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引用本文:袁雪梅,潘晓艺,郝贵杰,刘莉,吕孙建,蔺凌云,沈锦玉.一例异育银鲫(Carassius auratus gibelio)暴发性出血病病原分析.海洋与湖沼,2019,50(4):913-920.
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一例异育银鲫(Carassius auratus gibelio)暴发性出血病病原分析
袁雪梅, 潘晓艺, 郝贵杰, 刘莉, 吕孙建, 蔺凌云, 沈锦玉
农业农村部淡水渔业健康养殖重点实验室 浙江省鱼类健康与营养重点实验室 浙江省淡水水产研究所 湖州 313001
摘要:
2016年6月,江苏某异育银鲫(Carassius auratus gibelio)养殖场暴发一种传染性急性出血病,造成养殖银鲫大量死亡。为分析此次疾病病因及流行规律,本研究从发病养殖场采集患出血病的异育银鲫,从细菌、病毒及寄生虫三个方面对病原进行了分析。采用病原菌分离、组织病理学观察、超薄切片电镜观察、病毒核酸分析、回感实验等对病原进行鉴定。结果显示从发病鲫鱼体内分离到病毒一株,未发现寄生虫及细菌感染。经测序及序列分析,该病毒为鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2,CyHV-2)病毒,组织病理学观察结果显示患病鱼的鳃和肾脏有明显病变,电镜下可观察到病鱼脾脏组织有带囊膜的球形病毒,囊膜直径约为170—200nm,病毒衣壳直径约为110—120nm,核心直径约为60nm,用组织匀浆感染鲫鱼囊胚细胞系(CGB)可稳定地观察到典型的细胞病变,用患病鱼组织匀浆液人工感染异育银鲫的死亡率高达100%,荧光定量PCR检测到该病毒可感染多器官,其中以脾脏中病毒含量最高,其次是脑,肝脏中最少。本研究可为CyHV-2的诊断防控及疫苗研制提供资料。
关键词:  异育银鲫  暴发性出血病  病毒分离鉴定  人工感染
DOI:10.11693/hyhz20190100021
分类号:S917.1
基金项目:浙江省自然科学基金资助项目,LY16C190001号;浙江省重点研发计划,2019C02058号;2019年浙江省省属科研院所扶持专项,2019YSZX001号。
ANALYSIS OF PATHOGEN IN AN EXPLOSIVE HEMORRHAGE DISEASE OF CARASSIUS AURATUS GIBELIO
YUAN Xue-Mei, PAN Xiao-Yi, HAO Gui-Jie, LIU Li, LÜ Sun-Jian, LIN Ling-Yun, SHEN Jin-Yu
Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China
Abstract:
In June 2016, local farmers suffered from substantial losses of Carassius auratus gibelio, or gibel carp in yield due to an outbreak of an acute hemorrhagic disease in Jiangsu Province, China. To understand the prevalence and etiological factor of this disease, ill fish were collected and the pathogens including bacteria, virus, and parasite were diagnosed simultaneously. A comprehensive analysis in histopathology, pathogen isolation, quantitative real-time PCR (qRT-PCR), and transmission electron microscope (TEM) observation was conducted to identify the accurate pathogen. A strain of cyprinid herpesvirus Ⅱ (CyHV-2) was successfully isolated from an ill gibel carp in which serious lesion was observed in gill and kidney; however, no bacterium nor parasite was identified. In the TEM images, a spherical virion was clearly observed in spleen in nucleus diameter of 60nm, envelope diameter of 170-200nm, and capsid diameter of 110-120nm. In addition, C. auratus embryo cell line (CGB) was infected by virus and obvious cytopathic effect was observed. Artificial infection demonstrated that the virus injection led to 100% mortality rate; and the highest virus concentration was found in spleen. This research offers useful information beneficial to the development of vaccine and diagnostic products.
Key words:  Carassius auratus gibelio  acute septicemia  virus isolation and identification  artificial infection
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