Sea Urchin Research Articles

Investigating the timing of spawning of two sea urchins (Tripneustes gratilla and Toxopneustes pileolus)

AbstractMany marine organisms broadcast gametes for external fertilization, thus conspecifics need to concentrate the time of gamete release through synchronization with environmental factors. We investigated the timing of spawning in two sea urchin species, Tripneustes gratilla and Toxopneustes pileolus, by assessing the spawning potential of individuals using both surveys and experiments in southern Taiwan from March to November 2010. We developed a new method to observe the spawning potential without sacrificing any individuals, defining those individuals‐initiated spawning within 30 min after 10 s of exposure to air as having the spawning potential. We discovered that the percentage of individuals with spawning potential, that is, the spawning ratios of the population, varied greatly from 0% to 80% in To. pileolus and from 0% to 50% in Tr. gratilla within the reproductive seasons. We evaluated the possibility of a lunar/semilunar rhythm of spawning ratios of the population and tested three environmental factors, that is, light intensity, tide, and temperature that could potentially affect the spawning potential of the sea urchins by experiments. Individuals with spawning potential were neither randomly nor evenly distributed among days but were aggregated on certain days; however, no lunar/semilunar rhythm of the spawning ratios was detected within the reproductive season. The possible role of the tidal factor within a day was not found in experiments in either species. However, when water temperature was >27°C in our surveys, significantly higher spawning ratios were observed in To. pileolus, but not in Tr. gratilla in the reproductive season. A further temperature experiment in To. pileolus supported the positive role of high temperature on the spawning ratios of individuals. In addition, To. pileolus was more likely to spawn during the day than at night, and the light/dark experiments suggested that the spawning potential was controlled by an endogenous biological clock rather than by the ambient light conditions directly. The lack of rhythmic pattern and yet concentrated spawning of these sea urchins may be adaptive to avoid predation.

Characterization of thyrotropin-releasing hormone producing neurons in sea urchin, from larva to juvenile.

Most sea urchin species are indirect developers, going through a larval stage called pluteus. The pluteus possesses its own nervous system, consisting mainly of the apical organ neurons (controlling metamorphosis and settlement) and ciliary band neurons (controlling swimming behavior and food collection). Additional neurons are located in various areas of the gut. In recent years, the molecular complexity of this apparently "simple" nervous system has become apparent, with at least 12 neuronal populations identified through scRNA-sequencing in the species Strongylocentrotus purpuratus. Among these, there is a cluster of neurosecretory cells that produce a thyrotropin-releasing hormone-type neuropeptide (TRHergic) and that are also photosensory (expressing a Go-Opsin). However, much less is known about the organization of the nervous system in other sea urchin species. The aim of this work was to thoroughly characterize the localization of the TRHergic cells from early pluteus to juvenile stages in the Mediterranean sea urchin species Paracentrotus lividus combining immunostaining and whole mount in situ hybridization. We also compared the localization of TRHergic cells in early plutei of two other sea urchin species, Arbacia lixula and Heliocidaris tuberculata. This work provides new information on the anatomy and development of the nervous system in sea urchins. Moreover, by comparing the molecular signature of the TRHergic cells in P. lividus and S. purpuratus, we have obtained new insights how TRH-type neuropeptide signaling evolved in relatively closely related species.

Spontaneous intersibling polymorphism in the development of dopaminergic neuroendocrine cells in sea urchin larvae: impacts on the expansion of marine benthic species.

