Insights into sequence characteristics and evolutionary history of DGATs in arthropods.

Triacylglycerol (TAG) is crucial in animal energy storage and membrane biogenesis. The conversion of diacylglycerol (DAG) to triacylglycerol (TAG) is catalyzed by diacylglycerol acyltransferase enzymes (DGATs), which are encoded by genes belonging to two distinct gene families. Although arthropods are known to possess DGATs activities and utilize the glycerol-3-phosphate pathway and MAG pathway for TAG biosynthesis, the sequence characterization and evolutionary history of DGATs in arthropods remains unclear. This study aimed to comparatively evaluate genomic analyses of DGATs in 13 arthropod species and 14 outgroup species. We found that arthropods lack SOAT2 genes within the DGAT1 family, while DGAT2, MOGAT3, AWAT1, and AWAT2 were absent from in DGAT2 family. Gene structure and phylogenetic analyses revealed that DGAT1 and DGAT2 genes come from different gene families. The expression patterns of these genes were further analyzed in crustaceans, demonstrating the importance of DGAT1 in TAG biosynthesis. Additionally, we identified the DGAT1 gene in Swimming crab (P. trituberculatus) undergoes a mutually exclusive alternative splicing event in the molt stages. Our newly determined DGAT inventory data provide a more complete scenario and insights into the evolutionary dynamics and functional diversification of DGATs in arthropods.

The comparison of morphology and transcriptome in the inner membrane reveals the potential mechanism of the heritable carapace color of the Chinese mitten crab Eriocheir sinensis.

Carapace color plays an important role in the communication, reproduction, and self-defense of crustaceans, which is also related to their economic value. Chinese mitten crab (Eriocheir sinensis) is an important aquaculture species in China, and there are different strains with heritable carapace colors, i.e. Green, White, and Red. However, there is a lack of research on the formation mechanism of carapace color of this species. This study was conducted to compare the histology and transcriptome in the inner membrane of three carapace color strains of E. sinensis. Histological comparisons revealed that the inner membrane of green and red carapace crabs contained more melanin, appearing in clusters, and had a higher presence of yellow or orange pigments. In contrast, the inner membrane of white carapace crabs had smaller and fewer melanin particles, as well as a lower presence of yellow or orange pigments. Observation under an electron microscope showed that the inner membrane of E. sinensis contained a large number of collagen fibers and various types of cells, including fibroblasts, melanocytes, and other tissue cells, which exhibited different levels of activity. Transcriptome analysis showed that the Green, Red, and White strains of E. sinensis had approximately 80.3 K, 81.6 K and 80.3 K expressed unigenes in their inner membranes, respectively. When comparing Green and Red crabs, there were 2, 850 upregulated genes and 2, 240 downregulated genes. In the comparison between Red and White crabs, there were 2, 853 upregulated genes and 2, 583 downregulated genes. Furthermore, there were 2, 336 upregulated genes and 2, 738 downregulated genes in the inner membranes between White and Green crabs. Among these genes, some members of the solute carriers family, which are involved in carotenoid transportation, showed differential expression among the three carapace color strains. Additionally, significant differences were observed in the expression of genes related to melanin synthesis, including wingless/integrate, tyrosinase, guanine nucleotide-binding protein inhibitory subunit, cell adhesion molecule, adenylyl cyclase, and creb-binding protein. there were no differences in the gene expression levels of the crustacyanin family. In conclusion, this study identified several candidate genes associated with carapace color in the inner membrane of E. sinensis, suggesting a close relationship between the heritable carapace colors and the transport of the carotenoids as well as the synthesis of melanin.

Evolution and expression analysis of carotenoid cleavage oxygenase gene family in Chinese mitten crab Eriocheir sinensis.

