Advertisement calls can be used to aid solving taxonomic problems and understanding the evolution of certain groups. In this study, the advertisement call of Eleutherodactylus bilineatus is described. It is composed by two different notes with a total duration of 0.529-4.241 seconds and dominant frequency of 1.72-3.45 kHz. Additionally, new data is provided on the geographical distribution of Eleutherodactylus bilineatus and the most inland record for this species.
Keywords: Atlantic Forest; Holoadeninae; bioacoustics; range extension; vocalization.