Eriocheir Sinensis Research Articles

Cardiac Transcriptome and Histology of the Heart of the Male Chinese Mitten Crab (Eriocheir sinensis) under High-Temperature Stress

High temperatures are important environmental stressors affecting the metabolism, growth, immunity, and mortality of Chinese mitten crabs (Eriocheir sinensis). In this study, Chinese mitten crabs were divided into two groups and exposed to temperatures of 35 °C (thermal stress group) or 25 °C (control group) for 24 h, and the transcriptome of the heart was analyzed. There were 4007 differentially expressed genes (DEGs) between the thermal stress and the control groups, including 2660 upregulated and 1347 downregulated genes. Heat shock proteins (HSPs) and transcription factors (TFs) were temperature-sensitive DEGs in Chinese mitten crabs. DEGs mainly focused on protein processing in the endoplasmic reticulum, ribosome biogenesis, glycine, serine, and threonine metabolism, protein export, and insect hormone biosynthesis pathways. A total of 28,916 SSRs and 59 TF families, including 851 TFs, were detected among all unigenes of E. sinensis transcripts. The qRT-PCR results for the HSPs and apoptotic DEGs from the heart exhibited the same trends as those in the E. sinensis transcriptome data. Results of light microscopy analyzing histological sections of the heart indicated that most myocardial fibers were lysed, and the number of nuclei and the connective tissue contents between the myocardial layers were both reduced following 35 °C exposure for 24 h.

KIF2A Upregulates PI3K/AKT Signaling through Polo-like Kinase 1 (PLK1) to Affect the Proliferation and Apoptosis Levels of Eriocheir sinensis Spermatogenic Cells.

E. sinensis is an animal model for studying the reproduction and development of crustaceans. In this study, we knocked down the Es-Kif2a gene by injecting dsRNA into E. sinensis and inhibited Es-Plk1 gene expression by injecting PLK1 inhibitor BI6727 into E. sinensis. Then, the cell proliferation level, apoptosis level, and PI3K/AKT signaling expression level were detected. Our results showed that the proliferation level of spermatogenic cells decreased, while the apoptosis level increased after Es-Kif2a knockdown or Es-Plk1 inhibition. In order to verify whether these changes are caused by regulating the PI3K/AKT pathway, we detected the expression of PI3K and AKT proteins after Es-Kif2a knockdown or Es-Plk1 inhibition. Western Blot showed that in both the Es-Kif2a knockdown group and the Es-Plk1 inhibition group, the expression of PI3K and AKT proteins decreased. In addition, immunofluorescence showed that Es-KIF2A and Es-PLK1 proteins were co-localized during E. sinensis spermatogenesis. To further explore the upstream and downstream relationship between Es-KIF2A and Es-PLK1, we detected the expression level of Es-PLK1 after Es-Kif2a knockdown as well as the expression level of Es-KIF2A after Es-Plk1 inhibition. Western Blot showed that the expression of Es-PLK1 decreased after Es-Kif2a knockdown, while there was no significant change of Es-KIF2A after Es-Plk1 inhibition, indicating that Es-PLK1 may be a downstream factor of Es-KIF2A. Taken together, these results suggest that Es-KIF2A upregulates the PI3K/AKT signaling pathway through Es-PLK1 during the spermatogenesis of E. sinensis, thereby affecting the proliferation and apoptosis levels of spermatogenic cells.

Effects of microcystin-LR on immune function, lipid metabolism and intestinal microbial structure in Eriocheir sinensis

