WntA and Wnt4 during the regeneration of internal organs in the holothurian Eupentacta fraudatrix.

BACKGROUND: Over the past few years, it has been established that wnt genes are involved in the regenerative processes of holothurians. The wnt4 gene was identified as one of the most active genes in Eupentacta fraudatrix regeneration using differential gene expression analysis and qPCR of individual genes. Also, the wntA gene was found in holothurians, which is present only in invertebrates and can perform unique functions. RESULTS: In this regard, both these genes and proteins were studied in this work. During regeneration, the Wnt4 protein is found in the cells of the coelomic and ambulacral epithelium, retractor muscles, and radial nerves. Single cells with this protein are also found in the connective tissue of the developing aquapharyngeal bulb and in the hypoderm of the body wall. Cells with WntA are found exclusively in the hypoderm of the body wall. CONCLUSION: We assume that both genes are involved in regeneration, but Wnt4 coordinates the formation of the epithelial tissue structure, while WntA maintains the state of the intercellular substance of the body wall.

Role of smoothened and sfrp genes in Eupentacta fraudatrix regeneration.

The secreted frizzled-related proteins (sfrp) and smoothened (smo) genes and their possible role in the regeneration of internal organs in the holothurian Eupentacta fraudatrix were studied. In this species, two sfrp genes were identified: sfrp1/2/5, sfrp3/4 and one smo gene. Their expression was analysed during regeneration of the aquapharyngeal bulb (AB) and intestine, and these genes were knock down by RNA interference. It has been shown that the expression of these genes is extremely important for the formation of AB. In all animals subjected to knockdown, at 7 days after evisceration, a full-sized AB rudiment was not formed. As a result of sfrp1/2/5 knockdown, the process of extracellular matrix remodelling in AB is interrupted, that leading to clusters of dense connective tissue formation, which slows down cell migration. When sfrp3/4 is knockdown, the connective tissue of AB anlage is completely disrupted and its symmetry is broken. The effect of smo knockdown was expressed in a significant impairment of AB regeneration, when connections between ambulacras were not formed after evisceration. However, despite severe disturbances in AB regeneration, a normal-sized gut anlage developed in all cases, which suggests that the regeneration of the digestive tube and AB occur independently of each other.

Sex-specific seasonal variations in the fatty acid and carotenoid composition of sea cucumber gonads and implications for aquaculture.

Fatty acids and carotenoids are known to have roles in embryonic and larval development of sea cucumbers, but their changes in gonads during gametogenesis have not yet been studied. To improve our knowledge of the reproductive cycle of sea cucumbers in an aquaculture perspective, we collected 6-11 individuals of the species Holothuria (Panningothuria) forskali Delle Chiaje, 1823 approximately every 2 months from December 2019 to July 2021 east of the Glenan Islands (Brittany - France; 47.710°N, 3.948°W) at a depth of 8-12 m. Our results show that soon after spawning, sea cucumbers take advantage of an increased food availability in spring to rapidly and opportunistically accumulate nutrients in the form of lipids in their gonads (from May to July) and then slowly elongate, desaturate and probably rearrange fatty acids within lipid classes for the next reproductive season according to the specific requirements of both sexes. In contrast, acquisition of carotenoids occurs synchronously with gonads filling and/or through the reabsorption of spent tubules (T5), thus revealing little seasonal variations at the scale of the entire gonad in terms of relative abundance in both sexes. All results suggest that gonads are fully replenished with nutrients by October and that broodstock for induced reproduction could be captured at this moment and kept until the production of larvae is required. Maintaining broodstock for consecutive years would probably be a higher level challenge as the dynamics of tubule recruitment are not fully understood and seem to last for several years. Supplementary Information: The online version contains supplementary material available at 10.1007/s00227-023-04198-0.

How Biological Activity in Sea Cucumbers Changes as a Function of Species and Tissue.

