• Risk assessment;
  • Wastewater;
  • Leachate;
  • Heavy metals;
  • Stress response


Heavy metals present in water environment and hazardous sites as single compounds or mixture may drastically affect human health. In the present work, we investigated the risk assessment of wastewater effluents and leachate with a focus on three heavy metals—nickel (Ni), cadmium (Cd), and lead (Pb)—and their combined effect on mammalian cells, using Chinese hamster ovary cells transfected with the heat-shock protein (HSP) 47 promoter. The heavy metal mixture model was designed based on the concentrations of metals in wastewater effluents and leachate sampled in Tunisia. Using a ternary diagram, we investigated the stress response of the interaction model. This research indicated that the single heavy metals induced the stress response on HSP(+) cells even at concentrations lower than the local and international guidelines. Differences in water quality likely influenced the metal responses such that the organic composition of the leachate increased the stress response induced by the heavy metals exclusively, whereas the effluents included organic compounds that were able to mask the heavy metal effect. The mixture characterization discovered the key role played by the high levels of Ni or combination of Cd and Pb to induce the highest stress response following 3-h incubation. Heat-shock protein 47 has proven its effectiveness for assessing the heavy metal mixture effect even at low concentrations. Furthermore, the combination of a bioassay system with a statistical model proved extremely useful for better understanding the major contributors to the stress response of the mixture. Environ. Toxicol. Chem. 2010;29:1637–1647. © 2010 SETAC