Influenza In Poultry Research Articles

Understanding the emergence of highly pathogenic avian influenza A virus H5N1 in pinnipeds: An evolutionary approach

Highly pathogenic influenza A virus (HPIAV) H5N1 within the genetic clade 2.3.4.4b has emerged in wild birds in different regions of the world, leading to the death of >70 million birds. When these strains spread to pinniped species a remarkable mortality has also been observed. A detailed genetic characterization of HPIAV isolated from pinnipeds is essential to understand the potential spread of these viruses to other mammalian species, including humans. To gain insight into these matters a detailed phylogenetic analysis of HPIAV H5N1 2.3.4.4b strains isolated from pinniped species was performed. The results of these studies revealed multiple transmission events from birds to pinnipeds in all world regions. Different evolutionary histories of different genes of HPIAV H5N1 2.3.4.4b strains gave rise to the viruses infecting pinnipeds in different regions of the world. European strains isolated from pinnipeds represent a completely different genetic lineage from strains isolated from South American ones. All strains isolated from pinnipeds bear characteristics of a highly pathogenic form for of avian influenza in poultry. Amino acid substitutions, previously shown to confer an adaptive advantage for infecting mammals, were observed in different genes in all pinniped species studied.

Mass vaccination with reassortment-impaired live H9N2 avian influenza vaccine

Avian influenza poses a severe threat to poultry production and global food security, prompting the development of vaccination programs in numerous countries. Modified live virus (MLV) vaccines, with their potential for mass application, offer a distinct advantage over existing options. However, concerns surrounding reversion, recombination, and unintended transmission have hindered the progress of MLV development for avian influenza in poultry. To address these concerns, we engineered reassortment-impaired, non-transmissible, safe, immunogenic, and protective MLVs through the rearrangement of internal gene segments and additional modifications to the surface gene segments HA and NA. The unique peptide marker aspartic acid-arginine-proline-alanine-valine-isoleucine-alanine-asparragine (DRPAVIAN) was incorporated into HA, while NA was modified to encode the chicken interleukin-18 (ckIL18) gene (MLV-H9N2-IL). In vitro, the MLV-H9N2 and MLV-H9N2-IL candidates demonstrated stability and virus titers comparable to the wild-type H9N2 strain. In chickens, the MLV-H9N2 and MLV-H9N2-IL candidates did not transmit via direct contact. Co-infection studies with wild-type virus confirmed that the altered HA and NA segments exhibited fitness disadvantages and did not reassort. Vaccinated chickens showed no clinical signs upon vaccination, all seroconverted, and the inclusion of ckIL18 in the MLV-H9N2-IL vaccine enhanced neutralizing antibody production. A significant decrease in viral loads post-challenge underscored the protective effect of the MLVs. The MLV-H9N2-IL vaccine, administered via drinking water, proved immunogenic in chickens in a dose-dependent manner, generating protective levels of neutralizing antibodies upon aggressive homologous virus challenge. In summary, this study lays the groundwork for safe MLVs against avian influenza suitable for mass vaccination efforts.

High Prevalence of Highly Pathogenic Avian Influenza: A Virus in Vietnam's Live Bird Markets.

In recent years, Vietnam has suffered multiple epizootics of influenza in poultry. From 10 January 2019 to 26 April 2021, we employed a One Health influenza surveillance approach at live bird markets (LBMs) and swine farms in Northern Vietnam. When the COVID-19 pandemic permitted, each month, field teams collected oral secretion samples from poultry and pigs, animal facility bioaerosol and fecal samples, and animal worker nasal washes at 4 LBMs and 5 swine farms across 5 sites. Initially samples were screened with molecular assays followed by culture in embryonated eggs (poultry swabs) or Madin-Darby canine kidney cells (human or swine swabs). Many of the 3493 samples collected had either molecular or culture evidence for influenza A virus, including 314 (37.5%) of the 837 poultry oropharyngeal swabs, 144 (25.1%) of the 574 bioaerosol samples, 438 (34.9%) of the 1257 poultry fecal swab samples, and 16 (1.9%) of the 828 human nasal washes. Culturing poultry samples yielded 454 influenza A isolates, 83 of which were H5, and 70 (84.3%) of these were highly pathogenic. Additionally, a positive human sample had a H9N2 avian-like PB1 gene. In contrast, the prevalence of influenza A in the swine farms was much lower with only 6 (0.4%) of the 1700 total swine farm samples studied, having molecular evidence for influenza A virus. This study suggests that Vietnam's LBMs continue to harbor high prevalences of avian influenza A viruses, including many highly pathogenic H5N6 strains, which will continue to threaten poultry and humans.

