Dietary administration of soybean isoflavones enhances the immunity of white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus

The growth response, total hemocyte count (THC), respiratory burst (release of superoxide anion), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, nitric oxide synthase (NOS) activity and resistance to the pathogen Vibrio alginolyticus were measured in white shrimp (Litopenaeus vannamei) which had been fed diets supplemented with soybean isoflavones (SI) at 0, 5, 10, 20, 40 mg kg^?1 feed for 8 weeks. Dietary SI had no significant influence on survival rate of the shrimp. Significantly greater SGR was observed in treatments with 10, 20 and 40 mg kg^?1 SI supplement than that in control (P < 0.05). Shrimp fed a diet with 20 mg kg^?1 SI showed significant increase in THC, PO activity, respiratory burst activity, SOD activity as well as NOS activity (P < 0.05). Increased THC, respiratory burst activity, SOD activity and NOS activity were also noticed in shrimp fed SI at 10 or 40 mg kg^?1 (P < 0.05). Significant lower cumulative mortality after challenge was observed in shrimp fed diets with SI supplement (10.0–13.3%) than that in control group (36.7%) (P < 0.05). It is therefore concluded that oral administration of SI at 20 mg kg^?1 for 8 weeks could enhance the immune ability of shrimp and improve production in shrimp farming.     

Effects of continuous and alternate administration of β‐glucan and mannan‐oligosaccharide on the growth,immunity and resistance against Vibrio splendidus of sea cucumber Apostichopus japonicus (Selenka)

A 4‐week growth trial was conducted to compare the effects of different feeding strategies of dietary immunostimulants on the growth, immunity and resistance against Vibrio splendidus of sea cucumbers Apostichopus japonicus (Selenka). Six feeding strategies were set, including feeding immunostimulants‐free diet continuously (control), feeding dietary β‐glucan (1.25 g kg^?1 diet) continuously, feeding dietary mannan‐oligosaccharides (MOS; 2.00 g kg^?1 diet) continuously, feeding β‐glucan 2 days followed by MOS 5 days alternately, feeding β‐glucan 5 days followed by MOS 2 days alternately and feeding β‐glucan 7 days followed by MOS 7 days alternately. The sea cucumbers fed immunostimulants showed higher specific growth rate (SGR) and lower cumulative mortality than control (P < 0.05). When sea cucumbers were fed with β‐glucan continuously, total coelomocytes counts and superoxide anion were significantly higher than control on the 4th day (P < 0.05). However, these two immune parameters were not significantly higher than those in control after the 18th day (P > 0.05). While sea cucumbers continuously fed MOS, these two immune parameters were not significantly higher than control until the 15th day. All immune parameters of the sea cucumbers fed with β‐glucan and MOS alternately were significantly higher than those in control during the experiment (P < 0.05). The sea cucumbers fed with β‐glucan 7 days followed MOS 7 days alternately showed the highest SGR and second lowest cumulative mortality. It was suggested that this feeding strategy is most suitable for sea cucumbers.     

Effects of dietary supplementation with β‐glucan and synbiotics on growth,haemolymph chemistry,and intestinal microbiota and morphology in the Pacific white shrimp

The goal of this study was to investigate the effects of dietary supplementation with β‐glucan and microencapsulated probiotics (Bacillus subtilis or Pediococcus acidilactici) on growth performance, body composition, haemolymph constituents, and intestinal morphology and microbiota of the Pacific white shrimp Litopenaeus vannamei. Four treatment diets [basal diet (C), β‐glucan‐containing diet (β‐glu), β‐glucan plus B. subtilis‐containing diet (β‐glu+Bs), and β‐glucan plus P. acidilactici‐containing diet (β‐glu+Pa)] were fed to L. vannamei for 90 days. Shrimp fed the β‐glu and β‐glu+Pa diets exhibited similar growth performance and body protein content, which were significantly higher than those of shrimp fed the control diet (P < 0.05). No significant differences in haemolymph triglyceride, cholesterol, protein, haemolymph urea nitrogen or chloride were detected among the experimental diets. However, dietary β‐glucan alone increased the haemolymph glucose level and osmolarity (P < 0.05). Synbiotic supplementation had greater effects on intestinal microbiota and morphology than dietary β‐glucan alone. For example, β‐glu+Bs increased the number of intestinal lactic acid bacteria and decreased the number of Vibrio spp. (P < 0.05), and β‐glu+Pa increased the height of intestinal villi.     

