水产学报2018，Vol.42Issue(4)：575-585,11.DOI:10.11964/jfc.20170410788
锦鲤疱疹病毒天津株的分离与鉴定
Isolation and characterization of a strain of koi herpesvirus from diseased koi in Tianjin
摘要
Abstract
To investigate the cause of sudden death of the koi(Cyprinus carpio),the moribund koi that showed typical symptoms including sunken eyes and head, rotted gill and haemorrhages on the skin were examined. No bacteria were isolated from spleen, liver and kindey of diseased fish by bacteriological examination, but a large number of viral nucleocapsid were found in gill tissue by electron microscopy observation. After filtration treat-ment, the gills, brain, liver, spleen, and kidney tissue suspension was inoculated to carp fin cells, Cytopathic ef-fects (CPE) were presented 7 days post co-cultivation. In artificial infection test, the death rate of fish by infecting tissue homogenate and culture liquid of infected cell reached 93.3% and 86.7% respectively. Histopathological analysis showed that the target organs included the gill, liver and kidney. Positive results were obtained from dis-eased koi according to a PCR diagnosis of koi herpesvirus (KHV) proposed by Office International des Epizooties (OIE), and high concentration of KHV was detected in gill and kidney tissue by fluorescence quantitative PCR. Based on neighbor-joining analyses of the TK gene sequences, phylogenetic tree was constructed and the result in-dicated that the KHV isolated in our study was Asian genotype isolate. Furthermore, according to the sequence alignment of Marker I and Marker II regions of KHV, the KHV detected in this study belongs to I++II–genotype, and was named KHV-TJ1601. The present study demonstrated that the I++II–genotype KHVwas isolated from the naturally infected fish in North China for the first time, and provided important material for virus genesis, evolu-tion, classification, and vaccine preparation.关键词
鲤/锦鲤疱疹病毒/分离鉴定/基因型/天津Key words
Cyprinus carpio/KHV/isolation and identification/genotype/Tianjin分类
农业科技引用本文复制引用
罗璋,郝爽,张振国,孟一耕,史谢尧,冯守明..锦鲤疱疹病毒天津株的分离与鉴定[J].水产学报,2018,42(4):575-585,11.基金项目
天津市自然科学基金(16JCYBJC30000) （16JCYBJC30000）
天津市科技计划项目(15ZXBFNC00290) （15ZXBFNC00290）
天津市农业科技成果转化与推广项目(201501030) （201501030）
天津市水产产业技术体系创新团队项目(ITTFR2017008)Natural Science Foundation of Tianjin (16 JCYBJC30000) （ITTFR2017008）
Science and Technology Program of Tianjin (15 ZXBFNC00290) （15 ZXBFNC00290）
Project for Achievement Transformation and Promotion of Agricultural Science and Technology of Tianjin (201501030) （201501030）
Project for Tianjin Fisheries Innovative Team (ITTFR2017008) （ITTFR2017008）
