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.

miRn-3 inhibits cutaneous wound healing by targeting gelsolin in the sea cucumber Apostichopus japonicus.

The microRNA novel-3 (miRn-3) is a 23-nt small endogenous noncoding RNA of unknown function. To enrich our knowledge of the regulatory function of miRn-3 in the process of wound healing, the sea cucumber Apostichopus japonicus was used as a target model in this study. Gelsolin (AjGSN), a potential target gene of miRn-3, was cloned and characterized, and the interaction between miRn-3 and AjGSN was verified. The function of the miRn-3/AjGSN axis in regulating cutaneous wound healing was explored in the sea cucumber A. japonicus. The results showed that 1) the full-length cDNA of AjGSN was 2935 bp, with a high level of sequence conservation across the echinoderms; 2) miRn-3 could bind to the 3'UTR of AjGSN and negatively regulate the expression of AjGSN; 3) overexpression of miRn-3 and inhibition of the expression of AjGSN suppressed cutaneous wound healing in A. japonicus. In general, all observations of this study suggest that miRn-3 plays an important role in the early process of cutaneous wound healing by negatively targeting AjGSN, and that it may be a potential biomarker in wound healing.

UHPLC-MS/MS metabolomics analysis of sea cucumber (Apostichopus japonicus) processed using different methods.

The effects of different processing methods on the nutritional components of sea cucumber (Apostichopus japonicus) are of concern to consumers who select sea cucumber products. This study employed liquid chromatography tandem mass spectrometry to examine the metabolites in fresh, unsoaked salted, soaked salted, and instant sea cucumber body wall samples sourced from Dalian, China. Metabolites were evaluated utilizing partial least squares discriminant analysis (PLS-DA) and subsequently subjected to KEGG metabolic pathway analysis for further investigation. PLS-DA effectively discriminated the body wall metabolites of sea cucumbers obtained via various processing techniques. The differential metabolites identified predominantly encompassed amino acids, lipids, and carbohydrates. Subsequent KEGG metabolic pathway analysis demonstrated a significant association between lipid, carbohydrate, and amino acid metabolism and the specific processing methods employed. The assessment of nutritional differences corresponding to the various A. japonicus processing methods was conducted. The findings of this study can assist in the choice of sea cucumber products and the selection of suitable processing methods.

An Investigation on the Effects of Dietary Vitamin E on Juvenile Sea Urchin (Strongylocentrotus intermedius): Growth, Intestinal Microbiota, Immune Response, and Related Gene Expression.

A 90 d feeding experiment was conducted to investigate the effects of vitamin E (VE) on growth, intestinal microbiota, immune response, and related gene expression of juvenile sea urchin (Strongylocentrotus intermedius). Six dry feeds were made to contain graded levels of VE (78, 105, 152, 235, 302, and 390 mg/kg); these were named E78, E105, E152, E235, E302, and E390, respectively. Dry feed E50 and fresh kelp (HD) were used as the control diets. There were six replicates of cages in each dietary group, and each cage held 20 sea urchins with an initial body weight of approximately 1.50 g. Results exhibited that weight gain rate and gonadosomatic index (GSI) of the sea urchins were not significantly affected by dietary VE ranging from 78 to 390 mg/kg. Sea urchins in the dry feed groups showed poorer growth performance, but significantly higher GSI than those in the fresh kelp groups. The pepsin and lipase activities were not significantly promoted by low or moderate VE, but were inhibited by a high level of VE (302-390 mg/kg), while amylase and cellulase activities were significantly increased by low or moderate VE, with the highest values observed in the E105 and E235 groups, respectively. VE addition at a low dosage (105-152 mg/kg) showed inhibitory effects on immune and antioxidant enzyme activities and expression of inflammation-related genes, but showed no beneficial effects at moderate or high dosage (235-390 mg/kg), while a moderate or relatively higher level of VE (235-302 mg/kg) significantly increased the expression of several immune-related genes. The relative abundance of Proteobacteria, Actinobacteria, Ruegeria, and Maliponia in the intestine of the sea urchins increased with the increase in VE in the dry feeds. On the contrary, the relative abundance of the Firmicutes, Bacteroidetes, Escherichia-Shigella, Bacteroides, and Clostridium sensu stricto 1 gradually decreased as VE content increased. These results indicated that a moderate level of VE (172.5-262.4) can achieve ideal digestive enzyme activities and growth performance, but a relatively higher level of VE (235-302 mg/kg) was beneficial for maintaining the immune and antioxidant capacity of juvenile S. intermedius by regulating the expression of inflammation- and immune-related genes and abundance of some bacteria to a healthy state.

