Improvement of reproductive indices,lysozyme activity,and disease resistance in live‐bearing ornamental fish,Poecilia latipinna using Artemia supplementation with treated yeast cell,Saccharomyces cerevisiae

In the present study, the effects of Artemia supplemented with 2‐β‐mercapto‐ethanol (β‐ME) treated yeast cell, Saccharomyces cerevisiae, on growth and reproductive performance, lysozyme activity, and disease resistance to Aeromonas hydrophila of freshwater ornamental species, Poecilia latipinna, were investigated. Within 60 days, molly fish were fed with three treatments including commercial food (T1), un‐supplemented Artemia (T2), and Artemia supplemented with β‐ME‐treated yeast cell (at concentration of 4 × 10⁷ CFU/L of water) (T3). After the feeding period, the fish were exposed to 100 μl of a suspension (1.1 × 10⁷ cells/ml) of A. hydrophila (BCCM5/LMG3279) and the cumulative mortality rates were recorded for 12 days. No significant difference was found between survival rate and growth performance of P. latipinna except for weight gain that was higher in fish fed through Artemia supplemented with β‐ME‐treated yeast cell compared to control group. Fecundity rate was significantly improved in fish fed using T3 with the maximum amount of 49.5 ± 2.29 per female (p < 0.05). Besides, lysozyme activity was significantly increased in group 3 (p < 0.05). Moreover, lowest fish mortality was significantly observed in this treatment (p < 0.05). In addition, the number of colonies formed by yeast cell in T3 (634 × 10³ CFU/g intestine) showed significant difference with other treatments (p < 0.05). In sum, Artemia enriched with β‐ME‐treated yeast improved reproductive indices, immune responses, and resistance against A. hydrophila of P. latipinna.     

Dietary supplementation effects of Pediococcus acidilactici as probiotic on growth performance,digestive enzyme activities and immunity response in zebrafish (Danio rerio)

This study was carried out to assess the effects of Pediococcus acidilactici on zebrafish (Danio rerio). Different levels of P. acidilactici including 0, 1 × 10⁶, 2 × 10⁶, 4 × 10⁶ and 8 × 10⁶ colony‐forming unit per g of the diet (cfug^?1) were examined in fish with 120 ± 10 mg weight for 60 days in a completely randomized design. The results showed that the best growth indices were recorded in group 4 × 10⁶ cfug^?1 (p < 0.05). The highest number of total viable count and lactic acid bacteria of intestine were found in group 4 × 10⁶ cfug^?1 (p < 0.05). The maximum activity of digestive enzymes including amylase, lipase, protease and alkaline phosphatase was observed in group 4 × 10⁶ cfug^?1. The highest activity for superoxide dismutase was recorded in group 4 × 10⁶ cfug^?1 while catalase, glutathione peroxidase and glutathione reductase showed the highest activity in group 8 × 10⁶ cfug^?1. The most growth inhibition zone of Aeromonas hydrophylla, Flavobacterium columnare, Vibrio anguillarum and Edwardsiella tarda was found in group 4 × 10⁶ cfug^‐1 (p < 0.05). Therefore, P. acidilactici as a probiotic improved growth and immunity of the zebrafish and could be used by zebrafish farmers.     

Growth and immune response of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae fed diets containing Chlorella vulgaris (Beijerinck)

A 50‐day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Iso‐nitrogenous and iso‐lipidic test diets were prepared using a fishmeal‐based‐positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30‐L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95‐L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.     

