Photoprotective and Anti-Aging Properties of the Apical Frond Extracts from the Mediterranean Seaweed Ericaria amentacea.

There is a growing interest in using brown algal extracts thanks to the bioactive substances they produce for adaptation to the marine benthic environment. We evaluated the anti-aging and photoprotective properties of two types of extracts (50%-ethanol and DMSO) obtained from different portions, i.e., apices and thalli, of the brown seaweed, Ericaria amentacea. The apices of this alga, which grow and develop reproductive structures during summer when solar radiation is at its peak, were postulated to be rich in antioxidant compounds. We determined the chemical composition and pharmacological effects of their extracts and compared them to the thallus-derived extracts. All the extracts contained polyphenols, flavonoids and antioxidants and showed significant biological activities. The hydroalcoholic apices extracts demonstrated the highest pharmacological potential, likely due to the higher content of meroditerpene molecular species. They blocked toxicity in UV-exposed HaCaT keratinocytes and L929 fibroblasts and abated the oxidative stress and the production of pro-inflammatory cytokines, typically released after sunburns. Furthermore, the extracts showed anti-tyrosinase and anti-hydrolytic skin enzyme activity, counteracting the collagenase and hyaluronidase degrading activities and possibly slowing down the formation of uneven pigmentation and wrinkles in aging skin. In conclusion, the E. amentacea apices derivatives constitute ideal components for counteracting sunburn symptoms and for cosmetic anti-aging lotions.

The role of habitat in the facilitation of Ostreopsis spp. blooms.

In recent decades, recurrent Ostreopsis spp. blooms have been recorded throughout the globe, causing public health issues and mass mortalities of invertebrates. Ostreopsis species are benthic and develop in shallow waters in close relation with a substrate, but possible substrate preferences are still ambiguous. Bloom develops on both living and dead substrates and several interacting biotic and abiotic factors acting at different spatial scales can potentially foster or regulate Ostreopsis spp. development. The objective of this review is to collect and summarize information on Ostreopsis spp. blooms related to the habitat at different spatial scales, in order to assess preferences and trends. References including Ostreopsis spp. samplings in the field were analysed in this review, as potentially including information about the micro- (substrate), meso­ (community) and macrohabitat (ecosystem) related to Ostreopsis spp. blooms. The sampled substrate and the ecosystem where Ostreopsis spp. were collected were generally reported and described in the studies, while the description of the mesohabitat was rarely reported. Ostreopsis spp. were generally described as attached to biotic substrates and in particular, macroalgae, even in studies conducted in coral reefs, where macroalgae are generally not dominant (but they can be in case of coral reef degradation). In both temperate and tropical areas, Ostreopsis spp. were mostly sampled on algal species usually forming medium or low complexity communities (erect or turf-forming algae), often characteristic from post-regime shift scenarios, and rarely on canopy-forming species (such as fucoids and kelps). This literature review highlights the need of collecting more information about the mesohabitat where important Ostreopsis spp. blooms develop, as much as of the underlying mechanisms driving eventual differences on Ostreopsis spp. abundances. This knowledge would allow a better risk assessment of Ostreopsis spp. blooms, identifying areas at high risk on the base of the benthic habitats.

An integrated assessment of the Good Environmental Status of Mediterranean Marine Protected Areas.

Local, regional and global targets have been set to halt marine biodiversity loss. Europe has set its own policy targets to achieve Good Environmental Status (GES) of marine ecosystems by implementing the Marine Strategy Framework Directive (MSFD) across member states. We combined an extensive dataset across five Mediterranean ecoregions including 26 Marine Protected Areas (MPAs), their reference unprotected areas, and a no-trawl case study. Our aim was to assess if MPAs reach GES, if their effects are local or can be detected at ecoregion level or up to a Mediterranean scale, and which are the ecosystem components driving GES achievement. This was undertaken by using the analytical tool NEAT (Nested Environmental status Assessment Tool), which allows an integrated assessment of the status of marine systems. We adopted an ecosystem approach by integrating data from several ecosystem components: the seagrass Posidonia oceanica, macroalgae, sea urchins and fish. Thresholds to define the GES were set by dedicated workshops and literature review. In the Western Mediterranean, most MPAs are in good/high status, with P. oceanica and fish driving this result within MPAs. However, GES is achieved only at a local level, and the Mediterranean Sea, as a whole, results in a moderate environmental status. Macroalgal forests are overall in bad condition, confirming their status at risk. The results are significantly affected by the assumption that discrete observations over small spatial scales are representative of the total extension investigated. This calls for large-scale, dedicated assessments to realistically detect environmental status changes under different conditions. Understanding MPAs effectiveness in reaching GES is crucial to assess their role as sentinel observatories of marine systems. MPAs and trawling bans can locally contribute to the attainment of GES and to the fulfillment of the MSFD objectives. Building confidence in setting thresholds between GES and non-GES, investing in long-term monitoring, increasing the spatial extent of sampling areas, rethinking and broadening the scope of complementary tools of protection (e.g., Natura 2000 Sites), are indicated as solutions to ameliorate the status of the basin.

