Implications on growth performance, glucose metabolism, PI3K/AKT pathway, intestinal flora induced by dietary taurine in a high-carbohydrate diet for grass carp (Ctenopharyngodon idella).

An 8-week experiment was performed to investigate the influence on growth performance, plasma biochemistry, glucose metabolism and the insulin pathway of supplementation of dietary taurine to a high-carbohydrate diet for grass carp. In this study, fish were fed diets at one of two carbohydrate levels, 31·49 % (positive control) or 38·61 % (T00). The high-carbohydrate basal diet (T00), without taurine, was supplemented with 0·05 % (T05), 0·10 % (T10), 0·15 % (T15) or 0·20 % (T20) taurine, resulting in six isonitrogenous (30·37 %) and isolipidic (2·37 %) experimental diets. The experimental results showed that optimal taurine level improved significantly weight gain, specific growth rate (SGR), feed utilisation, reduced plasma total cholesterol levels, TAG and promoted insulin-like growth factor level. Glucokinase, pyruvate kinase and phosphoenolpyruvate carboxykinase activities showed a quadratic function model with increasing dietary taurine level, while hexokinase, fatty acid synthetase activities exhibited a positive linear trend. Optimal taurine supplementation in high-carbohydrate diet upregulated insulin receptor (Ir), insulin receptor substrate (Irs1), phosphatidylinositol 3-kinase (pi3k), protein kinase B (akt1), glycogen synthase kinase 3 ß (gs3kß) mRNA level and downregulated insulin-like growth factor (igf-1), insulin-like growth factor 1 receptor (igf-1R) and Fork head transcription factor 1 (foxo1) mRNA level. The above results suggested that optimal taurine level could improve growth performance, hepatic capacity for glycolipid metabolism and insulin sensitivity, thus enhancing the utilisation of carbohydrates in grass carp. Based on SGR, dietary optimal tributyrin taurine supplementation in grass carp was estimated to be 0·08 %.

Muscular lipidomics and transcriptomics reveal the effects of bile acids on lipid metabolism in high-fat diet-fed grouper.

Little information is available on how exogenous bile acids alter lipid metabolism in muscle of fish. In the present study, an 8-week feeding trial were used to investigate the impacts of bile acids on lipid deposition, lipid metabolism, lipidomics, and transcriptomics in muscle of pearl gentian grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) fed a high-fat diet (HD). The HD treatment significantly increased the crude lipid content, while bile acids diet (BD) treatment decreased it (p = 0.057). BD treatment significantly decreased triglycerides level and significantly increased phosphatidylcholines, phosphatidylethanolamines, and phosphatidylglycerol levels. The contents of TG (17:0/18:2/18:2), TG (17:1/18:2/22:6), PC (6:0/22:1), PC (9:0/26:1), PC (26:1/6:0), PC (17:2/18:2), PE (16:0/18:1), PE (18:0/17:1), PG (18:0/20:5), PG (18:3/20:5), PG (19:0/16:1), and PG (18:0/18:1) in muscle were well response to dietary lipid level and bile acids supplementation. HD and BD groups induced a variety of adaptive metabolic responses in transcriptomics. HD treatment increased the lipogenesis and decreased lipolysis, whereas BD treatment decreased the lipogenesis and increased lipolysis. Present study revealed the improvement of muscular lipid metabolism and lipid composition in response to bile acids administration in pearl gentian grouper.

Glutamine improves growth and intestinal health in juvenile hybrid groupers fed high-dose glycinin.

Glutamine addition can improve immunity and intestinal development in fish. This study examined the protective roles of glutamine on growth suppression and enteritis induced by glycinin in juvenile hybrid groupers (female Epinephelus fuscoguttatus × male Epinephelus lanceolatus). The experiment set four isonitrogenous and isolipidic trial diets: a diet containing 10% glycinin (11S), 10% of 11S diet supplemented with 1% or 2% alanine-glutamine (1% or 2% Ala-Gln), and a diet containing neither 11S nor Ala-Gln (FM). A feeding trial was conducted in hybrid grouper for 8 weeks. Weight gain and specific growth rates in Groups 1% and 2% Ala-Gln were significantly higher than those of the 11S group but were similar to those of the FM group. The intestinal muscular layer thickness, plica height and width of the 2% Ala-Gln group were significantly higher than those of Group 11S. The enterocyte proliferation efficiency of the 11S group was significantly lower compared to other groups. Compared with the 11S group, Groups 1% and 2% Ala-Gln fish had increased intestinal lysozyme activities, complement 3 and immunoglobulin M as well as cathelicidin contents. The mRNA levels of tnf-α, il-1ß, ifn-α, and hsp70 genes were more downregulated in Groups 1% and 2% Ala-Gln than in Group 11S. Compared with FM group, fish from the 11S group had significantly lower mRNA levels of myd88, ikkß, and nf-κb p65 genes. These three values in the 2% Ala-Gln group were significantly lower than those in Group 11S but not significantly different from those of Group FM. The relative abundance of Vibrio in Group 11S was higher than that in Groups FM and 2% Ala-Gln. Intestinal glutamine, glutaminase, glutamic acid, α-ketoglutarate, malate dehydrogenase and ATP contents were higher in Groups 1% and 2% Ala-Gln than in Group 11S. These results suggest that glutamine is a useful feed additive to enhance growth and intestinal immunity, alleviate inflammation, and modulate gut microbiota in hybrid grouper fed high-dose glycinin.

