Synthetic glucocorticoids in the aquatic environment: their potential impacts on fish

This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University.Human pharmaceuticals have been shown to be entering the aquatic environment in quantities sufficient to produce adverse effects to aquatic organisms, particularly fish. The impacts of synthetic oestrogens have been well documented, but other groups of steroidal pharmaceuticals have not yet been studied. Hence, the present research was designed to study synthetic glucocorticoids (GCs), which are used in large amounts as immunosuppressive and anti-inflammatory drugs. This study involved different approaches, including in silico, in vitro, in vivo and genomics, to assess the effects of GCs on fish. Using reliable data on consumption of GCs in the UK and the LF2000-WQX hydrological model, mean concentrations of GCs in the river Thames were predicted to be in the range from 30 ng/L to 850 ng/L. Mammalian cell lines were transiently transfected with trout corticosteroid receptors (GR1, GR2 and MR) and the transactivation abilities of ten of the most prescribed GCs in the UK were measured in vitro. All tested GCs showed significantly higher activity with GR2 than with GR1. In order to assess the impact of low concentrations of GCs in vivo, two chronic exposure experiments were conducted with adult fathead minnows (Pimephales promelas). Both experiments showed potency-related and concentration-related impacts on various endpoints. There was a concentration-related increase in plasma glucose concentrations and a decrease in blood lymphocyte count. Induction of secondary sexual characters in females suggests a concentration-related masculinisation of fathead minnows. There was a decreasing trend in plasma vitellogenin concentrations in female fish with increasing exposure concentration of GCs. Expression profiles of selected genes (PEPCK, GR and Vtg) in liver also demonstrated concentration-related effects at all three tested concentrations. Hence, it was not possible to define a no effect concentration for the tested GCs. This study probably provides reliable estimates of the likely range of concentrations of GCs in a typical river, impacted by effluent from many sewage treatment plants. The in vitro results indicate that all tested GCs bind to fish GR in a similar manner to that reported for mammalian receptors. The in vivo results suggest that GCs could cause effects at very low (as low as 100 ng/L) concentrations that could be environmentally-relevant. The immunosuppresive effects could make fish susceptible to disease and the reproductive effects may have population-level impacts. It is very likely that the effects of different GCs will be additive, as has been shown for oestrogenic chemicals. Therefore, this study warrants further environmental risk assessment of GCs, especially in mixture scenarios.Commonwealth Scholarship Commissio

Mixture effects at very low doses with combinations of anti-androgenic pesticides, antioxidants, industrial pollutant and chemicals used in personal care products

This article has been made available through the Brunel Open Access Publishing Fund.Many xenobiotics have been identified as in vitro androgen receptor (AR) antagonists, but information about their ability to produce combined effects at low concentrations ismissing. Such data can reveal whether joint effects at the receptor are induced at low levels andmay support the prioritisation of in vivo evaluations and provide orientations for the grouping of anti-androgens in cumulative risk assessment. Combinations of 30 AR antagonists from a wide range of sources and exposure routes (pesticides, antioxidants, parabens, UV-filters, synthetic musks, bisphenol-A, benzo(a)pyrene, perfluorooctane sulfonate and pentabromodiphenyl ether) were tested using a reporter gene assay (MDA-kb2). Chemicalswere combined at threemixture ratios, equivalent to single components' effect concentrations that inhibit the action of dihydrotesterone by 1%, 10% or 20%. Concentration addition (CA) and independent action were used to calculate additivity expectations. We observed complete suppression of dihydrotestosterone effects when chemicals were combined at individual concentrations eliciting 1%, 10% or 20% AR antagonistic effect. Due to the large number of mixture components, the combined AR antagonistic effects occurred at very low concentrations of individual mixture components. CA slightly underestimated the combined effects at all mixture ratios. In conclusion, large numbers of AR antagonists froma wide variety of sources and exposure routes have the ability of acting together at the receptor to produce joint effects at very low concentrations. Significant mixture effects are observed when chemicals are combined at concentrations that individually do not induce observable AR antagonistic effects. Cumulative risk assessment for AR antagonists should apply grouping criteria based on effects where data are available, rather than on criteria of chemical similarity

Synthetic glucocorticoids in the aquatic environment : their potential impacts on fish