Annual report on surveillance for avian influenza in poultry and wild birds in Member States of the European Union in 2022.

All European Union (EU) Member States (MSs) are required to implement surveillance for avian influenza (AI) in poultry and wild birds and (i) to notify the outbreaks, when relevant and (ii) to report the results to the responsible authority. In addition, Iceland, Norway, Switzerland and the United Kingdom (Northern Ireland) also implement ongoing surveillance programmes to monitor occurrences of avian influenza viruses (AIVs) in poultry and wild birds. EFSA received a mandate from the European Commission to collate, validate, analyse and summarise the data resulting from these AI surveillance programmes in an annual report. The present report summarises the results of the surveillance activities carried out in MSs, Iceland, Norway, Switzerland and the United Kingdom (Northern Ireland) in 2022. Overall, the 31 reporting countries (RCs) sampled 22,171 poultry establishments (PEs) during the 2022 surveillance activity: 18,490 PEs were sampled for serological testing and 3775 were sampled for virological testing. Some PEs were therefore sampled for both type of analytical methods. Out of the 18,490 PEs sampled for serological testing, 15 (0.08%) were seropositive for influenza A(H5) viruses. Out of the 3775 PEs sampled for virological testing, 74 PEs (1.96%) were positive to the virological assay for influenza A(H5) viruses. Seropositive PEs were found in four RCs (Belgium, Poland, Spain and Sweden) and as in previous years, the highest percentages of seropositive PEs were found in PEs raising breeding geese and waterfowl game birds. Out of these 15 seropositive PEs, 3 also tested positive by polymerase chain reaction (PCR) for influenza A (H5) viruses - 2 for highly pathogenic avian influenza virus (HPAIV) and 1 low pathogenic avian influenza (LPAI) (H5N3). In relation to the virological surveys, 10 RCs (32%) out of the 31 reported the detection of A (H5) viruses in 74 PEs, covering 12 different poultry categories. More specifically, 54 reported HPAIV A(H5N1), 17 HPAIV (H5N8), 2 AIV (H5N1) with unknown virus pathogenicity and 1 low pathogenic avian influenza (LPAI) (H5N3). Additionally, six PEs tested positive for undefined AIVs in three RCs. A total of 32,143 wild birds were sampled, with 4163 (12.95%) wild birds testing positive for HPAIVs by PCR, from 25 RCs. In contrast to previous years, out of the 4163 wild birds testing positive for HPAIv, subtype A(H5N1) virus was the main influenza A virus subtype identified among the wild bird testing positive for HPAIVs (3942; 95%). In addition, RCs also reported 984 wild birds testing positive for low pathogenic avian influenza (LPAI). Out of those, for 660 (67%) it was ascertained that the subtype was non-A(H5/H7); 260 (26%) wild birds tested positive for LPAIv of A(H5 or H7) subtypes and the remaining 64 (7%) LPAI viruses were belonging to other H-subtypes.

Evaluating Effects of AIV Infection Status on Ducks Using a Flow Cytometry-Based Differential Blood Count.

