Growth and feed performance,digestibility and acute stress response of juvenile grouper (Epinephelus fuscoguttatus) fed diets with hydrolysate from milkfish offal

Nutritional qualities of fish processing by‐products can further be improved through enzymatic hydrolysis. The objective of this study was to elucidate the efficacy of hydrolysed milkfish offal at different inclusion levels when fed to juvenile grouper, Epinephelus fuscoguttatus, with an initial body weight of 2.88 ± 0.06 g. The animals were fed for 56 days with seven diets supplemented with 0 (control), 5%, 15% and 25% of milkfish offal (MO) and milkfish offal hydrolysate (MOH). The diets were formulated to be isonitrogenous (45%) and isolipidic (11%). The diets were assigned to 21 tanks (15 fish per tank) with each diet having three replicates. Results from the experimental trials indicated that feed conversion efficiency, feed intake and weight gain of fish significantly (P < 0.05) improved when fed diets with MOH. No significant differences within the rest of the dietary treatments were observed. Survival rate (>90%) did not differ in all the dietary treatments. Proximate composition (crude protein, crude fat and ash) indicated no significant difference among fish fed from all the dietary treatments. Apparent digestibility of MOH indicated a 95% and 66% digestibility for protein and dry matter respectively. Plasma stress parameters (cortisol and glucose) were not influenced by the dietary treatment when fish were subjected to an acute stressor (5‐min chasing). Liver morphology indicated normal hepatocyte shape and the presence of lipid droplets in fish fed from all the dietary treatments. The results indicated that milkfish offal processed as hydrolysate can be utilized in grouper diets and can promote growth and feed efficiency when supplied at 10–15%.     

Effects of dietary coated protease on growth performance,feed utilization,nutrient apparent digestibility,intestinal and hepatopancreas structure in juvenile Gibel carp (Carassius auratus gibelio)

This study was conducted to investigate the effects of dietary protease on growth performance, feed utilization, whole‐body proximate composition, nutrient digestibility, intestinal and hepatopancreas structure of juvenile Gibel carp, Carassius auratus gibelio (mean weight 8.08 ± 0.18 g). Six diets were prepared, including a positive control diet (dietary protein 350 g/kg, PC), one negative control diet (dietary protein 33 g/kg, NC) and four protease supplementations diets, which were 75, 150, 300 and 600 mg/kg protease NC diet. After 12 weeks of diet feeding in indoor recycle aquarium tanks, no significant difference (p > .05) was found on growth performance between fish fed diet with 75–600 mg/kg protease and the PC group. Compared with the fish fed the NC diet, the specific growth rate of fish fed 300 mg/kg protease increased significantly (p < .05), as well as protein efficiency ratios (p < .05), while feed conversion was the opposite (p < .05). The nutrient digestibility of crude protein and lipid was higher (p < .05) in fish fed 150 mg/kg protease diet than the PC diet. Whole‐body proximate composition of fish was not affected (p > .05) by the dietary treatment. Serum alkaline phosphatase and albumin were significantly affected by dietary protease (p < .05), while the content of total protein, glucose, triglyceride, total cholesterol, aspartate aminotransferase and alanine aminotransferase activities in serum was not affected (p > .05). Foregut muscular thickness was thinner (p < .05), when the fish fed diets supplementation of protease in 150 or 600 mg/kg diet than the NC diet. Protease activities in hepatopancreas and foregut were higher (p < .05), in the fish fed 150 or 300 mg/kg protease diet than the fish fed the PC diet, but those in the mid‐ and hindgut were not significantly affected (p > .05) by the dietary treatments. Based on the regression analysis of weight gain rate, the optimal dietary inclusion level of protease was 400 mg/kg in the diet for juvenile Carassius auratus gibelio.     

