White shrimp Litopenaeus vannamei that received the hot-water extract of Gracilaria tenuistipitata showed earlier recovery in immunity after a Vibrio alginolyticus injection

White shrimp Litopenaeus vannamei which had been immersed in seawater containing the hot-water extract of Gracilaria tenuistipitata at 0 (control), 200, 400, and 600 mg L^?1 for 3 h, were challenged with Vibrio alginolyticus at 4.6 × 10⁶ colony-forming units (CFU) shrimp^?1 and then placed in normal seawater (34‰). The survival rates of shrimp immersed in 200, 400, and 600 mg L^?1 of the hot-water extract were significantly higher than those of control shrimp over 48–120 h. In another experiment, L. vannamei which had been immersed in the hot-water extract at 0, 200, 400, and 600 mg L^?1 for 3 h, were challenged with V. alginolyticus at 4.0 × 10⁶ CFU shrimp^?1, and the immune parameters examined included the haemocyte count, phenoloxidase (PO) activity, respiratory burst (RB), and superoxide dismutase (SOD) activity at 12–120 h post-challenge after shrimp had been released into normal seawater. Shrimp not exposed to the hot-water extract or V. alginolyticus served as the background control. Results indicated that the haemocyte count, PO activity, RB, and SOD activity of shrimp immersed in 600 mg L^?1 were significantly higher than those of control shrimp at 12–72 h post-challenge. Results also indicated that total haemocyte count (THC), PO activity, RB and SOD activity of shrimp immersed in 400 and 600 mg L^?1 of the hot-water extract returned to the background values at 96, 48, 48, and 72 h, whereas these parameters of control shrimp returned to the original values at >120, >120, 96, and 96 h post-challenge, respectively. It was therefore concluded that L. vannamei that had been immersed in seawater containing the hot-water extract of G. tenuistipitata exhibited protection against V. alginolyticus as evidenced by the earlier recovery of immune parameters.     

Administration of the hot-water extract of Spirulina platensis enhanced the immune response of white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus

White shrimp Litopenaeus vannamei which had been injected with the hot-water extract of Spirulina platensis at 6, 10, and 20 μg g^?1, or immersed in aerated seawater containing extract at 200, 400, and 600 mg L^?1 were challenged with Vibrio alginolyticus at 1.5 × 10⁶ or 1.4 × 10⁶ colony-forming units (cfu) shrimp^?1, and then placed in seawater. Survival rates of shrimp that received the extract of S. platensis at 6–20 μg g^?1, and those of shrimp immersed in seawater containing the extract at 400 and 600 mg L^?1 were significantly higher than those of control shrimp after 24–96 and 48–96 h, respectively. In a separate experiment, the hyaline cell (HC) count, granular cell (GC, including semi-granular cell) count, total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, and lysozyme activity were measured when shrimp were injected with the extract at 6, 10, and 20 μg g^?1, and immersed in seawater containing the extract at 200, 400, and 600 mg L^?1. These parameters directly increased with the concentration, and significantly increased when shrimp were immersed in the seawater containing the extract at 0.5–4 h. L. vannamei that received all doses of the extract via injection or via immersion all had increased phagocytic activity and clearance efficiency to V. alginolyticus at 12–72 h and 3–4 h, respectively. It was concluded that L. vannamei that received the hot-water extract of S. platensis had enhanced innate immunity and increased resistance against V. alginolyticus infection.     

Effect of hypoxic stress on the immune response and the resistance to vibriosis of the shrimpPenaeus stylirostris

The immune response of shrimp exposed to severe hypoxia, 1 mg O₂ml⁻¹for 24 h, was measured in terms total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase activity (PO) and respiratory burst by NBT reduction. Results showed that hypoxia induced a significant decrease (P<5%) of the THC due to a significant decrease of semi-granular cells (SGC) (P<5%) and hyaline cells (HC) (P<5%). There was also a decrease in the NBT reduction. On the other hand, PO activity increased significantly (P<1%) and seemed to be related to a lower amount of plasma inhibitors regulating the prophenoloxidase system. Experimental infection with a virulentVibrio alginolyticuswas carried out by intramuscular injection of 2×10⁴colony forming units (cfu) per shrimp. The mortality of stressed shrimp was 48% while the mortality of control shrimp was 32%. This difference was significant (P<1%). The variations of immunological parameters, except for the PO activity, seem to be consistent with thein vivoresponse against the pathogenicVibrio. Coyright 1998 Academic Press     

