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.     

Genetic parameters and response to selection for harvest body weight of pacific white shrimp,Litopenaeus vannamei

Genetic parameters and response to selection were estimated for the harvest body weight of Litopenaeus vannamei. The data consisted of 24 072 progeny from 178 sires and 171 dams in two generations (G₀ and G₁) with a nested mating structure. All families were randomly divided into two groups and then cultured in two farms at different locations (Huanghua and Qingdao). The heritability estimates from G₀ and G₁ were 0.278 ± 0.136 and 0.423 ± 0.065 respectively. Over two generations, the heritability estimate was 0.335 ± 0.087, and the common environmental effect was 0.084 ± 0.031. A bivariate animal model was used to estimate variance and covariance components, whereby the body weight in the two farms was treated as a genetically distinct trait. Genetic correlation was close to unity (0.943 ± 0.066), indicating that a genotype by environmental interaction for harvest body weight was small. The response to selection in harvest body weight was estimated using two methods (the realized and predicted responses). The realized response was estimated from the difference in the least squares means of body weight for the selection and control populations. The predicted response was obtained from the difference in the mean estimated breeding values between generations. The realized response was 2.30%, while the predicted responses were 2.00% and 1.37% for within‐ and across‐generation datasets using two sets of genetic parameters respectively. The results would provide crucial information in pacific white shrimp breeding programs in China.     

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

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

Efficacy of crystalline methionine and microencapsulation methionine in diets for Pacific white shrimp Litopenaeus vannamei

An 8‐week growth trial was conducted to evaluate the efficacy of crystalline methionine (C‐Met) or microencapsulated methionine (M‐Met) in practical diets for Pacific white shrimp Litopenaeus vannamei. A high fishmeal reference diet was formulated with 15% fishmeal, and then, soybean meal (SBM) was used to replace 50% fishmeal as a low fishmeal basal diet (7.5% fishmeal). Graded levels (0.1%, 0.15% and 0.2%) of methionine originating from C‐Met or M‐Met were added to the basal diet. Each diet was randomly assigned to three tanks (40 shrimps per tank) in an indoor flow through seawater system. The results showed that the shrimp fed the basal diet with 0.15% or 0.20% methionine originating from 0.375% or 0.5% M‐Met had significantly higher final weight and weight gain as compared to shrimp fed the basal diet. The supplementation M‐Met in the basal diets resulted in increased serum ammonia in shrimp. Hepatopancreas amylase activity of shrimp showed a decreasing trend with increasing methionine supplementation, which is in contrast with the upswing trend in trypsin at pancreatic segment. This study indicated that the diets supplemented with 0.15% or 0.20% of methionine from M‐Met (0.58% or 0.65% of dietary methionine respectively) were effective in improving the nutritional value of SBM‐based diets deficient in methionine (0.48%) for the Pacific white shrimp.     

Genetic variation of white spot syndrome virus (WSSV) in Pacific white shrimp Litopenaeus vannamei (Boone 1931) culture of Thailand

White spot syndrome virus (WSSV) is highly pathogenic to penaeid shrimp and has caused significant economic losses in the shrimp farming industry in Thailand. Genotyping analysis was done in 124 WSSV isolates from cultured Pacific white shrimp Litopenaeus vannamei. These samples were obtained during 2007–2014 from eight provinces in Thailand. We investigated five variable loci in the virus genome: deletions in two variable regions, VR14/15 and VR23/24, and three variable number tandem repeats (VNTR) located in open reading frame (ORF) 75, 125 and 94. WSSV genotype was characterized as (X_14/15, X_23/24) (N₇₅‐N₁₂₅‐N₉₄) where X is the number of base pair deletion in the variable region and N is the number of repeat units (RUs) in a specific ORF. The deletion pattern in VR14/15 and VR23/24 regions characterized three WSSV genotypes. The most prevalent genotype was (5950_14/15, 10971_23/24), and it was found in all studied areas. At least 33 genotypes of WSSV were analysed based on 3 VNTR loci, indicating that the VNTRs of WSSV genome are highly variable. From 124 WSSV samples, two samples presented the characteristic of all five variable loci similar to WSSV collected during 2010 in Saudi Arabia (5950_14/15, 10971_23/24) (3₇₅‐6₁₂₅‐7₉₄). Many different WSSV genotypes shown in this study as compared to previously reported genotypes in Thailand suggests current status of disease epidemiology, as well as probable movements of WSSV between countries.     

