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In this study, the abundance and properties (size, shape, and polymer type) of microplastics (MPs) in sea surface water samples, collected during two sampling campaigns over 2018–2019, in four coastal areas of the Mediterranean Sea (Saronikos Gulf, LIgurian Sea, Gulf of Lion, and Gabes Gulf) were investigated. Coupled hydrodynamic/particle drift model simulations with basin-scale Mediterranean and high resolution nested models were used to provide a better understanding on the variability of the abundance/size of MPs, originating from wastewater and river runoff, in the four areas. Different size classes of MPs were considered in the model, taking into account biofouling induced sinking, as a possible mechanism of MPs removal from the surface. The Gabes Gulf showed the highest mean MPs abundance (0.073–0.310 items/m2), followed by Ligurian Sea (0.061–0.134 items/m2), Saronikos Gulf (0.047–0.080 items/m2), and Gulf of Lion (0.029–0.032 items/m2). Overall, the observed MPs abundance and size distribution was reasonably well reproduced by the model in the four different areas, except an overestimation of small size contribution in Saronikos Gulf. The basin-scale simulation revealed a strong decrease of smaller size MPs in offshore areas, due to biofouling induced sinking, with larger (floating) MPs being able to travel longer distances in the open sea. A significant impact of waves drift and advection of MPs from non-local sources was identified from model simulations, particularly in the Gulfs of Lion and Gabes, having a stronger effect on larger microplastics. In Gabes Gulf, most MPs originated from offshore areas, being mainly (floating) larger size classes, as suggested by the observed quite small contribution of

Component: C 1.3 Support for Member States' implementation of Maritime Spatial Planning Sub‐component: C 1.3.6. Establish Case Studies on Approaches to MSP Implementation Deliverable: Tyrrhenian Case Study The Tyrrhenian case study comprehends portions of internal, territorial and international waters and the entire study area is included within the Specially Protected Area of Mediterranean Importance (SPAMI) of the Pelagos Sanctuary for the Conservation of Marine Mammals, a transboundary Sanctuary established through an international agreement signed between three different countries (Italy, France and Principality of Monaco). It includes key areas in the context of Western Mediterranean in terms of ecological and socio-economic value, in particular three Italian National Parks, Asinara Island, La Maddalena Archipelago and the whole Tuscan Archipelago; the Corsica Channel, an area characterised by a great environmental sensitivity, subject to increased anthropic pressure for which the Italian and French Governments have signed a MoU concerning the regulation of international maritime traffic; the Bonifacio Strait, an international strait shared between French and Italian jurisdictions; and the only Particularly Sensitive Sea Area declared by the International Maritime Organization (IMO) in the Mediterranean Sea. Several human activities take place and interact dynamically with local uses and the marine environment components. Among these activities some are expected to increase in coming years. Thus, it will be fundamental to effectively organize their spatial distribution and mutual interactions in balance with the protection and sustainable management of the natural resources, taking in consideration the priority of avoiding impacts on marine mammals due to their great conservation relevance. In order to address specific details of the SIMWESTMED Tyrrhenian case study, the analysis process was directed through two main foci: a thematic focus (in the whole area) on the processes and instruments for the transboundary cooperation on MSP, coordinated by UNEP PAP/RAC; and a management focus on the Tuscan Archipelago, coordinated by CORILA. To address the thematic focus in the whole area, processes and instruments for transboundary cooperation on MSP were exhaustively and thoroughly analysed in depth, at different levels (international, EU, national), together with the different governance levels and requirements for a comprehensive application of MSP. In particular, the relevant legal framework and/or concerning the formal planning systems, and cross-border cooperation processes, governance context, relevant for the particularities of preservation of marine mammals as well as existing institutional regional cooperation structures were assessed. Based on these, the proposals on the MSP approach, addressing the appropriate measures in order to achieve greater protection of marine mammals in the Mediterranean, were developed., This report/document was produced as part of the SIMWESTMED Project (Grant Agreement N0. EASME/EMFF/2015/1.2.1.3/02/SI2.742101). PROJECT: Supporting Implementation of Maritime Spatial Planning in the Western Mediterranean region (SIMWESTMED) Competition for maritime space – for renewable energy equipment, aquaculture and other uses – has highlighted the need to manage our waters more coherently. Maritime spatial planning (MSP) works across borders and sectors to ensure human activities at sea take place in an efficient, safe and sustainable way. That is why the European Parliament and the Council have adopted a legislation to create a common framework for maritime spatial planning in Europe. The Directive 2014/89/EU of the European Parliament and of the Council of 23 July 2014 (said Maritime Spatial Planning Directive) establishes a framework in order to reduce conflicts between sectors and create synergies between different activities, to encourage investment – by creating predictability, transparency and clearer rules, to increase cross-border cooperation – between EU countries to develop energy grids, shipping lanes, pipelines, submarine cables and other activities, but also to develop coherent networks of protected areas, and to protect the environment – through early identification of impact and opportunities for multiple use of space. The SIMWESTMED project (Supporting Implementation of Maritime Spatial Planning in the Western Mediterranean region) is an EU/DG Mare co-funded cross-border project. It was launched on 1st of January 2017 and involves Spain, France, Italy and Malta, while these countries had just designated their Competent Authorities and transposed the Maritime Spatial Planning (MSP) Directive. SIMWESTMED aims to support the implementation of the MSP Directive in the waters of Spain, France, Italy and Malta, as well as to establish cross-border cooperation mechanisms between these Member States, to contribute to the coherence of their marine spatial plans to be established by 2021. The action ran until 31st of December 2018 and was based on a partnership of public bodies of the countries and two international organisations. It was composed of CEDEX, IEO, AFB, CEREMA, Shom, CORILA and its affiliated entities IUAV and CNR-ISMAR, MIT, IMELS, PA, CPMR, UNEP-MAP and its affiliated entity UNEP-MAP/PAP-RAC. Shom acted as coordinator. The objectives of the SIMWESTMED project were addressed through a variety of activities and desktop or case studies. They are dedicated to identifying the methodology steps, and explore the challenges and opportunities of the MSP implementation in the Western Mediterranean area, including thus related to transboundary issues (Ecosystem based approach, marine policies, Barcelona Convention, Land Sea Interactions, geographical scale of the plans, data interoperability, tools to support MSP). The project led to a multiplicity of outputs including overviews of MSP relevant information related to the countries and on more focus areas, to a number of interviews and meetings where stakeholder views were collected to feed the reasoning, and to guidelines and good practices to be shared at a national and transnational level with marine stakeholders, scientists as well as planners, administrations and authorities. In addition, SIMWESTMED permitted a lot of progression internally in the countries and regarding transboundary cooperation. It led to establish and develop new dialogues and to connect the technical or scientific actors, the stakeholders, the administrations of the countries of a same sea basin, and the administrations within the countries, including the representative of Regions. It allowed to better understand Maritime Spatial Planning mechanisms, to share knowledge and as such reached to build capacities, which is of importance as there is such a need in the Mediterranean area compared to more Northern countries. The project also permitted to address topics which have never been addressed before. The involvement of some countries in SIMWESTMED and in the EU-DG Mare "brother" projects SUPREME, SIMNORAT and SIMCelt was useful for them to develop a global vision with their neighbours through the East and West side of the Mediterranean and in the Atlantic sea basin. At the end of this exercise, it is stated the need of pursuing the work and dialogue in particular through common tools, but at this stage, the SIMWESTMED has constituted a common knowledge and background. Disclaimer: The contents and conclusions of this report, including the maps and figures, do not imply the expression of any opinion or endorsement of the participating partners concerning the legal status of any country, territory, area, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. The depiction and use of boundaries, geographic names and related data shown on maps included in this report are not warranted to be error free nor do they imply official endorsement or acceptance by any of the participating partners. This report is a working document and may rely on data from sources external to the SIMWESTMED project Consortium and, in addition to this, it may contain some information gaps. Neither the European Commission or Executive Agency for Small and Medium-sized Enterprises nor UN Environment/MAP Barcelona Convention Secretariat may be held responsible for any use that may be made of the information contained in this report.

