全球高致病性禽流感风险评估

H5N1高致病性禽流感暴发的风险管理需要对不同的风险因素进行综合评估,这些因素影响不同地理区域传染病的传播和持续时间。禽流感灾害风险评估的关键是制定风险路径,然后用它来发展一种定性或定量的风险评估模型。本文建立在风险分析模型框架基础之上,利用FAO、WHO以及OIE等国际组织和有关国家政府可公开获得的数据和信息,利用ArcGIS对H5N1高致病性禽流感进行风险评估,并对野生鸟类和家禽贸易在H5N1高致病性禽流感传播中的重要性进行分析。     

Risk factors and characteristics of H5N1 Highly Pathogenic Avian Influenza (HPAI) post-vaccination outbreaks

Highly pathogenic avian influenza (HPAI) virus H5N1 is now endemic in South-East Asia but HPAI control methods differ between countries. A widespread HPAI vaccination campaign that started at the end of 2005 in Viet Nam resulted in the cessation of poultry and human cases, but in 2006/2007 severe HPAI outbreaks re-emerged. In this study we investigated the pattern of this first post-vaccination epidemic in southern Viet Nam identifying a spatio-temporal cluster of outbreak occurrence and estimating spatially smoothed incidence rates of HPAI. Spatial risk factors associated with HPAI occurrence were identified. Medium-level poultry density resulted in an increased outbreak risk (Odds ratio (OR) = 5.4, 95% confidence interval (CI): 1.6–18.9) but also climate-vegetation factors played an important role: medium-level normalised difference vegetation indices during the rainy season from May to October were associated with higher risk of HPAI outbreaks (OR = 3.7, 95% CI: 1.7–8.1), probably because temporal flooding might have provided suitable conditions for the re-emergence of HPAI by expanding the virus distribution in the environment and by enlarging areas of possible contacts between domestic waterfowl and wild birds. On the other hand, several agricultural production factors, such as sweet potatoes yield, increased buffalo density, as well as increased electricity supply were associated with decreased risk of HPAI outbreaks. This illustrates that preventive control measures for HPAI should include a promotion of low-risk agricultural management practices as well as improvement of the infrastructure in village households. Improved HPAI vaccination efforts and coverage should focus on medium poultry density areas and on the pre-monsoon time period.     

辽宁省一起野禽H5N6高致病性禽流感疫情的紧急流行病学调查

2021年4月6日,辽宁省沈阳市和平区长白岛森林公园内饲养的10只黑天鹅和1只大雁死亡,袭击率为3.8％(11/291).经现场调查和临床剖检诊断,市级和省级疫病控制中心实验室检测,诊断为疑似H5亚型高致病性禽流感.随后,经国家禽流感参考实验室确诊为H5N6亚型高致病性禽流感.4月12日,农业农村部公布了该起疫情.在接...     

Highly pathogenic avian influenza virus subtype H5N1 in ducks in the Northern part of Cameroon

Highly pathogenic avian influenza (HPAI) virus was first detected in Cameroon in February 2006. Analysis of NA sequences of the virus demonstrated that it is closely related to the H5N1 isolates from Northern Nigeria, Sudan and Ivory Coast, suggesting a common virus ancestor.     

A single dose of inactivated oil-emulsion bivalent H5N8/H5N1 vaccine protects chickens against the lethal challenge of both highly pathogenic avian influenza viruses

