Although amphibians are considered to be good bioindicators of environmental pollution, few data are available concerning their biochemical parameters in natural populations. We investigated seasonal (spring, summer, autumn) and spatial (wetlands in rural and urban areas) fluctuations of oxidative stress biomarkers in the liver of frog Rana ridibunda in Western Ukraine. The Centroid grouping analysis demonstrated that despite the fluctuations of separate indices, frogs from an urban site in summer and autumn are differed widely from those at the same site in spring and frogs from the rural site in all three seasons, joined in common set. In summer, suppression of Mn-superoxide dismutase and catalase activity, as well as increase in oxidized glutathione and lipid peroxidation levels demonstrate a collapse of antioxidant defense system in frogs from an urban site. The integrated oxidative stress index confirms this conclusion.