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 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.     

Effects of low salinity exposure on immunological,physiological and growth performance in Litopenaeus vannamei

Growth, immunological and physiological parameters of white shrimp Litopenaeus vannamei reared at different salinity levels (1, 10, 15, 25 and 35 g/L) at stocking density of 214 shrimp/m³ were examined at 1, 30 and 63 days. Results showed that the total haemocyte count (THC) of shrimp decreased with time at all salinity levels, indicating a potential reduction in the resistance of shrimp against pathogens, since a low value of THC indicates a perturbation of the immune system. Glucose and protein values observed in the haemolymph throughout the study indicate that shrimp adapted well to low salinities (1, 10 and 15 g/L). Although of those shrimp reared at 10 g/L only 83.3% survived, at this salinity, shrimp depicted a higher glucose concentration in haemolymph at the beginning and end of the study.     

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.     

Evaluating the effects of dietary resistant starch on growth,digestion, metabolism,immunity and resistance to ammonia stress in Litopenaeus vannamei

Resistant starch (RS) is considered to be a good alternative to antibiotics. A 56‐day feeding trial was followed by induction of acute ammonia stress to evaluate the effects of dietary RS on growth, digestion, metabolism, immunity and resistance to ammonia stress in Litopenaeus vannamei. The four diets used differed only in RS content: 0 g/kg (Control), 10 g/kg (RS1), 30 g/kg (RS2) and 50 g/kg (RS3). The results showed that dietary RS improved the growth and the survival rates of shrimp exposed to ammonia stress. At 56 days, the activity of digestive and metabolic enzymes (amylase, lipase, pepsin, hexokinase and pyruvate kinase) was increased in the three RS groups, while trypsin activity only increased in the RS1 and RS2 groups; immune‐related parameters (the total antioxidant capacity; the activity of superoxide dismutase, total nitric oxide synthase and phenoloxidase; and the expression of heat‐shock protein 70, thioredoxin, prophenoloxidase, lysozyme and toll gene) were increased in the three RS groups. After exposure to ammonia stress, all the selected immune parameters of the three RS groups were higher than those of the control group at 24 hr. Therefore, dietary RS improved the growth, digestion, metabolism, immunity and resistance to ammonia stress in L. vannamei.     

Immune responses of Litopenaeus vannamei to non‐ionic ammonia stress: a comparative study on shrimps in freshwater and seawater conditions

To investigate the effect of non‐ionic ammonia (NH₃‐N) stress (0.1 and 0.5 mg L^?1) on the immunity of Litopenaeus vannamei cultured in long‐term freshwater, the total haemocyte count (THC), the activity of phenoloxidase (PO), nitric oxide synthase (NOS), superoxidase dismutase (SOD) and the content of malondialdehyde (MDA) were determined and further compared with those of seawater shrimps. The results showed that NH₃‐N stress significantly reduced THC and the activity of PO and SOD (P < 0.05). Under 0.1 mg L^?1 NH₃‐N stress, NOS activity increased first and then decreased significantly, while it dropped dramatically under 0.5 mg L^?1 NH₃‐N stress (P < 0.05). During NH₃‐N stress, MDA content increased continuously, and the MDA content in hepatopancreas of freshwater shrimps was higher than that of seawater shrimps. It was concluded that NH₃‐N stress significantly influenced the non‐specific immunity and could also upset the balance of antioxidant system of L. vannamei in both freshwater and seawater shrimps. Compared with in seawater, the shrimps in freshwater were more vulnerable to NH₃‐N stress because of higher lipid peroxidation and lower immunity.     