The plasticity of the nervous system plays a crucial role in shaping adaptive neural circuits and corresponding animal behaviors. Understanding the mechanisms underlying neural plasticity during development and its implications for animal adaptation constitutes an intriguing area of research. Sea urchin larvae offer a fascinating subject for investigation due to their remarkable evolutionary and ecological diversity, as well as their diverse developmental forms and behavioral patterns. We conducted immunochemical and histochemical analyses of serotonin-containing (5-HT-neurons) and dopamine-containing (DA-positive) neurons to study their developmental dynamics in two sea urchin species: Mesocentrotus nudus and Paracentrotus lividus. Our approach involved detailed visualization of 5-HT- and DA-positive neurons at gastrula-pluteus stages, coupled with behavioral assays to assess larval upward and downward swimming in the water column, with a focus on correlating cell numbers with larval swimming ability. The study reveals a heterochronic polymorphism in the appearance of post-oral DA-positive neuroendocrine cells and confirms the stable differentiation pattern of apical 5-HT neurons in larvae of both species. Notably, larvae of the same age exhibit a two- to four-fold difference in DA neurons. An increased number of DA neurons and application of dopamine positively correlate with larval downward swimming, whereas 5-HT-neurons and serotonin application induce upward swimming. The ratio of 5-HT/DA neurons determines the stage-dependent vertical distribution of larvae within the water column. Consequently, larvae from the same generation with a higher number of DA-positive neurons tend to remain at the bottom compared to those with fewer DA-positive neurons. The proportion of 5-HT and DA neurons within larvae of the same age underlies the different potentials of individuals for upward and downward swimming. A proposed model illustrates how coordination in humoral regulation, based on heterochrony in DA-positive neuroendocrine cell differentiation, influences larval behavior, mitigates competition between siblings, and ensures optimal population expansion. The study explores the evolutionary and ecological implications of these neuroendocrine adaptations in marine species.

Asymmetric segregation of maternal mRNAs and germline-related determinants in Cephalochordate embryos: implications for the evolution of early patterning events in chordates.

How animal embryos determine their early cell fates is an important question in developmental biology. In various model animals asymmetrically localized maternal transcripts play important roles in axial patterning and cell fate specification. Cephalochordates (amphioxus), which have three living genera (Asymmetron, Epigonichthys, Branchiostoma), are an early branching chordate lineage and thus occupy a key phylogenetic position for understanding the evolution of chordate developmental mechanisms. It has been shown that in the zygote of Brachiostoma amphioxus, which possess bilateral gonads flanking both sides of their trunk region, maternal transcripts of germline determinants form a compact granule. During early embryogenesis this granule is inherited by a single blastomere that subsequently gives rise to a cluster of cells displaying typical characteristics of primordial germ cells (PGC). These PGCs then come to lie in the tailbud region and proliferate during posterior elongation of the larva to join in the gonad anlagen at the ventral tip of the developing myomeres in amphioxus larvae. However, in Asymmetron and Epigonichthys amphioxus, whose gonads are present only on the right side of their body, nothing is known about their PGC development or the cellular/morphogenetic processes resulting in the asymmetric distribution of gonads. Using conserved germline determinants as markers, we show that similarly to Brachiostoma amphioxus, Asymmetron also employ a preformation mechanism to specify their PGCs, suggesting that this mechanism represents an ancient trait dating back to the common ancestor of Cephalochordates. Surprisingly, we found that Asymmetron PGCs are initially deposited on both sides of the body during early larval development; however, the left side PGCs cease to exist in young juveniles, suggesting that PGCs are eliminated from the left body side during larval development or following metamorphosis. This is reminiscent of the PGC development in the sea urchin embryo, and we discuss the implications of this observation for the evolution of developmental mechanisms.

Evolutionary and biogeographic patterns in the deep-sea echinoid families Pourtalesiidae Agassiz 1881 and Ceratophysidae fam. nov. (Echinoidea)

Abstract The sea urchin family Pourtalesiidae is primarily an abyssal taxon, exhibiting extremely modified morphologies that have emerged as an adaptation for burrowing in soft sediment. Here, we present the first detailed molecular phylogeny of the family Pourtalesiidae. Both morphological and molecular evidence support the establishment of the family Ceratophysidae fam. nov. to accommodate seven former pourtalesiid genera. The name Pourtalesiidae is retained for the group of genera Cystocrepis and Pourtalesia. Based on morphological data, the fossil genus Galeaster is transferred to the monotypic family Galeasteridae fam. nov.. Families Pourtalesiidae and Ceratophysidae are each characterized by a unique pattern of plastron plating. Reconstructions indicate that elongated tests, as well as other adaptations for burrowing, evolved independently in these families. The evolutionary histories of pourtalesiids and ceratophysids are complex and include several instances of increase and decrease in this specialization. Molecular data support the Antarctic origin of Pourtalesiidae and Ceratophysidae, although for the latter the support is limited. The initial dispersal of pourtalesiids from the Antarctic occurred in the northward direction and included distribution of the least specialized forms throughout the eastern Pacific. Broad radiation of the more specialized forms apparently occurred later. The biogeographic history of the family Ceratophysidae is more complex and at present cannot be reconstructed reliably.

