Abstract
Important toxicological achievements have been made during the last decades using reptiles. We focus our investigation on gonadal reproductive health of the soil biosentinel Podarcis sicula which is very sensitive to endocrine-disrupting chemicals. The aim of this study is to quantitatively detect, by sensitive microassays, reactive oxygen species and the glutathione antioxidants in the testis and investigate if they are differentially expressed before and after remediation of a site of the “Land of Fires” (Campania, Italy) subject to illicit dumping of unknown material. The oxidative stress level was evaluated by electron spin resonance spectroscopy applying a spin-trapping procedure able to detect products of lipid peroxidation, DNA damage and repair by relative mobility shift, and poly(ADP-ribose) polymerase enzymatic activity, respectively, the expression of glutathione peroxidase 4 transcript by real-time quantitative PCR analysis, the antioxidant glutathione S-transferase, a well-assessed pollution index, by enzymatic assay and the total soluble antioxidant capacity. Experimental evidences from the different techniques qualitatively agree, thus confirming the robustness of the combined experimental approach. Collected data, compared to those from a reference unpolluted site constitute evidence that the reproductive health of this lizard is impacted by pollution exposure. Remediation caused significant reduction of reactive oxygen species and downregulation of glutathione peroxidase 4 mRNAs in correspondence of reduced levels of glutathione S-transferase, increase of antioxidant capacity, and repair of DNA integrity. Taken together, our results indicate directions to define new screening approaches in remediation assessment.
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Acknowledgements
This paper gives part of the results of the action C2 of the LIFE11/ENV/IT/275 ECOREMED: Implementation of eco-compatible protocols for agricultural soil remediation in Litorale Domizio-Agro Aversano NIPS (Coordinator Prof. M. Fagnano). The authors thank the Interdepartmental Research Center for Environment (I.R.C.Env.), Federico II University, Naples, Italy, the coworkers Anna Rita Bianchi, Martina Caressa, Francesca Castaldo, Rosa D’Angelo, and Giulia Pastore for their logistics help and technical support and the colleague Dr. Anna De Maio for the helpful scientific collaboration.
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Supported by LIFE11 ENV/IT/275 (CUP E69E12000590006).
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Fig. A1
EPR spectrum of PBN adduct of radicals deriving from Podarcis sicula testis (GIF 98 kb)
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Guerriero, G., D’Errico, G., Di Giaimo, R. et al. Reactive oxygen species and glutathione antioxidants in the testis of the soil biosentinel Podarcis sicula (Rafinesque 1810). Environ Sci Pollut Res 25, 18286–18296 (2018). https://doi.org/10.1007/s11356-017-0098-8
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DOI: https://doi.org/10.1007/s11356-017-0098-8