Crab Portunus Trituberculatus Research Articles

Elucidating the role of DNA methylation in low-salinity adaptation of the swimming crab Portunus trituberculatus

The swimming crab Portunus trituberculatus holds significant economic value in the aquaculture industry in China. Salinity is an important environmental factor that affects its growth and susceptibility to diseases. DNA methylation, a well-studied epigenetic mechanism, plays a pivotal role in the stress response of organisms. Herein we performed whole-genome bisulfite sequencing (WGBS) to assess changes in DNA methylation patterns in P. trituberculatus gill tissues before and after exposure to low-salinity stress. Overall, 33,712 differentially methylated regions (DMRs) were screened and 3546 functional genes were identified, many of which were notably enriched in pathways related to, for example, proteolysis and amino acid metabolism. By integrating WGBS and previous RNA-seq data, we identified 187 overlapping genes between differentially expressed genes and differentially methylated genes. Our findings reveal molecular mechanisms via which DNA methylation participates in low-salinity adaptation by regulating gene expression. Moreover, our data provide preliminary evidence of the substantial role of DNA methylation in facilitating salinity adaptation in P. trituberculatus and contribute to a systematic understanding of the mechanisms underpinning low-salinity adaptation in this species.

Temporal and tissue-specific regulation of energy metabolism in the swimming crab Portunus trituberculatus during nitrite stress and recovery

Nitrite is a common pollutant in aquaculture systems that poses a significant threat to aquatic animals. Energy metabolism is critical in ensuring survival of animals under environmental stressors. However, regulation of energy metabolism in crustaceans under nitrite stress has not been well understood. Here we investigated energy metabolism regulation during nitrite stress and recovery in different tissues of the swimming crab Portunus trituberculatus, an important aquaculture species in China. Our results revealed that nitrite can cause tissue hypoxia and impair energy homeostasis, and energy balance cannot be restored even after a 96-hour recovery. Following exposure, mobilization of glycogen and lipids exhibited different temporal patterns. In response to energy imbalance, AMPK signaling was activated to counter energy imbalance. However, prolonged nitrite stress impaired AMPK signaling, leading to a further decline in energy supply. The findings improve our understanding for nitrite toxicity in P. trituberculatus, and provide valuable information for aquaculture management.

Dietary supplementation of Roseovarius litoreus improves survival and immune defense of the swimming crab Portunus trituberculatus

The aquaculture of the swimming crab Portunus trituberculatus is challenged by vibriosis, and Rhodobacteraceae strains play a crucial role in maintaining gut health and enhancing the immunity of crustaceans. In this study, we investigated the effects of dietary supplementation of Roseovarius litoreus, a member of Rhodobacteraceae, on the survival and immune defense of the swimming crab. The results showed that 60-day dietary supplementation of R. litoreus resulted in a marked increase in the mRNA expression of gut ZO-1. Meanwhile, R. litoreus increased zero-radius operational taxonomic unit (ZOTU) richness and changed the gut bacterial community, which was manifested by the enriched Carboxylicivirga and Roseovarius and depleted Photobacterium and Vibrio, thereby upregulating functional potentials of the gut bacterial community in the immune system and biosynthesis of other secondary metabolites. After a 72-h V. parahaemolyticus challenge, R. litoreus-supplemented crabs had an increased survival rate, reduced histological damage of hepatopancreas and gut, and improved expressions of immune-related genes and lysozyme activity. Moreover, R. litoreus-supplemented crab gut still harbored more Carboxylicivirga and Roseovarius but had fewer Vibrio gene copies than control crabs, which led to similar changes in functional potentials. Notably, all ZOTUs that were positively correlated with immune-related genes and lysozyme activity belonged to Bacteroidetes, Bacteroidales, or Carboxylicivirga. Taken together, we highlight the probiotic role of R. litoreus in the survival and immune defense of the swimming crab. These findings may contribute to the development of indigenous bacteria of the swimming crab as potential probiotics for the control of vibriosis.

