Cloning,Characterization, and Expression Levels of the <Emphasis Type="Italic">Nectin</Emphasis> Gene from the Tube Feet of the Sea Urchin <Emphasis Type="Italic">Paracentrotus Lividus</Emphasis>

Marine bioadhesives perform in ways that manmade products simply cannot match, especially in wet environments. Despite their technological potential, bioadhesive molecular mechanisms are still largely understudied, and sea urchin adhesion is no exception. These animals inhabit wave-swept shores, relying on specialized adhesive organs, tube feet, composed by an adhesive disc and a motile stem. The disc encloses a duo-gland adhesive system, producing adhesive and deadhesive secretions for strong reversible substratum attachment. The disclosure of sea urchin Paracentrotus lividus tube foot disc proteome led to the identification of a secreted adhesion protein, Nectin, never before reported in adult adhesive organs but, that given its adhesive function in eggs/embryos, was pointed out as a putative substratum adhesive protein in adults. To further understand Nectin involvement in sea urchin adhesion, Nectin cDNA was amplified for the first time from P. lividus adhesive organs, showing that not only the known Nectin mRNA, called Nectin-1 (GenBank AJ578435), is expressed in the adults tube feet but also a new mRNA sequence, called Nectin-2 (GenBank KT351732), differing in 15 missense nucleotide substitutions. Nectin genomic DNA was also obtained for the first time, indicating that both Nectin-1 and Nectin-2 derive from a single gene. In addition, expression analysis showed that both Nectins are overexpressed in tube feet discs, its expression being significantly higher in tube feet discs from sea urchins just after collection from the field relative to sea urchin from aquarium. These data further advocate for Nectin involvement in sea urchin reversible adhesion, suggesting that its expression might be regulated according to the hydrodynamic conditions.     

Do sulphuric acid and the brown alga <Emphasis Type="Italic">Desmarestia viridis</Emphasis> support community structure in Arctic kelp patches by altering grazing impact,distribution patterns,and behaviour of sea urchins?

Macrobenthic community structure and the distribution of the green sea urchin (Strongylocentrotus droebachiensis) were recorded inside and outside (=barrens) of kelp patches (Alaria esculenta) at Kongsfjordneset, Svalbard between August 2002 and October 2006. In manipulative field experiments, conducted at Kongsfjordneset, Svalbard in August 2002, the effect of the presence of the brown seaweed Desmarestia viridis on sea urchin distribution and kelp grazing was determined. Additionally, we studied the effect of sulphuric acid, which is produced and stored by D. viridis, on sea urchin movements in the laboratory at Ny-Ålesund, Svalbard, in May 2006. Sea urchin densities were two- to threefold lower in kelp patches than on barrens. The macrobenthic community inside kelp patches hosted 39% more species and was of different species composition than on barrens. Anchored pieces of the kelp A. esculenta were less consumed when surrounded by D. viridis than non-surrounded conspecifics. Changes in pH affected the behaviour of sea urchins. Exposing sea urchins to 500 μl seawater at pH 7.5 caused them to stop, while the exposure of as little as 25 μl at pH 1 caused sea urchins to move in the opposite direction. Acid-mediated escape responses in sea urchin behaviour suggest chemical protection by D. viridis as an additional mechanism to mechanical protection in the generation of kelp refuges. These results improve our understanding of how isolated kelp beds can persist over a wide range of environmental conditions, like wave-sheltered sites, and suggest that changes in community structure may be in part attributable to altered trophic interactions.     

Characterization of an Alpha Type Carbonic Anhydrase from <Emphasis Type="Italic">Paracentrotus lividus</Emphasis> Sea Urchin Embryos

