Effects of dietary supplementation of A3α‐peptidoglycan on the growth,immune response and defence of sea cucumber Apostichopus japonicus

To determine the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the growth, immune response and disease resistance of sea cucumber Apostichopus japonicus, a 70‐day feeding trial was conducted in this study. A total of 216 sea cucumbers were fed with four practical diets prepared from a commercial feed with different contents (0, 1.5, 2.5 and 4.0 g kg^?1) of A3α‐PG. The specific growth rate (SGR), total coelomocyte count (TCC), phagocytotic activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extracellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), peroxidase (POD) and superoxide dismutase (SOD), were measured at the end of the feeding trial. Finally, the animals were administered a 16‐day Vibrio splendidus challenge via intraperitoneal injection to test the potency of A3α‐PG on disease resistance. Compared with the control (0 g kg^?1 A3α‐PG), a significant increase (P < 0.05) in SGR was observed in the groups fed with 1.5 and 2.5 g kg^?1 A3a‐PG. The TCC, ranging from 7.25 × 10⁶ to 1.05 × 10⁷ cells mL^?1, was not significantly affected (P > 0.05) by A3α‐PG,. Coelomocyte phagocytotic activities in all of the A3α‐PG‐supplemented groups were significantly activated (P < 0.05), but no significant difference (P > 0.05) was observed. Sea cucumbers fed with 1.5 and 2.5 g kg^?1 A3α‐PG exhibited significant activation (P < 0.05) of EC/IC‐ACP, EC/IC‐ALP, and EC/IC‐POD activities. A significant increase in EC‐SOD activities (P < 0.05) was exhibited by all groups with A3α‐PG supplementation. The challenge test showed that animals fed with diets containing 2.5 and 4.0 g kg^?1 A3α‐PG had significantly lower cumulative mortalities compared with the control 16 days after exposure. All of the results presented here show that A3α‐PG can positively enhance the growth, immune response and disease resistance of sea cucumber, suggesting that dietary supplementation of A3α‐PG has potential applications in the health management of economic species of sea cucumber.     

Effects of continuous and alternate administration of β‐glucan and mannan‐oligosaccharide on the growth,immunity and resistance against Vibrio splendidus of sea cucumber Apostichopus japonicus (Selenka)

A 4‐week growth trial was conducted to compare the effects of different feeding strategies of dietary immunostimulants on the growth, immunity and resistance against Vibrio splendidus of sea cucumbers Apostichopus japonicus (Selenka). Six feeding strategies were set, including feeding immunostimulants‐free diet continuously (control), feeding dietary β‐glucan (1.25 g kg^?1 diet) continuously, feeding dietary mannan‐oligosaccharides (MOS; 2.00 g kg^?1 diet) continuously, feeding β‐glucan 2 days followed by MOS 5 days alternately, feeding β‐glucan 5 days followed by MOS 2 days alternately and feeding β‐glucan 7 days followed by MOS 7 days alternately. The sea cucumbers fed immunostimulants showed higher specific growth rate (SGR) and lower cumulative mortality than control (P < 0.05). When sea cucumbers were fed with β‐glucan continuously, total coelomocytes counts and superoxide anion were significantly higher than control on the 4th day (P < 0.05). However, these two immune parameters were not significantly higher than those in control after the 18th day (P > 0.05). While sea cucumbers continuously fed MOS, these two immune parameters were not significantly higher than control until the 15th day. All immune parameters of the sea cucumbers fed with β‐glucan and MOS alternately were significantly higher than those in control during the experiment (P < 0.05). The sea cucumbers fed with β‐glucan 7 days followed MOS 7 days alternately showed the highest SGR and second lowest cumulative mortality. It was suggested that this feeding strategy is most suitable for sea cucumbers.     

