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  • 1.
    Alfonso, Sébastien
    et al.
    Ifremer, Laboratoire Ressources Halieutiques, Place Gaby Coll, F-17137, L'Houmeau, France; UMR MARBEC, Ifremer, IRD, UM2, CNRS, Laboratoire Adaptation et Adaptabilités des Animaux et des Systèmes, Route de Maguelone, F-34250, Palavas-les-Flots, France.
    Blanc, Mélanie
    Örebro University, School of Science and Technology. Ifremer, Laboratoire Ressources Halieutiques, Place Gaby Coll, F-17137, L'Houmeau, France.
    Joassard, Lucette
    Ifremer, Laboratoire Ressources Halieutiques, Place Gaby Coll, F-17137, L'Houmeau, France.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Munschy, Catherine
    Ifremer, Laboratoire Biogéochimie des Contaminants Organiques, Rue de l'Ile d'Yeu, BP 21105, F-44311, Nantes, Cedex 3, France.
    Loizeau, Véronique
    Ifremer, Laboratoire Biogéochimie des Contaminants Organiques, ZI Pointe du Diable, CS 10070, F-29280, Plouzané, France.
    Bégout, Marie-Laure
    Ifremer, Laboratoire Ressources Halieutiques, Place Gaby Coll, F-17137, L'Houmeau, France.
    Cousin, Xavier
    UMR MARBEC, Ifremer, IRD, UM2, CNRS, Laboratoire Adaptation et Adaptabilités des Animaux et des Systèmes, Route de Maguelone, F-34250, Palavas-les-Flots, France; Inra, UMR GABI, Inra, AgroParisTech, Domaine de Vilvert, Batiment 231, F-78350 Jouy-en-Josas, France.
    Examining multi- and transgenerational behavioral and molecular alterations resulting from parental exposure to an environmental PCB and PBDE mixture2019In: Aquatic Toxicology, ISSN 0166-445X, E-ISSN 1879-1514, Vol. 208, p. 29-38Article in journal (Refereed)
    Abstract [en]

    Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants extensively used during the 20th century and still present in aquatic environments despite their ban. Effects of exposure to these compounds over generations are poorly documented. Therefore, our aims were to characterize behavioral responses and underlying molecular mechanisms in zebrafish exposed to an environmentally relevant mixture of PCBs and PBDEs as well as in four unexposed offspring generations. Zebrafish (F0) were chronically exposed from the first meal onward to a diet spiked with a mixture containing 22 PCB and 7 PBDE congeners in proportions and concentrations reflecting environmental situations (ΣPCBs = 1991 and ΣPBDEs = 411 ng/g). Four offspring generations (F1 to F4) were obtained from this F0 and were not further exposed. Behavior was assessed at both larval and adult stages. Mechanisms related to behavioral defects (habenula maturation and c-fos transcription) and methylation (dnmts transcription) were monitored in larvae. Exposed adult F0 as well as F1 and F3 adults displayed no behavioral change while F2 expressed anxiety-like behavior. Larval behavior was also disrupted, i.e. hyperactive after light to dark transition in F1 or hypoactive in F2, F3 and F4. Behavioral disruptions may be related to defect in habenula maturation (observed in F1) and change in c-fos transcription (observed in F1 and F2). Transcription of the gene encoding DNA methyltransferase (dnmt3ba) was also modified in all generations. Our results lead us to hypothesize that chronic dietary exposure to an environmentally relevant mixture of PCB and PBDE triggers multigenerational and transgenerational molecular and behavioral disruptions in a vertebrate model.

  • 2.
    Beiras, R.
    et al.
    ECIMAT, University of Vigo, Vigo, Galicia, Spain.
    Bellas, J.
    Centro Oceanográfico de Vigo, Instituto Español de Oceanografía, Vigo, Galicia, Spain.
    Cachot, J.
    Bordeaux University, EPOC, Talence, France.
    Cormier, Bettie
    Örebro University, School of Science and Technology.
    Cousin, X.
    IFREMER, Laboratoire Adaptation et Adaptabilités des Animaux et des Systèmes, UMR MARBEC, Palavas, France; UMR GABI INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, France.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Gambardella, C.
    CNR-ISMAR, Genova, Italy.
    Garaventa, F.
    CNR-ISMAR, Genova, Italy.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Le Bihanic, F.
    Bordeaux University, EPOC, Talence, France.
    López-Ibáñez, S.
    ECIMAT, University of Vigo, Vigo, Galicia, Spain.
    Piazza, V.
    CNR-ISMAR, Genova, Italy.
    Rial, D.
    Centro Oceanográfico de Vigo, Instituto Español de Oceanografía, Vigo, Galicia, Spain.
    Tato, T.
    ECIMAT, University of Vigo, Vigo, Galicia, Spain.
    Vidal-Liñán, L.
    Centro Oceanográfico de Vigo, Instituto Español de Oceanografía, Vigo, Galicia, Spain.
    Ingestion and contact with polyethylene microplastics does not cause acute toxicity on marine zooplankton2018In: Journal of Hazardous Materials, ISSN 0304-3894, E-ISSN 1873-3336, Vol. 360, p. 452-460Article in journal (Refereed)
    Abstract [en]

    Toxicity of polyethylene microplastics (PE-MP) of size ranges similar to their natural food to zooplanktonic organisms representative of the main taxa present in marine plankton, including rotifers, copepods, bivalves, echinoderms and fish, was evaluated. Early life stages (ELS) were prioritized as testing models in order to maximize sensitivity. Treatments included particles spiked with benzophenone-3 (BP-3), a hydrophobic organic chemical used in cosmetics with direct input in coastal areas. Despite documented ingestion of both virgin and BP-3 spiked microplastics no acute toxicity was found at loads orders of magnitude above environmentally relevant concentrations on any of the invertebrate models. In fish tests some effects, including premature or reduced hatching, were observed after 12 d exposure at 10 mg L-1 of BP-3 spiked PE-MP. The results obtained do not support environmentally relevant risk of microplastics on marine zooplankton. Similar approaches testing more hydrophobic chemicals with higher acute toxicity are needed before these conclusions could be extended to other organic pollutants common in marine ecosystems. Therefore, the replacement of these polymers in consumer products must be carefully considered.

  • 3.
    Blanc, Mélanie
    et al.
    Örebro University, School of Science and Technology.
    Kärrman, Anna
    Örebro University, School of Science and Technology.
    Kukučka, Petr
    Örebro University, School of Science and Technology.
    Scherbak, Nikolai
    Örebro University, School of Science and Technology.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Mixture-specific gene expression in zebrafish (Danio rerio) embryos exposed to perfluorooctane sulfonic acid (PFOS), perfluorohexanoic acid (PFHxA) and 3,3′,4,4′,5-pentachlorobiphenyl (PCB126)2017In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 590-591, p. 249-257Article in journal (Refereed)
    Abstract [en]

    Perfluorooctane sulfonic acid (PFOS) and 3,3′,4,4′,5-pentachlorobiphenyl (PCB126) are persistent organic pollutants of high concern because of their environmental persistence, bioaccumulation and toxic properties. Besides, the amphiphilic properties of fluorinated compounds such as PFOS and perfluorohexanoic acid (PFHxA) suggest a role in increasing cell membrane permeability and solubilizing chemicals. The present study aimed at investigating whether PFOS and PFHxA are capable of modifying the activation of PCB126 toxicity-related pathways. For this purpose, zebrafish embryos were exposed in semi-static conditions to 7.5 μg/L of PCB126 alone, in the presence of 25 mg/L of PFOS, 15.7 mg/L of PFHxA or in the presence of both PFOS and PFHxA. Quantitative PCR was performed on embryos aged from 24 h post fertilization (hpf) to 96 hpf to investigate expression changes of genes involved in metabolism of xenobiotics (ahr2, cyp1a), oxidative stress (gpx1a, tp53), lipids metabolism (acaa2, osbpl1a), and epigenetic mechanisms (dnmt1, dnmt3ba). Cyp1a and ahr2 expression were significantly induced by the presence of PCB126. However, after 72 and 78 h of exposure, induction of cyp1a expression was significantly lower when embryos were co-exposed to PCB126 + PFOS + PFHxA when compared to PCB126-exposed embryos. Significant upregulation of gpx1a occurred after exposure to PCB126 + PFHxA and to PCB126 + PFOS + PFHxA at 30 and 48 hpf. Besides, embryos appeared more sensitive to PCB126 + PFOS + PFHxA at 78 hpf: acaa2 and osbpl1a were significantly downregulated; dnmt1 was significantly upregulated. While presented as environmentally safe, PFHxA demonstrated that it could affect gene expression patterns in zebrafish embryos when combined to PFOS and PCB126, suggesting that such mixture may increase PCB126 toxicity. This is of particular relevance since PFHxA is persistent and still being ejected into the environment. Moreover, it provides additional information as to the importance to integrate mixture effects of chemicals in risk assessment and biomonitoring frameworks.

  • 4.
    Blanc, Mélanie
    et al.
    Örebro University, School of Science and Technology.
    Rüegg, Joëlle
    Institute for Environmental Medicine, Karolinska Institutet, Solna, Sweden.
    Scherbak, Nikolai
    Örebro University, School of Science and Technology.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Environmental chemicals differentially affect epigenetic-related mechanisms in the zebrafish liver (ZF-L) cell line and in zebrafish embryos2019In: Aquatic Toxicology, ISSN 0166-445X, E-ISSN 1879-1514, Vol. 215, article id 105272Article in journal (Refereed)
    Abstract [en]

    A number of chemicals have been shown to affect epigenetic patterning and functions. Since epigenetic mechanisms regulate transcriptional networks, epigenetic changes induced by chemical exposure can represent early molecular events for long-term adverse physiological effects. Epigenetics has thus appeared as a research field of major interest within (eco)toxicological sciences. The present study aimed at measuring effects on epigenetic-related mechanisms of selected environmental chemicals (bisphenols, perfluorinated chemicals, methoxychlor, permethrin, vinclozolin and coumarin 47) in zebrafish embryos and liver cells (ZFL). Transcription of genes related to DNA methylation and histone modifications was measured and global DNA methylation was assessed in ZFL cells using the LUMA assay. The differences in results gathered from both models suggest that chemicals affect different mechanisms related to epigenetics in embryos and cells. In zebrafish embryos, exposure to bisphenol A, coumarin 47, methoxychlor and permethrin lead to significant transcriptional changes in epigenetic factors suggesting that they can impact early epigenome reprogramming related to embryonic development. In ZFL cells, significant transcriptional changes were observed upon exposure to all chemicals but coumarin 47; however, only perfluorooctane sulfonate induced significant effects on global DNA methylation. Notably, in contrast to the other tested chemicals, perfluorooctane sulfonate affected only the expression of the histone demethylase kdm5ba. In addition, kdm5ba appeared as a sensitive gene in zebrafish embryos as well. Taken together, the present results suggest a role for kdm5ba in regulating epigenetic patterns in response to chemical exposure, even though mechanisms remain unclear. To confirm these findings, further evidence is required regarding changes in site-specific histone marks and DNA methylation together with their long-term effects on physiological outcomes.

  • 5.
    Bluhm, Kerstin
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research, Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Otte, Jens
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
    Yang, Lee
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
    Zinsmeister, Christian
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
    Legradi, Jessica
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany; Institute for Environmental Studies, VU University Amsterdam, Amsterdam, The Netherlands.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research, Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Kosmehl, Thomas
    Aquatic Ecology and Toxicology Group, Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Group, Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Strähle, Uwe
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany; School of Environment, Nanjing University, Nanjing, China; Key Laboratory of Yangtze River Environment of Education Ministry of China, College of Environmental Science and Engineering, Tongji University, Shanghai, China; College of Resources and Environmental Science, Chongqing University, Chongqing, China.
    Impacts of Different Exposure Scenarios on Transcript Abundances in Danio rerio Embryos when Investigating the Toxicological Burden of Riverine Sediments2014In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 9, no 9, article id e106523Article in journal (Refereed)
    Abstract [en]

    Purpose: Recently, a proof-of-concept study revealed the suitability of transcriptome analyses to obtain and assess changes in the abundance of transcripts in zebrafish (Danio rerio) embryos after exposure to organic sediment extracts. The present study investigated changes in the transcript abundance in zebrafish embryos exposed to whole sediment samples and corresponding organic extracts in order to identify the impact of different exposure pathways on sediment toxicity.

    Materials and Methods: Danio rerio embryos were exposed to sublethal concentrations of three sediment samples from the Danube River, Germany. The sediment samples were investigated both as freeze-dried samples and as organic extracts. Silica dust and a process control of the extraction procedure were used as references. After exposure, mRNA was isolated and changes in profiles of gene expression levels were examined by an oligonucleotide microarray. The microarray results were compared with bioassays, chemical analysis of the sediments and profiles of gene expression levels induced by several single substances.

    Results and Discussion: The microarray approach elucidated significant changes in the abundance of transcripts in exposed zebrafish embryos compared to the references. Generally, results could be related to Ah-receptor-mediated effects asconfirmed by bioassays and chemical analysis of dioxin-like contaminants, as well as to exposure to stress-inducing compounds. Furthermore, the results indicated that mixtures of chemicals, as present in sediment and extract samples, result in complex changes of gene expression level profiles difficult to compare with profiles induced by single chemical substances. Specifically, patterns of transcript abundances were less influenced by the chemical composition at the sampling site compared t the method of exposure (sediment/extract). This effect might be related to different bioavailability of chemicals.

    Conclusions: The apparent difference between the exposure scenarios is an important aspect that needs to be addressed when conducting analyses of alterations in the expression level of mRNA.