The obvious heterosis and genetic characters of intergeneric cross and backcross juveniles between blunt snout bream (Megalobrama amblycephala) and topmouth culter (Culter alburnus)

In this study, we used artificial insemination to generate hybrid groups of fish [MC‐F₁(MA♀×CA♂) and MC‐F₂(MC‐F₁♀×♂)] by intergeneric crosses of Megalobrama amblycephala (MA) and Culter alburnus (CA); sequential backcrosses [CAM‐B₁ (CA♀×MC‐F₁♂) and MCC‐B₁ (MC‐F₁♀×CA♂)] were also performed. All these hybrids showed high rates of fertilization, hatching and survival (p > 0.05). For genetic traits, compared with those of the M. amblycephala and C. alburnus parental lines (Table 1), the fertilization rate, hatching rate and 7‐day survival rate of MC‐F1(MA♀×CA♂), MC‐F2(MC‐F1♀×♂), CAM‐B1 (CA♀×MC‐F1♂) and MCC‐B1 (MC‐F1♀×CA♂) by artificial insemination exhibited similar high rates (p > 0.05). The morphology of the four hybrids MC‐F₁/F_2, CAM‐B₁ and MCC‐B₁ were intermediate between those of their parents. Compared with their parents of MA and CA, weight gain rate (WG), specific growth rate (SGR) and protein efficiency ratio (PER) of hybrids MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ were significantly (p < 0.05) increased and feed conversion ratio (FCR) was significantly (p < 0.05) decreased after 3 months feeding. Moreover, protein content of muscle for MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ was significantly (p < 0.05) higher and carbohydrate content of muscle was significantly (p < 0.05) lower than their parents. The females and males of the four hybrids had normal gonadal development. In this study, we successfully generated intergeneric and backcross hybridization lines with fertile potential among fish of the Cultrinae subfamily and these hybrids had obvious heterosis in terms of growth performance, feed utilization and muscle quality.     

Effect of probiotics on immune competence of giant freshwater prawn Macrobrachium rosenbergii

This study evaluates the effect of probiotics, Zymetin, on the immune efficiency of Macrobrachium rosenbergii juvenile against pathogenic Vibriospp. and Aeromonasspp. The experiment was conducted in glass aquaria with same level of feeding under different treatments, that is, negative control (Cn), positive control with Vibrio spp. (Cv) and Aeromonasspp. (Ca), prawn juveniles fed with probiotics (Zymetin) at 5 g/kg of feed but without pathogen (T1), probiotic fed prawn challenged with Vibrio spp. (T2) and Aeromonasspp. (T3). The results demonstrated that T1 showed higher survival rate, total haemocyte count, non‐granular haemocyte count, phagocytic activity, and clearance efficiency compared to other treatment groups. In contrast, decreased number of small and large granular haemocyte was observed in T1. Despite that, THC was found to be significantly different (p < 0.05) among all the controls and lowest was in Cv, followed by Ca. Besides, phagocytic activity and clearance efficiency against Vibrio spp. and Aeromonasspp. decreased significantly in Cn, Ca, and Cv fed M. rosenbergiijuvenile, while the values were found to be higher in T1 followed by T2 and T3, in that order. In addition, the challenge test showed an increasing trend of total and beneficial bacterial density as well as declining trend of some harmful bacteria in the water and gut of prawn in all the groups tested (p < 0.05).     

Dietary supplementation with Bacillus subtilis LT3‐1 enhance the growth,immunity and disease resistance against Streptococcus agalactiae infection in genetically improved farmed tilapia,Oreochromis niloticus

A feeding trial was conducted to investigate the effect of different levels of Bacillus subtilis LT3‐1 in diets on growth, immune parameters, intestinal morphology and disease resistance in genetically improved farmed tilapia, Oreochromis niloticus. Fish (46.91 ± 0.17 g) were fed with a basal diet supplemented with B. subtilis LT3‐1 at 0 (B0), 3.8 × 10¹⁰ (B1), 7.6 × 10¹⁰ (B2), 1.14 × 10¹¹ (B3) and 1.52 × 10¹¹ (B4) CFU kg^?1 for 6 weeks. The results showed that the weight gain of fish in B1 group was significantly enhanced compared to that in B0 group (p < 0.05). The addition of B. subtilis significantly affected serum biochemical indices (total protein, albumin, aspartate aminotransferase, alkaline phosphatase). Besides, the haematocrit, total counts of red and white blood cells, as well as the serum catalase and lysozyme activities, were increased, whereas the serum malondialdehyde, the serum immunoglobulin M and complement three contents were reduced. Parameters for intestinal morphology suggested a healthier intestine for the fish fed B. subtilis‐supplemented diets than fish fed the control diet. The survival rate after Streptococcus agalactiae challenge increased in tilapia fed with B. subtilis. The present study demonstrated B. subtilis can effectively improve growth, immunological status and resistance against S. agalactiae infection in tilapia farming.     

