Hunting dogs have probably a higher level of exposure to Neospora caninum Dubey, Carpenter, Speer, Topper et Uggla, 1988 and Toxoplasma gondii Nicolle et Manceaux, 1908 than other canine populations for their different lifestyle. The aim of our survey was to determine the seroprevalence of N. caninum and T. gondii in hunting dogs from southern Italy and assess risk factors related to these protozoan infections. Blood samples were collected from 398 hunting dogs (19 different breeds, aged from 5 month to 14 years). The sera were screened by indirect fluorescence antibody test; a titre greater or equal 50 was considered positive. Antibodies to N. caninum and T. gondii were detected in 59 (15%) dogs with titres from 50 to 3 200 and in 94 (24%) dogs with titres from 50 to 1 600, respectively, with co-infection in 25 (6%) dogs. Statistical difference (p less or equal 0.05) was found only for infection with T. gondii between two age groups: 2–4 years (16%) and 4–7 years (33%); other observed characteristics were without statistical significance. Our results suggest that the hunting dogs could play an important role in the transmission cycle of N. caninum between wild animals and livestock. This is the first detection of antibodies to T. gondii in hunting dogs in Italy.