Ingestion of food pellets containing Escherichia coli overexpressing the heat‐shock protein DnaK protects Penaeus vannamei (Boone) against Vibrio harveyi (Baumann) infection

Feeding aquatic animals with bacterial encapsulated heat‐shock proteins (Hsps) is potentially a new method to combat vibriosis, an important disease affecting aquatic animals used in aquaculture. Food pellets comprised of shrimp and containing Escherichia coli overexpressing either DnaK‐DnaJ‐GrpE, the prokaryotic equivalents of Hsp70‐Hsp40‐Hsp20, or only DnaK were fed to juveniles of the white leg shrimp Penaeus vannamei, and protection against pathogenic Vibrio harveyi was determined. Maintaining pellets at different temperatures for varying lengths of time reduced the number of live adhering E. coli, as did contact with sea water, demonstrating that storage and immersion adversely affected bacterial survival and attachment to pellets. Feeding P. vannamei with E. coli did not compromise their survival, indicating that the bacteria were not pathogenic to shrimp. Feeding P. vannamei with pellets containing bacteria overproducing DnaK (approximately 60 cells g^?1 pellets) boosted P. vannamei survival twofold against V. harveyi, suggesting that DnaK plays a role in Vibrio tolerance. Pellets containing DnaK were effective in providing protection to P. vannamei for up to 2 weeks before loss of viability and that DnaK encapsulated by these bacteria enhanced shrimp resistance against Vibrio infection.     

Fatty Acid Profiles of Different Shrimp Species: Effects of Depth of Catching

The aim of this study was to investigate fatty acid profiles of different shrimp species caught from deep water and shallow water. The shrimp species investigated in the study were Aristeus antennatus, Aristeomorpha foliacea, Plesionica martia, Parapenaeus longirostris, and Plesionica edwardsi from deep water; and Metapenaeus monoceros, Penaeus semisulcatus, Penaeus kerathurus, and Penaeus japonicus from shallow water.

Fatty acid composition of shrimps varied. The main fatty acids were C18:1n9, C16:0, C25:6n3, C22:5n3, and C18:0. Saturated, monounsaturated, and polyunsaturated fatty acid (PUFA) contents of P. longirostris, P. edwardsi, and M. monoceros were markedly different, respectively. The ratio of n6/n3 of the edible tissue of M. monoceros was found to be 0.795, whereas this value was 0.152 in A. foliacea. Among the species studied, the highest docosahexaenoic acid + eicosapentaenoic acid value was found for P. kerathurus. The levels of PUFAs of shallow water shrimps (ranging from 33.44 to 42.77%) were found to be higher than those of deep water shrimps (ranging from 29.68 to 33.95%). Marine animals in the upper water layers gain nutrition through phytoplankton which provides n-3 PUFA depending on solar energy. Shallow water shrimp species provide a satisfying amount of PUFA.     

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.     

A diet of fructose‐enriched Artemia improves the response of juvenile Litopenaeus vannamei shrimp to acute low‐salinity challenge

The Pacific white shrimp (Litopenaeus vannamei), as an economically important species, has been reared in low‐salinity water during the last decade. To investigate how juvenile L. vannamei shrimp fed with fructose‐enriched Artemia respond to acute low‐salinity stress, the shrimp were randomly divided into four treatment groups, three groups were fed with Artemia enriched with either 100, 200 and 300 mg L^?1 of fructose and a control group fed with Artemia with no enrichment for 10 days. The results showed that the 300 mg L^?1 fructose group demonstrated the maximum survival rate and glycogen content. Additionally, the 300 mg L^?1 fructose group showed significantly higher Na⁺/K⁺‐ATPase activity, total antioxidant capacity, and expression levels of Na⁺/K⁺‐ATPase α‐subunit, V‐H ATPase α‐subunit, hexokinase, phosphofructokinase, antioxidants (SOD, CAT, GPX) and Hsp70 mRNA when compared with the control group. Furthermore, after exposure to low salinity, the mRNA levels of phosphofructokinase, V‐H ATPase α‐subunit, GPX, p38, JNK and Rac1 stayed constant in shrimp fed with fructose‐enriched Artemia but changed significantly in the control group. Thus, a diet of fructose‐enriched Artemia can improve the osmoregulation and survival of juvenile L. vannamei shrimp exposed to low salinities.     

