Nutritional and biochemical studies on feeding of hydrolysed and unhydrolysed detoxified Jatropha curcas protein isolate in common carp fingerlings

Detoxified Jatropha protein isolate (DJPI), prepared from Jatropha seed cake was hydrolysed using Alcalase 2.4L FG. Both DJPI and hydrolysed DJPI (HDJPI) were evaluated as a fish meal (FM) replacement in common carp (Cyprinus carpio L.) diets. The peptide concentration in HDJPI was determined using matrix‐assisted laser desorption ionization‐time of flight mass spectrometer (MALDI‐TOF). Twenty‐two peptides with molecular mass ranging from m/z 906.455 to 1774.943 Da were identified. Fingerlings (45; av. wt. 10.1 ± 0.4 g) were randomly distributed in five groups with three replicates. An 8‐week feeding experiment was conducted to compare the nutritional quality of the HDJPI, DJPI and FM. Fish were fed on iso‐nitrogenous diets: control (FM based protein), J₅₀ (50% of FM protein replaced by DJPI), J₇₅ (75% of FM protein replaced by DJPI), JH₅₀ (50% of FM protein replaced by HDJPI) and JH₇₅ (75% of FM protein replaced by HDJPI). Growth performance was higher for FM fed group compared with the other groups, while nutrient utilization parameters were similar for control and J₅₀ groups and superior (P < 0.05) to J₇₅ and both the HDJPI fed groups. No variation (P > 0.05) was observed in blood albumin and hepatosomatic index among all different groups. Blood urea nitrogen, calcium, sodium and potassium ions in the blood were similar among all five groups.     

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.     

Haemato-biochemical,non-specific immunity,antioxidant capacity and histopathological changes in <Emphasis Type="Italic">Labeo rohita</Emphasis> fingerlings fed rubber protein isolate

A 60-day feeding trial was conducted to evaluate the haemato-biochemical, innate immune response, antioxidant capacity and histopathological changes in Labeo rohita fingerlings fed rubber protein isolates (RPI). One hundred and eighty fingerlings (average weight 4.45 ± 0.01 g) were distributed into five experimental groups in triplicate and fed with isonitrogenous and isocaloric diets. Soybean protein isolate (SPI) served as the reference diet (Control), and the treatment diets were formulated as RPI₂₅, RPI₅₀, RPI₇₅ and RPI₁₀₀ replacing 25, 50, 75 and 100% of SPI protein, respectively. The growth performance indices like final body weight (9.54–10.27 g), net weight gain (5.09–5.84 g), metabolic growth rate (4.54–5.02) and feed efficiency ratio (0.60–0.65) among the various groups were not significantly different (P > 0.05). All the haematological parameters, except red blood cells, showed no significant differences compared with the control group (P > 0.05). The immuno-biochemical parameters like albumin, globulin, total immunoglobulin, respiratory burst and lysozyme activities among the various groups did not differ significantly (P > 0.05). The stress enzyme such as superoxide dismutase (SOD), catalase (CAT) and oxidative stress marker malondialdehyde (MDA) showed no significant difference (P > 0.05). Histopathological examination of the liver revealed no marked changes. In summary, the results showed that RPI was well utilised by the fish and its inclusion did not generate any oxidative-induced stress, thus, RPI may be suggested as a potential replacement for SPI in fish diets without any detrimental effects. Hence, protein isolation offers a unique opportunity for the utilisation of rubber seed meal.     

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.     

Assessing the efficacy of three methionine sources in low protein and low fish meal diet for Chinese soft-shelled turtle, <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>

