New techniques for creating parthenogenetic larvae of the sea urchin Lytechinus pictus for gene expression studies.

BACKGROUND: Sea urchins are model organisms for studying the spatial-temporal control of gene activity during development. The Southern California species, Lytechinus pictus, has a sequenced genome and can be raised in the laboratory from egg to egg in 4 to 5 months. RESULTS: Here, we present new techniques for generating parthenogenetic larvae of this species and include a gallery of photomicrographs of morphologically abnormal larvae that could be used for transcriptomic analysis. CONCLUSIONS: Comparison of gene expression in parthenogenotes to larvae produced by fertilization could provide novel insights into gene expression controls contributed by sperm in this important model organism. Knowledge gained from transcriptomics of sea urchin parthenogenotes could contribute to parthenogenetic studies of mammalian embryos.

Genomic insights of body plan transitions from bilateral to pentameral symmetry in Echinoderms.

Echinoderms are an exceptional group of bilaterians that develop pentameral adult symmetry from a bilaterally symmetric larva. However, the genetic basis in evolution and development of this unique transformation remains to be clarified. Here we report newly sequenced genomes, developmental transcriptomes, and proteomes of diverse echinoderms including the green sea urchin (L. variegatus), a sea cucumber (A. japonicus), and with particular emphasis on a sister group of the earliest-diverged echinoderms, the feather star (A. japonica). We learned that the last common ancestor of echinoderms retained a well-organized Hox cluster reminiscent of the hemichordate, and had gene sets involved in endoskeleton development. Further, unlike in other animal groups, the most conserved developmental stages were not at the body plan establishing phase, and genes normally involved in bilaterality appear to function in pentameric axis development. These results enhance our understanding of the divergence of protostomes and deuterostomes almost 500 Mya.

Inhibitory effects of neurotoxin ß-N-methylamino-L-alanine on fertilization and early development of the sea urchin Lytechinus pictus.

Neurotoxin ß-N-methylamino-L-alanine (BMAA) has been widely detected in diverse aquatic organisms and hypothesized as an environmental risk to neurodegenerative diseases in humans. However, the knowledge of its toxicity to marine organisms requires attention. In the present study, embryos and sperm of the sea urchin, Lytechinus pictus, were used to assess the toxicity of BMAA. Effects of BMAA on fertilization and development of sea urchin embryos were measured, and its impacts on efflux transport of sea urchin blastula were also assayed. Results demonstrated that the fertilization and development of embryos were significantly inhibited by high concentrations of BMAA above 300 µg L-1. The EC50 values indicated by active swimming larvae and total larvae numbers at 96 HPF (hours post fertilization) were 165 µg L-1 (1.4 µmol L-1) and 329 µg L-1 (2.8 µmol L-1), respectively. Additionally, sperm exposed to BMAA for 10 min significantly reduced the fertilization ratio of sea urchin eggs. However, the ABC transport activity on the cytomembrane of sea urchin blastula was not inhibited by the presence of BMAA at 50 µg L-1, even up to 500 µg L-1. Abnormal division and developmental malformations occurred at different developmental stages for sea urchin embryos exposed to BMAA at 500 µg L-1. The inhibitory effects of BMAA on sea urchin embryos were reported at the first time in this study, for which the toxicological mechanisms will be explored in future studies.

Effects of ocean warming and acidification on fertilization success and early larval development in the green sea urchin Lytechinus variegatus.

Ocean acidification and warming are predicted to affect the early life of many marine organisms, but their effects can be synergistic or antagonistic. This study assessed the combined effects of near-future (2100) ocean acidification (pH 7.8) and warming (+3 °C) on the fertilization, larval development and growth of the green sea urchin, Lytechinus variegatus, common in tropical reefs of Florida and the Caribbean. Acidification had no effect on fertilization, but delayed larval development, stunted growth, and increased asymmetry. Warming decreased fertilization success when the sperm:egg ratio was higher (1847:1), accelerated larval development, but had no effect on growth. When exposed to both acidification and warming, fertilization rates decreased, larval development accelerated (due to increased respiration/metabolism), but larvae were smaller and more asymmetric, meaning acidification and warming had additive effects. Thus, climate change is expected to decrease the abundance of this important herbivore, exacerbating macroalgal growth and dominance on coral reefs.

