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Quinones are one of the major pigment groups that provide such bright colors to feather stars (Echinodermata, Crinoidea). These secondary metabolites also act as defensive molecules rendering crinoids unpalatable and repellent to other organisms. However, feather stars are usually associated with numerous symbiotic organisms, amongst which the ectocommensal snapping shrimp Synalpheus stimpsonii. We investigated the chemical stimulus allowing host selection in S. stimpsonii through the combination of behavioral tests, chemical extractions, and mass spectrometry analyses. The individuals of S. stimpsonii used in the experiments were sampled around the Great Reef of Toliara (Madagascar) where they are found in association with two crinoid species: Comanthus wahlbergii and Phanogenia distincta. The chemical attractiveness of the two crinoid hosts and a non-host species, Cenometra bella, was tested in an olfactometer. The three crinoids produced attractive kairomones allowing the snapping shrimp to recognize them. Mass spectrometry analyses on purified extracts of P. distincta revealed the presence of three different anthraquinones (rhodoptilometrin, comantherin, and a new crinoid anthraquinone). Compared to the existing literature, this anthraquinonic cocktail is specific to P. distincta. When these extracts were injected in the olfactometer, they triggered similar attracting behavior suggesting that crinoid anthraquinones are kairomones allowing host selection for S. stimpsonii. This hypothesis is also supported by the fact that shrimps were chemically attracted by pure commercial anthraquinones. In addition to their traditional defensive role (allomones), anthraquinones would, therefore, also function as kairomones, maintaining the symbiosis between S. stimpsonii and its crinoid hosts.

International audience; 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 suspensionand 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.

The distribution of larvae and recruitment of scleractinians in the southwest region of Madagascar were evaluated for the first time between October 2013 and September 2014 at 3 sites. The presence of coral larvae (planulae) was monitored through weekly sampling using a plankton net and the recruitment rate evaluated by monthly sampling of the newly settled corals ( 10 juveniles m-2) compared to other regions and the threshold, but it revealed high mortality among recruits. Coral recruitment in the southwest region of Madagascar was found to be high and could result in increased resilience of the coral reef assemblages.

Holothuria scabra is one of the most intensively studied holothuroids, or sea cucumbers (Echinodermata: Holothuroidea), having been discussed in the literature since the early 19th century. The species is important for several reasons: (1) it is widely distributed and historically abundant in several shallow soft-bottom habitats throughout the Indo-Pacific, (2) it has a high commercial value on the Asian markets, where it is mainly sold as a dried product (beche-de-mer) and (3) it is the only tropical holothuroid species that can currently be mass-produced in hatcheries. Over 20 years have elapsed since the last comprehensive review on H. scabra published in 2001. Research on H. scabra has continued to accumulate, fuelled by intense commercial exploitation, and further declines in wild stocks over the entire distribution range. This review compiles data from over 950 publications pertaining to the biology, ecology, physiology, biochemical composition, aquaculture, fishery, processing and trade of H. scabra, presenting the most complete synthesis to date, including scientific papers and material published by local institutions and/or in foreign languages. The main goal of this project was to summarize and critically discuss the abundant literature on this species, making it more readily accessible to all stakeholders aiming to conduct fundamental and applied research on H. scabra, or wishing to develop aquaculture, stock enhancement and management programs across its geographic range.

Due to the dearth of molecular markers variable enough to distinguish species of black corals, species delimitation in Antipatharia is still mainly based on morphological traits. One of the few markers available, the internal transcribed spacer (ITS) region located in ribosomal DNA, has already been used several times to infer phylogenetic relationships among antipatharians. However, this region was never considered in population-scale studies of shallow-water black corals till now. Here, we sampled two phenotypic variants of 41 specimens of the shallow-water antipatharian Stichopathes cf. maldivensis along the SW of Madagascar. The sampling area represents 200 km of shore and encompasses the runoffs of three rivers. We used direct sequencing to determine the main ITS1 types present in the genomes of the collected specimens and retrieved sequences from GenBank for Indonesian specimens. Then, we constructed a haploweb to visualize possible species boundaries using the criterion of mutual allelic exclusivity. With this method, we confirm that the previously described phenotypic variants belong to a single species that apparently extends from Indonesia to Madagascar. Likewise, the three rivers that pour freshwater in southwestern Madagascar do not appear to act as a barrier for the larval dispersal of this species. This study highlights the usefulness of ITS markers within antipatharians at both species and population levels.

