Imposex in Nassarius nitidus (Jeffreys, 1867) as a possible investigative tool to monitor butyltin contamination according to the Water Framework Directive: A case study in the Venice Lagoon (Italy).

Butyltin (TBT, DBT, and MBT) effects on molluscs, especially endocrine disruption in bivalves and gastropods, have been widely investigated. Imposex, the superimposition of male characters onto female gonochoristic Caenogastropods, is the most studied biological effect of TBT. TBT compounds are among the priority hazardous substances within Directives 2000/60/EC (WFD) and 2008/105/EC. The Environmental Quality Standards (EQSs) set by the WFD for TBT are quite difficult to quantify by means of chemical analysis, without the use of expensive and high performance methods. Assuming that EQSs set for TBT were derived from evidence of imposex development at very low concentrations, this specific biomarker could be used as an indirect measure of assessing levels of bioavailable BTs. Therefore, this study aims to validate the use of imposex development as an investigative tool to monitor the bioavailable fraction of BTs within the WFD, by comparing imposex levels and BT concentrations in Nassarius nitidus from the Venice Lagoon. BT concentrations and imposex levels in N. nitidus, collected in 2013, had decreased when compared to previous studies in the same area. Both VDSI and RPLI correlated positively with BT body burden in females, confirming that imposex is a valid tool to monitor bioavailable BTs. However, TBT is still a matter of concern in the Venice Lagoon, as TBT concentrations were still higher than its degradation products suggesting recent fresh TBT inputs in the studied area. To propose imposex levels as an indicator of the impact of BTs within the WFD, classification class boundaries and Ecological Quality Ratios were introduced. As a preliminary attempt, imposex levels were also compared to the OSPAR Commission EcoQOs which linked imposex levels in Nassarius reticulatus with TBT concentrations in water. Based on this comparison the degree of imposex development in the Venice Lagoon suggested that TBT concentrations in water should be over the EQS-AA concentration and, at one site, also over the EQS-MAC. From all the results obtained in this work, it appears that imposex evaluation in N. reticulatus could give information about the ecological status regarding BT compounds under the WFD and also be used to monitor their effects and support chemical analyses until more sensitive methods become available.

Imposex levels and butyltin compounds (BTs) in Hexaplex trunculus (Linnaeus, 1758) from the northern Adriatic Sea (Italy): Ecological risk assessment before and after the ban.

The aim of this study was to compare imposex and butyltin compounds (BTs) data, collected before and after the organotin ban in 2008, in order to assess temporal and spatial variation of the phenomenon, the decline of BT contamination, and the effects on Hexaplex trunculus population in the coastal area of the northern Adriatic Sea, close to the Venice Lagoon. Both in marine and in lagoon sites, the results obtained in 2013-2015 showed a significant decline in the incidence of imposex in respect to those from the 2002 survey. In 2002, lagoon samples exhibited Relative Penis Size Index (RPSI) higher than marine samples, whereas no differences were detected in the recent survey, when all RPSI values were below 0.6%. Vas Deference Sequence Index (VDSI) mean values were over 4 before the ban introduction and below this value after that, indicating more critical conditions for gastropod population in 2002 rather than in 2013-15. Percentage of sterile females was up to 69% in 2002, whilst in the more recent survey no sterile female was found. Range of BT concentrations in gastropods decreased from 252 to 579 to 16-31ng∑BT/g d.w. BT body burdens varied according to a gender dependant pattern, with higher concentrations observed in females than in males. A first attempt to propose a classification based on BT impact on H. trunculus, according to the Water Framework Directive, revealed that most sites were in Bad ecological status before the ban and attained a Poor/Moderate status after that.

Organic aggregates formed by benthopleustophyte brown alga Acinetospora crinita (Acinetosporaceae, Ectocarpales).