The optimal dietary protein level of juvenile silver sillago Sillago sihama at three dietary lipid levels

A 5 × 3 factorial growth trial was conducted to evaluate optimal dietary protein and lipid levels (dietary protein level, DP; dietary lipid level, DL) for juvenile Sillago sihama (S. sihama) (2.0 ± 0.02 g, initial weight). Fish were fed 15 diets containing 5 DPs (350, 400, 450, 500 and 550 g/kg) and 3 DLs (60, 90 and 120 g/kg) for 8 weeks. The interaction between proteins and lipids significantly influenced the feed conversion ratio, condition factor, body composition, antioxidant indices and lipase activity (p < .05). DP 450 g/kg showed the highest average final body weight. DPs 500 and 550 g/kg significantly decreased the protein efficiency ratio (p < .05). DL 120 g/kg showed the highest percentage weight gain. The low feed conversion ratio was found in diets P45L12, P55L9 and P55L12. Diet P45L12 showed high superoxide dismutase activities. DP 450 g/kg showed the lowest average malondialdehyde content. Lipase activity was increased by increasing DP (p < .05) with a fall at DP 550 g/kg. Under the present experimental conditions, the optimal DP for S. sihama was 450 g/kg under the DL 120 g/kg.     

Influence of dietary protein level on growth performance,digestibility and activity of immunity‐related enzymes of leopard coral grouper,Plectropomus leopardus (Lacépède, 1802)

An 8‐week feeding trial was conducted to assess the interaction between dietary protein levels and fish growth, digestibility and activity of immunity‐related enzymes of Plectropomus leopardus. Five diets with different protein levels (400 g/kg, 450 g/kg, 500 g/kg, 550 g/kg and 600 g/kg protein) were designed. P. leopardus fed with 500 g/kg, 550 g/kg and 600 g/kg dietary protein, showed higher weight gain rates than fish fed 400 g/kg and 450 g/kg dietary protein. Ingestion rate in fish fed with 500 g/kg dietary protein was significantly higher than those with other diets. P. leopardus fed with 500 g/kg, 550 g/kg and 600 g/kg dietary protein, showed that feed coefficients were significantly lower than those fed with 400 g/kg and 450 g/kg dietary protein. Net protein utilization was significantly lower in fish fed with 400 g/kg diet than those with other diets. Fish fed with 400 g/kg and 450 g/kg dietary protein had an apparent feed digestibility coefficient for dry matter that was significantly lower than that with other diets. Protease activity was highest in fish fed on 500 g/kg dietary protein. Fish fed with 500 g/kg dietary protein, had the highest superoxide dismutase activity. Fish fed with 600 g/kg dietary protein, had the highest alkaline phosphatase activity. Thus, a diet containing 500 g/kg protein is recommended for P. leopardus aquaculture.     

Partial substitution of soybean meal with fermented soybean residue in diets for juvenile largemouth bass,Micropterus salmoides

A feeding trial was conducted to investigate the effects of partial replacement of soybean meal (SBM) with fermented soybean residue (FSR) on growth performance, body composition and plasma biochemical parameters of largemouth bass, Micropterus salmoides. Soybean residue was fermented with a mixture of microorganisms (Bacillus subtilis, Lactobacillus spp. and Molasses yeast) using the solid‐state fermentation. Four isonitrogenous (crude protein 430 g/kg) and isoenergetic (gross energy 18 MJ/kg) diets were formulated by replacing 0 (the control), 20, 40 and 60g/kg of protein from SBM with FSR (FSR0, FSR20, FSR40 and FSR60, respectively). Each diet was fed to four replicate groups of fish (initial body weight: 17.1 ± 0.19 g) for 12 weeks. Results showed that dietary FSR substitution significantly improved growth of juvenile largemouth bass. The weight gain, specific growth rate and protein efficiency ratio were all significantly improved by dietary FSR level up to 40g/kg substitution level (p < .05) and then levelled off beyond this level. Fish fed the diet with 40g/kg and 60g/kg protein from FSR had lower feed conversion ratio than the control group (p < .05). The hepatosomatic index, viscera ratio and liver lipid content significantly decreased with increasing dietary FSR level. Total protein content, superoxide dismutase and alkaline phosphates activities in plasma were lower in fish fed the control diet (p < .05) than the other groups. However, both alanine aminotransferase and aspartate transaminase were higher in fish fed the control diet (p < .05) compared to the other treatments. The plasma catalase activity significantly increased with increasing dietary FSR level, while plasma triglyceride, total cholesterol, glucose and malondialdehyde contents significantly reduced. No significant difference was observed in the glutathione peroxidase activity among dietary treatments. These findings demonstrated that replacing dietary SBM with FSR has beneficial effects on growth of M. salmoides, and the best growth performance was obtained at 40g/kg replacement for SBM protein. In addition, there is a great potential to apply FSR to improve lipid metabolism and antioxidant capacity of M. salmoides.     

