Effects of dietary astaxanthin on growth,antioxidant capacity and gene expression in Pacific white shrimp Litopenaeus vannamei

Pacific white shrimp Litopenaeus vannamei (1050 individuals with initial weight of 1.01 ± 0.001 g) were fed either control diet or one of six dietary astaxanthin (AX) concentration (25, 50, 75, 100, 125 and 150 mg kg⁻¹) diets for 56 days in 35 tanks (30 shrimp per tank). After 56 days of culture, shrimp‐fed AX125 and AX150 diets had higher (P < 0.05) weight gain, specific growth rate, total antioxidant status and lower (P < 0.05) superoxide dismutase (SOD), catalase (CAT) than shrimp fed control diet. After low dissolved oxygen stress for 1 h, survival rate of shrimp fed AX75, AX100, AX125 and AX150 diets was higher (P < 0.05) than that of shrimp fed control diet. Hypoxia inducible factor‐1α (HIF‐1α), cytosolic manganese superoxide dismutase (cMnSOD) and CAT mRNA expression levels of shrimp fed seven diets were significantly down‐regulated under hypoxia than under normoxia, but their expression levels were higher under hypoxia in shrimp fed AX‐supplemented diets than in shrimp fed control diet. About 70‐kDa heat‐shock protein (Hsp70) mRNA expression level of shrimp fed seven diets was significantly up‐regulated under hypoxia than under normoxia, but its expression level was lower under hypoxia in shrimp fed AX‐supplemented diets than in shrimp fed control diet.     

Evaluation of a novel bacterial biomass as a substitution for soybean meal in plant‐based practical diets for Pacific white shrimp Litopenaeus vannamei

Three growth trials and a digestibility trial were designed to evaluate the efficacy of a novel bacterial biomass (BB) in commercial‐type feed formulation for Pacific white shrimp, Litopenaeus vannamei. In trial 1, the basal diet was supplemented with 0, 60 and 120 g/kg BB to replace soybean meal (SBM). Significant improvement was observed in the survival when BB was incorporated in the diets. However, shrimp fed diets containing 120 g/kg BB exhibited significantly lower weight gain (WG) and higher feed conversion ratio (FCR). To confirm the results from trial 1 and explore the effects of BB supplementation at low levels, the basal diet was incorporated with 0, 10, 20, 40, 60 and 120 g/kg BB to replace SBM in trial 2. Significant reductions in WG, FCR, lipid content of whole body, protein retention efficiency and most amino acids retention efficiency were detected in shrimp fed with diet containing 120 g/kg BB. Trial 3 was designed to elucidate whether the digestible protein is the cause of reduced growth. No improvements in terms of growth performance and FCR were detected in the treatments balanced for digestible protein. Apparent digestibility coefficients of energy, protein and amino acid (AA) for BB were determined using chromic oxide as an inert marker and the 70:30 replacement technique. The energy, protein and individual amino acid digestibility coefficients of BB were significantly lower than those of fish meal (FM) and SBM that were given at the same time. Results of this study indicated that BB can be utilized up to 40 g/kg in shrimp feed without causing a decrease in growth. However, supplementations (≥60 g/kg) of BB can result in negative effects on growth response, FCR and protein as well as amino acids retention efficiency. At the lower levels of inclusion, shrimp performance was improved when BB was supplemented on a digestibility basis; however, at the higher level of inclusion, there was no improvement, indicating there may be other nutrients limiting. Based on enhanced survival in the treatment with BB supplementation in trial 1, further research regarding the immune effects of BB in practical shrimp feed will be necessary.     

