{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T19:10:34Z","timestamp":1789758634903,"version":"4.0.1"},"update-to":[{"DOI":"10.1371\/journal.ppat.1014604","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000}}],"reference-count":74,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T00:00:00Z","timestamp":1788912000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>Seasonal influenza viruses continue to pose a significant threat to human health. As influenza viruses exhibit sustained genetic drift, it is imperative that animal studies utilize challenge strains that reflect contemporary, currently circulating viruses when evaluating pathogenicity, viral tropism, transmissibility, and antiviral sensitivity to better inform public health responses. Ferrets are considered the gold-standard small animal model for assessing currently circulating influenza viruses. Seasonal influenza A(H1N1)pdm09 viruses replicate well in both the upper and lower respiratory tract of ferrets, providing an important model for developing improved vaccination and therapeutic strategies; however, many of these studies have relied on a 2009 virus isolate. Utilising representative influenza A(H1N1)pdm09 virus strains from 2009 to 2022, we explored virus replication kinetics and lung pathogenesis in ferrets following intranasal inoculation with these contemporary strains. Our results revealed strain specific differences, with greater lung viral loads and pathogenesis following inoculation with A\/Sydney\/5\/2021 compared to other strains. Efficient transmissibility of A\/Sydney\/5\/2021 virus to na\u00efve recipients was also observed following both contact and airborne exposure to infected donor ferrets. To refine this updated model, we performed side-by-side evaluation of oseltamivir antiviral efficacy following traditional intranasal or aerosol inhalation influenza challenges. Pre-treatment with oseltamivir demonstrated greater reductions in viral shedding from the upper respiratory tract than post-infection treatment of ferrets infected by aerosol inhalation, while the intranasal route showed reduced oseltamivir efficacy independent of the timing of antiviral treatment. These findings provide the basis for using an updated A(H1N1)pdm09 challenge virus for ferret studies as an alternative to the commonly used, but now less relevant 2009 early pandemic viruses. It also highlights how different methods of virus inoculation can influence outcomes of ferret antiviral studies, with an aerosol challenge model able to demonstrate differences between therapeutic and pre-treatment schedules, which were not apparent with an intranasal challenge model.<\/jats:p>","DOI":"10.1371\/journal.ppat.1014604","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T17:43:02Z","timestamp":1788975782000},"page":"e1014604","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Updated A(H1N1)pdm09 influenza virus ferret infection model permits refined antiviral assessment using aerosol inhalation challenge"],"prefix":"10.1371","volume":"22","author":[{"given":"Harry L.","family":"Stannard","sequence":"first","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, at The Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"]},{"name":"University of Melbourne, at the Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"],"department":["Department of Microbiology and Immunology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"Brock","sequence":"additional","affiliation":[{"name":"National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention","place":["Atlanta, Georgia, United States of America"],"department":["Influenza Division"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangjie","family":"Sun","sequence":"additional","affiliation":[{"name":"National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention","place":["Atlanta, Georgia, United States of America"],"department":["Influenza Division"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna A.","family":"Pulit-Penaloza","sequence":"additional","affiliation":[{"name":"National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention","place":["Atlanta, Georgia, United States of America"],"department":["Influenza Division"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Troy