Fucoidan effectively provokes the innate immunity of white shrimp Litopenaeus vannamei and its resistance against experimental Vibrio alginolyticus infection

In this study, we examined the effect of fucoidan on the immune response of white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus infection. Fucoidan induced degranulation, caused changes in the cell morphology, and increased activation of prophenoloxidase (proPO) and the production of superoxide anions in vitro. Shrimp that received fucoidan via immersion at 100, 200, and 400 mg l^?1 after 3 h showed haemocyte proliferation and a higher mitotic index of haematopoietic tissue. In another experiment, the haemocyte count, phenoloxidase (PO) activity, and respiratory bursts (RBs) were examined after the shrimp had been fed diets containing fucoidan at 0 (control), 0.5, 1.0, and 2.0 g kg^?1 for 7–21 days. Results indicated that these parameters directly increased with time. The immune parameters of shrimp fed the 1.0 g kg^?1 diet were significantly higher than those of shrimp fed the 2.0 g kg^?1 diet after 14 and 21 days. Phagocytic activity and the clearance efficiency against V. alginolyticus were significantly higher in shrimp fed the 1.0 g kg^?1 diet compared to those of shrimp fed the 0, 0.5 and 2.0 g kg^?1 diets. In a separate experiment, shrimp that had been fed diets containing fucoidan for 21 days were challenged with V. alginolyticus at 10⁶ colony-forming units shrimp^?1. Survival rates of shrimp fed the 1.0 and 2.0 g kg^?1 diets were significantly higher than those of shrimp fed the 0 and 0.5 g kg^?1 diets for 96–120 h. We concluded that fucoidan provokes innate immunity of shrimp as evidenced by haemocyte degranulation, proPO activation, and the mitotic index of haematopoietic tissue, and that dietary administration of fucoidan at 1.0 g kg^?1 enhanced the immune response of shrimp and their resistance against V. alginolyticus infection.     

Shrimp that have received carrageenan via immersion and diet exhibit immunocompetence in phagocytosis despite a post-plateau in immune parameters

The effect of carrageenan on the immune response of white shrimp Litopenaeus vannamei, was studied in vitro and in vivo. Shrimp haemocytes receiving carrageenan at 1 mg ml⁻¹ experienced change in cell size, reduction in cell viability, increase in PO activity, serine proteinase activity, and RB in vitro. Shrimp received carrageenan via immersion at 200, 400 and 600 mg L⁻¹ after 3 h and orally at 0.5, 1.0 and 2.0 g kg⁻¹ after 3 weeks showed higher proliferation of haematopoietic tissues (HPTs) together with increases in haemocyte count and other immune parameters. Shrimp that fed a diet containing carrageenan at 0.5 g kg⁻¹ after 3 weeks significantly up-regulated gene expressions of several immune-related proteins. The immune parameters of shrimp that received carrageenan via immersion and orally increased to a plateau after 3 h and after 3 weeks, but decreased after 5 h and 6 weeks, respectively. Phagocytosis and clearance of Vibrio alginolyticus remained high in shrimp that had received carrageenan via immersion after 5 h and orally after 6 weeks, respectively. Resistances of shrimp against V. alginolyticus and white spot syndrome virus were higher over 24–144 h and 72–144 h, respectively in shrimp that received carrageenan at 600 mg L⁻¹ via immersion after 3 and 5 h. It was concluded that carrageenan effectively triggers an innate immunity in vitro, and increases mitotic index of HPT, immune parameters, gene expressions and resistance against pathogens in vivo. Shrimp received carrageenan via immersion and orally exhibited immunocompetence in phagocytosis and clearance of V. alginolyticus, and resistance to pathogen despite the trend in immune parameters to recover to background values.     

Effects of the dietary administration of sodium alginate on the immune responses and disease resistance of Taiwan abalone,Haliotis diversicolor supertexta

