Can mysid shrimp help us unravel possible endocrine disruption in marine environments?

Changes in the hormone regulation in animals due to environmental contaminants (endocrine disruptors) has recently become a widely investigated and politically charged issue. Invertebrates account for 95% of the known species of animals on earth, yet surprisingly little effort has been made to understand their value in signaling potential environmental endocrine disruption. A few reports, however, do suggest that endocrine disruptive effects also affect invertebrates. Due to the high pollutant load of the Scheldt estuary and North Sea, effects on resident populations may occur. A recent database published by our laboratory summarizes the possible effects of potential endocrine disruptive compounds for the North Sea ecosystem. Mysids are used frequently in toxicity studies and there is growing interest in developing toxicity tests with mysids that are indigenous to local ecosystems. Furthermore, United States Environmental Protection Agency (USEPA) has included mysid shrimp as part of a tiered approach in its Endocrine Disruptor Screening Program. In this context, we are examining the potential use of Neomysis integer as a test organism for determining the effects of endocrine disruptors on northern European estuarine communities. The hyperbenthic N. integer dominates the upper regions of European estuaries and are thought to provide a significant link in the exchange of organic matter between the benthic and pelagic systems of estuaries. Cellular and physiological biomarkers for endocrine disruption related to the energy and steroid metabolism of N. integer have been developed and are currently being evaluated in an inter-laboratory validation study. The results obtained with these biomarkers are linked to ecological field observations and measured concentrations of selected endocrine disruptors in the Scheldt estuary (Netherlands). Results of the biomarker studies and field samplings will be highlighted and suggestions on the use of this invertebrate model to test possible endocrine disruptive effects in estuarine environments will be discussed

The testosterone metabolism of <i>Neomysis integer</i>: the quest continues… (poster)

Both vertebrate and invertebrate species use enzymatic biotransformations for detoxication and elimination of xenobiotics. Testosterone has been used as a substrate to study the multiplicity of these enzymes. Since many of these enzymes are under hormone control, disruption of the hormone function can lead to potential effects on enzyme function and subsequently steroid homeostasis. The testosterone metabolism has therefore been proposed as a biomarker of exposure to endocrine disruptive compounds.In a previous study, the estuarine crustacean Neomysis integer (Crustacea, Mysidacea) was exposed to both testosterone and [14C]-testosterone. Identification and quantification of testosterone metabolites and endogenous steroids was done using TLC and LC-MSn (Verslycke et al., Gen. Comp. Endocrinol., accepted). The use of liquid chromatography coupled with multiple mass spectrometry allows a unique quantification of both endogenously produced steroids and in vivo produced metabolites in single mysid. Recent research has focused on the potential use of these biotransformations as a predictive biomarker for exposure to known endocrine disruptors. In this context, quantitative changes in the biotransformation profile of testosterone were evaluated after exposure to tributyltin (TBT), a compound used in antifouling paint, which has been suspected to interfere with steroid metabolism. The resulting protocol allows a quantitative and qualitative evaluation of the effect of TBT on the testosterone metabolism of N. integer. The results of these exposures will be presented and a possible mechanism of disruption through interaction with the P450 enzyme system is proposed.Future research on the steroid metabolism of N. integer could result in the development of predictive biomarkers for detection of endocrine disruption in estuarine environments

Testosterone metabolism in Neomysis integer following exposure to benzo(a)pyrene

Author Posting. © Elsevier B.V., 2006. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 144 (2006): 405-412, doi:10.1016/j.cbpb.2006.04.001.Cytochromes P450 (CYPs) are important enzymes involved in the regulation of hormone synthesis and in the detoxification and/or activation of xenobiotics. CYPs are found in virtually all organisms, from archae, and eubacteria to eukaryota. A number of endocrine disruptors are suspected of exerting their effects through disruption of normal CYP function. Consequently, alterations in steroid hormone metabolism through changes in CYP could provide an important tool to evaluate potential effects of endocrine disruptors. The aim of this study was to investigate the potential effects of the known CYP modulator, benzo(a)pyrene (B(a)P), on the testosterone metabolism in the invertebrate Neomysis integer (Crustacea; Mysidacea). N. integer were exposed for 96h to 0.43, 2.39, 28.83, 339.00 and 1682.86μg B(a)P L-1 and a solventcontrol, and subsequently their ability to metabolize testosterone was assessed. Identification and quantification of the produced phase I and phase II testosterone metabolites was performed using liquid chromatography coupled with multiple mass spectrometry (LC-MS2). Significant changes were observed in the overall ability of N. integer to metabolize testosterone when exposed to 2.39, 28.83, 339.00 and 1682.86μg B(a)P L-1 as compared to the control animals.This research was supported by a research grant of the Ghent University Research Fund (BOF, 011.072.02). Dr. Tim Verslycke was supported by a Postdoctoral Fellowship of the Belgian American Educational Foundation

