Toxoplasmosis is a zoonotic disease of global distribution that affects all warm-blooded animals. The purpose of this investigation was to determine the prevalence of T. gondii infection and identify the risk factors associated with its occurrence in domestic ruminants raised on the island of Fernando de Noronha, Brazil, and to confirm that cattle and sheep raised in Fernando de Noronha Island present statistically different T. gondii prevalence rates. Serum samples were collected from sheep (n=240) and cattle (n=140) for the detection of antibodies by indirect immunofluorescence. Samples were collected from all the animals on all the farms. Risk factors were analyzed by univariate analysis and logistic regression. The prevalence rate of positive sheep was 85.0% while that of cattle was 10.7%. A multivariate analysis revealed that the site of contact of sheep with felines was a risk factor. For cattle, the risk factors identified in this study were: extensive farming system, water source, more than three cats per farm, and the presence of rats in feed storage locations. The findings revealed a significant difference in the prevalence rates in sheep and cattle raised in this insular environment.