{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T09:02:55Z","timestamp":1779699775418,"version":"3.53.1"},"reference-count":30,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T00:00:00Z","timestamp":1779667200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"2026 Hebei Provincial Project for the Introduction of Foreign Intelligence","award":["360-0806-YBN-5V7K"],"award-info":[{"award-number":["360-0806-YBN-5V7K"]}]},{"name":"Special Project of Basic Scientific Research Funds for Colleges and Universities in Hebei Province","award":["2025JK019"],"award-info":[{"award-number":["2025JK019"]}]},{"DOI":"10.13039\/501100015286","name":"Key Research and Development Program of Hebei Province","doi-asserted-by":"publisher","award":["213777109D"],"award-info":[{"award-number":["213777109D"]}],"id":[{"id":"10.13039\/501100015286","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Scientific Research Project of the General Administration of Customs","award":["2024HK119"],"award-info":[{"award-number":["2024HK119"]}]},{"name":"Scientific Research Project of the General Administration of Customs","award":["2024HK264"],"award-info":[{"award-number":["2024HK264"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>During 2022\u20132024, a highly pathogenic avian influenza virus (HPAIV) H5N1 strain, designated A\/Seagull\/Hebei\/qhd6\/2024 (H5N1), was isolated from migratory birds in Beidaihe National Wetland Park, North China. Phylogenetic analyses revealed that its hemagglutinin (HA) gene belongs to the 2.3.4.4b clade, while the neuraminidase (NA) gene and internal genes clustered with strains originating from multiple continents, consistent with a transcontinental reassortment event. The virus also exhibited 90.1\u201398.1% nucleotide homology with human-derived H5N1 isolates. Molecular characterization identified key virulence-associated mutations, including the classic HPAIV HA cleavage site, HA-T160A (associated with enhanced human receptor-binding capacity), and NA-I117T (potentially linked to drug resistance). BALB\/c mouse infection experiments confirmed systemic replication and high pathogenicity of strain qhd6, with a 50% lethal dose (LD50) of 0.95 log10EID50\/mL. Antigenic analysis revealed good cross-reactivity with the widely used H5-Re14 vaccine strain. This study reports the identification, in Beidaihe National Wetland Park, of an HPAIV H5N1 strain whose genetic characteristics suggest intercontinental reassortment and indicate cross-species transmission risk. It clarifies the genetic characteristics and pathogenicity of this strain, providing an important theoretical and practical basis for precise surveillance, risk early warning, and comprehensive prevention and control of AIV at migratory bird stopover sites in North China.<\/jats:p>","DOI":"10.3390\/v18060595","type":"journal-article","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T08:06:26Z","timestamp":1779696386000},"page":"595","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Clade 2.3.4.4b H5N1 HPAIV from Migratory Birds in Beidaihe Wetland, North China"],"prefix":"10.3390","volume":"18","author":[{"given":"Yiyang","family":"Zhang","sequence":"first","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoli","family":"Bai","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenhui","family":"Nie","sequence":"additional","affiliation":[{"name":"Qinhuangdao Bureau of Agriculture and Rural Affairs, Qinhuangdao 066001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yufei","family":"Guo","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Shan","sequence":"additional","affiliation":[{"name":"Qinhuangdao Customs Technology Center, Qinhuangdao 066099, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanxia","family":"Xiao","sequence":"additional","affiliation":[{"name":"Qinhuangdao Customs Technology Center, Qinhuangdao 066099, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoqing","family":"Zhang","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-3924-9422","authenticated-orcid":false,"given":"Shuaiyu","family":"Jiang","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongmei","family":"Su","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng","family":"Chang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Special Animal Epidemic Disease, Ministry of Agriculture and Rural Affairs, Jilin Provincial International Cooperation Key Laboratory for Science and Technology Innovation of Special Animal and Plants, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural, Changchun 130112, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongsheng","family":"Liu","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shunli","family":"Yang","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanbing","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Tian","sequence":"additional","affiliation":[{"name":"The Science and Technology Research Center of China Customs, Beijing 100026, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boru","family":"Zhang","sequence":"additional","affiliation":[{"name":"Qinhuangdao Forestry Bureau, Qinhuangdao 