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  • 1.
    Andersson, Erika
    et al.
    Örebro University, School of Science and Technology.
    Rotander, Anna
    Örebro University, School of Science and Technology.
    von Kronhelm, Thomas
    SAKAB.
    Berggren, Anna
    Analycen AB.
    Ivarsson, Per
    Analycen AB.
    Hollert, Henner
    RWTH Aachen university.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    AhR agonist and genotoxicant bioavailability in a PAH-contaminated soil undergoing biological treatment2009In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 16, no 5, p. 521-530Article in journal (Refereed)
    Abstract [en]

    Degradation of the 16 US EPA priority PAHs in soil subjected to bioremediation is often achieved. However, the PAH loss is not always followed by a reduction in soil toxicity. For instance, bioanalytical testing of such soil using the chemical-activated luciferase gene expression (CALUX) assay, measuring the combined effect of all Ah receptor (AhR) activating compounds, occasionally indicates that the loss of PAHs does not correlate with the loss of Ah receptor-active compounds in the soil. In addition, standard PAH analysis does not address the issue of total toxicant bioavailability in bioremediated soil.

    To address these questions, we have used the CALUX AhR agonist bioassay and the Comet genotoxicity bioassay with RTL-W1 cells to evaluate the toxic potential of different extracts from a PAH-contaminated soil undergoing large-scale bioremediation. The extracts were also chemically analyzed for PAH16 and PCDD/PCDF. Soil sampled on five occasions between day 0 and day 274 of biological treatment was shaken with n-butanol with vortex mixing at room temperature to determine the bioavailable fraction of contaminants. To establish total concentrations, parts of the same samples were extracted using an accelerated solvent extractor (ASE) with toluene at 100A degrees C. The extracts were tested as inducers of AhR-dependent luciferase activity in the CALUX assay and for DNA breakage potential in the Comet bioassay.

    The chemical analysis of the toluene extracts indicated slow degradation rates and the CALUX assay indicated high levels of AhR agonists in the same extracts. Compared to day 0, the bioavailable fractions showed no decrease in AhR agonist activity during the treatment but rather an up-going trend, which was supported by increasing levels of PAHs and an increased effect in the Comet bioassay after 274 days. The bio-TEQs calculated using the CALUX assay were higher than the TEQs calculated from chemical analysis in both extracts, indicating that there are additional toxic PAHs in both extracts that are not included in the chemically derived TEQ.

    The response in the CALUX and the Comet bioassays as well as the chemical analysis indicate that the soil might be more toxic to organisms living in soil after 274 days of treatment than in the untreated soil, due to the release of previously sorbed PAHs and possibly also metabolic formation of novel toxicants.

    Our results put focus on the issue of slow degradation rates and bioavailability of PAHs during large-scale bioremediation treatments. The release of sorbed PAHs at the investigated PAH-contaminated site seemed to be faster than the degradation rate, which demonstrates the importance of considering the bioavailable fraction of contaminants during a bioremediation process.

    It has to be ensured that soft remediation methods like biodegradation or the natural remediation approach do not result in the mobilization of toxic compounds including more mobile degradation products. For PAH-contaminated sites this cannot be assured merely by monitoring the 16 target PAHs. The combined use of a battery of biotests for different types of PAH effects such as the CALUX and the Comet assay together with bioavailability extraction methods may be a useful screening tool of bioremediation processes of PAH-contaminated soil and contribute to a more accurate risk assessment. If the bioremediation causes a release of bound PAHs that are left undegraded in an easily extracted fraction, the soil may be more toxic to organisms living in the soil as a result of the treatment. A prolonged treatment time may be one way to reduce the risk of remaining mobile PAHs. In critical cases, the remediation concept might have to be changed to ex situ remediation methods.

  • 2. Bergknut, Magnus
    et al.
    Kucera, Adam
    Frech, Kristina
    Andersson, Erika
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Rannug, Ulf
    Koci, Vladimir
    Andersson, Patrik L.
    Haglund, Peter
    Tysklind, Mats
    Identification of potentially toxic compounds in complex extracts of environmental samples using gas chromatography-mass spectrometry and multivariate data analysis2007In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 26, no 2, p. 208-217Article in journal (Refereed)
    Abstract [en]

    In this study, we examined 31 samples of varying chemical composition, including samples of soils from gasworks, coke production sites, and sites where wood preservatives were heavily used; ash and soot from municipal solid waste incinerators; antiskid sand; and dust from areas with heavy road traffic. The samples were comprehensively chemically characterized, especially their polycyclic aromatic compound contents, using gas chromatography-time-of-flight mass spectrometry, whereas their biological effects were assessed using dehydrogenase activity, root growth (Hordeum vulgare), reproduction of springtails (Folsomia candida), algal growth (Desmodesmus subspicatus), germinability (Sinapis alba), Vibrio fischeri, DR-CALUX, and Ames Salmonella assays. The number of compounds detected in the samples ranged from 123 to 527. Using the multivariate regression technique of partial-least-squares projections to latent structures, it was possible to find individual compounds that exhibited strong correlations with the different biological responses. Some of the results, however, indicate that a broader chemical characterization may be needed to identify all the compounds that may cause the measured biological responses.

  • 3.
    Brunström, Björn
    et al.
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Engwall, Magnus
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Hjelm, Katarina
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Lindqvist, Lars
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Zebuhr, Yngve
    Aquatic Chemical Ecotoxicology, Department of Zoology, University of Stockholm, Stockholm, Sweden.
    EROD induction in cultured chick embryo liver: a sensitive bioassay for dioxin-like environmental pollutants1995In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 14, no 5, p. 837-842Article in journal (Refereed)
    Abstract [en]

    A technique for studying 7-ethoxyresorufin O-deethylase (EROD) induction in chick embryo liver in vitro was developed. Livers from 8-d-old embryos were cultured in rotating vials at 37 degrees C for 48 h in a medium to which DMSO-dissolved test compounds had been added. This bioassay proved to be highly sensitive to dioxin-like compounds, and its usefulness for assessing the toxic potency of such compounds in environmental samples was demonstrated. Concentration-response curves were determined for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 3,3',4,4',5-penlachlorobiphenyl (PCB IUPAC no. 126), 3,3',4,4'-tetrachlorobiphenyl (PCB 77), 2,3,3',4,4'-pentachlorobiphenyl (PCB 105), and benzo[k]fluoranthene (BkF). TCDD induced EROD in a concentration-dependent manner, having an EC50 of 5.0 x 10(-12) M. The cultured embryo livers were extremely sensitive to TCDD, and about 30 fg of this compound per liver (2 x 10(-13) M) was enough to significantly induce EROD. The EC50 values obtained for PCBs 126, 77, 105, and BkF were 4,4 x 10(-11) M, 9.2 x 10(-9) M, 1.6 x 10(-8) M, and 6.2 x 10(-7) M, respectively. The maximal induction levels obtained for three different preparations of polychlorinated naphthalenes (PCNs) were less than those of the other compounds tested. When the technical PCN mixture Halowax 1014 was coadministered with TCDD, the induction was lower than that caused by TCDD alone. An organic extract of fly ash from a municipal waste combustion plant was very potent. Considering its contents of polychlorinated dibenzo-p-dioxins/furans, expressed as TCDD equivalents, the EC50 obtained was close to that for TCDD.

  • 4.
    Buitrago, B. Huerta
    et al.
    Mälardalen University, Västerås, Sweden.
    Munoz, P. Ferrer
    Mälardalen University, Västerås, Sweden.
    Ribe, V.
    Mälardalen University, Västerås, Sweden.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Wojciechowska, E.
    Gdansk University of Technology, Gdansk, Poland.
    Waara, S.
    Mälardalen University, Västerås, Sweden.
    Hazard assessment of sediments from a wetland system for treatment of landfill leachate using bioassays2013In: Ecotoxicology and Environmental Safety, ISSN 0147-6513, E-ISSN 1090-2414, Vol. 97, p. 255-262Article in journal (Refereed)
    Abstract [en]

    Four bioassays were used in this study for the hazard assessment of sediments from sediment traps and several ponds in a treatment wetland for landfill leachate at Atleverket, Sweden. In the 6-day solid phase microbiotest with the sediment-dwelling crustacean Heterocypris incongruens both acute and chronic effects were observed with a gradual decrease and loss of toxicity with treatment in the wetland system. Some samples showed a low toxicity in porewater and only one sample was weakly toxic in the whole sediment test when assessed with Aliivibrio fischeri (Vibro fischeri). No genotoxicity was detected in the umu test. The toxicity response in the H4IIE- luc test evaluating the presence of dioxin-like compounds was considerably higher in the samples from the sediment traps. The hazard of the sediment therefore appears to be highest in the sediment traps and pond 1 with the methods employed. The result indicates that the wetland system has a design supporting the concentration and sequestration of toxic substances in the first part of the wetland. Based upon the results we suggest that hazard assessment of sediments from other treatment wetlands for landfill leachate should be conducted. (c) 2013 Elsevier Inc. All rights reserved.

  • 5.
    Eichbaum, Kathrin
    et al.
    RWTH Aachen University, Aachen, Germany.
    Brinkmann, Markus
    RWTH Aachen University, Aachen, Germany.
    Buchinger, Sebastian
    German Federal Institute of Hydrology (BfG), Koblenz, Germany.
    Hecker, Marcus
    University of Saskatchewan, Saskatoon, Canada.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    van Bavel, Bert
    Örebro University, School of Science and Technology.
    Reifferscheid, Georg
    German Federal Institute of Hydrology (BfG), Koblenz, Germany.
    Hollert, Henner
    RWTH Aachen University, Aachen, Germany.
    The dioRAMA project: assessment of dioxin-like activity in sediments and fish (Rutilus rutilus) in support of the ecotoxicological characterization of sediments2013In: Journal of Soils and Sediments, ISSN 1439-0108, E-ISSN 1614-7480, Vol. 13, no 4, p. 770-774Article in journal (Refereed)
    Abstract [en]

    Introduction: Given the complex interactions of re-suspension processes and bioavailability of sediment-bound pollutants such as dioxin-like chemicals, there is need for a better integrative understanding of the cause-effect relationship of these pollutants. Currently, the majority of studies investigating potential risks of these chemicals only focus on characterizing sediment extracts via in vitro bioassays, thereby disregarding bioavailability, uptake, metabolism, and elimination rates of these compounds in vivo. To determine to which extent mechanism-specific effects in vitro reflect possible adverse effects in vivo, the research project dioRAMA, involving partnership between the Institute for Environmental Research of RWTH Aachen University and the Department Biochemistry/Ecotoxicology of the German Federal Institute of Hydrology, was established.

    Methods: Animals from an elevated trophic level-common roach (Rutilus rutilus)-will be exposed to sediments from two major German rivers. Exposure will be performed in a system that enables a concurrent monitoring of environmental parameters. In parallel, in vitro studies will be conducted to determine dioxin-like potentials of sediment and fish extracts from the in vivo exposure experiments using different cell lines with varying endpoints. Moreover, extract fractionation procedures, using the strategy of effect-directed analysis, will enable the detection of specific contaminant groups responsible for the biological activity observed.

    Conclusion: A closer interconnection between applied ecotoxicological science and regulatory needs will facilitate the improved assessment of dioxin-like compounds in sediment and biota. Consequently, this will enable their application in sediment management programs, which is one of the main goals of the dioRAMA project.