Immunomodulatory Effects of Dietary β‐glucan and Vitamin C in Nile Tilapia,Oreochromis niloticus L., Subjected to Cold‐induced Stress or Bacterial Challenge

To determine the dietary β‐glucan and vitamin C levels that would enable the fish to cope with different stress, 252 tilapia (16.86 ± 0.24 g) were distributed in thirty six 250‐L aquaria and fed experimental diets for 60 d. Diets were supplemented with β‐glucan (0.1, 0.2, 0.4, and 0.8%) and vitamin C (400 and 600 mg/kg diet) and distributed in a factorial design that included a control diet (0 β‐glucan/125 mg vitamin C/kg). After growth performance analyses, a group of 108 fish were subjected to cold‐induced stress and another to Aeromonas hydrophila infection. Hematological and immunological parameters were determined before and after the stress. Growth performance was not affected. Red blood cells, hematocrit, mean corpuscular volume, total plasma protein, albumin : globulin ratio and leukocytes, lymphocytes, neutrophils, and monocytes were affected (P < 0.05) by stress and/or diets. The control diet resulted in higher post‐infection cortisol (P < 0.05). Liver vitamin C was directly proportional to dietary vitamin C (P < 0.05). The production of hydrogen peroxide and nitric oxide was also affected by diets and/or bacterial challenge (P < 0.05). In total, 0.1–0.2% β‐glucan and 600 mg/kg vitamin C increased fish resistance to stress and 0.8% β‐glucan resulted in reduced immune responses regardless of the vitamin C supplementation level.     

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.     

Use of Diet Crossover to Determine the Effects of β‐glucan Supplementation on Immunity and Growth of Nile Tilapia,Oreochromis niloticus

Juvenile Nile tilapia were fed either a basal (control) diet (n = 6 aquaria) or a diet supplemented with 1 g/kg β‐glucan (n = 24 aquaria) for 4 wk. At the end of this period, fish receiving β‐glucan were continued on the same diet (n = 12 aquaria) or switched to the control diet (n = 12 aquaria) for 2 wk. After 6 wk, tilapia continuously fed the β‐glucan supplemented diets had improved weight gain and feed efficiency than those fed the control diet uninterrupted or switched from the β‐glucan diet to the control after 4 wk. Feeding tilapia β‐glucan for 4 wk and then switching to the basal diet for 2 wk caused a significant increase in the respiratory burst of polymorphonuclear lymphocytes (17.77 × 10³ units/1000 white blood cells [WBC]) compared to catfish fed the control diet (13.50 × 10³ units/1000 WBC) or the β‐glucan diet continuously (13.57 × 10³ units/1000 WBC), but other immune parameters were unaffected. Tilapia were then challenged with Streptococcus iniae. The two groups were divided again (n = 6 aquaria) postchallenge and continued on the same diet or switched to the other diet (β‐glucan or control) for another 3 wk. No differences in survival to S. iniae infection occurred between dietary groups.     

