Excessive level of dietary insect protein negatively changed growth metabolomic and transcriptomic profiles of largemouth bass (Micropterus salmoides).

Hermetia illucens (HI) meal is a promising substitute for fish meal (FM) in the feeds of farmed fish. However, the impacts of dietary HI meal on largemouth bass (LMB) remain unknown. In this study, we formulated three isonitrogenous and isolipid diets with 0% (HI0, control), 20% (HI20) and 40% (HI40) of FM substituted by HI meal. A total of 270 juvenile largemouth bass with an initial body weight of 10.02 ± 0.03 g were used (30 fish per tank). After an 80-day feeding trial, the fish fed with the HI40 diet demonstrated decreased growth performance and protein efficiency ratio (PER), and increased liver oxidative indices and lipid accumulation compared to the control (P < 0.05). Transcriptomic analysis revealed the effects of high dietary HI meal on liver gene expression. Consistent with the reduced growth and disturbed liver oxidative status, the upregulated genes were enriched in the biological processes associated with protein catabolism and endoplasmic reticulum (ER) stress; while the downregulated genes were enriched in cellular proliferation, growth, metabolism, immunity and maintenance of tissue homeostasis. Differential metabolites in the liver samples were also identified by untargeted metabolomic assay. The results of joint transcriptomic-metabolomic analyses revealed that the pathways such as one carbon pool by folate, propanoate metabolism and alpha-linolenic acid metabolism were disturbed by high dietary HI meal. In summary, our data revealed the candidate genes, metabolites and biological pathways that account for the adverse effects of high HI meal diet on the growth and health of LMB.

Biochemical, histological and transcriptomic analyses for the immunological organs provide insights into heat stress-induced disease susceptibility in Largemouth Bass.

Aquaculture of Largemouth Bass (LMB, Micropterus salmoides), an economically important species, is badly affected by the outbreak of bacterial diseases in summer. However, the mechanisms underlying heat-induced disease susceptibility remain largely unknown. In this study, after exposure to 34 °C for 1, 7 and 14 d, the head kidney, spleen and blood of LMB were sampled for biochemical and histological assays to explore the effects of heat exposure on the oxidative and immunological indices. Compared to the controls maintained at 28 °C, chronic heat exposure (34 °C for 14 d) induced oxidative stress, caused cell apoptosis and decreased expression of the immunological genes in the head kidney and spleen tissues; and attenuated the blood immunological indices. Consistent with the impaired immunological functions, chronic heat exposure predisposed LMB to Aeromonas hydrophila infection and significantly (p < 0.001) increased tissue bacterial load. Furthermore, the effects of chronic heat exposure (heat), A. hydrophila infection (infection) and heat exposure followed by A. hydrophila infection (heat + infection) on gene expression in the head kidney and spleen of LMB were characterized by RNA sequencing. The results indicated that chronic heat exposure facilitated the bacteria-elicited changes in expression of the genes involved in a couple of metabolic and signaling pathways in both tissues. Upon heat + infection, the pathways involved in energy production and nutrients biosynthesis were enhanced, whereas those associated with the host cell functions such as cell-cell interactions and cell signaling were depressed. Our data provide new insights into the mechanisms underlying heat-induced disease susceptibility in LMB.

Effects of Yellow Mealworm (Tenebrio molitor) on Growth Performance, Hepatic Health and Digestibility in Juvenile Largemouth Bass (Micropterus salmoides).