In this study, two highly pathogenic avian influenza (HPAI) H5N8 viruses were isolated from chicken and geese in 2018 and 2019 (Chicken/ME-2018 and Geese/Egypt/MG4/2019). The hemagglutinin and neuraminidase gene analyses revealed their close relatedness to the clade-2.3.4.4b H5N8 viruses isolated from Egypt and Eurasian countries. A monovalent inactivated oil-emulsion vaccine containing a reassortant virus with HA gene of the Chicken/ME-2018/H5N8 strain and a bivalent vaccine containing same reassortant virus plus a previously generated reassortant H5N1 strain (CK/Eg/RG-173CAL/17). The safety of both vaccines was evaluated in specific-pathogen-free (SPF) chickens. To evaluate the efficacy of the prepared vaccines, 2-week-old SPF chickens were vaccinated with 0.5 mL of a vaccine formula containing 10⁸/EID₅₀ /dose from each strain via the subcutaneous route. Vaccinated birds were challenged with either wild-type HPAI-H5N8 or H5N1 viruses separately at 3 weeks post-vaccine. Results revealed that both vaccines induced protective hemagglutination-inhibiting (HI) antibody titers as early as 2 weeks PV (≥5.0 log₂). Vaccinated birds were protected clinically against both subtypes (100 % protection). HPAI-H5N1 virus shedding was significantly reduced in birds that were vaccinated with the bivalent vaccine; meanwhile, HPAI-H5N8 virus shedding was completely neutralized in both tracheal and cloacal swabs after 3 days post-infection in birds that had been vaccinated with either vaccine. In conclusion, the developed bivalent vaccine proved to be efficient in protecting chickens clinically and reduced virus shedding via the respiratory and digestive tracts. The applicability of the multivalent avian influenza vaccines further supported their value to facilitate vaccination programs in endemic countries.     

Experimental infection of chickens, ducks and quails with the highly pathogenic H5N1 avian influenza virus

Highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype have spread since 2003 in poultry and wild birds in Asia, Europe and Africa. In Korea, the highly pathogenic H5N1 avian influenza outbreaks took place in 2003/2004, 2006/2007 and 2008. As the 2006/2007 isolates differ phylogenetically from the 2003/2004 isolates, we assessed the clinical responses of chickens, ducks and quails to intranasal inoculation of the 2006/2007 index case virus, A/chicken/Korea/IS/06. All the chickens and quails died on 3 days and 3-6 days post-inoculation (DPI), respectively, whilst the ducks only showed signs of mild depression. The uninoculated chickens and quails placed soon after with the inoculated flock died on 5.3 and 7.5 DPI, respectively. Both oropharyngeal and cloacal swabs were taken for all three species during various time intervals after inoculation. It was found that oropharyngeal swabs showed higher viral titers than in cloacal swabs applicable to all three avian species. The chickens and quails shed the virus until they died (up to 3 to 6 days after inoculation, respectively) whilst the ducks shed the virus on 2-4 DPI. The postmortem tissues collected from the chickens and quails on day 3 and days 4-5 and from clinically normal ducks that were euthanized on day 4 contained the virus. However, the ducks had significantly lower viral titers than the chickens or quails. Thus, the three avian species varied significantly in their clinical signs, mortality, tissue virus titers, and duration of virus shedding. Our observations suggest that duck and quail farms should be monitored particularly closely for the presence of HPAIV so that further virus transmission to other avian or mammalian hosts can be prevented.     

Assessing the risk of highly pathogenic avian influenza H5N1 transmission through poultry movements in Bali,Indonesia

Indonesia continues to report the highest number of human and poultry cases of highly pathogenic avian influenza H5N1. The disease is considered to be endemic on the island of Bali. Live bird markets are integral in the poultry supply chain on Bali and are important, nutritionally and culturally, for the rural and urban human populations. Due to the lack of biosecurity practiced along the supply chain from producer to live bird markets, there is a need to understand the risks associated with the spread of H5N1 through live bird movements for effective control. Resources to control H5N1 in Indonesia are very limited and cost effective strategies are needed. We assessed the probability a live bird market is infected through live poultry movements and assessed the effects of implementing two simple and low cost control measures on this risk. Results suggest there is a high risk a live bird market is infected (0.78), and risk mitigation strategies such as detecting and removing infected poultry from markets reduce this risk somewhat (range 0.67–0.76). The study demonstrates the key role live poultry movements play in transmitting H5N1 and the need to implement a variety of control measures to reduce disease spread.     

山东省首起野生天鹅H5N8高致病性禽流感疫情的紧急调查及处置

2021年1月12—19日,山东黄河三角洲国家级自然保护区大汶流管理站出现35只死亡的野生疣鼻天鹅.经现场调查,市级和省级实验室检测,诊断为疑似H5N8亚型高致病性禽流感,经国家禽流感参考实验室进行病毒分离鉴定,最终确定为H5N8亚型高致病性禽流感疫情.疫情发生后,通过现场调查、座谈及实验室检测等方式,对此次疫情开展了...     

