Apluteal development of the sea urchin Holopneustes purpurescens Agassiz (Echinodermata: Echinoidea: Euechinoidea)

The external features of a shortened, apluteal development (lacking a pluteus larva) are described. Some features are unusual for echinoids. The large egg is distinctively marked by dark and pale coloured yolk. The sperm entry point is marked by a dark yolk spot and the first cleavage plane in most embryos is through the meridian on which the sperm entry point lies. Dark yolk in the animal hemisphere segregates largely to one blastomere in the two-cell embryo and pale yolk segregates to the other as a result of yolk movements during the first cell cycle. Progeny of the pale-yolk blastomere form adult oral structures and progeny of the dark-yolk blastomere form adult aboral structures. There is no feeding planktonic pluteus larva. The gastrula develops into a demersal vestibula larva with bilateral symmetry. The plane of symmetry is coincident with the Carpenter axis that defines a plane of symmetry through the madreporite in adult echinoderms. The coincidence shows that the anterior ambulacrum is vegetal with respect to egg polarity and the interradius originating at the madreporite is animal. The bilateral symmetry of the vestibula offers insight into the origin of radial symmetry in echinoderms and the body plan of an echinoderm ancestor.     

Evolutionary history of larval skeletal morphology in sea urchin Echinometridae (Echinoidea: Echinodermata) as deduced from mitochondrial DNA molecular phylogeny

The larval skeletons of sea urchins show considerable morphological diversity, even between closely related species, although the evolutionary history and functional significance of this diversity are poorly understood. To infer the evolutionary history of the skeletal morphology, we focused on echinometrid species for which the morphological variation in larval skeletons had been investigated qualitatively and quantitatively. We reconstructed the phylogenetic relationships among 14 echinometrid species based on mitochondrial ND1 and ND2 genes and mapped the morphological characters onto the resultant trees. The monophyly of each genus in the Echinometridae was well supported by our results, as was the close affinity between Colobocentrotus, Heterocentrotus, and Echinometra. The mapping of the morphological characters of the larval skeletons indicated that the length, direction, and density of spines on the postoral rods was well conserved in each group of Echinometridae and that the abundance of spines and the size and shape of the body skeleton changed relatively frequently and hence were less conserved. In Echinometrid species, morphological variation in relatively unconserved features tends to be associated with latitudinal distributions, rather than phylogenetic relationships, indicating that the morphological diversity of larval skeletons could have been caused by adaptation to the habitat environment. Some morphological differences, however, seem to be nonfunctional and generated by the constraints on larval skeletogenesis. Thus, echinometrid species can be a good model with which to study the evolutionary history from both ecological and developmental standpoints.     

Juvenile production of the red sea urchin Strongylocentrotus franciscanus (Echinodermata: Echinoidea) in Baja California, Mexico

The red sea urchin Strongylocentrotusfranciscanus (Agassiz 1863) is harvested commercially in Baja California, Mexico, since 1970; however, in the last ten years the capture per unit effort (CPUE) has decreased from 310 kg/fishing unit/day to 120 kg/fishing unit/day. For this reason, actions were taken to develop a culture technology allowing massive production of juveniles for re-stocking natural populations or for growing them commercially. We summarize some of the basic studies and main achievements in this effort. In Baja California, considerably faster larval development (approximately 21 days) has been attained than in the US northwest coast (62 days). Spawning of red sea urchins was routinely induced with KCI while egg fertilization was performed using a 100,000-sperm/ml solution. Six microalgae species were tested and Rhodomonas sp. produced the best larval development. The mean survival rate at the end of the larval period was 25%, but results varied widely with bactch. From the feed ratios tested, best results were obtained using 7000 cel/ml during the first week of larval development, followed by 10,000 cel/ml during the second and 15,000 cel/ml during the third week. KCl proved the most consistent metamorphic inducer, regularly yielding metamorphosis percentages higher than 90%. Metamorphosis was considered complete when the functional jaw that juveniles use for first benthic feeding appeared (as soon as 20 days after induction). With this method several thousands of red sea urchin juveniles were produced. They reached up to 1.5 mm in size during the first 50 days of culture after metamorphosis, showing the great potential for mass production of this species in the laboratory.     

