Effects of supplementing coated methionine in a high plant-protein diet on growth, antioxidant capacity, digestive enzymes activity and expression of TOR signaling pathway associated genes in gibel carp, Carassius auratus gibelio.

This study explored the impacts of supplementation of different levels of coated methionine (Met) in a high-plant protein diet on growth, blood biochemistry, antioxidant capacity, digestive enzymes activity and expression of genes related to TOR signaling pathway in gibel carp (Carassius auratus gibeilo). A high-plant protein diet was formulated and used as a basal diet and supplemented with five different levels of coated Met at 0.15, 0.30, 0.45, 0.60 and 0.75%, corresponding to final analyzed Met levels of 0.34, 0.49, 0.64, 0.76, 0.92 and 1.06%. Three replicate groups of fish (initial mean weight, 11.37 ± 0.02 g) (20 fish per replicate) were fed the test diets over a 10-week feeding period. The results indicated that with the increase of coated Met level, the final weight, weight gain (WG) and specific growth rate initially boosted and then suppressed, peaking at 0.76% Met level (P< 0.05). Increasing dietary Met level led to significantly increased muscle crude protein content (P< 0.05) and reduced serum alanine aminotransferase activity (P< 0.05). Using appropriate dietary Met level led to reduced malondialdehyde concentration in hepatopancreas (P< 0.05), improved superoxide dismutase activity (P< 0.05), and enhanced intestinal amylase and protease activities (P< 0.05). The expression levels of genes associated with muscle protein synthesis such as insulin-like growth factor-1, protein kinase B, target of rapamycin and eukaryotic initiation factor 4E binding protein-1 mRNA were significantly regulated, peaking at Met level of 0.76% (P< 0.05). In conclusion, supplementing optimal level of coated Met improved on fish growth, antioxidant capacity, and the expression of TOR pathway related genes in muscle. The optimal dietary Met level was determined to be 0.71% of the diet based on quadratic regression analysis of WG.

Alpha-ketoglutaric acid mitigates the detrimental effects of soy antigenic protein on the intestinal health and growth performance of Mirror carp Cyprinus carpio.

The study investigated the alleviated effects of Alpha-ketoglutaric acid (AKG) on the intestinal health of mirror carp (Cyprinus carpio Songpu) caused by soy antigenic protein. The diets were formulated from fishmeal (CON), 50% soybean meal (SBM), the mixture of glycinin and ß-conglycinin (11 + 7S) and adding 1% AKG in the 11 + 7S (AKG). Carp (~ 4 g) in triplicate (30 fish per tank) was fed to apparent satiation thrice a day for six weeks. Compared with CON, SBM treatment resulted in significantly poor growth performance (P < 0.05), whereas 11 + 7S and AKG treatments were not significantly different from CON (P > 0.05). Gene expression of tumor necrosis factor (TNF-α) and interleukin-1 ß (IL-1ß) in proximal intestines (PI) and distal intestines (DI) were increased (P < 0.05), and transforming growth factor (TGF-ß) in PI and middle intestines (MI) was decreased (P < 0.05) in both SBM and 11 + 7S. The caspase-3 in DI increased in SBM (P < 0.05) and the caspase-3 and caspase-9 in DI increased in 11 + 7S (P < 0.05); conversely, TGF-ß in PI and MI was increased, TNF-α and IL-1ß in the MI, caspase-3, and caspase-9 in DI was decreased in AKG (P < 0.05). The TOR (target of rapamycin) in PI and MI, ACC in PI, MI and DI was decreased in SBM (P < 0.05), the AMPK in the PI and DI, TOR in PI, MI and DI, ACC in PI and DI, 4E-BP in DI was reduced in 11 + 7S (P < 0.05). AMPK in the PI and DI, ACC in the PI and MI, TOR in PI, MI, and DI, 4E-BP in PI and DI was recovered by AKG supplementation (P < 0.05). Lipids and lipid-like metabolism, organic acids and derivatives metabolism increased in AKG dietary treatment. In conclusion, AKG reduces the expression of intestinal inflammation and apoptosis pathway and changes glycerophospholipid metabolism and sphingolipid metabolism in the intestine of fish.

Combined effects of dietary carbohydrate levels and ammonia stress on growth, antioxidant capacity and glucose metabolism in juvenile oriental river prawn (Macrobrachium nipponense).

