中国水产科学2023，Vol.30Issue(8)：1042-1053,12.DOI:10.12264/JFSC2023-0129
养殖三疣梭子蟹十足目虹彩病毒1的检出及组织病理学分析
Detection and histopathological analysis of Decapoda iridescent virus 1 in cultured Portunus trituberculatus
摘要
Abstract
In October 2022,an infectious disease outbreak was observed in farmed Portunus trituberculatus in Zhoushan,China.Samples of diseased crabs showing unspecific signs,such as anorexia,slow activity,and gill edema,were analyzed using parasitology,microbiology,histopathology,electron microscopy,and molecular identification.Parasitological and microbiological assays indicated that the diseased crabs were not infected with parasites or bacteria.The crab samples were tested by PCR methods recommended by World Organization for Animal Health and demonstrated to be free of Hematodinium,white spot syndrome virus(WSSV),mud crab discistrovirus-1(MCDV-1),and Scylla serrata reovirus(SSRV).Histopathological examination revealed eosinophilic inclusions in hematopoietic tissue and hemocytes in gills,hepatopancreas,and the myocardial tissue of diseased crabs.Meanwhile,transmission electron microscopy showed that the virus in diseased crab tissues exhibited a typical icosahedral structure with a mean diameter of 150 nm,which was similar to the morphological characteristics of Decapoda iridescent virus 1(DIV1).Nested PCR detection of DIV 1 result showed that the first step of the PCR produced a 457 bp amplicon and the second step of the PCR produced a 129 bp amplicon.Phylogenetic analyses using gene sequences of major capsid protein(MCP)and ATPase revealed amplified sequences,DS.MCP201010 and DS.ATP202210 sequences,had the highest homology with the MCP gene and ATPase gene of DIV 1,100％.Therefore,it was determined that the pathogen of the disease was DIV1.In addition,specific probes directed to the MCP and ATPase gene of DIV1 were designed to obtain the distribution of virus particles in the hepatopancreas,heart,muscles,and gills of diseased crabs by fluorescence in situ hybridization,which was consistent with the results of electron microscopy and nested PCR.This study presents the first report of DIV1 infection in cultured Portunus trituberculatus,which will provide a strong reference for the prevention and control of the disease.关键词
三疣梭子蟹/十足目虹彩病毒1/组织病理/原位杂交Key words
Portunus trituberculatus/Decapoda iridescent virus 1,DIV1/histopathology/in situ hybridization分类
农业科技引用本文复制引用
赵丹阳,施慧,许文军,何杰,王庚申..养殖三疣梭子蟹十足目虹彩病毒1的检出及组织病理学分析[J].中国水产科学,2023,30(8):1042-1053,12.基金项目
浙江省农业重大技术协同推广项目(2020XTTGSC03,2022XTTGSC04) （2020XTTGSC03,2022XTTGSC04）
象山县科技计划项目(2022C1001). （2022C1001）