Larvae and postlarvae production of the green-white sea urchin Lytechinus variegatus (Echinodermata: Echinoidea) in culture conditions

We evaluated the biological feasibility of massive larvae and postlarvae production of the Caribbean green-white urchin Lytechinus variegatus. Experiments were designed to choose the initial larval density and microalgae diets under culture, to study metamorphosis, postlarval and juvenile growth. Massive production of competent larvae 650 microm long at 12-13 days is possible using larval densities of 0.25 to 1 larva/ml. The microalgae Rhodomonas sp. was suitable for the optimization of larval growth and survival. Metamorphosis of 100% of the larvae can be induced with films of bentic diatoms (Navicula sp. and Amphora sp.), after 96 h; however, diatoms are not adequate for postlarval development and a food supply of Ulva lactuta is necessary for proper growth. For juveniles, a diet of macroalgae (U. lactuta) and/or commercial marine shrimp culture pellet food is enough for growth, but the best results were obtained with shrimp or U. lactuta used alone (85-86%, against 46% with the mixed diet). We recommend future experiments on nutritional requirements to optimize growth of these and subsequent stages.     

Identification of protein components from the mature ovary of the sea urchin Evechinus chloroticus (Echinodermata: Echinoidea)

The gonads of sea urchins are a high value seafood product, with considerable research being undertaken worldwide on the development of sea urchin aquaculture. As the best prices are obtained for specific gonad attributes, research has also focused on the development of artificial diets that enhance gonad quality and quantity. Total protein has been used as a measure of gonad quality; yet no studies to date have applied proteomics technology to diet development. Here we use a MudPIT and 2-DE approach to describe the major proteins in mature ovaries of a New Zealand sea urchin Evechinus chloroticus. This tissue, which is a target seafood product, contained 138 proteins that were identified from the recently completed sea urchin genome (Strongylocentrotus purpuratus) with high confidence. The majority of these proteins had general functions, with only 12 related to ovarian reproductive function. Eighteen proteins were located on the 2-DE; four of these were directly identified from S. purpuratus protein sequences. In combination this paper shows that the genome resources of S. purpuratus can be used to identify proteins in sea urchins from different families; describes the proteome of E. chloroticus mature ovary; and, provides proteomic tools for analysis of gonads from other edible sea urchins.     

Gametogenesis correlated with steroid levels during the gonadal cycle of the sea urchin Paracentrotus lividus (Echinodermata: Echinoidea)

The specific mechanism regulating reproduction in invertebrates is a field of topical interest which needs to be explored in detail considering also the intriguing possible comparison with vertebrates. In this paper levels of Testosterone (T) and Estradiol (E2) and their reciprocal ratios were determined in ovaries and testis of the echinoid model species Paracentrotus lividus during the year 2004 by taking into account a putative relationship between steroid levels and reproductive cycle. T levels appeared to significantly vary during male reproductive cycle, thus suggesting a possible role of this hormone in regulation of spermatogenesis as demonstrated for other echinoderms. E2 levels were lower in males with respect to females; consequently E2 involvement in oogenesis is hypothesized. In parallel with steroid levels evaluation, variations in P450-aromatase activity and its possible role on regulation of gametogenesis were also considered. Clear correlations between steroid levels and gonad index (GI), as well as between GI and reproductive cycle were not detected, suggesting that GI alone is not a reliable parameter in describing the reproductive status of the gonads. Altogether the results obtained so far confirm the presence of a relationship between steroid levels and reproductive cycle as suggested by previous results on different echinoderm species.     

Genetic diversity and population structure of the commercially harvested sea urchin Paracentrotus lividus (Echinodermata, Echinoidea)

The population structure of the edible Atlanto-Mediterranean sea urchin Paracentrotus lividus is described by analysing sequence variation in a fragment of the mitochondrial gene cytochrome c oxidase subunit I in 127 individuals from 12 localities across south-west Europe. The study revealed high levels of genetic diversity but low levels of genetic structure, suggesting a large degree of gene flow between populations and panmixis within each, the Mediterranean and Atlantic basins. However, we found significant genetic differentiation between the two basins probably due to restricted gene flow across the geographical boundary imposed by the area of the Strait of Gibraltar. Populations of P. lividus appeared to have experienced a recent demographic expansion in the late Pleistocene. We provide new evidence on the population structure of this commercial species, predicting a healthy stock of this sea urchin on the Mediterranean and Atlantic coasts.     

