Modulation of appetite,lipid and glucose metabolism of juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) by different dietary protein levels

This study was undertaken to explore the systemic metabolic strategies of juvenile grass carp (Ctenopharyngodon idellus) to maintain growth when fed with different dietary protein levels. The optimal growth group and two growing discomfort groups were selected through the basic data, to explain the growth difference from appetite regulation and lipid and glucose metabolism perspective. Three experimental diets were formulated with three dietary protein levels at 200.3, 296.1 and 442.9 g kg^?1, named P1, P2 and P3, respectively. Juvenile grass carp (initial body weight 12.28 ± 0.14 g) were fed with three diets with 3 replications per dietary treatment in an indoor recirculation system for an 8-week feeding trial. Fish fed with diet P2 dietary group showed significantly higher WG, SGR, FI and PER than other groups. Compared with other groups, mRNA expressions of NPY, Y8a and Y8b in fish fed with P2 significantly down-regulated, while the expressions of CCK and CART in fish fed with P3 significantly down-regulated (P < 0.05). With increasing dietary protein levels, G6Pase, GK, PK and PEPCK were all significantly inhibited (P < 0.05). For lipid metabolism, the mRNA expression of ACC in P1 dietary group was significantly higher than P3 dietary group; besides, LPL expression in P3 group was significantly higher than other two groups (P < 0.05). PPARα expression in P2 was significantly lower than other groups (P < 0.05). These results suggested that grass carp fed with P2 (296.1 g kg^?1 protein level) showed highest weight gain, contributed to more balanced nutrient metabolism and appetite regulation. Too high dietary protein (442.9 g kg^?1) should be avoided because it induced lowest PER, body lipid and liver lipid, and inhibited glucose and lipid metabolism in juvenile grass carp.     

Fat deposition pattern and mechanism in response to dietary lipid levels in grass carp, <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>

This study aimed to evaluate the fat deposition pattern and lipid metabolic strategies of grass carp in response to dietary lipid levels. Five isonitrogenous diets (260 g kg^?1 crude protein) containing five dietary lipid levels (0, 20, 40, 60, 80 g kg^?1) were fed to quadruplicate groups of 15 fish with initial weight 200 g, for 8 weeks. The best growth performance and feed utilization was observed in fish fed with lipid level at 40 g kg^?1. MFI and adipose tissue lipid content increased with increasing dietary lipid level up to 40 g kg^?1, and higher lipid level in diet made no sense. Fish adapted to high lipid intake through integrated regulating mechanisms in several related tissues to maintain lipid homeostasis. In the present study, grass carp firstly increased PPARγ and CPT1 expressions in adipose tissue to elevate adipocyte differentiation and lipolysis to adapt to high lipid intake above 40 g kg^?1. In liver, fish elevated hepatic lipid uptake but depressed biosynthesis of hepatic FAs, resulted in no difference in HSI and liver lipid content among the groups. Only in muscle, fish showed a significant fat deposition when the lipid intake above 40 g kg^?1. The excess lipid, derived from enhanced serum TC and TG contents, was more likely to induce deposition in muscle rather than lipid uptake by adipose tissue in grass carp fed with high dietary lipid, indicating the muscle of grass carp might be the main responding organ to high lipid intake.     

Dietary docosahexaenoic acid decreased lipid accumulation via inducing adipocytes apoptosis of grass carp, <Emphasis Type="Italic">Ctenopharygodon idella</Emphasis>

The purpose of this study was to explore the mechanism of by which docosahexaenoic acid (DHA) inhibit the accumulation of adipose tissue lipid in grass carp (Ctenopharyngodon idella). We therefore designed two semi-purified diets, namely DHA-free (control) and DHA-supplemented, and fed them to grass carp (22.19 ± 1.76 g) for 3 and 6 weeks. DHA supplementation led to a significantly lower intraperitoneal fat index (IPFI) than that in the control group by reducing the number of adipocytes but significantly higher adipocyte size (P < 0.05). In the intraperitoneal adipose tissue, the DHA-fed group showed significantly higher peroxisome proliferator-activated receptor (PPAR)γ, CCAAT enhancer-binding protein (C/EBP)α, and sterol regulatory element-binding protein (SREBP)1c mRNA expression levels at both 3 and 6 weeks (P < 0.05). However, the ratio of the expression levels of B cell leukemia 2 (Bcl-2) and Bcl-2-associated X protein (Bax) was significantly lower in the DHA-fed group than in the control group (P < 0.05), and the protein expression levels of the apoptosis-related proteins caspase 3, caspase 8, and caspase 9 were also significantly higher (P < 0.05). Overall, although DHA promotes lipid synthesis, it is more likely that DHA could suppress the lipid accumulation in adipocytes of grass carp by inducing adipocyte apoptosis.     

