We genetically characterize rabies virus (RABV) strains isolated from human cases, domestic and wild animals during a human outbreak of bat-transmitted rabies in Augusto Correa municipality, Pará state, Brazilian Amazon in 2005. Partial nucleotide sequences of the N gene (491 bp) were obtained for all strains, and phylogenetic analysis grouped these into two major clades (Pará and Central-Southeast) and identified them as bat-related viruses genotype I, Desmodus rotundus antigenic variant 3 (AgV3). A molecular clock was used to estimate the time of emergence for each RABV isolate. The molecular data from this study suggest the association of vampire bats with human and domestic animal cases reported in the outbreak, the circulation of at least two predominant lineages in the Pará state, and also a geographic association to lineages dispersion.