2004年初广东省高致病性禽流感(H5N疫点野生鸟类禽流感研究简报

2004年初禽流感疫情期间,针对广东省疫点地区开展了野生鸟类禽流感病毒(H5、H7、H9)调查取样工作,共获得血样76份、拭子125份、器官样21份。血清检测采用琼脂扩散(AGP)和血凝抑制试验(HI),病毒检测用鸡胚法鉴定。分析结果:(1)只在非疫点的汕头南澳的八哥上检测到H9病毒;(2)雷州、罗定、海丰及汕头4个地区的血清检测结果中H5和H9阳性率分别为31·6%和23·7%;沿海地区阳性率极显著高于非沿海地区;(3)候鸟的阳性率极显著高于留鸟。因此,该次疫情中候鸟中存在传播禽流感病毒的可能性。     

2004年初广东省高致病性禽流感(H5N1)疫点野生鸟类禽流感研究简报

2004年初禽流感疫情期间，针对广东省疫点地区开展了野生鸟类禽流感病毒(H5、H7、H9)调查取样工作，共获得血样76份、拭子125份、器官样21份。血清检测采用琼脂扩散(AGP)和血凝抑制试验(HI)，病毒检测用鸡胚法鉴定。分析结果：(1)只在非疫点的汕头南澳的八哥上检测到H9病毒；(2)雷州、罗定、海丰及汕头4个地区的血清检测结果中H5和H9阳性率分别为31.6%和23.7%；沿海地区阳性率极显著高于非沿海地区；(3)候鸟的阳性率极显著高于留鸟。因此，该次疫情中候鸟中存在传播禽流感病毒的可能性。     

2004年下半年广东野生鸟类禽流感研究初报

2004年下半年我们对广东省的野生鸟类进行禽流感取样调查,共取得血样263份、拭子样337份,血清检测采用琼脂扩散（AGP）和血凝抑制试验（HI）,病毒检测用鸡胚法鉴定。结果：1）病毒检测中分别在放生的白眉鸭（Arias querquedula）和石鸡（Alectoris chukar）体内检测到H5和H9病毒各1例;2）血清检测中H5的总阳性率为97.7％,而H9达98．5％,其中,不论是候鸟与留鸟间,还是放生个体、市场个体及野外个体间在H5差异不大且都高于94％,H9也是如此。由此,我们认为：1）鸟类放生存在传播禽流感的可能性;2）该时期野生鸟类中普遍出现H5和H9的高阳性率现象,可能存在H5和H9在野外的大范围传播情况。     

2012-2015年广东省活禽市场外环境禽流感病毒污染状况研究

研究广东省活禽市场外环境禽流感病毒污染状况并及时发现人流感发病潜在的危险因素,为人流感防治提供科学参考依据。应用传染病技术监测平台信息管理系统数据,采用描述性流行病学方法分析各种亚型病毒感染的流行病学特征,研究2012-2015年广东省活禽市场外环境禽流感病毒污染。共采集检测广东省21个地市级样本33079份,FluA 总阳性率为24．23％,H5、H7和 H9型高致病性禽流感病毒阳性率分别为3．70％、3．89％和13．53％;除2012年阳性率呈现季节性增加外,其他年份 FluA 核酸检测阳性率均在冬春季出现一个高峰。不同部位或地点采集的标本中,宰杀或摆放禽肉案板表面阳性率最高（FluA39．49％,H58．41％,H77．41％,H923．84％）,而采集的粪便标本阳性率最低（FluA14．99％,H51．73％、H72．38％、和 H97．23％）;所采集的标本所对应的相关动物种类中,鸡（64．08％）、鸭（55．84％）和鸟类（51．92％）的禽流感病毒阳性率都达到50％以上,H5、H7和 H9在各禽类中均可以检出。同时发现,在环境中检出 H7亚型多的地区分布与其相应地区 H7N9感染的病例数呈显著相关性,（r ＝0．689,P ＜0．05）;对2322份样本进行 H6亚型核酸检测,总阳性率为2．58％,并选取 H5、H6和 H9亚型标本153份进行 N 亚型检测,检测出 H5N1、H5N2、H5N6、H6N2和 H9N2等多种亚型。2012-2015年广东省21个地市活禽市场均存在 HA 亚型（H5、H7、H9和 H6）和 NA 亚型（N1、N2、N6）等多种亚型的污染,污染程度呈现季节性分布,不同样本类型和禽类其禽流感病毒分状况不同,H7亚型的污染严重程度与 H7N9的病例感染数呈正相关性。     

2013-2014年湖北省外环境中禽流感病毒污染情况分析

为了解2013-2014年湖北省外环境中禽流感病毒污染情况,采用实时荧光定量聚合酶链反应(polymerase chain reaction,PCR)对全省7个监测点1 642份活禽相关外环境标本进行禽流感病毒型别与阳性率监测.结果显示,2013-2014年湖北省外环境中禽流感病毒平均阳性检出率为19.1％,检出的禽流感病毒型别以H9亚型为主(占57.3％),未检出H7亚型.不同地区和季节禽流感病毒阳性率有所差异,鄂西北冬春季病毒检出阳性率高于鄂西南和江汉平原;家禽屠宰加工厂检出阳性率最高;阳性率最高的标本来自清洗禽类的污水.结果提示,湖北地区外环境中禽流感病毒阳性率较高,且型别多样,家禽屠宰加工人员对禽流感病毒有较高的暴露风险,需加强健康宣教,提高个人防护意识.     