Biological activity and bioactive compound content in sea cucumbers was assessed, considering Parastichopus regalis, Holothuria mammata, Holothuria forskali, and Holothuria arguinensis as species and intestine, muscle band, respiratory tree, body wall, and gonads as tissues. P. regalis had the lowest content in phenolic compounds and antioxidant activity in contrast to Holothuria species. In the respiratory tree, the highest phenolic concentration was recorded in H. arguinensis, 76.4 ± 1.2 mg GAE/100 g dw vs. 21.0-49.0 mg GAE/100 g dw in the other species. H. arguinensis had the highest DPPH and FRAP results in the gonads, 13.6 ± 0.7 mg AAE/100 g dw vs. 2.6-3.5 mg AAE/100 g dw and 27.1 ± 0.3 µmol Fe2+/g dw vs. 8.0-15.9 µmol Fe2+/g dw, respectively. Overall, P. regalis biomass presented the highest anti-inflammatory activity levels and H. arguinensis the lowest anti-inflammatory levels. The respiratory tree was the most anti-inflammatory (measured by the inhibition of cyclooxygenase-2, COX-2) tissue in H. mammata and H. forskali (also the muscle band in this case), 76.3 ± 6.3% and 59.5 ± 3.6% COX-2 inhibition in 1 mg/mL aqueous extracts, respectively. The results demonstrated a variable bioactive potential and advantage in targeting antioxidant properties in the muscle band and anti-inflammatory activity in the respiratory tree, which may constitute a starting point for a biorefinery approach envisaging multiple applications.

Muscle Regeneration in Holothurians without the Upregulation of Muscle Genes.

The holothurian Eupentacta fraudatrix is capable of fully restoring its muscles after transverse dissection. Although the regeneration of these structures is well studied at the cellular level, the molecular basis of the process remains poorly understood. To identify genes that may be involved in the regulation of muscle regeneration, the transcriptome of the longitudinal muscle band of E. fraudatrix has been sequenced at different time periods post-injury. An analysis of the map of biological processes and pathways has shown that most genes associated with myogenesis decrease their expression during the regeneration. The only exception is the genes united by the GO term "heart valve development". This may indicate the antiquity of mechanisms of mesodermal structure transformation, which was co-opted into various morphogeneses in deuterostomes. Two groups of genes that play a key role in the regeneration have been analyzed: transcription factors and matrix metalloproteinases. A total of six transcription factor genes (Ef-HOX5, Ef-ZEB2, Ef-RARB, Ef-RUNX1, Ef-SOX17, and Ef-ZNF318) and seven matrix metalloproteinase genes (Ef-MMP11, Ef-MMP13, Ef-MMP13-1, Ef-MMP16-2, Ef-MMP16-3, Ef-MMP24, and Ef-MMP24-1) showing differential expression during myogenesis have been revealed. The identified genes are assumed to be involved in the muscle regeneration in holothurians.

Shallow epifaunal sea cucumber densities and their relationship with the benthic community in the Okinawa Islands.

