文章摘要
四川地区养殖鲫鱼鲤疱疹病毒II型的鉴定及病理学研究
Identification and pathological study of Cyprinid herpesvirusⅡin Sichuan
投稿时间:2019-07-11  修订日期:2019-10-22
DOI:
中文关键词: 鲫鱼  鲤疱疹病毒2型  组织病理  超微病理
英文关键词: Crucian carp (Carassius auratus)  Cyprinid herpesvirusⅡ(CyHV-2)  Histopathology  Ultrastructure pathology
基金项目:四川省留学回国人员科技活动项目择优资助;四川省教育厅基础研究项目(17ZB0345)
作者单位E-mail
周瑶佳 四川农业大学动物医学院 zhouyaojia163@163.com 
田思璐 四川农业大学动物医学院  
许佳雪 四川农业大学动物医学院  
李尹麒 四川农业大学动物医学院  
杨悦 四川农业大学动物医学院  
耿毅 四川农业大学动物医学院  
黄小丽 四川农业大学动物科技学院  
陈德芳 四川农业大学动物科技学院  
汪开毓 四川农业大学动物医学院  
欧阳萍 四川农业大学动物医学院 ouyang.ping@live.cn 
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中文摘要:
      2018年10月,四川省自贡市某鲫鱼养殖场暴发传染性疾病,死亡率高达80%。【目的】为探究疾病病因和流行特点,【方法】对病鱼进行剖检、病理学观察、分子生物学检测及人工感染试验。【结果】患病鲫鱼临床表现为离群独游、浮头、全身发黑、体表出血。剖检发现鳃、肝脏、肾脏等器官出血、肿胀和坏死, 鳔点状出血。组织病理观察显示鳃呼吸上皮细胞肿胀、坏死及脱落;肾脏局灶性坏死;脾脏组织广泛变性坏死,造血系统崩解;肝脏发生空泡变性和坏死。透射电镜观察到脾脏和肾脏中存在4种不同大小的病毒颗粒,分别为DNA病毒内核、空衣壳、实心衣壳和有包膜的成熟病毒粒子。未成熟的病毒粒子存在于细胞核,成熟的病毒粒子存在于细胞质。电镜下可见鲤疱疹病毒II型(CyHV-2)在细胞核内完成核酸复制和核衣壳装配,囊膜蛋白在出核后获得。细胞核萎缩、核染色质边缘化,线粒体肿胀、坏死、嵴断裂,细胞质空泡化。取病鱼组织匀浆滤菌后接种到EPC及FHM细胞系,盲传3代后未见细胞病变效应。通过腹腔注射病鱼组织匀浆进行人工感染试验,试验组鲫鱼表现出与自然感染鲫鱼相同的临床症状而对照组鲫鱼正常,试验组累积死亡率为80%。用PCR方法检测CyHV-2的解旋酶基因片段,扩增出预期大小相符的目的条带。利用邻近法对该病毒的解旋酶基因进行系统发育分析,结果显示与CyHV-2(YZ-01)的同源性为99%。【结论】以上试验结果证实了CyHV-2是这次疾病暴发的原因。本研究首次报道了四川地区养殖鲫鱼CyHV-2感染引起大量死亡,并进行了系统的病理学观察,为CyHV-2致病机制和流行病学调查的研究提供了重要依据。
英文摘要:
      In October 2018, a crucian carp (Carassius auratus) farm located in Zigong, Sichuan Province, developed a communicable disease with a mortality rate of 80%. 【Purpose】In order to explore the cause of this disease, 【Method】sick fish were subjected to necropsy, pathological observation, molecular biological examination and artificial infection test. 【Results】 The clinical manifestations of diseased fish include leave the shoal of fish to swim alone; stay on the water surface; bodies blacken and body surface have haemorrhages. The necropsy showed haemorrhage, swelling and necrosis of organs such as gills, liver and kidney; swimming bladder have a lot of blutene chloaide. Histopathological changes show that the respiratory epithelial cells of gill were swollen, necrosis and sloughting. The kidneys show focal necrosis. The spleen tissue was extensively degenerated and necrotic, and the hematopoietic system collapses. Vacuolar degeneration and necrosis occur in the liver. Transmission electron microscopy showed that there were four different sizes of virus particles in the spleen and kidney, which were virus DNA core, empty viral capsid, solid capsid and enveloped mature virions. It can be seen that immature virions are present in the nucleus, and mature virions are present in the cytoplasm. Cyprinid herpesvirus Ⅱ (CyHV-2) completes nucleic acid replication and nucleocapsid assembly in the nucleus, and the envelope protein is obtained after transmembrane membrane. After filtering the diseased fish tissue homogenate, it was inoculated into EPC and FHM cell lines, and there was no cytopathic effect after 3 generations of blind transmission. The helicase gene fragment of CyHV-2 was amplified by PCR, and a positive band appeared. The artificial infection test was carried out by intraperitoneal injection of tissue homogenate. The test group showed the same clinical symptoms as the natural infection fish and the control group was normal. The cumulative mortality rate of the test group was 80%. Phylogenetic analysis of helicase gene by the neighbor-joining method revealed that the homology with CyHV-2 (YZ-01) was 99%. 【Conclusion】The results demonstrated that CyHV-2 was contributed to this outbreak disease. This study reported an outbreak disease caused by CyHV-2 in cultured crucian carp in Sichuan and systematic pathological was studied, which provided an important basis for study the pathogenesis and epidemiology of CyHV-2.
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