Replacement of Dietary Fishmeal with Clostridium autoethanogenum Protein on Lipidomics and Lipid Metabolism in Muscle of Pearl Gentian Grouper.

Clostridium autoethanogenum protein (CAP) is an economical and alternative protein source. Here, three experimental diets were formulated with CAP replacing 0% (CAP-0), 30% (CAP-30), and 60% (CAP-60) of fishmeal to investigate the alterations of structure integrity, fatty acids profiles, and lipid metabolism in the muscle of pearl gentian grouper. With increasing levels of CAP substitution, the percentages of 16 : 0 or 18 : 0 were decreased in triglycerides (TG) and diacylglycerols (DG); 18 : 1 or 18 : 2 was increased at the sn-1 and sn-2 positions in phosphatidylethanolamines; 20 : 5n-3 was increased in TG and DG. The phosphatidylcholines (PC) (18 : 3/20 : 5), PC(22 : 6/17 : 1), and sphingomyelins (d19 : 0/24 : 4) were identified as potential lipid biomarkers between CAP treatments. The CAP-30 treatment enhanced lipolysis and lipogenesis, while the CAP-60 treatment inhibited lipogenesis. In conclusion, fishmeal replacement with CAP affected the lipid characteristics and lipid metabolism, whereas it did not affect the structural integrity and fatty acids profiles in the muscle of pearl gentian grouper.

Effects of Clostridium butyricum on Growth Performance, Intestinal Health, and Disease Resistance of Hybrid Grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) Fed with Cottonseed Protein Concentrate (CPC) Replacement of Fishmeal.

An 8-week feeding trial was conducted to investigate the effects of C. butyricum on the growth performance, microbiota, immunity response, and disease resistance in hybrid grouper fed with cottonseed protein concentrate (CPC) replacement of fishmeal. Six groups of isonitrogenous and isolipid diets were formulated including a positive control group (50% fishmeal, PC), a negative control group (CPC replaced 50% of fishmeal protein, NC), and Clostridium butyricum supplemented with 0.05% (C1, 5 × 108 CFU/kg), 0.2% (C2, 2 × 109 CFU/kg), 0.8% (C3, 8 × 109 CFU/kg), and 3.2% (C4, 3.2 × 1010 CFU/kg), respectively, to the NC group. The results showed that weight gain rate and specific growth rate were significantly higher in the C4 group than that in the NC group (P < 0.05). After supplementation with C. butyricum, the amylase, lipase, and trypsin activities were significantly higher than the NC group (P < 0.05; except group C1), and the same results were obtained for intestinal morphometry. The intestinal proinflammatory factors were significantly downregulated, and the anti-inflammatory factors were significantly upregulated in the C3 and C4 groups compared with the NC group after supplementation with 0.8%-3.2% C. butyricum (P < 0.05). At the phylum level, the PC, NC, and C4 groups were dominated by the Firmicutes and the Proteobacteria. At the genus level, the relative abundance of Bacillus in the NC group was lower than that in the PC and C4 groups. After supplementation with C. butyricum, grouper in the C4 group showed significantly higher resistance to V. harveyi than the NC group (P < 0.05). Above all, taking into account the effects of immunity and disease resistance, it was recommended to supplement 3.2% C. butyricum in the diet of grouper fed the replacement of 50% fishmeal protein by CPC.

Effect of black soldier fly (Hermetia illucens) larvae meal on lipid and glucose metabolism of Pacific white shrimp Litopenaeus vannamei.

The present study investigated the effect of black soldier fly (Hermetia illucens) larvae meal (BSF) on haemolymph biochemical indicators, muscle metabolites as well as the lipid and glucose metabolism of Pacific white shrimp Litopenaeus vannamei. Four diets were formulated in which the control diet contained 25 % of fishmeal (FM) and 10 % (BSF10), 20 % (BSF20), and 30 % (BSF30) of FM protein were replaced with BSF. Four hundred and eighty shrimp (0·88 ± 0·00 g) were distributed to four groups of three replicates and fed for 7 weeks. Results showed that growth performance of shrimp fed BSF30 significantly decreased compared with those fed FM, but there was no significant difference in survival among groups. The whole shrimp crude lipid content, haemolymph TAG and total cholesterol were decreased with the increasing BSF inclusion. The results of metabolomics showed that the metabolite patterns of shrimp fed different diets were altered, with significant changes in metabolites related to lipid metabolism, glucose metabolism as well as TCA cycle. The mRNA expressions of hk, pfk, pk, pepck, ampk, mcd, cpt-1 and scd1 in hepatopancreas were downregulated in shrimp fed BSF30, but mRNA expression of acc1 was upregulated. Unlike BSF30, the mRNA expressions of fas, cpt-1, fbp and 6pgd in hepatopancreas were upregulated in shrimp fed BSF20. This study indicates that BSF20 diet promoted lipid synthesis and lipolysis, while BSF30 diet weakened ß-oxidation and glycolysis as well as affected the unsaturated fatty acids synthesis, which may affect the growth performance and body composition of shrimp.

