Toxoplasma gondii is the etiological agent of toxoplasmosis, a widespread zoonosis that affects several homeothermic animals, including humans. This disease causes serious health problems, such that 10% of infected individuals develop clinical manifestations. Some studies on indigenous human populations have indicated variations in seroprevalence from 10.6% to 80.4% in such populations in different regions of Brazil and in other countries like Venezuela and Malaysia. To date, there have been no studies regarding the prevalence of anti-T. gondii antibodies in Haliti-Paresí Indians living in Campo Novo do Parecis, Mato Grosso, Brazil. Our objective here was to determine the frequency of occurrence of antibodies against this protozoon in nine Haliti-Paresí villages by correlating seroprevalence with locations and variables. Serodiagnoses were made using the indirect immunofluorescence antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) in the Laboratory for Toxoplasmosis and Other Protozoan Diseases of IOC/Fiocruz. It was considered that samples tested positive for T. gondii infection if IgG/IgM antibodies against this protozoon were detected through serodiagnosis using either IFAT or ELISA. Among the 293 samples analyzed, 66.9% presented anti-T. gondii IgG and 3.4% presented anti-T. gondii IgM. It was observed that there were no statistically significant differences in frequency of antibody occurrence among infected individuals, based on sex, schooling or occupation/activities. However, there were statistical differences based on age and villages. The prevalence observed in this study is in agreement with values found in other studies on indigenous populations in Latin America. Like among other such populations, the Haliti-Paresí villages are located close to forests and the individuals have domestic cats as pets, are involved in hunting and farming and consume water directly from water accumulation sources. These factors might cause exposure to T. gondii tissue cysts and oocysts.