Dietary selenium requirement for on‐growing gibel carp (Carassius auratus gibelio var. CAS III)

A 12‐week growth trial was conducted in a flow‐through system to determine dietary selenium (Se) requirement for on‐growing gibel carp (initial body weight: 76.2 ± 0.05 g, mean ± SEM). Selenomethionine was supplemented to the basal diet to formulate seven semi‐purified diets containing 0.26, 0.58, 0.72, 1.14, 1.34, 1.73 and 2.09 mg Se kg^?1 diet. The results showed that plasma superoxide dismutase (SOD) activity significantly increased when fish were fed with 0.58 mg Se kg^?1 diet (P < 0.05) and then decreased at 2.09 mg Se kg^?1 diet (P < 0.05). Plasma T‐AOC activity was higher in fish fed with 0.72 mg Se kg^?1 diet (P < 0.05) and plasma malondialdehyde (MDA) was higher in fish fed with 0.26 mg Se kg^?1 diet (P < 0.05). When fish were fed 1.14 mg Se kg^?1 diet, hepatic GSH‐Px, T‐AOC, GSH and CAT activities were significantly higher than those fed with 0.26 mg Se kg^?1 diet (P < 0.05). Hepatic superoxide dismutase (SOD) activity was higher at 1.34 mg Se kg^?1 diet (P < 0.05). Fish liver Se concentrations were significantly higher when fed with 0.72 mg Se kg^?1 diet (P < 0.05) and then kept constant when Se ≥ 0.72 mg kg^?1 (P > 0.05). Whole‐body and muscle Se concentrations were higher when fed with 1.34 mg Se kg^?1 diet (P < 0.05) and kept a plateau when Se ≥ 1.34 mg kg^?1 (P > 0.05). In conclusion, based on broken‐line regression of hepatic Se concentrations, hepatic SOD activity and hepatic T‐AOC activity, dietary Se requirements for on‐growing gibel carp was 0.73 mg kg^?1, 1.12 mg kg^?1 and 1.19 mg kg^?1 diet respectively.     

A comparison between marine and terrestrial invertebrate meals for mirror carp (Cyprinus carpio) diets: Impact on growth,haematology and health

Invertebrate meals (e.g. polychaetes and insects) present novel and sustainable high‐quality nutrient sources for use in fish feed formulations. To test this innovative source, an eleven‐week feeding trial was conducted evaluating the effects of replacing the fishmeal (FM) component as an example of a superior protein source (FM CTRL) with ragworm meal (RW, Nereis virens) and/or silkworm pupae (SWP, Bombyx mori) in mirror carp (Cyprinus carpio) diets. Three experimental diets with partial replacement of FM (diets: RW + FM, SWP + FM and RW + SWP + FM) were formulated. All diets were formulated to be iso‐nitrogenous, iso‐lipidic and iso‐energetic. Growth performance and feed utilization indices were assessed, and the feeding trial concluded with the analysis of haematological parameters to provide an indication of carp physiological and health status. Mean weight gain was greatest in mirror carp fed RW + FM (60.83 fish^?1 day^?1; P < 0.05 vs. all other diets) followed by SWP + FM (40.62 g fish^?1 day^?1; P < 0.05 vs. all other diets). The least weight gain was achieved in fish fed FM + SWP + RW+ and FM CTRL (34.34 and 33.96 g fish^?1 day^?1, respectively; not significantly different from each other). Fish fed on RW + FM diet had significantly lower plasma ammonia concentrations than any other dietary groups (P = 0.04). Mirror carp fed on SWP + FM diet (111.52 units mL^?1) were observed to have a marked enhancement in alternative complement activity than FM CTRL (79.21 units mL^?1, P = 0.041). Both ragworm and silkworm pupae meal present attractive sustainable functional feed component in carp diets, with benefits on enhancing growth performance and specific physiological parameters.     

Influence of dietary phosphorus levels on growth,body composition,metabolic response and antioxidant capacity of juvenile snakehead (Channa argus × Channa maculata)

