The purpose of this study was to investigate the effects of polluted Soummam River and unpolluted Agrioun River on sperm parameters and oxidative stress status of Barbus callensis spermatozoa during the spawning season in natural condition. The experimental design consisted to activate alternatively fish sperm of the two sites with the polluted (Soummam River, S) and unpolluted water (Agrioun River, A). Sperm motility duration (SMD) was measured using a stopwatch. Gametes straight line velocity (VSL), average path velocity (VAP), curvilinear velocity (VCL), spermatozoa concentration (SC), straightness (STR) and linearity (LIN) were measured by a CASA. Oxidative stress biomarkers were evaluated by measuring total antioxidant status (TAS) and catalase (CAT) activity. The results showed that the SMD and spermatozoa velocity were significantly higher in (Sm, S) than in (Ag, A) with SMD = 52 versus 42s, VSL = 23 versus 16 µm/s, VAP = 35 versus 25 µm/s, and VCL = 52 versus 35 µm/s, respectively. However, SC, STR and LIN were significantly higher in (Ag, A) than in (Sm, S) with SC = 37.5x109 versus 27x109 spz/ml, STR = 52 versus 40% and LIN = 35 versus 26%. Likewise, the oxidative status of fish spermatozoa was significantly affected by the quality activating water; TAS and CAT were significantly higher in (Ag, A) than in (Sm, S); 7.5 to 0.5 and 120 to 28 µmol/min/ml, respectively. The current investigation showed that Barbus callensis sperm motility parameters, particularly spermatozoa concentration, straightness and linearity are good bioindicators of water pollution.