Stimulation of glycerol kinase in grass carp preadipocytes by EPA

This study was conducted to assess the effect of eicosapentaenoic acid (EPA) on grass carp preadipocyte glycerol kinase (GyK) expression, as well as to explore the mechanism. Here, we cloned partial sequence of grass carp GyK gene and analyzed its tissue distribution. The result showed that GyK gene expressed most in the liver, followed by adipose tissue and the kidney. Besides, 400 μM oleic acid (18:1n-9, OA) was used to establish a hypertrophic preadipocyte model. GyK gene expression and enzyme activity were significantly enhanced after model cells were treated with 100 μM eicosapentaenoic acid (20:5n-3, EPA) for 6, 12, and 24 h. Meanwhile, peroxisome proliferative-activated receptor (PPAR)γ, adipose triglyceride lipase (ATGL), and the two isoforms of grass carp HSL gene were first identified by Sun et al (2016), and they defined the two isoforms as HSLa and HSLb. Therefore, maybe HSLa and HSLb are appropriate.. The content of triglyceride was dramatically increased by EPA treatment for 24 h. Further, a competitive ATGL antagonist, HY-15859, attenuated the increase in GyK induced by EPA at 12 h. Surprisingly, the enhanced lipolysis and PPARγ gene expression induced by serum deprivation were paralleled by an increase in GyK gene expression, whereas a stabilization in GyK enzyme activity. Other fatty acids, including docosahexaenoic acid, alpha-linolenic acid, linoleic acid, and OA also promoted GyK gene expression. Moreover, an irreversible PPARγ antagonist, GW9662, was used to investigate the role of PPARγ in GyK induction. Data showed that GW9662 abolished the induction of GyK by EPA at 12 h. Together, these data suggested that EPA elevated grass carp preadipocytes GyK expression. ATGL and PPARγ contributed to the induction of GyK. PPARγ may be a key regulator in response to GyK expression induced by EPA.     

Effect of waterborne zinc exposure on lipid deposition and metabolism in hepatopancreas and muscle of grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

The aim of the present study was to explore the effect of waterborne zinc (control, 0.85, 2.20, 3.10 mg/l, respectively) exposure on lipid deposition and metabolism in the hepatopancreas and muscle of grass carp Ctenopharyngodon idella. The lipid content, Zn accumulation, and the activities and expression levels of several enzymes involved in lipid metabolism were determined in hepatopancreas and muscle. Waterborne Zn exposure reduced growth performance and increased Zn accumulation in both tested tissues. In hepatopancreas, Zn exposure increased lipid content, the activities of lipogenic enzymes, such as 6PGD, G6PD, ME, ICDH and FAS, as well as the mRNA expression level of G6PD, 6PGD, ICDH, FAS and SREBP-1. But the activity of CPT I and the mRNA expression of HSL, CPT Iα1a, CPT Iα2a and PPARα were down-regulated by Zn exposure. In contrast, in muscle, waterborne Zn exposure decreased lipid deposition, activities of 6GPD, ICDH and ME, as well as the mRNA expression level of G6PD, ICDH, ME, FAS and SREBP-1. However, the activity of CPT I as well as the mRNA expression level of PPARα, HSL, CPT Iα2a, CPT Iα1b and CPT Iβ were up-regulated by Zn exposure. Our results indicate that waterborne Zn increases lipid content by up-regulating lipogenesis and down-regulating lipolysis in hepatopancreas. But, in muscle, waterborne Zn reduces lipid accumulation by up-regulating lipolysis and down-regulating lipogenesis. Differential patterns of lipid deposition, enzymatic activities and genes’ expression indicate the tissue-specific regulatory mechanism in fish.     