A study was conducted to estimate the optimum requirement of dietary phosphorus (P) for Channa argus × Channa maculata. Effects of dietary P levels on the tissue composition, serum biochemical parameters and antioxidant status were also examined. Five practical diets were formulated to contain graded levels (4.8 g kg^?1, 6.4 g kg^?1, 7.9 g kg^?1, 9.4 g kg^?1 and 11.0 g kg^?1) of available P from dietary ingredients and monocalcium phosphate (MCP). Each diet was randomly assigned to triplicate groups of 30 juvenile fish (initial body weight, 20.50 ± 0.53 g) for 8 weeks. The results showed that the specific growth rate (SGR) and weight gain (WG) were all significantly improved by dietary P up to 9.4 g kg^?1 (P < 0.05) and then levelled off beyond this level. Broken‐line analysis showed maximum weight gain (WG) was obtained at dietary available P concentrations of 9.6 g kg^?1. With the increase in dietary P level, protein efficiency rate (PER) increased significantly and reached a plateau, while the feed conversion ratio (FCR), the mesenteric lipid somatic index (MSI) and the whole‐body lipid content significantly reduced (P < 0.05). Dietary P levels also affected the mineralization (ash and P) of whole body, vertebrae and scale (P < 0.05). Quadratic analysis based on P contents in whole body, vertebrae, scale and ash content in vertebra indicated that the available P requirements were 10.4, 9.8, 10.0 and 10.3 g kg^?1, respectively. However, no differences were found in the whole‐body moisture, crude protein, serum calcium (Ca) contents or Ca/P value, as well as the viscerosomatic index (VSI) and hepatosomatic index (HSI) among all the treatments (P > 0.05). Triglyceride (TG), total cholesterol (TC), high‐density lipoprotein cholesterol (HDL‐C) and low‐density lipoprotein cholesterol (LDL‐C) decreased significantly, while serum P content, HDL‐C/TC and HDL‐C/LDL‐C value increased significantly with dietary available P levels (P < 0.05). No significant changes in superoxide dismutase activity and malondialdehyde (MDA) content were observed (P > 0.05), but serum catalase (CAT) and glutathione peroxidase (GPx) activities and the ratio of CAT/SOD and GPx/SOD increased significantly with increasing dietary P levels (P < 0.05). In conclusion, the optimal P requirement of juvenile snakehead in practical feed was 9.6 g kg^?1. Signs of P deficiency were characterized by poor growth, slightly reduced mineralization and the antioxidant capacity and an increase in body lipid content.     

Combination effects of dietary EPA and DHA plus alpha‐tocopherol: effects on performance and physiological status of Caspian brown trout (Salmo trutta caspius) fry

The effects of dietary n‐3 highly unsaturated fatty acids [eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA)] with α‐tocopherol on growth, non‐specific immune response and oxidative status were investigated in Caspian brown trout, Salmo trutta caspius, fry. Six experimental diets containing three different dietary levels of n‐3 HUFAs (low: 1 + 0.5% of total fatty acids, DHA+ EPA, medium: 2 + 1%, DHA + EPA, high: 4 + 2%, DHA + EPA) with two different levels of α‐tocopherol (low: 300 and high: 1000 mg kg^?1 diet) were prepared and named: LL, LH, ML, MH, HL and HH (HUFA/α‐tocopherol) groups, respectively. Diets were fed to triplicate groups of 60 fry with an initial weight of 600 ± 25 mg for 10 weeks. Results showed that increase in dietary DHA and EPA up to high level improved fry growth in terms of the body weight and specific growth rate, particularly when dietary α‐tocopherol levels were high, suggesting a higher antioxidant protection value when these fatty acids are high. At all dietary n‐3 HUFA levels, increase in α‐tocopherol from low to high level enhanced the alternative complement (ACH50) activity. Fry fed diets medium and high n‐3 HUFA displaying significantly higher lysozyme activity (P < 0.05). Moreover, fish fed medium or high levels of n‐3 HUFA had significantly lower prostaglandin E2 (PGE2) than those fed low n‐3 HUFA (P < 0.05). Significant differences in antioxidant enzymes (catalase, glutathione peroxidase, glutathione S‐transferase, glutathione reductase and superoxide dismutase) activity were also observed between groups, with higher activity in high levels of n‐3 HUFA (P < 0.05). Results of this study suggest that the effect of dietary n‐3 HUFA on examined non‐specific immunity parameters are not uniform; however, these impacts are closely related to the α‐tocopherol supplement and their interaction. In conclusion, increased dietary levels of n‐3 HUFA and α‐tocopherol would enhance growth performance and welfare of this species.     

Comparative study on dietary protein requirements for juvenile and pre‐adult gibel carp (Carassius auratus gibelio var. CAS III)

