A Comparison between dietary effects of Cuminum cyminum essential oil and Cuminum cyminum essential oil,loaded with iron nanoparticles,on growth performance,immunity and antioxidant indicators of white leg shrimp (Litopenaeus vannamei)

This study evaluates the effect of dietary cumin essential oil's (CEO) and cumin essential oil's loaded iron nanoparticles (CEO‐loaded NPs) on growth performance, haematological parameters and antioxidant status of white leg shrimp. Shrimps with the initial weight of 3.38 ± 0.014 g were allocated into 15 fibreglass tanks (300 L, 60 shrimps per tank) and were fed with five experimental diets containing 0 g/kg CEO (the control diet), 5 g/kg CEO, 10 g/kg CEO, 5 g/kg CEO‐NPs and 10 g/kg CEO‐NPs for 56 days. After 56 days of culture, the shrimps fed with CEO and CEO‐loaded NPs supplemented diets showed a more significant increase in growth performance (p < .05) than in the control diet. Total haemocyte count (THC) value, large‐granule cells (LGC), hyaline cells (HC)%, superoxide dismutase (SOD) and Alkaline phosphatase (AKP) activity were significantly higher (p < .05) in the shrimps fed with CEO and CEO‐loaded NPs supplemented diets than the shrimps fed with the control diet. Glutathione peroxidase (GPX) enzyme activities along with the glucose and cholesterol concentrations significantly decreased in the shrimps fed with the CEO and CEO‐loaded NPs supplemented diets more than those fed with the control diet. The results suggest that the dietary CEO‐loaded NPs can improve the growth performance, the health status and the antioxidant capacity of L. vannamei.     

Effects of different levels of dietary protein on the growth performance,nutrient utilization,digestive enzymes and physiological status of white shrimp,Litopenaeus vannamei juveniles reared in inland saline water

A 60‐day feeding trial was conducted to illustrate the effect of dietary protein levels on the growth and physio‐metabolic responses of juvenile Litopenaeus vannamei reared in inland saline water (ISW). Six isoenergetic (15 MJ/kg) and isolipidic (60 g/kg) diets with graded level of crude protein viz., 240 (T240), 260 (T260), 280 (T280), 300 (T300), 320 (T320) and 340 (T340) g/kg diet were formulated. Significantly higher (p < .05) weight gain (%), specific growth rate, with lower food conversion ratio were found in T320 and T340 groups. The protein utilizing efficiency and whole‐body protein content were significantly higher (p < .05) in the T320 group. Trypsin activity increased with the increasing dietary CP level but amylase activity decreased with the increasing dietary CP level. Transaminase enzymes, haemolymph protein and haemocyanin were elevated in T320 and T340 groups. The lactate dehydrogenase and malate dehydrogenase activities were significantly higher (p < .05) in the T240 group. Shrimp of T240 group had the lowest (p < .05) glycogen and total haemocyte count with highest (p < .05) haemolymph glucose and antioxidant enzymes activities than the other groups. Based on the results, feeding 320 g CP/kg is found to be optimum for supporting maximum growth and health status of L. vannamei reared in ISW at 8 g/L salinity. The finding of the present study will help in developing a low‐cost feed for L. vannamei reared in ISW.     

Gut bacterial community profile in Pacific white shrimp Litopenaeus vannamei following 5‐aminolevulinic acid supplementation

Changes in the diet have been previously reported to alter the gut bacterial profile in several organisms, including shrimp. These shifts in microbial structure either promote beneficial effects to the host or cause diseases. Supplementation of 5‐aminolevulinic acid (5‐ALA), the precursor of tetrapyrroles, has been previously reported to enhance shrimp's immune response. To know whether 5‐ALA has effects on the gut bacterial structure in Pacific white shrimp (Litopenaeus vannamei), we investigated the bacterial communities in the intestine and stomach of L. vannamei using high‐throughput Illumina sequencing of 16S rRNA gene libraries. One week of 5‐ALA supplementation altered the bacterial community structure (beta diversity) in both tissues, as shown by the results of multi‐dimensional scaling (MDS) plots and analysis of similarities (ANOSIM). DESeq2 analysis revealed enrichment of differentially abundant taxa in the 5‐ALA group (e.g. Enhydrobacter and Oceaniovalibus) and control group (e.g. Tenacibaculum and Mycobacterium). Metagenomic predictions suggested that the control group had more KEGG pathways associated with ‘metabolism’ than the 5‐ALA group. This study suggests that 5‐ALA supplementation potentially promotes the formation of a beneficial bacterial community structure in shrimp. This is the first report on the effect of 5‐ALA supplementation on the bacterial community profile in any organism.     