  • 6.
    Bour, Agathe
    et al.
    Department of Biosciences, University of Oslo, Oslo, Norway.
    Haarr, Ane
    Department of Biosciences, University of Oslo, Oslo, Norway.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Hylland, Ketil
    Department of Biosciences, University of Oslo, Oslo, Norway.
    Environmentally relevant microplastic exposure affects sediment-dwelling bivalves2018In: Environmental Pollution, ISSN 0269-7491, E-ISSN 1873-6424, Vol. 236, p. 652-660Article in journal (Refereed)
    Abstract [en]

    Most microplastics are expected to sink and end up in marine sediments. However, very little is known concerning their potential impact on sediment-dwelling organisms. We studied the long-term impact of microplastic exposure on two sediment-dwelling bivalve species. Ennucula tenuis and Abra nitida were exposed to polyethylene microparticles at three concentrations (1; 10 and 25 mg/kg of sediment) for four weeks. Three size classes (4-6; 20-25 and 125-500 mu m) were used to study the influence of size on microplastic ecotoxicity. Microplastic exposure did not affect survival, condition index or burrowing behaviour in either bivalve species. However, significant changes in energy reserves were observed. No changes were observed in protein, carbohydrate or lipid contents in E. tenuis, with the exception of a decrease in lipid content for one condition. However, total energy decreased in a dose-dependent manner for bivalves exposed to the largest particles. To the contrary, no significant changes in total energy were observed for A. nitida, although a significant decrease of protein content was observed for individuals exposed to the largest particles, at all concentrations. Concentration and particle size significantly influenced microplastic impacts on bivalves, the largest particles and higher concentrations leading to more severe effects. Several hypotheses are presented to explain the observed modulation of energy reserves, including the influence of microplastic size and concentration. Our results suggest that long-term exposure to microplastics at environmentally relevant concentrations can impact marine benthic biota.

  • 7.
    Bräunig, Jennifer
    et al.
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany; National Research Centre for Environmental Toxicology (Entox), The University of Queensland, Brisbane, Australia.
    Schiwy, Sabrina
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Broedel, Oliver
    Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Wildau, Germany.
    Müller, Yvonne
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Frohme, Marcus
    Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Wildau, Germany.
    Hollert, Henner
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Time-dependent expression and activity of cytochrome P450 1s in early life-stages of the zebrafish (Danio rerio)2015In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 22, no 21, p. 16319-15328Article in journal (Refereed)
    Abstract [en]

    Zebrafish embryos are being increasingly used as model organisms for the assessment of single substances and complex environmental samples for regulatory purposes.Thus, it is essential to fully understand the xenobiotic metabolism during the different life-stages of early development.The aim of the present study was to determine arylhydrocarbon receptor (AhR)-mediated activity during selected times of early development using qPCR, enzymatic activity through measurement of 7-ethoxyresorufin-Odeethylase(EROD) activity, and protein expression analysis. In the present study, gene expression of cyp1a, cyp1b1, cyp1c1, cyp1c2, and ahr2 as well as EROD activity were investigated up to 120 h postfertilization (hpf) after exposure to either β-naphthoflavone (BNF) or a polycyclic aromatichydrocarbons (PAH)-contaminated sediment extract from Vering Kanal in Hamburg (VK). Protein expression was measured at 72 hpf after exposure to 20 μg/L BNF. Altered proteins were identified by matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) peptide mass fingerprinting. Distinct patterns of basal messenger RNA (mRNA) expressionwere found for each of the cyp1 genes, suggesting specific roles during embryonic development. All transcripts were induced by BNF and VK. ahr2 mRNA expression was significantly upregulated after exposure toVK. All cyp1 genes investigated showed a temporal decline in expression at 72 hpf. The significant decline of Hsp 90β protein at 72 hpf after exposure to BNF may suggest an explanation for the decline of cyp1 genes at this time point as Hsp 90β is of major importance for the functioning of the Ah-receptor. EROD activity measured in embryos was significantly induced after 96 hpf of exposure to BNF or VK. Together, these results demonstrate distinct temporal patterns of cyp1 genes and protein activities in zebrafish embryos as well as show a need to investigate further the xenobiotic biotransformation system during early development of zebrafish.

  • 8.
    Böttcher, Melanie
    et al.
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Grund, Stefanie
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Keiter, Steffen
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Kosmehl, Thomas
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Reifferscheid, Georg
    German Federal Institute of Hydrology, Koblenz, Germany.
    Seitz, Nadja
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Suares Rocha, Paula
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Hollert, Henner
    Institute for Environmental Research (Biology V), Department for Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Comparison of in vitro and in situ genotoxicity in the Danube River by means of the comet assay and the micronucleus test2010In: Mutation research. Genetic toxicology and environmental mutagenesis, ISSN 1383-5718, E-ISSN 1879-3592, Vol. 700, no 1-2, p. 11-17Article in journal (Refereed)
    Abstract [en]

    Genotoxicity can be correlated with adverse reproductive effects or may even result in elevated extinction risk for particular species of an ecosystem. It may thus be a valuable tool for screening of pollution and potential environmental harm. Since many genotoxicants tend to adsorb on to particulate matter, sediments and suspended materials are of particular interest for genotoxicity screening under field conditions. In order to correlate the genotoxic potential of sediments with genetic damage in fish, rainbow-trout liver (RTL-W1) cells were exposed in vitro to acetone extracts of sediments collected at 10 selected sites along the upper Danube River and analyzed in the comet and micronucleus assays. These in vitro results were compared with micronucleus formation in erythrocytes of the European barbel (Barbus barbus) caught int he field. The two in vitro bioassays showed excellent correlation, indicating comparability of genotoxic potentials in vitro. Sampling sites could be clearly differentiated with respect to severity of effects, with Rottenacker as the most heavily contaminated site, Ehingen and Schwarzach as moderately genotoxic,and with the weakest effects in the tributary Lauchert. All other sediment extracts showed intermediate genotoxic or clastogenic effects. In situ, micronucleus formation in barbel erythrocytes indicated severe genotoxicity at Rottenacker, moderate effects at Ehingen, but minor contamination at Riedlingen and Sigmaringen. In situ observations thus showed excellent correlation with corresponding in vitro tests and document the ecological relevance of in vitro studies with sediment extracts. With respect to the ecological status of the Danube River, the results overall indicate a moderate to severe genotoxic potential with a highly differential localization.

  • 9.
    Cormier, Bettie
    et al.
    Örebro University, School of Science and Technology. University of Bordeaux, EPOC UMR CNRS 5805, Pessac, France.
    Batel, Annika
    Aquatic Ecology and Toxicology Group, Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Cachot, Jerome
    University of Bordeaux, EPOC UMR CNRS 5805, Pessac, France.
    Begout, Marie-Laure
    Laboratoire Ressources Halieutiques, IFREMER, L’Houmeau, France.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Group, Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Cousin, Xavier
    IFREMER, L3AS, UMR MARBEC, Palavas-les-Flots, France; UMR GABI INRA, AgroParisTech, University Paris-Saclay, Jouy-en-Josas, France.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Multi-Laboratory Hazard Assessment of Contaminated Microplastic Particles by Means of Enhanced Fish Embryo Test With the Zebrafish (Danio rerio)2019In: Frontiers in Environmental Science, E-ISSN 2296-665X, Vol. 7, article id 135Article in journal (Refereed)
    Abstract [en]

    As wide-spread pollutants in the marine environment, microplastics (MPs) have raised public concern about potential toxic effects in aquatic organisms, and, among others, MPs were suspected to act as a vector for organic pollutants to biota. The purpose of the present study was to investigate effects by three model pollutants, oxybenzone (BP3), benzo[a] pyrene (BaP), and perfluorooctane sulfonate (PFOS) adsorbed to polyethylene MPs on the basis of a standard assay, the acute fish embryo toxicity test (FET; OECD TG 236) with zebrafish (Danio rerio) supplemented by additional endpoints such as induction of ethoxyresorufin-O-deethylase (EROD) activity, modification of cyp1a gene transcription and changes in larval swimming behavior. FET assays were performed in three laboratories using slightly different husbandry and exposure conditions, which, however, were all fully compatible with the limits defined by OECD TG 236. This allowed for testing of potential changes in the FET assay due to protocol variations. The standard endpoints of the FET (acute embryotoxicity) did not reveal any acute toxicity for both virgin MPs and MPs spiked with BP3, BaP, and PFOS. With respect to sublethal endpoints, EROD activity was increased after exposure to MPs spiked with BP3 (3 h pulse) and MPs spiked with BaP (96 h continuous exposure). Cyp1a transcription was increased upon exposure to MPs spiked with BP3 or BaP. For the selected combination of MPs particles and contaminants, the basic FET proved not sensitive enough to reveal effects of (virgin and spiked) MPs. However, given that the FET can easily be supplemented by a broad variety of more subtle and sensitive endpoints, an enhanced FET protocol may provide a relevant approach with developmental stages of a vertebrate animal model, which is not protected by current EU animal welfare legislation (Directive EU 2010/63).

  • 10.
    Di Paolo, Carolina
    et al.
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Ottermanns, Richard
    Institute for Environmental Research, RWTH Aachen University, Aachen, GermanyWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Ait-Aissa, Selim
    INERIS, Verneuil-en-Halatte, France.
    Bluhm, Kerstin
    Institute for Environmental Research, Rheinisch-Westfälische Technische Hochschule, Aachen, Germany.
    Brack, Werner
    UFZ-Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Breitholtz, Magnus
    Department of Applied Environmental Science - ITM, Stockholm University, Stockholm, Sweden.
    Buchinger, Sebastian
    Department Biochemistry and Ecotoxicology, Federal Institute of Hydrology, Koblenz, Germany.
    Carere, Mario
    Italian Institute of Health, Rome, Italy.
    Chalon, Carole
    ISSeP (Scienti fi c Institute of Public Service), Liège, Belgium.
    Cousin, Xavier
    Laboratoire d'Ecotoxicologie, L'Institut Français de Recherche pour l'Exploitation de la Mer (Ifremer), L'Houmeau, France; Laboratoire de Physiologie et Génétique des Poissons, Inra, Rennes, France.
    Dulio, Valeria
    Institut national de l'environnement industriel et des risques (INERIS), Verneuil-en-Halatte, France.
    Escher, Beate
    UFZ-Helmholtz Centre for Environmental Research, Leipzig, Germany; National Research Centre for Environmental Toxicology - Entox, The University of Queensland, Brisbane, Australia; Centre for Applied Geosciences, Eberhard Karls University, Tübingen, Germany.
    Hamers, Timo
    Institute for Environmental Studies -IVM, Vrije University, Amsterdam, The Netherlands.
    Hilscherová, Klara
    Research Centre for Toxic Compounds in the Environment - RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic.
    Jarque, Sergio
    Research Centre for Toxic Compounds in the Environment - RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic.
    Jonas, Adam
    Research Centre for Toxic Compounds in the Environment - RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic.
    Maillot-Marechal, Emmanuelle
    Institut national de l'environnement industriel et des risques (INERIS), Verneuil-en-Halatte, France.
    Marneffe, Yves
    ISSeP (Scienti fi c Institute of Public Service), Liège, Wallonia, Belgium.
    Nguyen, Mai Thao
    Waterproef Laboratory, Edam, The Netherlands.
    Pandard, Pascal
    INERIS, Verneuil-en-Halatte, France.
    Schifferli, Andrea
    Swiss Centre for Applied Ecotoxicology Eawag-EPFL, Dübendorf, Switzerland.
    Schulze, Tobias
    UFZ-Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Seidensticker, Sven
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany; Laboratoire d'Ecotoxicologie, Ifremer, L'Houmeau, France.
    Seiler, Thomas B.
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Tang, Janet
    National Research Centre for Environmental Toxicology - Entox, The University of Queensland, Brisbane, Australia.
    van der Oost, Ron
    WATERNET Institute for the Urban Water Cycle, Division of Technology Research & Engineering, Amsterdam, The Netherlands.
    Vermeirssen, Etienne
    Swiss Centre for Applied Ecotoxicology Eawag-EPFL, Dübendorf, Switzerland.
    Zounková, Radka
    Research Centre for Toxic Compounds in the Environment - RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic.
    Zwart, Nick
    Institute for Environmental Studies -IVM, Vrije University, Amsterdam, The Netherlands.
    Hollert, Henner
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Bioassay battery interlaboratory investigation of emerging contaminants in spikedwater extracts: Towards the implementation of bioanalytical monitoring tools inwater quality assessment and monitoring2016In: Water Research, ISSN 0043-1354, Vol. 104, p. 473-484Article in journal (Refereed)
    Abstract [en]

    Bioassays are particularly useful tools to link the chemical and ecological assessments in water quality monitoring. Different methods cover a broad range of toxicity mechanisms in diverse organisms, and account for risks posed by non-target compounds and mixtures. Many tests are already applied in chemical and waste assessments, and stakeholders from the science-police interface have recommended their integration in regulatory water quality monitoring. Still, there is a need to address bioassay suitability to evaluate water samples containing emerging pollutants, which are a current priority in water quality monitoring. The presented interlaboratory study (ILS) verified whether a battery of miniaturized bioassays, conducted in 11 different laboratories following their own protocols, would produce comparable results when applied to evaluate blinded samples consisting of a pristine water extract spiked with four emerging pollutants as single chemicals or mixtures, i.e. triclosan, acridine, 17α-ethinylestradiol (EE2) and 3-nitrobenzanthrone (3-NBA). Assays evaluated effects on aquatic organisms from three different trophic levels (algae, daphnids, zebrafish embryos) and mechanism-specific effects using in vitro estrogenicity (ER-Luc, YES) and mutagenicity (Ames fluctuation) assays. The test battery presented complementary sensitivity and specificity to evaluate the different blinded water extract spikes. Aquatic organisms differed in terms of sensitivity to triclosan (algae > daphnids > fish) and acridine (fish > daphnids > algae) spikes, confirming the complementary role of the three taxa for water quality assessment. Estrogenicity and mutagenicity assays identified with high precision the respective mechanism-specific effects of spikes even when non-specific toxicity occurred in mixture. For estrogenicity, although differences were observed between assays and models, EE2 spike relative induction EC50 values were comparable to the literature, and E2/EE2 equivalency factors reliably reflected the sample content. In the Ames, strong revertant induction occurred following 3-NBA spike incubation with the TA98 strain, which was of lower magnitude after metabolic transformation and when compared to TA100. Differences in experimental protocols, model organisms, and data analysis can be sources of variation, indicating that respective harmonized standard procedures should be followed when implementing bioassays in water monitoring. Together with other ongoing activities for the validation of a basic bioassay battery, the present study is an important step towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.

