RESUMO
The masking of specific effects in in vitro assays by cytotoxicity is a commonly known phenomenon. This may result in a partial or complete loss of effect signals. For common in vitro assays, approaches for identifying and quantifying cytotoxic masking are partly available. However, a quantification of cytotoxicity-affected signals is not possible. As an alternative, planar bioassays that combine high-performance thin layer chromatography with in vitro assays, such as the planar yeast estrogen screen (p-YES), might allow for a quantification of cytotoxically affected signals. Affected signals form a typical ring structure with a supressed or completely lacking centre that results in a double peak chromatogram. This study investigates whether these double peaks can be used for fitting a peak function to extrapolate the theoretical, unaffected signals. The precision of the modelling was evaluated for four individual peak functions, using 42 ideal, undistorted peaks from estrogenic model compounds in the p-YES. Modelled ED50-values from bisphenol A (BPA) experiments with cytotoxically disturbed signals were 13 times higher than for the apparent data without compensation for cytotoxicity (320 ± 63 ng versus 24 ± 17 ng). This finding has a high relevance for the modelling of mixture effects according to concentration addition that requires unaffected, complete dose-response relationships. Finally, we applied the approach to results of a p-YES assay on leachate samples of an elastomer material used in water engineering. In summary, the fitting approach enables the quantitative evaluation of cytotoxically affected signals in planar in vitro assays and also has applications for other fields of chemical analysis like distorted chromatography signals.
Assuntos
Bioensaio , Bioensaio/métodos , Cromatografia em Camada Fina/métodos , Fenóis/toxicidade , Fenóis/análise , Fenóis/química , Compostos Benzidrílicos/toxicidade , Compostos Benzidrílicos/análise , Compostos Benzidrílicos/química , Estrogênios/análise , Estrogênios/toxicidadeRESUMO
Over the last two decades, effect-directed analysis (EDA) gained importance as a seminal screening tool for tracking biological effects of environmental organic micro-pollutants (MPs). As EDA using high-performance liquid chromatography and bioassays is costly and time consuming, recent implementations of this approach have combined high-performance thin-layer chromatography (HPTLC) with effect-based methods (EBMs) using cell-based bioassays, enabling the detection of estrogenic, androgenic, genotoxic, photosystem II (PSII)- inhibiting, and dioxin-like sample components on a HPTLC plate. In the present study, the developed methodologies were applied as a HPTLC-based bioassay battery, to investigate toxicant elimination efficiency of wastewater treatment plants (WWTPs), and to characterize the toxic potential of landfill leachates. Activity levels detected in untreated landfill leachates, expressed as reference compound equivalence (EQ) concentration, were up to 16.8 µg ß-naphthoflavone-EQ L-1 (indicating the degree of dioxin-like activity), 1.9 µg estradiol-EQ L-1 (estrogenicity) and 8.3 µg diuron-EQ L1 (PSII-inhibition), dropping to maximal concentrations of 47 ng ß-naphthoflavone-EQ L-1, 0.7 µg estradiol-EQ L-1 and 53.1 ng diuron-EQ L-1 following treatment. Bisphenol A (BPA) is suggested to be the main contributor to estrogenic activity, with concentrations determined by the planar yeast estrogen screen corresponding well to results from chemical analysis. In the investigated WWTP samples, a decrease of estrogenic activity of 6-100% was observed following treatment for most of the active fractions, except of a 20% increase in one fraction (Rf = 0.568). In contrast, androgenicity with concentrations up to 640 ng dihydrotestosterone-EQ L-1 was completely removed by treatment. Interestingly, genotoxic activity increased over the WWTP processes, releasing genotoxic fractions into receiving waters. We propose this combined HPTLC and EBM battery to contribute to an efficient, cheap, fast and robust screening of environmental samples; such an assay panel would allow to gain an estimate of potential biological effects for prioritization prior to substance identification, and its routine application will support an inexpensive identification of the toxicity drivers as a first tier in an EDA strategy.
