The influence of a surfactant, linear alkylbenzene sulfonate, on the estrogenic response to a mixture of (xeno)estrogens in vitro and in vivo

This is the post-print version of the final paper published in Aquatic Toxicology. The published article is available from the link below. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. Copyright © 2008 Elsevier B.V. All rights reserved.The effect of the presence of a surfactant on the activity of a mixture of environmental estrogens was assessed. In their natural habitat, fish are subject not only to exposure to mixtures of estrogenic compounds, as has been addressed in previous publications, but also to other confounding factors (chemical, physical and biological), which may, in theory, affect their responses to such compounds. To assess the potential for such interference, the commonly occurring surfactant, linear alkylbenzene sulfonate (LAS), was applied to the yeast estrogen screen at various concentrations, independently and together with a mixture of estrogens at constant concentrations. LAS enhanced the estrogenic activity of the mixture, an effect which became less pronounced over the course of time. This information was used to design an in vivo study to assess induction of vitellogenin in fathead minnows exposed to the same mixture of estrogens plus LAS. A similar trend was observed, that is, the response was enhanced, but the effect became less pronounced as the study progressed. However, the enhanced response in vivo occurred only at the highest concentration of LAS tested (362 μg/L), and was transient because it was no longer apparent by the end of the study. Although LAS is a significant contaminant in terms of both concentration and frequency of detection in the aquatic environment, these data do not suggest that it will have a significant impact on the response of fish to environmental estrogens

Evidence of estrogenic mixture effects on the reproductive performance of fish

The official published version can be obtained from the link below - Copyright @ 2007 American Chemical SocietyRecent research into the effects of mixtures of estrogenic chemicals has revealed the capacity for similarly acting chemicals to act in combination, according to the principles of concentration addition. This means that, collectively, they may pose a significant environmental risk, even when each component is present at a low and individually ineffective concentration. The aim of this study was to investigate the ecological significance of mixture effects at low-effect concentrations by assessing the combined effect of estrogenic chemicals on the reproductive performance of fish. Pairs of fathead minnows were exposed to five estrogenic chemicals. Endpoints analyzed included fecundity, the expression of male secondary sexual characteristics, somatic indices, and vitellogenin induction. In the first phase of the study, a concentration-response analysis was performed to investigate the relative sensitivity of these endpoints. In the second phase, mixture effects at low-effect concentrations were explored by exposing fish to each of the mixture components, individually and in combination. Data from these experiments provide evidence of mixture effects on fitness and fecundity, demonstrating the capacity for chemicals to act together to affect reproductive performance, even when each component is present belowthe threshold of detectable effects. This has important implications for hazard assessment and contributes to our understanding of mixture effects at increasing levels of biological complexity.This work was funded by the European Commission, under contract EVK1-2001-00091

First year growth in relation to prenatal exposure to endocrine disruptors - A dutch prospective cohort study

Growth in the first year of life may already be predictive of obesity later in childhood. The objective was to assess the association between prenatal exposure to various endocrine disrupting chemicals (EDCs) and child growth during the first year. Dichlorodiphenyldichloroethylene (DDE), mono(2-ethyl-5-carboxypentyl)phthalate (MECPP), mono(2-ethyl-5-hydroxyhexyl)phthalate (MEHHP), mono(2-ethyl-5-oxohexyl)phthalate (MEOHP), polychlorinated biphenyl-153, perfluorooctanesulfonic acid, and perfluorooctanoic acid were measured in cord plasma or breast milk. Data on weight, length, and head circumference (HC) until 11 months after birth was obtained from 89 mother-child pairs. Mixed models were composed for each health outcome and exposure in quartiles. For MEOHP, boys in quartile 1 had a higher BMI than higher exposed boys (p = 0.029). High DDE exposure was associated with low BMI over time in boys (0.8 kg/m2 difference at 11 m). Boys with high MECPP exposure had a greater HC (1.0 cm difference at 11 m) than other boys (p = 0.047), as did girls in the second quartile of MEHHP (p = 0.018) and DDE (p < 0.001) exposure. In conclusion, exposure to phthalates and DDE was associated with BMI as well as with HC during the first year after birth. These results should be interpreted with caution though, due to the limited sample size

Prenatal exposure to endocrine disrupting chemicals in relation to thyroid hormone levels in infants – a Dutch prospective cohort study