Ammonia is a common environmental stress factor that constrains aquaculture industry development. This study evaluated the effect of carbohydrate levels and ammonia stress in oriental river prawn (Macrobrachium nipponense). The experiment had six treatments containing two water ammonia levels (0 and 5 mg/L) and three dietary carbohydrate levels (low carbohydrate diet (LCD, 10%), medium carbohydrate diet [MCD, 20%], and high carbohydrate diet [HCD, 30%]), and lasted six weeks. The results showed that the prawns fed on MCD had higher weight gain than those fed on LCD and HCD during ammonia stress. Moreover, the prawns fed on MCD had significantly lower acid phosphatase and alkaline phosphatase activities during ammonia stress. Feeding the prawns on the MCD increased B cells in the hepatopancreas during ammonia stress. Interestingly, the prawns fed on MCD had significantly lower superoxide dismutase activity compared to LCD and HCD during ammonia stress. Moreover, the prawns fed on MCD had significantly lower pyruvate kinase activity and pyruvate and lactic acid contents, while those fed on LCD had significantly higher succinic dehydrogenase, 6-phosphogluconic dehydrogenase, and phosphoenol pyruvate carboxykinase activities during ammonia stress. The prawns fed on the MCD increased significantly glutaminase activity and decreased the ammonia content in the serum during ammonia exposure. In addition, feeding the prawns on MCD decreased significantly the expression of apoptosis and inflammation-related genes. Taken together, the MCD supplied energy required to counteract ammonia stress, which increased growth, improved antioxidant capacity, facilitated ammonia excretion, and alleviated inflammation and apoptosis of the oriental river prawn.

Effects of Replacing Fish Meal with Enzymatic Cottonseed Protein on the Growth Performance, Immunity, Antioxidation, and Intestinal Health of Chinese Soft-Shelled Turtle (Pelodiscus sinensis).

The dietary effects of replacing fish meal with enzymatic cottonseed protein (ECP) on the growth performance, immunity, antioxidant, and intestinal health of Chinese soft-shelled turtles have not been explored. An eight-week feeding trial was conducted with a quadruplicated group of turtles (3.44 ± 0.01 g) that were randomly assigned to 16 cages (0.6 m × 0.6 m × 0.6 m) with 30 turtles that were stocked in each cage. Four dietary groups were fed with diets supplemented with 0, 2%, 4%, and 6% (ECP0 group (control group), ECP2 group, ECP4 group, ECP6 group) of enzymatic cottonseed protein replacing fishmeal. The present study illustrated that the final weight and WG in the ECP2 and ECP4 groups were significantly increased (P < 0.05) compared with the control group. The ECP2, ECP4, and ECP6 groups significantly reduced the feed coefficient (P < 0.05) and significantly increased the SGR (P < 0.05). The serum TP and ALB of the ECP4 group were significantly increased (P < 0.05). The ECP2, ECP4, and ECP6 groups significantly increased the activity of intestinal pepsin (P < 0.05), and the activity of intestinal lipase of the EPC4 group was significantly increased (P < 0.05). The intestinal villus height of the EPC4 group and EPC6 group, the villus width of the EPC2 group and EPC4 group, and the intestinal muscle thickness of the EPC4 group were significantly increased (P < 0.05). At the same time, replacing fishmeal with enzymatic cottonseed protein also affected the intestinal inflammation-related genes compared with the control group. Besides that, the expression of the IL-10 gene in the experimental group was significantly upregulated (P < 0.05). Nevertheless, the expression of TNF-α and IL-8 genes in the ECP2 group and TNF-α and IL-1ß genes in the ECP4 group was significantly downregulated (P < 0.05). In summary, replacing fish meal with enzymatic cottonseed protein positively affects the growth, immunity, and intestinal health of Chinese soft-shelled turtles. The appropriate proportion of enzymatic cottonseed protein to replace fish meal in turtle feed is 4%.

Dietary α-ketoglutarate alleviates glycinin and ß-conglycinin induced damage in the intestine of mirror carp (Cyprinus carpio).

This study investigated the glycinin and ß-conglycinin induced intestinal damage and α-ketoglutarate alleviating the damage of glycinin and ß-conglycinin in intestine. Carp were randomly divided into six dietary groups: containing fish meal (FM) as the protein source, soybean meal (SM), glycinin (FMG), ß-conglycinin (FMc), glycinin+1.0% α-ketoglutarate (AKG) (FMGA), ß-conglycinin+1.0% AKG (FMcA). The intestines were collected on 7th, and the hepatopancreas and intestines were collected on 56th. Fish treated with SM and FMc displayed reduced weight gain, specific growth rate, and protein efficiency. On 56th day, Fish fed on SM, FMG and FMc presented lower superoxide dismutase (SOD) activities. FMGA and FMcA had higher SOD activity than those fed on the FMG and FMc, respectively. In intestine, fish fed on the SM diets collected on 7th presented upregulated the expression of transforming growth factor beta (TGFß1), AMP-activated protein kinase beta (AMPKß), AMPKγ, and acetyl-CoA carboxylase (ACC). Fish fed FMG presented upregulated expression of tumor necrosis factor alpha (TNF-α), caspase9, and AMPKγ, while downregulated the expression of claudin7 and AMPKα. FMc group presented upregulated expression of TGFß1, caspase3, caspase8, and ACC. Fish fed FMGA showed upregulated expression of TGFß1, claudin3c, claudin7, while downregulating the expression of TNF-α and AMPKγ when compared to fish fed FMG diet. FMcA upregulated the expression of TGFß1, claudin3c than fed on the FMc. In intestine, the villus height and mucosal thickness of the proximal intestine (PI) and the distal intestine (DI) were decreased and crypt depth of the PI and mid intestine (MI) were increased in SM, FMG and FMc. In addition, fish fed on SM, FMG and FMc presented lower citrate synthase (CS), isocitrate dehydrogenase (ICD), α-ketoglutarate dehydrogenase complex (α-KGDHC) Na+/K+-ATPase activity in DI. FMGA had higher CS, ICD, α-KGDHC, and Na+/K+-ATPase activity in PI and MI than those fed on the FMG. FMcA had higher Na+/K+-ATPase activity in MI. In conclusion, dietary soybean meal destroys the intestine's health, the adverse effects are related to the presence of ß-conglycinin and glycinin, especially glycinin. AKG may regulate intestinal energy via tricarboxylic acid cycle, thereby alleviating the damage intestinal morphology caused by the dietary soybean antigen proteins.

