Rabies is a disease caused by a virus belonging to the family Rhabdoviridae, genus Lyssavirus (RABV), and is characterized as a progressive and acute encephalitis that presents high lethality. Between 2002 and 2012, 2149 wild animals were found positive for RABV in Brazil where 79% were associated with the aerial cycle of the disease. As part of the active surveillance of rabies in Brazil, the objective of this study was to investigate the presence of RABV infection in bats captured in the metropolitan region of Rio de Janeiro city. Forty-four bats were tested for RABV at Institute Jorge Vaitsman (IJV), reference laboratory for rabies in Rio de Janeiro State. Of the individuals captured and sent for the diagnosis of rabies, a total of 15 were identified as being hematophagous, all of them belonging to the species Desmodus rotundus. Twenty-four individuals were considered as frugivorous, four as insectivores and one individual (Lonchophylla peracchii) identified as having a pollen/ nectar feeding habit. All specimen sent to RABV diagnosis presented negative results. Considering the active surveillance for RABV in Brazil, studies of fauna survey with serological characterization are important in the strengthening of epidemiological surveillance and protection of human life.