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- Title
Screening complex effluents for estrogenic activity with the T47D-KBluc cell bioassay: Assay optimization and comparison with in vivo responses in fish.
- Authors
Wehmas, Leah C.; Cavallin, Jenna E.; Durhan, Elizabeth J.; Kahl, Michael D.; Martinovic, Dalma; Mayasich, Joe; Tuominen, Tim; Villeneuve, Daniel L.; Ankley, Gerald T.
- Abstract
Wastewater treatment plant (WWTP) effluents can contain estrogenic chemicals, which potentially disrupt fish reproduction and development. The current study focused on the use of an estrogen-responsive in vitro cell bioassay (T47D-KBluc), to quantify total estrogenicity of WWTP effluents. We tested a novel sample preparation method for the T47D-KBluc assay, using powdered media prepared with direct effluent. Results of the T47D-KBluc assay were compared with the induction of estrogen receptor-regulated gene transcription in male fathead minnows ( Pimephales promelas) exposed to the same effluents. Effluent samples for the paired studies were collected over the course of three months. According to the T47D-KBluc assay, the effluent estrogenicity ranged from 1.13 to 2.00 ng 17β-estradiol (E2) equivalents/L. Corresponding in vivo studies exposing male fathead minnows to 0, 10, 50, and 100% effluent dilutions demonstrated that exposure to 100% effluent significantly increased hepatic vitellogenin (VTG) and estrogen receptor α subunit transcripts relative to controls. The induction was also significant in males exposed to 250 ng E2/L or 100 ng E2/L. The in vitro and in vivo results support the conclusion that the effluent contains significant estrogenic activity, but there was a discrepancy between in vitro- and in vivo-based E2 equivalent estimates. Our results suggest that the direct effluent preparation method for the T47D-KBluc assay is a reasonable approach to estimate the estrogenicity of wastewater effluent. Environ. Toxicol. Chem. 2011;30:439-445. © 2010 SETAC
- Subjects
EFFLUENT quality; SEWAGE disposal plants; WASTEWATER treatment; FISH reproduction; FISH development; FATHEAD minnow
- Publication
Environmental Toxicology & Chemistry, 2011, Vol 30, Issue 2, p439
- ISSN
0730-7268
- Publication type
Article
- DOI
10.1002/etc.388