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The eggshell color of avian species is an important trait that is predominantly determined by the pigments biliverdin and protoporphyrin. Various factors affect eggshell pigment deposition and coloration; however, the underlying mechanisms remain unclear. We analyzed the hepatic transcriptomes and metabolomes of Changshun green-shell hens laying dark green and light green eggs to investigate the potential role of the liver in regulating the intensity of the green eggshell color. In total, 350 differentially expressed genes and 211 differentially altered metabolites were identified. Gene set enrichment analysis revealed that the enriched pathways and Gene Ontology (GO) terms were mainly associated with energy, immunity, and nutrient metabolism. Metabolite set enrichment analysis revealed that the enriched pathways were mainly associated with amino acid, vitamin, bile acid, and lipid metabolism. Moreover, gene-metabolite interaction network analysis revealed 1 subnetwork. Most genes and metabolites in this subnetwork were determined to be related to melanin metabolism and transport. In conclusion, our results suggest that hepatic melanin metabolism and transport are critical for eggshell coloration. Six candidate genes (CDKN2B, DDC, PYCR1, ABCG5, SLC3A1, and P2RX2) and 7 candidate metabolites (serotonin, 5-hydroxyindoleacetic acid, ornithine, acetylcholine, L-tryptophan, D-ornithine, and ADP) were suggested to play important roles in this process. Meanwhile, this study suggests that changes in hepatic energy metabolism, immune status, antioxidation activity, nutrient availability, and bile acid synthesis can impair eggshell coloration., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Image anomaly detection is a trending research topic in computer vision. The objective is to build models using available normal samples to detect various abnormal images without depending on real abnormal samples. It has high research significance and value for applications in the detection of defects in product appearance, medical image analysis, hyperspectral image processing, and other fields. This paper proposes an image anomaly detection algorithm based on feature distillation and an autoencoder structure, which uses the feature distillation structure of a dual-teacher network to train the encoder, thus suppressing the reconstruction of abnormal regions. This system also introduces an attention mechanism to highlight the detection objects, achieving effective detection of different defects in product appearance. In addition, this paper proposes a method for anomaly evaluation based on patch similarity that calculates the difference between the reconstructed image and the input image according to different regions of the image, thus improving the sensitivity and accuracy of the anomaly score. This paper conducts experiments on several datasets, and the results show that the proposed algorithm has superior performance in image anomaly detection. It achieves 98.8% average AUC on the SMDC-DET dataset and 98.9% average AUC on the MVTec-AD dataset.

An 8-week feeding trial was conducted in Pacific white shrimp ( Litopenaeus vannamei ) to evaluate the effects of dietary choline supplementation on choline transport and metabolism, hepatopancreas histological structure and fatty acid profile, and regulation of lipid metabolism. Six isonitrogenous and isolipidic diets were formulated to contain different choline levels of 2.91 (basal diet), 3.85, 4.67, 6.55, 10.70 and 18.90 g/kg, respectively. A total of 960 shrimp (initial weight, 1.38 ± 0.01 g) were distributed randomly into twenty-four 250-L cylindrical fiber-glass tanks, with each diet assigned randomly to 4 replicate tanks. The results indicated that dietary choline significantly promoted the deposition of choline, betaine and carnitine ( P  < 0.05). The diameters and areas of R cells, total lipid and triglyceride contents in hepatopancreas, and triglyceride and non-esterified fatty acid contents in hemolymph were negatively correlated with dietary choline level. The contents of functional fatty acids in hepatopancreas, the activity of acetyl-CoA carboxylase (Acc), and the mRNA expression of fas , srebp and acc were highest in shrimp fed the diet containing 4.67 g/kg choline, and significantly higher than those fed the diet containing 2.91 g/kg, the lowest level of choline ( P  < 0.05). The number of R cells, content of very low-density lipoprotein (VLDL), activities of carnitine palmitoyl-transferase (Cpt1), lipoprotein lipase and hepatic lipase, and the mRNA expression levels of cpt1 , fabp , fatp , ldlr , and ampk in hepatopancreas increased significantly as dietary choline increased ( P  < 0.05). In addition, hepatopancreas mRNA expression levels of ctl1 , ctl2 , oct1 , badh , bhmt , ck , cept , and cct were generally up-regulated as dietary choline level increased ( P  < 0.01). In conclusion, dietary choline promoted the deposition of choline and its metabolites by up-regulating genes related to choline transport and metabolism. Moreover, appropriate dietary choline level promoted the development of hepatopancreas R cells and maintained the normal accumulation of lipids required for development, while high dietary choline not only promoted hepatopancreas lipid export by enhancing VLDL synthesis, but also promoted fatty acid β-oxidation and inhibited de novo fatty acid synthesis by activating the Ampk/Srebp signaling pathway. These findings provided further insight and understanding of the mechanisms by which dietary choline regulated lipid metabolism in L. vannamei ., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2023 The Authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd.)