Cloning and characterization of <Emphasis Type="Italic">Bax1</Emphasis> and <Emphasis Type="Italic">Bax2</Emphasis> genes of <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> and evaluation of transcript expression in response to grass carp reovirus infection

Multidomain proapoptotic Bcl-2-associated X (Bax) protein is an essential effector responsible for mitochondrial outer membrane permeabilization, resulting in cell death via apoptosis. In this study, two Bax genes of grass carp (Ctenopharyngodon idellus), designated as CiBax1 and CiBax2, were isolated and analyzed. The obtained CiBax1 cDNA is 2058 bp long, with a 579 bp open reading frame (ORF) coding a protein of 192 amino acid residues. The full-length cDNA of CiBax2 is 1161 bp, with a 618 bp ORF coding 205 amino acids. Both CiBax1 and CiBax2 are typical members of Bcl-2 family containing conserved Bcl and C-terminal domains, and they share conserved synteny with zebrafish Bax genes despite the grass carp Bax mapping to different linkage groups. Phylogenetic analysis showed that CiBax1 was clustered with Bax from most teleost fish, and CiBax2 was close to Bax2 from teleost fish but far separated from that of Salmo salar. Quantitative real-time PCR analysis revealed broad expression of CiBax1 and CiBax2 in tissues from healthy grass carp, but the relative expression level differed. The mRNA expression of CiBax1 and CiBax2 was both upregulated significantly and peaked in all examined tissues at days 5 or 6 post-infection with grass carp reovirus. Subcellular localization indicated that CiBax1 protein was localized in both nucleus and cytosol, while CiBax2 protein only in cytosol. Moreover, CiBax2, but not CiBax1 was colocalized with mitochondrion under normal condition. Taken together, the findings would be helpful for further understanding of the function of Bax in teleost fish.     

Role of cyclooxygenase-mediated metabolites in lipid metabolism and expression of some immune-related genes in juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) fed arachidonic acid

Cyclooxygenase (COX) catalyzes the conversion of arachidonic acid (ARA) to prostaglandins, and COX-mediated metabolites play important roles in the regulation of lipid metabolism and immunity in mammals. However, such roles of COX in fish remain largely unknown. In this study, we designed three semi-purified diets, namely ARA-free (control), ARA, and ARA + acetylsalicylic acid (ASA; a COX inhibitor), and used them to feed grass carp (27.65 ± 3.05 g) for 8 weeks. The results showed that dietary ARA significantly increased the amount of ARA in the hepatopancreas, muscle, and kidney (P < 0.05), whereas this increase was reduced by dietary ASA. The hepatopancreatic prostaglandin E₂ content increased in the ARA group, and this increase was inhibited by ASA (P < 0.05). ARA decreased the lipid content in the hepatopancreas, whereas ASA recovered lipid content to a significant level (P < 0.05). ARA significantly decreased the messenger RNA (mRNA) expression levels of fatty acid synthase and stearoyl-CoA desaturase in the hepatopancreas (P < 0.05). However, ASA did not rescue the mRNA expression of these genes (P > 0.05). Interestingly, ARA significantly enhanced the level of peroxisome proliferator-activated receptor α gene expression, and this increase was attenuated by ASA (P < 0.05). Finally, ARA significantly enhanced the mRNA expression of myeloid differentiation factor 88 (MyD88) in the kidney, and ASA attenuated the expression of toll-like receptor 22 and MyD88 (P < 0.05). In conclusion, our findings suggest that COX metabolites play important roles in the inhibition of lipid accumulation in the hepatopancreas of grass carp fed with ARA and that regulation of gene expression promotes lipid catabolism rather than lipogenic activities. Additionally, these eicosanoids might participate in the upregulation of immunity-related genes in the kidney.     