Effects of the Replacement of Dietary Fish Meal with Poultry By-Product Meal on Growth and Intestinal Health of Chinese Soft-Shelled Turtle (Pelodiscus sinensis).

To investigate the effect of poultry by-product meal (PBM) replacing fish meal on the growth and intestinal health of Chinese soft-shelled turtle (Pelodiscus sinensis). Four experimental diets were prepared. Fish meal was replaced by 0 (control group, PBM0), 5% (PBM5), 10% (PBM10), and 15% (PBM15) PBM. Compared to the control group, final body weight, weight gain, and specific growth rate were significantly increased, while feed conversion rate decreased significantly in the PBM10 group (p < 0.05). The PBM15 group significantly increased the moisture content and significantly decreased the ash content of the turtles (p < 0.05). The PBM5 and PBM15 groups significantly decreased the whole-body crude lipid (p < 0.05). The serum glucose content increased significantly in the PBM10 group (p < 0.05). The liver malonaldehyde content significantly decreased in the PBM5 group and in the PBM10 group (p < 0.05). Liver glutamic-oxalacetic transaminase and intestinal pepsin activity were increased significantly in the PBM15 group (p < 0.05). The expression of the intestinal interleukin 10 (IL-10) gene was significantly down-regulated in the PBM10 group and the PBM15 group (p < 0.05), the expression of the intestinal interferon-γ (IFN-γ), interleukin-8 (IL-8), and liver toll-like receptor 4 (TLR4) and toll-like receptor 5 (TLR5) genes were significantly up-regulated in the PBM5 group (p < 0.05). In summary, poultry by-product meal can be used as a protein source to replace fish meal in turtle feed. Based on quadratic regression analysis, the optimal replacement ratio is 7.39%.

Nuclease treatment enhanced the ameliorative effect of yeast culture on epidermal mucus, hepatic lipid metabolism, inflammation response and gut microbiota in high-fat diet-fed zebrafish.

As a functional feed additive, yeast cultures are rich in nucleotides, and adding extra nuclease can significantly increase the content of nucleotides in yeast culture. In this experiment, the effects on growth, epidermal mucus, liver and intestinal health of zebrafish were evaluated by supplementing the yeast culture or nuclease-treated yeast culture with a high-fat diet (HFD). One-month-old zebrafish were fed four diets: normal diet (NORM), HFD, yeast culture diet (YC), and nuclease-treated yeast culture diet (YC (N)) for three weeks. Results showed that the complement 4 activity of the epidermal mucus in YC (N) group was significantly higher than those in HFD and YC groups (P < 0.05). The YC and YC (N) significantly reduced the content of hepatic triglyceride caused by HFD (P < 0.05). Moreover, compared with the YC group, the YC (N) significantly increased the expression of lipolysis genes, such as PPARα, PGC1α, ACOX3 (P < 0.05). Compared with the YC group, the YC (N) group significantly increased the expression of liver pro-inflammatory factors TNFα, IL-6, IL-1ß and anti-inflammatory factors TGFß, IL-10 (P < 0.05). The diet YC and YC (N) significantly improved the height of the intestinal villus (P < 0.05). Compared with the HFD group, the YC (N) group significantly increased the expression of intestinal pro-inflammatory factors TNFα, IL-6 and anti-inflammatory factors TGFß, IL-10 (P < 0.05). The YC (N) group significantly decreased the abundance of intestinal Proteobacteria and Acinetobacter, and increased the abundance of intestinal Actinobacteria, Mycobacterium and Rhodobacter (P < 0.05). In conclusion, compared with the supplement of yeast culture, nuclease treated yeast culture can further alleviate the adverse effects of HFD on liver and intestinal health, and be used as feed additives for the nutritional and immune regulation of fish.