Sea cucumbers are important ecological engineers in marine ecosystems. However, the fishery demand of some species, especially large-epifaunal and commercially used (LEC) sea cucumbers, has risen drastically, resulting in serious depletion of local populations for many species. Despite this problem, basic ecological data on sea cucumbers, such as population densities and preferred habitats, are often still insufficient. Here, we report on the population densities of multiple LEC sea cucumber species, and their ambient benthic communities at eight sites around Okinawa Islands. Further, we discuss the correspondence between sea cucumber densities and the surrounding coral communities. Our results show two sites within national or quasi-national parks, Aka and Manza, where stricter rules have been placed on fisheries and land reclamation compared to other areas, had the highest and third highest sea cucumber population densities among sites, respectively. Holothuria atra was observed at all survey sites and made up the majority of sea cucumber populations at all sites except for Chatan and Sesoko, where Holothuria leucospilota and Stichopus chloronotus were most abundant, respectively. Regarding the relationships between benthic composition and LEC sea cucumber species, S. chloronotus was significantly correlated with dead corals, scleractinian corals, and coralline algae. As well, H. leucospilota had significant correlations with rubble. Although there were no significant correlations between any specific scleractinian coral genus and sea cucumber densities, S. chloronotus was marginally insignificant with Platygyra and Psammocora. Notably, medium- to highly valued species were sparse in our surveys, and most of them appeared at only one site. Additionally, at one site (Odo), only three LEC sea cucumber individuals were observed. Combining these facts with relatively low population densities around the Okinawa Islands compared to densities reported in previous research from the Indo-West Pacific Ocean region, we conclude that Okinawan LEC sea cucumber populations have been and are being impacted by high levels of direct (e.g., overexploitation, as well as coastal development) and indirect anthropogenic pressure (e.g., decreasing water quality). To address the current situation, repeated monitoring and more detailed investigations to reveal the drivers that determine LEC sea cucumber species aggregations and population densities are urgently needed, along with more robust management of remaining LEC sea cucumber populations.

Characterization and Expression Analysis of Regeneration-Associated Protein (Aj-Orpin) during Intestinal Regeneration in the Sea Cucumber Apostichopus japonicus.

Apostichopus japonicus achieves intestinal regeneration in a short period after evisceration, and multiple genes are involved in this process. The transcriptome of A. japonicus was screened for regeneration-associated protein (Aj-Orpin), a gene that is specifically upregulated during intestinal regeneration. The expression and function of Aj-Orpin were identified and investigated in this study. The 5' and 3' RACE polymerase chain reaction (PCR) was used to clone the full-length cDNA of Aj-Orpin. The open reading frame codes for a 164 amino-acid protein with an EF-hand_7 domain and overlapping signal peptides and transmembrane regions. Moreover, Aj-Orpin mRNA and protein expression during intestinal regeneration was investigated using real-time quantitative PCR and Western blot. The expression pattern of Aj-Orpin in the regenerating intestine was investigated using immunohistochemistry. The results showed that Aj-Orpin is an exocrine protein with two EF-hand-like calcium-binding domains. Expression levels were higher in the regenerating intestine than in the normal intestine, but protein expression changes lagged behind mRNA expression changes. Aj-Orpin was found to play a role in the formation of blastema and lumen. It was primarily expressed in the serosal layer and submucosa, suggesting that it might be involved in proliferation. These observations lay the foundation for understanding the role of Orpin-like in echinoderm intestinal regeneration.

Effects of Human Activity on Markers of Oxidative Stress in the Intestine of Holothuria tubulosa, with Special Reference to the Presence of Microplastics.

Pollution in the seas and oceans is a global problem, which highlights emerging pollutants and plastics, specifically microplastics (MPs), which are tiny (1 µm to 5 mm) ubiquitous plastic particles present in marine environments that can be ingested by a wide range of organisms. Holothurians are benthic organisms that feed on sediment; therefore, they can be exposed to contaminants present in the particles they ingest. The objective was to evaluate the effects of human activity on Holothuria tubulosa through the study of biomarkers. Specimens were collected in three different areas throughout the island of Eivissa, Spain: (1) a highly urbanized area, with tourist uses and a marina; (2) an urbanized area close to the mouth of a torrent; (3) an area devoid of human activity and considered clean. The results showed a higher presence of microplastics (MPs) in the sediments from the highly urbanized area in relation to the other two areas studied. Similarly, a higher number of MPs were observed in the digestive tract of H. tubulosa from the most affected area, decreasing with the degree of anthropic influence. Both in the sediment and in the holothurians, fibers predominated with more than 75% of the items. In the three areas, mesoplastics were analyzed by means of FTIR, showing that the main polymer was polypropylene (27%) followed by low-density polyethylene (17%) and polystyrene (16%). Regarding the biomarkers of oxidative stress, the intestine of H. tubulosa from the most impacted areas showed higher catalase, superoxide dismutase (SOD), glutathione reductase (GRd), and glutathione S-transferase (GST) activities and reduced glutathione (GSH) levels compared to the control area. The intermediate area only presented significant differences in GRd and GST with respect to the clean area. The activities of acetylcholinesterase and the levels and malondialdehyde presented similar values in all areas. In conclusion, human activity evaluated with the presence of MPs induced an antioxidant response in H. tubulosa, although without evidence of oxidative damage or neurotoxicity. H. tubulosa, due to its benthic animal characteristics and easy handling, can be a useful species for monitoring purposes.

