title: Detection of estrogenic potency in wastewater and surface water with three in vitro bioassays.
abstract: A study was performed to optimize sample preparation and application of three in vitro assays for measuring estrogenic potency in environmental extracts. The three assays applied were an estrogen receptor (ER)-binding assay and two reporter gene effect assays: a yeast estrogen screen (YES) and the ER-mediated chemically activated luciferase gene expression (ER-CALUX) assay. All assays were able to detect estrogenicity, but the amounts of material needed for the assays differed greatly between the three assays (ER-binding assay >> YES > ER-CALUX). In addition, in the ER-binding assay, both agonists and antagonists give an estrogenic response, resulting in higher estradiol equivalency (EEQ) levels than both the ER-CALUX and the YES assay for the same samples. The EEQs found in wastewater treatment plants (WTPs) with the ER-CALUX assay were in the range of 4 to 440 and 0.11 to 59 pmol/L for influent and effluent, respectively. Water extracts from four large rivers had levels ranging from 0.25 to 1.72 pmol/L. Extracts from suspended matter and sludge contained estrogenic potency of 0.26 to 2.49 and 1.6 to 41 pmol EEQ/g dry weight, respectively. In WTPs, the average reduction of estrogenic potency in effluent compared to influent was 90 to 95% in municipal WTPs and about 50% in industrial WTPs. In influent, 30% of the ER-CALUX activity could not be explained by the calculated potencies based on chemical analysis of a number of known (xeno)estrogens; in effluent the unexplained fraction was 80%. These first results of analyzing estrogenic potency in WTP water and surface water in The Netherlands indicate that further studies are warranted to investigate the actual risks for aquatic systems.
authors.affiliation: Division of Toxicology, Wageningen University, The Netherlands. tinka.murk@algemeen.tox.wau.nl
authors.forename: Albertinka J
authors.initials: AJ
authors.lastname: Murk
authors.forename: Juliette
authors.initials: J
authors.lastname: Legler
authors.forename: Marola M H
authors.initials: MM
authors.lastname: van Lipzig
authors.forename: John H N
authors.initials: JH
authors.lastname: Meerman
authors.forename: Angelique C
authors.initials: AC
authors.lastname: Belfroid
authors.forename: Albertus
authors.initials: A
authors.lastname: Spenkelink
authors.forename: Bart
authors.initials: B
authors.lastname: van der Burg
authors.forename: Gerard B J
authors.initials: GB
authors.lastname: Rijs
authors.forename: Dick
authors.initials: D
authors.lastname: Vethaak
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D001681:Biological Assay / Q000379:methods*; D004784:Environmental Monitoring / Q000379:methods*; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000032:analysis* / Q000633:toxicity; D005260:Female; D005618:Fresh Water / Q000737:chemistry; D017930:Genes, Reporter; D006801:Humans; D066298:In Vitro Techniques; D008156:Luciferases / Q000235:genetics; D009426:Netherlands; D051381:Rats; D011960:Receptors, Estrogen / Q000378:metabolism; D012680:Sensitivity and Specificity; D012722:Sewage / Q000737:chemistry; D014407:Tumor Cells, Cultured; D014599:Uterus / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000633:toxicity; D015003:Yeasts
chemical_list: D004967:Estrogens; D011960:Receptors, Estrogen; D012722:Sewage; D014874:Water Pollutants, Chemical; D004958:Estradiol; D008156:Luciferases