Sodium alginate extracted from brown algae was reported to enhance the immune response and resistance of fish and shrimp. In this study, survival rates of the abalone, Haliotis diversicolor supertexta, against Vibrio parahaemolyticus, and its non-specific immune parameters such as the total haemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts, superoxide dismutase (SOD) activity, phagocytic activity, and clearance efficiency to V. parahaemolyticus by H. diversicolor supertexta were determined when abalone (4.5 ± 0.4 g) were fed diets containing sodium alginate at 0, 1.0, 2.0, and 3.0 g kg^?1. Abalone fed a diet containing sodium alginate at 2.0 and 3.0 g kg^?1 for 14 days and at 1.0 g kg^?1 for 21 days had significantly higher survival rates than those fed the control diet after challenge with V. parahaemolyticus. The relative survival percentages of abalone fed the 1.0, 2.0, and 3.0 g kg^?1 sodium alginate-containing diets for 14 and 21 days were 16.1%, 40.0%, and 48.0%, and 63.6%, 27.3% and 22.6%, respectively. The PO activity, respiratory bursts, SOD activity, and phagocytic activity and clearance efficiency of V. parahaemolyticus of abalone fed the sodium alginate-containing diets at 1.0, 2.0, and 3.0 g kg^?1 were significantly higher than those of abalone fed the control diet for 14 days. After 21 days, the PO activity, respiratory bursts, SOD activity, and phagocytic activity and clearance efficiency of V. parahaemolyticus by abalone fed the sodium alginate-containing diet at 1.0 g kg^?1 were significantly higher than those of abalone fed the other diets. It was concluded that sodium alginate can be used as an immunostimulant for abalone through dietary administration to enhance immune responses of abalone and resistance against V. parahaemolyticus, which were related to the dose and timing of administration.     

Modulation of the innate immune system in white shrimp Litopenaeus vannamei following long-term low salinity exposure

Immune parameters, haemocyte lifespan, and gene expressions of lipopolysaccharide and β-glucan-binding protein (LGBP), peroxinectin (PX), integrin β, and α2-macroglobulin (α2-M) were examined in white shrimp Litopenaeus vannamei juveniles (0.48 ± 0.05 g) which had been reared at different salinity levels of 2.5‰, 5‰, 15‰, 25‰, and 35‰ for 24 weeks. All shrimp survived during the first 6 weeks. The survival rate of shrimp reared at 2.5‰ and 5‰ was much lower (30%) than that of shrimp reared at 15‰, 25‰, and 35‰ (76%~86%) after 24 weeks. Shrimp reared at 25% grew faster. Shrimp reared at 2.5‰ and 5‰ showed lower hyaline cells (HCs), granular cells (GCs), phenoloxidase activity (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, and lysozyme activity, but showed a longer haemocyte lifespan, and higher expressions of LGBP, PX, integrin β, and α2-M. In another experiment, shrimp which had been reared at different salinity levels for 24 weeks were challenged with Vibrio alginolyticus (6 × 10(6) cfu shrimp(-1)), and WSSV (10(3) copies shrimp(-1)) and then released to their respective seawater. At 96-144 h, cumulative mortalities of shrimp reared at 2.5‰ and 5‰ were significantly higher than those of shrimp reared at 15‰, 25‰, and 35‰. It was concluded that following long-term exposure to 2.5‰ and 5‰ seawater, white shrimp juveniles exhibited decreased resistance against a pathogen due to reductions in immune parameters. Increases in the haemocyte lifespan and gene expressions of LGBP, integrin β, PX, and α2-M indicated that shrimp had the ability to expend extra energy to modulate the innate immune system to prevent further perturbations at low salinity levels.     

The immune response of white shrimp Litopenaeus vannamei following Vibrio alginolyticus injection

White shrimp Litopenaeus vannamei injected with saline, and injected with tryptic soy broth (TSB)-grown Vibrio alginolyticus at 1.0 x 10(5) and 1.8 x 10(5) colony-forming units (cfu) shrimp(-1) were examined for hyaline cell (HC) counts, granular cell (GC) counts, total haemocyte counts (THCs), phenoloxidase (PO) activity, respiratory burst (RB) and superoxide dismutase (SOD) activity after 1-168 h. Shrimp that received no injection served as the control. The shrimps which received V. alginolyticus at both doses showed significant decreases in these parameters after 6-96 h. The values for HC and SOD activity decreased earlier and then RB. The time to cause maximum depletion of haemocytes (haemocytopenia), PO activity, RB, and SOD activity were 12, 72, 48, and 24 h post-injection, respectively. The HC, GC, and RB returned to the original values earlier at 72 h, followed by SOD activity at 96 h, and then PO activity at 168 h post-infection. It was concluded that an injection of V. alginolyticus rapidly reduced the shrimp's immunity by decreasing HC, GC, SOD activity, RB, and PO activity within 3-24 h, followed by a slow recovery during 72-168 h post-injection. Furthermore, white shrimp L. vannamei which received V. alginolyticus showed a 6-9 h later response in PO activity, and a 72-96 h later recovery of PO activity, compared to the responses in RB and SOD activity indicating their roles in shrimp defence and immunity.     