Low Protein Diets Supplemented With Alpha-Ketoglutarate Enhance the Growth Performance, Immune Response, and Intestinal Health in Common Carp (Cyprinus carpio).

To investigate the effects of alpha-ketoglutarate (AKG) supplementation in a low protein (LP) diet on the growth performance, immune response, and intestinal health of common carp (Cyprinus carpio), 600 carp were randomly divided into five dietary groups: a normal protein (NP) diet containing 32% crude protein, an LP diet formulated with 28% crude protein, and LP with AKG at 0.4%, 0.8%, and 1.2% (dry matter). After an 8-week trial period, the results demonstrated that an LP diet led to a decrease in performance, immune response, and intestinal barrier function. Compared with the LP group, the final body weight and weight gain rate in the LP+0.4% AKG group were significantly higher, the feed conversion ratio was significantly decreased with the addition of 0.4% and 0.8% AKG. The supplementation with 0.4% and 0.8% AKG markedly increased the activities of T-SOD and GSH-Px, as well as the expression levels of GPX1a and GPX1b relative to the LP group, whereas the MDA content was significantly decreased in the LP+0.4% AKG group. In addition, the expression levels of tight junctions including claudin-3, claudin-7, ZO-1, and MLCK were significantly up-regulated in the LP+0.4% AKG group, and the relative expression levels of the pro-inflammatory factors IL-1ß and IL-6α were significantly lower with the addition of 0.4%, 0.8%, and 1.2% AKG. Moreover, the abundance of Proteobacteria in the LP+0.4% AKG group was lower than that in the LP group, and the abundance of Firmicutes and Fusobacteria was higher at the phylum level. The abundance of Citrobacter in the LP+0.4% AKG group was decreased compared to the LP group, while the abundance of Aeromonas was increased at the genus level. In short, the effects of AKG on the intestinal health of the common carp were systematically and comprehensively evaluated from the perspectives of intestinal physical barrier, chemical barrier, biological barrier, and immune barrier. We found that an LP diet supplemented with 0.4% AKG was beneficial to the growth performance and intestinal health of common carp.

Assessment of chicken intestinal hydrolysates as a new protein source to replace fishmeal on the growth performance, antioxidant capacity and intestinal health of common carp (Cyprinus carpio).

Promoting circular economy by transforming food residues into alternative high-value protein sources for aquaculture feed is a new way to develop alternative raw materials for fishmeal. This study systematically evaluated the effects of chicken intestinal hydrolysates (CIH) on the intestinal immune health of common carp through growth performance, antioxidant capacity, and intestinal immunity analysis in order to replace fishmeal. Five iso-nitrogenous and iso-lipidic experimental feeds were formulated to replace 0% (CIH-0), 25% (CIH-25), 50% (CIH-50), 75% (CIH-75) and 100% (CIH-100) of the fishmeal with CIH. Each experimental diet was fed to triplicate groups of 30 carp for 8 weeks. The results revealed that no significant differences in the final body weight, weight gain rate, feed coefficient radio, feed intake and protein efficiency ratio were found among the CIH-0, CIH-25, and CIH-50 groups, while the final body weight and weight gain rate in the CIH-75 and CIH-100 groups were significantly decreased and the feed coefficient radio was significantly increased. The aspartate aminotransferase of all CIH groups were significantly decrease, and the total protein, albumin did not differ among the CIH-0, CIH-25, CIH-50, and CIH-75 groups. The trypsin content was significantly increased in the CIH-75 and CIH-100 groups. No significant differences in the antioxidant index (catalase, glutathione peroxidase and malonaldehyde) were found among all CIH groups compared with the CIH-0 group. The expression levels of pro-inflammatory cytokines IL-1ß and TNF-α were significantly down-regulated in the CIH-50 group and anti-inflammatory cytokines IL-10 and TGF-ß2 were significantly up-regulated in the CIH-50 and CIH-75 groups. No significant differences in the expression levels of claudin-1, claudin-7 and claudin-11 were observed between the CIH-0 and CIH-50 groups, while the expression levels of ZO-1, occludin and MLCK were significantly up-regulated in the CIH-50 group compared with the CIH-0 group. The expression level of claudin-1 was down-regulated in the CIH-75 and CIH-100 groups. Hence, the study demonstrated the potential of CIH as a novel protein source for replacing fishmeal, and replacing 50% of fishmeal with CIH did not significantly influence the growth performance, immune responses, and intestinal barrier of common carp (Cyprinus carpio).

Effect of phosphorus on growth performance, intestinal tight junctions, Nrf2 signaling pathway and immune response of juvenile mirror carp (Cyprinus carpio) fed different α-ketoglutarate levels.