Effects of fishmeal replacement by black soldier fly on growth performance, digestive enzyme activity, intestine morphology, intestinal flora and immune response of pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

An 8-week feeding trial was conducted to investigate the influence of partial replacement of fishmeal (FM) by black soldier fly (BSF) (Hermetia illucens) on the growth, distal intestine morphology, intestinal flora, and intestinal immune response of pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Four diets were formulated, 0% (0 g kg-1), 10% (50 g kg-1), 20% (100 g kg-1) and 30% (150 g kg-1) fishmeal were replaced with BSF, named as FM, BSF10, BSF20, BSF30, severally. The study found that, with the increasing dietary BSF levels, growth and feed conversion ratio of fish decreased significantly (P < 0.05). Chitinase and trypsin activities were significantly increased with increasing dietary BSF levels (P < 0.05). With the increasing dietary BSF levels, distal intestinal muscularis thickness and mucosal fold length decreased significantly (P < 0.05), as well as total abundance of intestinal flora. The relative abundance of four phyla and six genera among the top 20 genera were significantly affected by dietary BSF levels (P < 0.05). With the increasing dietary BSF levels, the mRNA levels of nf-κbem1, r-cel and il-10 up-regulated significantly (P < 0.05). For fish fed BSF30 diet, the mRNA levels of myd88 and tlr22 were significantly higher than fish fed FM diet (P < 0.05). In conclusion, replacement fishmeal with BSF increased activity of digestive enzymes, but negatively affected growth performance and intestinal health of pearl gentian grouper.

Glycerol monolaurate (GML), a medium-chain fatty acid derivative, ameliorate growth performance, immune function, disease resistance and intestinal microbiota in juvenile hybrid grouper (Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂).

In order to evaluate the effects of glycerol monolaurate (GML) on the growth performance, immunology function, disease resistance and intestinal microbiota for hybrid groupers. Seven levels of GML (0, 600, 1200, 1800, 2400, 3000 and 3600 mg/kg) were added to diets and were noted as the G1 (control group), G2, G3, G4, G5, G6 and G7, respectively. Each experimental diet was fed to triplicate groups of 30 juvenile groupers for 8 weeks. The FBW, WGR and SGR were significantly higher and FCR was significantly lower in the G4 group compared to the G1 group (P < 0.05). Serum immune enzyme activities (ACP, AKP and LZM) rose and then fell and had the highest values in the G4 group (P < 0.05). The expression of TNF-α and IL6 in head kidney was significantly inhibited (P < 0.05), while the expression of TLR22 was increased (P < 0.05). After the Vibrio parahaemolyticus challenge test, ACP and AKP activities were increased in the G4 and G5 groups, while mortality was lower than in the G1 group (P < 0.05). GML significantly modulated the abundance of intestinal microbiota, with the G4 and G5 groups increasing the relative abundance of the Firmicutes and Bacillus, respectively (P < 0.05). The alpha diversity of the G5 group (Sob, Chao1 and ACE) was significantly higher than that of the G1 group (P < 0.05). In summary, the optimal level of GML was 1700 mg/kg according to the regression equation model fitted by the WGR index.

Effects of dietary chenodeoxycholic acid supplementation in a low fishmeal diet on growth performance, lipid metabolism, autophagy and intestinal health of Pacific white shrimp, Litopenaeus vannamei.

An 8-week feeding trial was conducted to evaluate the effects of chenodeoxycholic acid (CDCA) on growth performance, body composition, lipid metabolism, and intestinal health of juvenile white shrimp, Litopenaeus vannamei fed a low fishmeal diet. Four practical diets were formulated: HFM (25% fishmeal), LFM (15% fishmeal), LB1 (LFM + 0.04% CDCA), LB2 (LFM + 0.08% CDCA). Each diet was assigned to four tanks with forty shrimp (initial weight 0.33 ± 0.03 g) per tank. The results indicated that the growth performance of shrimp were similar between the four groups; the crude lipid content of shrimp fed the LB2 diet was significantly lower than those fed the HFM diet (P < 0.05). The lipase activity content in hepatopancreatic were significantly higher in the two CDCA supplemented groups than that in LFM group; the contents of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol in hemolymph were significantly lower in LFM group, LB1 group and LB2 group than that in HFM group (P < 0.05). The shrimp fed LB1 diet was significantly decreased the intestinal expression levels of tube than those fed in HFM diet; the intestinal gene expression of imd and toll were significantly lower in LB2 group than those in HFM group (P < 0.05). The results of hepatopancreas gene expression suggest that shrimp fed the LFM diet showed significantly upregulated expression levels of sterol regulatory element-binding protein (srebp), acetyl-CoA carboxylase (acc), and carnitine palmitoyltransferase 1 (cpt-1) than those fed the HFM diet; shrimp fed the LB1 diet showed significantly upregulated expression levels of srebp, acc, and AMP-activated protein kinase (ampk) than those fed the HFM diet; shrimp fed the LB2 diet had higher expression levels of srebp, acc, and cpt-1 than those fed the HFM diet (P < 0.05). In the hepatopancreas, the shrimp fed the LFM diet shown significantly up-regulated the expression levels of beclin1 compared to those fed HFM diet; the expression levels of autophagy-related protein13 (atg3), autophagy-related protein 12 (atg12) of in shrimp fed the LB1 diet were significantly higher than those fed the HFM diet; and the expression levels of autophagy-related protein13 (atg13), beclin1, atg3, atg12, autophagy-related protein 9 (atg9) of shrimp fed LB2 diet were significantly higher than those fed the HFM diet (P < 0.05). The atg3 in intestine of shrimp fed the LB2 diet were significantly higher than those fed the HFM diet (P < 0.05). Intestinal mucous fold were damaged, hepatic tubules were disorganized and B cells appeared to be swollen in LFM group. The fold height and width of shrimp fed the diets supplemented with CDCA increased significantly than those fed the LFM diet (P < 0.05), the hepatic tubules were neatly arranged, and R cells increased. In conclusion, supplementary CDCA in a low fishmeal diet promoted lipid metabolism, enhanced autophagy of shrimp, also improved the health of the intestine and hepatopancreas.