This study investigated the effects of yellow mealworm meal (TM) on growth performance, hepatic health and digestibility in juvenile largemouth bass (Micropterus salmoides). The fish were fed with the basic feed and the test feed (70% basic feed and 30% raw materials) containing Cr2O3, and feces were collected for digestibility determination. The fish were fed with five isonitrogenous (47% crude protein) and isolipidic (13% crude lipid) diets, in which fishmeal (FM) was replaced with 0% (TM0), 12% (TM12), 24% (TM24), 36% (TM36) and 48% (TM48) TM. The fish were reared in cylindrical plastic tanks in a recirculating aquaculture system for 11 weeks. The apparent digestibility coefficients (ADC), of dry matter, crude protein and crude lipid, in largemouth bass of TM were 74.66%, 91.03% and 90.91%, respectively. The ADC of total amino acid (TAA) of TM in largemouth bass was 92.89%, and the ADC of essential amino acid (EAA) in TM in largemouth bass was 93.86%. The final body weight (FBW), weight gain rate (WGR) and specific growth rate (SGR) in the TM24 group were significantly higher than those in other groups. Similarly, the highest mRNA expression levels of hepatic protein metabolism genes (pi3k, mtor, 4ebp2 and got) and antioxidant enzyme (glutathione peroxidase, Gpx; catalase, Cat) activities were observed in the TM24 group. Moreover, the expression levels of anti-inflammatory factors (il-10 and tgf) in liver were up-regulated and the expression levels of pro-inflammatory factors (il-8 and il-1ß) in liver were down-regulated. Quadratic regression model analysis, based on weight gain rate (WGR) against dietary TM level, indicated that the optimum level of dietary TM replacing FM in largemouth bass diet was 19.52%. Appropriate replacement levels (less than 36%) of FM by TM in the diets can enhance the antioxidant capacity and immunity of largemouth bass. However, high levels of FM substitution with TM (more than 48%) in the feeds can damage the liver health and inhibit the growth of largemouth bass. Notably, largemouth bass has high ADC and high utilization of TM, which indicates that it is feasible to use TM as feed protein source for largemouth bass.

Transcriptome analysis provides insights into the molecular mechanism of liver inflammation and apoptosis in juvenile largemouth bass Micropterus salmoides fed low protein high starch diets.

The present study was conducted to investigate the regulatory mechanism of liver injury in largemouth bass Micropterus salmoides (LMB) fed low protein high starch diets. Two isolipidic and isoenergetic diets were formulated with different protein and starch ratios, being named as diets P49S9 (48.8 % protein and 9.06 % starch) and P42S18 (42.4 % protein and 18.2 % starch). Each diet was fed to triplicate replicates of LMB (initial body weight, 4.65 ± 0.01 g) juveniles. Fish were fed to visual satiation for 8 weeks. The results indicated that though the P42S18 fish up-regulated the feeding ratio to meet their protein requirements, feeding efficiency ratio and growth performance were impaired in treatment P42S18 as compared to treatment P49S9. Periodic acid-Schiff (PAS) staining showed glycogen accumulated in the liver of LMB fed low protein high starch diets, and the reason should be attributed to down-regulated expression of the glycogenolytic glycogen debranching enzyme. Lower liver lipid level was associated with feeding low protein high starch diets in LMB, which should be resulted from the changes in hepatic glycerolipid metabolism regulated by lipoprotein lipase (representative of triglyceride synthesis, up-regulated) and diacylglycerol acyltransferase (representative of triglyceride breakdown, down-regulated). Though fasting plasma glucose level was comparable, treatment P42S18 performed inferior glucose tolerance to treatment P49S9. Hematoxylin-eosin (HE) and TdT-mediated dUTP Nick-End Labeling (TUNEL) staining suggested that feeding low protein high starch diets induced disruption of structural integrity, inflammation and apoptosis in the hepatocytes of LMB. As expected, KEGG pathways analysis indicated that many of the up-regulated differentially expressed genes were enriched in AGE (advanced glycation end product)/RAGE (receptor for AGE), Toll-like receptor and apoptosis signaling pathways. Our transcriptome data revealed that feeding low protein high starch diets might promote the accumulation of AGEs in LMB, which bound to RAGE and subsequently induced PI3K/Akt signal pathway. The activation of Akt induced NF-κB translocation into the nucleus thus releasing proinflammatory factors including tumor necrosis factor-α (TNF-α) and interleukin-8. The release of these inflammatory factors concomitantly induced T cell stimulation and natural killer cells chemotactic effects through Toll-like receptor signaling pathway. Besides mediating inflammation and immune response, TNF-α signal transduction participated in mediating apoptosis through the receptor of TNF (TNF-R1) pathway by up-regulating the expression of caspase 8 and cytochrome c. In conclusion, our results demonstrated that feeding low protein and high starch diets induced hepatocytes inflammation and apoptosis in LMB through the PI3K/Akt/NF-κB signaling pathway.

