Monitoring river sediments contaminated predominantly with polyaromatic hydrocarbons by chemical and in vitro bioassay techniques

Abstract

Extracts of sediment samples collected from the Morava River and its tributaries (Czech Republic) were examined for mutagenic, dioxin-like, and estrogenic activities. Moreover, the human leukemic HL-60 cell line was tested as a potential model for the detection of effects of environmental contaminants on cell proliferation and differentiation processes. Analytical data indicate that the sediments were contaminated predominantly with polycyclic aromatic hydrocarbons (PAHs) and phthalate esters. The sums of concentrations of 16 U.S. Environmental Protection Agency priority PAHs ranged from 0.8 to 13.2 micrograms/g and those of phthalates reached up to 3,000 ng/g, while only low levels of chlorinated hydrocarbons were found. The main goal of the present study was to determine effects of PAH prevalence on in vitro bioassays, with special emphasis on dioxin-like activity. The dioxin-like activity was tested using a reporter gene assay based on chemical-activated luciferase expression (the CALUX assay). Significant dioxin-like activity (2.6-40.1 micrograms/g benzo[a]pyrene equivalents and 5.9-48.2 ng/g 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents) was detected in all samples, and the results obtained with various exposure times or with both crude and PAH-deprived extracts indicate that the response was probably caused almost exclusively by the presence of high concentrations of PAHs. This corresponds with results of chemical analyses and indicates that various exposure times would allow a discrimination between dioxin-like activities of persistent compounds and easily metabolized aryl hydrocarbon (Ah) receptor inducers. Only sediment extracts containing the highest concentrations of PAHs were mutagenic, as determined by the umu assay. Estrogenic activity was found in several samples (4.75-22.61 pg/g estradiol equivalents) using cells stably transfected with an estrogen-responsive element linked to a luciferase promoter. Noncytotoxic doses of extracts had no effects on HL-60 cell proliferation, while two of the tested crude extracts significantly enhanced their all-trans retinoic acid-induced differentiation. These activities were not associated with phthalate esters and/or PAHs. Our results indicate that cellular and biochemical in vitro assays based on various specific modes of action may yield data complementary to results of mutagenicity tests and that they could be useful in environmental risk assessment. High levels of PAHs are apparently associated with dioxin-like and mutagenic activities rather than with estrogenic activity.

Polycyclic aromatic hydrocarbons in surface sediments of the Jialu River

“…The PAH concentrations in surface sediments from the Jialu River in this study are similar to those detected in the River Gomti, India (Tripathi et al 2009) and the Yellow River, China (Xu et al 2007), and higher than those found at the Yalujiang River, China (Wu et al 2003), the Sava River, Serbia (Crnković et al 2008), the Luan River, China (Bai et al 2008), the Daliao River, China and the Gao-Ping River, Taiwan (Doong and Lin 2004). However, the levels were lower than those reported in the Zhujiang River, China (Mai et al 2002), the Passaic River, USA (Huntley et al 1995) and the Morava River, Czech Republic (Vondracek et al 2001).…”

Contamination of riverbed sediments by hazardous substances in the Mediterranean context: Influence of hydrological conditions

“…Except in March 2010, when the peak PAH concentration was recorded, PAH concentrations measured in the sediments of the Vène river were in the same range as concentrations observed in other French Mediterranean rivers (Table 4). Although this high PAH concentration (10,188 ng/g dw) was lower than the concentrations of up to 23,078 ng/g dw measured in rivers in dense urban and industrial areas in France (Kinani et al, 2010), this PAH concentration in the Vène river was in the range of concentrations observed in industrial areas: 636-13,205 ng/g dw (Vondracek et al, 2001) and 5.3-7700 ng/g dw (Koh et al, 2004) and could consequently be toxic for organisms.…”

Wirkungsorientierte Analytik komplexer Mischungen Erfahrungen aus der Umwelt- und Ökotoxikologie für die Lebensmitteluntersuchung

“…Sie zeichnet sich durch sehr hohe Trennschärfe und sehr gute Reproduzierbarkeit aus, was eine automatisierte Vielfachinjektion ermöglicht und damit das Problem der geringeren isolierten Stoffmenge pro Injektion ausgleicht. (Ames et al, 1975;Samoiloff et al, 1983;DeMarini et al, 1994) Ames Fluktuationstest in Flüssigkultur (Mikrotiterplatten) (Pérez et al, 2003;Flückiger-Isler et al, 2004) Mutatox: Rückmutation einer Dunkelmutante von Vibrio fischeri (Ho and Quinn, 1993;Thomas et al, 2002a) Mutagenitätstests mit menschlichen h1A1v2 B-Lymphoblastoid-Zellen mit constitutiver Überexpression der Oxygenase P4501A1 (Durant et al, 1998;Hannigan et al, 1998) Ah-Rezeptor-vermittelte Wirkung Induktion von 7-Ethoxyresorufin-O-Deethylase in Leberzelllinien von Fischen (RTL-W1, PLHC-1) und Ratten (H4IIE) (Safe, 1986;Brack et al 2002;Gale et al, 2000;Hilscherova et al, 2001) Luziferase-Aktivität in rekombinanter Zelllinie H4IIE-luc (CALUX) Vondracek et al, 2001) Estrogene Aktivität Zelltest mit menschlichen Brustkrebszellen (MCF-7) Zelltest mit menschlichen Brustkrebszellen mit rekombinantem RezeptorReporter-Gen (MCF-7-luc oder MVLN) (Soto et al, 1995) (Vondracek et al, 2001 Hefe-basierter Zelltest mit recombinantem Östrogen-Rezeptor-Reporter-Gen (YES) (Thomas et al, 2004) Androgene Aktivität Hefe-basierter Zelltest mit rekombinantem Androgen-Rezeptor-Reporter-Gen (Thomas et al, 2004;Thomas et al, 2002b) Zellproliferation und Differenzierung…”

“…Menschliche Leukämiezellen (HL-60) oder Rattenleberepitheliumzellen (WB-F344) (Vondracek et al, 2001;Vondracek et al, 2004) GJIC-Hemmung Hemmung der interzellulären Kommunikation bei WB-F344 und Aufhebung der Wachstumshemmung durch Zellkontakt (Tumorpromotion) (Vondracek et al, 2004) Immuntoxizität Hemmung der Lymphoid-Entwicklung in Thymuszellen von Mausföten (C57BL/6) Phagozytosetest Bursttest (Engwall et al, 1997) Entzündungsförderung Cytokin-Produktion in peripheren mononukleären Blutzellen (Wichmann et al, 2004) Prinzipiell sind alle in der Umweltforschung eingesetzten Fraktionierungsverfahren für Lebensmittelextrakte geeignet.…”