Harlequin frogs, genus Atelopus, communicate at high frequencies despite most species lacking a complete tympanic middle ear that facilitates high frequency hearing in most anurans and other tetrapods. Here we test whether Atelopus are better at sensing high frequency acoustic sound compared to other eared and earless species in the Bufonidae family, determine whether middle ear variation within Atelopus affects hearing sensitivity, and test potential hearing mechanisms in Atelopus. We determine that at high frequencies (2000–4000 Hz) Atelopus are 10–34 dB more sensitive than other earless bufonids but are relatively insensitive to mid-range frequencies (900–1500 Hz) compared to eared bufonids. Hearing among Atelopus species is fairly consistent, evidence that the partial middle ears present in a subset of Atelopus species do not convey a substantial hearing advantage. We further demonstrate that Atelopus hearing is not likely facilitated by vibration of the skin overlying the normal tympanic membrane region or the body lung wall, leaving the extratympanic hearing pathways in Atelopus enigmatic. Together these results show Atelopus have sensitive high frequency hearing without the aid of a tympanic middle ear and prompt further study of extratympanic hearing mechanisms in anurans.

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