Assuntos
Bioensaio/métodos , Poluentes Químicos da Água/toxicidade , Purificação da Água , Compostos Benzidrílicos , Cromatografia em Camada Fina/métodos , Monitoramento Ambiental/métodos , Estrogênios/toxicidade , Fenóis , Dibenzodioxinas Policloradas/análise , Águas Residuárias/análise , beta-NaftoflavonaRESUMO
We present a novel tool for detecting and monitoring photosystem II (PSII) inhibitors, using the freshwater alga Desmodesmus subspicatus, in environmental samples fractionated by high-performance thin-layer chromatography (HPTLC). After chromatographic separation of a sample on a HPTLC plate, the algal suspension is sprayed homogeneously on the plate, and PSII-inhibition by specific sample components is detected based on changes in fluorescence yield, viewed by a maxi Imaging-Pulse-Amplitude-Modulation fluorometer. Dose-dependent responses to the PSII-inhibitor herbicides atrazine and diuron, frequently detected in water bodies, are demonstrated without and with chromatographic separation. The limits of quantification for atrazine and diuron with chromatographic separation were 1.94 ng and 99 pg, respectively, allowing the detection of environmentally relevant concentrations of these herbicides. The developed method was also employed to analyze sample extracts collected during a passive sampling campaign in surface waters. The obtained data correlated well with results from LC-MS/MS chemical analysis but also revealed unknown PSII-inhibiting activities. The proposed methodology represents a rapid and sensitive screening tool for the simultaneous effect-based detection of PSII-inhibitors in environmental samples.
Assuntos
Herbicidas , Poluentes Químicos da Água , Cromatografia Líquida , Cromatografia em Camada Fina , Água Doce , Complexo de Proteína do Fotossistema II , Espectrometria de Massas em TandemRESUMO
We present an innovative technological platform for monitoring the direct genotoxicity of individual components in complex environmental samples, based on bioluminescent Escherichia coli genotoxicity bioreporters, sprayed onto the surface of a high-performance thin-layer chromatography (HPTLC) plate. These sensor strains harbor plasmid-borne fusions of selected gene promoters of the E. coli SOS DNA repair system to the Photorhabdus luminescens luxABCDE gene cassette, and mark by increased luminescence the presence of potentially DNA-damaging sample components separated on the plate. We demonstrate an "on plate" quantifiable dose-dependent response to several model genotoxicants (without metabolic activation). We further demonstrate the applicability of the system by identifying as genotoxic specific components of HPTLC-separated influent and effluent samples of wastewater treatment plants, thereby alleviating the need for a comprehensive chemical analysis of the sample.
Assuntos
Escherichia coli , Photorhabdus , Cromatografia em Camada Fina , Dano ao DNA , PlasmídeosRESUMO
The potential to bioconcentrate is generally considered to be an unwanted property of a substance. Consequently, chemical legislation, including the European REACH regulations, requires the chemical industry to provide bioconcentration data for chemicals that are produced or imported at volumes exceeding 100 tons per annum or if there is a concern that a substance is persistent, bioaccumulative, and toxic. For the filling of the existing data gap for chemicals produced or imported at levels that are below this stipulated volume, without the need for additional animal experiments, physiologically-based toxicokinetic (PBTK) models can be used to predict whole-body and tissue concentrations of neutral organic chemicals in fish. PBTK models have been developed for many different fish species with promising results. In this study, we developed PBTK models for zebrafish (Danio rerio) and roach (Rutilus rutilus) and combined them with existing models for rainbow trout (Onchorhynchus mykiss), lake trout (Salvelinus namaycush), and fathead minnow (Pimephales promelas). The resulting multispecies model framework allows for cross-species extrapolation of the bioaccumulative potential of neutral organic compounds. Predictions were compared with experimental data and were accurate for most substances. Our model can be used for probabilistic risk assessment of chemical bioaccumulation, with particular emphasis on cross-species evaluations.