Human pharmaceuticals have been shown to be entering the aquatic environment in quantities sufficient to produce adverse effects to aquatic organisms, particularly fish. The impacts of synthetic oestrogens have been well documented, but other groups of steroidal pharmaceuticals have not yet been studied. Hence, the present research was designed to study synthetic glucocorticoids (GCs), which are used in large amounts as immunosuppressive and anti-inflammatory drugs. This study involved different approaches, including in silico, in vitro, in vivo and genomics, to assess the effects of GCs on fish. Using reliable data on consumption of GCs in the UK and the LF2000-WQX hydrological model, mean concentrations of GCs in the river Thames were predicted to be in the range from 30 ng/L to 850 ng/L. Mammalian cell lines were transiently transfected with trout corticosteroid receptors (GR1, GR2 and MR) and the transactivation abilities of ten of the most prescribed GCs in the UK were measured in vitro. All tested GCs showed significantly higher activity with GR2 than with GR1. In order to assess the impact of low concentrations of GCs in vivo, two chronic exposure experiments were conducted with adult fathead minnows (Pimephales promelas). Both experiments showed potency-related and concentration-related impacts on various endpoints. There was a concentration-related increase in plasma glucose concentrations and a decrease in blood lymphocyte count. Induction of secondary sexual characters in females suggests a concentration-related masculinisation of fathead minnows. There was a decreasing trend in plasma vitellogenin concentrations in female fish with increasing exposure concentration of GCs. Expression profiles of selected genes (PEPCK, GR and Vtg) in liver also demonstrated concentration-related effects at all three tested concentrations. Hence, it was not possible to define a no effect concentration for the tested GCs. This study probably provides reliable estimates of the likely range of concentrations of GCs in a typical river, impacted by effluent from many sewage treatment plants. The in vitro results indicate that all tested GCs bind to fish GR in a similar manner to that reported for mammalian receptors. The in vivo results suggest that GCs could cause effects at very low (as low as 100 ng/L) concentrations that could be environmentally-relevant. The immunosuppresive effects could make fish susceptible to disease and the reproductive effects may have population-level impacts. It is very likely that the effects of different GCs will be additive, as has been shown for oestrogenic chemicals. Therefore, this study warrants further environmental risk assessment of GCs, especially in mixture scenarios.EThOS - Electronic Theses Online ServiceCommonwealth Scholarship CommissionGBUnited Kingdo

Effects of common pesticides on prostaglandin D2 (PGD2) inhibition in SC5 mouse sertoli cells, evidence of binding at the cox-2 active site, and implications for endocrine disruption

Background: There are concerns that diminished prostaglandin action in fetal life could increase the risk of congenital malformations. Many endocrine-disrupting chemicals have been found to suppress prostaglandin synthesis, but to our knowledge, pesticides have never been tested for these effects. Objectives: We assessed the ability of pesticides that are commonly used in the European Union to suppress prostaglandin D2 (PGD2) synthesis. Methods: Changes in PGD2 secretion in juvenile mouse Sertoli cells (SC5 cells) were measured using an ELISA. Coincubation with arachidonic acid (AA) was conducted to determine the site of action in the PGD2 synthetic pathway. Molecular modeling studies were performed to assess whether pesticides identified as PGD2-active could serve as ligands of the cyclooxygenase-2 (COX-2) binding pocket. Results: The pesticides boscalid, chlorpropham, cypermethrin, cyprodinil, fenhexamid, fludioxonil, imazalil (enilconazole), imidacloprid, iprodione, linuron, methiocarb, o-phenylphenol, pirimiphos- methyl, pyrimethanil, and tebuconazole suppressed PGD2 production. Strikingly, some of these substances—o-phenylphenol, cypermethrin, cyprodinil, linuron, and imazalil (enilconazole)— showed potencies (IC50) in the range between 175 and 1,500 nM, similar to those of analgesics intended to block COX enzymes. Supplementation with AA failed to reverse this effect, suggesting that the sites of action of these pesticides are COX enzymes. The molecular modeling studies revealed that the COX-2 binding pocket can accommodate most of the pesticides shown to suppress PGD2 synthesis. Some of these pesticides are also capable of antagonizing the androgen receptor. Conclusions: Chemicals with structural features more varied than previously thought can suppress PGD2 synthesis. Our findings signal a need for in vivo studies to establish the extent of endocrinedisrupting effects that might arise from simultaneous interference with PGD2 signaling and androgen action

Internal exposure dynamics drive the Adverse Outcome Pathways of synthetic glucocorticoids in fish

The Adverse Outcome Pathway (AOP) framework represents a valuable conceptual tool to systematically integrate existing toxicological knowledge from a mechanistic perspective to facilitate predictions of chemical-induced effects across species. However, its application for decision-making requires the transition from qualitative to quantitative AOP (qAOP). Here we used a fish model and the synthetic glucocorticoid beclomethasone dipropionate (BDP) to investigate the role of chemical-specific properties, pharmacokinetics, and internal exposure dynamics in the development of qAOPs. We generated a qAOP network based on drug plasma concentrations and focused on immunodepression, skin androgenisation, disruption of gluconeogenesis and reproductive performance. We showed that internal exposure dynamics and chemical-specific properties influence the development of qAOPs and their predictive power. Comparing the effects of two different glucocorticoids, we highlight how relatively similar in vitro hazard-based indicators can lead to different in vivo risk. This discrepancy can be predicted by their different uptake potential, pharmacokinetic (PK) and pharmacodynamic (PD) profiles. We recommend that the development phase of qAOPs should include the application of species-species uptake and physiologically-based PK/PD models. This integration will significantly enhance the predictive power, enabling a more accurate assessment of the risk and the reliable transferability of qAOPs across chemicals.This work was funded by a Biotechnology and Biological Sciences Research Council (BBSRC) Research Grant (BB/100646X/1), co-funded by the AstraZeneca Global Safety, Health and Environment research programme, to JPS and MR-W supporting LM-C

