The aim of the present study was to determine the prevalence of Babesia bovis and Babesia bigemina in dairy cattle from São Luis Island in the state of Maranhão, Brazil. A total of 281 blood samples were collected. In total, 275 (97.9%) animals were B. bovis-reactive and B. bigemina-reactive in the Enzyme-Linked Immunosorbent Assay (ELISA). The microscopy examination detected 22 (7.8%) animals that were positive for Babesia sp. and the Polymerase Chain Reaction (PCR) analysis showed that 91 animals (32.38%) and 23 animals (8.18%) were positive for B. bovis and B. bigemina, respectively, while 17 animals (6.04%) were co-infected. There is a high level of transmission of these protozoa in Maranhão, and the animals were naturally exposed. Therefore, it is possible to characterize the island as enzootic stability for babesiosis, indicat-ing a risk of financial losses when susceptible animals are introduced from areas of enzootic instability or free regions of B. bovis and B. bigemina.Título em português: Avaliação sorológica, parasitológica e molecular de Babesia bovis e Babesia bigemina no gado do Estado do Maranhão