Interactive effects of dietary astaxanthin and cholesterol on the growth,pigmentation, fatty acid analysis,immune response and stress resistance of kuruma shrimp (Marsupenaeus japonicus)

A 56‐day feeding trial was carried out to evaluate the interactive effects of chemically synthesized astaxanthin (Ax) and cholesterol (CHO) on the performances of kuruma shrimp juveniles. Six experimental diets containing 2 levels of Ax (0 and 0.6 g/kg diet) and 3 levels of CHO (0, 6 and 20 g/kg diet) were formulated in 2 × 3 factorial design. The results indicated that the interactions between dietary CHO and Ax were not significant on survival, growth performances, body proximate analysis, immune response and freshwater stress test. Significantly, better pigmentation performances and Ax contents of different parts of juveniles (whole body, muscle, carapace and head) were observed in higher Ax (Ax_0.6)‐supplemented groups. Meanwhile, interactions of dietary Ax and CHO were also found on colour reading parameters and whole‐body Ax contents. In addition, juveniles fed with the diet containing Ax at 0.6 g/kg diet in addition to CHO at 0, 6 and 20 g/kg diet showed better performances on digestive enzyme activities (protease, lipase and amylase) of hepatopancreas than 0 g Ax/kg diet supplemented with CHO at 0, 6 and 20 g/kg diet. It was concluded that dietary Ax and CHO functioned interactively.     

Effects of dietary astaxanthin and vitamin E and their interactions on the growth performance,pigmentation, digestive enzyme activity of kuruma shrimp (Marsupenaeus japonicus)

A 56‐day feeding trial was done to investigate the interactive effects of astaxanthin (Ax) and vitamin E (α‐Toc) on the performance of kuruma shrimp (Marsupenaeus japonicus). A 2 × 3 factorial experiment was conducted with six experimental diets containing two levels of Ax (0 and 0.6 g/kg diet [Ax₀ and Ax_0.6]) and three levels of α‐Toc (0, 0.2 and 1 g/kg diet [α‐Toc₀, α‐Toc_0.2 and α‐Toc₁]). The results indicated that growth performance was significantly (p < 0.05) increased in shrimp fed with the Ax_0.6 × α‐Toc_0.2 diet. Interactive effects between Ax and α‐Toc on the growth parameters were observed. Furthermore, pigmentation performance was significantly (p < 0.05) better in the Ax_0.6 groups. Interaction between Ax and α‐Toc was also found in the Ax content of shrimp body parts. Interestingly, dietary α‐Toc helped to reduce the Ax consumption rate, promote the absorption and increase the deposition of Ax in the muscle. Shrimps from the Ax_0.6 groups showed significantly (p < 0.05) improved hepatopancreatic digestive enzyme activities compared with those of the Ax₀ groups. Although no interactive (p > 0.05) effects were found between dietary α‐Toc and Ax on total haemocyte count and tolerance against freshwater, dietary Ax and α‐Toc supplementation showed better performance on these two parameters. It was concluded that dietary Ax and α‐Toc functioned synergistically, and the shrimp fed with the diet containing 0.6 g Ax/kg diet Ax and 0.2 g α‐Toc/kg diet showed improved growth and pigmentation performance compared with the other groups in the current study.     

Effects of dietary Nannochloropsis sp. powder and lipids on the growth performance and fatty acid composition of larval and postlarval kuruma shrimp,Marsupenaeus japonicus

The current study evaluated the effects of fish oil replacement with eicosapentaenoic acid‐enriched single‐cell microalgae in the diets of larval and postlarval kuruma shrimp. Experimental diets containing different level of Nannochloropsis sp. powder (10, 40, 70 g/kg) and lipids (2, 8, 10, 14, 35, 58 g/kg) were evaluated. The substitution of fish oil with algal powder significantly improved shrimp growth compared to that in the control group, with the highest final body weight recorded at 70 g/kg in larvae and 40 g/kg in postlarvae. Larvae fed algal lipids with a dry weight percentage in the diet of greater than 10 g/kg showed significantly lower performance than those in the control group. The neutral lipids in the tissue of the postlarvae that had been fed algal lipids had an improved fatty acid profile, as the content of highly unsaturated fatty acids increased compared to that in the other groups. The content of docosahexaenoic acid increased in the tissue, indicating the possible occurrence of the metabolism and accumulation of Nannochloropsis sp. essential fatty acids. Algal powder could replace FO up to 140 g/kg in shrimp diets without compromising growth and FA profiles.     

