ß-diversity reveals ecological connectivity patterns underlying marine community recovery: Implications for conservation.

As ß-diversity can be seen as a proxy of ecological connections among species assemblages, modeling the decay of similarity in species composition at increasing distance may help elucidate spatial patterns of connectivity and local- to large-scale processes driving community assembly within a marine region. This, in turn, may provide invaluable information for setting ecologically coherent networks of marine protected areas (MPAs) in which protected communities are potentially interrelated and can mutually sustain against environmental perturbations. However, field studies investigating changes in ß-diversity patterns at a range of spatial scales and in relation to disturbance are scant, limiting our understanding of how spatial ecological connections among marine communities may affect their recovery dynamics. We carried out a manipulative experiment simulating a strong physical disturbance on subtidal rocky reefs at several locations spanning >1000 km of coast in the Adriatic Sea (Mediterranean Sea) and compared ß-diversity patterns and decay of similarity with distance and time by current transport between undisturbed and experimentally disturbed macrobenthic assemblages to shed light on connectivity processes and scales involved in recovery. In contrast to the expectation that very local-scale processes, such as vegetative regrowth and larval supply from neighboring undisturbed assemblages, might be the major determinants of recovery in disturbed patches, we found that connectivity mediated by currents at larger spatial scales strongly contributed to shape community reassembly after disturbance. Across our study sites in the Adriatic Sea, ß-diversity patterns suggested that additional protected sites that matched hotspots of propagule exchange could increase the complementarity and strengthen the ecological connectivity throughout the MPA network. More generally, conditional to habitat distribution and selection of sites of high conservation priority (e.g., biodiversity hotspots), setting network internode distance within 100-150 km, along with sizing no-take zones to cover at least 5 km of coast, would help enhance the potential connectivity of Mediterranean subtidal rocky reef assemblages from local to large scale. These results can help improve conservation planning to achieve the goals of promoting ecological connectivity within MPA networks and enhancing their effectiveness in protecting marine communities against rapidly increasing natural and anthropogenic disturbances.

An Alkaloid from a Highly Invasive Seaweed Increases the Voracity and Reproductive Output of a Model Fish Species.

The invasive macroalga Caulerpa cylindracea has spread widely in the Mediterranean Sea, becoming a favorite food item for native fish for reasons yet unknown. By using a combination of behavioral, morphological, and molecular approaches, herein we provide evidence that the bisindole alkaloid caulerpin, a major secondary metabolite of C. cylindracea, significantly increases food intake in the model fish Danio rerio, influencing the regulation of genes involved in the orexigenic pathway. In addition, we found that the compound improves fish reproductive performance by affecting the hypothalamus-pituitary-gonadal axis. The obtained results pave the way for the possible valorization of C. cylindracea as a sustainable source of a functional feed additive of interest to face critical challenges both in aquaculture and in human nutrition.

Short-term physiological and biometrical responses of Lepidium sativum seedlings exposed to PET-made microplastics and acid rain.

Plastics enter in terrestrial natural system primarily by agricultural purposes, while acid rain is the result of anthropogenic activities. The synergistic effects of microplastics and acid rain on plant growth are not known. In this study, different sizes of polyethylene terephthalate (PET) and acid rain are tested on Lepidium sativum, in two separate experimental sets. In the first one we treated plants only with PET, in the second one we used PET and acid rain together. In both experimentations we analyzed: i) plant biometrical parameters (shoot height, leaf number, percentage inhibition of seed germination, fresh biomass), and ii) oxidative stress responses (hydrogen peroxide; ascorbic acid and glutathione). Results carried out from our experiments highlighted that different sizes of polyethylene terephthalate are able to affect plant growth and physiological responses, with or without acid rain supplied during acute toxicity (6 days). SHORT DESCRIPTION: This study showed that different sizes of PET microplastics affect physiological and biometrical responses of Lepidum sativum seedlings, with or without acid rain; roots and leaves responded differently.

¹H NMR Spectroscopy and MVA to Evaluate the Effects of Caulerpin-Based Diet on Diplodus sargus Lipid Profiles.