Carotenoid cleavage oxygenase (CCO) plays a pivotal role in various biological activities, including antioxidant and immune functions in animals. This paper investigates the evolution and expression of CCO genes based on three chordates and 27 arthropods. Aquatic animals exhibit a higher abundance of CCO genes. Despite this, research on CCO in crustaceans has been notably limited, with a complete absence of any previous studies on the CCO genes for the Chinese mitten crab (Eriocheir sinensis). In this study, six CCO genes were identified in the E. sinensis genome database. Results reveal that the evolution of the CCO gene family in Crustacea is primarily characterized by purifying selection, with a preference for employing similar codons. EsCCO1 and EsCCO3 were mainly expressed in the epidermal layer, and EsCCO4 was mainly expressed in the hindgut. Meanwhile, EsCCO5 and EsCCO6 were mainly expressed in the hepatopancreas and endometrium. A notable detail that different EsCCO genes demonstrate distinct expression patterns within various tissues of E. sinensis. The findings of this study offer fundamental insights that could serve as a basis for further exploration into the functions and regulatory mechanisms of CCO genes in crustacean species.

Metabolome and Transcriptome Association Analysis Reveals the Link Between Pigmentation and Nutrition Utilization in the Juveniles of Sea Cucumber Holothuria leucospilota.

The sea cucumber Holothuria leucospilota is an economically and ecologically important tropical species. Following development into juveniles, H. leucospilota undergoes a color change from white to black, involving a pigmentation process for over a period of several months. In this study, a combination of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and Next-Generation sequencing (NGS) were employed to investigate the changes in metabolomic and transcriptomic profiles during pigmentation in H. leucospilota juveniles. The metabolomic analysis identified a total of 341 metabolites, of which 52 were found to be differentially regulated (P < 0.05 and VIP > 1), with 27 being upregulated in white individuals and 25 in black individuals. Additionally, 632 differentially expressed genes (DEGs) were identified, with 380 genes upregulated in white samples and 252 genes upregulated in black samples. Interestingly, the melanin content and tyrosinase transcript levels did not display significant differences between the two groups. Metabolomic data suggested the involvement of the linoleic acid metabolic pathway in pigmentation. Transcriptomic analysis, coupled with realtime PCR validation, revealed a decrease in the transcript levels of digestive enzymes like α-amylase, maltase-glucoamylase, and trehalase after the juveniles changed to black. Furthermore, the mRNA expressions of major yolk proteins showed a decline, indicating a shift in the accumulation of protein nutrient sources. Overall, our findings suggest that during the pigmentation process in H. leucospilota, no significant changes were observed in the classical melanin pathway, while notable alterations were observed in their nutritional status. This study provides valuable insights into the regulatory mechanisms of pigmentation in marine organisms.

Comparative proteomics elucidates the potential mechanism of heritable carapace color of three strains Chinese mitten crab Eriocheir sinensis.

The carapace coloration is important for the environmental adaptation and reproductive behaviors of crustaceans. We selected red, green and white three carapace color strains of Chinese mitten crab (Eriocheir sinensis) strains. These three carapace colors have stable heritability, but the mechanism for their coloration remains unclear.Through histological observations, we have found significant differences in the composition of pigment cells and pigments within the inner membrane of the three color strains, which may be one of the reasons for the color variation. The levels of various carotenoids in both the shell and inner membrane tissues of red and green strains were significantly higher than those of the white strain, while there was no significant difference between the red and green strains. Proteomics studies have identified 2, 034 and 947 different proteins in the shell and inner membrane, respectively. In the shell, there were 18, 13 and 43 differential proteins between red and white strains, green and white strains and green and red strains, respectively. In the inner membrane, there were 44, 24 and 16 differential proteins between red and white strains, green and white strains and green and red strains, respectively. It is clear that the deposited quantity of carotenoids affects the shell formation of three color strains. Some members of the hemocyanin family showed significant variation among different strains. The study yielded two crustacyanin proteins, which were extracted from both the shell and membrane. Of the two proteins, only Crustacyanin-A1 expression showed a difference between the red and green shells strains. In conclusion, these results indicated that the carapace color formation of E. sinensis may be accomplished through pigment binding proteins (PBPs) and pigment cells, which enhance the understanding of color formation mechanism for crustacean.

