Dietary inorganic phosphorus or microbial phytase supplementation improves growth,nutrient utilization and phosphorus mineralization of juvenile red sea bream,Pagrus major,fed soybean‐based diets

A 2 × 3 factorial design with triplicates examined the interaction between dietary inorganic phosphorus (IP) and phytase on growth, mineral utilization and phosphorus (P) mineralization in juvenile red sea bream. The treatments were three levels of dietary IP supplementation at 0, 2.5 and 5 g kg^?1, either without or with phytase supplementation [2000 FTU kg^?1; phytase unit is defined as the amount of enzyme activity which liberates 1 micromol of inorganic phosphorus per minute at pH 5.5 and 37 °C at a substrate concentration (sodium phytate) of 5.1 mmol L^?1]. Juvenile red sea bream (IBW = 1.3 g ± 0.1) were stocked twelve fish per tank and fed for 50 days. Growth and feed efficiency were significantly (P < 0.05) enhanced by both dietary P and phytase supplementation. Feed intake and survival rate were not significantly affected by the dietary treatments. Both dietary IP and phytase supplementation significantly increased plasma IP and Mg levels. Concentration of vertebral mineral and scale P was significantly increased by both dietary treatments. A skeletal malformation syndrome of scoliosis occurred in fish fed both non‐IP and non‐phytase supplemented diet. Interaction between main dietary effects was detected for vertebral Zn, scale P and whole‐body ash and Mg content. With regard to growth and other examined productivity traits, phosphorus requirement of juvenile red sea bream can be met if supplemented with 2000 FTU phytase kg^?1 or in the absence of phytase, by dietary inclusion of 2.5–5 g kg^?1 of IP.     

Comparative study on the effect of high dietary carbohydrate on the growth performance,body composition,serum physiological responses and hepatic antioxidant abilities in Wuchang bream (Megalobrama amblycephala) and black carp (Mylopharyngodon piceus Richardson, 1846)

A 90‐day trial investigated the effect of dietary carbohydrate level on growth performance, body composition, serum physiological responses and hepatic antioxidant abilities of a herbivorous fish, Wuchang bream (Megalobrama amblycephala) and a carnivorous fish, black carp (Mylopharyngodon piceus Richardson, 1846). For each species, two isonitrogenous and isolipidic semi‐purified diets (28.6% crude protein and 5.3% crude lipid for Wuchang bream) were formulated to contain 30.4% and 52.9% carbohydrate (WB‐NCHO and WB‐HCHO); Two isonitrogenous and isolipidic semi‐purified diets (32.6% crude protein and 5.3% crude lipid for black carp) were formulated to contain 20.5% and 40.5% carbohydrate (BC‐NCHO and BC‐HCHO). It was shown that, compared to Group WB‐NCHO, Wuchang bream in Group WB‐HCHO had lower weight gain rate, specific growth rate, serum total protein, hepatic superoxide dismutase activity and total antioxidative capacity, while higher feed conversion ratio, serum triglyceride and cortisol levels (P < 0.05). Compared to Group BC‐NCHO, black carp in Group BC‐HCHO had lower final weight, weight gain rate, specific growth rate, protein efficiency ratio, serum alkaline alkaline phosphatase activity, serum total protein, hepatic superoxide dismutase activity and total antioxidative capacity, while higher feed conversion ratio, whole body crude lipid, serum triglyceride, cholesterol, glutamic‐oxaloacetic transaminase activity and cortisol levels (P < 0.05). The results of this study suggested that Wuchang bream had better capacity of utilizing dietary carbohydrate than that of black carp, while high levels of dietary carbohydrate might not only be detrimental to growth performance and feed utilization but also the immune response and health for both species.     