1997～2006年广州市人禽流感监测分析

目的 在禽畜饲养、销售、宰杀场所和发生人禽流感疫情的疫点开展人感染禽流感的监测,探索禽流感病毒感染人的流行病学特征。方法 采集广州市禽流感相关高危人群的血清样本,用微量血凝抑制半加敏法进行检测。对监测到的H9N2阳性病例和H5N1的感染者进行流行病学调查。结果 近年来广州市禽接触职业人群H9感染率为4．56％(0．51％～10．00％),各年龄组差异无显著性;H5N1患者有多个售卖活禽肉菜市场活动史,1例H9N2患者及1例H5N1感染者无直接接触禽类活动史,另1例H5N1感染者为活禽交易市场禽类宰杀人员。结论 H5N1和H9N2可在相关高危人群造成隐性感染或轻症感染;除禽饲养场所外,农贸市场的活禽销售、宰杀场所也存在禽流感病毒感染的风险。     

2009～2013年我国活禽市场环境样本中禽流感病毒的检测

为了解中国活禽市场环境样本中禽流感病毒分布情况,对全国2009~2013年采自活禽市场的环境样本进行流感病毒核酸检测,并利用无特殊致病原鸡胚(SPF鸡胚)对A型流感病毒核酸阳性标本进行病毒分离。结果显示,环境标本中禽流感病毒核酸阳性率的波动呈明显的季节性趋势,冬春季节核酸检测阳性率较高;我国南方地区活禽市场环境样本的禽流感病毒核酸阳性率高于北方。市场中清洗禽类的污水和宰杀或摆放禽肉案板表面的擦拭样本核酸阳性率高于其它类型的样本。2009~2013年环境样本中分离到的禽流感病毒以H5和H9亚型为主,2013年之前H5亚型多于H9亚型,而2013年H9亚型超过了H5亚型。本研究表明,对城乡活禽市场中禽流感病毒的实时监测具有重要的公共卫生意义,可为我国人感染禽流感病毒的防控和预测预警提供依据。     

一种更灵敏的特异性检测H9亚型禽流感抗体的间接ELISA方法的建立

H9亚型禽流感在全球范围内广泛流行,控制其传播需监测H9亚型禽流感病毒的感染情况及疫苗的免疫效果。为了建立便于检测且灵敏特异的H9亚型禽流感抗体间接ELISA方法,本实验利用不同亚型之间变异较大的H9亚型禽流感病毒HA蛋白的头部球状区作为包被抗原,确定了最佳复合封闭液和抗体稀释液,提高了其特异性。结果显示建立的ELISA方法灵敏度高于血凝抑制试验(HI),且与H3N2、H5N2、H7N9亚型流感病毒及新城疫病毒(NDV)、鸡传染性支气管炎病毒(IBV)、鸡传染性法氏囊病毒(IBDV)和产蛋下降综合征病毒(EDSV)的阳性血清均无交叉反应。另外,利用该方法及HI试验对200份临床鸡血清样本进行检测,两种检测方法的符合率达97%,且存在较高的相关性(R2=0.981 1)。     

H5N1禽流感病毒血凝素抗原快速检测试剂对不同现场标本的检测比较

利用H5亚型禽流感病毒血凝素抗原快速检测试剂“H5-HA(Ag)Dot-ELISA(H5-Dot)”对来自陆禽、水禽的484份气管拭子、泄殖腔拭子和粪便拭子标本进行检测,结果:①不同采集方式的H5N1阳性标本的检出率高低有别,气管拭子的检出率最高,泄殖腔拭子次之,粪便拭子最低(P<0·05),因此建议现场采样时应尽可能采集气管拭子标本;②陆禽标本检出率显著高于水禽标本(P<0·05),对病毒培养阳性的陆禽气管拭子检出率达80%(95%CI:70·6%~87·8%),对水禽气管拭子标本的检出率为38%(95%CI:26·9%~49·4%),可能与标本中病毒滴度高低有关;③有症状与无症状陆禽标本的检出率无显著差异。另外,H5-Dot试剂对333份非H5病毒气管拭子标本的特异性为99·4%(95%CI:97·9%~99·9%)。这些结果表明,H5-Dot是一种较为可靠的H5亚型禽流感病毒早期快速检测方法,在缺乏仪器设施和高素质专业技术人员的H5N1禽流感病毒防控第一线具有重要推广价值。     