A study of the damage of the intestinal mucosa barrier structure and function of <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis> with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

The aim of this study is to explore the effect of Aeromonas hydrophila on the intestinal mucosal barrier structure and intestinal permeability in grass carp (Ctenopharyngodon idella). Histopathological examinations showed that A. hydrophila induced severe intestinal lesions, including inflammatory cell infiltration and intestinal villus fusion and swelling. Messenger RNA (mRNA) expression of the inflammatory cytokines TNF-α, IL-1β, IL-8, IL-10 and MyD88 was significantly increased after infection with A. hydrophila. The permeability of intestinal mucosa was determined using Evans blue (EB) and d-lactic acid. The results indicated that the levels of EB and serum d-lactic acid were significantly increased after infection with A. hydrophila (p < 0.05). Our results also indicated that the intestinal mucosal barrier injury induced by A. hydrophila infection was closely associated with the expression of the tight junction (TJ) protein zonula occludens-1 (ZO-1), occludin, claudin b and claudin c as well as the activity of Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase. Lower mRNA levels of occludin and lower Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activity in the intestines were observed after challenge. ZO-1 and claudin c were significantly increased 24 h after infection with A. hydrophila. The most interesting finding was that claudin b also significantly increased 24 h after challenge and then decreased to lower levels at 72, 120 and 168 h post-infection compared to the PBS-treated control group. The results demonstrated that grass carp infection with A. hydrophila induced intestinal inflammation and impaired the structure and function of the intestinal mucosal barrier.     

Corn gluten meal in pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> diets: effects on growth,haematology, and meat quality

Corn gluten meal (CGM) has a high protein content and absence of antinutrients. However, it has high levels of carotenoids that can cause a yellowing of fish fillets and impair further commercialisation. Graded levels of CGM were incorporated in pacu diets to replace soybean meal (SM) protein. The experimental diets were formulated to be isonitrogenous (220 g kg^?1 digestible protein) and isoenergetic (13.4 MJ kg^?1 digestible energy) with 0, 25, 50, 75, and 100 % replacement of SM protein by CGM protein over a 60-day period. One hundred fish (32.79 ± 3.43 g) were randomly distributed in 20 experimental cages (70 L) placed within five (1000 L) aquaria with continuous water renewal. The experimental design was completely randomised with five treatments and four replicates. A quadratic effect (P < 0.05) was observed for fillet and carcass yields, feed conversion ratio, weight gain, and specific growth rate, with the optimum values for replacement of SM protein by CGM protein estimated as 21.95, 29.13, 30.94, 37.71, and 38.75 %, respectively. There were no differences (P > 0.05) for blood parameters, proximal composition, water-holding capacity, pH, lipid oxidation, and a* value of fillets. The L* and b* values of fillets showed differences (P < 0.05). Replacement between 21.95 and 38.75 % of SM protein by CGM protein (5.37 and 9.48 % of inclusion in the diet) improves the growth performance and body yield of pacu juveniles without altering blood parameters or affecting the fillet quality.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Nitrogen removal characteristics of <Emphasis Type="Italic">Pseudomonas stutzeri</Emphasis> F11 and its application in grass carp culture

This study was designed to investigate the denitrification characteristics of Pseudomonas stutzeri F11 under different environmental conditions and to evaluate the effect of these characteristics on nitrogen removal and the water microbiome in an experimental grass carp aquaculture system. The results showed that the optimal growth conditions of strain F11 were (1) ammonia-nitrogen (N), nitrite-N, or nitrate-N as sole N source; (2) initial ammonia-N concentration of 10–30 mg N/l; (3), initial nitrite-N concentration of 200 mg N/l; (4) sodium citrate as carbon source; (5) rotation (r) speed of 200 r/min; a C:N ratio of between 2 and 10; (6) culture at 32–37 °C. The addition of P. stutzeri F11 to the experimental grass carp aquaculture system reduced the levels of ammonia-N, nitrite-N, and total N in the water over an extended range, but had no effect on nitrate-N level. Results of the 454 pyrosequencing analysis indicated that the structure of the microbial community in the aquaculture water changed significantly after the addition of P. stutzeri F11 preparations. The addition of P. stutzeri F11 to the aquaculture system also altered the microbiome metabolism in the water, especially the bacteria involved in nitrogen metabolism. These results suggest that the addition of P. stutzeri F11 to an experimental grass carp aquaculture system decrease nitrogen levels and alter the microbial community structure of the water; as such, this bacterial strain could be a potential candidate for the regulation of water quality in aquaculture systems.     

