Polymer composition analysis of plastic debris ingested by loggerhead turtles (Caretta caretta) in Southern Tyrrhenian Sea through ATR-FTIR spectroscopy.

The ingestion of anthropogenic plastic debris by marine wildlife is widespread in the Mediterranean Sea. The endangered status (in the IUCN Red List) of Loggerhead turtle (Caretta caretta, Linnaeus, 1758) is a consequence of its vulnerability. In this study, macro-/meso-plastics (5-170 mm) collected from faeces of twelve loggerhead turtles rescued (live) in the Aeolian Archipelago (Southern Tyrrhenian Sea, Italy) were analyzed by size, weight, shape, color and polymer type through Attenuated Total Reflectance Fourier-Transform Infrared Spectroscopy (ATR-FTIR). The defecation rate during hospitalization (7-14 days) varied among turtles (from 0.08 to 0.58). The mean number of plastic expulsions (2.7 ± 1.8 items for turtle) was higher during the 5th day of hospitalization (Kruskal-Wallis test, P = 0.01). However, the mean number of plastic-like items defecated during the common days of hospitalization did not vary among turtles (Kruskal-Wallis test, P > 0.05). All turtles were found to have ingested plastic. A total of 114 debris items were recovered from their faeces, 113 of which were identified as plastic. Their color was mostly white-transparent (64.9%) and light (19.3%). Shape was mainly fragments (52.6%), sheets (38.6%), followed by nylon, net-fragments, elastic plastic, foamed plastic and industrial granules (8.8%). Meso-plastics (5-25 mm) represented 72% of the total number of debris and were found more frequently in turtle with Curved Carapace Length (CCL) ≤ 60 cm (CCL = 30-60 cm, n = 5) than those with CCL >60 cm (CCL = 60-71 cm, n = 7). Plastic items were composed mainly of polyethylene (48.2%) and polypropylene (34.2%). Polypropylene (R2 = 0.95, P < 0.001) and polyisoprene (R2 = 0.45, P = 0.017) were more common in meso-plastics while polyethylene (R2 = 0.44, P < 0.01) in macro-plastics. Finally, high-density polyethylene, polyvinyl chloride, polyamide and polyurethane were also found in some turtles. This study reveals high spreads of plastic contamination in faeces of both turtles with CCL ≤60 cm and CCL >60 cm, particularly vulnerable to the increasing quantity of floating plastic into their foraging sites highlighting the need of further research to associate debris ingestion with turtle diet and their size.

Phthalate esters (PAEs) concentration pattern reflects dietary habitats (δ13C) in blood of Mediterranean loggerhead turtles (Caretta caretta).

Phthalic acid esters (PAEs) are classified as endocrine disruptors, but it remains unclear if they can enter the marine food-web and result in severe health effects for organisms. Loggerhead turtles (Caretta caretta) can be chronically exposed to PAEs by ingesting plastic debris, but no information is available about PAEs levels in blood, and how these concentrations are related to diet during different life stages. This paper investigated, for the first time, six PAEs in blood of 18 wild-caught Mediterranean loggerhead turtles throughout solid-phase extraction coupled with gas chromatography-ion trap/mass spectrometry. Stable isotope analyses of carbon and nitrogen were also performed to assess the resource use pattern of loggerhead turtles. DEHP (12-63 ng mL-1) and DBP (6-57 ng mL-1) were the most frequently represented PAEs, followed by DiBP, DMP, DEP and DOP. The total PAEs concentration was highest in three turtles (124-260 ng mL-1) whereas three other turtles had concentrations below the detection limit. PAEs were clustered in three groups according to concentration in all samples: DEHP in the first group, DBP, DEP, and DiBP in the second group, and DOP and DMP in the third group. The total phthalates concentration did not differ between large-sized (96.3 ± 86.0 ng mL-1) and small-sized (67.1 ± 34.2 ng mL-1) turtles (p < 0.001). However, DMP and DEP were found only in large-sized turtles and DiBP and DBP had higher concentrations in large-sized turtles. On the other hand, DEHP and DOP were found in both small- and large-sized turtles with similar concentrations, i.e. ~ 21.0/32.0 ng mL-1 and ~ 7.1/9.9 ng mL-1, respectively. Winsored robust models indicated that δ13C is a good predictor for DBP and DiBP concentrations (significant Akaike Information criterion weight, AICwt). Our results indicate that blood is a good matrix to evaluate acute exposure to PAEs in marine turtles. Moreover, this approach is here suggested as a useful tool to explain the internal dose of PAEs in term of dietary habits (δ13C), suggesting that all marine species at high trophic levels may be particularly exposed to PAEs, despite their different dietary habitats and levels of exposure.