Evolución temporal del índice gonadal de los erizos marinos Paracentrotus lividus (Echinoida: Echinidae) y Diadema africanum (Diadematoida: Diadematidae) en las islas Canarias (España) / Temporal evolution of the gonad index of the sea urchins Paracentrotus lividus (Echinoida: Echinidae) and Diadema africanum (Diadematoida: Diadematidae) in the Canary Islands (Spain)

Resumen Con el fin de establecer las épocas óptimas de explotación como posible recurso marisquero de dos especies de erizos de mar en Canarias, se estudió la evolución temporal del índice gonadal en diferentes años y localidades de Paracentrotus lividus (abril de 2006 a marzo de 2008 en la isla de Tenerife y en dos localidades: Las Galletas y La Jaca; abril de 2006 a enero de 2009 en la isla de Gran Canaria y en dos localidades: Ojos de Garza y Gando), y de Diadema africanum (de enero de 2010 a mayo de 2011 en la isla de Gran Canaria y en tres localidades: Risco Verde, Arguineguín y Agaete). En el caso de P. lividus, que se encuentra en el límite sur de su distribución, se constató la presencia de varios picos máximos anuales en el índice gonadal, probablemente relacionados con la temperatura. Estas épocas de mayor índice gonadal se correspondieron con los estados de maduración de las gónadas. La principal época de madurez coincidió con el otoño y comienzo del invierno (agosto, octubre y diciembre). Existió emisión de gametos esporádicos en el año según la localidad, en abril, en verano (junio, julio o agosto), otoño (septiembre u octubre) e invierno (diciembre, enero o febrero). Posiblemente esta situación tan dispar sea debido a la exposición al hidrodinamismo de la zona y a la presencia o no de abundante alimento. Para D. africanum se presentó el máximo índice en los meses de mayo-junio, con prácticamente pocas variaciones anuales en la localidad situada más al sur (Arguineguín). En conclusión, las dos especies de erizos serían complementarias como recursos marisqueros, ya que no se superponen sus periodos de máximos IG (época de captura) en el año.

Abstract There are three main species of regular sea urchins in the Canary Islands. To establish the optimal fishing seasons for two of them, we studied the evolution of the gonadal index in several years and locations, of Paracentrotus lividus (April 2006 to March 2008 on Tenerife island in two locations: Las Galletas and La Jaca and from April 2006 to January 2009 in Gran Canaria island in two locations: Ojos de Garza and Gando), and Diadema africanum (January 2010 to May 2011 on Gran Canaria island in three locations: Risco Verde, Arguineguin and Agaete). In the case of P. lividus, located on the southern edge of their distribution, the presence of several annual maximum gonadal indez peak was observed, probably related to the temperature. These times of gonadal index increases corresponded to the stages of maturation. The main period of maturity coincided with the fall and early winter (August, October and December). There was sporadic emissions of gametes depending on location, in April, during the summer (June, July or August), autumn (September or October) and winter (December, January or February). This fragmented situation is possibly due to exposure to the hydrodynamic area's and food abundance. The maximum presence of D. africanum occurred in the months of May to June, with virtually no variations throughout the year, in the southernmost locality (Arguineguin). In conclusion, the two species of sea urchins would be complementary shellfish resources, as their periods of maximum GI (Gonadal Index, capture time) do not overlap during the year. Rev. Biol. Trop. 63 (Suppl. 2): 251-260. Epub 2015 June 01.

The effects of dietary silver on larval growth in the echinoderm Lytechinus variegatus.

