| 胡鑫宇 1，2，孙敏华 2，曾庆航 1，2，谢梓民 2，袁朝霞1，廖 明 2.禽腺病毒血清 2 型分离鉴定及感染 SPF 鸡发病模型建立[J].广东农业科学,2023,50(7):146-155 |
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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 |
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| DOI：10.16768/j.issn.1004-874X.2023.07.015 |
| 中文关键词: 禽腺病毒 动物模型 分离鉴定 禽腺病毒血清 2 型（FAdV-2） 致病性 心包积液 - 肝炎综合征 |
| 英文关键词: avian adenovirus animal model isolation and identification fowl adenovirus serotype 2 (FAdV-2) pathogenicity hydropericardium hepatitis syndrome |
| 基金项目: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） |
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| 中文摘要: |
| 【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。 |
| 英文摘要: |
| 【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 group Ⅰ. 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 model of FAdV-2 infection in SPF chickens is established for the first time. |
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