Dietary thiamin could improve growth performance,feed utilization and non‐specific immune response for juvenile Pacific white shrimp (Litopenaeus vannamei)

This study evaluated the effect of dietary thiamin on growth performance, feed utilization and non‐specific immune response for juvenile Pacific white shrimp, Litopenaeus vannamei. Six isonitrogenous and isolipidic practical diets were formulated with graded thiamin levels of 6.9, 32.7, 54.2, 78.1, 145.1 and 301.5 mg kg^?1 of dry diet, respectively. Each diet was randomly assigned to triplicate groups of 30 juvenile shrimp and provided four times each day to apparent satiation. Weight gain (WG) and specific growth rate (SGR) of the shrimp were significantly influenced by the dietary thiamin levels, the maximal WG and SGR occurred at 54.2 mg kg^?1 dietary thiamin level. However, with further increase in dietary thiamin level from 54.2 to 301.5 mg kg^?1, the WG and SGR significantly decreased. Shrimp fed the 54.2 mg kg^?1 thiamin diet exhibited higher feed efficiency, protein efficiency ratio and protein productive value than those fed the other diets. Dry matter and protein content in whole body were significantly affected by the dietary thiamin levels. Thiamin concentration in hepatopancreas significantly increased when the dietary thiamin level increased from 6.9 to 145.1 mg kg^?1. The total protein, glucose, triacylglycerol and cholesterol contents in hemolymph were not significantly affected by the dietary thiamin levels. Dietary thiamin had significantly influenced superoxide dismutase, catalase and lysozyme activities in hemolymph. Results of this study indicated that the optimal dietary thiamin requirements estimated using a two‐slope broken‐line model based on WG and thiamin concentration in hepatopancreas were 44.66 and 152.83 mg kg^?1, respectively.     

Genetic analysis of the Pacific white shrimp (Litopenaeus vannamei): heterosis and heritability for harvest body weight

The aim of this study was to estimate heterosis and heritability for harvest body weight of the Pacific white shrimp (Litopenaeus vannamei) measured at commercial farm conditions. Heterosis and heritability were estimated using a base population from diallel crosses of eight introduced strains. The base population included 9936 shrimp from 207 families that were produced with 188 sires and 172 dams using a nested mating design by artificial insemination. Heterosis was calculated basing on the least squares means (LSM) of harvest body weight. The results showed that most of the hybrids (75%) have positive heterosis for harvest body weight, which ranged from ?13.36% (UA2 × UA5) to 13.80% (UA6 × UA5) with a mean of 2.41%. The high amount of heterosis manifested in the hybrids indicated the usefulness of these hybrids for improving the growth. Variance components and heritability for harvest body weight were estimated using an animal model. The heritability estimate for harvest body weight was 0.092 ± 0.082 (h²) when genetic groups were excluded from the pedigree, but it was decreased when genetic groups were included in the pedigree ( = 0.066 ± 0.050), implying that there are strain additive genetic effect and heterosis in the base population. However, the heritability estimates for harvest body weight were significantly different from zero (P < 0.05) and there was no significant difference between h² and (P > 0.05). The results from this study indicated that significant improvement for growth is possible through cross‐breeding and selective breeding in L. vannamei.     

White spot syndrome virus epizootic in cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Taiwan

White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV‐positive farms: post‐larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7–100% nucleotide sequence identity with four strains in the GenBank database – China ( AF332093 ), Taiwan ( AF440570 and U50923 ) and Thailand ( AF369029 ). This is the first broad study of WSSV infection in L. vannamei in Taiwan.     

Effect of bioflocs on growth and immune activity of Pacific white shrimp,Litopenaeus vannamei postlarvae

The bioflocs technology (BFT) for shrimp production has been proposed as a sustainable practice capable of reducing environmental impacts and preventing pathogen introduction. The microbial community associated with BFT not only detoxifies nutrients, but also can improve feed utilization and animal growth. Biofloc system contains abundant number of bacteria of which cell wall consists of various components such as bacterial lipopolysaccharide, peptidoglycan and β‐1, 3‐glucans, and is known as stimulating nonspecific immune activity of shrimp. Bioflocs, therefore, are assumed to enhance shrimp immunity because they consume the bioflocs as additional food source. Although there are benefits for having an in situ microbial community in BFT systems, better understanding on these microorganisms, in particular molecular level, is needed. A fourteen‐day culture trial was conducted with postlarvae of Litopenaeus vannamei in the presence and absence of bioflocs. To determine mRNA expression levels of shrimp, we selected six genes (prophenoloxidase1, prophenoloxidase2, prophenoloxidase activation enzyme, serine proteinase1, masquerade‐like proteinase, and ras‐related nuclear protein) which are involved in a series of responses known as the prophenoloxidase (proPO) cascade, one of the major innate immune responses in crustaceans. Significant differences in shrimp survival and final body weights were found between the clear water and in the biofloc treatments. mRNA expression levels were significantly higher in the biofloc treatment than the clear water control. These results suggest that the presence of bioflocs in the culture medium gives positive effect on growth and immune‐related genes expression in L.vannamei postlarvae.     

