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Viruses2022; 14(4); doi: 10.3390/v14040661

Experimental Infection of Horses with Influenza D Virus.

Abstract: Antibodies to influenza D virus (IDV) have been detected in horses, but no evidence of disease in the field has been reported. To determine whether IDV is infectious, immunogenic, and pathogenic in horses, four 2-year-old horses seronegative for both influenza A (H3N8) and D viruses were intranasally inoculated with 6.25 × 107 TCID50/animal of D/bovine/California/0363/2019 (D/CA2019) virus, using a portable equine nebulizer system. Horses were observed daily for clinical signs including rectal temperature, nasal discharge, coughing, lung sounds, tachycardia, and tachypnea. No horses exhibited clinical signs of disease. Nasopharyngeal swabs collected from 1-8 days post-infection demonstrated virus shedding by qRT-PCR. The horses showed evidence of seroconversion as early as 13 days post-infection (dpi) and the geometric mean of the antibody titers (GMT) of all four horses ranged from 16.82-160 as demonstrated by the microneutralization assay. Further, deep RNA sequencing of the virus isolated in embryonated chicken eggs revealed no adaptive mutations indicating that IDV can replicate in horses, suggesting the possibility of interspecies transmission of IDV with bovine reservoir into equids in nature.
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Publication Date: 2022-03-23 PubMed ID: 35458390PubMed Central: PMC9029652DOI: 10.3390/v14040661Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research explored if the influenza D virus (IDV) can infect, trigger an immune response, and cause diseases in horses. Using four 2-year-old horses, the experiment found that while the horses didn’t exhibit any disease symptoms, they shed the virus and developed antibodies against it. Sequencing of the isolated virus showed no mutations, indicating that IDV can replicate in horses, potentially leading to interspecies transmission between cattle and horses.

Research Methodology

  • Four 2-year-old horses that had not been exposed to influenza A or D were selected for the study.
  • Each horse was inoculated with the D/bovine/California/0363/2019 (D/CA2019) strain of IDV using a portable equine nebulizer system.
  • The horses’ health was monitored daily for signs of illness such as fever, nasal discharge, cough, abnormal lung sounds, and rapid heart rate or breathing.
  • Nasopharyngeal swabs were taken from the horses from day 1 to day 8 post-infection to check for virus shedding.

Results

  • The horses did not exhibit any clinical signs of illness.
  • The nasopharyngeal swabs demonstrated that the horses were shedding the virus, as detected by the qRT PCR test.
  • Seroconversion, or the development of detectable antibodies in the blood against the virus, was observed as early as 13 days post-infection.
  • The geometric mean of antibody titers of all horses ranged from 16.82-160, measured by a microneutralization assay.
  • Sequencing of the virus isolated in embryonated chicken eggs showed no adaptive mutations, indicating that IDV can replicate in horses without changing its genetic composition.

Implications

  • The findings suggest that IDV is infectious and immunogenic in horses, as shown by the virus shedding and the development of antibodies.
  • However, it doesn’t necessarily cause disease or visible symptoms in horses, as none of the studied individuals showed clinical signs of infection.
  • The ability to replicate in horses without any adaptive mutations points to the risk of interspecies transmission of IDV, particularly from cattle, a known reservoir, to horses.

Cite This Article

APA
Sreenivasan CC, Uprety T, Reedy SE, Temeeyasen G, Hause BM, Wang D, Li F, Chambers TM. (2022). Experimental Infection of Horses with Influenza D Virus. Viruses, 14(4). https://doi.org/10.3390/v14040661

Publication

Viruses
ISSN: 1999-4915
NlmUniqueID: 101509722
Country: Switzerland
Language: English
Volume: 14
Issue: 4

Researcher Affiliations

Sreenivasan, Chithra C
  • Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Uprety, Tirth
  • Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Reedy, Stephanie E
  • Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Temeeyasen, Gun
  • Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, USA.
Hause, Ben M
  • Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, USA.
Wang, Dan
  • Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Li, Feng
  • Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Chambers, Thomas M
  • Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.

