RESUMEN
Body mass, fork length, RNA:DNA ratio, specific growth rate, and hepatic EROD activity and CYP1A expression, were measured in three-spined sticklebacks in the River Ray (south west England) at sites downstream of an urban waste water treatment works (WWTW) prior to, and following, remediation of the effluent with granular activated carbon (GAC) tertiary treatment. During the same two-year period fish were also sampled from a neighbouring reference river (R. Ock). The WWTW effluent elevated water temperatures and nutrient content in the R. Ray and rendered a direct comparison of fish populations in the two rivers untenable. Instead, the stability of population parameters within each river during matched pre- and post-remediation periods was compared. Stickleback populations in both rivers were annual but fish in the R. Ray spawned earlier and were larger than those in the R. Ock. In the R. Ray fish gained mass throughout the winter months whereas in the R. Ock growth was much reduced during this period. In fish from the R. Ray the somatic RNA:DNA ratio remained elevated during May-November after remediation, rather than declining as in the same period pre-remediation and as was the case for fish in the R. Ock during both periods. The specific growth rate of the first post-remediation generation of sticklebacks in the R. Ray was higher than that of the previous pre-remediation generation. Following remediation there was no decline in hepatic EROD activity or in the abundance of hepatic CYP1A transcripts in fish in the R. Ray suggesting that the primary route of exposure to contaminants for these fish was not via the water column, and that the change in performance of the fish post-remediation was not impeded by continued exposure to contaminants. Both EROD activity and CYP1A expression increased in fish in the R. Ock during the later stages of the study suggesting that the fish in this river were exposed to an unidentified contaminant episode. This may have been linked with the poorer performance of fish in the R. Ock during the post-remediation period. The improved performance of fish in the R. Ray suggest that there may be factors in good quality secondary treated sewage effluent which can adversely influence the performance of fish populations, directly or indirectly, and which can be removed by tertiary treatment.
Asunto(s)
Tamaño Corporal/efectos de los fármacos , Exposición a Riesgos Ambientales/análisis , Restauración y Remediación Ambiental , Aguas del Alcantarillado/análisis , Smegmamorpha/metabolismo , Contaminantes del Agua/toxicidad , Animales , Biomarcadores/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , ADN/metabolismo , Expresión Génica/efectos de los fármacos , ARN/metabolismo , Ríos/química , Razón de Masculinidad , Smegmamorpha/genética , Eliminación de Residuos Líquidos , Contaminantes del Agua/análisisRESUMEN
A cleanup method was developed to remove coextracted lipids and natural hormones from biota samples in order to test the endocrine-disrupting (ED) capacity of their extracts in in vitro bioassays. Unspiked and spiked fish tissues were cleaned with a combination of dialysis, gel permeation chromatography (GPC), and normal-phase liquid chromatography (NP-HPLC). The spiking mixture consisted of a broad range of environmental pollutants (endocrine disruptors and genotoxic compounds). Chemical recoveries of each test compound, and thyroid-hormone-like and (anti)androgenic activities of the cleaned extracts were investigated. Despite the chemical and toxicological complexity of the spiking mixture and the sequential sample treatment, chemical analysis revealed acceptable recoveries on average: 89 ± 8% after each cleanup step separately and 75 ± 3% after the whole extraction and cleanup procedure in the extracts. In addition, recovered activities in the bioassays were in good agreement with the spiking levels. The developed cleanup method proved to be capable of lipid and natural hormone removal from fish extracts, enabling the measurement of selected endocrine-hormone-like activities in T(4)*-TTR and AR-CALUX bioassays. The method can be used as a sample preparation method of biota samples for toxicity profiling and effect-directed analysis (EDA).
