Background: The aim of this study was to investigate the effect of the electromagnetic field (EMF) generated by 10 kV (50 Hz) high voltage line on the epididymal sperm characteristics and the protective effects of ganoderma lucidum (GL) and melatonin (M) in EMF exposed male rats. Materials and Methods: In this study, 64 Wistar rats were divided into 8 groups (n= 8). Rats in 26-day and 52-day experiment groups were exposed to EMF for 8 hours daily during 26 and 52 days respectively. Rats in EMF+GL-26 and EMF+GL-52 groups were treated with GL by oral gavage 20 mg/kg daily and rats in EMF+M-26 and EMF+M-52 groups received M 10 mg/kg daily during experiments. At the end of experiments, all rats were sacrificed and epididymal sperm concentration, motility and morphology were evaluated. Results: In the 26-day experiment, sperm motility of the control group were significantly lower than all groups (P<0.05). In the 52-day experiment, the EMF+GL52 group had the lowest tail and total morphologic defect rates and the differences between this group and EMF52 and EMF+M52 groups were significant. Conclusion: Our data demonstrated that; (1) EMF exposure can have different effects on sperm quality according to duration of exposure, (2) treatment of GL in rats exposed to EMF for 26 or 52 days increased sperm concentration and reversed negative effect of EMF on sperm morphology, respectively and (3) treatment of M in rats exposed to EMF for 26 or 52 days had negative effects on sperm concentration and sperm morphology, respectively.