Establishment of Parentage Identification Method for Sea Urchin Strongylocentrotus intermedius Based on SSR-seq Technology.

To establish a parentage identification method for Strongylocentrotus intermedius, 15 microsatellite loci and simple sequence repeat sequencing (SSR-seq) technology were used to perform SSR sequencing and typing of the validation population with known pedigree information and the simulation population. Cervus v3.0 was used for gene frequency statistics, simulated analysis, and parentage identification analysis. The results showed that, in validation population, using 15 microsatellite loci, the highest success rate of parent pairs identification was 86%, the highest success rate of female parent identification was 93%, and the highest success rate of male parent identification was 90%. The simulated population was analyzed using 12-15 loci, and the identification rate was up to 90%. In cases where accurate parentage was not achieved, individuals could exhibit genetic similarities with 1-3 male or female parents. Individuals identified as lacking a genetic relationship can be selected as parents to prevent inbreeding. This study shows that parent pairs or single parents of most offspring can be identified successfully using these 15 selected loci. The results lay a foundation for the establishment of a parentage identification method for S. intermedius.

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.

Hidden in plain sight: hyperspectral documentation of complex biofluorescence produced by the green sea urchin (Strongylocentrotus droebachiensis).

Biofluorescence in echinoderms is largely unexplored, and even though the green sea urchinStrongylocentrotus droebachiensisis a well-studied species, the presence and/or function of fluorescence remains very poorly understood. Hyperspectral imaging was conducted on adult sea urchins (N = 380) while fluorospectrometric analysis was conducted on sea urchin coelomic fluid (N = 30). Fluorescence was documented in both the spines and coelomic fluid ofS. droebachiensis. Intact spines exhibited a low intensity green emission (∼550-600 nm), while broken spines averaged a high emission peak in the green spectrum (∼580 nm). Sea urchins produce a red exudate with a pronounced emission peak (∼680 nm) with a shoulder peak (∼730 nm). The sampled coelomic fluid exhibited high variability, with a majority exhibiting a low-level green fluorescence while pronounced emission peaks (N = 5) were found in the red spectrum (∼680 nm). The complex fluorescence produced byS. droebachiensiswarrants further investigation on its applicability for monitoring welfare of sea urchins in aquaculture facilities.

TMT-based proteomics analysis of sea urchin (Strongylocentrotus intermedius) under high temperature stress.

In the context of global warming and continuous high temperatures in the northern part of China in summer, the mortality rate of Strongylocentrotus intermedius through the summer reaches 70-80 %. The protein regulatory mechanism of S. intermedius in response to high temperature stress is still unclear. In order to investigate the protein expression of S. intermedius under high temperature stress, the study was conducted with the high-temperature resistant strain of S. intermedius and the control group of S. intermedius. Tandem Mass Tag (TMT) tagging technique was applied to resolve the protein expression profile of S. intermedius in response to high temperature stress. The results showed that, compared to 15 °C，136 DEPs were screened in high-temperature resistant strain groups of S. intermedius under high temperature stress and 87 DEPs were screened in the control group of S. intermedius. There were 33 common differential proteins in the two groups, such as APOLP, HSP 70, CDC37 and CALM. Further Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyses revealed that the up-regulated proteins CALM and HSP70 are significantly enriched in the "Phosphatidylinositol signaling system" and "Protein processing in endoplasmic reticulum" in heat-tolerant S. intermedius strains under high temperature stress. The control group of S. intermedius DEPs were significantly enriched in protein processing in the endoplasmic reticulum. These results provide a theoretical basis for the molecular mechanism of sea urchin heat tolerance and fundamental data for sea urchin selection and breeding for high temperature tolerance.

High temperature influences DNA methylation and transcriptional profiles in sea urchins (Strongylocentrotus intermedius).