Effect of dietary β‐1,3‐glucan on the immune response of Litopenaeus vannamei exposed to nitrite‐N

β‐1,3‐Glucan with 1.0 g kg^?1 was supplemented to a basal diet [control (C)] with different feeding schedules: permanently β‐1,3‐glucan diet (BG) and 14 days BG–14 days control diet (C + BG). Growth and immunological responses [respiratory burst (RB), superoxide dismutase (SOD), catalase (CAT), lysozyme and total protein] in haemolymph, hepatopancreas and muscle of Litopenaeus vannamei were recorded after 84‐day feeding and exposure to nitrite‐N (20 mg L^?1) for 120 h. β‐1,3‐Glucan administration did not affect growth performance. However, as compared with control, elevated CAT and lysozyme activities were seen in haemolymph of both BG groups, while significantly higher activities of SOD, lysozyme and RB in hepatopancreas, and higher activities of CAT and lysozyme in muscle were only seen in C + BG group. After nitrite‐N stress, significantly higher haemolymph protein, and hepatopancreas activities of lysozyme and RB were observed in both BG groups, but significantly higher activities of haemolymph SOD was only seen in C + BG group. The mortality in groups BG and C + BG was significantly lower than that in group C, but C + BG group showed a trend of higher nitrite‐N resistance compared with BG group. Considering dietary cost and immunostimulatory effects, the feeding schedule with 14 days BG–14 days control diet is more recommended.     

Enhancement of the innate immune system and disease‐resistant activity in Cyprinus carpio by oral administration of β‐glucan and whole cell yeast

The effects of dietary β‐ (1,3) glucan and whole cell yeast (Sacharomyces uvarum) on the immune response and disease resistance to Aeromonas hydrophila were investigated in Cyprinus carpio. β‐(1,3) glucan was extracted from the yeast. Both β‐(1,3) glucan and whole yeast were incorporated into the diet at 1% level and fed to common carp C. carpio for a period of 60 days. Control and treated fish were exposed to A. hydrophila on the 30th and the 60th day of the experimental period. Dietary supplementation of glucan significantly increased the white blood cell count in fish on the 60th day (2.91±0.04 × 10⁴), and the highest nuetrophil nitro blue tetrazolium (NBT) activity was also observed in glucan‐fed fish (30th day). A consistent increase in neutrophil (NBT) activity was also observed in whole cell fed fish until the end of the experiment. Similarly, β‐(1,3) glucan and whole cell yeast enhanced the serum lysozyme activity from the 15th day onwards but higher activity was reported on the 30th day in glucan and the 60th day in whole cell yeast‐fed fish. Suplementation of β‐(1,3) glucan protected the fish from A. hydrophila infection. Nearly 75–80% of the fish survived pathogen exposure (relative percentage survival). However, only 54–60% survival was observed in the whole cell‐fed fish. β‐(1,3) glucan and whole cell yeast protect the fish from pathogens by enhancing the cellular and humoral immune response in C. carpio.     

The effect of β‐1,3‐glucan derived from Euglena gracilis (Algamune™) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.)

Algamune^? is a commercial additive produced from Euglena gracilis, providing a rich source of the β‐1,3‐glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune^? and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune^? for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune^? (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune^? kg^?1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune^? had lowest numerical mortality values. Therefore, Algamune^? was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo.     

A study on the influence of selected Indian herbal active principles on enhancing the immune system in Fenneropenaeus indicus against Vibrio harveyi infection

Five herbs such as Acalypha indica, Hygrophila spinosa, Picrorhiza kurooa, Tinospora cordifolia and Zingiber officinale were selected to screen for the in vitro immunostimulant activity against the shrimp pathogen Vibrio harveyi using different organic polar and non-polar solvents. After initial screening, the best extract of the five herbs was mixed thoroughly in equal proportion and made it an artificial immunostimulant diets (ISD) such as ISD-1(500), ISD-2 (1,000), ISD-3 (1,500) and ISD-4 (2,000) mg?kg^?1. The control diet was prepared without adding immunostimulant herbal extracts. The Indian white shrimp, Fenneropenaeus indicus having the weight of 8.0?±?1.0?g, were fed with the above diets for 60?days. After the completion of feeding trail, they were challenged with V. harveyi at the rate of 10⁷CFU/ml and found out the cumulative mortality, growth parameters, haematological and immunological parameters. The weight gain and specific growth rate were significantly (P?<?0.05) increased from the control group to experimental group. The immunostimulant active principles helped to decrease the coagulation time and improved the total haemocyte count (THC), phagocytosis, phenol oxidase (PO) activity, haemagglutinin activity and bacterial clearance.     

