Journal of tissue culture methods

, Volume 11, Issue 3, pp 139–145 | Cite as

Cell culture propagation of bovine coronavirus

  • Linda J. Saif
  • Robert A. Heckert
  • Kathy L. Miller
  • Mohamed M. Tarek

Summary

Although most field strains of bovine coronavirus (BCV) grow poorly in cell culture and fail to produce cytopathic effects (CPE) until after blind passage, primary calf kidney (PCK) and Vero cells have permitted primary isolation of virus. Cell culture-adapted strains of BCV replicate in PCK, bovine embryonic lung, bovine fetal thyroid, bovine fetal brain, bovine skin cells, ovine fetal kidney cells, and the cell lines pig kidney K3 and 15, Vero, human embryonic lung fibroblasts, HRT-18, MDBK and BEK-1, with trypsin useful for enhancing replication. Organ culture as well as suckling mouse, rat, and hamster brains also support the growth of cell culture-adapted BCV strains. Viral growth is most commonly detected by CPE, immunofluorescence, hemagglutination, and hemadsorption assays or electron microscopy of supernatants from infected cells. In this report, the optimal conditions for the growth and plaque assay of the NCDV strain of BCV in MDBK cells are described.

Key words

bovine coronavirus cell culture propagation 

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References

  1. 1.
    Akashi, H.; Inaba, Y.; Miura, Y., et al. Properties of a coronavirus isolated from a cow with epizootic diarrhea. Vet. Microbiol. 5:265–276; 1980.Google Scholar
  2. 2.
    Akashi, H.; Inaba, Y.; Miura, Y., et al. Propagation of the Kakegawa strain of bovine coronavirus in suckling mice, rats and hamsters. Arch. Virol. 67:367–370; 1981.PubMedGoogle Scholar
  3. 3.
    Barboi, G. H.; Pirvulescu, M. Le test d'inhibition de l'hemagglutination (IHA) pour le diagnostic de l'infection a coronavirus chez les bovins. Rev. Roum. Med.-Virol. 37:253–256; 1986.Google Scholar
  4. 4.
    Baron, S.; McKerlie, M. L. Broadly active inhibitor of viruses spontaneously produced by many cell types in culture. Infect. Immun. 32:449–453; 1981.PubMedGoogle Scholar
  5. 5.
    Bosgiraud, C.; Nicolas, J. A. Pneumonie a coronavirus chez les bovine: A propos de deux observations. Rec. Med. Vet. 162:1085–1086; 1986.Google Scholar
  6. 6.
    Bridger, J. C.; Caul, E. O.; Egglestone, S. I. Replication of an enteric bovine coronavirus in intestinal organ cultures. Arch Virol. 57:43–51; 1978.PubMedGoogle Scholar
  7. 7.
    Cyr-Coats, K. St.; Storz, J. Bovine coronavirus cytopathic expression and plaque formation: host cell and virus strain determine trypsin dependence. 68th Annual Conference of Research Workers in Animal Disease; 1987, November 16–17, Chicago IL., Abstract 83.Google Scholar
  8. 8.
    Dea, S.; Roy, R. S.; Begin, M. E. Bovine coronavirus isolation and cultivation in continuous cell lines. Am. J. Vet. Res. 41:30–38; 1980.PubMedGoogle Scholar
  9. 9.
    Debiaggi, M.; Perduca, M.; Romero, E., et al. Phosphatidyl-serine inhibition of OC43 and NCDCV coronavirus infectivity. Microbiologica 8:313–317; 1985.PubMedGoogle Scholar
  10. 10.
    Deregt, D.; Crouch, C. F.; Sabara, M., et al. Preliminary studies of a bovine coronavirus (BCV) antigen responsible for neutralization. Proceeding, Fourth International Symposium on Neonatal Diarrhea, October 3–5, Saskatoon, Saskatchewan, Canada, Veterinary Infectious Disease Organization, 1984:117–132.Google Scholar
  11. 11.
    Doughri, A. M.; Storz, J.; Hajer, I., et al. Morphology and morphogenesis of a coronavirus infecting intestinal epithelial cells of newborn calves. Exp. Mol. Pathol. 25:355–370; 1976.PubMedGoogle Scholar
