Identification of haematological markers in shrimp health assessment from the immune profile of Fenneropenaeus indicus on β-1,3-glucan administration and white spot syndrome virus challenge

Haematological alterations in Fenneropenaeus indicus fed marine yeast glucan-incorporated diet and challenged with white spot syndrome virus were analysed. Adult F. indicus 16.45 ± 2.12 g (mean ± SD) were reared in 25 ‰ sea water and allowed to acclimate for a period of 7 days. Diet was prepared incorporating glucan (0.2 %) extracted from marine yeast, Debaryomyces hansenii S169. The shrimps were reared on experimental diet for 28 days and then challenged with white spot syndrome virus through oral administration. Random sampling was done (n = 6 shrimps) on 1st, 15th and 28th day of the experiment and on post-challenge day 2 (PCD 2) and PCD 7 for haematological analysis. The immune parameters viz. plasma protein, total haemocyte count, phenoloxidase activity, superoxide anion production, alkaline phosphatase and acid phosphatase activities were estimated in the haemolymph. Superoxide dismutase, catalase, glutathione peroxidase, hydroperoxide, conjugated dienes and malondialdehyde concentrations in the haemolymph of shrimps were also analysed. Analysis of variance showed that there are significant differences (p < 0.05) in the immune and antioxidant parameters in different treatment groups of F. indicus. The shrimps fed glucan-incorporated diet showed higher survival rate with comparatively lower accumulation of lipid peroxidation products. Correlation coefficients showed that all haematological variables except hydroperoxides and conjugated dienes exhibited positive correlation with the survival rate. The immune variables and antioxidant parameters exhibited a greater degree of correlation with each other. When multiple regression of survival rate on all immune parameters was considered, the amount of variability explained was 93 % (R Square = 0.932). When significant regression coefficients among the immune parameters were taken into account, it was found that total haemocyte count (p < 0.001), phenoloxidase activity (p < 0.05), alkaline phosphatase activity (p < 0.05) and plasma protein concentration (p < 0.05) together are explaining 84 % (R ² = 0.842) of the variability, indicating that these four parameters are mainly supporting the survival. The amount of variability explained by the antioxidant parameters was 94 % (R ² = 0.938). When significant regression coefficients among the antioxidant parameters were taken into account, it was found that superoxide dismutase activity (p < 0.01) and catalase activity (p < 0.05) together are explaining 87.4 % (R ² = 0.874) of the variability, indicating that these two antioxidants act as the major immune effectors supporting the survival in shrimps.     

Acute exposure to copper induces variable intensity of oxidative stress in goldfish tissues

Copper is an essential element, but at high concentrations, it is toxic for living organisms. The present study investigated the responses of goldfish, Carassius auratus, to 96 h exposure to 30, 300, or 700 μg L^?1 of copper II chloride (Cu²⁺). The content of protein carbonyls was higher in kidney (by 158%) after exposure to 700 mg L^?1 copper, whereas in gills, liver, and brain, we observed lower content of protein carbonyls after exposure to copper compared with control values. Exposure to copper resulted in increased levels of lipid peroxides in gills (76%) and liver (95–110%) after exposure to 300 and 700 μg L^?1 Cu²⁺. Low molecular mass thiols were depleted by 23–40% in liver and by 29–67% in kidney in response to copper treatment and can be used as biomarkers toxicity of copper. The activities of primary antioxidant enzymes, superoxide dismutase and catalase, were increased in liver as a result of Cu²⁺ exposure, whereas in kidney catalase activity was decreased. The activities of glutathione-related enzymes, glutathione peroxidase, glutathione-S-transferase, and glutathione reductase were decreased as a result of copper exposure, but glutathione reductase activity increased by 25–40% in liver. Taken together, these data show that exposure of fish to Cu²⁺ ions results in the development of low/high intensity oxidative stress reflected in enhanced activities of antioxidant and associated enzymes in different goldfish tissues.     

