Arbacia Lixula Research Articles

Fabrication of antibacterial bone scaffold based on carbonate-substituted hydroxyapatite containing magnesium from black sea urchin (Arbacia lixula) shells reinforced with polyvinyl alcohol and gelatin

Carbonate-substituted hydroxyapatite containing magnesium (Mg–C-HAp) was introduced through dissolution-precipitation treatment of hydroxyapatite containing magnesium (Mg-HAp) based on a black sea urchin (arbacia lixula) shells as novel biogenic materials. Based on chemical composition analysis, the Mg–C-HAp formed A- and B-type CHAp, which contained high carbonate ions. The Ca/P ratio of Mg–C-HAp was 1.707, very close to the biological bone apatite of 1.71. The Mg content in Mg–C-HAp was also relatively high, with the Mg/(Ca + Mg) ratio of 0.139, which is beneficial for antibacterial agents. The morphology of Mg–C-HAp showed particles with nanosize that provide a large surface area of ion promotion. The antibacterial test revealed that the Mg–C-HAp performed high antibacterial activity against Pseudomonas aeruginosa. The Mg–C-HAp-based porous scaffolds were then fabricated using the freeze-drying method with variations of polyvinyl alcohol (PVA) and gelatin fraction. According to the physicochemical analysis, the addition of PVA and gelatin in the scaffold structure decreased the crystallinity of the Mg–C-HAp/PVA/Gel scaffold. This lower crystallinity indicates high biodegradability, which is good for new bone growth. The macropores of the Mg–C-HAp/PVA/Gel scaffold were appropriate for new bone and blood vessel formation. The micropores of the Mg–C-HAp/PVA/Gel scaffold can be a medium for cells to grow. The microporosity of the Mg–C-HAp/PVA/Gel scaffold was also suitable for cell nutrient promotion. The compressive strength of the Mg–C-HAp/PVA/Gel scaffold was sufficient for bone regeneration. The Mg–C-HAp/PVA/Gel scaffold demonstrated high antibacterial activity against P. aeruginosa, so the Mg–C-HAp/PVA/Gel scaffold can maintain the role of Mg and carbonate content for antibacterial purposes. The good physicochemical, mechanical, and antibacterial properties of the Mg–C-HAp/PVA/Gel scaffold represented its suitable characteristics for antibacterial bone scaffolds.

Distribution of sea urchin barrens in shallow algal communities along the eastern Adriatic coast

The sea urchins Paracentrotus lividus and Arbacia lixula are important herbivores in algal communities of the infralittoral rocky bottoms in the Mediterranean Sea. However, grazing by sea urchins that outweighs natural algal recruitment processes might result in the formation of barrens, which are areas dominated by urchins and coralline algae. We present the results of the first large-scale mapping of sea urchin barrens and analyses of sea urchins’ impact on the main algal communities across 1955.5 km of rocky coastline on the eastern side of the Adriatic Sea (Croatia). Since mapping was performed over a geographically wide area and covered a representative quantity of approximately 40% of the coastline of the central part of the Adriatic Sea, the results could be considered to reflect the general situation in that area of the eastern Adriatic coast. Mapping revealed that sea urchin barrens are present along over 35% of the coastline, while the complete absence of algal cover was recorded in 8% of the inspected area. Communities with canopy-forming algae, which are the most valuable shallow-water communities of the Mediterranean Sea, represent the largest proportion of the observed coastline and are affected by sea urchins in over 28% of their extent. Among other factors, the extensive harvesting of date mussels (Lithophaga lithophaga) in the 1970s, 1980s, and early 1990s is likely one of the main activities that resulted in exceptional sea urchin expansion via the removal of micropredators and establishment of shelters for subadult urchins.