This work presents the elemental, polysaccharide, and fatty acid compositions of benthic aggregates formed by the filamentous brown alga Acinetospora crinita, which are widely spread on the rocky bottoms of the Mediterranean Sea. The aggregates can be characterized as mineralized centers in which regeneration of nutrients and recycling of dissolved organic matter actively occur and favor the development of an abundant phytoplankton community. Analyses of the stable isotopes of C and N display their marine origin and could provide evidence of the processes that occur inside/outside of the aggregates. The monosaccharide compositions of Adriatic and Tyrrhenian mucilages produced by brown alga A. crinita were quite similar. In particular, the Adriatic sample compositions resembled the average composition of the Tyrrhenian high molecular weight exopolymers, and the observed differences could be ascribed to different degradation stages. The fatty acid patterns found for the aggregates were similar to those observed in the isolated A. crinita algae with variable contributions from embedded diatom species. The bacterial contribution to the fatty acid pool was quite low, most likely due to the known poor conditions for their heterotrophic growth.

Environmental quality assessment of Grand Harbour (Valletta, Maltese Islands): a case study of a busy harbour in the Central Mediterranean Sea.

Contamination levels by plastic debris, trace elements and persistent organic pollutants were assessed and related to macrobenthic diversity within soft bottoms of Grand Harbour (Malta, Central Mediterranean). Sediment toxicity was evaluated by ecotoxicological method, deploying Bacteria (Vibrio fischeri), Echinodermata (Paracentrotus lividus) and Crustacea (Corophium orientale). Univariate analysis (Pearson's test) was used to test relationships between biodiversity indices, pollutants and grain size. A multivariate approach (PERMANOVA) was applied to investigate for any significant differences among sampling stations concerning plastic abundances and to test the relationship between infaunal abundances and pollutant concentrations (the BIOENV test). Significant differences in the plastic abundances were found between sampling stations. The lowest value for Shannon-Wiener biodiversity index was associated to the highest sediment pollution level. Multivariate analyses suggest that MBT and TBT were factors that most influenced macrozoobenthic abundance and biodiversity. The bivalve Corbula gibba and the introduced polychaete Monticellina dorsobranchialis were the most abundant found species.

Evidence of butyltin biomagnification along the Northern Adriatic food-web (Mediterranean Sea) elucidated by stable isotope ratios.

The biomagnification of tributyltin (TBT), dibutyltin (DBT), monobutyltin (MBT), and total butyltins (ΣBT) was analyzed in the Northern Adriatic food-web (Mediterranean) considering trophodynamic interactions among species and carbon sources in the food-web. Although it is acknowledged that these contaminants bioaccumulate in marine organisms, it is still controversial whether they biomagnify along food-webs. A wide range of species was considered, from plankton feeders to top predators, whose trophic level (TL) was assessed measuring the biological enrichment of nitrogen stable isotopes (δ(15)N). Carbon isotopic signature (δ(13)C) was used to trace carbon sources in the food-web (terrestrial vs marine). At least one butyltin species was detected in the majority of samples, and TBT was the predominant contaminant. A significant positive relationship was found between TL and butyltin concentrations, implying food-web biomagnification. Coherently, the Trophic Magnification Factor resulted higher than 1, ranging between 3.88 for ΣBT and 4.62 for DBT. A negative but not significant correlation was instead found between δ(13)C and butyltin concentrations, indicating a slight decreasing gradient of contaminants concentrations in species according to the coastal influence as carbon source in their diet. However, trophodynamic mechanisms are likely more important factors in determining butyltin distribution in the Northern Adriatic food-web.

Tracing the route: Using stable isotope analysis to understand microplastic pathways through the pelagic-neritic food web of the Tyrrhenian Sea (Western Mediterranean).