Asunto(s)
Disruptores Endocrinos/toxicidad , Pruebas de Toxicidad/métodos , Contaminantes Químicos del Agua/toxicidad , Animales , Fraccionamiento Químico , Disruptores Endocrinos/química , Disruptores Endocrinos/aislamiento & purificación , Monitoreo del Ambiente/métodos , Peces/metabolismo , Genes Reporteros , Hormonas/química , Hormonas/aislamiento & purificación , Lípidos/química , Lípidos/aislamiento & purificación , Ensayo de Unión Radioligante , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/aislamiento & purificaciónRESUMEN
Effluent discharges at Rodbourne sewage treatment works (STWs) were assessed using chemical and in vitro biological analysis as well as modelling predictions. Results showed that Rodbourne STW discharged less estrone (E1) than expected, but similar 17beta-estradiol (E2) and 17alpha-ethinyl estradiol (EE2) to those predicted by a widely cited effluent prediction model. The Exposure Analysis Modelling System (EXAMS) model was set up using measured effluent concentrations as its starting point to predict estrogen concentrations along a 10 km length of the receiving water of the River Ray. The model adequately simulated estrogen concentrations along the river when compared to July 2007 measured data. The model predicted combined estrogen equivalents in reasonable agreement with estrogenicity as measured by passive sampler (POCIS) extracts using the yeast estrogen screen. Using gauged mean flow values for 2007 the model indicated that the most important determinand for estrogen exposure in the Ray was not season, but proximity to the Rodbourne effluent. Thus, fish in the first 3 km downstream of Rodbourne were typically exposed to two or even three times more estrogens than those living 7-10 km further downstream. The modelling indicated that, assuming the effluent estrogen concentrations measured in February 2008 were typical, throughout the year the whole length of the Ray downstream of Rodbourne would be estrogenic, i.e. exceeding the 1 ng/L E2 equivalent threshold for endocrine disruption.
Asunto(s)
Monitoreo del Ambiente/métodos , Estrógenos/análisis , Modelos Químicos , Ríos/química , Contaminantes Químicos del Agua/análisis , Bioensayo , Estradiol/análisis , Estrona/análisis , Etinilestradiol/análisis , Predicción , Eliminación de Residuos Líquidos , Contaminación Química del Agua/estadística & datos numéricos , Levaduras/genéticaRESUMEN
The assessment of endocrine disrupting potentials of field sediments has until now been mostly limited to classical chemical analysis, in vitro assays and in vivo bioassays performed with vertebrates. There is an urgent need for easy, cheap and reproducible invertebrate tests which may be applied in certain monitoring activities. Since the mudsnail Potamopyrgus antipodarum is known to be tolerant to natural stressors, but also sensitive to endocrine disrupting chemicals, it is very likely that this organism could be suitable for the assessment of endocrine effects of e.g. field sediments. Within this study the endocrine potential of sediments in three European river basins was assessed. The yeast estrogen screen (YES) and a sediment contact test with P. antipodarum were performed. Furthermore, analyses of physico-chemical properties and concentrations of heavy metals, PAHs, organotins, natural steroids and alkylphenols were done. In the sediment contact test, the reproduction of the snail was promoted by a part of the sediments. This phenomenon could not be explained by their physico-chemical properties. However, at some of those sites a high estrogenic activity was detected in the YES, leading to the assumption that endocrine disrupting compounds could be responsible for those effects. This assumption could be confirmed to some extent with partially high concentrations of xeno-estrogens (e.g. nonylphenol) at the certain sites. Our study demonstrates the applicability of the test with P. antipodarum for a variety of sediments and once again points out the need of suitable in vivo biotests for the risk assessment of field sediments.
Asunto(s)
Disruptores Endocrinos/toxicidad , Sedimentos Geológicos/química , Ríos/química , Caracoles/efectos de los fármacos , Animales , Bioensayo , Disruptores Endocrinos/química , Monitoreo del Ambiente , Europa (Continente) , Metales Pesados/química , Metales Pesados/toxicidad , Compuestos Orgánicos de Estaño/química , Compuestos Orgánicos de Estaño/toxicidad , Fenoles/química , Fenoles/toxicidad , Hidrocarburos Policíclicos Aromáticos/química , Hidrocarburos Policíclicos Aromáticos/toxicidad , Contaminantes del Suelo/química , Contaminantes del Suelo/toxicidad , Esteroides/química , Esteroides/toxicidadRESUMEN
This paper presents a study evaluating the suitability of recombinant yeast-based estrogenicity assays as a pre-screening tool for monitoring of the chemical status of water bodies in support of the Water Framework Directive (WFD). Three different recombinant yeast-based assays were evaluated; the Yeast Estrogen Screen (YES), the Recombinant Yeast Assay (RYA) and the Rikilt Estrogen bioAssay (REA), of which the YES assay was employed by two different laboratories. No significant difference between the performance of neither the different laboratories, nor the different yeast-assays was observed. Six batches of eleven samples each were analysed one week apart by the four participating laboratories and the robustness, repeatability and reproducibility of the participating yeast-based assays were evaluated. The setup included a correlation between bioassay results and results from chemical target analysis, which gave valuable information in the evaluation of the assays' performance. A good agreement was found between chemical and bioassay results, showing that the yeast-based assays can give valuable information in WFD work. However, the low sensitivity of the assays towards alkylphenols needs to be significantly improved if they are to be used for monitoring of these compounds. The study further led to suggestions on ways to improve traceability and quality assurance of the yeast-based assays.