Dietary phosphorus supplementation in the diet of Pacific white shrimp (Litopenaeus vannamei) alleviated the adverse impacts caused by high Clostridium autoethanogenum protein.

The study evaluated the effects of dietary phosphorus supplementation on the fishmeal replacement with Clostridium autoethanogenum protein (CAP) in the diet of L. vannamei. Four isonitrogenous and isolipid diets were formulated: the PC diet contains 25% fishmeal, the NC, P1 and P2 diets were replaced 40% fishmeal with CAP and supplemented with 0, 0.8 and 1.6% NaH2PO4 respectively (equivalent to dietary phosphorus level of 0.96%, 1.12% and 1.27%). Sampling and V. parahaemolyticus challenge test were conducted after 50-day-feeding (initial shrimp weight 1.79 ± 0.02 g). The results showed that there were no significant differences in the growth performance of shrimp among the 4 groups. The expressions of dorsal in the gut were significantly lower in shrimp fed the P1 and P2 diets than shrimp fed the NC diet and the expression of peroxinectin in the gut was lower in shrimp fed the NC diet than others. The cumulative mortality of shrimp after V. parahaemolyticus challenge was significantly lower in shrimp fed the P2 diet than those fed the NC diet. After the challenge, genes expressions related to the prophenoloxidase activating system (proPO, lgbp, ppaf) were inhibited in the hepatopancreas of shrimp fed NC diet but activated in shrimp fed the P1 diet compared to those fed the PC diet. The AKP and T-AOC activities were higher in shrimp fed the P2 diet than those fed the other diets. The thickness of muscle layer of shrimp fed the P1 diet was thicker than that in the other groups, and significant stress damage happened in the midgut of the shrimp fed the NC diet. The abundance of Pseudoalteromonas, Haloferula and Ruegeria in shrimp fed the P1 diet was higher than those fed the other diets, while Vibrio in shrimp fed the P2 diet was higher than those fed the other diets. This indicated that a low fishmeal diet with dietary phosphorus level of 1.12% could improve the histology, enhance immune response, and increase the abundance of beneficial bacteria in the gut of shrimp. The low fishmeal diet with dietary phosphorus level of 1.27% could improve disease resistance and antioxidant capacity, but there was a possibility of damage to the gut histology as well as increasing abundance of Vibrio in the gut microbiota of shrimp.

Supplementing artemisinin positively influences growth, antioxidant capacity, immune response, gut health and disease resistance against Vibrio parahaemolyticus in Litopenaeus vannamei fed cottonseed protein concentrate meal diets.

Artemisinin (ART) is a kind of Chinese herbal medicine worth exploring, which obtains various physiological activities. In order to study the prebiotic effect of ART on Litopenaeus vannamei fed cottonseed protein concentrate meal diets, six groups of isonitrogenous and isolipid diets were prepared (including the fish meal control group, FM; cottonseed protein concentrate replacing 30% fishmeal protein and supplementing ART groups: ART0, ART0.3, ART0.6, ART0.9, and ART1.2). The feeding trials was lasted for 56 days. The results showed that the final body weight, weight gain and specific growth rate of the ART0.6 group were the highest, yet the feed coefficient rate of the ART0.6 group was the lowest significantly (P < 0.05). There was no significant difference in survival rate among treatments (P > 0.05). In serum, the content of malondialdehyde in ART0 group was the highest (P < 0.05); the activities of superoxide dismutase, catalase, phenol oxidase and lysozyme increased firstly and then decreased among the ARTs groups (P < 0.05). The activities of intestinal digestive enzymes (including the trypsin, lipase and amylase) showed an upward trend among the ARTs groups (P < 0.05). The histological sections showed that the intestinal muscle thickness, fold height and fold width in the FM group were significantly better than those in the ART0 group; while the mentioned above morphological indexes in the ART0 group were significantly lowest among the ARTs groups (P < 0.05). Sequencing of intestinal microbiota suggested that the microbial richness indexes firstly increased and then decreased (P < 0.05); the bacterial community structure of each treatment group was almost close; the relative abundance of pathogenic bacteria decreased significantly (P < 0.05), such as the Proteobacteria and Cyanobacteria at phylum level, besides the Vibrio and Candidatus Bacilloplasma at genus level. In intestinal tissue, the relative expression levels of TOLL1, TRAF6 and Pehaeidih3 showed up-regulated trends, while the expression of Crustin and LZM firstly up-regulated and then down-regulated (P < 0.05). The challenge experiment suggested that the cumulative mortality of FM group was significantly lower than that of ART0 group; besides the cumulative mortality firstly increased and then decreased between the ARTs groups (P < 0.05). In conclusion, the dietary supplementation of ART can improve the growth, antioxidant capacity, immune response, gut health and disease resistance of the shrimp. To be considered as a dietary immune enhancer, the recommended supplementation level of ART in shrimp's cottonseed protein concentrate meal diets is 0.43%.

