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In vitro study of mutagenicity of cylindrospermopsin and cytotoxic effects of the combination of microcystin and cylindrospermopsin
Abstracts / Toxicology Letters 258S (2016) S62–S324
P09-013 In vitro study of mutagenicity of cylindrospermopsin and cytotoxic effects of the combination of microcystin and cylindrospermopsin A. Jos ∗ , D. Gutiérrez Praena, S. Maisanaba, M. Llana Ruíz Cabello, A. Cameán Area of Toxicology, Faculty of Pharmacy, University of Sevilla. C/Profesor García González 2, 41012 Sevilla, Spain Cylindrospermopsin (CYN) and microcystin-LR (MC-LR) are two cyanotoxins produced by several species of cyanobacteria and they are well distributed worldwide. It is known these toxins, separately, induce harmful effects in human, animals and plants, included those destined for human consumption. However, some of their toxic effects are still scarcely investigated, such as the mutagenicity induced by CYN. Moreover, CYN and MC-LR can appear in water reservoirs at the same time, so it could be assumed that both cyanotoxins would induce toxic effects, although the influence of their combination on their toxicity is still unknown. In the present study, the potential mutagenicity of CYN was studied using the Bacterial Reverse Mutation Test (OECD 471). In addition, the cytotoxic effects of the combination of different concentrations of CYN and MC-LR were studied in the hepatic cell line HepG2 through the MTS reduction assay. This combination was also evaluated using the CalcuSyn program in order to establish an additive or synergistic effect of the toxins. Preliminary results did not show a mutagenic response produced by CYN. With respect to the cytotoxicity, the mixture appeared to induce more damage than the cyanotoxins individually. Further works to clarify the mutagenic potential of CYN and how simultaneous exposure to cyanotoxins affects their toxicity are needed. Acknowledgements: The authors want to thank the Spanish Ministerio de Economía y Competitividad (AGL2015-64558-R) for the financial support. Moreover, the authors want to thank the Biology Service of the CITIUS of the University of Sevilla for their technical support. http://dx.doi.org/10.1016/j.toxlet.2016.06.1615 P09-014 Novel graphical tool to synthetise evidence from animal studies reporting on toxicological endpoints C. Croera 1,∗ , R. Pirow 2 , U. Gundert Remy 3 , T. Husøy 2 , C. Smeraldi 1 , A. Castoldi 1 1
European Food Safety Authority, Food Ingredients and Packaging Unit (FIP), Parma, Italy 2 Working Group on Bisphenol A Toxicology, European Food Safety Authority (EFSA), Parma, Italy 3 Working Group on Isoflavones, European Food Safety Authority (EFSA), Parma, Italy In 2015 the CEF and ANS EFSA Scientific Panels published the opinions on: (1) the health risks from bisphenol A (BPA) in foodstuffs, and on (2) the potential harmful effects of isoflavones in food supplements for peri- and post-menopausal women. For both assessments, a large amount of data was gathered from literature. The data were too heterogeneous for a formal meta-analysis. Hence, a novel graphical approach was developed to synthesise the data on specific endpoints and explore the dose–response relationship. The graphical figures were generated in the statistical computing envi-
S163
ronment R using an adapted code based on the R Package ‘lattice’. In BPA assessment, “Proliferative changes in mammary gland” were identified as toxicity endpoint. The related animal studies were grouped according to the histopathological changes observed. For isoflavones, “Increase of uterine weight” in ovariectomised (OVX) animals was identified as an endpoint. The studies were grouped according to the type of isoflavones administered and sorted for their duration. For BPA, the results were plotted on a common oral human equivalent dose scale. The graph included scores for study reliability and relevance, strengths and weaknesses, exposure and time of assessment. For isoflavones, statistical significant increase in uterine weight in treated animals versus OVX-control was indicated by the size of the dot, being proportionate to the relative effect