Previous research has shown that dietary α-ketoglutarate (AKG) supplementation can promote growth performance, phosphorus metabolism, and skeletal development of juvenile mirror carp (Cyprinus carpio) fed low phosphorous diets. In the current study, we tested the hypothesis that 1% AKG dietary supplementation reduces the dietary phosphorus requirements of juvenile mirror carp. A total of 12 experimental isoproteic and isolipidic diets containing available phosphorus levels of 0.21%, 0.38%, 0.55%, 0.72%, 0.89%, and 1.07% dry matter with either 0 or 1% AKG supplementation were used in the study. A total of 1080 juvenile fish of similar initial weight (0.90 ± 0.03 g) were selected and randomly assigned to 36 tanks. There were three replicates for each experimental group, with a density of 30 fish per tank. Fish were fed to satiation for 8 weeks. The results indicated that fish fed the diet supplemented with 1% AKG showed a significant increase in final body weight (FBW), weight gain rate (WGR), feed intake (FI) and phosphorus intake (PI) compared to the diet without AKG (P < 0.05). FBW and WGR increased significantly with increasing available phosphorus levels from 0.21% to 0.89% (P < 0.05). The mRNA expression of ZO-1, claudin 11, and occludin was significantly increased by dietary AKG and phosphorus (P < 0.05). The mRNA expression of Nrf2, GPx1a, and CAT in the Nrf2 signaling pathway was significantly increased by dietary AKG and phosphorus (P < 0.05). The expression levels of IL-10 and TGF-ß2 were significantly increased by dietary AKG and phosphorus, but the expression levels of IL-1ß, IL-8, IL-10, TNF-a and NF-κB were significantly decreased with dietary AKG and phosphorus supplementation (P < 0.05). Based on second-order polynomial regression analysis of WGR against dietary phosphorus levels, the optimal dietary phosphorus level was found to be 0.79% of dry feed for juvenile mirror carp fed a diet with 1% AKG supplementation and 0.93% of dry feed without AKG supplementation. This study confirmed that AKG supplementation can reduce the phosphorus requirements of juvenile mirror carp by promoting growth performance, intestinal tight junctions, Nrf2 signaling pathways and immune response.

Evaluation of Alpha-Ketoglutarate Supplementation on the Improvement of Intestinal Antioxidant Capacity and Immune Response in Songpu Mirror Carp (Cyprinus carpio) After Infection With Aeromonas hydrophila.

As an intermediate substance of the tricarboxylic acid cycle and a precursor substance of glutamic acid synthesis, the effect of alpha-ketoglutarate on growth and protein synthesis has been extensively studied. However, its prevention and treatment of pathogenic bacteria and its mechanism have not yet been noticed. To evaluate the effects of alpha-ketoglutarate on intestinal antioxidant capacity and immune response of Songpu mirror carp, a total of 360 fish with an average initial weight of 6.54 ± 0.08 g were fed diets containing alpha-ketoglutarate with 1% for 8 weeks. At the end of the feeding trial, the fish were challenged with Aeromonas hydrophila for 2 weeks. The results indicated that alpha-ketoglutarate supplementation significantly increased the survival rate of carp after infection with Aeromonas hydrophila (P < 0.05), and the contents of immune digestion enzymes including lysozyme, alkaline phosphatase and the concentration of complement C4 were markedly enhanced after alpha-ketoglutarate supplementation (P < 0.05). Also, appropriate alpha-ketoglutarate increased the activities of total antioxidant capacity and catalase and prevented the up-regulation in the mRNA expression levels of pro-inflammatory cytokines including tumor necrosis factor-α, interleukin-1ß, interleukin-6, and interleukin-8 (P < 0.05). Furthermore, the mRNA expression levels of toll-like receptor 4 (TLR4), and nuclear factor kappa-B (NF-κB) were strikingly increased after infection with Aeromonas hydrophila (P < 0.05), while the TLR4 was strikingly decreased with alpha-ketoglutarate supplementation (P < 0.05). Moreover, the mRNA expression levels of tight junctions including claudin-1, claudin-3, claudin-7, claudin-11 and myosin light chain kinases (MLCK) were upregulated after alpha-ketoglutarate supplementation (P < 0.05). In summary, the appropriate alpha-ketoglutarate supplementation could increase survival rate, strengthen the intestinal enzyme immunosuppressive activities, antioxidant capacities and alleviate the intestinal inflammation, thereby promoting the intestinal immune responses and barrier functions of Songpu mirror carp via activating TLR4/MyD88/NF-κB and MLCK signaling pathways after infection with Aeromonas hydrophila.

Effect of Bacillus megaterium-Coated Diets on the Growth, Digestive Enzyme Activity, and Intestinal Microbial Diversity of Songpu Mirror Carp Cyprinus specularis Songpu.

