Antennae are among the most elaborate sensory organs in adult flies, and they provide rich information for phylogenic studies. The antennae of five out of eight species of Gasterophilus Leach (G. haemorrhoidalis (Linnaeus), G. intestinalis (De Geer), G. nasalis (Linnaeus), G. nigricornis (Loew) and G. pecorum (Fabricius)), were examined using scanning electron microscopy. The general morphology, including distribution, type, size, and ultrastructure of antennal sensilla were presented, and the definition of auriculate sensilla and sensory pits were updated and clarified. Eighteen antennal characters were selected to construct the first species-level phylogeny of this genus. The monophyly of Gasterophilus was supported by the presence of coeloconic sensilla III on the antennal arista. The species-level cladogram showed G. pecorum branching off at the base, and the remaining species forming the topology (G. intestinalis+ (G. haemorrhoidalis+ (G. nasalis+ G. nigricornis))). Our research shows the importance of the antennal ultrastructure as a reliable source for phylogenetic analysis.