Cytogenetic characterisation of the razor shells "Ensis directus" (Conrad, 1843) and "E. minor" (Chenu, 1843) (Mollusca: Bivalvia)

[Abstract] The European razor shell Ensis minor (Chenu 1843) and the American E. directus (Conrad 1843) have a diploid chromosome number of 38 and remarkable differences in their karyotypes: E. minor has four metacentric, one metacentric–submetacentric, five submetacentric, one subtelocentric and eight telocentric chromosome pairs, whereas E. directus has three metacentric, two metacentric–submetacentric, six submetacentric, six subtelocentric and two telocentric pairs. Fluorescent in situ hybridisation (FISH) using a major ribosomal DNA probe located the major ribosomal genes on one submetacentric chromosome pair in both species; FISH with a 5S ribosomal DNA (5S rDNA) probe rendered one chromosomal (weak) signal for E. minor and no signal for E. directus, supporting a more dispersed organisation of 5S rDNA compared to the major ribosomal genes. The vertebrate telomeric sequence (TTAGGG)n was located on both ends of each chromosome, and no interstitial signals were detected. In this work, a comparative karyological analysis was also performed between the four Ensis species analysed revealing that the three European species studied so far, namely E. minor, E. siliqua (Linné 1758) and E. magnus Schumacher 1817 show more similarities among them than compared to the American species E. directus. In addition, clear karyotype differences were found between the morphologically similar species E. minor and E. siliqua.Ministerio de Educación y Ciencia; CTM2007-28919-E/MA

An efficient method to find potentially universal population genetic markers, applied to metazoans

<p>Abstract</p> <p>Background</p> <p>Despite the impressive growth of sequence databases, the limited availability of nuclear markers that are sufficiently polymorphic for population genetics and phylogeography and applicable across various phyla restricts many potential studies, particularly in non-model organisms. Numerous introns have invariant positions among kingdoms, providing a potential source for such markers. Unfortunately, most of the few known EPIC (Exon Primed Intron Crossing) loci are restricted to vertebrates or belong to multigenic families.</p> <p>Results</p> <p>In order to develop markers with broad applicability, we designed a bioinformatic approach aimed at avoiding multigenic families while identifying intron positions conserved across metazoan phyla. We developed a program facilitating the identification of EPIC loci which allowed slight variation in intron position. From the <it>Homolens </it>databases we selected 29 gene families which contained 52 promising introns for which we designed 93 primer pairs. PCR tests were performed on several ascidians, echinoderms, bivalves and cnidarians. On average, 24 different introns per genus were amplified in bilaterians. Remarkably, five of the introns successfully amplified in all of the metazoan genera tested (a dozen genera, including cnidarians). The influence of several factors on amplification success was investigated. Success rate was not related to the phylogenetic relatedness of a taxon to the groups that most influenced primer design, showing that these EPIC markers are extremely conserved in animals.</p> <p>Conclusions</p> <p>Our new method now makes it possible to (i) rapidly isolate a set of EPIC markers for any phylum, even outside the animal kingdom, and thus, (ii) compare genetic diversity at potentially homologous polymorphic loci between divergent taxa.</p

Histoire évolutive, structures génétique, morphologique et écologique comparées dans un complexe d'espèces jumelles : Echinocardium cordatum (Echinoidea, Irregularia)