To investigate the potential toxic mechanism of Microcystin-LR (MC-LR) on juvenile Eriocheir sinensis (E. sinensis), a toxicity test was conducted by injecting MC-LR into the 3rd pereiopod. Following injection, we evaluated alterations in immune response biomarkers, antioxidant status, lipid metabolism-related genes expression, and intestinal microbiota composition in the hepatopancreas and intestinal tissues of E. sinensis. The results of immune response showed that after injection of MC-LR, the serum complement C3 level in E. sinensis significantly decreased at 48 hours (P < 0.05), whereas complement C4 level increased considerably from 12 to 96 hours (P < 0.01). The phenoloxidase system (PO) showed an increasing trend from 6 to 96 hours (P < 0.05 or P < 0.01). The levels of TNF-α, IL-6 significantly increased at 6 and 12 hours (P < 0.05 or P < 0.01). Gene expression analysis showed that FASN gene in the hepatopancreatic tissue displayed a higher expression at both 6 and 12 hours post-exposure (P < 0.01), while ALDH and ACAA2 gene exhibited a consistent downward trend from 6 to 48 hours (P < 0.05), Additionally, the mRNA levels of MECR and ACSL demonstrated a pattern of initial downregulation, followed by an upregulation, and subsequent downregulation. The antioxidant status results revealed that the levels GSH and CAT increased first and then decreased, while the activity of SOD decreased first and then increased. Meanwhile, the MDA content showed an increasing trend. 16 S rDNA analysis showed that the intestinal microbial diversity in MC-LR treatment group was significantly higher relative to the control group (P < 0.01). Compared with the control group, the MC-LR group had a significantly higher relative abundance of Proteobacteria, Fusobacteria, and Actinobacteria (P < 0.05) and a significantly lower relative abundance of Firmicutes (P < 0.01). At genus level, the relative abundance of Acinnetobacter, Flavobacterium, Acidovorax, Paracoccus, Cloacibacterium, Gemmobacter, Aeromonas, and Dechloromonas in the intestine of MC-LR-treated group was significantly increased relative to the control group (P < 0.05 or P < 0.01), but the relative abundance of Lachnospiraceae_NK4A136_group was significantly decreased (P < 0.01). The results of the study demonstrated that MC-LR could induced oxidative stress, hepatopancreas tissue damage, and intestinal microbial imbalance in E. sinensis.

Effects of dietary L-tryptophan supplementation on agonistic behavior, feeding behavior, growth performance, and nutritional composition of the Chinese mitten crab (Eriocheir sinensis)

In the pond culture of Chinese mitten crabs (Eriocheir sinensis), high stocking densities usually increase the frequency of agonistic behavior, which can directly impact crab integrity, survival, and growth and result in economic losses. We hoped to reduce agonistic behavior by adding additives to feed. In this study, we first observed the agonistic behavior of E. sinensis in 5 molting periods (from first molting to puberty molting) during the adult grow-out stage. Then, we assessed the effects of L-TRP as a feed additive on the agonistic behavior of juvenile, precocious, subadult, and adult crabs. Finally, we performed 106 days of cage culture to assess the effects of L-TRP on feeding behavior, animal vitality, growth performance, and nutritional composition. The results showed that the intensity of agonistic behavior was significantly higher after puberty molting than in other molting periods during the adult grow-out stage. Moreover, L-TRP supplementation inhibited the agonistic behavior of juvenile, subadult, and adult crabs, but it did not affect the precocious crabs. During cage culture, L-TRP significantly improved crab vitality after 3 weeks but not after 6 and 9 weeks, but it reduced the ingestion rate after 9 weeks. Moreover, the L-TRP group showed a significant increase in the weight gain rate and specific growth rate after the fourth molting. After 106 days of culture, the content of proline and fatty acids in the muscle decreased significantly after L-TRP diet supplementation. Some long-chain polyunsaturated fatty acids also decreased significantly in the hepatopancreas. However, we must focus on the adverse effects of L-TRP on Pro and fatty acid composition. Further research is required on the balance between inhibition of agonistic behavior and effects of L-TRP on the nutritional quality of E. sinensis.

Exposure to nanopolystyrene and phoxim at ambient concentrations causes oxidative stress and inflammation in the intestines of the Chinese mitten crab (Eriocheir sinensis)

Nanopolystyrene (NP) and phoxim (PHO) are common environmental pollutants in aquatic systems. We evaluated the toxic effects of exposure to ambient concentrations of NP and/or PHO in the intestines of the Chinese mitten crab (Eriocheir sinensis). Our study showed that histopathological changes were observed in the intestines. Specifically, NP and/or PHO exposure increased intraepithelial lymphocytes. Furthermore, NP and/or PHO exposure induced oxidative stress, as evidenced by a significant decrease in superoxide dismutase activity (SOD), peroxidase activity (POD), and total antioxidant capacity (T-AOC). Pro-inflammatory gene expression and transcriptome analysis demonstrated that NP and/or PHO exposure induced the intestinal inflammatory response. Transcriptome results showed that NP and/or PHO exposure upregulated the NF-κB signaling pathway, which is considered a key pathway in the inflammatory response. Additionally, the expression of pro-inflammatory genes significantly increased after a single exposure to NP or PHO, but it exhibited a significant decrease after the co-exposure. The downregulation of these genes in the co-exposure group likely suggested that the co-exposure mitigated intestinal inflammation response in E. sinensis. Collectively, our findings mainly showed that NP and/or PHO exposure at ambient concentrations induces oxidative stress and inflammatory response in the intestines of E. sinensis.

