Cytotoxicity and genotoxicity of sewage treatment plant effluents in rainbow trout cells (RTG-2).

The cytotoxicity and genotoxicity of 11 organic fractions from sewage treatment plant (STP) effluents were tested using the RTG-2 rainbow trout permanent cell line. An automated in vitro micronucleus assay developed for RTG-2 cells was used to test the genotoxicity, whereas neutral red uptake, kenacid blue protein assay and ATP content were used to evaluate cytotoxicity. The induction of micronuclei (MN) and alterations in the cell cycle were analysed in these cells by flow cytometry after exposure to the organic fractions for 72 h. More than half of the organic extracts tested demonstrated a significant increase in the MN frequency, thus indicating that most of them can be considered to be genotoxic. The extracts were analysed chemically by GC/MS. Although the most frequently detected compounds in the effluents were bisphenol A (BPA), octylphenol (OP), di-2-ethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP), as well as other possible mutagens, the concentrations cannot explain the genotoxicity of the individual chemicals, thereby suggesting a mixture effect. The results obtained support the need to apply effect-based tests to monitor complex mixtures as the most accurate means of assessing the genotoxicity of environmental samples.