Evaluation of genotoxic potential in the Velika Morava River Basin in vitro and in situ Jovana Jovanović a , Stoimir Kolarević a, ⁎, Aleksandra Milošković b , Nataša Radojković b , Vladica Simić b , Biljana Dojčinović c , Margareta Kračun-Kolarević d , Momir Paunović d , Jovana Kostić e , Karolina Sunjog e , Jovana Timilijić a , Jelena Djordjević a , Zoran Gačić e , Bojana Žegura f , Branka Vuković-Gačić a a University of Belgrade, Faculty of Biology, Chair of Microbiology, Center for Genotoxicology and Ecogenotoxicology, Studentski trg 16, 11000 Belgrade, Serbia b University of Kragujevac, Faculty of Science, Institute of Biology and Ecology, Radoja Domanovića 12, 34000 Kragujevac, Serbia c University of Belgrade, Institute of Chemistry, Technology and Metallurgy, Njegoševa 12, 11000 Belgrade, Serbia d University of Belgrade, Institute for Biological Research “Siniša Stanković”, Despota Stefana 142, 11000 Belgrade, Serbia e University of Belgrade, Institute for Multidisciplinary Research, Kneza Višeslava 1, 11000 Belgrade, Serbia f National Institute of Biology, Department of Genetic Toxicology and Cancer Biology, Večna pot 111, SI-1000 Ljubljana, Slovenia HIGHLIGHTS • The study was carried out at 8 sites in the Velika Morava River Basin (VMRB). • Genotoxic potential was assessed in parallel by in vitro and in situ approach. • In vitro assays on native water samples had low potency in the site differentia- tion. • Among in situ assays in bleak blood, alkaline comet assay was the most sensitive. • Integrated biomarker response con- firmed presence of pollution pressure in the VRMB. GRAPHICAL ABSTRACT abstract article info Article history: Received 30 July 2017 Received in revised form 10 October 2017 Accepted 11 October 2017 Available online xxxx The Velika Morava River is the greatest national Serbian river and the significant tributary of the Danube River. The major problems in the Velika Morava River Basin (VMRB) represent untreated industrial and municipal wastewaters. In this study, the level of genotoxic potential at the sites along the VMRB was evaluated by parallel in vitro and in situ approach. Within in vitro testing, genotoxicity of native water samples collected from the sites in VMRB was evaluated by SOS/umuC test on Salmonella typhimurium TA1535/pSK1002 and by the comet assay on HepG2 cells. DNA damage in situ was assessed in bleak (Alburnus alburnus) erythrocytes by the comet (alkaline and Fpg-modified comet) and micronucleus assays. Additionally, the concentration of heavy metals in fish tissue was measured and this data, compiled with the data of the physico-chemical parameters measured in water, was used as a measure of the pollution pressure at the sites. Results showed that applied in vitro tests with native water samples are less sensitive in comparison with in situ tests and should be taken with precaution when making predictions on the status of the ecosystem. Within ap- plied battery of in situ assays differential sensitivity of assays was observed where alkaline comet assay showed the highest potential in differentiation of the sites based on genotoxic potential. Integrated biomarker response showed that usage of the battery of bioassays provides better insight in a genotoxic effects in animals, and consequently, that the holistic approach is more suitable for this type of study. © 2017 Elsevier B.V. All rights reserved. Keywords: Eco/genotoxicity SOS/umuC HepG2 cells Comet assay The Velika Morava River Alburnus alburnus Science of the Total Environment xxx (2017) xxx–xxx ⁎ Corresponding author. E-mail address: stoimirk@bio.bg.ac.rs (S. Kolarević). STOTEN-24323; No of Pages 11 https://doi.org/10.1016/j.scitotenv.2017.10.099 0048-9697/© 2017 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv Please cite this article as: Jovanović, J., et al., Evaluation of genotoxic potential in the Velika Morava River Basin in vitro and in situ, Sci Total Environ (2017), https://doi.org/10.1016/j.scitotenv.2017.10.099