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.

Phthalates and perfluorinated alkylated substances in Atlantic bluefin tuna (Thunnus thynnus) specimens from Mediterranean Sea (Sardinia, Italy): Levels and risks for human consumption.

Atlantic blue fin tuna (Thunnus thynnus) is a species of great importance for Mediterranean Sea area, from both ecological and commercial points of view. The scientific literature reports few data on the contamination of this fish by emerging organic compounds such as perfluorinated alkylated substances(PFASs) and phthalates, being the latter never been studied in tuna. This study therefore investigated the presence of the PFASs perfluorooctane sulphonate (PFOS) and perfluoroctanoic acid (PFOA) and the phthalate di-2-ethylhexyl phthalate (DEHP), also monitored by its metabolite mono-2-ethylhexyl phthalate(MEHP), to assess both the state of contamination of Atlantic bluefin tuna specimen and the risk due to the toxicity of these compounds for human consumption. While PFOA was never found, detectable levels of PFOS (0.4-1.88 ng/g), DEHP (9-14.62 ng/g) and MEHP (1.5-6.30 ng/g) were found. The results were elaborated relating the accumulation to the size and age of the individuals and showed a correlation between the levels of different pollutants investigated.

Pilot study on levels of chemical contaminants and porphyrins in Caretta caretta from the Mediterranean Sea.

Perfluorinated compounds (PFCs), synthetic musks compounds (SMCs), bisphenol A (BPA), para-nonylphenol (p-NP) and polybrominated diphenyl ethers (PBDEs) are known for their toxicity and ability to interfere with the endocrine system. The aim of this study was to determine levels and distribution of the above mentioned compounds in liver samples of Caretta caretta and levels of porphyrins that have been proposed as sensitive biomarkers of exposure to contaminants. This paper reports the results for 9 specimens yet analysed. Musk ketone was never detected, PFOA was found in one sample, while PFOS was the prevalent contaminant. For PFCs the levels are lower than the results of studies of comparison. The porphyrins profile showed a predominance of protoporphyrins on coproporphyrins and uroporphyrins, with a positive statistical correlation between levels of PFOS and uroporphyrins. These data represent, for several parameters, the first evidence of contaminant levels and biomarker responses in loggerhead turtles.

Differential ABCB and ABCC gene expression and efflux activities in gills and hemocytes of Mytilus galloprovincialis and their involvement in cadmium response.

The aim of the study was to characterize ABC transport proteins gene expression and efflux activities in gills and hemocytes of the Mediterranean mussel and to evaluate their response to Cd. At basal level a higher expression of abcb-like gene was observed in gills than in hemocytes while abcc-like gene showed similar levels. Both P-gp and MRPs inhibitors (cyclosporine and MK571) blocked efflux activities in gills; hemocytes were sensitive only to MK571. After 120 min in vitro pre-exposure to CdCl2, the efflux activity increased significantly in gills and hemocytes. In vivo exposure to CdCl2 (0.4 µM) increased abcb-like gene expression in gills without affecting efflux activity. In hemocytes abcc-like gene resulted up-regulated and Ca-AM efflux resulted enhanced. An increased uptake of Cd in gills biopsies was observed in the presence of both P-gp and MRPs inhibitors. Our results indicate that ABC transporters seem involved in the first protective response to Cd and this response is tissue-specific.

Levels and spatial distribution of perfluorinated compounds (PFCs) in superficial sediments from the marine reserves of the Tuscan Archipelago National Park (Italy).

Surface sediments from marine reserves of the Tuscan Archipelago National Park (Italy), including remote islands not directly affected by anthropogenic influences, were analyzed for two dominant perfluorinated compounds (PFCs), namely perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA), using LC-ESI-MS/MS. The concentrations of PFOS were low, with medians ranging from 0.25 to 1.50 ng/g dry wt, whereas PFOA was always below the detection limit of 0.1 ng/g dry wt. The present paper can be considered a baseline study useful in future environmental monitoring programs.

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.

Perfluorinated compounds in blood of Caretta caretta from the Mediterranean Sea.

