In nocturnal chorusing anurans, mate choice has long been considered to be based exclusively on the highly conspicuous male calls. However an additional visual cue, the pulsing male vocal sac, has recently been shown to increase male attractiveness. In several species, including the European treefrog Hyla arborea, this organ is characterized by a bright orange colouration. In the present study, analysis of the pigments extracted from the vocal sacs of European treefrogs revealed the colouration of these organs to be based on several pigments, including three carotenoids, which are highly costly immune function enhancing molecules. This result raises the important question as to whether the carotenoid-based colouration of the vocal sac in H. arborea could constitute an additional visual cue used by females to discriminate male quality.