Infectious dose‐dependent accumulation of live highly pathogenic avian influenza H5N1 virus in chicken skeletal muscle—implications for public health

Highly pathogenic avian influenza viruses (HPAIV) of H5N1 subtype are a major global threat to poultry and public health. Export of poultry products, such as chicken and duck meat, is a known source for the cross‐boundary spread of HPAI H5N1 viruses. Humans get infected with HPAI H5N1 viruses either by close contact with infected poultry or through consumption of fresh/undercooked poultry meat. Skeletal muscle is the largest soft tissue in chicken that has been shown to contain virus during systemic HPAIV infection and supports productive virus infection. However, the time between infection of a chicken with H5N1 virus and presence of virus in muscle tissue is not yet known. Further, it is also not clear whether chicken infected with low doses of H5N1 virus that cause non‐fatal subclinical infections continue to accumulate virus in skeletal muscle. We investigated the amount and duration of virus detection in skeletal muscle of chicken experimentally infected with different doses (10², 10³ and 10⁴ EID₅₀) of a HPAI H5N1 virus. Influenza viral antigen could be detected as early as 6 hr after infection and live virus was recovered from 48 hr after infection. Notably, chicken infected with lower levels of HPAI H5N1 virus (i.e., 10² EID₅₀) did not die acutely, but continued to accumulate high levels of H5N1 virus in skeletal muscle until 6 days post‐infection. Our data suggest that there is a potential risk of human exposure to H5N1 virus through meat from clinically healthy chicken infected with a low dose of virus. Our results highlight the need to implement rigorous monitoring systems to screen poultry meat from H5N1 endemic countries to limit the global spread of H5N1 viruses.     

Serological Surveillance of Influenza A Virus Infection in Swine Populations in Fujian Province,China: No Evidence of Naturally Occurring H5N1 Infection in Pigs

Several highly pathogenic H5N1 avian influenza viruses were isolated from swine populations in Fujian Province, China, since 2001. Because it is thought that H5N1 infection in pigs might result in virus adaptation to humans, we surveyed swine populations in Fujian Province in 2004 and 2007 for serological evidence of the infection. Twenty‐five pig farms covering all nine administrative districts of Fujian Province were sampled and a total of 1407 serum specimens were collected. The haemagglutination inhibition (HI) tests revealed no evidence of H5 infection and only a few cases of H9 infection. The negative results for H5 infection were further verified by micro‐neutralization tests. By contrast, H1 influenza virus infections were prevalent in swine in both surveys according to the results of enzyme‐linked immunosorbent assay (ELISA). The H3 infection rate was reduced dramatically in 2007 compared with 2004, when examined by HI and ELISA. In summary, the results imply that the swine populations in Fujian Province had not been affected greatly by the H5N1 avian influenza virus, given that there is no serological evidence that H5N1 influenza virus has infected the pig populations. The reported isolates represent only sporadic cases.     

流感病毒H5N1对野生动物的威胁

20世纪爆发的几次A型流感大流行曾经造成巨大的损失。进入21世纪以来A型流感病毒H5N1又在最近几年爆发，不仅给养禽业和人类造成巨大打击，也对野生动物产生了极大的威胁。本文通过分析H5N1型流感的疫情，提出了针对保护野生动物特别是圈养种群的措施。     

Critical control points for avian influenza A H5N1 in live bird markets in low resource settings

Live bird markets can become contaminated with and become a source of transmission for avian influenza viruses including the highly pathogenic H5N1 strain. Many countries affected by the H5N1-virus have limited resources for programs in environmental health, sanitation and disease control in live bird markets. This study proposes five critical control points (CCPs) to reduce the risk of H5N1-virus contamination in markets in low resource settings. The CCPs were developed based on three surveys conducted in Indonesia: a cross-sectional survey in 119 markets, a knowledge, attitudes and practice survey in 3 markets and a microbiological survey in 83 markets. These surveys assessed poultry workflow, market infrastructure, hygiene and regulatory practices and microbiological contamination with the H5N1-virus. The five CCPs identified were (1) reducing risk of receiving infected birds into the market, (2) reducing the risk of virus spread between different bird flocks in holding cages, (3) reducing surface contamination by isolating slaughter processes from other poultry-related processes, (4) minimizing the potential for contamination during evisceration of carcasses and (5) reducing the risk of surface contamination in the sale zone of the market. To be relevant for low resource settings, the CCPs do not necessitate large infrastructure changes. The CCPs are suited for markets that slaughter poultry and have capacity for daily disposal and removal of solid waste from the market. However, it is envisaged that the CCPs can be adapted for the development of risk-based programs in various settings.     

