2D Collagen Membranes from Marine Demosponge Chondrosia reniformis (Nardo, 1847) for Skin-Regenerative Medicine Applications: An In Vitro Evaluation.

Research in tissue engineering and regenerative medicine has an ever-increasing need for innovative biomaterials suitable for the production of wound-dressing devices and artificial skin-like substitutes. Marine collagen is one of the most promising biomaterials for the production of such devices. In this study, for the first time, 2D collagen membranes (2D-CMs) created from the extracellular matrix extract of the marine demosponge Chondrosia reniformis have been evaluated in vitro as possible tools for wound healing. Fibrillar collagen was extracted from a pool of fresh animals and used for the creation of 2D-CMs, in which permeability to water, proteins, and bacteria, and cellular response in the L929 fibroblast cell line were evaluated. The biodegradability of the 2D-CMs was also assessed by following their degradation in PBS and collagenase solutions for up to 21 days. Results showed that C. reniformis-derived membranes avoided liquid and protein loss in the regeneration region and also functioned as a strong barrier against bacteria infiltration into a wound. Gene expression analyses on fibroblasts stated that their interaction with 2D-CMs is able to improve fibronectin production without interfering with the regular extracellular matrix remodeling processes. These findings, combined with the high extraction yield of fibrillar collagen obtained from C. reniformis with a solvent-free approach, underline how important further studies on the aquaculture of this sponge could be for the sustainable production and biotechnological exploitation of this potentially promising and peculiar biopolymer of marine origin.

Photogrammetry as a promising tool to unveil marine caves' benthic assemblages.

Traditionally, monitoring approaches to survey marine caves have been constrained by equipment limitations and strict safety protocols. Nowadays, the rise of new approaches opens new possibilities to describe these peculiar ecosystems. The current study aimed to explore the potential of Structure from Motion (SfM) photogrammetry to assess the abundance and spatial distribution of the sessile benthic assemblages inside a semi-submerged marine cave. Additionally, since impacts of recent date mussel Lithophaga lithophaga illegal fishing were recorded, a special emphasis was paid to its distribution and densities. The results of SfM were compared with a more "traditional approach", by simulating photo-quadrats deployments over the produced orthomosaics. A total of 22 sessile taxa were identified, with Porifera representing the dominant taxa within the cave, and L. lithophaga presenting a density of 88.3 holes/m2. SfM and photo-quadrats obtained comparable results regarding species richness, percentage cover of identified taxa and most of the seascape metrics, while, in terms of taxa density estimations, photo-quadrats highly overestimated their values. SfM resulted in a suitable non-invasive technique to record marine cave assemblages. Seascape indexes proved to be a comprehensive way to describe the spatial pattern of distribution of benthic organisms, establishing a useful baseline to assess future community shifts.

Sponge organic matter recycling: Reduced detritus production under extreme environmental conditions.

Sponges are a key component of coral reef ecosystems and play an important role in carbon and nutrient cycles. Many sponges are known to consume dissolved organic carbon and transform this into detritus, which moves through detrital food chains and eventually to higher trophic levels via what is known as the sponge loop. Despite the importance of this loop, little is known about how these cycles will be impacted by future environmental conditions. During two years (2018 and 2020), we measured the organic carbon, nutrient recycling, and photosynthetic activity of the massive HMA, photosymbiotic sponge Rhabdastrella globostellata at the natural laboratory of Bouraké in New Caledonia, where the physical and chemical composition of seawater regularly change according to the tide. We found that while sponges experienced acidification and low dissolved oxygen at low tide in both sampling years, a change in organic carbon recycling whereby sponges stopped producing detritus (i.e., the sponge loop) was only found when sponges also experienced higher temperature in 2020. Our findings provide new insights into how important trophic pathways may be affected by changing ocean conditions.

Not out of the Mediterranean: Atlantic populations of the gorgonian Paramuricea clavata are a separate sister species under further lineage diversification.