New psychropotid species (Echinodermata, Holothuroidea, Elasipodida) of the Western Pacific with phylogenetic analyses.

Holothurians of the family Psychropotidae are widely distributed but remain the least studied deep-sea holothurians. On an expedition to the Western Pacific, six psychropotid specimens were collected by the Jiaolong Human Operated Vehicle (HOV). Through morphological examination, four of them were identified as a new species, Benthodytesjiaolongi sp. nov., which was characterized as having minute papillae, a narrow brim, and a terminal anus; and the ossicles were rods and primary crosses. The remaining two specimens were identified as Psychropotesverrucicaudatus Xiao, Gong, Kou & Li, 2019, first recorded at the Kyushu-Palau Ridge. The phylogenetic analysis showed that B.jiaolongi sp. nov. and P.verrucicaudatus were embedded in the clades Benthodytes and Psycheotrephes, respectively, and that Benthodytes was paraphyletic. The new species clustered with Benthodytessanguinolenta and was separated from the clade containing the other Benthodytes species.

Quorum Quenching Strains Isolated from the Microbiota of Sea Anemones and Holothurians Attenuate Vibriocorallilyticus Virulence Factors and Reduce Mortality in Artemiasalina.

Interference with quorum-sensing (QS) intercellular communication systems by the enzymatic disruption of N-acylhomoserine lactones (AHLs) in Gram-negative bacteria has become a promising strategy to fight bacterial infections. In this study, seven strains previously isolated from marine invertebrates and selected for their ability to degrade C6 and C10-HSL, were identified as Acinetobacter junii, Ruegeria atlantica, Microbulbifer echini, Reinheimera aquimaris, and Pseudomonas sihuiensis. AHL-degrading activity against a wide range of synthetic AHLs were identified by using an agar well diffusion assay and Agrobacterium tumefaciens NTL4 and Chromobacterium violaceum CV026 and VIR07 as biosensors. High-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis indicated that this activity was not due to an AHL lactonase. All the strains degraded Vibrio coralliilyticus AHLs in coculture experiments, while some strains reduced or abolished the production of virulence factors. In vivo assays showed that strains M3-111 and M3-127 reduced this pathogen's virulence and increased the survival rate of Artemia salina up to 3-fold, indicating its potential use for biotechnological purposes. To our knowledge, this is the first study to describe AHL-degrading activities in some of these marine species. These findings highlight that the microbiota associated with marine invertebrates constitute an important underexplored source of biological valuable compounds.

Sophistication in a seemingly simple creature: a review of wild holothurian nutrition in marine ecosystems.

Holothurians are marine invertebrates that are among the most widespread benthic megafauna communities by both biomass and abundance in shallow-water and deep-sea ecosystems, their functions supporting important ecological services worldwide. Despite their simple appearance as sea cucumbers, holothurians show a wide range of feeding practices. However, information on what and how these animals eat is scattered and potentially confusing. We provide a comprehensive review of holothurian nutrition in coastal and deep-sea ecosystems. First, we describe morphological aspects of holothurian feeding and the ultrastructure of tentacles. We discuss the two processes for food capture, concluding that mucus adhesion is likely the main method; two mucous cells, type-1 and type-2, possibly allow the adhesion and de-adhesion, respectively, of food particles. Secondly, this review aims to clarify behavioural aspects of holothurian suspension- and deposit-feeding. We discuss the daily feeding cycle, and selective feeding strategies. We conclude that there is selectivity for fine and organically rich particles, and that feeding through the cloaca is also a route for nutrient absorption. Third, we provide a wide description of the diet of holothurians, which can be split into two categories: living and non-living material. We suggest that Synallactida, Molpadida, Persiculida, Holothuriida and Elasipodida, ingest the same fractions, and emphasise the importance of bacteria in the diet of holothurians.

