Stomoxys calcitrans is a hematophagous dipteran. Several agents are used in biological control, including entomopathogenic nematodes (EPNs). Bioassay I involved an evaluation of the effect of Heterorhabditis bacteriophora HP88 and Heterorhabditis baujardi LPP7 on S. calcitrans larvae in different periods of exposure. Groups of 10 larvae were placed in Petri dishes and 200 EPNs/larva were added, which were divided into groups according to the exposure times of 2, 4, 6, 12, 24 and 48 hours. The purpose of Bioassay II was to evaluate the efficacy of the EPNs in infecting S. calcitrans larvae when they were isolated from stable fly larvae in Bioassay I. Groups of 10 larvae were placed in Petri dishes and 200 EPNs/larva were added. In bioassay I, H. bacteriophora caused mortality rates of 51.7, 83.3 and 91.7% in 12, 24 and 48 hours, respectively, while H. baujardi caused mortality rates of 9.3 (12h), 35 (24h) and 35% (48h). In Bioassay II, H. bacteriophora and H. baujardi resulted in mortality rates of 35% and 25%, respectively. It was concluded that the longest exposure times presented the highest larval mortality and that EPNs isolated from S. calcitrans are not efficient in controlling the larvae fly.