Aquacultivated sea cucumbers often suffer from SKin Ulceration Diseases (SKUDs). SKUDs have been observed in six holothuroid species from nine countries. All SKUDs present a similar symptom—the skin ulceration—and can be induced by bacteria, viruses, or abiotic factors. We here provide an update on SKUDs in holothuroids and analyse the case of the SKUD observed in Holothuria scabra in Madagascar. Field observations revealed a seasonality of the disease (i.e. wintertime maximum peak). Morphological analyses of integument ulcers showed that sea cucumbers react by forming a collagen fibre plug. Metagenomic analyses revealed a higher proportion of Vibrionaceae (Gammaproteobacteria) in ulcers in comparison to the healthy integument of the same individuals. Experimental infection assays were performed with ulcer crude extracts and bacteria isolated from these extracts (e.g. Vibrio parahaemolyticus) but did not significantly induce skin ulceration. Our results suggest that the disease is not induced by a pathogen or, at the very least, that the pathogen is not found within the ulcers as the disease is not transmissible by contact. An initial cause of the SKUD in Madagascar might be the repeated and prolonged exposures to cold temperatures. Opportunistic bacteria could settle in the dermis of ulcerated individuals and promote the ulcer extension. We propose a general nomenclature for SKUDs based on the acronym of the disease, the affected sea cucumber species (e.g. Hs for Holothuria scabra), the concerned region using an ISO code 3166-2 (e.g. MG for Madagascar), the description date (e.g. 20 for the year 2020), and, when known, the inducing agent (first letter of the general taxon, b for bacteria, v for virus in currently known cases; a a if it is an abiotic inducing parameter; nothing if the inducing cause has not been precisely identified). The disease described in this work will be designated under the name SKUD Hs-MG-20.

Crustaceans are associated with a wide diversity of hosts, using both chemical and visual cues to recognize them. The pea crab Dissodactylus primitivus is an ectoparasite of two species of irregular echinoids living in the Caribbean Sea. Previous studies showed that the crab chemically discriminates its hosts from non-host species. The possibility that the parasite also visually localizes its host was investigated here through behavioral and morphological approaches. The responses of the parasite to visual cues were investigated in aquaria and show a limited visual ability, leading to sheltering rather than to host localization. This suggests that visual cues are not required to maintain the specificity of the parasitism. Microscopical investigations corroborate this conclusion by revealing a pair of small compound eyes mainly localized under the cephalothorax. The ommatidia (facets) were only found on the covered surface (below the cuticle). Interestingly, a lot of setae were observed around or even directly on the eye and might participate in the overall chemical detection.

In holothuroids, oocyte maturation is stopped in ovaries at the prophase I stage of meiosis. In natural conditions, the blockage is removed during the spawning by an unknown mechanism. When oocytes are isolated by dissection, the meiotic release can be successfully induced by a natural inducer, the REES (i.e., Rough Extract of Echinoid Spawn) that is used in aquaculture to obtain viable larvae in mass. A thioredoxin has recently been identified in the REES as the molecule responsible for holothuroid oocyte maturation. As a redox-active protein, thioredoxin is thought to reduce target proteins within the oocyte membrane and initiate an intracellular reaction cascade that leads to the unblocking of the oocyte meiosis. Our results allow us to understand additional steps in the intracellular reaction cascade induced by the action of thioredoxin on oocytes. Pharmacological agents known to have activating or inhibiting actions on oocyte maturation have been used (Forskolin, Isobutylmethylxanthine, Hypoxanthine, 6-dimethyaminopurine, Lavendustin, Genistein, Roscovitine, Cycloheximide). The effects of these agents were analysed on oocytes of the holothuroid Holothuria tubulosa incubated with or without REES and were compared to those obtained with another reducing agent, the dithiothreitol. Our results demonstrated that, at the opposite of dithiothreitol-induced oocyte maturation, thioredoxin-induced oocyte maturation is cAMP independent, but dependent of the presence of calcium in the seawater. Both pathways of induction require the activation of protein serine/threonine kinases.

Random sampling of shallow-water black corals colonies of Cirrhipathes anguina was carried out over a period of 13 months near the Great Reef of Toliara in southwest Madagascar. Small piece...

