禽腺病毒血清2型分离鉴定及感染SPF鸡发病模型建立  

Isolation and Identification of Fowl Adenovirus Serotype 2 and Establishment of A Disease Model for SPF Chickens

作　　者：胡鑫宇 孙敏华 曾庆航 谢梓民 袁朝霞 廖明 HU Xinyu;SUN Minhua;ZENG Qinghang;XIE Zimin;YUAN Zhaoxia;LIAO Ming(College of Animal Science&Technology,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China;Institute of Animal Health,Guangdong Academy of Agricultural Sciences/Guangdong Key Laboratory of Livestock Disease Prevention/Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases,Ministry of Agriculture and Rural Affairs,Guangzhou 510640,China)

机构地区：[1]仲恺农业工程学院动物科技学院,广东广州510225 [2]广东省农业科学院动物卫生研究所/广东省畜禽疫病防治研究重点实验室/农业农村部禽流感等家禽重大疾病防控重点实验室,广东广州510640

出　　处：《广东农业科学》2023年第7期146-155,F0002,共11页Guangdong Agricultural Sciences

基　　金：国家重点研发计划项目(2022YFD1801000);“十四五”广东省农业科技创新十大主攻方向“揭榜挂帅”项目(2022SDZG02);广东省农业科学院创新基金-产业专项(202144);云南省廖明专家工作站(202105AF150077)。

摘　　要：【目的】建立禽腺病毒血清2型(FAdV-2)的发病模型,为评价FAdV-2的免疫原性和疫苗有效性提供参考。【方法】对疑似禽腺病毒感染的样品进行分离鉴定,对分离株Fiber基因和Hexon基因序列进行遗传演化分析,使用DNAStar和MEGA7.0软件对分离株和其他FAdV代表株进行核苷酸和氨基酸同源性比较。在确定分离到的病毒为FAdV-2后,通过动物实验分析分离株在不同感染途径和不同感染剂量下对6周龄SPF鸡的致病性,筛选出建立FAdV-2血清型发病模型的最佳感染方式和最佳感染剂量。【结果】从发病鸡组织样品中成功分离得到1株FAdV-2血清型毒株,命名为KM株。遗传演化分析表明,KM株和GX01株同属于禽腺病毒Ⅰ群D种中的FAdV-2血清型;同源性分析结果显示,KM株和GX01株的Fiber基因和Hexon基因核苷酸序列同源性分别为98.9%和99.3%,推导出氨基酸同源性分别为98.9%和98.8%。KM株静脉注射感染途径的致病性高于肌肉注射,试验鸡出现明显的心包积液-肝炎综合征;KM株发病模型的感染途径为静脉注射,感染剂量为108.0 TCID50,感染后鸡群死亡率为50%,剖检病死鸡可见明显的病理变化,鸡群感染后1 d泄殖腔排毒阳性。【结论】首次验证了FAdV-2能引发心包积液-肝炎综合征,建立了FAdV-2感染SPF鸡的发病模型,可为药物研发和疫苗评价提供一定参考,将有效预防和减少FAdV-2传播。【Objective】A disease model of fowl adenovirus serotype 2(FAdV-2)was established to provide a refference for evaluating the immunogenicity and vaccine effectiveness of FAdV-2.【Method】The samples suspected to be infected with avian adenovirus were isolated and identified.The genetic evolution of the Fiber and Hexon gene sequences of the isolates was analyzed.The nucleotide and amino acid sequences of the isolated strain and other FAdV representative strains were analyzed using DNAStar and MEGA7.0.After determining that the isolated virus was FAdV-2,the pathogenicity of the isolated strain was analyzed in 6-week-old SPF chickens under different infection routes and different infection doses in animal experiment.The best infection model and infection dose were selected to establish the FAdV-2 pathogenesis model.【Result】One FAdV-2 strain was successfully isolated from the infected chicken tissue samples,named KM strain.Genetic evolution analysis shows that KM strain and GX01 strain belong to the FAdV-2 of avian adenovirus species D of groupⅠ.The homology analysis shows that the nucleotide similarity of Fiber gene and Hexon gene between KM strain and GX01 strain is 98.9%and 99.3%,and the amino acid similarity is 98.9%and 98.8%,respectively.The pathogenicity of intravenous injection was higher than that of intramuscular injection,and the chickens showed obvious hepatitis hydropericardium syndrome(HHS).The infection route of the KM strain model was intravenous injection,and the infection dose was 108.0 TCID50.The chickens died after infection,with a mortality rate of 50%.Obvious pathological changes were found in the dead chickens at necropsy,and the chickens were positive for cloaca virus shedding at 1 d post infection.The established KM strain disease model provides a certain reference for vaccine development and vaccine evaluation,which will effectively prevent and reduce the transmission of FAdV-2.【Conclusion】It is verified that FAdV-2 can cause hepatitis hydropericardium syndrome,and a pathogenesis

关 键 词：禽腺病毒 动物模型 分离鉴定 禽腺病毒血清2型(FAd V-2) 致病性 心包积液-肝炎综合征 

分 类 号：S858.315.3[农业科学—临床兽医学]