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.     

Qualitative risk assessment of transmission pathways of highly pathogenic avian influenza (HPAI) virus at live poultry markets in Dhaka city,Bangladesh

Analysis of environmental samples obtained from the Live Poultry Markets (LPMs) of Dhaka City, Bangladesh, has revealed that the highest degree of prevalence of highly pathogenic avian influenza A (HPAI, H5N1), besides other subtypes of the LPAI virus, poses the plausible risk of transmission of these viruses between human and poultry species. The present study was conducted using the OIE risk analysis framework to assess the risk level of each pathway successively. The estimated risk parameters were integrated towards to obtain the overall risk level for each specific HPAI transmission pathway using the matrix adapted by Cristobel Zepeda accompanying other expert consultations. The relevant data obtained from published and unpublished sources, together with survey data of field observations, were used to formulate and confirm the risk pathways and their associated risks. The results revealed that the risk of the release of the HPAI virus was medium when exposure was high. Additionally, the consequence would be considered very high with a medium degree of uncertainty for all parameters. Ultimately, the overall risk for transmission was estimated as medium with a medium degree of uncertainty. The findings of this study reveal that there is a significant threat that HPAI virus transmission could occur among poultry and humans and effectively sustain within the environment of the LPMs. Our findings are primarily focused on public health considerations, the hygienic slaughter of poultry and the relevant cleaning and sanitation practices conducted in the LPMs to support evidence‐based decision‐making processes. The findings of the study have the potential to be used to formulate effective risk reduction measures and can be further adapted in low‐resource settings without major infrastructural changes required of the LPMs. All of which would reduce the risk of HPAI virus release and further lessen the degree of exposure and transmission in established LPMs.     

Evaluation of effectiveness and efficiency of wild bird surveillance for avian influenza

This study aimed to assess which method of wild waterbird surveillance had the greatest probability of detecting highly pathogenic avian influenza (HPAI) H5N1 during a period of surveillance activity, the cost of each method was also considered. Lake Constance is a major wintering centre for migratory waterbirds and in 2006 it was the site of an HPAI H5N1 epidemic in wild birds. Avian influenza surveillance was conducted using harmonised approaches in the three countries around the lake, Austria, Germany and Switzerland, from 2006–2009. The surveillance consisted of testing birds sampled by the following methods: live birds caught in traps, birds killed by hunters, birds caught in fishing nets, dead birds found by the public and catching live Mute Swans (Cygnus olor); sentinel flocks of Mallards (Anas platyrhynchos) were also used. Scenario tree analysis was performed including sensitivity analysis, followed by assessment of cost-effectiveness. Results indicated that if HPAI H5N1 was present at 1% prevalence and assuming HPAI resulted in bird mortality, sampling dead birds found by the public and sentinel surveillance were the most sensitive approaches despite residual uncertainty over some parameters. The uncertainty over the mortality of infected birds was an influential factor. Sampling birds found dead was most cost-effective, but strongly dependent on mortality and awareness of the public. Trapping live birds was least cost-effective. Based on our results, we recommend that future HPAI H5N1 surveillance around Lake Constance should prioritise sentinel surveillance and, if high mortality is expected, the testing of birds found dead.     

Predictors for Risk Factors for Spread of Avian Influenza Viruses by Poultry Handlers in Live bird markets in Uganda

Live bird markets (LBMs) are essential for marketing poultry, but have been linked to many outbreaks of avian influenza (AI) and its spread. In Uganda, it has been observed that demographic characteristics of poultry traders/handlers influence activities and decision‐making in LBMs. The study investigated the influence of socio‐demographic characteristics of poultry handlers: age, sex, religion, educational background, level of income, location of residence and region of operation on 20 potential risk factors for introduction and spread of AI in LBMs. Study sites included 39 LBMs in the four regions of Uganda. Data was collected using a semi‐structured questionnaire administered to 424 poultry handlers. We observed that background of education was a predictor for slaughter and processing of poultry in open sites. Location of residence was associated with slaughter of poultry from open sites and selling of other livestock species. Region influenced stacking of cages, inadequate cleaning of cages, feeders and drinkers, and provision of dirty feed and water. Specifically, bird handlers with secondary level of education (OR = 12.9, 95% CI: 2.88–57.4, P < 0.01) were more likely to be involved in open site slaughter of poultry than their counterparts without formal education. Comparatively, urbanite bird handlers were less likely to share poultry equipment (OR = 0.4, 95% CI: 0.22–0.63, P < 0.01) than rural resident handlers. Poultry handlers in Northern were 3.5 times more likely to practise insufficient cleaning of cages (OR = 3.5, 95% CI: 1.52–8.09) compared to those in Central region. We demonstrated that some socio‐demographic characteristics of poultry handlers were predictors to risky practices for introduction and spread of AI viruses in LBMs in Uganda.     