The biological invasion of the green algae Caulerpa cylindracea represents a serious scientific and public issue in the Mediterranean Sea, essentially due to strong modifications both to habitat structure and native benthic communities. Although alterations in health status and changes in flesh quality of some marine species (dietary exposed to C. cylindracea) have been observed, no studies on cause-effect relationships have been carried out. Here, for the first time, through a controlled feeding experiment followed by ¹H NMR Spectroscopy and multivariate analysis (PCA, OPLS-DA), we showed that caulerpin taken with diet is directly responsible of changes observed in metabolic profile of fish flesh, including alteration of lipid metabolism, in particular with a reduction of ω3 PUFA content. The potential of caulerpin to directly modulate lipid metabolism opens up new questions about causal mechanism triggered by algal metabolite also in view of a possible exploitation in the nutraceutical/medical field.

Fishing for Targets of Alien Metabolites: A Novel Peroxisome Proliferator-Activated Receptor (PPAR) Agonist from a Marine Pest.

Although the chemical warfare between invasive and native species has become a central problem in invasion biology, the molecular mechanisms by which bioactive metabolites from invasive pests influence local communities remain poorly characterized. This study demonstrates that the alkaloid caulerpin (CAU)-a bioactive component of the green alga Caulerpa cylindracea that has invaded the entire Mediterranean basin-is an agonist of peroxisome proliferator-activated receptors (PPARs). Our interdisciplinary study started with the in silico prediction of the ligand-protein interaction, which was then validated by in vivo, ex vivo and in vitro assays. On the basis of these results, we candidate CAU as a causal factor of the metabolic and behavioural disorders observed in Diplodus sargus, a native edible fish of high ecological and commercial relevance, feeding on C. cylindracea. Moreover, given the considerable interest in PPAR activators for the treatment of relevant human diseases, our findings are also discussed in terms of a possible nutraceutical/pharmacological valorisation of the invasive algal biomasses, supporting an innovative strategy for conserving biodiversity as an alternative to unrealistic campaigns for the eradication of invasive pests.

Metabolites from invasive pests inhibit mitochondrial complex II: A potential strategy for the treatment of human ovarian carcinoma?

The red pigment caulerpin, a secondary metabolite from the marine invasive green algae Caulerpa cylindracea can be accumulated and transferred along the trophic chain, with detrimental consequences on biodiversity and ecosystem functioning. Despite increasing research efforts to understand how caulerpin modifies fish physiology, little is known on the effects of algal metabolites on mammalian cells. Here we report for the first time the mitochondrial targeting activity of both caulerpin, and its closely related derivative caulerpinic acid, by using as experimental model rat liver mitochondria, a system in which bioenergetics mechanisms are not altered. Mitochondrial function was tested by polarographic and spectrophotometric methods. Both compounds were found to selectively inhibit respiratory complex II activity, while complexes I, III, and IV remained functional. These results led us to hypothesize that both algal metabolites could be used as antitumor agents in cell lines with defects in mitochondrial complex I. Ovarian cancer cisplatin-resistant cells are a good example of cell lines with a defective complex I function on which these molecules seem to have a toxic effect on proliferation. This provided novel insight toward the potential use of metabolites from invasive Caulerpa species for the treatment of human ovarian carcinoma cisplatin-resistant cells.

1H NMR Spectroscopy and MVA Analysis of Diplodus sargus Eating the Exotic Pest Caulerpa cylindracea.

