{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:54:28Z","timestamp":1791467668586,"version":"4.3.4"},"reference-count":47,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Dutch Ministry of Agriculture, Nature and Food Quality","award":["WOT-01-003-096, KB-37-003-047-WBVR"],"award-info":[{"award-number":["WOT-01-003-096, KB-37-003-047-WBVR"]}],"id":[{"id":"https:\/\/ror.org\/03b1hdw57","id-type":"ROR","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["RGPIN\/3774-2022"],"award-info":[{"award-number":["RGPIN\/3774-2022"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]},{"name":"RIKEN Center for Interdisciplinary Theoretical and Mathematical Sciences","id":[{"id":"https:\/\/ror.org\/022hvg611","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"abstract":"ABSTRACT<\/jats:title>\n \n \n \n The introduction, spread, and subsequent detection of highly pathogenic avian influenza viruses (HPAIVs) in poultry are influenced by the virus replication fitness, transmission fitness, and virulence. Quantifying these fitness parameters is important for implementing surveillance and control measures for poultry. This study employs an avian\n in vitro<\/jats:italic>\n model using primary chicken embryo fibroblast and duck embryo fibroblast to identify cellular correlates of fitness for a reference panel of eight dominant, highly pathogenic avian influenza clade 2.3.4.4 virus genotypes (H5N1, H5N8, and H5N6) isolated between 2014 and 2024. Because HPAIVs are pantropic in poultry, the use of primary fibroblasts could provide a robust systemic model to assess virus fitness in different poultry species. By integrating longitudinal infectious virus titers and cytopathogenicity data with a mathematical model, which delineates cell populations into susceptible, latent, infectious, and dead compartments, we disentangled \u201ctraditional\u201d virological parameters such as peak virus titer and time to 50% cell death from mechanistic parameters, including the infecting time (\n \n \n \n \n \n \n \n \n t<\/mml:mi>\n \n \n i<\/mml:mi>\n n<\/mml:mi>\n f<\/mml:mi>\n <\/mml:mrow>\n <\/mml:mrow>\n <\/mml:msub>\n <\/mml:mstyle>\n <\/mml:mrow>\n <\/mml:mstyle>\n <\/mml:math>\n <\/jats:alternatives>\n <\/jats:inline-formula>\n ), generation time (\n \n \n \n \n \n \n \n \n t<\/mml:mi>\n \n \n g<\/mml:mi>\n e<\/mml:mi>\n n<\/mml:mi>\n <\/mml:mrow>\n <\/mml:mrow>\n <\/mml:msub>\n <\/mml:mstyle>\n <\/mml:mrow>\n <\/mml:mstyle>\n <\/mml:math>\n <\/jats:alternatives>\n <\/jats:inline-formula>\n ), and basic reproduction number (\n R<\/jats:italic>\n 0<\/jats:sub>\n ). Our results confirm host-specific fitness characteristics consistent with\n in vivo<\/jats:italic>\n observations. This study underscores the potential of integrating avian\n in vitro<\/jats:italic>\n models with mathematical modeling to provide high-resolution characterization of genetically similar viruses, building toward rapid risk assessments of novel HPAIVs.\n <\/jats:p>\n \n IMPORTANCE<\/jats:title>\n \n Highly pathogenic avian influenza (HPAI) outbreaks in poultry have resulted in the culling of millions of animals over the past decade. The introduction and spread of HPAI in poultry are driven by the ability of the virus to replicate, transmit, and cause disease, which varies among different poultry species. These viral fitness traits can change quickly through mutations or reassortment, leading to new variants with an altered risk of introduction and transmission. In this study, we integrated an\n in vitro<\/jats:italic>\n and mathematical model to assess the viral fitness of genetically similar viruses and the risk they represent for two important poultry species. This approach offers a rapid and scalable alternative to traditional animal models, aligning with the 3R\u2019s principle and supporting rapid assessment of the viral fitness of emerging virus variants for poultry. This approach can be easily expanded to other species, including humans, and is suitable to assess the viral fitness of all viral pathogens.\n <\/jats:p>\n <\/jats:sec>\n <\/jats:sec>","DOI":"10.1128\/spectrum.00843-26","type":"journal-article","created":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:01:06Z","timestamp":1791464466000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Mathematical modeling of\n in vitro<\/i>\n replication dynamics for multiple highly pathogenic avian influenza clade 2.3.4.4 viruses in chicken and duck cells"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6958-7925","authenticated-orcid":true,"given":"Luca","family":"Bordes","sequence":"first","affiliation":[{"name":"Department of Virology, Wageningen Bioveterinary Research","place":["Lelystad, the Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1893-0559","authenticated-orcid":false,"given":"Peter","family":"Hobbelen","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Bioinformatics & Animal Models, Wageningen Bioveterinary Research","place":["Lelystad, the Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Blokker","sequence":"additional","affiliation":[{"name":"Department of Virology, Wageningen Bioveterinary Research","place":["Lelystad, the Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7498-8002","authenticated-orcid":false,"given":"Wim H. M.","family":"van der Poel","sequence":"additional","affiliation":[{"name":"Department of Virology, Wageningen Bioveterinary