Reproductive toxicity assessment of cellulose nanofibers, citric acid, and branched polyethylenimine in sea urchins: Eco-design of nanostructured cellulose sponge framework (Part B)

In the framework of a safe-by-design approach, we previously assessed the eco-safety of nanostructured cellulose sponge (CNS) leachate on sea urchin reproduction. It impaired gamete quality, gamete fertilization competence, and embryo development possibly due to the leaching of chemical additives used during the CNS synthesis process. To extend this observation and identify the component(s) that contribute to CNS ecotoxicity, in the present study, we individually screened the cytotoxic effects on sea urchin Arbacia lixula and Paracentrotus lividus gametes and embryos of the three main constituents of CNS, namely cellulose nanofibers, citric acid, and branched polyethylenimine. The study aimed to minimize any potential safety risk of these components and to obtain an eco-safe CNS. Among the three CNS constituents, branched polyethylenimine resulted in the most toxic agent. Indeed, it affected the physiology and fertilization competence of male and female gametes as well as embryo development in both sea urchin species. These results are consistent with those previously reported for CNS leachate. Moreover, the characterisation of CNS leachate confirmed the presence of detectable branched polyethylenimine in the conditioned seawater even though in a very limited amount. Altogether, these data indicate that the presence of branched polyethylenimine is a cause-effect associated with a significant risk in CNS formulations due to its leaching upon contact with seawater. Nevertheless, the suggested safety protocol consisting of consecutive leaching treatments and conditioning of CNS in seawater can successfully ameliorate the CNS ecotoxicity while maintaining the efficacy of its sorbent properties supporting potential environmental applications.

Evaluating restriction enzyme selection for reduced representation sequencing in conservation genomics.

Conservation genomic studies in non-model organisms generally rely on reduced representation sequencing techniques based on restriction enzymes to identify population structure as well as candidate loci for local adaptation. While the expectation is that the reduced representation of the genome is randomly distributed, the proportion of the genome sampled might depend on the GC content of the recognition site of the restriction enzyme used. Here, we evaluated the distribution and functional composition of loci obtained after a reduced representation approach using Genotyping-by-Sequencing (GBS). To do so, we compared experimental data from two endemic fish species (Symphodus ocellatus and Symphodus tinca, EcoT22I enzyme) and two ecosystem engineer sea urchins (Paracentrotus lividus and Arbacia lixula, ApeKI enzyme). In brief, we mapped the sequenced loci to the phylogenetically closest reference genome available (Labrus bergylta in the fish and Strongylocentrotus purpuratus in the sea urchin datasets), classified them as exonic, intronic and intergenic, and studied their function by using Gene Ontology (GO) terms. We also simulated the effect of using both enzymes in the two reference genomes. In both simulated and experimental data, we detected an enrichment towards exonic or intergenic regions depending on the restriction enzyme used and failed to detect differences between total loci and candidate loci for adaptation in the empirical dataset. Most of the functions assigned to the mapped loci were shared between the four species and involved a myriad of general functions. Our results highlight the importance of restriction enzyme selection and the need for high-quality annotated genomes in conservation genomic studies.

Toxicity of tire particle leachates on early life stages of keystone sea urchin species

Particles from tires are a major fraction of microplastic pollution. They contain a wide range of chemical additives that can leach into the water and be harmful to aquatic organisms. In this study, we investigated the acute toxicity of tire particle leachates in early life stages of three keystone echinoderm species (Paracentrotus lividus, Arbacia lixula, Diadema africanum). Embryos were exposed for 72 h to a range of leachate dilutions, prepared using a concentration of 1 g L−1. Larval growth, abnormal development, and mortality were the measured endpoints. Furthermore, we estimated the activity of glutathione S transferase (GST) and the electron transport system (ETS) in P. lividus. Strong concentration-dependent responses were observed in all species, though with differing sensitivity. The median effect concentrations for abnormal development in P. lividus and A. lixula were 0.16 and 0.35 g L−1, respectively. In D. africanum, mortality overshadowed abnormal development and the median lethal concentration was 0.46 g L−1. Larvae of P. lividus were significantly smaller than the control from 0.125 g L−1, while the other two species were affected from 0.5 g L−1. ETS activity did not change but there was a non-significant trend of increasing GST activity with leachate concentration in P. lividus. Seven organic chemicals and eight metals were detected at elevated concentrations in the leachates. While we regard zinc as a strong candidate to explain some of the observed toxicity, it can be expected that tire particle leachates exhibit a cocktail effect and other leached additives may also contribute to their toxicity. Our results emphasize the importance of multi-species studies as they differ in their susceptibility to tire particle pollution. We found negative effects at concentrations close to projections in the environment, which calls for more research and mitigation actions on these pollutants.