Yeast culture improved the growth performance, liver function, intestinal barrier and microbiota of juvenile largemouth bass (Micropterus salmoides) fed high-starch diet.

The present study was conducted to investigate the effects of yeast culture on the growth, health and microflora of the juvenile largemouth bass fed high-starch diet. The experiment set three isonitrogenous and isolipidic diets, control (high-starch diet), HSY1 (high-starch diet with 1% yeast culture) and HSY3 (high-starch diet with 3% yeast culture). A feeding trial was conducted in largemouth bass juveniles for 8 weeks. The results indicated fish fed with 3% yeast culture not only could improve specific growth rate (SGR), but also significantly decreased hepatic lipid content, hepatic glycogen content, and hepatopancreas somatic index (HSI) compared with the control group (p<0.05). The total superoxide dismutase (T-SOD) and catalase (CAT) activities of HSY3 group significantly increased while malondialdehyde (MDA) content significantly reduced in liver compared with the control group (p<0.05). Meanwhile, the mRNA expression levels of hepatic Sod and Cat were up-regulated (p<0.05), and liver metabolism showed 111 metabolites were significantly changed in HSY3 group, liver lipid metabolism pathway remarkably changed. Besides, the intestinal anti-inflammatory cytokines were significantly up-regulated, and the pro-inflammatory cytokines were significantly down-regulated as the inclusion of yeast culture (p<0.05). Notably, HSY3 group diet up-regulated the expression of Zo-1, Claudin and Occludin in intestine compared with the other groups (p<0.05). Serum d-lactate (D-lac), diamine oxidase (DAO) and lipopolysaccharide (LPS) decreased significantly with the inclusion of yeast culture (p<0.05). Furthermore, the abundance of probiotics (such as Lactobacillus, Bacillus and Bifidobacterium) increased significantly, and the abundance of intestinal potential pathogenic bacteria (Plesiomonas) decreased in HSY3 group (p<0.05). The phenotypic analysis showed that gram-negative bacteria significantly decreased while gram-positive bacteria increased in HSY3 group (p<0.05). All in all, this study revealed that supplementation of 3% yeast culture can improve the growth performance and the health of juvenile largemouth bass, and has the potential to be used as an effective synbiotics for M. salmoides.

Intestinal morphology, immunity and microbiota response to dietary fibers in largemouth bass, Micropterus salmoide.

This study is aimed at identifying the effects of dietary fiber on gut health, as well as the association between that understanding and fiber consumption in fish. A total of 300 juvenile largemouth bass (micropterus salmoides, initial average weight: 15.38 ± 0.16g) were randomly divided into three treatment groups (4 replicates per group). Fish were fed with isoproteic and isolipidic diets containing 0% (low fiber, LF), 4% (moderate fiber, MF) and 8% (high fiber, HF) soybean fiber, respectively. The intestine and intestinal content of test fish per treatment group after 56 days of treatment were sampled. The results showed that the anterior intestinal sections had normal histological architecture, and no considerable damage or inflammation was observed in any histological section from all subjects examined. Curiously, fish fed the MF diet had better histological alterations than the other treatments. Meanwhile, the intestinal antioxidant capacity in the MF group was significantly promoted when compared to the other groups, as well as up-regulated expression of antioxidant-related genes including sod, cat and gpx with increasing dietary fiber concentrations. Importantly, the administrations of MF diet remarkably elevated largemouth bass innate immune parameters include intestinal inducible nitric oxide synthase (iNOS) activity, nitric oxide (NO) and total protein content. Similarly, dietary administrations of fiber down-regulated notablely the expression of pro-inflammatory cytokines including IL-8, IL-1ß and TNFα, whereas up-regulated tolerogenic cytokine IL-10 and TGF-ß1 mRNA levels. In addition, dietary fibers also modulated the community structure of the intestinal microbiota by significantly altering bacterial diversity. Dietary supplemental fibers regulated intestinal microbiota in largemouth bass, characterized by a reduced abundance of Fusobacteria along with increased abundances of Proteobacteria and Firmicutes. Taken together, the present results suggested that moderate fiber supplementation was beneficial to promoting intestinal health status of fish through antioxidant and anti-inflammatory effects, which could be at least partially responsible by the modulation of gut microbial composition.