Ducks have recently received a lot of attention from the research community due to their importance as natural reservoirs of avian influenza virus (AIV). Still, there is a lack of tools to efficiently determine the immune status of ducks. The purpose of this work was to develop an automated differential blood count for the mallard duck (Anas platyrhynchos), to assess reference values of white blood cell (WBC) counts in this species, and to apply the protocol in an AIV field study. We established a flow cytometry-based duck WBC differential based on a no-lyse no-wash single-step one-tube technique, applying a combination of newly generated monoclonal antibodies with available duck-specific as well as cross-reacting chicken markers. The blood cell count enables quantification of mallard thrombocytes, granulocytes, monocytes, B cells, CD4+ T cells (T helper) and CD8+ cytotoxic T cells. The technique is reproducible, accurate, and much faster than traditional evaluations of blood smears. Stabilization of blood samples enables analysis up to 1 week after sampling, thus allowing for evaluation of blood samples collected in the field. We used the new technique to investigate a possible influence of sex, age, and AIV infection status on WBC counts in wild mallards. We show that age has an effect on the WBC counts in mallards, as does sex in juvenile mallards. Interestingly, males naturally infected with low pathogenic AIV showed a reduction of lymphocytes (lymphocytopenia) and thrombocytes (thrombocytopenia), which are both common in influenza A infection in humans. IMPORTANCE Outbreaks of avian influenza in poultry and humans are a global public health concern. Aquatic birds are the primary natural reservoir of avian influenza viruses (AIVs), and strikingly, AIVs mainly cause asymptomatic or mild infection in these species. Hence, immunological studies in aquatic birds are important for investigating variation in disease outcome of different hosts to AIV and may aid in early recognition and a better understanding of zoonotic events. Unfortunately, immunological studies in these species were so far hampered by the lack of diagnostic tools. Here, we present a technique that enables high-throughput white blood cell (WBC) analysis in the mallard and report changes in WBC counts in wild mallards naturally infected with AIV. Our protocol permits large-scale immune status monitoring in a widespread wild and domesticated duck species and provides a tool to further investigate the immune response in an important reservoir host of zoonotic viruses.

Highly Pathogenic Avian Influenza H5N8 Outbreak in Backyard Chickens in Serbia

In winter 2016/2017, the highly pathogenic avian influenza virus H5N8 was detected in backyard poultry in Serbia for the first time. The second HPAI outbreak case in backyard poultry was reported in 2022, caused by subtype H5N1. This is the first study that documents the laboratory identification and pathology associated with highly pathogenic avian influenza in poultry in Serbia during the first and second introduction waves. In both cases, the diagnosis was based on real-time reverse transcriptase PCR. The most common observed lesions included subepicardial hemorrhages, congestion and hemorrhages in the lungs, and petechial hemorrhages in coelomic and epicardial adipose tissue. Histologically, the observed lesions were mostly nonpurulent encephalitis accompanied by encephalomalacia, multifocal necrosis in the spleen, pancreas, and kidneys, pulmonary congestion, and myocardial and pulmonary hemorrhages. In H5N8-infected chickens, immunohistochemical examination revealed strong positive IHC staining in the brain and lungs. Following these outbreaks, strict control measures were implemented on farms and backyard holdings to prevent the occurrence and spread of the disease. Extensive surveillance of birds for avian influenza virus did not detect any additional cases in poultry. These outbreaks highlight the importance of a rapid detection and response system in order to quickly suppress outbreaks.

The Epidemiology of Avian Influenza and its Significance for Public Health

One of the current greatest risks to public health is avian influenza (AI), which originated from the animal reservoir. Therefore the objective of this seminar is to describe the available of epidemiological aspects of AI and to concisely mention public health importance of avian influenza. Although influenza infections in poultry, mainly chickens and turkeys, might go undetected, they frequently lead to production losses and a variety of clinical illnesses ranging in severity from moderate to severe in flocks that are affected. Both the fecal-oral pathway and aerosols can be used to disseminate avian influenza viruse.The majority of LPAI virus transmission occurs through the respiratory route, despite the fact that fecal shedding might occasionally be considerable. Poultry influenza Viruses are categorized as Highly Pathogenic Avian Influenza (HPAI) or Low Pathogenicity Avian Influenza (LPAI) based on how sick and potentially fatal they can make chickens in a laboratory environment. Serious respiratory symptoms, pneumonia, and even mortality, can be brought on by HPAI viruses. By isolating the virus in eggs or cell cultures, or by checking for viral protein, viral nucleic acid, or AI virus antibodies, it is feasible to determine whether someone has been exposed to the AI virus or has become infected with it.Both adamantine and neuraminidase inhibitors, which both block a viral protein, are examples of antiviral drugs that used to prevent and cure influenza. Therefore, for disasters, there should be enabling legislation and an emergency management system (i.e., HPAI incursion) and should be made prompt medical attention and yearly vaccinations.

