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Lyssavirus is a genus that causes infectious disease transmit by bats transmit, which results in economic losses in livestock and public health problems. From 2005 to 2019, more than 49 thousand cases of the disease were registered in animals in Brazil, with 3418 registered in Mato Grosso (MT). The lack of information on the genetic diversity and distribution of the rabies virus in MT was the motivation for carrying out this study. A total of 117 samples of brain tissue from cattle, horses, donkeys, mules and sheep from 29 municipalities in the state of MT and one municipality in Rondônia were used. Direct immunofluorescence and/or biological tests performed from 2014 to 2021 indicated that all samples were positive for the disease. RNA was extracted and molecular analysis was performed using RT-PCR for the N gene. Of the 117 samples analyzed, 50 were amplified by RT-PCR, purified and sequenced. The samples showed 93.13%-100% identity with the rabies virus. The sequences were submitted to phylogenetic analysis that resulted in a tree of four clades; these were genetically grouped into distinct regions within the Desmodus rotundus lineage. The results of the geolocation of clades will be useful to guide monitoring, control and health surveillance programs in MT., (Copyright © 2022. Published by Elsevier B.V.)