This study provides a pilot investigation of the relationship between microplastic ingestion and the trophic ecology of three pelagic fish species (Engraulis encrasicolus, Scomber scombrus, and Trachurus trachurus) from Anzio coast, Tyrrhenian Sea (Western Mediterranean). Stable isotope analysis has been performed to determine the trophic position and the isotopic niche of the three species. Then, data on the occurrence, abundance, and diversity of ingested microplastics have been analyzed considering the observed foraging patterns. The detected differences in the estimated trophic position (E. encrasicolus = 3.08 ± 0.18; S. scombrus = 3.57 ± 0.21; T. trachurus = 4.07 ± 0.21), together with the absence of overlap in the isotopic niches confirm that the three examined species cover different ecological roles within the coastal-pelagic food web. Results from the analysis of ingested microplastics show that the trophic position has no remarkable effects on the incidence of microplastic ingestion, with no significant differences detected in terms of both frequency of occurrence and number of ingested microplastics per individual. However, differences among species emerge when considering the diversity of ingested microplastic types in terms of shape, size, color, and polymer composition. Species at higher trophic levels have shown to ingest a greater diversity of microplastics, including a significant increase in the size of the ingested particles (median surface area: 0.011 mm2 in E. encrasicolus; 0.021 mm2 in S. scombrus; 0.036 mm2 in T. trachurus). The ingestion of larger microplastics might be due to the larger gape sizes but also to active selection mechanisms, likely stimulated by the similarity of these particles to natural or potential prey of both S. scombrus and T. trachurus. Overall, this study suggests that microplastic ingestion can be affected by the different trophic position of fish species, providing new insights about the impact of microplastic contamination on the pelagic community.

First record of microplastic in the environmental matrices of a Mediterranean marine cave (Bue Marino, Sardinia, Italy).

This study investigates for the first time the presence of microplastics in sediment, water, and benthic organisms (foraminifera) of a marine cave in the Gulf of Orosei (Sardinia, Italy). Microplastics were found in all water, and sediment samples with similar shapes, sizes, and compositions; identified items were mainly fragments and fibers constituted by PVC and polyethylene. Their provenance was supposed to be predominantly from the sea than from the seasonal freshwater supplies from the karst system. Foraminiferal assemblages were mainly constituted by calcareous hyaline taxa in the outer station, while in the inner ones, the agglutinated Eggerelloides advenus was dominant. FTIR analyses on agglutinated shells identified polyethylene. Microplastic items are collected by the foraminifers and sediment grains building the shell chambers. This is the first study providing evidence that marine caves may be collectors of microplastics and that, in these habitats, microplastics enter the biotic matrix at the protist's level.

Snapshot of the Distribution and Biology of Alien Jellyfish Cassiopea andromeda (Forsskål, 1775) in a Mediterranean Touristic Harbour.

Harbors are hotspots for the introduction of alien species, and, usually, investigations on their host populations help fill the knowledge gap in their pathways of invasion and in their impacts on marine biodiversity and ecosystems. In 2014, the upside-down alien jellyfish Cassiopea andromeda invaded a Mediterranean touristic harbor ("Cala"), and its abundance has since increased over time. In the present study, the distribution and trophic behavior of C. andromeda in Cala were investigated for the years 2017-2018 through visual sampling, and GIS-based statistical and stable isotope analyses. Since Cala is a hard-to-reach area (with many anchor cables and boats), Megabenthos Underwater Video was used to count the number and estimate the size of jellyfishes. The variations in size throughout the study period suggest that the population of C. andromeda is quite established in Cala at depths lower than 7.5 m. The ranges of the environmental parameters recorded (temperature, salinity, and transparency) were consistent with the ideal conditions for maintaining a Cassiopea population, but they did not seem to influence aggregation. Additionally, the carbon and nitrogen isotopic signatures studied highlight the mixotrophic behavior of this species. These preliminary results confirm the capacity of C. andromeda to live and reproduce in heavily anthropized areas.

New method for simultaneous determination of dissolved organic carbon and its stable carbon isotope ratio in liquid samples: environmental applications.

