Commercial Poultry Flocks Research Articles

Virulence-associated genes in faecal and clinical Escherichia coli isolates cultured from broiler chickens in Australia.

A healthy chicken's intestinal flora harbours a rich reservoir of Escherichia coli as part of the commensal microbiota. However, some strains, known as avian pathogenic E. coli (APEC), carry specific virulence genes (VGs) that enable them to invade and cause extraintestinal infections such as avian colibacillosis. Although several VG combinations have been identified, the pathogenic mechanisms associated with APEC are ill-defined. The current study screened a subset of 88 E. coli isolates selected from 237 pre-existing isolates obtained from commercial poultry flocks in Australia. The 88 isolates were selected based on their enterobacterial repetitive intergenic consensus (ERIC) and antimicrobial resistance (AMR) profiles and included 29 E. coli isolates cultured from chickens with colibacillosis (referred to as clinical E. coli or CEC) and 59 faecal E. coli (FEC) isolates cultured from clinically healthy chickens. The isolates were screened for the presence of 35 previously reported VGs. Of these, 34 were identified, with iucA not being detected. VGs focG, hlyA and sfa/foc were only detected in FEC isolates. Eight VGs had a prevalence of 90% or above in the CEC isolates. Specifically, astA (100%); feoB (96.6%); iutA, iss, ompT, iroN and hlyF (all 93.1%); and vat (89.7%). The prevalence of these were significantly lower in FEC isolates (astA 79.7%, feoB 77.9%, iutA 52.5%, iss 45.8%, ompT 50.9%, iroN 37.3%, hlyF 50.9% and vat 42.4%). The odds ratios that each of these eight VGs were more likely to be associated with CEC than FEC ranged from 7.8 to 21.9. These eight VGs may be used to better define APEC and diagnostically detect APEC in Australia. Further investigations are needed to identify the roles of these VGs in pathogenicity.

Assessment of Different Newcastle Disease Virus Antigens and Inactivators of Binary Ethylene Amine and Formalin for the Hemagglutination Inhibition Assay.

Newcastle disease is a severe viral threat to the global poultry industry due to its high prevalence and rapid transmission. Evaluating vaccination timing and effectiveness is crucial, often accomplished through the hemagglutination inhibition (HI) assay. This test relies on the virus's agglutination ability in certain animals, utilizing various inactivated antigens. Our study aimed to assess multiple Newcastle viral antigens ( LaSota, clone, thermo-resistant strain, B1, and V4 ) inactivated by binary ethylene amine (BEA) and formalin, seeking the best antigen and inactivator for the HI assay. We prepared the different ND antigens include; LaSota, Clone, thermo resistant, B1, V4 and the mixture of the antigens then inactivated them using BEA and formalin. The hemagglutination (HA) assay determined mean titers, comparing BEA and formalin inactivation. These antigens were also subjected to the HI test using 112 serum samples from different commercial poultry flocks to assess their performance. BEA-inactivated antigens exhibited significantly higher mean titers in the HA assay than formalin-inactivated antigens. In the evaluation of different antigens in the HI test, the mean titer of antigen B1 followed by clone and LaSota displayed a higher mean titer than others. In conclusion, this study recommends using Hitchner pathotype antigens, specifically the B1 vaccine, for Newcastle disease HI testing. BEA is the preferred inactivator, preserving antigen structure particularly the structure of hemagglutinin antigen while minimizing risks. These findings can enhance serological testing accuracy, contributing to more effective disease control and prevention in the poultry industry.

Virulence evaluation of Israeli Marek's disease virus isolates from commercial poultry using their meq gene sequence.

Fifty-sevenGallid alphaherpesvirus 2 (GaHV-2) isolates, collected during a 30-year period (1990-2019)from commercial poultry flocks affected by Marek's disease (MD), were molecularly characterised. The GaHV-2 meq gene was amplified and sequenced to evaluate the virus virulence, based on the number of PPPPs within the proline-rich repeats (PRRs) of its transactivation domain. The present illustration of virus virulence evaluation on a large scale of field virus isolates by molecular analysisexemplifies the practical benefit and usefulness of the molecular marker in commercial GaVH-2 isolates. The alternative assay of GaVH-2 virulence pathotyping is the classical Gold Standard ADOL method, which is difficult and impossible to employ on a large scale using the Specific Pathogen Free (SPF) chicks of the ADOL strainskept in isolators for two months. The phylogenetic analysis performed in the present study showed that the meq gene amino acid sequences of the 57 Israeli strains divide into 16 phylogenetic branches. The virulence evaluation was performed in comparison with 36 GaHV-2 prototype strains, previously characterised by the in vivo Gold Standard ADOL assay. The results obtained revealed that the GaHV-2 strains circulating in Israel have evolved into a higher virulence potential during the years, as the four-proline stretches number in the meq gene decreased over the investigated period, typically of very virulent virus prototypes. The present study supports the meq gene molecular markers for the assessment of field GaVH-2 strains virulence.

