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Beauvericin and ochratoxin A genotoxicity evaluated using the alkaline comet assay: single and combined genotoxic action

  • Genotoxicity and Carcinogenicity
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Abstract

This study was aimed at investigating the genotoxic potential of single beauvericin (BEA) and ochratoxin A (OTA) as well as their interaction in porcine kidney epithelial PK15 cells and human leukocytes using the alkaline comet assay. IC50 of BEA (5.0 ± 0.6) and OTA (15.8 ± 1.5) estimated by MTT reduction assay shows that BEA is three times more toxic than OTA. BEA (0.1 and 0.5 μM) and OTA (1 and 5 μM) were applied alone or in combination of these concentrations for 1 and 24 h in PK15 cells and human leukocytes. Genotoxicity of these toxins to PK15 cells was time- and concentration dependent. After 1 h, significant increase in tail length, tail intensity, tail moment, and abnormal sized tails (AST) was noted upon exposure to 1 μM of OTA alone and BEA + OTA combinations. Single BEA (0.5 μM) and OTA (1 and 5 μM) and their combinations evoked significant DNA damage in PK15 cells, considering all comet tail parameters measured after 24 h of treatment. Human leukocytes were slightly concentration but not time dependent. After 1 h of exposure, there were no significant changes in the tail length. Tail intensity, tail moment, and/or incidence of AST were significantly higher in cells treated with single OTA or BEA and their combinations than in control cells. DNA damage in leukocytes was significantly higher after 24 h of exposure to single toxins and their combinations, considering all comet tail parameters, but these changes were less pronounced than in PK15 cells. Combined toxins showed additive and synergistic effects in PK15 cells, while only additive effects were observed in human leukocytes. Combined prolonged exposure to BEA and OTA in subcytotoxic concentrations through food consumption could induce DNA damage contributing to the carcinogenicity in animals and humans.

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Acknowledgments

We wish to thank Professor Josip Madić, PhD and Snjezana Kovač, MSc (Veterinary Faculty, University of Zagreb, Croatia) for providing the PK15 cells. Our thanks are also due to Dado Čakalo for language advice. This work was financially supported by the Ministry of Science, Education, and Sports of the Republic of Croatia (Grants No. 022-0222148-2137, 006-0061117-1242).

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Pharmacy and Biochemistry, University of Zagreb, Schrottova 39, 10000, Zagreb, Croatia

    Maja Šegvić Klarić, Dina Daraboš & Stjepan Pepeljnjak

  2. Mutagenesis Unit, Institute of Medical Research and Occupational Health, Zagreb, Croatia

    Ružica Rozgaj & Vilena Kašuba

Authors
  1. Maja Šegvić Klarić
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  2. Dina Daraboš
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  3. Ružica Rozgaj
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  4. Vilena Kašuba
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  5. Stjepan Pepeljnjak
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Correspondence to Maja Šegvić Klarić.

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Klarić, M.Š., Daraboš, D., Rozgaj, R. et al. Beauvericin and ochratoxin A genotoxicity evaluated using the alkaline comet assay: single and combined genotoxic action. Arch Toxicol 84, 641–650 (2010). https://doi.org/10.1007/s00204-010-0535-7

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  • DOI: https://doi.org/10.1007/s00204-010-0535-7

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