Chromatographic fractions of a rough extract of echinoid spawn (REES) is demonstrated to efficiently induce oocyte maturation in aspidochirote holothuroids. The method is so efficient that it is currently used in holothuriculture to obtain fertilised eggs even outside the reproductive period of the aquacultivated species. In the present study, we isolated and identified from echinoid spawns the molecule responsible for the induction of the oocyte maturation in the Mediterranean holothuroid Holothuria tubulosa and the Indo-Pacific Holothuria scabra. The use of proteinase K and dialysing membranes suggests that the active molecule is a protein with a molecular weight superior to 12,000 kDa. The active molecule has been isolated on G-100 Sephadex chromatography column. Active chromatographic fractions include seven proteins identified with nanoLC-MS/MS technique. One of them, identified as a thioredoxin-2 and to which the name Trx-REES was given, gave levels of maturation similar to those obtained with the REES and with a commercial thioredoxin extracted from Escherichia coli. Occurrence of thioredoxin in REES was confirmed by immunoblot analysis, and the maturation-inducing properties of thioredoxin were positively checked using anti-thioredoxin antibodies. A peptide of 6 AAs corresponding to the active site of Trx-REES, composed of WCNPCK, was synthesised and its efficiency in holothuroid oocyte maturation was tested. At some concentrations, the isolated peptide was 1.2 times more active than the REES. This short peptide may be used as an effective tool for artificial induction of oocyte maturation in the sea cucumber aquaculture context.

Tuleariocaris holthuisi and Arete indicus are two ectocommensal shrimps closely associated with the tropical sea urchin Echinometra mathaei. This study provides a comparison of these two E. mathaei symbiotic crustaceans and particularly focuses on the relationship between T. holthuisi and its host’s pigments (i.e. spinochromes), and its dependency on its host. While all the analyses underline a close association between A. indicus and E. mathaei, they reveal a particularly close interaction between T. holthuisi and its host. Chemical analyses reveal that these shrimps present the same spinochrome composition as E. mathaei, and have similar colouration, allowing camouflage. Isotopic composition and pigment loss after host separation suggest that these pigments are certainly assimilated upon feeding on the urchin. Moreover, symbiont isolation experiments demonstrate the high dependency of T. holthuisi on its host and the importance of the host’s pigments on their survival capacity. Finally, some host recognition mechanisms are investigated for T. holthuisi and show the probable implication of spinochromes in host selection, through chemical recognition. Hence, all the results suggest the essential roles of spinochromes for T. holthuisi, which, in turn, suggests the potential implication of these pigments in the shrimps’ metabolism.

In non-model organisms, Next Generation Sequencing (NGS) technology improve our ability to analyze gene expression and identify new genes or transcripts of interest. In this research, paired-end Illumina HiSeq sequencing has been used to describe a composite transcriptome based on two libraries generated from dorsal and ventral integuments of the European sea cucumber Holothuria forskali (Holothuroidea, Echinodermata). A total of 43,044,977 million HQ reads were initially generated. After de novo assembly, a total of 111,194 unigenes were predicted. On all predicted unigenes, 32,569 show significant matches with genes/proteins present in the reference databases. Around 50% of annotated unigenes were significantly similar to sequences from the purple sea urchin Strongylocentrotus purpuratus genome. Annotation analyses were performed on predicted unigenes using public reference databases. These RNA-seq data provide an interesting resource for researchers with a broad interest in sea cucumber biology.

Here, we report the first methodological approach to investigate the presence and estimate the density of Symbiodiniaceae cells in corals of the order Antipatharia subclass Hexacorallia, known as black corals. Antipatharians are understudied ecosystem engineers of shallow (200 m) reefs. They provide habitat to a vast number of marine fauna, enhancing and supporting coral reefs biodiversity globally. Nonetheless, little biological and ecological information exists on antipatharians, including the extent at which global change disturbances are threatening these corals. The assumption that they were exempted from threats related to climate change was challenged by findings of high density of dinoflagellates within three antipatharian colonies. Further methodical studies were necessary to investigate the regularity of these findings. An integrated design combining microscopy and molecular techniques was used to investigate the presence and estimate density of Symbiodiniaceae cells within two antipatharians species -Cupressopathes abies and Stichopathes maldivensis -from shallow and mesophotic reefs of SW Madagascar. Symbiodiniaceae-like cells were found within the two species from both shallow and mesophotic reefs, although the overall cell density was very low (0-4 cell mm-3). These findings suggest that high abundance of Symbiodiniaceae is not characteristic of antipatharians, which has relevant implications considering disruptions associated to climate change affecting other corals. However, the high densities of dinoflagellates found in antipatharian colonies exposed to higher light irradiance in other studies should be further examined.

