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<i>Trypanosoma rangeli</i> is a generalist hemoflagellate that infects mammals and is transmitted by triatomines around Latin America. Due to its high genetic diversity, it can be classified into two to five lineages. In Brazil, its distribution outside the Amazon region is virtually unknown, and knowledge on the ecology of its lineages and on host species diversity requires further investigation. Here, we analyzed 57 <i>T. rangeli</i> samples obtained from hemocultures and blood clots of 1392 mammals captured in different Brazilian biomes. The samples were subjected to small subunit (SSU) rDNA amplification and sequencing to confirm <i>T. rangeli</i> infection. Phylogenetic inferences and haplotype networks were reconstructed to classify <i>T. rangeli</i> lineages and to infer the genetic diversity of the samples. The results obtained in our study highlighted both the mammalian host range and distribution of <i>T. rangeli</i> in Brazil: infection was observed in five new species (<i>Procyon cancrivorous</i>, <i>Priodontes maximum</i>, <i>Alouatta belzebul</i>, <i>Sapajus libidinosus</i>, and <i>Trinomys dimidiatus</i>), and transmission was observed in the Caatinga biome. The coati (<i>Nasua nasua</i>) and capuchin monkey (<i>S. libidinosus</i>) are the key hosts of <i>T. rangeli</i>. We identified all four <i>T. rangeli</i> lineages previously reported in Brazil (A, B, D, and E) and possibly two new genotypes.