Use of supplemental amino acids on the success of reducing dietary protein levels for Jian carp (Cyprinus carpio var. Jian)

A feeding trial was conducted to investigate the effects of essential amino acids (EAA) supplementation on the success of dietary protein reduction for Jian carp (Cyprinus carpio var. Jian). Five diets were formulated including a positive control (Diet 1, 378.4 g/kg protein), a negative control (Diet 5, 289.8 g/kg protein) and three test diets (Diet 2‐Diet 4, 340.4–299.0 g/kg protein supplemented with EAA). No significant decline of growth performance in fish fed decreased dietary protein with EAA supplementation, which were significantly higher than the negative control. The apparent digestibility coefficients (ADC) of protein and most EAAs were not significantly affected by the dietary treatments. Dietary treatments did not affect whole‐body composition, but influenced protein and EAAs retentions. Total ammonia nitrogen (TAN) excretion decreased with the decreasing dietary protein, and the highest plasma ammonia content was observed in fish fed Diet 5. Plasma glucose concentration increased with the decreasing dietary protein reduction, while other plasma parameters were not affected by dietary treatments. These results indicated that low‐protein diets (299.0 g/kg) supplemented with EAA can maintain protein, EAA digestibility and retention in Jian carp, and also has benefit in the reduction of TAN excretion.     

Dietary supplementation of red alga Pyropia spheroplasts on growth,feed utilization and body composition of sea cucumber,Apostichopus japonicus (Selenka)

A feeding experiment was conducted in a closed recirculating system to evaluate the effects of freeze‐dried spheroplasts prepared from Pyropia yezoensis (Ueda) on feed intake, growth and biochemical composition of sea cucumber, Apostichopus japonicus (Selenka). Pyropea spheroplasts (PS) were prepared through enzymatic treatment to break down the complex mixture of polysaccharides cell walls that might be easier for growth energy partitioning. Sea cucumbers were fed‐formulated diets with 10 (Diet 1), 30 (Diet 2) and 50 g/kg (Diet 3) inclusion level of PS. A diet without PS was used as a control (Diet 4). The experiment was conducted for 6 weeks maintaining water temperature 15 ± 1°C, photoperiod 18:06 hours (D:L). Feed was supplied ad‐libitum at 16.00 h once in a day, and the remaining feed and faeces were removed in the next day. Results showed that the highest growth was observed in the 50 g/kg PS diet compared to other treatments. Total weight gain, mean weight gain, net yield, protein efficiency ratio (PER) and protein gain (%) were significantly higher in the 50 g/kg PS diet (p < .05). A significantly higher percentage of energy was allocated for growth in the 50 g/kg PS diet. The highest specific growth rate and feed conversion efficiency (p < .05) were observed in the higher percentage of PS diet. Both the growth performance and biochemical analysis showed that superior growth was observed with increasing levels of PS in the diet. We infer that PS can be used as a new, cheaper feed ingredient in the formulated diet of A. japonicus.     

Effects of fish meal replacement with meat and bone meal using garlic (Allium sativum) powder on growth,feeding, digestive enzymes and apparent digestibility of nutrients and fatty acids in juvenile rainbow trout (Oncorhynchus mykiss Walbaum, 1792)

Effects of graded dietary levels of fish meal (FM) replacement with meat and bone meal (MBM) with garlic powder (G) or without garlic powder (WG) garlic powder were investigated on growth, digestive enzymes, apparent digestibility, body composition and fatty acid profile of juvenile rainbow trout (Oncorynchus mykiss; 8.26 ± 1.10 g). A total of eight dietary treatments were evaluated: control with garlic powder (0 MBM+G), 250 MBM+G, 450 MBM+G, and 650 MBM+G (g MBM/kg diet; treatments with garlic powder; G treatments), control without garlic powder (0 MBM), 250 MBM, 450 MBM and 650 MBM (g MBM/kg diet; treatments without garlic powder; WG treatments). Our results demonstrated that increased dietary content of MBM impaired growth and production performance, body composition, digestive enzyme activity, tissue fatty acid profile and overall digestibility. Addition of garlic powder was able to correct fish performance, body composition, enzyme activity and digestibility to some extent. Dietary supply of garlic resulted in increased digestive enzyme activity improving fish performance. The current results indicated that fish fed 650 g MBM/kg feed exhibited impaired growth that could be attributed to reduce digestibility of protein, fat, energy and dry matter, PUFAs and n‐3 LC‐PUFAs in muscle. We suggest that 450 g FM per kilo feed can be successfully replaced with MBM supplemented with 30 g garlic powder per kilo feed without impairing fish performance and physiology, however distorting tissue fatty acid composition resulting in lesser levels of n‐3 LC‐PUFAs compared to the 0 MBM+G diet.     