Effects of replacing fishmeal with fermented and non‐fermented rapeseed meal on the growth,immune and antioxidant responses of red sea bream (Pagrus major)

The effect of rapeseed meal (RM) and Aspergillus oryzae fermented rapeseed meal (RM‐Koji) on red sea bream (Pagrus major) was examined. Three groups of fish (initial weight, 4.5 ± 0.02 g) were fed a basal diet (RM0) and two test diets where half of fishmeal was replaced by RM (RM50) and RM‐Koji (FRM50) for 56 days. The obtained results showed that fish fed RM0 and FRM50 exerted significantly higher growth performance, feed utilization and haemoglobin level but lower triglyceride and cholesterol than RM50 group (p < 0.05). Interestingly, except of antiprotease activity, all the immune parameters including lysozyme, respiratory burst (NBT) and bactericidal activities were significantly increased in fish fed RM0 and FRM50 diets compared to RM50 diet (p < 0.05). In addition, malondialdehyde and reactive oxygen metabolites were significantly reduced in RM0 and FRM50 groups over RM50 group (p < 0.05). The present results suggest that fermented RM induced better growth performance and immune responses than feeding red sea bream with non‐fermented RM and both RM and RM‐Koji improved the antioxidative status of fish, making RM‐Koji an interesting candidate as a functional feed for aquatic animals.     

Evaluation of pea proteins and poultry protein as fish meal alternatives in the diets for juvenile black sea bream,Acanthopagrus schlegelii

The potential of three different protein resources (pea protein isolate, PPI; pea protein concentrate, PPC; enzyme treated poultry protein, ETPP) as fish meal (FM) alternative protein in diets for juvenile black sea bream, Acanthopagrus schlegelii. (initial average weight 7.90 ± 0.13 g) was evaluated. Seven isonitrogenous and isoenergetic diets were formulated to replace FM at 0% (T0, control diet), 8% (designated as T1‐T3) and 16% (designated as T4‐T6) using PPI, PPC and ETPP respectively. Each diet was randomly assigned to triplicate groups of 25 juvenile fish for 8 weeks. At the end of the feeding period, survival rate was not significantly affected by dietary treatments. Growth performance in T6 (16% ETPP) group was significantly inferior to T0 group, however, weight gain and specific growth rate in other treatments showed no significant differences (P > 0.05). Mean feed intake, feed efficiency ratio and protein efficiency ratio were also poorer in fish fed in T6 than those of fish fed with the control diet respectively. Apparent digestibility coefficients (ADCs) of dry matter and crude protein for fish fed ETPP diets were significant lower than those of fish fed with the control diet, whereas ADCs of lipid were unaffected by dietary treatments. ADC's of dietary Leu, Ile, His and Lys was also significantly influenced. There were no marked variations in proximate compositions of dorsal muscle. With regard to plasma characteristics, significant difference was observed in triacylglycerol content. Ammonia concentration in plasma tended to increase in alternative protein diets as substitution level increased. There were significant differences in aspartate aminotransferase activities among groups, but alanine aminotransferase levels were unaffected by treatments. In conclusion, the present study demonstrated that PPI and PPC were potential protein sources for using in juvenile black sea bream diet. However, the substitution level of FM by ETPP should be limited within 16%.     

Effects of dietary 60 g kg−1 dried distiller's grains in least‐cost practical diets on production and gut allochthonous bacterial composition of cage‐cultured fish: comparison among fish species with different natural food habits

Dried distiller's grain (DDG) is considered as an alternative ingredient of dietary feed due to its high contents of protein, fibre and fat. In this study, 60 g kg^‐1 of DDG was used to feed grass carp (Ctenopharyngodon idella), bluntnose black bream (Megalobrama amblycephala), gibel carp (Carassius gibelio) and black carp (Mylopharyngodon piceus) for 8 weeks, and its effect on fish production and gut allochthonous microbiota was investigated for the development of a suitable fish feed high in nutrients and low in cost for polyculture freshwater fish. DDG supplementation resulted in the less weight gain and higher feed conversion ratio of black carp (P < 0.05), but had no significant effects on other fish or parameters. PCR–denaturing gradient gel electrophoresis (DGGE) analysis indicated that all four fish species had some common and unique bacteria in their digestive tracts, and the gut microbiota of bluntnose black and gibel carp fed the control diet and DDG diets were very similar (Cs > 91%); of them, the total counts of intestinal bacteria studied by qPCR increased in grass carp (P < 0.05) and depressed in black carp (P < 0.05) when fed dietary DDG. Thus, we assumed that dietary DDG modulated production and gut microbiota of fish in a host‐specific way.     

