Insights on Pinna nobilis population genetic structure in the Aegean and Ionian Sea.

The fan mussel Pinna nobilis Linnaeus, 1758 is an endemic species of the Mediterranean Sea, protected by international agreements. It is one of the largest bivalves in the world, playing an important role in the benthic communities; yet it has been recently characterized as Critically Endangered by the IUCN, due to mass mortality events. In this context, the assessment of the genetic variation of the remaining P. nobilis populations and the evaluation of connectivity among them are crucial elements for the conservation of the species. For this purpose, samples were collected from six regions of the Eastern Mediterranean Sea; the Islands of Karpathos, Lesvos and Crete; the Chalkidiki and Attica Peninsulas; and the Amvrakikos Gulf. Sampling was performed either by collecting tissue from the individuals or by using a non-invasive method, i.e., by scraping the inside of their shells aiming to collect their mucus and thus avoid stress induction to them. Conventional molecular techniques with the use of the COI and 16S rRNA mitochondrial markers were selected for the depiction of the intra-population genetic variability. The analyses included 104 samples from the present study and publicly available sequences of individuals across the whole Mediterranean Sea. The results of this work (a) suggest the use of eDNA as an efficient sampling method for protected bivalves and (b) shed light to the genetic structure of P. nobilis population in the Eastern Mediterranean; this latter knowledge might prove to be fundamental for the species conservation and hence the ecosystem resilience. The haplotype analyses reinforced the evidence that there is a certain degree of connectivity among the distinct regions of the Mediterranean; yet there is evidence of population distinction within the basin, namely between the Western and the Eastern basins. The combination of both genetic markers in the same analysis along with the inclusion of a large number of individuals produced more robust results, revealing a group of haplotypes being present only in the Eastern Mediterranean and providing insights for the species' most suitable conservation management.

Micro-CT image gallery visually presenting the effects of ocean warming and acidification on marine gastropod shells.

BACKGROUND: Digitisation of specimens (e.g. zoological, botanical) can provide access to advanced morphological and anatomical information and promote new research opportunities. The micro-CT technology may support the development of "virtual museums" or "virtual laboratories" where digital 3D imaging data are shared widely and freely. There is currently a lack of universal standards concerning the publication and curation of micro-CT datasets. NEW INFORMATION: The aim of the current project was to create a virtual gallery with micro-CT scans of individuals of the marine gastropod Hexaplextrunculus, which were maintained under a combination of increased temperature and low pH conditions, thus simulating future climate change scenarios. The 3D volume-rendering models created were used to visualise the structure properties of the gastropods shells. Finally, the 3D analysis performed on the micro-CT scans was used to investigate potential changes in the shell properties of the gastropods. The derived micro-CT 3D images were annotated with detailed metadata and can be interactively displayed and manipulated using online tools through the micro-CT virtual laboratory, which was developed under the LifeWatchGreece Research Infrastructure for the dissemination of virtual image galleries collection supporting the principles of FAIR data.

Micro-CT protocols for scanning and 3D analysis of Hexaplextrunculus during its different life stages.

Micro-computed tomography (micro-CT) is a high-resolution 3D-imaging technique which is now increasingly applied in biological studies focusing on taxonomy and functional morphology. The creation of virtual representations of specimens can increase availability of otherwise underexploited and inaccessible samples. The 3D model dataset can be also further processed through volume rendering and morphometric analysis. The success of micro-CT as a visualisation technique depends on several methodological manipulations, including the use of contrast enhancing staining agents, filters, scanning mediums, containers, exposure time and frame averaging. The aim of this study was to standardise a series of micro-CT scanning and 3D analysis protocols for a marine gastropod species, Hexaplextrunculus. The analytical protocols have followed all the developmental stages of this gastropod, from egg capsules and embryos to juveniles and adults.

Micro-CT for Biological and Biomedical Studies: A Comparison of Imaging Techniques.

Several imaging techniques are used in biological and biomedical studies. Micro-computed tomography (micro-CT) is a non-destructive imaging technique that allows the rapid digitisation of internal and external structures of a sample in three dimensions and with great resolution. In this review, the strengths and weaknesses of some common imaging techniques applied in biological and biomedical fields, such as optical microscopy, confocal laser scanning microscopy, and scanning electron microscopy, are presented and compared with the micro-CT technique through five use cases. Finally, the ability of micro-CT to create non-destructively 3D anatomical and morphological data in sub-micron resolution and the necessity to develop complementary methods with other imaging techniques, in order to overcome limitations caused by each technique, is emphasised.