Minimum water exchange spares the requirement for dietary methionine for juvenile Litopenaeus vannamei reared under intensive outdoor conditions

We examined if minimum water exchange could spare dietary methionine (Met) required for maximum growth performance of juvenile Litopenaeus vannamei reared in an intensive outdoor system. Shrimp of 1.98 ± 0.13 g were stocked at 70 animals/m² and reared for 72 days in 50 tanks of 1 m³ under flow‐through (14.4% a day) and static (1.4%–2.9% a day) green‐water conditions at 32.0 ± 3.7 g/L salinity. Five diets with a minimum inclusion of fishmeal supplemented with a dipeptide, dl ‐methionyl‐dl ‐methionine, were formulated to contain increasing levels of Met, 4.8, 6.2, 7.2, 8.1 or 9.4 g/kg (on a dry matter basis). Each of the five diets were fed four times daily to five replicate groups. Dietary Met and water exchange significantly influenced shrimp survival, gained yield, apparent feed intake, food conversion ratio and final body weight (p < .05). Raising shrimp under limited water exchange, i.e., static versus flow‐through spared the dependence on higher levels of dietary Met to maximize shrimp body weight, from 9.4 g/kg to 8.0 g/kg (14.0 and 12.6 g/kg Met+Cys respectively). In an intensive rearing system, a reduction in water exchange is desirable as it leads to a lower need for supplemental dietary Met.     

Anti‐PirA‐like toxin immunoglobulin (IgY) in feeds passively immunizes shrimp against acute hepatopancreatic necrosis disease

Acute hepatopancreatic necrosis disease (AHPND), caused by a toxin‐producing Vibrio parahaemolyticus strain, has become a serious threat to shrimp aquaculture. The need to regulate antibiotic use prompted the development of alternative ways to treat infections in aquaculture including the use of chicken egg yolk immunoglobulin (IgY) for passive immunization. This study evaluated the protective effect of IgY against AHPND infection in Litopenaeus vannamei (Boone). IgY was isolated from eggs laid by hens immunized with recombinant PirA‐like (rPirA) and PirB‐like (rPirB) toxins. Whole‐egg powders having IgY specific to rPirA (anti‐PirA‐IgY) and rPirB (anti‐PirB‐IgY) and IgY from non‐immunized hen (control‐IgY) were mixed with basal diets at 20% concentrations and used to prefeed shrimp 3 days before the bacterial challenge test. Survival rates of the challenged shrimp fed the anti‐PirA‐IgY, anti‐PirB‐IgY and control‐IgY diets were 86%, 14% and 0%, respectively. Only the feed containing anti‐PirA‐IgY protected shrimp against AHPND. Increasing the concentration of rPirA antigen to immunize hens and lowering the amount of egg powder in feeds to 10% consistently showed higher survival rates in shrimp fed with anti‐PirA‐IgY (87%) compared with the control (12%). These results confirm that addition of anti‐PirA‐IgY in feeds could be an effective prophylactic method against AHPND infection in shrimp.     

Evaluating the effects of dietary resistant starch on growth,digestion, metabolism,immunity and resistance to ammonia stress in Litopenaeus vannamei

Resistant starch (RS) is considered to be a good alternative to antibiotics. A 56‐day feeding trial was followed by induction of acute ammonia stress to evaluate the effects of dietary RS on growth, digestion, metabolism, immunity and resistance to ammonia stress in Litopenaeus vannamei. The four diets used differed only in RS content: 0 g/kg (Control), 10 g/kg (RS1), 30 g/kg (RS2) and 50 g/kg (RS3). The results showed that dietary RS improved the growth and the survival rates of shrimp exposed to ammonia stress. At 56 days, the activity of digestive and metabolic enzymes (amylase, lipase, pepsin, hexokinase and pyruvate kinase) was increased in the three RS groups, while trypsin activity only increased in the RS1 and RS2 groups; immune‐related parameters (the total antioxidant capacity; the activity of superoxide dismutase, total nitric oxide synthase and phenoloxidase; and the expression of heat‐shock protein 70, thioredoxin, prophenoloxidase, lysozyme and toll gene) were increased in the three RS groups. After exposure to ammonia stress, all the selected immune parameters of the three RS groups were higher than those of the control group at 24 hr. Therefore, dietary RS improved the growth, digestion, metabolism, immunity and resistance to ammonia stress in L. vannamei.     