An 8-week feeding trial was conducted to evaluate the effects of three dietary methionine (Met) sources [dl-Met, coated-Met, and a methionine hydroxy analogue calcium salt (MHA-Ca)] for Chinese soft-shelled turtle (Pelodiscus sinensis). Triplicate groups of juvenile turtles (initial weight 3.48 ± 0.03 g) were fed twice per day at 3% of body weight with positive control diet (T1, 46% protein and 46% fish meal, FM), negative control diet (T2, 43% protein and 23% FM), or three other test diets supplemented with either 0.2% coated-Met (T3), 0.125% MHA-Ca (T4), or 0.1% dl-Met (T5) to the T2 basal formulation, respectively. The feeding trial was conducted in 15 350-L plastic containers with three replicates per dietary treatment. The results showed that the highest and lowest percentage weight gain (1023.5 ± 18.2 versus 882.1 ± 14.5%) and feed efficiency (87.2 ± 0.94 versus 81.4 ± 0.4%) were observed in turtles fed the T1 and T2 diet (P < 0.05), respectively. Significant improvement in weight gain was observed in turtles fed diets with Met supplementation, irrespective of source, compared with turtles fed the T2 diet. It was observed that dietary MHA-Ca had similar efficacy compared with dl-Met in terms of growth performance and feed utilization efficiency of the turtles (P > 0.05). Protein utilization efficiency was significantly higher in turtles fed T4 or T5 diet compared to the T2 diet. Whole body protein content (17.0 to 17.4% on a wet weight basis) in turtles fed with Met-added diets was comparable to those of turtles fed the T1 diet which were significantly higher compared to T2-fed turtles. Antioxidant defense system enzymes, superoxide dismutase, and glutathione peroxidase showed the highest activity, 658.8 ± 17.9 U/mL and 642.8 ± 17.5 μmol/L, respectively, in the serum of turtle fed the MHA-Ca supplemented diet and was significantly higher compared to turtles fed the T2 or T3 diet. The present results showed that P. sinensis are able to effectively use added MHA-Ca and dl-Met in low protein and low FM diets to enhance growth, feed utilization efficiency, nitrogen retention, and antioxidant defense system enzyme activities.     

Galactooligosaccharide and a combination of yeast and β-glucan supplements enhance growth and improve intestinal condition in striped catfish <Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis> fed soybean meal diets

The effects of galactooligosaccharide (GOS) and a combination of yeast and β-glucan (YβG) supplementation of dietary soybean meal (SBM) on the growth and digestive performance of striped catfish Pangasianodon hypophthalmus were evaluated. Four isonitrogenous (30% protein) and isocaloric (19 MJ/kg diet) diets were formulated to contain 100% fish meal (FM) protein, 55% FM protein/45% SBM protein, FM-SBM supplemented with 1% GOS, and a combination of 1% yeast and 0.1% β-glucan, respectively. Each diet was fed for 12 weeks to three groups of 30 striped catfish fingerlings (average weight 16.45?±?0.07 g) maintained in circular fiberglass tanks (600 l). Growth, feed utilization, and muscle protein composition of fish improved significantly after supplementation with either GOS or YβG compared to the unsupplemented SBM diet, but were similar to those of fish fed the FM diet. Nutrient digestibility, digestive enzyme activities, villi and microvilli length were significantly increased in fish fed the supplemented SBM diets. The gut microbiota ranking profile showed that supplementing the SBM diet with YβG and GOS gave a ranking of Verrucomicrobia, Spirochaetes, Bacteriodetes, and Actinobacteria phyla similar to that of the FM diet. Thus, diet containing 45% protein from soybean supplemented with either GOS or YβG can be recommended to improve the growth and digestive performance of striped catfish.     

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.     

Growth performance and biochemical composition of juvenile pearl oyster <Emphasis Type="Italic">Pinctada martensii</Emphasis> fed on artificial diets

The effects of three artificial diets (S1, S2 and S3) on survival, growth and biochemical composition of one-year-old pearl oyster Pinctada martensii were investigated. Six experimental groups (EG1, EG2, EG3, EG4, EG5 and EG6) and one control group (CG) were set up. EG1, EG2 and EG3 were solely fed on S1, S2 and S3, respectively. EG4, EG5 and EG6 were fed on mixed diets, as follows: S1 and Platymonas subcordiformis; S2 and P. subcordiformis; and S3 and P. subcordiformis, respectively. CG was fed on only P. subcordiformis. All groups were continuously fed for 60 days. Survival, growth and biochemical composition of soft tissues were compared across the groups. Results showed that survival rate, the absolute growth rate (AGR) and relative growth rate (RGR) of shell length did not differ significantly across the groups (p > 0.05). The AGR and RGR of total weight differed significantly among the groups (p < 0.05). AGR and RGR of shell length and total weight were the highest in EG5 and the lowest in EG1. Gross fat content showed insignificant differences among the groups (p > 0.05). However, gross protein content and ash content showed significant differences across the groups (p < 0.05). The gross protein of the groups solely fed on artificial diets was lower than those of the groups fed on mixtures of artificial diets and microalgae or single microalgae. The contents of other amino acids, total amino acids (TAAs) and essential amino acids (EAAs) showed significant differences across the groups (p < 0.05). TAA, EAA and delicious amino acids of the groups fed solely on artificial diets were lower than those of the groups fed on mixtures of artificial diets and microalgae or single microalgae. Results indicated that the artificial diet (S2) can serve as substitutes of microalgal diets for P. martensii.     

