Four hundred forty-eight blood plasma samples from free-living birds of prey from Berlin and the Brandenburg area in eastern Germany were tested for antibodies against New-castle disease virus (NDV), falcon herpesvirus (FHV), owl herpesvirus (OHV), and Chlamydia psittaci. Antibodies to NDV were detected in 6 (2%) of 346 tested diurnal birds of prey, whereas none of the owls (n = 55) was positive. The positive samples originated from two common buzzards (Buteo buteo), three ospreys (Pandion haliactus) and one marsh harrier (Circus aeruginosus). Titers varied between 1:8 and 1:32. Of 253 birds of prey one osprey (<1%) tested positive for antibodies to FHV with low titer of 1:6. This is the first detection of antibodies against FHV in an osprey. Furthermore, antibodies against OHV could be found in one tawny owl (Strix aluco) and one common buzzard (2 of 253, 1%) with low titers of 1:6. Of 422 birds of prey 267 (63%) tested positive for antibodies to Chlamydia psittaci with titers varying between 1:5 and 1: 256 which reflects the ubiquitous occurrence of Chlamydia psittaci in these birds of prey.