The objective of the present study was to evaluate the effects of dietary choline levels on growth performance, antioxidant capacity, innate immunity and hemocyte apoptosis of Litopenaeus vannamei. Six isonitrogenous and isolipidic diets were formulated to contain different choline levels: 2.91 (basal diet), 3.85, 4.67, 6.55, 10.70 and 18.90 g kg -1 choline, respectively. The results indicated that shrimp fed diet with 4.67 g kg -1 choline had the highest final body weight (FBW), percent weight gain (PWG), specific growth rate (SGR), feed efficiency (FE), and activities of alkaline phosphatase (AKP) and phenoloxidase (PO) in hemolymph among all treatments. Shrimp fed diet with 18.90 g kg -1 choline exhibited significantly lower crude lipid in hepatopancreas than those fed diets with 2.91, 3.85, 4.67 and 6.55 g kg -1 choline (P < 0.05). The concentration of reactive oxygen species (ROS) and apoptosis rate in hemocytes significantly decreased with the increase of dietary choline levels (P < 0.05). Shrimp fed diets with 6.55, 10.70 and 18.90 g kg -1 choline had significantly higher scavenging ability of hydroxyl radical (SAHR) and total antioxidant capacity (T-AOC) in hemolymph than those fed diet with 2.91 g kg -1 choline (P < 0.05). Dietary choline supplementation down-regulated the expression of genes related to apoptosis such as caspase-1, caspase-3, caspase-8, p53, and p38MAPK in hemocytes (P < 0.05), while up-regulated the expression of anti-apoptosis gene bcl2 in hemocytes (P < 0.05). Overall, the results of the present study demonstrated that appropriate dietary choline could improve growth performance and feed utilization, enhance antioxidant capacity and innate immunity, and mitigate apoptosis in Litopenaeus vannamei. Moreover, the inhibition of hemocyte apoptosis by dietary choline may be regulated by the p38MAPK-p53 signaling pathway., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