Molecular detection and sequence analysis of envelope glycoproteins (gC, gD) and tegument protein (UL47) of infectious laryngotracheitis virus isolated from commercial poultry flocks in Tamil Nadu, India

Molecular detection and sequence analysis of envelope glycoproteins (gC, gD) and tegument protein (UL47) of infectious laryngotracheitis virus isolated from commercial poultry flocks in Tamil Nadu, India

Pathology and molecular detection of influenza A subtype H9N2 virus in commercial poultry in Nigeria, 2024.

After the first Avian Influenza H5N1 outbreak in Nigerian poultry in 2006, subsequent waves of outbreaks occurred, causing substantial losses. Despite effective control measures by 2008, a resurgence in 2015 led to further losses and required depopulation efforts. The aim of this study was to do pathology and molecular detection of influenza A subtype H9N2 virus in commercial poultry in Nigeria during 2024. In February 2024, a poultry farmer reported high mortality in his mixed commercial poultry flock in Ibadan, Nigeria, submitting carcasses to the University of Ibadan's V.T.H. and the FAO Regional Laboratory at National Veterinary Research Institute (NVRI), Vom. Necropsy of nine Isa Brown layers and three Abor Acre broilers revealed cyanosis of comb and wattles, generalized petechial and ecchymotic hemorrhages including shank hemorrhages with sinusitis, pneumonia, and severe greenish fecal pasting also observed. At histopathology, denudation of the tracheal epithelia and parabronchial epithelial necrosis, obliteration, with airsac edema and emphysema were observed. At NVRI, qPCR detected an Influenza A H9N2 virus in several pooled organ samples of layers, and broilers and eliminating the avian infectious bronchitis and Newcastle disease viruses. This is the first report of an H9N2 outbreak in commercial poultry in Southern Nigeria. The high pathogenicity shown in commercial poultry in this outbreak and the risk of dispersal of infected live poultry in Nigeria as previously seen in H5N1 require stakeholders' intervention.

Risk Factors for Salmonella Detection in Commercial Layer Flocks in Spain.

Trends in Salmonella human infections are assumed to be related to the distribution of the pathogen in the animal reservoir/food products, and cases in humans are most often linked to poultry and poultry products (eggs, meat). Therefore, ongoing Salmonella national control programs (NCPs) in European Union Member States have the objective of monitoring and reducing its prevalence in commercial poultry flocks. Results from NCPs have shown certain factors (housing systems, season of sampling and if sampling is conducted by food business operators (FBOps) or competent authorities (CAs), among others) can influence detection rates, but associations are often not consistent. Here, we analyzed data from the Spanish NCP on 7216 laying hen flocks subjected to 36,193 sampling events over a six-year period to characterize its performance and identify variables influencing detection rates. Overall, 1205 sampling events were positive for Salmonella spp. (any serovar) and 132 for S. Enteritidis-S. Typhimurium/monophasic. Bayesian multivariable models adjusting for multiple covariates concluded that sampling events later in the year, in caged flocks with older animals and conducted by CAs had increased odds of positivity for Salmonella spp., revealing aspects linked with a differential estimation of Salmonella levels in laying hen flocks.

A molecular survey of Chlamydia spp. infection in commercial poultry and detection of Chlamydia pneumoniae in a commercial turkey flock in Iran.

