Identification of endocrine disrupting chemicals from a polluted river (Lambro) in Italy using bioassays and chemical analysis.

Recently increasing concern evolved about endocrine disrupting chemicals (EDC) in the environment. Some studies showed that these chemicals can influence the reproductive system of animals in the wildlife producing e.g. intersex phenomenon in fishes or in marine snails. In this joint project a polluted river (Lambro) was investigated as a source of EDC to the wildlife of the major Italian watercourse, the river Po. Water and sediment samples from the Lambro river were fractionated and forwarded to bioassays (MVLN cells and yeast screening assay (YES)) in order to identify (anti)estrogenic activities of fractions. All fractions have been chemically analysed qualitatively as well as quantitatively for known EDC and, in addition, an immunoassay for estradiol (E2) was used to quantify E2 amounts. A proteomics analysis of the MVLN cells was conducted from treated cells relative to the control. This combined assay using chemical and biological analyses revealed some chemicals to act estrogenic. Good coincidence was found between the assays and some fractions from the water and the sediment samples were identified as being estrogenic. The chemical analysis showed that E2, E1, E3, Bisphenol A, and teroctylphenol are the most abundant compounds in the fractions with the highest estrogenic activities. The protein pattern of MVLN cells was changed in all treated samples relative to control. We conclude that these chemicals found in the Lambro water are present in concentrations affecting the cell based assays, change the protein patterns in the MVLN cells and may therefore act as EDC in the wildlife. Further investigations will show how fishes and amphibians are affected by the Lambro water conducting morphological studies and the expression of molecular biomarkers.