Effects of supplemental dietary bile acids on growth, liver function and immunity of juvenile largemouth bass（Micropterus salmoides）fed high-starch diet.

The present study was conducted to investigate the effects of bile acids (BAs) on the growth, liver function and immunity of the largemouth bass fed high-starch diet. The experiment set three isonitrogenous and isoenergetic semi-purified diets, LS: low-starch diet (5%), HS: high-starch diet (19%) and SB: high-starch diet with BAs (350 mg/kg diet). An 8-week feeding trial was conducted in largemouth bass of initial weight 23.69 ± 0.13 g. The results indicated that the weight gain (WG) and protein efficiency ratio (PER) of fish fed LS and SB were significantly higher than HS treatment. The superoxide dismutase (SOD) and catalase (CAT) activities of SB group were significantly increased, while malondialdehyde (MDA) content significantly reduced in liver compared with HS group. The activities of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and glucose contents in plasma of SB group were significantly lower than HS treatment, whereas the content of triglyceride (TG) and total cholesterol (TC) in plasma were significantly higher than HS treatment. Additionally, the plasma immunoglobulin count, lysozyme activity and the blood leukocyte count (WBC) in SB group were significantly higher than HS group. The results of paraffin section of liver showed the histopathological alterations were significantly reduced in the SB group compared to HS group. All in all, this study revealed that bile acids supplement could significantly improve growth performance, enhance liver function and immune ability, and alleviate stress responses of M. salmoides fed high-starch diet.

High dietary lipid level alters the growth, hepatic metabolism enzyme, and anti-oxidative capacity in juvenile largemouth bass Micropterus salmoides.

The present study was conducted to investigate the effects of high dietary lipid levels on growth, metabolism, antioxidant capacity, and immune responses of largemouth bass. Fish (initial body weight 13.38 ± 0.11 g) were fed three isonitrogenous semi-purified diets containing 5%, 10%, and 20% lipid, respectively. The results indicated that fish fed 10% lipid diet showed significantly better final body weight, specific growth rate (SGR), protein efficiency ratio (PER), and feed conversion ratio (FCR) compared with that fed 5% lipid diet. Meanwhile, fish fed 20% lipid diet had a significantly higher viscera ratio (VR), hepatosomatic index (HSI), intraperitoneal fat ratio (IPF), and liver lipid content than those fed the other diets. Higher alanine aminotransferase (ALT) and aspartate transaminase (AST) activities, total cholesterol (TC), triglyceride (TG), free fatty acids (FFA), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) contents, and LDL-C/HDL-C value in plasma were recorded in fish fed 20% lipid diet, while higher insulin contents were obtained in fish fed 5% lipid diet. In addition, the highest carnitine palmitoyltransferase I (CPT1), AMP-activated protein kinase (AMPK), fructose-1,6-bisphosphatase (FBPase), and phosphoenolpyruvate carboxykinase (PEPCK) activities in the liver were also observed in fish fed 20% lipid diet. However, fish fed 20% lipid diet had a significantly lower superoxide dismutase (SOD) and catalase (CAT) activities and higher MDA contents in liver than those fed the other diets. The higher nitric oxide (NO) contents and inducible nitric oxide synthase (iNOS) activity in liver were recorded in fish fed 10% lipid diet. Moreover, the alkaline phosphatase (ALP), inducible nitric oxide synthase (iNOS) and lysozyme activities, and nitric oxide (NO) contents in plasma were higher in fish fed the 10% diets than the other groups. In conclusion, high dietary lipid levels could suppress growth performance and liver anti-oxidative capacity, and reduce immune responses of largemouth bass.