Comparative study of pandemic (H1N1) 2009, swine H1N1, and avian H3N2 influenza viral infections in quails

Quail has been proposed to be an intermediate host of influenza A viruses. However, information on the susceptibility and pathogenicity of pandemic H1N1 2009 (pH1N1) and swine influenza viruses in quails is limited. In this study, the pathogenicity, virus shedding, and transmission characteristics of pH1N1, swine H1N1 (swH1N1), and avian H3N2 (dkH3N2) influenza viruses in quails was examined. Three groups of 15 quails were inoculated with each virus and evaluated for clinical signs, virus shedding and transmission, pathological changes, and serological responses. None of the 75 inoculated (n = 45), contact exposed (n = 15), or negative control (n = 15) quails developed any clinical signs. In contrast to the low virus shedding titers observed from the swH1N1-inoculated quails, birds inoculated with dkH3N2 and pH1N1 shed relatively high titers of virus predominantly from the respiratory tract until 5 and 7 DPI, respectively, that were rarely transmitted to the contact quails. Gross and histopathological lesions were observed in the respiratory and intestinal tracts of quail inoculated with either pH1N1 or dkH3N2, indicating that these viruses were more pathogenic than swH1N1. Sero-conversions were detected 7 DPI in two out of five pH1N1-inoculated quails, three out of five quails inoculated with swH1N1, and four out of five swH1N1-infected contact birds. Taken together, this study demonstrated that quails were more susceptible to infection with pH1N1 and dkH3N2 than swH1N1.     

H5N1亚型禽流感病毒低剂量感染小鼠发病模型的建立

为模拟哺乳动物感染H5亚型高致病性禽流感病毒(HPAIV)的发病进程,本研究采用对哺乳动物高度致病的H5N1亚型HPAIV株A/bar-headed goose/Qinghai/3/05 (BHG/3/05),以低剂量鼻腔接种小鼠,观察发病、存活、病毒复制及组织病理损伤情况.结果显示,100.4 EID50即能够100％感染小鼠,但发病表现缓慢,死亡延迟至8d以后,存活达60％;体内病毒复制可持续10 d以上,感染后前3d病毒的增殖限于呼吸道,随后扩散至脑、脾、肾等其他器官;组织病理学观察肺脏早期表现出渗出性炎症,第10d发展为典型的间质性肺炎.本研究结果为探讨人禽流感的病理发生机制提供了具有价值的模型.     

Pathogenicity and transmission of H5N1 avian influenza viruses in different birds

In this study, we selected three H5N1 highly pathogenic avian influenza viruses (HPAIVs), A/Goose/Guangdong/1/1996 (clades 0), A/Duck/Guangdong/E35/2012 (clade 2.3.2.1) and A/Chicken/Henan/B30/2012 (clade 7.2) isolated from different birds in China, to investigate the pathogenicity and transmission of the viruses in terrestrial birds and waterfowl. To observe the replication and shedding of the H5N1 HPAIVs in birds, the chickens were inoculated intranasally with 10⁶ EID₅₀ of GSGD/1/96, 10³ EID₅₀ of DkE35 and CkB30, and the ducks and geese were inoculated intranasally with 10⁶ EID₅₀ of each virus. Meanwhile, the naive contact groups were set up to detect the transmission of the viruses in tested birds. Our results showed that DkE35 was highly pathogenic to chickens and geese, but not fatal to ducks. It could be detected from all the tested organs, oropharyngeal and cloacal swabs, and could transmit to the naive contact birds. GSGD/1/96 could infect chickens, ducks and geese, but only caused death in chickens. It could transmit to the chickens and ducks, but was not transmittable to geese. CkB30 was highly pathogenic to chickens, low pathogenic to ducks and not pathogenic to geese. It could be transmitted to the naive contact chickens, but not to the ducks or geese. Our findings suggested that H5N1 HPAIVs from different birds show different host ranges and tissue tropisms. Therefore, we should enhance serological and virological surveillance of H5N1 HPAIVs, and pay more attention to the pathogenic and antigenic evolution of these viruses.     

