Larval growth and survival of Echinometra lucunter (Echinoidea: Echinometridae) fed with microalgae Chaetoceros gracilis and Isochrysis galbana

Thirty sexually mature sea urchins (Echinometra lucunter; diameter 45.8 +/- 17.5 mm) were collected at Macanao, Margarita Island, Venezuela (11 degrees 48'29" N / 64 degrees 13'10" W). They were injected potassium chloride (50 M) directly into the celomic cavity. After two minutes 90% spawned (17 females and 10 males), the others never spawned. Fertilization was 87.0 +/- 12.6% (1:100 oocytes/sperm) at 29 +/- 2 degrees C. The fertile eggs were placed in three treatment gropsu with nine containers (18 liters; 2 eggs/ml) each, all with bottom aeration. Treatments were: Chaetoceros gracilis; Isochrysis galbana, and a mixture of both microalgae (respectively: 20 000 and 60,000 cell/ml for each microalgae, 1:1 for the mixture). Salinity, pH, temperature and larval survival were determinated daily. The study ended when the post-metamorphic phase was completed. The embryonic development time was 16.3 +/- 0.2 h until the prism stage at pH 8.4 +/- 0.1; 38 +/- 1 psu and 28 +/- 1.4 degrees C. The two-arms larval stage was reached at 24 h: 33 min, with a total length of 190 +/- 16.3 microm fed on C. gracilis, 152 +/- 19.0 microm with I. galbana and 182.4 +/- 14.1 microm with the mixture. The larvae next to metamorphosis reabsorbed the arms and had the characteristic shape of juvenile urchins at 12 days with 670.2 +/- 22.2 microm fed on C. gracilis, 665 +/- 12.1 microm fed on I. galbana and 670 +/- 14.1 microm fed on the mixture. The accumulated survival to the juvenile stage was 14.7 +/- 3.8% when fed on C. gracilis, higher than the other treatments (5.4 +/- 1.2; 14.0 +/- 2.6). E. lucunter is an excellent prospect to be commercially cultured because of its short embryonic (16 hours) and larval development time (12 days) and good survival rate when fed on monoculture (C. gracilis and I. galbana) or mixed diet (we recommend C. gracilis).     

Phagostimulation of Lytechinus variegatus (Lamarck) (Echinodermata: Echinoidea)

Artificial agarose foods containing either starch, glycogen, carragheenan, alginate, casein, gelatin, albumin, or peptones at a concentration of 1% dry weight were not phagostimulatory. Artificial foods containing sugars, especially galactose, at a concentration of 0.005 M were phagostimulatory. Of the common amino acids, only L‐phenylalanine was phagostimulatory at this concentration. The initiation of feeding postures (movement of the spines, tube feet, peristome, and Aristotle's lantern) of Lytechinus variegatus was little affected by the concentration of plant extracts. Many aspects of this behavior were attributable to physical rather than chemical stimulation. Feeding rates of L. variegatus upon artificial agarose foods containing either Thalassia testudinum or Gracilaria verrucosa were directly dependent upon plant tissue concentration. All artificial agarose food‐items were initially moved to the mouth and fed upon, however the time spent feeding was directly dependent upon plant type and tissue concentration. Retention of the artificial food‐items upon the test after feeding was not influenced by plant type or tissue concentration. Differences in consumption of foods by L. variegatus are determined by the degree of persistence in feeding upon encountered items and not be selection before initiating feeding or by retention of food‐items for subsequent feedings.     

Insemination of immature sea urchin (Arbacia punctulata) eggs

Nuclei from osmotically opened erythrocytes and erythroblasts were injected into nucleated or enucleated Xenopus laevis eggs. Although the cleavage pattern of the recipient eggs which started to divide was normal in about half of the cases, nuclei from erythrocytes injected into nucleated or enucleated eggs never promoted development beyond the early gastrula stage. In contrast, nuclei from osmotically opened erythroblasts injected into enucleated eggs promoted development to early tadpole stages (stages 29–36). Frequently, injection of osmotically broken erythroblasts injected into nonenucleated eggs gave rise to triploid larvae which all died at roughly the same early tadpole stages (29–36). Surprisingly, development did not proceed to the stage of advanced organogenesis (stages 44–47), which is easily reached by gynogenetic haploids: The presence of the haploid genome derived from the egg pronucleus did not significantly improve the developmental capacity. Embryos obtained by single injection of erythrocyte nuclei into nucleated eggs were unable to pass the gastrula stage. To invalidate the interpretation that the observed arrest in development was related to nuclear damage during injection of the recipient eggs, single unbroken erythrocytes and unbroken erythroblasts were transferred into nucleated and enucleated eggs. No cleavage was observed in both classes of eggs injected with unbroken erythrocytes. In contrast, erythroblasts were found to induce cleavage in the recipient eggs at a frequency of about 11%. To ascertain that the nucleus of unbroken erythroblasts participated in development, the 1-nucleolar marker was used. Diploid embryos with only one nucleolus present were found following injection of unbroken erythroblasts into enucleated eggs from 2nu females. Triploid 2nu embryos were detected following injection of (diploid) 1nu erythroblasts into nonenucleated eggs from 2nu females. The most advanced development stages reached by these embryos did not, however, differ from the best results found in the first class of experiments: Nuclei from erythroblasts injected undamaged into nucleated or enucleated eggs never developed into a normal tadpole. Serial transfer experiments were performed using normally gastrulating embryos which had developed, following the injection of 1nu unbroken erythroblasts into recipient eggs. These donors for serial transfer experiments were checked for the presence of the 1nu marker. In addition they had passed through a normally cleaving eight-cell stage. No improvement in developmental capacity as compared to first transfer experiments could be found.     

