We studied the effects of elevated heavy metal uptake on the sperm quality and the antioxidant mechanisms of sperm and testis of red deer from a Pb mining area in Spain. Testis, liver and bone of red deer from mining (n = 21) and control (n = 20) areas were obtained from hunters and analyzed for Pb, Zn, Cu, Cd, As and Se. Testes were weighed and measured. Motility, acrosome integrity and viability and functionality of membrane were evaluated in epididymal spermatozoa. Lipid peroxidation, total glutathione, glutathione peroxidase (GPX) and superoxide dismutase (SOD) were studied in testis and spermatozoa. Deer from mined areas showed less Cu in testis, a higher testis mass and size and reduced spermatozoa membrane viability and acrosome integrity. Effects on sperm quality were associated to decreased Cu and increased Se in testis, and to decreases in the activity of SOD and GPX in testis and spermatozoa.