,Tabulated individual data points for data reported in the associated publication: Spackman E, Suarez DL, Lee CW, Pantin-Jackwood MJ, Lee SA, Youk S, Ibrahim S. Efficacy of inactivated and RNA particle vaccines against a North American Clade 2.3.4.4b H5 highly pathogenic avian influenza virus in chickens. Vaccine. 2023 Nov 30;41(49):7369-7376. doi: 10.1016/j.vaccine.2023.10.070. Epub 2023 Nov 4. PMID: 37932132.,The highly pathogenic avian influenza virus (HPAIV) isolate A/turkey/Indiana/22-003707-003/2022 H5N1 (TK/IN/22) and A/Gyrfalcon/Washington/41088/2014 H5N8 (GF/WA/14) isolate were each propagated and titrated in embryonating specific pathogen free (SPF) chicken eggs using standard procedures and titers were determined using the Reed-Muench method.,An in-house vaccine was produced by de novo synthesizing the HA gene of TK/IN/22 that was modified to be low pathogenic (LP) and placing it in a PR8 backbone using rg methods as described . The vaccine (SEP-22-N9) contained 6 genes from PR8 and a de novo synthesized N9 NA from A/blue winged teal/Wyoming/AH0099021/2016 (H7N9). The rg virus was inactivated by treatment with 0.1% beta-propiolactone. Vaccines were produced with Montanide ISA 71 VG (Seppic Inc., Fairfield, NJ) adjuvant at ambient temperature in a L5M-A high shear mixer (Silverson Machines, Inc., East Longmeadow, MA) for 30sec at 1,000rpm, then for 3min at 4,000rpm using an emulsifying screen in accordance with the adjuvant manufacturer’s instructions.,Sham vaccine was prepared in-house using sterile phosphate buffered saline as described above.,Commercial vaccines were supplied by the manufacturers. The commercial inactivated vaccine (1057.R1 serial 590088) (rgH5N1) (Zoetis Inc., Parsippany, NJ) was produced with the GF/WA/14 (clade 2.3.4.4c HA gene) and the remaining 7 gene segments including the NA from PR8 (1). The Sequivity vaccine (serial V040122NCF) (RP) (Merck and Co. Inc., Rahway, NJ) is an updated version of their replication restricted alphavirus vector vaccine that expresses the TK/IN/22 H5 HA (modified to be low pathogenic LP).,Three-week-old, mixed sex, SPF white leghorn chickens (Gallus gallus domesticus) were obtained from in-house flocks and were randomly assigned to vaccine groups.,All vaccines were administered by the subcutaneous route at the nape of the neck. Commercial vaccines were given at the volumes instructed by the manufacturer (0.5ml each). In-house vaccine was given at a dose of 512 hemagglutination units per bird in 0.5ml. Three weeks post vaccination chickens were challenged with 6.7 log10 50% egg infectious doses (EID50) of TK/IN/22 in 0.1ml by the intrachoanal route.,Oropharyngeal (OP) and cloacal (CL) swabs were collected from all birds at 2-, 4-, and 7-days post challenge (DPC). Swabs were also collected from dead and euthanized sham vaccinates at 1DPC.,To evaluate antibody-based DIVA-VI tests, blood for serum was collected from the RP and SEP-22-N9 vaccinated groups at 7, 10 and 14DPC because the SEP-22-N9 vaccine does not elicit antibodies to N1 and the RP vaccine does not elicit antibodies to the N1 or NP proteins.,Mortality and morbidity were recorded for 14DPC after which time the remaining birds were euthanized. If birds were severely lethargic or had neurological signs they were euthanized and were counted as mortality at the next observation time for mean death time calculations.,To determine if there was a difference in antibody levels based on the order of vaccination with the RP vaccine and an inactivated vaccine, groups of 20 chickens (hatch-mates of the chickens in the challenge study) were given one dose of each vaccine three weeks apart (Supplementary Table 1). The first dose was administered at three weeks of age using the RP or SEP-22-N9 vaccine as described above. Then a second dose of either the same vaccine or the other vaccine was administered three weeks later (six weeks of age). All birds were bled for serum three weeks after the second vaccination (nine

,The global outbreak of clade 2.3.4.4b H5N1 highly pathogenic avian influenza (HPAI) virus has caused tremendous losses in poultry. Although turkeys are a smaller sector in poultry production compared to chickens, they tend to be affected more severely by HPAI virus because they can usually be infected with a lower dose of influenza A virus than chickens (i.e., they are more susceptible). Exposure to non-replicating proteins may help control HPAI, however data with turkeys are somewhat limited regarding how well they work and approaches to modifying surveillance have not been developed. Here, an H5N9 non-replicating protein comprised of a clade 2.3.4.4b H5 hemagglutinin from A/turkey/Indiana/22-003707-003/2022 (TK/IN/22) and a North American wild bird lineage N9 was evaluated in commercial broad breasted white turkeys by challenge to live virus. Turkeys were divided into three groups, where each group was exposed to the non-replicating protein once at 3 (3wk), 7 (7wk), or 9 (9wk) weeks of age. All birds were challenged at 10 weeks of age with TK/IN/22 HPAIV. There was 100% survival in all groups except the sham exposure group which had 100% mortality. A significant decrease in viral shedding was observed in all exposed groups compared to the shams, although the 9wk group shed significantly higher quantities by the cloacal route at seven days post challenge (DPC) compared to the 3wk group. The neuraminidase inhibition-enzyme linked lectin assay (NI-ELLA) was used as a serological test that was able to detect antibody in birds that had been infected after exposure and challenge based on antibodies to the NA protein of the challenge virus (N1 NA) in serum collected 7, 10 and 14DPC. Between 50 and 90% of turkeys, depending on age at exposure, were positive by NI-ELLA at 7DPC and 100% were positive at 14DPC regardless of age at exposure.,