We provide the first record of the shallow and mesophotic (Antipathes grandis VERRILL, 1928 from the Indian Ocean. First described from Hawaii, A. grandis was recently found on a mesophotic coral reef in SW Madagascar. Its identity was confirmed by means of morphological and molecular analyses (cytochrome c oxidase subunit I). These findings not only expand our knowledge of the potential geographic range of antipatharian corals, but also challenge the belief that only deep-sea (> 200 m depth) benthic taxa have such wide geographic distributions.

Antipatharians, also known as black corals, represent a small group of anthozoan hexacorallians found in all oceans of the world. They are generally considered a deep-water taxon; however, some of the most diverse communities are known from tropical shallow waters. With a few poorly detailed exceptions, shallow-water black corals from the Indian Ocean and especially those from Madagascar are mostly unknown. In this study, we report for the first time a highly diverse black coral assemblage of the Western Indian Ocean thriving in shallow waters and upper mesophotic depths (10–52 m depth) along the SW coast of Madagascar. A total of 22 species belonging to six genera (Antipathes, Arachnopathes, Cirrhipathes, Cupressopathes, Myriopathes and Stichopathes) and two families (Antipathidae and Myriopathidae) are described, of which 20 are found in the northern pass of the Great Reef of Toliara, thus representing the most diverse site of the areas investigated. Most of the shallow-water species from the Indian Ocean were originally described more than a century ago, sometimes without being reported again until now. All the descriptions herein rely solely on morphology and include detailed in situ pictures and scanning electron microscope images, in addition to range expansions for many species.

Saponins are plant secondary metabolites. There are associated with defensive roles due to their cytotoxicity and are active against microorganisms. Saponins are frequently targeted to develop efficient drugs. Plant biomass containing saponins deserves sustained interest to develop high-added value applications. A key issue when considering the use of saponins for human healthcare is their toxicity that must be modulated before envisaging any biomedical application. This can only go through understanding the saponin-membrane interactions. Quinoa is abundantly consumed worldwide, but the quinoa husk is discarded due to its astringent taste associated with its saponin content. Here, we focus on the saponins of the quinoa husk extract (QE). We qualitatively and quantitively characterized the QE saponins using mass spectrometry. They are bidesmosidic molecules, with two oligosaccharidic chains appended on the aglycone with two different linkages, a glycosidic bond and an ester function. The latter can be hydrolyzed to prepare monodesmosidic molecules. The microwave-assisted hydrolysis reaction was optimized to produce monodesmosidic saponins. The membranolytic activity of the saponins was assayed based on their hemolytic activity that was shown to be drastically increased upon hydrolysis. In silico investigations confirmed that the monodesmosidic saponins interact preferentially with a model phospholipid bilayer, explaining the measured increased hemolytic activity.

In order to test the feasibility of transplantation of the whip black coral species Cirrhipathes anguina (Dana, 1846) from Madagascar, transplants were installed on cultivation tables in two sites (the North Pass and the Grande Vasque) characterized by distinct environmental conditions. Following transplantation, the transplants were followed for short-term (20 days) healing capacities and medium-term (200 days) mortality and growth rates. Results show a successful transplantation in both sites with 0% mortality (except in the form of missing fragments) and a certain growth over 200 days. Maximum growth rates were 3.4 cm/month and 2.0 cm/month in the North Pass and in the Grande Vasque, respectively. In addition, mean time to total healing was delayed in the Grande Vasque compared to the transplants in the North Pass (10 days). Differences between sites are likely to be related to differences in environmental parameters. Altogether, the promising results obtained with the easy protocol used in this study encourage the use of black coral transplants in further in situ experiments and restoration projects.

Amphiura filiformis is a common burrowing brittle star from muddy environments along the European shores. This species emits a blue light at the level of the arm spine tips when mechanically stimulated. Although the putative luciferase of A. filiformis was recently discovered, the ultrastructure of the luminous cells, i.e. the photocytes, remain unknown for this species as well as for most other bioluminescent echinoderms. In this study, we describe the morphology of the arm and spines in the brittle star A. filiformis and compare it to the luciferase expression pattern revealed by immunohistochemistry. Luciferase is expressed at the base of the spine and at the arm tips within well-defined photocyte clusters. Additionally, we investigated the ultrastructure of spine tissues before and after chemical stimulation of light emission in order to identify photocytes and improve our comprehension of the photogenesis phenomenon in brittle stars. Type II granular cells, found in the basal to mid-basal portion of the spine inner tissues and showing ultrastructural modifications during light-emission, are proposed as photocytes. Putative photocytes appear intimately associated with the spine nerve plexus as well as with mucus cells and presumptive pigment cells. Photocytes are characterised by the presence of specific vesicles with reticulated contents in their cytoplasm. Furthermore, the structure of the calcified spine ossicle has been investigated using microCT scanning.