Echinocardium cordatum (Pennant 1777) oursin irrégulier abondant des zones côtières tempérées a longtemps été considéré comme une espèce cosmopolite dont la vaste aire de distribution était la conséquence directe des capacités de dispersion de sa larve planctotrophe. L’étude couplée des caractéristiques génétiques [génomes mitochondrial et nucléaire (introns+microsatellites)], morphologiques (étude basée sur 20 indices morphométriques) et écologiques (distribution géographique à petite ou grande échelle, et cycle de maturation gonadique) a révélé la présence d’un complexe d’espèces jumelles dont la différenciation génétique est accompagnée d’une différenciation morphologique statistique ainsi que de différenciations écologiques plus ou moins fines. Ces espèces occupent des aires de distribution limitées (clade A : Atlantique, clade SP : Pacifique Sud, clade NP : Pacifique Nord, clade B2 : Méditerranée, et clade B1 : Méditerranée et côtes atlantiques de l’Ibérie). D’après la reconstruction de l’histoire évolutive de ce complexe, à partir des données paléontologiques et moléculaires, ces espèces auraient divergé il y a 3 (B1-B2) à 10 (A-reste) millions d’années sous l’effet de perturbations géologiques et paléoclimatiques (fermeture de la Téthys, crise messinienne de salinité et glaciations Plio-Pléistocène). Le polymorphisme morphologique et moléculaire apparaît réduit chez B1 suggérant un effectif efficace historique de cette espèce réduit. L’analyse des flux géniques contemporains révèle que les clades A et B1 échangent toujours des gènes, alors que les clades B1 et B2, ont mis en place un isolement reproducteur efficace empêchant l’hybridation. Par ailleurs, les capacités de dispersion des espèces de ce complexe sont importantes (plus de 3000 km), mais moindres comparées à d’autres espèces du genre, notamment E. mediterraneum, qui bien qu’ayant subi les mêmes évènements géologiques n’a pas formé d’espèce depuis son apparition il y aurait 28 millions d’années. D’un point de vue évolutif, les taxons à forte capacité de dispersion présenteraient des tailles efficaces de populations importantes, ainsi qu’une aire de répartition étendue et peu de différentiation génétique entre localités ; autant de caractéristiques qui devraient ralentir la vitesse de spéciation dans ces taxons. Si cette hypothèse semble se vérifier chez E. mediterraneum, il n’en est pas de même chez E. cordatum qui malgré des effectifs efficaces apparemment importants et une différenciation des populations à l’échelle régionale faible, présente une dynamique se spéciation plus rapide. Il faut envisager que d’autres caractéristiques soient à l’origine de cette différence de dynamique de spéciation, et la comparaison des exigences écologiques des deux taxons ainsi que l’isolement de la molécule responsable de la réaction acrosomique, la bindine, pourraient apporter des éléments de réponse aux nouvelles questions soulevées.Echinocardium cordatum (Pennant 1777) an abundant irregular sea urchin from the coastal temperate zones has long been considered as a cosmopolitan species which wide distribution area was the direct consequence of its planktotrophic larvae high dispersal abilities. A combined study of the genetic [mitochondrial and nuclear genomes (introns+microsatellites)], morphologic (based on 20 morphometric indices) and ecologic (geographic distribution at fine or large scale, and gonad maturation cycle) characteristics reveals that this taxon is a complex of cryptic species for which genetic differentiations concurred with morphological and ecological ones. The different species each occupy a limited geographic areas (clade A : Atlantic, clade SP : South Pacific, clade NP : North Pacific, clade B2 : Mediterranean sea, et clade B1 : Mediterranean sea and Atlantic coasts of Iberia). According to the complex species evolutionary history reconstruction, based on fossils and molecular data, the different species diverged between 3 (B1-B2) and 10 (A-rest) million years ago, driven by geologic and paleoclimatic perturbations (Tethys closure, messinian salinity crisis, Plio-Pleistocene glaciations). Molecular and morphologic polymorphisms appear reduced in B1, suggesting a reduced historical effective size. The contemporaneous genetic flux analysis reveals that clades A and B1 exchange genes whereas clades B1 and B2 developed an efficient reproductive isolation preventing hybridization. Though dispersal abilities of the complex species are high (more than 3000 km), they appear to be smaller than those of other species of the same genera, particularly E. mediterraneum which undergone the same geological perturbations without splitting into several species since its appearance some 28 million years ago. From an evolutionary point of view, taxa with high dispersal abilities should exhibit important population effective sizes, wide distribution areas and weak genetic differentiation between localities, properties that should slow species formation within these taxa. If this hypothesis seems verified in E. mediterraneum, it is not the case in E. cordatum for which the apparent high effective size and weak regional structure contrast with the fast speciation dynamics. It seems that other characteristics might be responsible for the speciation dynamic differences, and the comparison of the two taxa ecological requirements, as well as the isolation of the gene coding for the protein responsible of the sperm specific attachment, the bindin, should bring elements to answer these questions

Histoire évolutive, structures génétique, morphologique et écologique comparées dans un complexe d'espèces jumelles (Echinocardium cordatum (Echinoidea, Irregularia))

