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Eunate Puy-Azurmendi,
Maren Ortiz-Zarragoitia,
Marta Villagrasa,
Marina Kuster,
Pilar Aragón,
Julia Atienza,
Rosa Puchades,
Angel Maquieira,
Carmen Domínguez,
Miren López de Alda, Denise Fernandes,
Cinta Porte,
Josep M Bayona,
Damià Barceló,
Miren P Cajaraville
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ABSTRACT: Endocrine disruptors (EDs) interfere with the development and functioning of the endocrine system, causing reproductive disturbance in aquatic wildlife. The aim of the present work was to determine the presence of EDs in sediments and to investigate possible exposure and effects of EDs in the estuary of the Urdaibai Biosphere Reserve (Gernika) in comparison with the Arriluze marina. For this, gonad histology, plasma vitellogenin (VTG) protein levels and mRNA levels of vitellogenin (vtg), cyp19 aromatases, estrogen receptor (er) and retinoid X receptor (rxr) were studied in Chelon labrosus. The presence of alkylphenols (APs) in fish bile was also assessed. In sediments, estrogenic hormones were below the detection limit and levels of bisphenol A were very low. In Gernika organotin compounds were low but in Arriluze levels of up to 12μg/g were found. Moderate levels of APs and phthalate levels of up to 8μg/g were found in sediments. In fish, a high prevalence up to 33% of intersex gonads was found in Gernika, whereas only one intersex was found in Arriluze. Accordingly, mullets from Gernika showed higher concentrations of APs in bile. VTG protein levels were detected not only in females but also in some undifferentiated, male and intersex fish. mRNA of vtg was detected in one male from Gernika. mRNA of er and rxr showed significant differences between seasons. In conclusion, the present study demonstrated that C. labrosus from the Urdaibai estuary were exposed to EDs and showed clear signs of endocrine disruption.
Science of The Total Environment 11/2012; 443C:233-244. · 3.29 Impact Factor
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ABSTRACT: The in vitro interference of fibrate (gemfibrozil, clofibrate, clofibric acid), anti-inflammatory (ibuprofen, diclofenac), and anti-depressive (fluoxetine, fluvoxamine) drugs with key enzymatic activities-C17,20-lyase and CYP11β-involved in the synthesis of active androgens in gonads of male carp have been investigated. Among the tested compounds, fluvoxamine and fluoxetine were the strongest inhibitors of C17,20-lyase and CYP11β enzymes, with IC50s in the range of 321-335 μM and 244-550 μM, respectively. To our knowledge this is the first report on the interaction of pharmaceutical compounds with enzymatic systems involved in the synthesis of oxy-androgens. As oxy-androgens are known to influence spermatogenesis and stimulate reproductive behavior and secondary sexual characteristics in male fish, this work highlights the need for further investigating these endpoints when designing specific in vivo studies to assess the endocrine disruptive effect of pharmaceuticals in fish.
Marine pollution bulletin 08/2011; 62(10):2250-3. · 2.63 Impact Factor
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ABSTRACT: Molluscs are the second most diverse animal group, they are ecologically important and they are considered excellent indicators of ecosystem health. Some species have been widely used in pollution biomonitoring programs; however, their endocrinology is still poorly known. Despite some studies reporting the presence of (vertebrate-type) steroids in molluscs, information regarding enzymatic pathways involved in steroid synthesis and further catabolism of those steroids is still fragmentary. Regarding steroidogenesis, a number of excellent studies were performed in the 70s using different radio-labelled steroid precursors and detecting the formation of different metabolites. But, since then a long gap of research exist until the late 90s when the 'endocrine disruption' issue raised the need of a better knowledge of mollusc (and invertebrate) endocrinology in order to assess alterations caused by pollutants. Here we summarize past and recent studies dealing with steroid biosynthesis and metabolism in different mollusc species. Most of these studies suggest the involvement of steroids in mollusc reproduction. However, the knowledge is still fragmentary and many questions remain to be answered.