Previous studies have demonstrated that the euryhaline copepod Acartia tonsa is extremely sensitive to dietborne silver (Ag) exposure, with a 20 % inhibition (EC(20)) of survival occurring when copepods are fed algae with 1.6 µg g(-1) dry weight (dw) Ag, corresponding to a waterborne Ag concentration of 0.46 µg l(-1) Ag. In contrast, 43 µg l(-1) Ag is required to elicit similar effects in copepods exposed to Ag by way of water. In the current study, we investigated whether another planktonic marine organism might also be sensitive to dietary Ag. Specifically, we tested larvae of the echinoderm, Lytechinus variegatus in an 18-day study in which larvae were continuously exposed to Ag-laden algae (Isochrysis galbana). After 7 days of exposure, no significant effects were observed on larval growth up to the highest concentration tested (10.68 µg g(-1) dw Ag in algae after exposure to 3.88 µg l(-1) waterborne Ag). After 18 days, significant effects were observed in all Ag treatments resulting in a lowest-observable effect concentration of 0.68 µg g(-1) dw Ag in algae and corresponding waterborne Ag concentration of 0.05-0.07 µg l(-1) Ag (depending on background Ag [see Results]). However, the dose-response relationship was quite flat with a similar level of growth inhibition (approximately 15 %) in all Ag treatments, resulting in an EC(20) of >10.68 µg g(-1) dw Ag in algae (>3.88 µg l(-1) Ag in water). This flat dose-response relationship is characteristic of dietary metal (silver, copper, cadmium, nickel, and zinc) toxicity to copepods as well, although the effect is slightly more robust (approximately 20-30 % inhibition of survival or reproduction). We conclude that echinoderm larvae may be similar to copepods in their sensitivity to dietary Ag, although a better understanding of the mechanisms underlying the apparent flat dose-response relationships is clearly needed.

Organic matrix-related mineralization of sea urchin spicules, spines, test and teeth.

The camarodont echinoderms have five distinct mineralized skeletal elements: embryonic spicules, mature test, spines, lantern stereom and teeth. The spicules are transient structural elements whereas the spines, and test plates are permanent. The teeth grow continuously. The mineral is a high magnesium calcite, but the magnesium content is different in each type of skeletal element, varying from 5 to 40 mole% Mg. The organic matrix creates the spaces and environments for crystal initiation and growth. The detailed mechanisms of crystal regulation are not known, but acidic and phosphorylated matrix proteins may be of special importance. Biochemical studies, sequencing of the complete genome, and high-throughput proteomic analysis have not yet provided insight into the mechanisms of crystallization, calcite composition, and orientation applicable to all skeletal elements. The embryonic spicules are not representative of the mature skeletal elements. The next phase of research will have to focus on the specific localization of the proteins and individual biochemistries of each system with regard to mineral content and placement.

On the formation and functions of high and very high magnesium calcites in the continuously growing teeth of the echinoderm Lytechinus variegatus: development of crystallinity and protein involvement.

Sea urchin teeth grow continuously and develop a complex mineralized structure consisting of spatially separate but crystallographically aligned first stage calcitic elements of high Mg content (5-15 mol% mineral). These become cemented together by epitaxially oriented second stage very high Mg calcite (30-40 mol% mineral). In the tooth plumula, ingressing preodontoblasts create layered cellular syncytia. Mineral deposits develop within membrane-bound compartments between cellular syncytial layers. We seek to understand how this complex tooth architecture is developed, how individual crystalline calcitic elements become crystallographically aligned, and how their Mg composition is regulated. Synchrotron microbeam X-ray scattering was performed on live, freshly dissected teeth. We observed that the initial diffracting crystals lie within independent syncytial spaces in the plumula. These diffraction patterns match those of mature tooth calcite. Thus, the spatially separate crystallites grow with the same crystallographic orientation seen in the mature tooth. Mineral-related proteins from regions with differing Mg contents were isolated, sequenced, and characterized. A tooth cDNA library was constructed, and selected matrix-related proteins were cloned. Antibodies were prepared and used for immunolocaliztion. Matrix-related proteins are acidic, phosphorylated, and associated with the syncytial membranes. Time-of-flight secondary ion mass spectroscopy of various crystal elements shows unique amino acid, Mg, and Ca ion distributions. High and very high Mg calcites differ in Asp content. Matrix-related proteins are phosphorylated. Very high Mg calcite is associated with Asp-rich protein, and it is restricted to the second stage mineral. Thus, the composition at each part of the tooth is related to architecture and function.