MeSH Terms

  • Animals
  • Antibodies, Viral
  • Cattle
  • Horse Diseases
  • Horses
  • Influenza A Virus, H3N8 Subtype
  • Orthomyxoviridae
  • Orthomyxoviridae Infections
  • Thogotovirus

Grant Funding

  • R01 AI141889 / NIAID NIH HHS

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 31 references
  1. Dionisio L, Medeiros F, Pequito M, Faustino-Rocha AI. Equine influenza: A comprehensive review from etiology to treatment.. Anim. Health Res. Rev. 2021;22:56–71.
    doi: 10.1017/S1466252321000050pubmed: 34075870google scholar: lookup
  2. Nedland H, Wollman J, Sreenivasan C, Quast M, Singrey A, Fawcett L, Christopher-Hennings J, Nelson E, Kaushik RS, Wang D. Serological evidence for the co-circulation of two lineages of influenza D viruses in equine populations of the Midwest United States.. Zoonoses Public Health 2018;65:e148–e154.
    doi: 10.1111/zph.12423pmc: PMC5766371pubmed: 29139222google scholar: lookup
  3. Quast M, Sreenivasan C, Sexton G, Nedland H, Singrey A, Fawcett L, Miller G, Lauer D, Voss S, Pollock S. Serological evidence for the presence of influenza D virus in small ruminants.. Vet. Microbiol. 2015;180:281–285.
    doi: 10.1016/j.vetmic.2015.09.005pmc: PMC4618254pubmed: 26414999google scholar: lookup
  4. Salem E, Cook EAJ, Lbacha HA, Oliva J, Awoume F, Aplogan GL, Hymann EC, Muloi D, Deem SL, Alali S. Serologic Evidence for Influenza C and D Virus among Ruminants and Camelids, Africa, 1991–2015.. Emerg. Infect. Dis. 2017;23:1556–1559.
    doi: 10.3201/eid2309.170342pmc: PMC5572875pubmed: 28820371google scholar: lookup
  5. Zhai SL, Zhang H, Chen SN, Zhou X, Lin T, Liu R, Lv DH, Wen XH, Wei WK, Wang D. Influenza D Virus in Animal Species in Guangdong Province, Southern China.. Emerg. Infect. Dis. 2017;23:1392–1396.
    doi: 10.3201/eid2308.170059pmc: PMC5547803pubmed: 28726609google scholar: lookup
  6. Trombetta CM, Montomoli E, Di Bartolo I, Ostanello F, Chiapponi C, Marchi S. Detection of antibodies against influenza D virus in swine veterinarians in Italy in 2004.. J. Med. Virol. 2021.
    doi: 10.1002/jmv.27466pmc: PMC9299042pubmed: 34811769google scholar: lookup
  7. Trombetta CM, Marchi S, Manini I, Kistner O, Li F, Piu P, Manenti A, Biuso F, Sreenivasan C, Druce J. Influenza D Virus: Serological Evidence in the Italian Population from 2005 to 2017.. Viruses 2019;12:30.
    doi: 10.3390/v12010030pmc: PMC7019439pubmed: 31892120google scholar: lookup
  8. White SK, Ma W, McDaniel CJ, Gray GC, Lednicky JA. Serologic evidence of exposure to influenza D virus among persons with occupational contact with cattle.. J. Clin. Virol. 2016;81:31–33.
    doi: 10.1016/j.jcv.2016.05.017pubmed: 27294672google scholar: lookup
  9. Daly JM, Wu G, Norman F, Preston T, Kydd JH, Jengarn J, Scott SD, Temperton N, Rash A, Elton DM. Limited evidence for exposure of UK horses to influenza D virus.. Equine Vet. J. 2021;53:81.
    doi: 10.1111/evj.127_13495google scholar: lookup
  10. Bailey ES, Fieldhouse JK, Alarja NA, Chen DD, Kovalik ME, Zemke JN, Choi JY, Borkenhagen LK, Toh TH, Lee JSY. First sequence of influenza D virus identified in poultry farm bioaerosols in Sarawak, Malaysia.. Trop. Dis. Travel Med. Vaccines 2020;6:5.
