文章摘要
发病鲆鲽类分离菌株的16S rRNA基因测序分析
Sequencing and phylogenetic analysis of the 16S rRNA genes of bacterial strains isolated from diseased flatfish
投稿时间:2019-01-28  修订日期:2019-03-19
DOI:
中文关键词: 鲆鲽类,病原菌,16S rRNA基因,系统发育学分析,鉴定
英文关键词: flatfish  pathogen  16S rRNA gene  phylogenetic analysis  identification
基金项目:
作者单位E-mail
兰欣 中国科学院 海洋研究所 实验海洋生物学重点实验室
中国科学院大学 
250613345@qq.com 
李杰 中国水产科学研究院 黄海水产研究所  
李贵阳 中国水产科学研究院 黄海水产研究所  
肖鹏 中国科学院 海洋研究所 实验海洋生物学重点实验室  
莫照兰 中国水产科学研究院 黄海水产研究所 mozl@ysfri.ac.cn 
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中文摘要:
      细菌性疾病是我国海水养殖鲆鲽类的主要病害,为全面了解病原菌种类,本研究对1999年-2012年从山东、江苏、河北、天津等沿海地区养殖场发病鲆鲽鱼类中分离得到的124株优势菌株进行了16S rRNA基因测序和系统发育学分析。将基因序列与GenBank核酸序列数据库进行相似度比对分析,结果显示有83株与弧菌属(Vibrio sp.)细菌相似度最高,11株与气单胞菌属(Aeromonas sp.)细菌相似度最高,4株与爱德华氏菌属(Edwardsiella sp.)细菌相似度最高,26株为其他15种属的细菌。根据系统发育学分析结果,进一步将66株菌鉴定为16个种,优势种为溶藻弧菌(V. alginolyticus),哈氏弧菌(V. harveyi),鳗弧菌(V. anguillarum),杀鲑气单胞菌(A. salmonicida),迟缓爱德华氏菌(E. tarda)。选择其中的9株鳗弧菌和4株迟缓爱德华氏菌进行人工感染实验,结果显示其中7株鳗弧菌和3株迟缓爱德华氏菌对大菱鲆有较强的致病性。研究结果为阐明我国海水养殖鲆鲽类的流行病发生历史、病原种类、开展病原监测及疾病控制提供重要的参考。
英文摘要:
      Flatfish is reared as the major industrial aquaculture marine fish species in North China. In the culturing process, members of this species will be subject to the infection of a variety of pathogens. Diseases caused by bacterial pathogens is the major harm to flatfish culture in China. To investigate the bacterial pathogens in flatfish, we focused on 124 strains isolated from organs of diseased flatfish in Shandong, Jiangsu, Hebei, Tianjin etc. between 1999 and 2012. The 16S rRNA of isolates were sequenced, BLAST with GenBank, and phylogenetic analyzed using Mega 5.05. As a results, 66.90% isolates were identified as Vibrio (83 strains), 8.90% as Aeromonas (11 stains), 3.20% as Edwardsiella (4 strains), and other 15 genera (26 strains). According to the phylogenetic tree, 66 strains were identified as 16 species, with dominated species as V. alginolyticus, V. harveyi, V. anguillarum, V. damselae, V. furnissii, V. splendidus, A. allosaccharophila, A. salmonicida, E. tarda and other 7 species. To evaluate the pathogenicity of isolates, the virulence of V. anguillarum and E. tarda strains isolated from diseased fish were further determined by experimental infection based on 50% lethal dose (LD50) in turbot (Scophtalmus maximus). Results showed that 7 V. anguillarum strains and 3 E. tarda strains were pathogenic to fish. The LD50 were 105.1 cfu/fish to 106.8 cfu/fish for pathogenic V. anguillarum and 103.4 cfu/fish to 104.1 cfu/fish for pathogenic E. tarda. There are also 2 strains of V. anguillarum and 1 strain of E. tarda showed low virulence to turbot, with the LD50 higher than 107 cfu/fish. The presented results provide significant data to ascertain the bacterial pathogens of flatfish, which is important for making strategy of epidemiology, monitor and control the bacterial diseases.
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