| 养殖大黄鱼一种黏孢子虫病的组织病理学及检测方法初探 |
| |
| 引用本文： | 施慧,陈卓,丁慧昕,谢建军,汪玮,王庚申,何杰,许文军.养殖大黄鱼一种黏孢子虫病的组织病理学及检测方法初探[J].中国水产科学,2019,26(1):203-213. |
| |
| 作者姓名： | 施慧 陈卓 丁慧昕 谢建军 汪玮 王庚申 何杰 许文军 |
| |
| 作者单位： | 1. 浙江省海洋水产研究所, 浙江省海水增养殖重点实验室, 浙江 舟山 316021;2. 浙江海洋大学海洋与渔业研究所, 浙江 舟山 316021 |
| |
| 基金项目： | 浙江省自然科学基金项目（LY15C190008）；浙江省科技厅院所专项（2017F50016，2017F30039）；浙江海洋大学“水产”省一流学科开放课题（20160008）. |
| |
| 摘 要： | 运用组织病理切片、电镜观察及PCR扩增等方法对2017年在舟山采集到的临床表现"白鳃"症状的发病大黄鱼(Larimichthyscrocea)开展了病原学及快速检测方法的初步研究。组织病理观察显示,患病鱼的肝、脾、肾等内脏组织发生严重的病理变化,尤其是组织内红细胞发生明显的退行性变化,同时血液中红细胞数量显著减少。在病鱼组织的电镜超薄切片中可观察到直径约300～600 nm的孢子虫样结构。提取病鱼内脏组织总基因组DNA样本,采用1对针对寄生原虫的通用引物进行SSUrDNA的PCR扩增,最终获得大小为1471bp的特异性条带,经测序比对发现,该条带与GenBank中1种黏孢子虫Sinuolinea sp.的序列同源性最高,达89%。根据获得的SSU rDNA序列,建立了适用于该病临床检测的巢式PCR方法,最小灵敏度可达0.5 pg。研究表明,引起此次网箱养殖大黄鱼"白鳃"症状并导致鱼类大量死亡的是一类寄生性黏孢子虫。
|
| 关 键 词： | 大黄鱼 白鳃病 组织病理学 电镜 亚单位核糖体核糖核酸基因 黏孢子虫 巢式PCR |
| 修稿时间： | 2019/1/28 0:00:00 |
Preliminary study on the histopathology and detection methods of a Myxosporea parasite that causes white-gill disease in cultured Larimichthys crocea |
| |
| SHI Hui,CHEN Zhuo,DING Huixing,XIE Jianjun,WANG Wei,WANG Gengshen,HE Jie,XU Wenjun.Preliminary study on the histopathology and detection methods of a Myxosporea parasite that causes white-gill disease in cultured Larimichthys crocea[J].Journal of Fishery Sciences of China,2019,26(1):203-213. |
| |
| Authors: | SHI Hui CHEN Zhuo DING Huixing XIE Jianjun WANG Wei WANG Gengshen HE Jie XU Wenjun |
| |
| Institution: | 1. Marine and Fisheries Research Institute of Zhejiang Province, Key Laboratary of Marine Culture and Enhancement of Zhejiang Province, Zhoushan 316021, China;2. Marine and Fisheries Research Institute of Zhejiang Ocean University, Zhoushan 316021, China |
| |
| Abstract: | White-gill disease emerged among cage-cultured Larimichthys crocea in Zhoushan in 2017. Systematic etiological methods such as polymerase chain reaction, histopathological sections, and electron microscopy in sick fish with clinical symptoms were performed to characterize the pathogen. Histopathological results showed that tissue necrosis was present in the spleen, liver, and kidney of sick fish. Moreover, the erythrocytes in the tissues underwent obvious degenerative changes, and a decreased number of red blood cells was observed in the blood. Ultrathin sections used in the electron microscopy showed the presence of microspore-like shapes with diameters from 300 to 600 nanometre. Total DNA extracted from the pathological tissue was used to amplify the 18S ribosomal RNA gene (SSU rDNA). One specific gene was identified to be homologous to that in Sinuolinea sp. with 89% amino acid sequence homology. Our study results showed that the recent pandemic of white-gill disease that affected Pseudosciaena crocea in 2017, which was caused by Myxosporea, was different from the previous white-gill disease. A specific and sensitive detection of the pathogen was developed by using nested PCR, which only requires a minimum amount of 0.5 pg of template DNA and produces 104 more copies than the number of copies produced in single PCR amplifications. |
| |
| Keywords: | Larimichthys crocea white-gill disease histopathology electron microscopy SSU rDNA Myxosporea nested PCR |
| 本文献已被 CNKI 维普 等数据库收录！ |
| 点击此处可从《中国水产科学》浏览原始摘要信息 |
| 点击此处可从《中国水产科学》下载免费的PDF全文 |
|