Fast and Reliable Determination of Phthalic Acid Esters in the Blood of Marine Turtles by Means of Solid Phase Extraction Coupled with Gas Chromatography-Ion Trap/Mass Spectrometry.

The presence of phthalic acid esters (PAEs) in marine environments is an important issue. These chemicals are able to affect marine organisms, particularly marine turtles, and to act as endocrine disrupters. In this paper, for the first time, a simple and reproducible analytical method based on solid-phase extraction (SPE) coupled with gas chromatography-ion trap/mass spectrometry (GC-IT/MS) was developed for the extraction of phthalates from the blood of marine turtles. The extraction was obtained by using C18 phthalates-free as the stationary phase. In order to individuate the best working conditions for the extraction, the adsorption isotherms and breakthrough curves were studied. The overall analytical methodology was validated in terms of limit of detection (LOD, 0.08-0.6 ng mL-1), limit of quantification (LOQ, 0.4-0.8 ng mL-1), and correlation coefficients (>0.9933). By using this procedure, percentage recoveries ranging from 89 to 103% were achieved. The precision parameters (intra-day and inter-day) were studied, and the obtained values were smaller than 12.5%. These data confirm the goodness of the proposed analytical methodology, which is applied to real samples.

Antibiotic Resistance of Gram-Negative Bacteria from Wild Captured Loggerhead Sea Turtles.

Sea turtles have been proposed as health indicators of marine habitats and carriers of antibiotic-resistant bacterial strains, for their longevity and migratory lifestyle. Up to now, a few studies evaluated the antibacterial resistant flora of Mediterranean loggerhead sea turtles (Caretta caretta) and most of them were carried out on stranded or recovered animals. In this study, the isolation and the antibiotic resistance profile of 90 Gram negative bacteria from cloacal swabs of 33 Mediterranean wild captured loggerhead sea turtles are described. Among sea turtles found in their foraging sites, 23 were in good health and 10 needed recovery for different health problems (hereafter named weak). Isolated cloacal bacteria belonged mainly to Enterobacteriaceae (59%), Shewanellaceae (31%) and Vibrionaceae families (5%). Although slight differences in the bacterial composition, healthy and weak sea turtles shared antibiotic-resistant strains. In total, 74 strains were endowed with one or multi resistance (up to five different drugs) phenotypes, mainly towards ampicillin (~70%) or sulfamethoxazole/trimethoprim (more than 30%). Hence, our results confirmed the presence of antibiotic-resistant strains also in healthy marine animals and the role of the loggerhead sea turtles in spreading antibiotic-resistant bacteria.

Sub-Chronic Oral Exposure to Iridium (III) Chloride Hydrate in Female Wistar Rats: Distribution and Excretion of the Metal.