Echinoids (Echinodermata: Echinoidea) from the Gulf of Mexico

The echinoid fauna of the Gulf of Mexico collected during three research cruises (20-1260 m depth) was surveyed from samples were taken at 43 stations. A total of 190 individuals were identified (eight orders, 11 families, 15 genera and 18 species). Six species are new records for the Gulf of Mexico: Stylocidaris lineata, Phormosoma placenta placenta, Plesiodiadema antillarum, Plethotaenia spatangoides, Brissopsis atlantica and Hypselaster limicolus. This adds to the little information available on the echinoid fauna of Tamaulipas, Veracruz, Tabasco, Campeche and Yucatan states in Mexico.     

Behavioral ecology of Strongylocentrotus droebachiensis (Muller) (Echinodermata: Echinoidea)

Summary Field observation indicated that extensive aggregation behavior occurs in Strongylocentrotus droebachiensis, an important echinoid herbivore in rocky subtidal zones of northern New England. The relationship of this behavior to the behavioral ecology of this species was investigated. Laboratory studies indicated that urchins are chemically sensitive to the stimuli of food and other urchins. Quantitative field experiments with tagged animals and transects showed the existence of two types of aggregations: feeding and nonfeeding, which appear to be pervasive phenomena in the biology of S. droebachiensis.It is concluded that, depending upon their hunger state, urchins shuttle between feeding aggregations in exposed locations and non-feeding aggregations located in relatively sheltered areas (where they spend most of their time). Individuals of S. droebachiensis aggregate actively through response to other urchins in the form of chemotaxis. The aggregative behavior may be maintained for its selective value in terms of defense from predation and, to lesser extents, facilitation of feeding and breeding.     

Surface changes at fertilization: integration of sea urchin (Arbacia punctulata) sperm and oocyte plasma membranes

To examine the integration and fate of the sperm plasma membrane following its incorporation into the oocyte plasma membrane, we have fertilized sea urchin (Arbacia punctulata) gametes reciprocally labeled with cationized ferritin. When unlabeled oocytes were inseminated with labeled sperm, cationized ferritin acceptors moved laterally from the sperm plasma membrane into the fertilization cone and surrounding microvilli, mixing with components of the oocyte plasmalemma. Labeled oocytes inseminated with unlabeled sperm produced extremely large fertilization cones, completely devoid of cationized ferritin, while the remainder of the oocyte surface remained heavily labeled. Surface area measurements indicated that if all the sperm plasmalemma were utilized to delimit a fertilization cone it would provide less than 10% of the total surface membrane. Evidence is presented indicating that a principal source of membrane to the expanding fertilization cone of inseminated oocytes is from microvilli, i.e., microvilli are retracted to accommodate fertilization cone formation. Membrane delimiting the fertilization cone has a much lower affinity for agents (cationized ferritin and concanavalin A) that stain negatively charged and carbohydrate moieties compared to other regions of the oocyte surface. These ultrastructural observations indicate that significant rearrangements occur in the oocyte and sperm plasma membranes following gamete fusion which give rise to asymmetries in membrane topography; components of both membranes are redistributed within the bilayer adjacent to and delimiting the fertilization cone.     

Phylogeny of Early Cretaceous spatangoids (Echinodermata: Echinoidea) and taxonomic implications

A phylogenetic analysis of 36 species provides a test for the taxonomy and the history of Early Cretaceous spatangoids. Most taxonomic units from genera to suborders are consistent with the proposed phylogenetic framework. We retain Hemiasterina, Micrasterina, Hemiasteridae, Schizasteridae, Hemiaster , Heteraster , Mecaster , and Periaster as original monophyletic groups. However, all of these clades originate without the classical apomorphies normally ascribed to them. We suggest a revision of their diagnoses and of the generic attributions of basal species. Some ill-defined, 'primitive', and paraphyletic taxa are recognised: Toxaster , Epiaster , Palhemiaster , and Toxasteridae. Even if they do not have phylogenetic meaning, they are retained here, pending a more complete revision.     

