{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:41:24Z","timestamp":1785426084150,"version":"3.56.0"},"reference-count":51,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T00:00:00Z","timestamp":1776902400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key R&D Project","award":["2021YFD1800202"],"award-info":[{"award-number":["2021YFD1800202"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32373042"],"award-info":[{"award-number":["32373042"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32503006"],"award-info":[{"award-number":["32503006"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32373042"],"award-info":[{"award-number":["32373042"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"award":["32503006"],"award-info":[{"award-number":["32503006"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Basic Research Program of Jiangsu","award":["BK20250929"],"award-info":[{"award-number":["BK20250929"]}]},{"name":"Open Project Program of Jiangsu Key Laboratory of Zoonosis","award":["R2502"],"award-info":[{"award-number":["R2502"]}]},{"name":"Natural Science Foundation of Yangzhou","award":["YZ2025143"],"award-info":[{"award-number":["YZ2025143"]}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2025M773041"],"award-info":[{"award-number":["2025M773041"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"award":["2025M773041"],"award-info":[{"award-number":["2025M773041"]}],"id":[{"id":"https:\/\/ror.org\/0426zh255","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Jiangsu Province University Outstanding Science and Technology Innovation Team Project","award":["(2021) NO.1"],"award-info":[{"award-number":["(2021) NO.1"]}]},{"name":"Priority Academic Program Development of Jiangsu Higher Education"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>H5 subtype avian influenza virus (AIV) can infect both chickens and ducks, leading to substantial economic losses. Nevertheless, certain strains cause silent infections in ducks. In this study, a goose-origin clade 2.3.4.4h H5N6 AIV was isolated, which caused high mortality in mixed-gender white leghorn chickens but no deaths in mixed-gender mallard ducks. After independent serial in vitro passage in duck embryo fibroblasts (DEFs) and in vivo passage in specific-pathogen-free (SPF) ducks, the DEF-passage 10 (P10) virus induced markedly higher mortality rates and viral loads in SPF ducks compared to the DEF-P1 virus and the original parental virus prior to passage. Similarly, the in vivo-passaged P3 and P4 viruses exhibited significantly higher mortality rates than the P1 virus in SPF ducks, with 100% mortality and markedly increased viral titers in the organs. A whole-genome SNP analysis identified seven high-frequency mutations in the M1, NA and NS1 proteins. The NS1-F161L substitution virus exhibited significantly increased mortality rates, viral loads in multiple tissues, and a robustly induced innate immune response in ducks. Furthermore, dynamic evolutionary variations in the NS1 protein among global H5 avian influenza viruses revealed that the NS1-F161L substitution became dominant in clade 2.3.4.4b viruses in 2021 and subsequent years. Collectively, our findings demonstrate that host-driven adaptation can rapidly increase the pathogenicity of H5N6 AIVs in ducks and identify NS1-F161L as a critical virulence marker. These results offer novel insights relevant to the molecular surveillance, virulence prediction, and risk assessment of circulating H5 AIVs in waterfowl.<\/jats:p>","DOI":"10.3390\/v18050488","type":"journal-article","created":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T09:09:12Z","timestamp":1776935352000},"page":"488","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An NS1-F161L Substitution Determines Host-Driven Virulence Enhancement of H5N6 Avian Influenza Virus in Ducks"],"prefix":"10.3390","volume":"18","author":[{"given":"Yuwei","family":"Wu","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhifan","family":"Li","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7280-601X","authenticated-orcid":false,"given":"Nuo","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Animal Husbandry and Veterinary Medicine, Jiangsu Vocational College of Agriculture and Forestry, Jiangsu, Jurong 