Histopathological changes in giant freshwater prawn Macrobrachium rosenbergii (de Man 1879) fed with probiotic Bacillus licheniformis upon challenge with Vibrio alginolyticus

The effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the histopathological changes in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g) challenged with known pathogenic strain of Vibrio alginolyticus are reported. Two isocaloric basal diets supplemented with probiotic bacteria B. licheniformis (1.0 × 10⁹ cfu/g feed) and other without probiotic supplementation were fed to the M. rosenbergii juveniles for 45 days. The histological observations revealed no significant changes in the hepatopancreas and gut tissues of both the experimental and the control groups which indicate that the present bacterium is a safe candidate probiont for the host. Prawns were challenged with V. alginolyticus after 45 days of feeding with probiotic diet. The histopathological studies of the hepatopancreas revealed that M. rosenbergii fed with probiotic‐supplemented diet showed less changes as compared to the prawns fed with control diet on second and fourth day of post‐experimental challenge with V. alginolyticus. Histopathological observations revealed that the gills of the prawns fed with control diet were severely affected in comparison to the prawns fed with probiotic‐supplemented diet after challenging with V. alginolyticus. Results from this study revealed the improved protection by dietary incorporation of B. licheniformis in reducing the histopathological manifestations due to V. alginolyticus infection in freshwater prawn.     

Effect of dietary soybean lecithin on growth parameters,digestive enzyme activity,antioxidative status and mucosal immune responses of common carp (Cyprinus carpio)

Common carp, Cyprinus carpio (13.54 ± 0.48 g), were fed soybean lecithin‐incorporated diets for 8 weeks. After the feeding trials, the different groups of fishes were assessed by survival, growth parameters, digestive enzymes, antioxidant enzyme activities, mucosal immune parameters, etc. The lecithin‐incorporated diets (10–30 g kg^–1) helped to increase the weight gain significantly (p < .05) compared with control groups; activity of digestive enzymes was also significantly (p < .05) higher compared with experimental groups. The antioxidant enzymes including catalase and serum superoxide dismutase were significantly (p < .05) higher in the C. carpio fed lecithin‐incorporated feed than the ones fed the control feed. The serum thiobarbituric acid‐reactive substances and serum glutathione peroxidase activity did not differ significantly (p > .05) between control and experimental groups. The positive correlation supported that antibacterial activity was significantly (p < .05) increased with increasing concentration of lecithin in the diets. Among the different mucosal immune parameters, lysozyme, protease and esterase activities were significantly (p < .05) increased with lecithin‐incorporated diets. The results of this study confirm that 20 and 30 g kg^–1 lecithin inclusion in diets helped to enhance the activities of digestive enzymes, antioxidant enzymes, antibacterial factors and mucosal immune parameters in C. carpio.     

Effect of dietary sodium butyrate on growth performance,enzyme activities and intestinal proliferation‐related gene expression of juvenile golden pompano Trachinotus ovatus