  • 6.
    Eichbaum, Kathrin
    et al.
    Rhein Westfal TH Aachen, Dept Ecosystem Anal, Inst Environm Res, Aachen, Germany.
    Brinkmann, Markus
    Rhein Westfal TH Aachen, Dept Ecosystem Anal, Inst Environm Res, Aachen, Germany.
    Buchinger, Sebastian
    Fed Inst Hydrol BFG, Dept Biochem G3, Koblenz, Germany.
    Reifferscheid, Georg
    Fed Inst Hydrol BFG, Dept Biochem G3, Koblenz, Germany.
    Hecker, Markus
    Univ Saskatchewan, Sch Environm & Sustainabil, Saskatoon, Canada; Univ Saskatchewan, Toxicol Ctr, Saskatoon, Canada.
    Giesy, John P.
    Univ Saskatchewan, Sch Environm & Sustainabil, Saskatoon, Canada; Univ Saskatchewan, Toxicol Ctr, Saskatoon, Canada; Univ Saskatchewan, Dept Vet Biomed Sci, Saskatoon, Canada; Univ Saskatchewan, Toxicol Ctr, Saskatoon, Canada; Michigan State Univ, Dept Zool, E Lansing, USA; Michigan State Univ, Ctr Integrat Toxicol, E Lansing, USA; City Univ Hong Kong, State Key Lab Marine Pollut, Dept Biol & Chem, Kowloon, Hong Kong, Peoples R China; Univ Hong Kong, Sch Biol Sci, Hong Kong, Hong Kong, Peoples R China.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    van Bavel, Bert
    Örebro University, School of Science and Technology.
    Hollert, Henner
    Rhein Westfal TH Aachen, Dept Ecosystem Anal, Inst Environm Res, Aachen, Germany; Tongji Univ, Coll Environm Sci & Engn, Minist China, Key Lab Yangtze River Environm Educ, Shanghai 200092, Peoples R China; Chongqing Univ, Coll Resources & Environm Sci, Chongqing 400030, Peoples R China; Nanjing Univ, Sch Environm, Nanjing, Jiangsu, Peoples R China.
    In vitro bioassays for detecting dioxin-like activity: Application potentials and limits of detection, a review2014In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 487, p. 37-48Article in journal (Refereed)
    Abstract [en]

    Use of in vitro assays as screening tool to characterize contamination of a variety of environmental matrices has become an increasingly popular and powerful toolbox in the field of environmental toxicology.

    While bioassays cannot entirely substitute analytical methods such as gas chromatography-mass spectrometry (GC-MS), the increasing improvement of cell lines and standardization of bioassay procedures enhance their utility as bioanalytical pre-screening tests prior to more targeted chemical analytical investigations. Dioxin-receptor-based assays provide a holistic characterization of exposure to dioxin-like compounds (DLCs) by integrating their overall toxic potential, including potentials of unknown DLCs not detectable via e.g. GC-MS. Hence, they provide important additional information with respect to environmental risk assessment of DLCs.

    This review summarizes different in vitro bioassay applications for detection of DLCs and considers the comparability of bioassay and chemical analytically derived toxicity equivalents (TEQs) of different approaches and various matrices. These range from complex samples such as sediments through single reference to compound mixtures. A summary of bioassay derived detection limits (LODs) showed a number of current bioassays to be equally sensitive as chemical methodologies, but moreover revealed that most of the bioanalytical studies conducted to date did not report their LODs, which represents a limitation with regard to low potency samples.

    (C) 2014 Elsevier B.V. All rights reserved.

  • 7.
    Engwall, Magnus
    et al.
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Broman, Dag
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden; Inst. of Appl. Environ. Research, Laboratory for Aquatic Ecotoxicology, Stockholm University, Nyköping, Sweden .
    Dencker, Lennart
    Department of Toxicology, Uppsala University, Uppsala, Sweden.
    Näf, Carina
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden .
    Zebuhr, Yngve
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden .
    Brunström, Björn
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Toxic potencies of extracts of sediment and settling particulate matter collected in the recipient of a bleached pulp mill effluent before and after abandoning chlorine bleaching1997In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 16, no 6, p. 1187-1194Article in journal (Refereed)
    Abstract [en]

    Extracts of bottom sediment and settling particulate matter (SPM) samples collected in the recipient water body of a bleached pulp mill effluent were separated into three fractions: monoaromatic/aliphatic compounds, diaromatic compounds (e.g., polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans), and polyaromatic compounds assessment of dioxinlike potency were an in vitro assay, based on the induction of 7-ethoxyresorufin-O-deethylase (EROD) in chicken embryo livers, a test for EROD induction in ovo using chicken embryos, and an immunotoxicity test measuring inhibition of lymphoid cell development in cultured fetal mouse thymuses. The samples collected closest to the mill were the most potent. There was a time-dependent decrease in dioxinlike potency in the SPM samples collected near the mill, which coincided with the cessation of chlorine gas bleaching at the mill. The bioassays in general, and the in vitro chicken embryo liver bioassay in particular, proved useful in the assessment of dioxinlike compounds in the sediment and SPM samples. The polyaromatic fractions of the samples were generally more potent than the diaromatic fractions. Only a small part of the effects caused by the polyaromatic fractions could be explained by 15 analyzed PAHs, indicating the presence of unquantified polyaromatic compounds with dioxinlike effects. This investigation indicates that the cessation of chlorine bleaching in the pulp mill resulted in a reduced load of diaromatic dioxinlike compounds in the recipient water body.

  • 8.
    Engwall, Magnus
    et al.
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Broman, Dag
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden; Inst. of Appl. Environ. Research, Laboratory for Aquatic Ecotoxicology, Stockholm University, Nyköping, Sweden .
    Ishaq, Rasha
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden.
    Näf, Carina
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden.
    Zebuhr, Yngve
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden.
    Brunström, Björn
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Toxic potencies of lipophilic extracts from sediments and settling particulate matter (SPM) collected in a PCB-contaminated river system1996In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 15, no 2, p. 213-222Article in journal (Refereed)
    Abstract [en]

    Sediments and settling particulate matter (SPM) were sampled in a polychlorinated biphenyl (PCB)-contaminated lake, Lake Jarnsjon, and in lakes located up-/and downstream from Lake Jarnsjon. The 7-ethoxyresorufin O-deethylase (EROD)-inducing potencies of lipophilic extracts from sediment and SPM were investigated in organ cultures of chicken embryo livers (denoted in vitro chicken embryo liver bioassay). The extract from sediments collected in Lake Jarnsjon had the greatest EROD-inducing potency of the extracts studied, and it also contained the highest concentrations of PCBs, polycyclic aromatic hydrocarbons (PAHs), and polychlorinated dibenzodioxins/furans (PCDDs/Fs). The non-ortho-chlorinated PCBs in the extracts were probably major contributors to the EROD induction noted. The EROD-inducing potency of sediment extract from the lake downstream Lake Jarnsjon was higher than that of extract from the upstream lake. This indicates that EROD-inducing substances were transported from Lake Jarnsjon sediment to the lake downstream. The extracts from Lake Jarnsjon sediment and SPM were separated into three fractions containing aliphatic/monoaromatic, diaromatic, and polyaromatic compounds, respectively, which were tested in the in vitro chicken embryo Liver bioassay. In all extracts, the aliphatic/monoaromatic fractions were low in EROD-inducing potency, while the polyaromatic fractions from SPM were more potent than their corresponding diaromatic fractions, indicating that they contained high concentrations of aromatic hydrocarbon (Ah) receptor ligands. Only a small part of the EROD induction caused by the polyaromatic fractions could be attributed to the 15 analyzed PAHs in the samples. The EROD-inducing potencies of the diaromatic fractions correlated quite well with their concentrations of PCDDs/Fs and PCBs. The in vitro chicken embryo liver bioassay detected low concentrations of EROD inducers in the extracts and therefore proved useful for estimating the contents of Ah receptor ligands in environmental samples.

  • 9.
    Engwall, Magnus
    et al.
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Broman, Dag
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden; Inst. of Appl. Environ. Research, Laboratory for Aquatic Ecotoxicology, Stockholm University, Nyköping, Sweden .
    Näf, Carina
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden .
    Zebuhr, Yngve
    Aquatic Chemical Ecotoxicology, Department of Zoology, Stockholm University, Stockholm, Sweden .
    Brunström, Björn
    Dept. of Environmental Toxicology, Uppsala University, Uppsala, Sweden .
    Dioxin-like compounds in HPLC-fractionated extracts of marine samples from the east and west coast of Sweden: bioassay- and instrumentally-derived TCDD equivalents1997In: Marine Pollution Bulletin, ISSN 0025-326X, E-ISSN 1879-3363, Vol. 34, no 12, p. 1032-1040Article in journal (Refereed)
    Abstract [en]

    Lipophilic extracts of sediment, settling particulate matter (SPM) and blue mussel (Mytilus edulis) samples, collected at coastal locations on the east and west coast of Sweden, were HPLC-separated into three fractions containing 1. monoaromatic/aliphatic, 2. diaromatic (e,g, polychlorinated biphenyls (PCBs), polychlorinated dibenzodioxins and polychlorinated dibenzofuraus (PCDDs/Fs)), and 3, polyaromatic compounds (e,g, polycyclic aromatic hydrocarbons (PAHs)), The fractions were tested for dioxin-like effects using a sensitive bioassay, based on EROD-induction in cultured chicken embryo livers, The concentrations of PCDDs/Fs, PCBs and 15 PAHs in the samples were also determined, The polyaromatic fractions of the sediment samples were more potent as EROD-inducers than the diaromatic fractions, Only a small part of the EROD-induction caused by the polyaromatic fractions could be explained by the analysed PAHs in the samples, indicating presence of non-quantified polyaromatic EROD-inducing compounds in the samples, A greater pollution by EROD-inducing diaromatic and polyaromatic compounds on the east coast site than on the west coast site was seen, The filtration activities and faeces production of the mussels on the east coast experimental site increased the sedimentation of PCDDs/Fs, PCBs and PAHs, The described bioassay proved very useful in the assessment of dioxin-like compounds in both abiotic and biotic samples from the marine environment,

  • 10.
    Engwall, Magnus
    et al.
    Department of Environmental Toxicology, Uppsala University, Uppsala, Sweden.
    Brunström, Björn
    Department of Environmental Toxicology, Uppsala University, Uppsala, Sweden.
    Brewer, Andrea
    Department of Environmental Toxicology, Uppsala University, Uppsala, Sweden.
    Norrgren, Leif
    Department of Pathology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Cytochrome P450IA induction by a coplanar PCB, a PAH mixture, and PCB-contaminated sediment extracts following microinjection of rainbow trout sac-fry1994In: Aquatic Toxicology, ISSN 0166-445X, E-ISSN 1879-1514, Vol. 30, no 4, p. 311-324Article in journal (Refereed)
    Abstract [en]

    The 7-ethoxyresorufin-O-deethylase (EROD)-inducing potencies of a coplanar polychlorinated biphenyl (PCB) (3,3',4,3',5-pentachlorobiphenyl, IUPAC No. 126), a mixture of five polycyclic aromatic hydrocarbons (benzo(k)fluoranthene, benz(a)anthracene, benzo(a)pyrene, indeno(1,2,3-cd)pyrene and chrysene), and lipophilic compounds extracted from the sediments in a PCB-contaminated lake and from sediments in lakes up- and downstream, were studied in rainbow trout sac-fry in a 43-day study. The compounds/extracts were injected into the yolk sacs of newly hatched sac-fry and hepatic EROD activities and mortality rates were measured at various times after the injections. Five livers from each group were also examined by transmission electron microscopy. All the compounds/extracts induced hepatic EROD activities in the sac-fry. Ten days after injection the EROD activity caused by PCB No. 126 (1.3 ng per embryo) was 40-fold compared to the control activity. This was the highest induction rate observed in the experiment. For the sediment extracts, the highest induction rates were observed at the first sampling occasion, which for these groups was on day 24. The extract from the Lake Jarnsjon sediment was more potent as an EROD inducer than the extracts of sediments from the lakes up- and downstream from Lake Jarnsjon. None of the sediment extracts caused any significant mortality. In sac-fry injected with the PAH mixture, EROD was only slightly induced. The highest dose of PAHs (10 mu g per embryo) caused about 90% mortality by 24 days after injection. When the livers were examined by transmission electron microscopy, morphological alterations (e.g. hepatocyte degeneration and hypertrophy) were seen in the groups injected with Lake Jarnsjon sediment extract, PCB No. 126 and the highest dose of the PAH mixture (10 mu g per embryo).