Reproductive toxicity assessment of cellulose nanofibers, citric acid, and branched polyethylenimine in sea urchins: Eco-design of nanostructured cellulose sponge framework (Part B)

In the framework of a safe-by-design approach, we previously assessed the eco-safety of nanostructured cellulose sponge (CNS) leachate on sea urchin reproduction. It impaired gamete quality, gamete fertilization competence, and embryo development possibly due to the leaching of chemical additives used during the CNS synthesis process. To extend this observation and identify the component(s) that contribute to CNS ecotoxicity, in the present study, we individually screened the cytotoxic effects on sea urchin Arbacia lixula and Paracentrotus lividus gametes and embryos of the three main constituents of CNS, namely cellulose nanofibers, citric acid, and branched polyethylenimine. The study aimed to minimize any potential safety risk of these components and to obtain an eco-safe CNS. Among the three CNS constituents, branched polyethylenimine resulted in the most toxic agent. Indeed, it affected the physiology and fertilization competence of male and female gametes as well as embryo development in both sea urchin species. These results are consistent with those previously reported for CNS leachate. Moreover, the characterisation of CNS leachate confirmed the presence of detectable branched polyethylenimine in the conditioned seawater even though in a very limited amount. Altogether, these data indicate that the presence of branched polyethylenimine is a cause-effect associated with a significant risk in CNS formulations due to its leaching upon contact with seawater. Nevertheless, the suggested safety protocol consisting of consecutive leaching treatments and conditioning of CNS in seawater can successfully ameliorate the CNS ecotoxicity while maintaining the efficacy of its sorbent properties supporting potential environmental applications.

ROCK and the actomyosin network control biomineral growth and morphology during sea urchin skeletogenesis

Biomineralization had apparently evolved independently in different phyla, using distinct minerals, organic scaffolds, and gene regulatory networks (GRNs). However, diverse eukaryotes from unicellular organisms, through echinoderms to vertebrates, use the actomyosin network during biomineralization. Specifically, the actomyosin remodeling protein, Rho-associated coiled-coil kinase (ROCK) regulates cell differentiation and gene expression in vertebrates’ biomineralizing cells, yet, little is known on ROCK’s role in invertebrates’ biomineralization. Here, we reveal that ROCK controls the formation, growth, and morphology of the calcite spicules in the sea urchin larva. ROCK expression is elevated in the sea urchin skeletogenic cells downstream of the Vascular Endothelial Growth Factor (VEGF) signaling. ROCK inhibition leads to skeletal loss and disrupts skeletogenic gene expression. ROCK inhibition after spicule formation reduces the spicule elongation rate and induces ectopic spicule branching. Similar skeletogenic phenotypes are observed when ROCK is inhibited in a skeletogenic cell culture, indicating that these phenotypes are due to ROCK activity specifically in the skeletogenic cells. Reduced skeletal growth and enhanced branching are also observed under direct perturbations of the actomyosin network. We propose that ROCK and the actomyosin machinery were employed independently, downstream of distinct GRNs, to regulate biomineral growth and morphology in Eukaryotes.

ROCK and the actomyosin network control biomineral growth and morphology during sea urchin skeletogenesis

Biomineralization had apparently evolved independently in different phyla, using distinct minerals, organic scaffolds, and gene regulatory networks (GRNs). However, diverse eukaryotes from unicellular organisms, through echinoderms to vertebrates, use the actomyosin network during biomineralization. Specifically, the actomyosin remodeling protein, Rho-associated coiled-coil kinase (ROCK) regulates cell differentiation and gene expression in vertebrates’ biomineralizing cells, yet, little is known on ROCK’s role in invertebrates’ biomineralization. Here, we reveal that ROCK controls the formation, growth, and morphology of the calcite spicules in the sea urchin larva. ROCK expression is elevated in the sea urchin skeletogenic cells downstream of the Vascular Endothelial Growth Factor (VEGF) signaling. ROCK inhibition leads to skeletal loss and disrupts skeletogenic gene expression. ROCK inhibition after spicule formation reduces the spicule elongation rate and induces ectopic spicule branching. Similar skeletogenic phenotypes are observed when ROCK is inhibited in a skeletogenic cell culture, indicating that these phenotypes are due to ROCK activity specifically in the skeletogenic cells. Reduced skeletal growth and enhanced branching are also observed under direct perturbations of the actomyosin network. We propose that ROCK and the actomyosin machinery were employed independently, downstream of distinct GRNs, to regulate biomineral growth and morphology in Eukaryotes.