Effect of ultraviolet radiation on growth and percent settlement of larval Lytechinus variegatus (Echinodermata: Echinoidea)

Lytechinus variegatus larvae were used to examine the effects of ecologically relevant short-term exposures of ultraviolet radiation (UVR) on larval morphology and settlement. A laboratory experiment tested the blastula, gastrula, and early pluteus stages of larval development for susceptibility to damage from exposure to UVR produced by an artificial light source. Larval post-oral arm length and percent settlement were measured to assess UVR effects. Larvae exposed at the blastula and gastrula stages had a reduction in larval post-oral arm length compared with non-exposure controls, while larvae exposed at the early pluteus stage were similar to controls. Larvae exposed in the gastrula stage had a significant reduction in percent settlement compared with controls. Negative consequences from ecologically relevant levels of UVR were differentially dependent on the larval stage at time of exposure suggesting time point in larval development at exposure will be important in interpreting effects on higher levels of biological organization.     

Organization of microfilaments in sea urchin (Arbacia punctulata) eggs at fertilization: Effects of cytochalasin B

Investigations were carried out to examine more closely the aggregations of microfilaments associated with the elongation of microvilli and formation of fertilization cones and the effects of cytochalasin B (CB) on these processes in Arbacia eggs following insemination. At 1 to 5 min postinsemination fertilized eggs were treated with 1–10 μg/ml CB and then prepared for electron microscopy at periodic intervals. Examination of CB-treated and untreated specimens demonstrated that: (1) Reorganization of the egg's microvilli took place soon after insemination; this process, as well as formation of fertilization cones, was correlated with the appearance of fascicles of microfilaments. (2) CB inhibited the formation of fertilization cones and the elongation of microvilli. Bundles of microfilaments were not observed in CB-treated zygotes. (3) CB prevented the normal movements (rotation) of the incorporating spermatozoon into the egg cortex but did not inhibit the migration or fusion of the male and female pronuclei.     

Phylogeny of Cidaroida (Echinodermata: Echinoidea) based on mitochondrial and nuclear markers

We present the first molecular phylogeny of Cidaroida, one of the most problematic groups within the echinoids. Two genes??the nuclear ribosomal gene 28?S rRNA and the mitochondrial protein-encoding gene COI??were obtained from 21 specimens representing 17 genera and 20 species, among which 13 species belong to Cidaroida. Phylogenetic analyses of the combined molecular data using parsimony and maximum likelihood optimality criteria resulted in a well-resolved phylogeny. Our results are broadly compatible (with the notable exception of Cidaris cidaris) with previous results obtained from morphological data. We find that Cidaroida represent a monophyletic group sister to the non-cidaroid Echinoidea. The family Cidaridae sensu Mortensen (1928) and Fell (1966) is paraphyletic because of the placement of Psychocidaris ohshimai as sister-group to Histocidaris elegans. Inside the Stylocidarina, we show that the two Atlantic species Stylocidaris affinis and Stylocidaris lineata constitute a well-supported clade. However, these two taxa could also represent two morphotypes within a single species showing high morphological variation.     

Histology of the globiferous pedicellariae of Psammechinus miliaris (Echinodermata: Echinoidea)

The general morphology, function and histology of the globiferous pedicallariae of the regular sea-urchin, Psammechinus miliaris are described. There is a ganglion under the sensory epidermis on the inner surface of each jaw. This is the first discovery of a ganglion in any echinoid.
The regeneration of globiferous pedicellariae in laboratory conditions takes 25 to 40 days.     

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.     

Thyroid hormones determine developmental mode in sand dollars (Echinodermata: Echinoidea)

Evolutionary transitions in larval nutritional mode have occurred on numerous occasions independently in many marine invertebrate phyla. Although the evolutionary transition from feeding to nonfeeding development has received considerable attention through both experimental and theoretical studies, mechanisms underlying the change in life history remain poorly understood. Facultative feeding larvae (larvae that can feed but will complete metamorphosis without food) presumably represent an intermediate developmental mode between obligate feeding and nonfeeding. Here we show that an obligatorily feeding larva can be transformed into a facultative feeding larva when exposed to the thyroid hormone thyroxine. We report that larvae of the subtropical sand dollar Leodia sexiesperforata (Echinodermata: Echinoidea) completed metamorphosis without exogenous food when treated with thyroxine, whereas the starved controls (no thyroxine added) did not. Leodia sexiesperforata juveniles from the thyroxine treatment were viable after metamorphosis but were significantly smaller and contained less energy than sibling juveniles reared with exogenous food. In a second starvation experiment, using an L. sexiesperforata female whose eggs were substantially larger than in the first experiment (202+/-5 vs. 187+/-5 microm), a small percentage of starved L. sexiesperforata larvae completed metamorphosis in the absence of food. Still, thyroxine-treated larvae in this experiment completed metamorphosis faster and in much higher numbers than in the starved controls. Furthermore, starved larvae of the sand dollar Mellita tenuis, which developed from much smaller eggs (100+/-2 microm), did not complete metamorphosis either with or without excess thyroxine. Based on these data, and from recent experiments with other echinoids, we hypothesize that thyroxine plays a major role in echinoderm metamorphosis and the evolution of life history transitions in this group. We discuss our results in the context of current life history models for marine invertebrates, emphasizing the role of egg size, juvenile size, and endogenous hormone production for the evolution of nonfeeding larval development.     