  • 11.
    Engwall, Magnus
    et al.
    Section of Environmental Toxicology, Uppsala University, Uppsala, Sweden.
    Brunström, Björn
    Section of Environmental Toxicology, Uppsala University, Uppsala, Sweden.
    Jakobsson, Eva
    Environmental Chemistry, Wallenberg Laboratory, Stockholm University, Stockholm, Sweden.
    Ethoxyresorufin O-deethylase (EROD) and aryl hydrocarbon hydroxylase (AHH)-inducing potency and lethality of chlorinated naphthalenes in chicken (Gallus domesticus) and eider duck (Somateria mollissima) embryos1994In: Archives of Toxicology, ISSN 0340-5761, E-ISSN 1432-0738, Vol. 68, no 1, p. 37-42Article in journal (Refereed)
    Abstract [en]

    The 7-ethoxyresorufin O-deethylase (EROD)- and aryl hydrocarbon hydroxylase (AHH)-inducing potencies and lethalities of a technical preparation of polychlorinated naphthalenes (PCNs) (Halowax 1014, approx imate congener ratio: 20% tetrachloronaphthalenes, 40% pentachloronaphthalenes, 40% hexachloronaphthalenes), a mixture of 50% 1,2,3,5,6,7-hexachloronaphthalene and 50% 1,2,3,4,6,7-hexachloronaphthalene (HxCN-mix), and 1,2,3,4,5,6,7-heptachloronaphthalene (HpCN) were studied in chicken (Gallus domesticus) and elder duck (Somateria mollissima) embryos. Mortality and hepatic EROD activity were determined on day 10 of incubation in chicken embryos exposed to various doses of the PCNs via the air-sacs of the eggs on day 7. The HxCN-mix and Halowax 1014 proved to have both embryolethal and EROD-inducing properties, while the HpCN had low EROD-inducing potency and embryolethality. ED(50) values for EROD induction by the HxCN-mix and Halowax 1014 were estimated to be 0.06 mg/kg egg and 0.2 mg/kg egg, respectively. Fifty percent of the chicken embryos died (6/12) when given 3.0 mg/kg of the HxCN-mix while a similar dose of Halowax 1014 caused mortality in 4 out of 12 chicken embryos. The dose-response curve for EROD induction by Halowax 1014 exhibited a decline after the maximal level was reached. When Halowax 1014 (1.0 mg/kg, egg) was coinjected with 3,3',4,4',5-pentachlorobiphenyl (PCB IUPAC #126) (0.1 mu g/kg egg) no additive effects on EROD activity were found, but when the same dose of Halowax 1014 was coinjected with a dose of PCB #126, known to cause maximal induction (1.0 mu g/kg egg), the resulting EROD activity was lower than that caused solely by 1.0 mu g PCB #126/kg egg. These findings indicate that Halowax 1014 has both EROD-inducing and EROD-inhibiting properties. Mortality and EROD and AHH activities were determined on day 18 (chicken) or day 24 (elder) of incubation in embryos exposed to 1.0 mg/kg egg via the yolk-sac on day 4 (chicken) or day 5 (elder). The HxCN-mix and Halowax 1014 induced AHH and EROD in both chicken and elder, but the induction rates were higher in the elder embryos. The HxCN-mix and Halowax 1014 caused degenerative hepatic lesions and pericardial oedema in the chicken embryos but not in the elder embryos. The most toxic PCNs tested (the HxCN-mix and Halowax 1014) were approximately of the same EROD-inducing potency as previously found for the most toxic mono-ortho-chlorinated biphenyls (Brunstrom 1990), and 1000 times less toxic and potent as EROD inducers compared with PCB #126 (Brunstrom and Andersson 1988). HpCN was considerably less toxic and exhibited a low EROD-inducing potency. The chicken embryos were more sensitive to the hepatotoxic effects produced by Halowax 1014 and the HxCN-mix than the elder duck embryos, while the elder embryos were more responsive in terms of EROD and AHH induction. The two HxCNs studied usually make up approximately 1% of the total quantity of PCNs present in Halowax 1014 [when determined with gas chromatography (flame ionization detection)]. Therefore, the relatively high toxic potency of Halowax 1014 cannot be explained by its content of the two HxCNs.

  • 12.
    Engwall, Magnus
    et al.
    Dept. of Environmental Toxicology, Uppsala university, Uppsala, Sweden .
    Brunström, Björn
    Dept. of Environmental Toxicology, Uppsala university, Uppsala, Sweden .
    Näf, Carina
    Inst. of Appl. Environ. Res. - ITM, Stockholm University, Stockholm, Sweden.
    Hjelm, Katarina
    Dept. of Environmental Toxicology, Uppsala university, Uppsala, Sweden .
    Levels of dioxin-like compounds in sewage sludge determined with a bioassay based on EROD induction in chicken embryo liver cultures1999In: Chemosphere, ISSN 0045-6535, E-ISSN 1879-1298, Vol. 38, no 10, p. 2327-2343Article in journal (Refereed)
    Abstract [en]

    A bioassay for the detection of dioxin-like compounds was used to estimate levels in sewage sludge from Swedish sewage treatment plants (STPs). The sludge extracts were HPLC-separated into three fractions containing a) monoaromatic/aliphatic, b) diaromatic (e.g. polychlorinated biphenyls [PCBs], polychlorinated dibenzodioxins and polychlorinated dibenzofurans [PCDDs/Fs]), and c) polyaromatic compounds (e.g. polycyclic aromatic hydrocarbons [PAHs]). The bioassay, which is based on EROD (7-ethoxyresorufin O-deethylase) induction in cultured chicken embryo livers detected dioxin-like activity in all unfractionated extracts and in the di- and polyaromatic fractions of all sludge extracts, but not in the monoatomatic/aliphatic fractions. The levels ranged between 6 and 109 pg bio-TEQ/g sludge (d.w.). In sediment samples from rural lakes in Sweden, levels of about 5 pg bio-TEQ/g (d.w.) have been found. The polyaromatic fractions of the sludge samples were potent in the bioassay, probably due to various PAHs and other polyaromatics in the sludge. The levels of six PAHs that are screened for in the sludge at Swedish STPs accounted for only 3-10% of the observed EROD-induction by the polyaromatic fractions. Consequently, many other polyaromatic EROD-inducing compounds were present in the sludge. Inclusion of a biological test like the chicken embryo liver bioassay in the screening of sludge would improve the ability to detect the presence of bioactive dioxin-like compounds. A theoretical estimation of bio-TEQ concentrations in farm-soil following long-term application of sludge with bio-TEQ concentrations similar to those observed in this investigation indicated that the bio-TEQ levels in soil would increase very slowly over time. The chicken embryo liver bioassay proved useful in assessing levels of dioxin-like compounds in sewage sludge and it gives valuable complementary information to chemical analysis data.

  • 13.
    Engwall, Magnus
    et al.
    Örebro University, School of Science and Technology. Department of Environmental Toxicology, Evolutionary Biology Center, Uppsala University, Uppsala, Sweden; .
    Hjelm, Katarina
    Department of Environmental Toxicology, Evolutionary Biology Center, Uppsala University, Uppsala, Sweden .
    Uptake of dioxin-like compounds from sewage sludge into various plant species: assessment of levels using a sensitive bioassay2000In: Chemosphere, ISSN 0045-6535, E-ISSN 1879-1298, Vol. 40, no 9-11, p. 1189-1195Article in journal (Refereed)
    Abstract [en]

    A bioassay for the detection of dioxin-like compounds was used to estimate uptake of dioxin-like compounds in carrots, oil seed rape seeds, zucchinis and cucumbers grown in soil amended with sewage sludge from Swedish sewage treatment plants (STP), This sensitive bioassay is based on 7-ethoxyresorufin O-deethylase (EROD)-induction in cultured chicken embryo livers and reflects the combined biological effect of all dioxin-like compounds in a sample, including ones that seldom are analyzed. The bioassay detected low concentrations of dioxin-like compounds in all carrot, zucchini and cucumber samples, but did not detect any dioxin-like compounds in the rape seeds. In carrots the concentrations were increased up to seven times when grown in soil amended with high applications of some of the sludge samples, while others did not increase the concentrations compared to control. More realistic sludge applications only increased the concentrations slightly. The sludge-fertilized carrots contained the highest concentrations of the investigated plants (up to 14 pg bioassay-derived TCDD equivalents (bio-TEQs)/g d.w.). In the carrots, differences in uptake of dioxin-like compounds depended on the sludge origin, which may be due to more easily bioaccumulated dioxin-like compounds in some sludge samples, or other components that facilitated uptake into the carrots. In the cucumbers, a more than two-fold increase (from 0.2 to 0.5 pg bio-TEQs/g d.w.) was observed in specimens grown in sludge-amended soil when compared to controls, suggesting a small uptake from the roots to the shoots. No sludge-dependent increase in uptake was seen in the zucchini fruits. The bio-TEQ levels were generally low in the consumable above ground plant parts of the investigated species. However, the question if repeated sludge application results in a soil accumulation of dioxin-like compounds, thereby increasing the risk of plant uptake, remains to be investigated.

  • 14.
    Engwall, Magnus
    et al.
    Örebro University, School of Science and Technology.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Bioanalys av organiska föroreningars biotillgänglighet: tillämpning i sanerade massor2009Report (Other (popular science, discussion, etc.))
    Abstract [sv]

    Polycykliska aromatiska kolväten (PAHer) är relativt vanliga i förorenadeområden, särskilt på gamla gasverktomter, bensinstationer och tidigare impregneringsanläggningar.På grund av deras toxicitet så är sanering av PAH-förorenadeområden av hög prioritet. För att minska riskerna med PAH-förorenade jordar,både före och efter sanering, är det viktigt att åstadkomma en heltäckande riskbedömningoch säker klassning av dessa jordmassor. De generella riktvärden förPAH-förorenad mark som används idag är i regel baserade på kemisk analys av 16standard PAHer (PAH16), trots att det ofta förekommer 100-tals PAHer och PAHmetaboliteri jordarna.I detta projekt har vi genom att jämföra kemisk och biologisk analys (H4IIEluc)av ett flertal sanerade PAH-förorenade jordprover studerat om toxicitetenverkligen minskar i proportion till minskningen av PAHer i jordarna. H4IIE-luc ären mekanismspecifik bioanalys som detekterar alla ämnen som aktiverar Ahreceptorn,en av de två viktigaste mekanismerna bakom PAHers toxicitet. Jämförelsenav resultaten visade att den totala toxiciteten i de sanerade jordprovernainte gick att förklara med kemisk analys av PAH16 och att man därmed med dagensanalysmetodik riskerar att missa toxikologiskt relevanta PAHer och andra liknandeämnen. Vidare kemiska identifieringsstudier samt bioanalytiska studier krävs föratt ta reda på om dessa okända ämnen utgör en risk för människa eller miljö.Våra resultat visar på svagheten med kemisk analys av ett mindre antal ämnensom grundval för klassning av renade massor. Det är därför är rimligt att inkluderamekanismspecifika tester i riskbedömning och vid klassning av renade PAHförorenadejordar. Dels för att minimera riskerna som dessa jordar kan utgöra förmänniskor och miljö, dels för att man med en större säkerhet och i större utsträckningskall kunna återanvända sanerade jordmassor.