Growth performance and immune responses of gibel carp, <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>, fed with graded level of rare earth-chitosan chelate

A 12-week feeding trail was conducted to assess the effect of rare earth-chitosan chelate (RECC) on growth performance and immune responses of gibel carp, Carassius auratus gibelio. Isonitrogenous and isolipid experimental diets were supplemented with graded levels of rare earth-chitosan chelate (RECC 0, 0.8, 4 and 8 g Kg^?1). A total of 720 gibel carps (initial body weight about 14.32 g) were divided randomly into four groups with six replicates, respectively. Fish were fed with the experimental diets three times every day. At the end of the feeding trail, the survival rate was higher than 96 %. Weight gain rate and specific growth rate (SGR) significantly increased with RECC supplementation up to 0.8 g Kg^?1 (P < 0.05) and tended to decline at higher supplementation levels, while feed conversion ratio was not significantly different between groups (P > 0.05). Based on broken-line regression analysis of SGR, the optimum dietary RECC was estimated to be 0.71 g Kg^?1 of the diet. Condition factor and viscerosomatic index were not significantly affected by RECC (P > 0.05), while hepatosomatic index in the group fed with 0.8 g Kg^?1 RECC (3.45 ± 0.10 %) was significantly lower than in other groups (P < 0.05). Plasma ALT was significantly affected by RECC (P < 0.05), while AST was not. Plasma total protein and albumin were increased with RECC supplementation up to 0.8 g Kg^?1 and decreased significantly at higher supplementation levels (P < 0.05). RECC supplementation significantly decreased plasma urea and glucose concentration and increased plasma creatinine concentration significantly (P < 0.05). Respiratory burst activity of phagocytes and myeloperoxidase activity were not significantly different between groups, while superoxide dismutase activity and nitrogen monoxide concentration were increased with the increasing level of RECC in the diets. In conclusion, RECC could enhance growth performance and improve immunity of gibel carp.     

Effects of dietary yeast inclusion and acute stress on post-prandial whole blood profiles of dorsal aorta-cannulated rainbow trout

Yeast is a potential alternative to fish meal in diets for farmed fish, yet replacing more than 50 % of fish meal results in reduced fish growth. In a 4-week experiment, 15 rainbow trout (Oncorhynchus mykiss) were cannulated and fed three diets each week: 30 % fish meal as a control (FM); 60 % replacement of fish meal protein, on a digestible basis, with Saccharomyces cerevisiae (SC); and 60 % replacement with Wickerhamomyces anomalus and S. cerevisiae mix (WA). Blood was collected at 0, 3, 6, 12 and 24 h after feeding. In the final week, fish were exposed to a 1-min netting stressor to evaluate possible diet–stress interactions. Significant increases in pH, TCO₂, HCO₃ and base excess were found after fish were fed the SC and WA diets compared with FM, which elevated blood alkaline tides. Yeast ingredients had lower buffering capacity and ash content than fish meal, which explained the increase in alkaline tides. In addition, fish fed the WA diet had significantly reduced erythrocyte area and fish fed SC and WA diets had increased mean corpuscular haemoglobin levels, indicating haemolytic anaemia. Higher levels of nucleic acid in yeast-based diets and potentially higher production of reactive oxygen species were suspected of damaging haemoglobin, which require replacement by smaller immature erythrocytes. Acute stress caused the expected rise in cortisol and glucose levels, but no interaction with diet was found. These results show that replacing 60 % of fish meal protein with yeasts can induce haemolytic anaemia in rainbow trout, which may limit yeast inclusion in diets for farmed fish.     

Utilization of a by-product from the Jatropha biodiesel industry as a fish meal replacer in common carp Cyprinus carpio L. diets

Detoxified Jatropha kernel meal (DJKM) was created through heat and solvent extraction and replaced fish meal (FM) at 0% (C_ontrol), 50% (J₅₀), or 62.5% (J_62.5) in common carp Cyprinus carpio fingerling (initial mean weight ± SD; 22 ± 0.12 g) diets over 16 weeks. Growth performance and feeding efficiencies were similar (P > 0.05) among groups, but nutrient digestibilities and digestive enzyme activities were significantly higher (P < 0.05) in C_ontrol than others. The slightly lower growth and significantly lower nutrient digestibility in carp fed DJKM indicates a need to improve the nutritive value of DJKM as a promising and sustainable FM alternative.     