BACKGROUND: DNA methylation plays an important role in life processes by affecting gene expression, but it is still unclear how DNA methylation is controlled and how it regulates gene transcription under high temperature stress conditions in Strongylocentrotus intermedius. The potential link between DNA methylation variation and gene expression changes in response to heat stress in S. intermedius was investigated by MethylRAD-seq and RNA-seq analysis. We screened DNA methylation driver genes in order to comprehensively elucidate the regulatory mechanism of its high temperature adaptation at the DNA/RNA level. RESULTS: The results revealed that high temperature stress significantly affected not only the DNA methylation and transcriptome levels of S. intermedius (P < 0.05), but also growth. MethylRAD-seq analysis revealed 12,129 CG differential methylation sites and 966 CWG differential methylation sites, and identified a total of 189 differentially CG methylated genes and 148 differentially CWG methylated genes. Based on KEGG enrichment analysis, differentially expressed genes (DEGs) are mostly enriched in energy and cell division, immune, and neurological damage pathways. Further RNA-seq analysis identified a total of 1968 DEGs, of which 813 genes were upregulated and 1155 genes were downregulated. Based on the joint MethylRAD-seq and RNA-seq analysis, metabolic processes such as glycosaminoglycan degradation, oxidative phosphorylation, apoptosis, glutathione metabolism, thermogenesis, and lysosomes are regulated by DNA methylation. CONCLUSIONS: High temperature affected the DNA methylation and expression levels of genes such as MOAP-1, GGT1 and RDH8, which in turn affects the metabolism of HPSE, Cox, glutathione, and retinol, thereby suppressing the immune, energy metabolism, and antioxidant functions of the organism and finally manifesting as stunted growth. In summary, the observations in the present study improve our understanding of the molecular mechanism of the response to high temperature stress in sea urchin.

Strongylocentrotus nudus egg polysaccharide (SEP) suppresses HBV replication via activation of TLR4-induced immune pathway.

Chronic hepatitis B virus (HBV) infection is a worldwide public health problem that causes significant liver-related morbidity and mortality. In our previous study, Strongylocentrotus nudus eggs polysaccharide (SEP), extracted from sea urchins, had immunomodulatory and antitumor effects. Whether SEP has anti-HBV activity is still obscure. This study demonstrated that SEP decreased the secretion of hepatitis B surface antigen (HBsAg) and e antigen (HBeAg), as well as the replication and transcription of HBV both in vitro and in vivo. Immunofluorescence and immunohistochemistry results showed that the level of HBV core antigen (HBcAg) was clearly reduced by SEP treatment. Mechanistically, RT-qPCR, western blot, and confocal microscopy analysis showed that SEP significantly increased the expression of toll-like receptor 4 (TLR4) and co-localization with TLR4. The downstream molecules of TLR4, including NF-κb and IRF3, were activated and the expression of IFN-ß, TNF-α, IL-6, OAS, and MxA were also increased, which could suppress HBV replication. Moreover, SEP inhibited other genotypes of HBV and hepatitis C virus (HCV) replication in vitro. In summary, SEP could be investigated as a potential anti-HBV drug capable of modulating the innate immune.

Polysaccharide from Strongylocentrotus nudus eggs regulates intestinal epithelial autophagy through CD36/PI3K-Akt pathway to ameliorate inflammatory bowel disease.

Sea urchin is a popular food all over the world, of which eggs are main edible part. Previous studies suggested that polysaccharides from eggs of Strongylocentrotus nudus (SEP) exhibited immunomodulatory activities during anti-tumor therapy, nevertheless, effects of SEP on inflammatory bowel disease and its underlying mechanisms have never been reported. In the present study, we showed that the SEP inhibited dextran sodium sulfate-induced ulcerative colitis characterized by decreased disease activity index, restored colon length and body weight, improved histopathological changes, down-regulation of inflammatory cytokines levels and Th17/Treg ratios in C57BL/6 J mice. Moreover, immunofluorescence analysis suggested that SEP repaired gut barrier in UC mice, while 16S rDNA sequencing exhibited improved intestinal flora. Mechanistically, we found SEP significantly modulated autophagy-related factors in intestinal epithelial cells (IECs), while might contributed to pathogenesis of UC. Furthermore, we demonstrated PI3K/Akt pathway was involved in regulatory effect of SEP on lipopolysaccharide-induced autophagy of HT-29 cells. Besides, among possible polysaccharide binding receptors, change of the CD36 expression was most significant, which was associated with PI3K/Akt signals. Collectively, our study showed for the first time that the SEP might be used a prebiotic agent to improve IBD through regulating CD36-PI3K/Akt mediated autophagy of IECs.