The green alga Caulerpa cylindracea is a non-autochthonous and invasive species that is severely affecting the native communities in the Mediterranean Sea. Recent researches show that the native edible fish Diplodus sargus actively feeds on this alga and cellular and physiological alterations have been related to the novel alimentary habits. The complex effects of such a trophic exposure to the invasive pest are still poorly understood. Here we report on the metabolic profiles of plasma from D. sargus individuals exposed to C. cylindracea along the southern Italian coast, using 1H NMR spectroscopy and multivariate analysis (Principal Component Analysis, PCA, Orthogonal Partial Least Square, PLS, and Orthogonal Partial Least Square Discriminant Analysis, OPLS-DA). Fish were sampled in two seasonal periods from three different locations, each characterized by a different degree of algal abundance. The levels of the algal bisindole alkaloid caulerpin, which is accumulated in the fish tissues, was used as an indicator of the trophic exposure to the seaweed and related to the plasma metabolic profiles. The profiles appeared clearly influenced by the sampling period beside the content of caulerpin, while the analyses also supported a moderate alteration of lipid and choline metabolism related to the Caulerpa-based diet.

Viral Encephalopathy and Retinopathy in groupers (Epinephelus spp.) in southern Italy: a threat for wild endangered species?

BACKGROUND: Betanodaviruses are the causative agents of Viral Encephalopathy and Retinopathy (VER). To date, more than 50 species have proved to be susceptible and among them, those found in genus Epinephelus are highly represented. Clinical disease outbreaks are generally characterized by typical nervous signs and significant mortalities mainly associated with aquaculture activities, although some concerns for the impact of this infection in wild fish have been raised. In this study, the authors present the first documented report describing an outbreak of VER in wild species in the Mediterranean basin. CASE PRESENTATION: In late summer--early winter 2011 (September-December), significant mortalities affecting wild Dusky grouper (Epinephelus marginatus), Golden grouper (Epinephelus costae) and European sea bass (Dicentrarchus labrax) were reported in the municipality of Santa Maria di Leuca (Northern Ionian Sea, Italy). The affected fish showed an abnormal swimming behavior and swollen abdomens. During this epizootic, five moribund fish showing clear neurological signs were captured and underwent laboratory investigations. Analytical results confirmed the diagnosis of VER in all the specimens. Genetic characterization classified all betanodavirus isolates as belonging to the RGNNV genotype, revealing a close genetic relationship with viral sequences obtained from diseased farmed fish reared in the same area in previous years. CONCLUSION: The close relationship of the viral sequences between the isolates collected in wild affected fish and those isolated during clinical disease outbreaks in farmed fish in the same area in previous years suggests a persistent circulation of betanodaviruses and transmission between wild and farmed stocks. Further investigations are necessary to assess the risk of viral transmission between wild and farmed fish populations, particularly in marine protected areas where endangered species are present.

Parasitic infection in the scyphozoan Rhizostoma pulmo (Macri, 1778).

Very little information is reported for parasites of cnidarians, therefore, the present work aimed to investigate parasitic infections in one of the most widespread jellyfish in the Mediterranean Sea, Rhizostoma pulmo. The goals were to determine prevalence and intensity of parasites in R. pulmo, identify the species involved through morphological and molecular analysis, test whether infection parameters differ in different body parts and in relation to jellyfish size. 58 individuals were collected, 100% of them infected with digenean metacercariae. Intensity varied between 18.7 ± 6.7 per individual in 0-2 cm diameter jellyfish up to 505 ± 50.6 in 14 cm ones. Morphological and molecular analyses suggest that the metacercariae belonged to the family Lepocreadiidae and could be possibly assigned to the genus Clavogalea. Prevalence values of 100% suggest that R. pulmo is an important intermediate host in the life cycle of lepocreadiids in the region. Our findings also support the hypothesis that R. pulmo is an important part in the diet of teleost fish, which are reported as definitive hosts of lepocreadiids, since trophic transmission is necessary for these parasites to complete their life cycles. Parasitological data may therefore be useful to investigate fish-jellyfish predation, integrating traditional methods such as gut contents analysis.

A novel endocast technique providing a 3D quantitative analysis of the gastrovascular system in Rhizostoma pulmo: An unexpected through-gut in cnidaria.

