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The strong negative anthropogenic pressure on the River Bregalnica resulted in the increase of pollution in the river ecosystem. The aim of this study was to evaluate if aquatic pollutants generate genotoxic effects in barbel (Barbus peloponnesius). Thirty individuals were collected from three localities along the river: one control site and two locations in the middle and lower course of the river with the presence of different pollution sources. Blood smears were stained with May-Grünwald/Giemsa method and the frequency of micronuclei, vacuolated nuclei, binuclei, blebbed nuclei and notched nuclei expressed in per mill was recorded. In fish collected from the control site, the investigated erythrocyte nuclear abnormalities revealed lowest values compared to polluted sites. In general, significantly higher values for erythrocyte deformations, including micronuclei, were noted in the middle and lower part of the river. Significantly higher values of vacuolated and blebbed nuclei were detected in fish from the middle course of the river, while at the lower course the frequencies of occurrence of micronuclei and notched nuclei were significant compared with control site. The middle course of the River Bregalnica has more diverse pollutants compared to the lower course, and consequently the highest frequency of all nuclear deformations was found in barbel from the middle course of the river. Obtained results suggest that pollutants draining into the Bregalnica river ecosystem display a genotoxic effect on the local barbel population.

Freshwater contamination with mining waste can result with high concentrations of toxic metals in the water and in fish organs. In North-Eastern Macedonia, several rivers (e.g., Zletovska, Kriva) are exposed to acid mine drainage from active Pb/Zn mines. Previous studies confirmed high concentrations of dissolved metals in their water. This study was performed in liver and gills of Vardar chub (Squalius vardarensis Karaman, 1928) from three Macedonian rivers (Bregalnica, Kriva and Zletovska) in spring and autumn 2012. The aim was to establish if increased exposure to certain metals have resulted with their increased bioaccumulation. The concentrations of 19 elements were measured in cytosolic tissue fractions, to obtain information on metabolically available metal species. The following ranges of cytosolic concentrations of highly toxic elements were measured in the Vardar chub liver (in μg/L): Cd, 1.18–184; Cs, 0.25–25.4; Tl, 0.02–5.80; Pb, 0.70–61.1. Their ranges measured in the gills (in μg/L) were the following: Cd, 0.24–59.2; Cs, 0.39–24.4; Tl, 0.01–1.00; Pb, 0.65–87.2. Although the water of the mining impacted Zletovska River was highly contaminated with several essential metals, especially with Mn and Zn, the majority of essential elements (Na, K, Ca, Mg, Co, Cu, Fe, Mn, Mo, and Zn) did not reflect the exposure level. In contrast, seven nonessential elements reflected the level of exposure in the water. Significantly increased hepatic and gill concentrations of Cs, Rb, Sr, and Tl were detected in Vardar chub from the Zletovska River compared to the other two rivers, of Cd and Pb in the Zletovska and Kriva River compared to Bregalnica, and of V in the Bregalnica River compared to Zletovska and Kriva rivers. Observed significant metal bioaccumulation, in particular of highly toxic elements, as a consequence of exposure to water contaminated with mining waste points to necessity of intensified supervision of mining impacted rivers.