Mixophyes are large ground-dwelling myobatrachid frogs from eastern Australia and New Guinea. We use analyses of allozyme frequencies, nucleotide sequences of mitochondrial DNA and morphology to define species boundaries in Mixophyes from the Wet Tropics World Heritage Area of northern Queensland. The molecular analyses identify a minimum of three species in the region. Morphometric and meristic analyses corroborate these distinctions. The existence of two of these species was not previously suspected, and they are formally described herein.