Abstract
Chicken tracheal organ cultures (TOCs) provide a simple ex vivo system that makes use of transverse section of tracheal rings extracted from embryos or adult birds to perform classical virological techniques for virus isolation, propagation and titrations, alongside with gene-expression analysis and virus-host interaction studies. Most IBV strains replicate well in TOCs, thus conveniently allowing growth kinetics analysis. Viral replication is revealed by observation of ciliostasis as marker of infection in tracheas extracted from birds ex vivo, as well as in vitro analysis providing a reliable infection model and a useful tool for titration.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McGee ZA, Woods ML (1987) Use of organ cultures in microbiological research. Ann Rev Microbiol 41:291–300
Cherry JD, Taylor-Robinson D (1970) Large-quantity production of chicken embryo tracheal organ cultures and use in virus and mycoplasma studies. Appl Microbiol 19:658–662
Barjesteh N, Alkie TN, Hodgins DC et al (2016) Local innate responses to TLR ligands in the chicken trachea. Viruses 8:207
Cook JKA, Darbyshire JH, Peters RW (1976) The use of chicken tracheal organ cultures for the isolation and assay of infectious bronchitis virus. Arch Virol 50:109–118
Cummiskey JF, Hallum JV, Skinner MS et al (1973) Persistent Newcastle disease virus infection in embryonic chicken tracheal organ cultures. Infect Immun 8:657–664
Higgins PG, Ellis EM (1972) The isolation of influenza viruses. J Clin Pathol 25:521–524
Hodgson T, Casais R, Dove B et al (2004) Recombinant infectious bronchitis coronavirus Beaudette with the spike protein gene of the pathogenic M41 strain remains attenuated but induces protective immunity. J Virol 78:13804–13811
de Wit JJ, Cook JK (2014) Factors influencing the outcome of infectious bronchitis vaccination and challenge experiments. Avian Pathol 43:485–497
Malo A, de Wit S, Swart WAJM et al (2017) Alternative methods to compare safety of live-attenuated respiratory Newcastle disease vaccines in young chicks. Vet Rec 181:236
Ellis S, Keep S, Britton P et al (2018) Recombinant infectious bronchitis viruses expressing chimaeric spike glycoproteins induce partial protective immunity against homologous challenge despite limited replication in vivo. J Virol 92:e01473–e01418
Yacida S, Aoyam S, Takahashi N et al (1978) Plastic multiwell plates to assay avian infectious bronchitis virus in organ cultures of chicken embryo trachea. J Clin Microbiol 8:380–387
Jackwood MW, Jordan BJ, Roh HJ et al (2015) Evaluating protection against infectious bronchitis virus by clinical signs, ciliostasis, challenge virus detection, and histopathology. Avian Dis 59:368–374
Hennion RM (2015) The preparation of chicken tracheal organ cultures for virus isolation, propagation, and titration. Methods Mol Biol 1282:51–56. https://doi.org/10.1007/978-1-4939-2438-7_5
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Dowgier, G., Bickerton, E. (2020). The Preparation of Chicken Tracheal Organ Cultures and Their Application for Ciliostasis Test, Growth Kinetics Studies, and Virus Propagation. In: Maier, H., Bickerton, E. (eds) Coronaviruses. Methods in Molecular Biology, vol 2203. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0900-2_8
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0900-2_8
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0899-9
Online ISBN: 978-1-0716-0900-2
eBook Packages: Springer Protocols