Cholesterol requirement and phytosterols efficiency in semi‐purified diets of juvenile Pacific white shrimp Litopenaeus vannamei

Studies pertaining to cholesterol requirement are limited based on the levels evaluated and statistical analysis. Two trials were conducted to refine cholesterol requirement in Pacific white shrimp Litopenaeus vannamei feeds. The basal diet was supplemented with graded levels of cholesterol (ranging from 0.48 to 1.85 g/kg) in trial 1. Since the shrimp (initial weight: 0.24 g) growth response was very linear without reaching a plateau, it was not clear whether the requirement was met. Hence, a second trial was designed using eight diets designed with an increased range of cholesterol levels from 0.45 to 4.57 g/kg of diets. The shrimp (0.38 g initial weight) reached a final weight of (4.31–7.43 g) or per cent weight gain from 1,014%–1,874% (n = 4) after 6 weeks. Saturation kinetic model and broken‐line models with linear or quadratic ascending portions were used to evaluate dose–response relationships of feed efficiency (FE), thermal‐unit growth coefficient (TGC), whole‐body cholesterol (CHOL) and cholesterol deposition (CHOLD) against dietary cholesterol. The cholesterol requirement of shrimp was estimated at 1.9 (1.1–4.3) g/kg, 1.7 (1.1–3.4) g/kg, 2.5 (2.3–2.9) g/kg and 2.7 (2.4–3.1) g/kg of shrimp diet for FE, TGC, CHOL and CHOLD, respectively. Additionally, based on improved growth, the inclusion of phytosterol can serve as a partial replacement for cholesterol.     

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.     

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.     

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.     

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.     

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.     

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.     

Artificial substrates in zero‐water‐exchange culture system regulate the rearing performance of Pacific white shrimp Litopenaeus vannamei (Boone, 1931) under the winter indoor condition

Effects of artificial substrates in zero‐water‐exchange culture system on the rearing performance of Litopenaeus vannamei under winter indoor condition were investigated in this study. Growth, survival, feed conversion rate (FCR), production rate of L. vannamei and water quality were compared between artificial substrate‐treated group (AST) and control group (without artificial substrates presented in the rearing environment). Artificial substrates can significantly improve the water quality, the ammonia and nitrite‐N concentrations in the AST group were significantly lower than in the control group (P < 0.05), and the total heterotrophic bacteria and Vibrio spp. were also significantly lower in the AST group (P < 0.05). The survival, growth and production rate of L. vannamei in the AST group were significantly higher than in the control group (P < 0.05). Significantly lower FCR was observed in the AST group (P < 0.05). Results from this study indicate that the utilization of artificial substrates in the indoor shrimp culture system could effectively control the water quality, improve the survival and growth of shrimp and significantly reduce the FCR. This study provides a guideline for employing artificial substrates in rearing of shrimp in the zero‐water‐exchange culture system under lower temperature, which could be applicable to other similar species.     

Virulence and genotypes of white spot syndrome virus infecting Pacific white shrimp Litopenaeus vannamei in north‐western Mexico

White spot syndrome virus (WSSV) has caused substantial global economic impact on aquaculture, and it has been determined that strains can vary in virulence. In this study, the effect of viral load was evaluated by infecting Litopenaeus vannamei with 10‐fold serial dilution of tissue infected with strain WSSV Mx‐H, and the virulence of four WSSV strains from north‐western Mexico was assessed along with their variable number of tandem repeat (VNTR) genotypes in ORF75, ORF94 and ORF125. The LD₅₀ of the Mx‐H strain was a dilution dose of 10^?7.5; the mortality titre was 10^9.2 LD₅₀ per gram. In shrimp injected with 10^2.5 to 10^6.5 LD₅₀, no significant virulence differences were evident. Using mortality data, the four WSSV strains grouped into three virulence levels. The Mx‐F strain (intermediate virulence) and the Mx‐C strain (high virulence) showed more genetic differences than those observed between the Mx‐G (low‐virulence) and Mx‐H (high‐virulence) strains, in ORF94 and ORF125. The application of high‐viral‐load inocula proved useful in determining the different virulence phenotypes of the WSSV strains from the Eastern Pacific.     

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.     

Pond production of Pacific white shrimp,Litopenaeus vannamei,fed grain distillers dried yeast

The objective of this study was to demonstrate the feasibility of four diets formulated to contain increasing levels (0, 50, 100 and 150 g kg^?1 of diet) of grain distillers dried yeast (GDDY) in production diets for Litopenaeus vannamei, reared in outdoor tanks or production ponds. The production pond trial was carried out in 16, 0.1‐ha ponds using four replicates per diet. Juvenile shrimp (38.1 ± 4.26 mg, initial weight) were stocked at 30 shrimp m^?2 for a 16‐week period. The same four diets and a commercial reference diet were offered to shrimp maintained in outdoor tanks over a 12‐week period. A total of 20 tanks were stocked with juvenile shrimp (3.05 ± 0.22 g, initial weight) obtained from production ponds at a density of 30 shrimp per tank (40 shrimp m^?2). At the conclusion of these trials, mean final weight ranged from 19.77 to 23.05 g, yield ranged between 4760 and 5606 kg ha^?1, survival ranged from 69.6% to 89.4%, and feed conversion ratio (FCR) was between 1.02 and 1.23. Shrimp reared in the outdoor tanks confirmed the results of the pond trial. Mean final weight ranged between 18.12 and 18.97 g, survival ranged from 93.3% to 98.3%, and FCR was between 1.25 and 1.29. In both trials, there were no significant differences regarding mean final weight, FCR and survival among dietary treatments. Based on this study, GDDY up to 150 g kg^?1 of diet can be used in L. vannamei commercial feed formulation.     