Myelin sheath and cyanobacterial thylakoids as concentric multilamellar structures with similar bioenergetic properties.

There is a surprisingly high morphological similarity between multilamellar concentric thylakoids in cyanobacteria and the myelin sheath that wraps the nerve axons. Thylakoids are multilamellar structures, which express photosystems I and II, cytochromes and ATP synthase necessary for the light-dependent reaction of photosynthesis. Myelin is a multilamellar structure that surrounds many axons in the nervous system and has long been believed to act simply as an insulator. However, it has been shown that myelin has a trophic role, conveying nutrients to the axons and producing ATP through oxidative phosphorylation. Therefore, it is tempting to presume that both membranous structures, although distant in the evolution tree, share not only a morphological but also a functional similarity, acting in feeding ATP synthesized by the ATP synthase to the centre of the multilamellar structure. Therefore, both molecular structures may represent a convergent evolution of life on Earth to fulfill fundamentally similar functions.

The Remarkable Antioxidant and Anti-Inflammatory Potential of the Extracts of the Brown Alga Cystoseira amentacea var. stricta.

Inflammation and oxidative stress are part of the complex biological responses of body tissues to harmful stimuli. In recent years, due to the increased understanding that oxidative stress is implicated in several diseases, pharmaceutical industries have invested in the research and development of new antioxidant compounds, especially from marine environment sources. Marine seaweeds have shown the presence of many bioactive secondary metabolites, with great potentialities from both the nutraceutical and the biomedical point of view. In this study, 50%-ethanolic and DMSO extracts from the species C. amentacea var. stricta were obtained for the first time from seaweeds collected in the Ligurian Sea (north-western Mediterranean). The bioactive properties of these extracts were then investigated, in terms of quantification of specific antioxidant activities by relevant ROS scavenging spectrophotometric tests, and of anti-inflammatory properties in LPS-stimulated macrophages by evaluation of inhibition of inflammatory cytokines and mediators. The data obtained in this study demonstrate a strong anti-inflammatory effect of both C. amentacea extracts (DMSO and ethanolic). The extracts showed a very low grade of toxicity on RAW 264.7 macrophages and L929 fibroblasts and a plethora of antioxidant and anti-inflammatory effects that were for the first time thoroughly investigated. The two extracts were able to scavenge OH and NO radicals (OH EC50 between 392 and 454 µg/mL; NO EC50 between 546 and 1293 µg/mL), to partially rescue H2O2-induced RAW 264.7 macrophages cell death, to abate intracellular ROS production in H2O2-stimulated macrophages and fibroblasts and to strongly inhibit LPS-induced inflammatory mediators, such as NO production and IL-1α, IL-6, cyclooxygenase-2 and inducible NO synthase gene expression in RAW 264.7 macrophages. These results pave the way, for the future use of C. amentacea metabolites, as an example, as antioxidant food additives in antiaging formulations as well as in cosmetic lenitive lotions for inflamed and/or damaged skin.

Erratum to "A new record of the invasive seaweed Caulerpa cylindracea Sonder in the South Adriatic Sea" [Heliyon 5 (9), (September 2019), e02449].

[This corrects the article DOI: 10.1016/j.heliyon.2019.e02449.].

A new record of the invasive seaweed Caulerpa cylindracea Sonder in the South Adriatic Sea.

The green alga Caulerpa cylindracea Sonder is one of the most infamous and threatening invasive species in the Mediterranean Sea. Since 1985, it started rapidly spreading to all Mediterranean regions causing many ecological changes on natural communities. In the present study, we present an example of this proliferation with the first record in the Marine Protected Area of Tremiti Island (MPATI) in the South Adriatic Sea. Fifteen sites along the coast and 5 different depths have been investigated. Our results provide eveidence of a wide invasion of this pest in three islands, San Domino, San Nicola and Capraia. This study fills a particular data gap in the ongoing biomonitoring of invasive seaweeds in the Mediterranean Sea representing a base line of this invasive species for the MPATI.