Interactive effect of two methyl‐group sources on the growth and phospholipids’ content of kuruma shrimp Marsupenaeus japonicus Bate post‐larvae

Six micro‐bound diets were formulated to contain three levels of choline chloride (CC) (0.0, 0.6 and 1.2 g kg⁻¹) and 2 levels of methionine (Met) (0 and 15 g kg⁻¹). Soybean protein isolates (SPI) were used as the main protein source for its limited Met content. A significant (P < 0.05) interaction was determined between CC and Met on the survival (S %), weight gain (WG%), specific growth rate (SGR % day⁻¹), feed efficiency ratio (FER), protein efficiency ratio (PER) and phosphatidylcholine (PC), phoshphatidylethanolamine (PE) and Met contents of the whole body of shrimp. The shrimp group did not receive either supplemental CC or Met showed lower (P < 0.05) values of the above‐mentioned parameters than other shrimp groups fed with 0.6 and 1.2 g kg⁻¹ supplemental CC with or without Met supplementation. The present study showed that supplementation of 1.2 g kg⁻¹ CC in the diets could compensate shrimp post‐larvae with the needed methyl group when received Met‐deficient diets. The study also assumed that the biosynthesis of PC in the shrimp’s body can be achieved by the methylation of PE through the S‐adenosylmethionine (SAM) pathway and/or through the cytosine di‐phosphoryl (CDP) choline pathway directly from dietary choline.     

Effects of supplementation of coated crystalline amino acids on growth performance and body composition of juvenile kuruma shrimp Marsupenaeus japonicus

Supplementation of crystalline amino acids (CAA) in shrimp diets is unsuccessful in most cases. In the present study, various pre‐coated CAA were developed to minimize the leaching losses from diets in water and assessed using kuruma shrimp Marsupenaeus japonicus juveniles. Experimental diets were supplemented with coated CAA or non‐coated CAA to simulate the dietary amino acid profile to that found in the whole body protein of shrimp. The four diets contained CAA coated with either carboxymethyl cellulose (CMC) (diet 1), zein (diet 2), k‐carrageenan (diet 3) or agar (diet 4), respectively. Another four diets contained CMC‐coated CAA with the addition of either zein (diet 5), casein‐gelatin (diet 6), zein and k‐carrageenan (diet 7), or casein‐gelatin and k‐carrageenan (diet 8), respectively. The 8 weeks feeding trial demonstrated that the diets containing coated CAA gave significantly higher weight gains than diets containing non‐coated CAA, with the highest value for the diet 6. The diets containing coated CAA also yielded higher feed conversion efficiency, protein efficiency ratio, and specific growth rate than diets containing non‐coated CAA. Leaching trials suggested that shrimp growth performance was improved partly due to the retardation or reduction of CAA leaching by coating them with specific binders.     

Effects of water-soluble and fat-soluble choline sources on the performances of juvenile kuruma shrimp, Marsupenaeus japonicus Bate

The present study was undertaken to clarify the effect and interaction of choline chloride (CC) and phosphatidylcholine (PC), the water‐ and fat‐soluble sources of choline, on the performances of kuruma shrimp, Marsupenaeus japonicus juveniles using as response variables such as percent weight gain (%WG), specific growth rate (SGR), survival (SR%), feed efficiency ratio (FER), protein efficiency ratio (PER), proximate composition, fatty acids profile and free choline and PC contents in the whole body of kuruma shrimp. A 40‐day feeding trial was conducted as a 2 × 3 factorial experiment with six test diets containing three levels of CC (0.3, 0.6 and 1.2 g kg^?1) and two levels of soybean PC (SPC; 2 and 4 g kg^?1). Both of the choline sources (CC and SPC) showed a significant interaction (P<0.05) regarding all the evaluated response variables except for the proximate and fatty acid contents. Interestingly, there was a positive effect (P<0.05) of CC observed with increasing its level at the low level of SPC and vice versa. However, the magnitudes of the beneficial effect of CC in terms of the evaluated parameters were changed with the different dietary levels of SPC. Also, the inclusion of higher levels of SPC (4 g kg^?1) or CC (0.6 and 1.2 g kg^?1) diminished the effect of the other source.     