Noradrenaline modulates the immunity of white shrimp Litopenaeus vannamei

The total haemocyte count (THC), phenoloxidase activity, respiratory burst, superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency in response to pathogen Vibrio alginolyticus were measured when the white shrimp Litopenaeus vannamei (18.4 +/- 1.2 g) were injected individually with noradrenaline at 10(-8), 10(-7) and 10(-6) mol shrimp(-1). For the shrimp that received noradrenaline at 10(-8), 10(-7) and 10(-6) mol shrimp(-1), the THC decreased by 15%, 21% and 32%, phenoloxidase activity decreased by 15%, 31% and 31%, respiratory burst decreased by 13%, 21% and 32%, and SOD activity decreased by 46%, 56% and 55%, respectively, after 2 h. The phagocytic activity and clearance efficiency of shrimp that received noradrenaline at either dose decreased significantly after 2 h. The THC, phenoloxidase activity, respiratory burst, SOD activity, phagocytic activity and clearance efficiency returned to normal values after 4, 4, 8, 24, 16 and 8 h, respectively, in the shrimp that received noradrenaline at either dose. In another experiment, L. vannamei which had received noradrenaline at 10(-8), 10(-7) and 10(-6) mol shrimp(-1) were challenged after 1h by injection with V. alginolyticus at 1.0 x 10(5) colony-forming units (cfu)shrimp(-1) and then placed in seawater of 20 per thousand. The cumulative mortality of shrimp that received noradrenaline at either dose was significantly higher than that of shrimp that received saline after 4 h, and at the termination of the experiment (48 h after the challenge). It is therefore concluded that noradrenaline administration at 10(-6) mol shrimp(-1) or less causes immune modulation of L. vannamei.     

Effects of dopamine on the immunity of white shrimp Litopenaeus vannamei

The total haemocyte count (THC), phenoloxidase activity, respiratory burst, superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency in response to pathogen Vibrio alginolyticus were measured when the white shrimp Litopenaeus vannamei (20.0+/-1.5 g) were injected individually with dopamine at 10(-8), 10(-7) and 10(-6)mol shrimp(-1), respectively. For the shrimp that received dopamine at 10(-7) and 10(-6)mol shrimp(-1), the THC decreased by 25% and 39%, phenoloxidase activity decreased by 15% and 32%, respiratory burst decreased by 21% and 36%, and SOD activity decreased by 50% and 63%, respectively, after 4 h. The phagocytic activity and clearance efficiency of shrimp that received dopamine at either dose decreased significantly after 2 h. The THC, phenoloxidase activity, respiratory burst, SOD activity, phagocytic activity and clearance efficiency returned to normal values after 16, 8, 8, 24, 16 and 4 h, respectively, for the shrimp that received dopamine at either dose. In another experiment, L. vannamei which had received dopamine at 10(-8), 10(-7) and 10(-6)mol shrimp(-1) were challenged after 1 h by injection with V. alginolyticus at 1.0x10(5) colony-forming units (cfu) shrimp-1 and then placed in seawater of 20 per thousand. The cumulative mortality of shrimp that received dopamine at either dose was significantly higher than that of shrimp that received saline after 8 h, and of shrimp that received saline at the termination of the experiment (48 h after the challenge). It is therefore concluded that dopamine administration at 10(-6)mol shrimp-1 or less causes immune modulation of L. vannamei.     

An immersion of Gracilaria tenuistipitata extract improves the immunity and survival of white shrimp Litopenaeus vannamei challenged with white spot syndrome virus

The innate immunity and resistance against white spot syndrome virus (WSSV) in white shrimp Litopenaeus vannamei which received the Gracilaria tenuistipitata extract were examined. Shrimp immersed in seawater containing the extract at 0 (control), 400 and 600 mg L(-1) for 3 h were challenged with WSSV at 2 × 10(4) copies shrimp(-1). Shrimp not exposed to the extract and not received WSSV challenge served as unchallenged control. The survival rate of shrimp immersed in 400 mg L(-1) or 600 mg L(-1) extract was significantly higher than that of challenged control shrimp over 24-120 h. The haemocyte count, phenoloxidase activity, respiratory burst, superoxide dismutase activity, and lysozyme activity of shrimp immersed in 600 mg L(-1) extract were significantly higher than those of unchallenged control shrimp at 6, 6, 6, 6, and 6-24 h post-challenge. In another experiment, shrimp which had received 3 h immersion of 0, 400, 600 mg L(-1) extract were challenged with WSSV. The shrimp were then received a booster (3 h immersion in the same dose of the extract), and the immune parameters were examined at 12-120 h post-challenge. The immune parameters of shrimp immersed in 600 mg L(-1) extract, and then received a booster at 9, 21, and 45 h were significantly higher than those of unchallenged control shrimp at 12-48 h post-challenge. In conclusion, shrimp which had received the extract exhibited protection against WSSV as evidenced by the higher survival rate and higher values of immune parameters. Shrimp which had received the extract and infected by WSSV showed improved immunity when they received a booster at 9, 21, and 45 h post-WSSV challenge. The extract treatment caused less decrease in PO activity, and showed better performance of lysozyme activity and antioxidant response in WSSV-infected shrimp.     