An eight-week feeding trail was carried out to investigate the impacts of different dietary arachidonic acid (ARA) supplementations on growth performance, antioxidant capacity, tissue fatty acid profiles, and lipid metabolism of mud crab ( Scylla paramamosain ) juvenile. Six isonitrogenous (480 g kg -1 crude protein) and isolipidic (80 g kg -1 crude lipid) diets were formulated to contain 0.40, 2.50, 4.60, 8.90, 12.50, and 15.70 g ARA kg -1 (dry matter), respectively. Each experimental treatment included 24 mud crab juveniles (initial weight 11.29 ± 0.09 g) and was assigned to triplicate groups ( n = 3). Crabs fed diets with 2.50, 4.60, and 8.90 g kg -1 ARA presented significantly higher percent weight gain (PWG) and specific growth rate (SGR) than those fed the other diets. Based on two-slope broken-line and quadratic curve regression analysis of PWG against dietary ARA levels, optimal dietary ARA levels were determined to be 5.20 g kg -1 and 6.20 g kg -1 , respectively. Crabs fed with 4.60 g kg -1 ARA diet showed the lowest activities of alanine aminotransferase (ALT) as well as aspartate aminotransferase (AST) in hemolymph among all treatments. In hemolymph and hepatopancreas, total antioxidant capacity (T-AOC), the activities of total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px) as well as the contents of reduced glutathione (GSH) rose first and then dropped with the increase of dietary ARA levels, while the concentration of malondialdehyde (MDA) showed an opposite trend. Tissue fatty acid profiles reflected diets fatty acid compositions. The ARA contents in hepatopancreas and muscle significantly increased with the increase of dietary ARA levels. Furthermore, the areas of blasenzellen (B) cells and restzellen (R) cells were significantly downregulated with the increase of dietary ARA levels. Crabs fed with 0.40 g kg -1 ARA diet showed significantly higher gene expression levels of fatty acid synthase (fas) as well as acetyl-CoA carboxylase (acc) among all treatments. Relative gene expression levels of 6-phosphogluconate dehydrogenase (6pgd) as well as glucose-6-phosphate dehydrogenase (g6pd) have been significantly upregulated in 0.40 and 2.50 g kg -1 ARA groups. Relative gene expression level of fatty acid binding protein 1 (fabp1) significantly increased in 4.60, 8.90, 12.50, and 15.70 g kg -1 ARA groups. However, the gene expression levels of fatty acid binding protein 4 (fabp4) as well as scavenger receptor class 2 (srb2) have not been influenced by dietary ARA levels. What is more, crabs fed diets with 4.60, 8.90, 12.50, and 15.70 g kg -1 ARA had a significantly higher expression level of carnitine palmitoyltransferase 1 (cpt1) than those fed diets with 0.40 and 2.50 g kg -1 ARA. In summary, optimum dietary ARA can promote growth, enhance antioxidant capacity, and improve health of mud crab juveniles. It also demonstrated that lipogenesis has been restrained with the increasing dietary ARA levels. These findings could provide theoretical guidance and reference for the lipid nutrition research as well as the development of the commercial diet in mud crab., Competing Interests: All authors declare no conflict of interest., (Copyright © 2022 Fang Fang et al.)

An 8-week feeding trial was carried out to assess the effect of dietary krill meal on growth performance and expression of genes related to TOR pathway and antioxidation of swimming crab ( Portunus trituberculatus ). Four experimental diets (45% crude protein and 9% crude lipid) were formulated to obtain different replacements of fish meal (FM) with krill meal (KM); FM was replaced with KM at 0% (KM0), 10% (KM10), 20% (KM20), and 30% (KM30); fluorine concentration in diets were analyzed to be 27.16, 94.06, 153.81, and 265.30 mg kg -1 , respectively. Each diet was randomly divided into 3 replicates; ten swimming crabs were stocked in each replicate (initial weight, 5.62 ± 0.19 g). The results indicated that crabs fed with the KM10 diet had the highest final weight, percent weight gain (PWG), and specific growth rate (SGR) among all treatments ( P < 0.05). Crabs fed with the KM0 diet had the lowest activities of total antioxidant capacity (T-AOC), total superoxide dismutase (SOD), glutathione (GSH), and hydroxyl radical scavenging activity and had the highest concentration of malondialdehyde (MDA) in the hemolymph and the hepatopancreas ( P < 0.05). In the hepatopancreas, the highest content of 20:5n-3 (EPA) and the lowest content of 22:6n-3 (DHA) were shown in crabs fed with the KM30 diet among all treatments ( P < 0.05). With the substitution level of FM with KM gradually increasing from 0% to 30%, the color of the hepatopancreas changed from pale white to red. Expression of tor , akt , s6k1 , and s6 in the hepatopancreas was significantly upregulated, while 4e-bp1 , eif4e1a , eif4e2 , and eif4e3 were downregulated with dietary replacement of FM with KM increasing from 0% to 30% ( P < 0.05). Crabs fed with the KM20 diet had notably higher expression of cat , gpx , cMnsod , and prx than those fed with the KM0 diet ( P < 0.05). Results demonstrated that 10% replacement of FM with KM can promote growth performance and antioxidant capacity and notably upregulate the mRNA levels of genes related to TOR pathway and antioxidant of swimming crab., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Chen Guo et al.)