Effect of temperature on the occurrence of Metschnikowia bicuspidata disease in Eriocheir sinensis

Eriocheir sinensis aquaculture has been significantly impeded by “milky disease,” which is caused by Metschnikowia bicuspidata. This disease exhibits distinct seasonality and is closely associated with temperature fluctuations. The present study investigated the impact of temperature on the proliferation and pathogenicity of M. bicuspidata and the immune capacity of E. sinensis. We observed a significant temperature-dependent effect on the proliferation rate of M. bicuspidata in vitro and in vivo. Although low temperatures (4 °C) suppressed M. bicuspidata proliferation, different temperatures did not have a considerable influence on pathogenicity (P > 0.05). Cultivation at lower temperatures significantly reduced the proliferation rate of pathogens in infected E. sinensis and the time to death. Further, the bacteriostatic capacity of E. sinensis serum was significantly enhanced under high temperatures compared with that under low-temperature conditions (P < 0.01). Transcriptome analysis of hemocytes from E. sinensis at different breeding temperatures indicated that the host's metabolism and immunity were differentially affected at 4 °C compared to 32 °C, with pathways associated with growth and development primarily affected under low-temperature conditions and pathways related to energy supply predominantly influenced under high-temperature conditions. Notably, temperature also led to changes in the expression of genes closely linked to immunity and apoptosis, such as acid phosphatase, C-type lectin, glutathione peroxidase, caspase, and initiator caspase-8. The current findings have significant implications for understanding the effect of temperature on M. bicuspidata pathogenicity in E. sinensis and preventing milky disease infection during aquaculture.

The interaction between lipid and vitamin D3 impacts lipid metabolism and innate immunity in Chinese mitten crabs Eriocheir sinensis

As a fat-soluble vitamin, vitamin D3 relies on fat to perform its biological function, affecting lipid metabolism and innate immunity. This study used different percentages of lipid and vitamin D3 diets to evaluate the synergistic effects on the growth, lipid metabolism and immunity of juvenile Eriocheir sinensis (5.83 ± 0.01 g) for 56 days, including low lipid (LL, 1.5%) and normal lipid (NL, 7.5%) and three levels of vitamin D3: low (LVD, 0 IU/kg), medium (MVD, 9000 IU/kg) and high (HVD, 27,000, IU/kg). The synergistic effect of lipid and vitamin D3 was not significant on growth but significant on ash content, total protein, hepatopancreas lipid content, hemolymph 1α,25-hydroxy vitamin D3 [1α,25(OH)2D3] content, hepatopancreas lipolysis and synthesis genes. Crabs fed normal lipid (7.5%) and medium vitamin D3 (9000 IU/kg) had the highest hepatopancreas index, hemolymph 1α,25(OH)2D3 content, antibacterial ability, immune-related genes and hepatopancreatic lipid synthesis genes expression, but down-regulated the lipolysis genes expression. In contrast, crabs fed diets with low lipid percentage (1.5%) had low growth performance, hemolymph 1α,25(OH)2D3, mRNA levels of lipid synthesis genes, antibacterial ability and immune-related gene expression. At the 1.5% lipid level, excessive or insufficient vitamin D3 supplementation led to the obstruction of ash and protein deposition, reduced growth and molting, aggravated the reduction in antioxidant capacity, hindered antimicrobial peptide gene expression and reduced innate immunity, and resulted in abnormal lipid accumulation and the risk of oxidative stress. This study suggests that diets' lipid and vitamin D3 percentage can enhance antioxidant capacity, lipid metabolism and innate immunity in E. sinensis. A low lipid diet can cause growth retardation, reduce antioxidant capacity and innate immunity, and enhance lipid metabolism disorder.

The effect of fermented feed (FF) on intestinal, surrounding water, and sediment microbiomes of the Chinese mitten crab Eriocheir sinensis

