Detectable levels of replication-competent virus in throat swabs, lung and nasal turbinate tissues post challenge were analysed. Quadruplicate, ten-fold serial dilutions were transferred to 96-well plates with Vero E6 cell culture monolayers and incubated for 1 h at 37 °C. Cell monolayers were washed prior to incubation for 5 days at 37 °C. Plates were then scored using the vitality marker WST8 and viral titres (Log10 TCID50/ml or /g) were calculated using the method of Spearman–Karber. Analyses were done at Viroclinics Xplore (Schaijk, The Netherlands).

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