Benthic habitat mapping of Plazh Gradina - Zlatna ribka (Black Sea) and Karpathos and Saria Islands (Mediterranean Sea).

BACKGROUND: Habitat mapping is nеcessary for the efficient conservation and protection of marine ecosystems. In addition, it is a requirement for EU Member States as stated in the European Union (EU) Habitats Directive (92/43/EEC), as well as necessary for the achievement and maintenance of 'good environmental status (GES)' of benthic marine habitats in the framework of the EU Marine Strategy Framework Directive (2008/56/EC). NEW INFORMATION: This study provides baseline information on the marine benthic habitats of Sozopol Bay (Black Sea) and Karpathos and Saria Islands (Mediterranean Sea). These two Natura 2000 sites were selected as study sites of the RECONNECT project, which aimed at creating a transnational cooperative network to confront the environmental threats of ecosystems with a high natural and cultural interest, by the establishment of common practices and a joint regional strategy. The specific objective was to map the marine habitats using a defined a priori classification (EUNIS), with the ultimate purpose of supporting government marine spatial planning, management and decision-making processes through the development of a Decision Support System.

Benthic communities in three Mediterranean touristic ports: MAPMED project.

Background: Mediterranean ports are sources of significant economic activity and at the same time they act as recipients of considerable anthropogenic disturbance and pollution. Polluted and low-in-oxygen sediments can negatively impact benthic biodiversity and favour recruitment of opportunistic or invasive species. Macrobenthic communities are an important component of the port biota and can be used as environmental quality indicators. However, a baseline database for benthic biodiversity in Mediterranean ports has not yet been widely established. New information: Macrobenthic assemblages were recorded in three Mediterranean touristic ports under the framework of the ENPI CBC MED project MAPMED (MAnagement of Port Areas in the MEDiterranean Sea Basin). Samples were collected from Cagliari (Sardinia, Italy), Heraklion (Crete, Greece) and El Kantaoui (Tunisia) ports during February, May and September 2012. The sampling stations were selected according to the different sectors within each port (i.e. leisure, fishing, passenger/cargo vessels and shipyard). A total of 277 taxa belonging to 12 phyla were found, of which the 96 taxa were present in all three ports. El Kantaoui port hosted the highest number of macrobenthic taxa. Mollusca were the most abundant group (34%) in all ports. The highest percentage of opportunistic taxa per station was found before the touristic period in the shipyard of Heraklion port (89.3%).

Ecological status assessment based on benthic macrofauna of three Mediterranean ports: Comparisons across seasons, activities and regions.

The present study aimed to compare anthropogenic impacts in three Mediterranean ports (Cagliari-Italy, Heraklion-Greece, El-Kantatoui-Tunisia) employing benthic macrofaunal indices, used in the context of the European Water Framework Directive. Sampling stations were selected within ports according to sector usage categorization and sampled over three seasons. Benthic indices calculated include M-AMBI, BENTIX, BQI and BQI_Family. Comparisons were made between the indices values and ecological status (ES) of each station within and between ports. Overall, few statistically significant differences were observed across different seasons or different stations with the same usage within or across ports. The ES of sampling stations in the leisure/fishing and passenger/cargo ships sectors was mostly "good" or "moderate", while the shipyard sector had "poor" ES. The results suggest that the indices used were suitable for assessing the ES of Mediterranean ports, led to comparable results even across different countries and contribute to the adaptation of specific port monitoring guidelines.

Spatio-temporal benthic biodiversity patterns and pollution pressure in three Mediterranean touristic ports.