Response of the intestinal mucosal barrier of carp (Cyprinus carpio) to a bacterial challenge by Aeromonas hydrophila intubation after feeding with β‐1,3/1,6‐glucan

The effect of dietary β‐glucan on the bacterial community in the gut of common carp (Cyprinus carpio) was examined after oral application of Aeromonas hydrophila. Carp received either feed supplemented with 1% MacroGard^®, a β‐1,3/1,6‐glucan, or a β‐glucan‐free diet. Fourteen days after feeding, half of the carp from each group were intubated with 10⁹ colony‐forming units (CFU) of a pathogenic strain of A. hydrophila. Gut samples were taken 12 hr to 7 days after application and analysed using microbiological and molecular biological techniques (NGS, RT‐PCR‐DGGE). The reaction of the mucosa and the microbiota to an A. hydrophila intubation differed in carp fed with β‐glucan compared to carp from the control group. In β‐glucan fed carp, the total bacterial amount was lower but the number of bacterial species was higher. Bacterial composition was different for carp from both treatment groups. The number of mucin filled goblet cells was reduced in carp fed the β‐glucan diet. Mucus was obviously released from the goblet cells and was probably washed out of the gut together with high numbers of bacteria. This might be protective against pathogenic bacteria and, therefore, feeding with β‐glucan may provide protection against infections of the gut in carp.     

Effects of dietary immunostimulant combination on the growth performance,non‐specific immunity and disease resistance of cobia,Rachycentron canadum (Linnaeus)

The aim of this study was to examine the effects of the immunostimulant combination (IC) containing β‐glucan, A3α‐peptidoglycan, vitamin C and vitamin E on the growth performance, non‐specific immunity and protection against Vibrio harveyi infection in cobia (Rachycentron canadum). Fish were fed diets containing six graded levels of IC (0, 1, 2, 3, 4 and 5 g kg^?1 diet) for 8 weeks. The results showed that the survival rate ranged from 81.1 to 84.4% with no significant difference among all the groups (P > 0.05) after the feeding experiment. Dietary IC significantly increased the specific growth rate (SGR), serum lysozyme, alternative complement pathway (ACH50) activity, phagocytosis percentage (PP) and respiratory burst activity of head kidney macrophages of cobia. Moreover, feeding of supplemented diets containing 3.0 g kg^?1 IC resulted in significantly lower mortality against the pathogens, V. harveyi compared with the control group. To elevate the growth and immune resistance ability of cobia, the optimal dose of dietary IC administration, determined by second‐order polynomial regression analysis was 3.43 and 2.71 g kg^?1 diet, respectively, on the basis of the SGR and mortality after challenge with V. harveyi.     

The feasibility of using primary shrimp hemocyte culture to screen herbal immunostimulants

An analysis of the immune activity of herbal extract in vivo and in vitro assays was performed to explore the feasibility of using primary shrimp hemocyte culture to screen herbal immunostimulants. In vitro experiment was conducted by adding herbal extract into the medium of primary cultured hemocytes and incubating for 24h. In vivo experiment was carried out by feeding shrimp with diets incorporated with herbal extract for 21 days. Challenge test can be used to directly reflect comprehensive immune capacity of organisms. Four immune indexes, total hemocyte counts (THC and lysozyme, phenoloxidase (PO), and phagocytic activity were examined both in vitro and in vivo. The results showed that effects of herbal extract on THC or lysozyme activity were different between in vitro and in vivo. On the contrary, effects of herbal extract on PO activity and phagocytic activity in vitro and in vivo showed a strong correlation. Herbal extract, which enhanced PO activity and phagocytic activity in vitro, had ultimate immune-promoting function in vivo. Therefore, it was feasible to use primary hemocyte culture to screen herbal immunostimulants by determining the effects of herbal immunostimulants on PO and phagocytic activities.     