Two 8‐week growth trials were conducted in indoor recirculation system to evaluate the protein requirements for juvenile (3.70 ± 0.20 g) and pre‐adult (85.2 ± 0.70 g) gibel carp, Carassius auratus gibelio var. CAS III. Six isoenergetic diets were formulated for each trial using fish meal and casein as protein sources, and protein level was 250–450 g kg^?1 in Trial 1 and 200–450 g kg^?1 in Trial 2. With the increasing dietary protein, feeding rate (FR) and feed conversion ratio (FCR) significantly decreased (P < 0.05). Weight gain (WG) increased first and then reached a plateau in 330–450 g kg^?1 in Trial 1 (P > 0.05), while decreased after the maximum value in 350 g kg^?1 in Trial 2 (P < 0.05). Productive protein values (PPVs) were lower in 370–450 g kg^?1 in Trial 1 and 400–450 g kg^?1 in Trial 2 (P < 0.05). Increasing dietary protein level increased protein content and decreased lipid content in whole fish body and white muscle (P < 0.05). Apparent digestibility coefficient of dry matters (ADC_d) decreased, while apparent digestibility coefficient of protein (ADC_p) increased in 370–450 g kg^?1 in Trial 1 and 250–450 g kg^?1 in Trial 2 (P < 0.05). Trypsin activity significantly increased in 370–450 g kg^?1 in Trial 1 (P < 0.05) and was not affected in Trial 2 (P > 0.05). Hepatic alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in both trials increased when dietary protein was above 400 g kg^?1 (P < 0.05). Based on quadratic regression of WG, it was estimated that dietary protein requirement for maximum growth was 414 g kg^?1 (digestible protein of 376 g kg^?1) and 365 g kg^?1 (digestible protein of 324 g kg^?1) for juvenile (3.70 g) and pre‐adult gibel carp (85.2 g).     

Effects of dietary immunostimulant combination on the growth performance,non‐specific immunity and disease resistance of cobia,Rachycentron canadum (Linnaeus)

The aim of this study was to examine the effects of the immunostimulant combination (IC) containing β‐glucan, A3α‐peptidoglycan, vitamin C and vitamin E on the growth performance, non‐specific immunity and protection against Vibrio harveyi infection in cobia (Rachycentron canadum). Fish were fed diets containing six graded levels of IC (0, 1, 2, 3, 4 and 5 g kg^?1 diet) for 8 weeks. The results showed that the survival rate ranged from 81.1 to 84.4% with no significant difference among all the groups (P > 0.05) after the feeding experiment. Dietary IC significantly increased the specific growth rate (SGR), serum lysozyme, alternative complement pathway (ACH50) activity, phagocytosis percentage (PP) and respiratory burst activity of head kidney macrophages of cobia. Moreover, feeding of supplemented diets containing 3.0 g kg^?1 IC resulted in significantly lower mortality against the pathogens, V. harveyi compared with the control group. To elevate the growth and immune resistance ability of cobia, the optimal dose of dietary IC administration, determined by second‐order polynomial regression analysis was 3.43 and 2.71 g kg^?1 diet, respectively, on the basis of the SGR and mortality after challenge with V. harveyi.     

The effects of different proportions of the 17β‐estradiol and 17α‐methyltestosterone on growth,sex reversal and skin colouration of the electric blue hap (Sciaenochromis ahli Trewavas, 1935)

This study was performed to investigate the effects of 17β‐estradiol (ES) and 17α‐methyltestosterone (MT) on growth, development, survival, sex ratio and colour change in the electric blue hap (Sciaenochromis ahli Trewavas, 1935). The hormones were not supplemented to the control feed, while six other feeds were prepared by adding 20, 40 and 60 mg kg^?1 17β‐ES or 20, 40 and 60 mg kg^?1 17α‐MT to each, resulting in seven different feed treatments. Average live weight of the fish supplemented with these diets was 0.42 ± 0.04 g. At the end of the study, the highest weight gain was observed in fish fed 60 mg kg^?1 17α‐MT group (2.62 ± 0.11 g) and the difference with the groups fed with 17β‐ES was found to be significant. All fish fed 17α‐MT were male, while the rates of feminization in fish fed 17β‐ES at 20, 40, 60 mg kg^?1 were 91.11%, 88.88% and 93.33% respectively. Survival rates were respectively determined as 80%, 95.56%, 84.44%, 93.33%, 77.78%, 84.44% and 84.44% for the control, 20, 40, 60 mg kg^?1 17β‐ES and 20, 40, 60 mg kg^?1 17α‐MT treatments. The best colouration was achieved in the 17α‐MT groups (P < 0.05). The L* values varied between 32.98 ± 4.44 and 61.35 ± 2.19, a* values between ?7.06 ± 0.22 and ?3.42 ± 0.11, and b* values between ?7.74 ± 0.10 and 11.65 ± 0.03, while Chroma (C*) and Hue (H°_ab) angle values varied between 7.54 ± 0.22 and 13.60 ± 0.01 and between 119.76 ± 0.05 and 239.73 ± 4.86. In conclusion, the 17α‐MT feeding was found to have a greater effect on the growth, feed conversion ratio, masculunization and pigmentation of the electric blue haps than the 17β‐ES treatment.     

