Essential oil of Lippia alba in the diet of Macrobrachium rosenbergii: Effects on antioxidant enzymes and growth parameters

The juveniles of Macrobrachium rosenbergii (0.060 ± 0.004 g and 2.018 ± 0.071 cm) were fed during 60 days with diets containing different doses of Lippia alba essential oil (EOLA) (0.0—control, 1.0, or 2.0 ml EOLA/kg diet; in triplicate, with 20 prawns/replicate). After the experimental period, were verified the survival, growth parameters and the antioxidant enzymes, lipid peroxidation and Na/K‐ATPase activities in hepatopancreas and gills, respectively, of the animals. There were no significant differences on survival, growth parameters and gill Na/K‐ATPase activity. However, glutathione peroxidase and glutathione‐S‐transferase activities were lower in hepatopancreas of prawns fed with 1.0 ml EOLA/kg diet compared to the control. Moreover, the activity of the enzyme superoxide dismutase was higher in hepatopancreas of prawns that received 2.0 ml EOLA/kg diet compared to those fed with 1.0 ml EOLA/kg diet. Although the addition of both EOLA doses has not improved the survival and growth parameters of M. rosenbergii, these doses contributed to decrease lipid peroxidation. Additionally, the dose of 2.0 ml EOLA/kg diet contributed for increasing the antioxidant enzymes activities in the hepatopancreas, improving antioxidant status, and therefore, it can be recommended as diet supplementation for M. rosenbergii.     

Methionine hydroxy analogue prevents oxidative damage and improves antioxidant status of intestine and hepatopancreas for juvenile Jian carp (Cyprinus carpio var. Jian)

Oxidative damage and antioxidant status of intestine and hepatopancreas for juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of methionine hydroxy analogue (MHA: 0, 5.1, 7.6, 10.2, 12.7, 15.3 g kg^?1 diet) for 60 days were studied. Radical scavenging ability, antioxidant enzymes activities such as superoxide dismutase (SOD), catalase (CAT), glutathione‐S‐transferase (GST), glutathione peroxidase (GPX) and glutathione reducase (GR), as well as glutathione (GSH), protein carbonyl (PC) and malondialdehyde (MDA) contents were assayed in these tissues. Results indicated that anti‐superoxide anion capacity in intestine and anti‐hydroxyl radical capacity in hepatopancreas significantly improved with dietary MHA levels up to 7.6 and 10.2 g kg^?1 diet respectively, whereupon they decreased (P < 0.05). SOD, CAT, GST, GPX, GR activities in intestine and hepatopancreas, as well as GSH content in hepatopancreas significantly increased with optimal MHA levels which were in the range of 5.1–10.2 g kg^?1 diet, and thereafter decreased (P < 0.05). Meanwhile, MDA and PC contents in these tissues together with GOT and GPT activities in plasma significantly decreased with optimal MHA levels which were in the range of 5.1–7.6 g kg^?1 diet, and thereafter increased (P < 0.05). These results suggested that MHA improved antioxidant status and depressed lipid and protein oxidation in intestine and hepatopancreas.     

The antioxidant capacity response to hypoxia stress during transportation of characins (Hyphessobrycon callistus Boulenger) fed diets supplemented with carotenoids

This study aimed to determine whether dietary carotenoid (CD) supplements could affect the antioxidant capacity of characins Hyphessobrycon callistus upon hypoxia stress at live transportation. Two types of CD [astaxanthin (AX), β‐carotene (BC)] and their 1:1 combination (MX) at three concentrations (10, 20 and 40 mg kg^?1) were supplemented, resulting in nine CD diets. After 8 weeks' rearing, the resulting fish were divided into two subgroups and exposed to hypoxia or normoxia. Hypoxia involved a gradual decrease in dissolved oxygen (DO) from 6.5 to <1.0 mg L^?1. Normoxia was DO kept in saturation. Hypoxia led to an increase in the total antioxidant status (TAS), superoxide dismutase (SOD), glutathione peroxidases (GPx) and aspartate aminotransferase (AST) activity of blood serum in fish, but had no effect on alanine aminotransferase (ALT). Under hypoxia, fish fed CD diets had lower SOD, GPx and ALT activity than control fish, showing that dietary CD could increase the antioxidant capacity and protection of the liver. Dietary AX was more effective for antioxidant capacity than BC and MX when under hypoxia stress, because GPx, ALT and AST were lower in AX‐fed fish. Except TAS, the other four enzyme activities showed decreasing trends with increasing dietary CD concentrations.     

