Twenty-seven rickettsiae were isolated and/or detected from 100 Amblyomma variegatum ticks collected on Guadeloupe in the French West Indies. In this study, the polymerase chain reaction procedure appeared to be more sensitive in detecting rickettsiae in ticks than the shell-vial technique. Sequencing a portion of the outer membrane protein A-encoding gene showed that these rickettsiae appeared to be identical to Rickettsia africae, a member of the spotted fever group rickettsiae recently described as an agent of African tick-bite fever occurring in sub-Sahelian Africa. A high seroprevalence of antibodies to R. africae was demonstrated among mammals, particularly humans, cattle, and goats. These results and a recently reported case of an infection due to R. africae on Guadeloupe demonstrate that R. africae is present on this island. Although this disease has been underdiagnosed there, it may be frequent and may exist on other Caribbean islands where A. variegatum has propagated dramatically over recent years.