J.","family":"Kieran","sequence":"additional","affiliation":[{"name":"National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention","place":["Atlanta, Georgia, United States of America"],"department":["Influenza Division"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashwin","family":"Muraleetharan","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, at The Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clyde","family":"Dapat","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, at The Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Smitha Rose","family":"Georgy","sequence":"additional","affiliation":[{"name":"The University of Melbourne","place":["Melbourne, Victoria, Australia"],"department":["Section of Anatomic Pathology, Melbourne Veterinary School, Faculty of Science"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taronna R.","family":"Maines","sequence":"additional","affiliation":[{"name":"National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention","place":["Atlanta, Georgia, United States of America"],"department":["Influenza Division"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian G.","family":"Barr","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, at The Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"]},{"name":"University of Melbourne, at the Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"],"department":["Department of Microbiology and Immunology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick C.","family":"Reading","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, at The Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"]},{"name":"University of Melbourne, at the Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"],"department":["Department of Microbiology and Immunology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saira","family":"Hussain","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, at The Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"]},{"name":"University of Melbourne, at the Peter Doherty Institute for Infection and Immunity","place":["Melbourne, Victoria, Australia"],"department":["Department of Microbiology and Immunology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0755-7368","authenticated-orcid":true,"given":"Jessica A.","family":"Belser","sequence":"additional","affiliation":[{"name":"National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention","place":["Atlanta, Georgia, United States of America"],"department":["Influenza Division"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,9,9]]},"reference":[{"issue":"10127","key":"ppat.1014604.ref001","doi-asserted-by":"crossref","first-page":"1285","DOI":"10.1016\/S0140-6736(17)33293-2","article-title":"Estimates of global seasonal influenza-associated respiratory mortality: a modelling study","volume":"391","author":"AD Iuliano","year":"2018","journal-title":"Lancet"},{"key":"ppat.1014604.ref002","unstructured":"WHO. Up to 650 000 people die of respiratory diseases linked to seasonal flu each year. https:\/\/www.who.int\/news\/item\/13-12-2017-up-to-650-000-people-die-of-respiratory-diseases-linked-to-seasonal-flu-each-year 2017. Accessed 2021 August."},{"issue":"10","key":"ppat.1014604.ref003","doi-asserted-by":"crossref","first-page":"2269","DOI":"10.3390\/v2102269","article-title":"Neuraminidase inhibitor susceptibility testing in human influenza viruses: a laboratory surveillance perspective","volume":"2","author":"M Okomo-Adhiambo","year":"2010","journal-title":"Viruses"},{"key":"ppat.1014604.ref004","doi-asserted-by":"crossref","first-page":"106217","DOI":"10.1016\/j.antiviral.2025.106217","article-title":"Global update on the susceptibilities of influenza viruses to neuraminidase inhibitors and the cap-dependent endonuclease inhibitor baloxavir, 2020-2023","volume":"241","author":"S Hussain","year":"2025","journal-title":"Antiviral Research"},{"issue":"3","key":"ppat.1014604.ref005","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1128\/MMBR.00022-16","article-title":"Complexities in ferret influenza virus pathogenesis and transmission models","volume":"80","author":"JA Belser","year":"2016","journal-title":"Microbiology and Molecular Biology Reviews"},{"issue":"1","key":"ppat.1014604.ref006","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.ajpath.2019.09.017","article-title":"A Guide for the Use of the Ferret Model for Influenza Virus Infection","volume":"190","author":"JA