Echinocardium cordatum (Pennant 1777) oursin irrégulier abondant des zones côtières tempérées a longtemps été considéré comme une espèce cosmopolite dont la vaste aire de distribution était la conséquence directe des capacités de dispersion de sa larve planctotrophe. L étude couplée des caractéristiques génétiques [génomes mitochondrial et nucléaire (introns+microsatellites)], morphologiques (étude basée sur 20 indices morphométriques) et écologiques (distribution géographique à petite ou grande échelle, et cycle de maturation gonadique) a révélé la présence d un complexe d espèces jumelles dont la différenciation génétique est accompagnée d une différenciation morphologique statistique ainsi que de différenciations écologiques plus ou moins fines. Ces espèces occupent des aires de distribution limitées (clade A : Atlantique, clade SP : Pacifique Sud, clade NP : Pacifique Nord, clade B2 : Méditerranée, et clade B1 : Méditerranée et côtes atlantiques de l Ibérie). D après la reconstruction de l histoire évolutive de ce complexe, à partir des données paléontologiques et moléculaires, ces espèces auraient divergé il y a 3 (B1-B2) à 10 (A-reste) millions d années sous l effet de perturbations géologiques et paléoclimatiques (fermeture de la Téthys, crise messinienne de salinité et glaciations Plio-Pléistocène). Le polymorphisme morphologique et moléculaire apparaît réduit chez B1 suggérant un effectif efficace historique de cette espèce réduit. L analyse des flux géniques contemporains révèle que les clades A et B1 échangent toujours des gènes, alors que les clades B1 et B2, ont mis en place un isolement reproducteur efficace empêchant l hybridation. Par ailleurs, les capacités de dispersion des espèces de ce complexe sont importantes (plus de 3000 km), mais moindres comparées à d autres espèces du genre, notamment E. mediterraneum, qui bien qu ayant subi les mêmes évènements géologiques n a pas formé d espèce depuis son apparition il y aurait 28 millions d années. D un point de vue évolutif, les taxons à forte capacité de dispersion présenteraient des tailles efficaces de populations importantes, ainsi qu une aire de répartition étendue et peu de différentiation génétique entre localités ; autant de caractéristiques qui devraient ralentir la vitesse de spéciation dans ces taxons. Si cette hypothèse semble se vérifier chez E. mediterraneum, il n en est pas de même chez E. cordatum qui malgré des effectifs efficaces apparemment importants et une différenciation des populations à l échelle régionale faible, présente une dynamique se spéciation plus rapide. Il faut envisager que d autres caractéristiques soient à l origine de cette différence de dynamique de spéciation, et la comparaison des exigences écologiques des deux taxons ainsi que l isolement de la molécule responsable de la réaction acrosomique, la bindine, pourraient apporter des éléments de réponse aux nouvelles questions soulevées.Echinocardium cordatum (Pennant 1777) an abundant irregular sea urchin from the coastal temperate zones has long been considered as a cosmopolitan species which wide distribution area was the direct consequence of its planktotrophic larvae high dispersal abilities. A combined study of the genetic [mitochondrial and nuclear genomes (introns+microsatellites)], morphologic (based on 20 morphometric indices) and ecologic (geographic distribution at fine or large scale, and gonad maturation cycle) characteristics reveals that this taxon is a complex of cryptic species for which genetic differentiations concurred with morphological and ecological ones. The different species each occupy a limited geographic areas (clade A : Atlantic, clade SP : South Pacific, clade NP : North Pacific, clade B2 : Mediterranean sea, et clade B1 : Mediterranean sea and Atlantic coasts of Iberia). According to the complex species evolutionary history reconstruction, based on fossils and molecular data, the different species diverged between 3 (B1-B2) and 10 (A-rest) million years ago, driven by geologic and paleoclimatic perturbations (Tethys closure, messinian salinity crisis, Plio-Pleistocene glaciations). Molecular and morphologic polymorphisms appear reduced in B1, suggesting a reduced historical effective size. The contemporaneous genetic flux analysis reveals that clades A and B1 exchange genes whereas clades B1 and B2 developed an efficient reproductive isolation preventing hybridization. Though dispersal abilities of the complex species are high (more than 3000 km), they appear to be smaller than those of other species of the same genera, particularly E. mediterraneum which undergone the same geological perturbations without splitting into several species since its appearance some 28 million years ago. From an evolutionary point of view, taxa with high dispersal abilities should exhibit important population effective sizes, wide distribution areas and weak genetic differentiation between localities, properties that should slow species formation within these taxa. If this hypothesis seems verified in E. mediterraneum, it is not the case in E. cordatum for which the apparent high effective size and weak regional structure contrast with the fast speciation dynamics. It seems that other characteristics might be responsible for the speciation dynamic differences, and the comparison of the two taxa ecological requirements, as well as the isolation of the gene coding for the protein responsible of the sperm specific attachment, the bindin, should bring elements to answer these questions.AIX-MARSEILLE2-Bib.electronique (130559901) / SudocSudocFranceF