Chemical Comparison and Identification of Xanthine Oxidase Inhibitors of Dioscoreae Hypoglaucae Rhizoma and Dioscoreae Spongiosae Rhizoma by Chemometric Analysis and Spectrum-Effect Relationship.

Dioscoreae hypoglaucae Rhizoma (DH) and Dioscoreae spongiosae Rhizoma (DS) are two similar Chinese herbal medicines derived from the Dioscorea family. DH and DS have been used as medicines in China and other Asian countries for a long time, but study on their phytochemicals and bioactive composition is limited. This present study aimed to compare the chemical compositions of DH and DS, and explore the anti-xanthine oxidase components based on chemometric analysis and spectrum-effect relationship. Firstly, an HPLC method was used to establish the chemical fingerprints of DH and DS samples, and nine common peaks were selected. Then, hierarchical clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis were employed to compare and discriminate DH and DS samples based on the fingerprints data, and four steroidal saponins compounds (protodioscin, protogracillin, dioscin, gracillin) could be chemical markers responsible for the differences between DH and DS. Meanwhile, the anti-xanthine oxidase activities of these two herbal medicines were evaluated by xanthine oxidase inhibitory assay in vitro. Pearson correlation analysis and partial least squares regression analysis were subsequently used to investigate the spectrum-effect relationship between chemical fingerprints and xanthine oxidase inhibitory activities. The results showed that four steroidal saponins, including protodioscin, protogracillin, methyl protodioscin and pseudoprogracillin could be potential anti-xanthine oxidase compounds in DH and DS. Furthermore, the xanthine oxidase inhibitory activities of the four selected inhibitors were validated by anti-xanthine oxidase inhibitory assessment and molecular docking experiments. The present work provided evidence for understanding of the chemical differences and the discovery of the anti-xanthine oxidase constituent of DH and DS, which could be useful for quality evaluation and bioactive components screening of these two herbal medicines.

A signal recognition particle receptor gene from the sea cucumber, Apostichopus japonicas.

The signal recognition particle (SRP) system delivers approximately 30% of the proteome to the endoplasmic reticulum (ER) membrane. SRP receptor alpha (SRα) binds to SRP for targeting nascent secreted proteins to the ER membrane in eukaryotic cells. In this study, the SRα homologous gene was identified in the sea cucumber, Apostichopus japonicus (AjSRα). AjSRα codes for 641 amino acids and has 54.94% identity with its mammalian homologs. Like mammalian SRα, it is expected to contain the SRP-alpha N domain, SRP54_N domain, and SRP54 domain. In addition, AjSRα is ubiquitously expressed in adult tissues and exhibits a sexually dimorphic expression pattern, with significantly higher expression in ovaries compared to testes. As a maternal factor, AjSRα can be continuously detected during embryonic development. Importantly, we first attempted to investigate its function by using lentiviral vectors for delivering SRα gene-specific shRNA, and we revealed that lentiviral vectors do not induce an upregulation of immune-related enzymes in sea cucumbers. However, compared to the dsRNA-based RNA interference (RNAi) method, lentivirus-mediated RNAi caused dynamic changes in gene expression at a later time. This study supplied the technical support for studying the functional mechanism of SRα in sea cucumbers.

Chromosome-level genome assembly of sea scallop Placopecten magellanicus provides insights into the genetic characteristics and adaptive evolution of large scallops.

