Oxidative status in relation to metal pollution and calcium availability in pied flycatcher nestlings – A calcium manipulation experiment

Metal exposure can produce oxidative stress by disrupting the prooxidant/antioxidant balance. It has been suggested that calcium (Ca) may provide protection against metal toxicity in the organism. The objective of this study is to explore the effects of Ca availability and metal pollution on oxidative stress biomarkers in pied flycatcher (Ficedula hypoleuca) nestlings. For this purpose, we performed a Ca-supplementation experiment with birds inhabiting a Ca-poor and metal-polluted area in SW Finland. An array of oxidative stress biomarkers (GSH, GSH:GSSG ratio, GPx, GST, CAT, SOD, lipid peroxidation and protein carbonylation) was measured in red blood cells. The effects of antioxidant molecules and oxidative damage on nestling size, growth, fledging success and fledgling number were evaluated. We observed an up-regulation of GST activity and increased protein carbonyl content in the polluted zone, probably related to a combination of higher metal exposure and reduced food quantity and quality in this area. As expected, birds from the unpolluted zone showed higher GSH:GSSG ratio but, unexpectedly, also showed signs of higher lipid peroxidation (not statistically significant, p = 0.056), both responses likely being related with the lower Ca availability. Our study suggests that different measures of oxidative damage are affected by different factors: while damage to proteins was the target of metal exposure/food limitation, poor Ca availability may enhance damage to lipids in growing birds. The intercorrelations found between Ca in plasma, metal exposure and the different oxidative stress biomarkers show that the antioxidant defense is finely regulated to cope with increased oxidative challenges. Finally, our results suggest that the antioxidant status during early development, conditioned by environmental pollution and Ca availability, is one factor affecting nestling survival and fledgling number.