Asunto(s)
Estrógenos/análisis , Tamizaje Masivo/métodos , Contaminantes Químicos del Agua/análisis , Levaduras/efectos de los fármacos , Bioensayo/métodos , Sensibilidad y Especificidad , Levaduras/genéticaRESUMEN
Produced water collected from oil and gas platforms on the United Kingdom Continental Shelf was characterized for nonregulated pollutants through an effects-directed analysis procedure. Produced water samples were characterized for the presence of aryl hydrocarbon receptor (AhR) and estrogen receptor (ER) agonists using the dioxin-responsive, chemical-activated luciferase gene expression assay (DR-CALUX) and yeast estrogen screen (YES) bioassays. The AhR and ER agonists were then isolated by normal-phase, high-performance liquid chromatography and identified using gas chromatography coupled to mass spectrometry in a number of formats. The identified compounds were cross-referenced with those compounds routinely analyzed and regarded by the Oslo and Paris (OSPAR) Commission for the North East Atlantic as priority hazardous substances. The occurrence in produced water of a number of nonregulated compounds with demonstrable potential environmental effects is presented, to our knowledge for the first time. These include persistent organic contaminants, such as hexachlorobenzene, decachlorobiphenyl, and octachlorodibenzofuran.
Asunto(s)
Agua de Mar/química , Contaminantes Químicos del Agua/análisis , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas , Expresión Génica , Luciferasas/genética , Receptores de Hidrocarburo de Aril/agonistas , Receptores de Estrógenos/agonistasRESUMEN
The characterisation of estrogen (ER) and arylhydrocarbon (AhR) receptor agonists was performed in extracts of the tissues of transplanted blue mussels (Mytilus edulis). The yeast estrogen screen (YES) was used to detect the presence of ER agonists, whilst the DR-CALUX assay was used to detect AhR agonists. The concentration of ER agonists in mussel tissue from the Tees estuary, Brancaster and River Crouch (UK) were below the limits of detection for the YES assay (0.87 pg E2 g(-1)). AhR agonists were measured at concentrations of between 1 and 950 pg TCDD g(-1) in mussel tissue. A bioassay-directed fractionation of the sample extracts, followed by advanced broad spectrum gas chromatography-mass spectrometry analysis was then used in an attempt to identify the AhR agonists present. This showed that a complex mixture of AhR agonists occurs in the samples and that further work will be required in order to isolate and identify the individual compounds responsible.
Asunto(s)
Estrógenos/agonistas , Mytilus edulis/metabolismo , Receptores de Hidrocarburo de Aril/agonistas , Contaminantes Químicos del Agua/análisis , Animales , Bioensayo , Monitoreo del Ambiente , Estrógenos/análisis , Cromatografía de Gases y Espectrometría de Masas , Receptores de Hidrocarburo de Aril/análisis , Ríos/química , Agua de Mar/química , Extractos de Tejidos/análisis , Contaminantes Químicos del Agua/toxicidadRESUMEN
The in vitro aryl hydrocarbon receptor (AhR) agonist potency of offshore produced water effluents, collected from the United Kingdom Continental Shelf, was determined using the dioxin responsive (DR)-chemically activated luciferase expression (CALUX) assay. Octadecylsilane (C18) solid phase extraction (SPE) extracts of produced water were exposed to DR-CALUX cells for 24h in order to investigate the contribution in potency from compounds that are stable to metabolism by the CALUX cells during exposure. The stable AhR agonist potency determined over 24h was highly variable and ranged from 1 to 430 ng TCDD TEQ(CALUX)l(-1). These data reflect the highly variable composition of produced water discharges from different production fields. It is recommended that further work be performed to characterise the full range of stable dioxin like AhR agonists present in offshore produced water discharges using techniques such as bioassay-directed analysis.
