The aim of the present study was to ascertain whether the metallothioneins (MTs) of freshwater mussels can serve as a biomarker of environmental quality in the spontaneously polluted area. Anodonta anatina mussels from an agricultural site (A), the cooling pond of a nuclear power plant (N) and a forestry as a reference site (F) in Western Ukraine were studied during spring, summer and autumn. Concentrations of copper (Cu), zinc (Zn) and cadmium (Cd) in MTs and in the tissue, MTs protein (MT-SH) level, oxidative stress indices and apoptotic activities were determined in the digestive gland. The three sites were clear distinguished by biomarkers. In the A-group, the specimens, despite higher concentrations of Cu and Zn in the tissue, had low level of MT-SH level (in two seasons) making the metal-keeping function of MTs invalid. MTs-bound metals dropped to about 2.3% of Cu, 0.4% of Zn and 9.1% of Cd in the tissue in autumn in this polluted area. Lower level of glutathione redox-index reflected the oxidative stress in this group. The N-group displayed high levels of MT-bound Cu (corresponding to up to 57.7% of the total tissue metal) and caspase-3 activity (up to 7 times higher than in the reference Fgroup), indicating specific for cooling pond pollution by Cu and thermal effects. In group F, the set of markers showed the characteristic of the reference site. An extremely low redox-index of glutathione (RI GSH: 0.45) was observed in the A-group in spring, and high levels of protein carbonyls and TBA-reactive substances in autumn in group N.Hence, multi-marker approach needed to provetoxicityassesments in mussel sexposed byspontaneous and mixed pollution.
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