The goal of this study is to investigate the relationships between total length and otolith dimensions and detect the otolith shape index values of Rhodeus amarus sampled from the two different localities (Terme and Terice Streams) in Turkey. A total of 116 specimens of R. amarus were collected. Power models were applied to determine the relationships between otolith dimensions and fish length. Otolith indices were calculated for asteriscus and lapillus pairs. R. amarus were studied ranged in total length from 4.5–8.9 cm for two localities. When the relationships between total length, otolith length and otolith width were examined, the best fit was obtained among total length and otolith length in two localities for both asteriscus and lapillus (r ² > 0.705). Analysis for otolith measurements (asteriscus and lapillus) clearly demonstrated that there was a difference in terms of otolith lengths between samples of Terme and Terice Streams. However, form factor, circularity, roundness values of asteriscus and rectangularity, aspect ratio, ellipticity of lapillus are different between Terme and Terice Streams (p < 0.001).