Respiratory response of grass carp <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> 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 (f_R), oxygen consumption rate (VO₂), respiratory stroke volume (V_S.R), gill ventilation (V_G), and V_G/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 f_R, V_S.R, V_G, and V_G/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 f_R, V_S.R, V_G, and V_G/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 (C_c) 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.     

Establishment and characterization of a heart-derived cell line from goldfish (<Emphasis Type="Italic">Carassius auratus</Emphasis>)

The goldfish Carassius auratus, a freshwater fish in the family Cyprinidae, was one of the earliest fish to be domesticated for ornamental purposes. A cell line was established from goldfish heart (GH) tissue to create a biological monitoring tool for viral diseases. The GH cell line was optimally maintained at 25 °C in M199 medium supplemented with 10–20% fetal bovine serum. A chromosomal analysis indicated that the cell line remained diploid, with a mean chromosomal count of 100. In viral inoculation assays, significant cytopathic effects (CPEs) were caused by epizootic hematopoietic necrosis virus (EHNV), Andrias davidianus iridovirus (ADIV), and Bohle iridovirus (BIV) infections in the fish cells and the viral titers (average value) of EHNV, ADIV, and BIV in GH cells reached 10^5.0, 10^4.5, and 10^5.0 TCID₅₀/0.1 mL, respectively, within 7 days. However, no CPE was observed in the cells infected with viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), spring viremia of carp virus (SVCV), infectious pancreatic necrosis virus (IPNV), channel catfish virus (CCV), or grass carp reovirus (GCRV). These results suggest that the GH cell line is a valuable tool for studying viral pathogenesis.     

Effects of stocking density on lipid deposition and expression of lipid-related genes in Amur sturgeon (<Emphasis Type="Italic">Acipenser schrenckii</Emphasis>)

To investigate the correlation between lipid deposition variation and stocking density in Amur sturgeon (Acipenser schrenckii) and the possible physiological mechanism, fish were conducted in different stocking densities (LSD 5.5 kg/m³, MSD 8.0 kg/m³, and HSD 11.0 kg/m³) for 70 days and then the growth index, lipid content, lipase activities, and the mRNA expressions of lipid-related genes were examined. Results showed that fish subjected to higher stocking density presented lower final body weights (FBW), specific growth ratio (SGR), and gonad adipose tissue index (GAI) (P < 0.05). Lower lipid content was observed in the liver, gonad adipose tissue and muscle in sturgeons held in HSD group (P < 0.05). The serum concentrations of triglyceride (TG), total cholesterol (TC), and high-density lipoprotein cholesterol (HDL-C) decreased significantly with increasing stocking density, while no significant change was observed for low-density lipoprotein cholesterol (LDL-C). Furthermore, the cDNAs encoding lipoprotein lipase (LPL) and hepatic lipase (HL) were isolated in Amur sturgeon, respectively. The full-length LPL cDNA was composed of 1757 bp with an open reading frame of 501 amino acids, while the complete nucleotide sequences of HL covered 1747 bp encoding 499 amino acids. In the liver, the activities and mRNA levels of LPL were markedly lower in HSD group, which were consistent with the variation tendency of HL. Fish reared in HSD group also presented lower levels of activities and mRNA expression of LPL in the muscle and gonad. Moreover, the expressions of peroxisome proliferator-activated receptor α (PPARα) in both the liver and skeletal muscle were significantly upregulated in HSD group. Overall, the results indicated that high stocking density negatively affects growth performance and lipid deposition of Amur sturgeon to a certain extent. The downregulation of LPL and HL and the upregulation of PPARα may be responsible for the lower lipid distribution of Amur sturgeon in higher stocking density.     

