engleski

Cytotoxic, mutagenic and genotoxic effect of wastewater from the phosphoric gypsum depot

Growing problem in many European countries is environmental pollution with municipal and industry waste water. In this study we have investigated cytotoxic, mutagenic and genotoxic effects of wastewater from the phosphoric gypsum depot near the factory for fertilizing agents, INA Petrokemija, Kutina. For determination of cytotoxic and mutagenic effects of phosphoric gypsum, we have incubated two strains of Salmonella typhimurium TA98 and TA100 with phosphoric gypsum, in presence of S9 mix, glutathione or buffer. Time of incubation was 20 minutes. For cytotoxic effect determination of wastewater on human cells, we have treated human laryngeal carcinoma cells (HEp2) and human cervical cells (HeLa) for 72 hours with different concentrations of phosphoric gypsum. The level of lipid peroxidation in these two cell lines after 24, 48 and 72 hours of treatment with different concentrations of wastewater was measured. Alkaline comet assay was conducted after 24, 48 and 72 hours of incubation of HEp2 and HeLa cells with different concentrations of wastewater in order to determine ability of wastewater to cause DNA damage. No cytotoxic effect of phosphoric gypsum on Salmonella typhimurium strains was observed, although the number of revertants of Salmonella typhimurium TA98 were decreased in comparison to negative control pointing out that there is certain toxic effect. Wastewater did not cause point mutations in presence of S9 mix, GSH or PBS. Contrary to these results, strong cytotoxic effect was observed on both human cell lines when they were treated with different concentrations of wastewater. Lipid peroxidation was induced and increased by prolonged time of incubation, pointing out that damage was not repaired, but increased with the time of incubation. Phosphoric gypsum has caused DNA damage. We can conclude that phosphoric gypsum causes high toxicity, act as prooxidant causing free radicals formation and causes DNA damage in concentrations in which it is present in depot.

wastewater; cytotoxicity; mutagenicity; lipid peroxidation; alkaline comet assay

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