Inclusion of Amazonian Mauritia flexuosa fruit pulp as functional feed in the diet for the juvenile Pacific white shrimp Litopenaeus vannamei

This study investigated the effects of the inclusion of Mauritia flexuosa fruit pulp in the diet on the growth performance, economic benefit, muscle composition, total contents of carotenoids and flavonoids, chromaticity parameters, total antioxidant capacity, and lipid peroxidation in the hepatopancreas and muscle of the juvenile shrimp Litopenaeus vannamei (weighing 1.26 ± 0.023 g). Five diets (37% protein, 10% lipid) consisting of 0 (control), 1.25%, 2.50%, 5.00% and 10.00% (w/w) M. flexuosa fruit pulp were formulated and provided to the shrimps (n = 50 per tank, treatments in triplicate) for 31 days. Results showed that there were no significant differences between the treatments in terms of the growth capacity, chromaticity parameters, muscle composition and total flavonoid content of shrimps (p > .05). However, the carotenoid content in the muscle and hepatopancreas of the shrimps was significantly increased due to the dietary carotenoids present in M. flexuosa fruit pulp (p < .05). This increase in the carotenoid content directly favoured the increase in the antioxidant capacity in the hepatopancreas and muscle of the shrimps (R² = 0.99 and R² = 0.97 respectively; p < .05). There was also a direct relationship between the carotenoid content and the decrease in lipid peroxidation in the hepatopancreas (R² = 0.90; p < .05). The feed cost kg^?1 in terms of weight gain and protein production decreased with the inclusion of high levels of M. flexuosa fruit pulp, reaching 8.11% and 18.76% of cost reduction, respectively, with the addition of 5.00% of M. flexuosa fruit pulp. Therefore, the inclusion of M. flexuosa fruit pulp in the diets of shrimp can increase the carotenoid content and the antioxidant capacity in the hepatopancreas and muscle, reduce lipid peroxidation in the hepatopancreas, and probably provide economic benefits for the rearing of juvenile shrimps.     

Effects of dietary inorganic copper on growth performance and immune responses of juvenile beluga,Huso huso

A 12‐week feeding trial was conducted to evaluate the optimum dietary inorganic copper (copper sulphate) in juvenile beluga, Huso huso. Eight semi‐purified diets containing 1.1 (Cu_1.0), 3.5 (Cu_4.0), 7.1 (Cu_7.0), 9.7 (Cu₁₀), 13.1 (Cu₁₃), 25.1 (Cu₂₅), 49.9 (Cu₅₀) and 195 (Cu₁₉₅) mg Cu kg^?1 diet in the form of CuSO₄.5H₂O were fed to fish of initial body weight 8.49 ± 0.32 g and length 11.85 ± 0.66 cm (mean ± SD) in triplicate groups in a flow‐through system. Weight gain (WG) of fish fed Cu₁₀ and Cu₁₃ diets was significantly higher than that of fish fed Cu_1.0, Cu_4.0, Cu₂₅, Cu₅₀ and Cu₁₉₅ diets (P < 0.05). Whole‐body and muscle crude protein increased with dietary Cu up to the supplementation level of 13.1 mg kg^?1 diet and then decreased. Whole‐body lipid content was negatively correlated, while whole‐body ash was positively correlated with dietary copper concentration. Hepatic copper–zinc superoxide dismutase activity of fish fed Cu₁₀ and Cu₁₃ diets was significantly higher than that of fish fed Cu_1.0, Cu_4.0 and Cu₁₉₅ diets. Hepatic thiobarbituric acid‐reactive substances of fish fed Cu₁₃ diet was significantly lower than those of fish fed the other diets except for that of fish fed Cu₁₀ diet. Aspartate aminotransferase, alanine aminotransferase and copper accumulation in tissues increased with dietary copper. Broken‐line analysis of WG suggested that the optimum dietary Cu level was 10.3 mg Cu kg^?1 diet. Therefore, these results may indicate that the optimum dietary Cu levels could be greater than 10.3 mg Cu kg^?1 diet but less than 13.1 mg Cu kg^?1 diet in juvenile beluga, when copper sulphate is used as the dietary source of inorganic copper.     