Insights into lipid function for ovarian development in the swimming crab (Portunus trituberculatus): A comparison of lipid mobilization and deposition in hepatopancreas and ovary

Ovarian development is a complex and dynamical process regulated by the coordinated actions of the hepatopancreas and ovary. In mammals, lipids provide necessary energy for oocytes to mature, whereas their regulatory role in crustaceans is not fully understood. This study compares lipid mobilization and deposition in hepatopancreas and ovary of the swimming crab at different stages of ovarian maturation. The results revealed that cholesterol (CHO) and triglyceride (TG) content in hepatopancreas increased with ovary development increased from stage I to Ⅲ, and then decreased at stage V, while those content in ovary significantly increased from stage I to V. Additionally, apolipoprotein (ApoA and ApoB) contents in hepatopancreas and ovary increased significantly during ovary development. Moreover, the highest expression levels of cholesterol transport related genes (srb, ldlr, lrp2 and abcg1) were observed in the hepatopancreas at stage Ⅲ and in the ovary at stage Ⅳ. Furthermore, as ovary development form stage I to Ⅳ, the hepatopancreas up-regulated the expression levels of genes related to lipid anabolism (fas, acc, g6pd, and 6pgd) and fatty acid transport (fabp9 and fatp4). Concurrently, genes related to lipid catabolism (il) and fatty acid β-oxidation (acox2 and acox3) in the hepatopancreas were down-regulated. Differently, with the ovary development increased from stage I to Ⅳ, the ovary up-regulated gene expression levels involved in lipid anabolism (acc, 6pgd, g6pd, dgat1, and gpat3), fatty acid synthesis (elovl and elovl4) and fatty acid β-oxidation (cpt1, cpt2, acox1, and acox3). The mRNA expression levels of lipolysis (tgl) and fatty acid transport (fabp1, fbap9, and fatp4) in ovary were down-regulated. Overall, these findings demonstrate that the hepatopancreas and ovary exhibit different responses to lipid metabolism during ovarian development.

Valine biased glycogen consumption to promote energy metabolism of Portunus trituberculatus and increased protein and essential amino acid deposition in muscle

The present research assessed the impact of dietary valine on nutritional and energy metabolism, as well as muscle development in swimming crabs (Portunus trituberculatus) over a period of six weeks. Swimming crabs, initially weighing around 23.34 ± 0.03 g, were given six diets identical in nitrogen and lipid levels (45 % and 8 % respectively). These diets contained valine contents of 0.99 %, 1.24 %, 1.56 %, 1.69 %, 1.90 %, and 2.20 % respectively. One hundred and eighty crabs were randomized into six treatments, each treatment including three replicates. The results suggested that significant increases were discovered in FW, PWG, as well as SGR of crabs when dietary valine increasing from 0.99 % to 1.69 %, but these measurements decreased when dietary valine increased to 2.20 %. Compare to the control treatment, crabs that were given diets with 1.90 % and 2.20 % valine showed increased level of muscle protein. Crabs that were given the diets containing 1.56 % and 1.69 % valine showed significantly increased concentration of GLU in hemolymph compared to the control group. Additionally, crabs that were given the diets containing 1.69 % valine and 1.90 % valine respectively showed significantly increased concentrations of T-CHO and TG in hepatopancreas. Hepatic glycogen concentration significantly decreased as the level of valine in diets increased from 0.99 % to 1.56 %, but significantly increased as valine level further increased to 2.20 %. Compared with the control group, crabs that were given diets with 1.90 % valine showed increased expression levels of genes associated to amino acid metabolism (pi3k, akt, alt, ast, 4ebp1 and eif4e3) and energy metabolism (pk, cs1, idh, nd1, cytb and cox2). Crabs given diets containing 2.20 % valine showed significantly increased expression level of pepck in hepatopancreas, while crabs given diets containing 1.24 %, 1.56 %, and 1.69 % valine exhibited significantly increased expression level of cpt1 in hepatopancreas. Crabs given the diet with 1.69 % valine had the highest expression levels of tor and pax7 in muscle. Increased dietary valine significantly reduced myofiber density but promoted higher frequency of myofibers in larger diameters. In conclusion, the optimal valine requirement for P. trituberculatus was 1.56 % of diet based on PWG by a quadratic modeling. 1.69–1.90 % valine promoted amino acid metabolism and muscle growth, moreover, enhanced energy metabolism by driving the consumption of glycogen rather than lipids.