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.     

Effect of polysaccharides extracts of algae Ulva rigida on growth,antioxidant, immune response and resistance of shrimp,Litopenaeus vannamei against Photobacterium damselae

This study was carried out to investigate the effect of water‐soluble polysaccharides extract of algae Ulva rigida (WPU) as dietary supplement on growth performance, antioxidant enzyme activity, lysozyme and phenoloxidase activity, and resistance of shrimp (Litopenaeus vannamei) subjected to bacterial infection with Photobacterium damselae. Three replicate groups of shrimp (1.0 g) were fed four diets containing four levels, 0 or control, 0.5, 1 and 1.5 g/kg of WPU for 8 weeks over the growth trial. Thereafter, 30 shrimps from each dietary treatment were infected with bacteria P. damselae to evaluate disease resistance of infected shrimp. The results of this study showed that WPU was effective as a growth promoter for L. vannamei. The best growth rate was observed in shrimp fed 1.5 g/kg of WPU diet. Regarding antioxidant defences, the diets supplemented with three levels of WPU stimulated glutathione peroxidase and catalase activates in experimental shrimps. MDA content of L. vannamei‐fed diet containing WPU 1.5 and WPU 1.0 was lower than WPU 0 and WPU 0. 5 diets. Also, lysozyme and phenoloxidase activities of shrimp receiving WPU at 1.0 and 1.5 level were significantly higher than those fed WPU 0 and WPU 0.5 diets. In addition, using WPU extract in all diets decreased mortality in L. vannamei in a dose‐dependent manner after challenge with P. damselae. These results suggest that incorporation of water‐soluble polysaccharides from green algae U. rigida at 1.5 g/kg doses improves growth and antioxidant activity and enhances the immune responses in shrimp L. vannamei.     

Oral Administration of Hot‐water Extract of Tropical Brown Seaweed,Sargassum cristaefolium,to Enhance Immune Response,Stress Tolerance,and Resistance of White Shrimp,Litopenaeus vannamei,to Vibrio parahaemolyticus

The efficacy of hot‐water extract of tropical brown seaweed, Sargassum cristaefolium (SCE), supplemented in diets on immune response, stress tolerance, and disease resistance of Litopenaeus vannamei to Vibrio parahaemolyticus was evaluated. Shrimp were fed diets containing graded levels of SCE (0, 250, 500, 750, and 1000 mg/kg). The results showed that shrimp fed all diets containing SCE had significantly higher (P < 0.05) immune response in total hemocyte count (THC), differential hemocyte count (granular and hyaline cells), and phagocytic activity than those of shrimp fed the control diet. Similarly, in low dissolved oxygen stress tolerance test and the challenge test with V. parahaemolyticus, survival rates of shrimp fed all diets containing SCE were significantly higher (P < 0.05) (83–93% in stress test and 27–47% in challenge test) than those of shrimp fed the control diet (77 and 3.3%, respectively). These results suggest that oral administration of SCE at 500 and 750 mg/kg can be effectively used to enhance immune response, stress tolerance, and resistance of white shrimp, L. vannamei, against V. parahaemolyticus infection. These findings also confirm that using dietary SCE as immunostimulant is effective at increasing the nonspecific immune system in penaeid shrimp, 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.     