First ex situ outplanting of the habitat-forming seaweed Cystoseira amentacea var. stricta from a restoration perspective.

In the Mediterranean Sea, brown algae belonging to the Cystoseira genus play a valuable role as foundation species. Due to evidences of regression/loss of the habitats of these species caused by the interplay of human and climatic disturbances, active restoration measures have been encouraged by EU regulations. In particular, nondestructive restoration techniques, which avoid the depletion of threatened species in donor populations, are strongly recommended. In the framework of the EU project ROCPOP-Life, the first ex situ outplanting experience of Cystoseira amentacea var. stricta has been implemented in the Cinque Terre Marine Protected Area (northwestern Mediterranean). A total of 400 clay tiles, hosting approximately three mm-long germlings of C. amentacea, were fixed to the rocky shore with screws: the tiles were monitored for the next 2 months by photographic sampling, and survival (presence/absence of juveniles on the tiles), cover and growth were assessed. Additional sampling was performed 6 months after tile deployment, after which an unprecedented storm surge severely affected the restoration performance. After 2 months, over 40% of the tiles were covered with Cystoseira juveniles, which reached approximately eight mm in total length. The tiles that survived the storm hosted three to six cm-long juveniles. The high cover (≥25%), assuring moisture and shading, and the appropriate size of the juveniles, to avert micro-grazing, at time of deployment were key to the survival and growth of the outplanted juveniles, increasing the potential for restoration success. Our findings show that outplanting of midlittoral canopy-forming species is a feasible approach for restoration efforts, with particular attention given to the early phases: (i) laboratory culture, (ii) transport, and (iii) juvenile densities. These results are strongly encouraging for the implementation of restoration actions for C. amentacea on a large scale, in light of EU guidelines.

Robustness of Adamussium colbecki shell to ocean acidification in a short-term exposure.

Atmospheric pCO2 has increased since the industrial revolution leading to a lowering of the ocean surface water pH, a phenomenon called ocean acidification (OA). OA is claimed to be a major threat for marine organisms and ecosystems and, particularly, for Polar regions. We explored the impact of OA on the shell mechanical properties of the Antarctic scallop Adamussium colbecki exposed for one month to acidified (pH 7.6) and natural conditions (unmanipulated littoral water), by performing Scanning Electron Microscopy, nanoindentation and Vickers indentation on the scallop shell. No effect of pH could be detected either in crystal deposition or in the mechanical properties. A. colbecki shell was found to be resistant to OA, which suggests this species to be able to face a climate change scenario that may threat the persistence of the endemic Antarctic species. Further investigation should be carried out in order to elucidate the destiny of this key species in light of global change.

The impact of ocean acidification on the gonads of three key Antarctic benthic macroinvertebrates.

CO2 atmospheric pressure is increasing since industrial revolution, leading to a lowering of the ocean surface water pH, a phenomenon known as ocean acidification, with several reported effects on individual species and cascading effects on marine ecosystems. Despite the great amount of literature on ocean acidification effects on calcifying organisms, the response of their reproductive system still remains poorly known. In the present study, we investigated the histopathological effects of low pH on the gonads of three key macroinvertebrates of the Terra Nova Bay (Ross Sea) littoral area: the sea urchin Sterechinus neumayeri, the sea star Odontaster validus and the scallop Adamussium colbecki. After 1 month of exposure at control (8.12) and reduced (7.8 and 7.6) pH levels, we dissected the gonads and performed histological analyses to detect potential differences among treatments. Results showed significant effects on reproductive conditions of A. colbecki and S. neumayeri, while O. validus did not show any kind of alteration. Present results reinforce the need to focus on ocean acidification effects on soft tissues, particularly the gonads, whose damage may exert large effects on the individual fitness, with cascading effects on the population dynamic of the species.

The multiple roles of ß-diversity help untangle community assembly processes affecting recovery of temperate rocky shores.