Effects of the Clostridium butyricum on growth performance, antioxidant capacity, immunity and disease resistance of Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal in diet.

Clostridium butyricum (CB) is a gram-positive bacterium that secretes short-chain fatty acids such as butyric acid and so on. An 8-week feeding trial was conducted to investigate the effects of CB on the growth performance, antioxidant capacity, immunity and resistance to Vibrio parahaemolyticus in Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal. Six iso-nitrogenous (40%) and iso-lipidic (6%) diets were formulated including a positive control group (PC, 25% fishmeal), a negative control group (NC, CPC replaced 30% of fishmeal protein), and 0.03% (C1, 3 × 108 CFU/kg), 0.12% (C2, 1.2 × 109 CFU/kg), 0.48% (C3, 4.8 × 109 CFU/kg) and 1.92% (C4, 1.92 × 1010 CFU/kg) CB were supplemented on the negative control group (NC). After the feeding trial, the remaining shrimp in each treatment group were subjected to a challenge experiment with Vibrio parahaemolyticus. The results indicated that weight gain rate (WGR), specific growth rate (SGR) in C4 group were significantly lower than those in PC and C2 groups (P < 0.05); the feed conversion ratio (FCR) was significantly higher than that of PC and C2 groups (P < 0.05). There was no significant difference in survival rate (SR) among all groups (P > 0.05). Compared to the PC and NC groups, the total superoxide capacity, superoxide dismutase and lysozyme were significantly higher in the C4 group (P < 0.05); the glutathione peroxidase, acid phosphatase and alkaline phosphatase were significantly higher in the C3 group (P < 0.05); and the malondialdehyde was significantly lower in the C4 group (P < 0.05). The relative mRNA expressions of Toll receptor (TLR), innate immune deficiency gene (IMD), penaiedin3a (Pen3) were significantly down-regulated in the NC group than those in the PC group (P < 0.05). In addition, the relative mRNA expressions of TLR, IMD and Pen3 were significantly up-regulated in all groups supplemented with CB than those in the NC group (P < 0.05). Moreover, the cumulative mortality rate in the NC group was not significantly different from the PC group (P > 0.05) and was significantly higher than those in the C3 and C4 groups (P < 0.05). In conclusion, the CB supplementation on the basis of CPC replacement of 30% fishmeal protein enhanced significantly the antioxidant capacity, immunity and disease resistance of shrimp and improved its growth performance. Therefore, considering the factors of the growth, immunity and disease resistance, the CB supplementation of 0.12%-0.48% (1.2 × 109 CFU/kg-4.8 × 109 CFU/kg) was recommended in the diet of L. vannamei based on the results of this experiment.

Effects of Lysophospholipid Supplementation in Feed with Low Protein or Lipid on Growth Performance, Lipid Metabolism, and Intestinal Flora of Largemouth Bass (Micropterus salmoides).

The largemouth bass (Micropterus salmoides) were fed diets with three experimental feeds, a control diet (Control, crude protein (CP): 54.52%, crude lipid (CL): 11.45%), a low-protein diet with lysophospholipid (LP-Ly, CP: 52.46%, CL: 11.36%), and a low-lipid diet with lysophospholipid (LL-Ly, CP: 54.43%, CL: 10.19%), respectively. The LP-Ly and LL-Ly groups represented the addition of 1 g/kg of lysophospholipids in the low-protein and low-lipid groups, respectively. After a 64-day feeding trial, the experimental results showed that the growth performance, hepatosomatic index, and viscerosomatic index of largemouth bass in both the LP-Ly and LL-Ly groups were not significantly different compared to those in the Control group (P > 0.05). The condition factor and CP content of whole fish were significantly higher in the LP-Ly group than those in the Control group (P < 0.05). Compared with the Control group, the serum total cholesterol level and alanine aminotransferase enzyme activity were significantly lower in both the LP-Ly group and the LL-Ly group (P < 0.05). The protease and lipase activities in the liver and intestine of both group LL-Ly and group LP-Ly were significantly higher than those of the Control group (P < 0.05). Compared to both the LL-Ly group and the LP-Ly group, significantly lower liver enzyme activities and gene expression of fatty acid synthase, hormone-sensitive lipase, and carnitine palmitoyltransferase 1 were found in the Control group (P < 0.05). The addition of lysophospholipids increased the abundance of beneficial bacteria (Cetobacterium and Acinetobacter) and decreased the abundance of harmful bacteria (Mycoplasma) in the intestinal flora. In conclusion, the supplementation of lysophospholipids in low-protein or low-lipid diets had no negative effect on the growth performance of largemouth bass, but increased the activity of intestinal digestive enzymes, enhanced the hepatic lipid metabolism, promoted the protein deposition, and regulated the structure and diversity of the intestinal flora.

Evaluation of Six Novel Protein Sources on Apparent Digestibility in Pacific White Shrimp, Litopenaeus vannamei.

