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Leptospirosis is an emerging zoonotic disease caused by bacteria of the genus Leptospira. Wild animals may present acute disease or become chronic hosts. The present study aimed to identify Leptospira spp. infection and determine circulating serogroups in free-ranging sambar deer (Rusa unicolor) in a fragment of peri-urban tropical forest in northeastern Brazil. Blood samples were collected and subjected to microscopic agglutination testing (MAT) and PCR analysis (genes lipL32 and secY). Anti-Leptospira antibodies were detected in 60% of the animals tested, with serogroups Cynopteri (33.4%), Hebdomadis (22.2%) and Grippotyphosa (22.2%) identified as most prevalent. DNA corresponding to the pathogenic species Leptospira interrogans was detected in 2/15 (13.3%) of the samples tested. Sambar deer experience a high level of exposure to Leptospira spp. in our epidemiological setting. It is important to emphasize the implementation of effective measures (i.e., maintaining habitats and reducing human contact) for the conservation of endangered species., (© 2022. EcoHealth Alliance.)