Three auditory brainstem response (ABR) methods tested and compared in two anuran species.

Hearing sensitivity has been extensively investigated, often by measuring the auditory brainstem response (ABR). ABR measurements are relatively non-invasive, easy to reproduce, and allow the assessment of sensitivity when psychophysical data are difficult to obtain. However, the experimental methods differ greatly in respect to stimulation, which may result in different audiograms. We used three different methods in the same individual frogs: stimulating with brief tone bursts (tABR), long-duration tones (ltABR) and masked ABR (mABR), where transients are masked by a long-duration sinusoid, and the sensitivity is assessed by the difference between unmasked and masked ABR. We measured sensitivity in a range from 100 to 3500 Hz, and the resulting audiograms show two sensitivity peaks at 400-600 Hz and 1500-1600 Hz (both sensitive down to 30 dB re. 20 µPa). We found similar results below 1000 Hz, but when stimulating with long-duration tones, the sensitivity decreased more rapidly above this frequency. We showed that the frequency specificity of tone bursts becomes poorly defined with shorter duration at low frequencies. Comparisons between subjectively (visual inspection by researchers) and objectively (thresholds defined by signal-to-noise ratio) defined audiograms showed very little variation. In conclusion, the mABR method gave the most sensitive audiograms. The tABR method showed a similar audiogram when using relatively long-duration tone bursts (25 ms). The ltABR method is not a good choice for studying hearing thresholds above 1000 Hz because of the bias introduced by spike rate saturation in the nerve fibers and their inability to phase lock.