Highly pathogenic avian influenza H5N1, Thailand, 2004

In January 2004, highly pathogenic avian influenza (HPAI) virus of the H5N1 subtype was first confirmed in poultry and humans in Thailand. Control measures, e.g., culling poultry flocks, restricting poultry movement, and improving hygiene, were implemented. Poultry populations in 1,417 villages in 60 of 76 provinces were affected in 2004. A total of 83% of infected flocks confirmed by laboratories were backyard chickens (56%) or ducks (27%). Outbreaks were concentrated in the Central, the southern part of the Northern, and Eastern Regions of Thailand, which are wetlands, water reservoirs, and dense poultry areas. More than 62 million birds were either killed by HPAI viruses or culled. H5N1 virus from poultry caused 17 human cases and 12 deaths in Thailand; a number of domestic cats, captive tigers, and leopards also died of the H5N1 virus. In 2005, the epidemic is ongoing in Thailand.     

Fatal avian influenza A H5N1 in a dog

Avian influenza H5N1 virus is known to cross the species barrier and infect humans and felines. We report a fatal H5N1 infection in a dog following ingestion of an H5N1-infected duck during an outbreak in Thailand in 2004. With new reports of H5N1 virus continuing across Asia, Europe, and Africa, this finding highlights the need for monitoring of domestic animals during outbreaks.     

Novel Reassortant Influenza A(H5N8) Viruses among Inoculated Domestic and Wild Ducks,South Korea, 2014

An outbreak of highly pathogenic avian influenza, caused by a novel reassortant influenza A (H5N8) virus, occurred among poultry and wild birds in South Korea in 2014. The aim of this study was to evaluate the pathogenesis in and mode of transmission of this virus among domestic and wild ducks. Three of the viruses had similar pathogenicity among infected domestic ducks: the H5N8 viruses were moderately pathogenic (0%–20% mortality rate); in wild mallard ducks, the H5N8 and H5N1 viruses did not cause severe illness or death; viral replication and shedding were greater in H5N8-infected mallards than in H5N1-infected mallards. Identification of H5N8 viruses in birds exposed to infected domestic ducks and mallards indicated that the viruses could spread by contact. We propose active surveillance to support prevention of the spread of this virus among wild birds and poultry, especially domestic ducks.     

Evaluation of a positive marker of avian influenza vaccination in ducks for use in H5N1 surveillance

Control measures for H5N1 avian influenza involve increased biosecurity, monitoring, surveillance and vaccination. Subclinical infection in farmed ducks is important for virus persistence. In major duck rearing countries, homologous H5N1 vaccines are being used in ducks, so sero-surveillance using H5- or N1-specific antibody testing cannot identify infected flocks. An alternative is to include a positive marker for vaccination. Testing for an antibody response to the marker would confirm approved vaccine use. Concurrent testing for H5 antibody responses would determine levels adequate for protection or indicate recent infection, with an anamnestic H5 antibody response requiring further virological investigation. In this study, we have evaluated the use of a TT marker in ducks given avian influenza vaccination. Wild or domestic ducks were tested for antibodies against TT and all 463 ducks were negative. High levels of TT-specific antibodies, produced in twice-TT vaccinated Muscovy ducks, persisted out to 19 weeks. There was no interference by inclusion of TT in an inactivated H6N2 vaccine for H6- or TT-seroconversion. Thus TT is a highly suitable exogenous marker for avian influenza vaccination in ducks and allows sero-surveillance in countries using H5N1 vaccination.     

Seroprevalence of Antibodies to Avian Influenza Virus A (H5N1) among Residents of Villages with Human Cases, Thailand, 2005

In 2005, we assessed the seroprevalence of neutralizing antibodies to avian influenza virus A (H5N1) among 901 residents of 4 villages in Thailand where at least 1 confirmed human case of influenza (H5N1) had occurred during 2004. Although 68.1% of survey participants (median age 40 years) were exposed to backyard poultry and 25.7% were exposed to sick or dead chickens, all participants were seronegative for influenza virus (H5N1).     