Effects of dietary taurine levels on epidermal thickness and scale loss in red sea bream,Pagrus major

Taurine is often added to artificial fish diets to compensate for a reduction in fish meal (FM). However, the taurine content of FM‐based diets is typically lower than in diets consisting of raw fish, even in diets where FM is the only protein source. We evaluated the effects of dietary taurine in FM‐based diets on epidermal thickness and scale detachability in red sea bream Pagrus major. We compared the effect of diets containing 0% (control), 0.3% (Tau‐0.3%), 0.6% (Tau‐0.6%) and 1.0% (Tau‐1.0%) taurine. Red sea bream (average body weight, 39 g) were fed these diets for 7 weeks. Taurine supplementation had no effect on growth, feed intake, feeding efficiency, or survival. However, the epidermal thickness was higher in fish in the Tau‐0.6% and Tau‐1.0% groups than in the control and Tau‐0.3% groups. Similarly, scale loss was significantly higher in the control group than in the Tau‐0.6% and Tau‐1.0% groups. Our results suggest that supplementation with >0.6% taurine (1.0% in diet) improves skin condition.     

Effect of dietary replacement of fish meal by soybean protein concentrate on growth performance and phosphorus discharging of juvenile black sea bream,Acanthopagrus schlegelii

The 8‐week experiment was conducted to evaluate the effects of partial replacement of fish meal (FM) with soybean protein concentrate (SPC) on juvenile black sea bream, Acanthopagrus schlegelii (10.70 ± 0.04 g). Diets were formulated to replace FM protein by SPC at 0, 8, 16, 24, 32 or 40% (designated as T1, T2, T3, T4, T5 and T6, respectively). Diets except T1 were supplemented with phytase at 2000 phytase activity U kg^?1. The results showed that survival rate, growth performance and feed utilization were not significantly affected by increasing dietary SPC. Fish fed diet T3 had higher feed intake compared to those fed T1, T2 and T5 diets. Whole body compositions of black sea bream were significantly influenced by SPC replacing FM except for protein, ash and phosphorus content. Condition factor of fish was significantly lower in T2 than that of fish in T3 group. Apparent digestibility coefficients (ADCs) of dry matter was higher in fish fed T6 diet than those of fish fed T1 and T2 diets, ADCs of phosphorus increased with dietary SPC level up to T3 and then decreased. The results obtained in this study indicate that FM protein could be effectively replaced by SPC protein with phytase in diet of black sea bream.     

Effects of Dietary Vitamin C on Blood Chemistry and Nonspecific Immune Response of Juvenile Red Sea Bream,Pagrus major

A trial was conducted to determine the effect of ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) on blood chemistry and nonspecific immune response of red sea bream juveniles. Test diets with three levels of AsA (free, 107, and 325 mg/kg diet) were fed to juvenile red sea bream (36.0 ± 1.3 g) two times a day for 3 wk. There were no significant differences in hematocrit, glucose, and blood urea nitrogen. Total cholesterol and triglyceride in plasma of fish fed AsA‐free diet was significantly (P < 0.05) higher than that of fish fed two other diets. There were no significant differences in serum albumin, total bilirubin, and total serum protein. Glutamyl oxaloacetic transaminase in serum of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) lower than that of fish fed AsA‐free diet. Serum lysozyme activity (LA) of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) higher than that of fish fed AsA‐free diet. There was no significant difference in mucus LA. The results mentioned above demonstrated that AMP‐Na/Ca is a bioavailable AsA source for red sea bream juveniles. Supplement of more than 107 mg AsA/kg in diets improved blood chemistry and nonspecific immune function of red sea bream juveniles.     