Assessment of dietary lipid sources in tropical gar,Atractosteus tropicus larvae: Growth parameters and intermediary lipogenic gene expression

Five experimental diets containing different lipid sources, fish oil (D1), soybean lecithin (D2), corn oil (D3), canola oil (D4) and olive oil (D5), were evaluated in Atractosteus tropicus larvae for the relative gene expression of the enzymes fatty acid synthase (fas), acetyl‐CoA carboxylase 1 (acc1) and carnitine palmitoyltransferase 1C (cpt1c), in addition to their effects on larval growth, survival and cannibalism during a 30‐day feeding trial. Higher growth and survival were obtained in treatments D1 and D2, and lower performance in diets D3, D4 and D5. The highest levels of expression of fas and acc1 occurred in larvae fed with D1, which contained high amounts of n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFA), mainly DHA and EPA FA are regulators of lipogenesis. The higher cpt1c expression in plant‐based diets is attributed to the fact that these diets are rich in α‐linolenic acid (ALA) and low DHA, EPA and ARA levels that favour ß‐oxidation. In conclusion, the diets with fish oil (D1) and soybean lecithin (D2) were the best treatments for larval growth, survival and cannibalism and thus appear to meet both lipid and energy requirements of A. tropicus larvae, meanwhile the use of vegetable oils influences the expression of intermediary lipogenic genes.     

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.     

Effects of dietary n‐3 PUFA supplements on composition of n‐3 PUFA and expression of fatty acid elongase 5 (AJELOVL5) in sea cucumber,Apostichopus japonicus

This is the first comprehensive study on the effect of dietary polyunsaturated fatty acid (PUFA) levels on the expression of fatty acid elongase 5 (AJELOVL5), PUFA composition, and growth in juvenile sea cucumbers. The specific growth rate (SGR_w) was improved in n‐3 PUFA‐rich diets compared to low n‐3 PUFA diets. AJELOVL5 expression was apparently upregulated in juveniles fed lower PUFA diets relative to higher PUFA diets, with higher expression in the body wall and respiratory tree of juveniles fed diets without ɑ‐linolenic acid (ALA, 18:3n‐3) compared to juveniles fed higher ALA level diets; similar results were also detected in juveniles fed diets with lower eicosapentaenoic acid (EPA, 20:5n‐3), docosahexaenoic acid (DHA, 22:6n‐3), and none of ALA, EPA, or DHA respectively. The concentrations of ALA, EPA, and DHA in tissues were positively related to the content of dietary corresponding PUFA, with higher ALA content in juveniles fed diet ALA_12.71 than in the ALA_7.46 and ALA₀ groups. Similar results were also obtained in sea cucumber fed diets enriched with either EPA or DHA. Interestingly, considerable levels of EPA and DHA were found in the tissues of juveniles fed diets of CK₀ and DHA₀, with no specific input of EPA or DHA, showing that the sea cucumber was capable of biosynthesizing EPA and DHA from their corresponding precursors as ALA and linoleic acid (LA, 18:2n‐6).     

Immune responses by dietary supplement with Astragalus polysaccharides in the Pacific white shrimp,Litopenaeus vannamei

Two experiments were conducted to investigate the immune‐enhancing effect of dietary supplement with Astragalus polysaccharides (APS) on the Pacific white shrimp, Litopenaeus vannamei. In experiment 1, the optimal APS dose was determined based on the immune responses of shrimps fed APS diet for 30 days. In experiment 2, the effect of APS supplementation on immune response of shrimp suffering white spot syndrome virus (WSSV) challenge was determined. Results showed that the total haemocyte count and phagocytic activity in shrimps fed APS diets significantly (p < .05) increased in comparison with those fed the basal diet. Dietary supplement with APS markedly (p < .05) increased the activity of phenoloxidase (PO), total superoxide dismutase (SOD), lysozyme (LZM), acid phosphatase and alkaline phosphatase in shrimp hemolymph, but decreased the maleic dialdehyde (MDA) content. Significantly higher (p < .05) activity on PO, SOD and LZM and lower (p < .05) MDA content have also been found in shrimps suffering WSSV challenge. Therefore, APS could be used as a safe and effective feed additive in shrimp aquaculture, and the optimal dose of APS for the Pacific white shrimp was suggested to be 0.2 g/kg based on our results.     

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.     