Avian influenza virus (H5N1) replication in feathers of domestic waterfowl

We examined feathers of domestic ducks and geese inoculated with 2 different avian influenza virus (H5N1) genotypes. Together with virus isolation from the skin, the detection of viral antigens and ultrastructural observation of the virions in the feather epidermis raise the possibility of feathers as sources of infection.     

Free-grazing ducks and highly pathogenic avian influenza, Thailand

Thailand has recently had 3 epidemic waves of highly pathogenic avian influenza (HPAI); virus was again detected in July 2005. Risk factors need to be identified to better understand disease ecology and assist HPAI surveillance and detection. This study analyzed the spatial distribution of HPAI outbreaks in relation to poultry, land use, and other anthropogenic variables from the start of the second epidemic wave (July 2004-May 2005). Results demonstrate a strong association between H5N1 virus in Thailand and abundance of free-grazing ducks and, to a lesser extent, native chickens, cocks, wetlands, and humans. Wetlands used for double-crop rice production, where free-grazing duck feed year round in rice paddies, appear to be a critical factor in HPAI persistence and spread. This finding could be important for other duck-producing regions in eastern and southeastern Asian countries affected by HPAI.     

Outbreaks of avian influenza A (H5N1) in Asia and interim recommendations for evaluation and reporting of suspected cases--United States, 2004

During December 2003-February 2004, outbreaks of highly pathogenic avian influenza A (H5N1) among poultry were reported in Cambodia, China, Indonesia, Japan, Laos, South Korea, Thailand, and Vietnam. As of February 9, 2004, a total of 23 cases of laboratory-confirmed influenza A (H5N1) virus infections in humans, resulting in 18 deaths, had been reported in Thailand and Vietnam. In addition, approximately 100 suspected cases in humans are under investigation by national health authorities in Thailand and Vietnam. CDC, the World Health Organization (WHO), and national health authorities in Asian countries are working to assess and monitor the situation, provide epidemiologic and laboratory support, and assist with control efforts. This report summarizes information about the human infections and avian outbreaks in Asia and provides recommendations to guide influenza A (H5N1) surveillance, diagnosis, and testing in the United States.     

Pekin and Muscovy ducks respond differently to vaccination with a H5N1 highly pathogenic avian influenza (HPAI) commercial inactivated vaccine

Domestic ducks are key intermediates in the transmission of H5N1 highly pathogenic avian influenza (HPAI) viruses, and therefore are included in vaccination programs to control H5N1 HPAI. Although vaccination has proven effective in protecting ducks against disease, different species of domestic ducks appear to respond differently to vaccination, and shedding of the virus may still occur in clinically healthy vaccinated populations. In this study we compared the response to vaccination between two common domestic duck species, Pekin (Anas platyrhynchos domesticus) and Muscovy (Cairina moschata), which were vaccinated with a commercial inactivated vaccine using one of three different schedules in order to elicit protection to H5N1 HPAI before one month of age. Clear differences in responses to vaccination were observed; the Muscovy ducks developed lower viral antibody titers induced by the same vaccination as Pekin ducks and presented with higher morbidity and mortality after challenge with an H5N1 HPAI virus. When comparing the response to infection in non-vaccinated ducks, differences were also observed, with infected Muscovy ducks presenting a lower mean death time and more severe neurological signs than Pekin ducks. However Pekin ducks had significantly higher body temperatures and higher levels of nitric oxide in the blood at 2 days post challenge than Muscovy ducks, indicating possible differences in innate immune responses. Comparison of the expression of innate immune related genes in spleens of the non-vaccinated infected ducks showed differences including significantly higher levels of expression of RIG-I in Pekin ducks and of IL-6 in Muscovy ducks. Both duck species showed an up-regulation of IFNα and MHC-I expression, and a down-regulation of MHC-II. In conclusion, differences in response to infection and vaccination were observed between the two domestic duck species. This information should be taken into account when developing effective vaccination programs for controlling H5N1 HPAI in different species of ducks.     