Slack regulation compliance in the Mediterranean fisheries: a paradigm from the Greek Aegean Sea demersal trawl fishery,modelling discard ogives

Annual discard ogives were estimated using generalised additive models (GAMs) for seven demersal fish species (or taxa) and deep‐water rose shrimp, Parapenaeus longirostris (Lucas). Analysis was based on data collected on board commercial bottom trawlers in the central Aegean Sea from 1995 to 2008. Length of specimens and fishing depth (along with year) were the variables that had the most profound effect on the proportion of fish discarded. Compliance with the established minimum catch size of marine organisms (MS) was very low, a fact attributed to the low selectivity of currently used mesh sizes, the market demand for undersized fish, as well as the low control and enforcement effort.     

Heat-shock protein70 gene expression in four hatchery Pacific Abalone Haliotis discus hannai Ino populations using for marker-assisted selection

Heat shock proteins (Hsps) are molecular chaperones that help organisms cope with stressful conditions. Here, we report on the growth rates and Hsp70 expressions in inbred and hybrid populations of abalone Haliotis discus hannai Ino. In abalone, inbred populations expressed more Hsp70 than hybrid populations at all temperatures, except at very high temperatures close to the physiological limit. At benign temperatures, there was a clear trend towards higher Hsp70 expression in inbred than hybrid populations, whereas at higher temperatures, a trend in the opposite direction was observed. The temperature of maximal Hsp70 expression (T_peak) varied with the population type. The T_peak of inbred populations (26°C) was lower than that of the hybrid populations (28°C). The maximal inducible Hsp70 of inbred populations was higher than that of hybrid populations. The results showed a trend towards higher expression in inbred population at a lower temperature. These results provide direct experimental evidence that hybrids can cope with the intrinsic stress even at non‐stressful temperatures. The constitutive Hsp70 may therefore be used for marker‐assisted selection in a breeding programme.     

Molecular characterization and expression analysis of heat shock proteins 40, 70 and 90 from kuruma shrimp <Emphasis Type="Italic">Marsupenaeus japonicus</Emphasis>

Heat shock proteins (HSPs) are proteins that are expressed more strongly when the cells are exposed to physiological and stressful conditions. In this study, the full-length cDNAs of heat shock proteins 40 (MjHSP40), 70 (MjHSP70) and 90 (MjHSP90) were cloned from kuruma shrimp Marsupenaeus japonicus. The open reading frames (ORFs) of the cDNA clones have lengths of 1,191, 1,959 and 2,172 bp and encode 396, 652 and 723 amino acid residues, respectively. The predicted MjHSP40 amino acid sequence contains a J domain, a glycine/phenylalanine-rich region, and a central domain containing four repeats of a CxxCxGxG motif, indicating that it is a type I HSP40 homolog. The signature sequences of the HSP70 and HSP90 gene families are conserved in the MjHSP70 and MjHSP90 amino acid sequences. The deduced amino acid sequences of MjHSP70 and MjHSP90 share high identity with previously reported shrimp HSP70s and HSP90s, respectively. The expression of MjHSP90 mRNA increased at 32°C. Additionally, the expressions of MjHSP40, MjHSP70 and MjHSP90 mRNAs increased in defense-related tissues (i.e., hemocytes and lymphoid organ) when the shrimp were challenged with white spot syndrome virus.     

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.     

Effect of acclimation temperature on thermoregulatory behaviour,thermal tolerance and respiratory metabolism of Lutjanus guttatus and the response of heat shock protein 70 (Hsp70) and lactate dehydrogenase (Ldh‐a) genes

This study evaluated the effect of different environmental temperatures in the physiology of Lutjanus guttatus juveniles by analysing their thermoregulatory behaviour, thermal tolerance, oxygen consumption rates and thermal metabolic scope. Jointly, the effect of acclimation and critical temperatures on heat shock protein 70 (Hsp70) and lactate dehydrogenase (Ldh‐a) gene expressions were also analysed using acclimation temperatures of 20, 23, 26, 29 and 32°C. The results showed that the final preferred temperature in juvenile snapper was 26°C with a thermal window of 336.5°C², which was related to an optimal temperature for their physiology determined by the routine metabolic rate and thermal metabolic scope. At temperatures from 20 to 26°C, the routine metabolic rate and Hsp70 and Ldh‐a genes had the lowest values related to a basal expression level. At acclimation temperatures from 29 to 32°C and after critical thermal maximum (CT_max) limit, the relative expression of Hsp70 and Ldh‐a genes increased significantly, but the main response at CT_max was the upregulation of Hsp70 gene.     

