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Twenty-five species of cetaceans have been reported throughout the Mediterranean Sea, eight of them are commonly distributed in the whole basin and are regularly found beached or adrift in the sea. Stranded animals are frequently found in poor conservation status, preventing reliable identification; identification is thus often based solely on morphological features. Therewith, molecular tools are especially useful to provide taxonomic identification. In this work, a four-enzymes PCR-RFLP in silico protocol, based on a fragment of the mitochondrial gene cytb, has been designed for cetacean species occurring in the Mediterranean Sea. Moreover, beached or floating specimen samples belonging to the eight common species have been tested in the laboratory, providing evidence that this approach represents a reliable, cost- and time-effective tool for their specific identification.

Marine bioinvasions and other rapid biodiversity changes require today integrating existing monitoring tools with other complementary detection strategies to provide a more efficient management. Here we explored the efficacy of fishermen observations and traditional port surveys to effectively track the occurrence of both indigenous and non-indigenous megafauna in the Adriatic Sea. This consisted mainly of mobile taxa such as fishes, crustaceans and molluscs. Port surveys using traps and nets within 10 major Adriatic harbours, were compared with the information obtained from 153 interviews with local fishermen. Information gathered by traps and nets varied significantly and generally resulted of limited efficacy in exotic species detection. Interviews allowed tracking the occurrence of new species through time and space, providing complementary knowledge at the low cost. This combined approach improves our capability of being informed on the arrival of species of different origin, providing a more rational, improved basis for environmental management and decision making.

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.

The accumulation of Posidonia oceanica dead leaves on the beaches of the Mediterranean shores is a natural phenomenon. They are either temporary or permanent structures (banquettes) and represent a valuable resource, with important ecosystem functions including coastal protection against erosion. Nevertheless, the perception of these plant accumulations by the different stakeholders (beach managers, local administrations and tourists) is often negative, they consider these deposits a malevolent waste to be removed, rather than a natural and valuable component of the coastline. We propose an integrated/beneficial management model for posidonia deposits, called ECOLOGICAL BEACH, firstly proposed in France, and recently implemented and applied in Italy. The model promotes the preservation of posidonia beach casts on site, with a balanced coexistence of natural and anthropic elements. The model fosters the several important ecosystem services of the beach casts and contributes to coastal preservation. To successfully spread the model, several activities must be implemented: a regulatory framework, the collection of data about the occurrence of beach casts, management protocols and educational programs. The most important activity is the educational one, based on the dissemination of the ecological and economic value of the beach casts, aimed at switching the perception of this phenomenon towards positive appraisal.

The MYTIAD project focused on the assessment of chemical contamination (metals, PAHs, PCBs and organochlorine pesticides) in the Adriatic coastal waters by active mussel watching. Its purpose was harmonising and standardising strategies and methodologies used to assess the contamination status of the Adriatic Sea, in the framework of the WFD, the MSFD and UNEP/MAP Integrated Monitoring and Assessment Programme. Data on metals pointed out some hotspots along the eastern and western Adriatic Sea coasts, with the highest values of total PAH concentration detected in Split, Trieste, and Taranto (Ionian Sea). PCB and endrin contamination was higher in the Adriatic Sea than in the western Mediterranean Sea. This is the first comprehensive overview of contamination in the Adriatic Sea with critical comparisons of related studies over the Mediterranean Sea. It provides a useful harmonised dataset to support a coordinated definition of baselines, targets and thresholds, and further management of chemical contamination.

Oxidant treatment of ballast water (BW) is commonly used in BW systems in order to minimize the transport of alien species. The release of disinfection by-products (DBPs) associated to the treatment of BW and cross-contamination of butyltin (BT) compounds through BW discharge is a topic of environmental concern. A chemical port baseline survey has been conducted in seven ports of the Adriatic Sea. Analysis have been performed on transplanted mussels, surface sediment, seawater, BW. Results showed an evidence of BT contamination, particularly in sediments, probably related to their illegal usage or to intensive shipping activities. Therefore, BW may act as a vector and contribute to re-buildup of BT contamination in the coastal regions. A baseline set of data concerning DBPs is provided, showing the preferential distribution of these compounds in the marine environment that will be useful for future considerations on monitoring and assessment of chemical contamination associated with BW.

