Echinococcus oligarthrus is a tapeworm endemic to South America and widely distributed in the Amazon region. Its lifecycle is maintained by relationships between felids and their prey, mainly small sylvatic rodents, but humans can be infected occasionally. We report two female jaguarundis (Herpailurus yagouaroundi) harboring E. oligarthrus in southern Brazil. The felines were found road killed in periurban areas, and, during necropsy, the small intestine was examined. Visual inspection revealed helminths, which were submitted to microscopy and molecular examination. Morphologically, they were around 2.5 mm long, with four suckers and an armed scolex with two rows of hooks. Phylogenetic reconstruction using cytochrome c oxidase subunit I gene sequences placed samples from south Brazil in the same clade as all other E. oligarthrus samples, but as a sister group. Genetic distance gave similar results, resulting in a divergence of 0.087% between the samples described in this study and other samples. The geographic pattern of genetic diversity, as assessed by analysis of molecular variance, suggests that the divergency results from isolation by distance. This finding expands the geographic range of E. oligarthrus and brings new insights to help understand and prevent the zoonosis it causes.