Biomonitoring of chemicals in biota of two wetland protected areas exposed to different levels of environmental impact: results of the "PREVIENI" project.

The PREVIENI project (funded by the Ministry of Environment) investigated the exposure to endocrine disrupters in samples of human population and environmental biota in Italy. The environmental biomonitoring considered two Italian WWF Oasis, with the aim to compare the presence and effects of endocrine disruptors in organisms from two protected natural areas, respectively, upstream and downstream a chemical emission site. Chemical analysis of pollutants' tissue levels was made on tissues from earthworm, barbell, trout, and coot, selected as bioindicator organisms. The contaminants considered were as follows: the perfluorinated compounds perfuoroctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), polychlorinated biphenyls (PCBs 58 congeners), polybrominated diphenyl ethers (PBDEs, 13 congeners), polycyclic aromatic hydrocarbons (PAHs, 16 compounds), toxic trace elements, the phthalate di-2-ethylexyl phthalate (DEHP) and its primary metabolite, bisphenol A, synthetic musk compounds (musk xylene, musk ketone, tonalide, and galaxolide), and p-nonylphenol. The analyses showed low concentrations of most pollutants in all species from both areas, compared to available literature; noticeable exceptions were the increases of DEHP's primary metabolite, PBDE, PAHs, Hg, and Pb in barbells, and of PCB and Cd in earthworms from the downstream area. The results showed the presence of endocrine disruptors, including those considered as "non-persistent," in bioindicators from protected areas, albeit at low levels. The results provide a contribution to the evaluation of reference values in biota from Mediterranean Europe and support the relevance of monitoring exposure to pollutants, in particular for freshwater environment, also in protected areas.

Occurrence of PCDD/PCDFs and PCBs in soil and comparison with CYP1A response in PLHC-1 cell line.

The responsiveness of CYP1A (gene transcription and EROD enzyme activity) in the cell line Poeciliopsis lucida hepatoma (PLHC-1) upon exposure to extracts of contaminated soil samples was investigated and compared to levels of PCDD/PCDFs and PCBs including non-ortho obtained by GC/MS analysis. Soil samples A and B were collected in sites A and B. Two fractions, not purified (np) and purified (p), were obtained from each sample and analyzed for PCDD/PCDF and PCB content by GC/MS; in parallel they were tested for 24 h with PLHC-1. CYP1A response was investigated at gene (RT-qPCR) level and as 7-ethoxyresorufin-O-deethylase (EROD) enzyme activity. Chem-TEQs and Bio-TEQs were then calculated. ∑TEQ calculated for PCDD/Fs and PCBs was 0.081 pg/g and 20.32 pg/g for samples A and B, respectively. PLHC-1 showed less up-regulation of cyp1a gene on exposure to the two purified fractions (Ap 2.1-fold and Bp 1.8-fold) than to non-purified fractions (up to 15-fold for Anp and 13-fold for Bnp). EROD was also induced 2.38- and 9.44-fold in the two purified fractions (Ap and Bp) compared to model inducer 2,3,7,8-TCDD, and up to 16.03-fold for non-purified Anp and 33.79-fold for Bnp. The combination of CYP1A response, obtained in a PLHC-1 cell-based bioassay, with contaminant residue analysis provided a better description of the presence and toxicity of dioxin-like compounds in an environmental matrix.

Records of polychlorinated biphenyls (PCBs) in sediments of four remote Chilean Andean Lakes.

Sediment cores from four Chilean lakes along the Andes Chain (Chungará, Laja, Castor and Venus) were analysed in order to investigate PCB concentrations and distributions in sediment samples. Sediment cores were analysed for PCBs using gas chromatography (GC-ECD/MS) and radioisotopically dated using 210Pb. Organic carbon content (OC) and 210Pb fluxes were also measured. Results showed that sediment PCB concentrations (ngg(-1) d.w.) at Lake Chungará (1.2 +/- 1) in northern Chile, Laja (5 +/- 4) in central, and in Lake Castor (3.5 +/- 4) in southern Chile (the eastern side of the Andes Mountain) were lower than sediments collected from Lake Venus (64 +/- 30) located in southern Chile (the western side) which contained 15-fold higher concentrations of PCBs. The percentage (%) of organic carbon was variable and showed a high range of values in the sediment fluctuating from 2% (Lake Laja) to 22% (Lake Chungará). Analysis of 210Pb fluxes, presented a decrease trend following Lake Laja>Castor>Chungará with a positive correlation with rainfall at each site. Sedimentation rates in Lake Castor (1846 gm(-2)yr(-1)) were higher than at Chungará (748 gm(-2)yr(-1)) and Lake Laja (508 gm(-2)yr(-1)). Focusing factor (FF) is used as a tool to elucidate PCB input in the aquatic ecosystem. FF were lower (<1) for the shallower lakes (Lakes Chungará and Castor). This study provides background levels of PCBs at remote lakes in Chile. Differences in geographical characteristics (orographic effect) might play an important role in the arrival of PCBs, particularly into the southern lakes. PCB fluxes indicated deposition of PCBs in recent sediments is higher than in previous years with peaks of PCB between 1991 and 1998. The continuing increase of PCB inputs in remote Chilean lakes, could be associated with long range atmospheric transport (LRAT).