This study is aimed at evaluating the apparent digestibility coefficients (ADC) of six novel protein sources in Pacific white shrimp (Litopenaeus vannamei), including black soldier fly larvae meal (BSFLM), Chlorella vulgaris meal (CM), cottonseed protein concentrate (CPC), Tenebrio molitor meal (TM), Clostridium autoethanogenum protein (CAP), and methanotroph (Methylococcus capsulatus, Bath) bacteria meal (BPM). The control diet (CD) was formulated to contain 448.8 g/kg crude protein and 71.8 g/kg crude lipid. Then, six experimental diets were formulated to contain 70% CD and 30% test ingredients. The yttrium oxide was used as an exogenous indicator for apparent digestibility detection. Six hundred and thirty healthy and uniform-sized shrimp (approximately 3.04 ± 0.01 g) were randomly distributed into triplicate groups of 30 shrimp and they were fed three times daily. After the shrimp was acclimating for one week, their feces were collected 2 hours after the morning feeding until sufficient samples were available for compositional analysis to calculate apparent digestibility. The apparent digestibility coefficients for a dry matter of diets (ADCD) and ingredients (ADCI) as well as the apparent digestibility coefficients for crude protein (ADCPro), crude lipid (ADCL), and phosphorus (ADCP) of test ingredients were calculated. Results showed that the growth performance of shrimp fed BSFLM, TM, and BPM diets significantly decreased compared to that fed the CD (P < 0.05), and no significant differences were found among those fed CD, CM, CAP, and CPC diets (P > 0.05). There were no significant differences in survival among each group (P > 0.05). As for the diets, results showed that the ADCD of BSFLM, CM, CPC, and TM diets was significantly lower than that of CD, while that of the CAP diet was significantly higher than that of CD (P < 0.05) and there were no significant differences between BPM and CD diets (P > 0.05). As for the test ingredients, the ADCPro and ADCL of BSFLM, CM, CPC, and TM were significantly lower than those of CD in Litopenaeus vannamei (P < 0.05). The ADCPro of CAP was significantly higher than that of CD (P < 0.05), but no significant differences were found in ADCL between CAP and CD (P > 0.05). The ADCPro of BPM was significantly lower than that of CD (P < 0.05), but there were no significant differences in ADCL between BPM and CD (P > 0.05). The ADCP of CM, CAP, and BPM were significantly higher than that of CD, while that of BSFLM was significantly lower than that of CD (P < 0.05), and no significant differences were found in ADCP between TM and CD (P > 0.05). To conclude, newly developed protein sources such as single-cell protein (CAP, BPM, and CM) showed great potential as a fishmeal alternative, and insect protein meals (TM and BSFLM) were less effective for shrimp compared to the CD. Although the utilization of CPC by shrimp was lower than other protein sources, it had been much improved compared to the untreated cottonseed meal. The present study will contribute to the application of novel protein sources in shrimp feeds.

Farnesoid X receptor regulates PI3K/AKT/mTOR signaling pathway, lipid metabolism, and immune response in hybrid grouper.

Some diseases related to lipid metabolism increase yearly in cultured fish, and the farnesoid X receptor (FXR) is a nuclear protein that plays a key role in inflammatory responses and lipid metabolism. However, the roles of FXR in hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) remain poorly understood. The main objective of this study was to explore the roles of hepatic FXR in triggering the immune response and the potential functions of FXR in regulating the lipid metabolism. In the present study, the full-length sequence of fxr from hybrid grouper was cloned and characterized for the first time. Upon the Vibrio parahaemolyticus stimulation, the transcriptional level of fxr was rapidly elevated in the head kidney tissue in the early stage of infection. In vivo and vitro, activation of FXR by obeticholic acid (OA) significantly increased the concentrations and mRNA levels of hepatic inflammatory cytokines. These effects were inversed when FXR was inhibited by guggulsterone (GU). Moreover, the activation of FXR to suppress the PI3K/AKT/mTOR signaling pathway improves hepatic lipid metabolism and reduces hepatic lipid accumulation in vivo and vitro. In addition, the inhibition of FXR activated the PI3K/AKT/mTOR pathway, decreased the lipolysis and increased the lipogenesis, and subsequently increased the lipid accumulation in fish. These results revealed the positive roles of FXR in triggering immune responses and improving lipid metabolism and accumulation in hybrid grouper.

Dietary taurine stimulates the hepatic biosynthesis of both bile acids and cholesterol in the marine teleost, tiger puffer (Takifugu rubripes).

Taurine (TAU) plays important roles in the metabolism of bile acids, cholesterol and lipids. However, little relevant information has been available in fish where TAU has been identified as a conditionally essential nutrient. The present study aimed to investigate the effects of dietary TAU on the metabolism of bile acids, cholesterol and lipids in tiger puffer, which is both an important aquaculture species and a good research model, having a unique lipid storage pattern. An 8-week feeding trial was conducted in a flow-through seawater system. Three experimental diets differed only in TAU level, that is, 1·7, 8·2 and 14·0 mg/kg. TAU supplementation increased the total bile acid content in liver but decreased the content in serum. TAU supplementation also increased the contents of total cholesterol and HDL-cholesterol in both liver and serum. The hepatic bile acid profile mainly includes taurocholic acid (94·48 %), taurochenodeoxycholic acid (4·17 %) and taurodeoxycholic acid (1·35 %), and the contents of all these conjugated bile acids were increased by dietary TAU. The hepatic lipidomics analysis showed that TAU tended to decrease the abundance of individual phospholipids and increase those of some individual TAG and ceramides. The hepatic mRNA expression study showed that TAU stimulated the biosynthesis of both bile acids and cholesterol, possibly via regulation of farnesoid X receptor and HDL metabolism. TAU also stimulated the hepatic expression of lipogenic genes. In conclusion, dietary TAU stimulated the hepatic biosynthesis of both bile acids and cholesterol and tended to regulate lipid metabolism in multiple ways.