Effects of dietary live or heat‐inactivated autochthonous Bacillus pumilus SE5 on growth performance,immune responses and immune gene expression in grouper Epinephelus coioides

To evaluate the possible dietary application of live and heat‐inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 10⁸ CFU g^?1 live (T1) and heat‐inactivated B. pumilus SE5 (T2). The heat‐inactivated probiotic significantly improved the final weight, weight gain (WG) and specific growth rate (SGR) at day 60 and significantly decreased the feed conversion ratio (FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 (P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat‐inactivated probiotic for 60 days (P < 0.05). Furthermore, the heat‐inactivated probiotic remarkably up‐regulated expression of TLR2 and pro‐inflammatory cytokines (IL‐8 and IL‐1β) in head kidney (P < 0.05), but the viable probiotic failed to do so. These results indicated that heat‐inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides.     

Effect of graded levels of dietary thiamine on the growth performance,body composition and haemato‐biochemical parameters of juvenile Sclizothorax prenanti

This study was conducted to determine dietary thiamine requirement of juvenile Sclizothorax prenanti and evaluate the effect of dietary thiamine levels on growth performance, body composition and haemato‐biochemical parameters for this fish species. The seven experimental diets were formulated to contain the graded levels of thiamine (0, 10, 20, 30, 40, 60 and 100 mg kg^?1 diet, respectively), providing the actual dietary thiamine values of 0.31 (control), 9.82, 21.49, 29.83, 41.66, 62.24 and 114.58 mg kg^?1 diet, respectively. Each diet was assigned to three replicate groups of S. prenanti (initial body weight: 13.46 ± 0.28 g, means ± SD) for 60 days. Increasing dietary thiamine level up to 21.49 mg kg^?1 diet increased weight gain rate (WGR), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) (P < 0.05), beyond which they remained nearly unchanged. Similarly, hepatic thiamine concentration and several serum biochemical parameters (transketolase activity, triglyceride and total cholesterol contents) increased with increasing levels of thiamine up to 21.49 mg kg^?1 diet (P < 0.05) and, thereafter, remained almost constant. However, no significant differences in body composition (moisture, protein, lipid and ash contents) were found among dietary thiamine treatments (P > 0.05). Analysis by the broken‐line regression of WGR, SGR, FE, PER, hepatic thiamine concentration and serum transketolase activity indicated that dietary thiamine requirements in juvenile S. prenanti were 18.45–25.91 mg kg^?1 diet.     

Depletion of florfenicol amine in tilapia (Oreochromis sp.) maintained in a recirculating aquaculture system following Aquaflor®‐medicated feed therapy

Aquaflor^® [50% w w^?1 florfenicol (FFC)], is approved for use in freshwater‐reared warmwater finfish which include tilapia Oreochromis spp. in the United States to control mortality from Streptococcus iniae. The depletion of florfenicol amine (FFA), the marker residue of FFC, was evaluated after feeding FFC‐medicated feed to deliver a nominal 20 mg FFC kg^?1 BW d^?1 dose (1.33× the label use of 15 mg FFC kg^?1 BW d^?1) to Nile tilapia O. niloticus and hybrid tilapia O. niloticus × O. aureus held in a recirculating aquaculture system (RAS) at production‐scale holding densities. Florfenicol amine concentrations were determined in fillets taken from 10 fish before dosing and from 20 fish at nine time points after dosing (from 1 to 240 h post‐dosing). Water samples were assayed for FFC before, during and after the dosing period. Parameters monitored included daily feed consumption and biofilter function (levels of ammonia, nitrite and nitrate). Mean fillet FFA concentration decreased from 13.77 μg g^?1 at 1‐h post dosing to 0.39 μg g^?1 at 240‐h post dosing. Water FFC concentration decreased from a maximum of 1400 ng mL^?1 at 1 day post‐dosing to 847 ng mL^?1 at 240 h post‐dosing. There were no adverse effects noted on fish, feed consumption or biofilter function associated with FFC‐medicated feed administration to tilapia.     