The present study was conducted to investigate the effects of dietary sodium butyrate on growth performance, intestine enzyme activities and intestinal proliferation‐related gene expression of juvenile golden pompano Trachinotus ovatus. A basal diet was supplemented with sodium butyrate at 0.0 (control), 0.25, 0.5, 1, 2 and 4 g/kg feed for 8 weeks. The final body weight, weight gain (WG), specific growth rate (SGR) and condition factor (CF) increased with increasing dietary sodium butyrate up to 2.0 g/kg, and thereafter declined, while feed conversion ratio exhibited an opposite trend. Compared with the control, the 2.0 g/kg sodium butyrate group had higher condition factor (CF) significantly (p < 0.05). Whole body ash decreased with increasing dietary sodium butyrate level, with the lowest whole body ash content in 4.0 g/kg sodium butyrate (p < 0.05). Compared with the control, the 2.0 g/kg sodium butyrate group had significantly increased plasma glucose, cholesterol, albumin level, A/G ratio, ALT and AST contents (p < 0.05), while significantly decreased plasma ALT/AST ratio (p < 0.05). As for the intestinal digestive and brush border enzymes activities, compared with the control, the 2.0 and 4.0 g/kg sodium butyrate groups had significantly increased intestinal protease, amylase, AKP and Na⁺‐K⁺‐ATPase activities (p < 0.05), respectively. The relative level of intestinal CDX2 mRNA of fish significantly increased with dietary sodium butyrate level. Compared with the control, the 2.0 g/kg sodium butyrate groups had significantly increased the expression of intestinal CDX2 and CREB mRNA (p < 0.05), respectively. In conclusion, these results suggested that the optimum sodium butyrate level for juvenile golden pompano could be 2.0 g/kg of the diet.     

Effects of dietary supplementation of probiotics on the growth,activities of digestive and non‐specific immune enzymes in hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀)

Effects of Bacillus cereus BC‐01, Lactobacillus acidophilus LAG01, Clostridium butyricum CBG01 and their combinations as supplementation on the growth performance, digestive enzyme activities and serum non‐specific immunity of hybrid grouper (Epinephelus lanceolatus ♂×E. fuscoguttatus ♀) were assessed. Seven different diets, that is one control diet (basal feed without any probiotics, CT) and six treatment diets containing single B. cereus (Bs), L. acidophilus (Ls) and C. butyricum (Cs) at 1.0 × 10⁹ cfu/kg feed, and also their combinations in equal proportion at 1.0, 2.0 and 3.0 × 10⁹ cfu/kg feed (BLC1, BLC2 and BLC3) were prepared respectively. After 60‐day feeding trial, the final weight, specific growth rate,food consumption, food conversion efficiency and apparent digestibility coefficient of fish in Ls and BLC3 were significantly higher compared with the control (p < .05). The activities of pepsin and trypsin in the intestine of fish for Ls and BLC3 were significantly higher relative to the control (p < .05). Relative to controls, significantly enhanced amylase and lipase activities in proximal intestine except for Cs and BLC1 and lipase activities in distal intestine except for Cs were observed (p < .05). Meanwhile, activities of superoxide dismutase in the serum of fish for all treatments, lysozyme and catalase in Ls and BLC3, and glutathione peroxidase except for Cs were significantly enhanced (p < .05). Based on the above, dietary supplementation of single L. acidophilus at 1.0 × 10⁹ cfu/kg or combination of three strains at 3.0 × 10⁹ cfu/kg for hybrid grouper is recommended.     

Effects of different dietary lipids on growth,body composition and lipid metabolism‐related enzymes and genes in juvenile largemouth bass,Micropterus salmoides

This study investigates the effects of different lipids on growth, body composition and lipid metabolism of largemouth sea bass fish Micropterus salmoides. A total of 360 juvenile M. salmoides (mean ± SD mass = 33.83 ± 0.15 g) were randomly stocked into 12 tanks of 0.5 m³ volume for 8 weeks. Four replicates were made in each group, which were fed one of three diets containing fish oil (FO), soybean oil (SO) or lard oil (LO). The weight gain rate and specific growth rate did not differ among the groups (p > 0.05). Fish oil fish had the lowest condition factor (p < 0.05) and highest serum glucose content (p < 0.05). Crude lipid contents in the whole body and in the liver and muscle of FO fish were significantly lower than in the SO and LO groups (p < 0.05). The fatty acid composition of whole‐body lipids was closely correlated with that of the diet. The carnitine palmitoyltransferase 1 (cpt1) activity in the FO group was significantly higher than those in the SO and LO groups (p < 0.05). No significant differences in fatty acid synthase (fasn) activity were observed among the groups (p > 0.05). The Cpt1 and fasn gene expression levels in the FO group were significantly higher than those of the SO and LO groups (p < 0.05). The apolipoprotein B100 gene expression level was significantly higher in the SO group than in the FO group (p < 0.05). Fatty acid‐binding protein 1 gene expression levels in the FO and SO groups were not different (p > 0.05) but were both higher than that of the LO group (p < 0.05). The delta‐6 fatty acyl desaturase gene expression level in the LO group was significantly higher than that in the FO group (p < 0.05), but lower than that in the SO group (p < 0.05). It can be concluded that FO can be completely replaced by SO or LO in the M. salmoides diet, at least within the 8‐week culture period. Different types of dietary lipids significantly affect body condition and hepatic lipid metabolism in M. salmoides.     

