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. 1988;11(3):139–145. doi: 10.1007/BF01404267

Cell culture propagation of bovine coronavirus

Linda J Saif 1, Robert A Heckert 1, Kathy L Miller 1, Mohamed M Tarek 1,2
PMCID: PMC7088767  PMID: 32214596

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

References

  • 1.Akashi H., Inaba Y., Miura Y., et al. Properties of a coronavirus isolated from a cow with epizootic diarrhea. Vet. Microbiol. 1980;5:265–276. [Google Scholar]
  • 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. 1981;67:367–370. doi: 10.1007/BF01314841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 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. 1986;37:253–256. [PubMed] [Google Scholar]
  • 4.Baron S., McKerlie M. L. Broadly active inhibitor of viruses spontaneously produced by many cell types in culture. Infect. Immun. 1981;32:449–453. doi: 10.1128/iai.32.2.449-453.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Bosgiraud C., Nicolas J. A. Pneumonie a coronavirus chez les bovine: A propos de deux observations. Rec. Med. Vet. 1986;162:1085–1086. [Google Scholar]
  • 6.Bridger J. C., Caul E. O., Egglestone S. I. Replication of an enteric bovine coronavirus in intestinal organ cultures. Arch Virol. 1978;57:43–51. doi: 10.1007/BF01315636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 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.
  • 8.Dea S., Roy R. S., Begin M. E. Bovine coronavirus isolation and cultivation in continuous cell lines. Am. J. Vet. Res. 1980;41:30–38. [PubMed] [Google Scholar]
  • 9.Debiaggi M., Perduca M., Romero E., et al. Phosphatidyl-serine inhibition of OC43 and NCDCV coronavirus infectivity. Microbiologica. 1985;8:313–317. [PubMed] [Google Scholar]
  • 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.
  • 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. 1976;25:355–370. doi: 10.1016/0014-4800(76)90045-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 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. 1981;54:91–102. doi: 10.1099/0022-1317-54-1-91. [DOI] [PubMed] [Google Scholar]
  • 13.Hirano N., Sada Y., Tuchiya K., et al. Plaque assay of bovine coronavirus in BEK-1 cells. Jpn. J. Vet. Sci. 1985;47:679–681. doi: 10.1292/jvms1939.47.679. [DOI] [PubMed] [Google Scholar]
  • 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. 1981;32:454–457. doi: 10.1128/iai.32.2.454-457.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Inaba Y., Sato K., Kurogi H., et al. Replication of bovine coronavirus in cell line BEK-1 culture. Arch. Virol. 1976;50:339–342. doi: 10.1007/BF01317959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Kaye H. S., Yarbrough W. B., Reed C. J. Calf diarrhoea coronavirus. Lancet. 1975;II:509. doi: 10.1016/S0140-6736(75)90591-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 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. 1981;12:243–251. [PubMed] [Google Scholar]
  • 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. 1979;40:1476–1478. [PubMed] [Google Scholar]
  • 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.
  • 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. 1980;290:623–626. [PubMed] [Google Scholar]
  • 21.McNulty M. S., Bryson D. G., Allan G. M., et al. Coronavirus infection of the bovine respiratory tract. Vet. Microbiol. 1984;9:425–434. doi: 10.1016/0378-1135(84)90063-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 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. 1973;34:145–150. [PubMed] [Google Scholar]
  • 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.
  • 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. 1980;41:1039–1044. [PubMed] [Google Scholar]
  • 25.Reynolds D. J. Coronavirus replication in the intestinal and respiratory tracts during infection of calves. Ann. Rech. Vet. 1983;14:445–446. [PubMed] [Google Scholar]
  • 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. 1986;47:1426–1432. [PubMed] [Google Scholar]
  • 27.Sato K., Inaba Y., Kurogi H., et al. Physico-chemical properties of calf diarrhea coronavirus. Vet. Microbiol. 1977;2:73–81. [Google Scholar]
  • 28.Sharpee R. L., Mebus C. A., Bass E. P. Characterization of a calf diarrheal coronavirus. Am. J. Vet. Res. 1976;37:1031–1041. [PubMed] [Google Scholar]
  • 29.Siddell S., Wege H., Ter Melullen V. The biology of coronaviruses. J. Gen. Virol. 1983;64:761–776. doi: 10.1099/0022-1317-64-4-761. [DOI] [PubMed] [Google Scholar]
  • 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. 1972;33:1147–1156. [PubMed] [Google Scholar]
  • 31.Storz J., Rott R., Kaluza G. Enhancement of plaque formation and cell fusion of an enteropathogenic coronavirus by trypsin treatment. Infect. Immun. 1981;31:1214–1222. doi: 10.1128/iai.31.3.1214-1222.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 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. 1976;1:65–69. [Google Scholar]
  • 33.Sturman L. S., Homes K. V. The molecular biology of coronaviruses. Adv. Virus Res. 1983;28:35–112. doi: 10.1016/S0065-3527(08)60721-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Takahashi E., Inaba Y., Sato K., et al. Epizootic diarrhoea of adult cattle associated with a coronavirus-like agent. Vet. Microbiol. 1980;5:151–154. [Google Scholar]
  • 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. [PubMed]
  • 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.
  • 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. 1982;33:170–182. doi: 10.1016/S0034-5288(18)32331-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Toth T. E. Trypsin-enhanced replication of neonatal calf diarrhea coronavirus in bovine embryonic lung cells. Am. J. Vet. Res. 1982;43:967–972. [PubMed] [Google Scholar]
  • 39.Tyrrell D. A. J., Almeida J. D., Berry D. M. Coronaviruses. Nature. 1968;220:650. [Google Scholar]
  • 40.Tryrrell D. A. J., Almeida J. D., Cunningham C. H., et al. Coronaviridae. Intervirology. 1975;5:76–82. doi: 10.1159/000149883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Tryrrell D. A. J., Alexander J. D., Almeider J. D., et al. Coronaviridae: second report. Intervirology. 1978;10:321–328. doi: 10.1159/000148996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Vautherot J. Plaque assay for titration of bovine enteric coronavirus. J. Gen. Virol. 1981;56:451–455. doi: 10.1099/0022-1317-56-2-451. [DOI] [PubMed] [Google Scholar]
  • 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. 1952;69:645–671. [PubMed] [Google Scholar]

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