Since a couple of decades, coral aquaculture has been developed in many countries to face an increasing live coral market and to support conservation of endangered natural coral reefs. To evaluate the potentiality of community-based coral aquaculture in Madagascar, we experienced suitable farming techniques using the species Acropora nasuta and Seriatopora caliendrum . Survival and growth rate of the nubbins were monitored during wet, warm and dry, cold seasons. To determine economical feasibility, the coral market was investigated and the yields were also calculated using the technical, biological and social parameters of the production. Coral nubbins were reared in situ at appropriate conditions. Coral nubbins reared during the wet, warm season showed a final survival rate of 67 ± 6% and 57 ± 4% respectively for A. nasuta and S. caliendrum , while in the dry, cold season, the survival rates were of 85 ± 7% and 69 ± 1% respectively. A. nasuta had a significantly higher survival rate than S. caliendrum during both seasons. During the wet, warm season, growth rates were 0.46 ± 0.16% d − 1 and 0.54 ± 0.16% d − 1 respectively for A. nasuta and S. caliendrum . In the dry, cold season, A. nasuta had 0.63 ± 0.18% d − 1 of growth rate, while S. caliendrum grew 0.65 ± 0.15% d − 1 . Significant difference was observed between both species during the wet, warm season, but not during the dry, cold season. Furthermore, both species grew faster during the dry, cold season. These results are in the range of reference values for corals. The activity can be profitable from 25 coral nubbins sold per month. Profit can already be perceived from the second year and a total of more than EUR 27,000 earned after 5 years of developing project, for an initial investment of EUR 1978. Marine animals wholesale companies and biodiversity conservation NGOs seem to be the appropriate clients for this form of aquaculture on Madagascar. Statement of relevance This paper provides new form of coral aquaculture: the community-based coral aquaculture. The socially and environmentally responsible production of coral is among the benefits of this new economically viable form of aquaculture.

Echinoderms form a remarkable phylum of marine invertebrates that present specific chemical signatures unique in the animal kingdom. It is particularly the case for essential triterpenoids that evolved separately in each of the five echinoderm classes. Indeed, while most animals have Δ5-sterols, sea cucumbers (Holothuroidea) and sea stars (Asteroidea) also possess Δ7 and Δ9(11)-sterols, a characteristic not shared with brittle stars (Ophiuroidea), sea urchins (Echinoidea), and crinoids (Crinoidea). These particular Δ7 and Δ9(11) sterols emerged as a self-protection against membranolytic saponins that only sea cucumbers and sea stars produce as a defense mechanism. The diversity of saponins is large; several hundred molecules have been described in the two classes of these saponins (i.e., triterpenoid or steroid saponins). This review aims to highlight the diversity of triterpenoids in echinoderms by focusing on sterols and triterpenoid glycosides, but more importantly to provide an updated view of the biosynthesis of these molecules in echinoderms.

The scleractinian biodiversity of Madagascar is mainly known from one study performed in the Bay of Toliara (SW of Madagascar) in the 1970s. In the present study, this biodiversity was re-investigated 40 years later, at 2 sites previously considered as atypical, but now subject to high anthropogenic pressures. Results showed lower species diversity compared to the previous study, and to similar sites in the Indian Ocean region, but most of the well-rep- resented genera were recorded. The occurrence of previously unrecorded species suggests that the scleractinian communities are changing, in addition to declining. The findings of the present study constitute a baseline of scle - ractinian structure studies, focused on diversity change. Further investigations on this reef must consider these changes, and management measures must be adapted to ensure greater efficiency.