The Journal of steroid biochemistry and molecular biology 12/2010; 127(3-5):189-95. · 2.66 Impact Factor
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ABSTRACT: Mussels (Mytilus galloprovincialis) were exposed to different concentrations of testosterone (T: 20, 200 and 2000ng/L) in a semi-static water regime (1-day dosing intervals) for up to 5 days in an attempt to see whether endogenous steroid levels and steroid metabolism were altered by exogenous exposure to testosterone. Whole tissue levels of total testosterone (free+esterified) sharply increased in a concentration-dependent manner, from 2ng/g in controls to 290ng/g in organisms exposed to the highest concentration. In contrast, levels of free testosterone were only significantly elevated at the high-exposure group (5-fold increase with respect to controls). Increased activity of palmitoyl-CoA:testosterone acyltransferase (ATAT) was detected in organisms exposed to the highest concentration of testosterone, while those exposed to low and medium concentrations showed significant alterations in their polyunsaturated fatty acid profiles. The obtained results suggest that esterification of the excess of T with fatty acids might act as a homeostatic mechanism to maintain endogenous levels of free T stable. Interestingly, a decrease in CYP3A-like activity was detected in T-exposed mussels together with a significant decrease in the metabolism of the androgen precursor androstenedione to dihydrotestosterone (5α-DHT). Overall, the work contributes to the better knowledge of androgen metabolism in mussels.
Aquatic toxicology (Amsterdam, Netherlands) 11/2010; 100(4):313-20. · 3.12 Impact Factor
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ABSTRACT: The Urdaibai estuary is a UNESCO biosphere reserve impacted by recreational, agricultural and industrial activities. Polycyclic aromatic hydrocarbons (PAHs) are major pollutants in Urdaibai, and their control and the identification of their sources is central on the preservation of the area. Chemical analysis by GC-MS showed a significant pollution by PAHs in Urdaibai sediment samples, mainly from pyrolytic sources, with minor contributions from oil spills. Measurement of the dioxin-like activity using a yeast-based bioassay showed an excess of biological activity in sediment samples from the inner part of the estuary. Analysis of PAH metabolites in bile of thicklip grey mullet (Chelon labrosus) showed a large excess of 2-naphthol over 1-naphthol or 1-pyrenol, suggesting a specific contamination in the inner Urdaibai estuary by some industrial process. Therefore, the combination of these three techniques defined different PAH pollution sources in Urdaibai: a major pyrolytic origin, occasional oil spills, and specific industrial activities.
Marine environmental research 04/2010; 70(2):142-9. · 1.76 Impact Factor
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ABSTRACT: Synthetic musks are widely used as perfuming agents in products, such as cosmetics, detergents, and soaps. The increased detection of these substances in the aquatic environment and their high bioconcentration potential raises concerns about potential effects on aquatic species. This work aimed at assessing the interactions of the most widely used musks: nitromusks (musk xylene, musk ketone) and polycyclic musks (celestolide, galaxolide, and tonalide) with fish enzymatic systems involved in both xenobiotic and endogenous metabolism. Therefore, CYP catalyzed pathways were investigated in carp liver microsomes (CYP1A, CYP3A), ovarian microsomes (CYP19) and testicular mitochondria (CYP17 and CYP11beta) using standard substrates. Phase II activities (UDP-glucuronosyltransferases and sulfotransferases) were determined in carp liver microsomes and cytosol, respectively. Polycyclic musks (galaxolide and tonalide) were stronger inhibitors of CYP3A- (IC(50): 68-74 microM), CYP17- (IC(50): 213-225 microM), CYP11beta- and CYP19-catalyzed activities than nitromusks, while the latter showed higher ability to interfere with CYP1A (IC(50): 35-37 microM). The sulfation of estradiol was also significantly inhibited by tonalide and galoxolide (IC(50): 140-294 microM). Overall, polycyclic musks showed the highest potential to interfere with those activities involved in the synthesis and metabolism of steroids while nitromusks mainly interfered with xenobiotic metabolism (CYP1A-catalyzed reactions). The obtained data suggest that CYP isoforms are potentially sensitive targets of synthetic musk substances in fish.