The MYTILOS project aimed at drawing up a preliminary report on coastal chemical contamination at the scale of the Western Mediterranean (continental coasts of the Balearic Islands, Sicily, Sardinia, Corsica and Maghreb) based on a transplanted mussels methodology validated along the French coasts since 1996 by Ifremer and the Rhône Méditerranée & Corsica water board. MYTILOS is backed up by the INTERREG III B/MEDOC programme, the PNUE/PAM-MEDPOL and Rhône Méditerranée & Corsica water board. Three cruises (2004, 2005, 2006) have taken place to assess the first state of chemical contamination along the Western Mediterranean shores with the same methodology. Approximately 120 days were spent at sea deploying and retrieving 123 mussel bags. The results obtained for all studied contaminants were equivalent to those obtained along the French coast according the RINBIO network. These similarities relate to both the highest measured levels and background levels throughout the 123 stations. The areas of greatest impact were mainly urban and industrial centers and the outlets of major rivers, with a far higher midsea impact on the dilution of organic compounds than on metals. Metal levels measured in midsea zones were found to be similar to those in natural shellfish populations living along the coast. On a global scale we can observe that the contaminants levels in the Mediterranean Sea are in the same range as in other areas worldwide. Overall, the research demonstrates the reliability of this methodology for marine pollution monitoring, especially in the Mediterranean sea. © 2010 Springer Science+Business Media, LLC.

The Mytilos project was carried during the last 3 years (2004-2006) in order to evaluate the level of chemical contamination along the coasts of the western basin of the Mediterranean Sea. Active mussel watch consisted of caged (12 weeks) Mytilus galloprovincialis at 123 stations along the Mediterranean coasts of Spain, France, Italy, North Tunisia, Algeria and Morocco. Results indicated total PAHs levels in the range 22-106 μg/kg dry weight. All large towns or industrial areas from all countries of the basin were affected. Analysis of the origin of contamination demonstrated mainly a pyrolytic origin except in some areas such as Maddalena, Napoli, Alger, and Cornigliano (Genova). Statistical analysis of the PAHs accumulation data enabled the characterisation and ranking of each coastal area. The overall results give scientific and technical basis for large-scale monitoring of the PAH contamination in the entire western Mediterranean basin. © 2010 Springer Science+Business Media B.V., This research was supported by EEC (Interreg/medoccIIIC/MYTILOS). INRH (Morocco), ISMAL (Algeria) and INSTM (Tunisia) were supported by UNEP MEDPOL.

The signaling pathways involved in mussel immune defence were investigated utilizing a model of killing of Escherichia coli by Mytilus galloprovincialis hemocytes in a co-culture setting. In particular, the role played by different mitogen activated protein kinases (MAPKs) and by the production of eicosanoids were investigated utilising specific cell permeant, pharmacological enzyme inhibitors. Hemocyte pretreatment with the p38 MAPK inhibitor SB203580 significantly reduced bacterial killing, whereas PD98059 (an inhibitor of ERK--extracellularly regulated kinase--MAPK activation) had no significant effect. Wortmannin also inhibited bacterial killing, indicating a crucial role for PI3-kinase activation in the immune response. Killing of E. coli was also reduced by inhibitors of both PLA2 and cyclooxygenase activities, indicating that eicosanoid production is involved in mediating the response to bacterial challenge. The results demonstrate that bacterial killing by mussel hemocytes is particularly sensitive to inhibitors of the key steps involved in the transduction of bacterial signals into the host cell. Moreover, these data indicate that the hemocyte bactericidal activity can be suitably utilized not only for identifying the signaling pathways involved in the response to bacterial infection, but also as a potential investigative-toxicology model to test drugs and contaminants for their effect on the overall mussel immune defence.