The read-across hypothesis and environmental risk assessment of pharmaceuticals

This article is made available through the Brunel Open Access Publishing Fund. Copyright © 2013 American Chemical Society.Pharmaceuticals in the environment have received increased attention over the past decade, as they are ubiquitous in rivers and waterways. Concentrations are in sub-ng to low μg/L, well below acute toxic levels, but there are uncertainties regarding the effects of chronic exposures and there is a need to prioritise which pharmaceuticals may be of concern. The read-across hypothesis stipulates that a drug will have an effect in non-target organisms only if the molecular targets such as receptors and enzymes have been conserved, resulting in a (specific) pharmacological effect only if plasma concentrations are similar to human therapeutic concentrations. If this holds true for different classes of pharmaceuticals, it should be possible to predict the potential environmental impact from information obtained during the drug development process. This paper critically reviews the evidence for read-across, and finds that few studies include plasma concentrations and mode of action based effects. Thus, despite a large number of apparently relevant papers and a general acceptance of the hypothesis, there is an absence of documented evidence. There is a need for large-scale studies to generate robust data for testing the read-across hypothesis and developing predictive models, the only feasible approach to protecting the environment.BBSRC Industrial Partnership Award BB/ I00646X/1 and BBSRC Industrial CASE Partnership Studentship BB/I53257X/1 with AstraZeneca Safety Health and Environment Research Programme

Future water quality monitoring - Adapting tools to deal with mixtures of pollutants in water resource management

Environmental quality monitoring of water resources is challenged with providing the basis for safeguarding the environment against adverse biological effects of anthropogenic chemical contamination from diffuse and point sources. While current regulatory efforts focus on monitoring and assessing a few legacy chemicals, many more anthropogenic chemicals can be detected simultaneously in our aquatic resources. However, exposure to chemical mixtures does not necessarily translate into adverse biological effects nor clearly shows whether mitigation measures are needed. Thus, the question which mixtures are present and which have associated combined effects becomes central for defining adequate monitoring and assessment strategies. Here we describe the vision of the international, EU-funded project SOLUTIONS, where three routes are explored to link the occurrence of chemical mixtures at specific sites to the assessment of adverse biological combination effects. First of all, multi-residue target and non-target screening techniques covering a broader range of anticipated chemicals co-occurring in the environment are being developed. By improving sensitivity and detection limits for known bioactive compounds of concern, new analytical chemistry data for multiple components can be obtained and used to characterise priority mixtures. This information on chemical occurrence will be used to predict mixture toxicity and to derive combined effect estimates suitable for advancing environmental quality standards. Secondly, bioanalytical tools will be explored to provide aggregate bioactivity measures integrating all components that produce common (adverse) outcomes even for mixtures of varying compositions. The ambition is to provide comprehensive arrays of effect-based tools and trait-based field observations that link multiple chemical exposures to various environmental protection goals more directly and to provide improved in situ observations for impact assessment of mixtures. Thirdly, effect-directed analysis (EDA) will be applied to identify major drivers of mixture toxicity. Refinements of EDA include the use of statistical approaches with monitoring information for guidance of experimental EDA studies. These three approaches will be explored using case studies at the Danube and Rhine river basins as well as rivers of the Iberian Peninsula. The synthesis of findings will be organised to provide guidance for future solution-oriented environmental monitoring and explore more systematic ways to assess mixture exposures and combination effects in future water quality monitoring.Seventh Framework Programme (E.U

Impact of post - tsunami rehabilitation activities on selective fishing in the Northern coast of Jaffna peninsula

A questionnaire based survey was eonducted in May 2006, among the Tsunami affected fishermen of'Northern Jaffna peninsula, in order to assess the fishing effort after Tsunami rehabilitation activities.4337 families along the coast from Valvettithurai to Kaddaikadu were included in to the survey andthey were divided in to 72 sites according to their respective donor agencies. 16 Non governmentalorganizations were identified as donor agencies for permanent housing schemes, out of which 6agencies directly donated fishing gears and boats. The types and numbers offishing gears and boatsused before and after tsunami were recorded.Results indicate that all the traditional canoes have been replaced by fiber reinforced boats and thenumber of boats showed 12% increase after tsunami. Before tsunami, there were diverse types offishing gears with wide range of mesh size. But post tsunami rehabil itation aids provided on Iy the gi IInets of2 different mesh sizes, one for fish and the other for skates. Although large numbers of boatshave been given, most of them are not in use because of the prevailing unrest situation in this region.Therefore fishing effort due to the increased number of boats is immaterial. But reduced diversity offishing gear will have major impact on stock. Since fishing is restricted in to a narrow belt along thecoast repeated use of skate gill net by most of the fishermen will lead to an over exploitation of skatespecies. Similarly, selective fishing by gill net will deplete coastal fish species. Therefore donor agenciesshould focus on remedial actions in order to diversify the fishing gears in type and in mesh size