Metals can cause oxidative stress by increasing the formation of reactive oxygen species (ROS), when there are insufficient amount of antioxidants to defend against the growing amount of free radicals. We aimed to find out the most reliable biomarkers to detect pollution-related oxidative stress in wild birds by comparing oxidative stress status in great tit (Parus major) nestlings at populations in polluted and unpolluted areas. We also studied with experimental manipulations whether dietary carotenoid levels have any role in great tits' antioxidant defence and whether their carotenoid-based plumage colour was connected to an oxidative stress status. We used antioxidants (GSH, carotenoids) and several antioxidant enzymes (GP, GR, GST, SOD, and CAT) as indicators of the oxidative stress. We found no direct connections between dietary metal exposure and antioxidant or antioxidant enzyme levels. The activity of GP was, however, slightly higher in the polluted environment. This was due to poorer condition and subsequently higher level of oxidative stress in the nestlings in the polluted area. We also found a positive association between GP and an ambient temperature during the nestling period, which may be due to higher metabolic activity of partly poikilothermic nestlings in warm weather. The activity of GST was positively related to the number of nestlings at the sampling time. Fledging success was better in an unpolluted area, where also the nestling body mass was higher. Carotenoid treatment increased the plasma carotenoid concentrations 2.1 fold in carotenoid-supplemented birds, but was not associated with the oxidative stress biomarkers or metal levels. The yellow plumage colour was associated with dietary carotenoid levels in both study areas, but not with the metal exposure or the oxidative stress status. Our results suggest that at the exposure levels found in our study area, the enzyme activities do not indicate metal-related oxidative stress. Instead, GP can be used as an indicator of growth related oxidative stress, which is greater in the polluted area. The activity of this enzyme was, however, not directly related to metal exposure, but more likely to some secondary pollution-related change in the nestling condition.