Generation of superoxide anion and SOD activity in haemocytes and muscle of American white shrimp (Litopenaeus vannamei) as a response to beta-glucan and sulphated polysaccharide

Juvenile American white shrimp (Litopenaeus vannamei) were immersed in aerated beta-glucan and sulphated polysaccharide solutions for 1, 3 and 6 h. Superoxide anion and SOD activity in haemocytes and muscle were investigated to evaluate whether beta-glucan and sulphated polysaccharide induce any immunostimulatory activity. Haemocytes and muscle showed different levels of superoxide anion generation and SOD activity (2.0 and 14 times that of control, respectively) when shrimp were immersed for 6 h in aerated sea water containing beta-glucan and sulphated polysaccharide. Total haemocyte count (THC) decreased within the first 24 h after challenge with immunostimulants, but THC and total soluble haemocyte protein increased over normal values after 48-120 h. Single immunostimulation with beta-glucan and sulphated polysaccharide is capable of generating an increase in the respiratory burst of L. vannamei haemocytes.     

Immune responses of Litopenaeus vannamei to thermal stress: a comparative study of shrimp in freshwater and seawater conditions

The effects of hypothermal (22–16?°C) and hyperthermal (22–28?°C) stress on the immune system responses of the shrimp Litopenaeus vannamei cultured in either freshwater or seawater were measured and compared. The following immune system indicators were measured for comparison: total hemocyte count (THC), activity of phenoloxidase (PO), nitric oxide synthase (NOS), superoxidase (SOD), and malondialdehyde (MDA) content. Thermal stress significantly decreased THC in both freshwater and seawater shrimp within 6–12?h (P?0.05). After hypothermal stress, all shrimp had a significantly lower THC level than their prechallenge levels (P?0.05). Under both types of thermal stress, the activity of PO, NOS, and SOD first increased and then decreased. After 48?h of thermal stress, shrimp PO and NOS activity decreased in both freshwater and seawater. After 48?h of thermal stress, the reduction in the SOD activity in the hemolymph of freshwater shrimp was greater than that in seawater shrimp. During exposure to stress, the MDA content in freshwater shrimp was significantly higher than in seawater shrimp, which demonstrated that lipids in freshwater shrimp were more susceptible to peroxidation than those in seawater shrimp, particularly at low temperatures. Large temperature fluctuations, particularly sudden cooling, should be avoided when rearing L. vannamei because of high rates of lipid peroxidation and decreased immunity. These effects are more marked in freshwater than in seawater.     

Effect of saponin immersion on enhancement of the immune response of white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus

The haemocyte count, phenoloxidase (PO) activity, specific alpha(2)-macroglobulin (alpha2-M) activity, respiratory burst, superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, phagocytic activity, and clearance efficiency against Vibrio alginolyticus were examined when the white shrimp Litopenaeus vannamei (10.42+/-2.0g) were immersed in seawater (34 per thousand) containing different concentrations of saponin (0, 0.5, 1 and 2mgL(-1)) for 24, 48 and 72h. Hyaline cells (HC), the total haemocyte count (THC), specific alpha2-M activity, respiratory burst, SOD activity, and GPx activity directly increased with the saponin concentration, whereas PO activity was inversely related to the saponin concentration. White shrimp L. vannamei that were immersed in saponin at 1 and 2mgL(-1) showed increased phagocytic activity and clearance efficiency to V. alginolyticus over 48-72h. In another experiment, shrimp immersed in seawater containing different concentrations of saponin after 72h were challenged with V. alginolyticus at 3.2x10(5) colony-forming units (cfu)shrimp(-1), and then placed in seawater. The survival rate of shrimp immersed in seawater containing saponin at either dose was significantly higher than that of control shrimp after 12h, as well as at the termination of the experiment (5days after the challenge). It was therefore concluded that L. vannamei immersed in water containing saponin at 2mgL(-1) or less exhibited an immunomodulatory effect, as well as protection against V. alginolyticus infection.     