The accurate delimitation of species boundaries in nonbilaterian marine taxa is notoriously difficult, with consequences for many studies in ecology and evolution. Anthozoans are a diverse group of key structural organisms worldwide, but the lack of reliable morphological characters and informative genetic markers hampers our ability to understand species diversification. We investigated population differentiation and species limits in Atlantic (Iberian Peninsula) and Mediterranean lineages of the octocoral genus Paramuricea previously identified as P. clavata. We used a diverse set of molecular markers (microsatellites, RNA-seq derived single-copy orthologues [SCO] and mt-mutS [mitochondrial barcode]) at 49 locations. Clear segregation of Atlantic and Mediterranean lineages was found with all markers. Species-tree estimations based on SCO strongly supported these two clades as distinct, recently diverged sister species with incomplete lineage sorting, P. cf. grayi and P. clavata, respectively. Furthermore, a second putative (or ongoing) speciation event was detected in the Atlantic between two P. cf. grayi color morphotypes (yellow and purple) using SCO and supported by microsatellites. While segregating P. cf. grayi lineages showed considerable geographic structure, dominating circalittoral communities in southern (yellow) and western (purple) Portugal, their occurrence in sympatry at some localities suggests a degree of reproductive isolation. Overall, our results show that previous molecular and morphological studies have underestimated species diversity in Paramuricea occurring in the Iberian Peninsula, which has important implications for conservation planning. Finally, our findings validate the usefulness of phylotranscriptomics for resolving evolutionary relationships in octocorals.

Marine beach litter monitoring strategies along Mediterranean coasts. A methodological review.

Marine beach litter (MBL) represents a serious issue for marine life, coastal ecosystems, human health and several economical activities. The Mediterranean Sea is a semi enclosed basin particularly vulnerable to this problem. Its coasts are threatened by critical anthropogenic pressures that sum up with intensive fishing and shipping, and the slow turnover of its waters. In the last decades, several scientific and participative initiatives have been conducted to study, monitor and clean-up shorelines. These studies were generally characterized by differences in timing and frequency of the surveys, as well as in litter sampling, classification and analysis. This paper presents a systematic review of current literature concerning MBL monitoring strategies along the Mediterranean coasts. Scopus indexed studies are analysed to identify discrepancies and similarities among the applied protocols, understand where current gaps lie, and point out what would be needed to develop a basin-scale efficient monitoring for the Mediterranean Sea.

Complete mitochondrial genome of the branching octocoral Paramuricea grayi (Johnson, 1861), phylogenetic relationships and divergence analysis.

The Gray's sea fan, Paramuricea grayi (Johnson, 1861), typically inhabits deep littoral and circalittoral habitats of the eastern temperate and tropical Atlantic Ocean. Along the Iberian Peninsula, where P. grayi is a dominant constituent of circalittoral coral gardens, two segregating lineages (yellow and purple morphotypes) were recently identified using single-copy nuclear orthologues. The mitochondrial genomes of 9 P. grayi individuals covering both color morphotypes were assembled from RNA-seq data, using samples collected at three sites in southern (Sagres and Tavira) and western (Cape Espichel) Portugal. The complete circular mitogenome is 18,668 bp in length, has an A + T-rich base composition (62.5%) and contains the 17 genes typically found in Octocorallia: 14 protein-coding genes (atp6, atp8, cob, cox1-3, mt-mutS, nad1-6, and nad4L), the small and large subunit rRNAs (rns and rnl), and one transfer RNA (trnM). The mitogenomes were nearly identical for all specimens, though we identified a noteworthy polymorphism (two SNPs 9 bp apart) in the mt-mutS of one purple individual that is shared with the sister species P. clavata. The mitogenomes of the two species have a pairwise sequence identity of 99.0%, with nad6 and mt-mutS having the highest rates of non-synonymous substitutions.

Marine heatwaves drive recurrent mass mortalities in the Mediterranean Sea.