Pourtalesiid sea urchins (Echinodermata: Echinoidea) of the northern Mid-Atlantic Ridge

Five species of deep-sea pourtalesiid echinoids were collected by the RV G.O. Sars MAR-ECO expedition to the northern part of the Mid-Atlantic Ridge: Solenocystis imitans new genus, new species, Echinosigra phiale (Thomson, 1872), Echinosigra (Echinogutta) fabrefacta Mironov, 1974, and two unidentified species of Pourtalesia. Three types of plastron plating are distinguished within the family Pourtalesiidae; in Solenocystis new genus the plating is similar to that in Spatagocystis, a genus known only from the deep-sea Antarctic. Each of three groups with distinctive plastron plating shows a continuous morphological gradation from a less elongated test without subanal rostrum to an extraordinarily elongated test with large rostrum and wide subanal fasciole; the ratio width/length of test varies from 0.86 to 0.12. Some morphological characters of the pourtalesiids are regarded as adaptations to burrowing and appear to be correlated with test elongation. The family is represented in the northern Atlantic only by 'advanced' genera (Echinosigra, Pourtalesia and Solenocystis) with very elongated tests.     

Ultrastructural evidence for Sertoli-like cells in the testis of the asteroid,Luidia clathrata (Say) (Echinodermata: Platyasterida)

Non-germinal cells arise adjacent to the basal lamina and extend between the numerous germinal celli. Nuclei of these non-germinal cells may be positioned near the basal lamina or more peripherally between the spermatocytes. Thin cytoplasmic processes extend between the spermatocytes to the spermatids. These cytoplasmic processes vary in electron density from the cytoplasm of the germinal cells. These non-germinal cells closely resemble the vertebrate Sertoli cell.     

Comparative morphology of the axial complex and interdependence of internal organ systems in sea urchins (Echinodermata: Echinoidea)

Background

The axial complex of echinoderms (Echinodermata) is composed of various primary and secondary body cavities that interact with each other. In sea urchins (Echinoidea), structural differences of the axial complex in "regular" and irregular species have been observed, but the reasons underlying these differences are not fully understood. In addition, a better knowledge of axial complex diversity could not only be useful for phylogenetic inferences, but improve also an understanding of the function of this enigmatic structure.

Results

We therefore analyzed numerous species of almost all sea urchin orders by magnetic resonance imaging, dissection, histology, and transmission electron microscopy and compared the results with findings from published studies spanning almost two centuries. These combined analyses demonstrate that the axial complex is present in all sea urchin orders and has remained structurally conserved for a long time, at least in the "regular" species. Within the Irregularia, a considerable morphological variation of the axial complex can be observed with gradual changes in topography, size, and internal architecture. These modifications are related to the growing size of the gastric caecum as well as to the rearrangement of the morphology of the digestive tract as a whole.

Conclusion

The structurally most divergent axial complex can be observed in the highly derived Atelostomata in which the reorganization of the digestive tract is most pronounced. Our findings demonstrate a structural interdependence of various internal organs, including digestive tract, mesenteries, and the axial complex.     

Morphological and ultrastructural characterization of the alimentary canal in larvae of Streltzoviella insularis (Staudinger) (Lepidoptera: Cossidae)

Streltzoviella insularis (Staudinger) is an important tree‐boring pest, that primarily damages Sophora japonica (Linnaeus) and Ginkgo biloba (Linnaeus), as well as other common species, at great economic cost to the urban landscape construction industry in China. In the present study, the alimentary canal morphology of S. insularis was observed using light microscopy, and its ultrastructure was investigated by scanning and transmission electron microscopy. The foregut of S. insularis can be divided into the pharynx, esophagus, crop, proventriculus, and cardiac valve. The well‐developed crop forms the longest section of the foregut. It is able to store large amounts of food and is lined with a monolayer of epithelial cells. Many sclerotized microspines occur on the surface of the anterior intima and there are dense spines on the posterior intima of the proventriculus. Epithelial cells of the midgut include columnar cells, goblet cells, and regenerative cells, but endocrine cells are absent. The hindgut consists of the pyloric valve, ileum, and rectum. There is no clear distinction between the ileum and colon. The intima surface of the pyloric valve carries many microspines, whereas the intestinal wall of the rectum is thin with well‐developed rectal pads. The rectal epithelial cells form a squamous monolayer. A cryptonephric excretory system is located in the hindgut. There are six spiral Malpighian tubules, in which a cellular layer on a basement membrane encloses a lumen. These results will provide the basis for further studies of the structure and function in S. insularis larvae.     