Functional transcriptome reveals hepatopancreatic lipid metabolism during the molting cycle of the Chinese mitten crab Eriocheir sinensis.

Crustacean molting is highly related to energy and lipid metabolism. This study was conducted to detect the changes of total lipids (TL), triacylglyceride (TAG), phospholipid (PL) and lipid droplets in hepatopancreas, and then to investigate the gene expression patterns related to hepatopancreatic lipid metabolism during the molting cycle of Chinese mitten crab Eriocheir sinensis. Hepatopancreatic TL and TAG increased significantly from post-molt stage to pre-molt stage, then decreased significantly from pre-molt stage to ecdysis stage, which is consistent to the changes of neutral lipid-rich adipocytes in hepatopancreas. By transcriptomic analysis, 65,325 transcripts were sequenced and assembled, and 28,033 transcripts were annotated. Most genes were related to energy metabolism, and the enriched genes were involved in carbohydrate and lipid metabolism and biosynthesis, especially in de novo synthesis of fatty acids and TAG, and ketone body production. Compared to the inter-molt stages, acetyl-CoA carboxylase, fatty acid synthase and other genes related to the synthesis of fatty acids were upregulated in the pre-molt stage. TAG synthesis related genes, including Glycerol-3-phosphate acyltransferase and 1-acylglycerol-3-phosphate acyltransferases, were upregulated in the post-molt stage compared to the inter-molt stage. The expression of ketone body-related genes had no significant changes during the molting cycle. Compared to the TAG synthetic pathway, ketone body biosynthesis may contribute less/secondarily to fatty acid metabolic processes, which could be involved in the other physiological processes or metabolism. In conclusion, these results showed that TAG is the major lipid deposition during inter- and pre-molt stages, and the most genes are related to the fatty acids and TAG metabolism in the hepatopancreas during the molting cycle of E. sinensis.

Control of phosphorus release from sediment by hydrous zirconium oxide combined with calcite, bentonite and zeolite.

This study investigated the effectiveness and mechanism for the control of internal phosphorus (P) liberation from sediment by hydrous zirconium oxide (HZrO2) combined with calcite, bentonite and zeolite. The results suggested that coexisting calcite, calcium-modified bentonite (CaBT) and calcium-modified zeolite (CaZ) all had the ability to promote the adsorption of phosphate (PO43-) onto HZrO2. The mechanisms of PO43- elimination by HZrO2/calcite mixture involved the adsorption of PO43- on calcite, the precipitation of PO43- with Ca2+, and the inner-sphere complexation of PO43- with HZrO2. The amendment of sediment with HZrO2/calcite, HZrO2/CaBT or HZrO2/CaZ mixture can effectively prevent the sedimentary P release, and the immobilization of mobile P in the sediment and the uptake of dissolved reactive P (DRP) from the interstitial water by the amendment material played a key role in the control of P release from sediment by the combined amendment. Capping sediment with HZrO2/calcite, HZrO2/CaBT or HZrO2/CaZ mixture also can effectively intercept sediment P release, and the formation of P static layer attributed to the uptake of interstitial water DRP and DGT (diffusive gradient in thin-films)-unstable P in the upper sediment by the capping material was a key to the inhibition of sedimentary P migration into the overlying water by the combined capping. The great majority of P immobilized by the HZrO2/calcite, HZrO2/CaBT or HZrO2/CaZ combined covering layer is stable P and it has a low re-releasing risk under dissolved oxygen-deficit and pH 5-9 condition. The stability of P bound by the combined covering layer was larger than that by the single HZrO2 covering layer. The results of this research show that the combined use of HZrO2 and calcite, HZrO2 and CaBT, or HZrO2 and CaZ as a capping material has great potential in the reduction of sediment P loading.

Combined amendment and capping of sediment with ferrihydrite and magnetite to control internal phosphorus release.

