Introduction. Canine visceral leishmaniasis in endemic areas of Colombia could be a public health risk factor given the zoonotic nature of the disease. Ninety-six human cases of visceral leishmaniasis were reported in Colombia in 2004, 5 of them in Huila, where Lutzomyia longipalpis has been incriminated as the main vector species.Objective. To determine the prevalence of IgG antibodies against Leishmania chagasi in dogs from the sector 8 of the city of Neiva and the from the towns of Villavieja, Algeciras, Palermo and Rivera located in Huila, Colombia.Materials and methods. An epidemiological survey was carried out in 610 dogs, which included clinical examination and venopuncture for obtaining blood samples. Authorization was obtained from the dog owners. The sera were analyzed by the ELISA test with promastigotes of the L. chagasi strain MHOM/CO84/CL044B as antigen.Results. The canine population had an average age of 2.5 years; 67.3% of the dogs were males and the cross–bred animals were the most prevalent constituting 85% of those samples.On clinical examination the main signs were onicogriphosis 24.3%, lymphadenitis 10% and skin lesions 5%. The presence of antibodies was observed in 28.1% of the dogs from sector 8 of Neiva, 28% in Villavieja, 14.9% in Rivera, 10% in Palermo and 5.1% in Algeciras. An average ratio of five people cohabitating per seropositive dog was observed.Conclusions. The results reflect the exposure to Leishmania chagasi infection of dogs living in both urban and rural environments in the studied zones, and should encourage health authorities to carry out control measures to prevent the spread of this zoonotic disease.