Growth,enzymatic glutathione peroxidase activity and biochemical status of juvenile barramundi (<Emphasis Type="Italic">Lates calcarifer</Emphasis>) fed dietary fermented soybean meal and organic selenium

Solvent-extracted soybean meal (SBM) was fermented using baker’s yeast Saccharomyces cerevisae at 30 °C for 5 days. Four isonitrogenous and isocaloric diets containing 75% SBM protein, either fermented or non-fermented (SBM and FSBM), and supplemented or not with organic Se (OS) (SBM_OS and FSBM_OS), were fed to triplicate groups of juvenile barramundi (Lates calcarifer) (initial weight of 5 g) for 75 days. A fishmeal (FM)-based diet formulated for juvenile barramundi was used as a reference diet. The growth of fish was significantly affected by either the interaction of SBM type or by the OS level. In fish fed diets supplemented with OS (SBM_OS and FSBM_OS), final weight (FW), specific growth rate (SGR) and weight gain (WG) were higher in fish fed the fermented SBM (FSBM_OS) than in those fed the non-fermented SBM (SBM_OS). The apparent digestibility coefficient (ADC) of protein was higher in the fish fed the fermented SBM, either supplemented or unsupplemented with OS. However, there were no significant differences in the ADC of dry matter (DM) and lipids among the tested diets and in comparison to the reference diet. The haematocrit and leucocrit of fish fed the FSBM_OS diet were lower than those of fish fed the FM diet. Furthermore, glutathione peroxidase (GPx) activity was significantly influenced by OS supplementation in the experimental diets; GPx activity was greater in the fish fed diets supplemented with OS. Creatinine kinase (CK) of all groups of fish was higher than the CK of those fed the reference diet. These results suggest that with a proper nutritional level, OS supplementation may act as an important factor in enzymatic GPx activity and in the haematology and blood biochemistry status of juvenile barramundi fed fermented SBM-based diets, encouraging improvement of the overall growth performance.     

Hepatic RNA:DNA ratio as a reliable indicator of the growth performance of juvenile <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis> under nutritional stress

We investigated the reliability of the ratio of RNA to DNA in juvenile Pelodiscus sinensis under moderate nutritional stress from lack of sufficient protein. The control diet contained 45% protein. Two experimental diets containing 40 and 35% protein were used to trigger nutritional stress. One hundred fifty-six turtles were randomly divided into the three diet groups and fed for 90 days. Four turtles were randomly sampled from each group on days 2, 4, 7, 15, 30, 45, 60, 75 and 90 of the trial and their hepatic RNA:DNA was measured. Each turtle was weighed on days 15, 30, 45, 60, 75 and 90. Obvious positive correlations between RNA:DNA and specific growth rate (SGR) were found, which were affected by dietary protein levels. The exponential regression model of the relationship between RNA:DNA (x) and SGR (y) in turtles fed 45% protein diet was y = 0.8001e^0.3072x , in the 40% protein diet it was y = 0.5374e^0.4297x , and in the 35% protein diet it was y = 0.7867e^0.369x . From day 7 until the end of the trial, RNA:DNA accurately distinguished differences among the three groups. We suggest that hepatic RNA:DNA is a sensitive and quick indicator of growth performance in juvenile turtles, especially in studies related to nutritional stress.     

Stimulatory effect of dietary taurine on growth performance,digestive enzymes activity,antioxidant capacity,and tolerance of common carp, <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L., fry to salinity stress

The present study was carried out to evaluate the effect of dietary taurine (Tau) on performance, digestive enzymes, antioxidant activity, and resistance of common carp, Cyprinus carpio L., fry to salinity stress. Fish (0.97?±?0.033 g) were fed on different taurine levels of 0.0 (control), 5, 10, 15, or 20 g/kg diet up to satiation twice daily for 8 weeks. At the end of the feeding trial, fish were stressed by exposure to 10 ppt salinity for 3 days during which fish mortality was observed. Fish performance was significantly (P?<?0.05) improved by dietary taurine up to 15 g Tau/kg diet after which fish growth and feed intake were almost the same. Also, taurine supplementation significantly (P?<?0.05) elevated activities of intestinal amylase, lipase, and protease resulting in an improving in feed intake giving better performance. Furthermore, Tau-stimulated antioxidant activity of common carp was observed in a dose-related manner, where activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly (P?<?0.05) higher, but malondialdehyde (MDA) value was significantly (P?<?0.05) lower in Tau-fed fish groups than those fed the control diet. In salinity stress experiment, highest survival rate was observed at fish fed Tau-supplemented diets without significant (P?>?0.05) differences over fish fed the control diet. It appears that taurine could be used as a feed supplement to confer better growth and health of common carp fry with optimal level of 15 g/kg diet.     