This study developed novel active capping systems with recycling convenience using ferrihydrite (Fh) combined with magnetite (Mag), and investigated the effectiveness and mechanism for the restriction of endogenous phosphorus movement from sediment into overlying water (OW) by the combined use of Fh and Mag. The Fh/Mag combined amendment effectively hindered endogenous phosphorus release from sediment to OW in dissolved oxygen (DO)-deficit environment, and the immobilization of diffusion gradient in thin film-labile phosphorus (LPDGT) and mobile phosphorus in the sediment played a key role in the control of endogenous phosphorus liberation by the Fh/Mag combined amendment. Combined capping sediment with Fh and Mag effectively hindered endogenous phosphorus release from sediment to OW in anoxic environment, and the inactivation of LPDGT in the upper sediment played a key part in the control of sediment phosphorus release by the Fh/Mag mixture capping. The stability of phosphorus immobilized by the Fh/Mag combined covering layer was related to its construction way, and the majority (around 90%) of P immobilized to the Fh/Mag mixture covering layer had low risk of release in common pH (5-9) and DO-deficit environments. The Fh/Mag mixture amendment or capping did not increase the risk of sediment iron release, and it also did not produce a large impact on the diversity and richness of bacterial community in the sediment. The combined utilization of Fh and Mag as a composite amendment or capping material to prevent the internal phosphorus from being moved to OW can make full use of their respective advantages. The Fh/Mag mixture capping wrapped by permeable fabric has high potential to reduce the risk of endogenous phosphorus from sediment into OW due to its advantages of high internal phosphorus release suppression efficiency, environmental friendliness, application convenience and sustainability.

Occurrence and distribution characteristics of aged microplastics in the surface water, sediment, and crabs of the aquaculture pond in the Yangtze River Delta of China.

The artificial breeding of freshwater crabs in China has become the main source, accounting for 45.69 % of the total output in 2020. However, microplastics widely exist in ponds due to the addition of meals, and the aging and breakage of plastic tools, and people know little about the occurrence of microplastics in the environment and the tissues of crabs during the cultivation of crabs in ponds. In this study, the abundance and characteristics of microplastics in ponds and crabs were studied finely, and the types of microplastics produced by meals and tools and the aging degree of microplastics in different media were studied in a typical aquaculture experimental base in the Yangtze Estuary of China. After we digested all the samples, there were microplastics in the water, sediment, and inedible part of crabs and crab meals, mainly in fiber shape, with a particle size of 100~300µm, and they have a certain degree of aging. The abundance of microplastics in surface water ranges from 4.4 to 10.8 items/L, and that in sediment ranges from 28.6 to 54.3 items/100 g·dry weight sediments. The average abundance of microplastics in crabs was 23.9 ± 15.9 items/individual. The content of microplastics in crabs' intestinal tissue was the highest, followed by gills and hepatopancreas. At the same time, the microplastics found in crabs were positively correlated with crab body weight and negatively correlated with hepatopancreas index. The results show that in the process of artificial breeding pond feeding, microplastics will be released from the process of meals dissolving in water, and fall off due to wear and tear during the use of tools. Microplastics found in the water, sediments and the tissues of crabs were all aged. Humans have a risk of ingesting microplastics when they eat the tissues of nonedible parts of crabs.

Comparative Analyses of Scylla olivacea Gut Microbiota Composition and Function Suggest the Capacity for Polyunsaturated Fatty Acid Biosynthesis.

