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Combined use of biomarkers to assess the impact of untreated wastewater from the Danube River, Serbia

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Abstract

In this study a battery of bioassays, both in vivo (metals and metalloids concentrations, erythrocyte morphometry, comet assay, micronucleus assay, and histopathological analyses) on vimba bream Vimba vimba (L., 1758) and white bream Blicca bjoerkna (L., 1758), and in vitro (treatment of HepG2 cells with native water samples) was applied to assess the harmful potential of untreated wastewater. Faecal indicator bacteria were quantified to assess the microbiological water quality. Vimba bream had significantly higher Fe concentrations in both liver and muscle, while white bream had higher Ca and Cu concentrations in liver. Vimba bream had a significantly higher level of DNA damage in both liver and blood cells, in comparison to white bream. Low levels of micronucleus and nuclear abnormalities were observed in both species. Erythrocytes morphometry did not show significant interspecific differences. Histopathological analyses revealed a similar response of the studied species, with a significantly higher presence of ceroid pigments in the liver of vimba bream. Treatment of HepG2 cells revealed the high genotoxic potential of water downstream of the discharge point. The results of this study clearly demonstrate the importance of effect-based monitoring, in order to enforce more efficient management of natural resources and implementation of wastewater treatment systems.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This research was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Contract Nos. 451-03-68/2022-14/200053, 451-03-68/2022-14/ 200178, 451-03-68/2022-14/200007, 451-03-68/2022-14/200116). We gratefully acknowledge technician Zorica Radović for preparing the histological samples. The improvement of our English by Luka Gačić is greatly appreciated.

Author contributions

ML and BV-G contributed to the study design, conceptualisation, and resources. JK-V and SK contributed to material preparation, data collection, and analysis of the comet assay on HepG2 and fish cells, as well as microbiological analysis. KS performed a micronucleus assay. SS conducted a study of erythrocyte morphometry. ŽV-J performed the analysis of metals and metalloids concentrations in fish tissues. VP and BR were responsible for the histopathological analyses of fish tissues. The first draft of the manuscript was written by JK-V. All authors commented on previous versions. All authors read and approved the final manuscript.

Funding

This research was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Contract Nos. 451-03-68/2022-14/200053, 451-03-68/2022-14/200178, 451-03-68/2022-14/200007, 451-03-68/2022-14/200116).

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Authors and Affiliations

  1. University of Belgrade, Institute for Multidisciplinary Research, Department of Biology and Inland Waters Protection, Belgrade, Serbia

    Jovana Kostić-Vuković, Karolina Sunjog, Željka Višnjić-Jeftić & Mirjana Lenhardt

  2. University of Belgrade, Institute for Biological Research “Siniša Stanković”, National Institute of Republic of Serbia, Department of Hydroecology and Water Protection, Belgrade, Serbia

    Stoimir Kolarević & Mirjana Lenhardt

  3. University of Belgrade, Faculty of Biology, Chair of Animal Ecology and Zoogeography, Belgrade, Serbia

    Srđan Subotić

  4. University of Belgrade, Faculty of Agriculture, Institute of Animal Science, Belgrade, Serbia

    Božidar Rašković & Vesna Poleksić

  5. University of Porto, Institute of Biomedical Sciences Abel Salazar (ICBAS), R. Jorge de Viterbo Ferreira 228, 4050-313, Porto, Portugal

    Božidar Rašković

  6. University of Belgrade, Faculty of Biology, Chair of Microbiology, Center for Genotoxicology and Ecogenotoxicology, Belgrade, Serbia

    Branka Vuković-Gačić

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  1. Jovana Kostić-Vuković
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Correspondence to Jovana Kostić-Vuković.

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Kostić-Vuković, J., Kolarević, S., Sunjog, K. et al. Combined use of biomarkers to assess the impact of untreated wastewater from the Danube River, Serbia. Ecotoxicology 32, 583–597 (2023). https://doi.org/10.1007/s10646-023-02663-6

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