The investigation of jellyfish gastrovascular systems mainly focused on stain injections and dissections, negatively affected by thickness and opacity of the mesoglea. Therefore, descriptions are incomplete and data about tridimensional structures are scarce. In this work, morphological and functional anatomy of the gastrovascular system of Rhizostoma pulmo (Macri 1778) was investigated in detail with innovative techniques: resin endocasts and 3D X-ray computed microtomography. The gastrovascular system consists of a series of branching canals ending with numerous openings within the frilled margins of the oral arms. Canals presented a peculiar double hemi-canal structure with a medial adhesion area which separates centrifugal and centripetal flows. The inward flow involves only the "mouth" openings on the internal wing of the oral arm and relative hemi-canals, while the outward flow involves only the two outermost wings' hemi-canals and relative "anal" openings on the external oral arm. The openings differentiation recalls the functional characteristics of a through-gut apparatus. We cannot define the gastrovascular system in Rhizostoma pulmo as a traditional through-gut, rather an example of adaptive convergence, that partially invalidates the paradigm of a single oral opening with both the uptake and excrete function.

Effects of polyethylene terephthalate (PET) microplastics and acid rain on physiology and growth of Lepidium sativum.

This study evaluated the chronic toxicity (30 days) of different sizes of polyethylene terephthalate (PET) microplastics (60-3000 µm) provided alone or in combination with acid rain, on garden cress (Lepidium sativum). Both biometrical and physiological traits have been evaluated: i) percentage inhibition of seed germination, plant height, leaf number and fresh biomass production; ii) oxidative stress responses (hydrogen peroxide; ascorbic acid and glutathione production); iii) impairment in photosynthetic machinery in term of pigments production; iv) aminolevulinic acid and proline production. Results highlighted that different sizes of PET, alone or in combination with acid rain, are able to negatively affect both biometrical and physiological plant traits. In particular, the lower size of microplastics is able to negatively affect growth and development, as well as to trigger the oxidative burst. Regarding the pigments production, PET coupled with acid rain, induced a higher production of Chl-b, and an inhibition of aminolevulinic acid.

Fishmeal replacement by periphyton reduces the fish in fish out ratio and alimentation cost in gilthead sea bream Sparus aurata.

Aquaculture threatens natural resources by fishing down the sea to supply fishmeal. Alternative protein sources in aquafeeds can provide a solution, particularly those that are waste from other operations and thereby reduce feed production costs. Toward this goal, we examined the waste biomass of marine periphyton from biofilters of an integrated multi-trophic aquaculture (IMTA) system as a replacement for fishmeal in diets of gilthead seabream (Sparus aurata). Four isoproteic (41%) and isolipidic (16.7%) aquafeeds were formulated with increased content of periphyton and a corresponding decrease in fishmeal from 20 to 15, 10, or 0%. The growth and biochemical content of seabream fingerlings (initial body weight 10 g) were examined over 132 days. Replacing 50% of fishmeal by waste periphyton improved feed conversion ratio (1.2 vs. 1.35 in the control diet) without harming fish growth. The complete replacement of fishmeal with periphyton resulted in 15% slower growth but significantly higher protein content in the fish flesh (59 vs. 52% in the control diet). Halving fishmeal content reduced feed cost by US$ 0.13 kg-1 feed and saved 30% in the cost of conversion of feed to fish biomass (US$ 0.58 kg-1 produced fish vs. $0.83 in the control diet). Finally, the total replacement of fishmeal by waste periphyton in the diet reduced the fish in-fish out ratio to below 1 (0.5-0.9) as compared to 1.36 in the control diet. Replacing fishmeal with on-farm produced periphyton minimizes aquaculture footprint through the removal of excess nutrients in effluents and the use of waste biomass to reduce the 'fish in' content in aquafeeds and fish production costs. The present study demonstrates the great practical potential of this dual use of marine periphyton in enhancing the circular economy concept in sustainable fish production.

Phenotypic plasticity under rapid global changes: The intrinsic force for future seagrasses survival.