  • 15.
    Engwall, Magnus
    et al.
    Department of Environmental Toxicology, Evolutionary Biology Center, Uppsala University, Uppsala, Sweden;.
    Näf, C.
    Aquatic Chemical Ecotoxiocology, Department of Zoology, Stockholm University, Stockholm, Sweden.
    Broman, D.
    Department of Zoology, Inst. of Appl. Environ. Research, Stockholm University, Sweden .
    Brunström, Björn
    Department of Environmental Toxicology, Evolutionary Biology Center, Uppsala University, Uppsala, Sweden;.
    Biological and chemical determination of contaminant levels in settling particulate matter and sediments: a Swedish river system before, during, and after dredging of PCB-contaminated lake sediments1998In: Ambio, ISSN 0044-7447, E-ISSN 1654-7209, Vol. 27, no 5, p. 403-410Article in journal (Refereed)
    Abstract [en]

    A sensitive bioassay, based on EROD induction in cultured chicken embryo livers, was used together with chemical analysis to determine levels of dioxin-like contaminants in particulate matter in Eman. Extracts of sediment and settling particulate matter (SPM) collected in the river system before, during, and after dredging of a PCB contaminated lake, Lake Jarnsjon, were separated into three fractions containing a) monoaromatic/aliphatic; b) diaromatic (e.g., PCBs and polychlorinated dibenzodioxins and dibenzofurans); and c) polyaromatic compounds (e.g., polycyclic aromatic hydrocarbons). The samples from Lake Jarnsjon contained the highest PCB concentrations and the diaromatic extracts from Lake Jarnsjon samples showed the highest activities of EROD-inducing diaromatic compounds. The dioxin-like activity of the diaromatic fraction in sediment collected after the dredging of Jarnsjon had only around 1% of the activity of the pre-dredging sediment, showing that the remediation was successful in terms of removal of the PCB-contaminated sediment from the lake. In SPM collected immediately downstream from Jarnsjon, levels of diaromatic dioxin-like compounds were elevated during the dredging, and decreased after that. The post-dredging concentrations were however higher than in SPM from lakes upstream of Jarnsjon, showing that elevated levels of dioxin-like diaromatic compounds were still present in the water System downstream of Jarnsjon a couple of years after the dredging.

  • 16.
    Engwall, Magnus
    et al.
    Örebro University, School of Science and Technology.
    Schnürer, A.
    Department of Microbiology, Swedish University of Agricultural Sciences, Uppsala, Sweden .
    Fate of Ah-receptor agonists in organic household waste during anaerobic degradation: estimation of levels using EROD induction in organ cultures of chick embryo livers2002In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 297, no 1-3, p. 105-108Article in journal (Refereed)
    Abstract [en]

    The fate of 7-ethoxyresorufin O-deethylase (EROD)-inducing compounds in source-separated organic household waste subject to anaerobic degradation (i.e. mesophilic/thermophilic anaerobic treatment) was investigated using organ cultures of embryonic chicken livers from fertilised hen eggs. This bioassay reflects the combined effect of all EROD-inducing, possibly dioxin-like compounds in a sample, including chemicals that are seldom or never analysed. All samples tested induced EROD in the bioassay, indicating the presence of dioxin-like compounds. In the anaerobic processes, the amounts of acid-resistant EROD-inducing compounds coming out of the reactors were considerably higher than the incoming amounts, especially for the low-temperature (mesophilic) process. This apparent production of EROD-inducing compounds may be due to de novo synthesis or to an increase in the EROD-inducing potency of the compounds in the material.

  • 17.
    Engwall, Magnus
    et al.
    Örebro University, School of Science and Technology.
    Venizelos, Nikolaos
    Örebro University, School of Health and Medical Sciences.
    Westman, Ola
    Örebro University, School of Science and Technology.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Nordén, Marcus
    Örebro University, School of Science and Technology.
    Hollert, Henner
    RWTH Aachen University.
    Johansson, Jessica
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Polycyclic aromatic hydrocarbons (PAHs) reduce hepatic beta-oxidation of fatty acids in chick embryos2013In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 20, no 3, p. 1881-1888Article in journal (Refereed)
    Abstract [en]

    Polycyclic aromatic hydrocarbons (PAHs) are widespread fused-ring contaminants formed during incomplete combustion of almost all kind of organic materials from both natural and anthropogenic sources. Some PAHs have been shown to be carcinogenic to humans, and a wide range of PAHs are found in wildlife all around the globe including avian species. The purpose of this project was to assess the effects of a standard mixture of 16 PAHs (United States Environmental Protection Agency) on the hepatic fatty acid beta-oxidation in chicken embryos (Gallus gallus domesticus) exposed in ovo. The hepatic beta-oxidation was measured using a tritium release assay with [9,10-H-3]-palmitic acid (16:0) as substrate. Treated groups were divided into groups of 0.05, 0.1, 0.3, 0.5, and 0.8 mg PAHs/kg egg weight. The hepatic beta-oxidation was reduced after exposure in ovo to the 16 PAHs mixture compared to control. The mechanisms causing reduced fatty acid oxidation in the present study are unclear, however may be due to deficient membrane structure, the functionality of enzymes controlling the rate of fatty acid entering into the mitochondria, or complex pathways connected to endocrine disruption. To the best of our knowledge, this is the first time a PAH-caused reduction of hepatic beta-oxidation of fatty acids in avian embryos has been observed. The implication of this finding on risk assessment of PAH exposure in avian wildlife remains to be determined.

  • 18.
    Gustavsson, L.
    et al.
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Genotoxic activity of nitroarene-contaminated industrial sludge following large-scale treatment in aerated and non-aerated sacs2006In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 367, no 2-3, p. 694-703Article in journal (Refereed)
    Abstract [en]

    An industrial sludge containing a complex mixture of nitroaromatic compounds was treated in industrial large-scale aerobic and anaerobic biodegradation processes, performed in compost sacs. The goal was to study changes in genotoxicity during the two different oxygen regimes using the umuC genotoxicity assay. The composting sac was actively aerated during 3 months and allowed to mature for another 3 months. The anaerobic sac was not aerated for 5 months and aerated during the last month in order to enhance degradation of remaining organic carbon. The sludge was obtained from the wastewater treatment plant at an industrial area in Karlskoga, Sweden. The biodegradation study was performed at a commercial waste treatment plant in Stockholm, according to the company routine procedure when treating household waste in sealed sacs.

    The material from the non-aerated system showed increased genotoxicity in the acetone-soluble fraction after treatment, as did the water-soluble fraction. The subsequent aeration period did not decrease the toxicity below the genotoxicity limit. The increase in the water-soluble genotoxic compounds may pose an environmental problem during secondary storage or use of sludge treated this way, since leakage of water-dissolved genotoxic compounds may occur.

    The composting process also generated genotoxicity, but this was restricted to acetone-soluble compounds, while the water-soluble compounds remained low in genotoxicity. The aerated process therefore seems more favorable in term of risk reduction of this industrial sludge, although it is necessary to optimize the aerated process in order to achieve non-toxic levels of potential genotoxic compounds extractable by organic solvents.

  • 19.
    Gustavsson, L.
    et al.
    Karlskoga Environment and Energy Company, Karlskoga, Sweden.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Treatment of sludge containing nitro-aromatic compounds in reed-bed mesocosms: Water, BOD, carbon and nutrient removal2012In: Waste Management, ISSN 0956-053X, E-ISSN 1879-2456, Vol. 32, no 1, p. 104-109Article in journal (Refereed)
    Abstract [en]

    Since the mid-1970s, Sweden has been depositing 1 million ton d.w sludge/year, produced at waste water treatment plants. Due to recent legislation this practice is no longer a viable method of waste management. It is necessary to improve existing and develop new sludge management techniques and one promising alternative is the dewatering and treatment of sludge in constructed wetlands. The aim of this study was to follow reduction of organic carbon, BOD and nutrients in an industrial sludge containing nitro-aromatic compounds passing through constructed small-scale wetlands, and to investigate any toxic effect such as growth inhibition of the common reed Phragmites australis. The result showed high reduction of all tested parameters in all the outgoing water samples, which shows that constructed wetlands are suitable for carbon and nutrient removal. The results also showed that P. australis is tolerant to xenobiotics and did not appear to be affected by the toxic compounds in the sludge. The sludge residual on the top of the beds contained low levels of organic carbon and is considered non-organic and could therefore be landfilled. Using this type of secondary treatment method, the amount of sludge could be reduced by 50-70%, mainly by dewatering and biodegradation of organic compounds.

  • 20.
    Gustavsson, Lillemor
    et al.
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Genotoxicity of nitroaromatic compounds in sludge after large scale biological treatment in aerated and nonaerated sacsManuscript (Other academic)
  • 21.
    Gustavsson, Lillemor
    et al.
    Örebro University, Department of Natural Sciences.
    Hollert, Henner
    Jönsson, Sofie
    Örebro University, Department of Natural Sciences.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Reed beds receiving industrial sludge containing nitroaromatic compounds: effects of outgoing water and bed material extracts in the umu-C genotoxicity assay, DR-CALUX assay and on early life stage development in Zebrafish (Danio rerio)Manuscript (Other academic)
  • 22.
    Gustavsson, Lillemor
    et al.
    Örebro University, Department of Natural Sciences.
    Hollert, Henner
    Jönsson, Sofie
    Örebro University, Department of Natural Sciences.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Reed beds receiving industrial sludge containing nitroaromatic compounds: Effects of outgoing water and bed material extracts in the umu-c genotoxicity assay, DR-CALUX assay and on early life stage development in Zebrafish (Danio rerio)2007In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 14, no 3, p. 202-211Article in journal (Refereed)
    Abstract [en]