Dietary phosphorus prevents oxidative damage and increases antioxidant enzyme activities in intestine and hepatopancreas of juvenile Jian carp

This study was conducted to investigate the effect of dietary phosphorus on the intestine and hepatopancreas antioxidant capacity of juvenile Jian carp (Cyprinus carpio var. Jian). Jian carp, with an average initial weight of 7.17 ± 0.01 g, were fed with diets containing graded concentrations of available phosphorus, namely 1.7 (control), 3.6, 5.5, 7.3, 9.2 and 11.0 g kg^?1 diet for 9 weeks. Results showed that, in intestine and hepatopancreas, content of malondialdehyde (MDA), protein carbonyl (PC) and glutathione (GSH), capacity of anti‐superoxide anion (ASA) and anti‐hydroxyl radical (AHR), and glutathione reductase (GR), catalase (CAT), glutathione S‐transferase (GST), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were significantly affected by dietary phosphorus levels (P < 0.05). Regression analysis showed that significant quadratic responses occurred in MDA content and ASA, GST, GPx and AHR activities in intestine, GSH content and CAT and SOD activities in hepatopancreas (P < 0.05). These results indicate that optimal level of dietary phosphorus prevented oxidative damage and increased antioxidant enzyme activities in the intestine and hepatopancreas of juvenile Jian carp. The phosphorus requirement estimated from MDA using quadratic regression analysis was 5.7 g kg^?1 diet.     

Effects of dietary vitamin K levels on growth performance,enzyme activities and antioxidant status in the hepatopancreas and intestine of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1400 juvenile Jian carp (Cyprinus carpio var. Jian) (8.93 ± 0.03 g) were fed diets containing graded levels of vitamin K at 0.027 (basal diet), 1.52, 3.02, 4.51, 6.02 and 7.52 mg kg^?1 diet for 60 days to investigate the effects of vitamin K on growth, enzyme activities and antioxidant capacity in the hepatopancreas and intestine. Percentage weight gain (PWG), feed intake and feed efficiency of fish were improved by vitamin K. Activities of trypsin, chymotrypsin, amylase and lipase in the intestine and hepatopancreas and Na⁺, K⁺‐ATPase, creatine kinase, alkaline phosphatase and gamma‐glutamyl transpeptidase in the intestine were increased by vitamin K. Malondialdehyde and protein carbonyl contents in the hepatopancreas and intestine were decreased with vitamin K supplements. Certain level of vitamin K increased antihydroxyl radical, antisuperoxide anion, superoxide dismutase, catalase, glutathione‐S‐transferase, glutathione peroxidase and glutathione reductase activities and glutathione contents in the hepatopancreas and intestine. Intestinal Lactobacillus, E. coli and Aeromonas were changed with vitamin K supplements. Together, these results indicate that vitamin K improved fish growth, digestive and absorptive ability, and anti‐oxidant capacity. The dietary vitamin K requirement of juvenile Jian carp (8.93–73.7 g) based on PWG was 3.13 mg kg^?1 diet.     

Dietary astaxanthin improved the body pigmentation and antioxidant function,but not the growth of discus fish (Symphysodon spp.)

