Highly Pathogenic Avian Influenza Virus Outbreaks Research Articles

Quantitative Risk Assessment of Wind-Supported Transmission of Highly Pathogenic Avian Influenza Virus to Dutch Poultry Farms via Fecal Particles from Infected Wild Birds in the Environment.

A quantitative microbial risk assessment model was developed to estimate the probability that the aerosolization of fecal droppings from wild birds in the vicinity of poultry farms would result in the infection of indoor-housed poultry with highly pathogenic avian influenza virus (HPAIv) in the Netherlands. Model input parameters were sourced from the scientific literature and experimental data. The availability of data was diverse across input parameters, and especially parameters on the aerosolization of fecal droppings, survival of HPAIv and dispersal of aerosols were uncertain. Model results indicated that the daily probability of infection of a single poultry farm is very low, with a median value of 7.5 × 10-9. Accounting for the total number of poultry farms and the length of the bird-flu season, the median overall probability of at least one HPAIv-infected poultry farm during the bird-flu season is 2.2 × 10-3 (approximately once every 455 years). This is an overall estimate, averaged over different farm types, virus strains and wild bird species, and results indicate that uncertainty is relatively high. Based on these model results, we conclude that it is unlikely that this introduction route plays an important role in the occurrence of HPAIv outbreaks in indoor-housed poultry.

Control strategies for 2.3.4.4 highly pathogenic avian influenza virus - From vaccines to host disease resistance. Proceeding of The First International Avian Influenza Summit, University of Arkansas- October 16-17, 2023"

Avian influenza virus (AIV) is a highly contagious and lethal disease that can have major impacts on the global poultry industry and food supply. While more frequent in Asia and Europe, highly pathogenic avian influenza virus (HPAIV) outbreaks have traditionally been rare in the U.S. However, in recent years, the U.S. has seen an increase in the incidence of HPAIV outbreaks in wild birds and commercial poultry. In 2014/2015 and in 2021/2022, outbreaks of HPAIV subtype H5NX clade 2.3.4.4 resulted in the death and destruction of over 50 million birds, each costing billions of dollars to the U.S. economy. Here, we present a comprehensive overview of control strategies against the 2.3.4.4 HPAI virus, encompassing both vaccination approaches and host-driven disease resistance mechanisms. The first section outlines the advancements in vaccine development tailored specifically for the 2.3.4.4 HPAI subtype. Through a critical analysis of experimental studies and field trials, we discuss the efficacy, safety, and cross-protection capabilities of various vaccine formulations, including inactivated whole-virus vaccines, vector-based platforms, and novel recombinant technologies. Furthermore, this section explores the challenges associated with vaccine design, including strain diversity, antigenic differences, and logistical hurdles in large-scale vaccination campaigns. The second section delves into host-centric strategies aimed at enhancing natural resistance against 2.3.4.4 HPAI virus infections. Because vaccines are not currently approved for use in the U.S., control strategies for HPAIV are dependent on biosecurity and culling of infected flocks. New strategies for HPAIV control based on gene editing of poultry species could offer solutions for disease control. Two different strategies for improving disease resistance to HPAIV are based on enhancing the innate immunity of the bird and targeting the viral RNA genome inside the cell. By harnessing genetic engineering techniques, including CRISPR/Cas-9 or -13, natural resistance to avian influenza can be achieved in poultry. Taken together, we will examine the current basis of control strategies for the 2.3.4.4 HPAI virus, offering insights into the strengths and limitations of both vaccination and novel gene-editing technologies to enhance host disease resistance to HPAI. By expanding our toolbox, this work contributes to the ongoing efforts to mitigate the impact of 2.3.4.4 HPAI on global poultry industries and public health.

Can Citizen Science Contribute to Avian Influenza Surveillance?

Global change is an important driver of the increase in emerging infectious diseases in recent decades. In parallel, interest in nature has increased, and different citizen science platforms have been developed to record wildlife observations from the general public. Some of these platforms also allow registering the observations of dead or sick birds. Here, we test the utility of live, sick and dead observations of birds recorded on the platform Observation.org for the early detection of highly pathogenic avian influenza virus (HPAIV) outbreaks in the wild in Belgium and The Netherlands. There were no significant differences in the morbidity/mortality rate through Observation.org one to four weeks in advance. However, the results show that the HPAIV outbreaks officially reported by the World Organisation for Animal Health (WOAH) overlapped in time with sudden increases in the records of sick and dead birds in the wild. In addition, in two of the five main HPAIV outbreaks recorded between 2016 and 2021, wild Anseriformes mortality increased one to two months before outbreak declaration. Although we cannot exclude that this increase was related to other causes such as other infectious diseases, we propose that Observation.org is a useful nature platform to complement animal health surveillance in wild birds. We propose possible approaches to improve the utility of the platform for pathogen surveillance in wildlife and discuss the potential for HPAIV outbreak detection systems based on citizen science to complement current surveillance programs of health authorities.