Effects of dietary marine microalgae,Tetraselmis suecica,on production,gene expression,protein markers and bacterial count of Pacific white shrimp Litopenaeus vannamei

The marine flagellated Chlorophyta Tetraselmis suecica is among the most important live food species in marine aquaculture. In the present study, the effects of dietary supplementation of dried marine microalgae, Tetraselmis suecica, on growth performance; feed utilization; chemical composition; gene expression of superoxide dismutase (SOD), glutathione peroxidase (GPx) and insulin‐like growth factor 2 (IGF‐II) gene of Pacific white shrimp, Litopenaeus vannamei; muscle protein polymorphism; and microbial count were assessed and evaluated. Three hundred and sixty L. vannamei (postlarvae) Pls (0.124 ± 0.002 g) were randomly stocked into 40‐L glass aquaria (30 shrimp/aquarium) and fed three times daily four tested diets: a basal diet (control), diet incorporated with 2.5 g kg^?1 dried T. suecica (T1), 5 g kg^?1 dried T. suecica (T2) and 7.5 g kg^?1 dried T. suecica (T3) in triplicates, for 90 days. At the end of the trial, the survival rate (SR) of L. vannamei fed diets supplemented with different levels of T. suecica was significantly (p < .05) higher than the control diet. The highest weight gain and specific growth rate and the best feed conversion ratio were recorded on L. vannamei fed a diet supplemented with a 7.5 g/kg dried T. suecica. The highest protein, lipid and ash contents were obtained in L. vannamei fed the diet containing 7.5 g/kg T. suecica, when compared with the remaining tested diets. The gene expression of antioxidant genes SOD and GPx was the lowest in the T3 group in comparison with the control group. Meanwhile, expression level of IGF‐II was higher in the T2 group. The total heterotrophic bacterial count was significantly (p < .05) increased with the cumulative T. suecica level, while no significant (p > .05) differences were found in the total Vibrio count among treatments. Overall, the present results have shown that the diet supplemented with the highest inclusion level of dried T. suecica resulted in improved growth and nutrient utilization.     

Proteome modifications of Pacific white shrimp (Litopenaeus vannamei) muscle under biofloc system

The objective of the study was to examine the effects of biofloc technology on the muscle proteome of Litopenaeus vannamei. Two biofloc treatments and one control were compared: biofloc‐based tanks under zero‐water exchange fed with 150 g/kg crude protein (BF15), or with 250 g/kg crude protein (BF25) diets, and clear water tanks with 50% of daily water exchange stocked with shrimp fed with similar amount of a 250 g/kg crude protein diet, referred to as control. The shrimp (5.28 ± 0.42 g) were divided into the 300‐L fibreglass tanks (water volume of 200 L) at a density of 35 shrimp per tank and were cultured for 35 days. The biofloc groups displayed better growth and survival compared to the control. The muscle tissue from the control and BF25 groups was subjected to proteomic analysis. Lactate dehydrogenase, enolase, arginine kinase, mitochondrial ATP synthase subunit alpha, mitochondrial ATPase inhibitor factor 1 precursor, serpin 3 and myeloid differentiation factor 88 had an increased abundance in the BF25 group, while myosin heavy chain type 1 and myosin heavy chain type 2 showed a decreased abundance. The results indicate that biofloc technology could alter the expression of proteins involved in structure, metabolism and immune status of cultured shrimp.     

The effects of dietary Eucommia ulmoides Oliver on growth,feed utilization,antioxidant activity and immune responses of turbot (Scophthalmus maximus L.)

The present study was conducted to evaluate the effects of dietary Eucommia ulmoides (EU) on growth, feed utilization, antioxidant activity and immune responses of turbot fed a basal diet (CON) or EU‐supplemented diets with 5.0 g/kg (EU1), 10.0 g/kg (EU2) and 20.0 g/kg (EU3) EU leaf powder. After 70‐day trial, EU supplementation did not affect nutrient utilization, but reduced feed intake (FI) and specific growth rate (SGR) of fish at doses above 5.0 g/kg. Superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T‐AOC) activities in the EU‐supplemented groups were significantly higher than those in the control group at a dose of 20.0 g/kg. Significantly reduced malondialdehyde (MDA) contents were observed in EU‐supplemented groups at doses over 5.0 g/kg. Furthermore, fish fed 20.0 g/kg EU showed the highest lysozyme (LZM) activity among groups. The EU‐supplemented diets with doses above 5.0 g/kg significantly enhanced the mRNA expressions of cytokines. The expression level of major histocompatibility complex II alpha (MHC IIα) was significantly upregulated compared to that of the control fish when the supplemental level was at 20.0 g/kg. Taken together, the present study indicated that the EU could remarkably enhance the antioxidant activity, non‐specific immunity and maintain an active immune response in turbot.     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.     