AbstractObjectiveFermented feed (FF) has several significant advantages in aquaculture. However, little is known about the characteristics of intestinal and environmental microbiota in the Chinese mitten crab Eriocheir sinensis.MethodsThis study was conducted to elucidate the relationship between bacterial communities in the intestine of Chinese mitten crabs and the surrounding environment and to compare the bacterial communities in FF and non‐FF diet modes by using 16S ribosomal RNA high‐throughput sequencing technology.ResultFermented feed diets mainly influenced the alpha diversity of intestinal microbiomes but could affect microbial community structures of beta diversity in water and sediment. Compared to water and sediment, crab intestines had the lowest operational taxonomic unit (OTU) richness and bacterial community diversity. The OTU richness and Chao richness index values were lower in the intestines of crabs that received FF than in crabs that were given non‐FF. No significant differences were observed in the bacterial diversity and OTU richness of water and sediment between the two feeding modes. The bacterial community structures were significantly different among the three environments. Tenericutes, Proteobacteria, and Actinobacteria were the most abundant phyla in the crab intestine, sediment, and water, respectively. Bacterial functions were also different among the three environments. Proportions of nitrate reduction and fermentation in the crab intestine, proportions of nitrification and sulfate respiration in the sediment, and proportions of chemoheterotrophy and aerobic chemoheterotrophy in the water were higher for the FF feeding mode than for the non‐FF mode.ConclusionThe bacterial community structures were significantly different among the three environments. The microbiome of the crab intestine was partially shared with that of the ambient environment. Additionally, the FF mainly influenced the alpha diversity of intestinal microbiomes but also affected the community structures of beta diversity in water and sediment. These results will help us to devise dietary strategies to influence the crab intestinal microbiota and provide guidance for sustainable crab culture practices.

Replacement of fish meal with cottonseed protein concentrate in Chinese mitten crab (Eriocheir sinensis): Nutrient digestibility, growth performance, free amino acid profile, and expression of genes related to nutrient metabolism

This study aimed to investigate the application of cottonseed protein concentrate (CPC) in Chinese mitten crabs (Eriocheir sinensis). First, the apparent digestibility coefficient (ADC) of CPC, fish meal and soybean meal were compared in crabs (21.72 ± 0.33 g). The protein ADC of CPC was 90.42%, which was significantly higher than that of soybean meal (83.16%) (P < 0.05). The ADC of Phe, Cys and Glu of CPC were significantly higher than those of fish meal, while the ADC of Ile, Leu, Lys, Met, Thr and Ala of CPC were significantly lower (P < 0.05). Second, we investigated the effects of fish meal substitution by CPC on growth performance, free amino acid profile, and expression of genes related to nutrient metabolism in crabs. Six diets were formulated by replacing 0%, 15%, 30%, 45%, 60% and 75% fish meal with CPC, namely FM, CPC15, CPC30, CPC45, CPC60, and CPC75. A total of 630 crabs (1.68 ± 0.00 g) were randomly divided into 18 tanks (3 tanks per group) and fed 3 times daily for 9 weeks. Results showed that CPC75 group significantly reduced growth performance, feed conversion efficiency, and free Ile, Leu, Lys, Met, and Thr contents in muscle (P < 0.05). The contents of free amino acids (Arg, His, Ile, Leu, Lys, Met, Phe, Thr, Val, Ala, Cys, Glu, Gly, Ser and Tyr) in hepatopancreas decreased linearly with the increase of dietary CPC level (P < 0.05). The substitution of more than 45% fish meal with CPC significantly decreased the concentration of delicious amino acids (Ala, Glu and Gly) in hepatopancreas (P < 0.05), which might adversely affect crab flavor. The expression of genes related to antioxidant capacity, protein transport, TOR pathway and lipid metabolism was significantly downregulated by increasing dietary CPC level (P < 0.05). In conclusion, based on the quadratic regression analysis of FCR and PER, the optimal replacement levels of fish meal with CPC in crab diet containing 35% fish meal were 32.36% and 35.38%, respectively. It is recommended that Ile, Leu and Thr be supplemented in addition to Met and Lys in the application of CPC.

The P4' Peptide-Carrying Bacillus subtilis in Cottonseed Meal Improves the Chinese Mitten Crab Eriocheir sinensis Innate Immunity, Redox Defense, and Growth Performance.

This study developed a recombinant Bacillus subtilis to carry the LGSPDVIVIR peptide (cmP4) isolated from the hydrolyzed products of cottonseed meal with excellent antioxidant and immune-enhancing properties in vitro. It was carried as a tandem of five cmP4 peptides (cmP4') to be stably expressed on a large scale. Then, its effectiveness was evaluated in Chinese mitten crab (Eriocheir sinensis) based on growth performance, redox defense, and innate immunity. A total of 280 crabs (mean body weight: 41.40 ± 0.14) were randomly assigned to seven diets including a control one (without B. subtilis) and six experimental ones with different doses (107,108, and 109 CFU/kg) of unmodified and recombinant B. subtilis, respectively, for 12 weeks. Each diet was tested in four tanks of crabs (10/tank). In terms of bacterial dosages, the final weight (FW), weight gain (WG), hemolymph and hepatopancreatic activities of superoxide dismutase (SOD), catalase (CAT), lysosome (LZM), acid phosphatase (ACP) and alkaline phosphatase (AKP), and hepatopancreatic transcriptions of cat, mitochondrial manganese superoxide dismutase (mtmnsod), thioredoxin-1 (trx1), and prophenoloxidase (propo) all increased significantly with increasing B. subtilis dosages, while hemolymph and hepatopancreatic malondialdehyde (MDA) content and the transcriptions of toll like receptors (tlrs), NF-κB-like transcription factor (relish), and lipopolysaccharide-induced TNF-α factor (litaf) all decreased remarkably. In terms of bacterial species, the recombinant B. subtilis group obtained significantly high values of FW, WG, hemolymph, and hepatopancreatic activities of SOD, CAT, LZM, ACP, and AKP, and the transcriptions of mtmnsod, peroxiredoxin 6 (prx6), and propo compared with the unmodified B. subtilis, while opposite results were noted in hemolymph and hepatopancreatic MDA content and the transcriptions of tlrs, relish, and litaf. These results indicated that dietary supplementation with 109 CFU/kg of recombinant B. subtilis can improve the growth performance, redox defense, and nonspecific immunity of E. sinensis.