The present study was conducted to evaluate the effect of a Bacillus megaterium-coated diet on growth performance, digestive enzymes, and intestinal microbial diversity in Songpu mirror carp (Cyprinus specularis Songpu). The fish were manually fed two diets (a control diet and a B. megaterium-coated diet) three times daily until apparent satiation for 56 days. Compared with the control group, supplementation with the B. megaterium-coated diet enhanced the fish growth and significantly reduced the feed conversion ratio (P < 0.05). The activities of foregut amylase and lipase in the treatment group were significantly higher than those in the control group (P < 0.05). The activities of foregut, midgut, and hindgut proteases in the treatment group were all higher than those in the control group (P > 0.05). The results of sequencing the 16S rDNA genes of the microbiota through high-throughput sequencing showed that the diversity and abundance of the intestinal microflora increased along with Songpu mirror carp growth. The Songpu mirror carp fed a diet coated with B. megaterium displayed increased proportions of intestinal Bacillus and Lactococcus at the genus level, and both were significantly higher than those of the control group (P < 0.05). These results therefore suggest that dietary B. megaterium application can improve the growth and digestive enzyme activity of Songpu mirror carp and enrich the beneficial genus composition of its main intestinal microflora.

Effect of Yucca schidigera extract on the growth performance, intestinal antioxidant status, immune response, and tight junctions of mirror carp (Cyprinus carpio).

The current study was designed to evaluate the effect of Yucca schidigera extract (YSE) on the growth performance, intestinal antioxidant status, immune response, and tight junctions of mirror carp (Cyprinus carpio). A total of 450 mirror carp (45.21 ± 0.43 g) were fed diets supplemented with 0, 200, or 400 mg/kg YSE for 8 weeks. Compared with the control (0 mg/kg), the final body weight and weight gain rate were significantly higher in the 400 mg/kg YSE group (P < 0.05), and the serum ammonia concentration was significantly lower in both YSE groups (P < 0.05). Additionally, the total antioxidant capacity was significantly higher in the 400 mg/kg YSE group (P < 0.05), and the malondialdehyde content was significantly lower in both YSE groups (P < 0.05). Complement 3 and 4 contents were significantly higher in the 400 mg/kg YSE group (P < 0.05), and lysozyme was significantly higher in both YSE groups compared to the control group (P < 0.05). The relative mRNA levels of copper zinc superoxide dismutase, catalase, glutathione peroxidase1a, and nuclear factor erythroid 2-related factor 2 as well as transforming growth factor ß were significantly higher in both YSE supplemented groups compared to the control (P < 0.05), whereas the relative mRNA level of Kelch-like ECH-associated protein 1 was significantly lower in both YSE supplemented groups (P < 0.05). The relative mRNA levels of interleukin 1ß and interleukin 6 were significantly lower in the 400 mg/kg YSE supplemented group compared to the control (P < 0.05). Additionally, both YSE levels decreased the relative mRNA expression of tumour necrosis factor-α (P < 0.05). The relative mRNA levels of ZO-1 and claudin 11 were significantly higher in both YSE supplemented groups (P < 0.05), and the relative mRNA level of occludin was significantly higher in the 200 mg/kg YSE group than the control and 400 mg/kg YSE groups (P < 0.05). In conclusion, dietary supplementation with 400 mg/kg YSE improved the growth, intestinal antioxidant status, immune response, and tight junctions of mirror carp.

Optimal dietary curcumin improved growth performance, and modulated innate immunity, antioxidant capacity and related genes expression of NF-κB and Nrf2 signaling pathways in grass carp (Ctenopharyngodon idella) after infection with Aeromonas hydrophila.