Placopecten magellanicus (Gmelin, 1791), a deep-sea Atlantic scallop, holds significant commercial value as a benthic marine bivalve along the northwest Atlantic coast. Recognizing its economic importance, the need to reconstruct its genome assembly becomes apparent, fostering insights into natural resources and generic breeding potential. This study reports a high-quality chromosome-level genome of P. magellanicus, achieved through the integration of Illumina short read sequencing, PacBio HiFi sequencing, and Hi-C sequencing techniques. The resulting assembly spans 1778 Mb with a scaffold N50 of 86.71 Mb. An intriguing observation arises - the genome size of P. magellanicus surpasses that of its Pectinidae family peers by 1.80 to 2.46 times. Within this genome, 28,111 protein-coding genes were identified. Comparative genomic analysis involving five scallop species unveils the critical determinant of this expanded genome: the proliferation of repetitive sequences recently inserted, contributing to its enlarged size. The landscape of whole genome collinearity sheds light on the relationships among scallop species, enhancing our broader understanding of their genomic framework. This genome provides genomic resources for future molecular biology research on scallops and serves as a guide for the exploration of longevity-related genes in scallops.

Effects of Supplementary Kelp Feeding on the Growth, Gonad Yield, and Nutritional and Organoleptic Quality of Subadult Sea Urchin (Strongylocentrotus intermedius) with Soya Lecithin Intake History.

A 23-week feeding experiment was conducted to investigate the effects of supplementary kelp feeding on the growth, gonad development, and nutritional and sensory properties of sea urchin (Strongylocentrotus intermedius) with soya lecithin (SL) intake history. The feeding experiment was divided into experimental phase I and phase II. During phase I, 48 subadult sea urchins (initial weight: 6.28 ± 0.07 g) were fed one of the feeds with different levels of SL (0%, 1.6%, 3.2%) or kelp (Saccharina japonica) for 12 weeks. Then, all sea urchins were fed kelp for the next 11 weeks during the phase II. Each diet was randomly allocated to six cages of sea urchins. The results of phase I showed that weight gain rate (WGR), gonadosomatic index (GSI), gonad sensory properties (color and texture), and essential amino acid (EAA) contents were not significantly affected by SL level in the feed groups. High level (3.2%) of SL suppressed gonad development of S. intermedius with retarded gametogenesis in the 3.2% SL group (stage Ⅱ) compared to those fed 0% and 1.6% SL groups (stage Ⅲ). Sea urchins fed dry feeds exhibited significantly lower WGR and values of color (redness and yellowness) and texture (hardness and gumminess) but higher contents of EAA in the gonads than those fed kelp. The n-3/n-6 polyunsaturated fatty acid (PUFA) and eicosapentaenoic acid (EPA) of gonads in the groups fed with dry feeds showed no significant differences, but were significantly lower than that of kelp group. At the end of phase II, the gonad yellowness and EPA content of gonads in all dry feed groups were significantly increased by supplementary kelp feeding, with a higher increase observed in S. intermedius with SL intake history, while arachidonic acid (ARA) content was significantly improved by supplementary kelp feeding in S. intermedius with SL intake history. Gonad texture was improved to some extent by supplementary kelp feeding. These results indicated that S. intermedius fed dry feeds showed significantly higher GSI and EAA but poorer organoleptic quality and lower n-3/n-6 PUFA and EPA than those fed kelp. Kelp supplementary feeding improved the fatty acid value and organoleptic quality of gonads, especially for the sea urchins with SL intake history.

Swing Regulates Movement Direction in the Sea Cucumber Apostichopus japonicus in the Presence of Food Cue: New Insights into Movement Patterns.