Influence of titanium dioxide nanoparticles on 2,3,7,8-tetrachlorodibenzo-p-dioxin bioconcentration and toxicity in the marine fish European sea bass (Dicentrarchus labrax).

The present study investigated the influence of nano-TiO(2) (1 mg L(-1)) on 2,3,7,8-tetrachlorodibenzo-p-dioxin(2,3,7,8-TCDD) (46 pg L(-1)) bioconcentration and toxicity in the European sea bass (Dicentrarchus labrax) during 7 days in vivo exposure. A multimarkers approach was applied in different organs: detoxification in liver; innate immunity and pro-inflammatory response and adaptive immunity in gills and spleen; genotoxicity in peripheral erythrocytes and muscle. Bioconcentration of 2,3,7,8-TCDD in presence of nano-TiO2 was investigated in liver, skin and muscle as well as interaction between nano-TiO2 and organic pollutants in artificial sea water (ASW). Nano-TiO2 negatively influenced immune response induced by 2,3,7,8-TCDD in spleen but not in gills and reduced the DNA damage induced by 2,3,7,8-TCDD in erythrocytes. nano-TiO2 did not interfere with 2,3,7,8-TCDD detoxification and bioconcentration according to the observed no interaction of the nano-TiO2 with organic pollutants in ASW.

Induction of CYP1A and ABC transporters in European sea bass (Dicentrarchus labrax) upon 2,3,7,8-TCDD waterborne exposure.

The aim of this study was to characterize the responsiveness of CYP1A and ABC transport proteins in European Sea bass (Dicentrarchus labrax) waterborne exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) (46 pg/L) for 24 h and 7 days. Genes modulation (abcb1, abcc1-2, cyp1a), EROD activity were investigated in liver and 2,3,7,8-TCDD bioconcentration in liver and muscle. TCDD induced significantly cyp1a gene expression and EROD activity at 24 h and 7 d. A significant up-regulation of abcb1 was also observed but only after 7 days. No modulation of abcc1 and abcc2 genes was observed. Waterborne TCDD exposure was able to induce CYP1A and abcb1 encoding for P-glycoprotein in juvenile of European sea bass.

Levels of Persistent Organic Pollutants (POPs) in sediments from Lenga estuary, central Chile.

Superficial sediments from Lenga estuary in the VIII region of central Chile were analysed for Polychlorinated Biphenyls (PCBs) and Hexachlorobenzene (HCB). Organic carbon (OC) analysis was also performed and the statistical correlations (Pearson correlations) between compounds were also tested. Results showed the total PCB concentrations (ng g(-)(1)d.w.) fluctuated from ∼ 20 to 10,000. These levels are higher than those reported in other coastal sediments of the world. The PCB profile was dominated by middle to highly chlorinated congeners (6-Cl=40% and 7-Cl=30%). HCB concentrations (ng g(-)(1)d.w.), were lower than the PCBs and, fluctuated between 1 (at L1) and 870 (at L4). OC values ranged from 1% (L1) to 7% (L4) showing a significant positive correlations between OC% and PCBs (r=0.86; p<0.05) and for HCB (r=0.71). These results are an important contribution to knowledge of levels of POPs levels in coastal Chilean environments.

Interactive effects of n-TiO2 and 2,3,7,8-TCDD on the marine bivalve Mytilus galloprovincialis.