  • 11.
    Di Paolo, Carolina
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Seiler, Thomas B.
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Hu, Meng
    Helmholtz-Zentrum für Umweltforschung (UFZ), Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Muz, Melis
    Helmholtz-Zentrum für Umweltforschung (UFZ), Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Brack, Werner
    Helmholtz-Zentrum für Umweltforschung (UFZ), Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany; College of Resources and Environmental Science, Chongqing University Beibei, Chongqing, China; College of Environmental Science and Engineering and State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai, China; State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing, China.
    The value of zebrafish as an integrative model in effect-directed analysis: a review2015In: Environmental Sciences Europe, ISSN 2190-4707, E-ISSN 2190-4715, Vol. 27, no 8, p. 1-11Article, review/survey (Refereed)
    Abstract [en]

    Bioassays play a central role in effect-directed analysis (EDA), and their selection and application have to consider rather specific aspects of this approach. Meanwhile, bioassays with zebrafish, an established model organism in different research areas, are increasingly being utilized in EDA. Aiming to contribute for the optimal application of zebrafish bioassays in EDA, this review provides a critical overview of previous EDA investigations that applied zebrafish bioassays, discusses the potential contribution of such methods for EDA and proposes strategies to improve future studies. Over the last 10 years, zebrafish bioassays have guided EDA of natural products and environmental samples. The great majority of studies performed bioassays with embryos and early larvae, which allowed small-scale and low-volume experimental setups, minimized sample use and reduced workload. Biotesting strategies applied zebrafish bioassays as either the only method guiding EDA or instead integrated into multiple bioassay approaches. Furthermore, tiered biotesting applied zebrafish methods in both screening phase as well as for further investigations. For dosing, most of the studies performed solvent exchange of extracts and fractions to dimethyl sulfoxide (DMSO) as carrier. However, high DMSO concentrations were required for the testing of complex matrix extracts, indicating that future studies might benefit from the evaluation of alternative carrier solvents or passive dosing. Surprisingly, only a few studies reported the evaluation of process blanks, indicating a need to improve and standardize methods for blank preparation and biotesting. Regarding evaluated endpoints, while acute toxicity brought limited information, the assessment of specific endpoints was of strong value for bioactivity identification. Therefore, the bioassay specificity and sensitivity to identify the investigated bioactivity are important criteria in EDA. Additionally, it might be necessary to characterize the most adequate exposure windows and assessment setups for bioactivity identification. Finally, a great advantage of zebrafish bioassays in EDA of environmental samples is the availability of mechanism- and endpoint-specific methods for the identification of important classes of contaminants. The evaluation of mechanism-specific endpoints in EDA is considered to be a promising strategy to facilitate the integration of EDA into weight-of-evidence approaches, ultimately contributing for the identification of environmental contaminants causing bioassay and ecological effects.

  • 12.
    Fassbender, Christopher
    et al.
    Aquatic Ecology and Toxicology Section, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Section, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research, (Biology V), RWTH Aachen University, Aachen, Germany.
    Gene-TEQ-a standardized comparative assessment of effects in the comet assay using genotoxicity equivalents2012In: Journal of Environmental Monitoring, ISSN 1464-0325, E-ISSN 1464-0333, Vol. 14, p. 1325-1334Article in journal (Refereed)
    Abstract [en]

    Existing methods for the comparison of genotoxic effects in the comet assay bear considerable disadvantages such as the problem to link information about concentration dependence and severity of effects. Moreover, given the lack of standardized protocols and the use of various standards, it may be extremely difficult to compare different studies. In order to provide a method for standardized comparative assessment of genotoxic effects, the concept of genotoxicity equivalents (Gene-TEQ) was developed. As potential reference compounds for genotoxic effects, three directly acting (N-methyl-N0-nitro-N-nitrosoguanidine (MNNG), methyl-methanesulfonate, and N-methyl-N-nitrosourea) and three indirectly acting (cyclophosphamide, dimethylnitrosamine, and 4-nitroquinoline-oxide) genotoxic substances were compared with respect to their cytotoxic (neutral red) and genotoxic (cometassay) concentration–response profiles in the permanent fish cell line RTL-W1. For further comparison, two sediment extracts from the upper Danube River were investigated as environmental samples. Based on the results of cytotoxicity and genotoxicity testing, MNNG was selected as the reference compound. At several exposure levels and durations, genotoxic effects of both the other pure substances and the environmental samples were calculated as percentages of the maximum MNNGeffect and related to the absolute MNNG effect (EC values). Thus, genotoxicity equivalent factors(Gene-TEQs) relative to MNNG could be calculated. Gene-TEQs can easily be applied to puresubstances, mixtures and field samples to provide information about their toxicity relative to the reference compound. Furthermore, the Gene-TEQ concept allows a direct comparison of environmental samples from different laboratories.

  • 13.
    Garcia-Käufer, Manuel
    et al.
    Hydrotox GmbH, Freiburg, Germany; Department of Ecosystem Analysis, Institute for Environmental Research, ABBT–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany; Center for Complementary Medicine, Department of Environmental Health Sciences, University Medical Centre Freiburg, Freiburg, Germany.
    Gartiser, Stefan
    Hydrotox GmbH, Freiburg, Germany.
    Hafner, Christoph
    Hydrotox GmbH, Freiburg, Germany.
    Schiwy, Sabrina
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBT–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBT–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Gruendemann, Christian
    Center for Complementary Medicine, Department of Environmental Health Sciences, University Medical Centre Freiburg, Freiburg, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBT–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany; School of Environment, Nanjing University, Nanjing, China; Key Laboratory of Yangtze River Environment of Education Ministry of China, College of Environmental Science and Engineering, Tongji University, Shanghai, China.
    Genotoxic and teratogenic effect of freshwater sediment samples from the Rhine and Elbe River (Germany) in zebrafish embryo using a multi-endpoint testing strategy2015In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 22, no 21, p. 16341-16357Article in journal (Refereed)
    Abstract [en]

    The embryotoxic potential of three model sediment samples with a distinct and well-characterized pollutant burden from the main German river basins Rhine and Elbe was investigated. The Fish Embryo Contact Test (FECT) in zebrafish (Danio rerio) was applied and submitted to further development to allow for a comprehensive risk assessment of such complex environmental samples. As particulate pollutants are constructive constituents of sediments, they underlay episodic source-sink dynamics, becoming available to benthic organisms. As bioavailability of xenobiotics is a crucial factor for ecotoxicological hazard, we focused on the direct particle-exposure pathway, evaluating through put-capable endpoints and considering toxicokinetics. Fish embryo and larvae were exposed toward reconstituted (freeze-dried) sediment samples on a microcosm-scale experimental approach. A range of different developmental embryonic stages were considered to gain knowledge of potential correlations with metabolic competence during the early embryogenesis. Morphological, physiological, and molecular endpoints were investigated to elucidate induced adverse effects, placing particular emphasis on genomic instability, assessed by the in vivo comet assay. Flow cytometry was used to investigate the extent of induced cell death, since cytotoxicity can lead to confounding effects. The implementation of relative toxicity indices further provides inter-comparability between samples and related studies. All of the investigated sediments represent a significant ecotoxicological hazard by disrupting embryogenesis in zebrafish. Beside the induction of acute toxicity, morphological and physiological embryotoxic effects could be identified in a concentration-response manner. Increased DNA strand break frequency was detected after sediment contact in characteristic non-monotonic dose–response behavior due to overlapping cytotoxic effects. The embryonic zebrafish toxicity model along with the in vivo comet assay and molecular biomarker analysis should prospectively be considered to assess the ecotoxicological potential of sediments allowing for a comprehensive hazard ranking. In order to elucidate mode of action, novel techniques such as flow cytometry have been adopted and proved to be valuable tools for advanced risk assessment and management.

  • 14.
    Grund, Stefanie
    et al.
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg Im Neuenheimer Feld, Heidelberg, Germany.
    Keiter, Steffen
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg Im Neuenheimer Feld, Heidelberg, Germany.
    Böttcher, Melanie
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg Im Neuenheimer Feld, Heidelberg, Germany.
    Seitz, Nadja
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg Im Neuenheimer Feld, Heidelberg, Germany.
    Wurm, Karl
    Gewässerökologisches Labor, Starzach, Germany.
    Manz, Werner
    Biochemistry/Ecotoxicology, German Federal Institute of Hydrology, Koblenz, Germany.
    Hollert, Henner
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg Im Neuenheimer Feld, Heidelberg, Germany; Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Section, Department of Zoology, University of Heidelberg Im Neuenheimer Feld, Heidelberg, Germany.
    Assessment of fish health status in the Upper Danube River by investigation of ultrastructural alterations in the liver of barbel Barbus barbus2010In: Diseases of Aquatic Organisms, ISSN 0177-5103, E-ISSN 1616-1580, Vol. 23, p. 235-248Article in journal (Refereed)
    Abstract [en]

    Despite intensive efforts and tightened guidelines for improvement of water quality over the last 2 decades, declines of fish populations have been reported for several rivers around the world. The present study forms part of a comprehensive weight-of-evidence approach, which aims to identify potential causes for the decline in fish catches observed in the Upper Danube River. The major focus of the present study is the investigation of the health status of wild barbel Barbus barbus L. collected from 3 locations along the Danube River, which experienced different levels of contamination. Whereas the comparison of the condition factor (CF) of field fish with that of control fish revealed no differences, ultrastructural investigations indicated severe disturbance of hepatic cell metabolism in field fish from the more contaminated sites Rottenacker and Ehingen, compared to both control fish and field fish from the less contaminated site Riedlingen. The ultrastructural analysis provided information about reactions of e.g. the rough endoplasmic reticulum, peroxisomes, andmitochondria, indicating an impaired health status of barbel at the sampling sites Rottenacker and Ehingen. Even though a straightforward cause-effect relationship between sediment contamination and ultrastructural alterations could not be established, based on a meta-analysis and toxicity assays it may be suggested that sediment-bound xenobiotics at least partly account for the hepatocellular changes. A relationship between impaired fish health status and the decline of fish catches along the Upper Danube River cannot be excluded.

  • 15.
    Gustavsson, Lillemor
    et al.
    Örebro University, School of Science and Technology. Karlskoga Environment and Energy Company, Karlskoga, Sweden.
    Heger, Sebastian
    Department of Ecosystem Analysis, Institute for Environmental, Research, RWTH Aachen University, Aachen, Germany.
    Ejlertsson, Jörgen
    Scandinavian Biogas Fuels AB, Stockholm, Sweden.
    Ribé, Veronica
    School of Sustainable Development of Society and Technology, Mälardalen University, Västerås, Sweden.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental, Research, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental, Research, RWTH Aachen University, Aachen, Germany.
    Industrial sludge containing pharmaceutical residues and explosives alters inherent toxic properties when co-digested with oat and post-treated in reed beds2014In: Environmental Sciences Europe, ISSN 2190-4707, E-ISSN 2190-4715, Vol. 26, no 8, p. 1-11Article in journal (Refereed)
    Abstract [en]

    Background: Methane production as biofuels is a fast and strong growing technique for renewable energy. Substrateslike waste (e.g. food, sludge from waste water treatment plants (WWTP), industrial wastes) can be used as a suitable resource for methane gas production, but in some cases, with elevated toxicity in the digestion residue. Former investigations have shown that co-digesting of contaminated waste such as sludge together with other substrates canproduce a less toxic residue. In addition, wetlands and reed beds demonstrated good results in dewatering and detoxifying of sludge. The aim of the present study was to investigate if the toxicity may alter in industrial sludgeco-digested with oat and post-treatment in reed beds. In this study, digestion of sludge from Bjorkborn industrial area in Karlskoga (reactor D6) and co-digestion of the same sludge mixed with oat (reactor D5) and post-treatment in reed beds were investigated in parallel. Methane production as well as changes in cytotoxicity (Microtox(R); ISO 11348–3), genotoxicity (Umu-C assay; ISO/13829) and AhR-mediated toxicity (7-ethoxyresorufin-O-deethylase (EROD) assay using RTW cells) were measured.

    Results: The result showed good methane production of industrial sludge (D6) although the digested residue was more toxic than the ingoing material measured using microtox30min and Umu-C. Co-digestion of toxic industrial sludge and oat(D5) showed higher methane production and significantly less toxic sludge residue than reactor D6. Furthermore, dewatering and treatment in reed beds showed low and non-detectable toxicity in reed bed material and outgoingwater as well as reduced nutrients.

    Conclusions: Co-digestion of sludge and oat followed by dewatering and treatment of sludge residue in reed beds canbe a sustainable waste management and energy production. We recommend that future studies should involve co-digestion of decontaminated waste mixed with different non-toxic material to find a substrate mixture that producethe highest biogas yield and lowest toxicity within the sludge residue.

  • 16.
    Hafner, Christoph
    et al.
    Hydrotox GmbH, Freiburg, Germany.
    Gartiser, Stefan
    Hydrotox GmbH, Freiburg, Germany.
    Garcia-Kaeufer, Manuel
    Hydrotox GmbH, Freiburg, Germany; Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Schiwy, Sabrina
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Hercher, Christoph
    Hydrotox GmbH, Freiburg, Germany.
    Meyer, Wiebke
    Institute of Geology and Palaeontology – Applied Geology, University of Münster, Münster, Germany.
    Achten, Christine
    Institute of Geology and Palaeontology – Applied Geology, University of Münster, Münster, Germany.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Investigations on sediment toxicity of German rivers applying a standardized bioassay battery2015In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 22, no 21, p. 16358-16370Article in journal (Refereed)
    Abstract [en]

    River sediments may contain a huge variety of environmental contaminants and play a key role in the ecological status of aquatic ecosystems. Contaminants adsorbed to sediments and suspended solids may contribute directly or after remobilization to an adverse ecological and chemical status of surface water. In this subproject of the joint research project DanTox, acetonic Soxhlet extracts from three German river sediments from the River Rhine (Altrip and Ehrenbreitstein with moderate contamination) and River Elbe (Veringkanal Hamburg heavily contaminated) were prepared and redissolved in dimethyl sulfoxide (DMSO). These extracts were analyzed with a standard bioassay battery with organisms from different trophic levels (bacteria, algae, Daphnia, fish) as well as in the Ames test and the umuC test for bacterial mutagenicity and genotoxicity according to the respective OECD and ISO guidelines. In total, 0.01 % (standard) up to 0.25 % (only fish embryo test) of the DMSO sediment extract was dosed to the test systems resulting in maximum sediment equivalent concentrations (SEQ) of 2 up to 50 g l(-1). The sediment of Veringkanal near Hamburg harbor was significantly more toxic in most tests compared to the sediment extracts from Altrip and Ehrenbreitstein from the River Rhine. The most toxic effect found for Veringkanal was in the algae test with an ErC50 (72 h) of 0.00226 g l(-1) SEQ. Ehrenbreitstein and Altrip samples were about factor 1,000 less toxic. In the Daphnia, Lemna, and acute fish toxicity tests, no toxicity at all was found at 2 g l(-1) SEQ. corresponding to 0.01 % DMSO. Only when increasing the DMSO concentration the fish embryo test showed a 22-fold higher toxicity for Veringkanal than for Ehrenbreitstein and Altrip samples, while the toxicity difference was less evident for the Daphnia test due to the overlaying solvent toxicity above 0.05 % dimethyl sulfoxide (DMSO). The higher toxicities observed with the Veringkanal sample are supported by the PAH and PCB concentrations analyzed in the sediments. The sediment extracts of Altrip andVeringkanal were mutagenic in the Ames tester strain TA98 with metabolic activation (S9mix). The findings allow a better ecotoxicological characterization of the sediments extensively analyzed in all subprojects of the DanTox project (e. g., Garcia-Kaeufer et al. Environ Sci Pollut Res. doi: 10.1007/s11356-014-3894-4, 2014; Schiwy et al. Environ Sci Pollut Res. doi: 10.1007/s11356-014-31850, 2014; Hollert and Keiter 2015). In the absence of agreed limit values for sediment extracts in standard tests, further data with unpolluted reference sediments are required for a quantitative risk assessment of the investigated polluted sediments.