Perfluorinated compounds (PFCs), widely used for their hydro-oil repellent properties, are almost non-degradable in the environment; there is scientific evidence that indicate bioaccumulation. They represent a threat to many organisms, because they are toxic and are endocrine disruptors. Scientific studies have demonstrated the presence of PFCs in blood and liver samples of fish, turtles, birds and mammals of marine ecosystems in different geographical areas. The aim of this study was to determine the distribution of PFOS and PFOA in blood samples of the marine turtle Caretta caretta, using a minimally invasive sampling procedure. 49 blood samples of marine turtle, taken from several Italian marine turtle rescue centers, were analyzed. While PFOA was never detected, measurable concentrations of PFOS were found in 15 blood samples; the values show a range from 1.14 ng/g to 28.51 ng/g (wet wt.). No differences between groups of samples taken from different areas were found.

The influence of endocrine disruptors in a selected population of infertile women.

Several studies report that endocrine disrupting chemicals (EDC) able to interfere with endocrine homeostasis may affect women's reproductive health. We analyzed EDC serum levels and nuclear receptors (NRs) expression in order to have an indication of the internal dose of biologically active compounds and a measurement of indicators of their effects, as a result of the repeated uptake from environmental source. The percentage of patients with detectable bisphenol A (BPA) concentrations was significantly higher in the infertile patients compared with fertile subjects. No significant difference was found between the groups with regard to perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), mono-ethylhexyl phthalate (MEHP) and di-(2-ethylhexyl) phthalate (DEHP) concentrations. Among infertile women, the mean expression of estrogen receptor alpha (ERα) and beta (Erß), androgen receptor (AR) and pregnane X receptor (PXR) was significantly higher than fertile patients. The mean expression of aryl hydrocarbon receptor (AhR) and peroxisome proliferator-activated receptor gamma (PPARγ) did not show significant differences between two groups. Patients with endometriosis had higher levels of PPARγ than all women with other causes of infertility. This study led further support to EDC exposure as a risk factor for women's fertility.

Alteration of potential harmful elements levels in sediments and biota from the central Mediterranean Sea (Aeolian Archipelago) following an episode of intense volcanic activity.

In this paper levels of four (Hg, Cd, Pb, As) potential harmful elements (PHE) were measured in three different environmental matrices (sediments, macroalgae and fishes) from the Aeolian Archipelago and control areas both after 1 and 10 months from a volcanic activity of particular relevance occurred at the end of October 2002. Results were analysed on a multivariate statistical basis with the aim to evaluate: (I) general levels of pollution and increase of PHE due to the event; (II) differences observed among tested matrices in the time of recovery after the occurrence of the critical event; (III) the biological enrichment of PHE along the trophic web produced by the geological event. Results evidenced that volcanic emissions could represent a local source of particular relevance able to determine great enrichments of considered PHE in sediments and biological species. After 10 months from the event, levels in sediments and macroalgae notably decreased, whereas fish species evidenced an increase, principally related to the bioaccumulation phenomena. On the basis of the biological enrichment factors (BEF), major enrichments were evidenced after 1 month whereas, after 10 months, were recorded values reliable to an incomplete recovery. Concerning Cd, the BEF higher levels reported for the species Serranus cabrilla was probably related both to the diet and to the specific detoxification rates of this species.

Acute toxicity and biodegradability of N-alkyl-N-methylmorpholinium and N-alkyl-DABCO based ionic liquids.

N-alkyl-N-methylmorpholinium and N-alkyl substituted 1,4-diazabicyclo[2.2.2]octane (DABCO) based ionic liquids (ILs), N-alkyl-DABCO, bearing short alkyl chains are characterised by a low toxicity to Vibrio fischeri, although toxicity significantly increases on increasing the alkyl chain length. Alkyl chain length affects also biodegradability in the 28 days tests; the higher level of biodegradation was found in both the series in the case of the ethyl (C2) derivatives. In the case of N-ethyl DABCO based IL, although biodegradability is still around 40%, and consequently this IL cannot be classified as "readily biodegradable", this value is similar to the more biodegradable functionalized imidazolium based ILs.

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.

Differences in Pop Levels between Conventional and Omega-3 Fatty Acid-Enriched Milk and Dairy Products.

Conventional and omega-3 fatty acid-enriched milk and cheese were analyzed for persistent organic pollutants (POPs). Omega-3-enriched products are usually supplemented with fish oil which is potentially contaminated. All classes of the considered POPs (PCBs, DDT, HCB, PBDEs, and PCDD/Fs) were found in the samples, with average concentrations higher in omega-3-enriched products than in conventional ones. For PCBs, DDT, and HCB, differences were statistically significant and, therefore, cannot be ascribed to normal variability. With regard to all classes of compounds, the highest levels in individual samples were always found in omega-3 products, in line with the hypothesis that these foods are potentially more contaminated than conventional ones.