The sand fly fauna in Itatiaia National Park, Rio de Janeiro, Brazil, was investigated in different habitats ranging from sylvatic areas to those altered by human activity related to ecotourism, specifically identifying species that have been suggested as potential leishmaniasis vectors. Sand flies were captured from June 2002 to March 2004, using CDC light traps and Shannon traps. A total of 1,256 sand fly specimens were captured, from species belonging to genera Lutzomyia and Brumptomyia: Brumptomyia guimaraesi, B. troglodytes, Lutzomyia (Lutzomyia) amarali, L. lanei, L. migonei, L. sallesi, L. edwardsi, L. tupynambai, L. (Pintomyia) pessoai, L. (P.) bianchigalatie, L. rupicola, L. (Psathyromyia) shannoni, L. pascalei, L. (Psychodopygus) matosi, L. (P.) davisi, L. (P.) hirsuta hirsuta, L. (P.) ayrozai, L. peresi, L. monticola, and L. misionensis. Worthy of special attention were four species that are considered potential vectors for cutaneous leishmaniasis in Brazil: L. ayrozai, L. hirsuta hirsuta, L. migonei, and L. davisi, representing 19.19% of the specimens captured in this study.