Metacommunity theory highlights the potential of ß-diversity as a useful link to empirical research, especially in diverse systems where species exhibit a range of stage-dependent dispersal characteristics. To investigate the importance of different components and scales of ß-diversity in community assembly, we conducted a large-scale disturbance experiment and compared relative recovery across multiple sites and among plots within sites on the rocky shore. Six sites were spread along 80 km of coastline and, at each site, five plots were established, matching disturbed and undisturbed quadrats. Recovery was not complete at any of the sites after 1 year for either epibenthos (mostly composed of macroalgae and, locally, mussels) or infauna. Significant differences in recovery among sites were observed for epibenthos but not for infauna, suggesting that different community assembly processes were operating. This was supported by epibenthos in the recovering plots having higher species turnover than in undisturbed sediment, and recovery well predicted by local diversity, while infaunal recovery was strongly influenced by the epibenthic community's habitat complexity. However, infaunal community recovery did not simply track formation of habitat by recovering epibenthos, but appeared to be overlain by within-site and among-site aspects of infaunal ß-diversity. These results suggest that documenting changes in the large plants and animals alone will be a poor surrogate for rocky shore community assembly processes. No role for ecological connectivity (negative effect of among-site ß-diversity) in driving recovery was observed, suggesting a low risk of effects from multiple disturbances propagating along the coast, but a limited resilience at the site scale to large-scale disturbances such as landslides or oil spills.

Ex situ cultivation protocol for Cystoseira amentacea var. stricta (Fucales, Phaeophyceae) from a restoration perspective.

Due to multiple impacts, Cystoseira forests are experiencing a significant decline, which is affecting the ecosystem services they provide. Despite conservation efforts, there is an urgent need to develop best practices and large-scale restoration strategies. To implement restoration actions, we developed an ex situ protocol for the cultivation of Cystoseira. amentacea var. stricta, aimed at reducing the time needed for laboratory culture, thus avoiding prolonged maintenance and minimizing costs. Specifically, we tested the effects of temperature, light and substratum on settlement and growth of early life stages using a factorial experiment. Temperature (20 and 24°C) and photoperiod (15L:9D) were selected to reflect the conditions experienced in the field during the reproductive period. Two light intensities (125 and 250 µmol photons m-2s-1) were selected to mimic the condition experienced in the absence of canopy (i.e. barren-higher light intensity) or in the understory (lower light intensity) during gamete release. The tested substrata were flat polished pebbles and rough clay tiles. The release of gametes and the successive survival and development of embryo and germlings were followed for two weeks. Regardless of the culture conditions, rougher tiles showed higher zygote settlement, but the substrata did not affect the successive development. Zygote mortality after one week averaged 50% and at the end of the second week, embryonic survival was higher under lower light and temperature conditions, which also determined the growth of larger embryos.

Ecological status improvement over a decade along the Ligurian coast according to a macroalgae based index (CARLIT).

According to the Water Framework Directive, within 2015 European Union countries must reach and maintain the "good" Ecological Status (ES), quantified through indices based on key biological elements as indicators. Along the Ligurian shallow rocky coasts (NW Italy), a macroalgae based index (CARtography of LITtoral and upper-sublittoral benthic communities, CARLIT), calibrated according to national characteristics and management needs, has been applied by the Regional Environmental Agency over the last ten years. In 2015, at least a "good" ES was achieved in all Ligurian water bodies except one, located in the Eastern Ligurian coastline, characterized by the lack of the most sensitive species, Cystoseira amentacea var. stricta. A general ES improvement has been observed along the Ligurian coastline, also in comparison with other quality indices (macroinvertebrates and fecal bacteria), and in particular in the Genoa water body, as proved by a relevant increase of C. amentacea abundance, probably as a consequence of enhancement in wastewater treatments. In the present study, the reliability of the observed improvement of the ES over a decade has been assessed, teasing apart intra-seasonal and operator-related variability. These results support the reliability of monitoring procedures carried out though the CARLIT Index and highlight the need and the effectiveness of reduction measures for anthropogenic impacts in order to achieve the ES required by European directives.

Management of harmful benthic dinoflagellates requires targeted sampling methods and alarm thresholds.

Concern regarding Benthic Harmful Algal Blooms (BHABs) is increasing since some harmful benthic species have been identified in new areas. In the Mediterranean basin, the most common harmful benthic microalgae are Ostreopsis cf. ovata and Prorocentrum lima, which produce palytoxin-like compounds and okadaic acid respectively, and the need to implement monitoring activities has increased. However, a general agreement on appropriate strategies (e.g. sampling season, definition of alarm thresholds, etc.) is still lagging behind, especially for P. lima, whose proliferation dynamics are still poorly known.

Population genomics meet Lagrangian simulations: Oceanographic patterns and long larval duration ensure connectivity among Paracentrotus lividus populations in the Adriatic and Ionian seas.