Holothuria (Platyperona) sanctori is a common sea cucumber of the Mediterranean Sea. Although its ecology and molecular phylogeny have been investigated in previous works, nothing is known about its saponin content, contrarily to other Mediterranean holothuroids. Saponins are bioactive triterpene glycosides that present interesting pharmacological/biological properties, and which can be used as taxonomic markers. This work highlights the different saponins produced by H. sanctori . Mass spectrometry ( MALDI-MS and MS/MS ) analyses highlighted a total of 18 different saponins, including eight new congeners. Presumptive molecular structures are proposed for all these molecules. A higher diversity of saponins is found in the body wall (12) than in the Cuvierian tubules (8). In addition to this qualitative difference, saponins were also 2–3 fold more concentrated in the Cuvierian tubules than in the integument. Our results demonstrate that H . sanctori , in accordance with its phylogenetic position, is the second species of Holothuria described to date possessing exclusively non-sulfated saponins. The species presents its own mix of saponins, acting as a true chemical signature.

This study describes the association between the obligatory symbiont coral gall crab Hapalocarcinus marsupialis and its stony coral hosts Seriatopora sp. within the Great Reef of Toliara in Madagascar and attempts to discuss their symbiotic status through comparison with previous studies. These corals are inhabited by crabs living in galls that can be categorised in four distinct morphological stages, where the first one corresponds to a small bud and the last one represents a completely closed gall surrounding the crab inside. Within the reef, 563 colonies of Seriatopora species were observed by scuba-diving at ten different stations: 37.8 % of them were infested by H. marsupialis, with a total of 763 galls, and with a majority of stage 4 galls. Galls are monopolised by females that can have different morphologies. Females store the sperm in two spermathecae and are fertilised when their morphology and size are similar to males and the gall is not closed. Histological observations coupled with scanning electronic microscopy analyses show that closed galls are made of an external living tissue, a mid skeletal layer and an internal living tissue. The internal living tissue includes polyps similar to the external tissue, some of them being sexually mature. Nitrogen and carbon isotopic signatures confirmed that these crabs are filter-feeders and do not feed on their host. This association perfectly highlights the difficulties to define the symbiotic status of a symbiont if one considers inflexible the three categories of symbiosis commonly defined.

With the increase of seaweed farming activities, epiphytic filamentous algae (EFA) disease has appeared in many regions of Madagascar. This infestation has dramatic consequences for local farmers as it alters drastically farmed algal growth and has caused farming activity to collapse in many places. The present study characterizes the structure and ultrastructure of the stages observed in the life cycle of Polysiphonia sp. and gives the results of a monitoring of 18 months made in three Kappaphycus alvarezii farming sites in the southwest of Madagascar. Transmission electron microscopy (TEM) was used to analyze the ultrastructure of the cortex in infested K. alvarezii. Five stages have been observed in the life cycle of Polysiphonia sp.: the infesting stage that is a small dark spot observed at the surface of K. alvarezii, the male gametophyte, the female gametophyte, the tetrasporocysts, and the undifferentiated stage where individuals show normal thalli without sexual differentiation. EFA infestation was never recorded in Sarodrano, but often in the two other monitored villages (Lambohara, Tampolove). Prevalence of infestation varied from 40 to 100 % and the rates of infestation from 42 to 78 epiphytes cm−2. Prevalence of infestation showed significant seasonal variation and a between-sites variation; the rates of infestation were not significantly different between sites and did not vary with the period. The ways of infestation between K. alvarezii individuals in an infested field and from infested to healthy fields are discussed at the light of the present results.

Algae biomass is nowadays considered as a unique reservoir of a huge variety of different molecules, such as lipids, carbohydrates, proteins and secondary metabolites, awaiting development in various fields. In particular, the valorization of the algae polysaccharide content is still in its infancy and intense research activity is starting to appear. Oligosaccharides with defined molecular weight and structures are extremely interesting for different applications such as plant biostimulation or biopesticide. Oligosaccharide are often prepared in medium to large scale upon chemical-based hydrolysis reactions from the extracted polysaccharides in harsh conditions (temperature and pH), leading to the loss of the fragile groups such as the sulfate functions in carrageenans. In the present work, microwave-based hydrolysis, as a green chemistry method, is optimized to produce bioactive oligosaccharides from carrageenans. Microwave hydrolysis process is first optimized on a commercially available carrageenan and further applied on carrageenans extracted from two algae, namely Kappaphycus alvarezii or Eucheuma spinosum that are produced in large quantities in the world and have a high content of carrageenans. We also develop a complete set of characterization methods, i.e. size exclusion chromatography, reducing sugar and residual sulfate determination and mass spectrometry, to perform the follow-up of the hydrolysis reaction. We here demonstrate that fast (5 min) and controlled hydrolysis of carrageenans can be achieved in pure water under microwave activation, while preserving the nature of the active sulfate groups (>95%).