Induction of spawning in Pacific white shrimp Litopenaeus vannamei (Boone, 1931) by injection of its molt inhibiting hormone isoform II produced in E. coli

The optimization of reproductive parameters in shrimp farming continues to be a challenge for most producing countries. Although the crustacean neuropeptides have been studied extensively in the last two decades, the functions of most of these neuropeptides remained putative. Among them, molt‐inhibiting hormone isoform II (MIH II) has shown an important role in vitellogenesis. In this study, the cDNA encoding mature MIH II peptide was isolated by RT‐PCR from the L. vannamei eyestalk. The cDNA was cloned into pET28a bacterial expression vector. Recombinant MIH II was obtained in the form of insoluble inclusion bodies and purified to ~88% purity. Two doses of rMIH II and a negative control group were assayed in vivo. The stages of ovarian maturation and spawning were recorded during 72 hr post‐injection. The results showed that ovarian maturation occurred approximately in 9% and 33% of females injected with rMIH II at the doses of 300 and 600 ng/gbw respectively. Neither maturation nor spawning was detected in the negative control group. Females injected with 600 ng/gbw, which showed vitellogenic stages III and IV, spawned. These preliminary results argue that the hormone rMIH II could be a promising candidate to induce spawning in L. vannamei shrimp.     

Response of Pacific white leg shrimp (Litopenaeus vannamei) on exposure to aflatoxin in feed

A research was accomplished to find the response of Litopenaeus vannamei to different levels of aflatoxin in feed. Two hundred live L. vannamei were collected from a shrimp breeding station located on the south of Tiab, in Hormozgan province. Aflatoxin contaminated diets with 0.18, 96.21, 184.74, 711.35, 977.11 and 1605.61 µg/kg were prepared and shrimps in six experimental groups with three replicates in each group for 28 days were fed with these diets. The results showed that activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) in haemolymph and histological alternation index (HAI) value of hepatopancreas in treatement groups were significantly higher than control group (p < 0.05). Decreased total protein and fat in haemolymph and increased protein in muscle of treatment shrimps were observed compared with the control group (p < 0.05). The HAI value of 977.11 and 1605.61 μg/kg groups were significantly higher than the control and other groups (p < 0.05). Histological study results showed that calculated HAI (more than 100) in these groups as a result of necrosis lesion observed in hepatopancreas. Moderate to severe alteration was recognized in group fed by 711.35 µg/kg aflatoxin. Minor aflatoxin levels (0.18, 96.21 and 184.74 µg/kg) caused inappreciable tissue alteration. This study demonstrated body composition, haemolymph parameters and histology of hepatopancreas were affected in the aflatoxin‐contaminated feed when the concentration was over 955.14 µg/kg.     

Changes in the intestinal bacterial community during the growth of white shrimp,Litopenaeus vannamei

In this study, we documented the changes in the intestinal bacterial community at four stages in Litopenaeus vannamei: 14 days postlarvae (L14) and 1‐, 2‐ and 3‐month old juveniles (J1, J2, J3), using 454 pyrosequencing techniques. The intestinal bacterial community was dominated by three bacterial phyla, Proteobacteria, Bacteroidetes and Actinobacteria at all stages. However, the relative abundance and bacterial lineages varied at the family level. The intestinal bacterial community of L14 and J1 was similar, with dominant members belonging to the Comamonadaceae of Betaproteobacteria. Conversely, bacterial members affiliated to Flavobacteriaceae of Bacteroidetes were dominant in J2 and Vibrionaceae of Gammaproteobacteria was dominant in J3. The abundance of Microbacteriaceae of Actinobacteria also fluctuated during the four stages. Bacterial members of Flavobacteriaceae and Rhodobacteraceae (Alphaproteobacteria) were present through all growth stages, and likely form the intestinal core microbiome of L. vannamei. However, they varied at the operational taxonomic unit (OTU) level through the growth stages. The intestinal bacterial community of pond‐rearing shrimp included the three main bacterial phyla identified above, and an additional group, Mycoplasmataceae of Mollicutes. Our results demonstrate that the intestinal bacterial community of L. vannamei was highly dynamic during the growth stages. Bacterial members belonging to Commamonadaceae dominated in the earlier growth stage of shrimp, possibly influenced by feeding with Artemia nauplii, but there was a shift to Flavobacteriaceae in the mid and Vibrionaceae in the late growth stages.     