Live Recombinant NDV-Vectored H5 Vaccine Protects Chickens and Domestic Ducks From Lethal Infection of the Highly Pathogenic H5N6 Avian Influenza Virus.

The H5 subtype highly pathogenic avian influenza virus (HPAIV) has been introduced to South Korea every 2 or 3 years via wild migratory waterfowls, causing devastating damages to the poultry industry. Although most damages and economic losses by HPAIV are focused on chicken layers, domestic ducks are known to play a major role in the farm-to-farm transmission. However, most HPAIV vaccine studies on poultry have been performed with oil-emulsion inactivated vaccines. In this study, we developed a live recombinant Newcastle disease virus (NDV)-vectored vaccine against H5 HPAIV (rK148/ES2-HA) using a previously established NDV vaccine strain (K148/08) isolated from a wild mallard duck. The efficacy of the vaccine when administered via the oculonasal route or as a spray was evaluated against lethal H5 HPAIV infection in domestic ducks and chickens. Oculonasal inoculation of the rK148/ES2-HA in chickens and ducks elicited antibody titers against HPAIV as early as 1 or 2 week after the single dose of vaccination, whereas spray vaccination in ducks elicited antibodies against HPAIV after the booster vaccination. The chickens and ducks vaccinated with rK148/ES2-HA showed high survival rates and low viral shedding after H5N6 HPAIV challenge. Collectively, vaccination with rK148/ES2-HA prevented lethal infection and decreased viral shedding in both chickens and ducks. The vaccine developed in this study could be useful in suppressing the viral shedding in H5 HPAIV outbreaks, with the ease of vaccine application and fast onset of immunity.

No evidence of avian influenza antibodies in two species of raptor nestlings inhabiting Norway

BackgroundSince 2016, incursions of highly pathogenic avian influenza virus (HPAIV) H5N8 clade 2.3.4.4b have caused unprecedented clinical signs and mortality in white-tailed eagles (WTE; Haliaeetus albicilla) across Europe and have been found to be infecting other raptor species, such as the northern goshawk (NG; Accipiter gentilis). Before this study, no screening of Norwegian raptors had been undertaken.ResultsPlasma samples from 43 white-tailed eagle and 29 northern goshawk nestlings, from several locations across Norway were screened for antibodies to avian influenza viruses. No antibodies, and thus, no evidence of AIV exposure, were found in these Norwegian raptors. No clinical signs of AIV were observed in 43 white tailed eagles and 29 northern goshawks.ConclusionsThere are currently no indications that white-tailed eagles and northern goshawks inhabiting Norway are threatened by the recent HPAIV outbreaks in other areas of Europe. Ongoing monitoring should, however, be maintained to detect potential future outbreaks.

Newcastle disease virus vectors expressing consensus sequence of the H7 HA protein protect broiler chickens and turkeys against highly pathogenic H7N8 virus

Continuous outbreaks of highly pathogenic avian influenza (HPAI) viruses in commercial poultry have caused devastating losses to domestic poultry with a raising public health concern. The outbreaks of HPAI viruses have increased worldwide, including the North America. Therefore, vaccination has been considered as an alternative strategy for an efficient control of HPAI viruses. In this study, we aimed to generate Newcastle disease virus (NDV) vectored H7 serotype-specific vaccines by expressing the consensus sequence of the HA protein. Conventional NDV strain LaSota vector and a chimeric NDV vector containing the avian paramyxovirus type-2 F and HN protein were able to express the consensus sequence of HA protein. The protective efficacy of vaccines was evaluated in broiler chickens and in turkeys. One-day-old poults were prime immunized with the chimeric vector expressing the HA protein followed by boost immunization with LaSota vector expressing the HA protein or co-expressing the HA and NA proteins. Our vaccine candidates provided complete protection of broiler chickens from mortality and shedding of H7N8 HPAI challenge virus. Turkeys were better protected by boosting with the LaSota vector co-expressing the HA and NA proteins than the LaSota vector expressing only the HA protein. Our study demonstrated a potential use of heterologous prime and boost vaccination strategy to protect poultry against H7 HPAI viruses.

Host Cell Mimic Polymersomes for Rapid Detection of Highly Pathogenic Influenza Virus via a Viral Fusion and Cell Entry Mechanism.