Құстар миграциясының құс тұмауы вирусының Қазақстанда таралуына әсері

Avian influenza is one of the most dangerous known diseases of poultry. Due to the complete extinction of the entire poultry population, the spread of influenza in poultry farms usually leads to significant economic losses throughout the world. Migratory birds, especially waterfowl, are a natural reservoir of the avian influenza virus. During migration, they exchange with different strains of the virus, causing antigenic drift and antigenic migration, which leads to the emergence of highly pathogenic avian influenza viruses. Their seasonal migration plays an important role in the formation of the genetic structure of the viral population. This requires constant observation of the evolution of the avian influenza virus and regular monitoring in various places to prevent new outbreaks of epizootic and epidemic diseases.

Hand function following accidental automatic animal syringe injector injuries

Accidental self-injection injury is a common occurrence among veterinary and farm workers handling automatic syringe injectors. Most of the time, these injuries are asymptomatic or cause self-resolving mild symptoms, but these injuries may lead to significant morbidity. The aim of the study was to evaluate hand function after inadvertent injection of a poultry influenza or cholera vaccine in patients admitted to our department with infection. We retrospectively gathered data from admission to last follow-up. Functional assessment and physical exam of the hand were done at each stage by either an orthopedic resident or a fellowship-trained hand surgeon. The exam included evaluation of sensation using monofilament, joint range of motion using a goniometer, and a Quick DASH questionnaire. The study included 21 patients, all men, with a mean age of 33.4 years (range 23–44). Of the 21 patients only eight had attended all follow-ups. All patients had injury to the non-dominant hand. Seventeen of 21 of the cases had finger injuries, out of which 11 involved the thumb. The mean hospitalization time was 3.75 days (1–10). Of the 21 patients, seven underwent surgery to drain a collection during hospitalization. Seven out of eight patients had lowest disability scores on Quick Dash questionnaire. Three out of eight patients lost superficial sensation at the tip of the finger. The largest loss of range of motion was found in the distal interphalangeal joint in the finger or interphalangeal joint in the thumb, especially following surgical drainage. Of the eight patients presenting for follow-up, most had returned to the same job. Hand function was normal, as expressed in a DASH questionnaire. Sensory examination demonstrated that the sensation was almost unaffected over the injured finger. Range of motion of the joint closest to the injection site was usually the most impaired. Patients who underwent surgical drainage had a reduced range of motion.

Annual report on surveillance for avian influenza in poultry and wild birds in Member States of the European Union in 2021.

European Union (EU) Member States (MSs) are required to carry out surveillance for avian influenza (AI) in poultry and wild birds and notify the results to the responsible authority. In addition, Iceland, Norway, Switzerland and the United Kingdom (Northern Ireland) also implement ongoing surveillance programmes to monitor incursions of avian influenza viruses (AIVs) in poultry and wild birds. EFSA received a mandate from the European Commission to collate, validate, analyse and summarise the data resulting from these AI surveillance programmes in an annual report. The present report summarises the results of the surveillance activities carried out in MSs and the aforementioned countries in 2021. Overall, 24,290 poultry establishments (PEs) were sampled, of which 27 were seropositive for influenza A(H5) and 4 for A(H7) viruses. Seropositive PEs were found in 10 MSs and, as per previous years, the highest percentages of seropositive PEs were found in establishments raising waterfowl game birds and breeding geese. Out of these 31 seropositive PEs, 3 tested positive by polymerase chain reaction (PCR) for influenza A(H5) viruses: 1 for highly pathogenic avian influenza virus (HPAIV), 1 for low pathogenic avian influenza virus (LPAIV) and 1 with unknown virus pathogenicity. In addition, 16 countries reported PCR test results from 1,858 PEs which did not correspond to the follow‐up testing of a positive serology event (e.g. in some PEs, PCR tests were used for screening). Sixty‐five of these PEs in 10 MSs were found positive for AIVs. Apart from poultry, 31,382 wild birds were sampled, with 2,314 wild birds testing positive for HPAIVs by PCR. Twenty‐two countries reported HPAIV‐positive wild birds and most positive samples were identified as highly pathogenic avian influenza (HPAI) A(H5N8) virus. In addition, 328 wild birds tested positive for LPAIVs of the A(H5/H7) subtypes and 362 wild birds tested positive for non‐A(H5/H7) subtype AIVs.

