Antillean manatees (Trichechus manatus manatus) are aquatic mammals that inhabit marine waters from Central America to the northeastern region of Brazil, and they are an endangered species. Infection with Toxoplasma gondii through intake of water or food contaminated with oocysts has been reported among marine mammals. The present study aimed to evaluate the prevalence of antibodies to T. gondii in West Indian manatees living in captivity in northeastern Brazil. Serum samples from 55 West Indian manatees from three different captive groups were tested for T. gondii antibodies by means of the modified agglutination test using a cutoff of 1:25. The samples were screened at dilutions of 1:25, 1:50, and 1:500, and positive samples were end-titrated using twofold serial dilutions; antibodies were found in six Antillean manatees (10.9%) with titers of 1:50 in three, 1:500 in one, 1:3,200 in one, and 1:51,200 in one manatee. This study is the first report of T. gondii antibodies in captive Antillean manatees in Brazil.