Analysis of the sensitivity of in vitro bioassays for androgenic, progestagenic, glucocorticoid, thyroid and estrogenic activity: Suitability for drinking and environmental waters.

doi:10.1016/J.ENVINT.2016.12.014

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Analysis of the sensitivity of in vitro bioassays for androgenic, progestagenic, glucocorticoid, thyroid and estrogenic activity: Suitability for drinking and environmental waters.

Journal Article•DOI•

Analysis of the sensitivity of in vitro bioassays for androgenic, progestagenic, glucocorticoid, thyroid and estrogenic activity: Suitability for drinking and environmental waters.

Frederic D.L. Leusch¹, Peta A. Neale¹, Armelle Hebert, Marco Scheurer, Merijn Schriks - Show less +1 more•Institutions (1)

Griffith University¹

01 Feb 2017-Environment International (Environ Int)-Vol. 99, pp 120-130

TL;DR: Assessing whether current in vitro bioassays are suitable to detect endocrine activity in a range of water types can help provide guidance on in vitroBioassay selection and required sample enrichment for optimised detection of endocrineactivity in environmental waters.

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About: This article is published in Environment International.The article was published on 2017-02-01. It has received 71 citations till now. The article focuses on the topics: Bioassay.

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Journal Article•DOI•

Effect-based trigger values for in vitro and in vivo bioassays performed on surface water extracts supporting the environmental quality standards (EQS) of the European Water Framework Directive.

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Beate I. Escher, Selim Aїt-Aїssa, Peter A. Behnisch, Werner Brack¹, Werner Brack², François Brion, Abraham Brouwer, Sebastian Buchinger, Sarah E. Crawford², David Du Pasquier, Timo Hamers³, Karina Hettwer, Klára Hilscherová⁴, Henner Hollert², Robert Kase⁵, Cornelia Kienle⁵, Andrew J. Tindall, Jochen Tuerk, Ron van der Oost, Etiënne L.M. Vermeirssen⁵, Peta A. Neale⁶, Peta A. Neale⁷ - Show less +18 more•Institutions (7)

Helmholtz Centre for Environmental Research - UFZ¹, RWTH Aachen University², VU University Amsterdam³, Masaryk University⁴, Swiss Federal Institute of Aquatic Science and Technology⁵, University of Queensland⁶, Griffith University⁷

01 Jul 2018-Science of The Total Environment

TL;DR: A method that reads across from existing EQS and includes additional mixture considerations with the goal that the derived effect-based trigger values (EBT) indicate acceptable risk for complex mixtures as they occur in surface water.

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167 citations

Journal Article•DOI•

Analysis of endocrine activity in drinking water, surface water and treated wastewater from six countries

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Frederic D.L. Leusch¹, Peta A. Neale¹, Charlotte Arnal, N. H. Aneck-Hahn², Patrick Balaguer³, Auguste Bruchet⁴, Beate I. Escher⁵, Beate I. Escher¹, Beate I. Escher⁶, M. Esperanza⁴, Marina Grimaldi³, Gaëla Leroy, Marco Scheurer, Rita Schlichting⁶, Merijn Schriks, Armelle Hebert - Show less +12 more•Institutions (6)

Griffith University¹, University of Pretoria², University of Montpellier³, Suez Environnement⁴, University of Tübingen⁵, Helmholtz Centre for Environmental Research - UFZ⁶

01 Aug 2018-Water Research

TL;DR: Some of the wastewater and surface water samples were found to exceed the effect-based trigger values for estrogenic and glucocorticoid activity, suggesting these environmental waters may pose a potential risk to ecosystem health, while the lack of bioassay activity and low detected chemical concentrations in the drinking water samples do not suggest a risk to human endocrine health.

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87 citations

Cites background from "Analysis of the sensitivity of in v..."