苏州活禽市场禽流感病毒(H7N9)季节变化及HA、NA、PB2基因系统进化分析

自2013年3月中国首次发现新型禽流感病毒H7N9以来,其于2013-2014年期间发生流行,2015年也有散发性感染。该病毒的流行不仅危及家禽养殖业,还对公共卫生安全造成严重威胁。为调查活禽市场中H7N9的进化史和季节性变化,本研究于2013年7-12月在H7N9主要流行地区之一江苏省苏州市活禽市场采集2 655份鸡、鸭咽拭子样本,对样本中流感病毒核酸进行检测。结果显示,冬季样本中H7N9阳性率显著高于夏季样本,同时发现样本中存在H5、H7和H9亚型毒株之间的混合感染。进一步对H7N9阳性样本的HA、NA和PB2基因序列进行分析,结果表明阳性样本中HA、NA和PB2基因序列与新型H7N9病毒的相应基因序列同源,其在家禽体内传代时也在继续进化。特别是一些样品中PB2基因序列与H5N1病毒PB2基因序列的同源性较高。结果提示,苏州存在一种新型H7N9病毒基因重排的可能性,建议在活禽市场对所有禽流感病毒亚型进行持续监控,从而有助于流感病毒的及时防控。     

H5N1高致病性禽流感病毒的危害及生态问题

高致病性禽流感(HPAI)H5N1病毒亚型已对人类健康、养殖业发展、野生鸟类及生态环境带来极大危害,引起国内外广泛关注。研究发现,禽流感病毒通过发生重组或者突变,可产生感染人类或其他生物的新病毒亚型,或产生更高的致病性,而人类亦具有丰富的与H5N1结合的受体。对候鸟迁徙停歇地禽流感调查表明,湿地、湖泊可能是HPAI病毒存活、散播的疫源地,病毒可随着鸟类的迁徙到处传播。因此,野生鸟类及其赖以生存的主要湿地环境处于感染HPAI的风险之中。     

Influenza a viruses from wild birds in Guatemala belong to the North American lineage

The role wild bird species play in the transmission and ecology of avian influenza virus (AIV) is well established; however, there are significant gaps in our understanding of the worldwide distribution of these viruses, specifically about the prevalence and/or significance of AIV in Central and South America. As part of an assessment of the ecology of AIV in Guatemala, we conducted active surveillance in wild birds on the Pacific and Atlantic coasts. Cloacal and tracheal swab samples taken from resident and migratory wild birds were collected from February 2007 to January 2010.1913 samples were collected and virus was detected by real time RT-PCR (rRT-PCR) in 28 swab samples from ducks (Anas discors). Virus isolation was attempted for these positive samples, and 15 isolates were obtained from the migratory duck species Blue-winged teal. The subtypes identified included H7N9, H11N2, H3N8, H5N3, H8N4, and H5N4. Phylogenetic analysis of the viral sequences revealed that AIV isolates are highly similar to viruses from the North American lineage suggesting that bird migration dictates the ecology of these viruses in the Guatemalan bird population.     

Molecular Characterization of Subtype H11N9 Avian Influenza Virus Isolated from Shorebirds in Brazil

Migratory aquatic birds play an important role in the maintenance and spread of avian influenza viruses (AIV). Many species of aquatic migratory birds tend to use similar migration routes, also known as flyways, which serve as important circuits for the dissemination of AIV. In recent years there has been extensive surveillance of the virus in aquatic birds in the Northern Hemisphere; however in contrast only a few studies have been attempted to detect AIV in wild birds in South America. There are major flyways connecting South America to Central and North America, whereas avian migration routes between South America and the remaining continents are uncommon. As a result, it has been hypothesized that South American AIV strains would be most closely related to the strains from North America than to those from other regions in the world. We characterized the full genome of three AIV subtype H11N9 isolates obtained from ruddy turnstones (Arenaria interpres) on the Amazon coast of Brazil. For all gene segments, all three strains consistently clustered together within evolutionary lineages of AIV that had been previously described from aquatic birds in North America. In particular, the H11N9 isolates were remarkably closely related to AIV strains from shorebirds sampled at the Delaware Bay region, on the Northeastern coast of the USA, more than 5000 km away from where the isolates were retrieved. Additionally, there was also evidence of genetic similarity to AIV strains from ducks and teals from interior USA and Canada. These findings corroborate that migratory flyways of aquatic birds play an important role in determining the genetic structure of AIV in the Western hemisphere, with a strong epidemiological connectivity between North and South America.     

