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| 从养殖贝类和大菱鲆分离的海洋发光弧菌的分类地位分析 |
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刘 斌1,2, 叶德赞1,2, Günter Jost3, 林 义1,2, 黄翔玲1,2, 郑森林1
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1.国家海洋局 第三海洋研究所;2.国家海洋局海洋 生物遗传资源重点实验室;3.德国莱布尼兹波罗的海研究所
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| 摘要: |
| 海洋发光弧菌是海水养殖中重要的条件致病菌。从福建省晋江市附近水产养殖场花蛤、牡蛎软组织及波罗地海大菱鲆肠道内含物中分离得到16 株发光细菌, 采用ARDRA(amplified ribosomal DNArestriction analysis)、16S rDNA 序列测定、Biolog 碳源代谢分析及药敏试验等方法对这些菌株的分类地位进行了分析。结果表明: 基于ARDRA 分析指纹图谱, 16 株发光菌分属于3 个OTUs(OperationalTaxonomic Unit, OTU); 对菌落和细胞形态、ARDRA 指纹图谱及来源不同的5 个菌株进行16S rDNA序列测定和系统发育分析, 结果显示这些菌株均属于Vibrio 属, 分别与Vibrio campbellii, V. orientalis, V.fischeri 及V. azureus 等亲缘关系最近, 表明ARDRA 方法存在一定的局限性, 难以区分同一属内的近缘种; Biolog 碳源代谢分析无法将海洋细菌鉴定至种的水平, 但可将亲缘关系较近的菌株划分至同一群体; 5 株菌的药敏试验结果显示各株菌均对复方新诺明和庆大霉素等敏感, 且亲缘关系较近的菌株也具有相似的药物敏感性。四种方法相结合可将细菌区分为属内分类地位相一致的不同群体, 为后续更精确的在生理生化方面鉴定细菌提供参考, 同时为海水养殖中防治潜在的由发光弧菌引发的病害提供有益的帮助。 |
| 关键词: 海洋发光弧菌 ARDRA 系统发育分析 Biolog 碳源代谢分析 药敏试验 |
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| 基金项目:中德合作项目(HB080302(1)); 海洋公益性创业科研专项(200905011) |
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| Classification of marine luminous bacteria isolated from farming seashells and Scophthatmus maximus |
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| Abstract: |
| Marine luminous vibrios are important opportunistic pathogens in marine aquaculture. 16 luminous bacteria were isolated from the soft bodies of clams (Ruditapes philippinarum) and of oysters (Saccostrea cucullata) collected in the aquacultural farms near Jinjiang in Fujian Province and from the gastroenteric contents of turbot fish (Scophthatmus maximus) from Baltic Sea. To ascertain the taxonomic statuses of the bacteria, four methods were applied. The ARDRA (amplified ribosomal DNA restriction analysis) fingerprints of 16 strains showed three different operational taxonomic units (OTUs). According to the bacterial morphology of colonies and cells, the ARDRA fingerprints, and sample sources, five strains were sent for sequencing of 16S rDNA. The results of phylogenetic analysis of 16S rDNA evinced that they are all belong to genus Vibrio, and most closely related to V. fischeri, V. orientalis, V. azureus, and V. campbellii respectively. This result indicates that ARDRA method has limitation when distinguishing different species that are closely related from the same genus. Biolog C-Source-Utilizing analysis could not identify marine bacteria to species level according to the database, but could classify the closely related bacteria species to the same group. The results of drug sensitive tests by the K-B method showed that the closely related species also had similar drug sensitivities. All of the five strains were sensitive to sulfamethoxazole and gentamicin, which will be helpful to treat potential luminous bacteria diseases in marine aquaculture. The combination of the four methods could classify the bacteria species to different groups and this could be the basis for further physiological characterization. |
| Key words: marine luminous bacteria ARDRA phylogenetic analysis biolog system drug sensitive test |