Chromatographic fractions of a rough extract of echinoid spawn (REES) has been demonstrated to efficiently induce oocyte maturation in aspidochirote holothuroids. The method is so efficient that it is now used in holothuriculture to get fertilised eggs even outside the reproductive period of the aquacultured species. We here isolate and identify from echinoid spawns the molecule responsible for the induction of the oocyte maturation in the Mediterranean holothuroidHolothuria tubulosaand the Indo-PacificHolothuria scabra. The use of proteinase K and dialysing membranes indicates that the active molecule is a protein with a molecular weight superior to 12,000 kDa. The active molecule has been isolated on G-100 Sephadex chromatography column. Active chromatographic fractions include seven proteins identified with nanoLC-MS/MS technique. One of them, identify as a thioredoxin-2 and to which the nameTrx-REEShas been given, leads up to levels of maturation similar to those obtained with the REES and with a commercial thioredoxin extracted fromEscherichia coli. Occurrence of thioredoxin in REES was confirmed by immunoblot analysis, and the maturation-inducing properties of thioredoxin were positively checked in using anti-thioredoxin antibodies. A peptide of 6 AAs corresponding to the active site ofTrx-REES, composed of WCNPCK, was synthesised and its efficiency in holothuroid oocyte maturation tested. At some concentrations, the peptide was 1.2 times more active than the REES.

Sea urchin pigmentation is mainly due to polyhydroxy-1,4-naphthoquinones called spinochromes. If their molecular structures are well known in test and spines of many species, their abundance and distribution in other body compartments remain unstudied. The aim of this study is to analyse the pigment composition in four body compartments (test/spines, digestive system, gonads and coelomic fluid) of four coloured types of the sea urchin Echinometra mathaei . Qualitative and quantitative measurements by mass spectrometry highlight the existence of 13 different pigments; among which are five isomers of known spinochromes as well as three potentially new ones. The composition comparison shows the largest spinochrome diversity in ‘test/spines’ body compartments. The spinochrome concentrations vary from 48 to 1279 mg kg −1 of dried body compartment. It is the highest in the digestive system, although it is also important in the organic fraction of the ‘test/spines’ body compartment. This observation may be explained by higher exposures of some body compartments to external environments and by the protective role fulfilled by spinochromes against microorganisms, ultraviolet radiation and reactive oxygen species. The ‘black’ type—the most common coloured type in coral reefs—has the highest concentration of spinochromes indicating their importance in Echinoids' fitness by acting as a protective agent.

Mesomyzostoma Remscheid, 1918 currently includes three described species that live in the coelom and/or gonads of comatulid crinoids: Mesomyzostoma reichenspergeri Remscheid, 1918, Mesomyzostoma katoi Okada, 1933 and Mesomyzostoma lanterbecqae Summers and Rouse, 2014 in Summers, Al-Hakim et al. 2014. Here we describe four new species of Mesomyzostoma and assess their phylogenetic relationships using 18S rRNA, cytochrome oxidase subunit I and 16S rRNA sequence data. We also designate a neotype for M. katoi as the original types appear to be lost. We record M. reichenspergeri from the Australian Great Barrier Reef and from northern Papua New Guinea, but samples from the type locality (Aru Islands, Indonesia) and previously recorded host are needed for confirmation. The new species of Mesomyzostoma are one Japanese species: Mesomyzostoma okadai sp. nov., and three Australian species: Mesomyzostoma lobus sp. nov., Mesomyzostoma leukos sp. nov. and Mesomyzostoma botulus sp. nov. The first infects the c...