Climate change is causing an increase in the frequency and intensity of marine heatwaves (MHWs) and mass mortality events (MMEs) of marine organisms are one of their main ecological impacts. Here, we show that during the 2015-2019 period, the Mediterranean Sea has experienced exceptional thermal conditions resulting in the onset of five consecutive years of widespread MMEs across the basin. These MMEs affected thousands of kilometers of coastline from the surface to 45 m, across a range of marine habitats and taxa (50 taxa across 8 phyla). Significant relationships were found between the incidence of MMEs and the heat exposure associated with MHWs observed both at the surface and across depths. Our findings reveal that the Mediterranean Sea is experiencing an acceleration of the ecological impacts of MHWs which poses an unprecedented threat to its ecosystems' health and functioning. Overall, we show that increasing the resolution of empirical observation is critical to enhancing our ability to more effectively understand and manage the consequences of climate change.

Mediterranean Sea shelters for the gold coral Savalia savaglia (Bertoloni, 1819): An assessment of potential distribution of a rare parasitic species.

Savalia savaglia is an ecosystem engineer listed as Near-Threatened by the IUCN, even though effective management and proper monitoring efforts to assess its distribution is still lacking. The record of large, long-established colonies can indicate the occurrence of areas with limited human local pressure. These areas may be considered as proxies for the creation of baselines of reference useful to design restoration strategies. The aim of this work was to update the distribution of S. savaglia Mediterranean populations to develop an Ecological Niche Model, highlighting potential areas for future monitoring programs. Occurrence data were collected and harmonized into a single dataset using the scientific literature and validated observations to feed a presence-only MaxEnt model, obtaining a basin-level potential distribution of the species. The results of our study can support decision-makers in marine spatial planning measures including the preservation of mesophotic environments and prioritizing areas for conservation.

Changes in coral forest microbiomes predict the impact of marine heatwaves on habitat-forming species down to mesophotic depths.

Global warming is causing the increase in intensity and frequency of heatwaves, which are often associated with mass mortality events of marine organisms from shallow and mesophotic rocky habitats, including gorgonians and other sessile organisms. We investigated the microbiome responses of the gorgonians Paramuricea clavata, Eunicella cavolini, and the red coral Corallium rubrum to the episodic temperature anomalies detected in the North Western Mediterranean, during August 2011. Although the investigated corals showed no signs of visible necrosis, the abundance of associated Bacteria and Archaea increased with increasing seawater temperature, suggesting their temperature-dependent proliferation. Coral microbiomes were highly sensitive to thermal anomaly amplitude and exhibited increased bacterial diversity to greater thermal shifts. This effect was explained by the decline of dominant bacterial members and the increase of new, rare and opportunistic taxa, including pathogens, revealing a direct effect of heatwave-induced alteration of the microbiomes and not a secondary consequence of coral necrosis.

Scientific knowledge on marine beach litter: A bibliometric analysis.

Litter reduction in the coastal and marine environment represents a major challenge but must be prioritized to preserve biodiversity and ecosystems, as well as the goods and services that humans derive from seas and oceans. This paper reviews the available global scientific literature focusing on marine beach litter and tracks its evolution and trends by combining social network analysis and bibliometrics. The relationships and co-occurrences among authors, countries and keywords retrieved from the Scopus abstract and citation database are presented. A total of 1765 publications are analysed: the majority being journal articles. Results reveal the notable worldwide increase in scientific interest in beach litter in the last decade, as well as its multidisciplinary perspectives. This information could be beneficial for the processes that support the improvement of international efforts for beach litter monitoring, removal, and management activities.

Mediterranean rocky reefs in the Anthropocene: Present status and future concerns.

Global change is striking harder and faster in the Mediterranean Sea than elsewhere, where high levels of human pressure and proneness to climate change interact in modifying the structure and disrupting regulative mechanisms of marine ecosystems. Rocky reefs are particularly exposed to such environmental changes with ongoing trends of degradation being impressive. Due to the variety of habitat types and associated marine biodiversity, rocky reefs are critical for the functioning of marine ecosystems, and their decline could profoundly affect the provision of essential goods and services which human populations in coastal areas rely upon. Here, we provide an up-to-date overview of the status of rocky reefs, trends in human-driven changes undermining their integrity, and current and upcoming management and conservation strategies, attempting a projection on what could be the future of this essential component of Mediterranean marine ecosystems.