Small micromeres contribute to the germline in the sea urchin.

Many indirect developing animals create specialized multipotent cells in early development to construct the adult body and perhaps to hold the fate of the primordial germ cells. In sea urchin embryos, small micromeres formed at the fifth division appear to be such multipotent cells: they are relatively quiescent in embryos, but contribute significantly to the coelomic sacs of the larvae, from which the major tissues of the adult rudiment are derived. These cells appear to be regulated by a conserved gene set that includes the classic germline lineage genes vasa, nanos and piwi. In vivo lineage mapping of the cells awaits genetic manipulation of the lineage, but previous research has demonstrated that the germline is not specified at the fourth division because animals are fertile even when micromeres, the parent blastomeres of small micromeres, are deleted. Here, we have deleted small micromeres at the fifth division and have raised the resultant larvae to maturity. These embryos developed normally and did not overexpress Vasa, as did embryos from a micromere deletion, implying the compensatory gene regulatory network was not activated in small micromere-deleted embryos. Adults from control and micromere-deleted embryos developed gonads and visible gametes, whereas small micromere-deleted animals formed small gonads that lacked gametes. Quantitative PCR results indicate that small micromere-deleted animals produce background levels of germ cell products, but not specifically eggs or sperm. These results suggest that germline specification depends on the small micromeres, either directly as lineage products, or indirectly by signaling mechanisms emanating from the small micromeres or their descendants.

Flow-cytometric analyses of viability biomarkers in pesticide-exposed sperm of three aquatic invertebrates.

Toxicity studies on sperm often use fertilization success as the end point. This type of assay can be affected by sperm density, egg quality, and sperm-egg compatibility. Testing sperm viability biomarkers with flow cytometry is a fast, high-throughput technique for seminal analysis. In this study, we detected sperm viability biomarkers with several fluorescent reporter dyes using flow cytometry in three aquatic invertebrates (Crassostrea virginica, Dreissena polymorpha, and Lytechinus variegatus) after exposure to a pesticide and herbicide. The pesticide, Bayluscide, appeared to affect mitochondrial membrane potential in the sperm of all three species, as measured with MitoTracker Red CMXRos. A decrease in the percentage of sperm stained with SYBR-14 (indicating uncompromised plasma membrane) was observed in C. virginica and D. polymorpha sperm exposed to Bayluscide, but propidium iodide staining (indicating compromised plasma membranes) appeared to be inhibited by Bayluscide. Acrosome-reacted sperm, as measured by FITC-PNA, decreased after Bayluscide exposure in C. virginica and D. polymorpha sperm. The herbicide, Roundup Ready To-Use-Plus, did not affect the overall percentages of sperm stained with MitoTracker but did cause an increase in MitoTracker fluorescence intensity at 16 mg/L in D. polymorpha. Roundup also caused significant decreases in SYBR-14 staining, significant increases in propidium iodide staining, and significant increases in FITC-PNA staining in D. polymorpha sperm. By not having to rely on egg availability and optimal sperm density, sperm toxicity can be more accurately assessed with flow cytometry as being directly correlated to sperm viability rather than the possibility of altered toxicity results due to sperm-to-egg compatibility.

Acetylcholinesterase in the sea urchin Lytechinus variegatus: characterization and developmental expression in larvae.