Component: C 1.1 Initial Assessment: Developing an Overview Sub-component: C 1.1.1 Develop a basin scale analysis/initial assessment strongly MSP oriented This Report has been created thanks to the collaboration of all the Member States involved in the SIMWESTMED project that have been invited to complete the Country Fiche, a document that has led to the development of shared knowledge regarding the marine area considered in the project. Thus, the aim of this Report is to entail a collection of information across the European countries of the Western Mediterranean region and the Strait of Sicily, including Malta waters. The Initial Assessment, in fact, provides an initial overview of the area’s characteristics and this report is the harmonized output of all available information including the description of marine environment, maritime activities, key sectoral and socio-economic trends and emerging pressures, legal and transboundary issues, and governance aspects. The assessment uses existing information by organizing them in a comparable way in order to carry out a previous analysis on the main driver and issues that need to be considered for future MSP processes. The Initial Assessment is based mainly on desk-based reviews, in order to build a shared synthetic view on the Western Mediterranean region, identifying key issues (main activities and priority conservation issues) and data gaps that are synthesized in the following report., This report/document was produced as part of the SIMWESTMED Project (Grant Agreement N0. EASME/EMFF/2015/1.2.1.3/02/SI2.742101). PROJECT: Supporting Implementation of Maritime Spatial Planning in the Western Mediterranean region (SIMWESTMED) Competition for maritime space – for renewable energy equipment, aquaculture and other uses – has highlighted the need to manage our waters more coherently. Maritime spatial planning (MSP) works across borders and sectors to ensure human activities at sea take place in an efficient, safe and sustainable way. That is why the European Parliament and the Council have adopted a legislation to create a common framework for maritime spatial planning in Europe. The Directive 2014/89/EU of the European Parliament and of the Council of 23 July 2014 (said Maritime Spatial Planning Directive) establishes a framework in order to reduce conflicts between sectors and create synergies between different activities, to encourage investment – by creating predictability, transparency and clearer rules, to increase cross-border cooperation – between EU countries to develop energy grids, shipping lanes, pipelines, submarine cables and other activities, but also to develop coherent networks of protected areas, and to protect the environment – through early identification of impact and opportunities for multiple use of space. The SIMWESTMED project (Supporting Implementation of Maritime Spatial Planning in the Western Mediterranean region) is an EU/DG Mare co-funded cross-border project. It was launched on 1st of January 2017 and involves Spain, France, Italy and Malta, while these countries had just designated their Competent Authorities and transposed the Maritime Spatial Planning (MSP) Directive. SIMWESTMED aims to support the implementation of the MSP Directive in the waters of Spain, France, Italy and Malta, as well as to establish cross-border cooperation mechanisms between these Member States, to contribute to the coherence of their marine spatial plans to be established by 2021. The action ran until 31st of December 2018 and was based on a partnership of public bodies of the countries and two international organisations. It was composed of CEDEX, IEO, AFB, CEREMA, Shom, CORILA and its affiliated entities IUAV and CNR-ISMAR, MIT, IMELS, PA, CPMR, UNEP-MAP and its affiliated entity UNEP-MAP/PAP-RAC. Shom acted as coordinator. The objectives of the SIMWESTMED project were addressed through a variety of activities and desktop or case studies. They are dedicated to identifying the methodology steps, and explore the challenges and opportunities of the MSP implementation in the Western Mediterranean area, including thus related to transboundary issues (Ecosystem based approach, marine policies, Barcelona Convention, Land Sea Interactions, geographical scale of the plans, data interoperability, tools to support MSP). The project led to a multiplicity of outputs including overviews of MSP relevant information related to the countries and on more focus areas, to a number of interviews and meetings where stakeholder views were collected to feed the reasoning, and to guidelines and good practices to be shared at a national and transnational level with marine stakeholders, scientists as well as planners, administrations and authorities. In addition, SIMWESTMED permitted a lot of progression internally in the countries and regarding transboundary cooperation. It led to establish and develop new dialogues and to connect the technical or scientific actors, the stakeholders, the administrations of the countries of a same sea basin, and the administrations within the countries, including the representative of Regions. It allowed to better understand Maritime Spatial Planning mechanisms, to share knowledge