Belser","year":"2020","journal-title":"Am J Pathol"},{"issue":"5939","key":"ppat.1014604.ref007","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1126\/science.1177238","article-title":"Transmission and pathogenesis of swine-origin 2009 A(H1N1) influenza viruses in ferrets and mice","volume":"325","author":"TR Maines","year":"2009","journal-title":"Science"},{"issue":"5939","key":"ppat.1014604.ref008","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1126\/science.1177127","article-title":"Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets","volume":"325","author":"VJ Munster","year":"2009","journal-title":"Science"},{"issue":"8","key":"ppat.1014604.ref009","article-title":"Comparison of temporal and spatial dynamics of seasonal H3N2, pandemic H1N1 and highly pathogenic avian influenza H5N1 virus infections in ferrets","volume":"7","author":"JMA van den Brand","year":"2012","journal-title":"PLoS One"},{"issue":"3","key":"ppat.1014604.ref010","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1177\/0300985811429812","article-title":"Modification of the ferret model for pneumonia from seasonal human influenza A virus infection","volume":"49","author":"JMA van den Brand","year":"2012","journal-title":"Vet Pathol"},{"issue":"1","key":"ppat.1014604.ref011","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.virol.2012.06.002","article-title":"Surface glycoproteins of influenza A H3N2 virus modulate virus replication in the respiratory tract of ferrets","volume":"432","author":"X Cheng","year":"2012","journal-title":"Virology"},{"key":"ppat.1014604.ref012","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.1099\/vir.0.064352-0","article-title":"Pathogenic influenza B virus in the ferret model establishes lower respiratory tract infection","volume":"95","author":"SSH Huang","year":"2014","journal-title":"J Gen Virol"},{"issue":"3","key":"ppat.1014604.ref013","doi-asserted-by":"crossref","first-page":"365","DOI":"10.3390\/ph16030365","article-title":"Preclinical Study of ZSP1273, a Potent Antiviral Inhibitor of Cap Binding to the PB2 Subunit of Influenza A Polymerase","volume":"16","author":"X Chen","year":"2023","journal-title":"Pharmaceuticals (Basel)"},{"issue":"4","key":"ppat.1014604.ref014","doi-asserted-by":"crossref","DOI":"10.1128\/mbio.01056-22","article-title":"Effect of Baloxavir and Oseltamivir in Combination on Infection with Influenza Viruses with PA\/I38T or PA\/E23K Substitutions in the Ferret Model","volume":"13","author":"P Koszalka","year":"2022","journal-title":"mBio"},{"issue":"4","key":"ppat.1014604.ref015","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1011342","article-title":"4\u2019-Fluorouridine mitigates lethal infection with pandemic human and highly pathogenic avian influenza viruses","volume":"19","author":"CM Lieber","year":"2023","journal-title":"PLoS Pathog"},{"issue":"2","key":"ppat.1014604.ref016","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1011993","article-title":"Influenza A virus resistance to 4\u2019-fluorouridine coincides with viral attenuation in vitro and in vivo","volume":"20","author":"CM Lieber","year":"2024","journal-title":"PLoS Pathog"},{"key":"ppat.1014604.ref017","doi-asserted-by":"crossref","first-page":"104751","DOI":"10.1016\/j.antiviral.2020.104751","article-title":"Prophylaxis of ferrets with nitazoxanide and oseltamivir combinations is more effective at reducing the impact of influenza a virus infection compared to oseltamivir monotherapy","volume":"176","author":"EJ Mifsud","year":"2020","journal-title":"Antiviral Res"},{"issue":"12","key":"ppat.1014604.ref018","doi-asserted-by":"crossref","first-page":"2447","DOI":"10.3390\/v15122447","article-title":"Pre-Clinical Evaluation of the Antiviral Activity of Epigalocatechin-3-Gallate, a Component of Green Tea, against Influenza A(H1N1)pdm Viruses","volume":"15","author":"H Stannard","year":"2023","journal-title":"Viruses"},{"key":"ppat.1014604.ref019","doi-asserted-by":"crossref","first-page":"104851","DOI":"10.1016\/j.antiviral.2020.104851","article-title":"Host-targeted nitazoxanide has a high barrier to resistance but does not reduce the emergence or proliferation of oseltamivir-resistant influenza viruses in vitro or in vivo when used in combination with oseltamivir","volume":"180","author":"D Tilmanis","year":"2020","journal-title":"Antiviral