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Toxic Effects of Wastewaters Collected at Upstream and Downstream Sites of a Purification Station in Cultures of Rainbow Trout Hepatocytes

  • C. Risso-de Faverney
  • A. Devaux
  • M. Lafaurie
  • J. P. Girard
  • R. Rahmani

Abstract

The toxic effects of wastewater samples, collected in December 1998, from upstream (U) and downstream (D) sites of the purification station of the town of Nice (South-East France on the coast of the Mediterranean Sea) were assessed undiluted and at various dilutions (75%, 50%, and 25% of collected water sample), on trout hepatocyte cultures treated for 48 or 72 h. Chemical contamination (PCBs, PAHs, Cd, Cu, Pb, and Zn) was also evaluated by chemical analysis. The water samples from the upstream site were more cytotoxic than those from the downstream site. The induction of CYP1A enzyme and metallothioneins (MTs) were selected as specific indicators of exposure to organic contaminants and metals, respectively. CYP1A-related EROD activity as well as protein expression were found to be greatly induced after 72 h exposure of the hepatocytes to the undiluted water samples (U100% and D100%), but CYP1A1 mRNA was significantly overexpressed only by samples from the upstream site. Maximal MT levels were reached after 48 h of treatment with the least concentrated water samples (U25% and D25%). Glutathione S-transferase (GST) activities were similarly increased under the same conditions. On the other hand, there was no significant glutathione peroxidase (GPx) activity response. Induction of apoptosis was analyzed by using as markers both the fragmentation of the nuclear DNA into oligonucleosomal-length fragments recognized as a “DNA ladder” and the activation of DEVD (Asp-Glu-Val-Asp)-dependent protease considered as the central mediator of programmed cell death. Significant DNA cleavage was only detectable after 72-h exposure to the most concentrated water samples from upstream sites (U75% and U100%). DEVD-dependent protease activities were significantly increased, mainly in cells exposed to U75% and D25% for 72 h. In addition, pollution-related DNA damage assessed by using the Comet assay was approximatively 1.5 times greater than that of the control level with the undiluted water samples U100% and D100%, after 72-h and 48-h exposure, respectively. The present study shows that such a multibiomarker-based approach could provide complementary information, for aquatic pollution monitoring, about the early biochemical effects in cells exposed to complex chemical pollution and could be considered as early warning systems to aquatic pollution.

Keywords

PAHs PCBs Comet Assay D100 Early Warning System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag New York Inc. 2001

Authors and Affiliations

  • C. Risso-de Faverney
    • 1
  • A. Devaux
    • 3
  • M. Lafaurie
    • 2
  • J. P. Girard
    • 2
  • R. Rahmani
    • 1
  1. 1.Laboratoire de Pharmaco-toxicologie Cellulaire et Moléculaire, Centre de Recherche INRA 41, Bd du Cap, BP 2078, 06606 Antibes cedex, FranceFR
  2. 2.Laboratoire de Toxicologie et Physiologie Environnementale, Faculté des Sciences de Nice, Parc Valrose 06000 Nice, FranceFR
  3. 3.Laboratoire des Sciences de l'Environnement, Ecole Nationale des Travaux Publics de l'Etat, rue Maurice Audin, 69518 Vaulx en Velin, FranceFR

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