Microbial diversity in four Mediterranean irciniid sponges.

This paper describes a dataset of microbial communities from four different sponge species: Irciniaoros (Schmidt, 1864), Irciniavariabilis (Schmidt, 1862), Sarcotragusspinosulus Schmidt, 1862 and Sarcotragusfasciculatus (Pallas, 1766). The examined sponges all belong to Demospongiae (Class); Keratosa (Subclass); Dictyoceratida (Order); Irciniidae (Family). Samples were collected by scuba diving at depths between 6-14 m from two sampling sites of rocky formations at the northern coast of Crete (Cretan Sea, eastern Mediterranean) and were subjected to metabarcoding for the V5-V6 region of the 16S rRNA gene.

Comparative Chemical Profiling and Antimicrobial/Anticancer Evaluation of Extracts from Farmed versus Wild Agelas oroides and Sarcotragus foetidus Sponges.

Marine sponges are highly efficient in removing organic pollutants and their cultivation, adjacent to fish farms, is increasingly considered as a strategy for improving seawater quality. Moreover, these invertebrates produce a plethora of bioactive metabolites, which could translate into an extra profit for the aquaculture sector. Here, we investigated the chemical profile and bioactivity of two Mediterranean species (i.e., Agelas oroides and Sarcotragus foetidus) and we assessed whether cultivated sponges differed substantially from their wild counterparts. Metabolomic analysis of crude sponge extracts revealed species-specific chemical patterns, with A. oroides and S. foetidus dominated by alkaloids and lipids, respectively. More importantly, farmed and wild explants of each species demonstrated similar chemical fingerprints, with the majority of the metabolites showing modest differences on a sponge mass-normalized basis. Furthermore, farmed sponge extracts presented similar or slightly lower antibacterial activity against methicillin-resistant Staphylococcus aureus, compared to the extracts resulting from wild sponges. Anticancer assays against human colorectal carcinoma cells (HCT-116) revealed marginally active extracts from both wild and farmed S. foetidus populations. Our study highlights that, besides mitigating organic pollution in fish aquaculture, sponge farming can serve as a valuable resource of biomolecules, with promising potential in pharmaceutical and biomedical applications.

Uptake of aquaculture-related dissolved organic pollutants by marine sponges: Kinetics and mechanistic insights from a laboratory study.

Besides the release of organic matter from uneaten feed and fish excreta, a considerable amount of deleterious chemicals may also end up into the marine environment from intensive aquaculture. A fraction of these pollutants remains freely dissolved and pose a threat to marine life due to increased bioavailability. Given the filter-feeding ability of sponges, we investigated the capacity of four ubiquitous Mediterranean species (Agelas oroides, Axinella cannabina, Chondrosia reniformis and Sarcotragus foetidus) in removing aquaculture-related dissolved organic pollutants. These included individual chemicals belonging to antibiotics (i.e., oxytetracycline), antifouling biocides (i.e., diuron and Irgarol 1051) and polycyclic aromatic hydrocarbons (i.e., 2,6-dimethylnapththalene, phenanthrene). The uptake of pollutants was assessed in vitro by exposing small sponge explants to each chemical for a period of 8 h. Additional "cleanup" experiments were performed for complex mixtures mimicking the dissolved organic material encountered in fish farms, such as filtrates of fish feed and excreta. All sponges exhibited a pronounced preference for lipophilic pollutants and a strong positive correlation was revealed between clearance rate and substrate hydrophobicity. Our best filter-feeder (i.e., A. oroides) was able to clear 10.0 ± 1.3 mL of seawater per hour and per gram of sponge, when exposed to 2,6-dimethylnapththalene. Active pumping was found to be the predominant mechanism dictating the assimilation of dissolved pollutants in all sponge species, as it was 3-10 times faster than pollutants' passive adsorption on sponges' pinacoderm. Additionally, the uptaken pollutants were shown to be strongly retained by sponges and they were hardly released back to seawater as a result of desorption or sponge excretory mechanisms. Our study corroborates that sponges are highly efficient in uptaking dissolved organic compounds and it offers new insights into the kinetics and mechanisms ruling this process.

Shift and homogenization of gut microbiome during invasion in marine fishes.

Biological invasion is one of the main components of global changes in aquatic ecosystems. Unraveling how establishment in novel environments affects key biological features of animals is a key step towards understanding invasion. Gut microbiome of herbivorous animals is important for host health but has been scarcely assessed in invasive species. Here, we characterized the gut microbiome of two invasive marine herbivorous fishes (Siganus rivulatus and Siganus luridus) in their native (Red Sea) and invaded (Mediterranean Sea) ranges. The taxonomic and phylogenetic diversity of the microbiome increased as the fishes move away from the native range and its structure became increasingly different from the native microbiome. These shifts resulted in homogenization of the microbiome in the invaded range, within and between the two species. The shift in microbial diversity was associated with changes in its functions related with the metabolism of short-chain fatty acids. Altogether, our results suggest that the environmental conditions encountered by Siganidae during their expansion in Mediterranean ecosystems strongly modifies the composition of their gut microbiome along with its putative functions. Further studies should pursue to identify the precise determinants of these modifications (e.g. changes in host diet or behavior, genetic differentiation) and whether they participate in the ecological success of these species.