Regulating movement direction is essential in the locomotion of animals. Sea cucumbers, as eyeless animals, do not have eyes for the perception of the surrounding environment and food. They have a unique way of swinging their bodies when a food cue is detected, although they lack an important perceptual tool. The present study investigated the movement patterns of the sea cucumber Apostichopus japonicus in the absence of a food cue (experiment 1) and in the presence of a food cue (experiment 2). In experiment 1, we found that the movement of sea cucumbers was close to a linear motion (motion linearity 0.91 ± 0.01). In experiment 2, sea cucumbers most frequently adjusted the movement direction when being exposed to food (84 times/216 min), indicating that sea cucumbers adjusted the direction of movement in the swing state but not the motion state. In experiment 2, we found significantly lower time in the immobility state in the sea cucumbers in the presence of food cues compared to that of those without being exposed to food cues, and the frequency of the motion state in response to food cues was 1.6 times than that of those without food cue. This suggests that food cues cause the change in motion state in sea cucumbers. Swing frequency was 1.7 times higher in sea cucumbers exposed to food cues than that of those not exposed to food cues. Further, sea cucumbers in the presence of food showed significantly better performances in swing angle and swing velocity compared to those not exposed to food cues. This suggests that food cue significantly affects the swing state of sea cucumbers. Notably, the present study described the movement patterns of sea cucumbers when they detected food cues, and other factors (such as the detection of predators) need to be further studied. The present study provides new insights into the regulation of movement direction in eyeless organisms.

Transcriptome analysis reveals core lncRNA-mRNA networks regulating melanization and biomineralization in Patinopecten yessoensis shell-infested by Polydora.

BACKGROUND: Patinopecten yessoensis, a large and old molluscan group, has been one of the most important aquaculture shellfish in Asian countries because of its high economic value. However, the aquaculture of the species has recently been seriously affected by the frequent outbreaks of Polydora disease, causing great economic losses. Long non-coding RNAs (lncRNAs) exhibit exhibit crucial effects on diverse biological processes, but still remain poorly studied in scallops, limiting our understanding of the molecular regulatory mechanism of P. yessoensis in response to Polydora infestation. RESULTS: In this study, a high-throughput transcriptome analysis was conducted in the mantles of healthy and Polydora-infected P. yessoensis by RNA sequencing. A total of 19,133 lncRNAs with 2,203 known and 16,930 novel were identified. The genomic characterizations of lncRNAs showed shorter sequence and open reading frame (ORF) length, fewer number of exons and lower expression levels in comparison with mRNAs. There were separately 2280 and 1636 differentially expressed mRNAs and lncRNAs (DEGs and DELs) detected in diseased individuals. The target genes of DELs were determined by both co-location and co-expression analyses. Functional enrichment analysis revealed that DEGs involved in melanization and biomineralization were significantly upregulated; further, obviously increased melanin granules were observed in epithelial cells of the edge mantle in diseased scallops by histological and TEM study, indicating the crucial role of melanizaiton and biomineralization in P. yessoensis to resist against Polydora infestation. Moreover, many key genes, such as Tyrs, Frizzled, Wnts, calmodulins, Pifs, perlucin, laccase, shell matrix protein, mucins and chitins, were targeted by DELs. Finally, a core lncRNA-mRNA interactive network involved in melanization and biomineralization was constructed and validated by qRT-PCR. CONCLUSIONS: This work provides valuable resources for studies of lncRNAs in scallops, and adds a new insight into the molecular regulatory mechanisms of P. yessoensis defending against Polydora infestation, which will contribute to Polydora disease control and breeding of disease-resistant varieties in molluscs.

Bacillus licheniformis reduces mortality, improves symptoms, and controls residual pathogenicity in juvenile sea cucumbers Apostichopus japonicus infected with skin ulceration syndrome.