Effect of replacing fishmeal with canola meal on growth and nutrient utilization in kuruma shrimp Marsupenaeus japonicus (Bate)

A feeding trial was conducted to assess the nutritional values of canola meal as a substitute for fishmeal in diets for kuruma shrimp using five isocaloric diets (190 kJ kg^?1) prepared by replacing 0%, 10%, 20%, 30% and 40% of fishmeal protein with canola meal protein in CM0, CM10, CM20, CM30 and CM40 diets respectively. Triplicate groups of juveniles (0.19 g) were fed the respective diets for 60 days in tanks. At the end of trial, weight gain (%) and specific growth rate (% day^?1) were not significantly (P > 0.05) different among shrimp fed CM0, CM10 and CM20 diets. However, growth was significantly (P < 0.05) decreased in shrimp fed CM30 and CM40 diets. As with growth performance, feed intake, feed conversion ratio and protein efficiency ratio were also decreased (P < 0.05) in CM30 and CM40 groups. Whole body dry matter, methionine and proline were significantly decreased with canola meal substitution levels. In general, retention efficiency of protein and indispensible amino acids were decreased as canola meal increased in diets. It may be concluded that 20% fishmeal protein can be successfully replaced with canola meal, while supplementation of amino acids or blending with complementary proteins could facilitate higher fishmeal replacements in kuruma shrimp diets.     

Short-term responses of adult kuruma shrimp Marsupenaeus japonicus (Bate) to environmental salinity: osmotic regulation, oxygen consumption and ammonia excretion

The effects of salinity on haemolymph osmolality, oxygen consumption and ammonia excretion were investigated in adult kuruma shrimp, Marsupenaeus japonicus (Bate), at salinities of 20, 25, 30 and 35 g L^?1. Haemolymph osmolality showed a positive linear relationship (r²=0.9854) with medium osmolality. The isosmotic point calculated from this relationship was 1039 mOsm kg^?1, which corresponds to a salinity of approximately 35 g L^?1. The slope of the regression equation was very high (0.81), suggesting that M. japonicus adults are poor osmoregulators compared with the adults of other penaeids and to conspecific young. The difference between haemolymph and medium osmolality (D_OP) was lowest at 35 g L^?1 and highest at 20 g L^?1. Thus, the minimum D_OP coincided with the isosmotic point of the shrimp. The respiration rate was significantly lower at 30 g L^?1 than at the other salinities. Ammonia excretion rates were inversely related with salinity and therefore were minimal at 35 g L^?1. The results of this study suggest that the optimum salinity for adult kuruma shrimp is around 30–35 g L^?1 and that even minor (e.g. 5 g L^?1) deviations from this optimum cause significant physiological changes. Further, the observed increases in oxygen consumption and ammonia excretion during exposure to low salinities, which indicate higher energy expenditure and amino acid catabolism for osmoregulation, respectively, suggest that the growth efficiency of M. japonicus adults may be severely compromised by hypohaline water inflow into the rearing ponds.     

Vitamin C requirement of kuruma shrimp postlarvae, Marsupenaeus japonicus (Bate), using l-ascorbyl-2-monophosphate-Na/Ca

l ‐ascorbyl‐2‐monophosphate‐Na/Ca (AMP‐Na/Ca) was used as a vitamin C source to investigate the ascorbic acid (AsA) requirements on growth performance and stress resistance of the post‐larval kuruma shrimp, Marsupenaeus japonicus. Purified carrageenan‐microbound diets with six levels of AMP‐Na/Ca, AsA equivalent to 0, 20, 56, 87, 759 and 1697 mg kg^?1 diet were fed to triplicate groups of M. japonicus (mean initial weight 16±0.3 mg) for 30 days. The diets with AsA 0, 20 and 56 mg kg^?1 showed high cumulative mortality after 10 days of feeding. After the 30‐day trial, the shrimp fed these diets had significantly lower survival and weight gain (WG, %) than those fed the 87, 759 and 1697 mg AsA kg^?1 diets. Specific growth rate and individual dry weight showed the same pattern as WG (%). There were no significant differences in growth performance among the groups fed the AsA levels at 87, 759 and 1697 mg kg^?1 at the termination of feeding trial. Broken‐line regression analysis indicated that 91.8 mg AsA kg^?1 in the diet was the optimum for post‐larval shrimp. On the other hand, dietary level of more than 800 mg AsA kg^?1 was needed to ensure high resistance to stressful conditions such as osmotic and formalin stressors.     