Evolution of H5N1 avian influenza viruses in Asia

An outbreak of highly pathogenic avian influenza A (H5N1) has recently spread to poultry in 9 Asian countries. H5N1 infections have caused > or =52 human deaths in Vietnam, Thailand, and Cambodia from January 2004 to April 2005. Genomic analyses of H5N1 isolates from birds and humans showed 2 distinct clades with a nonoverlapping geographic distribution. All the viral genes were of avian influenza origin, which indicates absence of reassortment with human influenza viruses. All human H5N1 isolates tested belonged to a single clade and were resistant to the adamantane drugs but sensitive to neuraminidase inhibitors. Most H5N1 isolates from humans were antigenically homogeneous and distinct from avian viruses circulating before the end of 2003. Some 2005 isolates showed evidence of antigenic drift. An updated nonpathogenic H5N1 reference virus, lacking the polybasic cleavage site in the hemagglutinin gene, was produced by reverse genetics in anticipation of the possible need to vaccinate humans.     

我国H7N9禽流感病毒血凝素和神经氨酸酶基因的分子进化分析

  目的  通过对安徽省安庆市人感染H7N9禽流感外环境标本检测,分析外环境中病毒污染情况,研究判断人感染H7N9的风险,为制定防控措施提供依据。  方法  于2017年1月5日 — 2月22日,采用完全随机抽样的方法,采集安庆市38家农贸市场、4例确诊人感染H7N9禽流感病例居家禽类相关标本共324份,用实时荧光定量RT-PCR方法进行甲型流感病毒通用核酸检测,对阳性标本进一步进行H7N9检测。  结果  安庆市H7N9禽流感病毒阳性率28.40 %,11个县（市、区）中有10个检出H7N9病毒阳性,阳性率90.90 % （10/11）。主城区阳性率37.68 %（26/69）高于非主城区,差异有统计学意义（χ² = 9.23,P < 0.05）;鸡、鸭、鸽子标本中,鸡标本阳性率最高31.75 %（80/252）,差异有统计学意义（χ² = 6.57,P < 0.05）;农贸市场来源的标本阳性率为29.77 %（92/309）,病例家来源的标本阳性率为0,差异有统计学意义（χ² = 6.24,P < 0.05）;不同类型标本中,咽拭子和肛拭子阳性率最高,分别为42.86 %（9/21）和40.00 %（2/5）,其次为饮水和污水;发生人感染病例前外环境标本阳性率38.71 %（12/31）高于发生后,但2者间的差异无统计学意义（χ² = 1.79,P > 0.05）。  结论  安庆市农贸市场存在H7N9禽流感病毒污染。     

An epidemiological study of influenza viruses among Chinese farm families with household ducks and pigs.

To examine the possibility of interspecies transmission and genetic reassortment of influenza viruses on farms in Southern China, we surveyed 20 farm families living outside the city of Nanchang who raised pigs and ducks in their homes. Weekly interviews of family members and virus isolation studies of throat swabs and faecal samples, collected from September 1992 to September 1993, established the seasonal pattern of respiratory tract infections in these families and identified 11 influenza viruses (6 in humans and 5 in ducks). Most of the human isolates were type A of H3N2 subtype. Serologic studies of farm pigs indicated infection by the same human viruses circulating in family members, but there was no evidence that either swine or avian viruses had been transmitted to pigs. Eight of 156 human serum samples inhibited the neuraminidase activity of two of the duck isolates, raising the possibility of interspecies transmission of these avian viruses. Genotype analysis of duck and human isolates provided no evidence for reassortment. Our finding support the concept that intermingling of humans, pigs and ducks on Chinese farms is favourable to the generation of new, potentially hazardous strains of influenza virus.     

