Copper is a transition metal that exists in different chemical forms (e.g., Cu2+,Cu+, and Cu-0) and at high concentrations it is toxic. Here, we investigated the Cu2+-induced toxicity in Drosophila melanogaster, evaluating the survival, locomotion, and the activity of acetylcholinesterase (AChE) and glutathione S-transferase (GST) enzymes. Flies were exposed to Cu2+(0.1-1mmol CuSO4/kg of diet or approximately 0.1-1mM Cu2+) and allowed to mate during 24h. GST and AChE enzymes were evaluated in the larvae and in the head and the body (thorax + abdomen) of the adult male and females flies. The total number of adult females (0.4-1mM) and males (0.75 and 1mM) was decreased by CuSO4. The climbing ability was hampered in flies exposed to 1mM Cu2+. In larvae, Cu2+(0.4-1mM) increased AChE activity (P<0.002). In males' heads, 0.4mM Cu2+ increased the AChE activity (P<0.01). In adults' bodies, Cu(2+)inhibited the activity in both sexes, but with greater effectiveness in males (0.1 to 1mM) than in females (1mM). Regarding GST activity, 0.1mM Cu(2+)increased, but 1mM decrease GST in larvae. In the head of flies, Cu(2+)decreased the GST activity at intermediate (0.4mM) and increased GST at the highest concentration (1mM) in males. In the bodies, the effect of Cu(2+)was similar. In conclusion, Cu(2+)exposure in D. melanogaster disrupted locomotion and enzymatic parameters that can be related to changes in AChE and in the detoxifying GST enzyme.