212400, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zijun","family":"Lu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yurui","family":"Dong","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kunlin","family":"Li","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Bian","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenzhi","family":"Huo","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1596-2168","authenticated-orcid":false,"given":"Tao","family":"Qin","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Interdisciplinary Center for Zoonoses and Biosafety, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sujuan","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Interdisciplinary Center for Zoonoses and Biosafety, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6042-3046","authenticated-orcid":false,"given":"Hui","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Interdisciplinary Center for Zoonoses and Biosafety, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6773-5149","authenticated-orcid":false,"given":"Daxin","family":"Peng","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Interdisciplinary Center for Zoonoses and Biosafety, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9924-6646","authenticated-orcid":false,"given":"Xiufan","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Research Centre of Engineering and Technology for Prevention and Control of Poultry Disease, Jiangsu, Yangzhou 225009, China"},{"name":"Jiangsu Interdisciplinary Center for Zoonoses and Biosafety, Yangzhou University, Jiangsu, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e00143-17","DOI":"10.1128\/JVI.00143-17","article-title":"Molecular Evolution and Emergence of H5N6 Avian Influenza Virus in Central China","volume":"91","author":"Du","year":"2017","journal-title":"J. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1016\/j.chom.2016.10.022","article-title":"Genesis, Evolution and Prevalence of H5N6 Avian Influenza Viruses in China","volume":"20","author":"Bi","year":"2016","journal-title":"Cell Host Microbe"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Li, H., Li, Q., Li, B., Guo, Y., Xing, J., Xu, Q., Liu, L., Zhang, J., Qi, W., and Jia, W. (2020). Continuous Reassortment of Clade 2.3.4.4 H5N6 Highly Pathogenetic Avian Influenza Viruses Demonstrating High Risk to Public Health. Pathogens, 9.","DOI":"10.3390\/pathogens9080670"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2956","DOI":"10.3201\/eid2612.201201","article-title":"Outbreaks of Highly Pathogenic Avian Influenza (H5N6) Virus Subclade 2.3.4.4h in Swans, Xinjiang, Western China, 2020","volume":"26","author":"Li","year":"2020","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2492","DOI":"10.3201\/eid2709.210819","article-title":"Highly Pathogenic Avian Influenza A(H5N6) Virus Clade 2.3.4.4h in Wild Birds and Live Poultry Markets, Bangladesh","volume":"27","author":"Turner","year":"2021","journal-title":"Emerg. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2218945","DOI":"10.1080\/22221751.2023.2218945","article-title":"Diverse infectivity, transmissibility, and pathobiology of clade 2.3.4.4 H5Nx highly pathogenic avian influenza viruses in chickens","volume":"12","author":"Kwon","year":"2023","journal-title":"Emerg. Microbes Infect."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"110971","DOI":"10.1016\/j.vetmic.2026.110971","article-title":"Isolation of reassortant H5N1 and H5N8 avian influenza viruses from co-infections and their host-dependent replication dynamics","volume":"316","author":"Wu","year":"2026","journal-title":"Vet. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"He, Z., Wang, X., Lin, Y., Feng, S., Huang, X., Zhao, L., Zhang, J., Ding, Y., Li, W., and Yuan, R. (2023). Genetic characteristics of waterfowl-origin H5N6 highly pathogenic avian influenza viruses and their pathogenesis in ducks and chickens. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1211355"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Wang, B., Su, Q., Luo, J., Li, M., Wu, Q., Chang, H., Du, J., Huang, C., Ma, J., and Han, S. (2021). Differences in Highly Pathogenic H5N6 Avian Influenza Viral Pathogenicity and Inflammatory Response in Chickens and Ducks. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.593202"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.virol.2019.01.016","article-title":"Different pathogenicity of two strains of clade 2.3.4.4c H5N6 highly pathogenic avian influenza viruses bearing different PA and NS gene in domestic ducks","volume":"530","author":"Kwon","year":"2019","journal-title":"Virology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2250065","DOI":"10.1080\/21505594.2023.2250065","article-title":"Virulence and transmission characteristics of clade 2.3.4.4b H5N6 subtype avian influenza viruses possessing different internal gene constellations","volume":"14","author":"Zhao","year":"2023","journal-title":"Virulence"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1227","DOI":"10.1111\/tbed.13141","article-title":"Comparative pathogenicity of H5N6 subtype highly pathogenic avian influenza viruses in chicken, Pekin duck and Muscovy duck","volume":"66","author":"Uchida","year":"2019","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4439","DOI":"10.1128\/JVI.75.9.4439-4443.2001","article-title":"Generation of a highly pathogenic avian influenza A virus from an avirulent field isolate by passaging in chickens","volume":"75","author":"Ito","year":"2001","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1007\/s11427-009-0115-3","article-title":"Stable non-synonymous substitutions on NS gene (NS1 and NS2 proteins) of Qinghai Lake H5N1 influenza virus (Clade 2.2) after successive passages in Muscovy ducks","volume":"52","author":"Song","year":"2009","journal-title":"Sci. China Ser. C Life Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2455","DOI":"10.1007\/s00705-015-2504-1","article-title":"Serial passage in ducks of a low-pathogenic avian influenza virus isolated from a chicken reveals a high mutation rate in the hemagglutinin that is likely due to selection in the host","volume":"160","author":"Ridenour","year":"2015","journal-title":"Arch. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Li, Y., Chen, S., Zhang, X., Fu, Q., Zhang, Z., Shi, S., Zhu, Y., Gu, M., Peng, D., and Liu, X. (2014). A 20-amino-acid deletion in the neuraminidase stalk and a five-amino-acid deletion in the NS1 protein both contribute to the pathogenicity of H5N1 avian influenza viruses in mallard ducks. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0095539"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11063","DOI":"10.1128\/JVI.00760-13","article-title":"The PA and HA gene-mediated high viral load and intense innate immune response in the brain contribute to the high pathogenicity of H5N1 avian influenza virus in mallard ducks","volume":"87","author":"Hu","year":"2013","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2180","DOI":"10.1128\/JVI.01975-10","article-title":"The PA protein directly contributes to the virulence of H5N1 avian influenza viruses in domestic ducks","volume":"85","author":"Song","year":"2011","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Nao, N., Kajihara, M., Manzoor, R., Maruyama, J., Yoshida, R., Muramatsu, M., Miyamoto, H., Igarashi, M., Eguchi, N., and Sato, M. (2015). A Single Amino Acid in the M1 Protein Responsible for the Different Pathogenic Potentials of H5N1 Highly Pathogenic Avian Influenza Virus Strains. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0137989"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"W232","DOI":"10.1093\/nar\/gkw256","article-title":"W-IQ-TREE: A fast online phylogenetic tool for maximum likelihood analysis","volume":"44","author":"Trifinopoulos","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e01551-18","DOI":"10.1128\/JVI.01551-18","article-title":"Mutation of Influenza A Virus PA-X Decreases Pathogenicity in Chicken Embryos and Can Increase the Yield of Reassortant Candidate Vaccine Viruses","volume":"93","author":"Hussain","year":"2019","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6288","DOI":"10.1038\/s41467-022-33909-2","article-title":"The influenza virus PB2 protein evades antiviral innate immunity by inhibiting JAK1\/STAT signalling","volume":"13","author":"Yang","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e00289-20","DOI":"10.1128\/JVI.00289-20","article-title":"The Microbiota Contributes to the Control of Highly Pathogenic H5N9 Influenza Virus Replication in Ducks","volume":"94","author":"Figueroa","year":"2020","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Yang, H., Wang, Y., Jin, S., Pang, Q., Shan, A., and