The electric fields (EFs) generated by high-voltage power lines used to transmit electricity among cities and villages may have the considerable potential to significantly influence organisms in close proximity. This study aims to investigate alternations in Drosophila melanogaster biology and behavior following exposure to high EFs. Moreover, the levels of neurochemicals in flies exposed to EF were evaluated. A simulation system program was used in the laboratory to generate actual EF values in the vicinity of high-voltage power lines. The intensity of EF was adjusted to 12.0 kV/m and 5.7 kV/m, representing two different distances from a 220 kV power line. Flies were exposed to the simulated EFs for 6 days/8 hours a day. The results revealed that near-distance exposure to EFs negatively impacted development time, adult emergence, and hatchability rate. In behavior, EF exerted profound adverse effects on memory retention, climbing ability, male aggression, and adult food consumption. Additionally, near-distance exposure to EFs showed a significant increase in the levels of neurotransmitters, dopamine and serotonin. The results provide scientific evidence that anthropogenic EFs emitted from the transmission power lines in terrestrial habitats can be an environmental stressor potentially affecting the biology, behaviour, and neurochemicals of Drosophila. We assume a similar impact of these high EFs on vertebrates including humans living near high-voltage power lines. Therefore, the study recommends avoiding building under high-voltage lines, and people should reside far away from these lines when constructing any facility for humans or living organisms.