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.     

Silymarin inhibits adipogenesis in the adipocytes in grass carp <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> in vitro and in vivo

In this study, two experiments were performed to explore the function of silymarin in adipogenesis in grass carp (Ctenopharyngodon idellus) using in vitro and in vivo models. In experiment 1, differentiated grass carp pre-adipocytes were treated with silymarin for 6 days. Treatment with 100 μg mL^?1silymarin (SM100 group) significantly reduced triglyceride accumulation at day 6. The adipogenic gene expression levels of PPARγ, C/EBPα, SREBP1c, FAS, SCD1, and LPL, and the protein expression level of PPARγ were significantly down-regulated in the SM100 group. Additionally, the SM100 group had significantly lower reactive oxygen species production and reduced glutathione contents compared with the control in vitro. In experiment 2, the juvenile grass carp (mean body weight= 27.4 ± 0.17 g) were fed six isonitrogenous and isocaloric diets in a factorial design containing 0, 100, or 200 mg kg^?1 silymarin (SM0, SM100, SM200) associated with either 4 or 8% lipid levels (low lipid, LL, and high lipid, HL, respectively) for 82 days. The results demonstrated that dietary silymarin supplementation significantly reduced the elevated intraperitoneal fat index in grass carp fed with high-lipid diets, and the gene expression of adipogenesis (PPARγ, FAS) when supplemented with dietary silymarin was notably lower than when no silymarin was supplemented under the high-lipid diets. Thus, our data suggest that silymarin suppressed lipid accumulation in grass carp both in vitro and in vivo, and the effect might be due to an influence on the expression of adipogenesis factors and ROS production partly associated with effects on antioxidant capability.     

Evaluation of different <Emphasis Type="Italic">Pyropia yezoensis</Emphasis> extracts as feed additives for growth and immunity of Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

We evaluated the effects of diets incorporating the red algae Pyropia yezoensis, prepared by several different extraction methods, on the growth of juvenile Japanese flounder Paralichthys olivaceus. We assessed growth performance, as well as the levels of amino acids, fatty acids, insulin-like growth factor I (IGF-I) and interleukins (ILs). Four experimental diets were developed based on different methods of processing P. yezoensis. A commercial feed, laver powder (P), high-pressure heat extraction of laver (HPHE) and acid hydrolysis extraction of laver were used as the experimental diets. Three experimental replicates were established for each diet (40 fish/group, body weight 123.7 ± 1.1 g), and the fish were fed for 6 weeks. We found no significant differences in weight gain, specific growth rate or feeding efficiency among the groups (P > 0.05); however, the fish fed HPHE had the greatest growth performance. Fish fed the laver extracts exhibited the highest protein efficiency ratio compared with the control and P groups. The experimental groups fed P. yezoensis extracts had significantly higher levels of IGF-I (P < 0.05) than those of the control group. High levels of IL-2 were found in the P and HPHE groups, IL-12 in the HPHE group, and IL-6 in all experimental groups. Therefore, these results suggest that a P. yezoensis extract improved the growth performance and immunity of Japanese flounder. In particular, the high-pressure heating process was a useful extraction method for preparing a P. yezoensis extract, which had beneficial effects as a dietary supplement in Japanese flounder.     

Effects of starvation and feeding on blood chemistry,fatty acid composition and expression of vitellogenin and fatty acid-binding protein genes in female swimming crab <Emphasis Type="Italic">Portunus trituberculatus</Emphasis> broodstock