Acetylcholinesterase (AChE) in the echinoid Lytechinus variegatus has been characterized. Kinetic parameters V(max), K(m), K(ss), and b were 2594+/-1048 nmol ATCh hydrolyzed/min/mg tissue wet weight, 185+/-11 microM, 308+/-100 mM, and 0.2, respectively for the substrate ATCh and 17.8+/-6.87 nmol BTCh hydrolyzed/min/mg tissue wet weight, 654+/-424 microM, 36+/-31 mM, and 0.6, respectively for BTCh. Pharmacologic analyses were performed with four inhibitors of cholinesterases, physostigmine, BW284c51, ethopropazine, and iso-OMPA, and yielded IC(50) values of 106+/-4 nM, 718+/-118 nM, 2.57+/-0.6 mM, and >0.0300 M, respectively. Both kinetic and pharmacologic results confirmed the existence of AChE in larval L. variegatus. Dimeric and tetrameric globular forms (determined by velocity sedimentation on sucrose gradients) were present in L. variegatus larvae. Activity of AChE increased significantly as larvae progressed in development from embryos to eight-arm larvae. Acetylcholinesterase activity of F1 larvae derived from sea urchins collected from wild populations and of F1 larvae derived from sea urchins cultured in the laboratory and fed two different diets suggest that the nutritional and/or environmental history of the adult sea urchin affect the developmental progression of AChE activity in the F1 offspring.

Regulation of spblimp1/krox1a, an alternatively transcribed isoform expressed in midgut and hindgut of the sea urchin gastrula.

The sea urchin regulatory gene Spblimp1/krox produces alternatively transcribed and spliced isoforms, 1a and 1b, which have different temporal and spatial patterns of expression. Here we describe a cis-regulatory module that controls the expression of the 1a splice form in the midgut and hindgut at the beginning of gastrulation. Conserved sequence patches revealed by a comparison of the blimp1/krox locus in Strongylocentrotus purpuratus and Lytechinus variegatus genomes were tested by gene transfer, in association with GFP or CAT reporter genes. An expression construct containing a conserved sequence patch immediately 5' of exon 1a included the transcription initiation site for blimp1/krox1a. This construct displays specific mid and hindgut expression, indicating that these are the locations of endogenous blimp1/krox1a transcription during the gastrula stage. Its sequence contains binding sites for Brn1/2/4, Otx, and Blimp1/Krox itself, as predicted in a prior regulatory network analysis.

A homologue of snail is expressed transiently in subsets of mesenchyme cells in the sea urchin embryo and is down-regulated in axis-deficient embryos.

Vertebrate members of the zinc finger transcription factor family related to Drosophila snail are expressed in neural crest and paraxial mesoderm along the left-right axis of the embryo. As simple deuterostomes, echinoderms are an important sister phylum for the chordates. We have identified populations of patterned, nonskeletogenic mesenchyme in the sea urchin Lytechinus variegatus by their expression of a sea urchin member of the snail family (Lv-snail). Lv-snail mRNA and protein are detectable at the midgastrula stage within the archenteron. At the late gastrula stage, a contiguous cluster of cells on the left side of the tip of the archenteron is Lv-snail-positive. At the early prism stage, two small clusters of mesenchyme cells near the presumptive arm buds are also Lv-snail-positive. At the pluteus stage, staining is detectable in isolated mesenchyme cells and the ciliated band. Based on fate mapping of secondary mesenchyme cells (SMCs) and double-label immunostaining, these patterns are consistent with expression of SNAIL by novel subsets of SMCs that are largely distinct from skeletogenic mesenchyme. In radialized embryos lacking normal bilateral symmetry, mesenchymal expression of Lv-SNAIL is abolished. These results suggest that transient expression of Lv-snail may be important for the differentiation of a subset of axially patterned nonskeletogenic mesenchyme cells and suggest conserved functions for snail family members in deuterostome development.

Ciclo reproductivo de Lytchinus variegatus (Echinoidea: Toxopfneustidae) en el sur de Isla Margarita, Venezuela / Reproductive cycle of Lytechinus variegatus (Echinoidea: Toxopneustidae) in the south of Margarita Island, Venezuela

The sea urchin Lytechinus variegatus is of some commercial importance on the Southern coast of Margarita Island (Venezuela), where artisan women fishers process and sell the roe. The reproduction cycle of this species had not been fully studied. We collected these sea urchins in two stations from November 1997 through October 1998. The samples were transported fresh to the laboratory to determine weight, diameter and height. The sexual stage was established with a macroscopic scale and the gonadosomatic index (GI). Maturity is achieved when the animals reaches a diameter of 22 mm, although 50% of the population reaches sexual development at diameters of 43 mm (males) and 45 mm (females) at Station 1, and 35 mm for both sexes at Station 2. Mature individuals, in a sexual proportion of 1:1 were collected every month. According to the monthly GI variation and frequency (%) of the development stages, on the southern beaches the main reproductive period begins in April and continues through October, and it is most intensive from May to July, when gonads reach the greatest growth

