Trypanosoma cruzi is the etiological agent of Chagas disease that infects more than seven million people in Latin America. The parasite is transmitted by triatomine insects, of which some species are often associated with palms. The establishment of oil palm plantations (Elaeis guineensis) in the Orinoco region (Colombia) has been rapidly growing, possibly constituting a new environment for the establishment and increase in triatomine populations. In this study, the potential of Rhodnius prolixus to colonize E. guineensis plantations and maintain T. cruzi transmission was assessed. Fieldwork was conducted in two areas located in the department of Casanare for sampling E. guineensis and Attalea butyracea palms, sampling for triatomines to determine their abundance and prevalence of T. cruzi infection. To assess T. cruzi transmission potential in the area, sylvatic and domestic mammals were sampled. Results showed that palm infestation with triatomines was higher in A. butyracea than in E. guineensis palms and T. cruzi infection in triatomines varied between habitats for one study area, but was constant in the other site. Trypanosoma cruzi-infected mammals in the E. guineensis plantations were mainly generalist rodents, suggesting that these mammals could have an important role in T. cruzi transmission in plantations. In conclusion, E. guineensis plantations in the Orinoco region are suitable habitats for R. prolixus and T. cruzi transmission.