Assuntos
Peixes , Modelos Biológicos , Toxicocinética , Poluentes Químicos da Água/farmacocinética , Animais , Compostos Orgânicos , Poluentes Químicos da Água/química , Poluentes Químicos da Água/toxicidadeRESUMO
We report the results of a study of the potential negative effects of the freeze-drying process, normally considered a benign means for long-term conservation of living cells and the golden standard in bacterial preservation. By monitoring gene induction using a whole-cell Escherichia coli bioreporter panel, in which diverse stress-responsive gene promoters are fused to luminescent or fluorescent reporting systems, we have demonstrated that DNA repair genes belonging to the SOS operon (recA, sulA, uvrA, umuD, and lexA) were induced upon resuscitation from the freeze-dried state, whereas other stress-responsive promoters such as grpE, katG, phoA, soxS, and sodA were not affected. This observation was confirmed by the UMU-chromotest (activation of the umuD gene promoter) in Salmonella typhimurium, as well as by real-time PCR analyses of selected E. coli SOS genes. We further show that a functional SOS operon is important in viability maintenance following resuscitation, but that at the same time, this repair system may introduce significantly higher mutation rates, comparable to those induced by high concentrations of a known mutagen. Our results also indicate that the entire freeze-drying process, rather than either freezing or drying separately, is instrumental in the induction of DNA damage.
Assuntos
Escherichia coli/fisiologia , Escherichia coli/efeitos da radiação , Liofilização , Resposta SOS em Genética , Salmonella typhimurium/fisiologia , Salmonella typhimurium/efeitos da radiação , Estresse Fisiológico , Perfilação da Expressão Gênica , Genes Reporter , Viabilidade MicrobianaRESUMO
The European REACH regulation requires the use of animal experimentation to assess the risk of industrial chemicals. However, the 3R principle (reduction, replacement, refinement) demands the use of suitable alternative test methods. Many dossiers submitted for the authorization of chemicals have attempted to provide the required data without performing new experiments, relying heavily on in silico methods; in vitro assays were scarcely used. We propose a methodology that uses physiologically based toxicokinetic (PBTK) models to extrapolate in vitro data to the in vivo level. We collected experimental results for in vitro and in vivo ethoxyresorufin-O-deethylase and vitellogenin induction following chemical exposure and compared those results with model predictions. We found that the predictive power of aqueous chemical concentrations was limited; median effect concentrations (EC50s) based on internal concentrations in fish correlated better with in vitro EC50s. Our data show that in vitro assays could offer a substitute for fish studies when combined with PBTK models.
Assuntos
Modelos Biológicos , Oncorhynchus mykiss/metabolismo , Poluentes Químicos da Água/farmacocinética , Animais , Simulação por Computador , Citocromo P-450 CYP1A1/biossíntese , Indução Enzimática/efeitos dos fármacos , Toxicocinética , Vitelogeninas/biossíntese , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
The release of endocrine-disrupting compounds (EDCs) to the environment poses a health hazard to both humans and wildlife. EDCs can activate or inhibit endogenous endocrine functions by binding hormone receptors, leading to potentially adverse effects. Conventional analytical methods can detect EDCs at a high sensitivity and precision, but are blind to the biological activity of the detected compounds. To overcome this limitation, yeast-based bioassays have previously been developed as a pre-screening method, providing an effect-based overview of hormonal-disruptive activity within the sample prior to the application of analytical methods. These yeast biosensors express human endocrine-specific receptors, co-transfected with the relevant response element fused to the specific fluorescent protein reporter gene. We describe several molecular manipulations of the sensor/reporter circuit in a Saccharomyces cerevisiae bioreporter strain that have yielded an enhanced detection of estrogenic-like compounds. Improved responses were displayed both in liquid culture (96-well plate format) as well as in conjunction with sample separation using high-performance thin-layer chromatography (HPTLC). The latter approach allows for an assessment of the biological effect of individual sample components without the need for their chemical identification at the screening stage.