Effects of dietary supplementation of Astragalus membranaceus,Codonopsis pilosula,and Glycyrrhiza uralensis extract mixture on growth performance,haematological parameters and hepatopancreatic performance in juvenile Pacific white shrimp (Litopenaeus vannamei)

Pacific white shrimp is the major farmed shrimp species in the world. It is known to be very sensitive to the environmental and management changes such as intensification, which is one of the primary necessities to increase shrimp production, but represents a stressful condition, and needs to be managed properly to diminish its negative effects in aquaculture. In this study, juvenile Pacific white shrimp were fed diets supplementing with equal quantities of Astragalus membranaceus, Codonopsis pilosula and Glycyrrhiza uralensis extracts in different concentrations (0, 1.0, 2.0 and 3.0 g/kg) for 8 weeks. We subsequently estimated the effects of supplemented diets on growth performance, haematological parameters, and histological changes of hepatopancreas. Results revealed that the supplemented mixture had no benefits on growth and survival rate, but had favourable effects on haemocyte count, number of granulocytes, and hemocyanin concentration in the haemolymph (p > 0.05). We observed that R cells and B cells were increased in hepatopancreatic tubules of shrimps fed on supplemented diets. Besides, a diet containing 2.0 g/kg of extract mixture maintained the decreased levels of alanine aminotransferase (p < 0.05) and aspartate aminotransferase (p > 0.05). Among the used concentrations, 2.0 g/kg seemed to be the most suitable concentration.     

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.     

Effects of Lactobacillus pentosus on the growth performance,digestive enzyme and disease resistance of white shrimp,Litopenaeus vannamei (Boone, 1931)

The objective of this study was to evaluate the probiotic properties of lactic acid bacteria (LAB) strains isolated from digestive tract of white shrimp Litopenaeus vannamei. Eighteen LAB colonies were isolated and one bacterium was found capable of producing three extracellular enzymes (protease, cellulose and lipase) simultaneously and exhibited antagonistic activity against shrimp pathogens (Vibrio vulnificus, V. rotiferianus and V. campbellii). The putative probiotic strain AS13 was identified as Lactobacillus pentosus based on 16S rRNA sequencing. The L. vannamei were fed diet containing 0 (control), 10⁶, 10⁷ and 10⁸ CFU g^?1 bacterial cells of AS13 for 28 days. The results showed that supplementation of L. pentosus significantly improved the growth performance and feed utilization in the treated groups over the control. Similarly, digestive enzyme activities were elevated in the intestines of treated groups. Moreover, feeding of supplemented diets containing AS13 significantly reduced the mortality rate caused by pathogenic Vibrio species (V. vulnificus, V. rotiferianus and V. campbellii). Our results indicated L. pentosus AS13 addition at 10⁷ CFU g^?1 can effectively enhance the growth performance, feed utilization, digestive enzymes and disease resistance of L. vannamei in the laboratory condition.     

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.     

Effects of dietary taurine,carnitine and cholesterol supplementation on growth performance and immunological status of Litopenaeus vannamei under cold exposure

The aim of this study was to investigate effects of dietary feed additives on growth and immunological status of Litopenaeus vannamei under cold exposure. Four experimental diets with 0.2% taurine, 0.01% carnitine, 0.1% cholesterol supplementation and control were fed to L. vannamei (IBW 0.86 ± 0.03 g) for 30 days at 30 ± 1°C before water temperature was dropped to 16°C with a rate of ?4°C h^?1. Growth performance parameters including weight gain rate (WGR), feed conversion ratio (FCR) and survival rate before the cold exposure, as well as immunological parameters related to haemocytes including total haemocyte count (THC), percentages of hyaline cells (SC%), granular cells and semigranular cells (SC + GC%), non‐specific esterase (NSE) activity and apoptotic cell ratio before and after the cold exposure were determined. The results showed that: (1) WGR, FCE and survival rate were not significantly different among groups before the cold exposure. Most of immunological parameters did not differ either, except the apoptotic cell ratio of carnitine and cholesterol groups was significantly lower than that of control and taurine groups (P < 0.05). (2) After the cold exposure, THC was decreased by 48.1% (P < 0.05), 54.1% (P < 0.05), 30.1% and 2.2%; NSE activity was decreased by 31.7% (P < 0.05), 21.4%, 7.0% and 3.5% in control, taurine, carnitine and cholesterol respectively. (3) Conversely, ROS production was increased by 23.8% (P < 0.05), 15.1%, 7.2% and 13.0%; apoptotic cell ratio was increased by 10.4%, 12.8%, 6.4% and 7.2% in control, taurine, carnitine and cholesterol respectively. (4) An increase in HC%, decrease in SC + GC% in all groups was observed after cold exposure, and was significant in cholesterol group. This study showed that cholesterol, carnitine and taurine affected immunological status of L. vannamei under cold exposure to different degrees, possibly by modulating some immunological parameters, as discussed. Present study provides theoretical data for nutritional strategies to enhance cold resistance of shrimp.     