To assess the effects of dietary astaxanthin on the growth and body colour of red discus fish (Symphysodon spp.), synthetic astaxanthin was added into the basal diet (beef heart hamburger) with the levels of 0 (control diet), 50, 100, 200, 300 and 400 mg kg^?1 respectively. The six experimental diets were fed to discus fish with an initial body weight of 10.3 ± 0.8 g for 8 weeks. The results showed that the supplementation of 50–200 mg kg^?1 astaxanthin had no significant effects on growth performance of discus fish, but the high supplementation of astaxanthin (300 or 400 mg kg^?1) significantly reduced the weight gain and increased the feed coefficient ratio (P < 0.05). After 4 or 8 weeks of feeding, the L* (lightness) values in astaxanthin‐supplemented groups were significantly lower, while a* (redness), b* (yellowness) and skin astaxanthin contents were significantly higher than the control group (P < 0.05). When the astaxanthin supplementation reached 200 mg kg^?1, skin redness and astaxanthin contents remained relatively stable. When b* was relatively stable, the supplemental astaxanthin was 300 (4 weeks) and 50 mg kg^?1 (8 weeks) respectively. With the supplemental astaxanthin increasing, the astaxanthin retention rate significantly decreased and hepatic total antioxidant capacity was strengthened. The dietary astaxanthin also significantly increased the reduced glutathione level (P < 0.05) when the astaxanthin inclusion was higher than 50 mg kg^?1. The above results showed that dietary astaxanthin could effectively improve the skin pigmentation of red discus fish in 4 weeks and the supplementation level was suggested to be 200 mg kg^?1.     

Histopathological changes in giant freshwater prawn Macrobrachium rosenbergii (de Man 1879) fed with probiotic Bacillus licheniformis upon challenge with Vibrio alginolyticus

The effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the histopathological changes in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g) challenged with known pathogenic strain of Vibrio alginolyticus are reported. Two isocaloric basal diets supplemented with probiotic bacteria B. licheniformis (1.0 × 10⁹ cfu/g feed) and other without probiotic supplementation were fed to the M. rosenbergii juveniles for 45 days. The histological observations revealed no significant changes in the hepatopancreas and gut tissues of both the experimental and the control groups which indicate that the present bacterium is a safe candidate probiont for the host. Prawns were challenged with V. alginolyticus after 45 days of feeding with probiotic diet. The histopathological studies of the hepatopancreas revealed that M. rosenbergii fed with probiotic‐supplemented diet showed less changes as compared to the prawns fed with control diet on second and fourth day of post‐experimental challenge with V. alginolyticus. Histopathological observations revealed that the gills of the prawns fed with control diet were severely affected in comparison to the prawns fed with probiotic‐supplemented diet after challenging with V. alginolyticus. Results from this study revealed the improved protection by dietary incorporation of B. licheniformis in reducing the histopathological manifestations due to V. alginolyticus infection in freshwater prawn.     

Effects of dietary astaxanthin on the growth,innate immunity and antioxidant defence system of Paramisgurnus dabryanus

The effects of astaxanthin supplementationon the growth, innate immunity and antioxidant defence system of loach (Paramisgurnus dabryanus) were investigated in this study. A total of 450 fish (initial average weights of 3.00 ± 0.10 g) were fed five diets with graded levels of astaxanthin (0.00, 50.00, 100.00, 150.00 and 200.00 mg/kg) for 56 days. The results showed that astaxanthin supplementation significantly stimulated the growth (FBW, WG, FER, HSI), innate immunity (AKP activity in the hepatopancreas, intestinal tract, muscle and skin and LZM activity in the hepatopancreas, intestinal tract and skin), antioxidant ability (T‐SOD, GSH‐PX and CAT activities and T‐AOC, GSH and MDA contents in the hepatopancreas, intestinal tract, muscle and skin) of loach. Moreover, dietary astaxanthin supplementation significantly up‐regulated the relative mRNA levels of Nrf2 and Maf, and down‐regulated the relative mRNA level of Keap1 in the hepatopancreas when the supplementation levels of astaxanthin were 50–200 mg/kg. In conclusion, the diets with 100–151.06 mg/kg astaxanthin supplementation were optimal for loach, which based on the growth, immunity and antioxidant‐related indicators, and the astaxanthin supplementation regulated the antioxidant ability partially referring to Keap1‐Nrf2 signallings.     