Carbonic anhydrases (CA) are zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate. In the sea urchin, CA has a role in the formation of the calcitic skeleton during embryo development. Here, we report a newly identified mRNA sequence from embryos of the sea urchin Paracentrotus lividus, referred to as Pl-can. The complete coding sequence was identified with the aid of both EST databases and experimental procedures. Pl-CAN is a 447 aa-long protein, with an estimated molecular mass of 48.5 kDa and an isoelectric point of 6.83. The in silico study of functional domains showed, in addition to the alpha type CA-specific domain, the presence of an unexpected glycine-rich region at the N-terminal of the molecule. This is not found in any other species described so far, but probably it is restricted to the sea urchins. The phylogenetic analysis indicated that Pl-CAN is evolutionarily closer to human among chordates than to other species. The putative role(s) of the identified domains is discussed. The Pl-can temporal and spatial expression profiles, analyzed throughout embryo development by comparative qPCR and whole-mount in situ hybridization (WMISH), showed that Pl-can mRNA is specifically expressed in the primary mesenchyme cells (PMC) of the embryo and levels increase along with the growth of the embryonic skeleton, reaching a peak at the pluteus stage. A recombinant fusion protein was produced in E. coli and used to raise specific antibodies in mice recognized the endogenous Pl-CAN by Western blot in embryo extracts from gastrula and pluteus.     

Host location and selection by the symbiotic sargassum crab <Emphasis Type="Italic">Portunus sayi</Emphasis>: the role of chemical,visual and tactile cues

The Sargassum community consists of a unique and diverse assemblage of symbiotic fauna critical to pelagic food chains. Associated symbionts presumably have adaptations to assist in finding Sargassum. In situ scattered Sargassum patches accumulate as they are pushed toward the shoreline (via wind, waves, currents or tides) and are frequently less than 1 m apart and in depths of 10 cm or less as the patches approach the shoreline Crabs, and other symbiotic fauna, must relocate to another patch that is seaward in direction or likely perish as their current patch will likely become beached. This study investigated sensory cues used for host location and selection by the Sargassum crab, Portunus sayi. Chemical detection trials were conducted with a two-chamber choice apparatus with Sargassum spp. and Thalassia testudinum as habitat source odors. Visual detection trials (devoid of chemical cues) and habitat selection trials were conducted in which crabs were given a choice between hosts. Results showed that P. sayi responded to chemicals from Sargassum spp. Crabs visually located host habitats but did not visually distinguish between different hosts. In host selection trials, crabs selected Sargassum spp. over artificial Sargassum and T. testudinum. These results suggest that crabs isolated from Sargassum likely use chemoreception; within visual proximity of a potential patch, crabs likely use both chemical and visual information.     

Freezing tolerance of sea urchin embryo pigment cells

Various stresses, including exposure to cold or heat, can result in a sharp increase in pigmentation of sea urchin embryos and larvae. The differentiation of pigment cells is accompanied by active expression of genes involved in the biosynthesis of naphthoquinone pigments and appears to be a part of the defense system protecting sea urchins against harmful factors. To clarify numerous issues occurring at various time points after the cold injury, we studied the effect of shikimic acid, a precursor of naphthoquinone pigments, on cell viability and expression of some pigment genes such as the pks and sult before and after freezing the cultures of sea urchin embryo cells. The maximum level of the pks gene expression after a freezing–thawing cycle was found when sea urchin cells were frozen in the presence of trehalose alone. Despite naphthoquinone pigments have been reported to possess antioxidant and cryoprotectant properties, our data suggest that shikimic acid does not have any additional cryoprotective effect on freezing tolerance of sea urchin embryo pigment cells.     

Chemical composition of spines and tests of sea urchins

The skeleton of spines and tests of the species of sea urchins Strongylocentrotus intermedius, Mesocentrotus nudus, Scaphechinus mirabilis, and Echinocardium cordatum from the Sea of Japan is composed of a spongy stereom, consisting of calcite with a high content of magnesium. It was found that the tests and spines of the skeletons of sea urchins are composed of calcium–organic composite materials inlaid with other metals: Mg, Fe, Zn, and Rb. In the four species of sea urchins studied, the strength and other mechanical properties of the tests and spines differ and depend on the chemical composition and structural organization of their components. It was shown that the content of volatile substances correlates with their fragility or elasticity. It is revealed that the chemical composition of the tests of two species of the spherical sea urchins S. intermedius and M. nudus indicates significant differences between these two species of sea urchins.     