Chlamydiaceae are a group of gram-negative intracellular bacteria which can infect a wide variety of hosts. Some chlamydial agents are capable of crossing the host barrier and though they are potentially a risk to very different species. They also pose a zoonotic risk for human and different chlamydial agents are linked to several medical maladies. In this study, the presence of chlamydial agents in different commercial poultry flocks in Iran was investigated. Swab and tissue samples were collected from 435 birds in 24 different commercial poultry flocks. These samples were examined using a Chlamydiaceae-specific real-time PCR assay targeting 23S rRNA gene. Positive samples then were subjected to intergenic spacer rRNA (IGS) gene and major outer membrane protein gene (ompA) PCRs. Finally, positive PCR products were sequenced and analysed. Only one flock of commercial turkey became positive. Partial DNA sequencing of IGS gene revealed that all positive samples from the infected flock were Chlamydia pneumoniae and were identical to previously studied isolates from koala (LPCoLN) and frog (DC9). Further investigations showed slight dissimilarity in ompA gene of C. pneumoniae from different hosts. The detected turkey isolates were located in a different clade of phylogenetic tree, close to Western barred bandicoot and koala isolates. C. pneumoniae has passed the cross-species barrier in the past and therefor it could potentially be zoonotic. To the best of authors' knowledge, this is the first report of C. pneumoniae infection in commercial turkey.

H5N1 highly pathogenic avian influenza clade 2.3.4.4b in wild and domestic birds: Introductions into the United States and reassortments, December 2021–April 2022

Highly pathogenic avian influenza viruses (HPAIVs) of the A/goose/Guangdong/1/1996 lineage H5 clade 2.3.4.4b continue to have a devastating effect on domestic and wild birds. Full genome sequence analyses using 1369 H5N1 HPAIVs detected in the United States (U.S.) in wild birds, commercial poultry, and backyard flocks from December 2021 to April 2022, showed three phylogenetically distinct H5N1 virus introductions in the U.S. by wild birds. Unreassorted Eurasian genotypes A1 and A2 entered the Northeast Atlantic states, whereas a genetically distinct A3 genotype was detected in Alaska. The A1 genotype spread westward via wild bird migration and reassorted with North American wild bird avian influenza viruses. Reassortments of up to five internal genes generated a total of 21 distinct clusters; of these, six genotypes represented 92% of the HPAIVs examined. By phylodynamic analyses, most detections in domestic birds were shown to be point-source transmissions from wild birds, with limited farm-to-farm spread.

SURVEY ON THE MOST COMMON DISEASES CIRCULATING AMONG PIGEONS IN THE EASTERN PROVINCE, SAUDI ARABIA

Molecular and microbiological approaches were used to investigate the common circulating pathogens in pigeon flocks. Sixty-nine flocks were investigated for viral, bacterial and parasitic infection evidence, including the histories, clinical signs and lesions. We reported that the seroprevalences of Newcastle disease viruses (NDVs), Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) were 50.57%, 7.25% and 8.69%. respectively; no avian influenza virus (AIV)‒specific antibodies were detected. NDVs and pigeon paramyxoviruses (PPVs) were detected in 40.57% and 15.9%, respectively, of the pigeons. Multiple diseases were the predominant finding, with 60.87% of diseased flocks harboring multifactorial infections of viral, bacterial and parasitic pathogens. Concurrent viral and bacterial infections were reported in 15.94%, and viral and parasitic coinfections were found in 20.29%. Mixed infections of NDV with bacterial and parasitic diseases were detected in 21.74%. Bacterial and mycotic pathogens were detected in 22/69 (31.88%) and 5/69 (7.25%), respectively. Salmonella spp. were only detected in 3/69 (4.35%) of diseased flocks. Parasitic diseases were the most prevalent infections, either as a single infection or concurrently with others. Parasites including Cestoda spp., Ascaris spp., Eimeria spp., Trichomonas gallinae and external parasites(Menopon gallinae, Pseudolynchia canariensis) were reported in 12/69 (17.39%), 10/69 (10.49%), 10/69 (10.49%), 18/69 (26.1%) and 3/69 (4.35%), respectively. We concluded that pigeon flocks harbor many pathogens that may threaten the health of animals and humans; they are serious amplifiers and reservoir hosts of disease and, ultimately, may be serious biological hazards to the intensive poultry production sector and community. Pigeon vaccines should be developed. Strict biosecurity measures should be applied to protect pigeons and commercial poultry flocks. Key words: Pigeon, diseases, PCR, isolation and identification, Saudi Arabia

Detection, Quantification and Molecular Characterization of Fowl Adenoviruses Circulating in Ecuadorian Chicken Flocks during 2019-2021.