Replacement of squid and krill meal by snail meal (Buccinum striatissimum) in practical diets for juvenile of kuruma shrimp (Marsupenaeus japonicus)

A 60‐day feeding trial was conducted to investigate the effects of substituting squid and krill meal with marine snails (Buccinum striatissimum) into the diets of juvenile kuruma shrimps (Marsupenaeus japonicus). Five experimental diets were formulated to contain varying levels of snail meal at 0%, 25%, 50%, 75% and 100% (D1, D2, D3, D4 and D5, respectively) and fed to juvenile kuruma shrimps (initial mean weight 0.27 ± 0.02 g). The results showed that weight gain, feed intake and specific growth rate were improved significantly in D4 and D5 groups when compared with D1 group (p < 0.05). Significant differences were not detected in survival rate among all shrimps fed diets containing several levels of snail meal (p > 0.05). Crude protein content of shrimps fed the control diet was significantly lower than other treatments (p < 0.05). Lipid content in shrimps fed 50% snail meal were significantly higher than the control while cholesterol content in shrimps fed 100% snail meal were significantly decreased and polyunsaturated fatty acid contents were significantly increased in shrimps fed 75%–100% snail meal (p < 0.05). These results suggest that supplementing snail meal for complete replacement of squid and krill meal can be done to improve juvenile kuruma shrimps’ growth and reducing their cholesterol levels.     

Dietary supplementation with Bacillus can improve the growth and survival of the kuruma shrimp Marsupenaeus japonicus in high-temperature environments

This study examined the effects of a commercial Bacillus complex used as probiotics on the performance of Marsupenaeus japonicus under high water temperature after 60 days of rearing. Growth and survival rate of M. japonicus were significantly increased in the probiotics-treated group (P < 0.05). Vibrio count and percentage of Vibrio in the intestinal tracts of M. japonicus reared with Bacillus were significantly lower than in other treatments (P < 0.05). The expressions of immune-related gene including prophenoloxidase, lysozyme, cytosolic manganese superoxide dismutase and hemocyanin subunit L in M. japonicus were significantly increased in the Bacillus-treated group (P < 0.05). The results indicated that Bacillus can be used as a diet supplement to minimize the damages caused by free radicals generated from insufficient oxygen metabolism due to the high-temperature stress, and to enhance immunity and activate the immune response levels. Our findings can improve the growth and survival rates of M. japonicus during the high-temperature farming period.     

Nutrient values of dietary ascorbic acid (l-ascorbyl-2-polyphosphate) on growth, survival and stress tolerance of larval shrimp, Litopenaeus vannamei

l -ascorbyl-2-polyphosphate (ApP) was used as a vitamin C source to investigate the ascorbic acid (AsA) requirements on growth performance and stress resistance of the larval white shrimp, Litopenaeus vannamei . Five isoenergetic and isonitrogenous fish meal-fish protein hydrolysate-based diets with five levels of ApP, AsA equivalent to 91.8, 188, 271, 360 and 436 mg kg⁻¹ diet were fed to triplicate groups of L. vannamei (mean initial wet weight 1 mg) for 32 days. The diet with AsA 91.8 mg kg⁻¹ showed high cumulative mortality after 10 days of feeding. After the 32-day trial, the shrimp that fed the diet had significantly lower survival and weight gain (WG, %) than those that fed 188, 271, 360 and 436 mg AsA kg⁻¹ diets. Specific growth rate (SGR, % day⁻¹) and final body wet weight (FBW, mg) showed the same pattern as WG (%). There were no significant differences in growth performance (FBW, WG and SGR) among the groups that fed 188, 271, 360 and 436 mg kg⁻¹ of AsA at the termination of feeding trial. Broken-line regression analysis on WG indicated that 191 mg AsA kg⁻¹ in the diet was the optimum for larval L. vannamei . On the contrary, dietary level of more than 360 mg AsA kg⁻¹ was needed to ensure high resistance to stressful conditions such as low dissolved oxygen stressors.     