The Mediterranean Sea is one of the busiest areas worldwide in terms of maritime activity, facing considerable anthropogenic disturbance, such as pollution by hydrocarbons and heavy metals. The present study has evaluated the environmental and benthic biodiversity characteristics of three touristic ports, Cagliari (Sardinia, Italy), Heraklion (Crete, Greece) and El Kantaoui (Tunisia), based on the combined assessment of physical parameters, chemical variables (i.e. nutrients, pigments), sediment pollution and macrobenthic biodiversity. Different port sectors (leisure, fishing, passenger, cargo, shipyard) and different seasons (winter, before touristic period, after touristic period) were compared. Salinity and sediment concentration of copper and antimony were the three environmental parameters most highly correlated with benthic species composition and diversity. Both the environmental variables and the benthic biodiversity patterns were significantly different between the three ports (i.e. different geographical locations). Heraklion port was heavily polluted by AHs in surface and anoxic sediments and had the highest percentage of opportunistic species, while Cagliari had the highest levels of PAHs and UCM and low species richness. El Kantaoui port was less polluted and characterised by a richer biodiversity. The shipyard sector in Heraklion port was significantly different from all other sectors in terms of abiotic and biotic parameters. Physico-chemical and pollution variables recorded during the period after tourism (late summer) were significantly different from the ones recorded in winter. Seasonal differences were not significant between benthic species diversity patterns, but were revealed when the patterns derived from the aggregation of higher taxonomic levels were compared. The present study indicates that a regular-basis monitoring plan including evaluation of environmental health based on benthic biodiversity, can provide a basis for perceiving changes and reveal the degree of anthropogenic disturbance in port environments.

Diverse effects of invasive ecosystem engineers on marine biodiversity and ecosystem functions: A global review and meta-analysis.

Invasive ecosystem engineers (IEE) are potentially one of the most influential types of biological invaders. They are expected to have extensive ecological impacts by altering the physical-chemical structure of ecosystems, thereby changing the rules of existence for a broad range of resident biota. To test the generality of this expectation, we used a global systematic review and meta-analysis to examine IEE effects on the abundance of individual species and communities, biodiversity (using several indices) and ecosystem functions, focusing on marine and estuarine environments. We found that IEE had a significant effect (positive and negative) in most studies testing impacts on individual species, but the overall (cumulative) effect size was small and negative. Many individual studies showed strong IEE effects on community abundance and diversity, but the direction of effects was variable, leading to statistically non-significant overall effects in most categories. In contrast, there was a strong overall effect on most ecosystem functions we examined. IEE negatively affected metabolic functions and primary production, but positively affected nutrient flux, sedimentation and decomposition. We use the results to develop a conceptual model by highlighting pathways whereby IEE impact communities and ecosystem functions, and identify several sources of research bias in the IEE-related invasion literature. Only a few of the studies simultaneously quantified IEE effects on community/diversity and ecosystem functions. Therefore, understanding how IEE may alter biodiversity-ecosystem function relationships should be a primary focus of future studies of invasion biology. Moreover, the clear effects of IEE on ecosystem functions detected in our study suggest that scientists and environmental managers ought to examine how the effects of IEE might be manifested in the services that marine ecosystems provide to humans.

Micro-CTvlab: A web based virtual gallery of biological specimens using X-ray microtomography (micro-CT).

BACKGROUND: During recent years, X-ray microtomography (micro-CT) has seen an increasing use in biological research areas, such as functional morphology, taxonomy, evolutionary biology and developmental research. Micro-CT is a technology which uses X-rays to create sub-micron resolution images of external and internal features of specimens. These images can then be rendered in a three-dimensional space and used for qualitative and quantitative 3D analyses. However, the online exploration and dissemination of micro-CT datasets are rarely made available to the public due to their large size and a lack of dedicated online platforms for the interactive manipulation of 3D data. Here, the development of a virtual micro-CT laboratory (Micro-CTvlab) is described, which can be used by everyone who is interested in digitisation methods and biological collections and aims at making the micro-CT data exploration of natural history specimens freely available over the internet. NEW INFORMATION: The Micro-CTvlab offers to the user virtual image galleries of various taxa which can be displayed and downloaded through a web application. With a few clicks, accurate, detailed and three-dimensional models of species can be studied and virtually dissected without destroying the actual specimen. The data and functions of the Micro-CTvlab can be accessed either on a normal computer or through a dedicated version for mobile devices.

Environmental variability and heavy metal concentrations from five lagoons in the Ionian Sea (Amvrakikos Gulf, W Greece).