  • 17.
    Hausen, Jonas
    et al.
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Otte, Jens C.
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany.
    Legradi, Jessica
    Environment and Health, Vrije Universiteit Amsterdam, Amsterdam, Netherlands.
    Yang, Lixin
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany .
    Strähle, Uwe
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany.
    Fenske, Martina
    Project Group for Translational Medicine and Pharmacology, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany .
    Hecker, Markus
    School of Environment and Sustainability, University of Saskatchewan, Saskatoon SK, Canada.
    Tang, Song
    School of Environment and Sustainability, University of Saskatchewan, Saskatoon SK, Canada.
    Hammers-Wirtz, Monika
    Research Institute for Ecosystem Analysis and Assessment [gaiac], Aachen, Germany.
    Hollert, Henner
    Institute for Environmental Research [RWTH], Aachen University, Aachen, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Institute for Environmental Research, RWTH Aachen University, Aachen, Germany; Man-Technology-Environment Research Centre, Örebro University, Örebro, Sweden.
    Ottermanns, Richard
    Institute for Environmental Research [RWTH], Aachen University, Aachen, Germany .
    Fishing for contaminants: identification of three mechanism specific transcriptome signatures using Danio rerio embryos2018In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 25, no 5, p. 4023-4036Article in journal (Refereed)
    Abstract [en]

    In ecotoxicology, transcriptomics is an effective way to detect gene expression changes in response to environmental pollutants. Such changes can be used to identify contaminants or contaminant classes and can be applied as early warning signals for pollution. To do so, it is important to distinguish contaminant-specific transcriptomic changes from genetic alterations due to general stress. Here we present a first step in the identification of contaminant class-specific transcriptome signatures. Embryos of zebrafish (Danio rerio) were exposed to three substances (methylmercury, chlorpyrifos and Aroclor 1254, each from 24 to 48 hpf exposed) representing sediment typical contaminant classes. We analyzed the altered transcriptome to detect discriminative genes significantly regulated in reaction to the three applied contaminants. By comparison of the results of the three contaminants, we identified transcriptome signatures and biologically important pathways (using Cytoscape/ClueGO software) that react significantly to the contaminant classes. This approach increases the chance of finding genes that play an important role in contaminant class-specific pathways rather than more general processes.

  • 18.
    Hausen, Jonas
    et al.
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Otte, Jens C.
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany.
    Straehle, Uwe
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany.
    Hammers-Wirtz, Monika
    Research Institute for Ecosystem Analysis and Assessment - gaiac, Aachen, Germany.
    Hollert, Henner
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen H.
    Örebro University, School of Science and Technology. Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Ottermanns, Richard
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Fold-change threshold screening: a robust algorithm to unmask hidden gene expression patterns in noisy aggregated transcriptome data2015In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 22, no 21, p. 16384-16392Article in journal (Refereed)
    Abstract [en]

    Transcriptomics is often used to investigate changes in an organism's genetic response to environmental contamination. Data noise can mask the effects of contaminants making it difficult to detect responding genes. Because the number of genes which are found differentially expressed in transcriptome data is often very large, algorithms are needed to reduce the number down to a few robust discriminative genes. We present an algorithm for aggregated analysis of transcriptome data which uses multiple fold-change thresholds (threshold screening) and p values from Bayesian generalized linear model in order to assess the robustness of a gene as a potential indicator for the treatments tested. The algorithm provides a robustness indicator (ROBI) as well as a significance profile, which can be used to assess the statistical significance of a given gene for different fold-change thresholds. Using ROBI, eight discriminative genes were identified from an exemplary dataset (Danio rerio FET treated with chlorpyrifos, methylmercury, and PCB) which could be potential indicators for a given substance. Significance profiles uncovered genetic effects and revealed appropriate fold-change thresholds for single genes or gene clusters. Fold-change threshold screening is a powerful tool for dimensionality reduction and feature selection in transcriptome data, as it effectively reduces the number of detected genes suitable for environmental monitoring. In addition, it is able to unmask patterns in altered genetic expression hidden by data noise and reduces the chance of type II errors, e.g., in environmental screening.

  • 19.
    Hollert, Henner
    et al.
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany; College of Environmental Science and Engineering and State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanhai, China; School of Environment, Nanjing University, Nanjing, China; College of Resources and Environmental Science, Chongqing University, Chongqing, China.
    Crawford, Sarah E.
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Brack, Werner
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany; Department Effect-Directed Analysis, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany.
    Brinkmann, Markus
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany; Toxicology Centre, University of Saskatchewan, Saskatoon, Canada.
    Fischer, Elske
    Laboratory for Archaeobotany, Landesamt für Denkmalpflege im Regierungspräsidium Stuttgart, Gaienhofen-Hemmenhofen, Germany.
    Hartmann, Kai
    Institute for Geographical Sciences, Freie Universität Berlin, Berlin, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Ottermanns, Richard
    Institute for Environmental Research (Biology V), Department of Environmental Biology and Chemodynamics, RWTH Aachen University, Aachen, Germany.
    Ouellet, Jacob D.
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Rinke, Karsten
    Department of Lake Research, Helmholtz Centre for Environmental Research - UFZ, Magdeburg, Germany.
    Rösch, Manfred
    Laboratory for Archaeobotany, Landesamt für Denkmalpflege im Regierungspräsidium Stuttgart, Gaienhofen-Hemmenhofen, Germany.
    Ross-Nickoll, Martina
    Institute for Environmental Research (Biology V), Department of Environmental Biology and Chemodynamics, RWTH Aachen University, Aachen, Germany.
    Schäffer, Andreas
    Institute for Environmental Research (Biology V), Department of Environmental Biology and Chemodynamics, RWTH Aachen University, Aachen, Germany.
    Schüth, Christoph
    Institute for Applied Geoscience, Technische Universität Darmstadt, Darmstadt, Germany; Institute of Geosystems and Bioindication, Technische Universität Braunschweig, Braunschweig, Germany.
    Schulze, Tobias
    Department Effect-Directed Analysis, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany.
    Schwarz, Anja
    Institute of Geosystems and Bioindication, Technische Universität Braunschweig, Braunschweig, Germany.
    Seiler, Thomas-Benjamin
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Wessels, Martin
    Institute for Lake Research, State Institute for Environment, Measurements and Nature Conservation Baden-Württemberg (LUBW), Langenargen, Germany.
    Hinderer, Matthias
    Institute for Applied Geoscience, Technische Universität Darmstadt, Darmstadt, Germany.
    Schwalb, Antje
    Institute of Geosystems and Bioindication, Technische Universität Braunschweig, Braunschweig, Germany.
    Looking back - Looking forward: A novel multi-time slice weight-of-evidence approach for defining reference conditions to assess the impact of human activities on lake systems2018In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 626, p. 1036-1046Article in journal (Refereed)
    Abstract [en]

    Lake ecosystems are sensitive recorders of environmental changes that provide continuous archives at annual to decadal resolution over thousands of years. The systematic investigation of land use changes and emission of pollutants archived in Holocene lake sediments as well as the reconstruction of contamination, background conditions, and sensitivity of lake systems offer an ideal opportunity to study environmental dynamics and consequences of anthropogenic impact that increasingly pose risks to human well-being. This paper discusses the use of sediment and other lines of evidence in providing a record of historical and current contamination in lake ecosystems. We present a novel approach to investigate impacts from human activities using chemical-analytical, bioanalytical, ecological, paleolimnological, paleoecotoxicological, archeological as well as modeling techniques. This multi-time slice weight-of-evidence (WOE) approach will generate knowledge on conditions prior to anthropogenic influence and provide knowledge to (i) create a better understanding of the effects of anthropogenic disturbances on biodiversity, (ii) assess water quality by using quantitative data on historical pollution and persistence of pollutants archived over thousands of years in sediments, and (iii) define environmental threshold values using modeling methods. This technique may be applied in order to gain insights into reference conditions of surface and ground waters in catchments with a long history of land use and human impact, which is still a major need that is currently not yet addressed within the context of the European Water Framework Directive.

  • 20.
    Hollert, Henner
    et al.
    Universität Heidelberg, Institut für Zooiogie, Heidelberg, Germany .
    Heise, Susanne
    BIS Beratungszentrum für Integriertes Sedimentmanagement an der TU Hamburg-Harburg, Hamburg-Harburg, Germany.
    Keiter, Steffen
    Universität Heidelberg, Institut für Zooiogie, Heidelberg, Germany .
    Heininger, Peter
    Bundesanstalt für Gewässerkunde, Koblenz, Germany.
    Förstner, Ulrich
    Technische Universität Hamburg-Harburg, Institut für Umwelttechnik und Energiewirtschaft, Hamburg-Harburg, Germany.
    Wasserrahmenrichtlinie: Fortschritte und Defizite2007In: Umweltwissenschaften und Schadstoff-Forschung, ISSN 0934-3504, E-ISSN 1865-5084, Vol. 19, no 1 Suppl., p. 58-70Article in journal (Refereed)
    Abstract [en]

    Background: The water protection policy of the European Union sits on new footings since the end of 2000: The Water Framework

    Directive (WFD). By replacing, merging and renewing all parts of the European water protection policy from the 1970s, the WFD provides a consistent, transparent and comprehensive concept of what water management should be in the Europe of the coming decades. The new directive is aimed at a holistic approach towards integrated water protection. It sets ambitious high-quality goals to achieve a good status for European lakes and rivers primarily in ecological terms, gives details about the essential processes as well as instruments, and includes everything into a strict time schedule.

    Aim: This article adresses progress and shortcomings at the implementation of the WFD in general and with reference to two selected case studies (Rivers Elbe and Upper Danube).

    Results and Discussion: After introducing the WFD, its aims and exceptions, a policy summary and background document ‘Environmental objectives und the Water Framework Directive’ and the use of Environmental Quality Standards (EQS) for single ‘priority substances’ as well as ‘hazardous priority components’ is discussed. The initial characterization undertaken by the German states revealed that only about 14% of all surface waters are considered to meet the WFD objectives by the year of 2015. Approximately 60% of the water bodies assessed are at risk of failing the WFD objectives, if not systematic efforts are made to improve the quality. Screenings of sources and paths of exposure for ‘priority substances’ and ‘priority hazardous substances’ according WFD identified one distinct pollution source for surface waters: ‘Historical pollution from sediments’. Because of industrial emissions in the past several river catchment areas are expected to fail the standards demanded by the WFD, due to a risk of remobilization of contaminants from sediments. This holds true for the Rhine river with high loads of hexachlorobenzene (HCB) as well as for Elbe river, where contaminated sediments can be a severe problem. Therefore, integration of sediments into the holistic river basin management approach and their consideration within the ‘programmes of measures’ scheduled for 2009 is highly recommended. At present, a comprehensive weight-of-evidence study verifies whether the observed fish decline at the Upper Danube. River is caused by ecotoxicological hazard potentials of contaminated sediments.

    Outlook: Combined investigations of sediment contamination and mobility as well as acute and mechanism specific biotests in effect directed analyses/weight-of-evidence studies show grent potential for the assessment of chemically polluted rivers and should be included into the ‘programmes of measures’ within future management concepts.

  • 21. Hollert, Henner
    et al.
    Keiter, Steffen
    Böttcher, Melanie
    Grund, Steffi
    Seitz, Nadja
    Otte, Jens
    Bluhm, Kerstin
    Wurm, Karl
    Hecker, Markus
    Higley, Eric
    Giesy, John
    Takner [Olsman], Helena
    van Bavel, Bert
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Reifferscheid, Georg
    Manz, Werner
    Erdinger, Lother
    Schulze, Tobias
    Luebcke-van Varel, Urte
    Kammann, Ulrike
    Schöneberger, René
    Suter, Marc
    Brack, Werner
    Strähle, Uwe
    Braunbeck, Thomas
    Eine Weight-of-Evidence-Studie zur Bewertung der Sedimentbelastung und des Fischrückgangs in der Oberen Donau [Assessing sediments and fish health using a weight-of-evidence approach : in search for the causes of fish decline in the Danube river]2009In: Umweltwissenschaften und Schadstoff-Forschung, ISSN 0934-3504, E-ISSN 1865-5084, Vol. 21, no 3, p. 260-263Article in journal (Refereed)
    Abstract [en]

    Background and aim Despite intensive and continuous stocking and improvement of water quality since the 1970s, fish populations, especially those of the grayling (Thymallus thymallus), have declined over the last two decades in the upper Danube River (Germany). In order to assess 1) possible links between molecular/biochemical responses and ecologically relevant effects, and 2) if ecotoxicological effects might be related to the decline in fish catches in the upper Danube river, sediment samples and fish were collected at different locations and analyzed using a weight-of-evidence (WOE) approach with several lines of evidence. The objective of the presentation is to introduce the conceptual framework and to review results of the ongoing study. As previously addressed by Chapman and Hollert (2006) a variety of lines of evidence can be used in WOE studies. Briefly, 1) a comprehensive battery of acute and mechanism-specific bioassays was used to characterize the ecotoxicological hazard potential. 2) Histopathological investigations and the micronucleus assay with erythrocytes were applied, analyzing in situ parameters. 3) Diversity and abundance of benthic macroinvertebrates and fish as well as 4) persistent organic pollutants, endocrine disrupting substances, limnochemical parameters and the concentration of heavy metals were recorded. To identify organic contaminants a spotential causes of sediment toxicity assays, 5) effect directed analysis was applied. © 2009 Springer-Verlag.