Sea urchin pigmentation is mainly due to polyhydroxy-1,4-naphthoquinones called spinochromes. If their molecular structures are well known in test and spines of many species, their abundance and distribution in other body compartments remain unstudied. The aim of this study is to analyse the pigment composition in four body compartments (test/spines, digestive system, gonads and coelomic fluid) of four coloured types of the sea urchin

Spinochromes are principally known to be involved in sea urchin pigmentation as well as for their potentially interesting pharmacological properties. To assess their biological role in sea urchin physiology, experiments are undertaken on crude extracts from four species and on four isolated spinochromes in order to test their antibacterial, antioxidant, inflammatory and cytotoxic activities. First, the antibacterial assays show that the use of crude extracts as representatives of antibacterial effects of spinochromes are inaccurate. The assays on purified spinochromes showed a decrease in the growth of four strains with an intensity depending on the spinochromes/bacteria system, revealing the participation of spinochromes in the defense system against microorganisms. Secondly, in the 2,2-diphenyl-1-picrylhydrazyl antioxidant assays, spinochromes show an enhanced activity compared to the positive control. This latter observation suggests their involvement in ultraviolet radiation protection. Third, spinochromes present a pro-inflammatory effect on lipopolysaccharide-stimulated macrophages, highlighting their possible implication in the sea urchin immune system. Finally, cytotoxicity assays based on Trypan blue exclusion, performed in view of their possible future applications as drugs, show a weak cytotoxicity of these compounds against human cells. In conclusion, all results confirm the implication of spinochromes in sea urchin defense mechanisms against their external environment and reveal their potential for pharmacological and agronomical industries.

High sedimentation rates have been linked to reduced coral health within multiple systems; however, whether this is a direct result of compromised coral immunity has not been previously investigated. The potential effects of sedimentation on immunity of the hard coral Montipora patula were examined by comparing physiological responses of coral fragments inoculated with sterilized marine sediments and those under control conditions. Sediments were collected from terrestrial runoff-affected reefs in SW Madagascar and applied cyclically for a total of 24 h at a rate observed during precipitation-induced sedimentation events. Coral health was determined 24 h after the onset of the sedimentation stress through measuring metabolic proxies of O2 budget and lipid ratios. Immune response of the melanin synthesis pathway was measured by quantifying phenoloxidase activity and melanin deposits. Sedimentation induced both immune and metabolic responses in M. patula. Both phenoloxidase activity and melanin deposition were significantly higher in the sediment treatment compared to controls, indicating an induced immune response. Sediment-treated corals also showed a tendency towards increased respiration (during the night) and decreased photosynthesis (during the day) and a significant depletion of energy reserves as compared to controls. These data highlight that short-term (24 h) sedimentation, free of live microorganisms, compromises the health of M. patula. The energetically costly immune response, potentially elicited by residual endotoxins and other inflammatory particles associated with the sterile sediments, likely contributes to the energy depletion. Overall, exposure to sedimentation adversely affects coral health and continued exposure may lead to resource depletion and an increased susceptibility to disease.

DespiteHolothuria scabra's wide distribution and status as one of the best candidates for sustaining the development of tropical sea cucumber aquaculture, very few data are available regarding the organic fraction it assimilates in practice. In this paper we report experimental results whereH. scabra's diet was supplemented with various15N-labelled organic fractions of sediment. We used juveniles weighing between 38 and 88 mg at the beginning of the experiment (~2 cm long and 30 days old). Their growth was measured over a four-week period and their15N composition recorded. The results showed thatH. scabrajuveniles assimilated all added organic components from both dissolved and particulate fractions of the sediment. Bacteria seem to be an important food source for juveniles, even more so than microphytobenthos (diatoms).

The present paper characterizes, for the first time, the diet of the Harlequin crab Lissocarcinus orbicularis, an obligate symbiotic crab that associates with sea cucumbers (holothuroids) belonging to the genera Thelenota, Bohadschia and Holothuria. These tropical holothuroids host a rich symbiotic community in the Indo-West Pacific Ocean of which the Harlequin crab is the best known. The diet of L. orbicularis was characterized by analyzing the microscopic, molecular and isotopic signatures obtained from its gastric content. The presence of sea cucumber ossicles in the gastric mills of the crabs suggests that symbionts eat the superficial integument of their host and this was supported by the fact that Holothuroid DNA was detected in the stomach of L. orbicularis after DGGE and sequencing of the 18S rDNA gene. The stable isotopic δ13C and δ15N values of crab tissues were compared with diverse potential food sources including three holothuroids, three algae, one sea grass as well as the organic matter contained in the water column, in the sediment, and the second most abundant symbiont, the polychaete Gastrolepidia clavigera. The low δ15N values of crabs suggests that the crabs do not exclusively feed on sea cucumber tissue but assimilate diverse food sources such as sea grasses and organic matter contained in sediment that have similar δ13C values. There were no differences between the feeding of males and females but there was a positive correlation between the carapace length and the stable isotopic values indicating a shift of the food source as crabs grow larger.