Coastal oceans are particularly affected by rapid and extreme environmental changes with dramatic consequences for the entire ecosystem. Seagrasses are key ecosystem engineering or foundation species supporting diverse and productive ecosystems along the coastline that are particularly susceptible to fast environmental changes. In this context, the analysis of phenotypic plasticity could reveal important insights into seagrasses persistence, as it represents an individual property that allows species' phenotypes to accommodate and react to fast environmental changes and stress. Many studies have provided different definitions of plasticity and related processes (acclimation and adaptation) resulting in a variety of associated terminology. Here, we review different ways to define phenotypic plasticity with particular reference to seagrass responses to single and multiple stressors. We relate plasticity to the shape of reaction norms, resulting from genotype by environment interactions, and examine its role in the presence of environmental shifts. The potential role of genetic and epigenetic changes in underlying seagrasses plasticity in face of environmental changes is also discussed. Different approaches aimed to assess local acclimation and adaptation in seagrasses are explored, explaining strengths and weaknesses based on the main results obtained from the most recent literature. We conclude that the implemented experimental approaches, whether performed with controlled or field experiments, provide new insights to explore the basis of plasticity in seagrasses. However, an improvement of molecular analysis and the application of multi-factorial experiments are required to better explore genetic and epigenetic adjustments to rapid environmental shifts. These considerations revealed the potential for selecting the best phenotypes to promote assisted evolution with fundamental implications on restoration and preservation efforts.

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.

Global patterns of parasite diversity in cephalopods.

We compiled an updated global catalogue of parasites in cephalopods. Data were used to assess changes in taxonomic distinctness of parasites over two centuries and across the world's oceans, to quantify turnover and nestedness components of parasite ß-diversity, and to attempt estimating their γ-diversity at a global scale. A total of 309 parasites infecting 164 cephalopods were found. We hypothesize that this diversity counts for less than half the potential parasite richness in this molluscan taxon. Taxonomic breadth of parasites was significantly above expectations from null models for Mediterranean Sea and NE Atlantic Ocean, whereas the opposite occurred for NW Pacific Ocean, where a few closely related genera characterized the parasite pool. ß-diversity of parasites was very high and dominated by turnover, except for the Atlantic Ocean where a nested pattern among sub-basins emerged. Taxonomic relatedness of parasite species remained substantially unchanged through time, but species replacements largely occurred over the last two centuries. Our findings highlighted potential hotspots of taxonomic distinctness in cephalopod parasites, geographic regions deserving future research, and the need for a deeper understanding of the magnitude of marine parasite diversity, their biogeography, and their role in marine ecosystems. Our global overview may represent a baseline step for future advances in this direction.

Nanoplastics in the oceans: Theory, experimental evidence and real world.

This review critically analyses >200 papers collected by searching on Pubmed the word "nanoplastics", a group of emerging contaminants which are receiving growing attention. The present review intends to provide an overview of current knowledge on nanoplastic pollution starting with the theory of polymer degradation, passing to laboratory confirmation of nanoplastic formation and ending with the possible occurrence in sea water samples. Most of the observations proposed focus the attention on polystyrene (PS) because the majority of research knowledge is based on this polymer. Moreover, we thoroughly describe what effects have been observed on different organisms tested in controlled conditions. Nanoplastics formation, fate and toxicity seem to be a very dynamic phenomenon. In light of this, we identify some aspects retained crucial when an ecotoxicological study with nanoplastics is performed and which elements of nanoplastics toxicity could be deeper covered.

Exposure to pet-made microplastics: Particle size and pH effects on biomolecular responses in mussels.

This study aims to evaluate the expression of biomarkers of oxidative stress (LPO, GPx, AtCh, SOD) in mussels (Mytilus galloprovincialis) following the exposure to suspensions of microparticles irregular shaped fibres of Polyethylene terephthalate of different sizes (small 5-60 µm, S-PET; medium 61-499 µm, M-PET; large 500-3000 µm, L-PET) at a single dose of 0.1 g/L. Mussels were tested under two different starting pH conditions of marine water: standard (8.0) and acidified (7.5). The results obtained from this study show that: i) PET microplastics are able to induce biochemical stress in mussels; ii) among the biomarkers tested, LPO and GPx were more effective in detecting the stress induced by microplastic in both initial pH conditions; iii) the expression of biomarkers was influenced by the size of the microparticle. In particular, greater effects were associated with the largest PET particle tested (0.5-3.0 mm); iv) regarding the effect of pH, in experiments starting from 7.5 pH the animals showed a lower biomarker expression than those starting from 8.0 pH.