Fowl adenoviruses are a group of pathogens that cause large economic losses worldwide in the poultry industry, in addition to producing a wide range of diseases, such as IBH, HHS, and enteric and respiratory diseases. The objective of this study was to quantify, identify, and molecularly characterize the types of FAdV circulating in commercial poultry farms (broilers, breeders, and layers) in Ecuador from 2019 to 2021. Molecular characterization was performed by PCR, quantification by qPCR, and subsequent sequencing for each positive sample. The results indicated that the FAdV genotypes circulating in our country are FAdV-2/D2, FAdV-6/E1, FAdV-8a/E2, and FAdV-11/D3; the samples were grouped into different groups that contain sequences that were obtained from countries in Africa, Asia, and America, and that are found in birds at different ages, since early age where can cause different clinical signs, such as diarrhea, ruffled feathers and dwarfism. Therefore, these results indicate that several genotypes of the virus are circulating in commercial poultry flocks, suggesting that biosecurity measures on farms should be improved, in addition to carrying out new or improved vaccination plans.

West Nile Virus-Induced Drop in Egg Production in Commercial Pekin Duck Breeders.

A sudden drop in egg production in commercial poultry flocks can be economically devastating, and rapid identification of the cause often requires a combined effort between the producer, veterinarian, and pathologist. In September 2019, a 35-wk-old commercial Pekin breeder duck flock in Indiana suffered a drop in egg production from 1700 to 1000 eggs daily (41.2% drop). Again, in September 2021, three Pekin breeder duck flocks aged 32, 58, and 62 wk from the same company suffered a similar drop in egg production, with a mild increase in weekly mortality of 1.0% to 2.5%. In 2019 and in 2021, birds from affected flocks were submitted to the Veterinary Diagnostic Laboratory at Michigan State University for postmortem examination. Common gross examination findings included flaccid, shrunken, or atrophied ova (all hens), pododermatitis, airsacculitis, hepatomegaly, splenomegaly, ascites, and pallor of the left ventricle. Histopathologic examination of cerebrum, cerebellum, and brainstem revealed mild lymphocytic perivascular cuffing, vasculitis, and gliosis, suggesting viral encephalitis. In the heart, there was mild multifocal cardiomyocyte necrosis, mineralization, and infiltration by lymphocytes and macrophages. PCR for Newcastle disease virus, avian influenza virus, eastern equine encephalitis virus, and West Nile virus (WNV) was performed. Brain and heart samples were positive for WNV by PCR, and WNV antigen was detected in the cerebellum by immunohistochemistry. This is the first report to associate WNV infection with a drop in egg production in waterfowl, which are known to be important reservoir species for WNV and, as such, are generally asymptomatic.

Essential Oils as an Alternative to Antibiotics to Reduce the Incidence and Severity of Necrotic Enteritis in Broiler Chickens: A Short Review

Due to the removal of antibiotic growth promoters (AGPs) and consumer pressure for antibiotic-free (ABF) or no antibiotics ever (NAE) poultry production, there is a need for sustainable alternatives to prevent disease in commercial poultry operations. Without AGPs, there has been a rise in diseases that were traditionally controlled by subtherapeutic levels of antibiotics in the diet. This has impacted the health of commercial poultry and has been a significant cost to poultry producers. To mitigate this, the industry has started to investigate alternatives to antibiotics to treat these forthcoming health issues, such as necrotic enteritis (NE). NE is an enteric disease caused by an over proliferation of toxigenic Clostridium perfringens (CP) in the gastrointestinal tract. Although CP is a commensal in the avian intestinal tract, dysbiosis caused by inflammation and impaired intestinal integrity facilitates uncontrolled replication of CP. Infectious agents, such as Eimeria maxima, appear to be a predominant predisposing factor that promotes NE. However, non-infectious stressors, including dietary changes, have also been associated with NE to some degree. As a result of increased pressure to restrict the use of antibiotics, there is a need for research evaluating the efficacy of alternatives, such as plant-derived essential oils, as potential tools to mitigate NE in commercial poultry flocks. The aim of this study is to review the effects of essential oils as an alternative to antibiotics to reduce the incidence and severity of necrotic enteritis in broiler chickens.

Evolutionary Trajectories of Avian Avulaviruses and Vaccines Compatibilities in Poultry.

