The radiation emitted from electromagnetic fields (EMF) can cause biological effects on prokaryotic and eukaryotic cells, including non-thermal effects.The present study evaluated the non-thermal effects of wireless fidelity (Wi-Fi) operating at 2.4 GHz part of non-ionizing EMF on different pathogenic bacterial strains ( 0157H7, ). Antibiotic resistance, motility, metabolic activity and biofilm formation were examined.In this case-control, a Wi-Fi router was used as a source of microwaves and also bacterial cells were exposed to Wi-Fi radiation continuously for 24 and 48 hours. The antibiotic susceptibility was carried out using a disc diffusion method on Müller Hinton agar plates. Motility of 0157H7 was conducted on motility agar plates. Cell metabolic activity and biofilm formation were performed using 3-(4, 5-Dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and crystal violet quantification, respectively.The exposure to Wi-Fi radiation altered motility and antibiotic susceptibility of 0157H7. However, there was no effect Wi-Fi radiation on antibiotic susceptibility of . On the other hand, the exposed cells, as compared to the unexposed control, showed an increased metabolic activity and biofilm formation ability in 0157H7, .These results proposed that Wi-Fi exposure acted on bacteria in stressful manner by increasing antibiotic resistance and motility of 0157H7, as well as enhancing biofilm formation by 0157H7, . The findings may have implications for the management of serious diseases caused by these infectious bacteria.