Does Hybridization Increase Evolutionary Rate? Data from the 28S-rDNA D8 Domain in Echinoderms

The divergent domain D8 of the large ribosomal RNA is very variable and extended in vertebrates compared to other eukaryotes. We provide data from 31 species of echinoderms and present the first comparative analysis of the D8 in nonvertebrate deuterostomes. In addition, we obtained 16S mitochondrial DNA sequences for the sea urchin taxa and analyzed single-strand conformation polymorphism (SSCP) of D8 in several populations within the species complex Echinocardium cordatum. A common secondary structure supported by compensatory substitutions and indels is inferred for echinoderms. Variation mostly arises at the tip of the longest stem (D8a), and the most variable taxa also display the longest and most stable D8. The most stable variants are the only ones displaying bulges in the terminal part of the stem, suggesting that selection, rather than maximizing stability of the D8 secondary structure, maintains it in a given range. Striking variation in D8 evolutionary rates was evidenced among sea urchins, by comparison with both 16S mitochondrial DNA and paleontological data. In Echinocardium cordatum and Strongylocentrotus pallidus and S. droebachiensis, belonging to very distant genera, the increase in D8 evolutionary rate is extreme. Their highly stable D8 secondary structures rule out the possibility of pseudogenes. Thes

Differential reproductive timing in Echinocardium spp.: The first Mediterranean survey allows interoceanic and interspecific comparisons

International audienceEchinocardium cordatum had long been considered as cosmopolitan, but molecular data revealed it is a complex of cryptic species, with two non-hybridizing species (B1 & B2) in the Mediterranean Sea living in syntopy with Echinocardium mediterraneum. Histological analyses of the gonads from a 17-month sampling period revealed a statistically significant time lag between the Maturity Indices of E. cordatum and E. mediterraneum. The main environmental stimulus may be different for the two nominal species, possibly seawater temperature for E. cordatum and chlorophyll a concentration for E. mediterraneum. Within the E. cordatum complex, spawning timing and synchrony are different according to major geographic areas (Atlantic/Pacific/Mediterranean) and/or the corresponding genetic subdivision [A/P/(B1 & B2)]. In contrast, the effects of temperature on the reproductive cycle seem rather to mirror the genetic lineages than environmental similarities of the different localities. Between the sister species (B1 & B2) no differences could be detected, maybe due to small sample sizes. (c) 2010 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved

Morphological and genetic analyses reveal a cryptic species complex in the echinoid Echinocardium cordatum and rule out a stabilizing selection explanation.

International audiencePreliminary analyses revealed the presence of at least five mitochondrial clades within the widespread sea urchin Echinocardium cordatum (Spatangoida). In this study, we analyzed the genetic (two mitochondrial and two nuclear sequence loci) and morphological characteristics (20 indices) from worldwide samples of this taxon to establish the species limits, morphological diversity and differentiation. Co-occurring spatangoid species were also analyzed with mitochondrial DNA. The nuclear sequences confirm that mitochondrial lineages correspond to true genetic entities and reveal that two clades (named A and B1) hybridize in their sympatry area, although a more closely related pair of clades (B1 and B2), whose distributions widely overlap, does not display hybridization. The morphology of all E. cordatum clade pairs was significantly differentiated, but no morphological diagnostic character was evidenced. By contrast, other spatangoid species pairs that diverged more recently than the E. cordatum clades display clear diagnostic characters. Morphological diversity thus appears responsible for the absence of diagnostic characters, ruling out stabilizing selection, a classical explanation for cryptic species. Alternative classical explanations are (i) environmental plasticity or (ii) a high diversity of genes determining morphology, maintained by varying environmental conditions. We suggest a new hypothesis that the observed morphological diversity is selectively neutral and reflects high effective population sizes in the E. cordatum complex. It is supported by the higher abundance of this taxon compared with other taxa, a trend for the genetic and morphological diversity to be correlated in Europe, and the higher genetic and morphological diversities found in clades of E cordatum (except B1) than in other spatangoid samples in Europe. However, the Pacific clades do not confirm these trends