Fishmeal alternative from renewable CO2 for rainbow trout feed

The rapid global growth in fish farming and limited supply of fish meal (FM) has consequently reduced FM inclusion levels in compound feeds leading to a higher reliance on alternative protein sources. Sasya is a single cell protein (SCP) product that has a similar amino acid profile as FM. The aim of this study was to evaluate the effects of replacing FM with SCP on in vivo digestibility, growth, feed efficiency, whole‐body proximate/amino acid composition and gene expression levels of various hepatic enzymes in rainbow trout (Oncorhynchus mykiss). Three isonitrogenous (470 g/kg crude protein) and isolipidic (18 g/kg crude lipid) diets were formulated as follows: Diet 1: control (30% FM); Diet 2:24% FM + 6% SCP and Diet 3:18% FM + 12% SCP. Each diet was hand‐fed to triplicate tanks containing 30 rainbow trout fingerlings (4.99 ± 0.20 g) for 9 weeks. Apparent digestibility coefficients of SCP for dry matter, crude protein, lipid and energy were 60, 80, 93 and 74% respectively. Growth performance (final weight: 69–71 g), feed conversion ratios (0.91–0.94) as well as whole‐body protein and amino acid composition were unaffected by diets. However, Diet 3 significantly increased whole‐body crude fat and energy. Fish fed the SCP‐based diets had significantly higher expression for carnitine palmitoyltransferase‐1b (CPT1b), fatty acid delta 6 desaturase (FADS6) and fatty acid elongase 5 compared to the control. Overall, the quality of the SCP was similar as FM. Therefore, this product could enlarge the portfolio of alternative protein sources that can be used in fish diets and thus open a new market opportunity for use of a new feed resource in the feed industry.     

Effects of dietary carbohydrate to lipid ratio on growth,feed utilization,body composition and digestive enzyme activities of golden pompano (Trachinotus ovatus)

An 8‐week feeding trial was conducted to investigate the effects of dietary carbohydrate to lipid ratio (CHO: L) on growth, feed utilization, body composition and digestive enzyme activities of golden pompano, Trachinotus ovatus. Five iso‐nitrogenous (450 g/kg protein) and iso‐energetic (19 MJ/kg gross energy) diets with varying CHO: L ratios of 0.68, 1.02, 1.62, 2.61 and 4.35, respectively, were fed to triplicate groups of 30 fish (average 13.8 ± 0.1 g). Results showed that dietary CHO: L ratios did not show any significant influence on survival of golden pompano (p > .05) but significantly affected its growth performance and feed utilization (p < .05). Fish fed diets with CHO: L ratios at 1.62 and 2.61 exhibited the highest final body weight, weight gain ratio, specific growth rate, feed efficiency ratio and protein efficiency ratio. Fish body lipid and liver glycogen contents were also significantly influenced by CHO: L ratio (p < .05). Hepatic amylase activity increased firstly and then decreased as the dietary CHO: L ratio increased, while lipases activity decreased with increasing dietary CHO:L level. The regression model analysis showed that the most suitable dietary CHO: L ratio (protein 450 g/kg) to reach the highest weight gain ratio is 2.38.     

Growth performance,muscle and liver composition,blood traits,digestibility and gut bacteria of beluga (Huso huso) juvenile fed different levels of soybean meal and lactic acid

The objective of this study was to assess the effects of fishmeal (FM) replacement with 0, 350 or 700 g/kg soybean meal (SBM) in combination with the supplementation of lactic acid (LA; 0, 10 or 20 g/kg) in the diets of juvenile beluga sturgeon (Huso huso; 700 ± 30 g). Nine isonitrogenous (400 g/kg protein) and isoenergetic (18 MJ/kg) diets were fed to beluga ad libitum, three times a day, for 60 days. The results showed that replacing FM with SBM without LA significantly reduced fish growth; on the other hand, LA supplementation had positive effects on fish fed diets that FM was replaced by SBM (p < .05). Increasing SBM in the diet altered the fatty acid profiles of the fish, reducing long‐chain polyunsaturated fatty acids and the n‐3/n‐6 fatty acids. High amounts of SBM (700 g/kg) caused reductions in the haematocrit, glucose and cholesterol levels in the blood (p < .05). In addition, the digestibility of protein, fat, dry matter and phosphorus was reduced when replacing FM with SBM, however, adding LA to the diets increased fish performance (p < .05), and this improvement was sharper in 2% LA groups. The number of LA bacteria increased significantly with the dietary supplementation of LA (p < .05). Based on these results, replacing 350 g/kg of FM with SBM and adding 20 g/kg LA to their feed do not negatively affect the biological and physiological indices of beluga.     