This study reports the development of an all-in-one elemental analyser isotope ratio mass spectrometry (EA-IRMS) system modified for simultaneous analysis of dissolved organic carbon (DOC) concentration and its stable carbon isotope footprint (δ13CDOC) in aqueous samples. The method involves a quantitative oxidation of DOC in a 200 µL liquid sample to CO2, after sample acidification and stripping by nitrogen. The detection limit of the method for DOC quantification was 0.2 mg C/L with an analytical precision of 12 %. Uncertainty of stable isotope determinations was 2 % at 0.2 mg DOC/L, while decreasing to 0.3 % at 20 mg DOC/L. Quantitative oxidation of DOC in aqueous samples was validated by using ring test water samples and Deep Sea reference seawater. The method performances of isotope analysis were evaluated by analysing different isotopic standard solutions. The applicability of the method was tested through the analysis of different environmental types of water, showing that δ13CDOC ranged from - 23.30 to -31.85 ‰, allowing to characterize samples of different environmental origin. The developed method offers several advantages including rapidity, use of small sample volumes and minimal sample pre-treatment, making it a valuable tool for routine DOC concentration measurements paired with isotopic characterization.

Environmental restoration by aquatic angiosperm transplants in transitional water systems: The Venice Lagoon as a case study.

The paper reports the results obtained after 4 years of aquatic angiosperm transplants in areas of the Venice Lagoon (North Adriatic Sea, Mediterranean) where meadows almost disappeared due to eutrophication, pollution and overexploitation of clam resources. The project LIFE12 NAT/IT/000331-SeResto, funded by the European Union, allowed to recolonize the Habitat 1150* (coastal lagoons) in the northernmost part of the lagoon, by extensive manual transplants of small sods or single rhizomes of Zostera marina, Zostera noltei, Ruppia cirrhosa and, in some stations also of Cymodocea nodosa. Over the 4 years of the project more than 75,000 rhizomes were transplanted in 35 stations with the support of local stakeholders (fishermen, hunters and sport clubs). Plants took root in 32 stations forming extensive meadows on a surface of approx. 10 km2 even if some failures were recorded in areas affected by outflows of freshwater rich in nutrients and suspended particulate matter. The rapid recovery of the ecological status of the involved areas was the result of this meadow restoration, which was in compliance with Water Framework Directive (WFD 2000/60/EC) objectives. Moreover, the monitoring of environmental parameters in the water column and in surface sediments allowed to identify the best conditions for successful transplants. Small, widespread interventions and the participation of local stakeholders in the environmental recovery, make this action economically cheap and easily transposable in other similar environments.

Preliminary multi analytical approach to address geographic traceability at the intraspecific level in Scombridae family.

Globalization of seafood product marketing caused the increase of request of an effective fish traceability that enhances the consumer confidence in food safety. In this study, an integrated multi analytical approach based on two different and independent analytical techniques (carbon and nitrogen stable isotopes and fatty acids analysis) was applied in order to identify different fish species and trace their geographical provenience. The investigation was focused on four species (Thunnus thynnus, Thunnus alalunga, Auxis rochei and Scomber scombrus) belonging to the Scombridae family. The DNA barcoding method confirmed genus and species for S. scombrus and A. rochei, but only genus for T. alalunga and T. thynnus. Carbon and nitrogen stable isotopes results evidenced different fish diets and trophic positions, whereas fatty acids analysis displayed that the unsaturated prevailed (∼60 %) over the saturated compounds with a variation among the species and the geographical area in particular for docosahexaenoic and eicosapentaenoic acids percentage. The principal component analysis applied to stable isotopes and fatty acids evidenced a good discrimination among species and their geographical catching area. This multi-disciplinary analytical approach could represent a promising tool to identify the commercial fish and trace their origin in order to guarantee the health of consumers.

Common patterns of functional and biotic indices in response to multiple stressors in marine harbours ecosystems.