Echinoderm larvae as bioindicators for the assessment of marine pollution: Sea urchin and sea cucumber responsiveness and future perspectives

Echinoderms play a crucial role in the functioning of marine ecosystems and due to their extensive distribution, rapid response, and the high sensitivity of their planktonic larvae to a large range of stressors, some species are widely used as biological indicators. In addition to sea urchins, sea cucumbers have recently been implemented in embryotoxicity bioassays showing high potential in ecotoxicological studies. However, the use of this species is still hindered by a lack of knowledge regarding their comparative responsiveness. The present study aimed to investigate the responsiveness of different echinoderm species to environmental pollution in order to develop their integration in batteries of ecotoxicological bioassays. To this end, the embryos of two sea urchins (Paracentrotus lividus and Arbacia lixula) and two sea cucumbers (Holothuria polii and Holothuria tubulosa) were incubated with inorganic and organic toxicants (cadmium, copper, mercury, lead, sodium dodecyl sulphate and 4-n-Nonhyphenol) and elutriates from contaminated marine sediments, chosen as a case study model. The results obtained, expressed through the percentage of abnormal embryos and Integrative Toxicity Indices (ITI), indicated species-specific sensitivities to pollutants, with comparable and correlated responsiveness between sea urchins and sea cucumbers. More specifically, sea cucumber larvae exposed to elutriates appear to be more sensitive than sea urchins, especially when incubated with samples containing trace metals, PCB and TBT. These results indicate that toxic responses in embryos exposed to environmental matrices are probably modulated by interactions between different variables, including additive, synergistic and antagonistic effects. These findings suggest that performing a larval test using different echinoderm classes can integrate the interactive effects of bioavailable fraction of contaminants on various levels, providing sensitive, representative and all year-round batteries of bioassays to apply in ecotoxicological studies.

In Vitro Anti-Inflammatory and Vasculoprotective Effects of Red Cell Extract from the Black Sea Urchin Arbacia lixula.

Sea urchins have emerged as an important source of bioactive compounds with anti-inflammatory and antioxidant properties relevant to human health. Since inflammation is a crucial pathogenic process in the development and progression of atherosclerosis, we here assessed the potential anti-inflammatory and vasculoprotective effects of coelomic red-cell methanolic extract of the black sea urchin Arbacia lixula in an in vitro model of endothelial cell dysfunction. Human microvascular endothelial cells (HMEC-1) were pretreated with A. lixula red-cell extract (10 and 100 μg/mL) before exposure to the pro-inflammatory cytokine tumor necrosis factor (TNF)-α. The extract was non-toxic after 24 h cell treatment and was characterized by antioxidant power and phenol content. The TNF-α-stimulated expression of adhesion molecules (VCAM-1, ICAM-1) and cytokines/chemokines (MCP-1, CCL-5, IL-6, IL-8, M-CSF) was significantly attenuated by A. lixula red-cell extract. This was functionally accompanied by a reduction in monocyte adhesion and chemotaxis towards activated endothelial cells. At the molecular level, the tested extract significantly counteracted the TNF-α-stimulated activation of the pro-inflammatory transcription factor NF-κB. These results provide evidence of potential anti-atherosclerotic properties of A. lixula red-cell extract, and open avenues in the discovery and development of dietary supplements and/or drugs for the prevention or treatment of cardiovascular diseases.

Polymer Chemical Identity as a Key Factor in Microplastic-Insecticide Antagonistic Effects during Embryogenesis of Sea Urchin Arbacia lixula.

As a proxy for pollutants that may be simultaneously present in urban wastewater streams, the effects of two microplastics-polystyrene (PS; 10, 80 and 230 μm diameter) and polymethylmethacrylate (PMMA; 10 and 50 μm diameter)-on fertilisation and embryogenesis in the sea urchin Arbacia lixula with co-exposure to the pyrethroid insecticide cypermethrin were investigated. Synergistic or additive effects were not seen for plastic microparticles (50 mg L-1) in combination with cypermethrin (10 and 1000 μg L-1) based on evaluation of skeletal abnormalities or arrested development and death of significant numbers of larvae during the embryotoxicity assay. This behaviour was also apparent for male gametes pretreated with PS and PMMA microplastics and cypermethrin, where a reduction in sperm fertilisation ability was not evidenced. However, a modest reduction in the quality of the offspring was noted, suggesting that there may be some transmissible damage to the zygotes. PMMA microparticles were more readily taken up than PS microparticles, which could suggest surface chemical identity as potentially modulating the affinity of larvae for specific plastics. In contrast, significantly reduced toxicity was noted for the combination of PMMA microparticles and cypermethrin (100 μg L-1), and may be related to less ready desorption of the pyrethroid than PS, as well as cypermethrin activating mechanisms that result in reduced feeding and hence decreased ingestion of microparticles.