Effect of replacing fish meal with meat and bone meal on growth,feed utilization and nitrogen and phosphorus excretion for juvenile Pseudobagrus ussuriensis

The effect of replacing fish meal (FM) with meat and bone meal (MBM) in diets for juvenile Pseudobagrus ussuriensis was evaluated in a 90‐day feeding trial. Six isonitrogenous (crude protein, 430 g/kg) and isolipidic (crude lipid, 74 g/kg) diets were formulated to contain MBM to replace FM at 0 (S0), 200 (S20), 400 (S40), 600 (S60), 800 (S80) and 1000 g/kg (S100), respectively. The results showed that there was no significant difference in weight gain (WG) among fish fed S0, S20 and S40 diets. However, a significant reduction in WG occurred when 600, 800 and 1000 g/kg FM protein was replaced by MBM (p < .05). Similar trends were observed in specific growth rate and protein efficiency ratio. Apparent digestibility coefficients (ADC) of protein and dry matter of the diets S80 and S100 were significantly lower than those of the other diets. The ADC of phosphorus significantly reduced with the increase in dietary MBM level. Nitrogen and phosphorus excretion increased with the increasing dietary MBM level. Protease, lipase and amylase activities of the diets S80 and S100 were significantly lower than those of the other diets (p < .05). The results of this study showed that the optimum dietary MBM replacement level was 34.3% according to broken‐line model based on WG against dietary MBM replacement level.     

Optimum level of dietary monocalcium phosphate determined based on optimal growth and vertebrae phosphate content of juvenile Ussuri catfish,Pseudobagrus ussuriensis

Two experiments were conducted to determine the optimum level of dietary available phosphorus from monocalcium phosphate for juvenile Ussuri catfish Pseudobagrus ussuriensis. Experiment 1 was conducted to estimate phosphorus digestibility from monocalcium phosphate for juvenile Ussuri catfish. The apparent digestibility coefficient of phosphorus from monocalcium phosphate was 86.3%. In the experiment 2, triplicate groups of juvenile Ussuri catfish were fed diets containing graded levels of monocalcium phosphate (MCP: 0 g/kg, 8.2 g/kg, 16.4 g/kg, 24.6 g/kg, 32.8 g/kg and 41.0 g/kg) for 8 weeks. Fish fed the diet containing 16.4 g/kg MCP with available phosphorus of 4.8 g/kg showed the best weight gain (171.5%), feed conversion ratio (1.08) and protein efficiency ratio (2.06). No significant difference was observed in fish survival among the treatments. The best result in terms of phosphorus retention efficiency (46.10%) was observed in fish fed the diet containing 8.2 g/kg MCP with available phosphorus of 3.0 g/kg, which was not different (p > .05) from those fed the diet containing up to 24.6 g/kg MCP, and the highest vertebrae phosphorus content (58.2 g/kg) was observed in fish fed the diet containing 24.6 g/kg MCP with available P of 6.6 g/kg. The whole‐body lipid and protein, as well as phosphorus contents, were significantly affected by dietary available phosphorus (p < .05). Viscerosomatic index (VSI) and condition factor (CF) were inversely correlated with dietary phosphorus levels (p < .05). Quadratic regression analysis based on specific growth rate (SGR) against dietary available phosphorus levels indicated that the optimum available phosphorus requirement for the maximal growth of juvenile Ussuri catfish was 5.9 g/kg, and broken‐line analysis based on vertebrae phosphorus content against dietary available phosphorus levels indicated that a dietary level of 6.0 g/kg available phosphorus will provide optimum vertebrae phosphorus content.     

