Equine Influenza Culture Methods

  • Thomas M. ChambersEmail author
  • Stephanie E. Reedy
Part of the Methods in Molecular Biology book series (MIMB, volume 2123)


Equine influenza viruses are cultured in embryonated chicken eggs or in mammalian cells, generally Madin-Darby canine kidney (MDCK) cells, using methods much the same as for other influenza A viruses. Mutations associated with host adaptation occur in both eggs and MDCK cells, but the latter show greater heterogeneity and eggs are the generally preferred host. Both equine-1 H7N7 and equine-2 H3N8 viruses replicate efficiently in 11-day-old eggs, but we find that equine-1 viruses kill the embryos whereas equine-2 viruses do not.

Key words

Embryonated eggs Mammalian influenza MDCK Plaque assay TCID assay 


  1. 1.
    Beveridge WIB, Burnet FM (1946) In: Council MR (ed) The cultivation of viruses and rickettsiae in the chick embryo. His Majesty’s Stationery Office, LondonGoogle Scholar
  2. 2.
    Merten OW, Hannoun C, Manuguerra JC, Ventre F, Petres S (1996) Production of influenza virus in cell cultures for vaccine preparation. Adv Exp Med Biol 397:141–151CrossRefGoogle Scholar
  3. 3.
    Quinlivan M, Zamarin D, Garcia-Sastre A, Cullinane A, Chambers T, Palese P (2005) Attenuation of equine influenza viruses through truncations of the NS1 protein. J Virol 79:8431–8439CrossRefGoogle Scholar
  4. 4.
    Yamagishi H, Ide S, Eiki T, Eiguchi Y, Nagamine T, Igarashi Y, Yoshioka I, Matumoto M (1984) Plaque assay of equine influenza virus. Vet Microbiol 9:187–192CrossRefGoogle Scholar
  5. 5.
    Ilobi CP, Henfrey R, Robertson JS, Mumford JA, Erasmus BJ, Wood JM (1994) Antigenic and molecular characterization of host cell-mediated variants of equine H3N8 influenza viruses. J Gen Virol 75(Pt 3):669–673CrossRefGoogle Scholar
  6. 6.
    Ilobi CP, Nicolson C, Taylor J, Mumford JA, Wood JM, Robertson JS (1998) Direct sequencing of the HA gene of clinical equine H3N8 influenza virus and comparison with laboratory derived viruses. Arch Virol 143:891–901CrossRefGoogle Scholar
  7. 7.
    Cook RF, Cook SJ (1991) Differences in sensitivity in haemagglutinin inhibition assays between A/equine/H3N8 viruses isolated in eggs and MDCK cells are linked to cleavage of the haemagglutinin molecule. Vet Microbiol 27:253–261CrossRefGoogle Scholar
  8. 8.
    Cook RF, Mumford JA, Douglas A, Wood JM (1988) The influence of the host cell on the antigenic properties of equine-2 influenza A viruses. In: Equine Infectious Diseases V: Proceedings of the Fifth International Conference. Powell DG (ed), University Press of Kentucky, Lexington, pp. 60–65Google Scholar
  9. 9.
    OIE (2012) Manual of diagnostic tests and vaccines for terrestrial animals, 7th edn. World Organisation for Animal Health (OIE), Paris, pp 865–878. Ch 2.5.7Google Scholar
  10. 10.
    Zhang L (2011) Differentiated innate immune responses correlate with the contrasting pathogenicity of the equine H7N7 influenza demonstrated in horses and Balb/C mice. University of Kentucky, Lexington, KYGoogle Scholar

Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply. 2020

Authors and Affiliations

  1. 1.Maxwell H. Gluck Equine Research Center, Department of Veterinary ScienceUniversity of KentuckyLexingtonUSA

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