Annual Report on surveillance for avian influenza in poultry and wild birds in Member States of the European Union in 2020.

In 2020, Council Directive 2005/94/EC required EU Member States (MSs) to carry out surveillance for avian influenza (AI) in poultry and wild birds and notify the results to the responsible authority. Based on this, MSs, Iceland, Norway, Switzerland and the United Kingdom implemented ongoing surveillance programmes to monitor incursions of AI viruses in poultry and wild birds. EFSA received a mandate from the European Commission to collate, validate, analyse and summarise the data resulting from the avian influenza surveillance programmes in an annual report. This is the second such report produced using data directly submitted to EFSA by MSs. This report summarises the results of the surveillance activities carried out in poultry and wild birds in 2020. Overall, 24,768 poultry establishments (PEs) were sampled, of which 46 were seropositive for H5 virus strains and seven for H7 strains. Seropositive PEs were found in nine MSs (Belgium, Denmark, Finland, France, Italy, the Netherlands, Poland, Spain and Sweden) and the United Kingdom. As per previous years, the highest percentages of seropositive PEs were found in establishments raising waterfowl game birds and breeding geese. Out of the 53 PEs with positive serological tests for H5/H7, seven tested positive in polymerase chain reaction (PCR) or virology for H5/H7 virus strains: six for Low Pathogenic Avian Influenza (LPAI) and one for Highly Pathogenic Avian Influenza (HPAI). In addition, 13 countries also reported PCR results from 748 PEs which did not correspond to the follow‐up testing of a positive serology event (e.g. in some PEs, PCR tests were used for screening). Twenty‐five of these PEs were found positive for AI viral RNA. These positive PEs were located in Bulgaria, Estonia, Germany, Romania and Slovakia. A total of 18,968 wild birds were sampled, with 878 birds testing positive to HPAI virus. Fourteen countries reported HPAI‐positive wild birds, with all HPAI strains identified as H5. Most positive birds were infected with H5N8, with a smaller number of N1, N3, N5 and unidentified NA subtypes. In addition, there were 317 birds testing positive for LPAI H5 or H7 virus and 429 birds testing positive for non‐H5/H7 AI virus, reported by 31 countries. The surveillance findings for poultry and wild birds for 2020 are discussed in relation to the current knowledge of the epidemiology of AI in Europe, in particular the H5N8 epidemic which has been identified late 2020.

Detection of avian influenza-infected chickens based on a chicken sound convolutional neural network

The modern poultry industry is large-scale and breeding-intensive, making the spread of disease in poultry easier, faster and more harmful. Avian influenza (AI) is the most important disease in poultry, and prevention and detection of avian influenza in poultry is a focus of scientific research and the poultry industry. In this paper, a new sound recognition method, the chicken sound convolutional neural network (CSCNN), is proposed for detection of chickens with avian influenza. According to the spectral differences in environmental noise, chicken behaviour noise and chicken sound, a method was designed to extract the chicken sound from complex sound data. Four features of the chicken sounds were calculated and combined into feature maps, including Logfbank, Mel Frequency Cepstrum Coefficient (MFCC), MFCC Delta and MFCC Delta-Delta. Finally, the sounds of healthy chickens and chickens with avian influenza were recognized using CSCNN. In the experiment, the recognition accuracies of CSCNN via spectrogram (CSCNN-S) were 93.01%, 95.05%, and 97.43% on the 2nd, 4th, and 6th day after injection with the H9N2 virus, and the recognition accuracies of CSCNN with feature mapping (CSCNN-F) were 89.79%, 93.56%, and 95.84%, respectively. The experimental results show that the method proposed in this paper can be used to quickly and effectively detect avian influenza-infected chickens via chicken sound.