Effects of dietary n‐3HUFA on different growth stages of the white shrimp,Litopenaeus vannamei: Growth,haematological characteristics,enzyme activities and fatty acid profiles

This study investigated the effects of n‐3 high unsaturated fatty acid (n‐3HUFA) levels on the growth performance, antioxidant enzyme activities and fatty acid profiles of both subadult and adult Litopenaeus vannamei (L. vannamei). Seven iso‐nitrogenous and iso‐lipidic diets were used, containing n‐3HUFA concentrations of 1.6 (control), 4.8, 7.4, 13.9, 23.9, 29.2 and 34.4 g/kg, respectively. Two 8‐week feeding trials were conducted to determine the dietary n‐3HUFA requirements of L. vannamei with an initial body weight of 4.25 ± 0.00 g (subadults) and 8.50 ± 0.01 g (adults). The results showed that the dietary n‐3HUFA level significantly affected the weight gain rate (WGR), specific growth rate, the feed conversion ratio and the hepatosomatic index (HSI) (p < 0.05), but did not significantly affect the survival rate (p > 0.05). At appropriate level, dietary n‐3HUFA improved growth performance and HSI of both subadult and adult L. vannamei. Both subadults and adults showed significant differences in body composition (p < 0.05), except for moisture and crude ash (p > 0.05). Cholesterol and low‐density lipoprotein significantly decreased with increasing dietary n‐3HUFA both in subadults and adults (p < 0.05); however, triglyceride showed no significant change (p > 0.05). High‐density lipoprotein (HDL) in subadults was significantly affected by dietary n‐3HUFA (p < 0.05), but followed no apparent regularity; HDL significantly changed in adults and showed an upward trend followed by a downward trend (p < 0.05). There was no significant effect on aspartate transaminase (AST) activity in subadults, but AST in adults and alanine transaminase (ALT) in subadults and adults were significantly affected (p < 0.05). Dietary n‐3HUFA significantly affected serum polyphend oxidase, malic dehydrogenase, alkaline phosphatase, superoxide dismutase and sodium‐potassium adenosine triphosphatase enzyme activities in gills (p < 0.05). The fatty acid composition of the shrimp tissue was associated with the fatty acid composition of the diet. Dietary n‐3HUFA supplementation significantly improved the contents of tissue ∑HUFA and n‐3HUFA, increased the n‐3/n‐6 ratio in the tail muscle and decreased the contents of tissue polyunsaturated fatty acid and saturated fatty acid (p < 0.05). Based on the WGR, the broken‐line equations indicated that the optimum requirements of dietary n‐3HUFA were determined to be 9.0 and 5.1 g/kg for subadult and adult L. vannamei, respectively.     

Innate immune response and gene expression of Scylla paramamosain under Vibrio parahaemolyticus infection

An epidemic of ‘milky disease’ in the mud crab (Scylla paramamosain) generally breaks out in the fall when the crab is near maturity, resulting in large economic losses in crab farming. Vibrio parahaemolyticus has been proven to be one of the major pathogens. In this study, the mud crabs were challenged with V. parahaemolyticus, and their innate immune responses were investigated in terms of total haemocyte counts (THCs), haemocytic enzyme activities and gene expression levels during a 114‐h period. The THCs of the mud crabs decreased significantly after 42 h of exposure. The activities of the haemocytic enzymes, including acid phosphatase‐alkaline phosphatase, phenoloxidase, superoxide dismutase (SOD), peroxidase and nitric oxidase synthethase, were significantly enhanced during the challenge course. The gene expression levels also significantly increased for all tested genes (proPO, Cu/Zn‐SOD, Prx, LYS, CRU and ALF) with the exception of CAT down‐regulated expression. The results may imply that the immune responses of the mud crab could be activated by the pathogens, and the data here will provide many clues for further systematic investigation of ‘milky disease’ caused by V. parahaemolyticus and the disease prevention in mud crab S. paramamosain.     

Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone)

Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies.     

