Effect-directed fractionation and identification of cytochrome P4501A-inducing halogenated aromatic hydrocarbons in a contaminated sediment (original) (raw)
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Water Research, 1993
The activity of a P-450-dependent enzyme, ethoxyresorufin-O-deethylase (EROD), was measured in liver of fish from a watershed exposed to pesticide runoff from nearby vineyards. A correlation was observed between pesticide runoff and enzyme activity. This is the first field trial on the use of the cytochrome P-450 system as a biochemical indicator for assessing potential exposure to trace levels of complex mixtures of pesticides in the aquatic environment. The results might be useful for agencies designing monitoring assessment programs for ecotoxicological risk.
Marine Environmental Research, 1999
In this study, the degree of induction of cytochrome P4501A-associated 7-ethoxyresoru®n O-deethylase (EROD) activity and immunochemical detection of cytochrome P4501A in leaping mullet (Liza saliens) and common sole (Solea vulgaris) were used as biomarker for assessment of polycyclic aromatic hydrocarbon (PAH) or/and polychlorinated biphenyl (PCB) type organic pollutants along the Izmir Bay on the Aegean Sea coast, Turkey. Fish were captured in May 1995 and February and June 1996. Mullet caught from Pasaport, a highly urbanized and industrial section of the Bay, had approximately 62 times more EROD activity than the feral ®sh sampled from an uncontaminated site in the Outer Bay. Mullet caught along the pollutant gradient at the three other sites in the Bay exhibited less but highly signi®cant induced EROD activity. An inverse relationship was found between the EROD activity in the ®sh and the distance between the catch point and the discharge region of polluted rivers and of industrial and domestic wastes into the Harbour. Studies using the polyclonal antibodies produced against mullet cytochrome P4501A showed a similar trend. In addition, EROD activities of benthic ®sh, common sole, captured from three dierent sites of the Bay also con®rmed and extended the results obtained with those of mullet. Except that, common sole caught from site 5, the edge of Gediz river, exhibited higher EROD activity than that of ®sh caught from site 4, Tuzla. Although, detailed qualitative and quantitative analyses of organic Marine Environmental Research 48 (1999) 147±160 www.elsevier.com/locate/marenvrev 0141-1136/99/$ -see front matter # 1999 Published by Elsevier Science Ltd. All rights reserved. P I I : S 0 1 4 1 -1 1 3 6 ( 9 9 ) 0 0 0 3 8 -0 $ Preliminary results of this work have been presented in NATO-ASI on Molecular and Applied chemicals in the waters and sediments of Izmir Bay are not available, the results of this study indicate that Inner and Middle Bays of Izmir Bay are highly contaminated with PAH and/or PCB type organic pollutants. #
Toxicological sciences : an official journal of the Society of Toxicology, 2005
The major aim of this study was to characterize toxic organic compounds in bottom sediments from a PCB polluted bay. To overcome difficulties in pinpointing toxicants in complex environmental samples we applied a bio-effect directed (BED) fractionation approach and investigated the relationships between aromaticity, teratogenicity, and aryl hydrocarbon receptor (AhR) mediated toxicity. Hepatic ethoxyresorufin O-deethylase (EROD) activities and malformations were investigated in rainbow trout (Oncorhynchus mykiss) larvae exposed by injecting sediment extract and fractions (separated by their degree of aromaticity) thereof into newly fertilized eggs. Our results imply that non-additive effects get more pronounced the more complex the exposure. The fraction mainly composed of dicyclic aromatic compounds (DACs), including PCBs, was surprisingly less teratogenic than the fraction mainly composed of polycyclic aromatic compounds (PACs). A major part of the latter potential was isolated in...