Linseed oil can decrease liver fat deposition and improve antioxidant ability of juvenile largemouth bass, Micropterus salmoides.

A feeding trial was conducted to evaluate the effect of linseed oil (LO) on growth, plasma biochemistry, hepatic metabolism enzymes, and antioxidant capacity of juvenile largemouth bass, Micropterus salmoides. Four isonitrogenous (crude protein, 45%) and isoenergetic (gross energy, 18 MJ/kg) diets were formulated by replacing 0 (the control), 33.3%, 66.7%, and 100% of fish oil with linseed oil. Each diet was fed to three replicate groups of fish (initial body weight, 22.02 ± 0.61 g) for 8 weeks. The results indicated that fish fed diet with 100% LO substitution level had lower weight gain (WG), specific growth rate (SGR), and protein efficiency ratio (PER) than the other groups (P < 0.05), while feed conversion ratio (FCR) was higher compared to the other groups (P < 0.05). Feed intake (FI) and hepatosomatic index (HSI) of 66.7% LO substitution level were significantly lower than the control groups (P < 0.05). Glycogen, lipid, and non-esterified fatty acid content in the liver decreased significantly with increasing dietary LO levels (P < 0.05). Moreover, the replacement of fish oil (FO) with LO could significantly reduce the content of triglyceride (TG) and total cholesterol (TC) and the activity of alanine amiotransferase (ALT) in plasma of M. salmoides (P < 0.05). There were significant differences in hepatic metabolism enzymes in fish fed diets with different dietary LO levels. Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor (PPAR-α) activities in liver significantly increased with increasing dietary LO level (P < 0.05). In addition, phosphoenolpyruvate carboxykinase (PEPCK) and fructose-1,6-bisphosphatase (FBPase) activities in the liver significantly increased with decreasing dietary LO level (P < 0.05). Both the lowest superoxide dismutase (SOD) and catalase (CAT) activities in the liver were recorded in the control group (P < 0.05). Moreover, nitric oxide content, glutathione peroxidase (GPx), and inducible nitric oxide synthase (iNOS) activities in the liver significantly increased with increasing dietary LO level, while malondialdehyde (MDA) content significantly reduced. These findings demonstrated that LO can improve liver function and antioxidant ability of M. salmoides. In addition, replacing partial FO with LO cannot affect growth performance, but all substitutions inhibit growth performance of M. salmoides.

Molecular and metabolic adaption of glucose metabolism in the red and white muscle of the omnivorous GIFT tilapia Oreochromis niloticus to a glucose load.