066001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Liang","sequence":"additional","affiliation":[{"name":"Qinhuangdao Forestry Bureau, Qinhuangdao 066001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexei D.","family":"Zaberezhny","sequence":"additional","affiliation":[{"name":"Federal State Budgetary Institution All-Russian Research and Technological Institute of Biological Industry (VNITIBP),17,bldg.1,Village of the Biokombinat, Losino-Petrovsky Urban District, Moscow Oblast 141142, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunkai","family":"Qian","sequence":"additional","affiliation":[{"name":"Qinhuangdao Customs Technology Center, Qinhuangdao 066099, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Zhang","sequence":"additional","affiliation":[{"name":"Hebei Province Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaorui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Special Animal Epidemic Disease, Ministry of Agriculture and Rural Affairs, Jilin Provincial International Cooperation Key Laboratory for Science and Technology Innovation of Special Animal and Plants, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural, Changchun 130112, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"101526","DOI":"10.1016\/j.psj.2021.101526","article-title":"Evidence of Mycoplasma spp. Transmission by Migratory Wild Geese","volume":"101","author":"Tomczyk","year":"2022","journal-title":"Poult. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1186\/s40168-022-01246-7","article-title":"Virome in the Cloaca of Wild and Breeding Birds Revealed a Diversity of Significant Viruses","volume":"10","author":"Shan","year":"2022","journal-title":"Microbiome"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"102947","DOI":"10.1016\/j.psj.2023.102947","article-title":"First Isolation and Genomic Characterization of Avian Reovirus from Black Swans (Cygnus atratus) in China","volume":"102","author":"Zhu","year":"2023","journal-title":"Poult. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"104430","DOI":"10.1016\/j.psj.2024.104430","article-title":"Research Note: Study on the Characteristics of Salmonella typhimurium Derived from Larus vegae","volume":"103","author":"Guo","year":"2024","journal-title":"Poult. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/nature01333","article-title":"Fingerprints of Global Warming on Wild Animals and Plants","volume":"421","author":"Root","year":"2003","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Verhagen, J.H., Fouchier, R.A.M., and Lewis, N. (2021). Highly Pathogenic Avian Influenza Viruses at the Wild\u2013Domestic Bird Interface in Europe: Future Directions for Research and Surveillance. Viruses, 13.","DOI":"10.3390\/v13020212"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2298","DOI":"10.3201\/eid2911.230332","article-title":"Human Salmonellosis Outbreak Linked to Salmonella Typhimurium Epidemic in Wild Songbirds, United States, 2020\u20132021","volume":"29","author":"Patel","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Nishiura, H., Kumagai, A., Mine, J., Takadate, Y., Sakuma, S., Tsunekuni, R., Uchida, Y., and Miyazawa, K. (2024). Phylogenetic and Pathogenic Analysis of H5N1 and H5N6 High Pathogenicity Avian Influenza Virus Isolated from Poultry Farms (Layer and Broiler Chickens) in Japan in the 2023\/2024 Season. Viruses, 16.","DOI":"10.3390\/v16121956"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Yang, L., Wang, R., Liu, Q., Shah, T., Zhou, J., Zhao, W., Wang, Y., Deng, L., and Wang, B. (2025). Genomic Characterization and Phylogenetic Analysis of Five Avian Influenza H5N1 Subtypes from Wild Anser indicus in Yunnan, China. Vet. Sci., 12.","DOI":"10.3390\/vetsci12030280"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Chrzastek, K., Lieber, C.M., and Plemper, R.K. (2025). H5N1 Clade 2.3.4.4b: Evolution, Global Spread, and Host Range Expansion. Pathogens, 14.","DOI":"10.3390\/pathogens14090929"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1111\/zph.12149","article-title":"Prevalence, Characterization and Antibiotic Resistance of Salmonella Isolates in Large Corvid Species of Europe and North America between 2010 and 2013","volume":"62","author":"Janecko","year":"2015","journal-title":"Zoonoses Public Health"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"157189","DOI":"10.1016\/j.scitotenv.2022.157189","article-title":"Eurasian griffon vultures Carry Widespread Antimicrobial Resistant Salmonella and Campylobacter of Public Health Concern","volume":"844","author":"Espunyes","year":"2022","journal-title":"Sci. Total Environ."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Amado, M.A., Shin, H., Sanz, V., Lentino, M., Mart\u00ednez, L.M., Contreras, M., Michelangeli, F., and Dom\u00ednguez-Bello, M.G. (2018). Comparison of Gizzard and Intestinal Microbiota of Wild Neotropical Birds. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0194857"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Laviad-Shitrit, S., Izhaki, I., Lalzar, M., and Halpern, M. (2019). Comparative Analysis of Intestine Microbiota of Four Wild Waterbird Species. