Simple SummaryThe domestic donkey is an endangered equid species. There are 199 donkeys in the Special Nature Reserve “Zasavica”, out of the estimated 1000 that are assumed to exist in Serbia. The aim of this study was to assess the exposure of donkeys in the Special Nature Reserve “Zasavica” to some of the most significant viral pathogens and establish the seroprevalence of equine infectious anemia, African horse sickness, equine herpesvirus type 1 infection, equine influenza and equine viral arteritis. In tested blood samples, antibodies against equine herpesvirus type 1 and equine influenza virus (subtype H3N8) were found, while all samples tested negative for African horse sickness virus, equine infectious anemia virus and equine arteritis virus. To our knowledge, this study represents the first results of detection of these pathogens in the population of domestic donkeys in Serbia.AbstractThe paper presents the findings of specific antibodies in the blood sera of donkeys against the following viruses: equine infectious anemia virus (EIAV), African horse sickness virus (AHSV), equine herpesvirus type 1 (EHV-1), equine influenza virus subtype H3N8 (EIV) and equine arteritis virus (EAV). The analyses were conducted during the year 2022. From a total of 199 donkeys bred in “Zasavica”, blood was sampled from 53 animals (2 male donkeys and 51 female donkeys), aged 3 to 10 years. Specific antibodies against EIAV were not detected in any of the tested animals using the agar-gel immunodiffusion (AGID) assay. No specific antibodies against AHSV, tested by enzyme-linked immunosorbent assay (ELISA), or antibodies against EAV, tested by the virus neutralization test (VNT) and ELISA were detected in any of these animals. A positive serological result for EHV-1 was determined by the VNT in all animals, with antibody titer values ranging from 1:2 to 1:128, while a very low antibody titer value for EIV (subtype H3N8) of 1:16 was determined in 18 donkeys using the hemagglutination inhibition test (HI test).