Bacterial dynamics and biotic sources in the developing swimming crab embryos

The microbial community associated with aquatic animals begins to assemble from the embryonic stage and plays a vital role in their growth and survival. However, the microbial community of the swimming crab Portunus trituberculatus during its embryonic period has not yet been studied. In this study, we investigated the embryonic bacterial community (EBC) of this species across the developmental cycle to elucidate its longitudinal dynamics and biotic sources. The results showed that the diversity of the EBC decreased with host development. The composition of the EBC varied across embryonic development, with a high percent of persistent bacteria and the gradual replacement of γ-Proteobacteria (mainly Oceanospirillaceae and Colwelliaceae) by α-Proteobacteria (Rhodobacteraceae and Flavobacteriaceae) as the major signatures of the succession patterns. Moreover, the unique amplicon sequence variants (ASVs) exhibited the highest functional potentials at the cleavage stage, whereas the shared ASVs did so at the later stages. Furthermore, maternal tissue, mainly the ovary and abdominal setae, contributed the most to the EBC, while rearing water contributed minimally. Overall, our results reveal succession patterns and biotic sources of EBC during the developmental cycle, highlighting that the dominance of embryonic bacteria starts from the early stage of embryo and bacterial vertical transmission is of significance during embryonic development. We also suggest the necessity of microbial management from maternal crabs.

Vitamin D3 alleviates the damage caused by high-fat and maintains lipid metabolism homeostasis in hepatopancreas by promoting fatty acid β-oxidation of Portunus trituberculatus

The purpose of present study was to evaluate the effects of vitamin D3 supplementation in high-fat diet on growth performance, calcium and phosphorus in tissue, immunity and lipid metabolism, and to explore whether adding vitamin D3 to high-fat diet can alleviate the adverse effects of high-fat diet on swimming crab (Portunus trituberculatus). The three experimental diets were normal fat diet (NFD, 9.35 % fat, 45.53 % protein), high fat diet (HFD, 14.69 % fat, 44.93 % protein) and high fat diet supplemented with 0.47 mg/kg VD3 (HFD+VD), respectively. A total of 108 healthy swimming crabs with an initial body weight of 9.42±0.13 g were selected for an 8-week feeding trial. The results showed that crabs fed with HFD+VD diet exhibited significantly higher percent weight gain (PWG), specific growth rate (SGR), feed efficiency (FE) and molting rate (MR) compared to those fed with HFD diet (P < 0.05). It was observed from hepatopancreas crude lipid content, TG content and hepatopancreas sections that HFD caused hepatopancreatic cell damage and abnormal lipid accumulation (P < 0.05), however, there were no significant differences in hepatopancreas crude fat content, TG content and hepatopancreas section observation between HFD+VD group and NFD group (P > 0.05). Crabs fed HFD diets with or without VD3 significantly down-regulated the expression of genes related to fatty acid transport, such as fabp9 and lpr1 in hepatopancreas (P < 0.05), moreover, the expression of genes related to fatty acid β-oxidation (cpt2 and acox1) were significantly up-regulated and genes related to lipid synthesis (fas and 6pgd) were significantly down-regulated in the hepatopancreas of the crabs fed with HFD+VD diet compared to those fed with HFD diet (P < 0.05). In conclusion, the results of present study indicated that a high-fat diet can cause deformation and rupture of hepatopancreas cells through abnormal lipid accumulation, thereby causing associated inflammation, leading to large calcium and phosphorus deposits in hemolymph and muscle, however, high fat diet supplemented with vitamin VD3 mainly enhanced the β-oxidation of fatty acids, inhibited lipid synthesis, alleviated abnormal lipid accumulation, and thus maintained the balance of lipid metabolism in hepatopancreas cells and promoted growth performance and feed utilization for swimming crab.

Physiological and Transcriptomic Analyses Provide Insights into Nitrite Stress Responses of the Swimming Crab Portunus trituberculatus.

Nitrite is a common environmental pollutant in intensive aquaculture systems. In this study, physiological and transcriptomic analyses were performed to investigate nitrite stress responses in the swimming crab Portunus trituberculatus, an important aquaculture species in China. The results revealed that nitrite can affect neurotransmitter signaling via the expression of neurotransmitter receptors such as octopamine receptor (OAR) and 5-hydroxytryptamine receptor (5-HTR), and depress ecdysteroid signaling by downregulating ecdysteroid receptor (EcR) as well as its downstream transcription factors in hepatopancreas. In addition, nitrite suppressed the expression of hemocyanins, the oxygen-transporting protein, which at least partly contributed to tissue hypoxia, resulting in a switchover of energy metabolism from aerobic to anaerobic pathway. To meet the energy demand, glycogens and lipids were mobilized and transported to the hemolymph, and the catabolism of amino acids and fatty acids was enhanced to provide energy for hepatopancreas. β-oxidation of fatty acids, the major process by which fatty acids are oxidized to generate energy, seems to occur mainly not in mitochondria but in peroxisomes. Although the cellular protective mechanisms, including antioxidant defense, heat shock response (HSR), unfolded protein response (UPR), and autophagy, were activated, nitrite-induced cellular stress overwhelmed the repairing capacity and caused significant increase in the levels of apoptosis. These results indicated that nitrite stress influences neurotransmitter and endocrine signaling, disturbs energy metabolism, damages cellular components, and induces apoptosis in P. trituberculatus. The findings of this study provide new insights into nitrite stress response in the swimming crab and provide valuable information for aquaculture management of this species.

