Bats are members of the mammalian fauna, known for their high species richness and diversity, and play a crucial role as reservoirs of emerging and reemerging zoonotic pathogens. Thirty-five bat species have been reported in the coastal ecoregion of Restinga, in the state of Rio de Janeiro. Trachops cirrhosus is a carnivore/insectivore, feeding primarily on insects, lizards, and frogs. Desmodus rotundus is a hematophagous bat, feeding on the blood of other mammals (domestic and wild). Nematodes of the genus Tricholeiperia have been reported in bats of the families Noctilionidae and Phyllostomidae. The objective of this study is to contribute to the detailed ultrastructure of T. leiperi by performing a detailed characterization using scanning electron and confocal microscopy, in addition to reporting parasitism in D. rotundus for the first time. A total of 11 T. cirrhosus and 33 D. rotundus specimens were captured over five nights using mist nets. After euthanasia, the recovered helminths were preserved in 70% ethanol and subsequently examined for their morphological characteristics. Eight T. cirrhosus specimens were parasitized by T. leiperi, while only two D. rotundus specimens were parasitized. The helminths were identified through morphological and morphometric taxonomic characteristics, such as the shape of the cephalic vesicle, the presence of cervical papillae, the division at the distal end of the spicules, the size and arrangement of the bursa rays, the conspicuous vulva flap, and the presence of tubercles on the tip of the female's tail. The observed morphological characteristics involve the species T. leiperi previously described in T. cirrhosus, and this is the first report of T. leiperi in D. rotundus. This finding highlights the importance of studying parasite biodiversity in hosts across different biomes, particularly in heavily impacted ecosystems such as Restinga.