禽腺病毒4型云南株分离鉴定及Hexon基因序列分析

doi:10.16431/j.cnki.1671-7236.2022.10.026

摘要/Abstract

摘要： 【目的】分析云南省家禽及野鸟禽腺病毒4型(Fowl adenovirus-4,FAdV-4)流行情况、Hexon基因差异性及致病性,为研究FAdV-4流行及Hexon基因对病毒毒力的影响提供参考。【方法】2020年1月―2021年6月在云南省昆明市、曲靖市、楚雄州、大理州、红河州、玉溪市等家禽养殖密集区采集疑似感染FAdV-4家禽肝脏组织样品158份,候鸟迁徙地采集野鸟新鲜粪便310份,采用PCR技术检测FAdV-4核酸,对获得的代表性阳性样品进行病原分离鉴定,并对分离毒株进行致病性及Hexon基因序列分析。【结果】家禽肝脏组织样品中检出FAdV-4核酸阳性样品19份,阳性率为12.03%(19/158);黑颈鹤粪便样品中检出核酸阳性样品1份,阳性率为0.3%(1/310)。从FAdV-4核酸阳性样品中分离到7株FAdV-4,致病性研究结果显示,FAdV-4云南分离株均能致鸡胚发育不良,引起4周龄肉鸡发生心包积液-肝炎综合征,鸡胚半数致死量(ELD₅₀)为10^―6.17~10^―4.32/mL。7株分离株均聚类于FAdV-C亚群,Hexon蛋白与国内FAdV-4高致病性毒株具有相同的¹⁸⁸R、¹⁹³R、¹⁹⁵Q特征。分离株感染SPF鸡后,临床症状明显,具有高致病性且高致死率,肝细胞病变明显。【结论】研究初步掌握了云南FAdV-4的流行情况,首次在黑颈鹤粪便中检测到FAdV-4病原核酸,分离获得7株FAdV-4,部分毒株Hexon蛋白存在氨基酸突变。

关键词: 禽腺病毒4型(FAdV-4); Hexon基因; 致病性; 遗传进化

Abstract: 【Objective】 This study was aimed to analyze the prevalence,genetic difference of Hexon gene and pathogenicity of Fowl adenovirus-4 (FAdV-4) in Yunnan province,so as to provide reference for the epidemiology of FAdV-4 and the influence of Hexon gene on virus virulence.【Method】 158 samples of poultry liver tissue suspected of FAdV-4 infection and 310 fresh feces of wild birds were collected from January 2020 to June 2021 in Kunming,Qujing,Chuxiong,Dali,Honghe and Yuxi of Yunnan province,FAdV-4 nucleic acid was detected by PCR,representative positive samples were isolated and identified,and pathogenic and Hexon gene sequence of isolated strains were analyzed.【Result】 In 158 poultry tissue samples,19 samples were positive for FAdV-4 pathogenic nucleic acid,with a positive rate of 12.03%,and 1 sample (Black-necked cranes) was positive for 310 fresh feces of wild birds,with a positive rate of 0.3%.7 strains of FAdV-4 were isolated from positive samples.The results of pathogenicity study showed that all strains of FAdV-4 in Yunnan could cause chicken embryo dysplasia and pericardial effusion-hepatitis syndrome in 4-week broilers.The embryo median lethal dose (ELD₅₀) was 10^―6.17 to 10^―4.32/mL.The 7 strains were clustered in the branches of FAdV-C subgroup,and Hexon protein had the same characteristics of ¹⁸⁸R,¹⁹³R and ¹⁹⁵Q as domestic highly pathogenic strains.The strains infected with SPF chickens showed obvious clinical symptoms,highly pathogenic,high lethality and significant hepatocellular lesions.【Conclusion】 The prevalence of FAdV-4 in Yunnan was preliminarily understood,FAdV-4 pathogenic nucleic acid was detected in the feces of Black-necked cranes for the first time,7 strains of FAdV-4 were isolated,and some strains of Hexon protein had amino acid mutations.

Key words: Fowl adenovirus-4 (FAdV-4); Hexon gene; pathogenicity; genetic evolution

中图分类号:

S852.65⁺7

薛晓岩, 张振兴, 曾毅, 季佳, 代红炀, 李素华, 宋建领. 禽腺病毒4型云南株分离鉴定及Hexon基因序列分析[J]. 中国畜牧兽医, 2022, 49(10): 3932-3942.

XUE Xiaoyan, ZHANG Zhenxing, ZENG Yi, JI Jia, DAI Hongyang, LI Suhua, SONG Jianling. Isolation and Identification of Yunnan Fowl Adenovirus-4 and Sequence Analysis of Hexon Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3932-3942.

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