Functional Properties of White Shrimp (Litopenaeus vannamei) By-Products Protein Recovered by Isoelectric Solubilization/Precipitation

The processing of fish in general entails quantities of by-products, such as exoskeleton and cephalothorax of shrimp, which may reach 70%. Protein recovered from by-products has been investigated in recent years for its ability to offer more nutritious foods and improved functional properties. The objective of the present study was the use of the pH shifting process to recover proteins from solubilization and precipitation that are present in the by-products (heads and shells) of white shrimp (Litopenaeus vannamei). The proximal composition of the by-products presented 58.5% protein (d.b). The isoelectric point used to precipitate the protein was defined at pH 4. The protein concentrate presented 73.1% of protein. The yield of the protein concentrate mass resulted in 47.8% (w.b). The greatest solubility found for the protein concentrate was 51.3% at pH 8. The oil and water retention capacity resulted in values of 8.5 and 2.5 mL/g of protein, respectively, and the highest emulsifying capacity was found at pH 8. The by-products and concentrate amino acids profiles confirmed the presence of 16 amino acids. In general, the shrimp protein concentrate obtained showed the potential to be included as an ingredient in food formulations.     

Effects of temperature and dietary protein on gene expression of Hsp70 and Wap65 and immunity of juvenile mirror carp (Cyprinus carpio)

Juvenile mirror carp were fed diets containing 303.4, 321.7, 341.2, 361.0 and 379.1 g kg^?1 proteins, respectively, and reared at different water temperatures (18, 23 and 28°C) for 60 days. Gene expression of heat shock protein gene (Hsp70) and the warm temperature acclimation‐related 65 kDa protein gene (Wap65), immunity and antioxidant status in the carp were investigated. Results indicated that the contents of serum complement 3 (C3), complement 4 (C4) and immunoglobulin M (IgM), as well as activities of liver superoxide dismutase (SOD) and lysozyme (LSZ) were significantly enhanced with increasing dietary protein (P < 0.05), while content of malondiadehyde (MDA) decreased. Gene expression level of Wap65 in the liver significantly increased with dietary protein, while gene expression of Hsp70 decreased. The contents of C3, C4 and IgM, the activities of SOD and LSZ and gene expression level of Wap65 in the liver significantly increased with temperature. These results suggest that: Serum immune parameter, antioxidant enzymes and Hsp70 and Wap65 expression interact in fish to improve ability to adapt to the environment; and the optimal conditions for the immunity of carp are 348.1?354.5 g kg^?1 protein at 18°C, 352.3?364.9 g kg^?1 at 23°C and 360.2?364.3 g kg^?1 at 28°C, and the optimum temperature for carp is 23°C.     

Use of Microbial Flocs Generated from Tilapia Effluent as a Nutritional Supplement for Shrimp, Litopenaeus vannamei, in Recirculating Aquaculture Systems

Recovering nutrients in a fish effluent to be used as a supplemental feed for shrimp culture could ease constraints (e.g., environmental issues and high production cost) that have limited the US shrimp farming industry in the past. In this study under laboratory scale conditions, fish effluent was collected from a commercial tilapia farm and nutrients from the waste stream were offered as supplemental feed as either (1) untreated solids from tilapia effluent or (2) microbial flocs generated from the biological treatment of the effluent by reducing soluble chemical oxygen demand >80%. The first feeding trial demonstrated that microbial flocs contributed significantly (P < 0.05) to overall growth while untreated solids did not. Moreover, microbial flocs were larger and contained higher levels (P < 0.05) of protein. The second feeding trial investigated different feeding rates of commercial diets with and without microbial floc supplementation. Weekly measurements of mass and specific growth rates demonstrated that microbial flocs significantly (P < 0.05) contributed to shrimp performance. Weekly food conversion ratios were also reported. Water quality in shrimp systems during both studies was within safe levels, and no differences (P > 0.05) between treatments were observed for dissolved oxygen, nitrate‐nitrogen, nitrite‐nitrogen, pH, salinity, total ammonia nitrogen, and temperature.     