吉林省长春市城乡活禽市场H9N2亚型禽流感病毒的检测与分子遗传进化分析

H9N2亚型禽流感病毒(Avian influenza virus, AIV)通过为其他流感病毒提供内部基因或直接跨越种间屏障感染人,而活禽市场是H9N2亚型AIV传播的主要传播途径之一。为了解吉林省长春地区城乡活禽市场H9N2亚型AIV流行特点和趋势,对2021年9月至2023年4月在4个城乡活禽市场分离到3株代表性H9N2亚型AIV进行了分子遗传进化分析。结果显示3株毒株HA蛋白裂解位点均为PSKSSR↓GLF,其受体结合位点的第226位氨基酸由Q突变为L,可与α,2-6唾液酸受体结合,具有感染人的特性。分子遗传进化分析显示3株毒株的HA基因归属于BJ-94谱系中h9.4.2.5亚分支;NA基因归属Y280谱系;PB2、M基因均属于G1谱系;剩余内部基因均属于F-98谱系。研究结论丰富当前国内H9N2亚型AIV的流行病学研究,提示需加强病毒变异监控和新疫苗研发。     

Surveillance for highly pathogenic avian influenza in wild birds in the USA

As part of the USA's National Strategy for Pandemic Influenza, an Interagency Strategic Plan for the Early Detection of Highly Pathogenic H5N1 Avian Influenza in Wild Migratory Birds was developed and implemented. From 1 April 2006 through 31 March 2009, 261 946 samples from wild birds and 101 457 wild bird fecal samples were collected in the USA; no highly pathogenic avian influenza was detected. The United States Department of Agriculture, and state and tribal cooperators accounted for 213 115 (81%) of the wild bird samples collected; 31, 27, 21 and 21% of the samples were collected from the Atlantic, Pacific, Central and Mississippi flyways, respectively. More than 250 species of wild birds in all 50 states were sampled. The majority of wild birds (86%) were dabbling ducks, geese, swans and shorebirds. The apparent prevalence of low pathogenic avian influenza viruses during biological years 2007 and 2008 was 9.7 and 11.0%, respectively. The apparent prevalence of H5 and H7 subtypes across all species sampled were 0.5 and 0.06%, respectively. The pooled fecal samples (n= 101 539) positive for low pathogenic avian influenza were 4.0, 6.7 and 4.7% for biological years 2006, 2007 and 2008, respectively. The highly pathogenic early detection system for wild birds developed and implemented in the USA represents the largest coordinated wildlife disease surveillance system ever conducted. This effort provided evidence that wild birds in the USA were free of highly pathogenic avian influenza virus (given the expected minimum prevalence of 0.001%) at the 99.9% confidence level during the surveillance period.     

2009—2010年华东地区家禽低致病性禽流感病毒的流行病学调查与分析

为了解华东地区家禽中低致病性禽流感病毒（low pathogenic avian influenza viruses,LPAIVs）的分布规律,从2009年10月到2010年9月在华东地区某活禽市场采集鸡、鸭、鹅等家禽的泄殖腔拭子共1 650份,经鸡胚接种和HA、HI试验鉴定,结果从58份样品中分离到了LPAIVs,总分离率为3.51%。所分离到的6种HA亚型及各HA亚型分离率从高到底依次为：H6、H3、H1、H4、H9、H11。从这些样品中鉴定出7种NA亚型,包括N1、N2、N3、N4、N5、N6、N8,二者之间有11种组合。家鸭样品中LPAIVs的分离率为7.28%,显著高于鸡源样品的分离率1.00%和鹅源样品的分离率1.02%。LPAIVs的季节性分布较为明显,3～6月份和10～12月份的分离率较高,而冬季最冷的1月份和夏季最热的7月份则没有分离到。2种或2种以上不同HA亚型混合感染的样品有6份,全部为水禽源样品,占总阳性样品数的10.34%。这些数据表明活禽市场可以作为AIV的一个重要储存库,而家养水禽可作为AIV的一个重要储存宿主,应该继续加强对活禽市场,尤其是家养水禽中AIV的监测。     

Serological evidence of avian influenza virus subtype H5 and H9 in live bird market,Myanmar

