水产养殖用蛭弧菌类生物制剂的检测  

The detection of Bdellovibrio-and-like organisms in commercial preparations used for aquaculture

作　　者：温崇庆[1,2] 薛明[1] 张金燕 黄瑜[1] 周世宁[2] 

机构地区：[1]广东海洋大学水产学院,广东湛江524025 [2]中山大学生命科学学院有害生物控制与资源利用国家重点实验室,广东广州510275 [3]广东绿百多生物科技有限公司,广东湛江524022

出　　处：《水产学报》2009年第2期326-333,共8页Journal of Fisheries of China

基　　金：国家“八六三”高技术研究发展计划(2008AA09Z402)

摘　　要：通过双层平板法以不同细菌为宿主测定了8个厂家蛭弧菌类生物制剂的噬斑含量及噬斑分离物的宿主裂解范围;以3对靶向不同类群蛭弧菌类生物的特异引物对各产品及其噬斑分离物进行了PCR检测;对两株代表性蛭弧菌分离物的16S rRNA基因进行了测序,并通过构建蛭弧菌科进化树分析了它们的系统地位。结果表明,被检测的8个蛭弧菌类生物制剂中有6个质量明显不合格,仅两个样品被蛭弧菌科特异引物检测为阳性,被双层平板法证实有噬斑出现。其中一个样品JSF中存在两种形态和宿主范围不同的噬斑分离物,且不同宿主菌计数结果存在明显差异。16S rRNA基因序列分析表明这两种噬斑分离物JSF1和JSF2,分别属于蛭弧菌科的类群1(噬菌蛭弧菌)和未定种的类群5。但另一样品的噬斑分离物则被证实为非蛭弧菌类生物。Bdellovibrio-and-like organisms （BALOs） are predatory bacteria and have potential for biocontrol of pathogenic bacteria. In order to provide scientific reference for study, application and market supervision of the BALOs preparations, this study was to evaluate the quality of commercial BALOs preparations used for aquaculture. Using double layer agar technique, the plaques contents of BALOs preparations manufactured by eight companies was counted with different bacteria as preys, and the prey range of plaque isolates were tested. The preparations and the plaque isolates were also identified by PCR with three pairs of taxon-specific primers targeting various BALOs groups. The 16S rRNA genes of two representative Bdellovibrionaceae isolates were sequenced and compared with that of related sequences, and the phylogenetic tree of Bdellovibrionaceae was constructed based on genetic distance analysis. The results showed that most of the BALOs preparations being examined were not qualified, only two of the eight preparations gained positive amplification by Bdellovibrionaceae primers, and showed visible plaques on the double layer agar. One of the two Bdellovibrionaceae positive preparations, JSF, had two kinds of plaque isolates with different morphologies and prey patterns, and significant differences were found when counted with different preys. About 1.4 kb 16S rRNA genes were sequenced and deposited in the GenBank with accession numbers EU884925 and EU884926 for the two kinds of plaque isolates, JSF1 and JSF2, respectively. Based on the phylogenetic analysis of 16S rRNA gene sequences, JSF1 and JSF2 were offiliated with cluster I （ B. bacteriovorus ） and cluster 5 （ the species undefined ） respectively in Bdellovibrionaceae. However, the plaque isolates of the other preparation contained no BALOs.

关 键 词：水产养殖 蛭弧菌类生物 微生物制剂 检测 16S RRNA基因 

分 类 号：Q939.9[生物学—微生物学]