Effects of cholesterol on growth,feed utilization,body composition and immune parameters in juvenile oriental river prawn,Macrobrachium nipponense (De Haan)

An 8‐week feeding trial was conducted to investigate the effects of dietary cholesterol levels on growth, feed utilization, body composition and immune parameters in juvenile oriental river prawn, Macrobrachium nipponense. Six isolipid (80 g kg^?1 crude lipid) and isoproteic (400 g kg^?1 crude protein) diets, supplemented with 0, 3.0, 6.0, 9.0, 12.0 and 15.0 g kg^?1 cholesterol, were evaluated. Growth performance and feed utilization of M. nipponense were improved as dietary cholesterol levels increased. Weight gain and specific growth rate were highest, and feed conversation ratio was lowest, when prawns were fed a diet supplemented with 9.0 g kg^?1 cholesterol. However, final body weights and survival rates of juvenile M. nipponense were not affected significantly by dietary cholesterol. Body composition of prawns, including moisture, crude protein and crude lipid, was not significantly affected by changes in dietary cholesterol. The immune parameters measured in hepatopancreas, including total antioxidant capacity, and glutathione, catalase, alkaline phosphatase and acid phosphatase activities, were at optimum levels in prawns fed with 9.0 g kg^?1 dietary cholesterol. In summary, the best growth performance, lowest feed conversation ratio, and the most enhanced antioxidant capacity and immunity parameters were attained in juvenile M. nipponense when fed a diet supplemented with 9.0 g kg^?1 cholesterol.     

Effects of dietary antioxidants on oxidative stress,antioxidant defence and growth of freshwater crayfish Astacus leptodactylus (Eschscholtz, 1823) during the reproductive period in females

In this study, we examined the effects of dietary antioxidant vitamins [vitamin E (Vit E), vitamin C (Vit C) and vitamin A (Vit A)], astaxanthin (AX) and β‐carotene (βC) supplementation on oxidative stress [malondialdehyde (MDA)], antioxidant enzyme activities [superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH‐Px)] and glutathione (GSH) levels in hepatopancreas, ovarian, muscle, gill tissues and survival rates (SR), live weight gain (LWG), specific growth rate (SGR) of Astacus leptodactylus during ovarian development. The EE, EC, EA, EAX and EβC groups were formed by addingClick here to enter text. Vit E, Vit C, Vit A, AX and βC to the diet respectively. The antioxidants did not have a significant impact on the values of SR, LWG and SGR. After 137 days, the MDA levels in the Control (C) group were statistically higher in the hepatopancreas, ovarian and muscle tissues according negative control (NC), EE, EC, EA, EAX and EβC groups. The SOD activities in ovarian, muscle and gill tissues and GSH levels in ovarian and muscle tissues were lower in the C group compared with the NC group, but GSH‐Px activities in hepatopancreas and ovarian tissues and GSH levels in gill tissues were higher. The MDA levels were statistically lower in all tissues of crayfish in experimental groups according to C group. The antioxidant enzyme activities and GSH levels changed according to tissue. The results showed that Vit E and AX should be supplemented in diets to increase the reproductive performance of crayfish, because Vit E and AX were of higher potential as antioxidants than other vitamins and carotenoids in ovarian tissues.     

Effects of dietary phenylalanine on growth,digestive and brush border enzyme activities and antioxidant capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idella)

This study investigated the effects of phenylalanine on growth, digestive and absorptive ability and antioxidant status of young grass carp (Ctenopharyngodon idella). Young grass carp were fed diets containing 3.4 (basal diet), 6.1, 9.1, 11.5, 14.0 and 16.8 g phenylalanine kg^?1 diet with a fixed of 10.7 g tyrosine kg^?1 diet for 8 weeks. Percent weight gain (PWG), feed efficiency and feed intake of fish were the lowest in fish fed the basal diet (P < 0.05). Trypsin, lipase and amylase activities in the hepatopancreas, and antioxidants including glutathione contents and glutathione reducase activities in the hepatopancreas and intestine were all the highest in fish fed 11.5 g phenylalanine kg^?1 diet (P < 0.05). Trypsin, chymotrypsin and amylase activities in whole intestine, and creatine kinase, Na⁺, K⁺‐ATPase and alkaline phosphatase activities in the proximal intestine, and superoxide dismutase activities in the hepatopancreas and intestine were all the highest when phenylalanine at level of 9.1 g kg^?1 diet (P < 0.05). In conclusion, phenylalanine improved growth, digestive and absorptive ability, and antioxidant capacity of young grass carp. The phenylalanine requirement of young grass carp (256–629 g) based on PWG was 10.4 g kg^?1 diet or 3.44 g 100 g^?1 protein.     