Integrative mRNA-miRNA interaction analysis associated with the immune response of Strongylocentrotus intermedius to Vibrio harveyi infection.

Strongylocentrotus intermedius is one of the most economically valuable sea urchin species in China and has experienced mass mortality owing to outbreaks of bacterial diseases such as black mouth disease. This has caused serious economic losses to the sea urchin farming industry. To investigate the immune response mechanism of S. intermedius with different tube feet colors in response to Vibrio harveyi infection, we examined the different tube feet-colored S. intermedius under V. harveyi challenge and compared their transcriptome and microRNA (miRNA) profiles using RNA-Seq. We obtained 1813 differentially expressed genes (DEGs), 28 DE miRNAs, and 303 DE miRNA-DEG pairs in different tube feet-colored S. intermedius under V. harveyi challenge. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the most significant DEGs were associated with the Notch signaling and phagosome pathways. The target genes of immune-related miRNAs (miR-71, miR-184, miR-193) and genes (CALM1, SPSB4, DMBT, CSRP1) in S. intermedius were predicted and validated. This study provides insight into the molecular mechanisms that regulate genes involved in the immune response of S. intermedius infected with V. harveyi.

Nanoplastics affect the growth of sea urchins (Strongylocentrotus intermedius) and damage gut health.

Nanoplastics (NPs) are abundant and widespread throughout the ocean, not only causing severe environmental pollution, but also worsening the aquatic organisms. To elucidate the mechanism of biological toxic effects underlying the responses of marine invertebrates to NPs, Strongylocentrotus intermedius was stressed with three different NPs concentrations (0 particles/L, 102 particles/L and 104 particles/L). Specific growth rates, enzyme activity, gut tissue section observation and structural characteristics of the gut bacterial community were analyzed. After 28 days of exposure, the specific growth rate of S. intermedius decreased significantly with NPs groups. Further, both lysozyme, pepsin, lipase and amylase activities decreased, while the superoxide dismutase activity increased, indicating that NPs negatively affected digestive enzyme and immune enzyme activity. The analysis of gut tissue sections revealed that NPs caused atrophy and cytoplasmic reduction in the epithelial cells of the S. intermedius intestine. Moreover, the structural characterization of the gut bacterial community indicated significant changes in the abundances of members from Campylobacterota, Chlamydiae, and Firmicutes. Members from Arcobacteraceae, Christensenellaceae and Clostridia were endemic to the NPs treatment. The KEGG database analysis demonstrated that the metabolic pathways specific to the NPs treatment group were significantly associated with growth, energy metabolism, and immunity. In summary, NPs have negatively affected on physiological response and altered gut microecological environment.

Quinoid Pigments of Sea Urchins Scaphechinus mirabilis and Strongylocentrotus intermedius: Biological Activity and Potential Applications.

This review presents literature data: the history of the discovery of quinoid compounds, their biosynthesis and biological activity. Special attention is paid to the description of the quinoid pigments of the sea urchins Scaphechinus mirabilis (from the family Scutellidae) and Strongylocentrotus intermedius (from the family Strongylocentrotidae). The marine environment is considered one of the most important sources of natural bioactive compounds with extremely rich biodiversity. Primary- and some secondary-mouthed animals contain very high concentrations of new biologically active substances, many of which are of significant potential interest for medical purposes. The quinone pigments are products of the secondary metabolism of marine animals, can have complex structures and become the basis for the development of new natural products in echinoids that are modulators of chemical interactions and possible active ingredients in medicinal preparations. More than 5000 chemical compounds with high pharmacological potential have been isolated and described from marine organisms. There are three well known ways of naphthoquinone biosynthesis-polyketide, shikimate and mevalonate. The polyketide pathway is the biosynthesis pathway of various quinones. The shikimate pathway is the main pathway in the biosynthesis of naphthoquinones. It should be noted that all quinoid compounds in plants and animals can be synthesized by various ways of biosynthesis.