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.     

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.     

Partial replacement of fish meal with peanut meal in practical diets for the Pacific white shrimp,Litopenaeus vannamei

In this study, we replaced fish meal with peanut meal (PM) in isonitrogenous and isolipidic diets for Pacific white shrimp at inclusion levels of 0, 70, 140, 210, 280 and 350 g kg^?1. The diets were hand‐fed to three independent groups of shrimp three times a day over a 6‐week period. Shrimp fed PM diets at a level of 280 g kg^?1 or higher had lower per cent weight gain compared with those fed the basal diet, whereas shrimp fed PM diets at 140 g kg^?1 or higher had a lower feed utilization and protein efficiency ratio compared with shrimp fed the basal diet. The feeding rate in shrimp fed PM diets at 350 g kg^?1 and the survival and protease activity in shrimp fed PM diets at 210 g kg^?1 or higher were lower than that in shrimp fed the basal diet. Diets containing 280 g kg^?1 or higher of PM caused an increase in the whole‐body moisture content of the shrimp, but decreased whole‐body protein and ash contents compared with the basal diet. Nutrient digestibility was lower or tended to be lower in shrimp fed a PM diet compared with those fed the basal diet. The activities of peroxidase and acid and alkaline phosphatases in plasma decreased with increasing levels of PM inclusion up to 210 g kg^?1. Superoxide dismutase activity decreased at dietary PM levels of 280 g kg^?1 or higher. Aflatoxin B₁ residue in the muscle was not affected by any of the treatments and remained low. The data suggest that up to 140 g kg^?1 of PM could be included in practical diets for Pacific white shrimp.     

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.     

Effects of various dietary levels of copper hydroxychloride on growth performance and tissue response in Pacific white shrimp Litopenaeus vannamei fed practical diets

Two trials were conducted to evaluate the performance of Pacific white shrimp Litopenaeus vannamei offered diets containing various copper (Cu) levels from Cu hydroxychloride (Cu₂(OH)₃Cl) containing 58.81% copper in the clear water recirculating system. In both trials, the basal diet (360 g kg^?1 protein, 80 g kg^?1 lipid) containing approximately 10 mg Cu kg^?1 was primarily comprised of fishmeal, soybean meal, corn protein concentrate and whole wheat. In trial 1, test diets were produced supplementing the basal diet with 5, 10, 20, 40 and 60 mg Cu kg^?1 from Cu hydroxychloride. Four replicate groups of 15 shrimp per tank (initial weight 0.28 g) were offered diets in slight excess over 8 weeks. In trial 2, the basal diet was supplemented with 30, 90, 150, 210 and 270 mg Cu kg^?1 from Cu hydroxychloride. Seven replicate groups of 15 shrimp per tank (initial weight 0.22 g) were offered feed in slight excess over 7 weeks. At the end of the two growth trials, no significant differences were observed in final biomass, final mean weight, percentage weight gain, feed conversion ratio (FCR) and survival. In trial 1, the Cu concentrations of the carapace, hepatopancreas and whole shrimp linearly increased with increasing dietary Cu supplements. In trial 2, polynomial regression of Cu concentrations of the carapace, hepatopancreas and whole shrimp against analysed dietary Cu content indicated that a plateau was reached at 215 mg analysed Cu kg^?1. Results of this study indicate that there was no negative effect of high levels of Cu supplement with regard to growth and survival. Tissue levels generally increased up to around 200 mg Cu kg^?1 diet and then decreased, possibly indicating a shift in physiology.     

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.     

Evaluation of flash dried yeast as a nutritional supplement in plant‐based practical diets for Pacific white shrimp Litopenaeus vannamei

Growth and digestibility trials were conducted to evaluate the use of flash dried yeast (FDY) as a supplement in practical shrimp feeds. The six‐week growth trial evaluated the effects of FDY supplementation (0, 10, 20, 40 and 60 g kg^–1) in the plant‐based practical diet on the performances and proximate compositions of juvenile Pacific white shrimp, Litopenaeus vannamei. Final biomass, final mean weight, per cent weight gain (WG), feed conversion (FCR) and protein retention efficiency (PRE) of Pacific white shrimp were not significantly influenced when FDY was added up to 40 g kg^–1 of the diet. However, 60 g kg^–1 FDY supplementation significantly reduced growth, feed utilization and protein retention. Supplementation of FDY in the practical diets of Pacific white shrimp did not affect protein, moisture, lipid, crude fibre, and ash content of whole shrimp body. Apparent digestibility coefficients of energy, protein, and amino acid (AA) for FDY were determined using chromic oxide as an inert marker and the 70:30 replacement technique. The energy and protein digestibility of FDY were 38.20% and 53.47%, respectively, which were significantly lower than fishmeal (FM) and soybean meal (SBM) that were run at the same time. Most amino acid digestibility of FDY was lowest among the three ingredients tested. Results of this work indicated that energy, protein and amino acid digestibility of FDY are lower compared to SBM and FM, and high levels supplementation (≥60 g kg^–1) of FDY can cause negative effects to the growth response of Pacific white shrimp. Based on these results, further research regarding the effects of the low level inclusion of FDY in practical diets on immune responses of Pacific white shrimp is warranted.     

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.