Supplementation of sodium chloride in diets to improve the meat quality of Pacific white shrimp,Litopenaeus vannamei,reared in low‐salinity water

To investigate the effect of dietary sodium chloride (NaCl) on meat quality of white shrimp (Litopenaeus vannamei) reared in low‐salinity (2 g L^?1) water, shrimp were distributed into four groups (treatments T‐1, T‐2, T‐3 and control) with three replicates. All shrimps were completely randomised stocked into 12 tanks at an initial density of 40 shrimps per tank. Diets for the control, T‐1, T‐2 and T‐3 groups consisted of the basal diet supplemented with 0 g kg^?1, 10 g kg^?1, 20 g kg^?1 and 40 g kg^?1 of NaCl respectively. After 50 days, shrimps in T‐3 showed significantly better (P < 0.05) moisture, crude protein and ash than those of the control and T‐1. Higher muscle Na content was observed (P < 0.05) in T‐3 than that of the control. Significant increases (P < 0.05) in contents of inosinic acid, total free amino acid (TFAA) and essential free amino acid (EFAA) were also found in T‐3. Texture assays showed significant differences (P < 0.05) in hardness, adhesiveness and springiness between group T‐3 as compared with those of T‐1 and control. It indicated that dietary supplementation of NaCl appeared to be a promising practice to improve meat quality of white shrimp reared in low‐salinity waters.     

Inhibition of Vibrio spp. by 2‐Hydroxyethyl‐11‐hydroxyhexadec‐9‐enoate of Marine Cyanobacterium Leptolyngbya sp. LT19

Seven marine cyanobacteria were isolated from two regions of the Gulf of Thailand and evaluated by the agar diffusion method for antibacterial activity. Inhibitory compound was purified from the crude methanol extract and its structure was elucidated based on extensive spectroscopic analysis, including IR, 1D and 2D NMR spectra as well as mass spectrometry. A novel antimicrobial compound produced by the marine cyanobacterium Leptolyngbya sp. LT19 was identified to be a 2‐hydroxyethyl‐11‐hydroxyhexadec‐9‐enoate which has so far never been reported in microorganisms. Biological assays revealed that this novel compound exhibited antibacterial activities against the Gram‐negative, persistent shrimp pathogens, Vibrio harveyi and V. parahaemolyticus with minimal inhibitory concentration of 250–1000 and 350–1000 μg mL^?1 respectively.     

Growth,digestive and absorptive abilities and antioxidative capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idellus Val.) fed graded levels of dietary manganese

To study the effects of manganese on growth performance, digestive and absorptive abilities, as well as the antioxidative capacity in the hepatopancreas and intestine, young grass carp (Ctenopharyngodon idellus Val.) (264 ± 1 g) were fed diets containing graded levels of manganese at 3.65 (control), 8.62, 13.48, 18.24, 22.97 and 27.86 mg kg^?1 diet for 8 weeks. Per cent weight gain (PWG) and feed intake were the poorest in fish fed the basal diet (P < 0.05). The activities of trypsin, lipase and alkaline phosphatase in the intestine were significantly enhanced with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05). Additionally, in the hepatopancreas and intestine, the protein carbonyl and malondialdehyde contents were the lowest in fish fed the diet with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05), while the anti‐hydroxyl radical capacities, manganese superoxide dismutase (MnSOD), glutathione peroxidase and glutathione‐S‐transferase activities were significantly enhanced with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05). Moreover, the catalase activity and glutathione content in the intestine were the highest in fish fed the diet with dietary manganese level at 18.24 mg kg^?1 diet (P < 0.05). These results indicated that optimum dietary manganese promoted growth, enhanced the digestive and absorptive abilities, and improved the antioxidative capacity in young grass carp. Based on the quadratic regression analysis for PWG and intestinal MnSOD activity, the manganese requirements for young grass carp with the initial body weight of 264 g were 16.91 and 18.21 mg kg^?1 diet respectively.     