Hitchhiking on drifting seaweed reduces predation risk in juveniles of the swimming crab Portunus trituberculatus

Hitchhiking on drifting seaweed reduces predation risk in juveniles of the swimming crab Portunus trituberculatus

Temporal dynamics of microbial communities in the water of polyculture pond system for Chinese swimming crab Portunus trituberculatus

Microorganisms as the vital component of aquaculture ecosystems, were involved in nutrient cycling, water quality regulation, and the health of cultured species. In this study, aquatic water samples were collected monthly to reveal microbial community compositions, functions, and interactions in different habitats (pond and waterway) during the culture stages of the Chinese swimming crab Portunus trituberculatus in Shandong Province. Due to the semi-closed conditions with limited water exchange, the α diversities were lower in the polyculture pond. The relative abundances of the dominant phyla varied dramatically, and most biomarkers were identified in the polyculture pond. The β diversity of the microbial communities significantly differed among culture stages and habitats. Furthermore, the heterogeneity of prokaryotic microorganisms was greater than that of eukaryotic microorganisms. In the polyculture pond, the microbial community was sensitive to nutrient cycling, with abundant functions related to “photoautotrophy”, “nitrogen_respiration” and “nitrate_respiration”, corresponding to P. trituberculatus culture activities. The co-occurrence network of the microbial community in the polyculture pond tended to be looser than that in the waterway, with a lower number of taxa and correlations. In addition, these networks had nonrandom patterns and formed “small world” topological structures. The low-abundance genera with a higher level of betweenness centrality scores were the hubs or connectors in the networks and might play an essential role in microbial community composition. Furthermore, potential pathogens (Vibrio, Acholeplasma, Photobacterium, and Flavobacterium) deserve more attention during P. trituberculatus culture. This study will be useful for understanding microbial communities in polyculture pond systems and will provide a reference for the health aquaculture of P. trituberculatus.

Machine learning-based aggressiveness assessment model construction for crabs: A case study of swimming crab Portunus trituberculatus

Aggressiveness trait-based selection is crucial for alleviating interspecies cannibalism in economic crab species and enhancing survival rates in aquaculture. However, there is a lack of efficient and simple methods for assessing aggressiveness. In this study, we measured aggressiveness of the swimming crab Portunus trituberculatus through repeated mirror tests and fighting experiments. Factor analysis and the K-means algorithm were used to assess aggressiveness quantitatively and qualitatively. A combination of multiple linear regression and support vector machine (SVM) analyses was employed to construct an aggressiveness assessment model for swimming crabs and explore the relationship between aggressiveness and fighting ability. The results showed significant correlations among repeated aggressive behaviors (attacking, chela extending, defending, crossing, reverse walking, and freezing). Aggression score was significantly correlated with fighting behaviors, and there were significant differences in fighting abilities among different levels of aggressiveness. This suggested that aggressive behaviors are consistent within individuals and that aggressiveness, as a personal trait, affects the fighting ability of swimming crabs. Aggression score (Y) and clustering results of K-means can serve as assessment indicators of aggressiveness. The predictive variables for the quantitative assessment model were relative movement distance (X1) and freezing duration (X2). The adjusted R-square of the optimized quantitative model was 0.72, it also had the smallest Sigma, AIC, MSE, and RMSE values and the best fitting regression equation, which was Y = 0.023X1 – 0.001X2 – 0.002. The predictor variables for the qualitative assessment model were relative movement distance, freezing frequency, and duration. SVM was used to construct the qualitative model, and the prediction accuracy was 92%, sensitivity was 84%, and specificity was 100%, indicating the model has a good classification and prediction effect. The machine learning-based aggressiveness assessment model constructed in this study provides a behavioral method for the selection and high-throughput measurement of economic crab species with excellent aggressiveness traits, giving it important industrial application value.