  • 22.
    Hollert, Henner
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Achen, Germany.
    Keiter, Steffen H.
    Örebro University, School of Science and Technology. Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Achen, Germany.
    Danio rerio as a model in aquatic toxicology and sediment research2015In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 22, p. 16243-16246Article in journal (Refereed)
  • 23.
    Hollert, Henner
    et al.
    University of Heidelberg.
    Keiter, Steffen
    University of Heidelberg.
    König, Natalie
    University of Heidelberg.
    Rudolf, Marc
    University of Heidelberg.
    Ulrich, Markus
    University of Heidelberg.
    Braunbeck, Thomas
    University of Heidelberg.
    A new sediment contact assay to assess particle-bound pollutants using zebrafish (Danio rerio) embryos2003In: Journal of Soils and Sediments, ISSN 1439-0108, E-ISSN 1614-7480, Vol. 3, no 3, p. 197-207Article in journal (Refereed)
    Abstract [en]

    Goal, Scope and Background. Based on a bioassay battery coveringonly primary producers and consumers as well as degraders, the potential ecological hazard of sediments to vertebrates cannot be estimated comprehensively. Therefore, there is an urgent need to develop and standardize integrated vertebrate-based test systems for sediment investigation strategies. Whereas vertebrate-based in vitro systems have frequently been used for the investigation of aqueous samples, there is a significant lack of whole sediment assays. Thus, the purpose of the present study was: (1)to develop a rapid and reliable, but comprehensive method to investigate native sediments and particulate matters without preceding extraction procedures; (2) to compare the hazard potential of solid phase sediments to the effects of corresponding pore waters and organic extracts in order to characterize the bioavailability of the particle-bound pollutants; and (3) to relatively evaluate the embryotoxic effects of sediments from the catchment areas of the rivers Rhine, Neckar and Danube.

    Methods (or Main Features). To investigate the toxicity of sediment samples on vertebrates, the standard embryo toxicity test with the zebrafish (Danio rerio; Hamilton-Buchanan 1922) according to DIN 38415-6 was modified with respect to exposure scheme and toxicological endpoints. Sediments from the catchment area of the Neckar River were assessed using pore waters, acetonic extracts and native sediments in order to get inside into the potential bioavailability of particle-bound pollutants. A comprehensive test protocol for the investigation of native sediments in the embryo toxicity test with the zebrafish is presented.

    Results and Discussion. The fish embryo assay with Danio rerio can be carried out with both aqueous and organic sediment extracts as well as native (whole, solid phase) sediment samples. Elongation of exposure time from 48 to up to 196 h significantly increased the mortality. Using the fish egg assay with native sediments, a broad range of embryotoxic effects could be elucidated, including clear-cut dose-response curves for the embryotoxic effects of contaminated sediments; in contrast, absence ofembryotoxic effects could be demonstrated even for the highest test concentrations of unpolluted sediments. With native sediments, embryotoxicity was clearly higher than with corresponding pore waters, thus corroborating the view that – at least for fish eggs – the bioavailability of particle-bound lipophilic substances in native sediments is higher than generally assumed. The relative ranking of sediment toxicity was identical using both native sediments and sediment extracts, EC20 values of the latter, however, being eight time lower higher than with the native sediments. A comparison of the embryo toxic effects of samples from the Neckar area with locations along the Rhine and Danube rivers elucidated a broad range of results, thus indicating different levels of contamination.

    Conclusions. A modified protocol of the zebrafish embryo test allows the assessment of sediment toxicity in both aqueous extracts and native sediments. The isolated investigation of pore waters may result in a clear-cut underestimation of the bioavailability of lipophilic particle-bound substances (as determined by native sediments).

    Recommendations and Perspectives. The zebrafish embryo test with native (whole, solid phase) sediments appears very promising for the evaluation of the bioavailable fraction of lipophilicparticle-bound substances and can therefore be recommended for the evaluation of vertebrate toxicity in tiered sediment test strategies and dredging directives such as the HABAB-WSV. Whereas acetone extracts may be tested as a rough estimation of embryotoxicity, native sediment samples will provide a more comprehensive and realistic insight into the bioavailable hazard potential

  • 24.
    Jernbro, Susanne
    et al.
    Örebro University, Department of Natural Sciences. University of Heidelberg, Heidelberg, Germany.
    Suares Rocha, Paula
    University of Heidelberg, Heidelberg, Germany.
    Keiter, Steffen
    University of Heidelberg, Heidelberg, Germany.
    Skutlarek, Dirk
    University of Bonn, Germany.
    Färber, Harald
    University of Bonn, Germany.
    Jones, Paul D.
    Michigan State University, Michigan, USA.
    Giesy, John P.
    Michigan State University, Michigan, USA.
    Hollert, Henner
    University of Heidelberg, Heidelberg, Germany.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Perfluorooctane Sulfonate Increases the Genotoxicity of Cyclophosphamide in the Micronucleus Assay with V79 Cells: Further Proof of Alterations in Cell Membrane Properties Caused by PFOS2007In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 14, no 2, p. 85-87Article in journal (Refereed)
    Abstract [en]

    Perfluorooctane sulfonate (PFOS; C8F17SO3–) is a fully fluorinated organic compound which has been manufactured for decades and was used widely in industrial and commercial products. The recent toxicological knowledge of PFOS mainly concerns mono-substance exposures of PFOS to biological systems, leaving the potential interactive effects of PFOS with other compounds as an area where understanding is significantly lacking. However, a recent study, reported the potential of PFOS to enhance the toxicity of two compounds by increasing cell membrane permeability. This is of particular concern since PFOS has been reported to be widely distributed in the environment where contaminants are known to occur in complex mixtures. In this study, PFOS was evaluated alone and in combination with cyclophosphamide (CPP) to investigate whether a presence of PFOS leads to an increased genotoxic potential of CPP towards hamster lung V79 cells. Genotoxicity was investigated using the micronucleus(MN) assay according to the recent draft ISO/DIS 21427-2 method. PFOS alone demonstrated no genotoxicity up to a concentration of 12.5 μg/ml. However, PFOS combined with two different concentrations of CPP, with metabolic activation, caused a significant increase in the number of micronucleated cells compared to treatments with CPP alone. These results provide a first indication that PFOS has the potential to enhance the genotoxic action of CPP towards V79 cells, suggesting, together with the alterations in cell membrane properties shown previously, that genotoxicity of complex mixtures may be increased significantly by changes in chemical uptake. Together with an earlier study performed by the own working group, it can be concluded that PFOS alone is not genotoxic in this bioassay using V79 cells up to 12.5 μg/ml, but that further investigations are needed to assess the potential interaction between PFOS and other substances, in particular regarding the impact of membrane alterations on the uptake of toxic substances.

  • 25.
    Kais, Britta
    et al.
    Aquatic Ecology and Toxicology Group, Center for Organismal Studies (COS), University of Heidelberg, Heidelberg, Germany.
    Schiwy, Sabrina
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology. Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Group, Center for Organismal Studies (COS), University of Heidelberg, Heidelberg, Germany.
    In vivo EROD assays with the zebrafish (Danio rerio) as rapid screening tools for the detection of dioxin-like activity2017In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 590-591, p. 269-280Article in journal (Refereed)
    Abstract [en]

    The present study compares two alternative in vivo approaches for the measurement of ethoxyresorufin-Odeethylase (EROD) activity in zebrafish (Danio rerio) following exposure to acetonic model sediment extracts: (1) the live-imaging EROD assay for the direct detection of EROD induction in individual livers via epifluorescence, and (2) the fish embryo EROD assay in subcellular fractions derived from entire zebrafish embryos after in vivo exposure.

    For toxicity assessment, each sediment extract was tested with the standard fish embryo test (FET). Upon completion of a functioning liver after 72 h, the embryos gave a distinct fluorescent signal in the liver, and a corresponding EROD activity could be detected in the fish embryo EROD assay. The exposure time in the live-imaging EROD assay was reduced to 3 h, which resulted in a stronger, less variable and more sensitive EROD response. Overall, the live-imaging and the fish embryo EROD assays showed the same tendencies and gave comparable results, e.g. a concentration-dependent increase in EROD activity at concentrations one order of magnitude below concentrations producing macroscopically visible abnormalities. At higher concentrations, however, a decrease of EROD activity was observed in either test. Both tests ranked the three model sediment extracts in the same order. Results indicate that both test systems complement each other and together provide a rapid and reliable in vivo tool to investigate the presence of dioxin-like substances in environmental samples.

  • 26.
    Karami, Ali
    et al.
    Department of Aquaculture, Faculty of Agriculture, University Putra Malaysia, Selangor, Malaysia.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Aachen, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Aachen, Germany.
    Courtenay, Simon
    Fisheries and Oceans Canada at the Canadian Rivers Institute, Department of Biology, University of New Brunswick, Fredericton NB, Canada.
    Fuzzy logic and adaptive neuro-fuzzy inference system for characterization of contaminant exposure through selected biomarkers in African catfish2013In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 20, no 3, p. 1586-1595Article in journal (Refereed)
    Abstract [en]

    This study represents a first attempt at applying a fuzzy inference system (FIS) and an adaptive neuro-fuzzy inference system (ANFIS) to the field of aquatic biomonitoring for classification of the dosage and time of benzo[a]pyrene (BaP) injection through selected biomarkers in African catfish (Clarias gariepinus). Fish were injected either intramuscularly (i.m.) or intraperitoneally (i.p.) with BaP. Hepatic glutathione S-transferase (GST) activities, relative visceral fat weights (LSI), and four biliary fluorescent aromatic compounds (FACs) concentrations were used as the inputs in the modeling study. Contradictory rules in FIS and ANFIS models appeared after conversion of bioassay results into human language (rule-based system). A “data trimming”approach was proposed to eliminate the conflicts prior to fuzzification. However, the model produced was relevant only to relatively low exposures to BaP, especially through the i.m. route of exposure. Furthermore, sensitivity analysis was unable to raise the classification rate to an acceptable level. In conclusion, FIS and ANFIS models have limited applications in the field of fish biomarker studies.

  • 27.
    Keiter, Steffen
    et al.
    Department of Zoology, Aquatic Ecology and Toxicology Section, University of Heidelberg, Heidelberg, Germany.
    Braunbeck, Thomas
    Department of Zoology, Aquatic Ecology and Toxicology Section, University of Heidelberg, Heidelberg, Germany.
    Heise, Susanne
    Institut für Biogefahrenstoffe und Umwelttoxikologie, Hamburg University of Applied Sciences, Hamburg, Germany.
    Pudenz, Stefan
    Westlakes Scientific Consulting Ltd, Department of Environmental Science, Cumbria, UK.
    Manz, Werner
    German Federal Institute of Hydrology, Koblenz, Germany.
    Hollert, Henner
    Department of Zoology, Aquatic Ecology and Toxicology Section, University of Heidelberg, Heidelberg, Germany; Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    A fuzzy logic-classification of sediments based on data from in vitro biotests2009In: Journal of Soils and Sediments, ISSN 1439-0108, E-ISSN 1614-7480, Vol. 9, no 3, p. 168-179Article in journal (Refereed)
    Abstract [en]

    Background, aim, and scope: Ecotoxicological risk assessment of sediments is usually based on a multitude of data obtained from tests with different endpoints. In the present study, a fuzzy logic-based model was developed in order to reduce the complexity of these data sets and to classify sediments on the basis of results from a battery of in vitro biotests.

    Materials and methods: The membership functions were adapted to fit the specific sensitivity and variability of each biotest. For this end, data sets were categorized into three toxicity levels using the box plot and empirical methods. The variability of each biotest was determined to calculatethe range of the gradual membership. In addition, the biotests selected were ranked according to the biological organisation level in order to consider the ecological relevance of the endpoints measured by selected over- or underestimation of the toxicity levels. In the next step of the fuzzy logic model, a rule-base was implemented using if...and...then decisions to arrive at a system of five quality classes.

    Results: The results of the classification of sediments fromthe Rhine and Danube Rivers showed the highest correlation between the biotest results and the fuzzy logical ternative based on the empirical method (i.e. the classification of the data sets into toxicity levels).

    Discussion: Many different classification systems based on biological test systems are depending on respective datasets; therefore, they are difficult to compare with other locations. Furthermore, they don‘t consider the inherent variability of biotests and the ecological relevance of these test systems as well. In order to create a comprehensive risk assessment for sediments, mathematical models should be used which take uncertainties of biotest systems into account, since they are of particular importance for areliable assessment. In the present investigation, the variability and ecological relevance of biotests were incorporated into a classification system based on fuzzy logic. Furthermore, since data from different sites and investigations were used to create membership functions ofthe fuzzy logic, this classification system has the potential to be independent of locations.

    Conclusions: In conclusion, the present fuzzy logic classification model provides an opportunity to integrate expert knowledge as well as acute and mechanism-specific effects for the classification of sediments for an ecotoxicological risk assessment.

    Recommendations and perspectives: In order to achieve amore comprehensive classification, further investigation is needed to incorporate results of chemical analyses and in situ parameters. Furthermore, more discussions are necessary with respect to the relative weight attributed to different ecological and chemical parameters in order too btain a more precise assessment of sediments.

  • 28. Keiter, Steffen
    et al.
    Grund, Stefanie
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Hagberg, Jessika
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Kammann, Ulrike
    Klempt, Martin
    Manz, Werner
    Olsman Takner, Helena
    Örebro University, Department of Natural Sciences.
    Braunbeck, Thomas
    Hollert, Henner
    Activities and identification of aryl hydrocarbon receptor agonists in sediments from the Danube river2008In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 390, no 8, p. 2009-2019Article in journal (Refereed)
    Abstract [en]

    This study is a consequence of a distinct fish decline in the Danube river since the beginning of the 1990s. In contrast to the decline of fish population, former studies have repeatedly documented that the water quality along the Danube river is improving. However, the conclusion of a pilot study in 2002 was that a high hazard potential is associated with local sediments. The present study documents that sediment samples from the Danube river showed comparatively high aryl hydrocarbon receptor mediated activity in biotests, using the cell lines GPC.2D.Luc, H4IIE (DR-CALUX®) and RTL-W1. The combination of chemical analysis, fractionation techniques and different in vitro tests revealed that priority pollutants could not explain the main induction, even though the concentrations of priority polycyclic aromatic hydrocarbons (PAHs) were very high (maximum in the tributary Schwarzach, sum of 16 EPA PAHs 26 μg/g). In conclusion, this investigation shows that nonpriority pollutants mainly mediate the high induction rates. Nevertheless, owing to the effects of PAHs towards fish and the connection between dioxin-like activity and carcinogenicity, the link between contamination and the fish population decline cannot be ruled out.