A Multi-Species Investigation of Sponges' Filtering Activity towards Marine Microalgae.

Chronic discharge of surplus organic matter is a typical side effect of fish aquaculture, occasionally leading to coastal eutrophication and excessive phytoplankton growth. Owing to their innate filter-feeding capacity, marine sponges could mitigate environmental impact under integrated multitrophic aquaculture (IMTA) scenarios. Herein, we investigated the clearance capacity of four ubiquitous Mediterranean sponges (Agelas oroides, Axinella cannabina, Chondrosia reniformis and Sarcotragus foetidus) against three microalgal substrates with different size/motility characteristics: the nanophytoplankton Nannochloropsis sp. (~3.2 µm, nonmotile) and Isochrysis sp. (~3.8 µm, motile), as well as the diatom Phaeodactylum tricornutum (~21.7 µm, nonmotile). In vitro cleaning experiments were conducted using sponge explants in 1 L of natural seawater and applying different microalgal cell concentrations under light/dark conditions. The investigated sponges exhibited a wide range of retention efficiencies for the different phytoplankton cells, with the lowest average values found for A. cannabina (37%) and the highest for A. oroides (70%). The latter could filter up to 14.1 mL seawater per hour and gram of sponge wet weight, by retaining 100% of Isochrysis at a density of 105 cells mL-1, under darkness. Our results highlight differences in filtering capacity among sponge species and preferences for microalgal substrates with distinct size and motility traits.

A de novo transcriptome assembly for the bath sponge Spongia officinalis, adjusting for microsymbionts.

OBJECTIVES: We report a transcriptome acquisition for the bath sponge Spongia officinalis, a non-model marine organism that hosts rich symbiotic microbial communities. To this end, a pipeline was developed to efficiently separate between bacterial expressed genes from those of eukaryotic origin. The transcriptome was produced to support the assessment of gene expression and, thus, the response of the sponge, to elevated temperatures, replicating conditions currently occurring in its native habitat. DATA DESCRIPTION: We describe the assembled transcriptome along with the bioinformatic pipeline used to discriminate between signals of metazoan and prokaryotic origin. The pipeline involves standard read pre-processing steps and incorporates extra analyses to identify and filter prokaryotic reads out of the analysis. The proposed pipeline can be followed to overcome the technical RNASeq problems characteristic for symbiont-rich metazoan organisms with low or non-existent tissue differentiation, such as sponges and cnidarians. At the same time, it can be valuable towards the development of approaches for parallel transcriptomic studies of symbiotic communities and the host.

Comparative Evaluation of Essential Oils from Medicinal-Aromatic Plants of Greece: Chemical Composition, Antioxidant Capacity and Antimicrobial Activity against Bacterial Fish Pathogens.

The administration of antibiotics in aquaculture has raised concern about the impact of their overuse in marine ecosystems, seafood safety and consumers' health. This "green consumerism" has forced researchers to find new alternatives against fish pathogens. The present study focused on 12 Mediterranean medicinal-aromatic plants as potential antimicrobials and antioxidant agents that could be used in fish aquaculture. In vitro assays showed that the essential oils (EOs) from all studied plants had anti-bacterial and antioxidant properties, with their efficacy being dependent on their chemical composition. More specifically, EOs rich in carvacrol, p-cymene and γ-terpinene exhibited not only the strongest inhibitory activity against the growth of bacterial pathogens (inhibitory concentration: 26-88 µg mL-1), but also the greatest total antioxidant capacity (ABTS: 2591-5879 µmole mL-1; CUPRAC: 931-2733 µmole mL-1). These compounds were mainly found in the EOs from Greek oregano (Origanum vulgare subsp. hirtum), Spanish oregano (Thymbra capitata) and savoury (Satureja thymbra) collected from cultivations in Greece. The specific EOs stand out as promising candidates for the treatment of bacterial diseases and oxidative stress in farmed fish. Further in vivo experiments are needed to fully understand the effects of EO dietary supplementation on fish farming processes.

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.

The sponge microbiome project.