Identification of the source of A (H10N8) virus causing human infection

A novel H10N8 influenza A virus has been detected in three humans in China since December 2013. Although this virus was hypothesized to be a novel reassortant among influenza viruses from wild birds and domestic poultry, its evolutionary path leading to human infection is unknown. Sporadic surveillance at the live poultry market (LPM) suspected to be the source of infection for the first H10N8 patient has shown a gradual increase in influenza virus prevalence culminating with a predominance of H10N8 viruses. Influenza viruses detected in the LPM up to 8 months prior to human infection contributed genetic components to the zoonotic virus. These H10N8 viruses have continued to evolve within this LPM subsequent to the human infection, and continuous assessments of these H10N8 viruses will be necessary. Serological surveillance showed that the virus appears to have been present throughout the LPM system in Nanchang, China. Reduction of the influenza virus burden in LPMs is essential in preventing future emergence of novel influenza viruses with zoonotic and pandemic potential.     

2011 - 2015年宿迁市禽流感职业暴露人群和环境监测结果分析

摘要:目的 了解宿迁市职业暴露人群H5N1 禽流感病毒抗体水平和环境中禽流感病毒的分布情况,为禽流感防控提供科学依据。方法 在2011 - 2015年采集从事家禽养殖、屠宰等人群血清标本用血凝抑制试验检测H5N1 抗体;对在城乡活禽市场等采集的环境标本用Real-time PCR法检测禽流感病毒FluA、H5、H7、H9核酸。结果 职业暴露人群572人份血清A（H5N1）抗体均为阴性;1 038份环境标本共检出流感A型阳性标本65份,阳性率为6.26%（其中H5阳性率1.64%,H7阳性率0.19%,H9阳性率3.28%,A未分型0.48%,混合感染0.58%）。结论 职业暴露人群血清标本中未检测出A（H5N1）禽流感病毒抗体阳性标本;宿迁市监测点环境中存在H5、H7、H9 禽流感病毒,存在人感染高致病性禽流感的风险。禽流感感染风险主要集中在城乡活禽市场。     

M2SR,a novel live influenza vaccine,protects mice and ferrets against highly pathogenic avian influenza

The emergence of highly pathogenic avian influenza H5N1 viruses has heightened global concern about the threat posed by pandemic influenza. To address the need for a highly effective universal influenza vaccine, we developed a novel M2-deficient single replication (M2SR) influenza vaccine virus and previously reported that it provided strong heterosubtypic protection against seasonal influenza viruses in mice. In the current study, we assessed M2SR induced protection against H5N1 influenza in mice and ferrets.Mice were intranasally inoculated with M2SR viruses containing the HA and NA from A/Vietnam/1203/2004 (M2SR H5N1) or A/California/07/2009 (M2SR H1N1). All M2SR vaccinated mice survived lethal challenge with influenza A/Vietnam/1203/2004 (H5N1), whereas 40% of mice vaccinated with recombinant H5 HA and none of the naïve controls survived. M2SR H5N1 provided sterile immunity, whereas low levels of virus were detected in the lungs of some M2SR H1N1 vaccinated mice. In contrast, recombinant H5 HA vaccinated mice and naïve controls showed systemic infection.M2SR H5N1 induced strong serum and mucosal antibody responses (IgG and IgA classes) against H5 HA, with high hemagglutination inhibition (HAI) titers. In contrast, while M2SR H1N1 elicited cross-reactive antibodies recognizing the H5 HA2 stalk region or the neuraminidase, no HAI activity against H5N1 virus was detected after M2SR H1N1 immunization.Both M2SR H5N1 and H1N1 also protected ferrets against lethal challenge with A/Vietnam/1203/2004. A prime–boost regimen provided optimal protection with no virus detected in the respiratory tract or brain after challenge. As in the mouse model, only the M2SR H5N1 vaccine induced HAI antibodies against the challenge virus in ferrets, while the M2SR H1N1 was able to provide protection without the induction of HAI antibodies.In summary, effective protection against highly pathogenic H5N1 influenza virus was provided by both homologous H5N1 M2SR and heterologous H1N1 M2SR demonstrating the cross-protective attributes of the M2SR platform.     

