水产养殖用芽孢杆菌制剂的检测Detection of Bacillus Preparations Used for Aquaculture
温崇庆;薛明;张良军;李兴;吴勇;
摘要(Abstract)：
通过平板菌落计数法检测编号为GD1、GD2、GD3、GD4、GD5、JS、JX、SD1和SD2等9种水产养殖用芽孢杆菌制剂的有效活菌数,结合形态、生理生化和16S rRNA基因序列分析对各制剂中分离的产芽孢细菌代表菌株进行鉴定。结果显示:除GD1、GD2和GD5制剂未标明芽孢数量外,其余制剂检测到的芽孢数量均低于或远低于标注数量;经鉴定,各制剂中最优势类型,GD2、GD3、GD5、SD1和SD2制剂为地衣芽孢杆菌(Bacillus licheniformis),GD4、JS和JX制剂为解淀粉芽孢杆菌(B.amyloliquefaciens),GD1制剂为枯草芽孢杆菌(B.subtilis)。除GD4、GD5和SD1制剂未标明具体菌种以及GD2的标注菌种与检测结果不一致外,其他制剂的标注菌种与被鉴定的优势类型芽孢杆菌基本一致。
关键词(KeyWords)： 芽孢杆菌;制剂;数量;鉴定;水产养殖
基金项目(Foundation): 广东省海洋渔业科技推广专项(A200808A04)
作者(Authors): 温崇庆;薛明;张良军;李兴;吴勇;
参考文献(References)：
- [1]Hong H A,Duc L H,Cutting S M.The use of bacterial spore formers as probiotics[J].FEMS Microbiology Reviews,2005,29(4):813-835.
- [2]于明超,李卓佳,文国樑.芽孢杆菌在水产养殖应用中的研究进展[J].广东农业科学,2007(11):78-81.
- [3]温崇庆,薛明,张金燕,等.水产养殖用蛭弧菌类生物制剂的检测[J].水产学报,2009,33(2):326-333.
- [4]沈德龙,李俊,姜昕,等.GB 20287-2006农用微生物菌剂[S].北京:中国标准出版社,2006.
- [5]Koransky J R,Allen S D,Dowell V R.Use of ethanol for selective isolation of spore-forming organisms[J].Applied and environmental microbiolgy,1978,35(4):762-765.
- [6]温崇庆,赖心田,何红,等.两株对虾育苗用益生芽孢杆菌的筛选和鉴定[J].水生生物学报,2007,31(4):453-459.
- [7]Tamura K,Dudley J,Nei M,et al.MEGA4:molecular evolutionary genetics analysis(MEGA)software version 4.0[J].Molecular biology and evolution,2007,24:1596-1599.
- [8]Slepecky R A,Hemphill H E.The genus Bacillus—nonmedical[M].//Dworkin M.The Prokaryotes,Volume 4,3rd ed.New York:Springer,2006:534-535.
- [9]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:43-65;364-384.
- [10]Nakamura L K.Deoxyribonucleic acid relatedness of lactose-positive Bacillus subtilis strains and Bacillus amyloliquefaciens[J].Journal of bacteriology,1987,37(4):444-445.
- [11]Ahmed I,Yokota A,Fujiwara T.A novel highly boron tolerant bacterium,Bacillus boroniphilus sp.nov.,isolated from soil,that requires boron for its growth[J].Extremophiles,2007,11(2):217-224.
- [12]An S Y,Asahara M,Goto K,et al.Terribacillus saccharophilus gen.nov.,sp.nov.and halophilus sp.nov.,spore-forming bacteria isolated from field soil in Japan[J].International journal of systematic and evolutionary microbiology,2007,57(1):51-55.
- [13]Wang Y B,Li J R,Lin J.Probiotics in aquaculture:Challenges and outlook[J].Aquaculture,2008,281(1-4):1-4.
- [14]Orozco-Medina C,Maeda-Martínez A M,López-Cortés A.Effect of aerobic Gram-positive heterotrophic bacteria associated with Artemia franciscana cysts on the survival and development of its larvae[J].Aquaculture,2002,213(1-4):15-29.
- [15]Fjellheim A J,Klinkenberg G,Skjermo J,et al.Selection of candidate probionts by two different screening strategies from Atlantic cod(Gadus morhua L.)larvae[J].Veterinary microbiology,2010,144(1-2):153-159.
- [16]Priest F G,Goodfellow M,Shute A,et al.Bacillus amyloliquefaciens sp.nov.,nom.rev.[J].International journal of systematic bacteriology,1987,37(1):69-71.