The characteristics of glucose homoeostasis in grass carp and Chinese longsnout catfish after oral starch administration: a comparative study between herbivorous and carnivorous species of fish.

An oral starch administration trial was used to evaluate glucose homoeostasis in grass carp (Ctenopharyngodon idella) and Chinese longsnout catfish (Leiocassis longirostris Günther). Fish were administered with 3 g of a water and starch mixture (with 3:2 ratio) per 100 g body weight after fasting for 48 h. Fish were sampled at 0, 1, 3, 6, 12, 24 and 48 h after oral starch administration. In grass carp, plasma levels of glucose peaked at 3 h but returned to baseline at 6 h. However, in Chinese longsnout catfish, plasma glucose levels peaked at 6 h and returned to baseline at 48 h. The activity of intestinal amylase was increased in grass carp at 1 and 3 h, but no significant change in Chinese longsnout catfish was observed. The activity of hepatic glucose-6-phosphatase fell significantly in grass carp but change was not evident in Chinese longsnout catfish. The expression levels and enzymic activity of hepatic pyruvate kinase increased in grass carp, but no significant changes were observed in the Chinese longsnout catfish. Glycogen synthase (gys) and glycogen phosphorylase (gp) were induced in grass carp. However, there was no significant change in gys and a clear down-regulation of gp in Chinese longsnout catfish. In brief, compared with Chinese longsnout catfish, grass carp exhibited a rapid increase and faster clearance rate of plasma glucose. This effect was closely related to significantly enhanced levels of digestion, glycolysis, glycogen metabolism and glucose-induced lipogenesis in grass carp, as well as the inhibition of gluconeogenesis.

Effects of dietary n-3 highly unsaturated fatty acids on growth, non-specific immunity, expression of some immune-related genes and resistance to Vibrio harveyi in hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ Epinephelus lanceolatu).

This study was conducted to investigate the effects of dietary n-3 highly unsaturated fatty acids (n-3 HUFA) on growth performance, non-specific immunity, expression of some immune-related genes and resistance to Vibrio harveyi in juvenile hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ Epinephelus lanceolatu). Six isoproteic and isolipidic experimental diets were formulated with graded levels of n-3 HUFA (0.65, 1.00, 1.35, 1.70, 2.05 and 2.40% of dry matter, respectively), and the 0.65% group was used as control group. Each diet was randomly allocated to triplicate groups of fish in 1000 L fiberglass tank, and each tank was stocked with 40 fish (initial weight 12.06 ± 0.01 g) for 8 weeks. Results showed that feed conversion ratio (FCR), survival rate (SR), hepatosomatic index (HSI) and condition factor (CF) were all not significantly affected by dietary n-3 HUFA levels (P > 0.05). Weight gain (WG) and specific growth rate (SGR) in 1.35% group were significantly higher than those in 2.40% group (P < 0.05). Crude lipid of body in 1.00% group was significantly lower than that in 1.70% and 2.40% groups (P < 0.05). Liver and muscle fatty acid profiles reflected that of diets. Before challenge with Vibrio harveyi, the activity of serum superoxide dismutase (SOD), catalase (CAT) and content of complement 3 (C3) in 1.35% and 1.70% groups significantly higher than those of control group (P < 0.05). After challenge with Vibrio harveyi, serum CAT, glutathione peroxidase (GSH-PX), lysozyme (LZM) and C3 all increased sharply, while SOD showed the opposite trend. Before challenge with Vibrio harveyi, the expression levels of intestine toll-like receptor 22 (TLR22) and myeloid differentiation factor 88 (MyD88) mRNA in 2.40% group were significantly increased, and the expression levels of tumour necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) mRNA in 2.05% group were significantly higher than those in 1.00% and 1.35% groups (P < 0.05). In addition, the TLR22 and IL-1ß mRNA levels in kidney of 1.70% group were significantly lower than those in control group (P < 0.05). After challenge with Vibrio harveyi, the expression level of MyD88 mRNA in intestine of 1.35% group was significantly higher than that in 1.00% group and from 1.70% to 2.40% groups (P < 0.05), while TNF-α and IL-1ß obtained minimum values in 1.70% group. In the kidney, the interleukin 10 (IL10) mRNA expression was significantly higher in 1.70% group than that in other groups, while the IL-1ß expression in 1.70% group was on the contrary and significantly lower than that in 2.40% group (P < 0.05). Results of this study suggested that moderate dietary n-3 HUFA (1.47%-1.70% HUFA) could improve the growth performance, non-specific immunity and inhibit the inflammatory response of hybrid grouper.