Mass mortality caused by skin ulceration syndrome (SUS) is the bottle-neck for the sustainable aquaculture of the sea cucumber Apostichopus japonicus. In the present study, probiotic Bacillus licheniformis (0.25 × 109 CFU/g) was used as the treatment for A. japonicus infected with the SUS that caused by Vibrio harveyi. We found that B. licheniformis significantly reduced the number of infected sea cucumbers 5 days and 7 days after the treatment (group B), compared to those without B. licheniformis treatment (group C) (P < 0.001; P < 0.001). Further, the sea cucumbers fed B. licheniformis had significantly lower mortality at the end of the experiment (<10%) than that of those without the B. licheniformis treatment (>60%) (P < 0.001). These results suggest that the treatment of B. licheniformis is an effective method to reduce the mass mortality resulted from SUS in sea cucumber aquaculture. Further, 3-5 days of treatment significantly improved the adverse symptoms of SUS on the physiology and behavior of sea cucumbers, including the righting behavior, adhesion behavior, food consumption, fecal output and mobility. This indicates B. licheniformis treatment has the advantage in the recovery of sea cucumbers after SUS. Moreover, there was no significant difference observed in the physiology and behavior of sea cucumbers between the SUS infected sea cucumbers after the 7-day treatment of B. licheniformis and the healthy individuals. SUS infected sea cucumbers effectively returned to a stage of normalcy. Further, we found a significantly lower infected rate in sea cucumbers exposed to the culture water of group B (∼5%) than that of those in exposure to the culture water of group C (∼60%). This indicates that the treatment of B. licheniformis efficiently controls the residual pathogenicity of SUS in culture water. The present study demonstrated the effectiveness of B. licheniformis treatment as an environmentally friendly approach to reducing mortality, improving symptoms, and controlling residual pathogenicity in sea cucumber aquaculture.

Molecular mechanisms that regulate the heat stress response in sea urchins (Strongylocentrotus intermedius) by comparative heat tolerance performance and whole-transcriptome RNA sequencing.

In the context of climate change and extreme high temperature, the commercially important sea urchin Strongylocentrotus intermedius suffers high mortality during summer in Northern China. How sea urchins respond to high temperatures is of great concern to academia and industry. How to understand the heat tolerance of sea urchin from the whole transcriptome level. In this study, the heat-resistant S. intermedius bred by our team and its control group were used as the research objects, then we applied whole-transcriptome RNA sequencing to detect differentially expressed mRNAs, microRNAs, long noncoding RNAs that respond to heat stress in the heat-resistant and control S. intermedius. A competitive endogenous RNA (ceRNA) regulatory network was constructed with predicted pairs of differentially expressed mRNAs and noncoding RNAs and revealed the molecular regulatory mechanisms in S. intermedius responding to heat stress. A functional analysis suggested that the ceRNAs were involved in basal metabolism, calcium ion transport, endoplasmic reticulum stress, and apoptosis. This is the whole-transcriptomic analysis of S. intermedius under heat stress to propose ceRNA networks that will provide a basis for studying the potential functions of long noncoding RNAs and miRNAs in the heat stress response in S. intermedius and provide a theoretical basis for the study of the molecular mechanism of sea urchins in response to environmental changes.

Living in a hypoxic world: A review of the impacts of hypoxia on aquaculture.

Hypoxia is a harmful result of anthropogenic climate change. With the expansion of global low-oxygen zones (LOZs), many organisms have faced unprecedented challenges affecting their survival and reproduction. Extensive research has indicated that oxygen limitation has drastic effects on aquatic animals, including on their development, morphology, behavior, reproduction, and physiological metabolism. In this review, the global distribution and formation of LOZs were analyzed, and the impacts of hypoxia on aquatic animals and the molecular responses of aquatic animals to hypoxia were then summarized. The commonalities and specificities of the response to hypoxia in aquatic animals in different LOZs were discussed lastly. In general, this review will deepen the knowledge of the impacts of hypoxia on aquaculture and provide more information and research directions for the development of fishery resource protection strategies.

Differences in body wall metabolites of Apostichopus japonicus of different ages in liquid chromatography-tandem mass spectrometry.

