The Brazilian savannah-like area, the Cerrado region, covers large areas of the country and provides a habitat for a multitude of different animal species. The giant anteater (Myrmecophaga tridactyla) is geographically widespread and one of the typical inhabitants of the Cerrado. They are listed as vulnerable by the International Union for Conservation of Nature and Natural Resources. A population loss of at least 30 % over the past 10 years has been estimated based on local extinctions, habitat loss and deaths caused by fires, roadkills and hunting. Little is known about ecological and in particular parasitic conditions of this highly specialised insectivore species. During September and November 2010 we examined three roadkilled giant anteater for the presence of metazoan ecto- and endoparasites. Besides the cestode species Oochoristica tetragonocephala and the tick species Amblyomma nodosum, we found for the first time the flea Tunga penetrans. Beside morphological flea species identification, we compared a fragment of the 16S rRNA gene of eggs, a molecular method utilised for the first time in this combination. The identification of T. penetrans in M. tridactyla represents a new host record and expands the host and distribution range of the zoonotic flea species.