...Estrogenic activity in wastewater effluent will vary depending on the treatment processes and wastewater type (Maletz et al., 2013; Valitalo et al., 2016), with surface waters downstream of wastewater discharges often having elevated estrogenicity (Schiliro et al., 2009; Mnif et al., 2012)....
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...Estrogenic activity in wastewater effluent will vary depending on the treatment processes and wastewater type (Maletz et al., 2013; Valitalo et al., 2016), with surface waters downstream of wastewater discharges often having elevated estrogenicity (Schiliro et al....
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Journal Article•DOI•

Screening and risk management solutions for steroidal estrogens in surface and wastewater

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Robert Kase¹, Barbora Javurkova, Eszter Simon¹, Kees Swart, Sebastian Buchinger, Sarah Könemann², Sarah Könemann¹, Beate I. Escher³, Mario Carere, Valeria Dulio, Selim Ait-Aissa, Henner Hollert², Sara Valsecchi⁴, Stefano Polesello⁴, Peter A. Behnisch, Carolina Di Paolo², Daniel Olbrich¹, Eliška Sychrová, Michael Gundlach², Rita Schlichting³, Lomig Leborgne, Manfred Clara, Christoph Scheffknecht, Yves Marneffe, Carole Chalon, Petr Tusil, Premysl Soldan, Brigitte von Danwitz, Julia Schwaiger, Antonio Moran Palao⁵, Francesca Bersani, Olivier Perceval, Cornelia Kienle¹, Etiënne L.M. Vermeirssen¹, Klára Hilscherová, Georg Reifferscheid, Inge Werner¹ - Show less +33 more•Institutions (5)

Swiss Federal Institute of Aquatic Science and Technology¹, RWTH Aachen University², Helmholtz Centre for Environmental Research - UFZ³, National Research Council⁴, Spanish National Research Council⁵

01 May 2018-Trends in Analytical Chemistry

TL;DR: Konemann et al. as discussed by the authors compared bioanalytical and chemical analytical results with regard to their application for aquatic status assessment, and concluded that water quality assessment can progress from a purely analytical approach to effect-based monitoring, from single substance to known and unknown mixture assessment and from in-vitro screening to population-relevant risk assessment.

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Abstract: Background The European Commission Implementing Decision EU 2015/495 included three steroidal estrogens, namely 17α-ethinyl estradiol, 17β-estradiol, and estrone, in the so-called “watch list” of the EU Water Framework Directive (WFD). The monitoring of these compounds is difficult because the detection limits of the majority of the available analytical methods cannot achieve the very low target concentrations required to meet proposed environmental quality criteria. In 2014, a combined Science-Policy Interface/Chemical Monitoring of Emerging Pollutants project was launched to meet this monitoring challenge. The project involved 24 research organizations and environmental agencies from 12 different countries. Methods Sixteen surface water (SW) and 17 wastewater (WW) samples were collected across Europe and analysed using five in vitro effect-based and three chemical analytical methods. A general description of the project and data evaluation is provided by Konemann and colleagues in the companion publication 2018. In our study, we compared bioanalytical and chemical analytical results with regard to their application for aquatic status assessment. Therefore we considered the potential to predict population-relevant risks for aquatic organisms and the specificity and sensitivity of the various methods used in both approaches. Finally, we tested and discussed the applicability and relevance of previously suggested effect-based trigger values (EBT). Results and discussion Results of the risk assessment based on chemical analytical data correlated highly with estrogenic activities (expressed as 17β-estradiol equivalents (EEQ) determined using effect-based methods), demonstrating the ability of the bioassays to predict the mixture risk caused by steroidal estrogens. For about 15% of SW and 40% of WW samples detection limits of chemical-analytical methods were too high to allow a status assessment, while detection limits of all effect-based methods were below proposed EBT. This demonstrates that effect-based methods are suitable for status assessment of surface waters. The in vitro effect-based methods were quite specific for steroidal estrogens and highly sensitive, meaning they have a low probability to detect false positive or negative results. After testing of three alternative EBT proposals, we concluded to use preliminary 400 pg/L EEQ as screening EBT corresponding to the AA-EQS of E2. Further test specific refinements in the application of this value are not excluded. Conclusions We conclude that water quality assessment can progress from a purely analytical approach to effect-based monitoring, from single substance to known and unknown mixture assessment and from in vitro screening to population-relevant risk assessment. Despite a few limitations, effect-based in vitro methods are recommendable for monitoring steroidal estrogens under the WFD because they, a) are able to sensitively quantify the activity of steroidal estrogens in all surface and wastewater samples, b) are able to detect the combined effect of estrogen mixtures including unknown chemicals with estrogen receptor activating properties, c) allow an ecotoxicological status assessment using EBT to calculate risk quotients. This approach is similar to the risk ratio used in regulatory environmental risk assessments, but allows for an integrated mixture assessment. Outlook The results of this study support the introduction of a holistic approach for the regulation of chemicals in the aquatic environment under the EU WFD, as proposed recently by EU Water Directors. An ecotoxicological status assessment for one of the most relevant modes of action of endocrine disruption will allow authorities responsible for water quality assessment to focus available monitoring resources and to improve the ecological status of EU waterbodies.