Background: Endocrine disrupting chemicals (EDCs) present in the environment may disrupt thyroid hormones, which in early life are essential for brain development. Observational studies regarding this topic are still limited, however as the presence of chemicals in the environment is ubiquitous, further research is warranted. The objective of the current study was to assess the association between exposure markers of various EDCs and thyroxine (T4) levels in newborns in a mother-child cohort in the Netherlands. Methods: Exposure to dichlorodiphenyldichloroethylene (DDE), three di-2-ethylhexyl phthalate (DEHP) metabolites, hexachlorobenzene (HCB), polychlorinated biphenyl (PCB)-153, perfluorooctanesulfonic acid (PFOS), and perfluorooctanoic acid (PFOA) was determined in cord plasma or breast milk, and information on T4 levels in heel prick blood spots was obtained through the neonatal screening programme in the Netherlands. Linear regression models were composed to determine associations between each of the compounds and T4, which were stratified for gender and adjusted for a priori defined covariates. Results: Mean T4 level was 86.9 nmol/L (n = 83). Girls in the highest quartile of DDE and PFOA exposure showed an increased T4 level compared to the lowest quartile with both crude and fully adjusted models (DDE > 107.50 ng/L, +24.8 nmol/L, 95% CI 0.79, 48.75; PFOA > 1200 ng/L, +38.6 nmol/L, 95% CI 13.34, 63.83). In boys a lower T4 level was seen in the second quartile of exposure for both PFOS and PFOA, however after fully adjusting the models these associations were attenuated. No effects were observed for the other compounds. Conclusion: DDE and perfluorinated alkyl acids may be associated with T4 in a sex-specific manner. These results should however be interpreted with caution, due to the relatively small study population. More research is warranted, as studies on the role of environmental contaminants in this area are still limited

Sea bass (Dicentrarchus labrax) : a model organism for assessing multi-level responses to estrogenic chemicals in marine surface waters

Resumo apresentado sob poster ao 5th International Symposium of Fish Endocrinology, CAstellon, Spain, Setember 5-9, 2004.There is growing concern that aquatic wildlife in surface waters of the European Union is exposed to natural and man-made chemicals that have the ability to mimic estrogens and lead to reproductive dysfunction. Estrogenic responses in fish are the net result of complex chains of events involving the uptake, distribution and metabolism of test agents until they interact with their target sites. Typically these aspects cannot be modelled in short-term cell-based assays, only studies with vertebrates offer the opportunity to assess potential interactions of test compounds at higher organisational levels. The most widely studied biological response in fish to environmental estrogens is the production of vitellogenin (Vtg). However, few studies have attempted to link this endpoint with effects on xenobiotic biotransformation enzymes and genotoxic responses. This work is part of a study focusing on the combination effects of mixtures of estrogenic chemicals in marine and freshwater organisms. As test organism the sea bass (Dicentrarchus labrax) was selected, a common species in European marine systems. Juveniles were exposed under a flow-through system for 14 days to the natural estrogen 17ß-estradiol and ethynylestradiol. Actual chemical concentrations in the water-column were determined by gas chromatography with ion trap detection. Effects at subcellular level were analysed using Vtg as a reference endpoint [1]. Its relevance is evaluated by further investigations on liver 7-ethoxyresorufin-O-deethylase (EROD) activity and erythrocytic nuclear abnormalities [2]. These measurements were integrated with organism level endpoints (i.e. condition factor, hepatossomatic index) to provide evidence for cause-effect of estrogenic contamination. The general suitability of the sea bass as a model organism for the screening of estrogenic chemicals in the marine environment is discussed.Comissão Europeia (CE) - ACE, EVK1-CT-2001-100

Sea bass (Dicentrarchus labrax) : a model organism for the screening of estrogenic chemicals in marine surface waters?