The Presence of Poultry Influenza Strains in Two Live Bird Markets near the East-West Boundary of Vietnam.

The spread of avian influenza virus among Asian countries is becoming a concern after influenza epidemics in recent years. This study is aimed at identifying the subtypes of avian influenza viruses collected from healthy chickens and ducks at two live bird markets in a border province of Vietnam and the Lao People Democratic Republic. Cloacal and tracheal swab samples from 100 chickens and 101 ducks were collected in May 2017. All samples were screened to detect avian influenza virus by real-time reverse transcriptase PCR. Samples that are avian influenza virus-positive were isolated in embryonated chicken eggs, and the subtypes were identified by RT-PCR with the specific primers. The samples positive for influenza virus H5 were sequenced to identify HA and NA genes. The prevalence of avian influenza virus (AIV) among chicken and duck samples was 27.5% (55/200) and 24.8% (50/202), respectively. AIV subtypes identified among 17 samples positive with the hemagglutination test include H3N6, H6N6, and H9N2. Of these 17 samples, 7 duck samples were found to be H6N6, 4 duck samples were infected with both subtypes of H3N6 and H6N6, and two chicken samples were recorded as H9N2. A positive chicken sample with A/H5 contains 99% similarity nucleotide with H5N6 reference strain. Results suggested that while the presence of low pathogenic avian influenza virus is predominant, potential risks of the appearance of high pathogen avian influenza virus in the east-west boundary in Vietnam should be concerned and studied further. Furthermore, prevention activities are needed to reduce such biosecurity threats in Vietnam and other Asian countries.

Development of a Multiplex RT-qPCR for the Detection of Different Clades of Avian Influenza in Poultry.

Since the initial detection of H5N1, a highly pathogenic avian influenza (HPAI) virus, in 1996 in China, numerous HPAI H5 lineages have been classified, and they continue to pose a threat to animal and human health. In this study, we developed a novel primer/probe set that can be employed to simultaneously detect pan-H5 HPAI and two clades, 2.3.2.1 and 2.3.4.4, of H5Nx viruses using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The sensitivity and specificity of these primer sets and probes were confirmed with a number of different subtypes of influenza virus and the H5-HA gene plasmid DNA. In particular, the multiplex RT-qPCR assay was successfully applied to the simultaneous detection of H5 HPAI and different virus clades in clinical field samples from a poultry farm. Therefore, this multiplex assay and a novel detection primer set and probes will be useful for the laboratory diagnosis and epidemiological field studies of different circulating H5 HPAI virus clades in poultry and migratory wild birds.

Annual Report on surveillance for avian influenza in poultry and wild birds in Member States of the European Union in 2018.

Avian influenza (AI) is a viral infectious disease that affects all species of domestic and wild birds. The viruses causing this disease can be of high (HPAI) or low (LPAI) pathogenicity and represent a continuous threat to poultry in Europe. Council Directive 2005/94/EC requests EU Member States (MSs) to carry out surveillance in poultry and wild birds and notify the results to the responsible authority. Therefore, MSs and Switzerland have implemented surveillance programmes to yearly monitor incursions of AI viruses in poultry and wild birds. EFSA received a mandate from the European Commission, to collate, validate, analyse and summarise in an annual report the data resulting from the avian influenza surveillance programmes. This is the first report produced under this mandate summarising the results of the surveillance activities carried out in poultry and wild birds in 2018. Overall 18,596 poultry establishments were sampled, of which 43 were seropositive for H5 AI and two for H7 AI. Seropositive establishments were found in 11 MSs, with the highest percentage of seropositive establishments being found in waterfowl gamebird, and geese and duck breeding establishments. A total of 9,145 dead/moribund wild birds were sampled, with 163 birds testing positive to HPAI virus H5N6. The infected birds were reported by eight MSs and were mostly found between January and April 2018. In this report, the wild bird species affected with HPAI are described and the strategy of targeted sampling is assessed. The crude odds ratio of HPAI detection as a function of the target species (species belonging to the list of target species versus species not belonging to the target list) is presented. The surveillance findings for poultry and wild birds for 2018 are also discussed in relation to findings from previous years and current knowledge on the epidemiology of AI in Europe.