Feng, X. (2022). Dietary resveratrol alleviated lipopolysaccharide-induced ileitis through Nrf2 and NF-\u03baB signalling pathways in ducks (Anas platyrhynchos). J. Anim. Physiol. Anim. Nutr., 106.","DOI":"10.1111\/jpn.13657"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Bessi\u00e8re, P., Figueroa, T., Coggon, A., Foret-Lucas, C., Houffschmitt, A., Fusade-Boyer, M., Dupr\u00e9, G., Gu\u00e9rin, J.L., Delverdier, M., and Volmer, R. (2022). Opposite Outcomes of the Within-Host Competition between High- and Low-Pathogenic H5N8 Avian Influenza Viruses in Chickens Compared to Ducks. J.Virol., 96.","DOI":"10.1128\/JVI.01366-21"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"441","DOI":"10.4161\/viru.25710","article-title":"Genetic changes that accompanied shifts of low pathogenic avian influenza viruses toward higher pathogenicity in poultry","volume":"4","author":"Abdelwhab","year":"2013","journal-title":"Virulence"},{"key":"ref_28","first-page":"103979","article-title":"Evolution of H9N2 low pathogenic avian influenza virus during passages in chickens","volume":"75","author":"Giza","year":"2019","journal-title":"Infect. Genet. Evol. J. Mol. Epidemiol. Evol. Genet. Infect. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1186\/s12985-025-02793-9","article-title":"Dynamic adaptation mutations and pathogenic characterization of a mouse-adapted seasonal human H3N2 influenza virus","volume":"22","author":"Zhang","year":"2025","journal-title":"Virol. J."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Dai, M., Zhu, S., An, Z., You, B., Li, Z., Yao, Y., Nair, V., and Liao, M. (2023). Dissection of key factors correlating with H5N1 avian influenza virus driven inflammatory lung injury of chicken identified by single-cell analysis. PLoS Pathog., 19.","DOI":"10.1371\/journal.ppat.1011685"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Evseev, D., and Magor, K.E. (2019). Innate Immune Responses to Avian Influenza Viruses in Ducks and Chickens. Vet. Sci., 6.","DOI":"10.3390\/vetsci6010005"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"101203","DOI":"10.1016\/j.lanmic.2025.101203","article-title":"Duck-origin H5N6 avian influenza threatens public health: A challenge for poultry vaccination in China","volume":"6","author":"Wang","year":"2025","journal-title":"Lancet Microbe"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Huang, J., Wu, S., Wu, W., Liang, Y., Zhuang, H., Ye, Z., Qu, X., Liao, M., and Jiao, P. (2021). The Biological Characteristics of Novel H5N6 Highly Pathogenic Avian Influenza Virus and Its Pathogenesis in Ducks. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.628545"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Park, M.J., Cha, R.M., Kye, S.J., Lee, Y.N., Kim, N.Y., Baek, Y.G., Heo, G.B., Sagong, M., Lee, K.N., and Lee, Y.J. (2021). Pathogenicity of H5N8 High Pathogenicity Avian Influenza Virus in Chickens and Ducks from South Korea in 2020\u20132021. Viruses, 13.","DOI":"10.3390\/v13101903"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Jia, D., Rahbar, R., Chan, R.W., Lee, S.M., Chan, M.C., Wang, B.X., Baker, D.P., Sun, B., Peiris, J.S., and Nicholls, J.M. (2010). Influenza virus non-structural protein 1 (NS1) disrupts interferon signaling. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0013927"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"6849","DOI":"10.1128\/JVI.02323-08","article-title":"The NS1 protein of a human influenza virus inhibits type I interferon production and the induction of antiviral responses in primary human dendritic and respiratory epithelial cells","volume":"83","author":"Haye","year":"2009","journal-title":"J. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Abd El-Hamid, H.S., Shafi, M.E., Albaqami, N.M., Ellakany, H.F., Abdelaziz, N.M., Abdelaziz, M.N., Abd El-Hack, M.E., Taha, A.E., Alanazi, K.M., and Elbestawy, A.R. (2020). Sequence analysis and pathogenicity of Avian Orthoavulavirus 1 strains isolated from poultry flocks during 2015\u20132019. BMC Vet. Res., 16.","DOI":"10.1186\/s12917-020-02470-9"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3864","DOI":"10.1038\/ncomms4864","article-title":"Pathogenic potential of interferon \u03b1\u03b2 in acute influenza infection","volume":"5","author":"Davidson","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e00776-22","DOI":"10.1128\/jvi.00776-22","article-title":"Avian Influenza NS1 Proteins Inhibit Human, but Not Duck, RIG-I Ubiquitination and Interferon Signaling","volume":"96","author":"Evseev","year":"2022","journal-title":"J. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"11566","DOI":"10.1128\/JVI.74.24.11566-11573.2000","article-title":"Influenza A virus NS1 protein prevents activation of NF-kappaB and induction of alpha\/beta interferon","volume":"74","author":"Wang","year":"2000","journal-title":"J. Virol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e01081-17","DOI":"10.1128\/JVI.01081-17","article-title":"Structure-Guided Functional Annotation of the Influenza A Virus NS1 Protein Reveals Dynamic Evolution of the p85\u03b2-Binding Site during Circulation in Humans","volume":"91","author":"Lopes","year":"2017","journal-title":"J. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"eadu4909","DOI":"10.1126\/sciadv.adu4909","article-title":"Spatiotemporal reconstruction of the North American A(H5N1) outbreak reveals successive lineage replacements by descendant reassortants","volume":"11","author":"Signore","year":"2025","journal-title":"Sci. Adv."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"269","DOI":"10.4142\/jvs.2017.18.S1.269","article-title":"Evolution, global spread, and pathogenicity of highly pathogenic avian influenza H5Nx clade 2.3.4.4","volume":"18","author":"Lee","year":"2017","journal-title":"J. Vet. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"637","DOI":"10.3201\/eid2304.161866","article-title":"Highly Pathogenic Avian Influenza A(H5N8) Virus in Wild Migratory Birds, Qinghai Lake, China","volume":"23","author":"Li","year":"2017","journal-title":"Emerg. Infect. Dis."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1080\/22221751.2021.1872355","article-title":"Emergence and spread of novel H5N8, H5N5 and H5N1 clade 2.3.4.4 highly pathogenic avian influenza in 2020","volume":"10","author":"Lewis","year":"2021","journal-title":"Emerg. Microbes Infect."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1007\/s11427-021-2025-y","article-title":"Genetic and biological characteristics of the globally circulating H5N8 avian influenza viruses and the protective efficacy offered by the poultry vaccine currently used in China","volume":"65","author":"Cui","year":"2022","journal-title":"Sci. China. Life Sci."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Khalil, A.M., Fujimoto, Y., Kojima, I., Esaki, M., Ri, K., Masatani, T., Matsui, T., and Ozawa, M. (2021). Genetic Characterization of H5N8 Highly Pathogenic Avian Influenza Viruses Isolated from Falcated Ducks and Environmental Water in Japan in November 2020. Pathogens, 10.","DOI":"10.3390\/pathogens10020171"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"104005","DOI":"10.1016\/j.isci.2022.104005","article-title":"Evolutionary pressures rendered by animal husbandry practices for avian influenza viruses to adapt to humans","volume":"25","author":"Caetano","year":"2022","journal-title":"iScience"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"7952","DOI":"10.1038\/ncomms8952","article-title":"Phylodynamics of H1N1\/2009 influenza reveals the transition from host adaptation to immune-driven selection","volume":"6","author":"Su","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"C8","DOI":"10.1016\/j.vaccine.2008.04.011","article-title":"Vaccination and antigenic drift in influenza","volume":"26","author":"Boni","year":"2008","journal-title":"Vaccine"},{"key":"ref_51","first-page":"100377","article-title":"Genomic signatures and host adaptation of H5N1 clade 2.3.4.4b: A call for global surveillance and multi-target antiviral strategies","volume":"8","author":"Zhang","year":"2025","journal-title":"Curr. Res. Microb. Sci."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/18\/5\/488\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T09:12:57Z","timestamp":1776935577000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/18\/5\/488"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,23]]},"references-count":51,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["v18050488"],"URL":"https:\/\/doi.org\/10.3390\/v18050488","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,23]]}}}