and as such reached to build capacities, which is of importance as there is such a need in the Mediterranean area compared to more Northern countries. The project also permitted to address topics which have never been addressed before. The involvement of some countries in SIMWESTMED and in the EU-DG Mare "brother" projects SUPREME, SIMNORAT and SIMCelt was useful for them to develop a global vision with their neighbours through the East and West side of the Mediterranean and in the Atlantic sea basin. At the end of this exercise, it is stated the need of pursuing the work and dialogue in particular through common tools, but at this stage, the SIMWESTMED has constituted a common knowledge and background. Disclaimer: The contents and conclusions of this report, including the maps and figures, do not imply the expression of any opinion or endorsement of the participating partners concerning the legal status of any country, territory, area, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. The depiction and use of boundaries, geographic names and related data shown on maps included in this report are not warranted to be error free nor do they imply official endorsement or acceptance by any of the participating partners. This report is a working document and may rely on data from sources external to the SIMWESTMED project Consortium and, in addition to this, it may contain some information gaps. Neither the European Commission or Executive Agency for Small and Medium-sized Enterprises nor UN Environment/MAP Barcelona Convention Secretariat may be held responsible for any use that may be made of the information contained in this report.

Imposex – the superimposition of male sexual organs (penis and vas deferens) onto female Neogastropods such as Hexaplex trunculus (Linne´ , 1758) – is used world-wide as a biomarker of ecological impact of organotin based antifouling biocides (TBT and TPhT). To limit the impact of organotin pollution, since January 1, 2003, the International Maritime Organization (IMO) has enacted a global ban on the use of organotin compounds in antifouling systems. It is important to record imposex levels and organotin contamination before the implementation of the ban, in order to assess the current situation and be able, in the future, to verify the effects of the International Protocol. In this paper, recent imposex data measured in populations of Hexaplex trunculus from three different Mediterranean regions are compared: the Ligurian Sea (Italy), the Lagoon of Venice (Italy) and the western coast of Istria (Croatia). In the two former locations, a partial ban on TBT has been in force for vessels less than 25 m since 1982, while in the latter region no restrictions on organotin antifouling paints have been applied yet. Gastropod samples collected from the Venice lagoon were analysed with an acid extraction followed by Grignard derivatisation, clean up and GC-MS determination, in order to relate the levels of TBT, TPhT and their metabolites with the imposex degree detected. Biological data show that the levels of imposex were very high (VDS from 4.3 to 5) in all the sampling sites considered, particularly in the Croatian coast stations. The concentrations of organotin compounds – butyltins and phenyltins – measured in the samples from the lagoon of Venice were found to partition differently in the visceral coil and in the rest of the soft body of the analysed organisms. Imposex - the superimposition of male sexual organs (penis and vas deferens) onto female Neogastropods such as Hexaplex trunculus (Linne, 1758) - is used world-wide as a biomarker of ecological impact of organotin based antifouling biocides (TBT and TPhT). To limit the impact of organotin pollution, since January 1, 2003, the International Maritime Organization (IMO) has enacted a global ban on the use of organotin compounds in antifouling systems. It is important to record imposex levels and organotin contamination before the implementation of the ban, in order to assess the current situation and be able, in the future, to verify the effects of the International Protocol. In this paper, recent imposex data measured in populations of Hexaplex trunculus from three different Mediterranean regions are compared: the Ligurian Sea (Italy), the Lagoon of Venice (Italy) and the western coast of Istria (Croatia). In the two former locations, a partial ban on TBT has been in force for vessels less than 25 m since 1982, while in the latter region no restrictions on organotin antifouling paints have been applied yet. Gastropod samples collected from the Venice lagoon were analysed with an acid extraction followed by Grignard derivatisation, clean up and GC-MS determination, in order to relate the levels of TBT, TPhT and their metabolites with the imposex degree detected. Biological data show that the levels of imposex were very high (VDS from 4.3 to 5) in all the sampling sites considered, particularly in the Croatian coast stations. The concentrations of organotin compounds - butyltins and phenyltins - measured in the samples from the lagoon of Venice were found to partition differently in the visceral coil and in the rest of the soft body of the analysed organisms.