Local and regional variation in kelp loss and stability across coastal British Columbia

Kelp forests are among the most abundant coastal marine habitats but are vulnerable to climate change. The Northeast Pacific has experienced recent large-scale changes in kelp abundance and distribution, but little is known about changes north of the British Columbia (BC)-Washington border. Here, we assessed whether and how floating canopy kelp (Macrocystis pyrifera, Nereocystis luetkeana) distributions have changed in recent decades along the extensive coast of BC. We assembled and analysed available kelp distributional data, comparing snapshots of kelp linear extent from 1.5-3 decades ago (1994-2007) to recently collected data (2017-2021) across 11 different subregions spanning the province. We then leveraged timeseries, where available (n = 7 data sets), to contextualise patterns of change. In aggregate, the data suggest that kelp forests have declined considerably in some parts of the province, but with variable patterns of change across BC. In the warmest areas (southern BC), kelp persistence was negatively correlated with mean summer sea surface temperatures, which at times exceeded known thermal tolerances. In contrast, in northern subregions, top-down control by sea urchins and otters appeared to modulate kelp dynamics, with declines occurring in 2 subregions despite cool ocean temperatures. Timeseries data suggest that many declines occurred around the 2014-2016 marine heatwave, an event associated with sustained warming and altered trophic dynamics. Our results suggest that the extent of BC’s kelp forests has declined in some places in recent decades, but that regional and local-scale factors influence their responses to environmental change.

Sea Urchin Food Waste into Bioactives: Collagen and Polyhydroxynaphtoquinones from P. lividus and S. granularis

Approximately 75,000 tons of different sea urchin species are globally harvested for their edible gonads. Applying a circular economy approach, we have recently demonstrated that non-edible parts of the Mediterranean Sea urchin Paracentrotus lividus can be fully valorized into high-value products: antioxidant pigments (polyhydroxynaphthoquinones—PHNQs) and fibrillar collagen can be extracted to produce innovative biomaterials for biomedical applications. Can waste from other edible sea urchin species (e.g., Sphaerechinus granularis) be similarly valorised? A comparative study on PHNQs and collagen extraction was conducted. PHNQ extraction yields were compared, pigments were quantified and identified, and antioxidant activities were assessed (by ABTS assay) and correlated to specific PHNQ presence (i.e., spinochrome E). Similarly, collagen extraction yields were evaluated, and the resulting collagen-based biomaterials were compared in terms of their ultrastructure, degradation kinetics, and resistance to compression. Results showed a partially similar PHNQ profile in both species, with significantly higher yield in P. lividus, while S. granularis exhibited better antioxidant activity. P. lividus samples showed higher collagen extraction yield, but S. granularis scaffolds showed higher stability. In conclusion, waste from different species can be successfully valorised through PHNQ and collagen extraction, offering diverse applications in the biomedical field, according to specific technical requirements.

Multi‐omic analysis reveals genes and proteins integral to bioactivity of Echinochrome A isolated from the waste stream of the sea urchin industry in Aotearoa New Zealand

