Original Research

Propagation of avian influenza virus in embryonated ostrich eggs

Agnes T. Laleye, Modupeore Adeyemi, Celia Abolnik
Onderstepoort Journal of Veterinary Research | Vol 89, No 1 | a2011 | DOI: https://doi.org/10.4102/ojvr.v89i1.2011 | © 2022 Agnes T. Laleye, Modupeore Adeyemi, Celia Abolnik | This work is licensed under CC Attribution 4.0
Submitted: 07 March 2022 | Published: 08 December 2022

About the author(s)

Agnes T. Laleye, Department of Production Animal Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa; and, National Veterinary Research Institute, Vom, Nigeria
Modupeore Adeyemi, Department of Production Animal Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa
Celia Abolnik, Department of Production Animal Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa


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Abstract

Influenza A viruses (IAVs) are typically isolated and cultured by successive passages using 9- to 11-day-old embryonated chicken eggs (ECEs) and in 14-day old ECEs for virus mutational studies. Real-time reverse transcription-polymerase chain reaction tests (RT-PCRs) are commonly used for IAV diagnosis, but virus isolation remains invaluable in terms of its high sensitivity, providing viable isolates for further studies and the ability to distinguish between viable and nonviable virus. Efforts at isolating ostrich-origin IAVs from RT-PCR positive specimens using ECEs have often been unsuccessful, raising the possibility of a species bottleneck, whereby ostrich-adapted IAVs may not readily infect and replicate in ECEs, yet the capacity of an ostrich embryo to support the replication of influenza viruses has not been previously demonstrated. This study describes an optimised method for H5 and H7 subtype IAV isolation and propagation in 28-day old embryonated ostrich eggs (EOEs), the biological equivalent of 14-day old ECEs. The viability of EOEs transported from breeding sites could be maximised by pre-incubating the eggs for 12 to 14 days prior to long-distance transportation. This method applied to studies for ostrich-adapted virus isolation and in ovo studies will enable better understanding of the virus-host interaction in ostriches and the emergence of potentially zoonotic diseases.

Keywords

avian influenza virus; propagation; ostrich; embryonated eggs; protocol

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