Evaluating the effects of anthropogenic pressure on the marine environment is one of the focal objectives in identifying strategies for its use, conservation and restoration. In this paper, we assessed the effects of chemical pollutants, grain size and plastic litter on functional traits, biodiversity and biotic indices. The study was conducted on the benthic communities of three harbours in the central Mediterranean Sea: Malta, Augusta and Syracuse, subjected to different levels of anthropogenic stress (high, medium and low, respectively). Six traits were considered, subdivided into 22 categories: reproductive frequency, environmental position, mobility, life habit, feeding habit and bioturbation. Functional diversity indices analysed were: Functional Divergence, Quadratic Entropy, Functional Evenness and Functional Richness. To assess the trait responses to environmental gradients, we applied RLQ analysis, which considers simultaneously the relationship between three components: environmental data (R), species abundances (L) and species traits (Q). From our analyses, significant relationships (P-value = 0.0018 for permutation of samples, and P-value = 0.00027 for permutation of species) between functional traits and environmental data were highlighted. The trait categories significantly influenced by environmental variables were those representing feeding habits and mobility. In particular, the first category was influenced by chemical pollutants (organotin compounds and polycyclic aromatic hydrocarbons) and grain size (silt and sand), while the latter category was influenced only by chemical pollutants. Pearson correlations performed for functional vs biotic and diversity indices confirmed the validity of the chosen conceptual framework for harbour environments. Finally, linear models assessing the influence of stressors on functional parameters underlined the link between environmental data vs benthic and functional indices. Our results highlight the fact that functional trait analysis provides a useful and fast method for detecting in greater depth the effects of multiple stressors on functional diversity in marine ecosystems.

Organotin compounds in touristic marinas of the northern Adriatic Sea: occurrence, speciation and potential recycling at the sediment-water interface.

Butyltin compound (BTC) contamination was evaluated in two north Adriatic marinas, San Rocco (Italy) and Lucija (Slovenia). BTC sedimentary concentrations (121 ± 46 and 352 ± 30 ng Sn g-1 in San Rocco and Lucija, respectively) evidenced the past use of antifouling paints, confirmed by the reduced tributyltin content (~ 46%) with respect to the sum of BTC. Elemental and organic carbon isotopic (δ13C) analyses of bulk sediments and its lipid and humic substances were performed in order to evaluate their role in BTC partitioning and preservation. The δ13C of sedimentary bulk and refractory organic matter suggested that diagenetic processes could play a role in the preservation or release of pollutants. No contamination was found in water collected from the benthic chamber and thus, fluxes at the sediment-water interface were not assessed, except for MBT efflux at Lucija (28.9 ng Sn m-2 day-1). Nevertheless, BTC concentrations in porewaters (up to 75 ng Sn l-1) and rather low sediment-porewater partitioning coefficients (Kd) with respect to the data reported in the literature would suggest a potential risk of the reintroduction of BTC into the water column at both sites: at Lucija, sedimentary contamination is high despite the greater Log Kd, whilst at San Rocco, the low BTC concentration is associated with a reduced sediment affinity.

Meiofaunal communities in four Adriatic ports: Baseline data for risk assessment in ballast water management.

Ports receive a variety of contaminants related to a wide range of anthropogenic activities - including ship ballast water (BW) - that ultimately find their way to sediments. Benthic meiofauna from four Adriatic ports (Ancona, Trieste, Koper, and Split) was assessed for the main environmental pollutants, to evaluate the effects of human activities on meiobenthos and identify the most appropriate descriptor to assess the ecological quality of marine ecosystems. Sediment analysis demonstrated that Trieste and Split were the most contaminated ports, followed by Koper and Ancona. All meiofaunal parameters showed high spatial and temporal variability, in line with the marked heterogeneity of the four ports. Sand, total organic carbon, and pollutants seemed to be the variables that best explained meiofaunal patterns. Community structure and rare taxa were the meiofaunal descriptors that reflected the environmental status and biological response most accurately. The present data suggest that meiofauna can be used to assess the biological impact of BW.

Contribution of deltaic wetland food sources to coastal macrobenthic consumers (Po River Delta, north Adriatic Sea).