Desarrollo embrionario y larval de Lytechinus variegatus (Echinoidea: Toxopneustidae) en condiciones de laboratório en la Isla de Margarita-Venezuela / Embrionary and larval development of Lytechinus variegatus (Echinoidea: Toxopneustidae) in laboratory conditions at Isla de Margarita-Venezuela

The sea urchin Lytechinus variegatus is a promissory species for aquaculture activities in tropical countries. In Venezuela, this species has some economical importance but their embryonic and larval development had not been studied. We collected specimens from seagrass beds in Margarita Island (Venezuela) and kept them in the laboratory, where they spawned naturally. With filtered sea water (temperature 28 degrees C, salinity 37 psu) and moderate aeration, the eggs and sperm were mixed (relation 1:100) and reached a 90% fertilization rate. The fertilization envelope was observed after two minutes, the first cellular division after 45 minutes and the prism larval stage after 13 hours. The echinopluteus larval stage was reached after 17 hours and metamorphosis after 18 days of planktonic life, when the larvae start their benthic phase.

Producción de larvas y postlarvas del erizo verdiblanco del Caribe Lytechinus Variegatus (Echinodermata: Echinoidea) en condiciones de cultivo / 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

[Embrionary and larval development of Lytechinus variegatus (Echinoidea: Toxopneustidae) in laboratory conditions at Isla de Margarita-Venezuela]. / Desarrollo embrionario y larval de Lytechinus variegatus (Echinoidea: tOxopneustidae) en condiciones de laboratorio en la Isla de Margarita-Venezuela.

The sea urchin Lytechinus variegatus is a promissory species for aquaculture activities in tropical countries. In Venezuela, this species has some economical importance but their embryonic and larval development had not been studied. We collected specimens from seagrass beds in Margarita Island (Venezuela) and kept them in the laboratory, where they spawned naturally. With filtered sea water (temperature 28 degrees C, salinity 37 psu) and moderate aeration, the eggs and sperm were mixed (relation 1:100) and reached a 90% fertilization rate. The fertilization envelope was observed after two minutes, the first cellular division after 45 minutes and the prism larval stage after 13 hours. The echinopluteus larval stage was reached after 17 hours and metamorphosis after 18 days of planktonic life, when the larvae start their benthic phase.

[Reproductive cycle of Lytechinus variegatus (Echinoidea: Toxopneustidae) in the south of Margarita Island, Venezuela]. / Ciclo reproductivo de Lytechinus variegatus (Echinoidea: Toxopneustidae) en el sur de isla Margarita, Venezuela.

The sea urchin Lytechinus variegatus is of some commercial importance on the Southern coast of Margarita Island (Venezuela), where artisan women fishers process and sell the roe. The reproduction cycle of this species had not been fully studied. We collected these sea urchins in two stations from November 1997 through October 1998. The samples were transported fresh to the laboratory to determine weight, diameter and height. The sexual stage was established with a macroscopic scale and the gonadosomatic index (GI). Maturity is achieved when the animals reaches a diameter of 22 mm, although 50% of the population reaches sexual development at diameters of 43 mm (males) and 45 mm (females) at Station 1, and 35 mm for both sexes at Station 2. Mature individuals, in a sexual proportion of 1:1 were collected every month. According to the monthly GI variation and frequency (%) of the development stages, on the southern beaches the main reproductive period begins in April and continues through October, and it is most intensive from May to July, when gonads reach the greatest growth.

[Larvae and postlarvae production of the green-white sea urchin Lytechinus variegatus (Echinodermata: Echinoidea) in culture conditions]. / Producción de larvas y postlarvas del erizo verdiblanco del caribe Lytechinus variegatus (Echinodermata: Echinoidea) en condiciones de cultivo.

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.