Simplified processing method of banana (Musa acuminata) peels possess the improvement in immunological responses of Macrobrachium rosenbergii

Musa acuminate peel extract as an immunostimulant, administrated in the diets of Macrobrachium rosenbergii, and an assessment of a simplified process to develop low‐cost feed additives are conducted in this study. The products obtained serially, during the processes of hot‐water banana peel extraction, were administrated in the diets of M. rosenbergii, including dried banana peel powder (DBP) at 8 g/kg, hot‐water‐treated banana peel (HBP) at 8 g/kg and hot‐water extract of banana peel (BPE) at 2 g/kg during the 32 days of the feeding trial. Total haemocyte count, different haemocyte count, respiratory bursts; and activity of superoxide dismutase, glutathione peroxidase, phenoloxidase and transglutaminase, as well as an accelerated haemolymph clotting time, significantly increased in M. rosenbergii fed with diets containing HBP and BPE, 32 days post‐feeding. Furthermore, the phagocytic activity and clearance efficiency of prawns against Lactococcus garvieae infection increased significantly. The respective relative survival percentages of prawns fed with HBP, and BPE containing diets after 32 days of feeding trial were 29.2% and 41.7% against L. garvieae infection for 144 hours, and 50%, and 50% against hypothermal stress for 96 hours of exposure. We may therefore conclude that HBP, obtained from a simplified procedure, without centrifugation and lyophilization, may strategically promote tolerance to hypothermal stress, and enhance immunity and resistance to L. garvieae infection.     

Multiple dietary administrating strategies of water hyacinth (Eichhornia crassipes) on enhancing the immune responses and disease resistance of giant freshwater prawn,Macrobrachium rosenbergii

The different products of Eichhornia crassipes leaves including dried E. crassipes powder (DEP), hot‐water treated E. crassipe (HTE), hot‐water extract of E. crassipe (ECE) and dreg of hot‐water extract of E. crassipe extract (dECE) were produced and incorporated into the diet of the prawn, Macrobrachium rosenbergii, as an immunostimulant. Results showed that prawn fed the HTE‐, ECE‐ and dECE‐containing diets for 4 months had increased total haemocyte count, different haemocyte count, phenoloxidase activity, respiratory bursts, superoxide dismutase activity, glutathione peroxidase activity especially in HTE and ECE treatments. The phagocytic activity and clearance efficiency against Lactococcus garvieae of prawn fed the HTE‐ and ECE‐containing diets were significantly higher than those of prawn fed the control diet at 2–4 months. The relative percentage survival of prawn fed the DEP‐, HTE‐, ECE‐ and dECE‐containing diets for 4 months following 144 h challenging with L. garvieae were 19.0%, 38.1%, 38.1% and 33.3%. It was concluded that E. crassipes leaves containing an active component which was easily extracted by hot water can enhance innate immunity and resistance against pathogen of M. rosenbergii by dietary long‐term administration, and the administration of HTE in the diet was the best strategy due to the availability and convenience.     

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.     

