Nycteribiidae encompasses a specialized group of wingless blood-sucking flies that parasitize bats worldwide. Such relationships are frequently species- or genus-specific, indicating unique eco-evolutionary processes. However, despite this significance, comprehensive studies on the relationships of these flies with their hosts, particularly in the New World, have been scarce. Here, we provide a detailed description of the parasitological patterns of nycteribiid flies infesting a population of Myotis lavali bats in the Atlantic Forest of northeastern Brazil, considering the potential influence of biotic and abiotic factors on the establishment of nycteribiids on bat hosts. From July 2014 to June 2015, we captured 165 M. lavali bats and collected 390 Basilia travassosi flies. Notably, B. travassosi displayed a high prevalence and was the exclusive fly species parasitizing M. lavali in the surveyed area. Moreover, there was a significant predominance of female flies, indicating a female-biased pattern. The distribution pattern of the flies was aggregated; most hosts exhibited minimal or no parasitism, while a minority displayed heavy infestation. Sexually active male bats exhibited greater susceptibility to parasitism compared to their inactive counterparts, possibly due to behavioral changes during the peak reproductive period. We observed a greater prevalence and abundance of flies during the rainy season, coinciding with the peak reproductive phase of the host species. No obvious correlation was observed between the parasite load and bat body mass. Our findings shed light on the intricate dynamics of nycteribiid-bat interactions and emphasize the importance of considering various factors when exploring bat-parasite associations.