Autores: Benedicto, José ... [et al.]. -- 11 páginas, 3 figuras, 1 tabla, A large scale study of trace metal contamination (Hg, Cd, Pb and Ni) by means of caged mussels Mytilus galloprovincialis was undertaken along the coastal waters of the Western Mediterranean Sea within the context of the MYTILOS project. Individual mussels from an homogeneous population (shell size 50 ± 5 mm) obtained from an aquaculture farm were consecutively caged and deployed at 123 sites located in the Alborán, North-Western, South-Western and Tyrrhenian sub-basins for 12 weeks (April–July) in 2004, 2005 and 2006. After cage recoveries, both the metal content in the whole mussel tissue and the allometric parameters were measured. Statistical analysis of the datasets showed significant differences in concentrations between sub-basins for some metals and mussel condition index (CI). Linear regression models coupled to the CI were revisited for the data adjustment of certain trace metals (Hg, Cd and Ni), and four level categories were statistically derived to facilitate interregional comparison. Seawater masses surrounding coastal areas impacted by run-off from land mineralised coasts and industrial activities displayed the highest concentration ranges (Hg: 0.15–0.31 mg kg−1 dw; Cd: 1.97–2.11; Ni: 2.18–3.20 and Pb: 3.1–3.8), although the levels obtained in most of the sites fitted within moderate or low categories, and they could be considered as baseline concentrations. However, few sites considered little-influenced by human activities, at present, showed high concentrations of Cd, Ni and Pb, which constitute new areas of concern. Overall, the use of active biomonitoring (ABM) approach allowed to investigate trace metal contamination in order to support policy makers in establishing regional strategies (particularly, with regard to the European Marine Strategy Directive)., Esta investigación ha sido financiada por el EEC (Proyecto Interreg/MedoccIIIB/MYTILOS). El Programa UNEP MEDPOL ha subvencionado a: INRH (Marruecos), ESSMAL (Algeria) e INSTM (Túnez).

8 pages, 3 figures, 3 tables, The effects of sewage outfalls on subtidal fish assemblages were studied along the NW coasts of Malta (Sicily channel, Mediterranean Sea) by means of underwater visual census. The presence of two spatially distinct outfalls discharging untreated wastewaters allowed to use a balanced symmetrical after control/impact (ACI) design that consisted of two putatively impacted locations and two controls, with four sites nested in each location. Surveys were performed in 2006 at two random dates. The study highlighted significant changes at both assemblage and individual species levels. Fish assemblages structures were different between controls and sewages, where total abundance of fish were higher. The responses of individual species to sewage pollution were mostly related to an anomalous increase of two small opportunistic species i.e. Gobius bucchichii and Parablennius rouxi and to a decrease of species of the genus Symphodus, particularly S. roissali and S. ocellatus. Moreover in correspondence of the outfalls, significant changes of the fish size distribution were detected for several species. These results support the use of fish assemblages as biological indicators for marine coastal waters and demonstrated the possibility to obtain sharp signals of environmental impact from some individual fish species, This research was funded by the Interreg IIIA Italia-Malta Project

Autores: Scarpato, Alfonso ... [et al.]. -- 12 páginas, 7 figuras, 1 tabla, In order to evaluate the contamination levels in the Western Mediterranean basin, the active mussel watch methodology has been applied. This methodology consists of mussel transplantation (Mytilus galloprovincialis) from non impacted areas to selected coastal areas, characterised by potential impact from the continent due to contaminating sources. The areas of interest were selected along the entire coastal development of the Western Mediterranean sea, 122 sites in total. The time of mussel caging exposure was 12 weeks. The project was co-financed in the frame of the Interreg IIIB Meddoc Programme, aimed at determining the overall chemical quality of the Mediterranean sea, consistent with the Water Framework Directive 2000/60. Several partners representative of the coastal Mediterranean Countries were involved in the Project, with the purpose of building up a common surveillance network, adopting shared methodologies. In this paper we present the results of three yearly monitoring campaigns (2004, 2005, 2006) carried out along the coasts of Italy, France, Spain, Morocco, Algeria and Tunisia, including the coastal environment of Baleares, Sicily, Sardinia and Corsica. The contamination levels of Pesticides (DDT and its metabolites, Hexachlorocyclohexane isomers α and γ) and Polychlorinated biphenyls, are reported and discussed. Statistical elaborations performed on the original data set were mainly aimed at validating the raw sample distributions, by means of the Johnson method. Both DD and PCB species frequency distributions have been approximated to appropriate theoretical distributions, belonging to the Log-normal and Bounded families. By integrating the related Probability Density Functions (p.d.f.), different accumulation values for DDT, DDD and DDE and PCB species have been estimated, corresponding to fixed percentage points of the area under the respective curves. By choosing appropriate probability level boundaries (33rd and 66th percentile), different regional zones have been ranked in terms of low, medium and high accumulation for Pesticides and PCBs., Esta investigación ha sido financiada por la Comunidad Económica Europea (EEC) (Interreg/ MedoccIIIC). Marruecos, Algeria y Túnez fueron financiados por UNEP MEDPOL.