    Goal, Scope and Background:  Sweden has prohibited the deposition of organic waste since January, 2005. Since 1 million tons of sludge is produced every year in Sweden and the capacity for incineration does not fill the demands, other methods of sludge management have to be introduced to a larger degree. One common method in the USA and parts of Europe is the use of wetlands to treat wastewater and sewage sludge. The capacity of reed beds to affect the toxicity of a complex mixture of nitroaromatics in sludge, however, is not fully elucidated. In this study, an industrial sludge containing explosives and pharmaceutical residues was therefore treated in artificial reed beds and the change in toxicity was studied. Nitroaromatic compounds, which are the main ingredients of many pharmaceuticals and explosives, are well known to cause cytotoxicity and genotoxicity. Recently performed studies have also showed that embryos of zebrafish (Danio rerio) are sensitive to nitroaromatic compounds. Therefore, we tested the sludge passing through constructed wetlands in order to detect any changes in levels of embryotoxicity, genotoxicity and dioxin-like activity (AhR-agonists). We also compared unplanted and planted systems in order to examine the impact of the root system on the fate of the toxicants. Methods:  An industrial sludge containing a complex mixture of nitroaromatics was added daily to small-scale constructed wetlands (vertical flow), both unplanted and planted with Phragmites australis. Sludge with an average dry weight of 1.25%, was added with an average hydraulic loading rate of 1.2 L/day. Outgoing water was collected daily and stored at −20°C. The artificial wetland sediment was Soxhlet extracted, followed by clean-up with multi-layer silica, or extracted by ultrasonic treatment, yielding one organic extract and one water extract of the same sample. Genotoxicity of the extracts was measured according to the ISO protocol for the umu-C genotoxicity assay (ISO/TC 147/SC 5/WG9 N8), using Salmonella typhimurium TA1535/pSK1002 as test organism. Embryotoxicity and teratogenicity were studied using the fish egg assay with zebrafish (Danio rerio) and the dioxin-like activity was measured using the DR-CALUX assay. Chemical analyses of nitroaromatic compounds were performed using Solid Phase Micro Extraction (SPME) and GC-MS. Results:  Organic extracts of the bed material showed toxic potential in all three toxicity tests after two years of sludge loading. There was a difference between the planted and the unplanted beds, where the toxicity of organic extracts overall was higher in the bed material from the planted beds. The higher toxicity of the planted beds could have been caused by the higher levels of total carbon in the planted beds, which binds organic toxicants, and by enrichment caused by lower volumes of outgoing water from the planted beds. Discussion:  Developmental disorders were observed in zebrafish exposed directly in contact to bed material from unplanted beds, but not in fish exposed to bed material from planted beds. Hatching rates were slightly lower in zebrafish exposed to outgoing water from unplanted beds than in embryos exposed to outgoing water from planted beds. Genotoxicity in the outgoing water was below detection limit for both planted and unplanted beds. Most of the added toxicants via the sludge were unaccounted for in the outgoing water, suggesting that the beds had toxicant removal potential, although the mechanisms behind this remain unknown. Conclusions:  During the experimental period, the beds received a sludge volume (dry weight) of around three times their own volume. In spite of this, the toxicity in the bed material was lower than in the sludge. Thus, the beds were probably able to actually decrease the toxicity of the added, sludge-associated toxicants. When testing the acetone extracts of the bed material, the planted bed showed a higher toxicity than the unplanted beds in all three toxicity tests. The toxicity of water extracts from the unplanted beds, detected by the fish egg assay, were higher than the water extracts from the planted beds. No genotoxicity was detected in outgoing water from either planted or unplanted beds. All this together indicates that the planted reed beds retained semi-lipophilic acetone-soluble toxic compounds from the sludge better than the unplanted beds, which tended to leak out more of the water soluble toxic compounds in the outgoing water. The compounds identified by SPME/GC in the outgoing water were not in sufficient concentrations to have caused induction in the genotoxicity test. Recommendations and Perspectives:  This study has pointed out the benefits of using constructed wetlands receiving an industrial sludge containing a complex mixture of nitroaromatics to reduce toxicity in the outgoing water. The water from planted, constructed wetlands could therefore be directed to a recipient without further cleaning. The bed material should be investigated over a longer period of time in order to evaluate potential accumulation and leakage prior to proper usage or storage. The plants should be investigated in order to examine uptake and possible release when the plant biomass is degraded.

  • 23.
    Gustavsson, Lillemor
    et al.
    Örebro University, Department of Natural Sciences.
    Klee, Nina
    Olsman, Helena
    Hollert, Henner
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Fate of Ah receptor agonists during biological treatment of an industrial sludge containing explosives and pharmaceutical residues2004In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 11, no 6, p. 379-386Article in journal (Refereed)
    Abstract [en]

    GOAL, SCOPE AND BACKGROUND:

    Sweden is meeting prohibition for deposition of organic waste from 2005. Since 1 million tons of sludge is produced every year in Sweden and the capacity for incineration does not fill the demands, other methods of sludge management have to be introduced to a higher degree. Two biological treatment alternatives are anaerobic digestion and composting. Different oxygen concentrations result in different microbial degradation pathways and, consequently, in a different quality of the digestion or composting residue, It is therefore necessary to study sludge treatment during different oxygen regimes in order to follow both degradation of compounds and change in toxicity. In this study, an industrial sludge containing explosives and pharmaceutical residues was treated with anaerobic digestion or composting, and the change in toxicity was studied. Nitroaromatic compounds, which are the main ingredients of both pharmaceutical and explosives, are well known to cause cytotoxicity and genotoxicity. However, little data are available concerning sludge with nitroaromatics and any associated dioxin-like activity. Therefore, we studied the sludge before and after the treatments in order to detect any changes in levels of Ah receptor (AhR) agonists using two bioassays for dioxin-like compounds.

    METHODS:

    An industrial sludge was treated with anaerobic digestion or composting in small reactors in a semi-continuous manner. The same volume as the feeding volume was taken out daily and stored at -20 degrees C. Sample preparation for the bioassays was done by extraction using organic solvents, followed by clean up with silica gel or sulphuric acid, yielding two fractions. The fractions were dissolved in DMSO and tested in the bioassays. The dioxin-like activity was measured using the DR-CALUX assay with transfected H4IIE rat hepatoma pGudluc cells and an EROD induction assay with RTL-W1 rainbow trout liver cells.

    RESULTS AND DISCUSSION:

    The bioassays showed that the sludge contained AhR agonists at levels of TCDD equivalents (TEQs) higher than other sludge types in Sweden. In addition, the TEQ values for the acid resistant fractions increased considerably after anaerobic digestion, resulting in an apparent formation of acid resistant TEQs in the anaerobic reactors. Similar results have been reported from studies of fermented household waste. There was a large difference in effects between the two bioassays, with higher TEQ levels in the RTL-W1 EROD assay than in the DR-CALUX assay. This is possibly due to a more rapid metabolism in rat hepatocytes than in trout hepatocytes or to differences in sensitivities for the AhR agonists in the sludge. It was also demonstrated by GC/FID analysis that the sludge contained high concentrations of nitroaromatics. It is suggested that nitroaromatic metabolites, such as aromatic amines and nitroanilines, are possible candidates for the observed bioassay effects. It was also found that the AhR agonists in the sludge samples were volatile.

    CONCLUSIONS:

    The sludge contained fairly high concentrations of volatile AhR agonists. The increase of acid resistant AhR agonist after anaerobic digestion warrants further investigations of the chemical and toxic properties of these compounds and of the mechanisms behind this observation.

    RECOMMENDATION AND OUTLOOK:

    This study has pointed out the benefits of using different types of mechanism-specific bioassays when evaluating the change in toxicity by sludge treatment, in which measurement of dioxin-like activity can be a valuable tool. In order to study the recalcitrant properties of the compounds in the sludge using the DR-CALUX assay, the exposure time can be varied between 6 and 24 hours. The properties of the acid-resistant AhR agonists formed in the anaerobic treatment have to be investigated in order to choose the most appropriate method for sludge management.

  • 24.
    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.

  • 25.
    Hagberg, Jessika
    et al.
    Örebro University, Department of Natural Sciences.
    Olsman, Helena
    Örebro University, Department of Natural Sciences.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Chemical and toxicological characterisation of PBDFs from photolytic decomposition of decaBDE in tolueneManuscript (Other academic)
  • 26.
    Hagberg, Jessika
    et al.
    Örebro University, Department of Natural Sciences.
    Olsman, Helena
    Örebro University, Department of Natural Sciences.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Lindström, Gunilla
    Örebro University, Department of Natural Sciences.
    Chemical and toxicological characterisation of PBDFs from photolytic decomposition of decaBDE in toluene2006In: Environment International, ISSN 0160-4120, E-ISSN 1873-6750, Vol. 32, no 7, p. 851-857Article in journal (Refereed)
    Abstract [en]

    A substantial formation of PBDF congeners was observed during photolytic decomposition of decaBDE in toluene. The decaBDE degradation was monitored by chemical and toxicological analysis and in all, twenty-seven mono- to hexasubstituted polybrominated dibenzofurans (PBDFs) were detected in toluene solutions of decabromodiphenyl ether (decaBDE) after irradiation with UV-A, UV-AB and UV-ABC. The concentration levels of PBDFs formed after 16 h of UV exposure increased with wider spectra and were determined to be 3.5, 4.2 and 14 microg/ml after UV-A, UV-AB and UV-ABC irradiation, respectively. In accordance, bioassay derived TEQs (bio-TEQs), determined with the DR-CALUX assay, increased with a similar pattern. The PBDFs formed after the three UV exposures accounted for 0.31%, 0.35% and 1.2% of the initial amount of decaBDE (molar basis). The PBDF congener patterns were consistent in all three UV experiments which imply that no alterations were induced in the PBDF formation or degradation processes due to differences in UV irradiation. However, these processes tended to increase with wider spectra and increasing radiation energy most likely due to the strong absorbance of for example decaBDE at shorter wavelengths. After total decaBDE decomposition the PBDF formation increased significantly in the UV-ABC experiment. The tetra to hexasubstituted BDFs constituted the majority of detected compounds in all experiments. In all samples, the estimated chemical TEQ indicate that the bio-TEQs observed are largely explained by the presence of non-2,3,7,8-substituted PBDFs with relatively low toxicological potencies.

  • 27.
    Henriksson, Sara
    et al.
    Örebro University, School of Science and Technology. 15 Environmental Staff, Arvika, Sweden.
    Bjurlid, Filip
    Örebro University, School of Science and Technology.
    Rotander, Anna
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Lindström, Gunilla
    Örebro University, School of Science and Technology.
    Westberg, Håkan
    Örebro University, School of Science and Technology.
    Hagberg, Jessika
    Örebro University, School of Science and Technology.
    Uptake and bioaccumulation of PCDD/Fs in earthworms after in situ and in vitro exposure to soil from a contaminated sawmill site2017In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 580, p. 564-571Article in journal (Refereed)
    Abstract [en]

    Uptake of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) was studied in earthworms collected from a sawmill site in Sweden with severe PCDD/Fs contamination (the hot spot concentration was 690,000 ng TEQWHO2005/kg d.w.) in order to investigate the transfer of PCDD/Fs from the site to the biota. PCDD/Fs concentrations in the collected earthworms were compared to PCDD/Fs concentrations in laboratory exposed earthworms (Eisenia fetida), which were exposed to contaminated soils from the sawmill site for 34 days. All analyses were performed by high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS). PCDD/Fs concentrations in the earthworms ranged from 290 to 520,000 pg/g (f.w.). The main congeners found in both soils and earthworms were OCDF, 1234678-HpCDF, OCDD and 1234678-HpCDD. The study showed that the PCDD/Fs in the soil were biovailable to the earthworms and the PCDD/Fs concentrations in the soils correlated with the concentrations in the earthworms. Earthworm samples from soil with lower concentration had higher bioaccumulation factors than samples from soils with high concentration of contamination. Thus, a less contaminated soil could yield higher concentrations in earthworms compared to a higher contaminated soil. Assuming that when assessing risks with PCDD/F contaminated soil, a combination of chemical analysis of soil PCDD/Fs concentrations and bioavailability should be employed for a more comprehensive risk assessment.

