Effect‐directed analysis of mutagens and ethoxyresorufin‐O‐deethylase inducers in aquatic sediments

Sediment extracts from a creek in the Neckar river basin (Germany), which received the discharge of treated hospital wastewater, were found to exhibit strong aromatic hydrocarbon (Ah) receptor-mediated effects in a rainbow trout liver cell line (RTL-WI) as well as high mutagenicity in the Salmonella/microsome assay after fractionation. The crude extract did not exhibit a clear mutagenic response. Apparently, cleanup or fractionation before mutagenicity testing is necessary to minimize the risk of false-negative results. Effect-directed fractionation and analysis were applied to characterize and identify the toxicants that cause these effects. Major ethoxyresorufin-O-deethylase induction potency and mutagenicity were detected in different polyaromatic fractions, indicating different sets of toxicants that induce metabolic activation and mutagenicity. Dioxin-like halogenated aromatic hydrocarbons, including polychlorinated biphenyls, naphthalenes, dibenzo-p-dioxins and furans, and priority polycyclic aromatic hydrocarbons, contributed to Ah receptor-mediated activity only to a minor extent. Benzo[a]pyrene, benzo[a]fluoranthene, and perylene could be confirmed as important contributors to mutagenicity. The nonpriority pollutants 11H-indeno[2,1,7-cde]pyrene, a methylbenzo[e]pyrene, and a methylperylene were tentatively identified as major components, representing 82% of the peak area of a highly mutagenic fraction of the sediment extract. This suggests that hazard and risk assessment of complex environmental mixtures should make increasing attempts to identify and consider hazardous key pollutants rather than focusing on a priori-selected key pollutants alone.

[1]  F. Guengerich,et al.  Oxidation of benzo[a]pyrene by recombinant human cytochrome P450 enzymes. , 1995, Chemical research in toxicology.

[2]  T. Braunbeck,et al.  Cytotoxicity of settling particulate matter and sediments of the Neckar River (Germany) during a winter flood , 2000 .

[3]  J. Gustafsson,et al.  Short-term bioassays of nitro derivatives of benzo[a]pyrene and perylene. , 1984, Carcinogenesis.

[4]  L. Johnson,et al.  Overview of studies on liver carcinogenesis in English sole from Puget Sound; evidence for a xenobiotic chemical etiology. I: Pathology and epizootiology. , 1990, The Science of the total environment.

[5]  M. M. Krahn,et al.  Relationships between hepatic neoplasms and related lesions and exposure to toxic chemicals in marine fish from the U.S. West Coast. , 1991, Environmental health perspectives.

[6]  P. White The genotoxicity of priority polycyclic aromatic hydrocarbons in complex mixtures. , 2002, Mutation research.

[7]  Douglas S. Galvao,et al.  Identifying carcinogenic activity of methylated polycyclic aromatic hydrocarbons (PAHs) , 1999 .

[8]  C. Ioannides,et al.  Interaction with the aromatic hydrocarbon receptor, CYP1A induction, and mutagenicity of a series of diaminotoluenes: implications for their carcinogenicity. , 1996, Toxicology and applied pharmacology.

[9]  J. Vondráček,et al.  Aryl hydrocarbon receptor-mediated activity of mutagenic polycyclic aromatic hydrocarbons determined using in vitro reporter gene assay. , 2001, Mutation research.

[10]  G. Grimmer Analysis of automobile exhaust condensates. , 1977, IARC scientific publications.

[11]  H. Bende Polycyclische aromatische Kohlenwasserstoffe , 1967 .

[12]  Wolfgang Ahlf,et al.  Application of a Sediment Quality Triad and Different Statistical Approaches (Hasse Diagrams and Fuzzy Logic) for the Comparative Evaluation of Small Streams , 2002, Ecotoxicology.

[13]  T. Rushmore,et al.  Pathogenesis of skin and liver neoplasms in white suckers from industrially polluted areas in Lake Ontario. , 1990, The Science of the total environment.

[14]  K. Schirmer,et al.  Cell and Tissue Culture , 2000 .

[15]  J. Bayona,et al.  Characterization of genotoxic components in sediments by mass spectrometric techniques combined withSalmonella/microsome test , 1990, Archives of environmental contamination and toxicology.

[16]  R. K. Saugier,et al.  The relationship between ionization potential and horseradish peroxidase/hydrogen peroxide-catalyzed binding of aromatic hydrocarbons to DNA. , 1983, Chemico-biological interactions.

