Trypanosomosis caused by Trypanosoma vivax has been a constraint for cattle production in the Bolivian lowlands, since it was introduced in 1996. Flooded areas like the Bolivian Pantanal have a suitable environment for the presence and transmission of Salivarian trypanosomes and farmers from that region often report trypanosomosis-like problems on their farms. The objective of the present study, therefore, was to characterize the epidemiology of bovine trypanosomosis in the Bolivian Pantanal. In order to achieve this objective, 202 cattle from the province of Angel Sandoval and 209 cattle from the province of German Busch were randomly sampled (the Pantanal is located in both provinces). Twenty-nine farms in both provinces were visited, the farmers interviewed, and biologic samples collected from their cattle. Samples were submitted for parasitological and PCR evaluation and the prevalence of bovine trypanosomosis was estimated for each province. Laboratory results were correlated with the sampled animals packed cell volume (PCV) and body condition (BC) scores and the observed T. vivax parasites measured for morphometry analysis. Results from this study show differences in morphometric measures between T. vivax parasites from each province. Differences between provinces were also observed in the T. vivax-related disease situation. While in Angel Sandoval the PCV and BC of T. vivax-affected animals were significantly lower than those of the T. vivax-negative animals, in German Busch no differences were observed in the PCV and BC of T. vivax-positive or negative animals. Animal prevalence of T. vivax in Angel Sandoval was 27.79% (95% CI: 14.52-44.28) and in German Busch was 19.03% (95% CI: 9.19-30.75). The T. evansi animal prevalence in each province was 0.99% (95% CI: 0.27-2.99) and 5.71% (95% CI: 2.43-12.19), respectively. Based on questionnaire and laboratory results, it was concluded that trypanosomosis is a primary constraint for cattle production in the Bolivian Pantanal.