AbstractEvechinus chloroticus (commonly known as kina) is a sea urchin species endemic to New Zealand. Its roe is a culinary delicacy to the indigenous Māori and a globally exported food product. Echinochrome A (Ech A) is a bioactive compound isolated from the waste product of kina shells and spines; however, the molecular mechanisms of Ech A bioactivity are not well understood, partly due to Ech A never being studied using unbiased genome‐wide analysis. To explore the high‐value pharmaceutical potential of kina food waste, we obtained unbiased functional genomic and proteomic profiles of yeast cells treated with Echinochrome A. Abundance was measured for 4100 proteins every 30 min for four hours using fluorescent microscopy, resulting in the identification of 92 proteins with significant alterations in protein abundance caused by Ech A treatment that were over‐represented with specific changes in DNA replication, repair and RNA binding after 30 min, followed by specific changes in the metabolism of metal ions (specifically iron and copper) from 60–240 min. Further analysis indicated that Ech A chelated iron, and that iron supplementation negated the growth inhibition caused by Ech A. Via a growth‐based genome‐wide analysis of 4800 gene deletion strains, 20 gene deletion strains were sensitive to Ech A in an iron‐dependent manner. These genes were over‐represented in the cellular response to oxidative stress, suggesting that Ech A suppressed growth inhibition caused by oxidative stress. Unexpectedly, genes integral to cardiolipin and inositol phosphate biosynthesis were required for Ech A bioactivity. Overall, these results identify genes, proteins, and cellular processes mediating the bioactivity of Ech A. Moreover, we demonstrate unbiased genomic and proteomic methodology that will be useful for characterizing bioactive compounds in food and food waste.

Interactive effects of chronic ocean acidification and warming on the growth, survival, and physiological responses of adults of the temperate sea urchin Strongylocentrotusintermedius

To investigate the interactive effects of chronic ocean acidification and warming (OAW) on the growth, survival, and physiological responses of sea urchins, adults of the temperate sea urchin Strongylocentrotus intermedius were incubated separately/jointly in acidic (ΔpHNBS = −0.5 units) and thermal (ΔT = +3.0 °C) seawater for 120 days under lab-controlled conditions based on the projected ocean pH and temperature for 2100 put forward by the Intergovernmental Panel on Climate Change (IPCC). Survival rate (SR), average food consumption rate (FCR), gut index (GuI), specific growth rate (SGR), digestive capability, energy production, and antioxidant capability were subsequently determined. The results showed that 1) the SR, FCR, GuI and SGR decreased sharply under OAW conditions. Significant interactive effects of OAW on SR and SGR were observed at 120 days post-incubation (dpi), and on FCR this occurred at 90 dpi. 2) OAW altered the activities of both digestive and antioxidant enzymes. There were significant interaction effects of OAW on the activities of amylase, trehalase, and superoxide dismutase. 3) The relative gene expression levels and activities of key enzymes involved in glycometabolism pathways (i.e., glycolysis and the tricarboxylic acid cycle) were significantly affected by OAW, resulting in an alteration of the total ATP content in the sea urchins. Interaction effects of OAW were observed in both relative gene expression and the activity of enzymes involved in glycolysis (hexokinase), the transformation of glycolysis end-products (lactate dehydrogenase), the tricarboxylic acid cycle (citrate synthetase), and ATP production (Na+/K+-ATPase). The data from this study will enrich our knowledge concerning the combined effects of global climate change on the survival, growth, and physiological responses of echinoderms.

Genomic signatures of exceptional longevity and negligible aging in the long-lived red sea urchin

The red sea urchin (Mesocentrotus franciscanus) is one of the Earth's longest-living animals, reported to live more than 100 years with indeterminate growth, life-long reproduction, and no increase in mortality rate with age. To understand the genetic underpinnings of longevity and negligible aging, we constructed a chromosome-level assembly of the red sea urchin genome and compared it to that of short-lived sea urchin species. Genome-wide syntenic alignments identified chromosome rearrangements that distinguish short- and long-lived species. Expanded gene families in long-lived species play a role in innate immunity, sensory nervous system, and genome stability. An integrated network of genes under positive selection in the red sea urchin was involved in genomic regulation, mRNA fidelity, protein homeostasis, and mitochondrial function. Our results implicated known longevity genes in sea urchin longevity but also revealed distinct molecular signaturesthat may promote long-term maintenance of tissue homeostasis, disease resistance, and negligible aging.

Testing sea urchin and green sea turtle consumption of the allelopathic macroalga Galaxaura divaricata.