Avian Influenza (AI), caused by Alphainfluenzaviruses (AIVs), is a contagious respiratory disease in birds and mammals. AIVs have been reported in poultry worldwide and the impact of AIVs on human health is immense. In this study, a serological survey of AIV subtype H5 and H9 was conducted in a live bird market (LBM) in Yangon, Myanmar during February 2016 to September 2016. A total of 621 serum samples were collected from chickens (n = 489) and ducks (n = 132) from 48 vendors in the LBM. The samples were examined for antibodies against influenza viruses by using NP-ELISA and specific antibodies against AIV-H5N1 (Clade 2.3.4) and AIV-H9N2 (Clade 9.4.2) by using Hemagglutination Inhibition (HI) assay. The result of NP-ELISA assay showed that 12.88 % (80/621) of poultry in LBM was positive for AIV antibodies. In detail, 38.06 % (51/134) of layers, 7.08 % (8/113) of backyard chicken, 2.07 % (5/242) of broilers and 12.12 % (16/132) of ducks were AIV positive. The HI test for specific antibodies against AIV-H5N1 and AIV-H9N2 were 1.77 % (11/621) and 4.51 % (28/621), respectively. Our findings revealed the evidence of AIV-H5N1 and AIV-H9N2 exposure in both chicken and ducks in the LBM in Yangon, Myanmar. Risks of influenza infections and transmission among poultry and humans in the LBMs could not be ignored.     

香港活禽市场的管理措施

香港于1997年发生全球首例人感染H5N1禽流感事件,随后几年又陆续从当地活禽市场分离到H5N1高致病性禽流感病毒。为降低人和家禽感染H5N1等高致病性禽流感病毒的风险,香港根据相关的研究和调查,对活禽市场陆续采取了一系列针对性措施,并对所采取的管理措施进行了科学的评估,有效地控制了高致病性禽流感疫情。笔者对香港活禽市场的管理措施、实施的技术依据、实施后的效果评估等方面进行了综述,以期为内地调整活禽市场管理措施,降低人和家禽通过其感染H5N1等高致病性禽流感的风险提供参考和借鉴。     

The characterization of low pathogenic avian influenza viruses isolated from wild birds in northern Vietnam from 2006 to 2009

Due to concerns that wild birds could possibly spread H5N1 viruses, surveillance was conducted to monitor the types of avian influenza viruses circulating among the wild birds migrating to or inhabiting in northern Vietnam from 2006 to 2009. An H5N2 virus isolated from a Eurasian woodcock had a close phylogenetic relationship to H5 viruses recently isolated in South Korea and Japan, suggesting that H5N2 has been shared between Vietnam, South Korea, and Japan. An H9N2 virus isolated from a Chinese Hwamei was closely related to two H9N2 viruses that were isolated from humans in Hong Kong in 2009, suggesting that an H9N2 strain relevant to the human isolates had been transmitted to and maintained among the wild bird population in Vietnam and South China. The results support the idea that wild bird species play a significant role in the spread and maintenance of avian influenza and that this also occurs in Vietnam.     

辽宁省低致病性禽流感（H9）带毒情况调查

为确实掌握辽宁省家禽低致病性禽流感（H9）带毒或感染情况,2011～2012年,辽宁省动物疫病预防控制中心对全省共64个禽场（户）进行低致病性禽流感带毒或感染情况调查。结果表明,近7成以上的场（户）未进行H9亚型禽流感免疫。在所监测的5例H9亚型禽流感阳性禽群中,除1例免疫情况不明外,其他均未进行免疫,阳性场比率有逐年升高之势,其危害应引起高度重视。     

An inactivated vaccine to control the current H9N2 low pathogenic avian influenza in Korea

The H9N2 subtype low pathogenic avian influenza is one of the most prevalent avian diseases worldwide, and was first documented in 1996 in Korea. This disease caused serious economic loss in Korea''s poultry industry.In order to develop an oil-based inactivated vaccine, a virus that had been isolated in 2001 (A/chicken/Korea/01310/2001) was selected based on its pathogenic, antigenic, and genetic properties. However, in animal experiments, the efficacy of the vaccine was found to be very low without concentration of the antigen (2⁷ to 2¹⁰ hemagglutinin unit). In order to overcome the low productivity, we passaged the vaccine candidate virus to chicken eggs. After the 20th passage, the virus was approximately ten times more productive compared with the parent virus. For the most part, the passaged virus maintained the hemagglutinin cleavage site amino acid motif (PATSGR/GLF) and had only three amino acid changes (T133N, V216G, E439D, H3 numbering) in the hemagglutinin molecule, as well as 18 amino acid deletions (55-72) and one amino acid change (E54D) in the NA stalk region. The amino acid changes did not significantly affect the antigenicity of the vaccine virus when tested by hemagglutination inhibition assay. Though not complete, the vaccine produced after the 20th passage of the virus (01310 CE20) showed good protection against a homologous and recent Korean isolate (A/chicken/Korea/Q30/2004) in specific pathogen- free chickens.The vaccine developed in this study would be helpful for controlling the H9N2 LPAI in Korea.     