Quantitative dietary leucine requirement of juvenile Pacific white shrimp,Litopenaeus vannamei (Boone) reared in low‐salinity water

The experiment was conducted to determine the leucine requirement of juvenile Pacific white shrimp Litopenaeus vannamei (Boone) in low‐salinity water (0.50–1.20 g L^?1). Six diets were formulated to contain 410 g kg^?1 crude protein with fish meal, peanut meal and precoated crystalline amino acids with different concentration of l ‐leucine (16.72, 19.60, 22.06, 24.79, 27.28 and 30.16 g kg^?1 dry diet). Each diet was randomly assigned to triplicate groups of 30 shrimps (0.38 ± 0.002 g), and the feed trial lasted for 8 weeks. The results indicated that the maximum weight gain was observed at 24.95 g kg^?1 dietary leucine group, whereas the diets containing higher leucine concentration conversely reduced the growth performance (P < 0.05). Moreover, the highest body protein content and body protein deposition and the lowest haemolymph AST and ALT activities were also found at 24.95 g kg^?1 dietary leucine group. With the increase in leucine in diets, a dose‐dependent increase was found in body lipid content and haemolymph urea concentration. The polynomial regression calculated using weight gain, feed efficiency and body protein deposition indicated that the optimal dietary leucine requirement for L. vannamei reared in low‐salinity water was 23.73 g kg^?1 leucine of dry diet, correspondingly 57.88 g kg^?1 of dietary protein.     

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.     

Use of new soybean varieties in practical diets for the Pacific white shrimp,Litopenaeus vannamei

This study was designed to evaluate the efficacy of eight sources (designated A–H) of soybean meal (SBM) which included six new non‐genetically modified soya varieties in practical feed formulation for Pacific white shrimp, Litopenaeus vannamei, using both growth and digestibility trials. A soybean meal‐based reference diet was formulated using conventional soybean meal (527 g kg^?1 diet), which was then replaced on an isonitrogenous basis with various other experimental soybean meals. In a 6‐week growth trial, shrimp in four replicate tanks per dietary treatment (10 shrimp per tank, initial weight 0.52 ± 0.04 g) were cultured in a recirculating system. There were no significant differences with respects to per cent weight gain and survival across all dietary treatments; however, final weights and feed conversion ratio (FCR) were lower in shrimp offered diet 3. Apparent digestibility coefficients for the eight (A–H) different soybean meals were determined in L. vannamei for dry matter (ADMD), gross energy (ADE) and crude protein (ADP) using 10 g kg^?1 chromic oxide as inert marker with 70 : 30 replacement techniques. Coefficients ranged from 71.3% to 88.3%, from 76.6% to 91.3% and from 93.6% to 99.8%, for ADMD, ADE and ADP, respectively. Improved digestibility values were observed in soybean C which was characterized by crude protein (471 g kg^?1), crude fat (97 g kg^?1), low cooking temperature (180 °C), higher nitrogen solubility index (689 g kg^?1) and protein dispersibility index (619 g kg^?1). This indicates that new lines of soybean meal can be used to improve digestibility coefficients in shrimp feeds.     

Apparent digestibility of animal,plant and microbial ingredients for Pacific white shrimp Litopenaeus vannamei

A digestibility trial was conducted to determine apparent digestibility coefficients of dry matter, protein, energy and amino acids of animal, plant and microbial ingredients for Pacific white shrimp, Litopenaeus vannamei. The tested ingredients included traditional soybean meal (TSBM), PepsoyGen soybean meal (PSBM), NutriVance soybean meal (NSBM), fish meal (FM), poultry meal (PM), squid hydrolysis (SQH), scallop hydrolysis (SCH), flash dried yeast (FDY), two batches of Ulva meal (UMF and UMS) and bacteria biomass (BB). A basal diet was formulated and produced along with the experimental diets which included 300 g/kg of each ingredient and 700 g/kg of the basal diet. Juvenile shrimp (initial mean weight: 12 g, six shrimp/tank, n = 3) were stocked in a recirculation system. Apparent dry matter, protein and energy digestibility coefficients ranged from ?40.11% to 78.51%, 15.17% to 97.03% and 13.33% to 82.56% among different protein sources, respectively. In general, protein and energy digestibilities in soy sources (77.6% to 97.03% and 62.77% to 82.56%, respectively) are higher than the tested animal protein (51.39% to 71.41% and 45.29% to 69.77%, respectively) and single‐cell protein sources (15.17% to 53.47% and 13.33% to 40.39%, respectively). Among the three soybean sources, TSBM showed highest protein and energy digestibility. Apparent individual amino acid digestibility coefficients were also variable among different types of ingredients, and there was a reasonable correspondence to protein digestibility. The most digestible feed ingredients for Pacific white shrimp in this study were conventional soybean meal (SBM) and NutriVance soybean meal (NSBM), which indicated that these ingredients are good protein and amino acid sources for Pacific white shrimp. Resultant digestibility data may provide useful information to commercial shrimp feed industry.     