Abundance of Diadema antillarum (Echinodermata: Echinoidea) in the coasts of Venezuela

Diadema antillarum is a shallow-water sea-urchin from the tropical Atlantic whose populations almost disappeared in 1983-84 because of widespread mortalities which reached 87-100 %. In Venezuela, urchin population densities before the mortality event were comparable to those of other Caribbean regions; however, later abundancies remain unknown. The objectives of this study were to evaluate the recent densities of certain D. antillarum populations along the Venezuelan coasts and compare the densities at the Parque Nacional Mochima before and after the mortality. At each location urchin densities were determined by means of transects using 1 m2-quadrats as sampling units. The highest mean densities were observed at the sites on the central coast: Ensenada de Oricao, 0.28 ind/m2 (2002) and 1.05 ind/m2 (2003), and Chichiriviche de la Costa, 0.84 ind/m2 (2002) and 0.74 ind/m2 (2003). In Mochima, the mean density before the mortality for D. antillarum oscillated between 0.28 and 4 ind/m2, after the mortality event the mean density varied between 0.15 ind/m2 (2000) and 0.47 ind/m2 (2000). The populations of D. antillarum studied at Parque Nacional Morrocoy and Refugio de Fauna Silvestre Cuare showed highest densities at Playuela (0.43 ind/m2) and Cayo Sur (0.95 ind/m2) respectively, whereas other sites showed densities below 0.1 ind/m2. The density registered at Playuela in 2003 is lower than that reported before the mortality event (0.58-3.64 ind/m2). The density for Parque Nacional Archipiélago de Los Roques, specifically for the Arrecife de Herradura remained constant between 2002 and 2003 with values between 0.22-0.23 ind/m2 respectively. To conclude, the sea urchin abundancies observed at most of the Venezuelan coastal sites that we studied were higher than those reported for other areas of the northern Caribbean, even though the values have not yet returned to those preceding the 1984 mass-mortality event, due to the slow recovery of the populations.     

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.     

Larval development of the featherstar Aporometra wilsoni (Echinodermata: Crinoidea)

Abstract. Larval development of a small ovoviviparous comatulid crinoid, Aporometra wilsoni , was investigated using a population from South Australia. The genital pinnules of reproductive females each contain an ovary, within which are oocytes of various stages. Generally, one or more developing larvae lie outside the ovary, but within each pinnule. Larvae pass through "uniformly ciliated" and doliolaria stages before they exit the pinnule via the genital pore. The doliolariae lack the usual ciliary bands and are unable to swim. Doliolariae dissected from pinnules were followed through metamorphosis to the cystidean stage until the pentacrinoid larval stage. While previous reports on Aporometra have noted pentacrinoids attached to the female, virtually no pentacrinoids were found attached to any of the hundreds of adult females observed during this study. Females sampled from the mid-reproductive season (September and October) were found to bear >2500 developing eggs and larvae at a time. It appears that emerging doliolariae fall from the female and attach to other substrates to complete development. Aporometra is classified in Notocrinida with the Antarctic crinoid Notocrinus. This classification is based in part on their supposedly homologous larval brooding. However, the reproductive mechanism in A. wilsoni is quite different from Notocrinus , calling into question their current status as sister taxa.     

Reproductive patterns within sub-populations of Lytechinus variegatus (Lamarck) (Echinodermata: Echinoidea)

The annual reproductive cycle of the sea urchin Lytechinus variegatus (Lamarck) was examined in individuals collected from four habitats in the Anclote estuary near Tarpon Springs, Florida. Two periods of advanced gametogenic activity were apparent, but this bimodal pattern was not necessarily reflected by gonad growth curves. Differences in reproductive events among sub-populations were related primarily to patterns of gonad growth. Reproductive synchrony present during spring became less apparent as spawning and recovery within the sub-populations took place at differential rates during late-spring and summer. The final spawnout in late summer returned the sub-populations into gametogenic synchrony.     

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.