Effects of dietary chitosan on growth,survival and stress tolerance of postlarval shrimp,Litopenaeus vannamei

The effect of chitosan, a polymer of glucosamine obtained by the deacetylation of chitin, on growth, survival and stress tolerance was studied in postlarval Litopenaeus vannamei. An experiment was performed with postlarval shrimp (mean initial wet weight 1.2 mg) fed five isoenergic and isonitrogenous diets containing five supplemented levels of chitosan (0, 0.5, 1, 2 and 4 g kg^?1 diet, respectively). The five compound diets (C0, C0.5, C1, C2 and C4) sustained shrimp growth throughout the experiment. Growth performance (final body weights; weight gain; SGR: specific growth rate) in shrimp fed diet C2 was significantly higher than that in shrimp fed diets C0, C0.5 and C1 (P < 0.05), diet C4 treatment provided intermediate growth result. The survival in shrimp fed diet C1 was significantly higher than that in shrimp fed C0 diet (P < 0.05), other diets treatments gave the intermediate survival results. No significant differences were found in growth and survival between diet C2 and C4 treatments. After 9 days of a stress tolerance test, survival in shrimp fed diets C1, C2 and C4 was significantly higher than that in shrimp fed diets C0 and C0.5. We concluded from this experiment that the incorporation of a moderate dietary chitosan was beneficial to the development of postlarval L. vannamei. Considering the effect of chitosan on both growth and survival of postlarval L. vannamei, second‐degree polynomial regression of SGR and survival indicated optimum supplement of dietary chitosan at 2.67 and 2.13 g kg^?1, respectively, so the level of chitosan supplemented in the diet should be between 2.13 and 2.67 g kg^?1.     

Interactive effects of dietary vitamin C and phospholipid in micro‐bound diet for growth,survival, and stress resistance of larval red sea bream,Pagrus major

This study was conducted to examine the effects of dietary ascorbic acid (AsA) and phospholipid (PL) and their interaction on growth, survival, and stress resistance in red sea bream larvae. Twenty‐six days old red sea bream were fed nine micro‐bound diets supplemented three levels of AsA (0, 800 and 1600 mg kg^?1 diet) and PL (0, 20 and 40 g kg^?1 diet) for 15 days. Dietary AsA and PL were both significant factors on survival rates. There was also an interaction between dietary AsA and PL on survival rate (P < 0.05). The larvae fed 800 or 1600 mg kg^?1 AsA with 40 g kg^?1 PL diets showed the highest survival rate, with values similar to those of the live‐food supplemented group. Stress resistance against low salinity exposure significantly increased with increased dietary level of AsA and PL. However, significant interaction of AsA and PL was not detected. The larvae fed 1600 mg kg^?1 AsA with 40 g kg^?1 PL diet showed the highest stress resistance among all diets, but it was not significantly different than that of larvae fed 800 mg kg^?1 AsA with 40 g kg^?1 PL diet. This study clearly demonstrated that combined use of AsA and PL can improve survival of 26–40 days posthatching red sea bream larvae. Moreover, the present study suggested that 800 mg kg^?1 AsA with 40 g kg^?1 PL in diet was needed for producing high quality seedling under the stressful conditions.     

Effect of choline chloride as an osmoregulator as well as its role in growth and the biochemical content of postlarval kuruma shrimp; Marsupenaeus japonicus (Bate)

A 45‐day feeding experiment was conducted to examine two levels of dietary choline chloride (CC) [0.6 and 1.2 g kg^?1 CC] against control (0.0 g kg^?1 CC) on the biological, biochemical composition and stress tolerance of postlarval (PL‐20) Marsupenaeus japonicus. The results showed improvements in some biochemical contents such as phosphatidylcholine (PC) and methionine (Met) as 0.6 and 1.2 g kg^?1 dietary CC significantly (P < 0.05) increased both PC and Met contents of shrimp carcass more than those of the control group. The results also showed that supplemental (0.6 g kg^?1) choline could improve (P < 0.05) PC/PE (phosphatidylethanolamine) ratio. Growth parameters of the shrimp group that received 0.6/1.2 g kg^?1 dietary CC showed a significant (P < 0.05) improvement. In case of osmotic stress test, 0.6 g kg^?1 supplemented CC level showed significantly (P < 0.05) longer LT‐50 (the lethal time needed to kill half of the population) than the other shrimp groups that received 0.0 and 1.2 g kg^?1 dietary CC levels. This study showed the efficacy of 0.6 g kg^?1 dietary choline to improve the osmotic tolerance of kuruma shrimp as well as growth, survival and biochemical contents of postlarval kuruma shrimp.     