Although numerous studies in aquatic organisms have linked lipid metabolism with intestinal bacterial structure, the possibility of the gut microbiota participating in the biosynthesis of beneficial long-chain polyunsaturated fatty acid (LC-PUFA) remains vague. We profiled the gut microbiota of the mud crab Scylla olivacea fed with either a LC-PUFA rich (FO) or a LC-PUFA-poor but C18-PUFA substrate-rich (LOCO) diet. Additionally, a diet with a similar profile as LOCO but with the inclusion of an antibiotic, oxolinic acid (LOCOAB), was also used to further demarcate the possibility of LC-PUFA biosynthesis in gut microbiota. Compared to diet FO treatment, crabs fed diet LOCO contained a higher proportion of Proteobacteria, notably two known taxonomy groups with PUFA biosynthesis capacity, Vibrio and Shewanella. Annotation of metagenomic datasets also revealed enrichment in the KEGG pathway of unsaturated fatty acid biosynthesis and polyketide synthase-like system sequences with this diet. Intriguingly, diet LOCOAB impeded the presence of Vibrio and Shewanella and with it, the function of unsaturated fatty acid biosynthesis. However, there was an increase in the function of short-chain fatty acid production, accompanied by a shift towards the abundance of phyla Bacteroidota and Spirochaetota. Collectively, these results exemplified bacterial communities and their corresponding PUFA biosynthesis pathways in the microbiota of an aquatic crustacean species.

Pacific white shrimp (Litopenaeus vannamei) vitelline membrane outer layer protein 1 (VMO1) is produced in the hepatopancreas and transported into ovarian oocytes during vitellogenesis.

In birds, vitelline membrane outer layer protein 1 (VMO1) is an exogenous protein that can be absorbed by eggs as a barrier to prevent the mixing of yolk and egg white. However, researches on VMO1 are limited in birds but not other non-avian species until now. In this study, we first identified a novel Vmo1 cDNA (Lv-Vmo1) in Pacific white shrimp (Litopenaeus vannamei), the most important cultured shrimp in the world. We further analyzed its gene organization, phylogenetic relationship and protein structure. The Lv-Vmo1 transcript was specifically expressed in the hepatopancreas without sexual dimorphism. During ovarian development in female, the hepatopancreatic Lv-Vmo1 mRNA levels showed a significant increase. By in situ hybridization, Lv-Vmo1 mRNA was present in three cell types of the hepatopancreas but neither oocytes nor follicle cells of the ovary. In contrast, immunofluorescence revealed that Lv-VMO1 protein was distributed in the cytoplasms of both hepatopancreatic cells and ovarian oocytes. Western blot showed that Lv-VMO1 protein was produced in the hepatopancreas and transported to the ovary via hemolymph circulation. Identification of a species-specific egg-entry guide protein is the key to the receptor-mediated ovarian transduction of cargo, a novel gene editing approach in oviparous animals. This study lays the mechanism for exogenous transport into penaeid shrimp eggs by VMO1, as a foundation for achieving exogenous protein-mediated incorporation into oocytes.

Comparative transcriptomics reveals the immune dynamics during the molting cycle of swimming crab Portunus trituberculatus.

Molting is one of the most important biological processes of crustacean species, and a number of molecular mechanisms facilitate this complex procedure. However, the understanding of the immune mechanisms underlying crustacean molting cycle remains very limited. This study performed transcriptome sequencing in hemolymph and hepatopancreas of the swimming crab (Portunus trituberculatus) during the four molting stages: post-molt (AB), inter-molt (C), pre-molt (D), and ecdysis (E). The results showed that there were 78,572 unigenes that were obtained in the hemolymph and hepatopancreas of P. trituberculatus. Further analysis showed that 98 DEGs were involved in immunity response of hemolymph and hepatopancreas, and most of the DEGs participated in the process of signal transduction, pattern recognition proteins/receptors, and antioxidative enzymes system. Specifically, the key genes and pathway involved in signal transduction including the GPCR126, beta-integrin, integrin, three genes in mitogen-activated protein kinase (MAPK) signaling cascade (MAPKKK10, MAPKK4, and p38 MAPK), and four genes in Toll pathway (Toll-like receptor, cactus, pelle-like kinase, and NFIL3). For the pattern recognition proteins/receptors, the lowest expression level of 11 genes was found in the E stage, including C-type lectin receptor, C-type lectin domain family 6 member A and SRB3/C in the hemolymph, and hepatopancreatic lectin 4, C-type lectin, SRB, Down syndrome cell adhesion molecule homolog, Down syndrome cell adhesion molecule isoform, and A2M. Moreover, the expression level of copper/zinc superoxide dismutase isoform 4, glutathione peroxidase, glutathione S-transferase, peroxiredoxin, peroxiredoxin 6, and dual oxidase 2 in stage C or stage D significantly higher than that of stage E or stage AB. These results fill in the gap of the continuous transcriptional changes that are evident during the molting cycle of crab and further provided valuable information for elucidating the molecular mechanisms of immune regulation during the molting cycle of crab.