Society of Environmental Toxicology and Chemistry - SETAC Europe 14th Annual Meeting, Prague, Czech Republic, April 2004.There is growing concern that aquatic wildlife in surface waters of the European Union is exposed to natural and man-made chemicals that have the ability to mimic estrogens and lead to reproductive dysfunction. Estrogenic responses in fish are the net result of complex chains of events involving the uptake, distribution and metabolism of test agents until they interact with their target sites. Typically these aspects cannot be modelled in short-term cell-based assays, only studies with vertebrates offer the opportunity to assess potential interactions of test compounds at higher organisational levels. However, studies with endocrine disrupting chemicals have been performed mainly with freshwater organisms. The sensitivity of a marine fish species to different estrogenic chemicals was investigated under chronic exposure conditions. This work is part of a study focusing on the combination effects of mixtures of estrogenic chemicals in marine and freshwater organisms (ACE, EVK1-CT-2001-100). As test organism the sea bass (Dicentrarchus labrax) was selected, a common species in European marine systems. Juveniles were exposed under a flow-through system for 14 days for a set of reference chemicals (17Ã -estradiol, ethynylestradiol, nonylphenol, octylphenol, bisphenol A). Effects at subcellular level were analysed using vitellogenesis as endpoint. Its relevance is evaluated by further investigations about the individual fitness (condition factor, hepatossomatic index), as well as the liver cytochrome P450 activity. The general suitability of the sea bass as a model organism for the screening of estrogenic chemicals in the marine environment is discussed.Comissãoo Europeia (CE) - ACE, EVK1-CT-2001-100

High resolution effect-directed analysis of steroid hormone (ant)agonists in surface and wastewater quality monitoring

Monitoring of chemical water quality is extremely challenging due to the large variety of compounds and the presence of biologically active compounds with unknown chemical identity. Previously, we developed a high resolution Effect-Directed Analysis (EDA) platform that combines liquid chromatography with high resolution mass spectrometry and parallel bioassay detection. In this study, the platform is combined with CALUX bioassays for (anti)androgenic, estrogenic and glucocorticoid activities, and the performance of the platform is evaluated. It appeared to render very repeatable results, with high recoveries of spiked compounds and high consistency between the mass spectrometric and bioassay results. Application of the platform to wastewater treatment plant effluent and surface water samples led to the identification of several compounds contributing to the measured activities. Eventually, a workflow is proposed for the application of the platform in a routine monitoring context. The workflow divides the platform into four phases, of which one to all can be performed depending on the research question and the results obtained. This allows one to make a balance between the effort put into the platform and the certainty and depth by which active compounds will be identified. The EDA platform is a valuable tool to identify unknown bioactive compounds, both in an academic setting as in the context of legislative, governmental or routine monitoring

Sea bass (Dicentrarchus labrax) as a potential bioindicator of estrogenic contamination in marine surface waters

Resumo apresentado sob poster apresentado ao 5th International Symposium on Fish Endocrinology, September, 2004, Castellon, Spain.Exposure of aquatic wildlife in surface waters to (xeno-)estrogens is known to cause reproductive dysfunction. Estrogenic responses in fish are the net result of complex chains of events that will depend on a number of factors, such as bioavailability, bioconcentration/bioaccumulation, and biotransformation. Most of known estrogenic chemicals are lipophilic and hydrophobic and therefore have a strong potential to accumulate in aquatic biota. Therefore, determining environmental exposures may be very difficult and not be particularly meaningful. As test organism the sea bass (Dicentrarchus labrax) was selected, a common species in European marine systems. This work is part of a study focusing on the combination effects of mixtures of estrogenic chemicals in marine and freshwater organisms. Juvenile sea bass were used in order to analyse the bioconcentration and distribution among different tissues of the chemical residues of a set of reference estrogenic chemicals such as 17ß-estradiol (E2), ethynylestradiol (EE2), nonylphenol (NP), octylphenol (OP), bisphenol-A (BPA). Fish were exposed for a period of two weeks to environmentally relevant levels of these compounds, after which liver, bile, muscle, gill and kidney were collected and analyzed. Actual concentrations of E2, EE2 and BPA seawater in the tanks were determined by either gas chromatography with ion trap detection or HPLC coupled to diode array detection. In bile, levels of BPA were determined according to a method presented earlier by Houtman et al. (13th Annual Meeting SETAC Europe, 2003). Actual NP and OP concentrations in both water and tissues were determined by HPLC-ESI-MS according to recently developed methods by Pojana et al. (J. Anal. Chem., in press). Bioconcentration and distribution of residual compounds in tissues were correlated to the levels of plasma vitellogenin (results are presented also at this conference) and to actual exposure concentrations. The general suitability of the sea bass as a bioindicator of estrogenic contamination in the marine environment is discussed.Comissão Europeia (CE) - ACE, EVK1-CT-2001-100