Connectivity between populations influences both their dynamics and the genetic structuring of species. In this study, we explored connectivity patterns of a marine species with long-distance dispersal, the edible common sea urchin Paracentrotus lividus, focusing mainly on the Adriatic-Ionian basins (Central Mediterranean). We applied a multidisciplinary approach integrating population genomics, based on 1,122 single nucleotide polymorphisms (SNPs) obtained from 2b-RAD in 275 samples, with Lagrangian simulations performed with a biophysical model of larval dispersal. We detected genetic homogeneity among eight population samples collected in the focal Adriatic-Ionian area, whereas weak but significant differentiation was found with respect to two samples from the Western Mediterranean (France and Tunisia). This result was not affected by the few putative outlier loci identified in our dataset. Lagrangian simulations found a significant potential for larval exchange among the eight Adriatic-Ionian locations, supporting the hypothesis of connectivity of P. lividus populations in this area. A peculiar pattern emerged from the comparison of our results with those obtained from published P. lividus cytochrome b (cytb) sequences, the latter revealing genetic differentiation in the same geographic area despite a smaller sample size and a lower power to detect differences. The comparison with studies conducted using nuclear markers on other species with similar pelagic larval durations in the same Adriatic-Ionian locations indicates species-specific differences in genetic connectivity patterns and warns against generalizing single-species results to the entire community of rocky shore habitats.

A novel application of an adaptable modeling approach to the management of toxic microalgal bloom events in coastal areas.

Harmful algal blooms have been increasing in frequency in recent years, and attention has shifted from describing to modeling and trying to predict these phenomena, since in many cases they pose a risk to human health and coastal activities. Predicting ecological phenomena is often time and resource consuming, since a large number of field collected data are required. We propose a novel approach that involves the use of modeled meteorological data as input features to predict the concentration of the toxic benthic dinoflagellate Ostreopsis cf. ovata in seawater. Ten meteorological features were used to train a Quantile Random Forests model, which was then validated using field collected concentration data over the course of a summer sampling season. The proposed model was able to accurately describe Ostreopsis abundance in the water column in response to meteorological variables. Furthermore, the predictive power of this model appears good, as indicated by the validation results, especially when the quantile for predictions is tuned to match management requirements. The Quantile Random Forests method was selected, as it allows for greater flexibility in the generated predictions, thus making this model suitable as a tool for coastal management. The application of this approach is novel, as no other models or tools that are adaptable to this degree are currently available. The model presented here was developed for a single species over a limited geographical extension, but its methodological basis appears flexible enough to be applied to the prediction of HABs in general and it could also be extended to the case of other ecological phenomena that are strongly dependent on meteorological drivers, that can be independently modeled and potentially globally available.

Ostreopsis fattorussoi sp. nov. (Dinophyceae), a new benthic toxic Ostreopsis species from the eastern Mediterranean Sea.

The new benthic toxic dinoflagellate, Ostreopsis fattorussoi sp. nov., is described from the Eastern Mediterranean Sea, Lebanon and Cyprus coasts, and is supported by morphological and molecular data. The plate formula, Po, 3', 7″, 6c, 7s, 5‴, 2'''', is typical for the Ostreopsis genus. It differs from all other Ostreopsis species in that (i) the curved suture between plates 1' and 3' makes them approximately hexagonal, (ii) the 1' plate lies in the left half of the epitheca and is obliquely orientated leading to a characteristic shape of plate 6″. The round thecal pores are bigger than the other two Mediterranean species (O. cf. ovata and O. cf. siamensis). O. fattorussoi is among the smallest species of the genus (DV: 60.07 ± 5.63 µm, AP: 25.66 ± 2.97 µm, W: 39.81 ± 5.05 µm) along with O. ovata. Phylogenetic analyses based on the LSU and internal transcribed spacer rDNA shows that O. fattorussoi belongs to the Atlantic/Mediterranean Ostreopsis spp. clade separated from the other Ostreopsis species. Ostreopsis fattorussoi produces OVTX-a and structural isomers OVTX-d and -e, O. cf. ovata is the only other species of this genus known to produce these toxins. The Lebanese O. fattorussoi did not produce the new palytoxin-like compounds (ovatoxin-i, ovatoxin-j1 , ovatoxin-j2 , and ovatoxin-k) that were previously found in O. fattorussoi from Cyprus. The toxin content was in the range of 0.28-0.94 pg · cell-1 . On the Lebanon coast, O. fattorussoi was recorded throughout the year 2015 (temperature range 18°C-31.5°C), with peaks in June and August.

Microelectrode array (MEA) platform as a sensitive tool to detect and evaluate Ostreopsis cf. ovata toxicity.