Marine sponges (phylum Porifera) are a diverse, phylogenetically deep-branching clade known for forming intimate partnerships with complex communities of microorganisms. To date, 16S rRNA gene sequencing studies have largely utilised different extraction and amplification methodologies to target the microbial communities of a limited number of sponge species, severely limiting comparative analyses of sponge microbial diversity and structure. Here, we provide an extensive and standardised dataset that will facilitate sponge microbiome comparisons across large spatial, temporal, and environmental scales. Samples from marine sponges (n = 3569 specimens), seawater (n = 370), marine sediments (n = 65) and other environments (n = 29) were collected from different locations across the globe. This dataset incorporates at least 268 different sponge species, including several yet unidentified taxa. The V4 region of the 16S rRNA gene was amplified and sequenced from extracted DNA using standardised procedures. Raw sequences (total of 1.1 billion sequences) were processed and clustered with (i) a standard protocol using QIIME closed-reference picking resulting in 39 543 operational taxonomic units (OTU) at 97% sequence identity, (ii) a de novo clustering using Mothur resulting in 518 246 OTUs, and (iii) a new high-resolution Deblur protocol resulting in 83 908 unique bacterial sequences. Abundance tables, representative sequences, taxonomic classifications, and metadata are provided. This dataset represents a comprehensive resource of sponge-associated microbial communities based on 16S rRNA gene sequences that can be used to address overarching hypotheses regarding host-associated prokaryotes, including host specificity, convergent evolution, environmental drivers of microbiome structure, and the sponge-associated rare biosphere.

CIGESMED for divers: Establishing a citizen science initiative for the mapping and monitoring of coralligenous assemblages in the Mediterranean Sea.

BACKGROUND: Over the last decade, inventorying and monitoring of marine biodiversity has significantly benefited from the active engagement of volunteers. Although several Citizen Science projects concern tropical reef ecosystems worldwide, none of the existing initiatives has yet specifically focused on their Mediterranean equivalents. Mediterranean coralline reefs, known as "coralligenous", are bioherms primarily built by calcifying rhodophytes on hard substrates under dim-light conditions; they are considered hotspots of biodiversity and are extremely popular among divers due to their complex structure, conspicuous biological wealth and high aesthetic value. Nevertheless, data on their distribution, structure and conservation status is lacking for several Mediterranean areas while they are vulnerable to an increasing number of threats. NEW INFORMATION: In the framework of CIGESMED SeasEra (ERAnet) project a specialized Citizen Science project was launched, aiming to engage enthusiast divers in the study and monitoring of Mediterranean coralligenous assemblages through the gathering of basic information regarding their spatial occurrence, assemblage structure and associated pressures or threats. For its active implementation, a data collection protocol and a multilingual website were developed, comprising an educational module and a data submission platform. Georeferenced 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. A variety of tools is provided to facilitate end users, while divers have the choice to report additional information and are encouraged to upload their photographs. The long-term goal is the development of an active community of amateur observers providing widespread and ecologically significant data on coralligenous assemblages.

Testing the robustness of Citizen Science projects: Evaluating the results of pilot project COMBER.

BACKGROUND: Citizen Science (CS) as a term implies a great deal of approaches and scopes involving many different fields of science. The number of the relevant projects globally has been increased significantly in the recent years. Large scale ecological questions can be answered only through extended observation networks and CS projects can support this effort. Although the need of such projects is apparent, an important part of scientific community cast doubt on the reliability of CS data sets. NEW INFORMATION: The pilot CS project COMBER has been created in order to provide evidence to answer the aforementioned question in the coastal marine biodiversity monitoring. The results of the current analysis show that a carefully designed CS project with clear hypotheses, wide participation and data sets validation, can be a valuable tool for the large scale and long term changes in marine biodiversity pattern change and therefore for relevant management and conservation issues.

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.

The biodiversity of the Mediterranean Sea: estimates, patterns, and threats.

The Mediterranean Sea is a marine biodiversity hot spot. Here we combined an extensive literature analysis with expert opinions to update publicly available estimates of major taxa in this marine ecosystem and to revise and update several species lists. We also assessed overall spatial and temporal patterns of species diversity and identified major changes and threats. Our results listed approximately 17,000 marine species occurring in the Mediterranean Sea. However, our estimates of marine diversity are still incomplete as yet-undescribed species will be added in the future. Diversity for microbes is substantially underestimated, and the deep-sea areas and portions of the southern and eastern region are still poorly known. In addition, the invasion of alien species is a crucial factor that will continue to change the biodiversity of the Mediterranean, mainly in its eastern basin that can spread rapidly northwards and westwards due to the warming of the Mediterranean Sea. Spatial patterns showed a general decrease in biodiversity from northwestern to southeastern regions following a gradient of production, with some exceptions and caution due to gaps in our knowledge of the biota along the southern and eastern rims. Biodiversity was also generally higher in coastal areas and continental shelves, and decreases with depth. Temporal trends indicated that overexploitation and habitat loss have been the main human drivers of historical changes in biodiversity. At present, habitat loss and degradation, followed by fishing impacts, pollution, climate change, eutrophication, and the establishment of alien species are the most important threats and affect the greatest number of taxonomic groups. All these impacts are expected to grow in importance in the future, especially climate change and habitat degradation. The spatial identification of hot spots highlighted the ecological importance of most of the western Mediterranean shelves (and in particular, the Strait of Gibraltar and the adjacent Alboran Sea), western African coast, the Adriatic, and the Aegean Sea, which show high concentrations of endangered, threatened, or vulnerable species. The Levantine Basin, severely impacted by the invasion of species, is endangered as well. This abstract has been translated to other languages (File S1).