2015-2016年鄱阳湖地区禽流感病毒分子流行病学研究
发布时间：2019-05-07 23:46
【摘要】：研究目的:调查2015-2016年冬春季鄱阳湖地区活禽市场、家禽散养户/养殖场和候鸟栖息地环境中禽流感病毒的流行情况,分析检测到的禽流感病毒基因多样性和来源,揭示禽流感病毒在活禽市场、家禽散养户/养殖场和候鸟栖息地之间的流动,评估该地区新型禽流感病毒出现的风险,为禽流感病毒生态学研究提供基础研究数据。研究方法:在鄱阳湖地区活禽市场、家禽散养户/养殖场和候鸟栖息地环境中收集标本,使用SPF鸡胚进行病毒分离,使用Real-Time RT-PCR鉴定病毒,进而利用深度测序技术获得病毒全基因组序列,随后重点对主要HA亚型、NA亚型和NS基因分别进行序列突变位点分析和系统进化分析,进一步分析病毒的起源和进化。主要结果:(1)2015-2016年冬春季,在鄱阳湖地区监测中发现了 23种亚型的禽流感病毒。其中在活禽市场和家禽散养户/养殖场中分离到了 1100株禽流感病毒,检测到22种亚型的禽流感病毒,其中H1N3、H2N3和H4N3亚型禽流感病毒是在鄱阳湖地区首次发现,H12N2亚型禽流感病毒是在中国首次发现。在家禽养殖场环境标本中发现了 H7N9亚型病毒。在候鸟栖息地环境标本中检出了 7株病毒,包括H1N1、H3N2/N6、H5N1和H5N6亚型病毒。(2)候鸟-家禽养殖场-活禽市场之间,尤其是家禽养殖场-活禽市场环境之间存在基因流动现象,并发现一些禽流感病毒如H1N3、H2N3、H6N2、H6N5、H10N3和H12N2亚型禽流感首次进入到该地区的家禽环境中。(3)在该年度家禽中流行的禽流感病毒中,含有A等位基因和B等位基因NS片段的禽流感病毒同时流行,B等位基因所占比例低。(4)在H5N1、H5N2、H5N6、H7N9和H9N2病毒的M2蛋白中发现了金刚烷胺耐药性突变,在H3N6、H4N6、H5N6、H6N6、H7N3和H7N9病毒的NA蛋白上发现了NA功能性位点的突变,其生物学表型有待进一步验证。结论:2015-2016年度,鄱阳湖地区家禽养殖场和活禽市场环境被禽流感病毒污染的情况较为严重,高于既往监测年度。在鄱阳湖地区的活禽市场,家禽散养户/养殖场以及候鸟栖息地环境中的禽流感病毒监测中发现了 23种亚型的禽流感病毒,分离的多株低致病性禽流感病毒与东亚-澳大利亚迁徙路线上候鸟或者家禽中流行的禽流感病毒较为接近。而且禽流感病毒在活禽市场与家禽散养户/养殖场之间、在家禽环境与候鸟环境之间存在流动。在该地区的家禽养殖场环境中发现了 H5N6亚型和H7N9亚型禽流感病毒,因此需要在该地区持续开展禽流感病毒监测,为人感染禽流感病毒提供风险预警,并促进家禽生产中的生物安全措施的升级提供支持。同时,在多种亚型的禽流感病毒中发现了禽流感病毒的跨区域与和跨国传播,因此需要在禽流感病毒的监测和预防方面加强国际合作。
[Abstract]:Objective: to investigate the prevalence of avian influenza virus (AIV) in the winter and spring of 2015-2016 in Poyang Lake region, and analyze the genetic diversity and origin of avian influenza virus. To reveal the mobility of avian influenza virus (AIV) between live poultry market, poultry farm / farm and migratory habitat, to evaluate the risk of the emergence of new avian influenza virus in this area, and to provide basic research data for the ecological research of avian influenza virus (AIV). Methods: samples were collected in the living poultry market in Poyang Lake area, and the virus was isolated by SPF chicken embryo, and identified by Real-Time RT-PCR, and the samples were collected in the environment of fowls / fowls and migratory bird habitats in Poyang Lake area. Then the whole genome sequence of the virus was obtained by deep sequencing. Then the main HA subtype, NA subtype and NS gene were analyzed by sequence mutation site analysis and phylogenetic analysis, respectively, and the origin and evolution of the virus were further analyzed. Main results: (1) in the winter and spring of 2015-2016, 23 subtypes of avian influenza virus were found in Poyang Lake area. Among them, 1100 strains of avian influenza viruses were isolated from live poultry markets and domestic poultry farmers / farms, and 22 subtypes of avian influenza viruses were detected. Among them, H1N1, H2N3 and H4N3 subtype avian influenza viruses were first found in Poyang Lake region. H12N2 subtype avian influenza virus was first found in China. H7N9 subtype virus was found in environmental samples of poultry farms. Seven strains of viruses were detected in environmental specimens of migratory bird habitats, including H1, H3N2, H5N1 and H5N6 subtype viruses. (2) between migratory birds, poultry farms and live poultry markets. In particular, there is a phenomenon of gene flow between poultry farms and the living poultry market environment, and some avian influenza viruses such as H1N1, H2N3, H6N2, H6N5are found. H10N3 and H12N2 subtype avian influenza for the first time entered the poultry environment in the region. (3) among the avian influenza viruses prevalent in poultry during the year, avian influenza viruses containing NS fragments of the A allele and the B allele were prevalent at the same time. (4) mutations in the M2 proteins of H5N1, H5N2, H5N6, H7N9 and H9N2 were found to be resistant to amantadine, and mutations in the functional site of NA were found in H3N6, H4N6, H5N6, H6N6, H7N3 and H7N9 viruses NA proteins, (4) mutations in the M2 proteins of H5N1, H5N2, H5N6, H7N9, and NA were found in H3N6, H4N6, H5N6, H6N6, H7N3 and H7N9 viruses. Its biological phenotype needs to be further verified. Conclusion: in 2015 / 2016, the environment of poultry farms and live poultry markets in Poyang Lake area was seriously polluted by avian influenza virus, which was higher than that in previous surveillance years. Twenty-three subtypes of avian influenza viruses were found in the live poultry market in Poyang Lake region, in domestic poultry farms / farms and in the habitat environment of migratory birds. Several strains of low pathogenic avian influenza virus isolated are similar to those prevalent in migratory birds or poultry on East Asia-Australia migration routes. Moreover, avian influenza virus flows between live poultry market and domestic poultry farm, between poultry environment and migratory bird environment. Avian influenza viruses (H5N6 and H7N9) have been found in poultry farms in the region. Therefore, there is a need for continued surveillance of avian influenza viruses in the region to provide a risk warning for human infection with avian influenza viruses. And promote the upgrading of biosafety measures in poultry production to provide support. At the same time, the transregional and transnational transmission of avian influenza virus has been found in many subtypes of avian influenza virus, so it is necessary to strengthen international cooperation in the surveillance and prevention of avian influenza virus.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R511.7