Res"},{"issue":"515","key":"ppat.1014604.ref020","doi-asserted-by":"crossref","DOI":"10.1126\/scitranslmed.aax5866","article-title":"Characterization of orally efficacious influenza drug with high resistance barrier in ferrets and human airway epithelia","volume":"11","author":"M Toots","year":"2019","journal-title":"Sci Transl Med"},{"key":"ppat.1014604.ref021","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.trsl.2019.12.002","article-title":"Quantitative efficacy paradigms of the influenza clinical drug candidate EIDD-2801 in the ferret model","volume":"218","author":"M Toots","year":"2020","journal-title":"Transl Res"},{"issue":"1","key":"ppat.1014604.ref022","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1038\/s44298-025-00126-9","article-title":"Characterization of A(H1N1)pdm09 influenza viruses isolated between 2016 and 2019","volume":"3","author":"L Muawan","year":"2025","journal-title":"Npj Viruses"},{"issue":"1","key":"ppat.1014604.ref023","doi-asserted-by":"crossref","first-page":"1452","DOI":"10.1080\/22221751.2022.2076615","article-title":"Pathogenesis and transmission of human seasonal and swine-origin A(H1) influenza viruses in the ferret model","volume":"11","author":"JA Pulit-Penaloza","year":"2022","journal-title":"Emerg Microbes Infect"},{"key":"ppat.1014604.ref024","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.antiviral.2018.09.007","article-title":"Comparison of early and recent influenza A(H1N1)pdm09 isolates harboring or not the H275Y neuraminidase mutation, in vitro and in animal models","volume":"159","author":"Y Abed","year":"2018","journal-title":"Antiviral Res"},{"issue":"2","key":"ppat.1014604.ref025","doi-asserted-by":"crossref","first-page":"155","DOI":"10.3390\/v12020155","article-title":"Comparison of the Pathogenicity in Mice of A(H1N1)pdm09 Viruses Isolated between 2009 and 2015 in Japan","volume":"12","author":"H Mitake","year":"2020","journal-title":"Viruses"},{"issue":"17","key":"ppat.1014604.ref026","doi-asserted-by":"crossref","first-page":"9361","DOI":"10.1128\/JVI.01214-12","article-title":"Characterization in vitro and in vivo of pandemic (H1N1) 2009 influenza viruses isolated from patients","volume":"86","author":"T Watanabe","year":"2012","journal-title":"J Virol"},{"issue":"22","key":"ppat.1014604.ref027","doi-asserted-by":"crossref","first-page":"11802","DOI":"10.1128\/JVI.01136-10","article-title":"Virulence-associated substitution D222G in the hemagglutinin of 2009 pandemic influenza A(H1N1) virus affects receptor binding","volume":"84","author":"S Chutinimitkul","year":"2010","journal-title":"J Virol"},{"issue":"1","key":"ppat.1014604.ref028","doi-asserted-by":"crossref","DOI":"10.1128\/mSphere.01043-20","article-title":"Next-Generation Sequencing Analysis of the Within-Host Genetic Diversity of Influenza A(H1N1)pdm09 Viruses in the Upper and Lower Respiratory Tracts of Patients with Severe Influenza","volume":"6","author":"I Takayama","year":"2021","journal-title":"mSphere"},{"issue":"1","key":"ppat.1014604.ref029","article-title":"Differential disease severity and whole-genome sequence analysis for human influenza A\/H1N1pdm virus in 2015-2016 influenza season","volume":"7","author":"H Liu","year":"2021","journal-title":"Virus Evol"},{"issue":"7195","key":"ppat.1014604.ref030","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1038\/nature06945","article-title":"The genomic and epidemiological dynamics of human influenza A virus","volume":"453","author":"A Rambaut","year":"2008","journal-title":"Nature"},{"issue":"3","key":"ppat.1014604.ref031","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1016\/j.chom.2013.02.008","article-title":"Defining influenza A virus hemagglutinin antigenic drift by sequential monoclonal antibody selection","volume":"13","author":"SR Das","year":"2013","journal-title":"Cell Host Microbe"},{"key":"ppat.1014604.ref032","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/B978-012220360-2\/50015-5","article-title":"Antigenic Variation in Influenza Viruses","author":"RG Webster","year":"1999","journal-title":"Origin and Evolution of Viruses. Elsevier"},{"issue":"20","key":"ppat.1014604.ref033","doi-asserted-by":"crossref","first-page":"8432","DOI":"10.1073\/pnas.1100768108","article-title":"Influenza virus aerosol exposure and analytical system for ferrets","volume":"108","author":"KM