Assuntos
Técnicas Biossensoriais , Estrogênios , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Humanos , Disruptores Endócrinos/análise , Engenharia GenéticaRESUMO
Aquatic animals and consumers of aquatic animals are exposed to increasingly complex mixtures of known and as-yet-unknown chemicals with dioxin-like toxicities in the water cycle. Effect- and cell-based bioanalysis can cover known and yet unknown dioxin and dioxin-like compounds as well as complex mixtures thereof but need to be standardized and integrated into international guidelines for environmental testing. In an international laboratory testing (ILT) following ISO/CD 24295 as standard procedure for rat cell-based DR CALUX un-spiked and spiked extracts of drinking-, surface-, and wastewater were validated to generate precision data for the development of the full ISO-standard. We found acceptable repeatability and reproducibility ranges below 36 % by DR CALUX bioassay for the tested un-spiked and spiked water of different origins. The presence of 17 PCDD/Fs and 12 dioxin-like PCBs was also confirmed by congener-specific GC-HRMS analysis. We compared the sum of dioxin-like activity levels measured by DR CALUX bioassay (expressed in 2,3,7,8-TCDD Bioanalytical Equivalents, BEQ; ISO 23196, 2022) with the obtained GC-HRMS chemical analysis results converted to toxic equivalents (TEQ; van den Berg et al., 2013).
Assuntos
Dioxinas , Bifenilos Policlorados , Dibenzodioxinas Policloradas , Ratos , Animais , Dibenzodioxinas Policloradas/análise , Dioxinas/toxicidade , Dioxinas/análise , Águas Residuárias , Reprodutibilidade dos Testes , Dibenzofuranos/análise , Rios , Luciferases , Bifenilos Policlorados/análise , Bioensaio/métodos , Dibenzofuranos Policlorados/análiseRESUMO
The present study investigated the hypothesis that the coupling of high-performance thin-layer chromatography with the yeast estrogen screen (planar-YES, p-YES) can be used as a screening tool for effect-directed analysis. Therefore, the proposed method was challenged for the first time with several real samples from various origins such as sediment pore water, wastewater, and sunscreens. It was possible to detect and quantify estrogenic effects in all investigated sample types, even in the presence of demanding matrixes. Furthermore, the specific agonistic effect of the estrogen receptor activation could be detected in samples exhibiting cytotoxic effects and at cytotoxic levels of analyzed estrogenic compounds, which is not possible with the classic YES. The analysis of samples by the p-YES results in profiles of estrogenic activity. By means of this profiles samples can be compared qualitatively and quantitatively with respect to different compositions of bioactive compounds in mixtures. In conclusion, the p-YES approach seems to have a high potential to be used as a valuable screening tool for various applications in effect-directed analysis.
Assuntos
Bioensaio/métodos , Cromatografia em Camada Fina/métodos , Estrogênios/análise , Estrogênios/isolamento & purificação , Sedimentos Geológicos/química , Protetores Solares/química , Águas Residuárias/químicaRESUMO
Many environmental mutagens, including polyaromatic compounds are present in surface waters, often in complex mixtures and at low concentrations. The present study provides and applies a novel, integrated approach to isolate polyaromatic mutagens in river water using a sample from the River Elbe. The sample was taken downstream of industrial discharges using blue rayon (BR) as a passive sampler that selectively adsorbs polyaromatic compounds and was subjected to effect-directed fractionation in order to characterise the compounds causing the detected effect(s). The procedure relies on three complementary fractionation steps, the Ames fluctuation assay with strains TA98, YG1024 and YG1041 with and without S9 activation and analytical screening. Several mutagenic fractions were isolated by combining mutagenicity testing with fractionation. The enhanced mutagenicity in the nitroreductase and/or O-acetyltransferase overexpressing strains YG1024 and YG1041 strains suggested amino- and/or nitro-compounds causing mutagenicity in several fractions. Analytical screening of mutagenic fractions with LC-HRMS/MS provided a list of molecular formulas typically containing one to ten nitrogen and at least two oxygen atoms supporting the presence of amino and nitro-compounds in the mutagenic fractions.
