Over the last two decades mobile phone usage has become an essential part of our everyday lives, with a worldwide estimate exceeding one billion users. Although the effects of radiofrequency electromagnetic radiation (RF-EMR) emitted by mobile phones on human health and biological systems are under active debate, several studies have demonstrated that this factor can induce a variety of stresses upon biological systems, including generation of reactive oxygen species and DNA damage. Due to the common practice of storing mobile phone devices in the pant pocket, within close proximity to the reproductive system, it is imperative to detail the effects of RF-EMR on the male germ line. Therefore, in this study C57BL/6 mice were exposed to RF-EMR generated by a waveguide (~2 W/kg intensity, 905 MHz frequency; 12 h a day, for 1-5 weeks) to gain insight into a potential mechanism for RF-EMR associated stress. Mouse spermatozoa were subsequently isolated from the cauda epididymis to assess the effects of RF-EMR treatment. The motility and viability of the spermatozoa proved particularly sensitive to RF-EMR, with significant reductions (p < 0.05) recorded across the categories of rapid, progressive and total motility after 5 weeks, and viability at 1, 3 and 5 weeks. Meanwhile, when spermatozoa were assayed for DNA integrity, it was revealed that levels of oxidative DNA damage and DNA fragmentation were significantly elevated (p < 0.05) in mice receiving RF-EMR treatment. Our continuing research will focus on determining downstream effects of RF-EMR on sperm fertilising ability, and the potential consequences of the acquired DNA damage to the embryo.