Gustin","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"9","key":"ppat.1014604.ref034","doi-asserted-by":"crossref","first-page":"2458","DOI":"10.1093\/jac\/dku146","article-title":"Evaluation of oseltamivir prophylaxis regimens for reducing influenza virus infection, transmission and disease severity in a ferret model of household contact","volume":"69","author":"DY Oh","year":"2014","journal-title":"J Antimicrob Chemother"},{"issue":"4","key":"ppat.1014604.ref035","article-title":"Baloxavir treatment of ferrets infected with influenza A(H1N1)pdm09 virus reduces onward transmission","volume":"16","author":"LYY Lee","year":"2020","journal-title":"PLoS Pathog"},{"issue":"11","key":"ppat.1014604.ref036","article-title":"Baloxavir Treatment Delays Influenza B Virus Transmission in Ferrets and Results in Limited Generation of Drug-Resistant Variants","volume":"65","author":"PNQ Pascua","year":"2021","journal-title":"Antimicrob Agents Chemother"},{"issue":"16","key":"ppat.1014604.ref037","article-title":"Induction of Interferon-Stimulated Genes Correlates with Reduced Growth of Influenza A Virus in Lungs after RIG-I Agonist Treatment of Ferrets","volume":"96","author":"LSU Schwab","year":"2022","journal-title":"J Virol"},{"issue":"6","key":"ppat.1014604.ref038","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1008592","article-title":"Utilising animal models to evaluate oseltamivir efficacy against influenza A and B viruses with reduced in vitro susceptibility","volume":"16","author":"R Farrukee","year":"2020","journal-title":"PLoS pathogens"},{"issue":"4","key":"ppat.1014604.ref039","doi-asserted-by":"crossref","DOI":"10.1128\/mbio.01174-22","article-title":"Robustness of the Ferret Model for Influenza Risk Assessment Studies: a Cross-Laboratory Exercise","volume":"13","author":"JA Belser","year":"2022","journal-title":"mBio"},{"issue":"15","key":"ppat.1014604.ref040","doi-asserted-by":"crossref","first-page":"7590","DOI":"10.1128\/JVI.01623-05","article-title":"Kinetics of influenza A virus infection in humans","volume":"80","author":"P Baccam","year":"2006","journal-title":"J Virol"},{"issue":"15","key":"ppat.1014604.ref041","doi-asserted-by":"crossref","first-page":"7572","DOI":"10.1128\/JVI.00512-11","article-title":"Comparative pathology in ferrets infected with H1N1 influenza A viruses isolated from different hosts","volume":"85","author":"JH Smith","year":"2011","journal-title":"J Virol"},{"key":"ppat.1014604.ref042","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1186\/1743-422X-7-149","article-title":"Course of seasonal influenza A\/Brisbane\/59\/07 H1N1 infection in the ferret","volume":"7","author":"A McBrayer","year":"2010","journal-title":"Virol J"},{"issue":"4","key":"ppat.1014604.ref043","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0094090","article-title":"Low dose influenza virus challenge in the ferret leads to increased virus shedding and greater sensitivity to oseltamivir","volume":"9","author":"AC Marriott","year":"2014","journal-title":"PLoS One"},{"issue":"3","key":"ppat.1014604.ref044","article-title":"A novel video tracking method to evaluate the effect of influenza infection and antiviral treatment on ferret activity","volume":"10","author":"DY Oh","year":"2015","journal-title":"PLoS One"},{"issue":"1","key":"ppat.1014604.ref045","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1038\/s42003-026-09607-w","article-title":"Oseltamivir and baloxavir monotherapy and combination therapy efficacy against clade 2.3.4.4b A(H5N1) influenza virus infection in ferrets","volume":"9","author":"JA Belser","year":"2026","journal-title":"Commun Biol"},{"issue":"10","key":"ppat.1014604.ref046","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.1716771115","article-title":"Defining the sizes of airborne particles that mediate influenza transmission in ferrets","volume":"115","author":"J Zhou","year":"2018","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"4","key":"ppat.1014604.ref047","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2419985122","article-title":"Dispersal of influenza virus populations within the respiratory tract shapes their evolutionary potential","volume":"122","author":"LM Ferreri","year":"2025","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ppat.1014604.ref048","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/j.virol.2015.06.020","article-title":"Oseltamivir