禽流感免疫程序的建立

通过禽流感病毒保护性HI抗体滴度、禽流感母源抗体消长规律以及疫苗免疫抗体消长规律的研究,结合疫苗和诊断试剂质量及血清监测中存在的问题制定了禽流感免疫程序。生产中应用表明,本研究制定的免疫程序是切实可行的。     

Molecular characterization of low pathogenic avian influenza viruses, isolated from food products imported into Singapore

We have completed the genetic characterization of all eight gene segments for four low pathogenic avian influenza (LPAI) viruses. The objective of this study was to detect the presence of novel signatures that may serve as early warning indicators of the conversion of LPAI viruses to high pathogenic avian influenza (HPAI) viruses. This study included three H5N2 and one H5N3 viruses that were isolated from live poultry imported into Singapore as part of the national avian influenza virus (AIV) surveillance program. Based on the molecular criterion of the World Organisation for Animal Health (OIE), sequence analysis with the translated amino acid (aa) sequence of the hemagglutinin (HA) gene revealed the absence of multibasic aa at the HA cleavage site, identifying all four virus isolates as LPAI. Detailed phylogenetic tree analyses using the HA and neuraminidase (NA) genes clustered these isolates in the Eurasian H5 lineage, but away from the HPAI H5 subtypes. This analysis further revealed that the internal genes clustered to different avian and swine subtypes, suggesting that the four isolates may possibly share their ancestry with these different influenza subtypes. Our results suggest that the four LPAI isolates in this study contained mainly avian signatures, and the phylogenetic tree for the internal genes further suggests the potential for reassortment with other different circulating avian subtypes. This is the first comprehensive report on the genetic characterization of LPAI H5N2/3 viruses isolated in South-East Asia.     

Molecular identification of the vaccine strain from the inactivated oil emulsion H9N2 low pathogenic avian influenza vaccine

In order to control the H9N2 subtype low pathogenic avian influenza (LPAI), an inactivated vaccine has been used in Korea since 2007. The Korean veterinary authority permitted the use of a single H9N2 LPAI vaccine strain to simplify the evolution of the circulating virus due to the immune pressure caused by the vaccine use. It is therefore important to determine the suitability of the vaccine strain in the final inactivated oil emulsion LPAI vaccine. In this study, we applied molecular rather than biological methods to verify the suitability of the vaccine strain used in commercial vaccines and successfully identified the strain by comparing the nucleotide sequences of the hemagglutinin and neuraminidase genes with that of the permitted Korean LPAI vaccine strain. It is thought that the method used in this study might be successfully applied to other viral genes of the LPAI vaccine strain and perhaps to other veterinary oil emulsion vaccines.     

Surveillance for highly pathogenic avian influenza in migratory shorebirds at the terminus of the East Asian-Australasian Flyway

AIM: To determine if migratory birds arriving in New Zealand in the Southern Hemisphere spring of 2004 were infected with the highly pathogenic avian influenza (AI) virus, H5N1.

METHODS: Cloacal and faecal samples were collected from migratory red knots following their arrival in New Zealand in October 2004. Two species of resident sympatric birds, wrybill and mallard duck, were sampled prior to, and following, the arrival of migratory birds.

RESULTS: No AI viruses were isolated from migratory or resident shorebirds. Non-pathogenic AI viruses were isolated from six resident mallard ducks, comprising the endemic subtypes H4 (n=2), H7 (non-pathogenic), H10, and H11 (n=2).

CONCLUSIONS: Highly pathogenic AI H5N1 virus was not detected in migratory shorebirds or sympatric water birds in the Firth of Thames, New Zealand, in 2004-2005, despite the possible proximity of migratory birds to outbreaks of the disease in East Asia in 2004.     

Genetic characterization of H9N2 avian influenza virus previously unrecognized in Korea

In this study, we describe the isolation and characterization of previously unreported Y280-lineage H9N2 viruses from two live bird markets in Korea in June 2020. Genetic analysis revealed that they were distinct from previous H9N2 viruses circulating in Korea and had highest homology to A/chicken/Shandong/1844/2019(H9N2) viruses. Their genetic constellation showed they belonged to genotype S, which is the predominant genotype in China since 2010, where genotype S viruses have infected humans and acted as internal gene donors to H5 and H7 zoonotic influenza viruses. Active surveillance and control measures need to be enhanced to protect the poultry industry and public health.