In this study, we screened eight terpenes isolated from the organic extract of Sphaerococcus coronopifolius for their antifouling activity in order to find possible new sources of non-toxic or less toxic bioactive antifoulants. The anti-settlement activity (EC 50) and the degree of toxicity (LC 50) of S. coronopifolius metabolites was evaluated using larvae of the cirriped crustacean Amphibalanus (Balanus) amphitrite (cyprids and nauplii) as model organism. For five of eight tested metabolites EC 50 was lower than 5 mg/L. The most promising results were observed for bromosphaerol (3), which expressed an EC 50 value of 0.23 mg/L, in combination with low toxicity levels (LC 50 > 100 mg/L). The therapeutic ratio-an index used to estimate whether settlement inhibition is due to toxicity or other mechanisms-is also calculated and discussed. © 2010 Springer Science+Business Media, LLC.

The discharge of industrial and municipal wastewaters containing substances potentially harmful to aquatic ecosystems and human health represents one of the major problems connected to water management. The efforts towards this threat reduction must go hand in hand with the development of monitoring systems able to provide, on-line and in real time, an early-warning information about the presence of pollutants in the water. Starting from these considerations, the electrochemical activity of natural water biofilms, largely studied in the last 20 years, has been exploited to develop two different kinds of biosensors for industrial and environmental applications. In the first case, since large amounts of disinfectants and other chemical substances are employed in the water lines of industrial plants as a countermeasure against biofilm, aim of this research was to realize a sensor for bacterial growth monitoring, to optimize the above mentioned biofilm cleaning treatments. This optimization, indeed, entails a reduction of both costs and of biocide treatments environmental impact. In the second case, considering that the biofilm is a micro-ecosystem, the changes induced in its electrochemical activity by addition in water of toxic substances were studied, to realize a biosensor for water pollution, based on this phenomenon. Concerning the industrial application, it emerged that, adjusting biocide treatments basing on the real biofilm growth monitored by this sensor prototype, it was possible to drastically reduce the use of chemicals within industrial water lines. The biosensor for water pollution provided interesting results too; when different toxic substances were added to the water in which the biofilm was growing, the bacterial electrochemical signal was inhibited, and this inhibition was proportional to the concentration of the added toxic substance.

In recent years massive efforts have been directed to identify ecologically relevant invertebrate toxicity testing models. The crustacean Amphibalanus amphitrite represent a valid model for ecotoxicology studies, as it is widely distributed, ecologically relevant and its larvae can be easily obtained and reared in laboratory. The UNICHIM standardization process of the acute toxicity assay with larvae (II stage nauplii) of A. amphitrite has recently been concluded. Aim of this work is to show the possibility of obtaining three distinct end-points (acute and sub-lethal), characterized by different sensitivity levels, by the use of the same model organism and the preparation of just one bioassay. Marine sediments (elutriates and whole sediment) were used as an example of environmental sample.

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Nell'ambito di un recente programma di ricerca finanziato dalla Regione Veneto e coordinato da ARPA Veneto-Osservatorio Alto Adriatico (93-T406-REL-G009, Intervento n.72-Campo Sperimentale in Mare), due nuovi test tossicologici di tipo comportamentale (il primo basato sulla valutazione delle caratteristiche del riaffossamento di molluschi bentonici della specieTapes philippinarum, il secondo sull'alterazione del nuoto larvale del crostaceo Balanus amphitrite) sono stati applicati sperimentalmente su 9 campioni di sedimento marino-costiero prelevati lungo la costa veneta. I risultati ottenuti, confrontati con le risposte di alcuni test tossicologici indicati dalla normativa per il monitoraggio marino-costiero (Microtox ®e inibizione della crescita algale di Dunaliella tertiolecta) e con la valutazione analitica del contenuto di numerosi contaminanti nei sedimenti, dimostrano come una batteria di saggi tossicologici che comprenda anche test innovativi sia lo strumento che meglio riesce ad evidenziare e discriminare la tossicità dei sedimenti