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.     

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

An investigation on the gamete quality of Black Sea trout (Salmo trutta labrax) broodstock fed with mealworm (Tenebrio molitor)

In this study, the effect of combined feeding Black Sea trout (Salmo trutta labrax) broodstock with commercial feed and mealworm larvae (Tenebrio molitor) for 50 days before the breeding period in order to evaluate the gamete quality. While the control group was fed with commercial feed, mealworm larvae were given as an additional protein source 2 days a week and 3 days a week to the experimental groups formed from female and male individuals. In addition to the growth parameters, the number of eggs (number/individual) and the egg diameter (mm), sperm volume (ml), density of spermatozoa (×10⁹ cell/ml), total motility (%), progressive motility (%) and average curvilinear velocity (VCL, µm/sec) values were determined. At the end of the study, the highest live weight gain was found as 46.2 g in the control group of male. The egg diameters were 4.3 ± 1.8 mm and 4.5 ± 1.4 mm in the worm treatment groups fed twice and three times with mealworm respectively. As a result, it was determined that feeding fish with mealworm larvae as an addition to the commercial diet in female individuals did not affect the amount of eggs (p > .05), but the egg diameters were significantly smaller in the control group than the mealworm groups (p < .05). In spermatological characteristics, only the amount of sperm was different between the groups (p < .05); all other parameters were found to be similar to each other with no statistical differences (p > .05).     

Biocontrol of saprolegniosis in rainbow trout (Oncorhynchus mykiss Walbaum) using two bacterial isolates (LE89 and LE141) of Pseudomonas fluorescens

The probiotic activity of 15 bacterial isolates that inhibit Saprolegnia parasitica in vitro was tested for the biocontrol of saprolegniosis in rainbow trout (Oncorhynchus mykiss Walbaum), adding the bacteria to tank water for 14 days at a concentration of 10⁶ bacteria ml^?1 water. Pseudomonas fluorescens LE89 and Pseudomonas fluorescens LE141 were effective in controlling experimental infection with S. parasitica since of the fish treated with LE89, 24.5% ± 16.27% (p < 0.05) became infected, as did 42.8% ± 8.41% (p < 0.05) of those treated with LE141. Given their protective effect when administered in water, their effect was also studied when administered in feed before and after experimental infection. Both bacterial isolates survived low pH levels and the action of bile, grew in skin and intestinal mucus, were resistant to several antibiotics and survived in feed; however, neither of the two isolates prevented S. parasitica infection when administered in feed.     

Screening of a high bioflocculant‐producing bacterial strain from an intensive fish pond and comparison of the bioflocculation effects with Rhodococcus erythropolis

A high bioflocculant‐producing bacterial strain BX‐13 isolated from an intensive fish pond by multiple subcultures and measured through primary screening and rescreening using kaolin. The strain BX‐13 was identified to belong to the same branch as Bacillus subtilis by morphological, physiological and biochemical, 16S rDNA gene sequencing, Blast homology search and multiple sequence alignments. The bioflocculation potential of BX‐13 and Rhodococcus erythropolis was compared, using water from the intensive pond where BX‐13 was isolated. The results showed the number of colonies in the BX‐13 treatment was higher than that of the R. erythropolis treatment from day 10 to the end of the experiment (p < 0.05). The numbers of both BX‐13 and R. erythropolis in the water body peaked on day 15, with 2.23 × 10⁷ cfu/L and 9.90 × 10⁶ cfu/L respectively. The bioflocculation volume (BFV) in water increased and then stabilized in both BX‐13 and R. erythropolis treatments. The BFV in BX‐13 peaked on day 25 (52.13 ml/L) and R. erythropolis peaked on day 30 (22.63 ml/L). At the end of the experiment, the BFV of the BX‐13 treatment was higher than the R. erythropolis treatment (p < 0.05). In conclusion, B. subtilis BX‐13 has a very strong flocculation effect, and merits further research into its application for water quality regulation in aquaculture systems.     

