Monitoring river sediments contaminated predominantly with polyaromatic hydrocarbons by chemical and in vitro bioassay techniques.
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.
Vondrácek
J
J
Institute of Biophysics, Czech Academy of Sciences, 61265 Brno, Czech Republic, The Netherlands.
Machala
M
M
Minksová
K
K
Bláha
L
L
Murk
A J
AJ
Kozubík
A
A
Hofmanová
J
J
Hilscherová
K
K
Ulrich
R
R
Ciganek
M
M
Neca
J
J
Svrcková
D
D
Holoubek
I
I
D001681:Biological Assay; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D004784:Environmental Monitoring / Q000379:methods*; D004785:Environmental Pollutants / Q000009:adverse effects / Q000032:analysis*; D005786:Gene Expression Regulation / Q000187:drug effects; D019015:Geologic Sediments / Q000737:chemistry; D018922:HL-60 Cells; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D011084:Polycyclic Aromatic Hydrocarbons / Q000009:adverse effects / Q000032:analysis*; D018570:Risk Assessment; D018675:Toxicity Tests; D014874:Water Pollutants, Chemical / Q000032:analysis*
D004785:Environmental Pollutants; D011084:Polycyclic Aromatic Hydrocarbons; D014874:Water Pollutants, Chemical; D008156:Luciferases