    doi: 10.1186/s40794-020-0105-9pmc: PMC7069008pubmed: 32190346google scholar: lookup
  11. Borkenhagen LK, Mallinson KA, Tsao RW, Ha SJ, Lim WH, Toh TH, Anderson BD, Fieldhouse JK, Philo SE, Chong KS. Surveillance for respiratory and diarrheal pathogens at the human-pig interface in Sarawak, Malaysia.. PLoS ONE 2018;13:e0201295.
    doi: 10.1371/journal.pone.0201295pmc: PMC6063427pubmed: 30052648google scholar: lookup
  12. Bailey ES, Choi JY, Zemke J, Yondon M, Gray GC. Molecular surveillance of respiratory viruses with bioaerosol sampling in an airport.. Trop. Dis. Travel Med. Vaccines 2018;4:11.
    doi: 10.1186/s40794-018-0071-7pmc: PMC6142699pubmed: 30237898google scholar: lookup
  13. Anderson BD, Yondon M, Bailey ES, Duman EK, Simmons RA, Greer AG, Gray GC. Environmental bioaerosol surveillance as an early warning system for pathogen detection in North Carolina swine farms: A pilot study.. Transbound Emerg. Dis. 2021;68:361–367.
    doi: 10.1111/tbed.13683pubmed: 32535997google scholar: lookup
  14. Choi JY, Zemke J, Philo SE, Bailey ES, Yondon M, Gray GC. Aerosol Sampling in a Hospital Emergency Room Setting: A Complementary Surveillance Method for the Detection of Respiratory Viruses.. Front. Public Health 2018;6:174.
    doi: 10.3389/fpubh.2018.00174pmc: PMC6011129pubmed: 29963543google scholar: lookup
  15. Barnard KN, Alford-Lawrence BK, Buchholz DW, Wasik BR, LaClair JR, Yu H, Honce R, Ruhl S, Pajic P, Daugherity EK. Modified Sialic Acids on Mucus and Erythrocytes Inhibit Influenza A Virus Hemagglutinin and Neuraminidase Functions.. J. Virol. 2020;94.
    doi: 10.1128/JVI.01567-19pmc: PMC7163148pubmed: 32051275google scholar: lookup
  16. Nemanichvili N, Berends AJ, Wubbolts RW, Grone A, Rijks JM, de Vries RP, Verheije MH. Tissue Microarrays to Visualize Influenza D Attachment to Host Receptors in the Respiratory Tract of Farm Animals.. Viruses 2021;13:586.
    doi: 10.3390/v13040586pmc: PMC8067312pubmed: 33807137google scholar: lookup
  17. Mazzetto E, Bortolami A, Fusaro A, Mazzacan E, Maniero S, Vascellari M, Beato MS, Schiavon E, Chiapponi C, Terregino C. Replication of Influenza D Viruses of Bovine and Swine Origin in Ovine Respiratory Explants and Their Attachment to the Respiratory Tract of Bovine, Sheep, Goat, Horse, and Swine.. Front. Microbiol. 2020;11:1136.
    doi: 10.3389/fmicb.2020.01136pmc: PMC7261881pubmed: 32523585google scholar: lookup
  18. Reed LJ, Muench H. A simple method of estimating fifty percent endpoints.. Am. J. Epidemiol. 1938;27:493–497.
    doi: 10.1093/oxfordjournals.aje.a118408google scholar: lookup
  19. Blanco-Lobo P, Rodriguez L, Reedy S, Oladunni FS, Nogales A, Murcia PR, Chambers TM, Martinez-Sobrido L. A Bivalent Live-Attenuated Vaccine for the Prevention of Equine Influenza Virus.. Viruses 2019;11:933.
    doi: 10.3390/v11100933pmc: PMC6832603pubmed: 31614538google scholar: lookup
  20. Chambers TM, Reedy SE. Equine Influenza Culture Methods.. Methods Mol. Biol. 2020;2123:393–400.