Mysid crustaceans as potential test organisms for the evaluation of environmental endocrine disruption: a review

Anthropogenic chemicals that disrupt the hormonal systems (endocrine disruptors) of wildlife species recently have become a widely investigated and politically charged issue. Invertebrates account for roughly 95% of all animals, yet surprisingly little effort has been made to understand their value in signaling potential environmental endocrine disruption. This omission largely can be attributed to the high diversity of invertebrates and the shortage of fundamental knowledge of their endocrine systems. Insects and crustaceans are exceptions and, as such, appear to be excellent candidates for evaluating the environmental consequences of chemically induced endocrine disruption. Mysid shrimp (Crustacea: Mysidacea) may serve as a viable surrogate for many crustaceans and have been put forward as suitable test organisms for the evaluation of endocrine disruption by several researchers and regulatory bodies (e.g., the U.S. Environmental Protection Agency). Despite the long-standing use of mysids in toxicity testing, little information exists on their endocrinology, and few studies have focused on the potential of these animals for evaluating the effects of hormone-disrupting compounds. Therefore, the question remains as to whether the current standardized mysid endpoints can be used or adapted to detect endocrine disruption, or if new procedures must be developed, specifically directed at evaluating hormone-regulated endpoints in these animals. This review summarizes the ecological importance of mysids in estuarine and marine ecosystems, their use in toxicity testing and environmental monitoring, and their endocrinology and important hormone-regulated processes to highlight their potential use in assessing environmental endocrine disruption

Mysid crustaceans as standard models for the screening and testing of endocrine-disrupting chemicals

Author Posting. © Springer, 2007. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Ecotoxicology 16 (2007): 205-219, doi:10.1007/s10646-006-0122-0.Investigative efforts into the potential endocrine-disrupting effects of chemicals have mainly concentrated on vertebrates, with significantly less attention paid to understanding potential endocrine disruption in the invertebrates. Given that invertebrates account for at least 95% of all known animal species and are critical to ecosystem structure and function, it remains essential to close this gap in knowledge and research. The lack of progress regarding endocrine disruption in invertebrates is still largely due to: (1) our ignorance of mode-of-action, physiological control, and hormone structure and function in invertebrates; (2) lack of a standardized invertebrate assay; (3) the irrelevance to most invertebrates of the proposed activity-based biological indicators for endocrine disruptor exposure (androgen, estrogen and thyroid); (4) limited field studies. Past and ongoing research efforts using the standard invertebrate toxicity test model, the mysid shrimp, have aimed at addressing some of these issues. The present review serves as an update to a previous publication on the use of mysid shrimp for the evaluation of endocrine disruptors (Verslycke et al., 2004a). It summarizes recent investigative efforts that have significantly advanced our understanding of invertebrate-specific endocrine toxicity, population modeling, field studies, and transgeneration standard test development using the mysid model.Supported by a Fellowship of the Belgian American Educational Foundation

Endocrine disruption in the Scheldt estuary distribution, exposure and effects (ENDIS-RISKS). Final report