Influence of Salinity on Copper Toxicity in Paracentrotus lividus and Arbacia lixula Embryos

The sea urchins Paracentrotus lividus and Arbacia lixula have a key role in benthic dynamics and they are often used to assess the toxicity of environmental contaminants. The aim of this research was to investigate the influence of salinity on copper toxicity on P. lividus and A. lixula embryos, evaluating the impact of future ocean salinity variations and estimating an optimum range for embryos. For this purpose, sea urchin embryos were exposed to different copper concentrations under varying salinity regimes, estimating the toxic effects with the percentage of abnormal embryos and the Integrative Toxicity Index (ITI). Results revealed acceptable levels of anomalies in a salinity range of 33–36‰ for A. lixula and 33–39‰ for P. lividus. The effect concentrations (EC50) of copper reveals increasing toxicity at increasing salinity for both sea urchin species, but between 33–36‰ (A. lixula) and 36–39‰ (P. lividus) the toxicity appear inversely related with salinity. This study illustrates the difficulty in relating a biological response to bioavailability, because the uptake and the sensitivity to copper across salinities is influenced by differences in osmoregulation, enhancing our knowledge about optimal salinity ranges and the potential impact of climate change on sea urchin embryo development.

Breaking the paradigm: Marine sediments hold two-fold microplastics than sea surface waters and are dominated by fibers

We conducted one of the first studies to integrate the quantification and characterization of microplastics (MPs), including fibers, in different habitats (sea surface, seafloor and beach sediments) of a coastal Mediterranean marine protected area, analyzing their ingestion in several marine species. The objectives of the study were to evaluate the distribution of MPs according to shape and polymer, to assess the contribution of fibers to local plastic pollution and to evaluate their ingestion in fish and invertebrates species that inhabit the study area (Pagrus pagrus, Serranus scriba, Spondyliosoma cantharus, Diplodus vulgaris, Oblada melanura, Holothuria forskalii, Holothuria tubularis, Holothuria polis, Arbacia lixula, Paracentrotus lividus, Modiolus barbatus, Mytilus galloprovincialis and Arca noae). A total of 111 environmental samples were analyzed. The mean abundance of MPs (excluding fibers) quantified in beach sediments (13,418.86 ± 28,787.99 MPs/m2) was two orders of magnitude higher than that found in seafloor sediments (76.92 ± 108.84 MPs/m2), which in turn was two orders of magnitude higher than sea surface samples (0.17 ± 0.39 MPs/m2). The fibers were the most abundant shape of MPs identified in all habitats. Variability in MPs ingestion was detected between species, with ingestion rates ranging from 43 % to 100 % for general MPs and ranging from 7 % to 100 % for fibers. The highest ingestion was observed in Holoturians, representing suitable bioindicators for plastic pollution. The composition of the polymer varies weakly depending on habitats and biota, but the result is strongly correlated with the morphology of the plastic. Fibers were mainly composed of cellulose acetate (29 %), styrofoam of polystyrene (18 %), and filaments, films and fragments of polyethylene and polypropylene. The results highlighted the need to expand integrated approaches to effectively study marine plastic pollution and to undertake efficient actions to limit the input of plastics, particularly fibers, into the marine environment.

Effects of Sulfamethoxazole on Fertilization and Embryo Development in the Arbacia lixula Sea Urchin.

Simple SummaryDrugs released into the aquatic environment create serious problems for the organisms that live there. For this reason, the present study investigates the in vitro effects of the antibiotic sulfamethoxazole, widely found in wastewater, on the fertilization and development of the Arbacia lixula sea urchin. The results showed a significant reduction in the percentage of fertilized oocytes at the highest drug concentrations, together with an increase in anomalies and delays in the development of the embryo. Therefore, the data obtained suggest urgent intervention on the release of these drugs in order to prevent important alterations in the species’ development and to preserve biodiversity.To date, drugs released into the aquatic environment are a real problem, and among antibiotics, sulfamethoxazole is the one most widely found in wastewater; thus, the evaluation of its toxicity on marine organisms is very important. This study, for the first time, investigates the in vitro effects of 4 concentrations of sulfamethoxazole (0.05 mg/L, 0.5 mg/L, 5 mg/L, 50 mg/L) on the fertilization and development of the sea urchin Arbacia lixula. The gametes were exposed to drugs in three different stages: simultaneously with, prior to, and post-fertilization. The results show a significant reduction in the percentage of fertilized oocytes at the highest drug concentrations. Moreover, an increase in anomalies and delays in embryo development following the treatment with the drug was demonstrated. Therefore, the data suggest that this antibiotic can alter the development of marine organisms, making it urgent to act to reduce their release and to determine the concentration range with the greatest impact.