The role of açaí (Euterpe oleracea Mart. 1824) as a chemoprotective agent in the evaluation of antioxidant defence,oxidative damage and histology of juvenile shrimp Litopenaeus vannamei (BOONE, 1931) exposed to ammonia

This study investigated the effects of the use of the inclusion of açaí on the diet of shrimp Litopenaeus vannamei on antioxidant and histopathological responses after exposure to ammonia. The shrimps were fed two experimental diets: control and with 10% of açaí inclusion (W/W), for 35 days. Afterwards, the organisms were exposed at four concentrations of ammonia (0.01‐control; 0.26; 0.48 and 0.91 mg NH₃‐N L^?1) for 96 hr. The total antioxidant capacity (ACAP) of the gills decreased significantly in both diets when exposed to ammonia, whereas in the muscle, the açaí promoted an increase in ACAP. Concomitantly, lipid peroxidation levels increased in the gills and decreased in muscle. After exposure to ammonia, glutathione‐S‐transferase activity increased in hepatopancreas in shrimps fed with açaí facilitating the detoxification of lipid peroxidation by‐products, and the concentration of protein sulfhydryl groups decreased in the gills and muscle of the shrimp of the control diet, evidencing protein damage, an unobserved response in shrimps that received the açaí diet. Histopathological changes decreased in açaí‐fed shrimps about the control diet after exposure to ammonia. It is concluded that açaí mitigated ammonia‐induced histopathological changes, improved the antioxidant defence system (gills and muscle) and attenuated the lipid damage in the muscle.     

Effect of calcium and magnesium hardness on acute copper toxicity to Indian major carp, Labeo rohita (Hamilton) and catfish, Channa punctatus (Bloch)

Four experiments were conducted to evaluate the effects of calcium and magnesium hardness on the acute toxicity of copper sulphate to Indian major carp, rohu (Labeo rohita, Hamilton) fingerlings and juvenile catfish (Channa punctatus, Bloch) in medium alkalinity experiments. A preliminary bioassay determined the 96 h LC₅₀ of copper sulphate to be 0.56 mg L^?1 for L. rohita fingerlings and 11.78 mg L^?1 for juvenile C. punctatus placed in water with calcium hardness and total alkalinity set at 100 mg L^?1 CaCO₃. In the first experiment, rohu were exposed to 0.56 mg L^?1 copper sulphate in environments where calcium hardness was varied from 50 to 350 mg L^?1 CaCO₃ and total alkalinity was 100 mg L^?1 CaCO₃. As calcium hardness increased, copper‐induced rohu mortalities decreased significantly from 90% at 50 mg L^?1 CaCO₃ to 7% at 350 mg L^?1 CaCO₃. In the second experiment, rohu were exposed to 0.56 mg L^?1 copper sulphate in environments where magnesium hardness was varied from 50 to 350 mg L^?1 CaCO₃ with total alkalinity set at 100 mg L^?1 CaCO₃. Hundred percent mortality was observed in magnesium‐based hardness treatments. In the third experiment, catfish were exposed to 11.78 mg L^?1 copper sulphate in environments where calcium hardness was varied from 50 to 400 mg L^?1 and total alkalinity was 100 mg L^?1 CaCO₃. As calcium hardness increased, copper‐induced catfish mortalities decreased significantly from 90% at 50 mg L^?1 CaCO₃ to 4% at 400 mg L^?1 CaCO₃. In the fourth experiment, catfish were exposed to 11.78 mg L^?1 copper sulphate in environments where magnesium hardness was varied from 50 to 400 mg L^?1 CaCO₃, with total alkalinity set at 100 mg L^?1 CaCO₃. In this case, 100% mortality was also observed in magnesium‐based treatments. Mortality rates in magnesium hardness treatments were consistent with those in the second experiment. These data suggest a calcium‐specific mechanism with respect to acute copper toxicity both in rohu and catfish.     

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.     

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.     

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.     