,Two commercially available vaccines based on the recombinant herpes virus of turkeys (rHVT) vector were tested against a recent North American clade 2.3.4.4b HPAI virus isolate: A/turkey/Indiana/22-003707-003/2022 H5N1 in specific pathogen free white leghorn (WL) chickens and commercial broiler chickens. One rHVT-H5 vaccine encodes a hemagglutinin (HA) gene designed by the computationally optimized broadly reactive antigen method (COBRA-HVT vaccine). The other encodes an HA gene of a clade 2.2 virus (2.2-HVT vaccine). There was 100% survival of both breeds in the COBRA-HVT vaccinated groups and in the 2.2-HVT vaccinated groups there was 94.8% and 90% survival of the WL and broilers respectively. Compared to the 2.2-HVT vaccinated groups, WL in the COBRA-HVT vaccinated group shed significantly lower mean viral titers by the cloacal route and broilers shed significantly lower titers by the oropharyngeal route than broilers. Virus titers detected in oral and cloacal swabs were otherwise similar among both vaccine groups and chicken breeds. To assess antibody-based tests to identify birds that have been infected after vaccination (DIVA-VI), sera collected after the challenge were tested with enzyme-linked lectin assay-neuraminidase inhibition (ELLA-NI) for N1 neuraminidase antibody detection and by commercial ELISA for detection of antibodies to the NP protein. As early as 7 days post challenge (DPC) 100% of the chickens were positive by ELLA-NI. ELISA was less sensitive with a maximum of 75% positive at 10DPC in broilers vaccinated with 2.2-HVT. Both vaccines provided protection from challenge to both breeds of chickens and ELLA-NI was sensitive at identifying antibodies to the challenge virus therefore should be evaluated further for DIVA-VI.,Methods,Viruses. All procedures using infectious material were reviewed and approved by the Institutional Biosafety Committee of US National Poultry Research Center (USNPRC), US Department of Agriculture-Agricultural Research Service, Athens, GA. The HPAI virus isolate A/turkey/Indiana/22-003707-003/2022 H5N1 (TK/IN/22) was provided by Dr. Mia Torchetti, National Veterinary Services Laboratories, US Department of Agriculture-Animal and Plant Health Inspection Service, Ames, IA. The A/Vietnam/1203/2004 H5N1 HPAI virus (Viet/04), A/Whooper Swan/Mongolia/244/2005 H5N1 (WS/Mongolia/05) HPAI virus, and A/Flycatcher/CA/14875-1/1994 H7N1 low pathogenic avian influenza virus isolates were provided by the repository at the USNPRC. Virus isolates were propagated and titrated in SPF embryonating chicken eggs using standard procedures [1]. Titers were determined using the Reed-Muench method [2].,Vaccines. Two commercial rHVT-H5 vaccines were selected because they are licensed in the US (and may be licensed elsewhere) and were supplied by the manufacturers: 2.2-HVT (Vectormune HVT AIV, Ceva Animal Health LLC, Lenexa, KS) (serial 395-134); and COBRA-HVT (Vaxxitek HVT+IBD+H5, Boehringer-Ingelheim Animal Health USA, Ridgefield, CT) (serial EW003). The amino acid similarity between the vaccine antigens and the challenge virus HA1 was 91.7% (COBRA-HVT) and 91.2% (2.2-HVT).,Challenge study design. All animal work was reviewed and approved by the USNPRC Institutional Animal Care and Use Committee. Mixed sex, SPF WL chickens (Gallus gallus domesticus) were obtained at hatch from in-house flocks. Broiler chicken eggs were obtained from a commercial hatchery at 18 days of incubation prior to administration of any in ovo vaccines and were hatched on-site. All birds were randomly assigned to vaccine groups based on breed. Vaccine groups are shown in Table 1. All vaccines were prepared and administered on the day of hatch by the subcutaneous route at the nape of the neck in accordance with the manufacturer’s instructions (0.2ml per chicken). Serum was collected from all chickens 25 days post vaccination to evaluate the antibody response to the vaccines.,Four weeks post vaccination (four weeks of age) chickens were

Males and females from resistant Fayoumi and susceptible Leghorn chicken lines were either challenged with a lentogenic strain of Newcastle Disease virus or given a mock infection at 3 weeks of age. The lung transcriptomes generated by RNA-sequencing were studied using contrasts across the challenged and non-challenged birds, the two lines, and three time points (2,6, and 10 days post-infection) using Weighted Gene Co-expression Network Analysis (WGNCA). The data can be retrieved by navigating to https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-5859/. Click the Download button to access the Sample and data relationship dataset file.

