杆菌(Escherichia coli)的生长。基于细菌生理生化检测和16S rDNA序列比对,鉴定TS2为枯草芽孢杆菌(Bacillus subtilis),TH8为河流漫游球菌(Vagococcus fluvialis)。TS2在温度为15~40℃、Na Cl浓度为0~0.08 g/L、p H为5~9时生长较快,6 h进入对数期,26 h后进入稳定期;TH8在温度为20~40℃、Na Cl浓度为0~0.08 g/L、p H为5~12时生长较快,2 h进入对数期,14 h后进入稳定期。菌株对同源宿主的安全性检测发现,TS2和TH8在108 CFU/mL浓度条件下对同源宿主是相对安全的。筛选的枯草芽孢杆菌TS2、河流漫游球菌TH8产酶能力强,其产物能抑制多种病原菌的生长,具有广温、广盐、耐酸碱、生长速度快等优点,可作为候选菌株开发为微生态制剂,研究结果为其在海水鱼养殖业中的应用提供数据支撑。Marine fish are rich in minerals(calcium and iron),vitamins(riboflavin and niacin),nutrients that are beneficial for brain(fatty acids),docosahexaenoic acid that prevent cardiovascular and cerebrovascular diseases,and are an important source of protein for humans.In recent years,the nexus between the increased demand of high-quality protein and the decrease in marine fishery resources resulted in the intensification of marine farming.However,this high-density farming has led to various bacterial diseases frequently due to lack of disease control methods,which has caused economic losses in the farming industry and impeded the healthy development of the industry.Therefore,it is particularly important to reduce the rates of bacterial diseases and improve the survival rate of cultured fish.It is of particular concern on how to maintain intestinal health of cultured fish under this farming method.Probiotics,a kind of living microorganisms,are beneficial to the health of the host.In the 1980s,probiotics have been viewed as an environmentally friendly and effective product in aquaculture;they improve the host health by influencing intestinal microbiota and nonspecific immunity to increase disease resistance ability.Moreover,they can be used as an ideal substitute for antibiotics in aquaculture production.Probiotics from Bacillus,lactic acid bacteria,saccharomyces,and nitrobacteria are diverse,and the function of each varies widely.As the typical representative of probiotics,lactic acid bacteria,may inhibit the growth of pathogens by the production of its metabolites,such as lactic acid,acetic acid,peroxide hydrogen,and bacteriocin.Lactic acid bacteria are also able to balance intestinal microecological imbalance and maintain intestinal microbiota balance when the intestinal microbiota contains pathogenic bacteria or the host is treated with antibiotics.Bacillus is an aerobic or facultative anaerobic gram-positive bacteria,which is stable,possesses strong stress resistance and high resurrection rate,and can produce vario

关 键 词：许氏平鲉 大泷六线鱼 枯草芽孢杆菌 河流漫游球菌 产酶 抑菌 

分 类 号：S917.1[农业科学—水产科学]