Abstract
Thirteen individual organochlorine compounds at 3 concentrations (80, 400, and 2000 ng/mL culture medium), as well as mixtures, were assayed for the estrogen receptor (ER) activation or inhibition, using a luciferase reporter gene assay (RGA). None of the PCB 138, 153, or 180 or their mixture induced a response in the RGA. o,p′-DDT was the most potent xenoestrogen from the DDT group, inducing a response already at 80 ng/mL. From the HCH and HCB group, only β-HCH (at 400 and 2000 ng/mL) and δ-HCH (at 2000 ng/mL) displayed estrogenic activities. These 13 organochlorines were determined by GC-MS in 12 samples of North Sea harbor porpoise blubber. The PCBs were the main contaminants. Within each group, PCB 153 (6.0 × 102~4.2 × 104 μg/kg), p,p′-DDE (5.1 × 102~8.6 × 103 μg/kg), and HCB (7.6 × 101~1.5 × 103 μg/kg) were the compounds found in highest concentrations. The hormonal activity of the porpoise blubber samples was also assayed in RGA, where two samples showed estrogenic activity, seven samples showed antiestrogenic activity, and one sample showed both estrogenic and antiestrogenic activity. Our results suggest that the 13 POPs measured by GC-MS in the samples cannot explain alone the estrogenicity of the extracts.
Highlights
The harbour porpoise (Phocoena phocoena) is the most common cetacean species in the North Sea [1], and there is a growing concern about the adverse effects of persistent environmental contaminants on this and other marine mammal species [2]
The following equipment and materials have been used in this study: carbon dioxide (Air Liquide, Liege, Belgium); helium (Air Products, Brussels, Belgium); acetonitrile (Biosolve, Valkenswaard, Netherlands); luminometer Orion II (BRS, Drogenbos, Belgium); adenosine triphosphate (ATP), Dulbecco’s modified Eagle’s medium (DMEM), DMEM without red phenol, and fetal bovine serum and trypsin (Fisher Bioblock Scientific, Tournai, Belgium); Focus gas chromatograph and Polaris Q mass spectrometer (Interscience, Louvain-laNeuve, Belgium); charcoal, dextran, o,p-DDD, p,p-DDD, p,p-DDE, o,p-DDT, p,p-DDT, 17β-estradiol, HCB, α-HCH, β-HCH, γ-HCH, δ-HCH, nonane, PCB 80 13C, PCB 138 (2,2,3,4,4,5-hexachlorobiphenyl), PCB 153 (2,2,4,4,5,5hexachlorobiphenyl), and PCB 180 (2,2,3,4,4,5,5-heptachlorobiphenyl) (Sigma-Aldrich, Bornem, Belgium); Dluciferin (Synchem OHG, Kassel, Germany); HT8 column (VWR, Leuven, Belgium)
The study of estrogen receptor- (ER-) mediated activity of persistent organic pollutants (POPs) in MCF-7-ERE cells showed that several organochlorine pollutants (DDT and metabolites and HCH isomers) present an estrogenic activity, which was confirmed by other authors [24, 44,45,46,47]
Summary
The harbour porpoise (Phocoena phocoena) is the most common cetacean species in the North Sea [1], and there is a growing concern about the adverse effects of persistent environmental contaminants on this and other marine mammal species [2]. Since 1998, the southern region of the North Sea has been characterised by an increased number of stranded marine mammals, in particular the harbour porpoise. A temporary increase in the porpoise population in the southern North Sea may have been responsible [3, 4]. Since marine mammals are at the top of the aquatic food chain and have rather long lifespans, they are an important tool to check the long-term effects concerning marine environment pollution and can be studied as global pollution indicators as well [5].
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