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The interferon-induced proteins with tetratricopeptide repeats (IFITs) protein family mediates antiviral effects by inhibiting translation initiation, cell proliferation, and migration in the interferon (IFN) dependent innate immune system. Several members of this family, including IFIT1, IFIT2, IFIT3 and IFIT5, have been heavily studied in mammals. Avian species contain only one family member, IFIT5, and little is known about the role of this protein in birds. In this study, duck IFIT5 (duIFIT5) full-length mRNA was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of the cDNA ends (RACE). Based on the sequence obtained, we performed a series of bioinformatics analyses, and found that duIFIT5 was most similar to homologs in other avian species. Also, duIFIT5 contained eight conserved TPR motifs and two conserved multi-domains (TPR_11 and TPR_12). Finally, we used duck hepatitis virus type 1 (DHV-1) and polyriboinosinicpolyribocytidylic acid (poly (I:C)) as a pathogen or a pathogen-associated molecular pattern induction to infect three-day-old domestic ducklings. The liver and spleen were collected to detect the change in duIFIT5 transcript level upon infection by quantitative real-time PCR (qRT-PCR). DuIFIT5 expression rapidly increased after DHV-1 infection and maintained a high level, while the transcripts of duIFIT5 peaked at 8h after poly (I:C) infection and then returned to normal. Taken together, these results provide a greater understanding of avian IFIT5.

The VP1 protein of duck hepatitis A virus (DHAV) is a major structural protein that induces neutralizing antibodies in ducks; however, B-cell epitopes on the VP1 protein of duck hepatitis A genotype 1 virus (DHAV-1) have not been characterized. To characterize B-cell epitopes on VP1, we used the monoclonal antibody (mAb) 2D10 against Escherichia coli-expressed VP1 of DHAV-1. In vitro, mAb 2D10 neutralized DHAV-1 virus. By using an array of overlapping 12-mer peptides, we found that mAb 2D10 recognized phages displaying peptides with the consensus motif LPAPTS. Sequence alignment showed that the epitope 173LPAPTS178 is highly conserved among the DHAV-1 genotypes. Moreover, the six amino acid peptide LPAPTS was proven to be the minimal unit of the epitope with maximal binding activity to mAb 2D10. DHAV-1-positive duck serum reacted with the epitope in dot blotting assay, revealing the importance of the six amino acids of the epitope for antibody-epitope binding. Competitive inhibition assays of mAb 2D10 binding to synthetic LPAPTS peptides and truncated VP1 protein fragments, detected by Western blotting, also verify that LPAPTS was the VP1 epitope. We identified LPAPTS as a VP1-specific linear B-cell epitope recognized by the neutralizing mAb 2D10. Our findings have potential applications in the development of diagnostic techniques and epitope-based marker vaccines against DHAV-1.

A Newcastle disease virus (NDV)-vectored vaccine expressing clade 2.3.4.4b H5 Hemagglutinin was developed and assessed for efficacy against H5N1 highly pathogenic avian influenza (HPAI) in specific pathogen-free (SPF) chickens, broilers, and domestic ducks. In SPF chickens, the live recombinant NDV-vectored vaccine, rK148/22-H5, achieved complete survival against HPAI and NDV challenges and significantly reduced viral shedding. Notably, the live rK148/22-H5 vaccine conferred good clinical protection in broilers despite the presence of maternally derived antibodies. Good clinical protection was observed in domestic ducks, with decreased viral shedding. It demonstrated complete survival and reduced cloacal viral shedding when used as an inactivated vaccine from SPF chickens. The rK148/22-H5 vaccine is potentially a viable and supportive option for biosecurity measure, effectively protecting in chickens against the deadly clade 2.3.4.4b H5 HPAI and NDV infections. Furthermore, it aligns with the strategy of Differentiating Infected from Vaccinated Animals (DIVA).

The duck (Anas platyrhynchos) is one of the principal natural hosts of influenza A viruses. We present the duck genome sequence and perform deep transcriptome analyses to investigate immune-related genes. Our data indicate that the duck possesses a contractive immune gene repertoire, as in chicken and zebra finch, and this repertoire has been shaped through lineage-specific duplications. We identify genes that are responsive to influenza A viruses using the lung transcriptomes of control ducks and ones that were infected with either a highly pathogenic (A/duck/Hubei/49/05) or a weakly pathogenic (A/goose/Hubei/65/05) H5N1 virus. Further, we show how the duck's defense mechanisms against influenza infection have been optimized through the diversification of its β-defensin and butyrophilin-like repertoires. These analyses, in combination with the genomic and transcriptomic data, provide a resource for characterizing the interaction between host and influenza viruses.