Apostichopus japonicus is a marine invertebrate with high economic value. Analysis of the effects of age on the structure of this species is important for understanding the status of the population and conservation. In this study, metabolite analysis of the body walls of A. japonicus of three different ages (1, 2, and 3 years) was performed using LC-MS/MS. Metabolite analysis was performed for the KEGG metabolic pathway. The partial least squares-discriminant analysis is clearly distinguished metabolites in the three groups of A. japonicus. A total of 59, 222, and 179 different metabolites were detected in the age 3 vs age 2, age 3 vs age 1, and age 2 vs age 1 controls, respectively (P < 0.05). These metabolic differences involved several metabolic pathways, including phenylalanine metabolism, taurine and hypotaurine metabolism, sulfur metabolism et al. Thiamin, riboflavin, lipoic acid, acetyl l-carnitine levels increase with age, trans-10-heptadecenoic acid, stearic acid, pentadecanoic acid, dibutyl sebacate, and 8,15-diHETE levels decrease with age, which these metabolites can be used as potential markers to determine the age of A. japonicus. Our results provide the foundation for determining the age of A. japonicus and are important for the conservation of germplasm resources of A. japonicus.

Effects of circadian rhythm on behavior and physiology of the sea cucumber Apostichopus japonicus.

The present study investigated the behavioral and physiological responses to the circadian rhythm in the sea cucumber Apostichopus japonicus. We found that righting behavior of sea cucumbers was significantly faster at night than that in daytime (P < 0.001). We thus suggest aqua-farmers carry out seeding at night in stock enhancement. The number of tentacle swings was significantly higher at night than that in daytime (P = 0.005). Thus, we suggest aqua-farmers provide diets before the peak of sea cucumber feeding at night. There was no significant difference in foraging behavior and defecation behavior during the day and at night. This indicates that not all behaviors have different characteristics in circadian rhythm. In addition, we found that cortisol concentration was significantly higher at night than that in daytime (P = 0.021). This suggests that sea cucumbers are probably more prone to be stressed at night. However, there was no significant difference in 5-HT and melatonin during the day and at night, suggesting that 5-HT and melatonin are probably not affected by circadian rhythm. The present study clarifies the behavioral and physiological responses to circadian rhythm and provides valuable information into sea cucumber aquaculture.

Artificial reefs reduce the adverse effects of mud and transport stress on behaviors of the sea cucumber Apostichopus japonicus.

Poor survival of seeds reduces the production efficiency of the sea cucumber Apostichopus japonicus in pond culture. We investigated the effects of sea mud on the movement-related behaviors of A. japonicus with different body sizes. Mud significantly decreased crawling behavior and wall-reaching behavior in small seeds (~ 1 g of body weight), but not in the large ones (~ 2.5 g of body weight). These behaviors were significantly greater in the large seeds of A. japonicus than those in the small individuals when they were both on the mud. This clearly suggests that mud has negative effects on the movement-related behaviors of small seeds, but not on large individuals. We further assessed the effects of inevitable transport stress on the movement-related behaviors of A. japonicus on mud. Significantly poorer performances in crawling behavior, wall-reaching behavior and struggling behavior were observed in stressed A. japonicus (both sizes) than those in unstressed groups. These new findings indicate that transport stress further increases the adverse effects on the movement-related behaviors of A. japonicus on mud. Moreover, we investigated whether adverse effects can be reduced when individuals are directly seeded onto artificial reefs. Crawling behavior, wall-reaching behavior and struggling behavior in stressed A. japonicus (both sizes) seeded onto artificial reefs were significantly greater than those on mud, whereas artificial reefs did not significantly improve the crawling and struggling behaviors of unstressed small seeds. These results collectively indicate that mud and transport stress show negative impacts on the movement-related behaviors of sea cucumbers. Artificial reefs greatly reduce these adverse effects and probably contribute to improving the production efficiency of sea cucumbers in pond culture.

The feces of sea urchins as food improves survival, growth, and resistance of small sea cucumbers Apostichopus japonicus in summer.

