Seasonal incidence of autochthonous antagonistic Roseobacter spp. and Vibrionaceae strains in a turbot larva (Scophthalmus maximus) rearing system
Seasonal incidence of autochthonous antagonistic Roseobacter spp. and Vibrionaceae strains in a turbot larva (Scophthalmus maximus) rearing system
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DOI：
10.1128/aem.70.12.7288-7294.2004
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发表时间：
2004-12-01
影响因子：
4.4
通讯作者：
Gram, L
中科院分区：
文献类型：
--
作者：
Hjelm, M;Riaza, A;Gram, L
经过一年的时间，从两个大菱鲆幼体饲养场中分离出了对鱼类幼体致病菌具有抑制作用的细菌。从饲养场所采集样品，例如水箱壁、水和幼虫饲料，并使用复制平板测定法分离出具有对抗鳗弧菌拮抗活性的细菌。从海洋琼脂平板上复制大约 19,000 个菌落，并从在鳗弧菌层中产生透明区的菌落中分离出 341 个菌株。在良好扩散琼脂测定中进行测试时，173 株菌株保留了对鳗弧菌和灿烂弧菌的抗菌活性。生化测试鉴定出 132 株菌株为玫瑰杆菌属 (Roseobacter spp)。 31 株为弧菌科菌株。对三个菌株的 16S rRNA 基因进行部分测序，证实其为加氏玫瑰杆菌。玫瑰杆菌属在春季和初夏的几个月里尤其孤立。 132 种玫瑰杆菌属的亚型。通过使用两个引物随机扩增多态性 DNA 来对菌株进行分析表明，这些菌株形成了一个非常同质的群体。因此，看来两个养鱼场都存在相同的亚型，并且在一年的调查期间持续存在。这表明该亚型有一个常见的、常规的来源，或者某种特定亚型可能已在养鱼场的某些区域建立起来。 31 种拮抗剂被鉴定为弧菌属，其中 18 种是鳗弧菌，但不是血清型 O1 或 O2。玫瑰杆菌属特别是从幼体池壁上分离出菌株，有可能在大菱鲆幼体的饲养环境中建立一种拮抗的、有益的微生物群落，从而限制病原菌的生存。
Bacteria inhibitory to fish larval pathogenic bacteria were isolated from two turbot larva rearing farms over a 1-year period. Samples were taken from the rearing site, e.g., tank walls, water, and feed for larvae, and bacteria with antagonistic activity against Vibrio anguillarum were isolated using a replica plating assay. Approximately 19,000 colonies were replica plated from marine agar plates, and 341 strains were isolated from colonies causing clearing zones in a layer of V. anguillarum. When tested in a well diffusion agar assay, 173 strains retained the antibacterial activity against V. anguillarum and Vibrio splendidus. Biochemical tests identified 132 strains as Roseobacter spp. and 31 as Vibrionaceae strains. Partial sequencing of the 16S rRNA gene of three strains confirmed the identification as Roseobacter gallaeciensis. Rosebacter spp. were especially isolated in the spring and early summer months. Subtyping of the 132 Roseobacter spp. strains by randomly amplified polymorphic DNA with two primers revealed that the strains formed a very homogeneous group. Hence, it appears that the same subtype was present at both fish farms and persisted during the 1-year survey. This indicates either a common, regular source of the subtype or the possibility that a particular subtype has established itself in some areas of the fish farm. Thirty-one antagonists were identified as Vibrio spp., and 18 of these were V. anguillarum but not serotype O1 or O2. Roseobacter spp. strains were, in particular, isolated from the larval tank walls, and it may be possible to establish an antagonistic, beneficial microflora in the rearing environment of turbot larvae and thereby limit survival of pathogenic bacteria.