鸭呼肠孤病毒人工感染SPF鸡胚的病理学研究

doi:10.3864/j.issn.0578-1752.2016.14.019

摘要/Abstract

摘要： 【目的】前人研究表明禽呼肠孤病毒可以经卵垂直传播, 禽呼肠孤病毒能引起雏禽的病毒性关节炎、呼吸道和肠道疾病、肝炎、心肌炎、免疫抑制等。以实验室分离的鸭呼肠孤病毒HP080421株为研究对象，该毒株引起病变主要以鸭软脚为主要临床特征，病鸭肝脏和脾脏表面有大量的白色坏死灶、肾脏肿大、出血为主要病变特点。研究了鸭呼肠孤病毒对鸡胚的致病性,通过雏鸡的病理变化特点,探讨所分离的HP080421鸭呼肠孤病毒株是否也可感染鸡群，旨在为鸡场鸭呼肠孤病毒感染的防控提供理论依据。【方法】运用华中农业大学兽医病理学实验室分离鉴定的HP080421株鸭呼肠孤病毒通过尿囊腔接种SPF鸡胚，建立SPF鸡胚鸭呼肠孤病毒感染模型，培养鸡胚至雏鸡出壳，运用临床观察、病理剖检、H.E染色和免疫组织化学染色等病理组织学检查方法，对鸭呼肠孤病毒感染SPF鸡胚进行病理学研究和致病性分析。【结果】研究发现病毒感染鸡胚孵化培养至22—23日龄均啄壳，但不能自主出壳，需人工剥壳。病理剖检可见雏鸡的肝脏、脾脏肿胀质脆，表面及实质分布有黄白色坏死灶；脑部组织肿胀，并可见出血点和出血斑。H.E染色的组织病理学观察可见接毒组雏鸡肝脏、脾脏的坏死灶周围有大量淋巴细胞浸润；法氏囊的滤泡髓质部和胸腺的髓质部淋巴细胞减少、缺失；脑、肺脏、肾脏存在不同程度的淤血、出血和水肿；免疫组织化学染色结果表明：病毒阳性信号主要分布于肝脏、脾脏、肺脏和法氏囊等器官，并且位于上皮细胞和巨噬细胞的细胞质和胞核内。【结论】鸭呼肠孤病毒株HP080421 能够在SPF鸡胚中复制增殖，并使雏鸡主要器官发生明显的病理变化，病理变化主要集中在肝脏和淋巴器官，因此鸭呼肠孤病毒可通过垂直传播感染鸡群，还可导致雏鸡免疫抑制。本研究通过SPF鸡胚鸭呼肠孤病毒感染模型，阐述了鸭呼肠孤病毒感染鸡群的可能性，因此在实际养殖过程中，养殖户要防止鸭呼肠孤病毒对鸡胚的污染，切断传播途径，以达到预防鸡群感染鸭呼肠孤病毒的目的。

关键词: 鸭呼肠孤病毒, SPF鸡胚, 病理剖检, 免疫器官, H.E染色

Abstract: 【Objective】Previous studies showed that avian reovirus could infect vertically through egg, and avian reovirus can cause avian viral arthritis, respiratory and intestinal disease, myocarditis, hepatitis, immune suppression and so on. HP080421 strain of duck reovirus(DRV) isolated in the authors’ laboratory can cause soft duck feet as the main clinical features, and sick duck showed that a lot of white necrotic stove was on the surface of liver and spleen, and also kidney swelling and bleeding as the main pathological features. In this study, the pathogenicity of DRV to chicken embryo was investigated and whether the isolated HP080421 strain could infect chickens through the pathological changes of chicken was discussed, in order to provide a theoretical basis for the prevention and control of DRV infection.【Method】The infection model of SPF chicken to DRV was established through allantoic cavity inoculation SPF chicken embryos by using the isolated and identified HP080421 strains of DRV isolated in the lab. After chicks hatched from embryos, pathological examination methods such as clinical observation, pathological section examination, HE staining and immunohistochemical staining were used to study the pathobiology and pathogenicity of the SPF chicken embryo infected by DRV.【Result】Clinical observation found that chicken embryos were able to peck the shell after 22 - 23 days, but could not go out from the eggshell by themselves compared to the control group. At necropsy, liver and spleen were breakable and slightly swelling, many different size and yellow-white necrotic foci were consistently observed in the spleen and liver of the experimental group; the brain tissue was slightly swelling with few bleeding spots covered on it. Histopathological examination of H.E staining revealed necrotic foci in spleen and liver, which consisted of a necrotic center with lymphocytes infiltration at the periphery; in the Bursa of Fabricus, as well as in the thymus, lymphocyte depletion was apparent and cavities had developed in medulla. Besides, other organs such as lung, brain and kidney, showed different degrees of congestion and edema. Immunohistochemical detection showed that liver, lung, spleen and bursa of fabriciusa had positive signals, and were located in the cytoplasm and nucleus of epithelial cells and macrophages.【Conclusion】The results showed that the virus strain HP08421 could infect SPF chicken embryo and cause some specific pathologic changes. The pathological changes mainly focus on the liver and lymphoid organs, so DRV can be infected by vertical transmission of chickens, and can also lead to immune suppression. This study has expounded the possibility of infection of chickens by DRV, through the infection model of SPF chicken to DRV, as a matter of fact, in the actual process of production, farmers should prevent chicken embryo pollution by DRV to cut off the route of transmission, to achieve the purpose of preventing DRV infection.

Key words: duck reovirus, SPF chicken embryo, necropsy, immune organs, hematoxylin-eosin staining

刘晓丽，刘婷，刘波，程国富，谷长勤，张万坡，胡薛英. 鸭呼肠孤病毒人工感染SPF鸡胚的病理学研究[J]. 中国农业科学, 2016, 49(14): 2844-2849.

LIU Xiao-li, LIU Ting, LIU Bo, CHENG Guo-fu, GU Chang-qin, ZHANG Wan-po, HU Xue-ying. The Pathogenicity of Duck Reovirus on SPF Chicken Embryo[J]. Scientia Agricultura Sinica, 2016, 49(14): 2844-2849.

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