ENDIS-RISKS is a multidisciplinary, research project conducted by five institutes. This project aimed to assess the distribution, exposure and effects of endocrine disruptors in the Scheldt estuary, with specific attention to invertebrates. The Scheldt estuary is known to be one of the most polluted estuaries in the world. The industrial areas of Ghent and Antwerp are to a large extent responsible for this pollution. To achieve these goals detailed knowledge of the distribution and long-term effects of these substances is needed. This information is crucial for the development of future-oriented policy measures at the national and European level. The project can be divided into four different research phases. In Phase I the occurance and distribution of endocrine disrupting substances in the Scheldt estuary was studied. Water, sediment, suspended solids and biota were sampled 3 times a year for a period of 4 years (2002-2006). In all these matrices, 7 groups of chemicals were analysed: estrogens, pesticides, phthalates, organotins, polyaromatic components (PCBs, PBDEs), polyaromatic hydrocarbons (PAHs) and phenols. All the analyzed chemicals are on the OSPAR list of priority chemicals or are indicated as endocrine disruptors on this list. The different water samples were also tested using in vitro assays to assess their potential to bind to the (human) estrogen and androgen receptor. Phase II evaluated the exposure of biota occuring in the Scheldt estuary to endocrine disrupting substances. Based on the results of the chemical analysis, priority substances were selected. Phase III studied the effects of endocrine disrupting substances occurring in the Scheldt estuary on resident mysid shrimp populations (laboratory and field studies). Substances of concern were selected and tested in the laboratory to evaluate their effects on the estuarine mysid Neomysis integer. In the context of this project, three new assays using invertebrate-specific endpoints were developed to examine the effect of endocrine disrupting chemicals (EDCs) on molting, embryogenesis and vitellogenesis of N. integer. Finally, in Phase IV laboratory and field results were used to perform a preliminary environmental risk assessment of endocrine disruptors in the Scheldt estuary. Samples were collected along the salinity gradiënt of the Scheldt estuary with the RV Belgica. Water samples were taken with Teflon-coated Go-Flo bottles (10L), sediment samples with Van Veen Grab, biota with a hyperbentic sledge, and suspended particulate matter (SPM) was continuously sampled with an Alfa Laval flow-through centrifuge. For the chemical analysis, protocols were developed to analyse estrogens, organotriazine herbicides, organochlorine pesticides, phtalates, organotins, PAHs, PCBs, and PBDEs in the different matrices: i.e. water, sediment, SPM and biota.Experimental studies were performed to analyse growth, molting, embryogenesis and vitellogenesis of N. integer. These studies were needed to develop ecotoxicological assays to evaluate EDCs on these physiological processes. To study growth of N. integer, organisms were individually transferrred in exposure solutions and molts were collected to measure the growth after each molting. To study embryogenesis, embryos were taking out of the marsupium and placed in multiwell plates. Each day survival, developmental stages and hatching was analysed. To study vitellogenesis, vitellin was isolated from eggs with gelfitration and polyclonal antibodies were developed (in rabbits). With the isolated vitellin and the antibodies an enzyme-linked immunosorbent assay (ELISA) was developed. Vitellin was quatified in ovigerous females exposed to test compound in the laboratory and in females collected from the different sampling sites of the Scheldt estuary. In addition to vitellin levels, energy allocation and testosterone metabolism was examined in field collected mysids. Finally, results from population stu

Pharmaceuticals and personal care products in the environment: What are the big questions?

Background: Over the past 10-15 years, a substantial amount of work has been done by the scientific, regulatory, and business communities to elucidate the effects and risks of pharmaceuticals and personal care products (PPCPs) in the environment. Objective: This review was undertaken to identify key outstanding issues regarding the effects of PPCPs on human and ecological health in order to ensure that future resources will be focused on the most important areas. Data sources: To better understand and manage the risks of PPCPs in the environment, we used the "key question" approach to identify the principle issues that need to be addressed. Initially, questions were solicited from academic, government, and business communities around the world. A list of 101 questions was then discussed at an international expert workshop, and a top-20 list was developed. Following the workshop, workshop attendees ranked the 20 questions by importance. Data synthesis: The top 20 priority questions fell into seven categories: a) prioritization of substances for assessment, b) pathways of exposure, c) bioavailability and uptake, d) effects characterization, e) risk and relative risk, f) antibiotic resistance, and g) risk management. Conclusions: A large body of information is now available on PPCPs in the environment. This exercise prioritized the most critical questions to aid in development of future research programs on the topic.Centro de Investigaciones del Medioambient