Galaxaura divaricata is a partially calcified macroalga that hampers coral recruitment, growth, and recovery via the excretion of allelopathic secondary metabolites. Herbivorous fishes are not major consumers of Galaxaura spp. and there is a need to understand feeding preferences for Galaxaura divaricata in other macroherbivores, like sea urchins and green sea turtles that could act as potential controlling agents. Under certain environmental conditions, G. divaricata can proliferate and overgrow degraded reefs for several years, as documented for several coral patch reefs in the lagoon of Dongsha Atoll, South China Sea. This study aimed to experimentally test the feeding preferences of five species of sea urchin and two individual green sea turtles, Chelonia mydas, for G. divaricata. Specifically, we quantified and compared the consumption rates of the allelopathic G. divaricata with Gracilaria edulis, a nonallelopathic, fleshy red alga, known to be highly favored by herbivores. Results showed that the five urchin species fed on both G. edulis and G. divaricata. However, urchins consumed 2-8 times less wet weight of G. divaricata (range 0.3-3.1 g urchin-1 24 h-1) compared to G. edulis (range 0.6-18 g urchin-1 24 h-1), suggesting that urchin grazing may exert some control on G. divaricata abundance but is likely ineffective for a large-scale removal of the alga. Further, both green sea turtles avoided G. divaricata and selectively fed on G. edulis. More experiments are needed to test the potential role of herbivores in controlling the overgrowth of coral competitive and allelopathic macroalgae, like Galaxaura on coral reefs.

Marine sulfated glycans inhibit the interaction of heparin with S-protein of SARS-CoV-2 Omicron XBB variant.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a worldwide COVID-19 pandemic, leading to 6.8million deaths. Numerous variants have emerged since its outbreak, resulting in its significantly enhanced ability to spread among humans. As with many other viruses, SARS‑CoV‑2 utilizes heparan sulfate (HS) glycosaminoglycan (GAG) on the surface of host cells to facilitate viral attachment and initiate cellular entry through the ACE2 receptor. Therefore, interfering with virion-HS interactions represents a promising target to develop broad-spectrum antiviral therapeutics. Sulfated glycans derived from marine organisms have been proven to be exceptional reservoirs of naturally existing HS mimetics, which exhibit remarkable therapeutic properties encompassing antiviral/microbial, antitumor, anticoagulant, and anti-inflammatory activities. In the current study, the interactions between the receptor-binding domain (RBD) of S-protein of SARS-CoV-2 (both WT and XBB.1.5 variants) and heparin were applied to assess the inhibitory activity of 10 marine-sourced glycans including three sulfated fucans, three fucosylated chondroitin sulfates and two fucoidans derived from sea cucumbers, sea urchin and seaweed Saccharina japonica, respectively. The inhibitory activity of these marine derived sulfated glycans on the interactions between RBD of S-protein and heparin was evaluated using Surface Plasmon Resonance (SPR). The RBDs of S-proteins from both Omicrion XBB.1.5 and wild-type (WT) were found to bind to heparin, which is a highly sulfated form of HS. All the tested marine-sourced sulfated glycans exhibited strong inhibition of WT and XBB.1.5S-protein binding to heparin. We believe the study on the molecular interactions between S-proteins and host cell glycosaminoglycans provides valuable insight for the development of marine-sourced, glycan-based inhibitors as potential anti-SARS-CoV-2 agents.

Transgenerational acclimation to acidified seawater and gene expression patterns in a sea urchin

Transgenerational responses of susceptible calcifying organisms to progressive ocean acidification are an important issue in reducing uncertainty of future predictions. In this study, a two-generation rearing experiment was conducted using mature Mesocentrotus nudus, a major edible sea urchin that occurs along the coasts of northern Japan. Morphological observations and comprehensive gene expression analysis (RNA-seq) of resulting larvae were performed to examine transgenerational acclimation to acidified seawater. Two generations of rearing experiments showed that larvae derived from parents acclimated to acidified seawater tended to have higher survival and show less reduction in body size when exposed to acidified seawater of the same pH, suggesting that a positive carry-over effect occurred. RNA-seq analysis showed that gene expression patterns of larvae originated from both acclimated and non-acclimated parents to acidified seawater tended to be different than control condition, and the gene expression pattern of larvae originated from acclimated parents was substantially different than that of larvae of non-acclimated and control parents.