Effects of different dietary lipid sources on tissue fatty acid composition,serum biochemical parameters and fatty acid synthase of juvenile mud crab Scylla paramamosain (Estampador 1949)

A 6‐week study was conducted to determine the effects of different lipid sources in pelleted diets on juvenile mud crab Scylla paramamosain. Five isonitrogenous and isolipidic diets containing 8% level of fish oil (FO), lard (LD), safflower oil (SO), perilla seed oil (PO) or mixture oil (MO; V_FO:V_SO:V_PO = 1:1:1), and a live food of marine bivalve Potamocorbula rubromuscula as the control diet (CF), were fed to groups of 25 juvenile crabs (average initial weight 7.4 g, carapace width 3.5 cm) in triplicate. The results showed that crabs fed MO had the highest survival (P < 0.05). The moisture content was significantly higher in crabs fed LD, SO and PO (P < 0.05). Crabs fed SO exhibited the lowest crude protein and lipid (P < 0.05). Ash contents were obviously lower in LD group (P < 0.05). Highest total lipid in the hepatopancreas and muscle was in LD and FO group respectively. Glucose, total cholesterol and low‐density lipoprotein were higher while high‐density lipoprotein was lower (P < 0.05) in LD group. Tissue fatty acid compositions were consistent with those in diets. FO and MO diets had the same depression effect like CF on fatty acid synthase activity and mRNA expression in the hepatopancreas. The results of this study indicated that FO and mixed oil are suitable for preparation of pelleted diets with better effects for juvenile S. paramamosain compared with live food, and the ratio of n‐6/n‐3 fatty acids in pelleted diets must be <1.     

A natural infection by the red sea bream iridovirus‐type Megalocytivirus in the golden mandarin fish Siniperca scherzeri

An outbreak of a Megalocytivirus infection was found in the golden mandarin fish Siniperca scherzeri during September and October 2016, in Korea. Phylogeny and genetic diversity based on the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes showed a new strain. Designated as GMIV, this strain derived from the golden mandarin fish was suggested to belong to the red sea bream iridovirus (RSIV)‐subgroup I. Additionally, this train clustered with the ehime‐1 strain from red sea bream Pagrus major in Japan and was distinguished from circulating isolates (RSIV‐type subgroup II and turbot reddish body iridovirus [TRBIV] type) in Korea. The infection level, evaluated by qPCR, ranged from 8.18 × 10² to 7.95 × 10⁶ copies/mg of tissue individually, suggesting that the infected fish were in the disease‐transmitting stage. The diseased fish showed degenerative changes associated with cytomegaly in the spleen as general sign of Megalocytivirus infection. The results confirm that the RSIV‐type Megalocytivirus might have crossed the environmental and species barriers to cause widespread infection in freshwater fish.     

Evaluation of Schizochytrium meal in microdiets of Pacific white shrimp (Litopenaeus vannamei) larvae

A feeding trial was conducted to assess the effects of dietary Schizochytrium meal supplementation on survival, growth performance, activities of digestive enzymes and fatty acid composition in Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Four isonitrogenous and isolipidic diets were formulated to contain graded levels of Schizochytrium meal: 0% (S0, the control diet), 2% (S2), 4% (S4) and 6% dry matter (S6). Results showed that there was no significant difference in survival of shrimps among dietary treatments (P > 0.05). Shrimps fed diets with 2% and 4% microalgae meal had significantly higher specific growth rate (SGR) than that of shrimps fed diets with 0% and 6% microalgae meal, and no significant differences were observed between shrimps fed diets with 2% and 4% microalgae meal (P > 0.05). Activity of trypsin in the pancreatic and intestinal segments, and activity of amylase in the pancreatic segments were not significantly affected by dietary microalgae meal levels (P > 0.05). Specific activities of both alkaline phosphatase and leucine‐aminopeptidase in intestine and purified brush border membrane of intestine were significantly higher in shrimps fed diet with 2% microalgae meal (P < 0.05). There were no significant differences in C18:2n‐6, n‐3 fatty acids, n‐6fatty acids, PUFA and n‐3/n‐6 in muscle samples among dietary treatments. C16:1n‐7, C18:1n‐9, MUFA, C18:3n‐3 and C20:5n‐3 decreased, however, C20:4n‐6 increased in the muscle as dietary microalgae meal level increased. In conclusion, 4% Schizochytrium meal in microdiets of shrimps can improve growth performance and may be a valuable additive in the microdiets of shrimps.     