Asunto(s)
Dioxinas/análisis , Receptores de Hidrocarburo de Aril/agonistas , Contaminantes Químicos del Agua/análisis , Agua/química , Calibración , Luciferasas , Dibenzodioxinas Policloradas/química , Pruebas de Toxicidad/métodos , Reino Unido , Eliminación de Residuos LíquidosRESUMEN
The DR-CALUX assay has been utilised for the bio-analytical screening of a number of estuarine sediments for dioxin-like activity. Total sediment extracts (samples containing all extracted compounds) and cleaned-up extracts (samples with the most stable compounds isolated from the total extracts) were screened. The concentration of the stable dioxin-like compounds in the cleaned-up sediment extracts was between 1.0 and 106 pgTEQCALUX g(-1) dry weight. The majority of sediments contained levels of dioxin-like compounds that were above concentrations that are considered to be a low risk to aquatic organisms. The CALUX bio-analytical approach showed some disparity with the traditional analytical approach. The reasons for these differences have been identified tentatively: firstly, the DR-CALUX assay responds to all dioxin-like compounds, and secondly, it measures non-additive effects. The dioxin-like activity of compounds in sediment total extracts, which contain both labile and stable compounds, were also assessed and were six orders of magnitude higher than the cleaned-up samples. This suggests the vast majority of the total dioxin-like activity is attributable to labile compounds. Overall, the DR-CALUX assay is shown to be a useful tool in the assessment of dioxin-like activity in estuarine sediments.
Asunto(s)
Dioxinas/análisis , Sedimentos Geológicos/química , Luciferasas de Luciérnaga/biosíntesis , Contaminantes Químicos del Agua/análisis , Animales , Bioensayo/métodos , Carcinoma Hepatocelular , Monitoreo del Ambiente/métodos , Neoplasias Hepáticas , Luciferasas de Luciérnaga/genética , Ratas , Transfección , Células Tumorales Cultivadas , Reino UnidoRESUMEN
The in vitro estrogen receptor (ER) agonist potency and C1 to C9 alkyl substituted phenol content of offshore produced water effluents collected from the UK sector of the North Sea were determined using a combination of bio-analytical and chemical analysis techniques. An in vitro reporter gene assay was used to determine ER agonist potency, whilst gas chromatography coupled to mass spectrometry (GC-MS) was used to quantify the concentration of alkylphenols. The in vitro ER agonist potency was highly variable and ranged from less than the limit of detection (theoretically 0.03 ng 17beta-estradiol (E2) l(-1)) to 91 ng E2 l(-1). C1 to C5 alkylphenol concentrations were also highly variable ranging from 5 to 1600 microg l(-1) with a median concentration of 206 microg l(-1). These data reflect the highly variable composition of produced water discharges from different fields. The observed poor correlation of the alkylphenol isomer content and ER agonist activity suggests that other compounds present in the produced water discharges may be responsible for the ER agonist activity observed. It is recommended that further work be performed to characterise the full range of ER agonists present in offshore produced water discharges.
Asunto(s)
Monitoreo del Ambiente/métodos , Fenoles/análisis , Receptores de Estrógenos/agonistas , Contaminantes Químicos del Agua/análisis , Cromatografía de Gases y Espectrometría de Masas , Mar del Norte , Valores de ReferenciaRESUMEN
The estrogen receptor (ER) agonist potency of offshore produced water discharges was examined via bioassay-directed chemical analysis. The in vitro estrogen receptor (ER) and androgen receptor (AR) agonist potency of five produced water samples collected from oil-production platforms in the British and Norwegian sectors of the North Sea was determined by using the yeast estrogen and androgen screens. Produced water samples were extracted in situ on the production platforms by using large-volume solid-phase extraction. All five extracts tested positive for the presence of ER agonists, whereas no AR agonist activity could be detected. By using the yeast estrogen screen assay in association with bioassay-directed fractionation, attempts were made to identify the ER agonist compounds present in the produced water extracts. The fractionation procedure used cyano-amino-bonded silica normal-phase high-performance liquid chromatography to isolate estrogenic compounds from produced water extract followed by full-scan gas chromatography-electron-impact mass spectrometry (GC-(EI)MS) to identify them. Isomeric mixtures of C1 to C5 and C9 alkylphenols contributed to the majority of the ER agonist potency measured in the samples.