Trans-Arctic vicariance in Strongylocentrotus sea urchins.

The sea urchins Strongylocentotus pallidus and S. droebachiensis first invaded the Atlantic Ocean from the Pacific following the opening of the Bering seaway in the late Miocene. While trans-Arctic dispersal during the Pleistocene is thought to have maintained species' integrity, a recent genomic analysis identified a reproductively isolated cryptic species within S. droebachiensis. Based on previous studies, the distribution of one of these lineages (S. droebachiensis W) includes the shallow water habitats of the northwest Atlantic and Pacific, while the other (S. droebachiensis E) is found throughout the shallow habitat in the northeast but is mostly restricted to deep habitats (>65 m) in the northwest Atlantic. However, since genetic variation within S. droebachiensis has been largely unstudied in the north Pacific and Arctic oceans, the biogeography of the cryptic species is not well known, and it is difficult to identify the mechanisms driving population subdivision and speciation. Here we use population genetic analyses to characterize the distribution of each species, and to test hypotheses about the role of vicariance in the evolution of systematic and genomic divergence within the genus. We collected individuals of all three Strongylocentrotus species (n = 365) from 10 previously unsampled locations in the northeast Pacific and north Atlantic (Labrador Sea and Norway), and generated mtDNA sequence data for a 418 bp fragment of cytochrome c oxidase subunit I (COI). To assess the biogeography of all three species, we combined our alignment with five previously published data sets (total n = 789) and used statistical parsimony and maximum likelihood to identify species and characterize their distribution within and among oceans. Patterns of haplotype sharing, pairwise F ST , and hierarchical analyses of molecular variance (AMOVA) identified trans-Arctic dispersal in S. pallidus and S. droebachiensis W, but other than 5 previously reported singletons we failed to detect additional mtDNA haplotypes of S. droebachiensis E in the north Pacific. Within the Atlantic, patterns of habitat segregation suggests that temperature may play a role in limiting the distribution of S. droebachiensis E, particularly throughout the warmer coastal waters along the coast of Nova Scotia. Our results are consistent with the cycles of trans-Arctic dispersal and vicariance in S. pallidus and S. droebachiensis W, but we suggest that the evolution of Atlantic populations of S. droebachiensis E has been driven by persistent trans-Arctic vicariance that may date to the initial invasion in the late Pliocene.

Ecological niche models for the assessment of site suitability of sea cucumbers and sea urchins in China.

In the present study, the maximum entropy model (MaxEnt) based on the data of sea surface temperature (SST) and published information was used to assess the site suitability for the aquaculture expansion of the sea cucumber Apostichopus japonicus and the sea urchin Strongylocentrotus intermedius in China. According to the current assessment, the coastal areas of Hebei province and Tianjin have great prospects for A. japonicus aquaculture, while is currently being underutilized. In the south, more than 94% of the coastal areas in Zhejiang, Fujian, and Guangdong provinces are suitable for the growth of A. japonicus for six months, especially the coastal areas of Lianjiang, Changle, Fuqing and Putian in Fujian province. The water temperatures in more than 94% of China's coastal areas are higher than 25 °C in July and August, which probably results in the mortality of S. intermedius in aquaculture. This clearly indicates that high water temperature is the bottleneck of S. intermedius aquaculture and well explains the limited expansion of this commercially important exotic species since the introduction in 1989. We suggest a new aquaculture model of S. intermedius that extends the seed production to November to avoid the mass mortality in summer. In the south, 64% of coastal areas in Zhejiang and Fujian provinces are suitable for the transplantation of S. intermedius to the south. The present study suggests the ecological niche model MaxEnt based on the data of SST and published information as a new tool for the assessment of the site suitability of sea cucumbers and sea urchins in China. This provides new insights into the aquaculture expansion of native and exotic species.