Molecular evolution of H5N1 in Thailand between 2004 and 2008

Highly pathogenic avian influenza (HPAI) H5N1 viruses have seriously affected the Asian poultry industry since their occurrence in 2004. Thailand has been one of those countries exposed to HPAI H5N1 outbreaks. This project was designed to compare the molecular evolution of HPAI H5N1 in Thailand between 2004 and 2008. Viruses with clade 1 hemagglutinin (HA) were first observed in early 2004 and persisted until 2008. Viruses with clade 2.3.4 HA were first observed in the northeastern region of Thailand between 2006 and 2007. Phylogenetic analysis among Thai isolates indicated that clade 1 viruses in Thailand consist of three distinct lineages: CUK2-like, PC168-like, and PC170-like viruses. The CUK2-like virus represents the predominant lineage and has been circulating throughout the course of the 4-year outbreaks. Analysis of recently isolated viruses has shown that the genetic distance was slightly different from viruses of the early outbreak and that CUK2-like viruses comprise the native strain. Between 2005 and 2007, PC168-like and PC170-like viruses were first observed in several areas around central and lower northern Thailand. In 2008, viruses reassorted from these two lineages, PC168-like and PC170-like viruses, were initially isolated in the lower northern provinces of Thailand and subsequently spread to the upper central part of Thailand. On the other hand, CUK2-like viruses were still detected around the lower northern and the upper central part of Thailand. Furthermore, upon emergence of the reassorted viruses, the PC168-like and PC170-like lineages could not be detected, suggesting that the only predominant strains still circulating in Thailand were CUK2-like and reassorted viruses. The substitution rate among clade 1 viruses in Thailand was lower. The virus being limited to the same area might explain the lower nucleotide substitution rate. This study has demonstrated that nationwide attempts to monitor the virus may help curb access and propagation of new HPAI viral genes.     

济宁市禽流感职业暴露人群血清学和环境禽流感 监测分析

摘要:目的　了解济宁市职业暴露人群禽流感病毒的接触感染状况和环境禽流感病毒分布情况。方法　2010-2014年采集禽流感职业暴露人群的血清标本,采用马红细胞血凝抑制试验检测职业暴露人群H5N1血凝素抗体,采集环境标本采用Real-time PCR对标本进行A型及H5、H7、H9亚型禽流感病毒核酸检测。结果　本研究共采集677份家禽规模养殖场等禽流感职业暴露人群血清标本,进行H5N1抗体检测结果均为阴性;共采集城乡活禽市场等各类监测点的水、禽类（含候鸟）粪便及笼具表面等环境标本1754份,经核酸检测A型阳性28份（1.60%）,H9亚型阳性10份（35.71%）,H5亚型阳性1份（3.57%）,A未分型15份（53.57%）,2份样本H9、H5均为阳性,未检出H7亚型。仅从城乡活禽市场和规模养殖场检测到阳性标本,野生候鸟栖息地及其他场所未检测出阳性标本。粪便等不同类型标本中均检出阳性标本。结论　济宁市城乡活禽市场和规模养殖场中有H5、H9禽流感病毒存在,城乡活禽市场是禽流感病毒重要的存在场所,存在人感染禽流感风险,应加强禽流感监测并做好综合性防控措施。     

基于H5Nl证据基础的人禽流感流行病学特征

该文基于近年来国内外报道的高致病性禽流感病毒H5N1流行病学客观证据,借鉴循证医学的思路,系统综述H5N1病毒特征、感染源的广泛性、传播途径的隐蔽性和易感人群的可获得性等流行特征,阐述人禽流感暴发流行的可能性及预防控制的紧迫性。