Hypoxia‐induced changes in survival,immune response and antioxidant status of the Pacific white shrimp (Litopenaeus vannamei) fed with graded levels of dietary myo‐inositol

A 3‐hr experiment was conducted to investigate the effects of dietary myo‐inositol (MI) supplementation on survival, immune response and antioxidant abilities in Litopenaeus vannamei under acute hypoxia stress. Six practical diets were formulated with supplementation of graded levels (control group 0, 0.1, 0.2, 0.4, 0.8 and 1.6 g/kg dry diet) of MI and were randomly assigned to triplicate groups of L. vannamei (mean weight 0.40 ± 0.00 g) for 8 weeks. Ten healthy shrimp (final mean weight approximately 11–14 g) randomly selected from each tank were exposed to hypoxia stress after feeding trial. After 3‐hr acute hypoxia stress, survival of shrimp fed MI‐supplemented diets (except 0.1 and 0.4 g/kg diets) was significantly increased compared with the control group. Shrimp fed control diet had lower activities of alkaline phosphatase (AKP), acid phosphatase (ACP), total antioxidant capacity (T‐AOC) and glutathione peroxidase (GPX), and higher malondialdehyde (MDA) and protein carbonyl (PC) contents in hepatopancreas than those fed the MI‐supplemented diets. In addition, mRNA expression levels of heat shock protein 70 (Hsp70), catalase (CAT) and penaeidin were significantly differentially regulated in hepatopancreas. In summary, dietary MI supplementation may have a positive effect on improving resistance to acute hypoxia stress of L. vannamei.     

Administration of probiotic Bacillus licheniformis induces growth,immune and antioxidant enzyme activities,gut microbiota assembly and resistance to Vibrio parahaemolyticus in Litopenaeus vannamei

This study aimed to elucidate the effects of dietary Bacillus licheniformis ATCC 11946 (BL) on the growth, immune and antioxidant activities, intestinal morphology and microbiota, and susceptibility to Vibrio parahaemolyticus (VP_AHPND) in juvenile pacific white shrimp, Litopenaeus vannamei (LV). Juvenile LV (initial weight = 0.63 ± 0.001) were fed diets containing varying BL concentrations (0 (BL0), 10⁶ (BL1), 10⁷ (BL2) and 10⁸ (BL3) CFU/g feed) for 8 weeks. Growth performance, immune and antioxidant enzyme activities, and intestinal morphology significantly improved in the probiotic‐treated groups than the untreated. Regardless of the treatment group, the two dominant phyla were Proteobacteria and Bacteroidetes, whereas the two dominant genera were Ruegeria and Vibrio. Increasing inclusion of probiotics in diets led to significant increase in beneficial bacterial genera (Ruegeria, and Pseudoalteromonas) and a significant decrease in some known opportunistic pathogens (Vibrio, Tenacibaculum, Photobacterium, Kangiella and Spongiimonas) with the BL3 group witnessing the best. A 7‐day challenge study with VP_AHPND showed significantly high protection in the probiotic‐treated groups, with the BL3, BL2 and BL1 obtaining 51%, 34% and 22% relative percentage survival, respectively. In conclusion, BL at 10⁸ CFU/g feed should be used to help in shrimp production since it attained the most significant improvement.     