Effects of fishmeal replacement with cottonseed meal protein hydrolysate on growth,digestion and intestinal histology of juvenile Chinese soft‐shelled turtle,Pelodiscus sinensis

A 60‐day feeding experiment was conducted to evaluate the effects of fishmeal (FM) replacement with cottonseed meal protein hydrolysate (CPH) on growth, digestion and intestinal histology of juvenile Chinese soft‐shelled turtle (Pelodiscus sinensis). Five diets were formulated to replace 0, 50, 100, and 150 g/kg fishmeal protein by CPH (CPH0, CPH5, CPH10, CPH15) and CPH15L (CPH15 with micro capsule‐L‐lysine). Weight gain, feed conversion rate and protein efficiency ratio showed no significant differences compared to control group (p > .05). The highest feed intake indicated in CPH15 (p < .05). The composition of whole‐body varied slightly in each groups (p > .05). The trypsin activity significantly elevated when dietary fishmeal protein was replaced by CPH at 30–90 g/kg (p < .05). A significantly higher lipase activities in CPH5 than control group (p < .05). The CPH5‐10 groups showed higher villus height than the other groups (p < .05). The microvillus length in turtles with CPH showed a significant increasing length (p < .05). The results indicated that replacing up to 90 g/kg of dietary fishmeal protein with CPH did not hamper growth or reduce feed intake of turtles. Moreover, CPH replaced 60 g/kg FMP can increase intestine digestive enzymes activities and improve intestinal development.     

Effect of dietary leucine on growth performance,hemolymph and hepatopancreas enzyme activities of swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to determine the dietary leucine requirement for juvenile swimming crabs reared in cement pools. Six isonitrogenous and isolipidic practical diets (430 g/kg crude protein and 70 g/kg crude lipid) were formulated to contain graded leucine levels which ranged from 16.7 to 26.7 g/kg (dry weight). Each diet was randomly assigned to triplicate groups of 60 juvenile swimming crabs (initial average weight 3.75 ± 0.12 g) that were stocked in rectangle plastic baskets. The results of the present study indicated that dietary leucine levels significantly influenced weight gain (WG) and specific growth ratio (SGR) (p < .05), crab fed the diet containing 22.7 g/kg leucine had significantly higher WG and SGR than those fed the other diets. Feed efficiency and protein efficiency ratio were not significantly affected by the dietary leucine levels (p > .05). Total protein, cholesterol, triglyceride and glucose in serum were significantly affected by the dietary leucine levels. Aspartate aminotransferase (AST) and alanine aminotransferase activities in hemolymph, AST and superoxide dismutase activities in hepatopancreas were significantly affected by dietary leucine levels; moreover, crab fed the 16.7 g/kg leucine diet had higher malondialdehyde in hemolymph and hepatopancreas than those fed the other diets. Crab fed the diet containing 24.9 g/kg leucine had higher phenoloxidase activity in hemolymph than those fed the other diets. Based on two‐slope broken‐line model of SGR against dietary leucine levels, the optimal dietary leucine requirement for growth was estimated to be 22.1 g/kg of the dry diet (corresponding to 51.4 g/kg of dietary protein on a dry weight basis). In summary, findings of this study indicated that dietary leucine could improve growth performance and antioxidant status.     

Effects of Chinese herbal medicines mixture on growth performance digestive enzyme activity immune response of juvenile Japanese seabass,Lateolabrax japonicus

A single factorial experiment was conducted to investigate the effects of Chinese herbal medicines mixture (CHMM) on growth performance, digestive enzyme activity and immune response of Japanese seabass, Lateolabrax japonicus (initial weight 5.01 ± 0.32 g). The fish were fed diets containing six levels of CHMM (0, 4, 8, 12, 16 and 20 g/kg) for 4 weeks. The results showed that the weight gain rate and specific growth rate (SGR) enhanced significantly in fish fed diet containing 8 g/kg CHMM (p < .05), while the feed conversion ratio (FCR) in the 4 and 8 g/kg CHMM groups reduced significantly compared with the control (p < .05). The body crude protein levels in 8 and 16 g/kg groups were significantly higher than the control (p < .05). The pepsin, amylase and lipase activities elevated significantly in the stomach of fish fed 8 g/kg CHMM, while the erepsin and lipase in the intestine of fish in 12 and 16 g/kg groups were higher significantly than the control (p < .05). The lysozyme, alkaline phosphatase, acid phosphatase (ACP), total antioxidative capacity activities in serum of fish fed 12 g/kg CHMM were higher significantly than those in the control (p < .05), while the total superoxide dismutase (SOD), total antioxidative capacity, catalase, alkaline phosphatase (AKP) and ACP activities in hepatopancreas of fish in 12 g/kg group were all significantly higher than those in the control (p < .05). Regression analysis showed that the relationships between dietary CHMM levels and either FCR, SGR, erepsin, pepsin or lysozyme activities were best expressed by quadratic or cubic regression equations, and the optimal inclusion levels are 11.4, 10.7, 10.7, 8.4 and 10.5 g/kg for maximum FCR, SGR, erepsin, pepsin and lysozyme activities, respectively. Under the present experimental condition, the optimal supplementary level of CHMM in the diet of Japanese seabass is 8–12 g/kg.     