Phylogeography of the sea urchin Paracentrotus lividus (Lamarck) (Echinodermata:Echinoidea): first insights from the South Tyrrhenian Sea

The sea urchin Paracentrotus lividus (Lamarck) (Echinodermata: Echinoidea) is an Atlanto-Mediterranean species abundant in the littoral zone, where it occurs in the sublittoral down to 20 m. The aim of our work is to investigate the genetic patterns of P. lividus along the South Tyrrhenian coasts. Five specimens were collected in six localities, from the Gulf of Naples and the Cilento coast. The nuclear rDNA ITS2 spacer and the two mitochondrial genes 16S and COI were used for the analysis. The three markers utilised did not show any structure among populations from the Gulf of Naples. All populations appear to be polyphyletic, with Cilento samples more differentiated from the others. This suggests the existence of phylogeographic structure at larger geographic scale. Absence of genetic structure has been interpreted taking into consideration theoretical dispersal of the planktotrophic larvae, which can survive for 4–8 weeks before settlement, marine current patterns and persistence of the species in the area.     

Gonadal growth and reproduction in the commercial sea urchin Paracentrotus lividus (Lamarck, 1816) (Echinodermata: Echinoidea) from southern Spain

The reproductive state of six populations of the commercial sea urchin P. lividus was investigated. Specimens were sampled monthly from six sites of southern Spain. The development of the gonad during the period of study was measured using histological methods and categorised by four stages for females and two for male specimens. A Gonadosomatic Index was also established. The six populations exhibited similar reproductive patterns, having mature gonads during almost all months while reconstitution and spent were present mainly in autumn. The population from Sancti Petri had the highest Gonadosomatic Index value. Higher food availability may explain this difference.     

Evolution of a novel muscle design in sea urchins (Echinodermata: Echinoidea)

The sea urchin (Echinodermata: Echinoidea) masticatory apparatus, or Aristotle's lantern, is a complex structure composed of numerous hard and soft components. The lantern is powered by various paired and unpaired muscle groups. We describe how one set of these muscles, the lantern protractor muscles, has evolved a specialized morphology. This morphology is characterized by the formation of adaxially-facing lobes perpendicular to the main orientation of the muscle, giving the protractor a frilled aspect in horizontal section. Histological and ultrastructural analyses show that the microstructure of frilled muscles is largely identical to that of conventional, flat muscles. Measurements of muscle dimensions in equally-sized specimens demonstrate that the frilled muscle design, in comparison to that of the flat muscle type, considerably increases muscle volume as well as the muscle's surface directed towards the interradial cavity, a compartment of the peripharyngeal coelom. Scanning electron microscopical observations reveal that the insertions of frilled and flat protractor muscles result in characteristic muscle scars on the stereom, reflecting the shapes of individual muscles. Our comparative study of 49 derived "regular" echinoid species using magnetic resonance imaging (MRI) shows that frilled protractor muscles are found only in taxa belonging to the families Toxopneustidae, Echinometridae, and Strongylocentrotidae. The onset of lobe formation during ontogenesis varies between species of these three families. Because frilled protractor muscles are best observed in situ, the application of a non-invasive imaging technique was crucial for the unequivocal identification of this morphological character on a large scale. Although it is currently possible only to speculate on the functional advantages which the frilled muscle morphology might confer, our study forms the anatomical and evolutionary framework for future analyses of this unusual muscle design among sea urchins.     

Biodiversity and historical biogeography of stalked crinoids (Echinodermata) in the deep sea

About 95 species of stalked crinoids are now described from 60m to hadal depths, but our knowledge remains far from complete. Depending on which species concept is used, estimates of species richness can be dramatically different. It is necessary to have a homogeneous concept for taxonomic units. The abundance of the crinoid fossil record allows a discussion of the ancestry of deep sea crinoid fauna. Stalked crinoids have a horizontal diversity pattern with three regional centres of high diversity (i.e. western tropical Pacific, western tropical Atlantic and north-eastern Atlantic). Vertical patterns show two faunal strata which vary in importance among provinces. The epibathyal stratum has apparently remained relatively similar in intertropical areas since the Mesozoic. Despite environmental changes related to glaciation since the Middle Miocene, the deepest crinoid fauna (i.e. the deep sea fauna sensu stricto at depths more than 1000 ± 200 m) have a very ancient origin with a dispersion closely related to plate tectonics. The bathyal fauna on hard substrates includes a few living fossils and has a high historical interest.     