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

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

Effects of fructooligosaccharide on growth,immunity and intestinal microbiota of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

The effects of fructooligosaccharide (FOS) on growth performance, immunity and predominant autochthonous intestinal microbiota of shrimp (Litopenaeus vannamei) fed diets with fish meal (FM) partially replaced by soybean meal (SBM) were evaluated. After acclimation, shrimps (1.82 ± 0.01 g/kg) were allocated into 15 tanks (25 shrimps per tank) and fed five different diets including positive control diet (C0, containing 250 g/kg FM and 285 g/kg SBM), control diet (C, containing 125 g/kg FM, 439 g/kg SBM) and three experimental diets supplemented with 1.0 g/kg FOS (T1), 2.0 g/kg FOS (T2) and 4.0 g/kg FOS (T3) to control diet (C) respectively. Shrimps were fed diets to apparent satiation three times per day, and 15 shrimps from each aquarium were randomly sampled and analysed at the end of the 6‐week feeding trial. The results showed that FBW, WGR, SGR and SR decreased, while FCR and FI increased significantly in control (C) compared with positive control (C0). Besides, significantly decreased trypsase and lipase activities, and SOD, AKP and ACP activities were recorded in control (C) compared with positive control (C0). On the other hand, significantly improved SGR and decreased FCR were observed in groups T1, T2 and T3 compared with control (C). Moreover, lipase and amylase activities enhanced significantly in group T3 compared with the control (C), while GOT and GPT activities dropped significantly with the increment supplementation of FOS in diets. Compared with the control (C), SOD activity enhanced significantly and MDA level decreased significantly in groups T2 and T3, and improved AKP and ACP activities were observed in group T3. In addition, dietary FOS improved the microbial diversity, and suppressed several potential pathogens, such as Vibrio tubiashii, Vibrio parahaemolyticus and Photobacterium damselae‐like strains in the intestine of shrimp. Overall, these results proved FOS could relieve the side effects induced by SBM and supported the use of 2.0–4.0 g/kg FOS in shrimp diets with FM partially replaced by SBM.     

Dietary nucleotide‐rich yeast supplementation improves growth,innate immunity and intestinal morphology of Pacific white shrimp (Litopenaeus vannamei)

An 8‐week feeding trial was conducted to evaluate the effects of dietary nucleotide (NT)‐rich yeast supplementation on growth, innate immunity and intestinal morphology in Pacific white shrimp (Litopenaeus vannamei). Four isonitrogenous and isolipidic practical diets were formulated to contain 0 (control), 10, 30 and 50 g/kg of NT‐rich yeast, respectively. A total of 480 shrimp with an average initial body weight of 1.86 ± 0.02 g were randomly allocated into four groups, with four replicates per group and 30 shrimp each replicate. The results indicated that shrimp fed the diet containing 50 g/kg NT‐rich yeast had significantly higher weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) than those fed the control diet, and the lowest feed conversion ratio (FCR) was observed in the shrimp fed the 50 g/kg NT‐rich yeast supplemental diet. However, there was no significant difference in survival among all treatments. The crude protein of whole shrimp in the 50 g/kg NT‐rich yeast group was higher than that in the control group. Total protein, triglyceride concentrations, the activities of aspartate aminotransferase and alanine aminotransferase in serum were significantly influenced by the dietary NT‐rich yeast supplementation. The activities of serum phenoloxidase (PO) and lysozyme (LZM) of shrimp fed the diet containing 50 g/kg NT‐rich yeast were higher than those in shrimp fed the other diets. Relative expressions of alp and lzm significantly upregulated in the 30 g/kg NT‐rich yeast group compared to the control group. The intestinal fold height and fold width in the 30 g/kg NT‐rich yeast group were significantly higher than those fed the control diet; and the highest microvillus height occurred in the shrimp fed the 50 g/kg NT‐rich yeast diet. In summary, dietary 30–50 g/kg NT‐rich yeast supplementation promotes growth performance, enhances innate immunity and improves intestinal morphology of Litopenaeus vannamei.     