The physiological processes involved in adaptation to osmotic pressure in euryhaline crustaceans are highly energy demanding, but the effects of dietary lipids (fat) on low salinity adaptations have not been well evaluated. In the present study, a total of 120 mud crabs ( Scylla paramamosain , BW = 17.87 ± 1.49 g) were fed control and high-fat (HF) diets, at both medium salinity (23‰) and low salinity (4‰) for 6 wk, and each treatment had 3 replicates with each replicate containing 10 crabs. The results indicated that a HF diet significantly mitigated the reduction in survival rate, percent weight gain and feed efficiency induced by low salinity ( P  < 0.05). Low salinity lowered lipogenesis and activated lipolysis resulting in lipid depletion in the hepatopancreas of mud crabs ( P  < 0.05). Thus, HF diets enhanced the process of lipolysis to supply more energy. In the gills, low salinity and the HF diet increased the levels of mitochondrial biogenesis markers, the activity of mitochondrial complexes, and the expression levels of genes related to energy metabolism ( P  < 0.05). Consequently, the positive effects of the HF diet on energy metabolism in mud crabs at low salinity promoted osmotic pressure regulation. Specifically, significantly higher haemolymph osmotic pressure and inorganic ion content, as well as higher osmotic pressure regulatory enzyme activity in gills, and gene and protein expression levels of NaK-ATPase were observed in crabs fed the HF diet at low salinity ( P  < 0.05). In summary, high dietary lipid levels improved energy provision to facilitate mitochondrial biogenesis, which increased ATP provision for osmotic pressure regulation of mud crabs. This study also illustrates the importance of dietary lipid nutrition supplementation for low salinity adaptations in mud crabs., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2023 The Authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd.)

In order to further explain the fillet texture improvement of Yellow River carp (Cyprinus carpio) fed with faba bean (Vicia faba L.), a three-month rearing trial was conducted to investigate fatty acid composition, antioxidant capacity, myofiber development, collagen deposition and transcriptome in white muscle of two farmed carp groups (One was fed only faba bean, the other was fed commercial diet). As a strong oxidant, faba bean changed fatty acids composition in white muscle, especially DHA and EPA, up-regulated the levels of reactive oxygen species (ROS) and down-regulated major antioxidant enzyme activities in the hepatopancreas and white muscle. Through the analysis of transcriptome and subsequent verification analysis, we speculated that the increase of ROS led to the decrease of myofiber diameter and collagen metabolism. This study provides a theoretical basis for further understanding the regulation of faba bean on fillet texture characteristic of Yellow River carp., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Carbohydrate is the cheapest source of energy among the three major nutrient groups, an appropriate amount of carbohydrates can reduce feed cost and improve growth performance, but carnivorous aquatic animals cannot effectively utilize carbohydrates. The objectives of the present study are aimed at exploring the effects of dietary corn starch levels on glucose loading capacity, insulin-mediated glycemic responses, and glucose homeostasis for Portunus trituberculatus . After two weeks of feeding trial, swimming crabs were starved and sampled at 0, 1, 2, 3, 4, 5, 6, 12, and 24 hours, respectively. The results indicated that crabs fed diet with 0% corn starch exhibited lower glucose concentration in hemolymph than those fed with the other diets, and glucose concentration in hemolymph remained low with the extension of sampling time. The glucose concentration in hemolymph of crabs fed with 6% and 12% corn starch diets reached the peak after 2 hours of feeding; however, the glucose concentration in hemolymph of crabs fed with 24% corn starch attained the highest value after 3 hours of feeding, and the hyperglycemia lasted for 3 hours and decreased rapidly after 6 hours of feeding. Enzyme activities in hemolymph related to glucose metabolism such as pyruvate kinase (PK), glucokinase (GK), and phosphoenolpyruvate carboxykinase (PEPCK) were significantly influenced by dietary corn starch levels and sampling time. Glycogen content in hepatopancreas of crabs fed with 6% and 12% corn starch first increased and then decreased; however, the glycogen content in hepatopancreas of crabs fed with 24% corn starch significantly increased with the prolongation of feeding time. In the 24% corn starch diet, insulin-like peptide (ILP) in hemolymph reached a peak after 1 hour of feeding and then significantly decreased, whereas crustacean hyperglycemia hormone (CHH) was not significantly influenced by dietary corn starch levels and sampling time. ATP content in hepatopancreas peaked at 1 h after feeding and then decreased significantly in different corn starch feeding groups, while the opposite trend was observed in NADH. The activities of mitochondrial respiratory chain complexes I, II, III, and V of crabs fed with different corn starch diets significantly increased first and then decreased. In addition, relative expressions of genes related to glycolysis, gluconeogenesis, glucose transport, glycogen synthesis, insulin signaling pathway, and energy metabolism were significantly affected by dietary corn starch levels and sampling time. In conclusion, the results of the present study reveal glucose metabolic responses were regulated by different corn starch levels at different time points and play an important role in clearing glucose through increased activity of insulin, glycolysis, and glycogenesis, along with gluconeogenesis suppression., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Xiangsheng Zhang et al.)