Assessing the ecotoxicity of florfenicol exposure at environmental levels: A case study of histology, apoptosis and microbiota in hepatopancreas of Eriocheir sinensis

The intensification of production practices in the aquaculture industry has led to the indiscriminate use of antibiotics to combat diseases and reduce costs, which has resulted in environmental pollution, posing serious threats to aquaculture sustainability and food safety. However, the toxic effect of florfenicol (FF) exposure on the hepatopancreas of crustaceans remains unclear. Herein, by employing Chinese mitten crab (Eriocheir sinensis) as subjects to investigate the toxic effects on histopathology, oxidative stress, apoptosis and microbiota of hepatopancreas under environment-relevant (0.5 and 5 μg/L), and extreme concentrations (50 μg/L) of FF. Our results revealed that the damage of hepatopancreas tissue structure caused by FF exposure in a dose-and time-dependent manner. Combined with the increased expression of apoptosis-related genes (Caspase 3, Caspase 8, p53, Bax and Bcl-2) at mRNA and protein levels, activation of catalase (CAT) and superoxide dismutase (SOD), and malondialdehyde (MDA) accumulation, FF exposure also induced oxidative stress, and apoptosis in hepatopancreas. Interestingly, 7 days exposure triggered more pronounced toxic effect in crabs than 14 days under environment-relevant FF concentration. Integrated biomarker response version 2 (IBRv2) index indicated that 14 days FF exposure under extreme concentration has serious toxicity effect on crabs. Furthermore, 14 days exposure to FF changed the diversity and composition of hepatopancreas microbiota leading remarkable increase of pathogenic microorganism Spirochaetes following exposure to 50 μg/L of FF. Taken together, our study explained potential mechanism of FF toxicity on hepatopancreas of crustaceans, and provided a reference for the concentration of FF to be used in culture of Chinese mitten crab.

Effects of dietary supplementation with Haematococcus pluvialis on the growth, antioxidantase activities, and gut microbiota of Chinese mitten crab (Eriocheir sinensis)

AbstractThe Chinese mitten crab (Eriocheir sinensis) is bred extensively in Eastern and Northern China and faces a series of problems such as frequent diseases, germplasm degradation, and declining benefits, which have appeared to seriously affect the sustainable development of the cultivation industry. Haematococcus pluvialis, a type of unicellular green algae, have been proven to be the suitable dietary supplementation in crustacean feeds. After 14 days of feeding with 5% dietary supplementation with Haematococcus pluvialis powder, the growth, antioxidantase activities, and gut microbiota of E. sinensis were analyzed. The results indicated that H. pluvialis powder could significantly improve growth indicators including weight gain rate (WGR) and specific growth rate (SGR) of E. sinensis (p &lt; 0.05). The levels of superoxide dismutase (SOD) and catalase (CAT) declined, while that of phenoloxidase (PO) kept rising under the treatment of H. pluvialis powder. Moreover, alkaline phosphatase (AKP) activity dropped initially and then increased. No significant differences in the Simpson index and Chao1 index in the control and H. pluvialis powder treatment were observed (p &gt; 0.05). Although the dominant microbiota in the crab gut were not affected significantly by H. pluvialis powder, significant differences in the relative abundance of the dominant phyla between the control and H. pluvialis powder treatment groups were observed (p &lt; 0.05). Furthermore, the relative abundance of Firmicutes and Candidatus_Bacilloplasma, which play an important role in the life activities of E. sinensis, were significantly higher in the group treated by H. pluvialis powder than in the control (p &lt; 0.05). Therefore, the H. pluvialis powder could be potentially used not only as a rapid fattening regulator, but also to improve the immunity and metabolic capacity of E. sinensis.