Newcastle disease virus (NDV) causes one of the highly infectious avian diseases in poultry leading to genuine financial misfortunes around the world. Recently, there has been an increasing trend in the number of ND-associated outbreaks in commercial Jordanian poultry flocks indicating a possible complex evolutionary dynamic of NDV infections in the country. To underpin the dynamics of circulating NDV strains and to assess the vaccine-escape potential, a total of 130 samples were collected from different poultry flocks in six Jordanian Governorates during 2019-2021. Twenty positive isolates, based on real-time reverse transcriptase PCR, were used for further genetic characterization and evolutionary analysis. Our results showed that there is a high evolutionary distance between the newly identified NDV strains (genotype VII.1.1) in this study and the commercially used vaccines (genotypes I and II), suggesting that circulating NDV field strains are under constant evolutionary pressure. These mutations may significantly affect flocks that have received vaccinations as well as flocks with insufficient immunity in terms of viral immunity and disease dynamics. To assess this further, we investigated the efficacy of the heterologous inactivated LaSota or homologous genotype VII.1.1 vaccine for their protection against virulent NDV in chicken. Vaccine-induced immunity was evaluated based on the serology, and protection efficacy was assessed based on clinical signs, survival rates, histopathology, and viral shedding. Chickens vaccinated with the inactivated genotype VII.1.1 based vaccine showed 100% protection with a significant reduction in virus shedding, and ameliorated histopathology lesions compared to LaSota vaccinated chicks that showed 60% protection. These results revealed that the usage of NDV inactivated vaccine from the circulating field strains can successfully ameliorate the clinical outcome and virus pathobiology in vaccinated chicks and will serve as an effective vaccine against the threat posed by commonly circulating NDV strains in the poultry industry.

Genomic Diversity of a Globally Used, Live Attenuated Mycoplasma Vaccine.

The Mycoplasma synoviae live attenuated vaccine strain MS-H (Vaxsafe MS; Bioproperties Pty., Ltd., Australia) is commonly used around the world to prevent chronic infections caused by M. synoviae in birds and to minimize economic losses in the poultry industry. MS-H is a temperature-sensitive strain that is generated via the chemical mutagenesis of a virulent M. synoviae isolate, 86079/7NS. 32 single nucleotide polymorphisms have been found in the genome of MS-H compared to that of 86079/7NS, including 25 in predicted coding sequences (CDSs). There is limited information on the stability of these mutations in MS-H in vitro during the propagation of the vaccine manufacturing process or in vivo after the vaccination of chickens. Here, we performed a comparative analysis of MS-H genomes after in vitro and in vivo passages under different circumstances. Studying the dynamics of the MS-H population can provide insights into the factors that potentially affect the health of vaccinated birds. The genomes of 11 in vitro laboratory passages and 138 MS-H bird reisolates contained a total of 254 sequence variations. Of these, 39 variations associated with CDSs were detected in more than one genome (range = 2 to 62, median = 2.5), suggesting that these sequences are particularly prone to mutations. From the 25 CDSs containing previously characterized variations between MS-H and 86079/7NS, 7 were identified in the MS-H reisolates and progenies examined here. In conclusion, the MS-H genome contains individual regions that are prone to mutations that enable the restoration of the genotype or the phenotype of wild-type 86079/7NS in those regions. However, accumulated mutations in these regions are rare. IMPORTANCE Preventative measures, such as vaccination, are commonly used for the control of mycoplasmal infections in poultry. A live attenuated vaccine strain (Vaxsafe MS; MS-H; Bioproperties Pty. Ltd., Australia) is used for the prevention of disease caused by M. synoviae in many countries. However, information on the stability of previously characterized mutations in the MS-H genome is limited. In this study, we performed a comparative analysis of the whole-genome sequences of MS-H seeds used for vaccine manufacturing, commercial batches of the vaccine, cultures minimally passaged under small-scale laboratory and large-scale manufacturing conditions, MS-H reisolated from specific-pathogen-free (SPF) chickens that were vaccinated under controlled conditions, and MS-H reisolated from vaccinated commercial poultry flocks around the world. This study provides a comprehensive assessment of genome stability in MS-H after in vitro and in vivo passages under different circumstances and suggests that most of the mutations in the attenuated MS-H vaccine strain are stable.

Molecular phylodynamics of fowl adenovirus serotype 11 and 8b from inclusion body hepatitis outbreaks.

