Quantifying the response of grass carp larvae to acoustic stimuli using particle-tracking velocimetry

Acoustic deterrents are recognized as a promising method to prevent the spread of invasive grass carp, Ctenopharyngodon idella (Valenciennes, 1844) and the negative ecological impacts caused by them. As the efficacy of sound barriers depends on the hearing capabilities of carp, it is important to identify whether carps can recognize acoustic signals and alter their swimming behavior. Our study focuses on quantifying the response of grass carp larvae when exposed to out-of-water acoustic signals within the range of 100-1000 Hz, by capturing their movement using particle-tracking velocimetry (PTV), a quantitative imaging tool often used for hydrodynamic studies. The number of responsive larvae is counted to compute response ratio at each frequency, to quantify the influence of sound on larval behavior. While the highest response occurred at 700 Hz, we did not observe any clear functional relation between frequency of sound and response ratio. Overall, 20-30% of larvae were consistently reacting to sound stimuli regardless of the frequency. In this study, we emphasize that larval behaviors when exposed to acoustic signals vary by individual, and thus a sufficient number of larvae should be surveyed at the same time under identical conditions, to better quantify their sensitivity to sound rather than repeating the experiment with individual specimens. Since bulk quantification, such as mean or quantile velocities of multiple specimens, can misrepresent larval behavior, our study finds that including the response ratio can more effectively reflect the larval response.