Iridium tissue distribution and excretion in female Wistar rats following oral exposure to iridium (III) chloride hydrate in drinking water (from 1 to 1000 ng/ml) in a sub-chronic oral study were determined. Samples of urine, feces, blood and organs (kidneys, liver, lung, spleen and brain) were collected at the end of exposure. The most prominent fractions of iridium were retained in kidney and spleen; smaller amounts were found in lungs, liver and brain. Iridium brain levels were lower than those observed in other tissues but this finding can support the hypothesis of iridium capability to cross the blood brain barrier. The iridium kidney levels rose significantly with the administered dose. At the highest dose, important amounts of the metal were found in serum, urine and feces. Iridium was predominantly excreted via feces with a significant linear correlation with the ingested dose, which is likely due to low intestinal absorption of the metal. However, at the higher doses iridium was also eliminated through urine. These findings may be useful to help in the understanding of the adverse health effects, particularly on the immune system, of iridium dispersed in the environment as well as in identifying appropriate biological indices of iridium exposure.

Human biomonitoring for metals in Italian urban adolescents: data from Latium Region.

As a part of the activities of the first Italian human biomonitoring survey (PROBE - PROgramme for Biomonitoring general population Exposure), a reference population of adolescents, aged 13-15 years, was examined for their exposure to metals. The study included 252 adolescents living in urban areas, representative of Latium Region (Italy) and blood specimens were analyzed for metals (As, Be, Cd, Co, Cr, Hg, Ir, Mn, Mo, Ni, Pb, Pd, Pt, Rh, Sb, Sn, Tl, U, V and W) by sector field inductively coupled plasma mass spectrometry. The results obtained will improve the knowledge about the body burden in adolescents and are tentative reference values for Italian young people as a basis for risk evaluation deriving from urban/environmental exposure to metals.

Uncertainty evaluation in the analysis of biological samples by sector field inductively coupled plasma mass spectrometry. Part B: measurements of As, Co, Cr, Mn, Mo, Ni, Sn and V in human serum.

This protocol was developed to estimate the uncertainty of measurements in the sector field inductively coupled plasma mass spectrometry analysis of As, Co, Cr, Mn, Mo, Ni, Sn and V in human serum by utilizing in-house validation data. The approach is outlined in the Eurachem/Citac Guide and other relevant publications. The results generated were the followings: (i) coefficients of regression >0.9995 over two orders of magnitude of metal concentration; (ii) method quantification limits between 0.05 µg/L (Cr, Mn) and 0.49 µg/L (As); (iii) mean trueness checked against a certified reference material between 95.4% (As) and 107.7% (Ni); (iv) repeatability better than 10.2% over the range 0.1-2.0 µg/L; (v) reproducibility better than 12.0% over the range 0.1-2.0 µg/L; and (vi) expanded uncertainty budget comprised between 14.7% (Mn) and 27.9% (Cr) over the range 0.1-2.0 µg/L.

Italian network for human biomonitoring of metals: preliminary results from two Regions.

The Italian program for human biomonitoring (HBM) of chemical elements, PROgram for Biomonitoring of the Exposure (PROBE), started in 2008 with the aim to provide the knowledge about risk assessment of the Italian population following the environmental exposure to metals. The project is implemented through a HBM campaign for the production of data on 19 metals in the blood and serum of subjects living in different Italian Regions. The metals studied are: antimony, beryllium, cadmium, chromium, cobalt, iridium, lead, manganese, mercury, molybdenum, nickel, palladium, platinum, rhodium, thallium, tin, tungsten, uranium and vanadium. The first phase of the project has included the development and validation of laboratory protocols for the collection of fluids and quantification of metals. The second phase provides the HBM data expressed as the reference values (RVs) for the Italian population, i.e., as the level of metals in the general population not occupationally exposed. In this paper, the experimental protocols used for the maintenance of high standards of analysis and the RVs for metals in serum of inhabitants of two Italian Regions (Calabria and Umbria) are described.

Uncertainty evaluation in the analysis of biological samples by sector field inductively coupled plasma mass spectrometry. Part A: Measurements of Be, Cd, Hg, Ir, Pb, Pd, Pt, Rh, Sb, U, Tl and W in human serum.