Effect of oxidized fish oil and α‐tocopherol on growth,antioxidation status,serum immune enzyme activity and resistance to Aeromonas hydrophila challenge of Chinese mitten crab Eriocheir sinensis

The impact of dietary α‐tocopherol on juvenile Chinese mitten crab Eriocheir sinensis was experimentally evaluated in a 10‐week study. Crab were fed with nine diets including three levels of α‐tocopherol (0, 100 and 300 mg kg^?1 diet) and three levels of fish oil oxidation (fresh, moderate and high) in triplicates. Fresh and moderate oil oxidization enhanced weight gain, but moderate and high oil oxidization lowered survival and feed efficiency. The 100‐mg α‐tocopherol kg^?1 diet resulted in higher hepatopancreas MDA than other α‐tocopherol diets. High oil oxidization led to the lowest serum superoxide dismutase (SOD) and glutathione peroxidase (GPH‐PX). The serum SOD and GPH‐PX activities in crab fed 100 mg α‐tocopherol were higher than in those fed other α‐tocopherol diets. The diet without α‐tocopherol addition lowered lysozyme and phenoloxidase (PO) activities compared to other α‐tocopherol diets. Fresh fish oil diet increased PO activity compared to oxidized oils. High oil oxidization caused significantly more mortality than fresh or moderate oxidization after 7‐d postchallenge with Aeromonas hydrophila. Supplementation with α‐tocopherol significantly enhanced resistance to bacterial infection. This study indicates that α‐tocopherol can protect lipid from peroxidation and enhance disease resistance.     

Effects of synbiotic containing Lactobacillus plantarum 7–40 and galactooligosaccharide on the growth performance of white shrimp,Litopenaeus vannamei

This study aimed to develop a synbiotic combination with probiotic, Lactobacillus plantarum 7–40 and one of three prebiotics, fructooligosaccharide (FOS), galactooligosaccharide (GOS) and mannan oligosaccharide (MOS). The best in vitro growth was observed when probiotic was cultured in the medium containing either FOS or GOS as the sole of carbon source. The analysis of enzyme activity revealed that GOS induced the highest activities of protease and β‐galactosidase of probiotic. Based on the findings, probiotic + GOS were selected as synbiotic to evaluate if it could promote the growth of white shrimp, Litopenaeus vannamei. For this, four diets, including a basal diet with no GOS or probiotic (control), 0.4% GOS (PRE), 10⁸ CFU probiotic kg^?1 (PRO) and 0.4% GOS in combination with 10⁸ CFU probiotic kg^?1 (SYN), were fed to shrimp for 60 days, and then the growth performance, intestinal microbiota (including total Vibrio counts, VBCs; and lactic acid bacteria, LAB) and digestive enzyme (including protease, leu‐aminopeptidase and β‐galactosidase) were evaluated. The weight gain (WG) of shrimp fed the PRO did not significantly differ from those of control (p > .05). Shrimp fed the SYN had significantly higher WG compared with the other treatments (p < .05). In addition, the SYN‐fed shrimp had significantly higher LAB and protease, leu‐aminopeptidase and β‐galactosidase activity (p < .05). The lowest presumptive Vibrio count (VBC) was also observed in intestines of SYN‐fed shrimp. Therefore, we suggested that Lac. plantarum 7–40+ GOS can be used as a synergistic synbiotic for shrimp culture.     

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.     