H5N1亚型高致病性禽流感病毒对鸽的致病性分析

为评估H5N1亚型禽流感病毒对鸽的致病性,用我国2004年分离的一株H5NI亚型禽流感病毒A/pigeon/GD/C2/2004( H5NI)人工感染4周龄鸽,进行了致病性试验.结果表明,该株病毒以106EID50/I00μL剂量感染能致死鸽,致死率为22.2％,病毒能在感染鸽体内广泛分布,其中肺脏和肾脏中病毒含量达到...     

我国2006年野鸟H5N1 HPAIV表面基因进化特征

国家禽流感参考实验室在2006年,从来自青、藏两省(区)3种野鸟及辽宁省的2种死亡的野鸟体内共分离到14株H5N1亚型高致病力禽流感病毒(High pathogenic avian influenza virus,HPAIV),比较发现其HA、NA基因核苷酸的同源率在97.9%～99.9%之间。所有病毒的HA基因的裂解位点均具有HPAIV特有的连续碱性氨基酸-RRRKKR-,并具有近年来H5N1亚型流行株所特有的NA基因颈部49～68位20个氨基酸缺失及NS基因80～84位5个氨基酸的缺失;遗传进化分析表明2006年野鸟病毒由2005年野鸟病毒进化而来,并形成独立的进化分支;辽宁省近2年的4株野鸟病毒亲缘关系较近,说明引起2005年该省锦州地区H5N1 HPAI疫情的病毒在该地区的野鸟体内继续存在。     

Different infection routes of avian influenza A (H5N1) virus in mice

The continuing outbreaks of avian influenza A H5N1 virus infection in Asia and Africa have caused worldwide concern because of the high mortality rates in poultry, suggesting its potential to become a pandemic influenza virus in humans. The transmission route of the virus among either the same species or different species is not yet clear. Broilers and BABL/c mice were inoculated with the H5N1 strain of influenza A virus isolated from birds. The animals were inoculated with 0.1 mL 10^6.83 TCID₅₀ of H5N1 virus oronasally, intraperitoneally and using eye drops. The viruses were examined by virological and pathological assays. In addition, to detect horizontal transmission, in each group, healthy chicks and mice were mixed with those infected. Viruses were detected in homogenates of the heart, liver, spleen, kidney and blood of the infected mice and chickens. Virus antigen was not detected in the spleen, kidney or gastrointestinal tract, but detected by Plaque Forming Unit (PFU) assay in the brain, liver and lung without degenerative change in these organs (in the group inoculated using eye drops. The detection results for mice inoculated using eye drops suggest that this virus might have a different tissue tropism from other influenza viruses mainly restricted to the respiratory tract in mice. All chicken samples tested positive for the virus, regardless of the method of inoculation. Avian influenza A H5N1 viruses are highly pathogenic to chickens, but its virulence in other animals is not yet known. To sum up, the results suggest that the virus replicates not only in different animal species but also through different routes of infection. In addition, the virus was detection not only in the respiratory tract but also in multiple extra‐respiratory tissues. This study demonstrates that H5N1 virus infection in mice can cause systemic disease and spread through potentially novel routes within and between mammalian hosts.     

H5N1亚型禽流感病毒A/goose/Guangdong/1/96的NS基因对鸡致病能力的影响

为了解H5N1亚型禽流感病毒经自然途径感染SPF鸡后，病毒的致病能力与NS基因的关系，本文主要从病理学角度比较了两株利用反向基因操作技术拯救的病毒RGSGD／1／96和RGSGD／1／2NS的致病能力。虽然只有NS基因不同，但是这两株病毒经鼻腔感染4周龄SPF鸡后表现出完全不同的致病能力，RGSGD／1／96对鸡的致死率为100％，感染鸡只的各组织脏器均可发现严重的病理损伤，该病毒在鸡体内复制能力很强，感染后3d、6d，各组织脏器均可发现大量的病毒抗原；GSGD／1／2NS对鸡的致死率为0，病毒在感染鸡体内只引起肺间质少量淋巴细胞浸润，免疫组织化学检查未发现病毒抗原信号。由于两株病毒只有NS基因不同，说明NS基因决定了H5N1亚型禽流感病毒A／goose／Guangdong／1／96对SPF鸡的致病能力。