Changes in Protein Degradation and Non-Volatile Flavor Substances of Swimming Crab (Portunus trituberculatus) during Steaming.

To investigate the effect of steaming time (0, 5, 10, 15, 20, and 25 min) on the protein degradation and non-volatile flavor substances of swimming crab (Portunus trituberculatus), the moisture content, total nitrogen (TN), non-protein nitrogen (NPN), free amino acids (FAAs), flavor nucleotides, electronic tongue analysis, and sensory evaluation were determined. The results showed that the contents of NPN and total FAAs were the highest after crabs steamed for 10 min. Meanwhile, the AMP (adenosine monophosphate) content reached the maximum value (332.83 mg/100 g) and the taste active value (TAV) reached 6.67, which indicated that AMP contributes the most to the taste of steamed crab at 10 min. The electronic tongue distinguished the taste difference well, and the sensory score was the highest at 15 min. Combined with equivalent umami concentration (EUC) and TAV value, swimming crab (weight = 200 ± 20 g) steamed for 10-15 min tasted best.

Comparative Transcriptome Analysis Reveals the Process of Ovarian Development and Nutrition Metabolism in Chinese Mitten Crab, Eriocheir Sinensis.

Ovarian development is a key physiological process that holds great significance in the reproduction of the Chinese mitten crab (Eriocheir sinensis), which is an economically important crab species for aquaculture. However, there is limited knowledge for the regulatory mechanisms of ovarian development. To study the molecular mechanisms of its ovarian development, transcriptome analysis was performed in the ovary and hepatopancreas of E. sinensis during ovarian stages I (oogonium proliferation), II (endogenous vitellogenesis), and III (exogenous vitellogenesis). The results showed that 5,520 and 226 genes were differentially expressed in the ovary and hepatopancreas, respectively. For KEGG enrichment analysis, the differentially expressed genes in the ovary were significantly clustered in phototransduction-fly, phagosome, and ECM-receptor interaction. Significantly enriched pathways in the hepatopancreas included fatty acid biosynthesis, fatty acid metabolism, and riboflavin metabolism. Further analysis showed that 25 genes and several pathways were mainly involved in oogenesis, including the ubiquitin-proteasome pathway, cyclic AMP-protein kinase A signaling pathway, and mitogen-activated protein kinase signaling pathway. Twenty-five candidate genes involved in vitellogenesis and endocrine regulation were identified, such as vitellogenin, vitellogenin receptor, estrogen sulfotransferase, ecdysone receptor, prostaglandin reductase 1, hematopoietic prostaglandin D synthase and juvenile hormone acid O-methyltransferase. Fifty-six genes related to nutritional metabolism were identified, such as fatty acid synthase, long-chain-fatty-acid-CoA ligase 4, 1-acyl-sn-glycerol-3-phosphate acyltransferase 4, fatty acid-binding protein, and glycerol-3-phosphate acyltransferase 1. These results highlight the genes involved in ovarian development and nutrition deposition, which enhance our understanding of the regulatory pathways and physiological processes of crustacean ovarian development.

Comparative study of female Chinese mitten crabs based on their sizes and weights.