In the last decade, the occurrence of harmful dinoflagellate blooms of the genus Ostreopsis has increased both in frequency and in geographic distribution with adverse impacts on public health and the economy. Ostreopsis species are producers of palytoxin-like toxins (putative palytoxin and ovatoxins) which are among the most potent natural non-protein compounds known to date, exhibiting extreme toxicity in mammals, including humans. Most existing toxicological data are derived from in vivo mouse assay and are related to acute effects of pure palytoxin, without considering that the toxicity mechanism of dinoflagellates can be dependent on the varying composition of complex biotoxins mixture and on the presence of cellular components. In this study, in vitro neuronal networks coupled to microelectrode array (MEA)-based system are proposed, for the first time, as sensitive biosensors for the evaluation of marine alga toxicity on mammalian cells. Toxic effect was investigated by testing three different treatments of laboratory cultured Ostreopsis cf. ovata cells: filtered and re-suspended algal cells; filtered, re-suspended and sonicated algal cells; conditioned growth medium devoid of algal cells. The great sensitivity of this system revealed the mixture of PTLX-complex analogues naturally released in the growth medium and the different potency of the three treatments to inhibit the neuronal network spontaneous electrical activity. Moreover, by means of the multiparametric analysis of neuronal network activity, the approach revealed a different toxicity mechanism of the cellular component compared to the algal conditioned growth medium, highlighting the potential active role of the first treatment.

Effects of the harmful dinoflagellate Ostreopsis cf. ovata on different life cycle stages of the common moon jellyfish Aurelia sp.

The frequency and geographic extension of microalgae and gelatinous zooplankton blooms seem to have been increasing worldwide over recent decades. In particular, the harmful dinoflagellate Ostreopsis cf. ovata and the Schyphozoan jellyfish Aurelia sp. are two of the most frequent and long lasting species forming blooms in the Mediterranean Sea. A kind of interaction among any of their life cycle stages (i.e. planula-polyp-ephyrae vs Ostreopsis cells) can likely occur, although in this area there are no data available on the co-occurrence of these species. The aim of this study was to investigate, for the first time, the potential noxious effect of O. cf. ovata on different life stages of Aurelia sp. (polyps and ephyrae), testing several concentrations of whole algal culture. Rsults of toxicity bioassay highlighted that ephyrae, but not polyps, are affected by this harmful dinoflagellate and comparisons among other model organisms show that Aurelia sp. ephyrae are the most sensitive model organism tested so far (EC50-24h=10.5cells/mL). These findings suggest an interesting scenario on the interaction of these two bloom forming species in the natural marine environment.

Chemical, molecular, and eco-toxicological investigation of Ostreopsis sp. from Cyprus Island: structural insights into four new ovatoxins by LC-HRMS/MS.

Blooms of benthic dinoflagellates of the genus Ostreopsis (mainly O. cf. ovata and occasionally O. cf. siamensis) represent a serious concern for humans in the Mediterranean area, due to production of palytoxin-like compounds listed among the most potent marine toxins known. In this work, six strains of Ostreopsis sp. from Cyprus Island were analyzed through an integrated approach based on molecular, chemical, and eco-toxicological methods. Cypriot Ostreopsis sp. was found to be a species distinct from O. cf. ovata and O. cf. siamensis, belonging to the Atlantic/Mediterranean Ostreopsis spp. clade. Some variability in toxin profiles emerged: three strains produced ovatoxin-a (OVTX-a), OVTX-d, OVTX-e, and isobaric palytoxin, so far found only in O. cf. ovata; the other three strains produced only new palytoxin-like compounds, which we named ovatoxin-i, ovatoxin-j1, ovatoxin-j2, and ovatoxin-k. The new ovatoxins present the same carbon skeleton as ovatoxin-a, differing primarily in an additional C2H2O2 moiety and an unsaturation in the region C49-C52. Other minor structural differences were found, including the presence of a hydroxyl group at C44 (in OVTX-j1 and OVTX-k) and the lack of a hydroxyl group in the region C53-C78 (in OVTX-i and OVTX-j1). The toxin content of the analyzed Ostreopsis sp. strains was in the range 0.06-2.8 pg cell(-1), definitely lower than that of a Ligurian O. cf. ovata strain cultured under the same conditions. Accordingly, an eco-toxicological test on Artemia salina nauplii demonstrated that Ostreopsis sp. presents a very low toxicity compared to O. cf. ovata. The whole of these data suggest that Ostreopsis sp. from Cyprus Island poses a relatively low risk to humans.