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Comparison of comet assay parameters for estimation of genotoxicity by sum of ranking differences

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Abstract

The genotoxic potential of waters in six rivers and reservoirs from Serbia was monitored in different tissues of chub (Squalius cephalus L. 1758) with the alkaline comet assay. The comet assay, or single-cell gel electrophoresis, has a wide application as a simple and sensitive method for evaluating DNA damage in fish exposed to various xenobiotics in the aquatic environment. Three types of cells, erythrocytes, gill cells, and liver cells, were used for assessing DNA damage. Images of randomly selected cells were analyzed with a Leica fluorescence microscope and image analysis by software (Comet Assay IV Image analysis system, PI, UK). Three parameters (tail length—l, tail intensity—i, and Olive tail moment—m) were analyzed on 1,700 nuclei per cell type. The procedure for sum of ranking differences (SRD) was implemented to compare different types of cells and different parameters for estimation of DNA damage. Regarding our nine different estimations of genotoxicity: tail length, intensity, and moment in erythrocytes (rel, rei, rem), liver cells (rll, rli, rlm), and gill cells (rgl, rgi, rgm), the SRD procedure has shown that the Olive tail moment and tail intensity are (almost) equally good parameters; the SRD value was lower for the tail moment and tail intensity than for tail length in the case of all types of cells. The least reliable parameter was rel; close to the borderline case were rei, rll, and rgl (∼5 % probability of random ranking).

Comparison of comet assay parameters

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Acknowledgments

This study represents a part of the activities within project no. 173045, funded by the Ministry of Education and Science of the Republic of Serbia.

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Authors and Affiliations

  1. Department of Natural Resources and Environmental Sciences, Institute for Multidisciplinary Research, University of Belgrade, Kneza Višeslava 1, 11030, Belgrade, Serbia

    K. Sunjog, Z. Gačić & M. Lenhardt

  2. Center for Genotoxicology and Ecogenotoxicology, Faculty of Biology, University of Belgrade, Studenski Trg 16, 11000, Belgrade, Serbia

    S. Kolarević, J. Knežević-Vukčević & B. Vuković-Gačić

  3. Research Centre for Natural Sciences, Hungarian Academy of Sciences, Pusztaszeri ut 59/67, 1025, Budapest, Hungary

    K. Héberger

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  1. K. Sunjog
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  2. S. Kolarević
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  3. K. Héberger
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  4. Z. Gačić
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  5. J. Knežević-Vukčević
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Correspondence to K. Sunjog.

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Sunjog, K., Kolarević, S., Héberger, K. et al. Comparison of comet assay parameters for estimation of genotoxicity by sum of ranking differences. Anal Bioanal Chem 405, 4879–4885 (2013). https://doi.org/10.1007/s00216-013-6909-y

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  • DOI: https://doi.org/10.1007/s00216-013-6909-y

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