The aim of this study was to determine the optimal levels of Na+, Ca+2, k+, Sucrose and osmotic pressure in saline activator solutions for increasing the duration of spermatozoa motility in Barbus sharpeyi. Also, the effect of different saline activator solutions were assessed on VCL, VSL and Linearity of spermatozoa cells in B. sharpeyi. Results showed that Na+ ion concentration of 100 mML-1, 50 mML-1 of k+ and 2.5 mML-1 of Ca+2 have created the optimal conditions for stability of spermatozoa movement. The optimal level of osmotic pressure in sperm activator solution was 179±3.4 mOsmol kg-1. Results showed that the highest VCL and VSL were assessed by 50 mM NaCl. Also, 100 mML-1 of Na+ and 20 mML-1 of Tris induced the highest Linearity percentage in B. sharpeyi spermatozoa (p≤0.05). In final conclusion, usage of extender solutions containing Na+ ion concentration of 100 mML-1, 50 mML-1 of k+ , 2.5 mML-1 of Ca+2 and 150 mML-1 sucrose is suggested separately for obtaining the optimal motility of spermatozoa in Barbus sharpeyi. Also, the mixed extender solution containing concentration 50 mML-1 of NaCl, 30 mML-1 of KCl, 2.5 mML-1 of Ca+2, 30 mML-1 of Tris and 189 mOsmol kg-1 of osmotic pressure is suggested for activation of spermatozoa motility in Barbus sharpeyi.