Identification of Two Insulin Receptors from the Swimming Crab Portunus trituberculatus: Molecular Characterization, Expression Analysis, and Interactions with Insulin-Like Androgenic Gland Hormone.

AbstractThe insulin-like androgenic gland hormone is a crucial sexual regulator that is involved in the masculine sexual differentiation of crustaceans. As an insulin-like peptide, the insulin-like androgenic gland hormone has been proposed to act through the insulin receptor-mediated pathway. The present study cloned and characterized two insulin receptors (PtIR1 and PtIR2) from the swimming crab Portunus trituberculatus hallmarked with a conserved intracellular tyrosine kinase catalytic domain and several other typical insulin receptor domains in their deduced amino acid sequences. Both insulin receptors were predominately expressed in the testis and the insulin-like androgenic gland hormone-producing organ androgenic gland. Their testicular expression during the annual cycle suggested that they may play critical roles in spermatogenesis. By using the protein colocalization analysis in HEK293 cells, interactions of PtIAG with the two PtIRs were further confirmed. In addition, the insulin receptor antagonist was found to attenuate the stimulatory effects of androgenic gland homogenate on the phosphorylated MAPK levels in testis explants, suggesting that the insulin receptor-dependent MAPK pathway may be essential for insulin-like androgenic gland hormone functions.

Effect of dietary β‐carotene supplementation on growth and antioxidant capability of female swimming crab Portunus trituberculatus

AbstractDietary supplementation with synthetic β‐carotene was investigated in this study to assess its impact on gonad development, antioxidant status, color parameters, and carotenoid composition in adult female swimming crab (Portunus trituberculatus). Four diets, designated as Diet 1 to 4, were formulated to contain 3.07, 53.78, 103.23, and 205.22 mg/kg dry diets of synthetic β‐carotene. The findings revealed the following: (1) No significant differences in body weights or feed utilization were observed, indicating that β‐carotene did not significantly enhance the growth of adult P. trituberculatus. (2) The inclusion of 50–100 mg/kg β‐carotene in the diets led to improvements in antioxidant capacity indicators (T‐AOC, MDA) and immune‐related parameters (AKP, ACP) in both hepatopancreas and hemolymph. (3) Notably, the redness of ovaries and hepatopancreas increased significantly with higher dietary β‐carotene supplementation for color enhancement. In conclusion, incorporating β‐carotene into the diet enhanced the antioxidative capability, nonspecific immunity, and tissue carotenoid contents in adult female P. trituberculatus, and the optimal level of dietary synthetic β‐carotene supplementation for adult female P. trituberculatus was suggested to be around 100 mg/kg.

The impact of noise on the stress response of Portunus trituberculatus in a land-based monoculture system

Human-induced noise, particularly from shipping and construction, significantly affects the stress responses of swimming crabs (Portunus trituberculatus Miers, 1876). Prolonged exposure can lead to stress responses, impacting the overall health of the crabs. This study aimed to assess how noise affects stress responses in P. trituberculatus. The sound had an intensity of 163 dB 1μPa for the noise condition and 114 dB 1μPa for ambient sound. P. trituberculatus was continuously exposed to the noise stimulus for six hours. Samples of serum, muscle tissue, gills, and hepatopancreas were collected at multiple time points: 0 min, 10 min, 30 min, 1 h, 3 h, 6 h, and 7 h. The serum cortisol levels in the noise-exposed group were significantly higher than those in the control group at 30 min and 6 h. The two groups had no significant differences regarding serum glucose and muscle lactic acid content. In the noise-exposed group, the gills exhibited significantly lower superoxide dismutase (SOD) levels. Conversely, for the hepatopancreas, the SOD levels in the noise-exposed group were notably lower than those in the control group at 10 min but significantly higher at 6 h and 7 h. Malondialdehyde (MDA) levels were substantially higher in the noise-exposed group than in the control group at 30 min and 6 h. This study demonstrates that P. trituberculatus experiences a physiological stress response to acoustic sounds.