Enhancement of immunity and disease resistance in the white shrimp,Litopenaeus vannamei,by the probiotic,Bacillus subtilis E20

Effects of Bacillus subtilis E20 isolated from fermented soybean on immune parameters and the disease resistance of the white shrimp (Litopenaeus vannamei) after 98 days of B. subtilis E20 feeding were evaluated in this study. Shrimp fed B. subtilis E20-containing diets at concentrations of 10⁶ (E206), 10⁷ (E207), and 10⁸ (E208) cfu kg^?1, respectively, had significantly increased survival rates of 13.3%, 16.7%, and 20%, compared to the control (fed no probiotic) after being challenged with Vibrio alginolyticus. There were no significant differences in the total hemocyte count, respiratory burst, or superoxide dismutase glutathione peroxidase among all treatments. Shrimp fed a higher concentration of the probiotic (E208) exhibited significant increases in phenoloxidase activity, phagocytic activity, and clearance efficiency compared to control shrimp. In addition, B. subtilis E20 showed a weaker inhibitory effect against the growth of Aeromona hydrophila with around a 0.3-cm inhibitory zone, but showed no inhibitory effects against other selected pathogens, such as white shrimp pathogens: V. alginolyticus and Vibrio vulnificus. These results suggest that the increased resistance of shrimp after B. subtilis E20 consumption occurs through immune modifications, such as increases in phenoloxidase activity, phagocytic activity, and clearance efficiency against V. alginolyticus.     

The effect of different acclimation temperatures on the prophenoloxidase system and other defence parameters in Litopenaeus vannamei

Water temperature changes (higher and lower than 24 degrees C) were shown to have a significant effect on dopamine (DA) concentration, haemocyte count and the proPO system in the white shrimp Litopenaeus vannamei. No significant difference in any of the parameters was observed in the control group. DA concentration in haemolymph in the experimental groups increased to a peak value at 0.5 days; meanwhile serine protease (SP) activity and proteinase inhibitor (PI) activity decreased. Total haemocyte count (THC), differential haemocyte count (DHC) and PO activity were lowest at 1 day. All defence parameters became stable after 1-3 days, while the total haemocyte and large granular cell count stabilized after 6 days. After these stabilized, there was no significant difference in DA concentration and PI activity between the control and experimental groups, as was the case for the THC, DHC, PO and SP activities of shrimp held at higher temperatures. However these latter four parameters in the lower temperature groups were distinctly lower than the control group. alpha(2)-Macroglobulin activity in the experimental groups increased to a peak value after 1 day compared with the control and then stabilized after 6 days when the activity levels in higher temperature groups were higher than the control, while the lower temperature groups had no significant difference from the control. It was therefore concluded that water temperature changes modulated the immune system of L. vannamei.     

Immune responses of Fenneropenaeus chinensis against white spot syndrome virus after oral delivery of VP28 using Bacillus subtilis as vehicles

The protective efficacy of oral administration of VP28 using Bacillus subtilis as vehicles (rVP28-bs) in shrimp, Fenneropenaeus chinensis, upon challenge with white spot syndrome virus (WSSV) was investigated. The calculated relative percent survival (RPS) value of rVP28-bs fed shrimp was 83.3% when challenged on the 14th day post-administration, which is significantly higher (p < 0.001) than that of the group administered recombinant Escherichia coli over-expressing rVP28 (rVP28-e21). After immunization, activities of phenoloxidase (PO), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) in hemolymph were analyzed. It was found that the supplementation of rVP28-bs into shrimp food pellets resulted in the most pronounced increase of iNOS activity (p < 0.001), but had the least influence on activities of PO and SOD. Besides, in the shrimp orally administered with rVP28-bs, the caspase-3 activity was one-fifth that of the control, though the signs of apoptosis (chromatin margination, nuclear fragmentation and apoptotic bodies) could not be observed by transmission electron microscope (TEM). These results suggest that by oral delivery of rVP28-bs, shrimp showed significant resistance to WSSV and an effect on the innate immune system of shrimp. The remarkably enhanced level of iNOS after rVP28-bs administration might be responsible for antiviral defense in shrimp.