Evaluation of dried fermented biomass as a feed ingredient in plant‐based practical diets for juvenile Pacific white shrimp Litopenaeus vannamei

Two growth trials were designed to evaluate the utilization of dried fermented biomass (DFB) in commercial type feed formulation for Pacific white shrimp, Litopenaeus vannamei. In trial 1, four experimental diets were formulated to utilize increasing levels (0, 25, 50 and 100 g/kg) of spray‐dried fermented biomass (SDFB) as a replacement of fish meal (FM). Results indicated that SDFB can be utilized up to 50 g/kg as a substitution for FM without causing growth depression in shrimp. However, dietary SDFB supplementation at 100 g/kg significantly reduced the weight gain (WG) of shrimp and increased feed conversion ratio (FCR). This reduction in performance is likely due to palatability or nutrient imbalances of the feed. In trial 2, nine experimental diets were formulated with increasing levels (0, 20, 40, 60 and 120 g/kg) of spray‐dried (S) or granular (G) DFB to replace soy protein concentrate (SPC) or SPC + corn protein concentrate (CPC). This allowed the comparison between spray‐dried and ring‐dried products. Ring drying produced a granular product, reducing dust and increasing product particle size. Shrimp fed with diet containing 20 g/kg GDFB performed the best in terms of final mean weight, WG and FCR. Significantly reduced growth and increased FCR were observed in shrimp fed diets containing 60 and 120 g/kg SDFB. Lipid content of whole body was significantly reduced when GDFB was incorporated at 120 g/kg. No significant differences were detected in survival, protein retention efficiency as well as protein and ash contents of the whole shrimp. Results from analysis of covariance indicated that the processing method (covariant) had a significant effect on final mean weight, WG and FCR. In general, shrimp fed with diet containing granular product performed better as compared to those fed with diets utilizing spray‐dried product. GDFB can be utilized in the diets up to 120 g/kg in practical shrimp feeds as a substitute for SPC and CPC without compromising the growth of shrimp. However, a significant reduction in WG was observed in the diets containing 60 and 120 g/kg SDFB. The results in the current study demonstrate that processing changes to produce a granular product produced an improved feed ingredient for shrimp.     

Strain and dose infectivity of Vibrio parahaemolyticus: the causative agent of early mortality syndrome in shrimp

Diseases of shrimp have contributed to billions of dollars of economic loss in the aquaculture industry. Newly emerging strains of the bacterium Vibrio parahaemolyticus produce a condition in shrimp called early mortality syndrome or acute hepatopancreatic necrosis disease. Three different V. parahaemolyticus strains were evaluated for their respective pathogenicity on shrimp, Litopenaeus vannamei, when the bacterial strains were grown under various laboratory conditions prior to inoculating shrimp. For each trial, feed was inoculated with a known concentration of bacteria and then fed to the shrimp. The early mortality syndrome strain of V. parahaemolyticus was the most lethal resulting up to 100% mortality within 24 h after being introduced to shrimp via a single feeding. The other two strains of Vibrio, one isolated from the environment and the other from a human clinical case, resulted in 0% and 30% mortality within 96 h respectively. The concentration of the early mortality syndrome strain of V. parahaemolyticus that the shrimp were exposed to directly correlated with mortality rate, which allowed for lethal or sublethal short‐term disease challenge assays to be established. Infiltration of haemocytes was also evident in the midgut caeca of shrimp infected with the early mortality syndrome strain of V. parahaemolyticus, which has not been previously reported.     

Growth performance,immune response and disease resistance of rainbow trout (Oncorhynchus mykiss) fed Thymus vulgaris essential oils

Efficacy of Thymus vulgaris essential oils was assessed on growth, immune response and disease resistance of rainbow trout (Oncorhynchus mykiss). Fish weighing 10 g were fed with dietary supplemented of the oils at 0.5, 1.0 and 2.0 ml/kg feed for 2 months. Fish fed with the oils at 0.5 ml/kg feed demonstrated a better weight gain and specific growth rate, compared to other treatments (p < .05). Fish fed with 1.0 ml the oils showed the highest up‐regulation of complement component 3 (C3) and (cluster of differentiation 4) (CD4) genes expression (p < .05), while lysozyme gene expression level significantly increased in fish fed with 2.0 ml of the oils. In addition, at the end of the experiment, the expression of C3 and CD4 genes were significantly up‐regulated in fish fed with 1.0 and 2.0 ml of the oils, while IL‐1ß and lysozyme genes expression levels were significantly decreased in fish fed 2.0 ml oils, towards the end of the trial (p < .05). There was a fluctuation in the levels of Alanine aminotransferase, Aspartate aminotransferase and Alkaline phosphatase in all treatments during the experiment. When treated fish were challenged with Aeromonas hydrophila, the highest survival rate was observed in 0.5 ml/kg treatment, followed by 2 and 1 mg/kg treatments. Overall, these findings demonstrated that dietary administration of T. vulgaris oils especially at 0.5 mg/kg feed can be considered as a potential component for enhancing of the growth, immune responses and disease resistance of trout against motile Aeromonas septicemia caused by A. hydrophila.     