Highly pathogenic avian influenza virus (HPAIV) infections have occurred continuously and crossed the species barrier to humans, leading to fatalities. A polymerase chain reaction based molecular test is currently the most sensitive diagnostic tool for HPAIV; however, the results must be analyzed in centralized diagnosis systems by a trained individual. This requirement leads to delays in quarantine and isolation. To control the spread of HPAIV, rapid and accurate diagnostics suitable for field testing are needed, and the tests must facilitate a differential diagnosis between HPAIV and low pathogenic avian influenza virus (LPAIV), which undergo cleavage specifically by trypsin‐ or furin‐like proteases, respectively. In this study, a differential avian influenza virus rapid test kit is developed and evaluated in vitro and using clinical specimens from HPAIV H5N1‐infected animals. It is demonstrated that this rapid test kit provides highly sensitive and specific detection of HPAIV and LPAIV and is thus a useful field diagnostic tool for H5N1 HPAIV outbreaks and for rapid quarantine control of the disease.

Exploring contacts facilitating transmission of influenza A(H5N1) virus between poultry farms in West Java, Indonesia: A major role for backyard farms?

Highly pathogenic avian influenza virus (HPAIV) H5N1 has been reported in Asia, including Indonesia since 2003. Although several risk factors related to the HPAIV outbreaks in poultry in Indonesia have been identified, little is known of the contact structure of farms of different poultry production types (backyard chickens, broilers, layers, and ducks). This study aims to quantify the contact rates associated with the movement of people, and movements of live birds and products and equipment that affect the risk of HPAIV H5N1 transmission between poultry farms in Indonesia. On 124 poultry farms in 6 districts in West Java, logbooks were distributed to record the movements of farmers/staff and visitors and their poultry contacts. Most movements in backyard chicken, commercial native chicken, broiler and duck farms were visits to and from other poultry farms, whilst in layer farms visits to and from poultry companies, visits to egg collection houses and visit from other poultry farms were most frequent. Over 75% of persons visiting backyard chicken and duck farms had previously visited other poultry farms on the same day. Visitors of backyard chicken farms had the highest average contact rate, either direct contact with poultry on other farms before the visits (1.35 contact/day) or contact during their visits in the farms (10.03 contact/day). These results suggest that backyard chicken farms are most at risk for transmission of HPAIV compared to farms of the other poultry production types. Since visits of farm-to-farm were high, backyard farms could also a potential source for HPAIV transmission to commercial poultry farms.

Susceptibility of wild passerines to subtype H5N1 highly pathogenic avian influenza viruses

Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype have spread throughout many areas of Asia, Europe and Africa, and numerous cases of HPAI outbreaks in domestic and wild birds have been reported. Although recent studies suggest that the dissemination of H5N1 viruses is closely linked to the migration of wild birds, information on the potential for viral infection in species other than poultry and waterfowl is relatively limited. To investigate the susceptibility of terrestrial wild birds to infection with H5N1 HPAI viruses, common reed buntings (Emberiza schoeniclus), pale thrushes (Turdus pallidus) and brown-eared bulbuls (Hypsipetes amaurotis) were infected with A/mountain hawk-eagle/Kumamoto/1/07(H5N1) and A/whooper swan/Aomori/1/08(H5N1). The results showed that common reed buntings and brown-eared bulbuls were severely affected by both virus strains (100% mortality). While pale thrushes did not exhibit any clinical signs, seroconversion was confirmed. In common reed buntings, intraspecies-transmission of A/whooper swan/Aomori/1/08 to contact birds was also confirmed. The findings show that three passerine species; common reed buntings, brown-eared bulbuls and pale thrushes are susceptible to infection by H5N1 HPAI viruses, which emphasizes that continued surveillance of species other than waterfowl is crucial for effective monitoring of H5N1 HPAI virus outbreaks.

Exposure to a Low Pathogenic A/H7N2 Virus in Chickens Protects against Highly Pathogenic A/H7N1 Virus but Not against Subsequent Infection with A/H5N1