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 juvenile white shrimp,Litopenaeus vannamei: Growth,feed utilization,antioxidant enzymes activities and fatty acid compositions

Present study investigates the effects of n‐3 high‐unsaturated fatty acid (n‐3HUFA) levels on growth performance, antioxidant enzymes activities and fatty acid compositions of juvenile Litopenaeus vannamei. These represented seven iso‐nitrogenous and iso‐lipidic diets. Analysed n‐3HUFA concentrations were 0.16% (control), 0.48%, 0.74%, 1.39%, 2.39%, 2.92% and 3.44% respectively. A total of 840 juvenile L. vannamei were randomly stocked into 21 0.5 m³ tanks for 56 days. A significant increase (p < 0.05) was observed from 0.16% to 0.74% n‐3HUFA and a decrease when n‐3HUFA was above these levels in weight gain rate (WGR) and specific growth rate. Total cholesterol, triglyceride and low‐density lipoprotein in serum showed a significant decrease, high‐density lipoprotein showed a significant increase (p < 0.05). Phenoloxidase activity in serum and sodium‐potassium adenosine triphosphatase activity in gill were significantly affected by dietary n‐3HUFA (p < 0.05), both of them showing a downward trend after upward. Malic dehydrogenase and superoxide dismutase activities in serum were also significantly affected by dietary n‐3HUFA (p < 0.05), which rose first and then decreased in general, both of them have a maximum in 2.39% group. No significant differences of the activities of aspartate transaminase and alanine transaminase were observed among all groups (p > 0.05). With dietary n‐3HUFA increase, both ∑HUFA and n‐3HUFA contents gradually increased in hepatopancreas and tail muscles (p < 0.05). Based on broken‐line regression analysis of WGR, the optimal n‐3HUFA requirement is 0.89% for juvenile L. vannamei with initial weight of 0.50 ± 0.01 g.     

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

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

Effects of dietary manganese source and supplemental levels on growth performance,antioxidant enzymes activities,tissue Mn concentrations and cytosolic manganese superoxide dismutase (cMnSOD) mRNA expression level of juvenile Litopenaeus vannamei