Isolation and characterization of four pathogenic strains of Vibrio spp. from farmed sea urchins, Strongylocentrotus intermedius, in China.

The sea urchin Strongylocentrotus intermedius, famous for its gonadal quality, is one of the most important farmed species in the sea area of northern China. Since 2020, outbreaks of black peristomial membrane disease (commonly called black mouth disease) have frequently occurred in spring and winter in cultured S. intermedius. In this study, we isolated the predominant bacteria from different tissues of diseased sea urchins from a North China farm in the spring of 2021. Four pathogenic strains (named SIBMPM01, SIBMPM02, SIBMPM03 and SIBMCF01) were obtained and characterized by Gram staining, morphological observation, artificial infection tests, and metabolic characteristics. Our results showed that: 1) all obtained strains belonged to the genus Vibrio and had morphological differences. Phylogenetic analysis indicated that the four obtained strains might be novel Vibrio species. 2) Laboratory-based artificial infection tests showed that sea urchins infected with either SIBMPM01, SIBMPM02, SIBMPM03 or SIBMCF01 exhibited pathological symptoms of a black peristomial membrane in a dosage-dependent and temperature-dependent manner. The virulence of SIBMCF01 was greater than those of the others. 3) Metabolic characterization data showed that SIBMPM01, SIBMPM02, SIBMPM03 and SIBMCF01 shared similar metabolic characteristics. 4) Antimicrobial susceptibility tests demonstrated that the four obtained strains were all sensitive to ampicillin, doxycycline, norfloxacin, ofloxacin, furazolidone and chloramphenicol. SIBMPM01 was specifically sensitive to neomycin, and SIBMCF01 was specifically sensitive to carboxybenzyl penicillin.

Exposure of adult sea urchin Strongylocentrotus intermedius to stranded heavy fuel oil causes developmental toxicity on larval offspring.

Heavy fuel oil (HFO) spills pose serious threat to coastlines and sensitive resources. Stranded HFO that occurs along the coastline could cause long-term and massive damage to the marine environment and indirectly affect the survival of parental marine invertebrates. However, our understanding of the complex associations within invertebrates is primarily limited, particularly in terms of the toxicity effects on the offspring when parents are exposed to stranded HFO. Here, we investigated the persistent effects on the early development stage of the offspring following stranded HFO exposure on the sea urchin Strongylocentrotus intermedius. After 21 d exposure, sea urchins exhibited a significant decrease in the reproductive capacity; while the reactive oxygen species level, 3-nitrotyrosine protein level, protein carbonyl level, and heat shock proteins 70 expression in the gonadal tissues and gametes significantly increased as compared to the controls, indicating that HFO exposure could cause development toxicity on offspring in most traits of larval size. These results suggested that the stranded HFO exposure could increase oxidative stress of gonadal tissues, impair reproductive functions in parental sea urchins, and subsequently impact on development of their offspring. This study provides valuable information regarding the persistent toxicity effects on the offspring following stranded HFO exposure on sea urchins.

Direct and latent effects of ocean acidification on the transition of a sea urchin from planktonic larva to benthic juvenile.