WITHDRAWN: Sea cucumbers of the Arabian Peninsula and Iran - A review of historical and current research trends.

Sea cucumbers are benthic marine invertebrates with immense ecological and commercial value. Processed sea cucumbers known as "Beche-de-mer" are a delicacy in southeast Asian countries with an ever-increasing demand depleting wild stocks on a global scale. Aquaculture techniques are well developed for commercially important species (eg. Holothuria scabra) to aid in conservation and trade. In the Arabian Peninsula and Iran, where the major land mass is surrounded by marginal seas (Arabian/Persian Gulf, Gulf of Oman, Arabian Sea, Gulf of Aden, and Red Sea), studies on sea cucumbers are rather limited and its economic value is underestimated. Historical and current research trends indicate impoverished diversity (82 species) due to environmental extremes. Artisanal fisheries exist for the sea cucumbers of Iran, Oman, and Saudi Arabia, with Yemen and United Arab Emirates (UAE) playing a key role in collection and export to Asian countries. Stock assessment and data on export indicates depletion of natural stocks in Saudi Arabia and Oman. Aquaculture trials of high value species (H. scabra) were successful in Saudi Arabia, Oman and Iran with prospects for further expansion. Research on ecotoxicological properties and bioactive substances conducted in Iran demonstrates an immense research potential. Molecular phylogeny, biology, use in bioremediation, and characterisation of bioactive compounds were identified as potential gaps in research. Expanding aquaculture operations could revive exports and recuperate damaged stocks through sea ranching. Furthermore, regional cooperation, networking, training, and capacity building could help fill the gaps in sea cucumber research, which will aid in its effective conservation and management.

Current Progress in Lipidomics of Marine Invertebrates.

Marine invertebrates are a paraphyletic group that comprises more than 90% of all marine animal species. Lipids form the structural basis of cell membranes, are utilized as an energy reserve by all marine invertebrates, and are, therefore, considered important indicators of their ecology and biochemistry. The nutritional value of commercial invertebrates directly depends on their lipid composition. The lipid classes and fatty acids of marine invertebrates have been studied in detail, but data on their lipidomes (the profiles of all lipid molecules) remain very limited. To date, lipidomes or their parts are known only for a few species of mollusks, coral polyps, ascidians, jellyfish, sea anemones, sponges, sea stars, sea urchins, sea cucumbers, crabs, copepods, shrimp, and squid. This paper reviews various features of the lipid molecular species of these animals. The results of the application of the lipidomic approach in ecology, embryology, physiology, lipid biosynthesis, and in studies on the nutritional value of marine invertebrates are also discussed. The possible applications of lipidomics in the study of marine invertebrates are considered.

Sea cucumbers of the Arabian Peninsula and Iran - A review of historical and current research trends.