In this experiment, Genetically improved farmed Nile tilapia Oreochromis niloticus were intraperitoneally injected with 1 g glucose/kg of body weight or saline. Red and white muscle tissues were collected at 0, 1, 2, 4, 6 and 12 h after the glucose tolerance test (GTT) or saline injection, and the time course of changes in molecular and metabolic adaption of glucose metabolism of these two tissues were evaluated. The results showed that the expression of insulin-responsive glucose transporter 4 (glut4) was up-regulated at 4 h after the GTT in the red muscle, implying an increase of glucose uptake. However, the expression of glut4 in the white muscle did not change with glucose load. The glycolysis of red muscle in tilapia was stimulated during 2-4 h after the GTT, as the expression of hexokinase 1b (hk1b), hk2, phosphofructokinase muscle type a (pfkma) and pfkmb and the activity of HK and PFK increased. By contrast, only the expression of hk1b was up-regulated at 6 h after the GTT in the white muscle. The mRNA level of glycogen synthase 1 (gys1) and glycogen content increased at 2 and 6 h, respectively after the GTT in the red muscle, suggesting that glucose storage was provoked. However, glycogen content in the white muscle was not impacted by GTT. Lipogenesis was stimulated in the red muscle as reflected by up-regulated expression of acetyl-CoA carboxylase α (accα) (during 2-4 h) and accß (during 4-12 h) with GTT. In the white muscle, however, the expression of accα was not changed, and mRNA level of accß was not up-regulated until 6 h after the GTT. Taken together, it was concluded that the glycolytic and glycogen synthesis mechanisms in the red muscle were highly regulated by an acute glucose load while those in the white muscle were less responsive to this stimulus.

Effect of high dietary starch levels on growth, hepatic glucose metabolism, oxidative status and immune response of juvenile largemouth bass, Micropterus salmoides.

An experimental trial was conducted to investigate the effects of high dietary starch levels on growth, hepatic glucose metabolism enzyme, antioxidant capacity and immune responses of largemouth bass, Micropterus salmoides. Fish (initial body weight: 16.9 ±â€¯0.24 g) were fed three isonitrogenous and isoenergetic semi-purified diets containing 5%, 10% and 20% wheat starch, respectively. The results indicated that fish fed 5% and 10% starch diets showed significantly better weight gain, specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) compared with that fed 20% starch diet. Meanwhile, fish fed 20% starch diet had a significantly higher hepatic glycogen and muscle glycogen contents than those fed the other diets. The alanine amiotransferase (ALT) and aspartate transaminase (AST) activities, glucose and insulin contents in plasma increased significantly with dietary starch levels, whereas triglyceride content showed the opposite trend. In addition, the highest glucokinase (GK), pyruvate kinase (PK) and phosphofructokinase (PFK) activities in liver were also observed in fish fed 20% starch diet. However, both fructose-1,6-bisphosphatase (FBPase) and pyruvate carboxylase (PC) activities in liver decreased significantly as dietary starch levels increased. Moreover, the lower superoxide dismutase (SOD) and catalase (CAT), the higher malondialdehyde (MDA) contents in liver were observed in fish fed 20% starch diets. Compared to the 5% and 10% starch, the 20% starch could enhance the content of plasma nitric oxide (NO) and the activities of inducible nitric oxide synthase (iNOS) and alkaline phosphatase (ALP). Results demonstrate that the starch levels may affect growth performance and metabolic changes, which suggest that high-starch diets were inefficiently used as an energy source by M. salmoides juveniles. Excessive dietary starch contents could result in oxidative stress, suppress innate immunity, and thus affect the health status of M. salmoides.

Effect of dietary phospholipid levels on growth, lipid metabolism, and antioxidative status of juvenile hybrid snakehead (Channa argus×Channa maculata).

The study was conducted to evaluate the effect of dietary phospholipids (PLs) on growth, lipid metabolism, and antioxidative status of hybrid snakehead (Channa argus × Channa maculata). Five isonitrogenous and isolipidic diets with graded levels of PLs (8.5, 19.3, 30.7, 41.5, and 50.8 g kg-1) were fed to triplicate groups of juveniles (initial body weight 12.6 ± 0.23 g) for 8 weeks. Results showed that dietary PL supplementation significantly improved growth of juveniles. The final body weight (FBW) and specific growth rate (SGR) significantly increased with dietary PLs increasing from 8.5 to 41.5 g kg-1 (P < 0.05). Fish fed with the diet containing 8.5 g kg-1 PLs showed higher feed conversion ratio (FCR) compared to the other treatments (P < 0.05). Survival rate (SR) was not affected by dietary PL levels (P > 0.05). Liver lipid contents, serum triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) contents significantly decreased with the increasing levels of dietary PLs (P < 0.05). However, serum total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) contents and HDL-C/TC and HDL-C/LDL-C value significantly increased with increasing dietary PL levels (P < 0.05). The catalase (CAT), superoxide dismutase (SOD), and carnitine palmitoyl transferase I (CPT-1) activities in the liver significantly increased with incremental dietary PL level (P < 0.05), while the liver malondialdehyde (MDA) contents and fatty acid synthase (FAS) activity significantly reduced (P < 0.05). No significant difference was observed in the glutathione peroxidase (GPx) activity among dietary treatments (P > 0.05).These results confirmed that dietary PL supplementation has beneficial effects on growth performance and antioxidant capacity of juvenile hybrid snakehead. Dietary PLs might reduce lipid deposition in the liver of juvenile hybrid snakehead.