Portunus trituberculatus broodstock were stocked in plastic tanks to evaluate the effects of starvation and feeding on gonadal development, blood chemistry, fatty acid composition, and expression of vitellogenin (Vtg) and fatty acid-binding protein genes (FABP) in females. Two treatments (starved and fed) were randomly assigned to triplicate groups of 90 swimming crab broodstock (approximately 230 ± 45 g). In the starved treatment, crabs were starved for 30 days, whereas in the fed treatment crabs were fed once a day with clams. The gonadosomatic index decreased significantly in starved crabs (P < 0.05), as did the serum glucose and cholesterol concentrations; conversely, the total protein concentration in serum significantly increased (P < 0.05). In the ovary, there was a significant relative decline of 18:0, 16:1n-7 and 20:1n-9 fatty acids and relative increases of 20:4n-6, 22:6n-3, 18:1n-9 and 20:5n-3 in starved crabs compared to fed crabs (P < 0.05). Relative expression of Vtg in the ovary decreased significantly in starved crabs (P < 0.05), while there was no significant difference in hepatopancreas Vtg expression between starved and fed crabs (P > 0.05). Starvation suppressed gonadal development in female swimming crab broodstock.     

The effects of starvation and re-feeding on growth and swimming performance of juvenile black carp (<Emphasis Type="Italic">Mylopharyngodon piceus</Emphasis>)

We investigated the effects of starvation and re-feeding on growth and swimming performance and their relationship in juvenile black carp (Mylopharyngodon piceus). We measured the specific growth rate (SGR), resting metabolic rate (RMR) and constant acceleration test speed (U _CAT, the maximum swimming speed at exhaustion by constant acceleration test with 0.1667 cm s^?2 rate) in a treatment group (21 days of starvation then 21 days of re-feeding) and control group (routine feeding) (n = 20). Starvation resulted in a 17 % decrease in body mass of black carp (P < 0.05). After 21 days of re-feeding, body mass was greater than that of pre-starvation but still less than that of the control group at 42 days. During the re-feeding phase, the SGR of the treatment group was higher than that of the control group (P < 0.05). Starvation resulted in a significant decrease in the RMR and U _CAT. After 21 days of re-feeding, both the RMR and U _CAT recovered to the pre-starvation levels. In the control group, individual juvenile black carp displayed strong repeatability of the RMR and U _CAT across the measurement periods (P ≤ 0.002). In the treatment group, RMR showed significant repeatability between pre-starvation and re-feeding (P = 0.007), but not between pre-starvation and starvation or between starvation and re-feeding. U _CAT showed significant repeatability between pre-starvation and starvation (P = 0.006) and between pre-starvation and re-feeding (P = 0.001), but not between starvation and re-feeding. No correlation or only a weak correlation was found between any two variables of RMR, U _CAT and SGR, whereas the increment of the U _CAT (ΔU _CAT) was negatively correlated with that of SGR during the starvation phase (r = ?0.581, n = 20, P = 0.007) and re-feeding phase (r = ?0.568, n = 20, P = 0.009). This suggested that within individual black carp, there is a trade-off between growth and maintenance (or development) of swimming performance under food-limited conditions.     

Dietary inclusion of detoxified <Emphasis Type="Italic">Jatropha curcas</Emphasis> kernel meal: effects on growth performance and metabolic efficiency in common carp, <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.

Jatropha curcas is a multipurpose and drought-resistant shrub or small tree widespread all over the tropics and subtropics. Its seeds are rich in oil, and the Jatropha kernel meal obtained after oil extraction is rich in protein. However, presence of toxic and antinutritional constituents restricts its use in fish feed. Jatropha kernel meal was detoxified. Common carp, Cyprinus carpio, fingerlings (15; av. body mass 10.9 ± 0.65 g) were randomly distributed in three groups with five replicates. A 6-week feeding experiment was conducted in a respirometer system to evaluate the growth performance, nutrient utilisation and energy budget. Fish were fed isonitrogenous diets (38% crude protein): control diet (C _ontrol group) containing fish meal (FM) protein based protein and two other diets replacing 75% FM protein with detoxified Jatropha kernel meal (DJKM, J _atropha group) and soybean meal (SBM, S _oybean group). At the end of the experiment, body mass gain, metabolic growth rate, protein efficiency ratio, protein productive value, energy retention, efficiency of metabolised energy for growth and efficiency of energy retention were determined. These parameters were high and statistically similar for C _ontrol and J _atropha groups and significantly lower (P < 0.05) for S _oybean group. Whereas a reverse trend was observed for energy expenditure per g protein retained in fish body. No significant differences were found in heat released, gross energy uptake, metabolised energy intake, metabolisability, energy expenditure, energy expenditure per g protein fed and apparently unmetabolised energy. Conclusively, common carp–fed plant protein (DJKM and SBM) and FM protein–based diets exhibited equal average metabolic rate.