Sea cucumbers are benthic marine invertebrates with immense ecological and commercial value. Processed sea cucumbers known as "Beche-de-mer" are a delicacy in southeast Asian countries with an ever-increasing demand depleting wild stocks on a global scale. Aquaculture techniques are well developed for commercially important species (e.g. Holothuria scabra) to aid in conservation and trade. In the Arabian Peninsula and Iran, where the major land mass is surrounded by marginal seas (Arabian Gulf, Gulf of Oman, Arabian Sea, Gulf of Aden, and Red Sea), studies on sea cucumbers are rather limited and its economic value is underestimated. Historical and current research trends indicate impoverished diversity (82 species) due to environmental extremes. Artisanal fisheries exist for the sea cucumbers of Iran, Oman, and Saudi Arabia, with Yemen and United Arab Emirates (UAE) playing a key role in collection and export to Asian countries. Stock assessment and data on export indicates depletion of natural stocks in Saudi Arabia and Oman. Aquaculture trials of high value species (H. scabra) were successful in Saudi Arabia, Oman and Iran with prospects for further expansion. Research on ecotoxicological properties and bioactive substances conducted in Iran demonstrates an immense research potential. Molecular phylogeny, biology, use in bioremediation, and characterisation of bioactive compounds were identified as potential gaps in research. Expanding aquaculture operations could revive exports and recuperate damaged stocks through sea ranching. Furthermore, regional cooperation, networking, training, and capacity building could help fill the gaps in sea cucumber research, which will aid in its effective conservation and management.

The sea cucumber Holothuria tubulosa does not reduce the size of microplastics but enhances their resuspension in the water column.

Microplastic pollution is increasingly recognized as a prominent threat to marine life. Understanding the role of bioturbators is crucial to determine to what extent marine sediments can act as a microplastic sink. The presence of microplastics has been documented in holothurians, but no study has investigated how the ingestion-egestion process influences their bioavailability. Using the Mediterranean deposit-feeder, Holothuria tubulosa, as a model system, we assessed if, upon ingestion, plastic particles are accumulated in pseudofeces and if the passage through the digestive tract reduces their size. To this end, the number, shape and colour of plastic particles was compared between pseudofeces and surrounding surficial sediments collected along the edges of a seagrass meadow. Pseudofeces were enriched in plastic fragments with respect to surficial sediments, suggesting a selective ingestion of fragments over fibres. By contrast, there was no difference in the size or colour of plastic particles between pseudofeces and sediments. In addition, by means of a laboratory experiment, we evaluated how microplastic resuspension rates from pseudofeces compares with those from surficial sediments. Under standard water movement conditions, the resuspension of labelled microplastics from pseudofeces was much greater than that from sediments (i.e., about 92% and 26% at the end of the experimental trial). Greater relative abundance of fine material (i.e., pelite) in pseudofeces than sediments could explain their physical instability and, hence, their lower microplastic retention. Our results suggest that pseudofeces of H. tubulosa not only represent a hotspot for plastic fragment concentration, but, due to their surficial deposition and rapid dissolution, they could also promote their transfer to the water column. Ingestion and egestion of microplastics by this sea cucumber, although not altering their size, may thus enhance their bioavailability.

Molecular Aspects of Regeneration Mechanisms in Holothurians.

Holothurians, or sea cucumbers, belong to the phylum Echinodermata. They show good regenerative abilities. The present review provides an analysis of available data on the molecular aspects of regeneration mechanisms in holothurians. The genes and signaling pathways activated during the asexual reproduction and the formation of the anterior and posterior parts of the body, as well as the molecular mechanisms that provide regeneration of the nervous and digestive systems, are considered here. Damage causes a strong stress response, the signs of which are recorded even at late regeneration stages. In holothurian tissues, the concentrations of reactive oxygen species and antioxidant enzymes increase. Furthermore, the cellular and humoral components of the immune system are activated. Extracellular matrix remodeling and Wnt signaling play a major role in the regeneration in holothurians. All available morphological and molecular data show that the dedifferentiation of specialized cells in the remnant of the organ and the epithelial morphogenesis constitute the basis of regeneration in holothurians. However, depending on the type of damage, the mechanisms of regeneration may differ significantly in the spatial organization of regeneration process, the involvement of different cell types, and the depth of reprogramming of their genome (dedifferentiation or transdifferentiation).

Assessment of Megadenus holothuricola Rosén, 1910 (Eulimidae), an endoparasite of Holothuria mexicana Ludwig, 1875 (Holothuriidae) in the southern Gulf of Mexico and the description a new species.