This experiment was conducted to study the effects of different forms and levels of manganese (Mn) on the growth performance, antioxidant activities, tissue Mn content and cytosolic manganese superoxide dismutase (cMnSOD) gene expression of Litopenaeus vannamei. Treatments consisted of 0, 10, 20, 30, 40 and 60 mg Mn kg^?1 from manganese sulphate (Mn‐S) and manganese methionine (Mn‐Met), providing the actual dietary value of 5.17, 15.62, 25.55, 34.22, 44.48 and 67.90 mg Mn kg^?1 Mn‐S, and 5.17, 15.71, 25.36, 35.86, 45.16 and 65.06 mg Mn kg^?1 Mn‐Met, respectively. Each diet was fed to triplicate groups of L. vannamei (initial body weight: 1.925 ± 0.002 g) in a recirculated fresh water rearing system for 8 weeks. Weight gain rate (WGR) increased in prawns provided with from 25.55 to 44.48 mg Mn kg^?1 Mn‐S and 15.71 to 45.16 mg Mn kg^?1 Mn‐Met and then declined above these levels. The lowest protein efficiency ratio (PER) and the highest feed conversion rate (FCR) were observed in prawns fed the control diet (P < 0.05) and showed no significant differences among other treatments (P > 0.05). Survival rate (SR) was not affected by the dietary treatments (P > 0.05). Total SOD and Mn‐SOD activities were higher in the hepatopancreas of prawns fed with Mn‐supplemented diets from 15.71 to 44.48 mg Mn kg^?1 (P < 0.05). On the contrary, malondialdehyde (MDA) content was lower in the hepatopancreas of prawns fed the basal diet (P < 0.05). Mn concentrations in the hepatopancreas and muscles increased with increasing levels of dietary Mn supplementation. Moreover, Mn accumulation was lower in the muscle than in the hepatopancreas of the prawns. The mRNA expression of cMnSOD gene in the hepatopancreas of prawns was upregulated with increasing dietary Mn levels of Mn‐S from 25.55 to 44.48 mg Mn kg^?1, Mn‐Met from 15.71 to 45.16 mg Mn kg^?1 and then plateaued above these levels. Broken‐line regression analysis of WGR indicated that the optimal dietary Mn requirements for juvenile L. vannamei were 32.26 mg Mn kg^?1 Mn‐S and 23.90 mg Mn kg^?1 Mn‐Met, respectively.     

Effects of crude extracellular protein and crude intracellular polysaccharides of Vibrio alginolyticus on the growth,energy metabolism regulation and WSSV resistance of Litopenaeus vannamei

This study aimed to investigate the effects of adding crude extracts of the extracellular protein (Ex‐Pro) and intracellular polysaccharides (In‐Poly) of Vibrio alginolyticus to diets, at a dosage of 10 g/kg, on the growth, energy metabolism and WSSV (White Spot Syndrome Virus) resistance of Litopenaeus vannamei (initial weight, 0.88 ± 0.04 g/shrimp). Growth and survival rate were not significantly affected by the crude extracts (p > 0.05). Shrimp fed Ex‐Pro crude extracts showed higher succinate dehydrogenase (SDH) activity in the hepatopancreas and muscle but lower lactate dehydrogenase (LDH) activity in the muscle (p < 0.05). The activities of hexokinase (HK), pyruvate kinase (PK), LDH and SDH in the hepatopancreas and the activity of SDH in the muscle were significantly increased by feeding In‐Poly crude extracts (p < 0.05). In contrast, the content of fatty acid synthase (FAS) in the muscle was significantly reduced by the crude extracts (p < 0.05). The contents of glucose and triglyceride and the activity of the electron transport system in the hepatopancreas were significantly increased by the crude extracts (p < 0.05), and the WSSV resistance of the shrimp was increased. These results indicated that the Ex‐Pro and In‐Poly crude extracts of V. alginolyticus could affect energy metabolism, and there was a correlation between WSSV resistance and energy metabolism in L. vannamei.     

Administration of probiotic Bacillus licheniformis induces growth,immune and antioxidant enzyme activities,gut microbiota assembly and resistance to Vibrio parahaemolyticus in Litopenaeus vannamei

This study aimed to elucidate the effects of dietary Bacillus licheniformis ATCC 11946 (BL) on the growth, immune and antioxidant activities, intestinal morphology and microbiota, and susceptibility to Vibrio parahaemolyticus (VP_AHPND) in juvenile pacific white shrimp, Litopenaeus vannamei (LV). Juvenile LV (initial weight = 0.63 ± 0.001) were fed diets containing varying BL concentrations (0 (BL0), 10⁶ (BL1), 10⁷ (BL2) and 10⁸ (BL3) CFU/g feed) for 8 weeks. Growth performance, immune and antioxidant enzyme activities, and intestinal morphology significantly improved in the probiotic‐treated groups than the untreated. Regardless of the treatment group, the two dominant phyla were Proteobacteria and Bacteroidetes, whereas the two dominant genera were Ruegeria and Vibrio. Increasing inclusion of probiotics in diets led to significant increase in beneficial bacterial genera (Ruegeria, and Pseudoalteromonas) and a significant decrease in some known opportunistic pathogens (Vibrio, Tenacibaculum, Photobacterium, Kangiella and Spongiimonas) with the BL3 group witnessing the best. A 7‐day challenge study with VP_AHPND showed significantly high protection in the probiotic‐treated groups, with the BL3, BL2 and BL1 obtaining 51%, 34% and 22% relative percentage survival, respectively. In conclusion, BL at 10⁸ CFU/g feed should be used to help in shrimp production since it attained the most significant improvement.     