The common mussel Mytilus galloprovincialis was selected as unique biomonitor species to implement a regional monitoring programme, the CIESM Mediterranean Mussel Watch (MMW), in the Mediterranean and Black Seas. As of today, and upon standardization of the methodological approach, the MMW Network has been able to quantify Cs-137 levels in mussels from 60 coastal stations and to produce the first distribution map of this artificial radionuclide at the scale of the entire Mediterranean and Black Seas. While: measured Cs-137 levels were found to be very low (usually

Polychlorinated biphenyls (PCBs) are industrial chemicals which have been released into the environment resulting in widespread and persistent contamination. PCBs exist as 209 different congeners depending on the chlorine substitution on the biphenyl rings; the physical properties and the toxic effects of a PCB congener are structure-dependent. In this work, individual ortho-substituted non coplanar PCB congeners were tested for their effects on the function of mussel (Mytilus galloprovincialis Lam.) hemocytes. Moreover, the possibility that in mussel hemocytes different PCBs may affect the signal transduction pathways involved in the immune response was investigated, with particular regards to relevant components of tyrosine-kinase mediated cell signaling. The results were compared with those obtained with a model of non-ortho-substituted coplanar congener. The results demonstrate that the di-ortho-substituted, non coplanar PCB congeners P47 (2,2',4,4'-tetrachlorobiphenyl) and P153 (2,2',4,4',5,5'-hexachlorobiphenyl) can alter immune parameters of mussel hemocytes, such as microbicidal activity and lysosomal enzyme release, respectively. Both congeners, as well as the non-ortho, coplanar congener P77 (3,3',4,4'-tetrachlorobiphenyl) significantly reduced hemocyte lysosomal membrane stability; however, P77 had no effect on either bacterial killing or lysozyme release. P47, P153 and P77 affected different components of tyrosine kinase-mediated cell signalling; in particular, they lead to a time-dependent increase in the phosphorylation level of the stress activated p38 and JNK Mitogen Activated Protein Kinases (MAPKs), as evaluated by Western blotting of hemocyte protein extracts with specific anti-phospho-MAPK antibodies. P153 also increased the level of phosphorylated ERK (extracellularly regulated) MAPKs. Moreover, non coplanar P47 and P153 caused increased tyrosine phosphorylation of the transcription factor STAT5, thus possibly affecting gene expression, whereas coplanar P77 was ineffective. The results demonstrate that MAPKs, and in particular the stress-activated p38 and JNK MAPKs, that represents a key step in the response of mussel hemocytes to bacterial infection, are a target for different non coplanar and coplanar PCB congeners. The results also show functional differences between different PCB congeners with respect to the hemocyte functions. However, chlorine substitution at the ortho positions is not necessarily related to immunotoxicity: the hexachlorinated P128 (2,2',3,3',4,4'-hexachlorobiphenyl) had no significant effect on mussel hemocytes, whereas its isomer P153, that represents a major component of environmental PCBs, and that is accumulated in mussel tissues, significantly affected both aspects of the immune response and relevant signal transduction pathways. These are the first data on the effects and possible mechanisms of immunotoxicity of non coplanar PCBs in mussel hemocytes. The results support the hypothesis that the innate immune system is a sensitive target for these contaminants in both vertebrates and invertebrates. Moreover, when considering that non coplanar congeners are present both in commercial mixtures and, in higher proportions, in environmental samples, the results suggest that bivalve hemocytes represent a useful model for evaluating the potential immunotoxicity of PCB contamination.

In this work the mechanisms of transduction triggered in Mytilus galloprovincialis hemocytes by bacterial challenge were investigated in an in vitro model of infection of hemocyte monolayers with Escherichia coli. Western blot analyses of hemocyte extracts with phospho-specific anti-MAPK (Mitogen Activated Protein Kinase) antibodies indicate that E. coli induced a time dependent activation of different classes of MAPKs, mainly of the stress-activated p38 MAPK. P38 activation was confirmed by the use of the selective kinase inhibitor SB203580. Moreover, hemocyte pretreatment with SB203580 significantly reduced bacterial killing, whereas PD98059, an inhibitor of extracellularly regulated kinase (ERK) activation, was ineffective. Interestingly, the PI3-kinase (phosphatidylinositol-3-OH-kinase) inhibitor, Wortmannin, reduced both p38 activation and bacterial killing, indicating a critical role also for this lipid kinase in the hemocyte immune response.