The goal of this study was to investigate anti-Rickettsia spp. antibodies in sera of domestic chickens (Gallus gallus domesticus) of extensive breeding in Cerro Largo county, considered an endemic area for spotted fever in the State of Rio Grande do Sul, Brazil. Three hundred blood samples were collected and anti-Rickettsia spp. antibodies were evaluated by indirect immunofluorescence assay (IFA) in the sera obtained. The occurrence of anti-Rickettsia spp. antibodies detected in this study was 1.33% (4/300), with endpoint titers ranging from 64 to 256 for Rickettsia rickettsiii, R. parkeri and/or R. bellii. The results suggest these domestic chickens do not participate as a reservoir and/or amplifying host in the epidemiology of spotted fever in that endemic area. The present study consists in the first serological survey in Gallus gallus domesticus to Rickettsiae-spotted fever group in Brazil.