Herpesvirus (HV) was detected using PCR in the organs of eight of 55 wild owls (14.5%) from seven species that were found dead in various locations in Slovenia between 1995 and 2015. HV was detected in three species: the Eurasian eagle owl (Bubo bubo), Ural owl (Strix uralensis), and long-eared owl (Asio otus). Phylogenetic analysis of partial DNA polymerase gene nucleotide sequences showed that the detected HVs are similar to the avian and mammal alphaherpesviruses. Two sequences were very similar to known bird HV sequences. One sequence was identical to the columbid herpesvirus 1 (CoHV1) sequence, and the other was very similar to the gallid herpesvirus 2 (GaHV2) sequence. The phylogenetic tree revealed a lower similarity of the other six analyzed Slovenian sequences with the sequences of alphaherpesviruses of birds and mammals. This is the first study to report the detection of different HVs in owls.