Oxidative stress and antioxidant responses in juvenile Brazilian flounder <Emphasis Type="Italic">Paralichthys orbignyanus</Emphasis> exposed to sublethal levels of nitrite

This study evaluated the effects of short-term exposure to sublethal levels of nitrite on oxidative stress parameters and histology of juvenile Brazilian flounder Paralichthys orbignyanus. An assessment of fish recovery was also performed. Fish were exposed to 0.08 (control), 5.72, 10.43, and 15.27 NO₂-N mg L^?1 for 10 days followed by the same recovery time. Gill, liver, and muscle samples were collected after 1, 5, and 10 days of exposure and after recovery for the measurement of antioxidant capacity against peroxyl radicals (ACAP), glutathione-S-transferase (GST) activity, content of non-protein (NPSH) and protein thiols (PSH), and lipid peroxidation levels by thiobarbituric acid-reactive substances (TBARS) content. Nitrite exposure induced alterations which compromised the overall antioxidant system (reduced ACAP and GST activity) and enhanced oxidative damage in lipids and proteins. Increases in GST activity and NPSH and PSH contents were also demonstrated. The recovery period allowed for resumption of basal levels for all (treatment 5.72 NO₂-N mg L^?1) or some of the evaluated parameters (other treatments). In conclusion, exposure to nitrite concentrations from 5.72 to 15.27 NO₂-N mg L^?1 induced oxidative stress and antioxidant responses in juvenile Brazilian flounder. The 10-day recovery period was sufficient for a complete resumption of basal physiological condition of fish exposed to concentrations of up to 5.72 NO₂-N mg L^?1.     

Effects of different dietary copper sources on the growth and intestinal microbial communities of Pacific white shrimp (Litopenaeus vannamei)

An 8‐week feeding trial was conducted to evaluate the effects of dietary copper sources on growth performance and intestinal microbial communities of juvenile Pacific white shrimp (Litopenaeus vannamei). Four isonitrogenous (420 g/kg crude protein) and isolipidic (70 g/kg crude lipid) experimental diets were formulated to contain different copper sources, Diet NSC (no‐supplemented copper), Diet CS (copper sulphate), Diet Availa (copper amino acid complex from Availa^®Cu100) and Diet M (1:1, copper sulphate + copper amino acid complex). A total of 360 Pacific white shrimp juveniles (initial weight: 1.86 ± 0.03 g) were randomly allocated into four groups, with three replicates per group and 30 shrimps each replicate. An Illumina‐based sequencing method was used to analyse the intestinal bacterial composition of shrimp juveniles. The results revealed that shrimps fed Diet M had significantly higher percent weight gain (PWG) than that fed Diet NSC and Diet CS, while there were no significant differences between Diet M and Diet Availa. Survival was not affected by the dietary copper sources. Proteobacteria and Bacteroidetes were dominant in the intestines of Pacific white shrimp. Shrimps fed Diet M and Diet Availa had higher bacterial richness and diversity than those fed Diet NSC and Diet CS with no significant differences among all treatments. Furthermore, Diet M and Diet Availa which contained copper amino acid complex reduced the relative abundance of Vibrionaceae in shrimp intestinal tract. This study indicated that dietary copper sources could affect the intestinal microbial communities and diets supplemented with Availa^®Cu100 could have a positive impact on the growth performance and decrease the potential risk of disease in Pacific white shrimp.     

Antioxidant effects of nanoencapsulated lipoic acid in tissues and on the immune condition in haemolymph of Pacific white shrimp Litopenaeus vannamei (Boone, 1931)