Non-native parasite enhances susceptibility of host to native predators

Parasites often alter host physiology and behavior, which can enhance predation risk for infected hosts. Higher consumption of parasitized prey can in turn lead to a less parasitized prey population (the healthy herd hypothesis). Loxothylacus panopaei is a non-native castrating barnacle parasite on the mud crab Eurypanopeus depressus along the Atlantic coast. Through prey choice mesocosm experiments and a field tethering experiment, we investigated whether the predatory crab Callinectes sapidus and other predators preferentially feed on E. depressus infected with L. panopaei. We found that C. sapidus preferentially consumed infected E. depressus 3 to 1 over visibly uninfected E. depressus in the mesocosm experiments. Similarly, infected E. depressus were consumed 1.2 to 1 over uninfected conspecifics in field tethering trials. We evaluated a mechanism behind this skewed prey choice, specifically whether L. panopaei affects E. depressus movement, making infected prey more vulnerable to predator attack. Counter to our expectations, infected E. depressus ran faster during laboratory trials than uninfected E. depressus, suggesting that quick movement may not decrease predation risk and seems instead to make the prey more vulnerable. Ultimately, the preferential consumption of L. panopaei-infected prey by C. sapidus highlights how interactions between organisms could affect where novel parasites are able to thrive.     

Percentage cover,biomass, distribution,and potential habitat mapping of natural macroalgae,based on high-resolution satellite data and in situ monitoring,at Libukang Island,Malasoro Bay,Indonesia

In this study, we combined remote sensing data and in situ observations to explore the potential habitats of macroalgae at Libukang Island, Indonesia. High-resolution satellite images from the GeoEye-1 were used to estimate and to map the geomorphological structures together with macroalgal species in the study area. Seasonal variations of percentage cover and biomass of macroalgae associated with substrates were investigated in May and November 2014, and June 2015, using quadrats as sampling unit. A total of nine common genera were found in the study area with three dominant genera: Sargassum, Padina, and Turbinaria. Most of macroalgae was observed in the eastern part of the Island, on several substrate types and particular oceanographic conditions (wave and current). Mean biomasses of Sargassum and Padina were high in May (1189.6 ± 455 and 166.7 ± 15.4 g DW.m^?2, respectively), while the biomass of Turbinaria was high in November (3245 ± 599.8 g DW.m^?2). The map accuracy of image classification for all typology substrates was 74.19%. Overall, approximately 62.3% of the total study area can be considered as potential for natural macroalgae habitats. Spectral response characteristic of shallow water substrates at study area based on GeoEye-1 is also presented. The results of this study exhibit a potential utilization of natural macroalgae in the study area, and provide information for a possible diversification of the use of macroalgae in Indonesia. The method could be useful for habitat management and future biomonitoring in the study area or other similar areas in Indonesia.     

<Emphasis Type="Italic">Diadema antillarum</Emphasis> 17 years after mass mortality: is recovery beginning on St. Croix?

Diadema antillarum was once ubiquitous in the Caribbean, but mass mortality in 1983–84 reduced its numbers by >97%. We measured Diadema abundance on back reefs and patch reefs that have been well studied for >25 years. From June 2000 to June 2001, populations on back reefs have increased >100% (June 2001 mean densities 0.004–0.368/m²), while patch reef populations increased >350% (June 2001 densities 0.236–0.516/m²). Populations are dominated by small urchins, suggesting high recent recruitment. Increased Diadema densities appear to be affecting macroalgae abundance. The general spatio-temporal pattern of recovery around St. Croix seems to be following that of the die-off, suggesting that the same oceanographic features that spread Diadema's pathogen are now carrying urchin larvae.     

The action of <Emphasis Type="Italic">Laminaria japonica</Emphasis> extractives on 1,3-β-D-glucanase,a digestive enzyme of the <Emphasis Type="Italic">Strongylocentrotus intermedius</Emphasis> sea urchin

Nutritional attractiveness of the brown alga Laminaria japonica for the sea urchin Strongylocentrotus intermedius was studied. The composition of L. japonica was analyzed after one and two years of its life under natural conditions, in its seedlings, and in the alga partially degraded by natural factors. Substances extracted with various solvents were tested for the presence of inhibitors and activators of 1,3-β-D-glucanase, a digestive enzyme of the sea urchin. Ethanolic extract of freshly harvested L. japonica was found to suppress the enzyme activity. Substances present in ethanolic extracts of the alga after one or two years of its life cycle and in the alga, partly degraded by natural factors, activated the sea urchin enzyme. This fact is in agreement with earlier natural observations concerning the nutritional attractiveness of such L. japonica samples for Strongylocentrotus intermedius.     