Antioxidant defence to ammonia stress of characins (Hyphessobrycon eques Steindachner) fed diets supplemented with carotenoids

This study aimed to find out whether dietary carotenoid (CD) supplement could influence the resistance of characins (Hyphessobrycon eques Steindachner) to ammonia stress. Two types of CD and its combination [astaxanthin (AX), β‐carotene (BC), 1 : 1 combination of AX and BC (MX)] at three concentrations (10, 20 and 40 mg kg⁻¹) were used resulting in nine pigmented diets. No differences in growth and survival of the fish among treatments were found after 8‐week rearing. Experimental and control fish were then exposed to 15 mg total ammonia nitrogen L⁻¹ (stress group) and 0.15 mg total ammonia nitrogen L⁻¹ (normal group) for 72 h, and their blood was withdrawn. No mortality resulted under such TAN concentrations. Serum total antioxidant status (TAS), serum antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidases (GPx)] and serum transaminases [alanine aminotransferase (ALT), aspartate aminotransferase (AST)] were chosen as indices of fish antioxidant capacity or stress resistance. SOD, GPx and AST were affected by the interactions of dietary CD and ammonia stress. The activities of TAS, SOD, GPx and AST increased under the stress. Dietary CD reduced serum SOD, GPx, ALT and AST activities. In conclusion, dietary CD increased the resistance of characins to ammonia stress.     

Identification and characterization of lactic acid bacteria isolated from rainbow trout (Oncorhynchus mykiss,Walbaum 1792), with inhibitory activity against Vagococcus salmoninarum and Lactococcus garvieae

In this study, a total of 98 lactic acid bacteria isolated from rainbow trout intestines were screened for their probiotic properties. The isolates were tested for their ability to inhibit growth of Vagococcus salmoninarum and Lactococcus garvieae. Based on in vitro antagonism, 10 isolates were selected and evaluated pathogenicity in rainbow trout. Isolates were further investigated for hydrophobicity, bile salts and acid tolerance. These isolates were able to survive low pH and high bile concentrations and showed good adherence characteristics. Isolates were characterized phenotypically, and then, 16S rRNA gene sequence analysis was used for confirmation. Selected strains were administered orally at 10⁸ cfu/g feed, and fish were challenged with V. salmoninarum and L. garvieae. The fish fed with lactic acid bacteria supplemented diets did not improve protection against V. salmoninarum. However, administration of Lactococcus lactis subsp. lactis M17 2‐2 and Lactobacillus sakei 2‐3 resulted in a significant reduction in mortality due to L. garvieae when compared to the control fish. RPS values were calculated as 80 and 53% in fish fed with L. sakei 2‐3 and L. lactis subsp. lactis M17 2‐2, respectively. Our results suggest that these strains could provide an alternative for lactococcosis control in aquaculture.     

Effects of dietary supplementation of Haematococcus pluvialis powder on gonadal development,coloration and antioxidant capacity of adult male Chinese mitten crab (Eriocheir sinensis)

The green algae Haematococcus pluvialis is an important source of natural astaxanthin as feed additive. This study was conducted to investigate the effects of dietary supplementation of H. pluvialis powder on gonadal development, coloration and antioxidant capacity of adult male Chinese mitten crab Eriocheir sinensis, and four experimental diets were formulated to contain 0, 0.2%, 0.4% and 0.6% of H. pluvialis powder. There were four treatments (defined as D1~D4) in this study and each treatment had three replicates. Dietary H. pluvialis contents had no significant effects on survival, body weight gain rate and gonadal development of male E. sinensis. For colour parameters, the total carotenoids content in carapace and hepatopancreas as well as hepatopancreatic lightness (L*) and carapace redness (a*) increased significantly with increasing dietary H. pluvialis (p < 0.05). For the antioxidant indices in the serum, D4 had the lowest activities of superoxide dismutase (SOD) and peroxidase (POD), but the highest glutathione peroxidase (GSH‐Px) and lactic dehydrogenase (LDH), while the malondialdehyde (MDA) in serum and hepatopancreas decreased significantly with the rising content of dietary H. pluvialis (p < 0.05); D1 had the highest levels of SOD, POD and GSH‐Px in hepatopancreas. For the non‐specific immune indices, the highest activities of acid phosphatase (ACP) and γ‐glutamyl transpeptidase (γ‐GT) were found on the serum of D3 and D4 (p < 0.05). D1 had the highest levels of ACP and alkaline phosphatase (ALP) in hepatopancreas, while D2 and D3 had the lowest levels of ALP and ACP respectively. These results suggested the optimal dietary natural astaxanthin level was around 40 mg/kg diets.     

