American tegumentary leishmaniasis is endemic to the Espírito Santo State, Brazil, where it is widely distributed. The composition of the phlebotomine sand fly fauna in an American tegumentary leishmaniasis focus was determined by monthly sampling, using Shannon light traps in an Atlantic Forest reserve and adjacent habitat that had been modified by human activity. Seasonal fluctuations in numbers of the most abundant species were also monitored from June 2004 to May 2006. Of the 6,176 specimens collected, 47.4% were captured in the forest and 52.6% in the disturbed habitat. Although Lutzomyia davisi (60.8%) predominated in specimens from the forest, those captured near human dwellings consisted almost entirely of Lu. choti (72%) and Lu. intermedia (24.3%). All three species occurred throughout the year. Based on our findings, Lu. intermedia probably acts as the principal domestic Leishmania vector in the study area.