Effects of Fungal Phytase on Utilization of Dietary Protein and Minerals, and Dephosphorylation of Phytic Acid in the Alimentary Tract of Channel Catfish Ictalurus punctatus Fed an All-Plant-Protein Diet

A feeding trial was conducted to quantify the effects of phytase at levels of 0, 500, 1,000, 2,000, 4,000, and 8,000 units (U) per kg diet on utilization of dietary protein and minerals by fingerling (12 g) channel catfish Ictalurus punctatus fed an all‐plant‐protein diet composed of soybean meal, corn, and wheat middlings. The effects of phytase on dephosphorylation of phytic acid (phytate) in the alimentary tract of catfish also were determined. After 14 wk, mean weight gains (30.2–43.9 g/fish), feed conversion ratios (2.27–2.40 g feed consumed/g weight gain), protein efficiency ratios (1.47–1.61 g weight gaid/g protein consumed), and dietary protein retentions (23.8–26.7%) did not differ significantly (P > 0.05) among treatment groups. A digestibility trial conducted after the feeding trial showed no difference (P > 0.05) in mean digestibility of diet dry matter (49.0–58.3%) or crude protein (85.4‐88.5%) among treatment groups. Concentrations of ash (46.7–48.6%), calcium (Ca, 17.9–18.5%), phosphorus (P, 9.1–9.5%), and manganese (Mn, 65.5–74.1 mg/kg) were significantly higher (P ≤ 0.05) in bone of fish fed ≥ 500 U/kg than in bone of fish fed 0 U/kg (ash, 43.5%; Ca, 16.4%; P, 8.4%; and Mn, 49.0 ma/kg), but concentrations of these minerals did not differ (P > 0.05) in bone of fish fed ≥ 500 Uk/g. The magnesium (Mg) content of bone did not differ (P > 0.05) between fish fed 0 U/kg (0.29%) or 500 U/kg (0.34%), but was significantly lower in fish fed 0 U/kg than in fish fed ≥ 1,000 U/kg (0.35–37%). Bone Mg levels did not differ (P > 0.05) among fish fed ≥ 500 U/kg. The amount of zinc (Zn) in bone of fish fed 8,000 U/kg (153.3 mg/kg) was significantly higher than that in fish fed 0 U/kg (115.7 mg/kg) or 500 U/kg (130.3 mg/ kg), but did not differ from Zn levels in bone of fish fed 1,000–4,000 U/kg (134.5–135.8 mg/ kg). Dephosphorylation of phytate occurred primarily in the stomach within 2–8 h after diet ingestion, depending on the level of phytase supplementation. Initial levels of total phytate in the diet decreased 32–94% in stomach contents of fish fed l,000–8,000 U/kg within 2 h after feeding. Eight hours after feeding, stomach contents of fish fed ≥ 1,000 U/kg contained less than 6% of initial total dietary phytate. Stomach contents of fish fed 500 U/kg retained 92% of initial total dietary phytate 2 h after feeding and 15% of total dietary phytate 8 h after feeding. Results of this study indicate that phytase supplementation at levels up to 8,000 U/kg diet did not increase weight gain or improve dietary protein utilization of channel catfish fed an all‐plant‐protein diet. Addition of phytase at a level of 1,000 U/kg diet was sufficient to significantly increase the Ca, P, Mg, and Mn content of bone, relative to fish fed an unsupplemented diet, and significantly decrease the quantity of total phytate in feces. A phytase level of 8,000 U/kg diet significantly increased the bioavailability of naturally occurring Zn in feed ingredients and increased the rate of phytate dephosphorylation in the stomach, compared with a diet containing no added phytase. Increased utilization of naturally occurring minerals in feed ingredients reduces the need for mineral supplements in diets and results in decreased elimination of minerals in feces. Thus, use of phytase in catfish feeds can be expected to provide both economic and environmental benefits.     