Investigation of Avian Influenza Infections in Wild Birds,Poultry and Humans in Eastern Dongting Lake,China

We investigated avian influenza infections in wild birds, poultry, and humans at Eastern Dongting Lake, China. We analyzed 6,621 environmental samples, including fresh fecal and water samples, from wild birds and domestic ducks that were collected from the Eastern Dongting Lake area from November 2011 to April 2012. We also conducted two cross-sectional serological studies in November 2011 and April 2012, with 1,050 serum samples collected from people exposed to wild birds and/or domestic ducks. Environmental samples were tested for the presence of avian influenza virus (AIV) using quantitative PCR assays and virus isolation techniques. Hemagglutination inhibition assays were used to detect antibodies against AIV H5N1, and microneutralization assays were used to confirm these results. Among the environmental samples from wild birds and domestic ducks, AIV prevalence was 5.19 and 5.32%, respectively. We isolated 39 and 5 AIVs from the fecal samples of wild birds and domestic ducks, respectively. Our analysis indicated 12 subtypes of AIV were present, suggesting that wild birds in the Eastern Dongting Lake area carried a diverse array of AIVs with low pathogenicity. We were unable to detect any antibodies against AIV H5N1 in humans, suggesting that human infection with H5N1 was rare in this region.     

Complete Genome Sequence of an H9N2 Avian Influenza Virus Isolated from Egret in Lake Dongting Wetland

We isolated a recombinant H9N2 avian influenza virus (AIV) from fresh egret feces in the Ardeidae protection region of the Dongting Lake wetland area in China, and it was designated A/Egret/Hunan/1/2012(H9N2). This is the first report of isolating H9N2 AIV from wild birds in the Dongting Lake wetland. Its eight gene segments are generated by reassortment of gene segments of different AIV subtypes. These results are helpful for understanding the epidemiology and evolution of AIV in wild birds during migration.     

三江自然保护区野生迁徙水禽携带禽流感病毒和新城疫病毒状况的监测

[目的]为了对途经三江保护区的野生迁徙水禽携带禽流感病毒(AIV)和新城疫病毒(NDV)的状况进行有效监测.[方法]在2005年10月、2006年4月、2006年10月3个候鸟的迁徙季节从三江保护区采集了158只野鸟的咽拭子和肛拭子样本.应用SPF鸡胚盲传、血凝和血凝抑制试验和RT-PCR等方法进行了病毒的分离和鉴定.[结果]结果共分离到20株AIV和13株NDV.20株AIV均来自2006年10月采集的样品,经常规血清学分型鉴定分为12个亚型,11个亚型来源于绿头鸭,分别为H2N2(2/20),H2N6(2/20),H3N4(1/20),H3N6(2/20),H3N7(2/20),H3N8(2/20),H6N2(2/20),H11N2(1/20),H11N3(1/20),H11N5(2/20),H11N6(1/20),另外一株来源于白眉鸭,为H5N2(1/20).13株NDV则来自3个迁徙季节的5种不同水禽采,其中包括绿头鸭(8/13),豆雁(1/13),白额雁(1/13),绿翅鸭(1/13)和鸳鸯(2/13).[结论]这一结果表明,拥有极大种群数量、在世界范围内广泛分布的绿头鸭,被认为可能是AIV和NDV最重要的自然宿主之一,并在病毒的传播上比其他野生鸟类具有更为重要的生态学意义.     

Ecological Factors Driving Avian Influenza Virus Dynamics in Spanish Wetland Ecosystems

Studies exploring the ecological interactions between avian influenza viruses (AIV), natural hosts and the environment are scarce. Most work has focused on viral survival and transmission under laboratory conditions and through mathematical modelling. However, more integrated studies performed under field conditions are required to validate these results. In this study, we combined information on bird community, environmental factors and viral epidemiology to assess the contribution of biotic and abiotic factors in the occurrence of low pathogenic AIV in Spanish wetlands. For that purpose, seven locations in five different wetlands were studied during two years (2007–2009), including seven sampling visits by location. In each survey, fresh faeces (n = 4578) of wild birds and water samples were collected for viral detection. Also, the vegetation structure, water physical properties of wetlands, climatic conditions and wild bird community composition were determined. An overall AIV prevalence of 1.7%±0.4 was detected in faecal samples with important fluctuations among seasons and locations. Twenty-six AIV were isolated from the 78 RRT-PCR positive samples and eight different haemagglutinines and five neuraminidases were identified, being the combination H3N8 the most frequent. Variation partitioning procedures identified the combination of space and time variables as the most important pure factor – independently to other factors – explaining the variation in AIV prevalence (36.8%), followed by meteorological factor (21.5%) and wild bird community composition/vegetation structure (21.1%). These results contribute to the understanding of AIV ecological drivers in Spanish ecosystems and provide useful guidelines for AIV risk assessment identifying potential hotspots of AIV activity.