    doi: 10.1007/978-1-0716-0346-8_30pubmed: 32170705google scholar: lookup
  21. Hause BM, Ducatez M, Collin EA, Ran Z, Liu R, Sheng Z, Armien A, Kaplan B, Chakravarty S, Hoppe AD. Isolation of a novel swine influenza virus from Oklahoma in 2011 which is distantly related to human influenza C viruses.. PLoS Pathog. 2013;9:e1003176.
    doi: 10.1371/journal.ppat.1003176pmc: PMC3567177pubmed: 23408893google scholar: lookup
  22. Hause BM, Nelson EA, Christopher-Hennings J, Pipas JM. North American Big Brown Bats (Eptesicus fuscus) Harbor an Exogenous Deltaretrovirus.. mSphere 2020;5:e00902.
    doi: 10.1128/mSphere.00902-20pmc: PMC7568648pubmed: 32968009google scholar: lookup
  23. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool.. J. Mol. Biol. 1990;215:403–410.
    doi: 10.1016/S0022-2836(05)80360-2pubmed: 2231712google scholar: lookup
  24. . Manual for the Laboratory Diagnosis and Virological Surveillance of Influenza.. World Health Organization 2011.
  25. Wasik BR, Barnard KN, Ossiboff RJ, Khedri Z, Feng KH, Yu H, Chen X, Perez DR, Varki A, Parrish CR. Distribution of O-Acetylated Sialic Acids among Target Host Tissues for Influenza Virus.. mSphere 2017;2:e00379-16.
    doi: 10.1128/mSphere.00379-16pmc: PMC5588038pubmed: 28904995google scholar: lookup
  26. Huang C, Yu J, Hause BM, Park JY, Sreenivasan C, Uprety T, Sheng Z, Wang D, Li F. Emergence of new phylogenetic lineage of Influenza D virus with broad antigenicity in California, United States.. Emerg. Microbes Infect. 2021;10:739–742.
    doi: 10.1080/22221751.2021.1910078pmc: PMC8043534pubmed: 33771071google scholar: lookup
  27. Sreenivasan CC, Sheng Z, Wang D, Li F. Host Range, Biology, and Species Specificity of Seven-Segmented Influenza Viruses—A Comparative Review on Influenza C and D.. Pathogens 2021;10:1583.
    doi: 10.3390/pathogens10121583pmc: PMC8704295pubmed: 34959538google scholar: lookup
  28. Ferguson L, Olivier AK, Genova S, Epperson WB, Smith DR, Schneider L, Barton K, McCuan K, Webby RJ, Wan XF. Pathogenesis of Influenza D Virus in Cattle.. J. Virol. 2016;90:5636–5642.
    doi: 10.1128/JVI.03122-15pmc: PMC4886773pubmed: 27030270google scholar: lookup
  29. Salem E, Hagglund S, Cassard H, Corre T, Naslund K, Foret C, Gauthier D, Pinard A, Delverdier M, Zohari S. Pathogenesis, Host Innate Immune Response, and Aerosol Transmission of Influenza D Virus in Cattle.. J. Virol. 2019;93:e01853-18.
    doi: 10.1128/JVI.01853-18pmc: PMC6430558pubmed: 30674628google scholar: lookup
  30. Ferguson L, Luo K, Olivier AK, Cunningham FL, Blackmon S, Hanson-Dorr K, Sun H, Baroch J, Lutman MW, Quade B. Influenza D Virus Infection in Feral Swine Populations, United States.. Emerg. Infect. Dis. 2018;24:1020–1028.
    doi: 10.3201/eid2406.172102pmc: PMC6004836pubmed: 29774857google scholar: lookup
  31. Kaplan BS, Falkenberg S, Dassanayake R, Neill J, Velayudhan B, Li F, Vincent AL. Virus strain influenced the interspecies transmission of influenza D virus between calves and pigs.. Transbound Emerg. Dis. 2021;68:3396–3404.
    doi: 10.1111/tbed.13943pubmed: 33259672google scholar: lookup