Biological and Proteomic Characterization of the Anti-Cancer Potency of Aqueous Extracts from Cell-Free Coelomic Fluid of Arbacia lixula Sea Urchin in an In Vitro Model of Human Hepatocellular Carcinoma

Echinoderms are an acknowledged source of bioactive compounds exerting various beneficial effects on human health. Here, we examined the potential in vitro anti-hepatocarcinoma effects of aqueous extracts of the cell-free coelomic fluid obtained from the sea urchin Arbacia lixula using the HepG2 cell line as a model system. This was accomplished by employing a combination of colorimetric, microscopic and flow cytometric assays to determine cell viability, cell cycle distribution, the possible onset of apoptosis, the accumulation rate of acidic vesicular organelles, mitochondrial polarization, cell redox state and cell locomotory ability. The obtained data show that exposed HepG2 cells underwent inhibition of cell viability with impairment of cell cycle progress coupled to the onset of apoptotic death, the induction of mitochondrial depolarization, the inhibition of reactive oxygen species production and acidic vesicular organelle accumulation, and the block of cell motile attitude. We also performed a proteomic analysis of the coelomic fluid extract identifying a number of proteins that are plausibly responsible for anti-cancer effects. Therefore, the anti-hepatocarcinoma potentiality of A. lixula’s preparation can be taken into consideration for further studies aimed at the characterization of the molecular mechanism of cytotoxicity and the development of novel prevention and/or treatment agents.

Comparative multi-species analysis of potassium cyanide toxicity

Potassium cyanide (KCN), a highly water soluble and bioaccumulative cyanide salt, is examined to determine the toxic effects by using two green algae (Dunaliella viridis, Nannochloropsis oculata) and genetically different two sea urchin (Paracentrotus lividus, Arbacia lixula) species. To determine the toxic effects on the early developmental stages of sea urchin embryos, 72-hour embryotoxicity studies were conducted. Potassium cyanide toxicity at cellular level was also investigated and 6-hour embryos of both sea urchin species were used to determine genotoxic effects of KCN. Since plutei naturally feed on microalgae, two species of plankton were used to reveal phytotoxic effects of KCN. KCN was found to be embryo- geno- and phytotoxic. EC50's for P. lividus and A. lixula were found 7.96 and 6.52 μM. IC50's for N. oculata for 48 h and 72 h were found 23.66 and 80.45 μM. IC50's for D. viridis for 48 h and 72 h were found 14.31 and 23.36 μM.

Marine rocky reef assemblages and lithological properties of substrates are connected at different ecological levels

Recent studies suggest that the lithological properties of hard substrates play an important role in influencing the diversity and structure of marine assemblages involving macroalgae, sessile organisms and vagile animals like heterobranchs gastropods and fishes. The present study aims at exploring whether the influence of different substrates (limestone vs granite) could drive the occurrence of different vagile groups (crustacean decapods, echinoderms and fishes). The study was carried out at the “Tavolara-Punta Coda Cavallo” Marine Protected Area (NE Sardinia, Mediterranean Sea) where substrates of different lithology, namely granites and limestones, occur on a local spatial scale. The diversity and the abundance of 16 vagile species (four crustaceans, seven echinoderms and five fishes) were investigated by the analysis of photographs (2352 images) collected in 20 sites (10 limestones and 10 granites), between 30 and 50 m depth. Statistical analyses showed a clear-cut separation between the assemblages assessed on the two lithological substrate types, with assemblages on granites that were tightly grouped and those on limestones more dispersed. The total species richness did not significantly vary between limestones and granites. Galathea strigosa and Marthasterias glacialis were exclusively recorded on granites, while Palinurus elephas, Scyllarides latus, Arbacia lixula, Ophidiaster ophidianus and Serranus scriba were exclusively found on limestones. The observed patterns could be partially explained by multiple factors related to lithology: i) the different availability of preferred food sources, potentially influenced by substrate type; ii) the site geomorphology, that could provide different dens and refuges (in terms of quantity and types of shelters) suitable for different species; iii) the colour of different substrates enhancing the mimicry of different species according to their livery and camouflage ability. Our and literature data suggest that the substrate type in rocky reefs could interact with other environmental factors (i.e., light intensity related to depth) in shaping the structure of rocky-reef assemblages.