Recent evidences have demonstrated that the presence of low pathogenic avian influenza viruses (LPAIV) may play an important role in host ecology and transmission of avian influenza viruses (AIV). While some authors have clearly demonstrated that LPAIV can mutate to render highly pathogenic avian influenza viruses (HPAIV), others have shown that their presence could provide the host with enough immunological memory to resist re-infections with HPAIV. In order to experimentally study the role of pre-existing host immunity, chickens previously infected with H7N2 LPAIV were subsequently challenged with H7N1 HPAIV. Pre-infection of chickens with H7N2 LAPIV conferred protection against the lethal challenge with H7N1 HPAIV, dramatically reducing the viral shedding, the clinical signs and the pathological outcome. Correlating with the protection afforded, sera from chickens primed with H7N2 LPAIV reacted with the H7-AIV subtype in hemagglutination inhibition assay and specifically with the N2-neuraminidase antigen. Conversely, subsequent exposure to H5N1 HPAIV resulted in a two days-delay on the onset of disease but all chickens died by 7 days post-challenge. Lack of protection correlated with the absence of H5-hemagglutining inhibitory antibodies prior to H5N1 HPAIV challenge. Our data suggest that in naturally occurring outbreaks of HPAIV, birds with pre-existing immunity to LPAIV could survive lethal infections with HA-homologous HPAIV but not subsequent re-infections with HA-heterologous HPAIV. These results could be useful to better understand the dynamics of AIV in chickens and might help in future vaccine formulations.

Highly pathogenic avian influenza virus among wild birds in Mongolia.

Mongolia combines a near absence of domestic poultry, with an abundance of migratory waterbirds, to create an ideal location to study the epidemiology of highly pathogenic avian influenza virus (HPAIV) in a purely wild bird system. Here we present the findings of active and passive surveillance for HPAIV subtype H5N1 in Mongolia from 2005–2011, together with the results of five outbreak investigations. In total eight HPAIV outbreaks were confirmed in Mongolia during this period. Of these, one was detected during active surveillance employed by this project, three by active surveillance performed by Mongolian government agencies, and four through passive surveillance. A further three outbreaks were recorded in the neighbouring Tyva Republic of Russia on a lake that bisects the international border. No HPAIV was isolated (cultured) from 7,855 environmental fecal samples (primarily from ducks), or from 2,765 live, clinically healthy birds captured during active surveillance (primarily shelducks, geese and swans), while four HPAIVs were isolated from 141 clinically ill or dead birds located through active surveillance. Two low pathogenic avian influenza viruses (LPAIV) were cultured from ill or dead birds during active surveillance, while environmental feces and live healthy birds yielded 56 and 1 LPAIV respectively. All Mongolian outbreaks occurred in 2005 and 2006 (clade 2.2), or 2009 and 2010 (clade 2.3.2.1); all years in which spring HPAIV outbreaks were reported in Tibet and/or Qinghai provinces in China. The occurrence of outbreaks in areas deficient in domestic poultry is strong evidence that wild birds can carry HPAIV over at least moderate distances. However, failure to detect further outbreaks of clade 2.2 after June 2006, and clade 2.3.2.1 after June 2010 suggests that wild birds migrating to and from Mongolia may not be competent as indefinite reservoirs of HPAIV, or that HPAIV did not reach susceptible populations during our study.

Modeling Exotic Highly Pathogenic Avian Influenza Virus Entrance Risk Through Air Passenger Violations

The highly pathogenic avian influenza virus (HPAIV) is able to survive in poultry products and could be carried into a country by air travelers. An assessment model was constructed to estimate the probability of the exotic viable HPAIV entering Taiwan from two neighboring areas through poultry products carried illegally by air passengers at Taiwan's main airports. The entrance risk was evaluated based on HPAIV-related factors (the prevalence and the incubation period of HPAIV; the manufacturing process of poultry products; and the distribution-storage-transportation factor event) and the passenger event. Distribution functions were adopted to simulate the probabilities of each HPAIV factor. The odds of passengers being intercepted with illegal poultry products were estimated by logistic regression. The Monte Carlo simulation established that the risk caused by HPAIV-related factors from area A was lower than area B, whereas the entrance risk by the passenger event from area A was similar to area B. Sensitivity analysis showed that the incubation period of HPAIV and the interception of passenger violations were major determinants. Although the result showed viable HPAIV was unlikely to enter Taiwan through meat illegally carried by air passengers, this low probability could be caused by incomplete animal disease data and modeling uncertainties. Considering the negative socioeconomic impacts of HPAIV outbreaks, strengthening airport quarantine measures is still necessary. This assessment provides a profile of HPAIV entrance risk through air travelers arriving from endemic areas and a feasible direction for quarantine and public health measures.