Effects of dietary β‐carotene levels on growth and liver vitamin A concentrations of the soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to evaluate the effect of dietary β‐carotene level on the growth and liver vitamin A concentrations in soft‐shelled turtles, Pelodiscus sinensis, fed a vitamin A‐free diet. Soft‐shelled turtles were fed diets containing 0, 14.5, 26.5, 47.5, 87.3, 112.8 and 163.8 mg β‐carotene kg^?1 for 10 weeks. Although it was not statistically significant due to high deviation within each group, mean weight gain of soft‐shelled turtles fed the diet without β‐carotene supplementation was the lowest among all test groups. Vitamin A concentrations in liver of turtles significantly (P<0.05) increased when dietary β‐carotene level reached 47.5 mg kg^?1 indicating that soft‐shelled turtles were capable of converting β‐carotene to vitamin A. Analysed by regression modelling, dietary β‐carotene levels for optimal growth and maximal liver vitamin A contents of juvenile soft‐shelled turtles fed the vitamin A‐free diets were 49.1 and 88.7 mg kg^?1 respectively.     

Purification and characterization of a β–glucan binding protein from the haemolymph of freshwater prawn Macrobrachium rosenbergii

β‐glucan binding protein (βGBP), a pattern recognition protein was purified from the haemolymph of freshwater prawn Macrobrachium rosenbergii by heparin affinity chromatography that showed a single band in native gradient PAGE. The β‐glucan binding property of the purified protein was confirmed in a phenoloxidase (PO) assay, where addition of βGBP along with β‐glucan increased the specific PO activity compared with that of β‐glucan alone. The molecular weight of the βGBP was found to be ~316 kDa on gel filtration chromatography. In SDS‐PAGE, βGBP molecule was reduced to one polypeptide chain of molecular weight ~113 kDa. Thus the βGBP in M. rosenbergii is possibly a homotrimeric molecule. The purified sample run on unreduced condition in SDS‐PAGE also revealed a similar size band (~113 kDa) and hence, the polypeptide chains of βGBP are held by non‐covalent interactions. The purified βGBP samples run in native PAGE was stained positively with alcian blue for carbohydrates and Sudan black for lipids indicating the βGBP to be a glycolipoprotein. With rabbit polyclonal anti‐βGBP serum developed, an indirect ELISA was standardized and the normal βGBP concentration in adult M. rosenbergii serum was quantified to be ~2 mg mL^?1. Furthermore, the applicability of the developed ELISA is discussed.     

Innate immune response and related gene expression in red swamp crayfish [Procambarus clarkii (Girard)], induced by selenium‐enriched exopolysaccharide produced by bacterium Enterobacter cloacae Z0206

Selenium‐enriched exopolysaccharide (Se‐ECZ‐EPS) produced by bacterium Enterobacter cloacae Z0206 is an important polysaccharide and has been proved its immunostimulating roles in mice. However, relatively less information is available about the potential role of Se‐ECZ‐EPS in crustaceans. To investigate the modulation of immunity in crustaceans, we examined phenoloxidase (PO) activity, respiratory burst (RB) ability, superoxide dismutase (SOD) level and several immune‐related genes' expression profile in red swamp crayfish [Procambarus clarkii (Girard)] that were individually induced with Se‐ECZ‐EPS at 1 μg g^?1. The protection of crayfish against the white spot syndrome virus (WSSV) by Se‐ECZ‐EPS was also investigated. The results showed that PO, RB and SOD activities of crayfish increased significantly (P<0.05) at 24, 48 and 72 h after being injected with Se‐ECZ‐EPS. Among the genes examined, the mRNA expressions of serine proteinase, HSP70 and Mn‐SOD were significantly up‐regulated at 24 and 48 h after the Se‐ECZ‐EPS treatment, while no significant changes in the expression in proPO were observed from 24 to 72 h. The survival rate of crayfish that received Se‐ECZ‐EPS was significantly higher than that of the control group after 14 days (59.7% and 0% respectively). Taken together, it can be concluded that Se‐ECZ‐EPS is an efficient immunostimulant and can improve the immunity of crayfish.