Effects of Astragalus polysaccharides (APS) and chitooligosaccharides (COS) on growth, immune response and disease resistance of juvenile largemouth bass, Micropterus salmoides.

The effects of oral administration of Astragalus polysaccharides (APS) and chitooligosaccharides (COS), single or combined, on the growth performance, immunity and disease resistance of M. salmoides were investigated. Largemouth bass juvenile were divided into 4 groups and each group was fed with diets supplemented with or without immunostimulant for 8 weeks. After 8 weeks of feeding trial, five fish per tank were sampled for immunity determination, ten fish per tank were challenged by A. hydrophila. The results showed that the largemouth bass fed with two immunostimulants alone or in combination significantly enhanced the final weight and specific growth rate (SGR), decreased feed conversion ratio (FCR) (P < 0.05). However, there were no significant differences (P < 0.05) in specific growth rate (SGR) between dietary COS and dietary APS. In addition, both COS and APS upregulated respiratory burst activity (RBA), phagocytic activity (PA), lysozyme activity and superoxide dismutase (SOD) activity. Meanwhile, COS also exhibited a increase in total leukocyte count, nitric oxide (NO) content and inducible nitric oxide synthase (iNOS) activity compared to the control. When challenged with A. hydrophila, the mortality of groups fed with COS and/or APS was lower than the control (P < 0.05). Under the experimental conditions, dietary APS and COS had a synergistic effect on lysozme activity, iNOS activity, NO content and disease resistance of fish (P < 0.05).

Simultaneous stimulation of glycolysis and gluconeogenesis by feeding in the anterior intestine of the omnivorous GIFT tilapia, Oreochromis niloticus.

The present study was performed to investigate the roles of anterior intestine in the postprandial glucose homeostasis of the omnivorous Genetically Improved Farmed Tilapia (GIFT). Sub-adult fish (about 173 g) were sampled at 0, 1, 3, 8 and 24 h post feeding (HPF) after 36 h of food deprivation, and the time course of changes in intestinal glucose transport, glycolysis, glycogenesis and gluconeogenesis at the transcription and enzyme activity level, as well as plasma glucose contents, were analyzed. Compared with 0 HPF (fasting for 36 h), the mRNA levels of both ATP-dependent sodium/glucose cotransporter 1 and facilitated glucose transporter 2 increased during 1-3 HPF, decreased at 8 HPF and then leveled off. These results indicated that intestinal uptake of glucose and its transport across the intestine to blood mainly occurred during 1-3 HPF, which subsequently resulted in the increase of plasma glucose level at the same time. Intestinal glycolysis was stimulated during 1-3 HPF, while glucose storage as glycogen was induced during 3-8 HPF. Unexpectedly, intestinal gluconeogenesis (IGNG) was also strongly induced during 1-3 HPF at the state of nutrient assimilation. The mRNA abundance and enzyme activities of glutamic-pyruvic and glutamic-oxaloacetic transaminases increased during 1-3 HPF, suggesting that the precursors of IGNG might originate from some amino acids. Taken together, it was concluded that the anterior intestine played an important role in the regulation of postprandial glucose homeostasis in omnivorous tilapia, as it represented significant glycolytic potential and glucose storage. It was interesting that postprandial IGNG was stimulated by feeding temporarily, and its biological significance remains to be elucidated in fish.