Asunto(s)
Andrógenos , Receptores de Estrógenos/agonistas , Agua de Mar/análisis , Bioensayo/métodos , Técnicas de Química Analítica/métodos , Cromatografía Líquida de Alta Presión , Monitoreo del Ambiente , Cromatografía de Gases y Espectrometría de Masas , Mar del Norte , Fenoles/análisis , Fenoles/química , Agua de Mar/químicaRESUMEN
The activity of estrogen-receptor (ER) agonists in sediments collected from the United Kingdom (UK) estuaries was assessed using the in vitro recombinant yeast estrogen screen (YES assay). The YES assay was successfully used to determine the in vitro ER agonist potency of pore waters and solvent extracts of sediments collected from UK estuaries. Estrogen-receptor agonists were detected in 66% of the pore water samples and in 91% of the sediment solvent extracts tested. The pore waters tested had ER agonist potencies from less than 2 to 68 ng 17beta-estradiol (E2) L(-1), whereas sediment extracts had potencies from less than 0.2 to 13 microg E2 kg(-1). A toxicity identification evaluation approach using bioassay-directed fractionation was used in an attempt to identify the ER agonists in extracts of sediments collected from the Tyne and Tees estuaries (UK). Gas chromatography-mass spectrometry was used to provide lists of compounds in the fractions obtained that were evaluated for known ER agonist activity using published data and an ER quantitative structure-activity relationship model. Toxicity identification evaluation characterization failed to identify any ER agonists in pore water extracts; however, three compounds in sediment solvent extracts were identified as ER agonists. Nonylphenol, cinnarizine, and cholesta-4,6-dien-3-one were identified in the sample collected from the Tyne estuary. Important ER agonist substances that contaminate marine sediments remain unidentified. The present study as well as previous work on effluents point toward the involvement of natural products in the estrogenic burdens of marine sediments. Further work is required to establish the relative contribution of natural products and anthropogenic chemicals to current environmental impacts in the context of the Oslo and Paris Commission strategy to eliminate hazardous substances by 2020.
Asunto(s)
Sedimentos Geológicos/análisis , Modelos Químicos , Relación Estructura-Actividad Cuantitativa , Receptores de Estrógenos/agonistas , Agua/análisis , Levaduras/metabolismo , Bioensayo , Cromatografía Líquida de Alta Presión , Contaminación Ambiental/prevención & control , Cromatografía de Gases y Espectrometría de Masas , Agua de Mar , Reino Unido , Levaduras/efectos de los fármacosRESUMEN
The mutagenic activity bioassay Mutatox was used to assess the mutagenic activity associated with sediments collected from five UK estuaries. Assays were performed on extracts of sediment pore water and residual particulate material isolated from sediment samples collected from the rivers Tyne, Tees, Mersey, and Thames as well as Southampton Water. No mutagenic activity was associated with the pore water extracts, however, 7 of the 28 organic solvent extracts of sediment particulate material contained potential genotoxins. By using Mutatox in association with bioassay-directed fractionation, attempts were made to identify the mutagenic compounds present in the extracts. The fractionation procedure used normal phase solid phase extraction, C18 reverse phase HPLC and cyano/amino bonded silica normal phase HPLC. GC-MS (EI and NICI) analysis was used to identify polycyclic aromatic hydrocarbons (PAH), alkyl substituted PAH, nitro-polycyclic aromatic compounds (nitro-PACs), polycyclic aromatic ketones, oxygenated-PACs, and other known mutagens contributing to the genotoxicity measured in the samples. Some potentially genotoxic compounds remain unidentified.