低氧胁迫和恢复对长吻鮠脑组织低氧应答基因、生理生化指标和食欲的影响

为了解长吻鮠脑组织应对低氧胁迫的调控机制，实验运用酶活性测定、H.E染色、qRT-PCR和TUNEL检测等方法，分析比较了低氧胁迫[(0.8±0.1) mg/L] 0、2、4、6 h(标示为H0、H2、H4和H6)和恢复[(7.3±0.5) mg/L]2、4、6 h (标示为R2、R4和R6)下长吻鮠脑组织低氧应答基因、生理生化指标和食欲基因的变化。结果显示，在低氧胁迫和恢复下，长吻鮠脑组织氧传感蛋白相关基因(HIFs、PHDs和Vhl)表达量整体呈现出先上升后下降趋势；呼吸代谢酶(HK、PK和LDH)活性在H0时显著升高，SDH和MDH活性在H6时显著降低，恢复溶解氧后，代谢模式由无氧呼吸逐渐转变为有氧呼吸；抗氧化酶(GSH-Px、CAT和SOD)和应激指标(MDA和LPO)在低氧2 h后逐渐升高，恢复溶解氧后氧化应激现象仍然存在。观察脑组织形态发现，在低氧胁迫下长吻鮠脑组织出现了神经细胞肿胀、空泡等受损现象，恢复溶解氧6 h后脑组织受损并未得到有效改善。随着低氧时间延长，脑组织细胞凋亡程度不断增加，凋亡相关基因(Bax、Caspase-3和p53)表达量显著升高，而Bcl-2基因表...     

Dietary fulvic acid effects on survival and expression of immune‐related genes in Litopenaeus vannamei challenged with Vibrio parahaemolyticus

The effects of fulvic acid (FA) on survival and immune‐related gene expression were investigated in Litopenaeus vannamei challenged with Vibrio parahaemolyticus by immersion. Shrimp were fed with different dietary FA concentrations (1, 2, 4 and 6 g/kg feed) for 20 days (first bioassay) or 8 days (second bioassay, 2 g/kg feed of FA added every 2 days) and then challenged with V. parahaemolyticus. In a third bioassay, the expression of three immune‐related genes (translationally controlled tumour protein [TCTP], superoxide dismutase [SOD] and heat‐shock protein 70 [HSP70]) in haemocytes or hepatopancreas of experimental shrimp was measured by real‐time quantitative PCR at 0, 6, 12, 24, 48, 72 and 96 hr after FA (2 g/kg feed) administration. Fulvic acid increased survival at a concentration of 2 g/kg feed supplied every two days. Interestingly, TCTP gene expression was upregulated, whereas gene expression of SOD and HSP70 was downregulated. In conclusion, dietary fulvic acid improves survival in white shrimp challenged with V. parahaemolyticus and modulates the immune response. Therefore, FA merits further evaluation as prophylactic treatment in commercial shrimp farms.     

Advances in the study of activity additivity of supplemented proteases to improve digestion of feed protein by Penaeus vannamei

In vitro assays used porcine or bovine trypsin as models of exogenous enzymes to determine functioning in the presence of enzymatic extracts from the digestive gland of whiteleg shrimp Penaeus vannamei. Using electrophoresis and zymograms, when enzymes from the shrimp were mixed in the absence of protein substrate, they hydrolysed the trypsin from bovine or porcine origin. Porcine or bovine trypsin, when mixed with shrimp enzymes in pH‐stat assays in the presence of shrimp commercial feed, fish meal, or casein, there was added activity to hydrolyse the protein substrate. Hydrolysis of protein substrate was twofold to threefold stronger if exogenous enzymes were added. Results suggest that porcine or bovine trypsin could be used as feed supplements for whiteleg shrimp P. vannamei to enhance hydrolysis of proteins in feeds, because the commercial enzymes contributed to the hydrolysis of the protein in the three substrates in the presence of shrimp enzymes.     