Protein‐sparing capability of dietary lipid in herbivorous and omnivorous freshwater finfish: a comparative case study on grass carp (Ctenopharyngodon idella) and tilapia (Oreochromis niloticus × O. aureus)

Two, 8‐week feeding trials were conducted to compare protein‐sparing capability of dietary lipid in herbivorous grass carp (Ctenopharyngodon idella) and omnivorous tilapia (Oreochomis niloticus × O. aureus). Utilizing a 2 × 3 factorial design, experimental diets containing two levels of crude protein (380 and 250 g kg⁻¹) and three levels of lipid (0, 40 and 100 g kg⁻¹) were formulated for use in both feeding trials. Growth performances showed better response of both fish fed 380 g kg⁻¹ protein diet than those fed 250 g kg⁻¹ protein diet. Despite the dietary protein level, weight gain (WG), specific growth ratio (SGR), feed conversion ratio (FCR) and protein efficiency ratio were much higher (P < 0.05) for grass carp fed 40 g kg⁻¹ lipid diet than those fed 100 g kg⁻¹ lipid diet; however, there were no significant differences in tilapia fed the two diets. The feed intake of grass carp fed lipid‐free diet was the lowest, but it tended to decrease with increase in dietary lipids in tilapia. Lipid retention (LR) was negatively correlated with dietary lipid concentration of both fish. Viscerosomatic index (VSI), hepatosomatic index (HSI), intraperitoneal fat ratio (IPF) and whole‐body and liver lipid content positively correlated with dietary lipid concentration of both fish. Plasma parameters and liver enzymes activities were also positively correlated with dietary lipid concentration of both fish. Liver lipid contents were higher and enzymes activities were lower in grass carp when compared with tilapia. These data suggested that there was no evidence of a protein‐sparing effect of dietary lipids in grass carp. Tilapia has relatively higher capacity to endure high dietary lipid level compared to grass carp.     

注射硫代乙酰胺并饲喂酵母培养物、姜黄素和水飞蓟素对草鱼脂代谢相关基因表达丰度的影响

为探讨硫代乙酰胺(TAA)诱导草鱼肝胰脏损伤与损伤修复的作用机制, 试验以腹腔注射硫代乙酰胺诱导肝胰脏损伤实验模型草鱼为对象, 分别饲喂含有酵母培养物dv、姜黄素和水飞蓟素的饲料70 d后, 采集草鱼肝胰脏样品, 采用实时定量反转录聚合酶链式反应(qPCR)方法, 检测了草鱼肝胰脏脂肪酸合成酶(FAS)、过氧化物酶增殖体激活受体γ辅助激活因子(PGC1-α)、硬脂酰辅酶A去饱和酶1(SCD1)、解偶联蛋白2(UCP2)、过氧化物酶增殖体激活受体α(PPAR-α)、过氧化物酶增殖体激活受体γ(PPAR-γ)、类胰岛素样生长因子 (IGF-Ⅰ)的mRNA丰度, 探讨在饲料中添加酵母培养物dv、姜黄素和水飞蓟素后, 草鱼肝胰脏脂代谢相关基因表达丰度的变化。结果显示, TAA诱导草鱼肝损伤后, FAS基因表达丰度上调, 调控脂质分解代谢的上游基因(如PPAR-α、ΡΡΑR-γ、PGC1-α)表达丰度没有显著性的变化, 而SCD1基因表达丰度显著性下调, 其结果可能导致脂质在肝细胞积累量增加, 诱发脂肪性肝病的发生和发展。在饲料中添加酵母培养物dv、姜黄素和水飞蓟素后, 在一定程度上修复了TAA对肝胰脏脂代谢的损伤。     

Dietary taurine can improve the hypoxia-tolerance but not the growth performance in juvenile grass carp Ctenopharyngodon idellus

