Virus isolation
The supernatants of PiAdV-A and PiCV positive samples from a pigeon flock were inoculated into SPF embryonated chicken eggs. All samples were passaged 5 times in SPF chicken eggs. It was not possible to propogate PiAdV-A virus from the PiAdV-A PCR positive sample in eggs, but PiCV virus was propagated from the PiCV PCR positive sample in the SPF embryonated chicken eggs and caused extensive damage to chicken embryos regardless of the inoculation route (chorioallantoic cavity and yolk sac) (Figure.3). PiAdV-A and PiCV positive samples did not lead to any propogation of virus in primary CEFCs, but both samples caused extensive rounding, clumping and detatchment of cells in the CEKC cultures 48-72 hours post-inoculation (Figure. 4).
FIGURE 3. a-b-c) Extensive malformation and hemorrhaging was caused in chicken embryos by PiCV (TR/SKPC20) and compared with normal chicken embryo (d). Livers with multiple petechial haemorrhages (black arrows)