Prebiotics effect on growth performance,hepatic intermediary metabolism,gut microbiota and digestive enzymes of white sea bream (Diplodus sargus)

This study aimed at evaluating the effects of short‐chain fructooligosaccharides (scFOS), xylooligosaccharides (XOS) and galactooligosaccharides (GOS) on growth performance, hepatic metabolism, gut microbiota and digestive enzymes activities of white sea bream juveniles. Four diets were formulated: a control diet with fish meal (FM) and plant feedstuffs (PF) (30FM:70PF) and three test diets similar to control but supplemented with 10 g of scFOS, XOS or GOS per kilo diet, which were fed to fish during 12 weeks. Prebiotics had no effect on growth, feed efficiency or gut microbiota. Plasmatic triglycerides were lower in fish fed XOS than FOS and GOS diets. Malic enzyme activity was lower in fish fed XOS than FOS diet. Fish fed XOS diet had lower fatty acid synthetase (FAS), a key lipogenic enzyme and higher alanine aminotransferase activities. Fifteen days after the start of the trial, an enhancement of total alkaline protease, trypsin and lipase activities was observed in fish fed prebiotics, but such effect disappeared at 12 weeks. In conclusion, scFOS, XOS or GOS seem to have limited applicability in white sea bream feed.     

Effect of dietary Yucca schidigera extract on growth,total ammonia–nitrogen excretion and haematological parameters of juvenile striped catfish Pangasianodon hypophthalmus

A feeding trial with striped catfish, Pangasianodon hypophthalmus was performed to determine the effect of Yucca schidigera in practical diet on the growth, feed utilization, body composition, total ammonia–nitrogen (TAN) excretion and haematological parameters. A diet with fish meal as the main protein source without yucca extract was used as the control diet (Diet 1). Four diets were formulated with 0.075 (Diet 2), 0.1 (Diet 3) and 0.15 (Diet 4)% of yucca extract respectively. Fifteen fish per tank (initial mean weight 1.78 ± 0.05 g) were randomly allocated to 15 fibreglass tanks (80‐L) connected to a freshwater closed recirculation system (temperature 29.7 ± 1.0°C). The experimental diets were tested in triplicates for 12 weeks. The specific growth rate of fish fed Diet 4 was significantly higher when compared with fish fed Diet 1. The growth of fish fed diets Diet 2 and Diet 3 were not significantly different compared with fish fed the Diet 1. Striped catfish fed Diet 4 had significantly lower feed conversion ratio compared with fish fed Diet 1 and Diet 2 (P < 0.05).The incorporation of high level Yucca schidigera extract in the diets reduced TAN compared with Diet 1. Dietary inclusion of Yucca extract levels did not significantly affect the biometric parameters or whole body proximate composition of the striped catfish (P > 0.05). The PCV (%) in fish significantly increased with high levels of Yucca inclusion (Diet 4) compared with control diet. Fish fed Diet 4 showed significantly higher haemoglobin levels than Diet 1 (P < 0.05). The results indicate that dietary inclusion of Yucca schidigera extract is promising as a feed additive that could improve growth performance and some haematological parameters and the best Yucca schidigera level was 0.15%.     