Environmental Science and Technology 11/2009; 43(24):9458-64. · 5.23 Impact Factor
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ABSTRACT: The aquaculture industry is an expanding activity, mainly due to the world population demand of fish. However, intensive production and the use of chemicals have raised environmental concerns and have questioned the quality of cultured fish in comparison to wild fish. Up to date, there is little information regarding pollutant residues in cultured stocks and the risks associated to consumption. Here we summarize recent data on pollutants exposure, together with biochemical responses in both cultured and wild sea bass (Dicentrarchus labrax) from the Iberian Peninsula. The obtained data highlights the use of chemicals in current aquaculture practices as a significant source of pollution in cultured fish that poses a risk to exposed individuals and may negatively impact aquatic ecosystems. Nonetheless, a controlled use of those chemicals and a careful selection of the aquaculture location can ensure relatively low and homogeneous levels of pollutants in cultured fish in comparison to wild specimens.
Ecotoxicology 08/2009; 18(8):1043-50. · 2.36 Impact Factor
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ABSTRACT: There is little information regarding pollutant levels in farmed fish, and the risks associated to consumption. This study was designed to assess levels of exposure to metals, organochlorinated compounds, polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APEs) in farmed sea bass Dicentrarchus labrax from five aquacultures located in Southern Europe. Additionally, several biochemical responses (metallothionein, 7-ethoxyresorufin O-deethylase, vitellogenin) were determined as complementary tools. The obtained data indicate that pollutants exposure in farmed fish is similar to the levels reported in wild specimens from the area. Nonetheless, some biochemical responses were observed in the studied organisms, viz. metallothionein induction in Cu exposed organisms, and 7-ethoxyresorufin O-deethylase (EROD) and vitellogenin induction in PAHs and APEs exposed ones. The study further supports the usefulness of the biomarker approach as a first screening method to discriminate between basal and high levels of exposure in cultured fish.
Environmental Pollution 04/2008; 152(1):138-46. · 3.75 Impact Factor
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ABSTRACT: Metal levels (Cd, Pb, Hg, Cr, Fe, Cu, Zn) together with metallothioneins (MTs) were determined in the liver of two commercial fish species collected along the Northern Iberian coast. The four-spotted megrim (Lepidorhumbos boscii) and the pouting (Trisopterus luscus) were selected as representative species of the middle/outer (200-500 m) and inner shelf (70-120 m), respectively. Metal pollution in the middle/outer shelf was mostly detected in the Asturias and Basque Country areas, whereas in the inner shelf, pollution was widely spread along the coast. Significantly high levels of Pb, Cd, Cr and Hg were detected in T. luscus from different sampling sites and associated to anthropogenic pressure. MTs were induced in fish that had higher amounts of Zn, Cd, Cr and Hg in their livers, supporting the use of MT as a biomarker of metal exposure. The study supports the usefulness of the selected fish species as sentinel organisms for future biomonitoring studies.
Science of The Total Environment 03/2008; 391(1):159-67. · 3.29 Impact Factor
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ABSTRACT: The metabolism of 17alpha-hydroxyprogesterone (17P(4)) was investigated in different subcellular fractions isolated from male gonads of sea bass (Dicentrarchus labrax L). The existence of CYP17 (C17,20-lyase activity) and CYP11B (11beta-hydroxylase) catalyzed reactions was demonstrated in the mitochondrial fraction, where 17P(4) was converted to androstenedione (AD) and further metabolized to 11beta-hydroxyandrostenedione (betaAD). The synthesis of betaAD predominated in early spermatogenic testis, indicating a role of betaAD in testicular recrudescence. Additionally, the in vitro effect of model endocrine disrupting chemicals (i.e. nonylphenol (NP), p,p'-DDE, benzo[a]anthracene (BaA), tributyltin (TBT) and ketoconazole (KCZ)) on the mitochondrial metabolism of 17P(4) was investigated. Among the tested compounds, 100 microM NP inhibited the activity of CYP17 (C17,20-lyase) whereas 100 microM KCZ inhibited both CYP17 and CYP11B. Both chemicals showed the potential to disrupt the reproductive cycle of fish living in polluted environments due to impairment of testicular steroid biosynthesis. These results suggest that mitochondrial metabolism of 17P(4) may constitute a new sensitive probe for the assessment of endocrine disruption in fish.