Nodal and BMP expression during the transition to pentamery in the sea urchin <Emphasis Type="Italic">Heliocidaris erythrogramma</Emphasis>: insights into patterning the enigmatic echinoderm body plan

Background

The molecular mechanisms underlying the development of the unusual echinoderm pentameral body plan and their likeness to mechanisms underlying the development of the bilateral plans of other deuterostomes are of interest in tracing body plan evolution. In this first study of the spatial expression of genes associated with Nodal and BMP2/4 signalling during the transition to pentamery in sea urchins, we investigate Heliocidaris erythrogramma, a species that provides access to the developing adult rudiment within days of fertilization.

Results

BMP2/4, and the putative downstream genes, Six1/2, Eya, Tbx2/3 and Msx were expressed in the earliest morphological manifestation of pentamery during development, the five hydrocoele lobes. The formation of the vestibular ectoderm, the specialized region overlying the left coelom that forms adult ectoderm, involved the expression of putative Nodal target genes Chordin, Gsc and BMP2/4 and putative BMP2/4 target genes Dlx, Msx and Tbx. The expression of Nodal, Lefty and Pitx2 in the right ectoderm, and Pitx2 in the right coelom, was as previously observed in other sea urchins.

Conclusion

That genes associated with Nodal and BMP2/4 signalling are expressed in the hydrocoele lobes, indicates that they have a role in the developmental transition to pentamery, contributing to our understanding of how the most unusual body plan in the Bilateria may have evolved. We suggest that the Nodal and BMP2/4 signalling cascades might have been duplicated or split during the evolution to pentamery.

     

Cilia are required for asymmetric <Emphasis Type="Italic">nodal</Emphasis> induction in the sea urchin embryo

Background

Left-right (LR) organ asymmetries are a common feature of metazoan animals. In many cases, laterality is established by a conserved asymmetric Nodal signaling cascade during embryogenesis. In most vertebrates, asymmetric nodal induction results from a cilia-driven leftward fluid flow at the left-right organizer (LRO), a ciliated epithelium present during gastrula/neurula stages. Conservation of LRO and flow beyond the vertebrates has not been reported yet.

Results

Here we study sea urchin embryos, which use nodal to establish larval LR asymmetry as well. Cilia were found in the archenteron of embryos undergoing gastrulation. Expression of foxj1 and dnah9 suggested that archenteron cilia were motile. Cilia were polarized to the posterior pole of cells, a prerequisite of directed flow. High-speed videography revealed rotating cilia in the archenteron slightly before asymmetric nodal induction. Removal of cilia through brief high salt treatments resulted in aberrant patterns of nodal expression. Our data demonstrate that cilia - like in vertebrates - are required for asymmetric nodal induction in sea urchin embryos.

Conclusions

Based on these results we argue that the anterior archenteron represents a bona fide LRO and propose that cilia-based symmetry breakage is a synapomorphy of the deuterostomes.

     

The distribution of macrobenthic epifauna in the Far Eastern Marine Reserve based on remote underwater video data

The spatial distribution of epifauna in the Far Eastern Marine Reserve was examined using a remotely operated vehicle. The abundance and distribution patterns of the sea stars Patiria pectinifera, Distolasterias nipon, and Asterias amurensis, as well as those of brittle stars, the black sea cucumber Cucumaria japonica, the sea urchin Strongylocentrotus intermedius, the ascidian Halocynthia aurantium, hermit crabs, and the Japanese scallop Mizuhopecten yessoensis were determined in the South Section of the Reserve. The average biomass of five dominant epifaunal species ranged from 51.0 to 87.4% of the average biomass of the softbottom communities.     

Mannan-binding lectins in the coelomic fluid of various species of Far Eastern echinoderms

A mannan-binding lectin activity was revealed in the coelomic fluid of the following echinoderm species inhabiting the coastal areas of the Sea of Japan, the holothurian Eupentacta fraudatrio, sea urchins Echinocardium cordatum, Strongylocentrotus nudus and S. intermedius, brittle star Amphipholis kochii, sea stars Asterina pectinifera, Lethasterias fusca, Lysastrosoma anthosticta, and Distolasterias nipon. It was shown that, concurrently with the general pattern of lectin interaction with branched bacterial mannans, there were also distinctions caused by the fine carbohydrate specificity of lectins. The obtained data preconditioned the further study of physical and chemical properties and structural features of the echinoderm MBL and the revelation of their role in the formation of the adaptive immune response and in other biological processes.     

