Isolation and identification of the pathogen causing granulomas in internal organs of turbot <i>Scophthalmus maximus</i>

Zhongping Luo; Zhenguo Zhang; Shuang Hao; Xiaohui Bai; Wen‐Ping Jia; Qian Gao; Shouming Feng

doi:10.3724/sp.j.1118.2019.18277

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PDF HTML阅读 XML下载导出引用引用提醒大菱鲆内脏结节病病原的分离与鉴定 DOI: 作者: 作者单位: 1. 天津市水产研究所, 天津 300221;2. 上海海洋大学水产与生命学院, 上海 201306;3. 天津市水生动物疫病预防控制中心, 天津 300221 作者简介: 罗璋(1982-),男,博士,高级工程师,主要从事水产动物病害防治研究.E-mail:luozhang2010@163.com 通讯作者: 中图分类号: 941 基金项目: 天津市自然科学基金项目（16JCYBJC30000）；天津市水产产业技术体系创新团队项目（ITTFR2017008）；天津市农业科技成果转化与推广项目（201501030）；天津市科技计划项目（18ZXBFNC00030）。 Isolation and identification of the pathogen causing granulomas in internal organs of turbot Scophthalmus maximus Author: Affiliation: 1. Tianjin Fishery Research Institute, Tianjin 300221, China;2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;3. Tianjin Aquatic Animal Disease Prevention and Control Center, Tianjin 300221, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论摘要:2017年8月，天津市滨海新区某养殖场大菱鲆（）发生病害，累计死亡率约为25%，患病鱼体表无明显症状。通过肉眼和病理组织切片观察发现，患病鱼肾脏、脾脏、肝脏和肠道存在大量圆形结节，肾脏、肝脏和脾脏组织病变明显，肾小管坏死，肝细胞脂肪变性，脾脏中存在大量的坏死细胞。此外，在脾脏和肾脏组织中发现大量的抗酸杆菌。利用传统病原菌分离方法，从具有典型症状的濒死大菱鲆肾脏组织分离到优势菌株myco-10，利用该菌株注射攻毒能导致健康大菱鲆66.7%的死亡率，且表现出与自然发病鱼相同的症状。采用16S rDNA、基因序列分析，构建系统发育树并结合细菌形态特征、生理生化测试对菌株myco-10进行鉴定，结果显示，菌株myco-10的16S rDNA、Mycobacterium marinum）和溃疡分枝杆菌（）聚为一簇，生理生化反应与海分枝杆菌一致。综合生理生化特征和基因序列分析结果，将菌株myco-10鉴定为海分枝杆菌。这是中国首例分枝杆菌引起大菱鲆病害的报道，可为大菱鲆内脏结节病的防治提供参考资料。 Abstract:Turbot () is an economically important marine fish species cultured in China. However, disease is a major problem in the turbot aquaculture fishery, causing high mortality. In August 2017, a disease outbreak among turbot in a marine farm in Tianjin led to a cumulative mortality rate of 25%. The main symptom of the disease was abundant white granulomas in the internal organs of the turbot. To define the causative pathogens, bacteria were isolated from the spleen and kidneys of diseased fish using conventional methods. An artificial infection experiment was conducted to determine the pathogenicity of the isolated strain. The 16S rDNA, gene sequences, as well as morphological and biochemical characteristics of the strain, were examined. In addition, histopathological analysis was performed on the diseased turbot. Abundant white granulomas were observed in the spleen, kidneys, livers, and intestinal tracts of diseased fish. The hepatocytes in the livers of the diseased fish were damaged, and several irregular vacuolations were observed. Some of the spleen cells were enlarged and necrotic, and epithelial cells from the renal tubule showed vacuolar degeneration and necrosis. Furthermore, in the center of the pregranulomas, as well as kidney and spleen macrophages contained several phagocytized red, rod-shaped acid-fast bacteria. Strain myco-10 was isolated from spleens of diseased fish. Healthy turbot were subjected to bacterial challenge tests by intraperitoneal injection of 3×108cfu/fish of the isolated myco-10. The accumulative mortality rate was 66.7% within 30 days post-infection. Diseased turbot displayed symptoms similar to those observed in naturally infected individuals. Strain myco-10 showed positive pigmentation production after exposure to visible light, Tween-80 hydrolysis, urease, catalase at 68℃, and arylsulfatase, and negative results in terms of nitrate reductase and growth on 5% NaCl. Phylogenetic trees were constructed by matching 16S rDNA, spp. registered in GenBank. The results showed that strain myco-10 clustered with . Strain myco-10 was determined to be based on bacterial morphology, analytical profile index identification, and 16S rDNA, genes sequences. The results of this study suggest that is the causal agent of granulomas in the internal organs of turbot. To the best of our knowledge, this is the first report on spp. causing disease in turbot in China. Our findings will be useful as a reference for the prevention and control of mycobacteriosis in cultured turbot. 参考文献相似文献引证文献

Isolation and identification of the pathogen causing granulomas in internal organs of turbot <i>Scophthalmus maximus</i>

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