Effects of feed protein levels on Chinese mitten crabs (Eriocheir sinensis) under the rice-crab co-culture model: Performance, nutrient composition, antioxidant capacity and immunity

The Chinese mitten crab is an important aquaculture product in China. Currently, rice-crab co-culture is a novel model that can realize positive interactions. The feed for Chinese mitten crabs cultured in rice fields should be distinguished from other modes, since their culture environment is unique. Therefore, this study designed three experimental groups that were fed 15% (RC1), 25% (RC2), and 35% (RC3) protein, with a natural bait group (RC0) without artificial feeding as the control. The protein nutritional requirements of Chinese mitten crabs in a rice-crab co-culture model were investigated by exploring the indicators of growth performance, nutrient composition, immunity, antioxidant capacity, and economic benefits of different groups. Through a comprehensive discussion of one-way analysis of variance and regression analysis of the data, we obtained the following results: feeds with high-level protein improved the growth performance, quality, and economic benefits of Chinese mitten crabs and reduced their exposure to oxidative stress. Specifically, the contents of major nutrients and flavor-presenting substances, such as free amino acids (FAA), nucleotides, and essential amino acids (EAA), in the edible parts of female crabs in the 35% protein group were significantly higher than those in all other groups. Male crabs fed 25% − 28.08% protein diets showed higher production performance and FAA content. In conclusion, the optimal dietary protein levels for male and female crabs reared in rice fields are different: they are 35% for females, and at least 25% for males. Therefore, for actual crab production, male and female crabs could be cultured separately using different protein baits to reduce bait costs and achieve green and sustainable development.

Effect of dietary β-carotene supplementation on growth and antioxidant capability of pre-adult male Chinese mitten crab, Eriocheir sinensis

β-carotene has been shown to be beneficial for the color formulation, survival, antioxidation, immunity and stress resistance of many crustaceans. This study was conducted to investigate the effects of dietary supplementation of synthetic β-carotene on growth, antioxidant status, high pH resistance, color parameters, and carotenoids composition of pre-adult Chinese mitten crab Eriocheir sinensis. Three diets were formulated to contain 3.06, 52.40 and 103.59 mg/kg dry diets of synthetic β-carotene (defined as Diet 1 - 3). The results showed that: (1) There were no marked differences in body weights or feed utilization, suggesting that β-carotene was not particularly efficient in enhancing the growth of pre-adult E. sinensis. (2) Indicators of antioxidant capacity (T-AOC, MDA) and immunity (AKP, ACP) both got improved in the hepatopancreas and hemolymph with the supplementation of 50–100 mg/kg β-carotene in the diets. (3) The stress test with high pH indicated Diet 1 treatment had the highest mortality among all treatments, and the lowest mortality was found in Diet 3 treatment. (4) For color enhancement, the redness of the carapace and hepatopancreas increased significantly with the increase of dietary β-carotene supplementation at Day 30. Combined, the inclusion of β-carotene in the diet could enhance the antioxidative capability, non-specific immunity, stress resistance to high pH and tissue carotenoid contents, and the optimal level of dietary synthetic β-carotene supplementation was suggested to be around 50 mg/kg for pre-adult male E. sinensis.

Dietary N-carbamylglutamate (NCG) supplementation improves the survival, digestive and immunological parameters by promoting ammonia excretion in Chinese mitten crab (Eriocheir sinensis), under ammonia nitrogen stress

Ammonia nitrogen is one of the most common environmental stress factors in aquaculture. The aim of this experiment was to evaluate the effects of ammonia nitrogen stress on ammonia metabolism, digestion and immune function as well as the protective effects of dietary N-carbamylglutamate (NCG) in the Chinese mitten crab (Eriocheir sinensis). Two feeds were used, namely a control feed (regular commercial feed) and an NCG feed (NCG (302.96 ± 4.07) mg/kg added). Subsequently, animals fed by these two feeds were exposed to 30 mg/L or 60 mg/L ammonia nitrogen stress (named A30 or A60) for 21 days, respectively. The rate of hemolymph ammonia excretion was significantly increased in NCG group compared to C (control) group (P < 0.05). Hemolymph ammonia levels were significantly lower in NCG A30 (NCG feed and A30) and NCG A60 (NCG feed and A60) groups compared to A30 (control feed and A30) and A60 (control feed and A60) groups (P < 0.05). Glutamate dehydrogenase and glutamine synthetase activities in the gills and hepatopancreas of NCG A60 group were significantly increased compared with the A60 group, which promoted glutamine synthesis (P < 0.05). The cumulative mortality rate of Eriocheir sinensis in NCG A30 and NCG A60 groups were effectively reduced (P < 0.05). The arginine levels of hepatopancreas were significantly increased in NCG A60 group (P < 0.05). Under ammonia nitrogen stress, NCG significantly increased the activities of trypsin, pepsin, lipase and α-Amylase in NCG feed group, and promoted feeding (P < 0.05). In hemolymph, hepatopancreas and intestine, NCG significantly increased nitric oxide synthase activity, lysozyme and phenol oxidase activities compared to the ammonia nitrogen stress group (P < 0.05). The results indicated that, NCG added in feed could decelerate the toxic effects of ammonia on Eriocheir sinensis by promoting ammonia excretion and accelerating glutamine and endogenous arginine synthesis by metabolism. Moreover, the activity of digestive enzymes and immune-related enzymes is improved.