Immunomodulatory effect of Cynodon dactylon against white tail disease of giant freshwater prawn,Macrobrachium rosenbergii (de Man, 1879)

The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important and extensively cultured crustacean worldwide. The viral pathogens, Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) are responsible for causing severe mortalities in the hatchery and nursery phases. This study investigates the protection of postlarvae of freshwater against white tail disease (WTD) using plant extract derived from Cyanodon dactylon and the modulation of the prawn non‐specific immunity. To determine the immunomodulatory effect of C. dactylon extract, the prawn was injected with plant extract and various immunological parameters were estimated. The immunological parameters such as proPO, SOD, THC and clotting time were found to be significantly higher in the plant extract‐injected prawn when compared with control groups. The results of real time PCR analysis revealed up regulation on the expression proPO, SOD and lysozyme genes in MrNV and XSV challenged prawn postlarvae treated with C. dactylon extract. Infectivity experiment showed high relative per cent survival in MrNV and XSV‐challenged prawn postlarvae treated with C. dactylon extract. These results strongly indicate that the administration of C. dactylon plant extract enhances immunity of the prawn. Based on the results, this study recommends that the immersion of postlarvae in C. dactylon plant extract is a potential prophylactic agent against WTD.     

Effects of salinity acclimation on the growth performance,osmoregulation and energy metabolism of the oriental river prawn,Macrobrachium nipponense (De Haan)

In this study, we evaluated the growth, osmoregulation and energy metabolism of the oriental river prawn, Macrobrachium nipponense, reared during 6 weeks with different salinities (0, 8, 14 and 22 g/L). The results showed that the haemolymph osmolality of M. nipponense increased with an increase in ambient osmotic pressure; the isosmotic point was 490 mOs/kg H₂O. The prawns showed a higher survival rate, weight gain rate and hepatopancreas index in salinity 14 g/L. Digestive enzymes were all affected by salinity, and the highest activities were observed in the salinity 14 g/L. The mRNA expression of Na⁺‐K⁺‐ATPase in gills and p53 in hepatopancreas were the highest in salinity 22 g/L. The expressions of heat shock protein 90 and glutathione S‐transferase genes in hepatopancreas were significantly higher in the salinity 8 g/L. Lipid metabolism‐related genes in hepatopancreas were significantly expressed in the salinity 14 g/L. The glucose‐6‐phosphatase gene in hepatopancreas was highly expressed in the salinity 8 and 22 g/L, and the expression of the ecdysone receptor gene in hepatopancreas was significantly higher in the salinity 14 g/L. The results showed that salinity 14 g/L could promote the growth of M. nipponense. However, higher salinity conditions may cause physiological damage, which provides a theoretical basis for brackish water culture of M. nipponense.     

Effect of l‐ascorbyl‐2‐polyphosphate supplementation on growth performance,body composition,antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of ascorbic acid (AsA), in the form of l ‐ascorbyl‐2‐polyphosphate (LAPP) on growth performance, body composition, antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp, Litopenaeus vannamei. Five practical diets (46% crude protein and 7.6% lipid) supplemented with graded levels of AsA (14.64, 48.55, 84.98, 308.36 and 639.27 mg kg^?1 diet) were fed to five replicate groups of L. vannamei (mean initial wet weight 0.57 g). No significant differences were found on growth performance among all treatments. However, whole body lipid content significantly decreased with dietary AsA levels increasing. Activities of total antioxidant capacity, glutathione reductase and glutathione peroxidase were significantly affected by dietary AsA levels. Shrimp fed LAPP‐free diet had higher malondialdehyde content than those fed the diets supplemented with LAPP. Dietary AsA levels higher than 308.36 mg kg^?1 diet increased the survival of shrimps after 1, 2 and 3 h of acute salinity change. Broken‐line regression analysis on survival after 3 h of salinity stress and second‐degree polynomial regression analysis on glutathione reductase data indicated that the optimal dietary AsA requirement of L. vannamei was estimated to be 306.39, 319.75 mg kg^?1 diet respectively.