Cholesterol, as an indispensable nutrient, regulates molting and growth in crustacean. As crustaceans are unable to biosynthesize cholesterol de novo, it is central to understand how dietary cholesterol affects molting in crustaceans. An 8-week feeding trial was conducted to evaluate the effects of dietary cholesterol level (0.12%, 0.43%, 0.79%, 1.00%, 1.30% and 2.50%) on growth, cholesterol metabolism and expression of genes related to lipid and ecdysone metabolism in female swimming crabs ( Portunus trituberculatus ). A total of 192 crabs (1.41 ± 0.05 g) were randomly distributed into 192 aquaria. Each treatment had 4 replicates with each replicate containing 8 crabs. Crabs fed the 1.00% cholesterol diet showed best growth performance, and thus based on percent weight gain, the optimal dietary cholesterol requirement was calculated at 1.01%. Tissue cholesterol concentrations were positively correlated with dietary cholesterol level. The contents of functional fatty acids in hepatopancreas significantly increased as dietary cholesterol increased from 0.12% to 2.50% ( P  < 0.05). The expression levels of genes related to lipogenesis pathway, lipid catabolism and fatty acid oxidation were significantly down-regulated with increased dietary cholesterol level ( P  < 0.05). The highest expression levels of cholesterol transport genes, low-density lipoprotein receptor ( ldlr ) and low-density lipoprotein receptor-related protein 2 ( lrp2 ) occurred in crabs fed the 1.30% cholesterol diet. Moreover, hormones related to molting such as crustacean hyperglycemic hormone (CHH), methyl farnesoate (MF), molt-inhibiting hormone (MIH), and ecdysone in hemolymph were significantly influenced by dietary cholesterol level ( P  < 0.05). The highest expression levels of ecdysone receptor ( ecr ) and chitinase 1 ( chi1 ) in eyestalk and hepatopancreas were found in crabs fed the diet containing 1.00% cholesterol ( P  < 0.05). In conclusion, the optimal dietary level was beneficial to functional fatty acid accumulation, regulated lipid metabolism, promoted the ecdysone signalling pathway by improving the cholesterol transport, and improved the molting rate and growth of swimming crabs., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2022 Chinese Association of Animal Science and Veterinary Medicine. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd.)

Nonlinear process modeling is a primary task in intelligent manufacturing, aiming at extracting high-value features from massive process data for further process analysis like process monitoring. However, it is still a challenge to develop nonlinear process models with robust representation capability for diverse process faults. From the new perspective of the correlation between process variables, this paper develops a nonlinear process modeling algorithm to adaptively preserve the features of both global and local inter-variable structures, in order to fully exploit inter-variable features for enhancing the nonlinear representation of process operating conditions. Specifically, a unidimensional convolutional operation with a self-attention mechanism is proposed to simultaneously extract global and local inter-variable structures, wherein different attentions can be adaptively adjusted to these two structures for the final aggregation of them. Besides, cooperating with a two-dimensional dynamic data extension, the unidimensional convolutional operation can represent the overall temporal relationship between process samples. Through stacking a collection of these convolutional operations, a ResNet-style convolutional neural network then is constructed to extract high-order nonlinear features. Experiments on the Tennessee Eastman process validate the effectiveness of the proposed algorithm for two vital process monitoring problems-fault detection and fault identification., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 ISA. Published by Elsevier Ltd. All rights reserved.)