Aquatic Toxicology 01/2008; 85(4):258-66. · 3.76 Impact Factor
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ABSTRACT: Muscle concentrations of organochlorinated compounds as well as biliary levels of polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APEs) were determined in two different fish species, the four-spotted megrim (Lepidorhombus boscii) and the pouting (Trisopterus luscus) collected along the Northern Iberian coast. Additionally, a set of biochemical markers namely, 7-ethoxyresorufin O-deethylase (EROD), UDP-glucuronosyltransferase (UGT) and catalase (CAT) were measured in liver subcellular fractions. Chemical analysis indicated geographical differences in pollutant loads that were further reinforced by biomarker responses. Thus, EROD activity showed a good correlation with the amount of PCBs bioaccumulated in muscle tissue of both fish species. Elevated UGT activity was observed in those individuals highly exposed to APEs and 1-naphthol. The study reinforces the need to select representative sentinel species from different habitats for biomonitoring purposes and provides further support for the use of biomarkers in assessing the health of coastal areas.
Environmental pollution (Barking, Essex: 1987) 01/2008; 155(2):327-35. · 3.43 Impact Factor
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ABSTRACT: Cultured and wild sea bass (Dicentrarchus labrax) from the Arade Estuary were sampled in summer and winter and the degree of exposure to metals and polycyclic aromatic hydrocarbons (PAHs) assessed, together with some biochemical responses against those and other pollutants. The highest levels of copper (up to 997 microg g-1 dry weight) and cadmium (up to 4.22 microg g-1 dry weight) were detected in the liver and kidney of cultured specimens, whereas the highest exposure to PAHs was observed in wild fish. Significant alterations in some biochemical markers were detected and associated to pollutant exposure. Thus, metallothionein concentrations were higher in the tissues of cultured fish and positively correlated with metal residues. The activity 7-ethoxyresorufin O-deethylase ranged from 28 pmol/min/mg protein in cultured fish to 83 pmol/min/mg protein in wild fish collected near a marina area. Cultured fish and wild fish from the marina area had depressed acetylcholinesterase in muscle tissue and a parasitic infection in the gonads. The obtained results support the usefulness of the combined use of chemical and biochemical markers to assess the impact of anthropogenic pollutants in both wild and cultured fish.
Environmental Research 03/2007; 103(2):247-56. · 3.40 Impact Factor
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ABSTRACT: Carps (Cyprinus carpio) and red swamp crayfish (Procambarus clarkii) were sampled from two lowstream Mediterranean rivers (Anoia and Cardener) receiving extensive urban and industrial waste water discharges. Tissue residues of selected pollutants (organochlorinated compounds, polycyclic aromatic hydrocarbons (PAHs)) were determined in conjunction with different biochemical responses (cytochrome P450, phase II enzymes) with the aim of investigating whether resident organisms were responsive to changes in water quality. Biota inhabiting those rivers were highly exposed to complex mixtures of polychlorobiphenyls and dichlorodiphenyltrichloroethanes (up to 19 ng/g w.w.) and PAHs (up to 6097 ng/g of hydroxylated PAHs in bile), the highest residues being observed in carps from Cardener. This has a reflection on 7-ethoxyresorufin O-deethylase (EROD) activity; that in carps from Cardener ranged between 350 and 550 pmol/min/mg protein, whereas in carps from Anoia ranged between was 90 and 250 pmol/min/mg protein. The highest activity recorded was downstream of the sewage treatment plants in both rivers. Phase II enzymes were less sensitive to polutant exposure than EROD; nonetheless, both glutathione S-transferase and UDP-glucuronyl transferase were higher in carps from Cardener. The results support the usefulness of the combined use of chemical and biochemical responses to assess and diagnose pollution in highly stressed ecosystems.
Environmental Research 11/2002; 90(2):169-78. · 3.40 Impact Factor