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66 citations

Journal Article•

Critical parameters of the MCF-7 cell proliferation bioassay (E-Screen)

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T. H. Rasmussen, J. B. Nielsen

01 Jan 2001-Toxicology

TL;DR: Using this optimized test regimen, the responsiveness of treated MCF-7 BUS cells was consistently increased up to 11-fold over hormone-free controls and the specificity was characterized by examining the effects of oestradiol-17β, the anti-oestrogen ICI 182,780, and dieldrin, a recognized xeno-Oestrogen.

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64 citations

Journal Article•DOI•

Effect-based assessment of toxicity removal during wastewater treatment

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Pia Välitalo¹, Pia Välitalo², Riccardo Massei³, Ilse Heiskanen², Peter A. Behnisch, Werner Brack³, Andrew J. Tindall, David Du Pasquier, Eberhard Küster³, Anna Mikola¹, Tobias Schulze³, Markus Sillanpää² - Show less +8 more•Institutions (3)

Aalto University¹, Finnish Environment Institute², Helmholtz Centre for Environmental Research - UFZ³

01 Dec 2017-Water Research

TL;DR: It is demonstrated that a biotest battery comprising of multiple endpoints can serve as a powerful tool when assessing water quality or water treatment efficiency in a holistic manner and can be used to complement traditional methods of monitoring in the future by assessing sum-parameter based effects.

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64 citations

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Journal Article•DOI•

State of the Science of Endocrine Disrupting Chemicals - 2012

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Åke Bergman¹, Jerrold J. Heindel², Susan Jobling³, Karen A. Kidd⁴, R. Thomas Zoeller⁵ - Show less +1 more•Institutions (5)

Stockholm University¹, National Institutes of Health², Brunel University London³, University of New Brunswick⁴, University of Massachusetts Amherst⁵

17 Jun 2012-Toxicology Letters

TL;DR: The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of UNEP or WHO concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries.

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1,192 citations

"Analysis of the sensitivity of in v..." refers background in this paper

...In addition to natural and synthetic hormones, awide range of environmental chemicals, including industrial compounds, pesticides and UV filters, have been identified as known or suspected endocrine disrupting chemicals (Bergman et al., 2012)....
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Journal Article•DOI•

Several environmental oestrogens are also anti-androgens

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P Sohoni¹, John P. Sumpter•Institutions (1)

Brunel University London¹

01 Sep 1998-Journal of Endocrinology

TL;DR: It is shown that many of the so-called 'environmental oestrogens' also possess anti-androgenic activity, demonstrating that hormone-mimicking chemicals can have multiple hormonal activities, which may make it difficult to interpret their mechanisms of action in vivo.

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Abstract: There is presently considerable interest in endocrine disruption which is a new area of endocrinology concerned with chemicals that mimic hormones, in particular sex steroids. It has been hypothesised that exposure to such chemicals may be responsible for adverse effects in both humans and wildlife. Until now, chemicals that mimic oestrogens (so-called xenoestrogens) have been the main focus of endocrine disruption research. However, recent evidence suggests that many abnormalities in the male reproductive system may be mediated via the androgen receptor. By blocking androgen action, exposure to an anti-androgen may cause changes similar to those associated with oestrogen exposure. We have used in vitro yeast-based assays to detect oestrogenic, anti-oestrogenic, androgenic and anti-androgenic activities in a variety of chemicals of current interest. We show that many of the so-called 'environmental oestrogens' also possess anti-androgenic activity. The previously reported anti-androgenic activities of vinclozolin and p,p'-1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (DDE) were confirmed. We also found that o,p'-1,1,1,-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), bisphenol A and butyl benzyl phthalate were anti-androgenic. However, not all xenoestrogens are also anti-androgenic, because nonylphenol was found to be a weak androgen agonist. Our results demonstrate that hormone-mimicking chemicals can have multiple hormonal activities, which may make it difficult to interpret their mechanisms of action in vivo. Although not a specific objective of this study, our results also demonstrate that yeast-based assays are powerful tools with which to investigate both agonist and antagonistic hormonal activities of chemicals.