The quality of crustaceans' flesh has direct impact on consumers' purchase choices, with water environment and dietary nutrition being effective ways to regulate flesh quality. The aim of present study was to investigate the impacts of water salinity (low, 4 and medium, 23) and dietary lipid source (fish oil and soybean oil) on nutritional values, texture, taste and odor of flesh of mud crab. While water salinity had no significant influence on nutritional values of crab flesh, crabs fed soybean oil displayed significantly lower contents of amino acids and n-3 PUFAs in muscle. However, crabs reared at low salinity showed reduced flesh hardness, chewiness and gumminess likely related to altered myofiber structure, that impacted muscle texture. Furthermore, low salinity and dietary soybean oil weakened umami taste and aroma characteristics of crab flesh associated with decreased contents of free amino acids, flavor nucleotides, inorganic ions and odor active compounds in flesh., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

While tissue fatty acid compositions reflect that of the dietary lipid source, little information is available on how dietary oils modify lipid class and molecular species profiles in hepatopancreas of crustacean. Herein, an 8-week nutritional trial and untargeted lipidomic analysis were used to investigate the impacts of dietary n-3 PUFA lipid sources including fish oil, krill oil and linseed oil on the lipidomic characteristics of hepatopancreas of swimming crab (Portunus trituberculatus). Dietary krill oil significantly increased distribution of 20:5n-3 and 22:6n-3 at sn-2 in phosphatidylcholine and phosphatidylethanolamine compared to fish oil. Fish oil intake promoted the deposition of 20:5n-3 and 22:6n-3 at sn-1,2,3 in triglyceride compared to linseed oil, which significantly increased the specific accumulation of 18:3n-3 at sn-1,3 in triglyceride and sn-2 in phosphatidylcholine and phosphatidylethanolamine. The study revealed metabolic responses to different dietary n-3 PUFA in swimming crab, which provided novel insight into the lipid nutrition of crustacean., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

An 8-week feeding trial was conducted to evaluate the effects of dietary n-3 LC-PUFA levels on growth performance, tissue fatty acid profiles and relative expression of genes involved in the lipid metabolism of mud crab (Scylla paramamosain). Ten isonitrogenous diets were formulated to contain five n-3 LC-PUFA levels at 7 and 12 % dietary lipid levels. The highest weight gain and specific growth rate were observed in crabs fed the diets with 19·8 and 13·2 mg/g n-3 LC-PUFA at 7 and 12 % lipid, respectively. Moisture and lipid contents in hepatopancreas and muscle were significantly influenced by dietary n-3 LC-PUFA at the two lipid levels. The DHA, EPA, n-3 LC-PUFA contents and n-3:n-6 PUFA ratio in hepatopancreas and muscle significantly increased as dietary n-3 LC-PUFA levels increased at both lipid levels. The expression levels of -6 fatty acyl desaturase and acyl-CoA oxidase in hepatopancreas increased significantly, and expression levels of fatty acid synthase, carnitine palmitoyltransferase I and hormone-sensitive TAG lipase were down-regulated, with increased dietary n-3 LC-PUFA regardless of lipid level. Based on weight gain, n-3 LC-PUFA requirements of S. paramamosain were estimated to be 20·1 and 12·7 mg/g of diet at 7 and 12 % dietary lipid, respectively. Overall, dietary lipid level influenced lipid metabolism, and purified, high-lipid diets rich in palmitic acid reduced the n-3 LC-PUFA requirement of juvenile mud crab.