  12. 12.
    Gerna, G.; Cereda, P. M.; Revello, M. G., et al. Antigenic and biological relationships between human coronavirus OC43 and neonatal calf diarrhoea coronavirus. J. Gen. Virol. 54:91–102; 1981.PubMedGoogle Scholar
  13. 13.
    Hirano, N.; Sada, Y.; Tuchiya, K., et al. Plaque assay of bovine coronavirus in BEK-1 cells. Jpn. J. Vet. Sci. 47:679–681; 1985.Google Scholar
  14. 14.
    Hughes T. K.; Blalock J. E.; McKerlie M. L., et al. Cell-produced viral inhibitor: possible mechanism of action and chemical composition. Infect. Immun. 32:454–457; 1981.PubMedGoogle Scholar
  15. 15.
    Inaba, Y.; Sato, K.; Kurogi, H., et al. Replication of bovine coronavirus in cell line BEK-1 culture. Arch. Virol. 50:339–342; 1976.PubMedGoogle Scholar
  16. 16.
    Kaye, H. S.; Yarbrough, W. B.; Reed, C. J. Calf diarrhoea coronavirus. Lancet II:509; 1975.Google Scholar
  17. 17.
    L'Haridon, R.; Scherrer, R.; Vautherot, J. F., et al. Adaptation d'un isolement de coronavirus enterique bovine a la culture cellulaire et caracterisation de la souche obtenue. Ann. Rech. Vet. 12:243–251; 1981.PubMedGoogle Scholar
  18. 18.
    Langpap, T. J.; Bergeland, M. E.; Reed, D. E. Coronaviral enteritis of young calves: virologic and pathologic findings in naturally occurring infections. Am. J. Vet. Res. 40:1476–1478; 1979.PubMedGoogle Scholar
  19. 19.
    Laporte, J.; L'Haridon, R.; Bobulesco, P. In vitro culture of bovine enteritic coronavirus (BEC). In: Bricout F.; Pensaert M.; Flewett T. H., et al., compilers. Enterites virales chex l'homme et l'animal; viral Enteritis in humans and animals. Thiverval-Grignon, France: I'INSERM 1979; 90:99–102.Google Scholar
  20. 20.
    Laporte, J.; Bobulesco, P.; Rossi, F. Une lignee cellulaire particulierement sensible a la replication du coronavirus enteritique bovin: les cellules HRT 18. C. R. Acad. Sci. 290:623–626, Serie D; 1980.Google Scholar
  21. 21.
    McNulty, M. S.; Bryson, D. G.; Allan, G. M., et al. Coronavirus infection of the bovine respiratory tract. Vet. Microbiol. 9:425–434; 1984.PubMedGoogle Scholar
  22. 22.
    Mebus, C. A.; Stair, E. L.; Rhodes, M. B., et al. Neonatal calf diarrhea: propagation, attenuation, and characteristics of a coronavirus-like agent. Am. J. Vet. Res. 34:145–150; 1973.PubMedGoogle Scholar
  23. 23.
    Moussa, A. Incurved disc from particles associated with a bovine coronavirus growing in Vero and calf testicle cell cultures. Proceeding, VII International Congress of Virology, August 9–14, Edmonton, Alberta Canada, Abstract R11.11, 1987:110.Google Scholar
  24. 24.
    Phillips, R. W.; Case, G. L. Altered metabolism, acute shock, and therapeutic response in a calf with severe coronavirus-induced diarrhea. Am. J. Vet. Res. 41:1039–1044; 1980.PubMedGoogle Scholar
  25. 25.
    Reynolds, D. J. Coronavirus replication in the intestinal and respiratory tracts during infection of calves. Ann. Rech. Vet. 14:445–446; 1983.PubMedGoogle Scholar
  26. 26.
    Saif, L. J.; Redman, D. R.; Moorhead, P. D., et al. Experimentally induced coronavirus infections in calves: viral replication in the respiratory and intestinal tracts. Am. J. Vet. Res. 47:1426–1432; 1986.PubMedGoogle Scholar
  27. 27.
    Sato, K.; Inaba, Y.; Kurogi, H., et al. Physico-chemical properties of calf diarrhea coronavirus. Vet. Microbiol. 2:73–81; 1977.Google Scholar
  28. 28.
