Bat flies (Nycteribiidae and Streblidae) have been used to study co-evolutionary patterns between ectoparasites and bats. In the world, Nycteribiidae and Streblidae are represented by approximately 276 and 237 species, respectively. In regions such as the Orinoquia located in the north of South America (Colombia and Venezuela), the richness of bats is high (more than 100 documented species), but studies on Nycteribiidae and Streblidae are scarce and discontinuous. To contribute to the knowledge of ectoparasitic flies in the Orinoquia, records of flies and their interactions with bats were reviewed, including new records and associations using interaction networks. We documented 124 species of Streblidae and only 12 of Nycteribiidae for the Orinoquia in approximately 102 bat species reported in Colombia and Venezuela. New records for six species of bat flies in Colombia were found (Mastopteraguimaraesi, Noctiliostreblamaai, Paradyschiriaparvuloides, Trichobiusjubatus, Trichobiusparasiticus, and Basiliaferrisi) associated with six species of bats (Cynomopsplanirostris, Desmodusrotundus, Myotishandleyi, Molossusrufus, Noctilioalbiventris, and Phyllostomushastatus). The bat-ectoparasite interaction networks in the Orinoquia revealed a pattern of antagonistic relationships, with high specialization, modularity, and low connectivity and nesting. The identified networks are between bat fly species belonging to different ecomorphological groups with unique host species. This supports the idea of ecological niche partitioning among ectoparasitic bat flies and hosts. Our study expanded the knowledge of the distribution of some fly species and the associations with bat hosts in Colombia, by presenting morphological descriptions and new observations, which are key to understanding the ecology, diversity, and distribution of these species.