The antioxidant and oxidative damage responses and immune parameters in shrimp, Litopenaeus vannamei, reared in clear water (CW) and in a biofloc system (BFT) were evaluated. The animals were given feed supplemented with either nanocapsules containing the antioxidant lipoic acid (NCLA) or empty nanocapsules (NC). Antioxidant and oxidative stress analyses were evaluated by measuring glutathione S‐transferase activity (GST), reduced glutathione (GSH) concentration and lipid peroxidation (TBARS) in the gills, hepatopancreas and muscle. The immune parameters were then determined by the differential hemocyte count in the haemolymph. The final weight of the shrimp reared in both CW and BFT and treated with NCLA increased (p < .05). The shrimps reared in BFT showed increased GST activity in the gills and hepatopancreas after NCLA treatment (p < .05). Higher GSH levels were observed in the hepatopancreas with NCLA supplementation in CW and BFT (p < .05). Decreased levels of TBARS were observed in the gills and muscles of shrimp supplemented with NCLA and maintained in CW (p < .05). A decrease in the percentage of hyaline hemocytes and an increase in granular hemocytes were observed in shrimp treated with NCLA in CW and BFT (p < .05). The antioxidant treatment induced positive responses in terms of the zootechnical, biochemical and immunological parameters.     

Antioxidant vitamins, minerals and lipid levels in diets for Atlantic salmon (Salmo salar, L.): effects on growth performance and fillet quality

An experiment with 2^{(7 ? 3)} reduced factorial design was conducted to study the biological effects of pro‐ and antioxidant micronutrients and lipid in Atlantic salmon. Vitamins C and E, astaxanthin, lipid, iron, copper and manganese were supplemented at high and low levels. For vitamins and minerals, high levels were chosen to be below the anticipated toxic level and the low levels were just above the requirement (vitamin C, 30 and 1000 mg kg^?1; vitamin E, 70 and 430 mg kg^?1; Fe, 70 and 1200 mg kg^?1; Cu, 8 and 110 mg kg^?1; Mn, 12 and 200 mg kg^?1). For astaxanthin, the dietary levels were 10 and 50 mg kg^?1 and for lipid, 150 and 330 g kg^?1. The experiment was started with postsmolts (148 ± 17 g) and lasted for 5 months. The variation in micronutrients had only minor effects on growth, feed conversion and fillet quality, measured as lipid and astaxanthin deposition. High dietary lipid had a profound positive effect on growth and feed conversion but gave fillets nearly two times the fat content that was found in fish fed the low lipid diet. Astaxanthin deposition in the fillet was primarily affected by dietary astaxanthin with a positive effect of high dietary lipid in week 14 but not in week 23. Vitamin E protected the fillet against iron ascorbate stimulated oxidation, with no effect of the other nutrient variables.     

Effects of Bacillus preparations on immunity and antioxidant activities in grass carp (Ctenopharyngodon idellus)

This experiment was conducted to study the effects of Bacillus preparations on immunity and antioxidant activities in grass carp. A total of 315 grass carp, with similar initial weight (average weight of fish 45?g), were randomly divided into three groups with three replicates. The control group was fed the basal diet without Bacillus; treatment group 1 was added Bacillus preparation no. 1 with 1?×?10⁸?cfu/m³ per 7?days in culture water and also fed the basal diet; treatment group 2 was fed the basal diet mixed with 0.5?% Bacillus preparation no. 2, and the culture water was added 1?×?10⁸?cfu/m³ Bacillus preparation no. 1 per 7?days. After 4?weeks of culture, 12 grass carp from each replicate were taken randomly for the determination of immune response and oxidization resistance indices. The results showed that compared with control, the level of globulin and IgM of treatment group 2 was significantly increased (P?<?0.05), which was also significantly higher (P?<?0.05) than that of group 1. For the non-specific immunity index, compared with control, the lysozyme activity and complement (C3) content of treatment group 1 significantly increased (P?<?0.05), and the level of myeloperoxidase and C3 of treatment group 2 was significantly higher (P?<?0.05) than that of control, which was also significantly higher (P?<?0.05) than that of group 1. In the serum, compared with the control, the level of total antioxidant activity (T-AOC), antisuperoxide anion free radical (ASAFR) and glutathione (GSH) of two treatment groups was significantly increased (P?<?0.05); GSH content of treatment group 2 was significantly (P?<?0.05) higher than that of treatment group 1 and control. There was no significant difference between T-AOC and ASAFR among the two treatment groups; no significant effect was found on glutathione peroxidase (GSH-Px) and maleic dialdehyde (MDA) among the three groups. In the liver, T-AOC, SOD, ASAFR, GSH-Px and GSH from two treatment groups were significantly higher (P?<?0.05) than those of control; T-AOC, SOD and catalase from treatment group 2 were significantly higher (P?<?0.05) than those of treatment group 1; and MDA of two treatment groups was significantly decreased (P?<?0.05) as compared with the control. The results indicate that Bacillus preparations added into water can increase serum immunoglobulin levels and most of non-specific immune parameters content and enhance the antioxidant ability of grass carp, while adding Bacillus preparation into the water and feed is much better.     