Effects of different carbon sources and dietary protein levels in a biofloc system on growth performance,immune response against white spot syndrome virus infection and cathepsin L gene expression of Litopenaeus vannamei

A 5‐week study was performed to evaluate the effect of spoilage date extract (SDE) as the biofloc carbon source on Litopenaeus vannamei (5.4 ± 0.3 g) performance. The two levels of dietary protein (15% and 25% crude protein) and two carbohydrate sources (molasses‐M and SDE‐P) were tested including: M15, M25, P15 and P25. The minimum (0.2 ± 0.0 mg/L) and the maximum (0.5 ± 0.0 mg/L) of total ammonia nitrogen were observed in the P15 and M25 groups respectively. The highest protein efficiency ratio (6.1 ± 0.3) and protein productive value (112.3 ± 5.8%) were found in the P15 group (p < 0.05). No significant difference was found between biofloc treatments in the expression of cathepsin L gene in hepatopancreas (p > 0.05). The number of total haemocyte count (THC), semigranular cells (SGC) and granular cells (GC) of shrimp in SDE‐based biofloc treatments was relatively higher than those in molasses‐based biofloc treatments. Following the white spot syndrome virus (WSSV) challenge, a significant decrease in THC, SGC, GC and hyaline cell values was observed in all treatments (p = 0.001). Plasma biochemical parameters were significantly influenced by dietary protein levels, biofloc carbon sources as well as WSSV challenge test. In conclusion, SDE successfully could be used as an alternative carbon source for establishing a biofloc system in L. vannamei production.     

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

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

Immune responses of whiteleg shrimp,Litopenaeus vannamei (Boone, 1931), to bacterially expressed dsRNA specific to VP28 gene of white spot syndrome virus

In this study, dsRNA specific to VP28 gene of white spot syndrome virus (WSSV) of shrimp was synthesized in Escherichia coli in large scale and studied the immune response of shrimp to dsRNA‐VP28. The haematological parameters such as clotting time and total haemocytes counts, and immunological parameters such as prophenoloxidase (proPO), superoxide dismutase (SOD), superoxide anion (SOA) and malondialdehyde content, as well as the mRNA expression of ten immune‐related genes were examined to estimate the effect of dsRNA‐VP28 on the innate immunity of Litopenaeus vannamei. The activities of proPO, SOA and SOD significantly increased in haemocyte after dsRNA‐VP28 treatment, whereas MDA content did not change significantly. Among the ten immune‐related genes examined, only the mRNA expression of proPO, cMnSOD, haemocyanin, crustin, BGBP, lipopolysaccharides (LPs), lectin and lysozyme in haemocytes, gill and hepatopancreas of L. vannamei, was significantly upregulated at 12 h after dsRNA‐VP28 treatment, while no significant expression changes were observed in Toll receptor and tumour receptor genes. The increase of proPO and SOD activities, and SOA level and mRNA expression level of proPO, cMnSOD, haemocyanin, crustin, BGBP, LPs, lectin and lysozyme after dsRNA‐VP28 stimulation indicate that these immune‐related genes were involved in dsRNA‐VP28‐induced innate immunity in shrimp.     

Growth performance,haematological parameters,antioxidant status and salinity stress tolerance of juvenile Pacific white shrimp (Litopenaeus vannamei) fed different levels of dietary myo‐inositol

A 10‐week growth trial was run to evaluate effects of myo‐inositol (MI) on growth performance, haematological parameters, antioxidative capacity and salinity stress tolerance of Litopenaeus vannamei. Six practical diets supplemented with graded levels of MI (designated as MI0, MI600, MI1200, MI2400, MI 3600 and MI4800 for 448.8, 974.2, 1568.0, 2810.6, 3835.5 and 4893.6 mg/kg diet, respectively) were fed to six replicate groups of L. vannamei (mean initial body weight 0.63 ± 0.00 g). The results showed that significant increment of growth performance was observed in shrimp fed MI600 diet than those fed MI1200 diet. Lipid concentration in whole body of the shrimp fed MI600 diet was significantly increased. Shrimp fed MI0 diet had lower total protein (TP) as compared to shrimp fed the MI‐supplemented diets (except MI4800 diet). In general, lower activities of antioxidant enzymes and higher malondialdehyde (MDA) content in haemolymph and hepatopancreas were recorded in shrimp fed MI0 diet, compared to those fed the MI‐supplemented diets. Reduced survival after 7‐h salinity stress was present in shrimp fed MI0 diet as compared to those fed MI4800 diet. Dietary MI requirement for glutathione peroxidase activity of L. vannamei was 2705 mg/kg diet.