inhibits influenza virus replication and transmission following ocular-only aerosol inoculation of ferrets","volume":"484","author":"JA Belser","year":"2015","journal-title":"Virology"},{"issue":"4","key":"ppat.1014604.ref049","doi-asserted-by":"crossref","first-page":"2226","DOI":"10.1128\/JVI.01565-12","article-title":"Virulence determinants of pandemic A(H1N1)2009 influenza virus in a mouse model","volume":"87","author":"R Uraki","year":"2013","journal-title":"J Virol"},{"issue":"19","key":"ppat.1014604.ref050","doi-asserted-by":"crossref","DOI":"10.1128\/JVI.01206-18","article-title":"Functional Evolution of the 2009 Pandemic H1N1 Influenza Virus NS1 and PA in Humans","volume":"92","author":"A Nogales","year":"2018","journal-title":"J Virol"},{"issue":"5","key":"ppat.1014604.ref051","article-title":"Pathogenicity of the H1N1 influenza virus enhanced by functional synergy between the NPV100I and NAD248N pair","volume":"14","author":"W-J Kim","year":"2019","journal-title":"PLoS One"},{"issue":"22","key":"ppat.1014604.ref052","doi-asserted-by":"crossref","first-page":"13269","DOI":"10.1128\/JVI.01636-14","article-title":"Accumulation of human-adapting mutations during circulation of A(H1N1)pdm09 influenza virus in humans in the United Kingdom","volume":"88","author":"RA Elderfield","year":"2014","journal-title":"J Virol"},{"issue":"6","key":"ppat.1014604.ref053","article-title":"Genomic polymorphism of the pandemic A (H1N1) influenza viruses correlates with viral replication, virulence, and pathogenicity in vitro and in vivo","volume":"6","author":"L Xu","year":"2011","journal-title":"PLoS One"},{"issue":"17","key":"ppat.1014604.ref054","doi-asserted-by":"crossref","DOI":"10.1128\/JVI.00721-17","article-title":"Functional Evolution of Influenza Virus NS1 Protein in Currently Circulating Human 2009 Pandemic H1N1 Viruses","volume":"91","author":"AM Clark","year":"2017","journal-title":"J Virol"},{"issue":"9","key":"ppat.1014604.ref055","doi-asserted-by":"crossref","DOI":"10.1128\/mbio.01017-25","article-title":"The (digestive) path less traveled: influenza A virus and the gastrointestinal tract","volume":"16","author":"TA Bullock","year":"2025","journal-title":"mBio"},{"issue":"3","key":"ppat.1014604.ref056","doi-asserted-by":"crossref","first-page":"800","DOI":"10.3181\/00379727-122-31255","article-title":"Human influenza resulting from aerosol inhalation","volume":"122","author":"RH Alford","year":"1966","journal-title":"Proc Soc Exp Biol Med"},{"key":"ppat.1014604.ref057","doi-asserted-by":"crossref","DOI":"10.1093\/jac\/dki018","article-title":"Oseltamivir (Tamiflu) and its potential for use in the event of an influenza pandemic","volume":"55","author":"P Ward","year":"2005","journal-title":"Journal of Antimicrobial Chemotherapy"},{"issue":"2","key":"ppat.1014604.ref058","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.antiviral.2011.05.008","article-title":"Assessment of the efficacy of the neuraminidase inhibitor oseltamivir against 2009 pandemic H1N1 influenza virus in ferrets","volume":"91","author":"EA Govorkova","year":"2011","journal-title":"Antiviral Res"},{"key":"ppat.1014604.ref059","doi-asserted-by":"crossref","DOI":"10.1093\/infdis\/jiaf263","article-title":"Modeling influenza antiviral strategies: reducing burden and preventing resistance","volume":"232","author":"R Pasco","year":"2025","journal-title":"The Journal of Infectious Diseases"},{"issue":"4","key":"ppat.1014604.ref060","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pcbi.1003568","article-title":"Impact of different oseltamivir regimens on treating influenza A virus infection and resistance emergence: insights from a modelling study","volume":"10","author":"L Canini","year":"2014","journal-title":"PLoS Comput Biol"},{"issue":"7","key":"ppat.1014604.ref061","doi-asserted-by":"crossref","first-page":"101070","DOI":"10.1016\/j.lanmic.2024.101070","article-title":"Ocular infectivity and replication of a clade 2.3.4.4b A(H5N1) influenza virus associated with human conjunctivitis in a dairy farm worker in the USA: an in-vitro and ferret study","volume":"6","author":"JA Belser","year":"2025","journal-title":"Lancet Microbe"},{"issue":"1","key":"ppat.1014604.ref062","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1038\/s42003-022-04005-4","article-title":"Assessing the fitness of a dual-antiviral drug resistant