A quantitative and comparative analysis of the acoustic whistles parameters of tucuxi dolphins (Sotalia fluviatilis) from eight areas in Brazilian waters were conducted. Tucuxi produced mainly upsweep whistles, and despite their similar characteristics to other delphinids, whistles were shorter and less complex in shape. Some signals had fundamental frequency components above 24 kHz, and a broader band system may be useful in further studies concerning tucuxi whistles. The frequency parameters of the whistles had the lowest variation coefficients, whereas the duration and number of inflections had the highest variation coefficients. About 65% of all whistles had common characteristics between study localities, but SF, MinF, MeF, F1/4, F1/2, and F3/4 increased in frequency from southern to northern populations. The magnitude of the whistle characteristics variation was comparatively smaller between adjacent sites areas than between nonadjacent ones. However, for some pairwise comparisons the whistles were more similar between nonadjacent areas than between adjacent ones, indicating that other factors besides genetic and mixing affected tucuxi whistle characteristics. Only a few numbers of whistle variables were significantly different between adjacent areas and the parameter ranges of whistles overlapped, which made it difficult to discriminate whistles between adjacent tucuxi populations.