Assuntos
Técnicas Biossensoriais/métodos , Testes de Mutagenicidade/métodos , Mutagênicos/análise , Rios/química , Poluentes Químicos da Água/análise , Técnicas Biossensoriais/instrumentação , Mutagênicos/toxicidade , Salmonella typhimurium/efeitos dos fármacos , Poluentes Químicos da Água/toxicidadeRESUMO
Anti-androgens entering the aquatic environment, e.g., by effluents from wastewater treatment plants or agricultural settings are contributing to endocrine disruption in wildlife and humans. Due to the simultaneous presence of agonistic compounds, common in vitro bioassays can underestimate the risk posed by androgen antagonists. On the other hand, cytotoxic effects might lead to false positive assessments of anti-androgenic effects in conventional bioassays. In the present study, a combination of normal phase high-performance thin-layer chromatography (NP-HPTLC) with a yeast-based reporter gene assay is established for the detection of anti-androgenicity as a promising tool to reduce interferences of androgenic and anti-androgenic compounds present in the same sample. To avoid a misinterpretation of anti-androgenicity with cytotoxic effects, cell viability was assessed in parallel on the same plate using a resazurin viability assay adapted to HPTLC plates. The method was characterized by establishing dose-response curves for the model compounds flutamide and bisphenol A. Calculated effective doses at 10% (ED10) were 27.9 ± 1.3 ng zone-1 for flutamide and 20.1 ± 5.1 ng zone-1 for bisphenol A. Successful distinction between anti-androgenicity and cytotoxicity was exemplarily demonstrated with 4-nitroquinoline 1-oxide. As a proof of concept, the detection and quantification of anti-androgenicity in an extract of a landfill leachate is demonstrated. This study shows that the hyphenation of HPTLC with the yeast anti-androgen screen is a matrix-robust, cost-efficient and fast screening tool for the sensitive and simultaneous detection of anti-androgenic and cytotoxic effects in environmental samples. The method offers a wide range of possible applications in environmental monitoring and contributes to the identification of anti-androgenicity drivers in the course of an effect-directed analysis.
Assuntos
Antagonistas de Androgênios , Androgênios , Humanos , Androgênios/toxicidade , Antagonistas de Androgênios/toxicidade , Saccharomyces cerevisiae , Flutamida , Bioensaio/métodos , Cromatografia em Camada Fina/métodosRESUMO
Global ocean warming results in an increase of infectious diseases including an elevated emergence of Vibrio spp. in Northern Europe. The European Centre for Disease Prevention and Control reported annual periods of high to very high risks of infection with Vibrio spp. during summer months along the North Sea and Baltic Sea coasts. Based on those facts, the risk of Vibrio infections associated with recreational bathing in European coastal waters increases. To obtain an overview of the seasonal and spatial distribution of potentially human pathogenic Vibrio spp. at German coasts, this study monitored V. cholerae, V. parahaemolyticus, and V. vulnificus at seven recreational bathing areas from 2017 to 2018, including the heat wave event in summer 2018. The study shows that all three Vibrio species occurred in water and sediment samples at all sampling sites. Temperature was shown to be the main driving factor of Vibrio abundance, whereas Vibrio community composition was mainly modulated by salinity. A species-specific rapid increase was observed at water temperatures above 10°C, reaching the highest detection numbers during the heat wave event with abundances of 4.5 log10 CFU+1/100 ml of seawater and 6.5 log10 CFU+1/100 g of sediment. Due to salinity, the dominant Vibrio species found in North Sea samples was V. parahaemolyticus, whereas V. vulnificus was predominantly detected in Baltic Sea samples. Most detections of V. cholerae were associated with estuarine samples from both seas. Vibrio spp. concentrations in sediments were up to three log higher compared to water samples, indicating that sediments are an important habitat for Vibrio spp. to persist in the environment. Antibiotic resistances were found against beta-lactam antibiotics (ampicillin 31%, cefazolin 36%, and oxacillin and penicillin 100%) and trimethoprim-sulfamethoxazole (45%). Moreover, isolates harboring pathogenicity-associated genes such as trh for V. parahaemolyticus as well as vcg, cap/wcv, and the 16S rRNA-type B variant for V. vulnificus were detected. All sampled V. cholerae isolates were identified as non-toxigenic non-O1/non-O139 serotypes. To sum up, increasing water temperatures at German North Sea and Baltic Sea coasts provoke elevated Vibrio numbers and encourage human recreational water activities, resulting in increased exposure rates. Owing to a moderate Baltic Sea salinity, the risk of V. vulnificus infections is of particular concern.