Research","place":["Lelystad, the Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0599-0069","authenticated-orcid":false,"given":"Catherine A. A.","family":"Beauchemin","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05g13zd79","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Physics, Toronto Metropolitan University","place":["Toronto, Canada"]},{"name":"RIKEN Center for Interdisciplinary Theoretical and Mathematical Sciences (iTHEMS), RIKEN","place":["Wako, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6756-4431","authenticated-orcid":false,"given":"Nancy","family":"Beerens","sequence":"additional","affiliation":[{"name":"Department of Virology, Wageningen Bioveterinary Research","place":["Lelystad, the Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7889-5380","authenticated-orcid":true,"given":"Jos\u00e9 L.","family":"Gonzales","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Bioinformatics & Animal Models, Wageningen Bioveterinary Research","place":["Lelystad, the Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,10,8]]},"reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.00609-22"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268818001292"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.2903\/j.efsa.2022.7597"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.3390\/v13020212"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1404.071016"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41426-018-0070-9"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2022.03.010"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2024.2399970"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0966-842X(97)81772-6"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2020.1868274"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00955-21"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00149-22"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.3390\/v15020265"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cimid.2023.102039"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.3390\/v11090812"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1615\/critrevimmunol.v30.i3.60"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.4161\/viru.2.5.17267"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.046219-0"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2458-11-S1-S7"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.epidem.2020.100406"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01623-05"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep24154"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.3390\/v11060501"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.es2015.20.26.21174"},{"key":"e_1_3_4_26_2","unstructured":"IZSVe. 2024. FluMut. https:\/\/github.com\/izsvenezie-virology\/FluMut."},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1099\/jgv.0.002007"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0192645"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1098\/rsif.2008.0172"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.epidem.2019.100368"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1214\/17-STS618"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1002730"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.12748"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.envsoft.2019.07.007"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0014767"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.01536-22"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.3390\/v16040563"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.3390\/v15091909"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1637\/11882-042718-Reg.1"},{"key":"e_1_3_4_40_2","unstructured":"Germeraad E Velkers F Jong M Gonzales J Wit J Stegeman J Beerens N. 2023. Transmissiestudie met vier vaccins tegen H5N1 hoogpathogeen vogelgriepvirus. Wageningen Bioveterinary Research"},{"key":"e_1_3_4_41_2","unstructured":"ANSES resume of experimental evaluation of transmission among vaccinated ducks. 2024. https:\/\/www.anses.fr\/en\/system\/files\/Resume_Expe_Transmission.pdf (02-10-2024."},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.14675"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep39505"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-025-02714-w"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.psj.2025.104921"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.3390\/v14010165"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2458-11-S1-S10"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0126115"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.00843-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.00843-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:01:08Z","timestamp":1791464468000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.00843-26"}},"subtitle":[],"editor":[{"given":"Tao","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,10,8]]},"references-count":47,"alternative-id":["10.1128\/spectrum.00843-26"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.00843-26","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,8]]},"assertion":[{"value":"2026-03-09","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-26","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-10-08","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00843-26"}}