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River water in Kosovo is exposed to various discharges from industrial and agricultural activities as well as to urban wastewater. Rivers Sitnica and Drenica are among the most affected ones and water samples drawn from these rivers show the presence of various toxic substances. Genotoxic effects are seen in fish living in these rivers indicating a cytotoxic and mutagenic potential of the river water. Aiming at substantiating these observations, we assessed the cyto- and genotoxic effects of water samples collected at different locations from the Drenica and Sitnica rivers. Samples drawn from Lake Badovc served for comparison. To address seasonal effects, samples were collected at different seasons/time points during the period of summer 2016-spring 2018. The water samples were analyzed employing primary rat hepatocytes as a reliable in vitro cell model for the assessment of cytotoxic effects (mitotic arrest and cell death) and DNA damage/genotoxicity (micronucleus assay and Comet assay). The results do not account for significant effects associated with specific locations but demonstrate seasonal differences of the genotoxic potential of the water samples collected along both rivers, which are accompanied by a limited cytotoxic potential. Our data provide substantial support to earlier observations and strongly warrant the need for continuous chemical as well as biological monitoring of the river water in Kosovo, focusing on improved toxicant profiling of the river water and investigations addressing the observed seasonal variations., (© The Author(s) 2024. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Being exposed to untreated urban and industrial water, the rivers Drenica and Sitnica are considered to be the most polluted ones in the Kosovo. Our previous investigations on the cyto- and genotoxic potential of water samples from these rivers evaluated with primary rat hepatocyte cultures indicated a risk for the health of aquatic organisms. In order to assess the genotoxic risk to aquatic organisms, we therefore performed a two year study (2016-2017) on roach (Rutilus rutilus) from these rivers. Specimens were collected at three locations along the Drenica river and two locations along the Sitnica river, and the genotoxicity was evaluated by the micronucleus as well as the Comet assay (DNA damage) in erythrocytes. The frequencies of micronucleated cells were determined for samples collected in four seasons, whereas the Comet assay was employed on samples collected in five seasons during the two-year period. The data obtained revealed an increase of the frequency of micronucleated erythrocytes from Rutilus rutilus collected at most sampling locations and from both rivers at all seasons investigated. Significant differences to the control (lake Badovc) were found in summer 2016 and spring 2017 samples. When comparing the seasons, the summer 2016 samples were most genotoxic, followed by spring 2017 and autumn 2016. With regard to the Comet assay data, a similar but more prominent "response" was observed. Another important observation is that micronucleus rates as well as DNA damage levels were significantly higher in samples collected in 2016 compared to the respective seasons in 2017. Altogether, the "response" obtained with both markers confirmed a genotoxic risk for fish due the pollution of these rivers. Since there were, however, seasonal and annual variations of the genotoxicity levels further in depth studies have to be carried out addressing the nature of these changes., (Copyright © 2019. Published by Elsevier B.V.)