Assuntos
Vibrioses , Vibrio cholerae , Vibrio parahaemolyticus , Vibrio , Humanos , Prevalência , RNA Ribossômico 16S , Vibrio/genética , Vibrioses/epidemiologia , Vibrio cholerae/genética , Vibrio parahaemolyticus/genética , ÁguaRESUMO
Sediment extracts from three polluted sites of the river Elbe basin were fractionated using a novel online fractionation procedure. Resulting fractions were screened for mutagenic, aryl hydrocarbon receptor (AhR)-mediated, transthyretin (TTR)-binding, and estrogenic activities and their potency to inhibit gap junctional intercellular communication (GJIC) to compare toxicity patterns and identify priority fractions. Additionally, more than 200 compounds and compound classes were identified using GC-MS/MS, LC-MS/MS, and HPLC-DAD methods. For all investigated end points, major activities were found in polar fractions, which are defined here as fractions containing dominantly compounds with at least one polar functional group. Nonpolar PAH fractions contributed to mutagenic and AhR-mediated activities while inhibition of GJIC and estrogenic and TTR-binding activities were exclusively observed in the polar fractions. Known mutagens in polar fractions included nitro- and dinitro-PAHs, azaarenes, and keto-PAHs, while parent and monomethylated PAHs such as benzo[a]pyrene and benzofluoranthenes were identified in nonpolar fractions. Additionally, for one sample, high AhR-mediated activities were determined in one fraction characterized by PCDD/Fs, PCBs, and PCNs. Estrone, 17ß-estradiol, 9H-benz[de]anthracen-7-one, and 4-nonylphenol were identified as possible estrogenic and TTR-binding compounds. Thus, not only nonpolar compounds such as PAHs, PCBs, and PCDD/Fs but also the less characterized and investigated more polar substances should be considered as potent mutagenic, estrogenic, AhR-inducing, TTR-binding, and GJIC-inhibiting components for future studies.
Assuntos
Sedimentos Geológicos/química , Poluentes Químicos da Água/análise , Animais , Bioensaio , Fracionamento Químico , Disruptores Endócrinos/análise , Disruptores Endócrinos/química , Disruptores Endócrinos/toxicidade , Monitoramento Ambiental , Alemanha , Humanos , Mutagênicos/análise , Mutagênicos/química , Mutagênicos/toxicidade , Pré-Albumina/análise , Pré-Albumina/química , Ratos , Receptores de Hidrocarboneto Arílico/análise , Receptores de Hidrocarboneto Arílico/química , Testes de Toxicidade , Poluentes Químicos da Água/química , Poluentes Químicos da Água/toxicidadeRESUMO
A bacterial genotoxicity reporter strain was constructed in which the tightly controlled strong promoter of the Escherichia coli SOS response gene sulA was fused to the alkaline phosphatase-coding phoA reporter gene. The bioreporter responded in a dose-dependent manner to three model DNA-damaging agents-hydrogen peroxide, nalidixic acid (NA), and mitomycin C (MMC)-detected 30-60 min after exposure. Detection thresholds were 0.15 µM for MMC, 7.5 µM for nalidixic acid, and approximately 50 µM for hydrogen peroxide. A similar response to NA was observed when the bioreporter was integrated into a specially designed, portable electrochemical detection platform. Reporter sensitivity was further enhanced by single and double knockout mutations that enhanced cell membrane permeability (rfaE) and inhibited DNA damage repair mechanisms (umuD, uvrA). The rfaE mutants displayed a five- and tenfold increase in sensitivity to MMC and NA, respectively, while the uvrA mutation was advantageous in the detection of hydrogen peroxide. A similar sensitivity was displayed by the double rfaE/uvrA mutant when challenged with the pre-genotoxic agents 2-amino-3-methylimidazo[4,5-f]quinoline and 2-aminoanthracene following metabolic activation with an S9 mammalian liver fraction.