Citations

This article has been cited 8 times.
  1. Li H, Yan W, Liu X, Gao B, Peng J, Jiang F, Cui Q, Song C, Kong X, Li H, Stoeger T, Wajid A, Dodovski A, Gao C, Lusida MI, Mingala CN, Andreychuk DB, Yin R. Emergence and Phylodynamics of Influenza D Virus in Northeast China Reveal Sporadic Detection and Predominance of the D/Yamagata/2019 Lineage in Cattle. Viruses 2026 Jan 9;18(1).
    doi: 10.3390/v18010093pubmed: 41600857google scholar: lookup
  2. Falsini A, Coppola C, Fiori A, Buonavoglia D, Marchi S, Montomoli E, Pellegrini F, Lanave G, Martella V, Camero M, Trombetta CM. Influenza D Virus Circulation Among Bovines, Swine, Equines, and Wild Boars in Italy: A Sero-Epidemiological Study. Pathogens 2025 Sep 5;14(9).
    doi: 10.3390/pathogens14090891pubmed: 41011791google scholar: lookup
  3. Umar S, Ahmed A, Gulraiz SH, Muhammad S, Yu J, Rasool A, Koviazina R, Yilmaz A, Yilmaz H, Anderson BD. First Report of Influenza D Virus in Dairy Cattle in Pakistan. Viruses 2024 Nov 29;16(12).
    doi: 10.3390/v16121865pubmed: 39772175google scholar: lookup
  4. Trombetta CM, Marchi S, Marotta MG, Moreno A, Chiapponi C, Montomoli E, Lanave G, Camero M, Martella V. Detection of Influenza D Antibodies in Dogs, Apulia Region, Italy, 2016 and 2023. Emerg Infect Dis 2024 May;30(5):1045-1047.
    doi: 10.3201/eid3005.231401pubmed: 38666735google scholar: lookup
  5. Kwasnik M, Rola J, Rozek W. Influenza D in Domestic and Wild Animals. Viruses 2023 Dec 15;15(12).
    doi: 10.3390/v15122433pubmed: 38140674google scholar: lookup
  6. Uprety T, Sreenivasan CC, Thomas M, Hause B, Christopher-Hennings J, Miskimis D, Pillatzki A, Nelson E, Wang D, Li F. Prevalence and characterization of seven-segmented influenza viruses in bovine respiratory disease complex. Virology 2023 Oct;587:109859.
    doi: 10.1016/j.virol.2023.109859pubmed: 37544044google scholar: lookup
  7. Ruiz M, Puig A, Bassols M, Fraile L, Armengol R. Influenza D Virus: A Review and Update of Its Role in Bovine Respiratory Syndrome. Viruses 2022 Dec 5;14(12).
    doi: 10.3390/v14122717pubmed: 36560721google scholar: lookup
  8. Gaudino M, Chiapponi C, Moreno A, Zohari S, O'Donovan T, Quinless E, Sausy A, Oliva J, Salem E, Fusade-Boyer M, Meyer G, Hübschen JM, Saegerman C, Ducatez MF, Snoeck CJ. Evolutionary and temporal dynamics of emerging influenza D virus in Europe (2009-22). Virus Evol 2022;8(2):veac081.
    doi: 10.1093/ve/veac081pubmed: 36533151google scholar: lookup
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