human influenza virus in the ferret model","volume":"5","author":"HL Stannard","year":"2022","journal-title":"Commun Biol"},{"issue":"14","key":"ppat.1014604.ref063","doi-asserted-by":"crossref","first-page":"2490","DOI":"10.1093\/bioinformatics\/bty121","article-title":"Parallelization of MAFFT for large-scale multiple sequence alignments","volume":"34","author":"T Nakamura","year":"2018","journal-title":"Bioinformatics"},{"issue":"5","key":"ppat.1014604.ref064","doi-asserted-by":"crossref","first-page":"1530","DOI":"10.1093\/molbev\/msaa015","article-title":"IQ-TREE 2: New Models and Efficient Methods for Phylogenetic Inference in the Genomic Era","volume":"37","author":"BQ Minh","year":"2020","journal-title":"Mol Biol Evol"},{"issue":"1","key":"ppat.1014604.ref065","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1093\/sysbio\/syv068","article-title":"Fast Dating Using Least-Squares Criteria and Algorithms","volume":"65","author":"T-H To","year":"2016","journal-title":"Syst Biol"},{"issue":"1","key":"ppat.1014604.ref066","article-title":"Using ggtree to Visualize Data on Tree-Like Structures","volume":"69","author":"G Yu","year":"2020","journal-title":"Curr Protoc Bioinformatics"},{"issue":"67","key":"ppat.1014604.ref067","article-title":"Nextclade: clade assignment, mutation calling and quality control for viral genomes","volume":"6","author":"IRC Aksamentov","year":"2021","journal-title":"Journal of Open Source Software"},{"issue":"3","key":"ppat.1014604.ref068","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A Simple Method Of Estimating Fifty Per Cent Endpoints12","volume":"27","author":"Lj Reed","year":"1938","journal-title":"American Journal of Epidemiology"},{"issue":"6","key":"ppat.1014604.ref069","first-page":"900","article-title":"Kinetics of lung lesion development and pro-inflammatory cytokine response in pigs with vaccine-associated enhanced respiratory disease induced by challenge with pandemic (2009) A\/H1N1 influenza virus","volume":"49","author":"PC Gauger","year":"2012","journal-title":"Veterinary Microbiology"},{"issue":"2","key":"ppat.1014604.ref070","doi-asserted-by":"crossref","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":"S Hussain","year":"2019","journal-title":"J Virol"},{"issue":"12","key":"ppat.1014604.ref071","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1002443","article-title":"Eurasian-origin gene segments contribute to the transmissibility, aerosol release, and morphology of the 2009 pandemic H1N1 influenza virus","volume":"7","author":"SS Lakdawala","year":"2011","journal-title":"PLoS Pathog"},{"issue":"4","key":"ppat.1014604.ref072","doi-asserted-by":"crossref","DOI":"10.1128\/aem.02271-21","article-title":"Inherent heterogeneity of influenza A virus stability following aerosolization","volume":"88","author":"JA Belser","year":"2022","journal-title":"Applied and Environmental Microbiology"},{"issue":"3","key":"ppat.1014604.ref073","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1002569","article-title":"Influenza virus respiratory infection and transmission following ocular inoculation in ferrets","volume":"8","author":"JA Belser","year":"2012","journal-title":"PLoS Pathog"},{"issue":"19","key":"ppat.1014604.ref074","doi-asserted-by":"crossref","first-page":"10309","DOI":"10.1128\/JVI.01109-09","article-title":"Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and Swine origin human influenza a viruses","volume":"83","author":"B Zhou","year":"2009","journal-title":"J Virol"}],"updated-by":[{"DOI":"10.1371\/journal.ppat.1014604","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000}}],"container-title":["PLOS Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1014604","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T18:05:51Z","timestamp":1789754751000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1014604"}},"subtitle":[],"editor":[{"given":"Mark T.","family":"Heise","sequence":"first","affiliation":[{"name":"University of North Carolina at Chapel Hill","place":["UNITED STATES OF AMERICA"]}],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,9]]},"references-count":74,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9,9]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.ppat.1014604","relation":{},"ISSN":["1553-7374"],"issn-type":[{"value":"1553-7374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,9]]}}}