Effects of xylan and cottonseed meal on growth performance,digestion, intestinal morphology and the expression of two peritrophin genes of Chinese mitten crab,Eriocheir sinensis

Xylan is one of the main non‐starch polysaccharides (NSPs) in cottonseed meal. This study aimed to evaluate the anti‐nutrition effect of xylan and to better understand cottonseed meal as a feed ingredient for the Chinese mitten crab, Eriocheir sinensis. A diet containing 280 g/kg fishmeal without cottonseed meal or xylan was formulated as a control (FM diet). A second diet (XYL diet) was supplemented with 53.3 g/kg xylan according to its amount in the third diet, the cottonseed meal diet (CSM diet), which was formulated to include 400 g/kg cottonseed meal. The last diet (CSM + XYLase diet) included 400 g/kg xylanase hydrolysed cottonseed meal, in which most xylan was hydrolysed. Diets were isonitrogenous (370 g/kg crude protein) and isoenergetic (17.58 KJ/g) and were randomly fed to 12 tanks of crabs for 8 weeks. Results showed that no difference was detected among groups in growth performance, serum biochemical index, or digestive and metabolism enzyme activity (p > 0.05). The apparent digestibility coefficient (ADC) of dry matter was FM > XYL > CSM + XYLase > CSM (p < 0.05). The ADC of protein in crab fed diet FM and XYL was significantly higher than CSM and CSM + XYLase (p < 0.05). Cottonseed meal decreased the fold height of the hindgut, upregulated the expression of peritrophin genes of Es‐PL44 and Es‐PP1 (p < 0.05), while xylan decreased the fold height of the hindgut and upregulated the expression of Es‐PP1 (p < 0.05). These results indicated that the negative effect of cottonseed meal and xylan on digestibility partly compensated by enhanced absorption of the hindgut innerly and by supplementation of xylanase externally, and cottonseed meal is an attractive alternative protein source in general for Chinese mitten crab.     

Effect of dietary carbohydrate levels on growth performance,non‐specific immune enzymes and acute response to low salinity and high temperature of juvenile sea cucumber Apostichopus japonicus

This study was performed to determine the optimum dietary carbohydrate (CHO) levels of sea cucumber, based on the parameters of growth, digestive enzymes, digestibility, non‐specific immune enzymes and acute low‐salinity (20 g/L) stress and high‐temperature (30°C) stress tolerance. Diets with eight different CHO (dextrin) levels (32.9, 107.6, 192.5, 257.2, 316.8, 428.0, 482.4 and 572.8 g/kg) were fed to sea cucumber juveniles (0.49 ± 0.01 g) for 60 days. Significant higher amylase activity was observed in sea cucumbers fed diet with CHO ranging between 32.9 and 192.5 g/kg than that of other treatments (p < .05). The sea cucumbers fed with 192.5 g/kg CHO showed significantly higher acid phosphatase activity than the treatments of 482.4 and 572.8 g/kg CHO (p < .05), and significantly higher alkaline phosphatase activity than other treatments (p < .05, except 257.2 g/kg). The treatments of 428.0–572.8 g/kg were found significantly lower values than other treatments in apparent digestibility coefficients for dry matter and crude protein (p < .05). The sea cucumbers fed with 192.5, 257.2 and 316.8 g/kg CHO showed better tolerance to high‐temperature (30°C) and low‐salinity (20 g/L) stress than other treatments. In brief, the optimal dietary CHO level for the growth of juvenile sea cucumber is 177.96 g/kg. However, excessive CHO will inhibit amylase enzyme activity and decrease digestibility, resulting in low growth of sea cucumber.     

