Abstract: | The aim of this study was to evaluate the effects of osmolality and the presence of ions on the activation of post‐thaw sperm motility of Brycon insignis. Sperm was frozen under a standardized methodology for this species. In experiment 1, 11 solutions were prepared with reverse osmosis (RO) water (~0 mOsm/kg) and glucose or NaCl adjusted to an osmolality of 50, 100, 150, 200 and 250 mOsm/kg. In experiment 2, six solutions were prepared with RO and adjusted to ~100 mOsm/kg with one of the following chemicals: NaHCO3, sodium citrate (Na3C6H5O7), NaCl, KCl, CaCl2 or glucose (as ion‐free control). Post‐thaw sperm of both experiments was evaluated for motility rate, velocities (curvilinear = VCL, among others) and beat‐cross frequency (BCF). In experiment 1, sperm motility rate and velocities were higher (p < 0.05) when triggered in solutions at osmolalities from 0 to 200 mOsm/kg (62–80% motility; 139–167 µm/s) than that at 250 mOsm/kg (36–44% motility; 94–99 µm/s VCL). BCF was not affected by osmolality and varied from 19 to 24 Hz in all samples. In experiment 2, samples activated in NaHCO3, citrate, NaCl and KCl solutions yielded higher motility rates (76–85%) and BCF (24–25 Hz) compared to those activated in CaCl2 (50%; 14 Hz). Samples activated in ion‐free control solution yielded higher motility rate (87%) than those activated in NaHCO3 and in CaCl2. Curvilinear velocity was higher in samples activated in NaHCO3, citrate, KCl and control solutions (144–160 µm/s) than in those activated in CaCl2 (104 µm/s); samples activated in NaCl yielded intermediate VCL values (127 µm/s). Post‐thaw sperm achieves maximum motility rate and velocities when activated in solutions composed of sodium citrate, NaCl, KCl or glucose. Thus, post‐thaw sperm motility of B. insignis can be triggered in ionic and non‐ionic solutions at osmolality between 0 and 200 mOsm/kg. The use of solutions containing calcium, however, should be avoided. |