一株高效氨氮及亚硝态氮去除功能菌株的分离鉴定及在生物絮团对虾养殖中的应用  

Isolation and identification of an ammonia-N-and nitrite-N-degrading strain and its application in bioflocs for shrimp culture system

作　　者：丁雄祺 谢媚 陈偿[1,2,3] 管云雁[1,2] 方建相 云龙 DING Xiongqi;XIE Mei;CHEN Chang;GUAN Yunyan;FANG Jianxiang;YUN Long(Key Laboratory of Tropical Marine Bio-resources and Ecology,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China;Guangdong Provincial Key Laboratory of Applied Marine Biology,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China;Xisha/Nansha Ocean Observation and Research Station,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院南海海洋研究所,中国科学院热带海洋生物资源与生态重点实验室,广东广州510301 [2]中国科学院南海海洋研究所,广东省应用海洋生物学重点实验室,广东广州510301 [3]中国科学院南海海洋研究所西沙、南沙深海海洋环境观测研究站,广东广州510301 [4]中国科学院大学,北京100049

出　　处：《中国水产科学》2019年第5期959-970,共12页Journal of Fishery Sciences of China

基　　金：国基自然科学基金面上项目(Y8GM081001);广州市科技计划项目(Y8D1111001)

摘　　要：从凡纳滨对虾(Litopenaeus vannamei)养殖水体中分离出具有高效氨氮及亚硝态氮去除功能的菌株Y2,生理生化和16S rRNA基因序列比对分析结果显示该菌株为麦氏交替单胞菌(Alteromonas macleodii)。进一步通过生长实验进行温度、酸碱度、盐度的培养条件优化,利用抗生素药敏实验筛选菌株特定抗性;通过卤虫浸泡感染的方法检测麦氏交替单胞菌Y2的安全性,并测定海水培养液OD600及含氮无机污染物的浓度,探究菌株Y2生长与水体中氨氮、亚硝态氮、硝态氮之间的动态变化关系;通过28 d对虾养殖试验,监测水质、生物絮团形成量、致病菌数量及对虾成活率生长速率,进一步阐明菌株在实际养殖中的功效。结果表明,该菌株Y2对苯唑西林、克林霉素有抗性;对卤虫的48 h半致死浓度高于1.9×10^8 cfu/mL,显著高于哈氏弧菌(102 cfu/mL)。此外,该菌具有持续去除水体中氨氮、亚硝态氮的功能。在养殖实验中能抑制潜在病原菌弧菌生长、提高对虾存活率及生长率,并且能在水体中稳定存活较长时间。综上所述,菌株Y2是养殖用益生菌制剂的优良备选菌株,可作为生物絮团养殖系统中调节水质的关键菌株。In recent years,biofloc technology(BFT)has become an important water-saving and emission-reducing aquaculture model in aquaculture development.However,in high-density farming ponds,the concentrations of ammonia and nitrite are often much higher than established standards.High ammonia and nitrite concentrations are harmful in aquaculture,where they increase the stress experienced by shrimp,resulting in disease outbreaks.The removal of ammonia and nitrite is an effective method of reducing water pollution resulting from residual food and feces.The Y2 strain,which can efficiently remove ammonia and nitrite,was isolated from the culture water of Litopenaeus vannamei.Experiments investigating the physiology-biochemistry of this strain and the similarity of 16S rRNA gene sequences have been conducted.The safety of Y2 was evaluated by an immersion infection and antimicrobial susceptibility test.The fermentation conditions,including temperature,pH,and salinity,were optimized,and dynamic changes between bacterial growth and the concentration of pollutants in water were investigated.Water quality,the amount of bioflocs,the number of pathogenic bacteria,and the growth and survival rates of shrimps were monitored through a 28-day culture experiment.The Y2 strain was preliminarily identified as Alteromonas macleodii,based on physiological and biochemical characteristics and 16S rRNA sequence analysis.The strain was resistant to oxacillin and clindamycin.Optimum fermentation of the Y2 strain was observed under conditions of 30℃,pH 7.0,and 2%salinity.The 48 h half-lethal concentration of the strain to Artemia was higher than that of Vibrio harveyi.The LC50 of strain Y2 exceeded 10^8 cfu/mL,and this strain was nontoxic to cultured animals.The Y2 strain continuously removed ammonia and nitrite from aquaculture water.It inhibited the growth of vibrios,increased the survival and growth rates of shrimps,and was able to survive in water for a long period of time.Therefore,the Y2 strain has high potential as a probiotic for aquacult

关 键 词：氨氮 亚硝态氮 去除 麦氏交替单胞菌 凡纳滨对虾 菌株安全性 

分 类 号：S96[农业科学—水产养殖]