These data are a result of an experiment to examine the effect of imidacloprid on chicken immunity and behavior. There are therefore several sets of physiological data. Citation information for this dataset can be found in the EDG's Metadata Reference Information section and Data.gov's References section.

,Data are the individual group values for oral and cloacal virus shedding and antibody titers for reach treatment group from: Mo et al., The pathogenicity and transmission of live bird market H2N2 avian influenza viruses in chickens, Pekin ducks, and guinea fowl. Vet Mic 260:109180, 2021. https://doi.org/10.1016/j.vetmic.2021.109180,Methods: Six H2N2 low pathogenic avian influenza viruses from US LBMs were selected based on recency and to represent the different genotypes present in the live birds markets during the time period (i.e., the presence or absence of a NA stalk deletion): A/duck/PA/14-030488-5/2014 (Dk/PA/14), A/chicken/NY/16-032621-2/2016 (Ck/NY/16), A/chicken/CT/17-008911-4/2017 (Ck/CT/17), A/chicken/NY/18-002471-4/2018 (CK/NY/02471/18), A/chicken/NY/18-042097-3/2018 (Ck/NY/042097/18) and A/chicken/NY/19-012787-1/2019 (Ck/NY/19). Isolates were evaluated in White Leghorn chickens (Gallus gallus), guinea fowl (Numida meleagris) and Pekin ducks (Anas platyrhynchos). Chickens and guinea fowl were challenged at 4 weeks of age and Pekin ducks were challenged at 2 weeks of age with 6log10 of virus by the intra-choanal route. “Contact” birds, which were hatch-mates of the inoculated birds, were co-housed with the inoculated birds 24hrs post inoculation to evaluate transmission. Viral loads in OP and CL swabs collected at 2, 4, 7, 10, and 14 days post inoculation were determined by quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR). RNA was extracted from swabs using the MagMAX96 Viral RNA Isolation Kit (Thermo Fisher Scientific, Waltham, MA) and the KingFisher Flex Magnetic Particle Processing System (Thermo Fisher Scientific), with an additional wash step to remove inhibitors (Das et al., 2009). The qRT‐PCR for AIV detection was conducted based on the standard USDA M gene AIV qRT‐PCR procedure (Spackman et al., 2002) using an Applied Biosystems® 7500 Fast Real‐Time PCR system (Thermo Fisher Scientific). Cycle threshold (Ct) values were determined by the 7500 Fast Software v2.3. For relative quantification, Ct values were converted to titer equivalents based on the standard curve method (Larionov et al., 2005). Values were established from ten-fold dilutions of the same titrated stock of the virus used to challenge the birds. The limit of detection was determined to be 0.8Log10 per reaction. Serological testing for antibodies to the virus utilized the hemagglutination inhibition (HI) assays using homologous antigens were performed to quantify antibody responses with serum collected from chickens, guinea fowl and Pekin ducks at 14 dpi based on the standard protocol (OIE, 2019). HI titers were reported as reciprocal log2 titers, and titers greater than 3 log2 (1:8) were considered positive.,,

To determine the gender impact on the immune response of chickens, the mRNA was isolated and sequenced from the lungs of 48 chickens of 2 lines as three time-points post-infection (2,6, and 10 days post-infection), and in two treatment groups. The data can be retrieved by navigating to https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-5859/.

These data were collected as part of a field trial to test the efficacy of a sylvatic plague vaccine. Treatment and control sites were selected randomly from the available sites at each location. Site pairs were a minimum of 20 acres, (with a few exceptions). Prairie dog trapping took place a minimum of two weeks post-baiting and trapping procedures were approved by the NWHC Animal Care and Use Committee as well as individual states as required.

These data were collected as part of a field trial to test the efficacy of a sylvatic plague vaccine. Treatment and control sites were selected randomly from the available sites at each location. Site pairs were a minimum of 20 acres, (with a few exceptions). Prairie dog trapping took place a minimum of two weeks post-baiting and trapping procedures were approved by the NWHC Animal Care and Use Committee as well as individual states as required.

,Treatment groups of ducks were exposed to different virus doses (2, 4, 6 log10 50% egg infectious doses) and by different routes (contact or intrachoanal). The experimental setting was a laboratory with animal care as approved by the institutional animal care and use committee as appropriate for the species and age of bird. Data are the virus titers shed by the oral and cloacal route for individual mallard ducks exposed to H5N1 highly pathogenic avian influenza virus by day post exposure. Samples were collected through 11 days post exposure. Virus titer equivalents were determined by quantitative real-time RT-PCR. Serological data are serum antibody titers to the challenge virus as determined by hemagglutination inhibition assay (reciprocal of the log2 dilution) determined with serum collected 10 or 11 days post exposure.,,