Assuntos
Técnicas Biossensoriais/métodos , Proteínas de Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Técnicas Eletroquímicas/métodos , Proteínas de Escherichia coli/metabolismo , Genes Reporter , Peróxido de Hidrogênio/toxicidade , Mitomicina/toxicidade , Ácido Nalidíxico/toxicidade , Regiões Promotoras Genéticas , Resposta SOS em GenéticaRESUMO
Tyre and road wear is one of the main emission sources of particulate plastics (microplastics). In this study, the emissions of tyre wear particles (TWP) which are annually generated on the German road network were calculated. Emissions are calculated by applying two different data sets of emission factors and the annual mileage for distinct vehicles and road types (urban roads, rural roads, highways). Environmental entry paths of tyre and road wear particles (TRWP) were considered including releases to ambient air, soils and surface waters. Road runoff treatment was taken into account differentiated into sewage systems in urban areas and at non-urban roads. This study identifies the soundest data available concerning emission factors, data on traffic and road systems as well as comprehensive information on road runoff treatment in Germany. Applying this data and assuming that 5% of the total emissions are 'fine' air-borne particles, the emissions of coarse 'non-airborne' particles are 75,200-98,400 t/a; . the transport to road banks and soils near roads is 57,300-65,400 t/a (66-76%) including runoff and drift; 8700-19,800 t/a (12-20%) are released to surface waters. Due to lack of data, degradation in soils and surface waters was not considered. Besides soils, urban wastewater treatment plants are expected to be an important sink of TRWP assuming that most of the particles are incorporated in the sewage sludge. Due to the application of sewage sludge as a fertilizer, 1400-2800 t/a TWP are currently deposited on agricultural areas. No reliable data was available to estimate the masses of TWP which are transported from freshwaters into the marine environment. Existing mitigation measures should be improved according to the principle of precaution by installation of road runoff treatment systems and retrofitting of existing plants regarding optimum fine particle retention.
RESUMO
The ubiquity of microplastics in aquatic ecosystems has raised concerns over their interaction with biota. However, microplastics research on freshwater species, especially mollusks, is still scarce. We, therefore, investigated the factors affecting microplastics ingestion in the freshwater mussel Dreissena polymorpha. Using polystyrene spheres (5, 10, 45, 90 µm), we determined the body burden of microplastics in the mussels in relation to 1) exposure and depuration time, 2) body size, 3) food abundance, and 4) microplastic concentrations. D. polymorpha rapidly ingested microplastics and excreted most particles within 12 h. A few microplastics were retained for up to 1 wk. Smaller individuals had a higher relative body burden of microplastics than larger individuals. The uptake of microplastics was concentration-dependent, whereas an additional food supply (algae) reduced it. We also compared the ingestion of microplastics by D. polymorpha with 2 other freshwater species (Anodonta anatina, Sinanodonta woodiana), highlighting that absolute and relative uptake depends on the species and the size of the mussels. In addition, we determined toxicity of polystyrene fragments (≤63 µm, 6.4-100 000 p mL-1 ) and diatomite (natural particle, 100 000 p mL-1 ) in D. polymorpha after 1, 3, 7, and 42 d of exposure, investigating clearance rate, energy reserves, and oxidative stress. Despite ingesting large quantities, exposure to polystyrene fragments only affected the clearance rate of D. polymorpha. Further, results of the microplastic and diatomite exposure did not differ significantly. Therefore, D. polymorpha is unaffected by or can compensate for polystyrene fragment toxicity even at concentrations above current environmental levels. Environ Toxicol Chem 2021;40:2247-2260. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
Assuntos
Bivalves , Poluentes Químicos da Água , Animais , Ingestão de Alimentos , Ecossistema , Monitoramento Ambiental , Água Doce , Humanos , Microplásticos/toxicidade , Plásticos/toxicidade , Poliestirenos/toxicidade , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