  • 28. Hinger, Gunnar
    et al.
    Brinkmann, Markus
    Bluhm, Kerstin
    Sagner, Anne
    Takner [Olsman], Helena
    Örebro University, School of Science and Technology.
    Eisentraeger, Adolf
    Braunbeck, Thomas
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Tiehm, Andreas
    Hollert, Henner
    Some heterocyclic aromatic compounds are Ah receptor agonists in the DR-CALUX assay and the EROD assay with RTL-W1 cells2011In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 18, no 8, p. 1297-1304Article in journal (Refereed)
    Abstract [en]

    Purpose Heterocyclic aromatic compounds containing nitrogen, sulfur, or oxygen heteroatoms (NSO-HET) have been detected in air, soil, marine, and freshwater systems. However, only few publications are available investigating NSO-HET using in vitro bioassays. To support better characterization of environmental samples, selected NSO-HET were screened for dioxin-like activity in two bioassays. Methods The present study focuses on the identification and quantification of dioxin-like effects of 12 NSO-HET using the DR-CALUX assay, and the 7-ethoxyresorufin-O-deethylase (EROD) assay with the permanent fish liver cell line RTL-W1. Changes of the total medium compound concentrations during the test procedure due to, e.g., sorption or volatilization were quantified using GC/MS. Results The NSO-HET benzofuran, 2,3-dimethylbenzofuran, dibenzofuran, dibenzothiophen, acridine, xanthene, and carbazole caused a response in the DR-CALUX assay. Only benzofuran and 2,3-dimethylbenzofuran were also positive in the EROD assay. All other compounds were inactive in the EROD assay. Relative potency (REP) values ranged from (2.80 +/- 1.32) center dot 10(-8) to (3.26 A +/- 2.03) A center dot 10(-6) in the DR-CALUX and from (3.26 A +/- 0.91) A center dot 10(-7) to (4.87 A +/- 1.97) A center dot 10(-7) in the EROD assay. Conclusions The REP values were comparable to those of larger polycyclic aromatic hydrocarbons, e.g., fluoranthene and pyrene. Thus, and because of the ubiquitous distribution of heterocyclic aromatic compounds in the environment, the provided data will further facilitate the bioanalytical and analytical characterization of environmental samples towards these toxicants.

  • 29.
    Hollert, Henner
    et al.
    Department of Zoology, University of Heidelberg, Im Neuenheimer Feld 230, Heidelberg, Germany .
    Dürr, M.
    Department of Hygiene, Halle, Germany .
    Olsman, Helena
    Örebro University, School of Science and Technology.
    Halldin, Krister
    Department of Environmental Toxicology, Uppsala, Sweden .
    van Bavel, Bert
    Örebro University, School of Science and Technology. Department of Chemistry, Environmental Chemistry, Umeå, Sweden .
    Brack, W.
    UFZ Centre for Environmental Research Leipzig-Halle, Leipzig, Germany .
    Tysklind, M.
    Department of Environmental Toxicology, Uppsala, Sweden .
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Braunbeck, T.
    Department of Hygiene, Halle, Germany .
    Biological and chemical determination of dioxin-like compounds in sediments by means of a sediment triad approach in the catchment area of the River Neckar2002In: Ecotoxicology, ISSN 0963-9292, E-ISSN 1573-3017, Vol. 11, no 5, p. 323-336Article in journal (Refereed)
    Abstract [en]

    To evaluate the sediment quality of selected sites in the catchment area of the River Neckar, an integrative assessment approach was used to assess the ecological hazard potential of dioxin-like sediment compounds. The approach is based on 7-ethoxyresorufin-O-deethylase (EROD) induction in embryonic chicken liver culture and comprehensive chemical analyses of polycyclic aromatic hydrocarbons (priority PAHs according to the US Environmental Protection Agency). The majority of the sediment extracts exhibited high potencies as EROD-inducers. In one sediment sample, which was influenced by a sewage treatment plant, a very high concentration of 930 ng bioassay 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalents (bio-TEQs)/g organic carbon could be determined. However, in none of the samples, more than 6% of the EROD-inducing potency could be explained by the PAHs analyzed chemically. Thus, non-analyzed compounds with EROD-inducing potency were present in the extracts. A fractionation of sediment samples according to pH allowed to localize the major part of EROD-inducing compounds in the neutral fractions. However, a significant portion of the EROD induction could also be explained by the acidic fractions. Following the concept of the Sediment Quality Triad according to Chapman, in situ alterations of macrozoobenthos were examined. A comparison of the results predicted by the EROD assay and chemical analyses with alterations in situ, as measured by means of the saprobic index and the ecotoxicological index according to Carmargo, revealed a high ecological relevance of the results of bioassays and chemical analyses for major sites.

  • 30. 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.

  • 31.
    Håkansson, H.
    et al.
    Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
    Sundin, P.
    Chemical Ecology and Ecotoxicology, Department of Ecology, Lund University, Lund, Sweden.
    Andersson, T.
    Department of Zoophysiology, University of Göteborg, Göteborg, Sweden.
    Brunström, B.
    Department of Zoophysiology, Uppsala University, Uppsala, Sweden.
    Dencker, L.
    Department of Toxicology, Uppsala University, Uppsala, Sweden.
    Engwall, Magnus
    Department of Zoophysiology, Uppsala University, Uppsala, Sweden.
    Ewald, G.
    Chemical Ecology and Ecotoxicology, Department of Ecology, Lund University, Lund, Sweden.
    Gilek, M.
    Department of Zoology, Stockholm University, Stockholm, Sweden.
    Holm, G.
    Department of Zoology, Stockholm University, Stockholm, Sweden; Swedish Environmental Research Institute, Stockholm, Sweden.
    Honkasalo, S.
    Institute of Environmental Medicine, Karolinska Institutet, S-10401 Stockholm, Sweden.
    Idestam-Almquist, J.
    Department of Botany, Stockholm University, Stockholm, Sweden.
    Jonsson, P.
    Swedish Environmental Protection Agency, Solna, Sweden.
    Kautsky, N.
    Department of Systems Ecology, Stockholm University, Stockholm, Sweden.
    Lundberg, G.
    Swedish Environmental Research Institute, Stockholm, Sweden.
    Lund-Kvernheim, A.
    Center for Industrial Research, Oslo, Norway.
    Martinsen, K.
    Center for Industrial Research, Oslo, Norway.
    Norrgren, L.
    Swedish Environmental Research Institute, Stockholm, Sweden; Department of Pathology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Personen, M.
    Department of Zoophysiology, University of Göteborg, Göteborg, Sweden.
    Rundgren, M.
    Department of Toxicology, Karolinska Institutet, Stockholm, Sweden.
    Stålberg, M.
    Institute of Environmental Medicine, Karolinska Institutet, S-10401 Stockholm, Sweden.
    Tarkpea, M.
    Swedish Environmental Protection Agency, Aquatic Toxicology Section, Nyköping, Sweden.
    Wesén, C.
    Department of Technical Analytical Chemistry, Chemical Center, Lund University, Lund, Sweden.
    In vivo and in vitro toxicity of fractionated fish lipids, with particular regard to their content of chlorinated organic compounds1991In: Pharmacology and Toxicology, ISSN 0901-9928, E-ISSN 1600-0773, Vol. 69, no 6, p. 459-471Article in journal (Refereed)
    Abstract [en]

    Six different lipid matrices (the intact lipid (IL), four lipid fractions with different polarity, and the free fatty acids (FFAs) obtained by hydrolysis of the triacylglycerol (TAG) containing fraction) were obtained from salmon (Salmo salar) and eel (Anguilla anguilla), each collected at a contaminated and a comparatively uncontaminated catch site along the coast of Scandinavia. The lipid matrices were studied in toxicological test systems representing various biological functions of different organ systems from several species and trophic levels. The results were evaluated with particular respect to the concentrations of extractable organically bound chlorine (EOCl) in the matrices tested. In some test systems, the specimens with a higher EOCl concentration appeared to be more toxic. For example, the TAG containing fraction (F2) from Idefjord eel, having a higher EOCl content than F2 from Oslofjord eel, reduced the number and hatchability of eggs laid by zebrafish. Both IL and F2 of Idefjord eel increased mortality and reduced the oxygen/nitrogen-ratio in blue mussels. Non-polar compounds (F1) from Bothnian Sea salmon induced 7-ethoxyresurofin O-deethylase (EROD) activity in rainbow trout hepatocytes, whereas F1 from Senja salmon did not. F1 from Bothnian Sea salmon also reduced the number of T-cells in foetal mouse thymus anlagen in vitro compared with the cell number in anlagen exposed to F1 from Senja salmon. A positive correlation between EOCl concentration and test response was found for EROD activity in rainbow trout hepatocytes and for ATP-leakage in Erlich ascites tumour cells when testing the phospolipid containing fraction (F4). However, in most test systems the fish oils, irrespective of EOCl content, were of low toxicity, and the observed effects need to be verified in future studies.

  • 32.
    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.

  • 33.
    Julander, Anneli
    et al.
    Örebro University, Department of Natural Sciences.
    Karlsson, Marie
    Örebro University, Department of Natural Sciences.
    Hagström, K.
    Örebro University, Department of Natural Sciences.
    Ohlson, C.-G.
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Bryngelsson, I.-L.
    Westberg, Håkan
    Örebro University, Department of Natural Sciences.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Polybrominated diphenyl ethers: plasma levels and thyroid status of workers at an electronic recycling facility2005In: International Archives of Occupational and Environmental Health, ISSN 0340-0131, E-ISSN 1432-1246, Vol. 78, no 7, p. 584-592Article in journal (Refereed)
    Abstract [en]

    Objectives: Personnel working with electronic dismantling are exposed to polybrominated diphenyl ethers (PBDEs), which in animal studies have been shown to alter thyroid homeostasis. The aim of this longitudinal study was to measure plasma level of PBDEs in workers at an electronic recycling facility and to relate these to the workers’ thyroid status. Methods: PBDEs and three thyroid hormones: triiodothyronine (T3), thyroxin (T4) and thyroid stimulating hormone (TSH) were repeatedly analysed in plasma from 11 workers during a period of 1.5 years.Results: Plasma levels of PBDEs at start of employment were <0.5–9.1 pmol/g lipid weight (l.w.). The most common congener was PBDE #47 (median 2.8 pmol/g l.w.), followed by PBDE #153 (median 1.7 pmol/g l.w.), and PBDE #183 had a median value of <0.19 pmol/g l.w. After dismantling the corresponding median concentrations were: 3.7, 1.7 and 1.2 pmol/g l.w., respectively. These differences in PBDE levels were not statistically significant. PBDE #28 showed a statistically significantly higher concentration after dismantling than at start of employment (P=0.016), although at low concentrations (start 0.11 pmol/g l.w. and dismantling 0.26 pmol/g l.w.). All measured levels of thyroid hormones (T3, T4 and TSH) were within the normal physiological range. Statistically significant positive correlations were found between T3 and #183 in a worker, between T4 and both #28 and #100 in another worker and also between TSH and #99 and #154 in two workers. Conclusions: The workers’ plasma levels of PBDEs fluctuated during the study period. Due to small changes in thyroid hormone levels it was concluded that no relevant changes were present in relation to PBDE exposure within the workers participating in this study.