Ecotoxicology and Environmental Safety, 1999
Along with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 24 unsubstituted polycyclic aromatic hydrocarbons (PAHs) were evaluated for their ability to induce 7-ethoxyresoru5n-odeethylase (EROD) activity in the rainbow trout liver cell line RTL-W1. When the duration and cell density of exposure were increased, the EC 50 for EROD induction was relatively constant for TCDD, but increased for PAHs. Regardless of exposure conditions, EROD activity was not induced by 9 PAHs: naphthalene, phenanthrene, anthracene, pyrene, perylene, acenaphthylene, acenaphthene, 6uorene, and 6uoranthene. Two PAHs, benzo[g,h,i]perylene and coronene, induced EROD activity inconsistently. The remaining 13 PAHs consistently induced EROD activity. The EC 50 s for induction exhibited approximately a 110-fold range. The order of potency, from most to least potent, was benzo[k]6uoranthene, dibenzo[a, i]pyrene, dibenzo [a,h]anthracene, benzo[a]pyrene, indeno[1,2,3-cd]pyrene, benzo [b]6uoranthene, pentacene, benzo[b]anthracene, benzo[b]
Environmental Toxicology and Chemistry, 1993
Mirror carp were exposed to Rotterdam Harbor sediment, highly contaminated with polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs), and polychlorobiphenyls (PCBs) (0.5 pg 2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD] equivalents per kilogram dry weight). In two additional separate experiments rainbow trout (Oncorhynchus mykiss) and mirror carp (Cyprinus carpio) received a single intraperitoneal injection of approximately 0.01,0.03, 0.06,0.3, 0.6, or 3.0 pg TCDD per kilogram body weight. In all three experiments induction of hepatic P450 1A was measured with immunochemical and enzymatic methods. The polyclonal antibodies anticod (Gadus morhua), anti-perch (Percafluviatilis), and anti-rainbow trout P450 1A all cross-reacted with the P450 1A orthologue of the carp and rainbow trout. In most cases high correlations were found between 7-ethoxyresorufin 0-deethylation (EROD) activity and cytochrome P450 1A protein contents, the latter measured by the enzyme-linked immunosorbent assay (ELISA) and protein blot methods. However, the correlations between EROD activity and P450 1A protein levels were higher within the separate sampling periods (i.e., 3,6, and 12 weeks after dosage) than with the total data set, especially in the dose-effect study with the rainbow trout. This was probably caused by a difference in time-dependent relationships between P450 1A protein content and EROD enzyme activity: 12 weeks after dosage the P450 1A protein was still increased, although EROD activity had returned to background level. In addition, there were higher correlations of the EROD activity and P450 1A protein content with total P450 content in rainbow trout and carp treated with a single dose of TCDD, than with total P450 content in carp exposed to contaminated sediment. In our study, the ELISA method appeared to be more useful than the protein blot technique, because the ELISA is faster and has higher reproducibility. In addition, in all our experiments EROD activity showed a higher induction than the P450 1A protein, indicating a higher sensitivity of the EROD assay. Our results strongly indicated that determination of the P450 1A protein content and EROD activity provides complementary information. Thus we recommended the use of both the ELISA and the EROD activity assay in order to understand the nature of P450 1A induction.
Environment International, 2008
Sediment samples from the upper Danube River in Germany have previously been characterized as ecotoxicologically hazardous and contaminants in these sediments may contribute to the observed decline of fish populations in this river section. For the investigation of sediment toxicity there is a need for development, standardization and implementation of in vivo test systems using vertebrates. Therefore, the main objective of this study was to apply and evaluate a recently established fish gill EROD assay as a biomarker in sediment toxicity assessment by using extracts of well characterised sediment samples from the upper Danube River. This to our knowledge is the first application of this novel assay to sediment extracts. Sediments from four different sites along the upper Danube River were Soxhlet-extracted with acetone and dissolved in DMSO. Three-spined sticklebacks (Gasterosteus aculeatus L.) were exposed for 48 h to various concentrations of the extracts, to the positive control β-naphthoflavone or to the solvent. Measurements of EROD activity in gill filaments and liver microsomes followed the exposure. Concentrationdependent induction of EROD in both gill and liver was found for all sediment extracts. The highest ERODinducing potency was determined for extracts of sediments from the sites "Öpfinger See" and "Sigmaringen" and the EROD activities in gill and liver correlated well. The results from the gill and liver assays were in accordance with in vitro results of previous investigations. The EROD activities measured in the present study corresponded with the concentrations of PAHs, PCBs and PCDD/Fs in the sediment samples derived in a previous study. The sticklebacks in this study were in the reproductive phase and a stronger EROD induction was obtained in the females than in the males. Implementation of the EROD assay in testing of sediment extracts gave highly reliable results which make this assay an ecotoxicologically relevant method for assessment of contamination with Ah receptor agonists in sediments.
Environment International, 2008
Sediment samples from the upper Danube River in Germany have previously been characterized as ecotoxicologically hazardous and contaminants in these sediments may contribute to the observed decline of fish populations in this river section. For the investigation of sediment toxicity there is a need for development, standardization and implementation of in vivo test systems using vertebrates. Therefore, the main objective of this study was to apply and evaluate a recently established fish gill EROD assay as a biomarker in sediment toxicity assessment by using extracts of well characterised sediment samples from the upper Danube River. This to our knowledge is the first application of this novel assay to sediment extracts. Sediments from four different sites along the upper Danube River were Soxhlet-extracted with acetone and dissolved in DMSO. Three-spined sticklebacks (Gasterosteus aculeatus L.) were exposed for 48 h to various concentrations of the extracts, to the positive control β-naphthoflavone or to the solvent. Measurements of EROD activity in gill filaments and liver microsomes followed the exposure. Concentrationdependent induction of EROD in both gill and liver was found for all sediment extracts. The highest ERODinducing potency was determined for extracts of sediments from the sites "Öpfinger See" and "Sigmaringen" and the EROD activities in gill and liver correlated well. The results from the gill and liver assays were in accordance with in vitro results of previous investigations. The EROD activities measured in the present study corresponded with the concentrations of PAHs, PCBs and PCDD/Fs in the sediment samples derived in a previous study. The sticklebacks in this study were in the reproductive phase and a stronger EROD induction was obtained in the females than in the males. Implementation of the EROD assay in testing of sediment extracts gave highly reliable results which make this assay an ecotoxicologically relevant method for assessment of contamination with Ah receptor agonists in sediments.