BACKGROUND: Coastal lagoons are ecosystems of major importance as they host a number of species tolerant to disturbances and they are highly productive. Therefore, these ecosystems should be protected to ensure stability and resilience. The lagoons of Amvrakikos Gulf form one of the most important lagoonal complexes in Greece. The optimal ecological status of these lagoons is crucial for the well-being of the biodiversity and the economic prosperity of the local communities. Thus, monitoring of the area is necessary to detect possible sources of disturbance and restore stability. NEW INFORMATION: The environmental variables and heavy metals concentrations, from five lagoons of Amvrakikos Gulf were measured from seasonal samplings and compared to the findings of previous studies in the area, in order to check for possible sources of disturbance. The analysis, showed that i) the values of the abiotic parameters vary with time (season), space (lagoon) and with space over time; ii) the variability of the environmental factors and enrichment in certain elements is naturally induced and no source of contamination is detected in the lagoons.

Optimized R functions for analysis of ecological community data using the R virtual laboratory (RvLab).

BACKGROUND: Parallel data manipulation using R has previously been addressed by members of the R community, however most of these studies produce ad hoc solutions that are not readily available to the average R user. Our targeted users, ranging from the expert ecologist/microbiologists to computational biologists, often experience difficulties in finding optimal ways to exploit the full capacity of their computational resources. In addition, improving performance of commonly used R scripts becomes increasingly difficult especially with large datasets. Furthermore, the implementations described here can be of significant interest to expert bioinformaticians or R developers. Therefore, our goals can be summarized as: (i) description of a complete methodology for the analysis of large datasets by combining capabilities of diverse R packages, (ii) presentation of their application through a virtual R laboratory (RvLab) that makes execution of complex functions and visualization of results easy and readily available to the end-user. NEW INFORMATION: In this paper, the novelty stems from implementations of parallel methodologies which rely on the processing of data on different levels of abstraction and the availability of these processes through an integrated portal. Parallel implementation R packages, such as the pbdMPI (Programming with Big Data - Interface to MPI) package, are used to implement Single Program Multiple Data (SPMD) parallelization on primitive mathematical operations, allowing for interplay with functions of the vegan package. The dplyr and RPostgreSQL R packages are further integrated offering connections to dataframe like objects (databases) as secondary storage solutions whenever memory demands exceed available RAM resources. The RvLab is running on a PC cluster, using version 3.1.2 (2014-10-31) on a x86_64-pc-linux-gnu (64-bit) platform, and offers an intuitive virtual environmet interface enabling users to perform analysis of ecological and microbial communities based on optimized vegan functions. A beta version of the RvLab is available after registration at: https://portal.lifewatchgreece.eu/.

Imposex and organotin compounds in ports of the Mediterranean and the Atlantic: Is the story over?

Organotin compounds are toxic substances released into the marine environment from antifouling paints. Sixty-two years following their first application and 9years after the complete ban on their use as biocides in 2003 (EC No. 782/2003), their negative impact on mollusks was still evident, as illustrated by imposex (i.e. the masculinization of female gastropods). This phenomenon is widely used to investigate tributyltin (TBT) pollution, with Hexaplex trunculus and Nassarius nitidus being considered as sensitive bioindicators. H. trunculus specimens and sediment samples collected from the ports of Cagliari (Sardinia), El Kantaoui (Tunisia) and Olhão (Portugal), along with N. nitidus individuals from the port of Faro (Portugal), were used for the assessment of imposex intensity and organotin pollution in these areas. High imposex frequency and organotin concentrations (TBT, triphenyltin (TPhT) and their degradation products) were observed, especially in Sardinia, implying that these chemicals remain a significant pollution issue in the specific Mediterranean ports. Moreover, the highly significant relationship established between penis length of imposex-affected females H. trunculus and TBT/TPhT concentrations offers a rapid and nonsacrificial proxy model for assessing the impact of organotins in coastal environments. Finally, the complementary use of more than one TBT bioindicator is advised in order to obtain more accurate results in detailed monitoring studies of TBT pollution.

Rescuing biogeographic legacy data: The "Thor" Expedition, a historical oceanographic expedition to the Mediterranean Sea.