Salinity is one of the important factors affecting the physiological state of crustaceans in marine environments. Lipid plays major roles in energy supply and is main sources of essential fatty acids for membrane integrity, which is critical in adaptations to changes in salinity. Here we evaluated the effects of salinity (medium, 23 ppt and low, 4 ppt) and dietary lipid source (fish oil, FO and soybean oil, SO) on intestinal health of the marine crustacean mud crab Scylla paramamosain. The results indicated that low salinity and dietary SO (LSO group) significantly affected intestinal histomorphology, with a significant decrease of intestinal fold height and width as well as down-regulation of intestinal mRNA levels of tight junction genes compared to crab reared at medium salinity and fed FO diets (MFO group). Crabs reared at low salinity and fed SO showed an increased inflammatory response in intestine, which stimulated a physiological detoxification response together with apoptosis compared to crab in the MFO group. Low salinity and SO diets also could be responsible for multiply the pathogenic bacteria of Photobacterium and inhibit the beneficial bacteria of Firmicutes and Rhodobacteraceae in intestine, and act on a crucial impact on the development of intestinal microbial barrier disorders. The results of microbial function predictive analysis also support these inferences. The findings of the present study demonstrated that soybean oil as the main dietary lipid source could exacerbate the adverse effects of low salinity on intestinal health of mud crab, and provided evidence suggesting that dietary lipid source and fatty acid composition may play vital roles in intestinal health and the process of adaptation to environmental salinity in marine crustaceans., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Salinity has been demonstrated to influence the biosynthesis of long-chain (C 20-24 ) polyunsaturated fatty acids (LC-PUFAs) in teleost fish. Since LC-PUFAs are essential nutrients for vertebrates, it is central to understand how fish cope with an acute change in salinity associated with natural events. We herein report on the cloning and functional characterization of two elongation of very-long-chain fatty acid (Elovl)4 proteins, namely, Elovl4a and Elovl4b, and study the roles that these enzymes play in the biosynthesis of LC-PUFAs and very-long-chain (>C 24 ) polyunsaturated fatty acids (VLC-PUFAs) in marine teleost Pampus argenteus . The P. argenteus Elovl4 displayed all of the typical features of Elovl-like enzymes and have eyes and brain as major sites through which they exert their functions. Moreover, functional studies showed that the P. argenteus Elovl4 can effectively elongate C 18-22 substrates to C 36 VLC-PUFA. Because both P. argenteus Elovl4 are able to produce 24:5 n - 3 from shorter precursors, we tested whether the previously reported Δ6 Fads2 from P. argenteus was able to desaturate 24:5 n - 3 to 24:6 n - 3, a key step for docosahexaenoic acid (DHA) synthesis. Our results showed that P. argenteus can indeed bioconvert 24:5 n - 3 into 24:6 n - 3, suggesting that P. argenteus has the enzymatic capacity required for DHA biosynthesis through the coordinated action of both Elovl4 and Fads2. Furthermore, an acute salinity test indicated that low-salinity stress (12 ppt) upregulated genes involved in LC-PUFA biosynthesis, with 12 ppt salinity treatment showing the highest hepatic LC-PUFA content. Overall, our results unveiled that the newly characterized Elovl4 enzymes have indispensable functions in LC- and VLC-PUFA biosynthesis. Moreover, acute salinity change influenced the biosynthesis of LC-PUFA in P. argenteus . This study provided new insight into the biosynthesis of LC- and VLC-PUFAs in vertebrates and the physiological responses that teleosts have under acute salinity stress.

The aim of the present study was to investigate the effects of dietary Zn level on growth performance, Zn bioaccumulation, antioxidant capacity and innate immunity in juvenile mud crabs (Scylla paramamosain). Six semi-purified diets were formulated to contain dietary Zn levels of 44·5, 56·9, 68·5, 97·3, 155·6 or 254·7 mg/kg. Dietary Zn level significantly influenced percentage weight gain (PWG), with the highest observed in crabs fed the diet containing 97·3 mg/kg Zn. Tissue Zn concentrations significantly increased as dietary Zn levels increased from 44·5 to 254·7 mg/kg. Retention of Zn in hepatopancreas increased with dietary Zn levels up to 68·5 mg/kg and then significantly decreased. Moreover, inadequate dietary Zn (44·5 and 56·9 mg/kg) reduced antioxidation markers including total superoxide dismutase (SOD) and Cu/Zn SOD activities and total antioxidant level. Crabs fed the diet with 44·5 mg/kg Zn also showed significantly lower expression of genes involved in antioxidant status, such as Cu/Zn SOD, glutathione peroxidase, catalase and thioredoxin than those fed diets containing 68·5 and 97·3 mg/kg Zn. The highest activities of phenoloxidase and alkaline phosphatase were recorded in crabs fed the diets containing 68·5 and 97·3 mg/kg Zn. Expression levels of prophenoloxidase and toll-like receptor 2 were higher in crabs fed the 97·3 mg/kg Zn diet compared with crabs fed the other diets. Based on PWG alone, the optimal dietary Zn level was estimated to be 82·9 mg/kg, with 68·5 to 97·3 mg/kg recommended for maintaining optimal Zn bioaccumulation, oxidation resistance and innate immune response of juvenile mud crabs.