Ongoing ocean acidification is expected to affect marine organisms and ecosystems. While sea urchins can tolerate a wide range of pH, this comes at a high energetic cost, and early life stages are particularly vulnerable. Information on how ocean acidification affects transitions between life-history stages is scarce. We evaluated the direct and indirect effects of pH (pHT 8.0, 7.6 and 7.2) on the development and transition between life-history stages of the sea urchin Strongylocentrotus droebachiensis, from fertilization to early juvenile. Continuous exposure to low pH negatively affected larval mortality and growth. At pH 7.2, formation of the rudiment (the primordial juvenile) was delayed by two days. Larvae raised at pH 8.0 and transferred to 7.2 after competency had mortality rates five to six times lower than those kept at 8.0, indicating that pH also has a direct effect on older, competent larvae. Latent effects were visible on the larvae raised at pH 7.6: they were more successful in settling (45% at day 40 post-fertilization) and metamorphosing (30%) than larvae raised at 8.0 (17 and 1% respectively). These direct and indirect effects of ocean acidification on settlement and metamorphosis have important implications for population survival.

Responses of sea urchins (Strongylocentrotus intermedius) with different sexes to CO2-induced seawater acidification: Histology, physiology, and metabolomics.

Responses of different sexes of farmed Strongylocentrotus intermedius to chronic CO2-induced seawater acidification were investigated in 120-day lab-based experiments. Four experimental groups were set up as one control group and three seawater acidification groups. The results showed that 1) Specific growth rate and the numbers of mature gamete cells declined in a pH-dependent way in both sexes of adult S. intermedius. 2) There were differences in SDMs identified in females and males reared in acidified seawater reflecting sex-specific response variation in adult S. intermedius. 3) The number of altered metabolic pathways exhibited a linear increasing trend as seawater pH declined in both sexes of adult S. intermedius. Meanwhile seawater acidification might affect metabolic processes via changing the relative expression and activity of key enzymes controlling the corresponding metabolic pathways of adult S. intermedius.

Macroalgae and interspecific alarm cues regulate behavioral interactions between sea urchins and sea cucumbers.

Sea urchins and sea cucumbers are mutually beneficial organisms in kelp ecosystem. As herbivores, sea urchins process kelp through feeding and egestion, providing inaccessible food for benthic consumers such as sea cucumbers. Sea urchins in turn profit from the sediment cleaned by sea cucumbers. However, behavioral interactions between them remain poorly understood, which greatly hampers our understanding on the relationship between ecologically important benthic species in marine ecosystems and the regulating mechanism. The present study investigated behavioral interactions between sea urchins Strongylocentrotus intermedius and sea cucumbers Apostichopus japonicus in laboratory conditions. We revealed that the presence of sea urchins caused significant higher speed movement of A. japonicus. Interestingly, the negative effects of S. intermedius on A. japonicus were significantly reduced in the shared macroalgal area. For the first time, we found the interspecific responses to alarm cues between sea cucumbers and sea urchins. Conspecific responses were significantly larger than the interspecific responses in both sea urchins and sea cucumbers. This indicates that interspecific response to alarm cues is an efficient approach to anti-predation and coexistence in mutually beneficial organisms. The present study shed light on the interspecific relationships and coexistence between sea urchins and sea cucumbers in kelp ecosystem.

Evidence for association of Vibrio echinoideorum with tissue necrosis on test of the green sea urchin Strongylocentrotus droebachiensis.

"Sea urchin lesion syndrome" is known as sea urchin disease with the progressive development of necrotic epidermal tissue and loss of external organs, including appendages on the outer body surface. Recently, a novel strain, Vibrio echinoideorum has been isolated from the lesion of green sea urchin (Strongylocentrotus droebachiensis), an economically important mariculture species in Norway. V. echinoideorum has not been reported elsewhere in association with green sea urchin lesion syndrome. Therefore, in this study, an immersion based bacterial challenge experiment was performed to expose sea urchins (wounded and non-wounded) to V. echinoideorum, thereby mimicking a nearly natural host-pathogen interaction under controlled conditions. This infection experiment demonstrated that only the injured sea urchins developed the lesion to a significant degree when exposed to V. echinoideorum. Pure cultures of the employed bacterial strain were recovered from the infected animals and its identity was confirmed by the MALDI-TOF MS spectra profiling. Additionally, the hemolytic phenotype of V. echinoideorum substantiated its virulence potential towards the host, and this was also supported by the cytolytic effect on red spherule cells of sea urchin. Furthermore, the genome sequence of V. echinoideorum was assumed to encode potential virulence genes and were subjected to in silico comparison with the established virulence factors of Vibrio vulnificus and Vibrio tasmaniensis. This comparative virulence profile provided novel insights about virulence genes and their putative functions related to chemotaxis, adherence, invasion, evasion of the host immune system, and damage of host tissue and cells. Thus, it supports the pathogenicity of V. echinoideorum. In conclusion, the interaction of V. echinoideorum with injured sea urchin facilitates the development of lesion syndrome and therefore, revealing its potentiality as an opportunistic pathogen.

