Investigations have been carried out to elucidate the possible role of the donkey in the epidemiology of African horse sickness (AHS). These studies have shown that despite the absence of pyrexia or other observable clinical signs, donkeys become infected with virulent AHS virus serotype 4 (AHSV 4) and that they develop a viraemia which can persist for at least 12 days, albeit at a comparatively lower titre than that recorded for similarly infected ponies. AHSV 4 showed a similar tissue tropism in the pony and donkey but the virus appeared to replicate less efficiently in donkey tissues. The only gross pathological changes observed in the donkeys post mortem were increased fluid accumulation in the serosal lined compartments, particularly the peritoneal cavity, and petechial and ecchymotic haemorrhages on the left hepatic ligament. The absence of infectious virus or viral antigens in any of the tissues collected at 14 and 19 days post inoculation (dpi) from 6 experimental donkeys suggest that, though susceptible to infection, the donkey is unlikely to be a long term reservoir for AHSV. Although AHSV 4 was detected in all 6 donkeys following the primary inoculation, no virus could be isolated from blood collected from two donkeys subsequently challenged with a second virulent virus, AHSV 5. Data generated from virus neutralisation tests showed a second primary antibody response, against AHSV 5, in these donkeys at 12 dpi. In contrast, the boost in antibody levels detected from 5 dpi, as measured by ELISA, was probably due to an anamnestic response against the AHSV group-specific viral proteins. Homogenised spleen tissue, collected post mortem from a donkey 7 dpi with AHSV 4, caused a lethal, cardiac form of AHS when inoculated into a susceptible pony.