  • 29.
    Keiter, Steffen
    et al.
    University of Heidelberg, Heidelberg, Germany.
    Grund, Stefanie
    University of Heidelberg, Heidelberg, Germany.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Hagberg, Jessika
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Ulrike, Kammann
    Federal Research Centre for Fisheries, Hamburg, Germany.
    Klempt, Martin
    Federal Research Centre for Nutrition and Food, Hamburg, Germany.
    Manz, Werner
    Federal Institute of Hydrology, Koblenz, Germany.
    Olsman, Helena
    Örebro University, Department of Natural Sciences.
    Braunbeck, Thomas
    University of Heidelberg, Heidelberg, Germany.
    Hollert, Henner
    RWTH Aachen University, Aachen, Germany .
    Activities and identification of aryl hydrocarbon receptor agonists in sediments from the Danube river2008In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 390, no 8, p. 2009-2019Article in journal (Refereed)
    Abstract [en]

    This study is a consequence of a distinct fishdecline in the Danube river since the beginning of the 1990s. In contrast to the decline of fish population, former studies have repeatedly documented that the water quality along the Danube river is improving. However, the conclusion of a pilot study in 2002 was that a high hazard potentialis associated with local sediments. The present study documents that sediment samples from the Danube river showed comparatively high aryl hydrocarbon receptor mediated activity in biotests, using the cell lines GPC.2D.Luc, H4IIE(DR-CALUX®) and RTL-W1. The combination of chemical analysis, fractionation techniques and different in vitro tests revealed that priority pollutants could not explain the main induction, even though the concentrations of prioritypolycyclic aromatic hydrocarbons (PAHs) were very high (maximum in the tributary Schwarzach, sum of 16 EPAPAHs 26 μg/g). In conclusion, this investigation shows that nonpriority pollutants mainly mediate the high inductionrates. Nevertheless, owing to the effects of PAHs towards fish and the connection between dioxin-like activity andcarcinogenicity, the link between contamination and the fish population decline cannot be ruled out.

  • 30.
    Keiter, Steffen
    et al.
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Peddinghaus, Sabrina
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    Feiler, Ute
    German Federal Institute for Hydrology (BfG), Koblenz, Germany.
    von der Goltz, Britta
    Aquatic Ecology and Toxicology Group, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Hafner, Christoph
    Hydrotox GmbH, Freiburg, Germany.
    Ho, Nga Yu
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany.
    Rastegar, Sepand
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany.
    Otte, Jens
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany.
    Ottermanns, Richard
    Research Institute GAIAC, Aachen, Germany.
    Reifferscheid, Georg
    German Federal Institute for Hydrology (BfG), Koblenz, Germany.
    Strähle, Uwe
    Karlsruhe Institute of Technology (KIT), Institute of Toxicology and Genetics (ITG), Eggenstein-Leopoldshafen, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Group, Department of Zoology, University of Heidelberg, Heidelberg, Germany.
    Hammers-Wirtz, Monika
    Research Institute GAIAC, Aachen, Germany.
    Hollert, Henner
    Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Aachen, Germany.
    DanTox-a novel joint research project using zebrafish (Danio rerio) to identify specific toxicity and molecular modes of action of sediment-bound pollutants2010In: Journal of Soils and Sediments, ISSN 1439-0108, E-ISSN 1614-7480, Vol. 10, no 4, p. 714-717Article in journal (Refereed)
    Abstract [en]

    Introduction: The European Water Framework Directive aimsto achieve a good ecological and chemical status in surface water of European rivers by the year 2015. Since sediments and particulate matter act as secondary sources for pollutants, applied sediment toxicology is perceived to play a major rolefor obtaining new knowledge that can contribute to successful attainment of the goal. However, the existing bioassays for sediment toxicity analyses do not provide sufficient data concerning bioavailability of environmental pollutants. In this regard, there is an urgent need to combine sediment contact assays with gene expression analysis to investigate mechanism-specific sediment toxicity.

    Purpose: The aim of the novel joint research project is todevelop a eukaryotic test system, which can be used to investigate the ecotoxicological effects of contaminated sediments on gene expression level (DNA-array and RT-PCR).Current ecotoxicological research customarily involves a battery of bioassays to cover different toxicological endpoints (e.g., teratogenicity, genotoxicity, mutagenicity, Ah-receptor mediated toxicity, neurotoxicity). In contrast, methods that detect alterations in gene expression offer deeper insight by elucidating how chemical exposure and/or environmental challenge affect multiple metabolic pathways leading to these particular kinds of toxic response. Gene expression profiles reflect the way cells and organisms adapt or respond to a changing environment.

    Conclusion: The present project aspires to increase the fundamental molecular and physiological knowledge concerning the mode of action of environmental toxicants in zebrafish (Danio rerio). By working with partners from the academic and research institutions as well as from industry and waterway regulations, the success of this basic research-driven joint project in terms of development and implementation of novel sediment toxicity methods will be realized.

  • 31.
    Keiter, Steffen
    et al.
    Örebro University, School of Science and Technology.
    Rastall, Andrew
    University of Heidelberg, Heidelberg, Germany.
    Kosmehl, Thomas
    University of Heidelberg, Heidelberg, Germany.
    Wurm, Karl
    Gewässerökologisches Labor Starzach, The Eberhard Karls Universität Tübingen, Tübingen, Germany.
    Erdinger, Lothar
    University of Heidelberg, Heidelberg, Germany.
    Braunbeck, Thomas
    University of Heidelberg, Heidelberg, Germany.
    Hollert, Henner
    University of Heidelberg, Heidelberg, Germany.
    Ecotoxicological assessment of sediment, suspended matter andwater samples in the upper danube river: a pilot study in search for the causes for the decline of fish catches2006In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 13, no 5, p. 308-319Article in journal (Refereed)
    Abstract [en]

    Goals, Scope and Background. Fish populations, especially thoseof the grayling (Thymallus thymallus), have declined over thelast two decades in the upper Danube River between Sigmaringenand Ulm, despite intensive and continuous stocking and improvementof water quality since the 1970s. Similar problems havebeen reported for other rivers, e.g. in Switzerland, Great Britain,the United States and Canada. In order to assess if ecotoxicologicaleffects might be related to the decline in fish catchat the upper Danube River, sediment, suspended matter andwaste water samples from sewage treatment plants were collectedat selected locations and analyzed in a bioanalytical approachusing a battery of bioassays. The results of this pilotstudy will be used to decide if a comprehensive weight-of-evidencestudy is needed.

    Methods. Freeze-dried sediments and suspended particulate matterswere extracted with acetone in a Soxhlet apparatus. Organicpollutants from sewage water were concentrated usingXAD-resins. In order to investigate the ecotoxicological burden,the following bioassays were used: (1) neutral red assaywith RTL-W1 cells (cytotoxicity), (2) comet assay with RTLW1cells (genotoxicity), (3) Arthrobacter globiformis dehydrogenaseassay (toxicity to bacteria), (4) yeast estrogen screen assay(endocrine disruption), (5) fish egg assay with the zebrafish(Danio rerio; embryo toxicity) and (6) Ames test with TA98(mutagenicity).

    Results and Discussion. The results of the in vitro tests elucidateda considerable genotoxic, cytotoxic, mutagenic, bacteriotoxic,embryotoxic and estrogenic burden in the upper DanubeRiver, although with a very inhomogeneous distribution of effects.The samples taken from Riedlingen, for example, inducedlow embryo toxicity, but the second highest 17β-estradiol equivalentconcentration (1.8 ng/L). Using the fish egg assay with nativesediments, a broad range of embryotoxic effects could beelucidated, with clear-cut dose-response relationships for theembryotoxic effects of contaminated sediments. With nativesediments, embryotoxicity was clearly higher than with correspondingpore waters, thus corroborating the view that – atleast for fish eggs – the bioavailability of particle-bound lipophilicsubstances in native sediments is higher than generally assumed. The effect observed most frequently in the fish egg assay was a developmental delay. A comparison of our own results with locationsalong the rivers Rhine and Neckar demonstrated similaror even higher ranges of ecotoxicological burdens in theDanube River.

    Conclusions. The complex pattern of ecotoxicological effectscaused by environmental samples from the Danube River, whenassessed in an in vitro biotest battery using both acute and morespecific endpoints, showed that integration of different endpointsis essential for appropriate hazard assessment. Overall, theecotoxicological hazard potential shown has indeed to be consideredas one potential reason for the decline in fish catches atthe upper Danube River. However, based on the results of thispilot study, it is not possible to elucidate that chemically inducedalterations are responsible for the fish decline.

    Recommendations and Perspectives. In order to confirm the ecologicalrelevance of the in vitro results for the situation in thefield and especially for the decline of the grayling and otherfishes, further integrated investigations are required. For linkingthe weight of evidence obtained by in vitro assays and fishpopulation investigations, the application of additional, morespecific biomarkers (e.g. vitellogenin induction, EROD and micronucleusassay) has been initiated in fish taken from the fieldas well as in situ investigations.

  • 32.
    Keiter, Susanne
    et al.
    Aquatic Ecology and Toxicology Group, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Burkhard-Medicke, Kathleen
    Department Bioanalytical Ecotoxicology, UFZ-Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Wellner, Peggy
    Department Bioanalytical Ecotoxicology, UFZ-Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Kais, Britta
    Aquatic Ecology and Toxicology Group, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Färber, Harald
    Institute for Hygiene and Public Health, University of Bonn, Bonn, Germany.
    Skutlarek, Dirk
    Institute for Hygiene and Public Health, University of Bonn, Bonn, Germany.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Group, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Luckenback, Till
    Department Bioanalytical Ecotoxicology, UFZ-Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Does perfluorooctane sulfonate (PFOS) act as chemosensitizer in zebrafish embryos?2016In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 548-549, p. 317-324Article in journal (Refereed)
    Abstract [en]

    Earlier studies have shown that perfluorooctane sulfonate (PFOS) increases the toxicity of other chemicals by enhancing their uptake by cells and tissues. The present study aimed at testing whether the underlying mechanism of enhanced uptake of chemicals by zebrafish (Danio rerio) embryos in the presence of PFOS is by interference of this compound with the cellular efflux transporter Abcb4. Modifications of uptake/clearance and toxicity of two Abcb4 substrates, the fluorescent dye rhodamine B (RhB) and vinblastine, by PFOS were evaluated using 24 and 48. h post-fertilization (hpf) embryos. Upon 90. min exposure of 24. hpf embryos to 1. μM RhB and different PFOS concentrations (3-300. μM) accumulation of RhB in zebrafish was increased by up to 11.9-fold compared to controls, whereas RhB increases in verapamil treatments were 1.7-fold. Co-administration of PFOS and vinblastine in exposures from 0 to 48. hpf resulted in higher vinblastine-caused mortalities in zebrafish embryos indicating increased uptake of this compound. Interference of PFOS with zebrafish Abcb4 activity was further studied using recombinant protein obtained with the baculovirus expression system. PFOS lead to a concentration-dependent decrease of the verapamil-stimulated Abcb4 ATPase activity; at higher PFOS concentrations (250, 500. μM), also the basal ATPase activity was lowered indicating PFOS to be an Abcb4 inhibitor. In exposures of 48. hpf embryos to a very high RhB concentration (200. μM), accumulation of RhB in embryo tissue and adsorption to the chorion were increased in the presence of 50 or 100. μM PFOS. In conclusion, the results indicate that PFOS acts as inhibitor of zebrafish Abcb4; however, the exceptionally large PFOS-caused effect amplitude of RhB accumulation in the 1. μM RhB experiments and the clear PFOS effects in the experiments with 200. μM RhB suggest that an additional mechanism appears to be responsible for the potential of PFOS to enhance uptake of Abcb4 substrates.

  • 33.
    Kosmehl, Thomas
    et al.
    Aquatic Ecology and Toxicology Group, COS–Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Otte, Jens
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
    Yang, Lixin
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
    Legradi, Jessica
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
    Bluhm, Kerstin
    Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Aachen, Germany.
    Zinsmeister, Christian
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Aachen, Germany.
    Reifferscheid, Georg
    German Federal Institute for Hydrology (BfG), Koblenz, Germany.
    Manz, Werner
    Institute of Integrated Natural Sciences, University of Koblenz-Landau, Koblenz, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology Group, COS–Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Strähle, Uwe
    Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research (Biology V), RWTH Aachen University, Aachen, Germany.
    A combined DNA-microarray and mechanism-specific toxicity approach with zebrafish embryos to investigate the pollution of river sediments2012In: Reproductive Toxicology, ISSN 0890-6238, E-ISSN 1873-1708, Vol. 33, no 2, p. 245-253Article in journal (Refereed)
    Abstract [en]

    The zebrafish embryo has repeatedly proved to be a useful model for the analysis of effects by environmental toxicants. This proof-of-concept study was performed to investigate if an approach combining mechanism-specific bioassays with microarray techniques can obtain more in-depth insights into theecotoxicity of complex pollutant mixtures as present, e.g., in sediment extracts. For this end, altered gene expression was compared to data from established bioassays as well as to results from chemical analysis. Mechanism-specific biotests indicated a defined hazard potential of the sediment extracts, and microarray analysis revealed several classes of significantly regulated genes which could be related to the hazard potential. Results indicate that potential classes of contaminants can be assigned to sediment extracts by both classical biomarker genes and corresponding expression profile analyses of known substances. However, it is difficult to distinguish between specific responses and more universal detoxification of the organism.