The Chinese mitten crab (Eriocheir sinensis) is an important aquaculture species in China. While the price of a large crab will generally be 2-5 times higher than that of smaller crabs, it remains unknown whether nutritional quality is affected by market price. To investigate the effect of size on nutritional composition, adult female crabs were collected and assigned to grades I-IV according to decreasing size. The results showed that meat yield and conditional factors were significantly (P < 0.05) reduced with the decreasing size. The different sizes did not significantly (P > 0.05) affect levels of moisture, crude protein (except for hepatopancreatic crude protein), and total lipid. Grade III crabs had the largest hepatopancreatic crude protein level, which was significantly (P < 0.05) increased compared with grade I crabs. A balanced amino acid composition was found in grade IV crabs, while crabs from grades II and IV had the highest essential amino acids score. Levels of highly unsaturated fatty acids, including C22:6n3, and the ratios of n-3 polyunsaturated fatty acids (PUFA)/n-6 PUFA and C22:6n3 (DHA)/C20:5n3 (EPA) in the hepatopancreas were significantly (P < 0.05) increased in grade III crabs compared with the other grades. In conclusion, among the four grades smaller female crabs (average weight: 93-112 g, grades III-IV) displayed an optimal nutritional quality.

Population genetic analysis reveals secondary contact between Eriocheir sinensis and E. japonica in South Korea.

BACKGROUND: The Chinese mitten crab (Eriocheir sinensis) and the Japanese mitten crab (E. japonica) of the family Varunidae, which are also critical fishery species in their native habitats, including China, Japan, and South Korea, exhibit a rare migration life history for the decapod life cycle. Eriocheir sinensis and E. japonica in South Korea may have originated from speciation or secondary contact in South Korea after speciation of these two species; however, the genetic relationship between these South Korean populations remains unclear, and need to be clarified. OBJECTIVE: This study examined the population genetic properties of E. sinensis and E. japonica in South Korea to reveal the origin of their co-existence. METHODS: Mitochondrial DNA cytochrome c oxidase I sequences of 120 E. sinensis and E. japonica from 6 sampling sites in South Korea were sequenced. Their genetic diversity and haplotype data were compared to previously reported data from the Chinese mainland, Taiwan, Japan, and Russia. RESULTS: The haplotype network, analysis of molecular variance, FST values, Mantel test, and Bayesian skyline plot results of South Korean E. sinensis and E. japonica demonstrated that E. sinensis and E. japonica were isolated from their central populations, Chinese and Japanese populations, by the distance due to the temperature, salinity, ocean current flow, or a geohistorical event. In addition, genetic analysis indicated that the South Korean populations of the two species were marginal populations, exhibiting low genetic diversity, significant negative neutrality values, and radial haplotype networks. The current results also suggest that the Geum River is an overlapping habitat for the natural populations of E. sinensis and E. japonica in South Korea. Further research is required to examine the geohistorical and evolutionary event between E. sinensis and E. japonica in South Korea. CONCLUSIONS: The co-existence of E. sinensis and E. japonica in South Korea may have originated from secondary contact after their speciation.

Metabolomics of the hepatopancreas in Chinese mitten crabs (Eriocheir sinensis).

Chinese mitten crab (Eriocheir sinensis) is an important economic seafood in China, and the hepatopancreatic dim color and bitter taste of crabs significantly affects the consumer's acceptance. The crabs with brown hepatopancreas (BH) generally taste bitter, while the orange hepatopancreas (OH) does not taste bitter. While the metabolites perform their physiological functions, their metabolic relationship may directly affect the color and taste quality of the crab hepatopancreas. This study investigated the metabolic relationship of key metabolites related to the color and bitterness in the OH and BH of E. sinensis. The contents of total bile acids (BAs) (10.62 vs. 3.72 µg/g), bitter free amino acids (FAAs) (5.15 vs. 2.46 mg/g) and lutein (6.88 vs. 2.55 µg/g) in the BH group were significantly higher (P < 0.01) than those in the OH group, while ß-carotene (314.47 vs. 478.52 µg/g) in the BH group was significantly lower (P < 0.05) than that in the OH group. In addition, BAs were positively correlated with lutein, saturated and monounsaturated fatty acids (SFA and MUFA) and bitter FAAs, and negatively correlated with color value, ß-carotene and polyunsaturated fatty acids (PUFA). In conclusion, BAs and bitter FAAs may be the major contributors to the bitterness of BH, and the high content of BAs in the BH group may promote lipid catabolism, inhibit ß-carotene absorption and enhance amino acid metabolism, leading to the hepatopancreas brown and bitter.