Organic coatings can guarantee long-term protection of steel structures due to causing a physical barrier against water and oxygen. Because of their mechanical properties and resistances to heat and chemicals, epoxy resin-based coatings are widely used for corrosion protection. Despite of the aromatic backbone and the resulting susceptibility to UV degradation, epoxy resins are frequently used as binding agent in top layers of anti-corrosion coating systems. Consequently, these organic polymers are directly exposed to sunlight and thus UV radiation. The present study was designed to investigate if toxic effects of epoxy resin-based-coatings are changed by UV-A irradiation. For this purpose, two epoxide-based top coatings were examined with and without UV aging for their bacterial toxicity and estrogenicity. In addition, chemical analyses were performed to identify released compounds as well as photolytic degradation products and to assign toxic effects to individual substances. UV-A irradiation of epoxy resin based top coatings resulted in an overall decrease of acute and specific ecotoxicological effects but as well to the formation of toxic transformation products. Both, in leachates of untreated and UV-A irradiated coatings, 4tBP was identified as the main driver of estrogenicity and toxicity to luminescent bacteria. BPA and structural analogs contributing to estrogenic effects in leachates were formed by UV-A irradiation. The combination of HPTLC coupled bioassays and LC-MS analyses supported the identification of bioactive compounds in terms of an effect-directed analysis. The present findings indicate that epoxide-based coatings are less suitable for the application as top coatings and more UV stable coatings like aliphatic polyurethanes should be preferred.
RESUMO
Microplastic abundances have been studied intensively in the last years in marine and freshwater environments worldwide. Though several articles have been published about the Mediterranean Sea, only few studies about the Black Sea exist. The Black Sea drains into the Mediterranean Sea and may therefore significantly contribute to the Mediterranean marine pollution. So far, only very few articles have been published about micro-, meso- and macroplastic abundances in the Western Black Sea. In order to fill this knowledge gap and to decipher the number of plastics on the water surface, 12 samples were collected from surface waters with a neustonic net (mesh size 200 µm) in the Black Sea close to the Danube Delta and the Romanian shore. Organic matter was digested and plastic particles were isolated by density separation. The results of visual inspection, pyrolysis GC-MS (for microplastics) and ATR-FTIR (for mesoplastics >5 mm) revealed an average concentration of 7 plastic particles/m³, dominated by fibers (â¼76%), followed by foils (â¼13%) and fragments (â¼11%). Only very few spherules were detected. The polymers polypropylene (PP) and polyethylene (PE) dominated which is in line with other studies analyzing surface waters from rivers in Western Europe as well as in China. Statistical analyses show that the plastic concentration close to the mouth of the Danube River was significantly higher than at four nearshore regions along the Romanian and Bulgarian coastline. This could be explained by plastic inputs from the Danube River into the western part of the Black Sea.
Assuntos
Plásticos , Poluentes Químicos da Água , Mar Negro , Bulgária , China , Monitoramento Ambiental , Europa (Continente) , Mar Mediterrâneo , Poluentes Químicos da Água/análiseRESUMO
Flooding and other sediment disturbances can lead to increases in sediment resuspension. In this context, it is of central importance to understand the kinetics of release from these sediments and the uptake of pollutants, such as polychlorinated biphenyls (PCBs) and polychlorinated dioxins and furans (PCDD/Fs), into aquatic organisms. In the present study, we parameterized a sediment desorption model based on experimentally determined rapidly-desorbing fractions of dioxin-like chemicals (DLCs). We coupled this desorption model with a physiologically-based toxicokinetic model for rainbow trout. This combined model was used to predict DLC concentrations in the muscle of exposed fish. The performance of this model was evaluated using a previously published dataset on DLC uptake from sediment suspensions during simulated re-suspension events. Predictions generally differed less than 10-fold from measured values, and the model showed a good global coefficient of determination (R2) of 0.95. The root mean squared error (RMSE) for PCBs was 0.31 log units and 0.53 log units for PCDD/Fs. The results of our study demonstrate that the prediction of bioconcentration and related risk to fish resulting from sediment resuspension can be accurately predicted using coupled desorption and toxicokinetic models.