This study investigated the effects of dietary curcumin on growth performance, non-specific immunity, antioxidant capacity and related genes expression of NF-κB and Nrf2 signaling pathways in grass carp (Ctenopharyngodon idella). A total of 525 juvenile grass carps with mean initial body weight of (5.30 ± 0.10) g were randomly distributed into five groups with three replicates each, fed five diets containing graded levels of curcumin (0, 196.11, 393.67, 591.46 and 788.52 mg/kg diet) for 60 days. After feeding trial, fifteen fish per tank were challenged with Aeromonas hydrophila and the mortalities were recorded for 7 days. The results showed that optimal dietary curcumin (393.67 mg/kg diet) improved the weight gain (WG) and specific growth rate (SGR) of juvenile grass carp, reduced feed conversion ratio (FCR) and the mortalities after challenge (P < 0.05). Moreover, optimal dietary curcumin increased the activities of lysozyme (LYZ) and acid phosphatase (ACP), and complement 3 (C3) and C4 levels, decreased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum of grass carp after injection with A. hydrophila (P < 0.05). Meanwhile, optimal dietary curcumin up-regulated the mRNA levels of LYZ, C3 and antimicrobial peptides [hepcidin, liver-expressed antimicrobial peptide-2 (LEAP-2), ß-defensin], and anti-inflammatory cytokines of interleukin-10 (IL-10) and transforming growth factor ß1 (TGF-ß1), and inhibitor of κBα (IκBα), whereas down-regulated pro-inflammatory cytokines of tumor necrosis factor-α (TNF-α), IL-1ß, IL-6 and IL-8, and nuclear factor kappa B p65 (NF-κB p65), IκB kinases (IKKα, IKKß and IKKγ) mRNA levels in the liver and blood of grass carp after injection with A. hydrophila (P < 0.05). In addition, optimal dietary curcumin increased the reduced glutathione (GSH) content and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR), reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels in the liver of grass carp after injection with A. hydrophila (P < 0.05). Meanwhile, optimal dietary curcumin up-regulated the mRNA levels of these antioxidant enzymes and nuclear factor erythroid 2-related factor 2 (Nrf2), whereas down-regulated Kelch-like ECH-associated protein (Keap) 1a and Keap 1b mRNA levels (P < 0.05) in the liver and blood of grass carp after injection with A. hydrophila. Thus, optimal dietary curcumin supplementation could promote growth of juvenile grass carp, reduce FCR, and enhance disease resistance, innate immunity and antioxidant capacity of fish, attenuating inflammatory response. However, dietary excessive curcumin had negative effect on fish. Based on second-order regression analysis between dietary curcumin contents and weight gain, the optimum requirement of dietary curcumin in juvenile grass carp was determined to be 438.20 mg/kg diet.

Effect of N-carbamoylglutamate supplementation on the growth performance, antioxidant status and immune response of mirror carp (Cyprinus carpio) fed an arginine-deficient diet.

The present study was conducted to determine the effect of dietary N-carbamoylglutamate (NCG) supplementation on the growth performance, antioxidant capability and immune responses of mirror carp (Cyprinus carpio) fed an arginine (Arg)-deficient diet. A total of 630 mirror carp (41.65 ±â€¯0.14 g) were fed diets (Arg 1.24% of the diet) that were supplemented with 0.50% Arg (control diet) or graded levels of NCG at 0 (Arg deficiency diet), 0.04%, 0.08%, 0.12%, 0.16% and 0.20% for 8 weeks. The results showed that, compared with the control diet, the Arg-deficient diet supplementation with 0 NCG (1) decreased the final body weight (FWB), the weight gain rate (WGR) or the protein efficiency ratio (PER) and increased the feed conversion ratio (FCR); (2) decreased the concentration of Arg and nitric oxide (NO) and the activity of total nitric oxide synthetase (T-NOS) in the plasma; (3) decreased the activities of superoxide dismutase (SOD) in the proximal intestine (PI), catalase (CAT) in the PI and distal intestine (DI), and glutathione peroxidase (GPx) in PI and mid-intestine (MI) and increased the concentration of malondialdehyde (MDA) in the PI, MI and DI; and (4) decreased the activity of lysozyme in the plasma, increased the relative mRNA expression of tumor necrosis factor-α (TNF-α), interleukin1ß (IL-1ß) and interleukin 8 (IL-8) in the PI, MI and DI, and decreased the relative mRNA expression of interleukin 10 (IL-10) in the PI and MI, and transforming growth factor ß2 (TGF-ß2) in the PI, MI and DI. Compared with the Arg deficient-diet supplementation with 0 NCG, (1) 0.12% or 0.16% NCG increased the FBW, WGR and PER, and 0.16% NCG increased the FCR; (2) 0.08%-0.20% NCG increased the concentration of Arg, NO and the activity of T-NOS; (3) 0.08% NCG increased the activities of SOD in the PI and MI, and 0.12% NCG increased activities of CAT and GPx in the PI, MI and DI; and (4) 0.04%-0.20% NCG increased the activity of lysozyme, 0.04%-0.20% NCG decreased the relative mRNA expression of TNF-α, IL-1ß and IL-8 in the PI and MI, and 0.04%-0.20% NCG increased the relative mRNA expression of IL-10 and TGF-ß2 in the PI and MI. The present results indicated that dietary 0.12% or 0.16% NCG improved the growth performance, feed utilization, intestinal antioxidant capacity and immune response of mirror carp fed an Arg-deficient diet.

Respiratory response of grass carp Ctenopharyngodon idellus to dissolved oxygen changes at three acclimation temperatures.