Copper is a widespread pollutant in marine environments, and marine animals can ingest large amounts of copper through the food chain. Here, an 8-week feeding trial was designed to investigate the effects of different dietary copper levels on coloration, copper bioaccumulation, stress response and oxidation resistance of juvenile mud crab Scylla paramamosain. The results indicated that crabs fed the diet with 162 mg/kg copper exhibited a dark-blue carapace and hemolymph. The accumulation of copper in tissues was positively correlated with the level of copper in feed. High/excess dietary copper (162 mg/kg) up-regulated the expression of stress response related genes, and reduced the expression/activities of anti-oxidation genes/enzymes. The activity of phenoloxidase decreased significantly when dietary copper level was 86-162 mg/kg, and the expression of hemocyanin was up-regulated in crab fed the diets with 28-162 mg/kg copper. Overall, the results of the present study indicated that high dietary copper led to parachrea in carapace and hemolymph of mud crab, and caused copper deposition abnormality in carapace and hepatopancreas. The data suggested that the toxic effects of dietary copper were concentration-dependent such that, excess dietary copper (162 mg/kg) had adverse impacts on oxidation resistance., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

An 8-week feeding trail was conducted in Acanthopagrus schlegelii with an initial body weight of 8.34 ± 0.01g. Three isonitrogenous diets were formulated, (1) Control: medium-fat diet (12%); (2) HFD: high-fat diet (18%); (3) HFD + FF: high-fat diet with fenofibrate (0.15%). Liver histological analysis revealed that, compared to HFD, vacuolar fat drops were smaller and fewer in fish fed fenofibrate. Expression of lipid catabolism regulator peroxisome proliferator-activated receptor alpha (pparα) was up-regulated by fenofibrate compared with HFD. In addition, fenofibrate significantly increased the expression level of silent information regulator 1 (sirt1). Meanwhile, the expression level of anti-inflammatory cytokine interleukin 10 (il-10) in intestine was up-regulated, while pro-inflammatory cytokine interleukin 1β (il-1β) in liver and intestine were down-regulated by dietary fenofibrate supplementation. Overall, the present study indicated that fenofibrate reduced fat deposition and attenuated inflammation response caused by HFD partly through a pathway involving regulation of pparα and sirt1., Competing Interests: Declaration of competing interest The authors have declared that no competing interests exist., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

An 8-week feeding trial was conducted to evaluate the effects of dietary xylanase supplementation on growth performance, digestive enzyme activity, intestinal morphology parameter, intestinal microbiome diversity, and carbohydrate metabolism for juvenile large yellow croaker (Larimichthys crocea). Four levels of xylanase were added to basal diets (0, 600, 1200, and 1800 U kg -1 ). The results indicated that fish fed the 1200 U kg -1 xylanase diet had higher weight gain than those fed the 0 and 600 U kg -1 xylanase diet. The highest intestinal folds and microvillous height were observed at fish fed the 1200 U kg -1 xylanase diet. High-throughput sequencing revealed that the majority of reads derived from the large yellow croaker digesta belonged to members of Proteobacteria followed by Chloroflex, Bacteroidetes, Spirochaetae, and Firmicute. Supplementation of xylanase in diets increased the relative abundance of Bacteroides and Gemmatimonadete. The higher hepatic glucokinase (GK) and glucose-6-phosphate dehydrogenase (G6PD) activities were observed in fish fed the xylanase supplementation diet. Accordingly, dietary xylanase supplementation upgraded the relative expressions of gk and g6pd genes in liver. In conclusion, optimum dietary xylanase supplementation (600-1200 U kg -1 ) could improve the growth performance, optimize the intestinal morphology structure and microbiota constitution, and enhance the ability of carbohydrate utilization of juvenile large yellow croaker.