Estuaries are known to export huge quantities of terrigenous/riverine organic matter and nutrients to coastal areas, subsidizing food sources to their communities; however, this process is seldom investigated in complex multichannel delta-prodelta systems. Stable isotope analysis was used to investigate the contribution of organic sources originated from the Po River Delta, one of the widest deltaic wetlands in the northern Mediterranean Sea, to the diet of the nearby coastal macrobenthic consumers, and their influence on invertebrates' trophic structure and biomass. Following intense river flood events, macrofauna samples were collected in the prodelta area at increasing distance from the main river distributary mouth. Potential primary resources were collected within the delta and the prodelta areas. A terrestrial signal in the primary consumers' diet was evident as a shift in their δ13C, being this more 13C depleted near the main river distributary. The Bayesian mixing models indicated an important contribution of deltaic sources, in particular C4-grass detritus (25-57%), to primary consumers' diet, confirming a strong energetic link between delta and prodelta areas. Overall, most of the consumers' biomass were concentrated at sites close to the main distributary mouth, mainly as suspension and surface deposit feeders. A simplification of the macrobenthic community structure, accompanied by narrower transfer pathways was also evident at sites more distant from the main river distributary, in relation to changes in the quality of resources. Our data, although limited to winter season, suggest that during periods of low in situ productivity but high river flow energy, invertebrates are able to efficiently exploit terrigenous food sources, restraining the detrimental effect of increased turbidity and sedimentation. We conclude that riparian/wetland vegetation associated with river deltas can provide important food sources to marine primary consumers. These results furthermore highlight the need for integrated management and protection strategies of connected land-sea ecosystems.

Relationships between plastic litter and chemical pollutants on benthic biodiversity.

Five Descriptors (D) of Marine Strategy Framework Directive (MSFD): marine litter (D10), non-indigenous species (D2) and organic and inorganic pollutants (D8), were estimated in a coastal area of GSA 16 (Augusta harbour, Central Mediterranean Sea) in order to study their effects on the biodiversity (D1) of the benthic community D6) and to improve data for the MSFD. Investigation of plastic debris had led to the identification of 38 fragments divided into four categories, among which microplastics resulted as the most abundant. Six non-indigenous species, belonging to Polychaeta (Kirkegaardia dorsobranchialis, Notomastus aberans, Pista unibranchia, Pseudonereis anomala, Branchiomma bairdi) and Mollusca (Brachidontes pharaonis) were found. Biodiversity and benthic indices suggested a generalised, slightly disturbed ecological status. Anthracene, Zinc and Chrome were the most abundant chemical compounds in analysed sediments. Significant correlations were found between the abundance of trace elements vs biotic indices and between plastic debris vs biodiversity and benthic indices. This study represents the first report about the abundance of plastic debris and its relationship to contaminants and infauna in Augusta harbour. Our results can provide useful information for national and international laws and directives.

Multidisciplinary studies on a sick-leader syndrome-associated mass stranding of sperm whales (Physeter macrocephalus) along the Adriatic coast of Italy.

Mass strandings of sperm whales (Physeter macrocephalus) are rare in the Mediterranean Sea. Nevertheless, in 2014 a pod of 7 specimens stranded alive along the Italian coast of the Central Adriatic Sea: 3 individuals died on the beach after a few hours due to internal damages induced by prolonged recumbency; the remaining 4 whales were refloated after great efforts. All the dead animals were genetically related females; one was pregnant. All the animals were infected by dolphin morbillivirus (DMV) and the pregnant whale was also affected by a severe nephropathy due to a large kidney stone. Other analyses ruled out other possible relevant factors related to weather conditions or human activities. The results of multidisciplinary post-mortem analyses revealed that the 7 sperm whales entered the Adriatic Sea encountering adverse weather conditions and then kept heading northward following the pregnant but sick leader of the pod, thereby reaching the stranding site. DMV infection most likely played a crucial role in impairing the health condition and orientation abilities of the whales. They did not steer back towards deeper waters, but eventually stranded along the Central Adriatic Sea coastline, a real trap for sperm whales.

