| 胡鑫宇,孙敏华,曾庆航,等.禽腺病毒血清2型KM株发病模型的建立[J].广东农业科学,2023,(5-6):- |
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禽腺病毒血清2型KM株发病模型的建立 |
| Establishment of a Disease Model for chickens to Fowl Adenovirus 2 |
| 投稿时间：2023-03-29 修订日期：2023-05-06 |
| DOI： |
| 中文关键词: 禽腺病毒 动物模型 分离鉴定 FAdV-2 致病性 HHS |
| 英文关键词: avian adenovirus pathogenesis model Separation identification Avian adenovirus serotype 2 Pathogenicity HHS |
| 基金项目:国家重点研发计划（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型的发病模型，为FAdV-2免疫原性评价和疫苗效果评价建立标准。【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基因序列和Hexon基因序列进行遗传演化分析，使用DNAstar和Mega11软件分析分离株和其他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禽腺病毒血清2型后，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】成功从发病鸡组织样品中分离得到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核苷酸序列同源性相似度为98.9%和99.3%，推导出氨基酸同源性相似度分别为98.9%和98.8%。结果显示静脉注射感染途径的致病性高于肌肉注射，试验鸡出现了明显的肝炎-心包积液综合征（HHS），成功建立KM株发病模型：感染途径为静脉注射，感染剂量为108.0TCID50，感染后鸡群出现死亡，死亡率为50%，病死鸡剖检可见明显的病理变化，鸡群感染后7d泄殖腔排毒阳性。建立的KM株发病模型可以增加我们对FAdV-2致病性的理解，对抗病毒药物的检测和疫苗评价提供一定的参考价值，可以预防和减少FAdV-2的传播。【结论】首次验证了FAdV-2也能引发肝炎-心包积液综合征，建立了禽腺病毒血清2型感染SPF鸡的发病模型：通过静脉注射108TCID50剂量的KM株，6周龄SPF鸡出现死亡、黄绿色稀粪等明显临床症状，剖检可见明显的肝炎-心包积液，7d内泄殖腔拭子核酸100%阳性。 |
| 英文摘要: |
| 【 Objective 】 To establish a disease model of avian adenovirus serotype 2 and establish standards for the immunogenicity evaluation and vaccine efficacy evaluation of FAdV-2. 【 Method 】 Samples suspected of avian adenovirus infection were isolated and identified. Genetic evolution analysis was performed on the fiber and hexon gene sequences of the isolated strains. DNAstar and Mega11 software were used to analyze the isolated strains and other representative strains of FAdV, and nucleotide and amino acid homology comparisons were conducted. After determining that the isolated virus is avian adenovirus serotype 2, the pathogenicity of the isolated strain to 6-week-old SPF chickens under different infection pathways and doses was analyzed through animal experiments, and the optimal infection mode and dose for establishing a FAdV-2 serotype disease model were screened. 【 Result 】 One FAdV-2 serotype strain was successfully isolated from diseased chicken tissue samples, named KM strain. Genetic evolution analysis showed that KM strain and GX01 strain both belong to the FAdV-2 serotype in avian adenovirus group I, species D. The homology analysis results showed that the similarity between the Fiber gene and the Hexagon nucleotide sequence of KM and GX01 strains was 98.9% and 99.3%, respectively, and the deduced amino acid homology similarity was 98.9% and 98.8%, respectively. The results showed that the pathogenicity of intravenous injection infection was higher than that of intramuscular injection. The experimental chickens developed obvious hepatitis pericardial effusion syndrome (HHS), and a KM strain disease model was successfully established. The infection route was intravenous injection, with an infection dose of 108.0TCID50. After infection, the chickens died, with a mortality rate of 50%. The necropsy of the diseased chickens showed obvious pathological changes, and the chickens were positive for cloacal detoxification 7 days after infection. The established KM strain disease model can increase our understanding of the pathogenicity of FAdV-2, provide certain reference value for the detection of antiviral drugs and vaccine evaluation, and prevent and reduce the spread of FAdV-2. [Conclusion] It is the first time to verify that FAdV-2 can also cause hepatitis pericardial effusion syndrome, and establish a disease model of avian adenovirus serotype 2 infecting SPF chickens: by intravenous injection of KM strain with 108TCID50 dose, 6-week-old SPF chickens appear obvious clinical symptoms such as death, chartreuse loose stool, and autopsy shows obvious hepatitis pericardial effusion, and cloacal swab nucleic acid is 100% positive within 7 days. |
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