Mass mortality and low growth highly decrease the production efficiency and sustainable aquaculture development of the sea cucumber Apostichopus japonicus in summer. Sea urchin feces was proposed to address the summer problems. A laboratory study was conducted for ~ 5 weeks to investigate survival, food consumption, growth and resistance ability of A. japonicus cultured with the feces of sea urchins fed kelp (KF feces, group KF), the feces of sea urchins fed prepared feed (FF feces, group FF), and the prepared sea cucumber feed (group S) at high temperature (25 °C). The sea cucumbers of group KF had better survival (100%) than those of the group FF (~ 84%), higher CTmax (35.9 °C) than those of the group S (34.5 °C), and the lowest skin ulceration proportion (0%) when  they were exposed to an infectious solution among the three groups. These results suggest that the feces of sea urchins fed kelp is a promising diet for improving the survival and enhancing the resistance in A. japonicus aquaculture in summer. Sea cucumbers fed significantly less FF feces after 24 h of ageing than the fresh FF feces, suggesting this kind of feces became unsuitable for A. japonicus in a short time (within 48 h). However, the 24 h of ageing at 25 °C for the high fiber feces of sea urchins fed kelp had no significant effects on the fecal consumption of sea cucumbers. In the present study, both fecal diets provide better individual growth to sea cucumbers than the prepared feed. Yet, the feces of sea urchins fed kelp provided the highest weight gain rate (WGR) to sea cucumbers. Therefore, the feces of sea urchins fed kelp is a promising food to reduce the mortality, to address the problems of summer, and to achieve higher efficiency in A. japonicus aquaculture in summer.

Identification and functional analysis of Dmrt1 gene and the SoxE gene in the sexual development of sea cucumber, Apostichopus japonicus.

Members of the Doublesex and Mab-3-related transcription factor (Dmrt) gene family handle various vital functions in several biological processes, including sex determination/differentiation and gonad development. Dmrt1 and Sox9 (SoxE in invertebrates) exhibit a very conserved interaction function during testis formation in vertebrates. However, the dynamic expression pattern and functional roles of the Dmrt gene family and SoxE have not yet been identified in any echinoderm species. Herein, five members of the Dmrt gene family (Dmrt1, 2, 3a, 3b and 5) and the ancestor SoxE gene were identified from the genome of Apostichopus japonicus. Expression studies of Dmrt family genes and SoxE in different tissues of adult males and females revealed different expression patterns of each gene. Transcription of Dmrt2, Dmrt3a and Dmrt3b was higher expressed in the tube feet and coelomocytes instead of in gonadal tissues. The expression of Dmrt1 was found to be sustained throughout spermatogenesis. Knocking-down of Dmrt1 by means of RNA interference (RNAi) led to the downregulation of SoxE and upregulation of the ovarian regulator foxl2 in the testes. This indicates that Dmrt1 may be a positive regulator of SoxE and may play a role in the development of the testes in the sea cucumber. The expression level of SoxE was higher in the ovaries than in the testes, and knocking down of SoxE by RNAi reduced SoxE and Dmrt1 expression but conversely increased the expression of foxl2 in the testes. In summary, this study indicates that Dmrt1 and SoxE are indispensable for testicular differentiation, and SoxE might play a functional role during ovary differentiation in the sea cucumber.

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.

Non-coding RNAs targeting NF-κB pathways in aquatic animals: A review.

Nuclear factor-kappa B (NF-κB) pathways have a close relationship with many diseases, especially in terms of the regulation of inflammation and the immune response. Non-coding RNAs (ncRNAs) are a heterogeneous subset of endogenous RNAs that directly affect cellular function in the absence of proteins or peptide products; these include microRNAs (miRNAs), long noncoding RNAs (lncRNAs), circular RNAs (circRNAs), etc. Studies on the roles of ncRNAs in targeting the NF-κB pathways in aquatic animals are scarce. A few research studies have confirmed detailed regulatory mechanisms among ncRNAs and the NF-κB pathways in aquatic animals. This comprehensive review is presented concerning ncRNAs targeting the NF-κB pathway in aquatic animals and provides new insights into NF-κB pathways regulatory mechanisms of aquatic animals. The review discusses new possibilities for developing non-coding-RNA-based antiviral applications in fisheries.