Biodiversity and Food Security: From Trade-offs to Synergies

International audienceBiodiversity and food security are connected in many ways. Across scales from genes to species, landscapes, and biomes, biodiversity is an important resource for humanity. It is the key for a broad range of services provided by ecosystems. Biodiversity helps regulate the nutrient cycle and water (e.g., floods) and mitigates impacts of climate change. Biodiversity is also of direct importance for human well-being and for cultural and other values including recreation. The provisioning of clean water and diverse food supply makes it vital for all people. Biodiversity at all levels, including the diversity of genes, species, and ecosystems, is lost at alarming rates. Critical factors for these trends are habitat destruction, global warming, and the uncontrolled spread of alien species. Pollution, nitrogen deposition, and shifts in precipitation further affect biodiversity. Food security faces significant challenges due to population growth, poverty, globalization, climate change, and other factors. Supplying healthy food to all citizens is crucial for global development—to reach it, not only food production but also equitable access to food for all people must be improved substantially. Biodiversity loss and global food security are hence two major challenges of our time. Linking these two areas from a research perspective and seeking synergies between them are likely to generate multiple benefits for social, ecological, and economic development

CIGESMED : Coralligenous based Indicators to evaluate and monitor the "Good Environmental Status" of the Mediterranean coastal waters

Final reportCoralligenous is a hard-bottom mainly biogenic habitat, produced by the agglomeration of calcareous encrusting algae growing in dim-light conditions. It is characterized by high structural complexity and spatial heterogeneity, thus supporting rich biodiversity and a variety of sessile assemblages, shaping a typical and one of the most important habitats of the Mediterranean Sea. It produces goods (e.g. food, raw material) and services in several domains (e.g. CO2 sequestration, aesthetics and education). Pollution, smothering and abrasion from a variety of human activities may cause its degradation at a broad scale, whilst fishing and collection of organisms mainly affect target species. Its high aesthetic value may also induce frequentation by SCUBA divers, an additional cause of degradation. Coralligenous is also susceptible to invasive alien species. This habitat, which is of great ecological, socio-economic and cultural importance, is also under the pressures linked to global warming.CIGESMED’s GOAL was to understand links between natural and anthropogenic pressures and coralligenous habitats as well as the effects on their functioning to define the Good Environmental Status (GES) of the coastal Mediterranean Sea and propose solutions for maintaining good environmental conditions.Coralligenous specific indices have been constructed and tested by scientists, marine natural parks and reserves managers, also through the implementation of a “citizen science” pilot network. The use of the newest data mining techniques and the development of visualization tools to sort, organize and illustrate very large heterogeneous sets of data constitute an original but complex approach. It permitted to mobilize, visualize and share large data collections, and to manage knowledge to study these habitats.The OUTCOME consists of: i) experimentation and results of new methods to build survey at large scales (testing operating process and materials during dive, photo analyses, population genetics, phylogenetic and metabarcoding approaches) ii) tools to diffuse new methods (website, services, training and field tools for scientists and citizen science, publications) and avoid indices misun-derstanding, iii) tools, methods and prototypes to provide datamining usable for an integrative assessment of the GES within the Framework of the Marine Strategy Directive (for this part, CIGESMED members initiated a new consortium using CIGESMED metadata and dataset to build graph representation, mine graphs and provide tools for environmental decision making). All the outcomes are freely accessible online on websites with open access, open source and open data.The overall achievement was to to bring together researchers (in ecology, economics, sociology, law, etc.) and managers in order to (i) identify the needs and to better address them, (ii) to determine interdisciplinary areas of research concerning the development and management of the coralligenous that could be the subject of a new [multidisciplinary / European / Mediterranean] research project.CIGESMED gathered scientists from France, Greece and Turkey, making it possible to assess the coralligenous habitat in a number of sites in both the northwestern Mediterranean basin and the Aegean-Levantine, under a common approach. Members of ten highly experienced marine ecology laboratories were involved.A total of 10 stations in France (in the Gulf of Lions), Greece (in the Ionian and the Aegean Seas) and Turkey (in the Aegean and the Levantine Seas) were selected to study coralligenous assemblages across the Mediterranean Sea. Analyses of photoquadrats (50x50 cm) and in situ visual observations revealed 313 species, belonging to 15 higher taxa. A total of 204 species were found in Turkey, 192 species in France and 109 species in Greece. Only fifty species were common in all sites. The abundance of the taxonomic groups in coralligenous habitats vary among sites and countries. The multivariate analysis revealed five main assemblages across the Mediterra-nean Sea. A number of important threats were withnessed to have an important impact on coralligenous, with the settlement of invasive alien species (e.g. Caulerpa cylindracea, Womersleyella setacea), sedimentation and factors causing algal bleaching being the most important ones.A new method and index were applied