Developmental and Biochemical Markers of the Impact of Pollutant Mixtures Under the Effect of Global Climate Change

This study investigates the combined impact of microplastics (MP) and Chlorpyriphos (CPF) on sea urchin larvae (Paracentrotus lividus) under the backdrop of ocean warming and acidification. While the individual toxic effects of these pollutants have been previously reported, their combined effects remain poorly understood. Two experiments were conducted using different concentrations of CPF (EC10 and EC50) based on previous studies from our group. MP were adsorbed in CPF to simulate realistic environmental conditions. Additionally, water acidification and warming protocols were implemented to mimic future ocean conditions. Sea urchin embryo toxicity tests were conducted to assess larval development under various treatment combinations of CPF, MP, ocean acidification (OA), and temperature (OW). Morphometric measurements and biochemical analyses were performed to evaluate the effects comprehensively. Results indicate that combined stressors lead to significant morphological alterations, such as increased larval width and reduced stomach volume. Furthermore, biochemical biomarkers like acetylcholinesterase (AChE), glutathione S-transferase (GST), and glutathione reductase (GRx) activities were affected, indicating oxidative stress and impaired detoxification capacity. Interestingly, while temperature increase was expected to enhance larval growth, it instead induced thermal stress, resulting in lower growth rates. This underscores the importance of considering multiple stressors in ecological assessments. Biochemical biomarkers provided early indications of stress responses, complementing traditional growth measurements. The study highlights the necessity of holistic approaches when assessing environmental impacts on marine ecosystems. Understanding interactions between pollutants and environmental stressors is crucial for effective conservation strategies. Future research should delve deeper into the impacts at lower biological levels and explore adaptive mechanisms in marine organisms facing multiple stressors. By doing so, we can better anticipate and mitigate the adverse effects of anthropogenic pollutants on marine biodiversity and ecosystem health.

Fabrication of magnesium-doped biphasic calcium phosphate granules with sea urchin spine-derived porous structure

In bone regeneration, synthetic materials are required to replace autogenous bone grafts. The shape, composition, surface, and porous structure of the material are key factors in bone formation and material resorption. In this study, we describe the fabrication of Mg-doped biphasic calcium phosphate (Mg-BCP) granules comprising hydroxyapatite (HAp) and beta-tricalcium phosphate (β-TCP) by exploiting the structural and chemical characteristics of sea urchin spines. The Mg-BCP granules were cylinders with a 1.2–1.8 mm diameter and height of 1.4–1.7 mm. The β-TCP/HAp ratio and Mg content in the Mg-BCP granules were controlled within the ranges of 0.36–0.45 and 2.40–6.21 mol%, respectively. Controlling the β-TCP/HAp ratio and Mg content may contribute to controlling the material resorption rate and promoting osteogenesis and angiogenesis. The Mg-BCP granules maintained the structural characteristics of the sea urchin spines. The granules comprised radial wedges termed septa, and a meshwork termed stereom. The interseptal gaps, or micropores, decreased from the granular exterior (24–29 μm) to the interior (14–28 μm). The septa in the outermost layer provide concavities on the sides of the cylindrical granules. The stereom meshworks in the core and outer layers involved micropores in the ranges of 3–15 and 3–10 μm, respectively. These multiscale pores and surface undulations may improve cell penetration and attachment, bodily fluid permeability, and protein adsorption. The compressive strengths of the Mg-BCP granules were in the range of 45.2–46.1 MPa, which are useful as bone grafts. Due to their rounded shapes, the Mg-BCP granules filled a mold more densely than the clinically used and irregularly shaped granules. These findings demonstrated that Mg-BCP granules are promising materials for bone regeneration.

Phylogenetic Relationship of Diadema: Emphasis on The Two Distinct Clades of D. Setosum With The Inclusion of Long Spine Black Sea Urchin From Malaysian Borneo

