Pará State comprises 26% of Brazilian Amazon Region where a large diversity of arboviruses has been described. This study aimed to assess the prevalence and distribution of hemagglutination-inhibition (HI) that detect antibodies against four units to Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), Mayaro virus (MAYV) and Mucambo virus (MUCV), a subtype of the Venezuelan equine encephalitis virus (VEEV), in 2,191 serum samples of horses, cattle, sheep and water buffaloes in Pará State, Brazil. The goal was to identify the prevalence of antibodies in these domestic farm animals to determine which arboviruses are circulating and determine which farm animal is the most sensitive for detecting Alphavirus. Antibodies against all investigated arboviruses were detected in almost all animals species studied. Our results indicated that domestic herbivores are susceptible to the tested arboviruses and evidence of active Alphavirus in farm animals in the Brazilian Amazon. An analysis of HI antibody prevalence by animal species indicated significant difference between horses and water buffaloes, cattle and sheep. The horses showed higher prevalence of antibodies in heterotypic reactions showing to be the best species of domestic farm animal to serve as sentinel to detect the movement of arboviruses in the Brazilian Amazon.