and tested in France (60 stations in Gulf of Lions and Provence) and Greece (4 stations in the Gulf of Corinth) to evaluate the health condition of coralligenous assemblages. This method (INDEX-COR), based on images analyses (60x40cm) and in situ observations, takes into account three metrics giving different levels of information: (i) the ratio "Sensitive-Tolerante Species", (ii) the observable taxonomic richness of the assemblages and (iii) the structural complexity. The global index combining these metrics was tested according to a global index of pressure. Reference conditions were defined in France to propose an interpretation grid to evaluate the status of coralligenous assemblages. This grid applied in Greece demonstrated the necessity to collecte multiple and complete dataset in order to define the reference conditions for the different Mediterranean sectors (e.g. Ionian Sea, Egean Sea and Cretan Sea). Finally, additional indices (CAI, COARSE and ESCA) were tested on the datasets obtained in France and Greece. The comparison of the results is still in progress. The first ones show the advantages and the limits of each index. They underline the need to achieve precisions on (i) their degree of sensitivity in the evaluation of the coralligenous assemblages and (ii) the impact of the different images analysis techniques.Although the global set of samples is still under analysis, the metabarcoding pilot study already gave very promising results for the assessment of coralligenous community species composition: many more species were identified than by eye, predicting a higher resolution than traditional approaches for monitoring and comparing coralligenous assemblages.A dedicated Citizen Science initiative was designed and launched in the course of the project, aiming to engage enthusiast divers in the study and monitoring of coralligenous assemblages through the gathering of basic information regarding spatial occurrence, assemblage structure and associated pressures and threats. The implementation platform comprises a data collection protocol and a multilingual website which serve both educational and data submission purposes. Online and paper educational documentation, as well as observation protocol guidelines are essential tools developed to train volunteer divers. Underwater slates based on the princi-ples of rapid visual assessment have also been developed and distributed to all participants for data collection. Geo-referenced data reporting focuses on: (a) basic topographic and abiotic features for the preliminary description of each site, and the creation of data series for sites receiving multiple visits; (b) presence and relative abundance of typical conspicuous species, as well as, (c) existence of pressures and imminent threats, for the characterization and assessment of coralligenous assemblages. More than 100 observation sets from across the Mediterranean have been registered to date by approximately 30 divers while 75 members have registered to the website, which remain active after the conclusion of the project.Metadata and data produced by the CIGESMED project have a high potential for use by several stakeholders involved in environmental management. Mapping this information needed to share common definitions on coralligenous components and allows starting building a micro thesaurus. The methodology is now developed and the first part of the thesaurus is online. A new consortium called IndexMed whose task is to index Mediterranean biodiversity data, makes it possible to build graphs in order to analyse the CIGESMED data and develop new solutions for coralligenous data mining.The outreach activity of the project included participation of the Consortium to an impressive number of Conferences, Symposia and Workshops, which made it possible to achieve not only the communication of the main objectives of the project but also the development of links with other projects, targeting Marine Biodiversity [e.g. EMBOS (The European Marine Biodiversity Observa-tory), LifeWatch (ESFRI Research Infrastruc-ture), DEVOTES (DEVelopment Of innovative Tools for understanding marine biodiversity and assessing good Environmental Status), VECTORS (VECTORS of Change in European Marine Ecosystems and their Environmental and Socio-Economic Impacts) and EU BON (Building the European Biodiversity Observation Net-work)]. A flyer was developed to provide the basic information on the project. The project was also advertised during much larger events, in the premises of the participating institutes, such as open days (e.g. exhibitions for the Climate change, TEDx events). The target audience for the outreach activity consisted of researchers and scientists, students, educators, environmental managers, policy makers and stakeholders from all the economic sectors including industry. The means which were used were: (a) the project website, which until now shows a high number of visits recorded for a specialized one: 250,000 hits; (b) the production of 353 articles for the scientific audience and for the society at large. Five websites have been created from which information on the project is broadcasted.The methods and datasets produced by CIGESMED are disseminated to the STIC community, as free tools for studies to be used for any type of data sciences (data mining, data representation ...), particularly through the means provided by IndexMed and through data qualification processes (which will need to be continuously improved to keep them relevant). This reusability of the data will be improved in particular by the production of data papers and future animations planned within the framework of the IndexMed consortiumThe Steering Committee consisting of all the WP leaders and the coordinator was responsible for all practical decision making, strategic planning and implementation.A Committee of External Advisors met at an annual basis, and aimed at providing advice on all aspects of the execution of the project to ensure CIGESMED to meet its objectives