Clade 2.3.2 Avian Influenza Virus (H5N1), Qinghai Lake Region, China, 2009–2010

To the Editor: In 2005, a large population of wild migratory birds was infected with highly pathogenic avian influenza (HPAI) virus (H5N1) in the Qinghai Lake region of western People’s Republic of China, resulting in the death of ≈10,000 birds (1,2). On the basis of phylogenetic analysis of the hemagglutinin (HA) gene, the virus was classified as clade 2.2 according to the World Health Organization guidelines. Subsequently, viruses from this clade were found in Mongolia, Russia, Europe, and Africa along the migratory flyways of birds (3,4). This unique distribution of the same clade of HPAI virus (H5N1) through different migratory routes indicates that migratory birds might play a global role in virus dissemination (3,4). In 2006, viruses from the same clade were isolated in the Qinghai Lake region (3). Analysis of viral outbreaks along migratory flyways demonstrated a similar outbreak pattern for the past 4 years (2006–2009) (5). During that period, clade 2.2 avian influenza virus (H5N1) was isolated in China, Mongolia, Russia, Germany, Egypt, and Nigeria; all viruses were closely related to the Qinghai Lake virus. Despite the broad distribution of clade 2.2 viruses in migratory flyways, few isolates of clade 2.2 viruses in local domestic poultry were reported, especially in China (6). Outbreaks of these viruses were reported in poultry in Africa (7). The reason these viruses rarely cause outbreaks in poultry is unknown. During May–June 2009 and 2010, several dead migratory birds were found in the Qinghai Lake region. Nine HPAI viruses (H5N1) were isolated in 2009 and 2 were isolated in 2010 from great cormorants (Phalacrocorax carbo), brown-headed gulls (Chroicocephalus brunnicephalus), great black-headed gulls (Ichthyaetus ichthyaetus), great-crested grebes (Podiceps cristatus), and bar-headed geese (Anser indicus) and serotyped as described (3). HA genes from all 11 isolates were subsequently amplified by using reverse transcription–PCR and sequenced. Phylogenetic analysis of HA sequences and an additional HA gene sequence from the 2009 Qinghai Lake subtype H5N1 virus isolate from a great crested grebe (from the National Avian Influenza Virus Reference Laboratory, Harbin, China) (GenBank accession no. {type:entrez-nucleotide,attrs:{text:CY063318,term_id:301015462,term_text:CY063318}}CY063318) showed that HA genes from all 12 viruses clustered as clade 2.3.2 (Figure); none clustered with clade 2.2 viruses. Additionally, the HA cleavage site in the new isolates is PQRERRRKRG, which is identical to that of clade 2.3.2 viruses. In clade 2.2, the cleavage site is PQRERRRKKRG. Figure Bootstrapped (1,000×) maximum likelihood phylogenetic tree of hemagglutinin genes of avian influenza viruses (H5N1), People’s Republic of China, 2009–2010. Viruses isolated from the plateau pika near Qinghai Lake are indicated ... A bootstrap (1,000×) maximum likelihood tree (8) also demonstrated that Qinghai 2009 and 2010 virus isolates are closely related to those isolated in Mongolia and Uvs Nuur Lake in 2009, as reported by Sharshov et al. (5). Qinghai Lake and Uvs Nuur Lake, which are found along the migratory flyway in central Asia, are major lakes for bird migration and breeding. Many birds fly from Qinghai Lake to Uvs Nuur Lake in the spring. If one considers isolation date and bird species infected, viruses isolated in Mongolia and Russia and our isolates were likely transmitted between the 2 lake regions by bird migration. Moreover, HA sequences are closely related to viruses isolated from wild birds in Hong Kong and Japan during 2007–2008, which are the most recent isolates of clade 2.3.2 viruses before isolation of 2009 Qinghai Lake viruses. These results indicate that viruses in the Qinghai Lake region may be transmitted by wild birds along the migratory flyway in eastern Asia. However, there is no evidence that avian influenza virus (H5N1) is transmitted from eastern Asian (inner China or across the Himalayas) to the Qinghai Lake region. The 2009 and 2010 Qinghai Lake viruses are related to various viruses isolated from plateau pikas near Qinghai Lake (9). In 2007, clade 2.2 and clade 2.3.2 viruses were isolated from plateau pikas, but no clade 2.3.2 viruses were found in aquatic birds. Wild birds, pikas, and other animals near Qinghai Lake share the same environment, and viruses may be transmitted across species. However, surveillance data are limited for wild animals near Qinghai Lake. Therefore, further investigations need to be conducted to clarify relationships among birds, animals, and influenza viruses near Qinghai Lake. Our results and those of Sharshov et al. (5) show that in 2009 HPAI virus (H5N1) began infecting birds along the migratory route near Qinghai Lake and changed from clade 2.2 viruses to clade 2.3 viruses. New outbreaks of HPAI viruses (H5N1) along this migratory flyway should be investigated.