南苏门答腊岛巨港市孔雀分离的新城疫病毒的鉴定

L. Angeliya, Akbar Agus Anshori Mussama, S. Alawiyah, T. Guntoro, E. A. Srihanto, Y. P. Kristianingrum, W. Asmara, M. H. Wibowo
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摘要

简介:新城疫病是由新城疫病毒(NDV)引起的一种禽类传染病。印度尼西亚已报告在商业和后院饲养的鸡、鸽子、鸭和鹅,甚至鹰和孔雀中发生新城疫病例。孔雀是一种野生鸟类,受印度尼西亚1999年第7号政府条例保护。本研究旨在对孔雀ND病例进行NDV的分子鉴定、分离和鉴定。方法:对出现神经症状、腹泻后死亡的孔雀进行肺、脾等脏器取样。采用实时荧光定量pcr技术鉴定孔雀死因。对鸡胚蛋进行了病毒分离和胚胎变化及死亡观察。对NDV的F和HN全基因进行了测序。利用MEGA-X软件对核苷酸序列进行分析,包括氨基酸预测、氨基酸水平遗传变异分析、同源性分析和系统发育树构建。结果:样品鉴定结果为新城疫病毒阳性。观察到鸡胚胎发育迟缓,羽毛稀少,出血,在不到60小时内死亡。获得病毒分离,滴度为26。分子分析显示,F基因的RRQKRF切割位点模式同源性为95.8 ~ 97.6%,与印尼先前发现的ND病毒处于同一分支区。HN基因的抗原位点、糖酵解和中和表位均未发现氨基酸突变。结论:从孔雀中分离到的病毒是NDV的一种速度源毒株,亚基因型为VII.2,与以前在商品家禽和家禽中报道的NDV具有密切的遗传范围。这一结果表明,ND对受保护的野生鸟类也是一种威胁。
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Characterization of Newcastle Disease Virus Isolated from Peacocks in Palembang City, South Sumatra
Introduction: Newcastle Disease (ND) is an infectious disease in various types of poultry caused by the Newcastle Disease Virus (NDV). Cases of ND in Indonesia have been reported in commercial and backyard chickens, pigeons, ducks and geese, even in eagles and peacocks. Peacock is a wild bird protected by Indonesia's Government Regulation No. 7 of 1999. This study aims to identify, isolate and characterize the NDV molecularly in cases diagnosed as ND in peacocks. Method: Samples were obtained from organs (lungs and spleen) of peacocks which showed neurological symptoms, diarrhoea and then died. Real-time RT-PCR ND was used to identify the cause of death of the peacock. Virus isolation and observation of embryonic changes and death were carried out on embryonic chicken eggs. Sequencing was carried out to characterize the F and HN entire genes of the NDV. The nucleotide sequences were analyzed using MEGA-X software, including amino acid prediction, analysis of genetic variation at the amino acid level, homology and construction of the phylogenic tree. Result: The results of the sample identification were positive for the Newcastle disease virus. Observations of chicken embryos are stunted, have few feathers, are haemorrhagic, and die in less than 60 hours. Virus isolation was obtained with a titer of 26. Molecular analysis showed that the RRQKRF cleavage site pattern in the F gene had homology of 95.8-97.6% and was in the same branching area as the previous ND virus in Indonesia. There were no amino acid mutations at the antigenic site, glycolysis and neutralization epitopes in the HN gene. Conclusion: The virus isolated from the peacock is a velogenic strain of NDV, subgenotype VII.2 and has a close genetic range to the NDV that has been previously reported in commercial and domestic poultry. This result shows that ND is also a threat to protected wild birds.
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