Effects of dietary supplementation of a commercial prebiotic Previda® on survival,growth, immune responses and gut microbiota of Pacific white shrimp,Litopenaeus vannamei

A 35‐day feeding trial was conducted to evaluate growth, bacterial populations of the gastrointestinal tract (GIT) and immune responses of Litopenaeus vannamei fed diets containing the commercial prebiotic Previda^®. Diets were formulated to contain Previda^® at 0, 0.2, 0.5, 1.0 or 1.6 g kg^?1 by weight. At the end of the study, differences in weight gain and survival among treatments were not significant (P > 0.05), but denaturing gradient gel electrophoresis analysis revealed that the microbial communities in the GIT changed significantly with the inclusion of dietary Previda^® at different levels. Previda^® was therefore able to selectively modify the microbial communities in the shrimp's GIT. Although individual bacterial species were not identified, enteric populations in shrimp fed the prebiotic at similar levels of inclusion were genetically similar. In addition, shrimp fed Previda^® at 1.6 g kg^?1 responded significantly (P < 0.05) better immunologically with respect to hemocyte phagocytic capacity, haemolymph protein, hyaline cell counts and haemolymph glucose compared with shrimp fed the basal diet. Although shrimp were not exposed to virulent pathogens in this study, the observed upregulation of some of imm‐une responses upon prebiotic supplementation indicates that an improved outcome of such challenges may be anticipated in Previda^®‐fed shrimp under commercial conditions.     

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.     

Experimental horizontal transmission of Enterocytozoon hepatopenaei in post‐larvae of whiteleg shrimp,Litopenaeus vannamei

Enterocytozoon hepatopenaei (EHP) is a microsporidian parasite that causes hepatopancreatic microsporidiosis (HPM) in penaeid shrimp. HPM was observed in several countries, including Thailand and India; it has become a prominent pathogen in shrimp culture. Based on observations on EHP infection in the wild, the route of transmission has been hypothesized. Identification of artificial EHP infection procedures can facilitate our understanding of EHP transmission. Experimental transmission of EHP was attempted using the immersion and oral infections of infection. In the immersion mode, post‐larvae (PL) were exposed to an EHP tissue homogenate (0.2%) by immersion for 48 hr. Experimental samples were collected at various time points, and infection was confirmed using polymerase chain reaction, haematoxylin and eosin staining, transmission electron microscopy and modified trichrome staining. All test results revealed successful EHP transmission. Similar results were obtained through oral infection (oral infection). Innate immune gene expression patterns during infection were analysed; prophenoloxidase, crustin and superoxide dismutase were upregulated at 6, 6 and 48 hr post‐challenge, respectively. Experimental infection procedures facilitate the development of diagnostic and prevention strategies. This is the first study demonstrating the experimental transmission of EHP in shrimp PL.     

Effects of dietary nucleotides on growth,survival and metabolic response in whiteleg shrimp,Litopenaeus vannamei against ammonia stress condition

This trial was conducted to evaluate the effects of nucleotides on growth of whiteleg shrimp, Litopenaeus vannamei, and the survival and metabolic responses to ammonia stress test. Experimental diets were as follows: low fish meal diet (LFMD), and four LFMD test diets, each supplemented with 0.1% guanosine monophosphate (GMP), 0.1% inosine monophosphate (IMP), 0.1% mixture of GMP and IMP and 0.1% mixture of GMP, IMP, uridine monophosphate (UMP) and cytidine monophosphate (CMP). The shrimp specimens (initial body weight: 0.99 ± 0.01 g) were randomly allocated into five groups and fed four times daily for 8‐weeks. After the trial, final body weight was recorded and haemolymph was withdrawn for haematological analysis. The shrimp was then challenged with 70 mg/L ammonia (LC50) for 10 days. Survival and haemolymph of the shrimp were taken after exposure to ammonia. The highest growth performance was observed in the shrimp fed diet supplemented with GMP (p < .05), while survival was not influenced by the test diets in the feeding trial. In the ammonia challenge test, the highest survival was observed in the shrimp fed GMP supplemented diet compared to others. The plasma protein, glucose and cholesterol levels increased in all the treatments while triglycerides level decreased post challenge. Cortisol level recovered at day 10th after the challenge. Shrimps fed with nucleotides diets showed higher protein and glucose level compared to control groups post challenge. In general, nucleotides supplemented in the diet enhanced growth, improved stress resistance while modulating the haemolymph metabolites in L. vannamei under ammonia stress.