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.     

Dietary choline improves growth performance,antioxidant ability and reduces lipid metabolites in practical diet for juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of dietary choline levels on growth performance, antioxidant capacity and lipid metabolites in juvenile Pacific white shrimp. Six isonitrogenous and isolipidic diets were formulated to supply 0, 1,000, 2,000, 4,000, 6,000, 10,000 mg/kg choline chloride, and dietary choline levels were analysed to be 1,317 (basal diet), 1,721, 2,336, 3,294, 5,421 and 9,495 mg/kg, respectively. Dietary choline levels significantly influenced percent weight gain (PWG) and protein efficiency ratio (PER), with the highest PWG was observed in shrimp fed the 5,421 mg/kg choline diet. However, there were no significant differences in proximate composition of whole body and muscle. Shrimp fed the diet containing 2,336 mg/kg choline had lower HDL and LDL in haemolymph than those fed the basal diet (1,317 mg/kg diet). Dietary choline prevented the accumulation of free radicals and improved antioxidant capacity by increasing catalase activity and reducing malondialdehyde content. Based on broken‐line regression and quadratic regression analysis between PWG against dietary choline levels, the optimal choline requirements were estimated to be 3,254.1 and 6,488.3 mg/kg for juvenile L. vannamei, respectively.     

Assessment of the relative contribution of dietary nitrogen from fish meal and biofloc meal to the growth of Pacific white shrimp (Litopenaeus vannamei)

The relative contribution of the dietary nitrogen supplied by fish meal and a biofloc meal to the growth of Pacific white shrimp was evaluated using stable isotope analysis. Biofloculated material was obtained from an experimental tilapia culture system. Five formulated diets were supplied. Two of them consisted in isotopic controls having only fish meal or biofloc meal as protein source. Three mixed diets were formulated with varying proportions of these ingredients on a dietary nitrogen basis (75:25, 50:50 and 25:75). At the end of the trial, survival rates were similar (92–100%) but significant differences in mean final weight were observed and a negative correlation between the inclusion of biofloc meal and weight gain was evidenced. Mean final weight in shrimp fed on diet containing only fish meal was 2.8 g, while mean final weight of animals fed on diet containing 50% biofloc was 1.9 g. Isotopic mixing models indicated that all diets contributed higher proportions of dietary nitrogen from fish meal than from biofloc meal. Dietary nitrogen available in diets containing 25%, 50% and 75% of biofloc meal was incorporated in muscle tissue as 5%, 41% and 64% respectively. Diet supplying 25% of nitrogen from biofloc was the only mixed diet eliciting growth comparable to diet containing only fish meal. Lower growth and nitrogen deposition in shrimp fed on diets containing high proportions of biofloc meal were possibly associated to the use of only two protein sources and a restriction of essential amino acids.     

Effect of dietary lipid level on growth,lipid metabolism and health status of the Pacific white shrimp Litopenaeus vannamei at two salinities

Three isonitrogenous diets containing 60 g kg^–1, 90 g kg^–1 or 120 g kg^–1 lipid were formulated and fed to the Litopenaeus vannamei (2.00 ± 0.08 g) under two salinities (25 or 3 psu) in triplicate for 8 weeks. Shrimp fed 90 g kg^–1 lipid had higher weight gain and specific growth rate than shrimp fed the other two diets regardless of salinity, and the hepatosomatic index increased with increasing dietary lipid at both salinities. The shrimp at 3 psu had significantly lower survival and ash content, higher condition factor, weight gain and specific growth rate than the shrimp at 25 psu. Increasing dietary lipid level induced the accumulation of serum MDA regardless of salinity, and at 3 psu, it reduced the serum GOT and GPT activities and the mRNA expression of TNF‐α in intestine and gill of L. vannamei. The hepatopancreatic triacylglycerol lipase (TGL) and CPT‐1 mRNA expression showed the highest value in shrimp fed 90 g kg^–1 lipid diet at 3 psu. This study indicates that 120 g kg^–1 dietary lipid may negatively affect the growth and induce oxidative damage in shrimp, but can improve immune defence at low salinity; 60 g kg^–1 dietary lipid cannot afford the growth and either has no positive impact on the immunology for L. vannamei at 3 psu.