The effect of partial replacement of fish meal by soy protein concentrate on growth performance, immune responses, gut morphology and intestinal inflammation for juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

An 8-week feeding trial was conducted to investigate the effects of partial replacement of fish meal by soy protein concentrate (SPC) on the growth performance, immune responses, intestine morphology and relation gene expression of intestinal inflammation for juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) (initial weight 12.5 ±â€¯0.00 g). Eight isonitrogenous and isolipidic diets (48.61% protein and 11.17% lipid) were formulated by replacing 0% (the control), 11%, 22%, 33%, 44%, 55%, 66%, and 77% of fish meal (FM) with SPC, respectively (the eight dietary be named FM, S11, S22, S33, S44, S55, S66, and S77, respectively). With the replacement level increased, the final body weight, weight gain ratio (WGR), specific growth rate (SGR), and survival rate of fish were significantly decreased (P < 0.05) compared with the group FM. By contrast, the feed conversion ratio (FCR) of fish was significantly increased (P < 0.05) when the replacement level up to 44%. Partial FM replacement by SPC (ranging from 11% to 77%) substantially reduced (P < 0.05) the serum total protein, albumin, and total cholesterol contents compared with the group FM. Liver total superoxide dismutase, glutathione peroxidase, catalase activities, and total antioxidant capacity showed the same trend of gradual increase first and then decrease. Their highest values were found in the replacement levels of 55%, 33%, 22%, and 55% and were significantly higher (P < 0.05) than the control group. The lowest malondialdehyde content was observed in group S77 and was significantly lower (P < 0.05) than that of the control group. The complements C3 and C4 contents of fish fed with experimental diets (replacement level ranged from 11% to 66%) were significantly higher (P < 0.05) than the group FM. The liver lysozyme activity of the control group was the lowest and was significantly lower than that of other dietary treatments (P < 0.05). Villus length and muscle thickness in the intestine of fish were significantly lower (P < 0.05) than other groups when the replacement level exceeded 44%. With dietary replacement levels increased, the TLR22, MyD88, p65, pro-inflammatory cytokines (IL-1ß, TNF-α, IL-12P40 and INF-γ) and anti-inflammatory cytokines (TGF-ß, IL-10, epinecidin, MHCIIß and hepcidin) mRNA levels in the proximal intestine were significantly up-regulated (P < 0.05). The TLR22, MyD88, p65, pro-inflammatory cytokines (IL-1ß, TNF-α, IL-12P40 and INF-γ) and anti-inflammatory cytokines (TGF-ß, IL-10, MHCIIß and hepcidin) mRNA levels in the mid intestine were significantly up-regulated (P < 0.05). The mRNA levels of TLR22, anti-inflammatory cytokines (IL-1ß, TNF-α, IL-12P40, INF-γ) and anti-inflammatory cytokines (TGF-ß, IL-10, epinecidin, MHCIIß and hepcidin) in the distal intestine were significantly down-regulated (P < 0.05). The mRNA expression of MyD88 and p65 mRNA were showed a tend increased first and then decreased, and the highest values were observed in group S33 and S55 (P < 0.05), respectively. Based on the present work, the correlation between WGR and FM replacement level with SPC was described using the broken-line model, which estimated the optimum FM replacement to 37.23% for juvenile hybrid grouper dietary.

Preliminary study of mechanisms of intestinal inflammation induced by plant proteins in juvenile hybrid groupers (♀Epinephelus fuscoguttatus×♂E. lanceolatu).

Fish fed a high plant protein diet exhibit intestinal inflammation, the mechanism of which needs to be clarified. We preliminarily elucidate the mechanism of the TLRs/MyD88-PI3K/Akt signalling pathway in intestinal inflammation induced by plant proteins. The diets contained 60% fish meal (FM, controls), or had 45% of the fish meal protein replaced by soybean meal (SBM), peanut meal (PM), cottonseed meal (CSM) or cottonseed protein concentrate (CPC). After an 8-week feeding trial, fish were challenged by injection of Vibrio parahaemolyticus bacteria for 7 days until the fish stabilized. The results showed that the specific growth rate (SGR) of the FM group was higher than other groups. The SGR of the CPC group was higher than those of the SBM, PM and CSM groups. The catalase (CAT) contents in the serum of fish fed a plant protein diet were higher than in FM fish. The abundances of Rhodobacteraceae and Microbacteriaceae in the MI (mid intestine) were higher in the CPC group. The TLR-2 expressions in the MI and DI of plant protein-fed fish were up-regulated. The expressions of IL-6 in the PI and MI, of hepcidin and TLR-3 in the MI, and of TLR-3 in the DI, were all lower than those of fish fed FM. In the PI, MI and DI, the protein expressions of P-PI3K/T-PI3K in the SBM and PM groups were higher than in the FM group. After the challenge, the cumulative mortalities in the FM and CPC groups were lower than those of the SBM, PM and CSM groups. These results suggested that plant protein diets reduced antioxidant capacity and glycolipid metabolism, hindered the development of the intestine and reduced intestinal flora diversity. TLR-3 is involved in the immune regulation of the PI in CPC group, MI and DI in SBM, PM, CSM and CPC groups, while might be involved in the immune regulation of the PI in SBM, PM and CSM groups. Furthermore, PI3K/Akt signaling does not participate in the regulation of PI and MI in the CSM group, MI and DI in the CPC group.