We recently reported the development of a successful post-exposure combination antiviral and “prime-boost” vaccination strategy using the duck hepatitis B virus (DHBV) model of human hepatitis B virus infection. The current study aimed to simplify the vaccination strategy and to test the post-exposure efficacy of combination therapy with the Bristol-Myers Squibb antiviral drug, entecavir (ETV) and either a single dose of DHBV DNA vaccines on day 0 post-infection (p.i.) or a single dose of recombinant fowlpoxvirus (rFPV–DHBV) vaccines on day 7 p.i. Whilst untreated control ducks infected with an equal dose of DHBV all developed persistent and wide spread DHBV infection of the liver, ducks treated with ETV combined with either the DHBV DNA vaccines on day 0 p.i. or the rFPV–DHBV vaccines on day 7 p.i. had no detectable DHBV-infected hepatocytes by day 14 p.i. and were protected from the development of persistent DHBV infection.

Faced with increasingly disastrous avian influenza crises, and considering the associated zoonotic risk, France launched a vaccination campaign against highly pathogenic avian influenza on duck farms with more than 250 ducks in October 2023. In the context of this paradigm shift in the fight against HPAI, our study was designed to (i) provide an overview of the vaccination campaign over the first six-month period and (ii) evaluate the level of assumed vaccine protection at population scale through a simple modelling approach – using actual duck population dynamics, data related to the recommended vaccination protocol, and biological data on vaccination-induced humoral responses obtained from experimental studies - and real-world data on how vaccination was actually implemented in the field between 1 October 2023 and 31 March 2024, corresponding to the level of compliance with the vaccination protocol. Vaccination compliance appeared satisfactory, with high vaccination coverage at flock level (more than 95% of duck flocks being vaccinated at least once during the studied period). Ages at vaccination and time between doses were in line with the vaccination protocol. Regarding the assessment of the level of assumed vaccine protection, both theoretical model and actual vaccination data indicated that, at any given time point, maximum 40–50% of duck batches were considered fully protected, through either the second dose or the booster, while 30–40% of the vaccinated flocks were either considered only partially protected (primary protection through the first dose) or their protection was seen to wane six weeks after the second dose. Our work can inform the testing and optimisation of vaccination strategies, and complementing our data-driven approach with serological monitoring, evaluation of vaccine efficacy, or spatio-temporal modelling of coverage across territorial scales would be pivotal to fully capture the interplay between vaccination, population dynamics, stakeholder decision-making, and epidemiological dynamics. •We analysed data from the French avian influenza vaccination plan on duck farms.•More than 95% of duck flocks were vaccinated during the study period.•Ages at and time between vaccinations were in line with the recommended protocol.•Modelling and field data showed that less than half of flocks had full protection.•Our work could be used to test protocols and optimize vaccination strategies.

Ducks and wild waterfowl perpetuate all strains of influenza viruses in nature. In their natural host, influenza viruses typically cause asymptomatic infection and little pathology. Ducks are often resistant to influenza viruses capable of killing chickens. Here, we show that the influenza virus sensor, RIG-I, is present in ducks and plays a role in clearing an influenza infection. We show evidence suggesting that RIG-I may be absent in chickens, providing a plausible explanation for their increased susceptibility to influenza viruses compared with ducks. RIG-I detects RNA ligands derived from uncapped viral transcripts and initiates the IFN response. In this study, we show that the chicken embryonic fibroblast cell line, DF-1, cannot respond to a RIG-I ligand. However, transfection of duck RIG-I into DF-1 cells rescues the detection of ligand and induces IFN-β promoter activity. Additionally, DF-1 cells expressing duck RIG-I have an augmented IFN response resulting in decreased influenza replication after challenge with either low or highly pathogenic avian influenza virus. Implicating RIG-I in the antiviral response to an infection in vivo, we found that RIG-I expression is induced 200 fold, early in an innate immune response in ducks challenged with the H5N1 virus A/Vietnam/1203/04. Finding this natural disease resistance gene in ducks opens the possibility of increasing influenza resistance through creation of a transgenic chicken.