SPREAD OF LOW PATHOGENIC AVIAN INFLUENZA А/Н9N2 IN THE WORLD AND RUSSIAN FEDERATION. CHALLENGES OF DISEASE ERADICATION

The paper demonstrates data on global and national spread of subtype H9 low pathogenic avian influenza in poultry. Due to no need of the mandatory disease notification to the OIE, published data are used for the analysis of the infection spread. Cases of combined low pathogenic avian influenza clinical manifestation in poultry population are described. Challenges of the infection eradication are addressed. Global spread of А/Н9 virus in poultry, its rapid evolution and zoonotic potential require close attention. Since the disease signs and clinical course are similar to the ones of other avian infectious diseases (Newcastle disease, infectious laryngotracheitis, infectious bronchitis, metapneumovirus infection, mycoplasmosis, infectious coryza, etc.), the routine laboratory diagnosis should include tests for low pathogenic avian influenza thus facilitating the rapid response to the threat. Н9 virus circulation is reflected not only in the physical condition of birds and their productivity, but it also affects the live vaccines’ efficacy during scheduled immunizations through the reduction of their protective effect and increase of post-vaccination reactions. In spite of the availability of specific prevention, the opinions of the researchers divided between its opponents and proponents. On the one hand, the vaccination eliminates the economic loses and the virus escape into the environment, but on the other hand, it can aid to “silent” spread of the virus and its established variation that is typical in case of low strength of the poultry population immunity.

Efficacy of a plant-produced virus-like particle vaccine in chickens challenged with Influenza A H6N2 virus.

SummaryThe efficacy, safety, speed, scalability and cost‐effectiveness of producing hemagglutinin‐based virus‐like particle (VLP) vaccines in plants are well‐established for human influenza, but untested for the massive poultry influenza vaccine market that remains dominated by traditional egg‐grown oil‐emulsion whole inactivated virus vaccines. For optimal efficacy, a vaccine should be closely antigenically matched to the field strain, requiring that influenza A vaccines be updated regularly. In this study, an H6 subtype VLP transiently expressed in Nicotiana benthamiana was formulated into a vaccine and evaluated for efficacy in chickens against challenge with a heterologous H6N2 virus. A single dose of the plant‐produced H6 VLP vaccine elicited an immune response comparable to two doses of a commercial inactivated H6N2 vaccine, with mean hemagglutination inhibition titres of 9.3 log2 and 8.8 log2, respectively. Compared to the non‐vaccinated control, the H6 VLP vaccine significantly reduced the proportion of shedders and the magnitude of viral shedding by >100‐fold in the oropharynx and >6‐fold in the cloaca, and shortened oropharyngeal viral shedding by at least a week. Despite its potency, the cost of the antigenic mismatch between the inactivated H6N2 vaccine and challenge strain was evident not only in this vaccine's failure to reduce viral shedding compared to the non‐vaccinated group, but its apparent exacerbation of oropharyngeal viral shedding until 21 days post‐challenge. We estimate that a kilogram of plant leaf material can produce H6 VLP vaccines sufficient for between 5000 and 30 000 chickens, depending on the effective dose and whether one or two immunizations are administered.