Fowl adenovirus (FAdV) serotypes are involved in a variety of clinical manifestations in poultry and has resulted in substantial economic loss to the poultry farmers. Despite the endemicity of Inclusion body hepatitis (IBH) in South Asian countries, including India, its etiology is not well studied. In western India, the rural poultry flocks obtained from the vaccinated parents were experiencing disease outbreaks with substantial economic losses due to heavy outbreaks and mortality. Therefore, the study was conducted to decipher the molecular epidemiology of the FAdV from field outbreaks in western India. A total of 37 commercial broiler poultry flocks and 29 village poultry flocks of western India were visited during 2019 to 2021. Out of these, 19.14% flocks showed incidence of IBH during the age of 15 to 35days. The mortality ranged from 3.3 percent to 55.28 percent. The samples were subjected for amplification of partial hexon gene covering loop 1 and loop 2. The results revealed 48.28% positivity by PCR. The sequence analysis identified 14 isolates as species D serotype 11 with 0.97 to 0.99% divergence and two as species E serotype 8b with 0.99% divergence. The FAdV-11 isolates showed amino acid substitutions D195N, T399A, N417S, and N496H. The amino acids I188 and N195 were conserved in FAdV-11. The molecular clock in Bayesian methods was used to determine most common ancestor. The isolates MH379249 and MH379248 were determined the most recent common ancestor for FAdV-11 and FAdV-8b isolates. The analysis suggested evolution of 10 FAdV-11 strains in 2012, and four FAdV-11 strains and two FAdV-8b strains in 2018.

A study of risk factors associated with Newcastle disease and molecular characterization of genotype XIII Newcastle disease virus in backyard and commercial poultry in Assam, India

The continuous emergence of Newcastle disease virus (NDV) poses a persistent threat to the poultry industry. Recent increasing outbreaks of NDV in the North East region of India have highlighted the need to closely monitor and analyze the potential risk factors for Newcastle disease (ND) outbreaks. In the present study, an attempt was made to genotype the circulating Newcastle disease virus (NDV) in the backyard and commercial poultry flocks in Assam, India. Sera samples from unvaccinated backyard poultry flocks and tissue samples of ND suspected cases were collected and tested for the presence of NDV antibodies using the Haemagglutination inhibition (HI) test. A total of seven NDV isolates were analyzed from different districts of Assam, India, both genotypically and pathotypically. All isolates were characterized as virulent, carrying 112RRKQRF117 amino acid residues at the cleavage site. As determined by phylogenetic analysis, the isolates clustered with members of genotype XIII of class II NDV. Further analysis of risk factors of ND occurrence was conducted through a questionnaire survey. All the results indicated an occurrence of genotype XIII of NDV in the farms with inadequate biosecurity and farming practices.

Mechanisms of biodiversity between Campylobacter sequence types in a flock of broiler-breeder chickens.

Commercial poultry flocks frequently harbor the dangerous bacterial pathogen Campylobacter. As exclusion efforts frequently fail, there is interest in potential ecologically informed solutions. A long‐term study of Campylobacter sequence types was used to investigate the competitive framework of the Campylobacter metacommunity and understand how multiple sequence types simultaneously co‐occur in a flock of chickens. A combination of matrix and patch‐occupancy models was used to estimate parameters describing the competition, transmission, and mortality of each sequence type. It was found that Campylobacter sequence types form a strong hierarchical framework within a flock of chickens and occupied a broad spectrum of transmission–mortality trade‐offs. Upon further investigation of how biodiversity is thus maintained within the flock, it was found that the demographic capabilities of Campylobacter, such as mortality and transmission, could not explain the broad biodiversity of sequence types seen, suggesting that external factors such as host‐bird health and seasonality are important elements in maintaining biodiversity of Campylobacter sequence types.

Pathobiology of highly pathogenic H5 avian influenza viruses in naturally infected Galliformes and Anseriformes in France during winter 2015\u20132016