Effects of diclofenac on the gametes and embryonic development of Arbacia lixula

Diclofenac (DCF), a non-steroidal anti-inflammatory drug (NSAID), is among the main pollutants of aquatic environments. Nevertheless, even if several authors evaluated its effects on marine organisms, no work has ever analysed its impact on the sea urchin Arbacia lixula. The purpose of this study was to analyse, for the first time, the impact of DCF at different concentrations (50 mg/l, 5 mg/l, 0.5 mg/l, 0.05 mg/l) on A. lixula embryos and gametes to indicate possible safe minimum levels of release of this drug in marine environments. Our results showed significant concentration-dependent effects on embryonic development (decrease in the number of developed embryos and increase in the quantity of degenerated eggs) and significant levels of bioaccumulation at DCF concentrations of 50 mg/l in both gametes and embryos. DCF showed a significant impact on the fertilisation of A. lixula due to its effects on gametes, confirming the possibility of environmental risk and highlighting the need to improve wastewater treatment and drug disposal processes.

Influence of Winter Storms on the Sea Urchin Pathogen Assemblages

In recent years, recurrent sea urchin mass mortalities in the Canary Islands have been registered. These mortality-related events have decimated 93% of the eastern Atlantic populations of the barren-forming sea urchin Diadema africanum. Two severe episodes of rough southeastern seas led to winter storms in February 2010 (Xynthia) and February 2018 (Emma) and preceded the last mass mortality event. We hypothesized that these events are related to the mass mortalities registered during the February in those years. Previous studies identified Neoparamoeba branchiphila as the causal agent of the disease, possibly acting in synergy with Vibrio alginolyticus and/or other pathogens. To determine the link between winter storms and the sea urchin pathogen community, we monitored the marine pathogen assemblage before and after the winter storm Filomena (February 2020) on Tenerife Island, on different habitats (sea water, sediment and algae) and in four species of sea urchin hosts (D. africanum, Arbacia lixula, Paracentrotus lividus and Sphaerechinus granularis). A total of six pathogens, including N. branchiphila, Vexillifera minutissima, Acanthamoeba sp., Vahlkampfia sp., V. alginolyticus and green colonies of Vibrio spp., were identified. Only small amoebas were found in sea urchins, while Vibrio species were more common in seawater, sediment and algae substrates. V. alginolyticus was occasionally detected in three sea urchins specimens, while N. branchiphila was found in the coelom of all four sea urchin studied. As previously hypothesized, a significant pathogen increment in seawater and in the sea urchin species D. africanum and P. lividus, was found after Filomena. Our results confirmed the relationship between the winter storms and marine pathogen dynamics. However, further studies are needed to demonstrate the direct relationship between these pathogen increases and the sea urchin mass mortalities.

The Two Sides of the Mediterranean: Population Genomics of the Black Sea Urchin Arbacia lixula (Linnaeus, 1758) in a Warming Sea

Global environmental changes may have a profound impact on ecosystems. In this context, it is crucial to gather biological and ecological information of the main species in marine communities to predict and mitigate potential effects of shifts in their distribution, abundance, and interactions. Using genotyping by sequencing (GBS), we assessed the genetic structure of a keystone species in the Mediterranean shallow littoral ecosystems, the black sea urchin Arbacia lixula. This bioengineer species can shape their communities due to its grazing activity and it is experiencing an ongoing expansion with increasing temperatures. The population genomic analyses on 5,241 loci sequenced in 240 individuals from 11 Mediterranean sampled populations revealed that all populations were diverse and showed significant departure from equilibrium. Albeit genetic differentiation was in general shallow, a significant break separated the western and eastern Mediterranean populations, a break not detected in previous studies with less resolutive markers. Notably, no clear effect of the Almería-Oran front, an important break in the Atlanto-Mediterranean transition, could be detected among the western basin populations, where only a slight differentiation of the two northernmost populations was found. Despite the generally low levels of genetic differentiation found, we identified candidate regions for local adaptation by combining different genomic analysis with environmental data. Salinity, rather than temperature, seemed to be an important driver of genetic structure in A. lixula. Overall, from a population genomics standpoint, there is ample scope for A. lixula to continue thriving and adapting in the warming Mediterranean.

