The aim of this research was to assess interactions between metals at low exposure concentrations (Maximum-Permissible-Concentrations accepted for the inland waters in EU) and to assess possible influence of background exposure (10-times reduced concentration of a single metal) on toxicological significance of selected biomarkers in Salmo salar after treatment with metal mixture (Zn - 0.1, Cu - 0.01, Ni - 0.01, Cr - 0.01, Pb - 0.005 and Cd - 0.005 mg/L). The tissue-specific bioaccumulation, genotoxicity and cytotoxicity responses (erythrocytic nuclear abnormalities assay) in peripheral blood, kidneys, gills and liver erythrocytes of fish to metal mixtures were assessed after 14 days treatment. Treatment with primary mixture (MIX) or two variants of this mixture (Cr↓ (10 times reduced Cr concentration) and Cu↓ (10 times reduced Cu concentration)) induced the strongest responses in genotoxicity and cytotoxicity endpoints. Exposure to these mixtures highly affected Zn, Cu and Cd bioaccumulation in liver tissue. The highest amount of Ni accumulated was measured after Cd↓ treatment in all tissues. Treatments with reduced concentration of non-essential metal resulted in an increased accumulation of Pb, Ni, or Cd; treatments with reduced concentration of essential metal resulted in a reduced accumulation of certain metals (especially Cd and Pb) in tissues compared between treatments. Glucose content in blood and behavioural endpoints were evaluated after short-term exposure to metal mixtures (MIX, Cr↓, Cu↓). Significant increase in blood glucose concentration was measured after all treatments. These metal mixtures elicit significant behavioural alterations in fish. Consequently, this research revealed a significant influence of background exposure considering mixture toxicity.

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In the present study, the authors investigated the effect of acclimation duration (up to 4 h) on behavioural characteristics of taxonomically and functionally different fish species, i.e., the migratory rheophilic salmonids rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar), and the non-migratory eurytopic European perch (Perca fluviatilis) and three-spined stickleback (Gasterosteus aculeatus). Specifically, the authors explored fish behavioural patterns based on specific endpoints (average, maximum and angular velocity) during the acclimation period, and determined the acclimation period suitable for the tested fish species. The performed behavioural data analysis showed that the minimum time needed to adjust fish activity to a more stable (baseline) level should be at least 2 h for O. mykiss and S. salar and 1 h for G. aculeatus. Nonetheless, P. fluviatilis behaviour did not show significant changes during the 4 h acclimation. The results of this study revealed that the effect of the acclimation duration on such rheophilic species as O. mykiss and S. salar was greater than that on the eurytopic species P. fluviatilis and G. aculeatus, indicating that acclimation period is important in managing fish stress before behavioural observations. For all species, the highest variability was found in the endpoint of maximum velocity, and the lowest in that of angular velocity. This study showed that before starting actual toxicity testing experiments, it is important to determine an appropriate, species-specific acclimation period.

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