  • 34.
    Julander, Anneli
    et al.
    Örebro University, Department of Technology.
    Karlsson, Marie
    Örebro University, Department of Natural Sciences.
    Hagström, Katja
    Ohlson, Carl Göran
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Bryngelsson, Ing-Liss
    Westberg, Håkan
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Polybrominated diphenylethers and thyroid hormone status in human plasma of workers at electronic recycling facilityManuscript (preprint) (Other academic)
  • 35.
    Julander, Anneli
    et al.
    Örebro University, Department of Natural Sciences.
    Westberg, Håkan
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    van Bavel, Bert
    Örebro University, Department of Natural Sciences.
    Distribution of brominated flame retardants in different dust fractions in air from an electronics recycling facilityManuscript (preprint) (Other academic)
  • 36.
    Julander, Anneli
    et al.
    Örebro University, School of Science and Technology.
    Westberg, Håkan
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    van Bavel, Bert
    Örebro University, School of Science and Technology.
    Distribution of brominated flame retardants in different dust fractions in air from an electronics recycling facility2005In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 350, no 1-3, p. 151-160Article in journal (Refereed)
    Abstract [en]

    Twelve air samples were collected from an electronic recycling facility in Sweden representing three different dust fractions; respirable, total and inhalable dust. Four samples were collected from each fraction. The highest concentration of polybrominated diphenyl ether (PBDE) #209 (ten bromine atoms) was found in the samples from the inhalable dust fraction (ID), which was 10 times higher than for the "total dust" fraction (TD). The concentration ranges were 157.6-208.6; 13.9-16.7; and 2.8-3.3 ng/m3 for inhalable, total and respirable fractions, respectively. The second most abundant PBDE congener was PBDE #183 (seven bromine atoms), followed by the second most abundant substance 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) in all samples. In addition, decabromodiphenyl ethane (DeBDethane) was tentatively identified in five of the samples. Because of the large differences in air concentrations between the three fractions in ID, TD and RD, it is suggested that the inhalable instead of "total dust" fraction should be used to assess air concentrations, in particular for the larger and higher brominated flame retardants (BFRs).

  • 37.
    Kalbin, Georgi
    et al.
    Örebro University, Department of Natural Sciences.
    Li, Shaoshan
    Örebro University, Department of Natural Sciences.
    Olsman, Helena
    Örebro University, Department of Natural Sciences.
    Pettersson, Mikael
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Strid, Åke
    Örebro University, Department of Natural Sciences.
    Effects of UV-B in biological and chemical systems: equipment for wavelength dependence determination2005In: Journal of Biochemical and Biophysical Methods, ISSN 0165-022X, E-ISSN 1872-857X, Vol. 65, no 1, p. 1-12Article in journal (Refereed)
    Abstract [en]

    The thinning of the stratospheric ozone layer has prompted a large number of studies of UV-B-induced effects in biological and chemical systems. The wavelength dependency of such effects is of interest from mechanistic, physiological or economic points of view. Here, we describe an apparatus for determining the wavelength dependency of UV-B effects in biological and chemical systems. The apparatus consists of a high intensity UV radiation source and narrow bandpass filters to produce UV radiation in even intervals (between 280 and 360 nm). The usefulness of the equipment is demonstrated in two different systems: 1) Chalcone synthase (CHS) gene is up-regulated by UV-B radiation. Therefore quantitative analysis of the CHS gene expression was chosen in the present investigation for studies of the wavelength dependency of gene expression regulation in plants. Maximum induction of CHS expression was found at 300 nm with a 12-fold induction compared with the control; 2) The wavelength dependency of formation of dioxin-like photoproducts from the brominated flame retardant decabrominated diphenyl ether (DeBDE) is described. This is an example of UV-B-induced conversion of non-toxic species into a number of products of which some may be toxic in the environment. In the UV interval studied, the highest dioxin-like activity was found in the sample irradiated at 330 nm and therefore this wavelength is most important for the mechanism involved in photoconversion of DeBDE.

  • 38. 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.

  • 39.
    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.

  • 40.
    Keiter, Su
    et al.
    University of Heidelberg, Heidelberg, Germany.
    Baumann, L.
    University of Heidelberg, Heidelberg, Germany.
    Faerber, H.
    University of Bonn, Bonn, Germany.
    Holbech, H.
    University of Southern Denmark, Odense M, Denmark.
    Skutlarek, D.
    University of Bonn, Bonn, Germany.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Braunbeck, T.
    University of Heidelberg, Heidelberg, Germany.
    Long-term effects of a binary mixture of perfluorooctane sulfonate (PFOS) and bisphenol A (BPA) in zebrafish (Danio rerio)2012In: Aquatic Toxicology, ISSN 0166-445X, E-ISSN 1879-1514, Vol. 118-119, p. 116-129Article in journal (Refereed)
    Abstract [en]

    Previous in vitro studies have reported the potential of perfluorooctane sulfonate (PFOS) to increase the toxicity of other compounds. Given the complex nature of mixtures of environmental pollutants in aquatic systems together with the persistent and bioaccumulative properties of PFOS, this study aimed at evaluating the long-term effects and toxicity-increasing behavior of PFOS in vivo using the zebrafish (Danio rerio). Fish were maintained in flow-through conditions and exposed to single and binary mixtures of PFOS and the endocrine disruptor bisphenol A (BPA) at nominal concentrations of 0.6, 100 and 300 mu g/L and 10, 200 and 400 mu g/L, respectively. F1 and F2 generations were evaluated from 0 to 180 days post-fertilization (dpf) and F3 generation was evaluated from 0 to 14 dpf. Survival was documented in all generations, whereas growth, fecundity, fertilization rate, histological alterations (in liver, thyroid and gonads) and vitellogenin (Vtg) induction in males were evaluated for Fl and F2 generations. Data for growth were collected at 30, 90 and 180 dpf and data for histological evaluations and Vtg induction were analyzed at 90 and 180 dpf. No significant effects on survival were seen in the Fl generation in any treatment following 180 d exposure: however, in the F2 generation, 300 mu g/L PFOS both alone and in combination with BPA (10, 200 and 400 mu g/L) induced 100% mortality within 14 dpf. PFOS (0.6 and 300 mu g/L) did not increase the Vtg-inducing potential of BPA (10, 200 and 400 mu g/L) in a binary mixture. In contrast, binary mixtures with 300 mu g/L PFOS suppressed the Vtg levels in Fl males at 90 dpf when compared to single BPA exposures. Whereas the lowest tested PFOS concentration (0.6 mu g/L) showed an estrogenic potential in terms of significant Vtg induction, Vtg levels were generally found to decrease with increasing PFOS-exposure in both Fl and F2 generations. In Fl generation, BPA-exposure was found to increase Vtg levels in a concentration-dependent manner. Histological analyses of Fl and F2 fish revealed hepatocellular vacuolization, predominantly in males, following PFOS-exposure both alone and in combination with BPA. Hepatotoxicity by PFOS might explain the suppressed Vtg response seen in PFOS-exposed Fl and F2 males. PFOS-exposed fish also showed granulomas, mainly in the liver. Given previous reports of the immunosuppressive potential of PFOS, the granulomas could be a consequence of a PFOS-induced reduction of the immune response potential. In conclusion, the hypothesis that the presence of PFOS increases the endocrine potential of BPA could not be confirmed in zebrafish. Adverse effects on liver structure and survival were only seen at concentrations well above ecologically relevant concentrations; however, the decline in survival rates following PFOS-exposure seen over generations again documents the importance of long-term studies for the investigation of persistent environmental pollutants.

  • 41.
    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.

  • 42. Klee, Nina
    et al.
    Gustavsson, Lillemor
    Örebro University, Department of Natural Sciences.
    Kosmehl, Thomas
    Engwall, Magnus
    Örebro University, Department of Natural Sciences.
    Erdinger, Lothar
    Braunbeck, Thomas
    Hollert, Henner
    Changes in toxicity and genotoxicity of industrial sewage sludge samples containing nitro- and amino-aromatic compounds following treatment in bioreactors with different oxygen regimes2004In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 11, no 5, p. 313-320Article in journal (Refereed)
    Abstract [en]

    GOALS, SCOPE AND BACKGROUND: From 2005, deposition of organic waste will be banned in Sweden. Likewise, in Germany and Austria, similar bans are being planned, and further countries will probably follow. Thus, there is a need to develop new methods and to refine established techniques for sludge management in the whole of the European Union. For this end, there is also an urgent need for appropriate ecotoxicological approaches to elucidate and assess the hazard potential of sewage sludge. Therefore, the present study was designed to assess the capacity of various established sludge treatment methods using different oxygen regimes to degrade recalcitrant nitro-substituted organic compounds and reduce their toxicity. Sewage sludge samples from a wastewater treatment plant in Sweden (Cambrex Karlskoga AB, industrial area Björkborn) receiving wastewater from industries manufacturing pharmaceutical substances, chemical intermediates and explosives were processed with different sludge treatment methods. Among other treatment methods, bioreactors (for anaerobic and aerobic sludge treatment) were used. In the present investigation, a battery of in vitro bioassays was employed to compare the cytotoxic and genotoxic potentials of different fractions of sludge samples in order to elucidate whether the treatments were suitable to reduce the toxicity of the sludge.

    METHODS:

    In order to investigate the cytotoxicity of the extracts of treated and untreated sludge samples, the acute cytotoxicity test with the permanent cell line RTL-W1 was used. Genotoxicity was tested by means of the comet assay (single cell gel electrophoresis) with RTL-W1 cells, and mutagenicity was assessed with the Ames test using the Salmonella typhimurium strains TA98, TA98NR and TA100. Sludge toxicity was tested in different fractions of organic extracts produced by acetone and hexane extractions. The subsequent clean-up procedure (silica gel chromatography and elution with hexane and dichloromethane) resulted in two fractions, a lipophilic hexane-fraction and a semi-lipophilic dichloromethane-fraction. For the genotoxicity and mutagenicity tests, these fractions were reunited at equal ratios.

    RESULTS AND DISCUSSION:

    The acute cytotoxicity test with RTL-W1 cells revealed a high cytotoxic potential for the semi-lipophilic DM-fractions of all sludge samples with NR50 values (= effective concentration for 50% cell death in the neutral red test) from 8.9 up to 20 mg sludge d.w./ml medium. A low cytotoxic potential for the hexane fractions of the untreated sludge samples (NR50 400 to > 400 mg sludge d.w./ml medium) was observed, whereas the hexane fractions of the treated sludge samples showed elevated cytotoxicity increasing further with treatment in the bioreactors. The comet assay indicated that three out of eight of the reunited fractions had a significant genotoxic potential. Whereas the genotoxic potential of one sample treated anaerobically was very high with an induction factor of 11.6, a similar sample (taken from the same anaerobic reactor four months later) and one untreated sample showed lower potentials. The samples treated in another anaerobic bioreactor as well as the samples treated aerobically showed no genotoxic potential. Results indicate that aerobic treatment was basically adequate for reducing the genotoxicity of the sludge, whereas anaerobic treatment was only partly useful for reduction of genotoxicity. The Ames test revealed a very high mutagenic potential for the reunited fractions of the untreated sludge samples with strain TA98 (maximum induction factors (IFmax) up to 45) and a relatively high potential for one of the samples treated aerobically (S2, IFmax = 18 (TA98, S9-)), thus documenting the suitability of both anaerobic and aerobic treatments to reduce the mutagenicity of the samples, however, with the aerobic treatment being less effective.

    CONCLUSIONS:

    Overall, none of the microbiological treatments for wastewater sludge in bioreactors was found to be ideal for general toxicity reduction of the sludge samples. Whereas cytotoxicity of the sludge increased or levelled off in most cases following either treatment, genotoxicity both increased or decreased after anaerobic treatment, depending on the specific sample. However, mutagenicity could generally be reduced by anaerobic treatment and, to a lesser degree, by aerobic treatment.