[17]  A. Kozubík,et al.  Induction of aryl hydrocarbon receptor–mediated and estrogen receptor–mediated activities, and modulation of cell proliferation by dinaphthofurans , 2004, Environmental toxicology and chemistry.

[18]  W. Brack,et al.  Sequential fractionation procedure for the identification of potentially cytochrome P4501A-inducing compounds. , 2003, Journal of chromatography. A.

[19]  Mark S. Myers,et al.  Neoplastic and other diseases in fish in relation to toxic chemicals: an overview☆ , 1988 .

[20]  Ronald A. Hites,et al.  The global distribution of polycyclic aromatic hydrocarbons in recent sediments , 1978 .

[21]  P. Popp,et al.  Identification and quantification of thiaarenes in the flue gas of lignite-fired domestic heating , 2000 .

[22]  Helmut Segner,et al.  Fixed‐effect‐level toxicity equivalents—a suitable parameter for assessing ethoxyresorufin‐O‐deethylase induction potency in complex environmental samples , 2000 .

[23]  Miroslav Machala,et al.  Inhibition of gap-junctional intercellular communication by environmentally occurring polycyclic aromatic hydrocarbons. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[24]  T. Braunbeck,et al.  Changes in toxicity and genotoxicity of industrial sewage sludge samples containing nitro- and amino-aromatic compounds following treatment in bioreactors with different oxygen regimes , 2004, Environmental science and pollution research international.

[25]  W. Brack,et al.  Effect-directed identification of oxygen and sulfur heterocycles as major polycyclic aromatic cytochrome P4501A-inducers in a contaminated sediment. , 2003, Environmental science & technology.

[26]  B. Ames,et al.  Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. , 1975, Mutation research.

[27]  J. Westendorf,et al.  The use of the Salmonella BA9 forward mutation assay in sediment quality assessment: mutagenicity of freshly deposited sediments of the River Elbe , 1995 .

[28]  M. Costa,et al.  DNA methylation, heterochromatin and epigenetic carcinogens. , 1997, Mutation research.

[29]  H. Kruk,et al.  Electronic considerations in the mutagenesis of some 4,5-bridged chrysenes. , 1987, Environmental mutagenesis.

[30]  M. Tysklind,et al.  Biological and Chemical Determination of Dioxin-like Compounds in Sediments by Means of a Sediment Triad Approach in the Catchment Area of the River Neckar , 2002, Ecotoxicology.

[31]  E. Zeiger,et al.  Suppressive effects of chemicals in mixture on the Salmonella plate test response in the absence of apparent toxicity. , 1984, Environmental mutagenesis.

[32]  W. Manz,et al.  Comparative genotoxicity testing of rhine river sediment extracts using the comet assay with permanent fish cell lines (rtg-2 and rtl-w1) and the ames test* , 2004 .

[33]  S. J. Caldwell,et al.  Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity , 1993, Cell Biology and Toxicology.

[34]  Werner Brack,et al.  Effect‐directed fractionation and identification of cytochrome P4501A‐inducing halogenated aromatic hydrocarbons in a contaminated sediment , 2002, Environmental toxicology and chemistry.

[35]  J. Clark,et al.  Epizootiology of neoplasms in bony fish of North America. , 1990, The Science of the total environment.

[36]  B. Ames,et al.  Revised methods for the Salmonella mutagenicity test. , 1983, Mutation research.

[37]  Determination of genotoxic polycyclic aromatic hydrocarbons in a sediment from the Black River (Ohio) , 1986, Archives of environmental contamination and toxicology.

[38]  C. Metcalfe,et al.  Identification of potential fish carcinogens in sediment from hamilton harbour, ontario, canada , 1995 .

[39]  M. Servos,et al.  Mammalian and teleost cell line bioassay and chemically derived 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin equivalent concentrations in lake trout (Salvelinus namaycush) from Lake Superior and Lake Ontario, North America , 1998 .

[40]  D. Schaeffer,et al.  Quantifying the toxic and mutagenic activity of complex mixtures with Salmonella typhimurium. , 1981, Journal of toxicology and environmental health.

[41]  P. Hebert,et al.  Effects of chemical contaminants on genetic diversity in natural populations: implications for biomonitoring and ecotoxicology. , 2000, Mutation research.