Dietary methionine requirement of juvenile Pseudobagrus ussuriensis

An 8‐week feeding trial was conducted to determine the optimum dietary methionine (Met) requirement of juvenile Pseudobagrus ussuriensis with an initial average weight of 0.60 g reared in indoor flow‐through and aerated aquaria. Six isonitrogenous (430 g kg^?1 protein) and isolipidic (50 g kg^?1 lipid) test diets were formulated to contain graded levels of crystalline L‐methionine (4.9, 9.0, 11.8, 14.2, 18.1 and 20.8 g kg^?1 dry diets, respectively) at a constant dietary cystine level of 2.5 g kg^?1 dry diets. Equal amino acid nitrogen was maintained by replacing methionine with non‐essential amino acid mixture. Fish were randomly allotted to 18 aquaria (1.0 × 0.5 × 0.8 m) with 50 fish to each glass aquarium. Fish were fed twice daily (08:00 and 16:00) to apparent satiation. No significant difference was observed in survival of fish (84.67–91.33%). Specific growth rate (SGR), weight gain (WG), feed conversion ratio (FCR), protein productive value (PPV) and protein efficiency ratio (PER) were significantly affected by different dietary methionine levels (P < 0.05). WG, SGR PPV and PER increased, while FCR decreased with increasing dietary methionine level from 4.9 to 11.8 g kg^?1 (P < 0.05). However, with further increase from 11.8 to 20.8 g kg^?1, WG, SGR PPV and PER significantly decreased, FCR increased (P < 0.05). The whole body and muscle composition were affected by different dietary methionine levels (P < 0.05). Condition factor (CF) increased with increasing dietary methionine levels up to 11.8 g kg^?1 (P < 0.05) and after 11.8 g kg^?1 methionine diet, but not significant, declines were observed (P > 0.05). Hepatosomatic index (HSI) of the 4.9, 9.0, 11.8 and 14.2 g kg^?1 Met diets was significantly higher than that of fish fed diets 18.1 and 20.8 g kg^?1 Met diets (P < 0.05). Viscerosomatic index (VSI) of the 4.9, 9.0 and 11.8 g kg^?1 Met diets was significantly higher than that of fish fed diets 14.2, 18.1 and 20.8 g kg^?1 Met diets (P < 0.05). Quadratic regression analysis of WG and PER against dietary methionine levels indicated that the optimal dietary methionine requirement for maximum growth and feed utilization of juvenile Pseudobagrus ussuriensis was 14.3 and 14.1 g kg^?1 dry diet (35.3 and 34.8 g kg^?1 dietary protein), respectively, in the presence of 2.5 g kg^?1 dry diets cystine.     

Dietary biofloc supplementation in black tiger shrimp,Penaeus monodon: effects on immunity,antioxidant and metabolic enzyme activities

A 60‐day indoor experiment was conducted to study the effect of dietary supplementation of biofloc on metabolic enzyme activities and immune responses in Penaeus monodon juveniles. Biofloc developed in indoor fibreglass‐reinforced plastic (FRP) tanks (1000 L) was used as dietary supplement in P. monodon (2.90 ± 0.10 g) reared in 1000‐L FRP tanks. Graded level of dried biofloc was included in shrimp basal diets, 0% (control, B0), 4% (B4), 8% (B8) and 12% (B12). The level of metabolic enzymes like malate dehydrogenase (MDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) was not significantly different with control up to 8% dietary supplementation. A higher level of total haemocyte count (THC) was noticed in B8 (22.16 ± 2.17 × 10⁶ cells mL^?1) and B4 (21.11 ± 0.56 × 10⁶ cells mL^?1) compared with control, C (14.61 ± 2.74 × 10⁶ cells mL^?1). Biofloc‐supplemented groups recorded significantly higher (P < 0.05) serum SOD and catalase activity (P < 0.01) in comparison with control. The groups fed with 4% dietary biofloc supplement recorded highest relative percentage survival (RPS), 45% after challenge with Vibrio harveyi followed by 36% and 27% RPS in B8 and B12 groups. Based on these results, it can be concluded that supplementation of biofloc even at 4% level in the feed improves immune responses and metabolic activities in black tiger shrimp juveniles.     

Synchronization of ovulation in cultured northern whitefish (Coregonus peled,Gmelin 1788) using [D‐Arg6Pro9Net]‐sGnRH analogue and its effect on egg quality