Effective mosaic-based nanovaccines against avian influenza in poultry

Avian influenza virus (AIV) is an extraordinarily diverse pathogen that causes significant morbidity in domesticated poultry populations and threatens human life with looming pandemic potential. Controlling avian influenza in susceptible populations requires highly effective, economical and broadly reactive vaccines. Several AIV vaccines have proven insufficient despite their wide use, and better technologies are needed to improve their immunogenicity and broaden effectiveness. Previously, we developed a “mosaic” H5 subtype hemagglutinin (HA) AIV vaccine and demonstrated its broad protection against diverse highly pathogenic H5N1 and seasonal H1N1 virus strains in mouse and non-human primate models. There is a significant interest in developing effective and safe vaccines against AIV that cannot contribute to the emergence of new strains of the virus once circulating in poultry. Here, we report on the development of an H5 mosaic (H5M) vaccine antigen formulated with polyanhydride nanoparticles (PAN) that provide sustained release of encapsulated antigens. H5M vaccine constructs were immunogenic whether delivered by the modified virus Ankara (MVA) strain or encapsulated within PAN. Both humoral and cellular immune responses were generated in both specific-pathogen free (SPF) and commercial chicks. Importantly, chicks vaccinated by H5M constructs were protected in terms of viral shedding from divergent challenge with a low pathogenicity avian influenza (LPAI) strain at 8 weeks post-vaccination. In addition, protective levels of humoral immunity were generated against highly pathogenic avian influenza (HPAI) of the similar H5N1 and genetically dissimilar H5N2 viruses. Overall, the developed platform technologies (MVA vector and PAN encapsulation) were safe and provided high levels of sustained protection against AIV in chickens. Such approaches could be used to design more efficacious vaccines against other important poultry infections.

Heterosubtypic protection against avian influenza virus by live attenuated and chimeric norovirus P-particle-M2e vaccines in chickens

Avian influenza in poultry continues to be a great concern worldwide, and the currently licensed inactivated influenza vaccines are not effective against the novel strains of influenza virus that continue to emerge in the field. This warrants the development of more broadly protective influenza vaccines or vaccination regimens. Live attenuated influenza vaccines (LAIVs) and subunit vaccines derived from viral peptides, such as the highly conserved ectodomain of influenza virus matrix protein 2 (M2e), can offer a more broadly reactive immune response. In chickens, we previously showed that a chimeric norovirus P particle containing M2e (M2eP) could provide partial but broad immunity, when administered as a standalone vaccine, and also enhanced the protective efficacy of inactivated vaccine when used in a combination regimen. We also demonstrated that a naturally-selected NS1-truncated H7N3 LAIV (pc4-LAIV) was highly efficacious against antigenically distant heterologous H7N2 low pathogenicity avian influenza virus challenge, especially when used as the priming vaccine in a prime-boost vaccination regimen. In this study, we investigated the cross-subtype protective efficacy of pc4-LAIV in conjunction with M2eP using single vaccination, combined treatment, and prime-boost approaches. Chickens vaccinated with pc4-LAIV showed significant reduction of tracheal shedding of a low pathogenicity H5N2 challenge virus. This cross-subtype protective efficacy was further enhanced, during the initial stages of challenge virus replication, in chickens that received a vaccination regimen consisting of priming with pc4-LAIV at 1 day of age and boosting with M2eP. Further, H5N2-specific serum IgG and pc4-LAIV-specific hemagglutination-inhibition antibody titers were enhanced in LAIV-primed and M2eP boost-vaccinated chickens. Taken together, our data point to the need of further investigation into the benefits of combined and prime-boost vaccination schemes utilizing LAIV and epitope-based vaccines, to develop more broadly protective vaccination regimens.

Evidence of exposure and human seroconversion during an outbreak of avian influenza A(H5N1) among poultry in Cameroon

ABSTRACT From May 2016 to March 2017, 22 poultry outbreaks of avian influenza A(H5N1) were reported in Cameroon, mainly in poultry farms and live bird markets. No human cases were reported. In this study, we sought to describe the 2016 A(H5N1) outbreak strain and to investigate the risk of infection in exposed individuals. We find that highly pathogenic influenza subtype A(H5N1), clade 2.3.2.1c from Cameroon is closely related phylogenetically and antigenically to strains isolated in central and western Africa at the time. No molecular markers of increased human transmissibility were noted; however, seroconversion was detected in two poultry workers (1.5% of total screened). Therefore, the continued outbreaks of avian influenza in poultry and the risk of zoonotic human infection highlight the crucial need for continued and vigilant influenza surveillance and research in Africa, especially in areas of high poultry trade, such as Cameroon.