BACKGROUND: This article describes the digitization of a series of historical datasets based οn the reports of the 1908-1910 Danish Oceanographical Expeditions to the Mediterranean and adjacent seas. All station and sampling metadata as well as biodiversity data regarding calcareous rhodophytes, pelagic polychaetes, and fish (families Engraulidae and Clupeidae) obtained during these expeditions were digitized within the activities of the LifeWatchGreece Research Ιnfrastructure project and presented in the present paper. The aim was to safeguard public data availability by using an open access infrastructure, and to prevent potential loss of valuable historical data on the Mediterranean marine biodiversity. NEW INFORMATION: The datasets digitized here cover 2,043 samples taken at 567 stations during a time period from 1904 to 1930 in the Mediterranean and adjacent seas. The samples resulted in 1,588 occurrence records of pelagic polychaetes, fish (Clupeiformes) and calcareous algae (Rhodophyta). In addition, basic environmental data (e.g. sea surface temperature, salinity) as well as meterological conditions are included for most sampling events. In addition to the description of the digitized datasets, a detailed description of the problems encountered during the digitization of this historical dataset and a discussion on the value of such data are provided.

Macroalgal blooms alter community structure and primary productivity in marine ecosystems.

Eutrophication, coupled with loss of herbivory due to habitat degradation and overharvesting, has increased the frequency and severity of macroalgal blooms worldwide. Macroalgal blooms interfere with human activities in coastal areas, and sometimes necessitate costly algal removal programmes. They also have many detrimental effects on marine and estuarine ecosystems, including induction of hypoxia, release of toxic hydrogen sulphide into the sediments and atmosphere, and the loss of ecologically and economically important species. However, macroalgal blooms can also increase habitat complexity, provide organisms with food and shelter, and reduce other problems associated with eutrophication. These contrasting effects make their overall ecological impacts unclear. We conducted a systematic review and meta-analysis to estimate the overall effects of macroalgal blooms on several key measures of ecosystem structure and functioning in marine ecosystems. We also evaluated some of the ecological and methodological factors that might explain the highly variable effects observed in different studies. Averaged across all studies, macroalgal blooms had negative effects on the abundance and species richness of marine organisms, but blooms by different algal taxa had different consequences, ranging from strong negative to strong positive effects. Blooms' effects on species richness also depended on the habitat where they occurred, with the strongest negative effects seen in sandy or muddy subtidal habitats and in the rocky intertidal. Invertebrate communities also appeared to be particularly sensitive to blooms, suffering reductions in their abundance, species richness, and diversity. The total net primary productivity, gross primary productivity, and respiration of benthic ecosystems were higher during macroalgal blooms, but blooms had negative effects on the productivity and respiration of other organisms. These results suggest that, in addition to their direct social and economic costs, macroalgal blooms have ecological effects that may alter their capacity to deliver important ecosystem services.

Polytraits: A database on biological traits of marine polychaetes.

The study of ecosystem functioning - the role which organisms play in an ecosystem - is becoming increasingly important in marine ecological research. The functional structure of a community can be represented by a set of functional traits assigned to behavioural, reproductive and morphological characteristics. The collection of these traits from the literature is however a laborious and time-consuming process, and gaps of knowledge and restricted availability of literature are a common problem. Trait data are not yet readily being shared by research communities, and even if they are, a lack of trait data repositories and standards for data formats leads to the publication of trait information in forms which cannot be processed by computers. This paper describes Polytraits (http://polytraits.lifewatchgreece.eu), a database on biological traits of marine polychaetes (bristle worms, Polychaeta: Annelida). At present, the database contains almost 20,000 records on morphological, behavioural and reproductive characteristics of more than 1,000 marine polychaete species, all referenced by literature sources. All data can be freely accessed through the project website in different ways and formats, both human-readable and machine-readable, and have been submitted to the Encyclopedia of Life for archival and integration with trait information from other sources.

Engaging the broader community in biodiversity research: the concept of the COMBER pilot project for divers in ViBRANT.

This paper discusses the design and implementation of a citizen science pilot project, COMBER (Citizens' Network for the Observation of Marine BiodivERsity, http://www.comber.hcmr.gr), which has been initiated under the ViBRANT EU e-infrastructure. It is designed and implemented for divers and snorkelers who are interested in participating in marine biodiversity citizen science projects. It shows the necessity of engaging the broader community in the marine biodiversity monitoring and research projects, networks and initiatives. It analyses the stakeholders, the industry and the relevant markets involved in diving activities and their potential to sustain these activities. The principles, including data policy and rewards for the participating divers through their own data, upon which this project is based are thoroughly discussed. The results of the users analysis and lessons learned so far are presented. Future plans include promotion, links with citizen science web developments, data publishing tools, and development of new scientific hypotheses to be tested by the data collected so far.