The Florida Mottled Duck ( Anas fulvigula fulvigula ) is a unique subspecies of waterfowl whose range is limited to peninsular Florida, USA. As an endemic subspecies, Florida Mottled Ducks face numerous conservation stressors, such as habitat conversion and hybridization with non-native Mallards ( Anas platyrhynchos ). In addition to these numerous stressors, Mottled Ducks are also contending with emerging and/or geographically expanding waterborne pathogens such as Vibrio spp., due to the effects of climate change. However, even given their conservation needs, little is known with respect to the health, physiology, and the immunity of wild Mottled Ducks in Florida. Given this lack of data, we performed health assessments of Mottled Ducks in the Central Florida area. Specifically, we examined the humoral innate immune system, i.e., the plasma of Mottled Ducks in response to a common but extraneous pathogen: Escherichia coli strain American Type Culture Collection (ATCC) number 8739. We utilized a bactericidal assay (“bacterial killing assay” or BKA) commonly used in eco-immunology, to provide insight into the bactericidal capacities of captured Florida Mottled Ducks. We statistically tested the BKA capacity, or microbial killing ability, of 23 Mottled Ducks in response to age and whole blood lead levels (Pb). We found that there was no statistically significant relationship between the covariates we measured and Mottled Duck BKA capacity against E. coli . However, the variability we observed in the BKA capacity of this subspecies warrants further research into additional physiological and ecological covariates coupled with potential immune stressors that Florida Mottled Ducks may be contending with.

Pasteurella multocida (P. multocida), a pathogenic bacterium known to induce duck cholera, stands as a significant contributor to bacterial diseases afflicting the duck industry, causing substantial annual economic losses on a global scale. In this study, the genes encoding the lipoproteins PlpE of P. multocida strain PMWSG-4 was cloned, inserted into the pBAD-ClyA vector, and the recombinant outer membrane vesicles (OMVs) fused with PlpE antigen of P. multocida was expressed by Escherichia coli (E. coli). Ducks immunized with OMV-PlpE had significantly (P < 0.001) increased production of antigen-specific antibodies. Moreover, at 28 days post-immunization, the expression of genes associated with immune response, including interleukin (IL)-2, IL-4, IL-10, and interferon (IFN)-γ in the spleen tissue of immunized ducks were significantly (P < 0.001) up-regulated compared to unimmunized ducks in the control group. And the active serum had significant bactericidal effects against the PMWSG-4 strain (P < 0.001). The protective efficacy of the vaccines was evaluated by leg muscle challenge with 20 LD50 doses of P. multocida, with the recombinant OMV-PlpE conferring 100 % protection. Histopathological examination and tissue bacterial load detection revealed that OMV-PlpE mitigated tissue damage and bacterial colonization to a statistically significant extent (P < 0.001). These findings serve as a valuable reference for the development of vaccines against P. multocida.

Phosphatase and tensin homolog (PTEN) is a well-established tumor suppressor gene that antagonizes the PI3K/AKT signaling pathway and plays a critical role in regulating both innate and adaptive immune responses. However, its function in avian species, particularly in ducks, remains largely unexplored. In this study, the full-length cDNA of duck PTEN ( PTEN) was cloned from the spleen of healthy Cherry Valley ducks. Sequence alignment and phylogenetic analysis were performed to evaluate its structural conservation and evolutionary relationships. The tissue distribution of duPTEN mRNA was examined using quantitative real-time PCR (qRT-PCR). Furthermore, duPTEN expression was assessed in the spleen, lung, and brain at 1, 3, and 5 days post-infection (dpi) following exposure to Duck Tembusu Virus (DTMUV), Duck Plague Virus (DPV), and Novel Duck Reovirus (NDRV). Sequence analysis demonstrated that PTEN shares a high degree of conservation with PTEN from other avian species, displaying 100% identity with sequences from and . qRT-PCR results revealed that PTEN is ubiquitously expressed across multiple tissues, with the highest expression observed in the brain. Upon DTMUV and DPV infection, duPTEN expression was significantly upregulated in the spleen and lung but downregulated in the brain. In contrast, NDRV infection led to consistent downregulation of PTEN across all three tissues. This study is the first to characterize the molecular cloning, tissue-specific expression, and virus-induced regulation of PTEN in ducks. The findings suggest that PTEN plays a role in the host immune response to diverse viral infections, highlighting its potential involvement in the regulation of antiviral innate immunity in avian species.