Behavioral Responses to Ultrasonic Digital Music Inspired by Murine Courtship Songs

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Abstract
Music has been recognized as an artistic and therapeutic stimulus in humans and other animals. In rodents, most auditory enrichment studies have relied on human-composed music within the human auditory range, overlooking species-specific acoustic perception. Mice communicate mainly in the ultrasonic range (>20 kHz), particularly during courtship, producing structured songs with complex syllables and phrases. To address this gap, we developed and tested a digital ultrasonic music program inspired by murine courtship songs using vocalizations from BALB/c and C57BL/6 strains. Songs were recorded, behaviorally contextualized, and digitally adapted into structured ultrasonic stimuli. Enrichment did not induce anxiogenic effects. In BALB/c mice, ethograms and plus-maze testing revealed increased exploratory behavior in stimulated groups, while C57BL/6 mice showed more variable responses. No adverse effects were observed. This study provides a systematic approach to species-tailored ultrasonic music, generating both behavioral insights and a bioacoustic library that may support replicable protocols in animal enrichment, neuroscience, and welfare research.

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