Different types of dietary carotenoids improve the color and odor quality of Eriocheir sinensis ovaries

Chinese mitten crab, Eriocheir sinensis, is a commercially ecological crab species in China. Ovaries are one of the major edible parts for female E. sinensis, and their color and flavor primarily affect consumer acceptance. This study investigated the effects of different dietary carotenoids on the color and odor quality of adult female E. sinensis ovaries. The crab was fed the experiment diets with supplementation of 100 ×10−6 of β-carotene, astaxanthin, lutein, canthaxanthin and zeaxanthin, respectively, for 70 days, while the control diet without the supplementation of carotenoids. The results showed that different carotenoids generated an orange-red coloration on crab ovaries, and astaxanthin and canthaxanthin significantly increased the redness (P < 0.05). All carotenoid groups had higher total carotenoids and fatty acid contents than the control group. Moreover, β-carotene significantly promoted ovarian carotenoid deposition, and lutein predominantly affected fatty acid accumulation (P < 0.05). β-carotene and lutein groups accumulated more aroma components (alcohols, ketones, and aldehydes) by regulating the metabolism of carotenoid and fatty acids in ovaries. These results show that canthaxanthin is a potential astaxanthin substitute for promoting coloration, while β-carotene and lutein can improve the odor quality of the ovaries of adult female E. sinensis.

The Role of Cholesterol during the Ovarian Maturation and Lipid Metabolism of Female Chinese Mitten Crab (Eriocheir sinensis)

In previous study, we found that the cholesterol requirement of Eriocheir sinensis was 0.27%, to further investigate the effects of cholesterol on health status, ovarian maturation, and lipid metabolism of female Eriocheir sinensis broodstock. Two diets containing 0% and 0.25% (actually 0.05% and 0.27%) cholesterol were fed to the female crabs (average weight: 49.21 ± 0.11 g) for 4 months and sampled once a month. The results showed that the body weight (BW), survival rate (SR), meat yield (MY), condition factor (CF), hepatosomatic index (HSI), and gonadosomatic index (GSI) of Eriocheir sinensis were significantly affected by treatment time and compared with the cholesterol deficient group, supplementing cholesterol significantly increased BW, HSI, and GSI (P&lt;0.05). In addition, long-term lack of cholesterol will lead to a significant decrease in the activity of ACP, AKP, and SOD and a significant increase in the content of MDA. The histological results showed that cholesterol significantly increased the volume of oocytes (P&lt;0.05). Further studies found that 0.27% cholesterol significantly increased the transcription levels of vtg and vgr in hepatopancreas and ovaries, which may be the main reason for the increase of oocyte size (P&lt;0.05). When fed with 0.27% cholesterol diet, the contents of nutrients in hepatopancreas and ovaries increased significantly, especially lipids and cholesterol (P&lt;0.05). Through the analysis of mRNA expression level of genes related to lipid metabolism, it was found that cholesterol enhanced the transcription level of genes related to lipid synthesis and transport in hepatopancreas, thereby promoting the accumulation of lipid in the organism. Furthermore, compared with control group, the levels of juvenile hormone (JH), 17β-estradiol (E2), methyl farnesoate (MF), and ecdysone in the organism were significantly increased after feeding a diet with 0.27% cholesterol (P&lt;0.05). In summary, supplementing an appropriate amount of cholesterol in the diet can improve the growth performance of Eriocheir sinensis broodstock, enhance the body’s antioxidant and immune system, and promote the accumulation of nutrients in the ovaries, thereby promoting ovarian maturation.

Effects of Antarctic Krill Meal in Diet on Reproductive Performance and Embryo Quality of Eriocheir sinensis.

