Detection of estrogenic potency in wastewater and surface water with three in vitro bioassays

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.

[1]  I. Meerts,et al.  In vitro estrogenicity of polybrominated diphenyl ethers, hydroxylated PDBEs, and polybrominated bisphenol A compounds. , 2001, Environmental health perspectives.

[2]  A. D. Vethaak,et al.  A Novel in Vivo Bioassay for (Xeno-)estrogens Using Transgenic Zebrafish , 2000 .

[3]  N. Moore,et al.  The oestrogenic potential of the phthalate esters. , 2000, Reproductive toxicology.

[4]  H Fang,et al.  The estrogen receptor relative binding affinities of 188 natural and xenochemicals: structural diversity of ligands. , 2000, Toxicological sciences : an official journal of the Society of Toxicology.

[5]  A D Vethaak,et al.  Development of a stably transfected estrogen receptor-mediated luciferase reporter gene assay in the human T47D breast cancer cell line. , 1999, Toxicological sciences : an official journal of the Society of Toxicology.

[6]  A. D. Vethaak,et al.  Analysis and occurrence of estrogenic hormones and their glucuronides in surface water and waste water in The Netherlands. , 1999, The Science of the total environment.

[7]  B. Hattum,et al.  Application of biomarkers for exposure and effect of polyhalogenated aromatic hydrocarbons in naturally exposed European otters (Lutra lutra). , 1998, Environmental toxicology and pharmacology.

[8]  Edwin J. Routledge,et al.  Identification of Estrogenic Chemicals in STW Effluent. 2. In Vivo Responses in Trout and Roach , 1998 .

[9]  J. Giesy,et al.  Reproductive impairment and induction of alkaline-labile phosphate, a biomarker of estrogen exposure, in fathead minnows (Pimephales promelas) exposed to waterborne 17β-estradiol , 1998 .

[10]  T. Zacharewski.  In Vitro Bioassays for Assessing Estrogenic Substances , 1997 .

[11]  Edwin J. Routledge,et al.  Estrogenic activity of surfactants and some of their degradation products assessed using a recombinant yeast screen , 1996 .

[12]  John P. Sumpter,et al.  Inhibition of testicular growth in rainbow trout (Oncorhynchus mykiss) exposed to estrogenic alkylphenolic chemicals , 1996 .

[13]  R. Dickson,et al.  Growth factors in breast cancer. , 1995, Endocrine reviews.

[14]  J. Schwartz,et al.  Ligand-mediated modulation of estrogen receptor conformation by estradiol analogs. , 1993, Biochemistry.

[15]  J. Legler.  Development and application of in vitro and in vivo reporter gene assays for the assessment of (xeno-)estrogenic compounds in the aquatic environment , 2001 .

[16]  Y. Magara,et al.  Endocrine Disrupters in Water. , 1999 .