Boron originating from produced water discharged to Brazilian coastal waters: Ecological risk assessment with marine organisms

Deterministic and probabilistic ecological risk assessments were performed for the boron present in coastal waters in the region of the São Sebastião channel (coast of São Paulo State, Brazil) surrounding the outfall of the São Sebastião waterway terminal (TEBAR) and in reference areas far from the outfall. A set of ecotoxicity tests with 9 marine organisms (Lytechinus variegatus, Arbacia lixula, Skeletonema costatum, Asterionellopsis glacialis, Parablenius pilicornis, Artemia salina, Megabalanus coccopoma, Mysidopsis juniae and Hypnea musciformes) was performed in the laboratory. Ecotoxicity tests confirmed that boron presents low ecotoxicity, with Lytechinus variegatus being the most sensitive species studied, with an EC50 of 14.6 mg L−1 and a no-observed effect concentration (NOEC) of 6.75 mg L−1. According to the deterministic ecological risk assessment, no significant environmental impact is expected if we consider the most sensitive of the organisms tested and the highest concentration of boron found in the coastal waters (5.82 mg L−1).

Fertilization and development of Arbacia lixula eggs are affected by osmolality conditions

The sea urchin Arbacia lixula coexist with Paracentrotus lividus in the Mediterranean, but the two sea urchin species are quite different from each other. Concerning the female gamete, A. lixula eggs are much darker than those of P. lividus due to the characteristic pigmentation. Upon insemination, the fertilization envelope formed by A. lixula eggs is remarkably thinner than that of P. livius eggs, which implies that the cortical organization of the eggs in the two species may be quite different. In this communication, we examined the phenotypic plasticity of A. lixula eggs in the changing osmolality. The plasma membrane, cortical actin cytoskeleton and vesicles are extensively altered in the eggs exposed to 40% seawater for 15 min. When fertilized, the Ca2+ response in these eggs was significantly compromised and the sperm often failed to enter the eggs. Remarkably, the pattern of the Ca2+ response was restored when these eggs were transferring back to the natural seawater before fertilization, while the actin cytoskeleton partially reverted to the original state. Nonetheless, these eggs restored in seawater failed to regain the innate sperm receptivity that allows only one sperm to enter in natural seawater. Thus, the ability to guide monospermic fertilization is lost by water entry into the eggs, and the eggs incorporated either multiple or no sperm. On the other hand, eggs briefly exposed to hypertonic seawater exhibited no evident morphological anomaly. Nonetheless, the monospermic eggs that experienced a brief exposure (15 min) to hypertonic seawater prior to fertilization in natural seawater displayed a subtly altered sperm-induced Ca2+ response and morpho-functional anomaly around the pluteus stage. Our results suggest that A. lixula eggs attain only a limited extent of cytological plasticity, and that the osmolality shock affects the physical nature of the egg surface which in turn affects the developmental programming.

Interpopulation variability of the reproductive cycle of Arbacia lixula (Echinodermata: Echinoidea) in the Mostaganem shallow-water area (south-western Mediterranean)

The study on the reproductive biology of the black sea urchin Arbacia lixula was carried out at two different sites on the west coast of Algeria (Mostaganem) (one located at Salamandre considered polluted and the other at Stidia is relatively healthy). A monthly sampling was carried out from January 2015 to December 2016. The sex ratio at the two sites is slightly in favour of males. Monthly monitoring of the gonad index showed maximum values during the month of May, and minimum values in September. The gonad index for both male and female evolves in the same way. Histological study of the male and the female gonads confirmed the results obtained for the gonad index. The sexual resting stage takes place in winter, followed by maturation in spring. Finally, the spawning phase occurs between spring and summer. The variability of the seasonal temperature values appears to influence the maturation of the gonads and the spawning process in A. lixula. The results obtained in this study show the existence of an annual cycle in A.lixula with the presence of a massive spawning in July, the duration of which depends on the combined effect of stress factors (increase in temperature and presence of pollution).