Fungus,Aspergillus niger,fermented groundnut oil cake as a fishmeal alternative in the diet of Penaeus vannamei

Commercial de‐fatted groundnut oil cake (GNC) fermented with the fungus Aspergillus niger was evaluated as a fishmeal alternative in the diet of Penaeus vannamei. A 45‐day growth trail was performed using nine iso‐nitrogenous and iso‐lipidic diets. Untreated/fermented GNC was included at the rate of 0 (control), 25, 50, 75 and 100 g/kg by replacing fishmeal (w/w). Each diet was randomly assigned to triplicate group of 20 shrimps (initial weight of 3.09 ± 0.03 g). Results revealed that shrimp fed with diets having untreated GNC up to 50 g/kg has no significant difference in growth, whereas the inclusion level was enhanced to 100 g/kg with fermented GNC with no deleterious effect. The negative linear trend was found for SGR with increasing the inclusion of both untreated and fermented GNC. The feed and protein efficiency measures, viz. feed conversion ratio, protein efficiency ratio and apparent protein utilization, were better in shrimps fed with diets having fermented GNC than those fed the respective level of untreated GNC. The broken‐line analysis indicated that the inclusion of 72.5 g/kg fermented GNC showed the best FCR. No significant difference was observed in survival (86.67%–96.67%) between the dietary treatments. There was a significant difference in ether extract of shrimp between the treatments, whereas other parameters were not affected. Haemolymph indices showed a significant difference in total protein, glucose, cholesterol and triglycerides between control and test diets. The results conclude fermented GNC, which can be better than untreated one in the diet of shrimp.     

Effect of lignite fulvic acid on growth,antioxidant ability,and HSP70 of Pacific white shrimp, <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

This study was conducted to evaluate the effects of fulvic acid (0, 0.3, 0.6, 0.9, and 1.2%) as feed additive on growth, feed utilization, antioxidant ability, and HSP70 in hemolymph and hepatopancreas of juvenile white shrimp Litopenaeus vannamei (average weight 2.5 g) reared under experiment conditions. Shrimp were stocked at a density of 625 shrimps m^?3 for 60 days in net cages submerged in recirculating tanks. At the end of the experiment, specific growth rates and survival rates of shrimp in treatment groups fed with 0.6, 0.9, and 1.2% fulvic acid were higher compared to that of the control group. Shrimp fed 0.6, 0.9, and 1.2% fulvic acid had significantly lower feed conversion rates than those fed control diet. The optimum dietary fulvic acid requirement for juvenile shrimp based on weight gain was 0.897%. Furthermore, superoxide dismutase activity and peroxidase activity increased significantly, while malonaldehyde content decreased in the hemolymph and hepatopancreas of shrimp fed 0.9 and 1.2% dietary fulvic acid. Glutathione content increased obviously in hemolymph of shrimp fed 0.6, 0.9, and 1.2% fulvic acid. In hepatopancreas, glutathione content was significantly higher in shrimp supplemented with 1.2% fulvic acid. HSP70 decreased obviously in hemolymph of shrimp fed 0.9 and 1.2% fulvic acid, while shrimp fed with 0.6 and 0.9% fulvic acid showed lower HSP70 level in hepatopancreas. The results of this study demonstrated that dietary fulvic acid could improve survival rates, growth, feed utilization, antioxidant capability, and stress resistance of juvenile L. vannamei reared under intensive stocking conditions.