Bioanalytical Tests for Assessing Cytotoxicity and Estrogenicity Effects of Treated Wastewater on Mammalian Cell Lines

A range of toxicity bioassays which account for the biological effects of the pollutants has been developed to assess the potential toxicity risk of chemical mixtures against living cells. The current study aimed to determine the cytotoxic and estrogenic effects of treated wastewater effluents discharging into a main River in Tunisia. Modified E-screen assay was used to evaluate the estrogenic activity of water samples on MCF-7 breast cancer cells while MTT assay was performed to assess cytotoxicity on human intestinal Caco-2 cells. According to preliminary chemical analysis, high organic pollution was detected for some samples of treated wastewater since high levels of chemical oxygen demand exceeded the Tunisian guidelines. On one hand, significant cytotoxicity on Caco-2 human intestinal epithelial cells was registered in a dose dependant manner for 0.1%, 1%, 5% and 10% of water samples concentrations. In fact, MTT results showed significant cell viability decrease reaching 46% for the highest sample concentration. On the other hand, Modified E-screen assay revealed the presence of estrogenic compounds in the effluent samples. Estrogenic potencies induced a significant proliferation level of MCF-7 cells in a dose dependent manner. Thus, water samples may contain diversity of organic contaminants considered as potentially hazardous complex mixtures and representing a potential environmental risk for surface water. The implemented assays pointed out the insufficiency of the applied treatment technology to completely eliminate toxic micropollutants from treated effluent which may threaten aquatic life and human health as well.