Diadema urchins (family Diadematidae) are ecologically important bioindicators of coral reef ecosystems and seagrass beds. Diadema urchins which are widely distributed and broadcast spawners, have been frequently utilized as model invertebrate species for zoogeography research of the Indo-West Pacific region. So far, Malaysian Borneo, located at the geographic center of Maritime Southeast Asia, has been under-sampled. This study aims to fill this sampling gap and provide the first record of Diadema setosum from Malaysian Borneo using genetic diagnostics to conclusively establish the clade-level identity of the species. According to Cytochrome Oxidase I gene analysis, Diadema is monophyletic. Seven species of Diadema namely Diadema palmeri, Diadema clarki, Diadema mexicanum, Diadema antillarum, Diadema paucispinum, Diadema africanum, and Diadema savignyi, formed their subclades with strong bootstrap values, demonstrating interspecific variation. The findings of this study provide further evidence for the presence of two distinct monophyletic clades, with all D. setosum individuals forming a monophyletic clade that later split into two distinct subclades, dividing Red Sea population (D. setosum-b) and Indo-West Pacific populations (D. setosum-a), supported by a significant genetic divergence value ranging from 6.3% to 9.1%. This study also revealed notable levels of nucleotide and population subdivision between the D. setosum from the Indo-West Pacific and the Red Sea populations (Nst = 0.891; Fst = 0.886) with a low number of migrants per generation (Nm = 0.065). This may suggest geographic isolation due to ecological factors preventing each other from surviving in the territory of the other, or that the two clades of D. setosum were a separate species. Additional morphological and molecular analysis is required in the future to ascertain the level of divergence and further resolve the taxonomic confusion within the genus Diadema.

Determination of Micro Minerals of Several Species of Sea Urchins from Samboang Waters as Functional Food Candidates

Functional foods are foods that can be consumed with additional health benefits beyond their basic function as an energy source. The search for food sources from the sea is increasing, with many discoveries of active compounds from marine organisms. This research aims to determine the micromineral content of iron (Fe), zinc (Zn), and iodine (I) in several species of sea urchins in the coastal waters of Samboang Bulukumba. The results of this research can be a basis for utilizing this biota as a functional food. This laboratory research uses micromineral measurement instruments, for example, Atomic Absorption Spectrophotometry (AAS) for Fe and Zn, while for iodine minerals using High Performed Liquid Chromatography (HPLC) instruments. In this study, 2 types of sea urchins were found which were identified as Diadema cytosum and Tripneustes gratilla. The results of measuring the water content and ash content of the gonads of the two sea urchins were 77.32% and 1.15% for Diadema cytosum and 72.22% and 2.09% for Tripneustes gratilla. The results of measuring the levels of micro minerals Fe, Zn, and Iodine were respectively 115.24 ppm, 31.44 ppm and 16.71 ppm for Diadema cytosum and 150.75 and 27.27 ppm 21.21 ppm for Tripneustes gratilla. Keywords: functional food, AAS, HPLC, Diadema cytosum, Tripneustes gratilla Abstrak Pangan fungsional adalah makanan yang dapat dikonsumsi dengan manfaat kesehatan tambahan di luar dari fungsi dasarnya sebagai sumber energi. Pencarian sumber pangan dari laut semakin meningkat, dengan banyaknya penemuan senyawa-senyawa aktif dari organisme laut. Penelitian ini bertujuan  menentukan kandungan mikro  mineral besi (Fe), seng (Zn) dan yodium (I) dalam beberapa spesies bulu babi di perairan pantai Samboang Bulukumba. Hasil penelitian ini dapat menjadi dasar dalam memanfaatkan biota tersebut sebagai pangan fungsional. Penelitian laboratorium ini menggunakan instrumen pengukuran mineral mikro misalnya dengan  penggunaan Atomic Absorption Spectrofotometry (AAS) untuk Fe dan Zn, sedangkan untuk mineral yodium menggunakan instrumen High Performed Liquid Chromatography (HPLC). Dalam penelitian ini ditemukan 2 jenis bulu babi yang teridentifikasi sebagai  Diadema sitosum dan Tripneustes gratilla. Hasil pengukuran kadar air dan kadar abu gonad kedua bulu babi tersebut  diperoleh 77.32% dan 1.15% untuk Diadema sitosum dan sebesar 72.22% dan 2.09% untuk Tripneustes gratilla.  Hasil pengukuran  kadar mineral mikro Fe,  Zn dan Yodium masing-masing adalah sebesar 115.24  ppm, 31.44 ppm dan 16.71ppm untuk Diadema sitosum dan 150.75 dan 27.27 ppm 21.21 ppm untuk Tripneustes gratilla. Kata kunci : pangan fungsional, AAS, HPLC, Diadema sitosum, Tripneustes gratilla