Genome-Wide Analysis Indicates a Complete Prostaglandin Pathway from Synthesis to Inactivation in Pacific White Shrimp, Litopenaeus vannamei.

Prostaglandins (PGs) play many essential roles in the development, immunity, metabolism, and reproduction of animals. In vertebrates, arachidonic acid (ARA) is generally converted to prostaglandin G2 (PGG2) and H2 (PGH2) by cyclooxygenase (COX); then, various biologically active PGs are produced through different downstream prostaglandin synthases (PGSs), while PGs are inactivated by 15-hydroxyprostaglandin dehydrogenase (PGDH). However, there is very limited knowledge of the PG biochemical pathways in invertebrates, particularly for crustaceans. In this study, nine genes involved in the prostaglandin pathway, including a COX, seven PGSs (PGES, PGES2, PGDS1/2, PGFS, AKR1C3, and TXA2S), and a PGDH were identified based on the Pacific white shrimp (Litopenaeus vannamei) genome, indicating a more complete PG pathway from synthesis to inactivation in crustaceans than in insects and mollusks. The homologous genes are conserved in amino acid sequences and structural domains, similar to those of related species. The expression patterns of these genes were further analyzed in a variety of tissues and developmental processes by RNA sequencing and quantitative real-time PCR. The mRNA expression of PGES was relatively stable in various tissues, while other genes were specifically expressed in distant tissues. During embryo development to post-larvae, COX, PGDS1, GDS2, and AKR1C3 expressions increased significantly, and increasing trends were also observed on PGES, PGDS2, and AKR1C3 at the post-molting stage. During the ovarian maturation, decreasing trends were found on PGES1, PGDS2, and PGDH in the hepatopancreas, but all gene expressions remained relatively stable in ovaries. In conclusion, this study provides basic knowledge for the synthesis and inactivation pathway of PG in crustaceans, which may contribute to the understanding of their regulatory mechanism in ontogenetic development and reproduction.

Comparative proteomics elucidates the dynamics of ovarian development in the Chinese mitten crab Eriocheir sinensis.

Ovarian development is a complex physiological process for crustacean reproduction that is divided into the oogonium proliferation stage, endogenous vitellogenic stage, exogenous vitellogenic stage, and oocyte maturation stage. Proteomics analysis offers a feasible approach to reveal the proteins involved in the complex physiological processes of any organism. Therefore, this study performed a comparative proteomics analysis of the ovary and hepatopancreas at three key ovarian stages, including stages I (oogonium proliferation), II (endogenous vitellogenesis) and IV (exogenous vitellogenesis), of the Chinese mitten crab Eriocheir sinensis using a label-free quantitative approach. The results showed that a total of 2,224 proteins were identified, and some key proteins related to ovarian development and nutrition metabolism were differentially expressed. The 26 key proteins were mainly involved in the ubiquitin/proteasome pathway (UPP), cyclic AMP-protein kinase A (cAMP-PKA) signaling pathway, and mitogen-activated protein kinase (MAPK) signaling pathway during oogenesis. Fifteen differentially abundant proteins (DAPs) were found to participate in vitellogenesis and oocyte development, such as vitelline membrane outer layer protein 1 homolog, vitellogenin, vitellogenin receptor, heat shock 70 kDa protein cognate 3 and farnesyl pyrophosphate synthase. Forty-seven DAPs related to nutrition metabolism were identified, including the protein digestion, fatty acid metabolism, prostaglandin metabolism, lipid digestion and transportation, i.e. short-chain specific acyl-CoA dehydrogenase, acyl-CoA desaturase, fatty acid-binding protein, long-chain fatty acid CoA ligase 4, and hematopoietic prostaglandin D synthase. These results not only indicate proteins involved in ovarian development and nutrient deposition but also enhance the understanding of the regulatory pathways and physiological processes of crustacean ovarian development.