Dietary protein/carbohydrate ratio in low‐lipid diets for Senegalese sole (Solea senegalensis,Kaup 1858) juveniles. Influence on growth performance,nutrient utilization and flesh quality

Four isoenergetic (21 kJ/g dry matter, DM) and isolipidic (65 g/kg DM) diets containing different crude protein/total carbohydrate (CHO) ratios: 60/26, 56/30, 52/34 and 48/38, were tested in 22 g Senegalese sole for 104 days. Apparent digestibility coefficients (ADC) were not affected by the treatments, and all groups presented extremely low starch ADC values (22.8%–36.5%). Replacement of dietary protein by CHO did not affect daily growth index (0.9–1.0), but significantly increased voluntary feed intake of fish. Regression analyses demonstrated that digestible protein content, rather than digestible energy, was the main dietary factor influencing such feeding activity (R² = .952). A significantly increased feed conversion ratio was observed in sole fed increasing CHO contents. The dietary protein/CHO ratio did not influence whole‐body composition. Sole fed the 48/38 diet showed the lowest efficiency in terms of N and energy utilization. PUFA were the most represented fatty acid fraction in fillet, regardless of the dietary protein/CHO ratio, mainly due to the high content of DHA. Senegalese sole increase feed intake under low dietary protein/CHO ratios to ensure an adequate N intake. Such compensatory mechanism seems to be triggered to satisfy a specific protein metabolic requirement for energy purposes as tissue accretion remained unchanged.     

Fishmeal levels can be successfully reduced in white shrimp (Litopenaeus vannamei) if supplemented with DL‐Methionine (DL‐Met) or DL‐Methionyl‐DL‐Methionine (Met‐Met)

The main objective of this study was to evaluate the effect of methionine supplementation when reducing fishmeal levels in diets for white shrimp (Litopenaeus vannamei). Tested diets consisted of a positive control with 260 g/kg fishmeal (D1), two negative controls with 100 g/kg fishmeal and no amino acid (AA) supplementation (D2) or supplemented with lysine but not methionine (D3), and four additional diets with 100g/kg fishmeal supplemented with increasing levels of DL‐Met (1.0, 2.0 or 3.0 g/kg) (D4, D5, D6) or Met‐Met (1.0 g/kg) (D7). Each diet was fed to four groups of 30 shrimp for 8 weeks at a daily rate of 70 g/kg body weight. Reduction in fishmeal from 260 g/kg down to 100 g/kg did not significantly affect survival rate, feed conversion ratio (FCR), protein efficiency ratio (PER) or protein retention efficiency (PR%) of white shrimp. However, growth performance (final body weight, FBW; weight gain, WG; specific growth rate, SGR) was reduced when dietary fishmeal level was reduced from 260 g/kg (D1) to 100 g/kg without methionine supplementation (D2). The growth performance (FBW, WG and SGR) of shrimp was significantly increased by supplementation of the 100 g/kg fishmeal diet with increasing levels of DL‐Met (p < .05). Same performance as positive control (D1) was achieved with diets containing 100 g/kg fishmeal and supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. The highest values of growth performance (FBW, WG and SGR) were found in shrimp fed D6 and D7 diets, which were significantly higher than those of shrimp fed D2 and D3 diets (p < .05) but without statistical differences with shrimp fed D1, D4 and D5 diets (p > .05). The highest values of whole‐body and muscle protein contents were found in shrimp fed D1 diet, which were significantly higher than those of shrimp fed all other diets (p < .05). The highest value of intestinal tract proteolytic enzyme activity was found in shrimp fed Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) and 3 g/kg DL‐Met‐supplemented diet (D6) (p < .05). The highest values of apparent digestibility coefficients (ADCs) of dry matter and crude protein were found in Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) (p < .05). Shrimp fed the D1 diet showed the highest value of total essential amino acid (EAA) and was significantly higher than shrimp fed D2–D3 (p < .05) but without significant difference with shrimp fed D4–D7 (p > .05). In conclusion, results showed that same performance can be achieved with diets containing 260 or 100 g/kg fishmeal supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. Moreover, supplementation of limiting methionine in low‐fishmeal diets seems to improve the digestive proteolytic activity, improving digestibility of dry matter and protein, and eventually to promote growth of juvenile white shrimp in fishmeal reduction diets.