Despite the growing concern over the potential biological impact of nanoparticles (NPs) in the aquatic environment, little is known about their interactions with other pollutants. The bivalve Mytilus sp, largely utilized as a sentinel for marine contamination, has been shown to represent a significant target for different types of NP, including n-TiO2, one of the most widespread in use. In this work, the possible interactive effects of n-TiO2 and 2,3,7,8-TCDD, chosen as models of NP and organic contaminant, respectively, were investigated in Mytilus galloprovincialis. In vitro experiments with n-TiO2 and TCDD, alone and in combination, were carried out in different conditions (concentrations and times of exposure), depending on the target (hemocytes, gill cells and biopsies) and the endpoint measured. Mussels were also exposed in vivo to n-TiO2 (100 µg L(-1)) or to TCDD (0.25 µg L(-1)), alone and in combination, for 96 h. A wide range of biomarkers, from molecular to tissue level, were measured: lysosomal membrane stability and phagocytosis in hemocytes, ATP-binding cassette efflux transporters in gills (gene transcription and efflux activity), several biomarkers of genotoxicity in gill and digestive cells (DNA damage, random amplified polymorphic DNA-RAPD changes), lysosomal biomarkers and transcription of selected genes in the digestive gland. The results demonstrate that n-TiO2 and TCDD can exert synergistic or antagonistic effects, depending on experimental condition, cell/tissue and type of measured response. Some of these interactions may result from a significant increase in TCDD accumulation in whole mussel organisms in the presence of n-TiO2, indicating a Trojan horse effect. The results represent the most extensive data obtained so far on the sub-lethal effects of NPs and organic contaminants in aquatic organisms. Moreover, these data extend the knowledge on the molecular and cellular targets of NPs in bivalves.

Modulation of CYP1A and genotoxic effects in European seabass (Dicentrarchus labrax) exposed to weathered oil: a mesocosm study.

The aim of this study was to assess medium-term toxicity of weathered oil on European seabass. A mesocosm system reproducing an oil spill at sea was applied. Fish were collected after 48 h, 7, 30 and 60 days. Cyp1a gene transcription, EROD and UDPGT activities, bile PAHs metabolites and micronuclei frequency were investigated. A progressive disappearance of low molecular weight n-alkanes and PAHs in the water of the mesocosm occurred during the experimentation. Fishes exposed to oil displayed a significant increase of cyp1a expression and EROD activity during the entire experiment as well as higher concentrations of PAHs metabolites in bile. Micronulei frequency resulted significantly higher during all experiment in oil exposed sea bass compared to controls. The results highlight the environmental risk associated with the release of oil products at sea and confirm the adopted parameters as useful tools for studying the impact of accidental oil spills on fish.

Persistent organic pollutants in human milk from central Italy: levels and time trends.

Persistent organic pollutants (POPs) such as HCB, p,p'-DDE, and PCBs were measured in Italian breast milk. This work is part of a study on human milk, adipose tissues, and food carried out in the same area over the last 20 years. The results showed the prevalence of p,p'-DDE and PCBs over HCB. Comparison of our results with those of previous studies carried out in the same area showed that concentrations are decreasing. No statistically significant differences in organochlorine levels were found when the samples were divided into maternal age classes and into the categories "primiparae" and "multiparae". In order to quantify the amount of the molecules of interest transmitted by mother to child during breast feeding, we estimated the daily intake of each class of compounds: our results indicated that HCB and p,p'-DDE were several times lower than the safety thresholds.

Biomonitoring of polybrominated diphenyl ether (PBDE) pollution: a field study.

Polybrominated diphenyl ethers (PBDEs) and cytochrome P450 enzyme activities were investigated in European eels (Anguilla anguilla) collected from seven sites in a coastal lagoon in the north-western Mediterranean Sea, Orbetello lagoon (Italy). Twelve PBDE congeners were measured in muscle and two CYP1A enzyme activities, 7-ethoxyresorufin-O-deethylase (EROD) and benzo(a)pyrene monooxygenase (BP(a)PMO), were investigated in liver microsomal fraction in order to obtain insights into the health of the lagoon environment. PBDE muscle levels were low and the most abundant congeners were 2,2',4,4'-tetrabromodiphenylether (BDE-47), 2,2',4,4',5,5'-hexaBDE (BDE-153) and 2,2',4,5'-tetraBDE (BDE-49). EROD and B(a)PMO activities were also low and no differences were observed between eels from different sites. Multivariate analysis (PCA) did not indicate correlations between PBDEs and either P450 activities.