Dietary administration of chitooligosaccharides to enhance growth, innate immune response and disease resistance of Trachinotus ovatus.

The present study was conducted to investigate the effects of dietary chitooligosaccharides (COS) supplementation on the innate immune response and protection against Vibrio harveyi infection in Trachinotus ovatus. A basal diet was supplemented with 0.0 (control), 2.0, 4.0 and 6.0 g COS kg(-1) to formulate four experimental diets. Each diet was randomly allocated to triplicate groups of fish in floating sea cages (1.5 × 1.0 × 2.0 m), and each cage was stocked with 80 fish (initial average weight 10.8 ± 0.05 g). After 8 weeks of feeding trial, Both the final weight and specific growth rate (SGR) significantly increased with increasing dietary COS levels up to 4.0 g kg(-1), whereas there were no significant differences for COS levels from 4.0 to 6.0 g kg(-1). A decreased feed conversion ratio (FCR) was observed with increasing dietary COS levels. The total leukocyte counts (WBC), differential leukocyte counts, respiratory burst activity, lysozyme and superoxide dismutase (SOD) activity were significantly increased with the increased levels of dietary COS (P < 0.05), and reached a maximum at level of 4.0 g kg(-1) COS. There were no significant differences in those immunological parameters between 4.0 and 6.0 g kg(-1) COS. Moreover, the dietary COS supplementation groups also exhibited a decrease in the cumulative symptom rates compared to the controls when challenged with V. harveyi. These results indicated that dietary intake containing COS could enhance the immune responses of fish and improve its resistance to infection by V. harveyi. Especially supplementation with 4.0 g kg(-1) COS to the fish for 56 days showed considerable improvement in the growth, survival and immune response of the fish.

Effects of dietary ß-1,3-glucan, chitosan or raffinose on the growth, innate immunity and resistance of koi (Cyprinus carpio koi).

This study was performed to determine the efficacy of three immunomodulators viz., ß-1,3 glucan, chitosan and raffinose on the innate immune response of koi, Cyprinus carpio koi. Kois were divided into 4 groups and each group was fed with diets supplemented with or without immunostimulant for 56 days. Total leukocyte counts (WBC), the non-specific humoral (lysozyme, alternative complement pathway and superoxide dismutase) and cellular (phagocytic capacity and respiratory burst activity) responses were determined and compared with controls (no supplement) after 7, 14, 21 and 56 days of feeding. The results of 8 weeks feeding trial showed that ß-1,3 glucan supplementation significantly enhanced koi growth, whereas other immunostimulants did not. Variation in the levels of responses was evident among different supplements. Compared with chitosan or raffinose, ß-1,3 glucan could maintain the immunity of kois at a higher level during the experimental period. However, continuously applying ß-1,3 glucan, chitosan or raffinose into the diet caused immunity fatigue in koi. No significant change in alternative complement pathway (ACP) activity was observed for any of the three supplements over the four different periods. After feeding for 14 days, the total leukocyte count (WBC), respiratory burst activity and superoxide dismutase (SOD) activity of the kois fed with chitosan or raffinose continuously remained relatively unchanged, subsequently decreased on the 56th day, but SOD did not. Meanwhile, lysozyme activity was no longer significantly higher on the 7th day, and for phagocytic capacity on the 14th day. After 56 days, these three immunostimulants groups also exhibited a decrease in the cumulative symptom rates compared to the controls when challenged with Aeromonas veronii. These results indicated that dietary intake containing immunostimulants could enhance the immune responses of koi and improve its resistance to infection by A.veronii. Especially supplementation with ß-1,3 glucan to the kois for 56 days showed considerable improvement in the growth, survival and immune response of the kois.