Respiratory parameters of grass carp were studied during dissolved oxygen (DO) changes from normal DO to hypoxia, then return to normal DO at 15, 25, and 30 °C acclimation, respectively. The results showed that with increases of acclimation temperature at normoxia the respiratory frequency (fR), oxygen consumption rate (VO2), respiratory stroke volume (VS.R), gill ventilation (VG), and VG/VO2 of grass carp increased significantly, but the oxygen extraction efficiency (EO2) of fish decreased significantly (P < 0.05). With declines of DO levels, the fR, VS.R, VG, and VG/VO2 of fish increased significantly at different acclimation temperatures (P < 0.05). A slight increase was found in VO2, and the EO2 of fish remained almost constant above DO levels of 3.09, 2.91, and 2.54 mg l-1 at 15, 25, and 30 °C, while the VO2 and EO2 began to decrease significantly with further reductions in DO levels (P < 0.05). After 0.5 h of recovery to normoxia from hypoxia at three acclimation, the fR, VS.R, VG, and VG/VO2 of the fish decreased sharply; meanwhile, the VO2 and EO2 increased sharply (P < 0.05). The respiratory parameters of fish gradually approached initial values with prolonged recovery time to normoxia, and reached their initial values in 2.5 h at 25 and 30 °C acclimation. The critical oxygen concentrations (Cc) of fish for VO2 were 2.42 mg l-1 at 15 °C, 2.02 mg l-1 at 25 °C, and 1.84 mg l-1 at 30 °C, respectively. The results suggest that grass carp are highly adapted to varied DO and short-term hypoxia environments.

Effects of alpha-ketoglutarate (AKG) on growth performance and non-specific immunity of juvenile red drum fed diets with low or adequate phosphorus levels.

This study evaluated the effects of α-ketoglutarate (AKG) on the growth performance, body, and skeletal composition, as well as non-specific immunity of juvenile red drum (Sciaenops ocellatus) fed adequate or low-phosphorus diets. A 2 × 2 factorial design was arranged with two levels of total phosphorus (1.0 or 1.6%) and two levels (0 or 1%) of AKG. Each diet was fed to red drum in four replicate 110-L aquaria (15 fish/aquarium) twice daily for 7 weeks. The results indicated that the low-P diets significantly (P < 0.05) decreased weight gain, feed efficiency, and condition factor of red drum, but increased moisture content and decreased ash content of whole body, as well as decreased ash content of skeletal tissue and decreased plasma lysozyme. Supplementation of AKG significantly improved the fillet yield and plasma lysozyme and tended to improve neutrophil oxidative radical production (P = 0.097). Based on these results, phosphorus level had the greatest effect on growth performance and non-specific immunity of red drum, while AKG supplementation had limited positive effects on immunological responses and fillet yield of juvenile red drum.

Effects of the supplementation of vitamin D3 on the growth and vitamin D metabolites in juvenile Siberian sturgeon (Acipenser baerii).

The objective of this study was to evaluate the effects of the supplementation of vitamin D3 on the growth, vitamin D metabolites, and osteocalcin secretion in juvenile Siberian sturgeon (Acipenser baerii). A 90-day growth trial was conducted with juvenile Siberian sturgeon (initial body weight 3.47 ± 0.14 g) fed seven isonitrogenous and isoenergetic practical diets (45% CP and 13% lipid) containing 60 (basal diet), 240, 450, 880, 1670, 3300, or 1.0 × 105 IU/kg feed (D60~D 1.0 × 105) vitamin D3. The results showed that weight gain and specific growth rate increased as the dietary vitamin D3 levels increased from 450 to 3300 IU/kg (P < 0.05). The fish fed with D1670 and D3300 diets had higher crude lipid and ash levels than the fish fed the D60 diet (P < 0.05). The fish fed D880, D1670, or D3300 diets had higher 25-OH-D3 and 1,25-(OH)2-D3 levels than the fish fed the D60 diet (P < 0.05). The fish fed D880, D1670, D3300, or D1.0 × 105 diets had higher osteocalcin levels than the fish fed the D60 diet (P < 0.05). Based on the broken line method analysis of weight gain and osteocalcin, the dietary vitamin D3 requirement of juvenile Siberian sturgeon was estimated to be 1683.30 and 1403.27 IU/kg per diet, respectively.

Isolation, Identification, and Optimization of Culture Conditions of a Bioflocculant-Producing Bacterium Bacillus megaterium SP1 and Its Application in Aquaculture Wastewater Treatment.

A bioflocculant-producing bacterium, Bacillus megaterium SP1, was isolated from biofloc in pond water and identified by using both 16S rDNA sequencing analysis and a Biolog GEN III MicroStation System. The optimal carbon and nitrogen sources for Bacillus megaterium SP1 were 20 g L-1 of glucose and 0.5 g L-1 of beef extract at 30°C and pH 7. The bioflocculant produced by strain SP1 under optimal culture conditions was applied into aquaculture wastewater treatment. The removal rates of chemical oxygen demand (COD), total ammonia nitrogen (TAN), and suspended solids (SS) in aquaculture wastewater reached 64, 63.61, and 83.8%, respectively. The volume of biofloc (FV) increased from 4.93 to 25.97 mL L-1. The addition of Bacillus megaterium SP1 in aquaculture wastewater could effectively improve aquaculture water quality, promote the formation of biofloc, and then form an efficient and healthy aquaculture model based on biofloc technology.