During a serological survey in 1996, a total of 601 dogs (group I) distributed all over the government controlled southern area of Cyprus was examined using an enzyme-linked immunosorbent assay (ELISA) for the presence of specific antibodies directed against soluble antigens of promastigote stages of Leishmania infantum. The overall seroprevalence rate in this group was 1.7%. A second group (group II) of dogs was selected from regions where seropositive dogs where determined within the first group. In group II specific anti-Leishmania antibodies could be detected in 30 of 301 dogs investigated (10%). The highest seroprevalence rates were found in the regions of Agios Georgios (26.2%) and Limnatis (12.2%). Ten parasite isolates from ten dogs (six with typical symptoms of canine leishmaniosis and four without symptoms) originating from five locations could be characterised by zymodeme analysis. All ten isolates were identified as L. infantum zymodeme MON 1.