PET microplastics toxicity on marine key species is influenced by pH, particle size and food variations.

This study aims to evaluate effects induced by the exposure of key marine species to leachates and suspensions of different particle-size of PET microparticles, a plastic polymer that is actually considered safe for the environment. Leachates and suspensions of small (5-60 µm); medium (61-499 µm) and large (500-3000 µm) PET were tested on bacteria (V. fischeri; UNI EN ISO 11348-3:2009), algae (P. tricornutum; UNI EN ISO 10253:2016E), and echinoderms (P. lividus; EPA 600/R-95-136/Section 15) species both under standard (8.0) and acidified (7.5) pH conditions. Results obtained show that: i) conversely to larval stage of P. lividus, bacterial and algal tested species are not sensitive to PET pollution under all tested conditions; ii) different tested particle-sizes of PET produce effects that are not always related to their particle-size; iii) differences comparing acidified and standard pH conditions were recorded; iv) concerning echinoderms, food availability produce significant differences compared to fasting conditions; v) special attention on the possible interactions between MPs and other stressors (e.g., food and pH) is needed in order to give a better picture of natural occurring dynamics on marine ecosystems especially in the future frame of global changes.

Local vs regional effects of substratum on early colonization stages of sessile assemblages.

Substratum type and topographic complexity influence the settlement and persistence of benthic organisms. However, the combined effect of these two factors in affecting colonization patterns at different scales has rarely been investigated. A manipulative experiment was conducted to test the interplay of rock type and roughness in affecting the pattern of subtidal assemblages and to provide tests for the generality of effects across a range of spatial scales (centimetres to hundreds of metres). Replicate tiles of four different rock types, with two levels of surface roughness were deployed in rocky subtidal habitats (5 m depth) at two sites (separated by hundreds of metres) at each of three locations (separated by tens of kilometres). Spatial and temporal variation in the colonization patterns over 9 months differed among rock types. However, large-scale processes appeared to be far more important than substratum type or roughness in determining assemblage structure. Predicting the consequences of the introduction of artificial structures into the coastal marine environment is critical as increasingly parts of coastlines are being modified within the Mediterranean and other regions. The results suggest that further investment is needed to manage and mitigate the effects of the deployment of artificial structures in coastal areas.

Are we ready for scaling up restoration actions? An insight from Mediterranean macroalgal canopies.

Extensive loss of macroalgal forests advocates for large-scale restoration interventions, to compensate habitat degradation and recover the associated ecological functions and services. Yet, restoration attempts have generally been limited to small spatial extensions, with the principal aim of developing efficient restoration techniques. Here, the success of outplanting Cystoseira amentacea v. stricta germlings cultured in aquaria was experimentally explored at a scale of tens of kms, by means of a multifactorial experimental design. In the intertidal rocky shores of SE Italy, locations with a continuous distribution for hundreds of meters or with few thalli forming patches of few centimeters of C. amentacea canopy were selected. In each location, the effects of adult conspecifics and the exclusion of macrograzers (salema fish and sea urchins) on the survival of germlings were tested. We evaluated the most critical determinants of mortality for germlings, including the overlooked pressure of mesograzers (e.g. amphipods, small mollusks, polychaetes). Despite the high mortality observed during outplanting and early settlement stages, survival of C. amentacea germlings was consistently favored by the exclusion of macrograzers, while the presence of adult conspecifics had no effects. In addition, the cost analysis of the interventions showed the feasibility of the ex-situ method, representing an essential tool for preserving Cystoseira forests. Large scale restoration is possible but requires baseline information with an in-depth knowledge of the species ecology and of the areas to be restored, together with the development of specific cultivation protocols to make consistently efficient restoration interventions.