As part of a study on holothurians from the southern Gulf of Mexico, some Holothuria mexicana Ludwig, 1875 were obtained for gut analysis. In two of them, a couple of eulimids were located inside the main tube of the respiratory tree. They were identified as Megadenus holothuricola Rosén, 1910, described from the Bahamas Islands, based on five specimens attached to the respiratory tree of H. mexicana. The original description was brief with few details, the type material is lost, and the species has not been found again. In this contribution, this species is confirmed for Campeche Bay, Mexico. The adult shell is globular to conical, transparent, thin, and fragile. Megadenus smithi sp. nov. from Palmyra Atoll, Central Pacific is described based on adult specimens. It differs from its congeneric species in its more robust shell, the pseudopallium does not cover the shell, and its short and contracted proboscis forms a thick disc. Further research on these eulimid parasites is now complicated in the southern Gulf of Mexico because of the holothurian population collapse due to over-exploitation of the fishery.

Assessment of the impacts of glyphosate and its commercial formulation Roundup® on the respiratory tree of the sea cucumber Holothuria forskali using a multivariate biomarker approach.

In this study, the potential hazardous impacts of the technical grade glyphosate acid (GLY) and its commercial formulation roundup (RD®) were evaluated for the first time on holothurians. To do this, redox status, fatty acid (FA) profile, and histopathology aspects were assessed in the respiratory tree tissue of the sea cucumber Holothuria forskali following short-term exposure (96 h) to a series of concentrations (10, 100 and 1000 µg L-1) of GLY and RD® (glyphosate acid equivalent). Our results showed that both GLY and RD® promoted oxidative stress highlighted with an increase of hydrogen peroxide (H2O2), malondialdehyde (MDA), lipid peroxides (LOOH) and advanced oxidation protein product (AOPP) levels in all treated groups. In addition, both glyphosate forms were found to perturb the FA composition. However, changes in saturated (SFA) and polyunsaturated (PUFA) including some essential FA (LA, ARA, EPA and DHA) revealed differential compensatory/adaptive processes in H. forskali depending on the treatment. GLY and RD® were also found to modulate the enzymatic (glutathione S-transferases, glutathione peroxidase and catalase) and non-enzymatic (reduced glutathione and ascorbic acid) antioxidant defense status. Taken together, our results revealed that the commercial formulation produced more pronounced effects on H. forskali respiratory tree than the pure form. This finding was further confirmed by the histological observations.

Two new species of holothurians of the genus Echinopsolus Gutt, 1990(Echinodermata: Dendrochirotida: Cucumariidae) from the North-Western Pacific.

Two new species of the genus Echinopsolus-E. sanamyanorum and E. onekotanensis are described. This genus is new for North-Western Pacific marine fauna. Echinopsolus sanamyanorum was collected in the Avacha Gulf (south-east coast of Kamchatka), E. onekotanensis-near the Onekotan Island (The Kuril Islands).

Chemical composition of microplastic in sediments and protected detritivores from different marine habitats (Salina Island).

This study estimates chemical composition of microplastic in sediments and benthic detritivores (sea cucumbers) collected from different marine rocky bottom habitat types (bank, landslide, cliff) of Salina Island (Aeolian Archipelago, Italy). Also, species richness and bottom coverings by benthic species were recorded at each sampling station. Correlations among chemical composition of microplastic in sediments and in detritivores were explored linking recorded variability to the factor "habitat type". Results evidence that the habitat types considered in this study are characterized by wide species richness and by high percentages of bottom coverage by protected species by international conventions. In spite of the high ecological value of habitats considered in this study, microplastics were recorded both in sediments (PVC, PET, PE, PS, PA, PP) and in stomach contents of sea cucumbers (PET, PA) collected in all sampling sites, confirming the exposure of benthic species to microplastic pollution.