BgV replication in vertebrate animal models.

Groups of three eggs were inoculated with three doses of BgV intravenously (105.7, 104.7 and 103.7 TCID50 IU/egg, respectively), incubated at 33–35°C for five days and homogenised. The homogenates were titrated on C6/36 cells and the resulting virus titres were analysed by fixed-cell ELISA. The eggs injected at the higher doses had traces of virus present but at levels suggesting they resulted from leftover inoculum rather than virus replication. The other animal models were inoculated with 106 TCID50 IU of BgV and sampled for blood on days 1–5 and 14 post-inoculation (sparrows) or on days 1, 3, 5, 7, and 28 post-inoculation (frogs, toads, snakes). The blood samples were diluted 1:5 and 1:50 and inoculated on C6/36 cells which were incubated for two days and analysed for virus by fixed-cell ELISA.