The target of this in vitro study was to investigate the dose- and time-dependent effects of copper (CuSO4.5H2O) in different culture media on the spermatozoa motility and to provide additional information on the interaction between serum albumin and copper ions. The spermatozoa motility was determined after exposure of CuSO4.5H2O (3.90; 7.80; 15.60; 31.20; 62.50; 125; 250; 500; 1000 µmol.dm-3) using the SpermVisionTM CASA system (Computer Assisted Semen Analyzer) during different time periods (Time 0 h, 1 h, 2 h, 24 h). The initial percentage of motility spermatozoa in the presence CuSO4.5H2O in physiological saline solution (PS) showed significantly (P<0.001) decreased values at high concentrations ≥ 250 µmol.dm-3 of CuSO4.5H2O. The low concentrations (≤ 7.80 µmol.dm-3 of CuSO4.5H2O) maintained of spermatozoa motility (Time 2 h). The long-term cultivation significantly (P<0.001) reduced the average motility values in all experimental groups compared to the control group (medium without CuSO4.5H2O). The commercial medium (CM) containing triladyl, egg yolk and redistilled water increased the overall percentage of spermatozoa motility after exposure of high doses of CuSO4.5H2O, but only during short-time periods. The culture medium in composition of 20.0 % bovine serum albumin (BSA), triladyl, 5.0 % glucose and redistilled water maintained the spermatozoa motility in all experimental groups (Time 0 h, 1 h). Evaluation of the overall percentage of spermatozoa motility showed significant (P<0.001) decrease at high concentrations ≥ 500 µmol.dm-3 of CuSO4.5H2O after 2 h and after 24 h of cultivation when exposed to doses ≥ 125 µmol.dm-3 of CuSO4.5H2O. The obtained data point out that copper at high doses acts as toxic element on the spermatozoa motility parameters. However, using a suitable culture medium containing an energy component- and protein-rich substrate, the spermatozoa motility could increase.