Interactive effects of ocean acidification, ocean warming, and diurnal temperature cycling on antioxidant responses and energy budgets in two sea urchins Strongylocentrotus intermedius and Tripneustes gratilla from different latitudes.

To accurately predict the fitness of marine ectotherms under the climate change scenarios, interactive effects from multiple environmental stressors should be considered, such as ocean acidification (OA), ocean warming (OW) and diurnal temperature cycling (DTC). In this work, we evaluated and compared the antioxidant capacity and metabolism homeostasis of two sea urchins, viz. the temperate species Strongylocentrotus intermedius and the tropical species Tripneustes gratilla, in response to oceanic conditions under a climate change scenario. The two species were treated separately/jointly by acidic (pH 7.6), thermal (ambient temperature + 3 °C), and temperature fluctuating (5 °C fluctuations daily) seawater for 28 days. The activities of antioxidant enzymes (catalase and superoxide dismutase) and the cellular energy allocation in the urchins' gonads were assessed subsequently. Results showed that exposure to OA, OW, and DTC all induced antioxidant responses associated with metabolism imbalance in both S. intermedius and T. gratilla. The physiological adjustments and energy strategies towards exposure of OA, OW, and DTC are species specific, perhaps owing to the different thermal acclimation of species from two latitudes. Moreover, decrease of cellular energy allocation were detected in both species under combined OA, OW, and DTC conditions, indicating unsustainable bioenergetic states. The decrease of cellular energy allocation is weaker in T. gratilla than in S. intermedius, implying higher acclimation capacity to maintain the energy homeostasis in tropical urchins. These results suggest that climate change might affect the population replenishment of the two sea urchins species, especially for the temperate species.

Gene expression patterns of sea urchins (Strongylocentrotus intermedius) exposed to different combinations of temperature and hypoxia.

Strongylocentrotus intermedius is one of the most economically valuable sea urchin species in China, and its growth and survival are severely constrained by ocean warming and the hypoxia that often accompanies high water temperatures. To elucidate the molecular mechanisms of S. intermedius that regulate gene expression in response to multi-causal environmental stresses. We performed a de novo transcriptome analysis of coelomocyte from S. intermedius to heat (25 °C), hypoxia (2 mg/L), and the combined stress. We identified 35,635, 29,107, and 29,440 differentially expressed genes (DEGs) in S. intermedius cultured under high temperature, low oxygen, and combined stress, respectively. Further Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyses revealed that these DEGs mainly enriched the functional categories of "Protein processing in endoplasmic reticulum," and "Glutathione metabolism" by heat stress, such as HSP70, GSTO1, PDIA4. After hypoxic stress, "Notch signaling pathway" and metabolism-related pathways such as "Glycerolipid metabolism", "Pyruvate metabolism" were significantly enriched. Exposure to combined stress resulted in a two-factor additive effect at the transcriptome level and have a more extensive impact on the immune correlated pathways in S. intermedius than single stress, the expression of related immune genes (C3, C5, and AIFM2) were up-regulated. Quantitative real-time PCR (qRT-PCR) analysis of the expression of 18 DEGs confirmed the RNA-Seq results. Observations in the present study will improve the understanding of the molecular mechanism of S. intermedius in response to multi-causal environmental stress.