Effects of nucleotides on growth performance,immune response,disease resistance and intestinal morphology in shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> fed with a low fish meal diet

Nucleotides (NT) are widely used as an immunostimulant in aquaculture. A 10-week feeding trial was conducted to evaluate the effects of dietary NT on the growth performance, immune response, disease resistance and intestinal morphology of the Pacific white shrimp Litopenaeus vannamei (initial mean body weight: 0.39 ± 0.00 g). Five isolipidic (about 7 % crude lipid) and isonitrogenous (about 39 % crude protein) practical diets were supplemented with graded levels of NT (0, 60, 90, 120 and 1200 mg kg^?1), respectively. These diets were named as N1, N2, N3, N4 and N5. Each diet was randomly fed to six tanks of shrimps, and each tank contained 40 shrimps. The shrimps were fed four times daily (07:00, 11:00, 16:00 and 21:00 h). The results showed that there were no significant differences in survival, final weight, specific growth rate, feed intake, feed conversion ratio and whole-body compositions among all the treatments (P > 0.05). Shrimps fed the control diet had the significantly lowest activities of superoxide dismutase, total nitric oxide synthase and lysozyme (P < 0.05). However, those parameters increased when dietary NT increased from 60 to 120 mg kg^?1 and decreased as inclusion level increased to 1200 mg kg^?1 (P < 0.05). The cumulative mortality of the shrimps challenged with Vibrio parahaemolyticus was significantly higher in the treatments without dietary NT supplementation than those in treatments with dietary NT (≥90 mg kg^?1) (P < 0.05). The jejunum wall thickness in shrimps fed the control diet was significantly lower than that in the treatments of N2, N3 and N4 (P < 0.05). For the villus height, the highest value (49.29 µm) was found in N3. In summary, the present study showed that 90 mg kg^?1 of dietary NT is the optimum dietary level for good gut health, immune response and disease resistance of Pacific white shrimp fed a diet with 18 % fish meal.     

Effects of dietary nucleotides on the antioxidant status and serum lipids of rainbow trout (Oncorhynchus mykiss)

An 8‐week feeding experiment was conducted to evaluate the effects of dietary nucleotides (0, 0.5, 1, 1.5 and 2 g kg^?1) on serum antioxidative biomarkers and lipid contents of rainbow trout fingerlings. The serum triglycerides and low‐density lipoprotein (LDL)‐C levels were significantly decreased in fish fed diets containing 1.5 and 2 g kg^?1, whereas serum high‐density lipoprotein (HDL)‐C was higher in trout receiving diets with those levels of nucleotides. A significant decrease in thiobarbituric acid reactive substances concentration was observed in groups receiving 1.5 or 2 g kg^?1 of dietary nucleotides in comparison with control group (?32% and ?27%, respectively). A significant decrease was observed in catalase and serum glutathione peroxidase activity in trout fed 1.5 and 2 g kg^?1 dietary nucleotides compare with other groups. There were no significant changes in serum superoxide dismutase activity among the experimental groups. In conclusion, these results suggest that dietary nucleotides may reduce lipid peroxidation by increasing the levels of antioxidant lipoprotein, HDL‐C, and decreasing the LDL‐C, the main substrate of lipid peroxidation.     