Effects of High Pressure on Myofibrillar Protein and Moisture Distribution of Shrimp (Solenocera melantho) Muscle

ABSTRACT

The effects of high pressure (100–500 MPa) treatment on myofibrillar protein (MP) contents and distribution of water in Solenocera melantho muscle were investigated. The results showed that the shrimp treated with 200 MPa had the highest shelling score, and the solubility of MP in meat of shrimp treated with 300 MPa was significantly reduced. Moreover, the surface hydrophobicity of MP increased with increasing pressure. Treatment of shrimp with 200–300 MPa high pressure partially converted the structure of MP from an α-helix structure to a β-sheet structure, allowing for greater exposure of the tryptophan residues present. Treatment of shrimp with 100–200 MPa led to a significant loss in the amount of immobilized water in shrimp muscle. However, the treatment did not significantly change the moisture distribution. Overall, pressures of 100 to 200 MPa are considered the most suitable for the processing of S. melantho.     

Effects of Major Water Quality Variables on Shrimp Production in Inland,Low‐Salinity Ponds in Alabama

Common water quality variables in nine, inland low‐salinity shrimp ponds in Alabama exhibited wide variation in concentrations among ponds and over time. Shrimp performance also varied considerably among ponds in 2008 as follows: survival, 16–128%; production, 928–5950 kg/ha; feed conversion ratio (FCR), 1.18–2.89. Measured water quality variables were not at concentrations high enough to be lethal to shrimp; but water temperature, dissolved oxygen, carbon dioxide, total ammonia nitrogen, calcium, and magnesium were occasionally outside optimum ranges for shrimp production and may have stressed shrimp. Survival and production both were positively correlated (P < 0.05) with increasing concentrations of methyl orange alkalinity, total alkalinity, and calcium hardness. Negative correlations (P < 0.05) between production and higher pH and water temperature may have resulted from lower water temperature and pH during final days of the crop in ponds harvested in October rather than from an actual effect of temperature and pH on growth. Nevertheless, those variables that were outside optimal ranges or correlated with shrimp survival or growth should be further investigated to ascertain whether or not excursions outside optimum ranges are harmful and to determine if observed correlations are causal.     

Investigation of geographic origin,salinity and feed on stable isotope profile of Pacific white shrimp (Litopenaeus vannamei)

Increasing numbers of mislabelled seafood products in the globalized market underline the need for approaches to identify the origin of these products. Pacific white shrimp cultured in different salinities (Litopenaeus vannamei) from 16 locations in China and USA were collected and differentiated based on δ¹³C and δ¹⁵N values. The δ¹³C and δ¹⁵N values of 16 commercial feeds used in shrimp culture were measured and effects of feed on C and N stable isotope composition in shrimp were investigated. Carbon and nitrogen isotopic analysis of the shrimp did not provide means to discriminate farmed shrimp from China and USA. However, the shrimp cultured in water with high salinity were enriched in ¹³C compared to shrimp cultured in freshwater and this provided another basis for discrimination. The correlation between δ¹³C in feed and shrimp was not significant, while the δ¹⁵N in feed and shrimp showed a trend of positive correlation.     

Effect of using sodium bicarbonate to adjust the pH to different levels on water quality,the growth and the immune response of shrimp Litopenaeus vannamei reared in zero‐water exchange biofloc‐based culture tanks

The bioflocs technology proved to be a sustainable technique used in zero‐water exchange shrimp culture systems. However, the pH and alkalinity may decrease due to the biofloc formation process and Nitrification. A 48‐day experiment was performed to investigate the effects of different pH (7.1–7.6; 7.6–8.1) conditions on water quality, the growth and the health status of shrimp in biofloc technology (BFT) through using sodium bicarbonate to adjust pH respectively. Two pH treatments and one control were compared: T0 — control, T1 — pH 7.6 — NaHCO₃, T2 — pH 8.1 — NaHCO₃, each treatment consisted of three replicate tanks (90 L water volume) and each replicate stocked with 30 shrimp (equivalent to 333 shrimp m^?3). Significant physical, chemical and biological differences (P < 0.05) were detected among treatments. At the end of the experiment, water quality, the growth and the immune response of shrimp in control were significantly lower (P < 0.05) than the other treatments. Moreover, the T2 treatment had a better performance in these three aspects. The results indicated that it was necessary to adjust the pH and alkalinity in the BFT, and a higher pH as well as alkalinity for shrimp growth and the stability of the BFT were more favourable.