  • 34.
    Legradi, J. B.
    et al.
    Institute for Environmental Research, Department of Ecosystem Analysis, ABBt–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany; Environment and Health, VU University, Amsterdam, Netherlands.
    Di Paolo, C.
    Institute for Environmental Research, Department of Ecosystem Analysis, ABBt–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Kraak, M. H. S.
    FAME-Freshwater and Marine Ecology, Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, Amsterdam, Netherlands.
    van der Geest, H. G.
    FAME-Freshwater and Marine Ecology, Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, Amsterdam, Netherlands.
    Schymanski, E. L.
    Luxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, Belvaux, Luxembourg.
    Williams, A. J.
    National Center for Computational Toxicology, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park NC, United States.
    Dingemans, M. M. L.
    KWR Watercycle Research Institute, Nieuwegein, Netherlands.
    Massei, R.
    Department Effect-Directed Analysis, Helmholtz Centre for Environmental Research-UFZ, Leipzig, Germany.
    Brack, W.
    Department Effect-Directed Analysis, Helmholtz Centre for Environmental Research-UFZ, Leipzig, Germany.
    Cousin, X.
    Ifremer, UMR MARBEC, Laboratoire Adaptation et Adaptabilités des Animaux et des Systèmes, Palavas-les-Flots, France; INRA, UMR GABI, INRA, AgroParisTech, Domaine de Vilvert, Jouy-en-Josas, France.
    Begout, M. -L
    Ifremer, Laboratoire Ressources Halieutiques, L’Houmeau, France.
    van der Oost, R.
    Department of Technology, Research and Engineering, Waternet Institute for the Urban Water Cycle, Amsterdam, Netherlands.
    Carion, A.
    Laboratory of Evolutionary and Adaptive Physiology, Institute of Life, Earth and Environment, University of Namur, Namur, Belgium.
    Suarez-Ulloa, V.
    Laboratory of Evolutionary and Adaptive Physiology, Institute of Life, Earth and Environment, University of Namur, Namur, Belgium.
    Silvestre, F.
    Laboratory of Evolutionary and Adaptive Physiology, Institute of Life, Earth and Environment, University of Namur, Namur, Belgium.
    Escher, B. I.
    Department of Cell Toxicology, Helmholtz Centre for Environmental Research-UFZ, Leipzig, Germany; Eberhard Karls University Tübingen, Environmental Toxicology, Center for Applied Geosciences, Tübingen, Germany.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Nilén, Greta
    Örebro University, School of Science and Technology.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Pollet, D.
    Faculty of Chemical Engineering and Biotechnology, University of Applied Sciences Darmstadt, Darmstadt, Germany.
    Waldmann, P.
    Faculty of Chemical Engineering and Biotechnology, University of Applied Sciences Darmstadt, Darmstadt, Germany.
    Kienle, C.
    Swiss Centre for Applied Ecotoxicology Eawag-EPFL, Dübendorf, Switzerland.
    Werner, I.
    Swiss Centre for Applied Ecotoxicology Eawag-EPFL, Dübendorf, Switzerland.
    Haigis, A. -C
    Institute for Environmental Research, Department of Ecosystem Analysis, ABBt–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Knapen, D.
    Zebrafishlab, Veterinary Physiology and Biochemistry, University of Antwerp, Wilrijk, Belgium.
    Vergauwen, L.
    Zebrafishlab, Veterinary Physiology and Biochemistry, University of Antwerp, Wilrijk, Belgium.
    Spehr, M.
    Institute for Biology II, Department of Chemosensation, RWTH Aachen University, Aachen, Germany.
    Schulz, W.
    Zweckverband Landeswasserversorgung, Langenau, Germany.
    Busch, W.
    Department of Bioanalytical Ecotoxicology, UFZ–Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Leuthold, D.
    Department of Bioanalytical Ecotoxicology, UFZ–Helmholtz Centre for Environmental Research, Leipzig, Germany.
    Scholz, S.
    Department of Bioanalytical Ecotoxicology, UFZ–Helmholtz Centre for Environmental Research, Leipzig, Germany.
    vom Berg, C. M.
    Department of Environmental Toxicology, Swiss Federal Institute of Aquatic Science and Technology, Eawag, Dübendorf, Switzerland.
    Basu, N.
    Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, Canada.
    Murphy, C. A.
    Department of Fisheries and Wildlife, Michigan State University, East Lansing, United States.
    Lampert, A.
    Institute of Physiology (Neurophysiology), Aachen, Germany.
    Kuckelkorn, J.
    Section Toxicology of Drinking Water and Swimming Pool Water, Federal Environment Agency (UBA), Bad Elster, Germany.
    Grummt, T.
    Section Toxicology of Drinking Water and Swimming Pool Water, Federal Environment Agency (UBA), Bad Elster, Germany.
    Hollert, H.
    Institute for Environmental Research, Department of Ecosystem Analysis, ABBt–Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    An ecotoxicological view on neurotoxicity assessment2018In: Environmental Sciences Europe, ISSN 2190-4707, E-ISSN 2190-4715, Vol. 30, article id 46Article, review/survey (Refereed)
    Abstract [en]

    The numbers of potential neurotoxicants in the environment are raising and pose a great risk for humans and the environment. Currently neurotoxicity assessment is mostly performed to predict and prevent harm to human populations. Despite all the efforts invested in the last years in developing novel in vitro or in silico test systems, in vivo tests with rodents are still the only accepted test for neurotoxicity risk assessment in Europe. Despite an increasing number of reports of species showing altered behaviour, neurotoxicity assessment for species in the environment is not required and therefore mostly not performed. Considering the increasing numbers of environmental contaminants with potential neurotoxic potential, eco-neurotoxicity should be also considered in risk assessment. In order to do so novel test systems are needed that can cope with species differences within ecosystems. In the field, online-biomonitoring systems using behavioural information could be used to detect neurotoxic effects and effect-directed analyses could be applied to identify the neurotoxicants causing the effect. Additionally, toxic pressure calculations in combination with mixture modelling could use environmental chemical monitoring data to predict adverse effects and prioritize pollutants for laboratory testing. Cheminformatics based on computational toxicological data from in vitro and in vivo studies could help to identify potential neurotoxicants. An array of in vitro assays covering different modes of action could be applied to screen compounds for neurotoxicity. The selection of in vitro assays could be guided by AOPs relevant for eco-neurotoxicity. In order to be able to perform risk assessment for eco-neurotoxicity, methods need to focus on the most sensitive species in an ecosystem. A test battery using species from different trophic levels might be the best approach. To implement eco-neurotoxicity assessment into European risk assessment, cheminformatics and in vitro screening tests could be used as first approach to identify eco-neurotoxic pollutants. In a second step, a small species test battery could be applied to assess the risks of ecosystems.

  • 35.
    Meyer-Alert, Henriette
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Ladermann, Kim
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Schiwy, Sabrina
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt – Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen H.
    Örebro University, School of Science and Technology.
    A temporal high-resolution investigation of the Ah-receptor pathway during early development of zebrafish (Danio rerio)2018In: Aquatic Toxicology, ISSN 0166-445X, E-ISSN 1879-1514, Vol. 204, p. 117-129Article in journal (Refereed)
    Abstract [en]

    In order to contribute to a comprehensive understanding of the regulating mechanisms of the aryl-hydrocarbon-receptor (AHR) in zebrafish embryos, we aimed to elucidate the interaction of proteins taking part in this signaling pathway during early development of the zebrafish (Danio rerio) after chemical exposure. We managed to illustrate initial transcription processes of the implemented proteins after exposure to two environmentally relevant chemicals: polychlorinated biphenyl 126 (PCB126) and β-Naphthoflavone (BNF). Using qPCR, we quantified mRNA every 4 h until 118 h post fertilization and found the expression of biotransformation enzymes (cyp1 family) and the repressor of the AHR (ahr-r) to be dependent on the duration of chemical exposure and the biodegradability of the compounds. PCB126 induced persistently increased amounts of transcripts as it is not metabolized, whereas activation by BNF was limited to the initial period of exposure. We did not find a clear relation between the amount of transcripts and activity of the induced CYP-proteins, so posttranscriptional mechanisms are likely to regulate biotransformation of BNF. With regard to zebrafish embryos and their application in risk assessment of hazardous chemicals, our examination of the AHR pathway especially supports the relevance of the time point or period of exposure that is used for bioanalytical investigations and consideration of chemical properties determining biodegradability.

  • 36.
    Meyer-Alert, Henriette
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Benzo[a]pyrene and 2,3-benzofuran induce divergent temporal patterns of AhR-regulated responses in zebrafish embryos (Danio rerio)2019In: Ecotoxicology and Environmental Safety, ISSN 0147-6513, E-ISSN 1090-2414, Vol. 183, article id UNSP 109505Article in journal (Refereed)
    Abstract [en]

    Biotests like the fish embryo toxicity test have become increasingly popular in risk assessment and evaluation of chemicals found in the environment. The large range of possible endpoints is a big advantage when researching on the mode of action of a certain substance. Here, we utilized the frequently used model organism zebrafish (Danio rerio) to examine regulative mechanisms in the pathway of the aryl-hydrocarbon receptor (AHR) in early development. We exposed embryos to representatives of two chemical classes known to elicit dioxin-like activity: benzo[a]pyrene for polycyclic aromatic hydrocarbons (PAHs) and 2,3-benzofuran for polar O-substituted heterocycles as a member of heterocyclic compounds in general (N-, S-, O-heterocycles; NSO-hets). We measured gene transcription of the induced P450 cytochromes (cyp1), their formation of protein and biotransformation activity throughout the whole embryonic development until 5 days after fertilization. The results show a very specific time course of transcription depending on the chemical properties (e.g. halogenation, planarity, Kow), the physical decay and the biodegradability of the tested compound. However, although this temporal pattern was not precisely transferable onto the protein level, significant regulation in enzymatic activity over time could be detected. We conclude, that a careful choice of time and end point as well as consideration of the chemical properties of a substance are fairly important when planning, conducting and especially evaluating biotests.

  • 37.
    O’Donovan, Sarit
    et al.
    Centre for Marine and Environmental Research, University of Algarve, Faro, Portugal.
    Mestre, Nélia C.
    Centre for Marine and Environmental Research, University of Algarve, Faro, Portugal.
    Abel, Serena
    Centre for Marine and Environmental Research, University of Algarve, Faro, Portugal.
    Fonseca, Tainá G.
    Centre for Marine and Environmental Research, University of Algarve, Faro, Portugal.
    Carteny, Camilla C.
    Systemic Physiological and Ecotoxicological Research, Department of Biology, University of Antwerp, Antwerp, Belgium.
    Cormier, Bettie
    Örebro University, School of Science and Technology. UMR CNRS 5805 EPOC, University of Bordeaux, Talence, France.
    Keiter, Steffen H.
    Örebro University, School of Science and Technology.
    Bebianno, Maria J.
    Centre for Marine and Environmental Research, University of Algarve, Faro, Portugal.
    Ecotoxicological Effects of Chemical Contaminants Adsorbed to Microplastics in the Clam Scrobicularia plana2018In: Frontiers in Marine Science, E-ISSN 2296-7745, Vol. 5, article id 143Article in journal (Refereed)
    Abstract [en]

    Although microplastics are distributed globally in the marine environment, a great deal of unknowns relating to their ecotoxicological effects on the marine biota remain. Due to their lipophilic nature, microplastics have the potential to adsorb persistent organic pollutants present in contaminated regions, which may increase their detrimental impact once assimilated by organisms. This study investigates the ecotoxicological effects of exposure to low-density polyethylene (LDPE) microplastics (11 - 13 µm), with and without adsorbed contaminants (benzo[a]pyrene - BaP and perfluorooctane sulfonic acid - PFOS), in the peppery furrow shell clam, Scrobicularia plana. Environmentally relevant concentrations of contaminants (BaP - 16.87±0.22 µg g-1 and PFOS - 70.22±12.41 µg g-1) were adsorbed to microplastics to evaluate the potential role of plastic particles as a source of chemical contamination once ingested. S. plana were exposed to microplastics, at a concentration of 1 mg L-1, in a water-sediment exposure setup for 14 days. Clams were sampled at the beginning of the experiment (day 0) and after 3, 7 and 14 days. BaP accumulation, in whole clam tissues, was analysed. A multi-biomarker assessment was conducted in the gills, digestive gland, and haemolymph of clams to clarify the effects of exposure. This included the quantification of antioxidant (superoxide dismutase, catalase, glutathione peroxidase) and biotransformation (glutathione-S-transferases) enzyme activities, oxidative damage (lipid peroxidation levels), genotoxicity (single and double strand DNA breaks), and neurotoxicity (acetylcholinesterase activity). Results suggest a potential mechanical injury of gills caused by ingestion of microplastics that may also affect the analysed biomarkers. The digestive gland seems less affected by mechanical damage caused by virgin microplastic exposure, with the MP-adsorbed BaP and PFOS exerting a negative influence over the assessed biomarkers in this tissue.

  • 38. Otte, Jens C.
    et al.
    Andersson, Carin
    Abrahamson, Alexandra
    Olsman Takner, Helena
    Örebro University, Department of Natural Sciences.
    Keiter, Steffen
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Hollert, Henner
    Brunström, Björn
    A bioassay approach to determine the dioxin-like activity in sediment extracts from the Danube River: ethoxyresorufin-O-deethylase induction in gill filaments and liver of three-spined sticklebacks (Gasterosteus aculeatus L.)2008In: Environment International, ISSN 0160-4120, E-ISSN 1873-6750, Vol. 34, no 8, p. 1176-1184Article in journal (Refereed)
    Abstract [en]

    Sediment samples from the upper Danube River in Germany have previously been characterized as ecotoxicologically hazardous and contaminants in these sediments may contribute to the observed decline of fish populations in this river section. For the investigation of sediment toxicity there is a need for development, standardization and implementation of in vivo test systems using vertebrates. Therefore, the main objective of this study was to apply and evaluate a recently established fish gill EROD assay as a biomarker in sediment toxicity assessment by using extracts of well characterised sediment samples from the upper Danube River. This to our knowledge is the first application of this novel assay to sediment extracts. Sediments from four different sites along the upper Danube River were Soxhlet-extracted with acetone and dissolved in DMSO. Three-spined sticklebacks (Gasterosteus aculeatus L.) were exposed for 48 h to various concentrations of the extracts, to the positive control β-naphthoflavone or to the solvent. Measurements of EROD activity in gill filaments and liver microsomes followed the exposure. Concentration-dependent induction of EROD in both gill and liver was found for all sediment extracts. The highest EROD-inducing potency was determined for extracts of sediments from the sites “Öpfinger See” and “Sigmaringen” and the EROD activities in gill and liver correlated well. The results from the gill and liver assays were in accordance with in vitro results of previous investigations. The EROD activities measured in the present study corresponded with the concentrations of PAHs, PCBs and PCDD/Fs in the sediment samples derived in a previous study. The sticklebacks in this study were in the reproductive phase and a stronger EROD induction was obtained in the females than in the males. Implementation of the EROD assay in testing of sediment extracts gave highly reliable results which make this assay an ecotoxicologically relevant method for assessment of contamination with Ah receptor agonists in sediments.