Soy protein concentrate as a substitute for fish meal in diets for juvenile Acanthopagrus schlegelii: effects on growth,phosphorus discharge and digestive enzyme activity

The current high demand and cost of fish meal (FM) necessities the evaluation of alternative plant protein ingredients in diets of farmed marine fish. A 56‐day feeding trial was performed to study the effects of replacement of FM with soy protein concentrate (SPC) in diets of Acanthopagrus schlegelii. Diets were prepared at levels of 0%, 30%, 47.5%, 65%, 82.5% and 100% SPC, respectively, replacing FM. The results indicated no significant differences (p > .05) in % weight gain (WG) and specific growth rate of fish fed S30% to S82.5% diets compared with the control diet while, further inclusion at 100% significantly depressed growth performance. SPC inclusion and phytase supplementation significantly affected the phosphorous discharge (P‐load) showing lowest value (3.83 ± 0.53 g/kg WG) in S100% compared to control (14.79 ± 0.37 g/kg WG) and in fish fed S30% diet (13.24 ± 0.89 g/kg WG) (p < .05). The results of this study showed that FM could be substituted up to 82.5% by SPC in the diet of Acanthopagrus schlegelii fingerlings (5.53 ± 0.12 g) without any adverse effects. Phytase supplementation SPC based diets could be effective in reducing the phosphorus load in the aquatic culture environment.     

Tributyrin‐supplemented high‐soya bean meal diets of juvenile black sea bream,Acanthopagrus schlegelii: Study on growth performance and intestinal morphology and structure

An 8‐week growth trial was conducted to evaluate the effects of different levels of tributyrin supplementation in a high‐soya bean meal diet on juvenile black sea bream (11.30 ± 0.16 g). The positive control (PC) diet contained 45% fishmeal and 20% soya bean meal, while the negative control (NC) contained 12% fishmeal and 45% soya bean meal. Graded levels of tributyrin were added to the NC diet at 0.05% (TB 0.05), 0.1% (TB 0.1), 0.2% (TB 0.2), 0.4% (TB 0.4) and 0.8% (TB 0.8). Ultimately, the fish fed the PC diet had a higher weight gain and specific growth rate than the fish fed other diets. The fish fed the NC diet had the lowest growth, and TB 0.05–TB 0.2 diets increased growth performance while TB 0.4–TB 0.8 diets caused reduction in growth. Dietary tributyrin supplementation improved protease activity and enhanced antioxidant capacity. Compared with the fish fed the NC diet, the fish fed the tributyrin‐supplemented diets had improved gut morphology and structure, and the results were similar to those of the fish fed the PC diet. Furthermore, the analysis of the dose response with second‐order polynomial regression indicated that the optimum tributyrin supplementation for juvenile black sea bream is 2.24 g/kg in the 45% soya bean meal diet.     

Effects of dietary n‐3 to n‐6 fatty acid ratios on spawning performance and larval quality in tongue sole Cynoglossus semilaevis

This study investigated the effect of n‐3 to n‐6 fatty acid ratios in broodstock diets on reproduction performance, fatty acid composition of eggs and gonads of tongue sole Cynoglossus semilaevis. Broodstock were fed five isonitrogenous and isoenergetic diets for 60 days. The supplemented lipids were prepared by a combination of fish oil and soybean oil inclusion FO (fish oil); FSO1 (fish oil: soybean oil = 7:1); FSO2 (fish oil: soybean oil = 2.2:1); FSO3 (fish oil: soybean oil = 1:1); FSO4 (fish oil: soybean oil = 1:4.3) as lipid sources with different n‐3 to n‐6 fatty acid ratios 10.40, 5.21, 2.81, 1.71 and 0.87. Results showed that relative fecundity, fertilization rate and survival rate of larvae at 7 days posthatching were all higher in broodstock fed FSO1 and FSO2 diet and significantly (P < 0.05) decreased in groups fed FSO3 and FSO4 diets. The best result in starvation tolerance test was obtained in FSO2 diet. The present study suggests that n‐3 and n‐6 PUFA ratio in broodstock diet has a considerable effect on spawning performance, egg and larval quality for C. semilaevis.     