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842 citations

"Analysis of the sensitivity of in v..." refers background or methods in this paper

...Receptor binding assays, including the classical (Leusch et al., 2006) and recombinant androgen receptor binding assay (Freyberger and Ahr, 2004), and yeast reporter gene assays, including the classical yeast androgen screen (YAS) (Gaido et al., 1997; Sohoni and Sumpter, 1998) and two-hybrid assay (Li et al., 2008a) had similar MDLs in agonist mode, which ranged from 8.3–26 ng/L (Table 1)....
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...…receptor binding assay (Leusch et al., 2006) and the yeast reporter gene assays, yeast estrogen screen (YES) originally from John Sumpter's lab (Sohoni and Sumpter, 1998) and from Kevin Gaido's lab (Gaido et al., 1997), respectively, had similar MDLs, ranging from 1.2–1.8 ng/L. Re-analysis of…...
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...0 amiodarone hydrochloride equivalents; DF, dilution factor; DHT, dihydrotestosterone; DHTEQ, dihydrotestosterone equivalents; ; Dexa, dexamethasone; DexaEQ, dexamethasone equivalents; E2, 17β-estradiol; EEQ, 17β-estradiol equivalents; EC, effect EQ, flutamide equivalents; FulvestrantEQ, fulvestrant equivalents; Levo, levonorgestrel; LevoEQ, levonorgestrel equivalents; valents; OHFLU, hydroxyflutamide; OHFLUEQ, hydroxyflutamide equivalents; OHTMX, hydroxytamoxifen; ORG2058, 16αrogesterone; P4EQ, progesterone equivalents; PMG, promegestone; PMGEQ, promegestone equivalents; REP, relative effect tion; T3, triiodothyronine; T3EQ, triiodothyronine equivalents; TMX, tamoxifen; TMXEQ, tamoxifen equivalents; YAS, yeast )....
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..., 2006) and the yeast reporter gene assays, yeast estrogen screen (YES) originally from John Sumpter's lab (Sohoni and Sumpter, 1998) and from Kevin Gaido's lab (Gaido et al....
[...]
...…equivalents, the higher tolerance of yeast-based assays to solvents (up to 1%, compared to 0.1% for most mammalian assays) meant that the MDL for the most sensitive yeast assay (Sohoni and Sumpter, 1998) was comparable (although in the upper range) to the mammalian reporter gene assays....
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Journal Article•DOI•

Quantitation of Transcription and Clonal Selection of Single Living Cells with β-Lactamase as Reporter

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Gregor Zlokarnik, Paul A. Negulescu¹, Thomas E. Knapp¹, Lora Mere¹, Neal Burres¹, Luxin Feng¹, Michael A. Whitney¹, Klaus Roemer¹, Roger Y. Tsien¹ - Show less +5 more•Institutions (1)

University of California, San Diego¹

02 Jan 1998-Science

TL;DR: The robust change in emission ratio reveals quantitative heterogeneity in real-time gene expression, enables clonal selection by flow cytometry, and forms a basis for high-throughput screening of pharmaceutical candidate drugs in living mammalian cells.

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Abstract: Gene expression was visualized in single living mammalian cells with β-lactamase as a reporter that hydrolyzes a substrate loaded intracellularly as a membrane-permeant ester. Each enzyme molecule changed the fluorescence of many substrate molecules from green to blue by disrupting resonance energy transfer. This wavelength shift was detectable by eye or color film in individual cells containing less than 100 β-lactamase molecules. The robust change in emission ratio reveals quantitative heterogeneity in real-time gene expression, enables clonal selection by flow cytometry, and forms a basis for high-throughput screening of pharmaceutical candidate drugs in living mammalian cells.

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735 citations

"Analysis of the sensitivity of in v..." refers methods in this paper

...As mentioned above, the GeneBLAzer assays produce enzyme beta-lactamase in the presence of agonists and beta-lactamase cleaves the fluorescent substrate to increase blue fluorescence (Zlokarnik et al., 1998)....
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Journal Article•DOI•

Evaluation of Chemicals with Endocrine Modulating Activity in a Yeast-Based Steroid Hormone Receptor Gene Transcription Assay

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Kevin W. Gaido¹, Linda S. Leonard¹, Stephanie Lovell¹, Janet C. Gould¹, Dariouch Babaı̈¹, Christopher J. Portier¹, Donald P. McDonnell² - Show less +3 more•Institutions (2)

Research Triangle Park¹, Duke University²

01 Mar 1997-Toxicology and Applied Pharmacology

TL;DR: The utility of using yeast-based receptor assays for detecting chemical interaction with steroid receptors is demonstrated and these assays should serve as a useful component of an in vitro-in vivo strategy to assess the effects of chemicals on endocrine function.