    Sharpee, R. L.; Mebus, C. A.; Bass, E. P. Characterization of a calf diarrheal coronavirus. Am. J. Vet. Res. 37:1031–1041; 1976.PubMedGoogle Scholar
  29. 29.
    Siddell, S.; Wege, H.; Ter Melullen, V. The biology of coronaviruses. J. Gen. Virol. 64:761–776; 1983.PubMedGoogle Scholar
  30. 30.
    Stair, E. L.; Rhodes, M. S.; White, R. G., et al. Neonatal calf diarrhea: purification and electron microscopy of a coronavirus-like agent. Am. J. Vet. Res. 33:1147–1156; 1972.PubMedGoogle Scholar
  31. 31.
    Storz, J.; Rott, R.; Kaluza, G. Enhancement of plaque formation and cell fusion of an enteropathogenic coronavirus by trypsin treatment. Infect. Immun. 31:1214–1222; 1981.PubMedGoogle Scholar
  32. 32.
    Stott, E. J.; Thomas, L. H.; Bridger, J. C., et al. Replication of a bovine coronavirus in organ cultures of foetal trachea. Vet. Microbiol. 1:65–69; 1976.Google Scholar
  33. 33.
    Sturman, L. S.; Homes, K. V. The molecular biology of coronaviruses. Adv. Virus Res. 28:35–112; 1983.PubMedGoogle Scholar
  34. 34.
    Takahashi, E.; Inaba, Y.; Sato, K., et al. Epizootic diarrhoea of adult cattle associated with a coronavirus-like agent. Vet. Microbiol. 5:151–154; 1980.Google Scholar
  35. 35.
    Tektoff, J.; Cauvergne, M.; Durafour, M., et al. Propagation of bovine coronavirus on Vero cell line: electron microscopic studies. In: International Symposium on Enteric Infections in Man and Animals: Standardization of Immunological Procedures. Dublin Ireland Develop. Biol. Standard. 53:299–310; 1982. Basel; S. Karger; 1983.Google Scholar
  36. 36.
    Tektoff, J.; Dauvergne, M.; Durafour, M., et al. Propagation of bovine enteritic coronavirus in three cell systems: electron microscopic studies. Proceeding of the Fourth International Symposium on Neonatal Diarrhea. Veterinary Infectious Disease Organization; 1984:134–153.Google Scholar
  37. 37.
    Thomas, L. H.; Gourlay, R. N.; Stott, E. J., et al. A search for new microorganisms in calf pneumonia by the inoculation of gnotobiotic calves. Res. Vet. Sci. 33:170–182; 1982.PubMedGoogle Scholar
  38. 38.
    Toth, T. E. Trypsin-enhanced replication of neonatal calf diarrhea coronavirus in bovine embryonic lung cells. Am. J. Vet. Res. 43:967–972; 1982.PubMedGoogle Scholar
  39. 39.
    Tyrrell, D. A. J.; Almeida, J. D.; Berry, D. M.; Coronaviruses. Nature 220:650; 1968.Google Scholar
  40. 40.
    Tryrrell, D. A. J.; Almeida, J. D.; Cunningham, C. H., et al. Coronaviridae. Intervirology 5:76–82; 1975.PubMedGoogle Scholar
  41. 41.
    Tryrrell, D. A. J.; Alexander, J. D.; Almeider, J. D., et al. Coronaviridae: second report. Intervirology 10:321–328; 1978.PubMedGoogle Scholar
  42. 42.
    Vautherot, J. Plaque assay for titration of bovine enteric coronavirus. J. Gen. Virol. 56:451–455; 1981.PubMedGoogle Scholar
  43. 43.
    Weller, T. H.; Enders, J. F.; Robbins, F. C., et al. Studies on the cultivation of poliomyelitis viruses in tissue culture. I. The propagation of poliomyelitis viruses in suspended cell culture of various human tissues. J. Immunol. 69:645–671; 1952.PubMedGoogle Scholar

Copyright information

© Tissue Culture Association, Inc. 1988

Authors and Affiliations

  • Linda J. Saif
    • 1
  • Robert A. Heckert
    • 1
  • Kathy L. Miller
    • 1
  • Mohamed M. Tarek
    • 1
    • 2
  1. 1.Food Animal Health Research Program, Ohio Agricultural Research and Development CenterOhio State UniversityWooster, Ohio
  2. 2.Department of Parasitology and Microbiology, Faculty of MedicineZagazig UniversityZagazigEgypt

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