Trophic Cascades Induced by Lobster Fishing Are Not Ubiquitous in Southern California Kelp Forests

Fishing can trigger trophic cascades that alter community structure and dynamics and thus modify ecosystem attributes. We combined ecological data of sea urchin and macroalgal abundance with fishery data of spiny lobster (Panulirus interruptus) landings to evaluate whether: (1) patterns in the abundance and biomass among lobster (predator), sea urchins (grazer), and macroalgae (primary producer) in giant kelp forest communities indicated the presence of top-down control on urchins and macroalgae, and (2) lobster fishing triggers a trophic cascade leading to increased sea urchin densities and decreased macroalgal biomass. Eight years of data from eight rocky subtidal reefs known to support giant kelp forests near Santa Barbara, CA, USA, were analyzed in three-tiered least-squares regression models to evaluate the relationships between: (1) lobster abundance and sea urchin density, and (2) sea urchin density and macroalgal biomass. The models included reef physical structure and water depth. Results revealed a trend towards decreasing urchin density with increasing lobster abundance but little evidence that urchins control the biomass of macroalgae. Urchin density was highly correlated with habitat structure, although not water depth. To evaluate whether fishing triggered a trophic cascade we pooled data across all treatments to examine the extent to which sea urchin density and macroalgal biomass were related to the intensity of lobster fishing (as indicated by the density of traps pulled). We found that, with one exception, sea urchins remained more abundant at heavily fished sites, supporting the idea that fishing for lobsters releases top-down control on urchin grazers. Macroalgal biomass, however, was positively correlated with lobster fishing intensity, which contradicts the trophic cascade model. Collectively, our results suggest that factors other than urchin grazing play a major role in controlling macroalgal biomass in southern California kelp forests, and that lobster fishing does not always catalyze a top-down trophic cascade.     

Predation of <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Hemiptera: Pentatomidae) from Web-Building Spiders Associated with Anthropogenic Dwellings

The brown marmorated stink bug, or Halyomorpha halys, is an invasive pest in North America and Europe that causes severe agricultural damage and nuisance problems for homeowners; and it is originally from China, Taiwan, and the Republic of Korea. While the natural enemy community of H. halys has been evaluated in several agroecosystems, it has not been examined where H. halys overwinters in anthropogenic structures. The aims of the current study were to evaluate 1) whether spider webs commonly found in the home and yard can successfully ensnare H. halys, 2) whether entanglement resulted in consumption by spiders inhabiting the webs, and 3) how frequently H. halys becomes entangled in webs under ambient conditions. To accomplish this, adult H. halys were introduced into webs in and near anthropogenic structures in West Virginia and Maryland, United States, and the behavior of spiders was observed for 5-min intervals at 0, 1, 2, and 24 h after introduction. In addition, a survey of webs was performed to determine the frequency with which spiders naturally capture H. halys inside buildings and in the landscape. Overall, the study found seven spider families in anthropogenic structures. Adult H. halys that were introduced into the webs of Theridiidae, Pholcidae, or Agelenidae had a greater than 50% chance of being ensnared and consumed. Adult H. halys were found naturally most often in webs of Theridiidae. Webs with a funnel or cob web architecture had the greatest probability of ensnaring H. halys, while those with orb structures resulted in the fewest caught. In the wild, 13–20% of spider webs contained dead H. halys. Our results suggest that spiders may be an important contributing factor for mortality of H. halys at overwintering sites, and spiders in or outside homes may help reduce nuisance problems caused by H. halys.     