Multiple impacts of microplastics can threaten marine habitat-forming species.

Microplastics are recognised as a potential global threat to marine ecosystems, but the biological mechanisms determining their impact on marine life are still largely unknown. Here, we investigated the effects of microplastics on the red coral, a long-lived habitat-forming organism belonging to the Corallium genus, which is present at almost all latitudes from shallow-water to deep-sea habitats. When exposed to microplastics, corals preferentially ingest polypropylene, with multiple biological effects, from feeding impairment to mucus production and altered gene expression. Microplastics can alter the coral microbiome directly and indirectly by causing tissue abrasions that allow the proliferation of opportunistic bacteria. These multiple effects suggest that microplastics at the concentrations present in some marine areas and predicted for most oceans in the coming decades, can ultimately cause coral death. Other habitat-forming suspension-feeding species are likely subjected to similar impacts, which may act synergistically with climate-driven events primarily responsible for mass mortalities.

Synergic effect of global thermal anomalies and local dredging activities on coral reefs of the Maldives.

We investigated possible synergic effects on coral reefs of the local land reclamation activities in the Himmafushi Island (North Malè atoll, Maldives) and the global bleaching event that affected the Maldives in 2016. A BACI (Before-After Control-Impact) sampling design was adopted to contrast effects of dredging activities before and after the occurrence of both dredging and bleaching. The Reef Check protocol, a standardised and worldwide survey method, was applied to collect data through underwater visual surveys on corals, macro-zoobenthos, and fish communities. The bleaching in 2016 hit all the reefs investigated, but only in the reefs around Himmafushi (i.e., the impact sites) the live hard coral reduced significantly its cover and the sand deposited on reefs showed a fourfold increase. Substrate indicators (i.e., coral community and abiotic components) turned out to be more effective than macro-zoobenthos and fish in this short-term environmental impact study.

Sponge microbiome stability during environmental acquisition of highly specific photosymbionts.

In this study, we used in situ transplantations to provide the first evidence of horizontal acquisition of cyanobacterial symbionts by a marine sponge. The acquisition of the symbionts by the host sponge Petrosia ficiformis, which was observed in distinct visible patches, appeared several months after transplantation and at different times on different sponge specimens. We further used 16S rRNA gene amplicon sequencing of genomic DNA (gDNA) and complementary DNA (cDNA) and metatranscriptomics to investigate how the acquisition of the symbiotic cyanobacterium Candidatus Synechococcus feldmannii perturbed the diverse microbiota associated with the host P. ficiformis. To our surprise, the microbiota remained relatively stable during cyanobacterial symbiont acquisition at both structural (gDNA content) and activity (cDNA expression) levels. At the transcriptomic level, photosynthesis was the primary function gained following the acquisition of cyanobacteria. Genes involved in carotene production and oxidative stress tolerance were among those highly expressed by Ca. S. feldmannii, suggesting that this symbiont may protect itself and its host from damaging light radiation.

A high biodiversity mitigates the impact of ocean acidification on hard-bottom ecosystems.

Biodiversity loss and climate change simultaneously threaten marine ecosystems, yet their interactions remain largely unknown. Ocean acidification severely affects a wide variety of marine organisms and recent studies have predicted major impacts at the pH conditions expected for 2100. However, despite the renowned interdependence between biodiversity and ecosystem functioning, the hypothesis that the species' response to ocean acidification could differ based on the biodiversity of the natural multispecies assemblages in which they live remains untested. Here, using experimentally controlled conditions, we investigated the impact of acidification on key habitat-forming organisms (including corals, sponges and macroalgae) and associated microbes in hard-bottom assemblages characterised by different biodiversity levels. Our results indicate that, at higher biodiversity, the impact of acidification on otherwise highly vulnerable key organisms can be reduced by 50 to >90%, depending on the species. Here we show that such a positive effect of a higher biodiversity can be associated with higher availability of food resources and healthy microbe-host associations, overall increasing host resistance to acidification, while contrasting harmful outbreaks of opportunistic microbes. Given the climate change scenarios predicted for the future, we conclude that biodiversity conservation of hard-bottom ecosystems is fundamental also for mitigating the impacts of ocean acidification.