Preliminary study to characterize plastic polymers using elemental analyser/isotope ratio mass spectrometry (EA/IRMS).

Plastic waste is a growing global environmental problem, particularly in the marine ecosystems, in consideration of its persistence. The monitoring of the plastic waste has become a global issue, as reported by several surveillance guidelines proposed by Regional Sea Conventions (OSPAR, UNEP) and appointed by the EU Marine Strategy Framework Directive. Policy responses to plastic waste vary at many levels, ranging from beach clean-up to bans on the commercialization of plastic bags and to Regional Plans for waste management and recycling. Moreover, in recent years, the production of plant-derived biodegradable plastic polymers has assumed increasing importance. This study reports the first preliminary characterization of carbon stable isotopes (δ13C) of different plastic polymers (petroleum- and plant-derived) in order to increase the dataset of isotopic values as a tool for further investigation in different fields of polymers research as well as in the marine environment surveillance. The δ13C values determined in different packaging for food uses reflect the plant origin of "BIO" materials, whereas the recycled plastic materials displayed a δ13C signatures between plant- and petroleum-derived polymers source. In a preliminary estimation, the different colours of plastic did not affect the variability of δ13C values, whereas the abiotic and biotic degradation processes that occurred in the plastic materials collected on beaches and in seawater, showed less negative δ13C values. A preliminary experimental field test confirmed these results. The advantages offered by isotope ratio mass spectrometry with respect to other analytical methods used to characterize the composition of plastic polymers are: high sensitivity, small amount of material required, rapidity of analysis, low cost and no limitation in black/dark samples compared with spectroscopic analysis.

DoE optimization of a mercury isotope ratio determination method for environmental studies.

By using the experimental design (DoE) technique, we optimized an analytical method for the determination of mercury isotope ratios by means of cold-vapor multicollector ICP-MS (CV-MC-ICP-MS) to provide absolute Hg isotopic ratio measurements with a suitable internal precision. By running 32 experiments, the influence of mercury and thallium internal standard concentrations, total measuring time and sample flow rate was evaluated. Method was optimized varying Hg concentration between 2 and 20 ng g(-1). The model finds out some correlations within the parameters affect the measurements precision and predicts suitable sample measurement precisions for Hg concentrations from 5 ng g(-1) Hg upwards. The method was successfully applied to samples of Manila clams (Ruditapes philippinarum) coming from the Marano and Grado lagoon (NE Italy), a coastal environment affected by long term mercury contamination mainly due to mining activity. Results show different extents of both mass dependent fractionation (MDF) and mass independent fractionation (MIF) phenomena in clams according to their size and sampling sites in the lagoon. The method is fit for determinations on real samples, allowing for the use of Hg isotopic ratios to study mercury biogeochemical cycles in complex ecosystems.

Spectroscopic evidence of the marine origin of mucilages in the Northern Adriatic Sea.

Vibrational Fourier transform infrared (FTIR) and UV-Visible spectroscopies coupled to thermogravimetry were used to characterise the gelatinous aggregates as well as the humin and humic fractions extracted from the aggregates. Spectroscopic analysis made it possible to identify the main classes of compounds present in the samples. The FTIR spectra of the aggregates showed the presence of carbohydrates, saturated aliphatic compounds, aminic, esteric, amidic (proteins) and ketonic compounds, organic and inorganic phosphates, silica and, in some cases, carbonates. The FTIR spectra of humin and humic acids showed functional groups similar to those found in the spectra of the aggregates. The high aliphatic character and the absence of aromatic compounds observed in many aggregates sampled far from estuarine areas suggest the marine origin of mucilages. These results were also confirmed by the 270/407 nm (A2/A4) and 465/665 nm (E4/E6) absorbance ratio in humic acids.