Distribution and ecological relations among the alien crab, <Emphasis Type="Italic">Percnon gibbesi</Emphasis> (H. Milne-Edwards 1853) and autochthonous species,in and out of an SW Mediterranean MPA

The distribution of the allochthonous crab Percnon gibbesi and its relationships with other benthic invertebrate species was assessed inside the marine reserve of Cabo de Palos—Islas Hormigas (Mediterranean sea, Spain) and neighbouring non-protected sites. Although a significant spatial variability was detected at finer spatial scale, there was no influence of protection measures or insularity on the abundance of P. gibbesi. The presence of small holes, encrusting algae and low slope favour the colonization success of this crustacean, and the spatial distribution of these habitat features could explain the observed pattern. The abundance of P. gibbesi was similar to that of native crab species; however, a non-significant negative relationship between the abundance of P. gibbesi and native crabs (Pachygrapsus marmoratus and Eriphia verrucosa), urchins (Arbacia lixula and Paracentrotus lividus) and a snail (Phorcus turbinatus) was observed. This work highlights the importance of monitoring alien crab population densities taking structural habitat and other potentially influential factors into account and the likely effect of this alien species on the native ones.     

Genetic diversity of free-living <Emphasis Type="Italic">Symbiodinium</Emphasis> in the Caribbean: the importance of habitats and seasons

Although reef corals are dependent of the dinoflagellate Symbiodinium, the large majority of corals spawn gametes that do not contain their vital symbiont. This suggests the existence of a pool of Symbiodinium in the environment, of which surprisingly little is known. Reefs around Curaçao (Caribbean) were sampled for free-living Symbiodinium at three time periods (summer 2009, summer 2010, and winter 2010) to characterize different habitats (water column, coral rubble, sediment, the macroalgae Halimeda spp., Dictyota spp., and Lobophora variegata, and the seagrass Thalassia testudinum) that could serve as environmental sources of symbionts for corals. We detected the common clades of Symbiodinium that engage in symbiosis with Caribbean coral hosts A, B, and C using Symbiodinium-specific primers of the hypervariable region of the chloroplast 23S ribosomal DNA gene. We also discovered clade G and, for the first time in the Caribbean, the presence of free-living Symbiodinium clades F and H. Additionally, this study expands the habitat range of free-living Symbiodinium as environmental Symbiodinium was detected in T. testudinum seagrass beds. The patterns of association between free-living Symbiodinium types and habitats were shown to be complex. An interesting, strong association was seen between some clade A sequence types and sediment, suggesting that sediment could be a niche where clade A radiated from a free-living ancestor. Other interesting relationships were seen between sequence types of Symbiodinium clade C with Halimeda spp. and clades B and F with T. testudinium. These relationships highlight the importance of some macroalgae and seagrasses in hosting free-living Symbiodinium. Finally, studies spanning beyond a 1-yr cycle are needed to further expand on our results in order to better understand the variation of Symbiodinium in the environment through time. All together, results presented here showed that the great diversity of free-living Symbiodinium has a dynamic distribution across habitats and time.     

Intertidal benthic communities associated with the macroalgae <Emphasis Type="Italic">Iridaea cordata</Emphasis> and <Emphasis Type="Italic">Adenocystis utricularis</Emphasis> in King George Island,Antarctica

Antarctic benthos has been a main target in Antarctic research, but very few quantitative studies have been carried out in the littoral zone, which may be seasonally covered by macroalgae. In this work, we studied (1) cover and biomass of the macroalgae Iridaea cordata and Adenocystis utricularis, and (2) composition of macrobenthic assemblage associated with these macroalgal species at three locations at King George Island: Mareograph Beach (1 M), Tank’s Bay (2R) and Ardley Bay (3R). Iridaea cordata was collected by completely detaching the algae from the substrate, while A. utricularis was scraped. Adenocystis utricularis covered more than 80 % of the substrate at all locations, while coverage of Iridaea cordata was below 53 % or absent (3R). Fresh biomass of I. cordata was 0.8–61.4 g/individual and 4.7–93.0 g/100 cm² for A. utricularis. The assemblage associated with both macroalgae differed significantly between sites. The studied fauna was composed mainly of amphipods, gastropods and bivalves. Species diversity was higher in the community associated with A. utricularis. A total of ~27 ind/g DW were found associated with I. cordata, while ~112 ind/g DW were found associated with A. utricularis. The most abundant groups associated with I. cordata were amphipods at 1 M (57 %) and gastropods at 2R (46 %). Both groups were responsible for the dissimilarity between localities (62.50 %). The most abundant groups associated with A. utricularis were the gastropods at all localities reaching up to 82 % at 1 M. This study provides a first baseline on the diversity and abundance of benthic assemblages associated with intertidal macroalgae in the southwest of King George Island.