Characterization of North-Western Mediterranean coralligenous assemblages by video surveys and evaluation of their structural complexity.

Thanks to several European directives and conventions there is a general increase of awareness regarding the key ecological role of coralligenous habitats in the Mediterranean Sea, addressing several research projects to standardize protocols for the description of its integrity. Here we surveyed 13 stations along the Italian coasts of the Western Mediterranean Sea, using video-transects technique, comparing the biological structure of coralligenous assemblages and testing the importance of their three-dimensional complexity as a proxy to define their health conditions. We considered the diversity of taxa, fishing impacts and the entity of damage on gorgonian's choenenchyme due to thermal stress, to evidence a gradient in the coralligenous health conditions. Here we developed a method to evaluate coralligenous complexity, selecting categories of taxa particularly sensitive to multiple stressors, named Structural Descriptors to describe the three-dimensional structure of the bioconcretions and to assess a unique Index of 3D - Structural Complexity.

The influence of scuba diving experience on divers' perceptions, and its implications for managing diving destinations.

Scuba diving experience-which can include accumulated diving experience and familiarity with a diving location-is an important descriptor of diver specialisation and behaviour. Formulating and applying generalisations on scuba diving experience and its effects could assist the management of diving destinations around the world. This requires research that tests whether the influences of scuba diving experience are consistent across divers' segments at different locations. The study assessed and compared the influence of scuba diving experience at two study areas in Italy and Mozambique. Scuba divers (N = 499) participated in a survey of diver segmentation, experience, and perceptions. The influence of diving experience on perceptions was determined using canonical correspondence analysis (CCA). Experienced divers provided positive self-assessments, were less satisfied with dive sites' health and management, and viewed the impacts of scuba diving activities less critically than novice divers. Scuba diving experience exerted similar influences on divers, regardless of the study area. However, remarkable differences also emerged between the study areas. Therefore, the use of generalisations on scuba diving experience remains a delicate issue. Recommendations were formulated for the management of experienced scuba diving markets and for the use of generalisations on diving experience to manage diving destinations.

Mediterranean Lithophyllum stictiforme (Corallinales, Rhodophyta) is a genetically diverse species complex: implications for species circumscription, biogeography and conservation of coralligenous habitats.

Lithophyllum species in the Mediterranean Sea function as algal bioconstructors, contributing to the formation of biogenic habitats such as coralligenous concretions. In such habitats, thalli of Lithophyllum, consisting of crusts or lamellae with entire or lobed margins, have been variously referred to as either one species, L. stictiforme, or two species, L. stictiforme and L. cabiochiae, in the recent literature. We investigated species diversity and phylogenetic relationships in these algae by sequencing three markers (psbA and rbcL genes, cox2,3 spacer), in conjunction with methods for algorithmic delimitation of species (ABGD and GMYC). Mediterranean subtidal Lithophyllum belong to a well-supported lineage, hereby called the L. stictiforme complex, which also includes two species described from the Atlantic, L. lobatum and L. searlesii. Our results indicate that the L. stictiforme complex consists of at least 13 species. Among the Mediterranean species, some are widely distributed and span most of the western and central Mediterranean, whereas others appear to be restricted to specific localities. These patterns are interpreted as possibly resulting from allopatric speciation events that took place during the Messinian Salinity Crisis and subsequent glacial periods. A partial rbcL sequence from the lectotype of L. stictiforme unambiguously indicates that this name applies to the most common subtidal Lithophyllum in the central Mediterranean. We agree with recent treatments that considered L. cabiochiae and L. stictiforme conspecific. The diversity of Lithophyllum in Mediterranean coralligenous habitats has been substantially underestimated, and future work on these and other Mediterranean corallines should use identifications based on DNA sequences.