Despite the potential importance of temporal separation for the coexistence of competing species, no study has found significant segregation at the circadian level between jaguar (Panthera onca) and puma (Puma concolor) in sympatry. Using data from camera trap surveys (wet and dry seasons), we have evaluated the activity patterns of both species and their potential prey at four areas in the dry forest of the Bolivian Chaco. We tested if temporal separation existed between these two species, and if their activity was related to that of a particular prey. At most sites, activity patterns of jaguar and puma did not vary significantly between seasons, except for puma at one site. There were no differences between sexes for any cat species at any site. At three sites we found statistically significant differences in the activity patterns of jaguar and puma, as they showed a clear temporal segregation. None of them followed the activity patterns of any particular prey species across sites. The latter suggests that segregation is influenced by avoidance behaviour between the two felid species. Therefore, temporal separation may be an important behavioural factor promoting the coexistence of jaguar and puma in some areas of this dry forest.