  • 39.
    Otte, Jens
    et al.
    Institute of Toxicology and Genetics, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.
    Keiter, Steffen
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Fassbender, Christopher
    Centre of Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Higley, Eric
    Toxicology Centre, University of Saskatchewan, Saskatoon, Canada.
    Suares Rocha, Paula
    Centre of Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Brinkmann, Markus
    Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Wahrendorf, Dierk-Steffen
    Department Bio-Chemistry and Ecotoxicology, Federal Institute of Hydrology, Koblenz, Germany.
    Manz, Werner
    Institut für Integrierte Naturwissenschaften, University of Koblenz-Landau, Koblenz, Germany.
    Wetzel, Markus
    Department of Animal Ecology, Federal Institute of Hydrology, Koblenz, Germany.
    Braunbeck, Thomas
    Centre of Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Giesy, John
    Toxicology Centre, University of Saskatchewan, Saskatoon, Canada; Department of Biomedical Veterinary Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Canada; Department of Zoology and Center for Integrative Toxicology, Michigan State University, East Lansing MI, United States of America; Department of Biology and Chemistry, and State Key Laboratory in Marine Pollution, City University of Hong Kong, Kowloon, China; School of Biological Sciences, University of Hong Kong, Hong Kong SAR, China.
    Hecker, Markus
    Toxicology Centre, University of Saskatchewan, Saskatoon, Canada; School of the Environment & Sustainability and Toxicology Centre, University of Saskatchewan, Saskatoon, Canada.
    Hollert, Henner
    RWTH Aachen University, Institute for Environmental Research, Aachen, Germany.
    Contribution of Priority PAHs and POPs to Ah Receptor-Mediated Activities in Sediment Samples from the RiverElbe Estuary, Germany2013In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 8, no 10, article id e7596Article in journal (Refereed)
    Abstract [en]

    The estuary of the River Elbe between Hamburg and the North Sea (Germany) is a sink for contaminated sediment and suspended particulate matter (SPM). One major concern is the effect of human activities on the hydrodynamics, particularlythe intensive dredging activities in this area that may result in remobilization of sediment-bound pollutants. The aim of this study was to identify pollutants contributing to the toxicological risk associated with re-suspension of sediments in the Elbe Estuary by use of an effect-directed analysis that combines chemical and biological analyses in with specific fractionation techniques. Sediments were collected from sites along the Elbe Estuary and a site from a small harbor basin of the Elbe Estuary that is known to be polluted. The sixteen priority EPA-PAHs were quantified in organic extracts of sediments. In addition, dioxin equivalents of sediments were investigated by use of the 7-ethoxyresorufin O-deethylase assay with RTL-W1 cells and the Ah receptor-mediated luciferase transactivation assay with H4IIE-luc cells. Quantification of the 16 priorityPAHs revealed that sediments were moderately contaminated at all of the sites in the Elbe River Estuary (,0.02–0.906 mg/gdw). Sediments contained relatively small concentrations of dioxin equivalents (Bio-TEQ) with concentrations ranging from15.5 to 322 pg/g dw, which were significantly correlated with dioxin equivalents calculated based on toxicity referencevalues and concentrations of PAH. The concentration of Bio-TEQ at the reference site exceeded 200,000 pg/g dw. In apotency balance the 16 PAHs explained between 47 and 118% of the Bio-TEQ in the luciferase assay, which can be explained by the constant input of PAHs bound to SPM from the upper course of the Elbe River into its estuary. Successful identification of a significant portion of dioxin-like activity to priority PAHs in complex environmental samples such assediments has rarely been reported.

  • 40.
    Peddinghaus, Sabrina
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Brinkmann, Markus
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Bluhm, Kerstin
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Sagner, Anne
    Department of Environmental Biotechnology, Water Technology Center (TZW), Karlsruhe, Germany.
    Hinger, Gunnar
    Aquatic Ecology and Toxicology, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Braunbeck, Thomas
    Aquatic Ecology and Toxicology, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
    Eisenträger, Adolf
    Dept. IV 2: Pharmaceuticals, Chemicals, Experimental Studies, Federal Environment Agency, Dessau, Germany; Institute of Hygiene and Environmental, Medicine, Medical Faculty, RWTH Aachen University, Aachen, Germany.
    Thiem, Andreas
    Department of Environmental Biotechnology, Water Technology Center (TZW), Karlsruhe, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen University, Aachen, Germany.
    Quantitative assessment of the embryotoxic potential of NSO-heterocyclic compounds using zebrafish (Danio rerio)2012In: Reproductive Toxicology, ISSN 0890-6238, E-ISSN 1873-1708, Vol. 33, p. 224-232Article in journal (Refereed)
    Abstract [en]

    Heterocyclic aromatic compounds (NSO-HET) have frequently been detected in the environment. Several studies have concluded that NSO-HET pose a threat to organisms in waters, sediments and soils. However, few publications are available assessing the ecotoxicology of NSO-HET. The present study aims toassess the embryo toxicity of heterocycles using Danio rerio. A combination of the Fish Embryo Toxicity Test and analytical quantification should aid to determine the hazard potential. Changes of the total concentrations due to sorption or volatility were quantified by GC/MS. Loss of compounds during the testwas observed primarily for volatile or hydrophobic NSO-HET. The LC50 calculated with nominal concentrations underestimates the toxicity by a factor up to 16 (2 h), demonstrating that a chemical analysis for comparing nominal and measured concentrations is essential for such investigations.

  • 41.
    Pittura, Lucia
    et al.
    Laboratorio di Ecotossicologia e Chimica ambientale, Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona, Italy.
    Avio, Carlo G.
    Laboratorio di Ecotossicologia e Chimica ambientale, Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona, Italy.
    Giuliani, Maria E.
    Laboratorio di Ecotossicologia e Chimica ambientale, Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona, Italy.
    d’Errico, Giuseppe
    Laboratorio di Ecotossicologia e Chimica ambientale, Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona, Italy.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Cormier, Bettie
    Örebro University, School of Science and Technology. UMR Centre National de la Recherche Scientifique 5805 EPOC, University of Bordeaux, Talence, France.
    Gorbi, Stefania
    Laboratorio di Ecotossicologia e Chimica ambientale, Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona, Italy.
    Regoli, Francesco
    Laboratorio di Ecotossicologia e Chimica ambientale, Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona, Italy; Consorzio Interuniversitario per le Scienze del Mare, CoNISMa, ULR Ancona, Ancona, Italy.
    Microplastics as Vehicles of Environmental PAHs to Marine Organisms: Combined Chemical and Physical Hazards to the Mediterranean Mussels, Mytilus galloprovincialis2018In: Frontiers in Marine Science, E-ISSN 2296-7745, Vol. 5, no Apr, article id 103Article in journal (Refereed)
    Abstract [en]

    The ubiquitous occurrence of microplastics (MPs) in the marine environment is raising concern for interactions with marine organisms. These particles efficiently adsorb persistent organic pollutants from surrounding environment and, due to the small size, they are easily available for ingestion at all trophic levels. Once ingested, MPs can induce mechanical damage, sub- lethal effects and various cellular responses, further modulated by possible release of adsorbed chemicals or additives. In this study, ecotoxicological effects of MPs and their interactions with benzo(a)pyrene (BaP), chosen as a model compound for polycyclic aromatic hydrocarbons (PAHs) were investigated in Mediterranean mussels, Mytilus galloprovincialis. Organisms were exposed for four weeks to 10 mg/L of low-density polyethylene (LD-PE) microparticles (2.34x107 particles/L, size range 20-25 µm), both virgin and pre-contaminated with BaP (15µg/g). Organisms were also exposed for comparison to BaP dosed alone at 150 ng/L, corresponding to the amount adsorbed on microplastics. Tissue localization of microplastics was histologically evaluated; chemical analyses and a wide battery of biomarkers covering molecular, biochemical and cellular levels allowed to evaluate BaP bioaccumulation, alterations of immune system, antioxidant defenses, onset of oxidative stress, peroxisomal proliferation, genotoxicity and neurotoxicity. Obtained data were elaborated within a quantitative weight of evidence (WOE) model which, using weighted criteria, provided synthetic hazard indices, for both chemical and cellular results, before their integration in a combined index. Microplastics were localized in haemolymph, gills and especially digestive tissues where a potential transfer of BaP from MPs was also observed. Significant alterations were measured on the immune system, while more limited effects occurred on the oxidative status, neurotoxicity and genotoxicity, with a different susceptibility of analyzed pathways, depending on tissue, time and typology of exposure. Molecular analyses confirmed the general lack of significant variations on transcriptional activity of antioxidant and stress genes. The overall results suggest that microplastics induce a slight cellular toxicity under short-term (28 days) exposure conditions. However, modulation of immune responses, along with bioaccumulation of BaP, pose the still unexplored risk that these particles, under conditions of more chronic exposure (months to years) or interacting with other stressors, may provoke long-term, subtle effects on organisms’ health status.

  • 42.
    Schiwy, Sabrina
    et al.
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen, Aachen, Germany.
    Braeunig, Jennifer
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen, Aachen, Germany.
    Alert, Henriette
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen, Aachen, Germany.
    Hollert, Henner
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen, Aachen, Germany.
    Keiter, Steffen
    Department of Ecosystem Analysis, Institute for Environmental Research, RWTH Aachen, Aachen, Germany.
    A novel contact assay for testing aryl hydrocarbon receptor (AhR): mediated toxicity of chemicals and whole sediments in zebrafish (Danio rerio) embryos2015In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 22, no 21, p. 16305-16318Article in journal (Refereed)
    Abstract [en]

    The European Water Framework Directive aims to achieve a good ecological and chemical status in surface waters until 2015. Sediment toxicology plays a major role in this intention as sediments can act as a secondary source of pollution. In order to fulfill this legal obligation, there is an urgent need to develop whole-sediment exposure protocols, since sediment contact assays represent the most realistic scenario to simulate in situ exposure conditions. Therefore, in the present study, a vertebrate sediment contact assay to determine aryl hydrocarbon receptor (AhR)-mediated activity of particle-bound pollutants was developed. Furthermore, the activity and the expression of the CYP1 family in early lifestages of zebrafish after exposure to freeze-dried sediment samples were investigated. In order to validate the developed protocol, effects of β-naphthoflavone and three selected sedimenton zebrafish embryos were investigated. Results documented clearly AhR-mediated toxicity after exposure to β-naphthoflavone (β-NF) and to the sediment from the Vering canal. Upregulation of mRNA levels was observed for all investigated sediment samples. The highest levels of all investigated cyp genes (cyp1a, cyp1b1, cyp1c1, and cyp1c2) were recorded after exposure to the sediment sample of the Vering canal. In conclusion, the newly developed sediment contact assay can be recommended for the investigation of dioxin-like activities of single substances and the bioavailable fraction of complex environmental samples. Moreover, the exposure of whole zebrafish embryos to native (freeze-dried) sediment samples represents a highly realistic and ecologically relevant exposure scenario.

  • 43.
    Seitz, Nadja
    et al.
    University of Heidelberg, Heidelberg, Germany.
    Böttcher, Melanie
    University of Heidelberg, Heidelberg, Germany.
    Keiter, Steffen
    University of Heidelberg, Heidelberg, Germany.
    Kosmehl, Thomas
    University of Heidelberg, Heidelberg, Germany.
    Manz, Werner
    Federal Institute of Hydrology, Koblenz, Germany.
    Hollert, Henner
    University of Heidelberg, Heidelberg, Germany; Aachen University, Aachen, Germany.
    Braunbeck, Thomas
    University of Heidelberg, Heidelberg, Germany.
    A novel statistical approach for the evaluation of comet assay data2008In: Mutation Research, ISSN 1383-5742, E-ISSN 1388-2139, Vol. 652, no 1, p. 38-45Article in journal (Refereed)
    Abstract [en]

    The present study forms part of a weight-of-evidence framework including genotoxicological studies in the upper Danube River basin, which aim at elucidating the reasons for the decline in fish catch. The major focus of this paper is the assessment of genotoxicity of sediments from the Danube River basin by use of the comet assay with RTL-W1 cells and with embryos of zebrafish (Danio rerio). A frequently discussed question in this type of approach is how to aggregate and compare the data obtained from genotoxicity testing. There is a need to develop mathematical method combining the information from dose–response curves and level of effectiveness (maximum genotoxic effect). For comparison and ranking of the genotoxic potential of samples from different locations along the Danube River, several methods based on EC50, Lowest Observed Effect Concentration (LOEC), and maximum induction factor were compared with respect to their validity. An evaluation system termed the “3-step, analysis” was developed to facilitate consideration of a maximum number of aspects of the raw data. The so-called “concentration-dependent induction factor” (CDI) introduces an index for a straightforward, precise and realistic assessment of the genotoxic potential of any kind of field sample or genotoxic agent.

  • 44.
    Sjöberg, Viktor
    et al.
    Örebro University, School of Science and Technology.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Karlsson, Stefan
    Örebro University, School of Science and Technology.
    Metal toxicity: Are we measuring what we want?2015Conference paper (Other academic)
  • 45.
    Suares Rocha, Paula
    et al.
    University of Heidelberg.
    Keiter, Steffen
    Örebro University, School of Science and Technology.
    Pompeo, Marcelo Luiz Martins
    University of Sao Paulo.
    Mariani, Carolina Fioriollo
    University of Sao Paulo.
    Brandimarte, Ana Lúcia
    University of Sao Paulo.
    Seiler, Thomas-Benjamin
    University of Heidelberg.
    Kosmehl, Thomas
    University of Heidelberg.
    Böttcher, Melanie
    University of Heidelberg.
    Wölz, Jan
    University of Heidelberg.
    Braunbeck, Thomas
    University of Heidelberg.
    Storch, Volker
    University of Heidelberg.
    Hollert, Henner
    University of Heidelberg.
    Weight-of-Evidence-Studie zur Sedimentbelastung des Tietê River in Brasilien2006In: Umweltwissenschaften und Schadstoff-Forschung, ISSN 0934-3504, Vol. 18, no 1, p. 70-Article in journal (Other academic)
1 - 45 of 45
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