    RECOMMENDATIONS AND PERSPECTIVES:

    The complex modification of the diverse damage potentials of sludge sample extracts by use of an in vitro biotest battery following treatment for toxicity reduction in bioreactors showed that considerations of different toxicological endpoints is essential for an adequate hazard assessment. Whereas in the case of cytotoxicity reduction, the reactors proved ineffective, mutagenicity could be reduced significantly at least in some cases in this case study.

  • 43.
    Kärrman, Anna
    et al.
    Örebro University, School of Science and Technology.
    Schönlau, Christine
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Exposure and effects of microplastics on wildlife: A review of existing data2016Report (Other academic)
  • 44.
    Lam, Monika M.
    et al.
    Örebro University, School of Science and Technology.
    Bülow, Rebecca
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Giesy, John P.
    University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Methylated PACs are more potent than their parent compounds: a study on AhR-mediated activity, degradability and mixture interactions in the H4IIE-luc assayManuscript (preprint) (Other academic)
  • 45.
    Lam, Monika M.
    et al.
    Örebro University, School of Science and Technology.
    Bülow, Rebecca
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Giesy, John P.
    Department of Veterinary Biomedical Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Methylated PACs Are More Potent than Their Parent Compounds-A Study on AhR-mediated Activity, Degradability and Mixture Interactions in the H4IIE-luc Assay2018In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618Article in journal (Refereed)
    Abstract [en]

    In this study, 26 polycyclic aromatic compounds (PACs) including native polycyclic aromatic hydrocarbons (PAHs), hydroxylated PAHs, alkylated and oxygenated PAHs and (alkylated) heterocyclic compounds were investigated for their aryl hydrocarbon receptor (AhR)-mediated potencies in the H4IIE-luc bioassay. Potential degradabilities of PACs were investigated by use of various durations of exposure (24, 48 or 72 h). Furthermore, various mixtures of PACs including PAHs, alkylated and oxygenated PAHs and heterocyclic compounds were tested for their joint AhR-mediated potency. Additive behaviors of PACs in mixtures was studied by comparing observed mixture potencies to mixture potencies predicted by use of the concentration addition (CA) model. Methylated derivatives were more potent than their parent compounds in the H4IIE-luc assay. A time-dependent decrease in relative potency was observed for all AhR-active compounds, which may be indicative of in vitro biotransformation. Monomethylated compounds seemed to be more rapidly transformed than analogous unsubstituted compounds. In addition, results of this study showed that the predictive power of the CA model increased with the number of compounds, suggesting additivity in multicomponent mixtures. Due to greater potency of methylated derivatives and their ubiquitous occurrence, there is a need for further research on the toxicity and mixture behavior of those environmentally and toxicologically relevant compounds.

  • 46.
    Lam, Monika M.
    et al.
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Denison, Michael S.
    University of California, Davis, CA, USA.
    Giesy, John P.
    University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Polyoxymethylene (POM) is a suitable tool for effect-based hazard assessment of PAC-contaminated soilManuscript (preprint) (Other academic)
  • 47.
    Lam, Monika M.
    et al.
    Örebro University, School of Science and Technology.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Denison, Michael S
    Department of Environmental Toxicology, University of California, Davis CA, USA.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Methylated polycyclic aromatic hydrocarbons and/or their metabolites are important contributors to the overall estrogenic activity of polycyclic aromatic hydrocarbon-contaminated soils2018In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 37, no 2, p. 385-397Article in journal (Refereed)
    Abstract [en]

    In the present study 42 polycyclic aromatic compounds (PACs) were investigated for their estrogenic potential using the VM7Luc4E2 transactivation assay. Relative potencies were determined for mass-balance analysis. In addition, compounds were tested in combination with the estrogen receptor (ER) antagonist vertical bar C vertical bar 182,780 (vertical bar C vertical bar) and the aryl hydrocarbon receptor antagonist/CYP1A1 inhibitor a-naphthoflavone. Luciferase induction and CYP1A1-dependent ethoxyresorufin-O-deethylase (EROD) activity were measured to assess whether the estrogenic activity was elicited by the compound itself and/or by its metabolites. Relative potencies ranged between 10(-7) and 10(-4). The ability of ICI to decrease luciferase activity stimulated by all compounds indicated that the induction responses were ER-dependent. The aryl hydrocarbon receptor antagonist/CYP1A1 inhibitor a-naphthoflavone decreased luciferase induction and EROD activity by several compounds, including the methylated chrysenes, suggesting that metabolites of these chemicals contributed to ER activation. Several PACs, such as acridine and its derivatives, appear to directly activate the ER. Furthermore, extracts of soils from industrial areas were examined using this bioassay, and estrogenic activity was detected in all soil samples. Mass-balance analysis using a combination of relative potencies and chemical analysis of the samples suggested that polycyclic aromatic hydrocarbons (PAHs) and alkylated PAHs, such as 1-and 3-methylchrysene, are important contributors to the overall estrogenic activity. However, these results revealed that a considerable proportion of the estrogenic activity in the soil remained unexplained, indicating the presence of other significant estrogenic compounds.

  • 48.
    Larsson, Maria
    et al.
    Örebro University, School of Science and Technology.
    Giesy, John P.
    Department of Veterinary Biomedical Sciences and Toxicological Center, University of Saskatchewan, Saskatoon, Saskatchewan, Canada; Department of Zoology and Center for Integrative Toxicology, Michigan State University, East Lansing, MI, USA; Department of Biology and Chemistry, State Key Laboratory in Marine Pollution, City University of Hong Kong, Kowloon, Hong Kong, China; School of Biological Sciences, University of Hong Kong, Hong Kong, China; State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing, People's Republic of China.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    AhR-mediated activities of polycyclic aromatic compound (PAC) mixtures are predictable by the concept of concentration addition2014In: Environment International, ISSN 0160-4120, E-ISSN 1873-6750, Vol. 73, p. 94-103Article in journal (Refereed)
    Abstract [en]

    Risk assessments of polycyclic aromatic hydrocarbons (PAHs) are complicated because these compounds exist in the environment as complex mixtures of hundreds of individual PAHs and other related polycyclic aromatic compounds (PACs). In this study, the hypothesis that concentration addition (CA) can be used to predict the aryl hydrocarbon receptor (AhR)-mediated activity of PACs in mixtures containing various combinations of PACs was tested. AhR-mediated activities of 18 mixtures composed of two to 23 PACs, which included PAHs, azaarenes and oxygenated PAHs, were examined by the use of the AhR-based H4IIE-luc bioassay. Since greater AhR-mediated activities have been observed in soils contaminated by PAHs, investigations were done to test whether soil extract matrix or the presence of non-effect PACs might affect responses of the H4IIE-luc bioassay. Our results showed that AhR-mediated activities of mixtures of PACs could be predicted by the use of concentration addition. Additive activities of PACs in multi component mixtures along with the insignificant effect of the soil matrix support the use of concentration addition in mass balance calculations and AhR-based bioassays in risk assessment of environmental samples. However, independent action (IA) could not be used to predict the activity of mixtures of PACs.

  • 49.
    Larsson, Maria
    et al.
    Örebro University, School of Science and Technology.
    Giesy, John P.
    Department of Veterinary Biomedical Sciences & Toxicological Center, University of Saskatchewan, Saskatoon, Canada; Department of Zoology, and Center for Integrative Toxicology, Michigan State University, USA Department of Biology & Chemistry and State Key Laboratory in Marine Pollution, City University of Hong Kong, China; State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, China.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Concentration-addition in risk assessment: prediction of potential AhR-mediated activity in multiple polycyclic aromatic compound (PAC) mixturesManuscript (preprint) (Other academic)
    Abstract [en]

    Risk assessments of polycyclic aromatic hydrocarbons (PAHs) are complicated because these compounds exist in the environment as complex mixtures of hundreds of individual PAHs and other related polycyclic aromatic compounds (PACs). In this study, the hypothesis that concentration addition (CA) can be used to predict the aryl hydrocarbon receptor (AhR) mediated potency of PACs in mixtures containing various combinations of PACs. Effects of 18 mixtures composed of two to 23 PACs, which included PAHs, azaarenes and oxygenated PAHs, were examined by use of the AhR based H4IIE-luc bioassay. Since greater AhR-mediated activities have been observed in soils contaminated by PAH, investigations were done to test whether soil extract matrix or presence of noneffect PACs might affect responses of the H4IIE-luc bioassay. AhR-mediated activity of the mixture of PACs could be predicted by use of concentration addition. Additive behavior of PACs in multi component mixtures supported the hypothesis that use of concentration addition could be used in risk assessment of PAC- mixtures. However, independent action (IA) could not be used to predict the activity of mixtures of PACs. 

  • 50.
    Larsson, Maria
    et al.
    Örebro University, School of Science and Technology.
    Hagberg, Jessika
    Örebro University, School of Science and Technology.
    Giesy, John P.
    Univ Saskatchewan, Dept Vet Biomed Sci, Saskatoon, SK, Canada; Univ Saskatchewan, Toxicol Ctr, Saskatoon, SK, Canada; Michigan State Univ, Dept Zool, E Lansing, USA; Michigan State Univ, Ctr Integrat Toxicol, E Lansing, USA; City Univ Hong Kong, Dept Biol & Chem, Kowloon, Hong Kong, Peoples R China; City Univ Hong Kong, State Key Lab Marine Pollut, Kowloon, Hong Kong, Peoples R China; Univ Hong Kong, Sch Biol Sci, Hong Kong, Hong Kong, Peoples R China; Nanjing Univ, Sch Environm, State Key Lab Pollut Control & Resource Reuse, Nanjing 210008, Jiangsu, Peoples R China .
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Time-dependent relative potency factors for polycyclic aromatic hydrocarbons and their derivatives in the H4IIE-luc bioassay2014In: Environmental Toxicology and Chemistry, ISSN 0730-7268, E-ISSN 1552-8618, Vol. 33, no 4, p. 943-953Article in journal (Refereed)
    Abstract [en]

    The H4IIE-luc transactivation bioassay for aryl hydrocarbon receptor (AhR) agonists was used to investigate the relative potency factors (REPs) of 22 individual polycyclic aromatic hydrocarbons (PAHs) and their oxygenated-, methylated-, and N-containing derivatives (azaarenes), which are often present in PAH-contaminated soils. Naphthacene and dibenz[ah]acridine exhibited greater AhR-mediated potency, whereas lesser molecular-weight azaarenes were less potent AhR agonists. Six oxygenated PAHs had calculable REPs, but their potencies were less than their parent PAHs. Unlike the parent, unsubstituted PAHs, oxidation of methylated PAHs seemed to increase the AhR-mediated potency of the compounds, with 2-methylanthracene-9,10-dione being almost 2 times more potent than 2-methylanthracene. Both bioassay and gas chromatography-mass spectrometry analysis were used to examine the exposure time-dependent effects on the REPs at 24 h, 48 h, and 72 h of exposure in the H4IIE-luc transactivation bioassay. Changes in concentrations of 5 compounds including the model reference 2,3,7,8-tetrachlorodibenzo-p-dioxin in the cell culture wells were measured, and the amounts in the cell medium, in the cells, and adsorbed to the wells was determined and the influence on the REPs was studied. Declining REP values with increased duration of exposure were shown for all compounds, which we concluded were a consequence of the metabolism of PAHs and PAH derivatives in H4IIe-luc cells. The present study provides new knowledge regarding the degradation and distribution of compounds in the wells during exposure. Environ Toxicol Chem 2014;33:943-953. (c) 2014 SETAC

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