In late 2015, an epizootic of Highly Pathogenic Avian Influenza (H5Nx) was registered in Southwestern France, including more than 70 outbreaks in commercial poultry flocks. Phylogenetic analyses suggested local emergence of H5 viruses which differed from A/goose/Guangdong/1/1996 clade 2.3.4.4b lineage and shared a unique polybasic cleavage site in their hemagglutinin protein. The present work provides an overview of the pathobiological picture associated with this epizootic in naturally infected chickens, guinea fowls and ducks. Upon necropsy examination, selected tissues were sampled for histopathology, immunohistochemistry and quantitative Real Time Polymerase Chain Reaction. In Galliformes, HPAIVs infection manifested as severe acute systemic vasculitis and parenchymal necrosis and was associated with endothelial expression of viral antigen. In ducks, lesions were mild and infrequent, with sparse antigenic detection in respiratory and digestive mucosae and leukocytes. Tissue quantifications of viral antigen and RNA were higher in chickens and guinea fowls compared to duck. Subsequently, recombinant HA (rHA) was generated from a H5 HPAIV isolated from an infected duck to investigate its glycan-binding affinity for avian mucosae. Glycan-binding analysis revealed strong affinity of rHA for 3’Sialyl-LacNAc and low affinity for Sialyl-LewisX, consistent with a duck-adapted virus similar to A/Duck/Mongolia/54/2001 (H5N2). K222R and S227R mutations on rHA sequence shifted affinity towards Sialyl-LewisX and led to an increased affinity for chicken mucosa, confirming the involvement of these two mutations in the glycan-binding specificity of the HA. Interestingly, the rHA glycan binding pattern of guinea fowl appeared intermediate between duck and chicken. The present study presents a unique pathobiological description of the H5 HPAIVs outbreaks that occurred in 2015–2016 in Southwestern France.

Serological investigation of low pathogenic avian influenza and Newcastle disease virus antibodies in Japanese quails, 30 village weavers and one laughing dove in two states of Nigeria

BackgroundIn spite of efforts to control avian influenza (AI) and Newcastle disease (ND) over decades, circulation of the viral causative agents among domestic and feral birds is considered implicated factors for the intermittent outbreaks of AI and ND among domesticated birds as well as commercial poultry flocks in Nigeria. In this study, sera from domestic (Japanese quails) and peri-domestic birds including laughing dove and village weavers were screened for antibodies to low pathogenic AI virus (LPAIV) and ND virus (NDV).MethodsA competitive ELISA was used to detect anti-AI virus antibodies in the sera of 101 unvaccinated Japanese quails, 30 village weavers and one laughing dove caught for human consumption in Oyo and Osun states, Nigeria. Hemagglutination-inhibiting (HI) antibodies against LPAIV were then detected in the ELISA-positive sera using H3N8, H5N2 and H9N7 subtype-specific antigens. Also, antibodies to NDV were detected and quantified in the sera using HI test.ResultsSeroprevalence of NDV antibodies from tested quail sera was 12.9% (13/101), while AI was 18.8% (19/101) with detection of anti-LPAIV H3N8, H5N2 and H9N7 antibodies. The laughing dove serum was positive for NDV and anti-LPAIV H9N7 antibodies while all sera from village weavers had no detectable LPAIV antibodies, but 26.7% (8/30) were positive for NDV antibodies.ConclusionsThis study provides serologic evidence of infection with LPAIV H3N8, H5N2 and H9N7 as well as Newcastle disease in domestic and peri-domestic birds in southwest Nigeria and highlights the potential role of these birds in the epidemiology of AI and ND.

Circulation and Molecular Characterization of Hemorrhagic Enteritis Virus in Commercial Turkey and Meat Chicken Flocks in Australia.

Currently, there is no available vaccine against hemorrhagic enteritis virus (HEV) in Australia. Although it is assumed that subclinical HEV infections occur and may be associated with an increase in colibacillosis in Australian commercial turkey flocks, the prevalence of infection with this virus in the country is largely unknown. The aims of this study were to determine the extent of HEV infection in commercial flocks in Australia and to investigate the diversity of Australian HEV strains. Serum and spleen samples were collected from breeder and grower turkeys and serum was collected from breeder and grower chickens by the two major poultry integrator companies in Australia. Of the turkey samples, 727/849 (86%) sera were positive for anti-HEV antibodies by ELISA. HEV DNA was detected in 215/278 (77%) spleen samples positive by PCR. Of the meat chicken sera, 115/144 (80%) samples were seropositive. Sequencing the whole genome of three HEV field isolates showed that the Australian strains are highly similar and cluster separately from strains from other geographic regions although several point mutations were shared with HEV strains considered to be virulent. In conclusion, HEV infection is ubiquitous in Australian commercial poultry flocks. The impact of the many genomic point mutations detected in Australian HEV strains on virus pathogenicity is unclear.