本文编号：2471457
[Abstract]:Objective: to investigate the prevalence of avian influenza virus (AIV) in the winter and spring of 2015-2016 in Poyang Lake region, and analyze the genetic diversity and origin of avian influenza virus. To reveal the mobility of avian influenza virus (AIV) between live poultry market, poultry farm / farm and migratory habitat, to evaluate the risk of the emergence of new avian influenza virus in this area, and to provide basic research data for the ecological research of avian influenza virus (AIV). Methods: samples were collected in the living poultry market in Poyang Lake area, and the virus was isolated by SPF chicken embryo, and identified by Real-Time RT-PCR, and the samples were collected in the environment of fowls / fowls and migratory bird habitats in Poyang Lake area. Then the whole genome sequence of the virus was obtained by deep sequencing. Then the main HA subtype, NA subtype and NS gene were analyzed by sequence mutation site analysis and phylogenetic analysis, respectively, and the origin and evolution of the virus were further analyzed. Main results: (1) in the winter and spring of 2015-2016, 23 subtypes of avian influenza virus were found in Poyang Lake area. Among them, 1100 strains of avian influenza viruses were isolated from live poultry markets and domestic poultry farmers / farms, and 22 subtypes of avian influenza viruses were detected. Among them, H1N1, H2N3 and H4N3 subtype avian influenza viruses were first found in Poyang Lake region. H12N2 subtype avian influenza virus was first found in China. H7N9 subtype virus was found in environmental samples of poultry farms. Seven strains of viruses were detected in environmental specimens of migratory bird habitats, including H1, H3N2, H5N1 and H5N6 subtype viruses. (2) between migratory birds, poultry farms and live poultry markets. In particular, there is a phenomenon of gene flow between poultry farms and the living poultry market environment, and some avian influenza viruses such as H1N1, H2N3, H6N2, H6N5are found. H10N3 and H12N2 subtype avian influenza for the first time entered the poultry environment in the region. (3) among the avian influenza viruses prevalent in poultry during the year, avian influenza viruses containing NS fragments of the A allele and the B allele were prevalent at the same time. (4) mutations in the M2 proteins of H5N1, H5N2, H5N6, H7N9 and H9N2 were found to be resistant to amantadine, and mutations in the functional site of NA were found in H3N6, H4N6, H5N6, H6N6, H7N3 and H7N9 viruses NA proteins, (4) mutations in the M2 proteins of H5N1, H5N2, H5N6, H7N9, and NA were found in H3N6, H4N6, H5N6, H6N6, H7N3 and H7N9 viruses. Its biological phenotype needs to be further verified. Conclusion: in 2015 / 2016, the environment of poultry farms and live poultry markets in Poyang Lake area was seriously polluted by avian influenza virus, which was higher than that in previous surveillance years. Twenty-three subtypes of avian influenza viruses were found in the live poultry market in Poyang Lake region, in domestic poultry farms / farms and in the habitat environment of migratory birds. Several strains of low pathogenic avian influenza virus isolated are similar to those prevalent in migratory birds or poultry on East Asia-Australia migration routes. Moreover, avian influenza virus flows between live poultry market and domestic poultry farm, between poultry environment and migratory bird environment. Avian influenza viruses (H5N6 and H7N9) have been found in poultry farms in the region. Therefore, there is a need for continued surveillance of avian influenza viruses in the region to provide a risk warning for human infection with avian influenza viruses. And promote the upgrading of biosafety measures in poultry production to provide support. At the same time, the transregional and transnational transmission of avian influenza virus has been found in many subtypes of avian influenza virus, so it is necessary to strengthen international cooperation in the surveillance and prevention of avian influenza virus.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R511.7
【参考文献】
相关期刊论文 前1条
1 LI Xiao Dan;ZOU Shu Mei;ZHANG Ye;BAI Tian;GAO Rong Bao;ZHANG Xin;WU Jie;SHU Yue Long;;A Novel Reassortant H2N3 Influenza Virus Isolated from China[J];Biomedical and Environmental Sciences;2014年04期
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