A 60-day feeding trial was conducted to evaluate the impact of dietary Antarctic krill meal on the reproductive performance and embryo quality of the Chinese mitten crab, Eriocheir sinensis. Three diets were formulated, incorporating varying levels of Antarctic krill meal at 0% (Diet K0), 10% (Diet K10), and 20% (Diet K20), with a control group fed razor clam Sinonovacula constricta. Each diet was randomly assigned to three replicate tanks, each stocked with 5 males and 10 females. Male and female weights were 145.38 ± 8.01 and 102.57 ± 9.73 g, respectively. The results revealed no significant differences in weight gain rate, specific growth rate, and survival rate. However, the hepatopancreatic weight and hepatopancreas index of female crabs in each group decreased, while gonadal weight and gonadosomatic index increased significantly after 60 days, with Diet K20 showing the highest values. Egg production and fecundity of female crabs reached their peak in Diet K20, with no significant differences in reproductive indices among all groups. The phospholipid content in Diet K20 was significantly higher than in the other groups (P < 0.05). Cholesterol contents in Diet K0 and the control group were significantly higher than in Diet K10 and K20 (P < 0.05). No significant differences were observed in egg diameter, egg weight, moisture, crude protein, and crude fat between the groups. The content of C20 : 2 and C20 : 4n6 was highest in Diet K0, with a significant difference compared to Diet K10 (P < 0.05). However, no significant differences were found in the total content of saturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids among all groups. Based on the research findings, it is recommended that the optimal level of Antarctic krill meal in diets is 20%.

Impact Analysis of Photoperiodic Disorder on the Eyestalk of Chinese Mitten Crab (Eriocheir sinensis) through High-Throughput Sequencing Technology.

Light is an indispensable factor in the healthy growth of living organisms, and alterations in the photoperiod can have consequences for body homeostasis. The eyestalk is a photosensitive organ that secretes various hormones to regulate the Chinese mitten crab (Eriocheir sinensis). However, the photoperiod-dependent eyestalk patterns of gene expression that may underlie changes in body homeostasis are unknown. In this study, we investigated the molecular mechanisms involved in eyestalk transcriptomic responses in E. sinensis under different photoperiod regimes on days 2, 4, and 6. The photoperiods tested were 12, 24, and 0 h light/day. In total, we obtained 110, 958, 348 clean datasets and detected 1809 differentially expressed genes (DEGs). Genes involved in the crustacean hyperglycemic hormone superfamily and juvenile hormones were observed, which play important roles in gonadal development, growth, and immunity in E. sinensis and may also be involved in photoperiod adaptation. In addition, the MAPK signaling pathway was the only signaling pathway identified in the continuous light group but was absent in the continuous darkness group. We suggest that the MAPK pathway is highly responsive to light input during the subjective night and insensitive to light during the middle of the subjective day. These results provide insight into the molecular mechanisms underlying the effects of photoperiod on the immune regulation of E. sinensis.

The Biochemical Composition and Quality of Adult Chinese Mitten Crab Eriocheir sinensis Reared in Carbonate-Alkalinity Water

Saline–alkaline aquaculture has become an important breakthrough in expanding the space available for aquaculture in China. However, the biochemical composition and quality of Eriocheir sinensis reared in carbonate alkalinity water are still unclear. Therefore, this study investigated the edible yield, coloration, and nutritional and flavor quality of Eriocheir sinensis. A significantly lower gonadosomatic index (GSI), meat yield (MY), and total edible yield (TEY) were detected in intensive pond (IP) samples than those in semi-intensive reed wetland (SIWR) (p &lt; 0.05). Six color parameters in the hepatopancreas (p &lt; 0.05) differed between IP and SIRW. The contents of crude protein and fat in the female hepatopancreas of IP were significantly higher than those in SIRW (p &lt; 0.05). The concentrations of total monounsaturated fatty acids (∑MUFA), total essential fatty acids (∑EFA), and hypocholesterolaemic/hypercholesterolaemic ratio (h/H) in the female edible tissues checked were higher in IP than those in SIRW, with significant differences including ∑MUFA in the hepatopancreas and ovary, ∑EFA in the muscle, and h/H in the ovary (p &lt; 0.05). Higher total free amino acid (∑FAA) contents of muscle were detected in SIRW than that in IP samples. Significantly higher K, Ca, Mg, Fe, and Zn contents in the ovary were detected in SIRW samples compared to IP (p &lt; 0.05). Overall, Eriocheir sinensis reared in carbonate-alkalinity water is an important source of nutrients.