Reassembly and biotic sources of carapace bacterial community of Portunus trituberculatus after host molting

Crustaceans molt repeatedly during their development, which leads to frequent reassembly of their associated microbial communities. However, the reassembly of the carapace bacterial community (CBC) and its sources remain largely unclear. Here, these concerns were explored by investigating the succession pattern, assembly processes, and biotic sources of CBC of the swimming crab Portunus trituberculatus after host molting. Rhodobacteraceae, significantly discarded with the molted hard shell, was recruited during the 72 h postmolt. The early colonists such as Pseudoalteromonadaceae and Oeanospirillaceae had the higher physiological functional potentials for colonization, mainly via homogeneous selection. By contrast, the late colonists such as Flavobacteriaceae and Pseudomonadaceae had higher metabolic potentials and their colonization was governed by drift first and dispersal limitation later. The CBC was primarily shaped by multiple colonist pools, especially preceding carapace bacteria. Seawater had a higher contribution for CBC at the early stage than the late stage via providing the key bacteria mainly Rhodobacteraceae. These findings advance our understanding of the CBC and its reassembly pattern and sources, highlighting the importance of seawater bacteria for the early colonists. We also suggest the possibility and potential timing of using probiotics in the rearing water for the management of carapace microbiota in the aquaculture practice.

Selectivity of crab pots with escape vents and the optimal vent height for reducing the catch of sublegal swimming crab in the East China Sea pot fishery

Catching and subsequent discarding of sublegal-sized swimming crab Portunus trituberculatus in the pot fishery may result in increased unaccounted fishing mortality and contribute to overexploitation of the crab stocks in the East China Sea. Developing technical measures and implementing related regulations to improve the size selection of pots to reduce the catch of sublegal crab has become urgent. In this study, we evaluated the size selectivity of pots with rectangular escape vents of different opening heights (25 mm, 28 mm, 30 mm and 33 mm) through comparative fishing experiments and by employing the SELECT model. The results show that the installation of escape vents significantly reduced the catch per pot (in numbers) of total and sublegal swimming crabs, and significantly increased the proportion of legal-sized crabs for most vented pots except the 25 mm vent height. A significant positive linear relationship was found between escape vent height and the 50% retention carapace width, selection range and fishing power. Increasing the height of the escape vent had substantial effect on the exploitation pattern for swimming crab and the pot with a 28 mm height escape vent seemed to be the optimal design, releasing 67.4% of sublegal-sized crab and maintaining 94.8% of legal-sized crabs. Results showing that a pot with an escape vent that significantly reduces sublegal crab catch, while maintaining catch efficiency for legal-sized crabs, is positive for the fishery and supports the implementation of escape vents in the East China crab pot fishery.

Evaluation of spray-dried blood meal for application in commercial-like feed for juvenile swimming crab (Portunus trituberculatus)

The objective of the present study was to evaluate the effects of replacing fish meal (FM) with spray-dried blood meal (SDBM) on growth performance, antioxidant capacity, digestive enzyme activity, tissue Fe content, gene expression related to inflammation, and the TOR pathway in juvenile swimming crab (Portunus trituberculatus). Four isonitrogenous (47.0% protein) and isolipidic (9.0% lipid) diets were formulated to contain different levels of SDBM, with FM replaced by SDBM at 0, 10%, 20%, and 30%, respectively. The results showed that crabs fed diet SDBM30 exhibited the lowest survival among all treatments, and crabs fed the SDBM20 diet had the highest final weight, percent weight gain, specific growth rate and feed efficiency among all treatments. In addition, the protein content of the crab hepatopancreas decreased significantly and the lipid content increased significantly as SDBM replaced FM in the diets. Compared to other diets, crabs fed the SDBM30 diet had the lowest activities of antioxidant enzymes and the highest concentration of malondialdehyde in the hepatopancreas and hemolymph. The Fe contents in hepatopancreas and gill increased significantly, and the color of the hepatopancreas got darker as replacement of FM with SDBM increased from 0% to 30%. The expression of genes related to TOR (tor, akt, s6, and s6k1) and antioxidant (gpx) pathways in the hepatopancreas were down-regulated significantly with increased replacement of dietary FM with SDBM. In contrast, expression levels of genes related to protein translation (4ebp1, eif4e1a, eif4e2, and eif4e3) and inflammation pathways (myd88, traf6, relish, and irak4) were up-regulated significantly in crabs fed diets with SDBM supplementation. In summary, replacing FM with less than 20% of SDBM can promote the growth, but can darken the hepatopancreas, negatively affect antioxidant capacity, and induce an inflammatory response.