Effect of guava Psidium guajava (L.) aqueous extract diet on growth performance,intestinal morphology,immune response and survival of Oreochromis niloticus challenged with Aeromonas hydrophila

An 84‐day feeding trial was carried out to evaluate the effects of aqueous Psidium guajava leaf extract (PGE) on growth, intestinal morphology, physiology, immune response and susceptibility of Oreochromis niloticus to Aeromonas hydrophila. Diets containing 0% (P0), 0.25% (P1), 0.50% (P2), 0.75% (P3) and 1.00% (P4) PGE were fed to triplicate groups of fish (mean weight; 1.32 ± 0.04 g) for 84 days. After the 84‐day feeding trial, test fish were injected with pathogenic A. hydrophila and then fed for 14 days. More feed were consumed in groups of fish fed PGE‐treated diets and resulted in significantly higher weight gain and feed intake. Incidentally, there was an increase in the calculated area of absorption of fish fed PGE diets, as accounted for by marginally higher villi length and width. Antioxidant and immune response were improved with PGE inclusion in diets as total protein, superoxide dismutase, glutathione peroxidase and glutathione S‐transferase significantly increased (p < 0.05) in fish fed PGE diets. Results of the challenge test with A. hydrophila revealed that the highest mortality (100%) was recorded in P0. This study revealed that inclusion of P. guajava extract in the diet of O. niloticus improved growth, nutrient utilization, immune system and survival of O. niloticus fingerlings.     

In vitro protein digestibility of different grow‐out stages of spotted rose snapper (Lutjanus guttatus,Steindachner, 1869)

The aim of this study was to compare in vitro protein digestibility between two groups of fish, at early (21 g) and late stages (400 g) of spotted rose snapper Lutjanus guttatus, to evaluate the degree of hydrolysis (DH) and total amino acid release (TAAR) using crude extracts from stomach, pyloric caeca and intestine of 13 protein ingredients including marine, animal and plant meals. Degree of hydrolysis and TAAR were measured by a pH‐Stat method, and the PAGE‐Zymogram was also used as complementary technique. Differences in DH were found between both grow‐out stages mainly in the alkaline hydrolysis phase. Fish and squid meals (marine sources) had the highest DH and TAAR, followed by porcine meat and poultry meal by‐products from recycling sources, and soybean and canola meals (plant sources), which represent better protein sources for use in practical diets. Stomach zymograms showed two pepsin isoforms in both grow‐out stages. Pyloric caeca and intestine zymograms showed five bands with proteolytic activity in the early grow‐out stage, whereas four additional bands were found in late grow‐out stage. Alkaline proteases were identified as serine and metalloproteases. Thus, L. guttatus presents an ontogenetically differentiated digestive enzyme pattern that modifies the DH and TAAR of different protein sources.     

Effects of replacing dietary fish oil and squid liver oil with vegetable oils on the growth,tissue fatty acid profile and total carotenoids of the giant freshwater prawn, Macrobrachium rosenbergii

A feeding trial was conducted to investigate the complete substitution of either fish oil (FO) or squid liver oil (SLO) with crude palm oil (CPO), canola oil (CO) sunflower oil (SFO) or linseed oil (LO), as the sole added lipid source in diets fed to triplicate groups of giant freshwater prawn, Macrobrachium rosenbergii (initial weight = 0.42 ± 0.01 g) for 6 weeks. Prawns fed the CO or SLO diets showed significantly higher (P < 0.05) specific growth rate than those fed the FO or CPO diets. The feed conversion ratio of the prawns was significantly better when fed the CO diet, compared with the FO, CPO, SFO and LO diets. The muscle eicosapentaenoic acid content of prawns fed the vegetable oil (VO) diets were not significantly different (P > 0.05) from those fed the FO diet, although all VO‐based diets led to a significantly lower docosahexaenoic acid content compared with prawns fed the FO or SLO diet. The whole‐body total carotenoid content was significantly lower for prawns fed the SLO diet compared with prawns on the CO or CPO diets. The successful use of VO instead of marine‐based oils in prawn diets will likely reduce feeding costs associated with M. rosenbergii aquaculture.