compared to OVX-control. This graphical tool can be an effective way for synthesising evidence from a large number of animal studies reporting on the same endpoint. http://dx.doi.org/10.1016/j.toxlet.2016.06.1616 P09-015 Assessment of mercury intake based on intervention research in Polish subpopulation R. Kuras ∗ , B. Janasik, M. Stanislawska, W. Wasowicz Department of Biological and Environmental Monitoring, Nofer Institute of Occupational Medicine, Lodz, Poland Fish and marine mammal consumptions are an important pathway for human exposure to mercury (Hg), and ingestion of fish contaminated with methylmercury can lead to adverse health outcomes. In this study, we analyzed mercury content in hair and blood samples collected from 67 volunteers during June to July 2015. An intervention study was based on 10-day fish consumption frequency. Hg concentrations in hair and blood samples derived from men were determined using TDA-AAS technique. Mean concentration of Hg in whole blood of volunteers in the first day of fish consumption was found to be: 0.62 ± 0.41 g/l (NS); after one week: 0.90 ± 0.46 g/l (p < 0.001) and in the end of the study 1.28 ± 0.49 g/l (p < 0.001) and one month after the end of the study (“wash–out” period) 0.78 ± 0.60 g/l (p < 0.001). Mean concentration of Hg in hair of volunteers in the first day of fish consumption was found to be: 0.24 ± 0.16 g/l (NS) and wash–out period: 0.29 ± 0.15 g/l (p < 0.05). The paper estimate weekly Hg intake from fish based on intervention study. The projected intake values of Hg through human consumption were calculated and were compared with the PTWI value established by JECFA. Acknowledgement: Financial support by Grant: 2013/11/B/NZ7/04934. http://dx.doi.org/10.1016/j.toxlet.2016.06.1617 P09-016 Metals that are important for food safety control of bread product Y. Feyzi 1 , A. Malekirad 1,∗ , M. Fazilati 1 , H. Salavati 1 , S. Habibollahi 1 , M. Rezaei 2 1
Department of Biology, Payame Noor University, Tehran, Iran Department of Food Safety and Hygiene, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2
Introduction: Breads are among the most important elements of our daily diet. Knowing about the heavy and essential elements
P09-013 In vitro study of mutagenicity of cylindrospermopsin and cytotoxic effects of the combination of microcystin and cylindrospermopsin A. Jos ∗ , D. Gutiérrez Praena, S. Maisanaba, M. Llana Ruíz Cabello, A. Cameán Area of Toxicology, Faculty of Pharmacy, University of Sevilla. C/Profesor García González 2, 41012 Sevilla, Spain Cylindrospermopsin (CYN) and microcystin-LR (MC-LR) are two cyanotoxins produced by several species of cyanobacteria and they are well distributed worldwide. It is known these toxins, separately, induce harmful effects in human, animals and plants, included those destined for human consumption. However, some of their toxic effects are still scarcely investigated, such as the mutagenicity induced by CYN. Moreover, CYN and MC-LR can appear in water reservoirs at the same time, so it could be assumed that both cyanotoxins would induce toxic effects, although the influence of their combination on their toxicity is still unknown. In the present study, the potential mutagenicity of CYN was studied using the Bacterial Reverse Mutation Test (OECD 471). In addition, the cytotoxic effects of the combination of different concentrations of CYN and MC-LR were studied in the hepatic cell line HepG2 through the MTS reduction assay. This combination was also evaluated using the CalcuSyn program in order to establish an additive or synergistic effect of the toxins. Preliminary results did not show a mutagenic response produced by CYN. With respect to the cytotoxicity, the mixture appeared to induce more damage than the cyanotoxins individually. Further works to clarify the mutagenic potential of CYN and how simultaneous exposure to cyanotoxins affects their toxicity are needed. Acknowledgements: The authors want to thank the Spanish Ministerio de Economía y Competitividad (AGL2015-64558-R) for the financial support. Moreover, the authors want to thank the Biology Service of the CITIUS of the University of Sevilla for their technical support. http://dx.doi.org/10.1016/j.toxlet.2016.06.1615 P09-014 Novel graphical tool to synthetise evidence from animal studies reporting on toxicological endpoints C. Croera 1,∗ , R. Pirow 2 , U. Gundert Remy 3 , T. Husøy 2 , C. Smeraldi 1 , A. Castoldi 1 1