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.01911"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.3201\/eid3007.240508","article-title":"Highly Pathogenic Avian Influenza A(H5N1) Clade 2.3.4.4b Virus Infection in Domestic Dairy Cattle and Cats, United States, 2024","volume":"30","author":"Burrough","year":"2024","journal-title":"Emerg. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e02542-24","DOI":"10.1128\/mbio.02542-24","article-title":"Avian Influenza A (H5N1) Virus in Dairy Cattle: Origin, Evolution, and Cross-Species Transmission","volume":"15","author":"Mostafa","year":"2024","journal-title":"mBio"},{"key":"ref_17","unstructured":"(2020). Diagnostic Techniques for Highly Pathogenic Avian Influenza (Standard No. GB\/T 18936-2020)."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.vetmic.2019.06.005","article-title":"A Novel I117T Substitution in Neuraminidase of Highly Pathogenic Avian Influenza H5N1 Virus Conferring Reduced Susceptibility to Oseltamivir and Zanamivir","volume":"235","author":"Kode","year":"2019","journal-title":"Vet. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3947","DOI":"10.1128\/JVI.03328-14","article-title":"Identification of PB2 Mutations Responsible for the Efficient Replication of H5N1 Influenza Viruses in Human Lung Epithelial Cells","volume":"89","author":"Yamaji","year":"2015","journal-title":"J. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e01930-19","DOI":"10.1128\/JVI.01930-19","article-title":"A Single Amino Acid at Position 431 of the PB2 Protein Determines the Virulence of H1N1 Swine Influenza Viruses in Mice","volume":"94","author":"Xu","year":"2020","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1099\/vir.0.017459-0","article-title":"Emergence of H5N1 Avian Influenza Viruses with Reduced Sensitivity to Neuraminidase Inhibitors and Novel Reassortants in Lao People\u2019s Democratic Republic","volume":"91","author":"Seiler","year":"2010","journal-title":"J. Gen. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1093\/infdis\/jiq010","article-title":"Generation and Characterization of Recombinant Pandemic Influenza A(H1N1) Viruses Resistant to Neuraminidase Inhibitors","volume":"203","author":"Pizzorno","year":"2011","journal-title":"J. Infect. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.chom.2018.08.006","article-title":"Rapid Evolution of H7N9 Highly Pathogenic Viruses that Emerged in China in 2017","volume":"24","author":"Shi","year":"2018","journal-title":"Cell Host Microbe"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e01717-21","DOI":"10.1128\/JVI.01717-21","article-title":"Emergence, Evolution, and Pathogenicity of Influenza A(H7N4) Virus in Shorebirds in China","volume":"96","author":"Chai","year":"2022","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1080\/22221751.2022.2088407","article-title":"Global Dissemination of H5N1 Influenza Viruses Bearing the Clade 2.3.4.4b HA Gene and Biologic Analysis of the Ones Detected in China","volume":"11","author":"Cui","year":"2022","journal-title":"Emerg. Microbes Infect."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2380421","DOI":"10.1080\/22221751.2024.2380421","article-title":"Genomic Characterization of Highly Pathogenic Avian Influenza A H5N1 Virus Newly Emerged in Dairy Cattle","volume":"13","author":"Hu","year":"2024","journal-title":"Emerg. Microbes Infect."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Huo, C., Xiao, K., Zhang, S., Tang, Y., Wang, M., Qi, P., Xiao, J., Tian, H., and Hu, Y. (2018). H5N1 Influenza a Virus Replicates Productively in Pancreatic Cells and Induces Apoptosis and Pro-Inflammatory Cytokine Response. Front. Cell. Infect. Microbiol., 8.","DOI":"10.3389\/fcimb.2018.00386"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2502005","DOI":"10.1080\/22221751.2025.2502005","article-title":"Unique Phenomenon of H5 Highly Pathogenic Avian Influenza Virus in China: Co-Circulation of Clade 2.3.4.4b H5N1 and H5N6 Results in Diversity of H5 Virus","volume":"14","author":"Zhang","year":"2025","journal-title":"Emerg. Microbes Infect."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e02209-24","DOI":"10.1128\/jvi.02209-24","article-title":"Highly Pathogenic Avian Influenza H5N1: History, Current Situation, and Outlook","volume":"99","author":"Krammer","year":"2025","journal-title":"J. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"fuae014","DOI":"10.1093\/femsre\/fuae014","article-title":"Epitopes in the HA and NA of H5 and H7 Avian Influenza Viruses that Are Important for Antigenic Drift","volume":"48","author":"Luczo","year":"2024","journal-title":"FEMS Microbiol. Rev."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/18\/6\/595\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T08:09:10Z","timestamp":1779696550000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/18\/6\/595"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,25]]},"references-count":30,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["v18060595"],"URL":"https:\/\/doi.org\/10.3390\/v18060595","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,25]]}}}