...read moreread less

662 citations

"Analysis of the sensitivity of in v..." refers background or methods in this paper

...Receptor binding assays, including the classical (Leusch et al., 2006) and recombinant androgen receptor binding assay (Freyberger and Ahr, 2004), and yeast reporter gene assays, including the classical yeast androgen screen (YAS) (Gaido et al., 1997; Sohoni and Sumpter, 1998) and two-hybrid assay (Li et al., 2008a) had similar MDLs in agonist mode, which ranged from 8.3–26 ng/L (Table 1)....
[...]
...0 amiodarone hydrochloride equivalents; DF, dilution factor; DHT, dihydrotestosterone; DHTEQ, dihydrotestosterone equivalents; ; Dexa, dexamethasone; DexaEQ, dexamethasone equivalents; E2, 17β-estradiol; EEQ, 17β-estradiol equivalents; EC, effect EQ, flutamide equivalents; FulvestrantEQ, fulvestrant equivalents; Levo, levonorgestrel; LevoEQ, levonorgestrel equivalents; valents; OHFLU, hydroxyflutamide; OHFLUEQ, hydroxyflutamide equivalents; OHTMX, hydroxytamoxifen; ORG2058, 16αrogesterone; P4EQ, progesterone equivalents; PMG, promegestone; PMGEQ, promegestone equivalents; REP, relative effect tion; T3, triiodothyronine; T3EQ, triiodothyronine equivalents; TMX, tamoxifen; TMXEQ, tamoxifen equivalents; YAS, yeast )....
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...Environment International xxx (2016) xxx–xxx EI-03541; No of Pages 11 Contents lists available at ScienceDirect Environment International j ourna l homepage: www.e lsev ie r .com/ locate /env int Review article Analysis of the sensitivity of in vitro bioassays for androgenic, progestagenic, glucocorticoid, thyroid and estrogenic activity: Suitability for drinking and environmental waters Frederic D.L. Leusch a,⁎, Peta A. Neale a, Armelle Hebert b, Marco Scheurer c, Merijn C.M. Schriks d a Australian Rivers Institute, Griffith School of Environment, Griffith University, Gold Coast, Qld, 4222, Australia b Veolia Research & Innovation, Paris, France c DVGW – TechnologiezentrumWasser, Karlsruhe, Germany d KWR Watercycle Research Institute, Nieuwegein, The Netherlands Abbreviations:AH, amiodarone hydrochloride; AHEQ, DOC, dissolved organic carbon; DPH, diphenylhydantoin concentration; EF, enrichment factor; FLU, flutamide; FLU MDL, method detection limit; MifEQ, mifepristone equi ethyl-21-hydroxy-19-nor-pregn-4-ene-3,20-dione; P4, p potency; RU5020, promegestone; SPE, solid phase extrac androgen screen; YES, yeast estrogen screen....
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..., 1997) as these assays cannot distinguish between agonists and antagonists, with the pure agonist ICI 164,384 producing an estrogenic response in the YES assay (Gaido et al., 1997)....
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...…yeast reporter gene assays, yeast estrogen screen (YES) originally from John Sumpter's lab (Sohoni and Sumpter, 1998) and from Kevin Gaido's lab (Gaido et al., 1997), respectively, had similar MDLs, ranging from 1.2–1.8 ng/L. Re-analysis of E2 concentration-effect curves for ERCALUX (Sonneveld…...
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Journal Article•DOI•

An integrated assessment of estrogenic contamination and biological effects in the aquatic environment of The Netherlands

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A. Dick Vethaak, Joost Lahr¹, S. Marca Schrap, A.C. Belfroid², G.B.J. Rijs, A.A.M. Gerritsen, Jacob de Boer, Astrid S. Bulder, Guy C. M. Grinwis³, Raoul V. Kuiper³, Juliette Legler¹, Tinka A.J. Murk¹, Willie J.G.M. Peijnenburg, Henk J. M. Verhaar, Pim de Voogt² - Show less +11 more•Institutions (3)

Wageningen University and Research Centre¹, University of Amsterdam², Utrecht University³

01 Apr 2005-Chemosphere

TL;DR: It is concluded that hormones (especially 17 alpha-ethynylestradiol) and possibly also nonylphenol(ethoxylate)s are primarily responsible for these effects of feminizing effects in male fish.

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529 citations