European Food Safety Authority, Food Ingredients and Packaging Unit (FIP), Parma, Italy 2 Working Group on Bisphenol A Toxicology, European Food Safety Authority (EFSA), Parma, Italy 3 Working Group on Isoflavones, European Food Safety Authority (EFSA), Parma, Italy In 2015 the CEF and ANS EFSA Scientific Panels published the opinions on: (1) the health risks from bisphenol A (BPA) in foodstuffs, and on (2) the potential harmful effects of isoflavones in food supplements for peri- and post-menopausal women. For both assessments, a large amount of data was gathered from literature. The data were too heterogeneous for a formal meta-analysis. Hence, a novel graphical approach was developed to synthesise the data on specific endpoints and explore the dose–response relationship. The graphical figures were generated in the statistical computing envi-
S163
ronment R using an adapted code based on the R Package ‘lattice’. In BPA assessment, “Proliferative changes in mammary gland” were identified as toxicity endpoint. The related animal studies were grouped according to the histopathological changes observed. For isoflavones, “Increase of uterine weight” in ovariectomised (OVX) animals was identified as an endpoint. The studies were grouped according to the type of isoflavones administered and sorted for their duration. For BPA, the results were plotted on a common oral human equivalent dose scale. The graph included scores for study reliability and relevance, strengths and weaknesses, exposure and time of assessment. For isoflavones, statistical significant increase in uterine weight in treated animals versus OVX-control was indicated by the size of the dot, being proportionate to the relative effect compared to OVX-control. This graphical tool can be an effective way for synthesising evidence from a large number of animal studies reporting on the same endpoint. http://dx.doi.org/10.1016/j.toxlet.2016.06.1616 P09-015 Assessment of mercury intake based on intervention research in Polish subpopulation R. Kuras ∗ , B. Janasik, M. Stanislawska, W. Wasowicz Department of Biological and Environmental Monitoring, Nofer Institute of Occupational Medicine, Lodz, Poland Fish and marine mammal consumptions are an important pathway for human exposure to mercury (Hg), and ingestion of fish contaminated with methylmercury can lead to adverse health outcomes. In this study, we analyzed mercury content in hair and blood samples collected from 67 volunteers during June to July 2015. An intervention study was based on 10-day fish consumption frequency. Hg concentrations in hair and blood samples derived from men were determined using TDA-AAS technique. Mean concentration of Hg in whole blood of volunteers in the first day of fish consumption was found to be: 0.62 ± 0.41 g/l (NS); after one week: 0.90 ± 0.46 g/l (p < 0.001) and in the end of the study 1.28 ± 0.49 g/l (p < 0.001) and one month after the end of the study (“wash–out” period) 0.78 ± 0.60 g/l (p < 0.001). Mean concentration of Hg in hair of volunteers in the first day of fish consumption was found to be: 0.24 ± 0.16 g/l (NS) and wash–out period: 0.29 ± 0.15 g/l (p < 0.05). The paper estimate weekly Hg intake from fish based on intervention study. The projected intake values of Hg through human consumption were calculated and were compared with the PTWI value established by JECFA. Acknowledgement: Financial support by Grant: 2013/11/B/NZ7/04934. http://dx.doi.org/10.1016/j.toxlet.2016.06.1617 P09-016 Metals that are important for food safety control of bread product Y. Feyzi 1 , A. Malekirad 1,∗ , M. Fazilati 1 , H. Salavati 1 , S. Habibollahi 1 , M. Rezaei 2 1
Department of Biology, Payame Noor University, Tehran, Iran Department of Food Safety and Hygiene, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2
Introduction: Breads are among the most important elements of our daily diet. Knowing about the heavy and essential elements