Effects of dietary n‐3HUFA on juvenile white shrimp,Litopenaeus vannamei: Growth,feed utilization,antioxidant enzymes activities and fatty acid compositions

Present study investigates the effects of n‐3 high‐unsaturated fatty acid (n‐3HUFA) levels on growth performance, antioxidant enzymes activities and fatty acid compositions of juvenile Litopenaeus vannamei. These represented seven iso‐nitrogenous and iso‐lipidic diets. Analysed n‐3HUFA concentrations were 0.16% (control), 0.48%, 0.74%, 1.39%, 2.39%, 2.92% and 3.44% respectively. A total of 840 juvenile L. vannamei were randomly stocked into 21 0.5 m³ tanks for 56 days. A significant increase (p < 0.05) was observed from 0.16% to 0.74% n‐3HUFA and a decrease when n‐3HUFA was above these levels in weight gain rate (WGR) and specific growth rate. Total cholesterol, triglyceride and low‐density lipoprotein in serum showed a significant decrease, high‐density lipoprotein showed a significant increase (p < 0.05). Phenoloxidase activity in serum and sodium‐potassium adenosine triphosphatase activity in gill were significantly affected by dietary n‐3HUFA (p < 0.05), both of them showing a downward trend after upward. Malic dehydrogenase and superoxide dismutase activities in serum were also significantly affected by dietary n‐3HUFA (p < 0.05), which rose first and then decreased in general, both of them have a maximum in 2.39% group. No significant differences of the activities of aspartate transaminase and alanine transaminase were observed among all groups (p > 0.05). With dietary n‐3HUFA increase, both ∑HUFA and n‐3HUFA contents gradually increased in hepatopancreas and tail muscles (p < 0.05). Based on broken‐line regression analysis of WGR, the optimal n‐3HUFA requirement is 0.89% for juvenile L. vannamei with initial weight of 0.50 ± 0.01 g.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

Effects of dietary dl‐methionyl‐dl‐methionine (Met‐Met) on growth performance,body composition and haematological parameters of white shrimp (Litopenaeus vannamei) fed with plant protein–based diets

A 10‐week feeding trial was conducted to evaluate the effects of supplementing different levels of dl ‐methionyl‐dl ‐methionine (AQUAVI^® Met‐Met) in plant protein–based diets on Litopenaeus vannamei. The positive control (PC) and negative control (NC) diets were designed with 20% and 8% fishmeal respectively, and other six diets were formulated with graded levels of Met‐Met from 0.05% to 0.30% with a 0.05% increment on the basis of NC diet (MM 0.05–MM 0.3). Six replicates were randomly assigned to each diet with 50 shrimp each having initial weight of 0.98 ± 0.02 g. The variation of FM concentration from 20% to 8% and supplemented with graded levels of Met‐Met did not affect the survival rate, feed conversion ratio, protein efficiency ratio, whole body and muscle proximate compositions (p > 0.05). However, diets with ≤0.20% Met‐Met supplementation resulted in significantly increased weight gain and specific growth rate, after which both parameters reached plateau. Shrimp fed the NC diet showed significantly lower total essential amino acid (EAA) content in muscle (p < 0.05). Supplementation of Met‐Met significantly improved apparent digestibility coefficients of dry matter, crude protein, lipid, phosphorus and EAAs (p < 0.05). Based on broken‐line analysis, the methionine requirement for white shrimp was estimated to be 0.87% when using Met‐Met as methionine source.