Female northern whitefish were treated with salmon [D‐Arg⁶Pro⁹Net]‐sGnRHa emulsified in Freund′s incomplete adjuvant as a sustained release treatment (sGnRHa‐FIA) or with [D‐Arg⁶Pro⁹Net]‐sGnRHa dissolved in physiological saline as acute release treatment (sGnRHa‐PS). Females in four experimental groups (A, B, C, D) and one control group (E) were injected intraperitoneally as follows: A and B – sGnRHa‐FIA at doses of 50 and 25 μg kg^?1, respectively; C – double injection (DI) of sGnRHa‐PS at 25 μg kg^?1 administered 3 days apart; D – single injection (SI) of sGnRHa‐PS at 25 μg kg^?1; and E – control group receiving a 1:1 mixture of FIA and physiological saline. All treatments induced and synchronized ovulation. Mean time to ovulation was significantly reduced (P < 0.05) in GnRHa‐treated groups compared with control. Hormone treatments did not affect the relative fecundity. Slight differences were found in ovarian fluid pH between group A and D (P < 0.05). Except for group C, egg size was significantly reduced (P < 0.01) in hormonally synchronized groups compared with the control. Survival to the eyed stage of eggs from females in groups A, B and C was significantly lower (P < 0.01) than in groups D and E. Per cent hatched alevins was lower (P < 0.01) in groups A, B and C than in groups D and E. We conclude that synchronization of ovulation in northern whitefish can be induced by a single acute injection of [D‐Arg⁶Pro⁹NEt]‐sGnRHa at 25 μg kg^?1 BW if given near the natural spawning time determined by water temperature below 2°C.     

Effects of aqueous Na/K and dietary K on growth and physiological characters of the Pacific white shrimp,Litopenaeus vannamei,reared in low‐salt well water

The synchronous effects of aqueous Na/K and dietary potassium (K⁺) on growth and physiological characters was studied on the Pacific white shrimp (Litopenaeus vannamei) reared in low‐salinity well water (4 ppt) for 8 weeks with initial weight of 0.28 ± 0.01 g. Three practical diets were formulated with supplement of 0, 0.3%, 0.6% K⁺ which contained 1.29 g/100 g, 1.60 g/100 g, 1.93 g/100 g K⁺ respectively. The supplement of K⁺ to the low‐salinity well water was 10, 20, 40 mg L^?1 which formed Na:K ratios of 42:1, 33:1, 23:1 respectively. Results showed that when the aqueous Na:K ratio was 42 and dietary K⁺ was 1.93 g/100 g K⁺, the WGR and PER of L. vannamei were the highest and the FCR was lower than that of others (P < 0.01). Supplement of K⁺ into well water and diets did not showed significant effects on haemolymph ammonia‐N, uric acid, urea content (P > 0.05), but had a extremely significant effect on arginase activity and Cl^? concentration (P < 0.01). Moreover, similar results were observed in alkaline phosphatase (ALP), bacteriolytic activity (LSZ) and respiratory burst activity (O₂^?) (P < 0.05). These results suggested that aqueous Na/K in the low‐salt well water and dietary K had significant synergistic effect on the growth, osmoregulation and immunity of L. vannamei. Concluded from the growth performance, nitrogen metabolism, osmoregulation and immunity, as the Na/K in the low‐salinity well water descended from 42 to 23, the requirement of dietary K⁺ was also decreased.     

Effectiveness of l‐ascorbyl‐2‐polyphosphate as an ascorbic acid source for sea cucumber,Apostichopus japonicus

A feeding trial was conducted to determine the adequate dietary ascorbic acid (AsA) levels and the effects on growth, meat quality and antioxidant status of sea cucumber (10.04 ± 0.06 g), Apostichopus japonicus. l ‐ascorbyl‐2‐polyphosphate (35% AsA equivalent) was supplemented separately to the basal diet to obtain five AsA levels, 0, 598, 1473, 4676 and 14340 mg kg^?1 diet respectively. After 60‐day feeding trial, the sea cucumbers fed diets containing 598 and 1473 mg AsA kg^?1 showed significantly higher (P < 0.05) body weight gain and specific growth rate values than the sea cucumbers fed control diets. The sea cucumbers fed diets containing 1473 and 4676 mg AsA kg^?1 showed significantly higher (P < 0.05) hydroxyproline contents than those of the sea cucumbers fed diets containing 0 and 598 mg AsA kg^?1. Antioxidant enzymes such as total antioxidant capacity, superoxide dismutase and glutathione peroxidase showed increasing trends with the increasing dietary AsA levels, but no significant differences (P > 0.05) were observed when the sea cucumbers fed diets with high dietary AsA levels. The content of malondialdehyde had the opposite trend of antioxidant enzymes. In conclusion, the adequate dietary AsA level focusing on growth performance of sea cucumber is between 598 and 1473 mg kg^?1 diet. Furthermore, high level of dietary AsA (between 598 and 4676 mg kg^?1 diet) improved meat quality and antioxidant status.