Mechanical properties and structure of the spine-joint central ligament of the sea urchin, Diadema setosum (Echinodermata, Echinoidea)

In the regular echinoid Diadema setosum Leske the central ligament that connects the spine to its tubercle is mainly composed of closely-packed collagen fibres which are arranged in the longitudinal axis of the ligament. The mechanical properties of the ligament are quite different from ligament to ligament: the viscosity determined by creep tests ranges from 0·02 to 6 GPa·s. The viscosity of a ligament changes reversibly in response to stimulation. Adrenaline and noradrenaline (10^-6-10^-3 M) decrease the viscosity. Acetylcholine (10^-8-10^-3 M), artificial sea water containing high (100 MM) potassium concentration, and electrical stimulation increase the viscosity. As the ligament contains no muscle cells in it, the viscosity change cannot be attributable to muscle activities. The functional significance of having a central ligament with variable and controllable viscosity is that it binds the spine base onto the articulation surface of the tubercle, so as to reduce the possibility of spine dislocation, while being flexible enough to allow the spine considerable freedom of movement when necessary.     

Effects of inorganic and organic phosphate exposure on aspects of reproduction in the common sea urchin Lytechinus variegatus (Echinodermata: Echinoidea)

The common nearshore sea urchin Lytechinus variegatus is capable of surviving exposure to inorganic phosphate concentrations as high as 3.2 mg/l(-1) and organic phosphate concentrations of 1,000 mg/l(-1). However, chronic exposure to low, medium, or high sublethal concentrations of these phosphates inhibits gonadal tissue indices and spawning activity while altering biochemical composition of gonads, reducing size frequencies of oocyte diameters, and changing gonadal volume fractions. Gonad indices declined significantly in individuals maintained in all phosphate concentrations after both one- and two-month exposures, while percentages of ripe individuals (oozing gametes upon dissection) were reduced after a two-month exposure in individuals maintained in medium and high organic phosphate concentrations. Levels of carbohydrates and lipids were lower in gonads of individuals maintained in all concentrations of both phosphates. Size frequency distributions of oocyte diameters revealed a dramatic decrease in oocyte size with increasing concentrations of both phosphates. Gonadal volume fractions of developing male and female gametes decreased with exposure to increasing phosphate levels. Volume fractions of nutritive phagocytes declined in testes of individuals held in the highest concentration of organic phosphate but displayed no significant change in ovaries. Volume fractions of mature gametes also decreased in gonads of individuals exposed to increasing concentrations of inorganic phosphate, but they remained constant in individuals exposed to all concentrations of organic phosphate. These findings indicate that shallow-water populations of L. variegatus subjected to inorganic and organic phosphate pollutants will exhibit stress that may impair reproductive output, gametogenesis, and spawning in the natural environment.     

Phylogeography and genetic structure of the edible sea urchin Paracentrotus lividus (Echinodermata: Echinoidea) inferred from the mitochondrial cytochrome b gene

Phylogeographical analysis of Paracentrotus lividus was carried out by means of sequencing the mitochondrial cytochrome b gene (1143 bp) of 260 individuals collected at 22 Mediterranean and four Atlantic localities. Against a background of high haplotype diversity and shallow genetic structuring, we observed significant genetic divergence between the Adriatic Sea and the rest of the Mediterranean, as well as between the Mediterranean and the Atlantic sample groups. Furthermore, on the largest spatial scale, isolation by distance was detected. Three main haplogroups were identified by network and Bayesian assignment analyses. The relative proportions of haplogroups were different in the four regions considered, with the exception of Western and Eastern Mediterranean that showed a similar pattern. This result together with the outcome of S_nn statistics, analysis of molecular variance and network analyses allowed to identify three weakly differentiated populations corresponding to the Atlantic, Western + Eastern Mediterranean, and Adriatic seas. Analyses of mismatch distribution and neutrality tests were consistent with the presence of genetic structuring and past demographic expansion(s). From a fisheries perspective, the results obtained in the present study are consistent with genetic sustainability of current exploitation; local depleted stocks are recurrently replenished by recruits that may have originated from nonharvested areas. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 910–923.