Impact of activation solutions on fresh and frozen-thawed sperm motility and fertilization success for two species of migratory freshwater fishes

Abstract

Extracellular environment conditions, ionic concentration, pH, osmolality, and temperature influence sperm activation and sperm quality. The aim of this study was to evaluate the effects of different activating solutions on sperm quality and fertilization rate of fresh and post-thaw sperm in Brycon orbignyanus and Prochilodus vimboides. Activation solutions with ions: NaCl, KCl, CaCl2 (150 mOsm kg−1), without ions: glucose (150 mOsm kg−1) and water: reverse osmosis (RO) (∼0 mOsm kg−1) and tank water (TW) were tested. In experiment 1, fresh sperm motility was activated in each activating solution and motility rate (%), motility quality score (0–5), and motility duration (seconds) were subjectively evaluated using a microscope. In experiment 2, sperm was cryopreserved and post-thaw sperm quality was assessed in each activating solution. Methyl glycol was used as cryoprotectant and for B. orbignyanus a solution of BTS® 5% in water reverse osmosis - 325 mOsm kg−1 was used as extender, while for P. vimboides it was used a solution of glucose 5% in water reverse osmosis - 325 mOsm kg−1. In straw, cryoprotectant, extender, sperm were respectively 10%, 80% 10% (V/V). B. orbignyanus fresh sperm activated in NaCl, KCl, glucose solutions, TW and RO yielded higher averages for all the subjective parameters analysed. In fresh sperm of P. vimboides the highest values were observed when glucose solution was used for sperm motility activation, and the highest fertilization rates were observed in samples activated in glucose or RO solutions. B. orbignyanus post-thaw sperm activated in TW (45.1%) or RO (39.7%) presented the highest values for motility. The highest values of curvilinear velocity (VCL) were observed using glucose (69.5 μm s−1), NaCl (67 μm s−1) and KCl (68.4 μm s−1), but the highest fertilization rates were observed when glucose (3.6%), RO (3.5%) and TW (2.5%) were used. P. vimboides post-thaw sperm activated in glucose solution presented the highest motility rate (41%), VCL (43.7 μm s−1), fertilization rate (18.2%) and hatching rate (13.7%). In order to achieve the best seminal quality, fertilization and hatching rates in both fresh and post-thaw sperm, the glucose solution, TW and RO are indicated for use as sperm motility activators in B. orbignyanus, whereas for P. vimboides the glucose solution and RO are indicated.