A protocol that utilises data (trueness/recovery, precision and robustness) from validation tests to calculate measurement uncertainty was described and applied to a sector field inductively coupled plasma mass spectrometry (SF-ICP-MS)-based method for the determination of Be, Cd, Hg, Ir, Pb, Pd, Pt, Rh, Sb, U, Tl and W in human serum. The method was validated according to criteria issued by international bodies such as AOAC, Eurachem and ISO and the uncertainty in the analytical measurements was estimated following the Eurachem/Citac guide. The methodology was based on dilution of human serum with water and analysis by serum-matched standard calibration. The method quantification limits ranged 0.02 microg/L (Tl, Ir) to 0.26 microg/L (Hg). The coefficients of regression were greater than 0.9991 over a range of two orders of magnitude of concentration. The mean trueness was 101% and the mean recovery on three levels of fortification (1-, 1.5-, and 2-times the baseline serum level) ranged between 93.3% and 106%. The maximum relative standard deviation values for repeatability and within-laboratory reproducibility were 12.8% and 13.5%. The method was robust to slight variations of some critical factors relevant to the sample preparation and SF-ICP-MS instrumentation. The relative expanded uncertainty over three levels of concentration ranged from 11.6% (Hg) to 27.6% (Pt), and the uncertainty on the within-laboratory reproducibility, which included factors such as time, analyst and calibration, represented the main contribution to the overall uncertainty.

Health risk evaluation associated to Planktothrix rubescens: An integrated approach to design tailored monitoring programs for human exposure to cyanotoxins.

Increasing concern for human health related to cyanotoxin exposure imposes the identification of pattern and level of exposure; however, current monitoring programs, based on cyanobacteria cell counts, could be inadequate. An integrated approach has been applied to a small lake in Italy, affected by Planktothrix rubescens blooms, to provide a scientific basis for appropriate monitoring program design. The cyanobacterium dynamic, the lake physicochemical and trophic status, expressed as nutrients concentration and recycling rates due to bacterial activity, the identification/quantification of toxic genotype and cyanotoxin concentration have been studied. Our results indicate that low levels of nutrients are not a marker for low risk of P. rubescens proliferation and confirm that cyanobacterial density solely is not a reliable parameter to assess human exposure. The ratio between toxic/non-toxic cells, and toxin concentrations, which can be better explained by toxic population dynamic, are much more diagnostic, although varying with time and environmental conditions. The toxic fraction within P. rubescens population is generally high (30-100%) and increases with water depth. The ratio toxic/non-toxic cells is lowest during the bloom, suggesting a competitive advantage for non-toxic cells. Therefore, when P. rubescens is the dominant species, it is important to analyze samples below the thermocline, and quantitatively estimate toxic genotype abundance. In addition, the identification of cyanotoxin content and congeners profile, with different toxic potential, are crucial for risk assessment.

A study on metals content in patients with colorectal polyps.

Data on metals involvement in colorectal polyps are scarce and fragmentary. The aim of this study was to examine whether the level of metals could be associated with risk of colorectal polyp development. The concentration of 15 chemical elements (Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg Mg, Mn, Pb, Se, Sr, and Zn) in 17 colorectal biopsies of healthy individuals, in 15 polypotic and corresponding nonpolypotic biopsies taken from the same individual, was evaluated. Concentration in polyps of metals such as Al, Ca, Mg, Mn, Pb, Sr, and Zn was unchanged both in unpaired and paired samples; elements such as Ba, Cd, and Hg were significantly lower and Fe was significantly higher both in individual and paired tissues. Cobalt, Cr, and Cu were significantly different only between polyps and the adjacent normal tissue area; Se showed a significant accumulation comparing polyps versus healthy tissues. The difference found in some elements between polyps and a control tissue provides an indication about the role of essential and nonessential elements in the early stage (polyps) in the colon carcinogenic process and encourages further studies to confirm the involvement of such elements in neoplastic processes.

Determination of 30 elements in colorectal biopsies by sector field inductively coupled plasma mass spectrometry: method development and preliminary baseline levels.

An analytical procedure applicable to restricted sample sizes was developed and applied to the analysis of 30 chemical elements in colorectal biopsies of healthy patients. Acidic microwave digestion processed