SUMMARY Leptospirosis is the major infectious disease on Reunion Island but little is known about the animal reservoir. We conducted a wide-ranging survey that included samples from 574 animals belonging to 12 species. The seroprevalence and prevalence of renal carriage varied greatly depending on the species, with the highest seroprevalence (79·5%) found in Norway rats, and the lowest (13·2%) in tenrecs. The renal carriage rate ranged from 84·6% in mice to 0% in tenrecs. Our results suggest that rodents are the most important reservoirs of leptospirosis on Reunion Island. The epidemiological role that animals play in human infection is discussed. For the first time, we quantified the renal concentration of leptospires in ten naturally infected mammals. The history of Reunion Island colonization probably explains why the circulating Leptospira serogroups were similar to those found in Europe. Our study provides evidence that will help implement preventive measures against this zoonosis.