Combined effects of pentachlorophenol and nano-TiO2 with different sizes on antioxidant, digestive, and immune responses of the swimming crab Portunus trituberculatus

Marine nano-titanium dioxide (nano-TiO2) and pentachlorophenol (PCP) pollution are escalating concerns in coastal areas. This study investigated the combined effects of continuous exposure to nano-TiO2 (25 nm, 100 nm) and PCP (0, 1, 10 μg/L) for 28 days on the antioxidant, digestive, and immune abilities of the swimming crab Portunus trituberculatus. Compared with the control group, the interaction between nano-TiO2 and PCP was significantly higher than exposure to a single stressor, with a pronounced decrease in amylase activity observed due to the reducing nano-TiO2 particle sizes. Resulting in increased MDA and SOD activity. The expression levels of Toll4, CSP3, and SER genes in crab hemolymph showed perturbations following exposure to nano-TiO2 and PCP. In summary, according to the results of CAT, GPX, PES and AMS enzyme activities, it was concluded that compared to the larger particle size (100 nm), the single stress of nano-TiO2 at a smaller particle size (25 nm) and co-stress with PCP have more significant impacts on P. trituberculatus. However, the potential physiological regulation mechanism of the interaction between these pollutants remains elusive and requires further study.

Protein and lipid nutrition in crabs

AbstractUnderstanding the nutrition of crabs has a key role in ensuring the success and sustainability of their culture, as providing a well‐balanced, cost‐effective and sustainable diet that ensures the survival, growth and health of crabs is crucial. The present review is the first to focus primarily on the current state of knowledge of the nutrient requirements and related nutritional aspects in farmed crab species. The most common farmed and studied crabs are the Chinese mitten crab (Eriocheir sinensis Milne‐Edwards, 1853), giant mud crab (Scylla serrata Forsskal, 1775), green mud crab (Scylla paramamosain Estampador, 1949) and swimming crab (Portunus trituberculatus Miers, 1876). The article reviews how levels of dietary protein and lipid, the two most important and expensive macronutrient ingredients for most marine animals, directly affect reproduction, growth performance and survival of crabs, and the important impacts they have on immune response and antioxidant capacity. Furthermore, essential amino acids, and essential fatty acids, especially the long‐chain polyunsaturated fatty acids, as well as key lipid classes such as cholesterol and phospholipid will be discussed in terms of dietary requirements. Nutrient digestibility is a crucial method to determine protein quality, and studies on this topic in crabs were covered. The replacement of fishmeal and fish oil, as the predominant ingredients traditionally used in aquafeeds for marine animals, with more sustainable alternatives in diet formulations for crabs are also discussed. Modern ‘omics’ studies and high‐throughput technologies as fast‐growing approaches in protein and lipid research are also covered. Crabs generally require approximately 35%–50% protein, 5%–10% lipid, ~2.5% arginine, ~2.5% lysine, 1.5%–2.5% phenylalanine, 2.2% leucine, 0.7% tryptophan, 0.7% taurine, 1%–2% each of eicosapentaenoic acid, docosahexaenoic acid and arachidonic acid, 1% cholesterol and ~2% phospholipid in their diets.

Overcoming research challenges: In vitro cultivation of Ameson portunus (Phylum Microsporidia)

Ameson portunus is an intracellular pathogen that infects marine crabs Portunus trituberculatus and Scylla paramamosain, causing significant economic losses. However, research into this important parasite has been limited due to the absence of an in vitro culture system. To address this challenge, we developed an in vitro cultivation model of A. portunus using RK13 cell line in this study. The fluorescent labeling assay indicated a high infection rate (∼60 %) on the first day post-infection and quantitative PCR (qPCR) detection demonstrated successful infection as early as six hours post-inoculation. Fluorescence in situ hybridization (FISH) and qPCR were used for the detection of A. portunus infected cells. The FISH probe we designed allowed detection of A. portunus in infected cells and qPCR assay provided accurate quantification of A. portunus in the samples. Transmission electron microscopy (TEM) images revealed that A. portunus could complete its entire life cycle and produce mature spores in RK13 cells. Additionally, we have identified novel life cycle characteristics during the development of A. portunus in RK 13 cells using TEM. These findings contribute to our understanding of new life cycle pathways of A. portunus. The establishment of an in vitro culture model for A. portunus is critical as it provides a valuable tool for understanding the molecular and immunological events that occur during infection. Furthermore, it will facilitate the development of effective treatment strategies for this intracellular pathogen.