This study was conducted to evaluate the effects of dietary taurine, as a feed additive, on the hypoxia-tolerance and growth performance of the juvenile grass carp Ctenopharyngodon idellus, one of the most important and intensively cultured freshwater fish, with the largest production in China. Graded levels of taurine (0, 0.5, 1, 1.5, 2 and 2.5 g kg^–1 dry diet) were fed to grass carp juveniles (mean weight: 5.26 ± 0.03 g) for 8 weeks. The survival time during acute hypoxia increased as dietary levels of taurine increased, with the highest dose of taurine resulting in the best acute hypoxia-tolerance. The erythrocyte osmotic fragility in grass carp was significantly improved when dietary taurine level was at least 1.5 g kg^?1 diet and can be improved much more when dietary taurine level was up to 2.5 g kg^?1 diet. A significant correlation between hemolysis rate of the erythrocyte osmotic fragility test and the survival time of acute hypoxia (r = ?0.873, P = 0.023 < 0.05) strongly suggested that the biomembrane stabilization function of taurine may contribute to its role of enhancing acute hypoxia-tolerance in grass carp. Dietary taurine cannot improve growth performance of grass carp, but it can increase the value of mesenteric fat index, indicating that dietary taurine influences the lipid metabolism. This study provides valuable information to improve hypoxia-tolerance of grass carp.     

Effects of low-voltage constant direct current on plasma biochemical profiles and gene expression levels in crucian carp <Emphasis Type="Italic">Carassius carassius</Emphasis>

This study investigated the plasma biochemical profiles and gene expression levels of crucian carp Carassius carassius subjected to anesthetization with low-voltage constant direct current (DC) and tricaine methanesulfonate (MS222). Fish in the MS222 treatments and the DC treatments were exposed to either two concentrations of MS222 (100 and 200 mg/l) or constant DC with a voltage of 26 V, at a voltage gradient of 0.68 V/cm, respectively. The results showed that low-voltage constant DC immobilized fish more quickly, resulting in significant elevations of plasma cortisol levels compared to exposure of high or low doses of MS222. Electronarcosis led to an increased expression of heat shock proteins 70 (HSP70) as well as HSP90 compared to the treatment group with MS222. Also, electronarcosis enhanced mitochondrial respiratory chain gene (atp6, cox1 and nd5) and antioxidant enzyme gene (CAT, SOD and GSH-px1) expression levels. In conclusion, low-voltage constant DC can disturb fish plasma biochemical profiles and gene expression levels.     

Growth,hematological and biochemical indices of common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> fed diets containing corn gluten meal

The effects of replacing fish meal (FM) with corn gluten meal (CGM) on growth and physiological performance were evaluated in common carp (Cyprinus carpio). Six experimental diets were formulated by substituting 0 (control), 20, 40, 60, 80, and 100% CGM protein for FM protein. The concentrations of dietary protein in the experimental diets were 27.8–29% and the P:E ratios were 14.7–15.46 mg/kJ. Eighteen fish with an initial weight of 13.5?±?0.1 g were allocated to each of 18 circular tanks (450 L) to give triplicate groups for each dietary treatment. The fish were fed to satiation for 8 weeks. At the end of the feeding trial, growth indices, body proximate composition, and hematological and biochemical parameters were measured. Blood samples were taken from six fish in each tank. Final weight and total length were significantly higher in fish fed 100% CGM (27.8?±?1.2 g and 11.9?±?0.3 cm) than for those fed the control (22.7?±?1.4 g and 10.9?±?0.5 cm) or 20% CGM (22.3?±?1.2 g and 11?±?0.4 cm) diets. No effect of FM replacement by CGM was observed for condition factor or hepatosomatic index (P?>?0.05). The highest value of protein productive value (14.31?±?0.65) was observed in fish fed 20% CGM (P?<?0.05). There were no significant differences in percentage body moisture and fat, but percentages of protein and ash were significantly different among experimental groups; the highest values of protein (15.6?±?0.24%) and ash (3.01?±?0.26%) were recorded in fish fed 40% CGM. For hematological parameters, the highest number of white blood cells (4.1?±?0.1?×?10³ mm^?3) was observed in fish fed 100% CGM (P?<?0.05). In addition, the highest hematocrit (42.1?±?0.7%) and triglyceride (294.11?±?23.82 mg dl^?1) were seen in fish fed the diet containing 40% CGM, while 80% CGM gave the highest cholesterol level (204.44?±?9.0 mg dl^?1; P?<?0.05). Replacement of FM with CGM had no negative effects on growth and physiological parameters of common carp fingerlings in this short (8 weeks) trial, suggesting that it may be feasible to replace FM with CGM in diets formulated for juvenile common carp.