Effect of dietary vitamin B6 on growth,feed utilization,health and non‐specific immune of juvenile Pacific white shrimp,Litopenaeus vannamei

Six diets were formulated with vitamin B₆ levels (2.6, 32.7, 54.8, 90.7, 119.6 and 247.4 mg kg⁻¹, dry diet) to determine the requirement for juvenile Pacific white shrimp, Litopenaeus vannamei. Triplicate groups of 40 juvenile shrimp (approximately 1.0 g) were provided four times each day to apparent satiation (8 weeks). Weight gain (WG), specific growth rate, feeding efficiency, protein efficiency ratio (PER) and protein productive value of the shrimp were significantly influenced by the vitamin B₆ levels. No significant differences in whole‐body and muscle composition, except for dry matter and protein contents in whole body. Vitamin B₆ concentration in the hepatopancreas significantly increased with the dietary vitamin B₆ level increasing from 2.6 to 32.7 mg kg⁻¹. High‐density lipoprotein cholesterol in the haemolymph improved with the dietary vitamin B₆ levels increasing from 2.6 to 90.7 mg kg⁻¹ diet and no significant differences in low‐density lipoprotein cholesterol, cholesterol, glucose and total protein concentrations. Aspartate aminotransferase, alanine aminotransferase, superoxide dismutase, catalase and lysozyme in the haemolymph were significantly influenced by dietary vitamin B₆ levels. The optimal dietary vitamin B₆ requirements estimated using a two‐slope broken‐line model based on WG and SGR and an exponential model based on the vitamin B₆ concentration in the hepatopancreas were 110.39, 110.08 and 167.5 mg kg⁻¹, respectively.     

Potential role of LvDscam in specific immune response of Litopenaeus vannamei against white spot syndrome virus by oral delivery of VP28 using Bacillus subtilis

Envelope protein VP28 has been suggested as a candidate vaccine component to evoke a better protection against white spot syndrome virus (WSSV). We have reported that Bacillus subtilis spores harbouring VP28 (rVP28‐bs) can specifically protect shrimp against WSSV. However, the mechanism that supports the production of unique molecules induced by rVP28‐bs to trigger specific immunity is originally unknown. It has recently been suggested that Dscam (Down syndrome cell adhesion molecule) plays an essential role in the alternative adaptive immunity of invertebrates. In this study, we compared the diversity of Litopenaeus vannamei Dscam (LvDscam) variable regions by different antigens immunization. A total of 13, 15 and 11 expressed alternative sequences were identified for N‐terminal Ig2, N‐terminal Ig3 and the entire Ig7 domain, respectively. More than half of the unique variants (16 out of 22) were found in the Ig2/Ig3 domains. Further analysis of the interaction between VP28 and unique Ig2/Ig3 variants was confirmed by both yeast two‐hybrid and GST pull‐down approach. We also found that the percentage of haemocytes phagocytosing WSSV was significantly higher (P < 0.001) in the shrimp injected with control‐siRNA (43.8 ± 2.2) than those with Dscam‐siRNA (11.3 ± 5.4) in the rVP28‐bs groups. With Dscam‐siRNA injection, survivorship significantly decreased (P < 0.001) in the rVP28‐bs group after WSSV challenge. Our data suggested that LvDscam‐mediated pathway may be involved in the specific immune response of shrimp against WSSV induced by rVP28‐bs.