Redox dysregulation and bioaccumulation of copper in tilapia Oreocromis niloticus

The objective of this study was to evaluate the effect of two concentrations of copper (0.13 and 0.5 mg L^?1) on juveniles of tilapia, Oreocromis niloticus (Linnaeus), using two toxicological assays (I‐ copper exposure and II‐ copper exposure followed by recovery). Doses were taken from the liver of the test organisms to determine the concentrations of copper, reduced glutathione, activities of glutathione S‐transferase and catalase, and lipid peroxidation. Exposure to copper in both assays resulted in significant accumulation of this metal in the analysed tissue and in decreased concentration of reduced glutathione. There was no alteration in the activities of catalase and glutathione S‐transferase and in lipid peroxidation.     

Assessment of Aquaflor(®) , copper sulphate and potassium permanganate for control of Aeromonas hydrophila and Flavobacterium columnare infection in sunshine bass, Morone chrysops female × Morone saxatilis male

Two experiments were conducted to test the effectiveness of different therapeutants against a mixed infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass. Experiment 1 evaluated copper sulphate, florfenicol‐medicated feed and potassium permanganate (KMnO₄) against a natural mixed infection. Experiment 2 further evaluated copper sulphate as a treatment to control an experimental mixed infection. In experiment 1, naturally infected untreated fish had the lowest final survival per cent, at 71%, while florfenicol‐medicated feed at 15 mg kg^?1 body weight for 10 days or copper sulphate at 2.1 mg L^?1 (1% of the total alkalinity) for 24 h produced the highest final survivals, at 90% and 88%, respectively. The final survival of the naturally infected fish administered florfenicol‐medicated feed was significantly different (P < 0.1) from the untreated fish. The survival curves for the florfenicol and the copper sulphate at 2.1 mg L^?1 were significantly improved from the untreated fish. In experiment 2, fish were challenged by waterborne exposure to A. hydrophila and F. columnare and either not treated or treated with copper sulphate at 2.1 mg L^?1. At the end of experiment 2, the per cent survival of the challenged fish treated with copper sulphate (99%) was significantly higher (P < 0.05) than the non‐treated (61%). The results illustrate clear benefit of florfenicol and copper sulphate against a mixed infection of A. hydrophila and F. columnare.     

Predation and energetics of Penaeus indicus (Decapoda: Penaeidae) larvae feeding on Brachionus plicatilis and Artemia nauplii

Brachionus plicatilis and Artemia nauplii were fed to a number of larval stages of the penaeid prawn Penaeus indicus to determine ingestion rates, larval energy requirements and to establish at which stage larval predation commenced. The raptorial feeding rates were then contrasted on an energy basis with filter feeding rates for P. indicus larvae to compare the relative efficiency of these two feeding mechanisms. Brachionus was first eaten as early as protozoea 1 to protozoea 2, while the maximum ingestion rate of 300 rotifers larva^?1 d^?1 (1.06 J larva^?1 d^?1) was obtained during protozoea 3 to mysis 1. Artemia were effectively ingested by P. indicus protozoea 3 (4.1 J larva^?1 d^?1) to post-larva (8.2 J larva^?1 d^?1). Daily energy intake rate from filter feeding increased from 1.1 J larva^?1 d^?1 during protozoea 1 to reach a peak of 5.32 J larva^?1 d^?1 during mysis 3 after which it declined to 2.66 J larva^?1 d^?1 during the post-larval stage. This decline in energy intake from filter feeding with a concomitant increase in energy intake from Artemia predation demonstrates a predominant feeding mode changeover point during mysis 3. Energy intake was consistently low with Brachionus, indicating that it may be unnecessary for commercial culture purposes.     

Effects of dietary aflatoxin B1 on growth performance,body composition,haematological parameters and histopathology of juvenile Pacific white shrimp (Litopenaeus vannamei)

An 8‐week feeding trial was conducted to evaluate the effects of dietary aflatoxin B₁ (AFB₁) on growth performance, haematological parameters and histological changes in juvenile Pacific white shrimp, Litopenaeus vannamei. Six practical diets (455 g kg^‐1 protein, 78 g kg^‐1 lipid) with different levels of AFB₁ (0, 25, 50, 100, 500, 1000 μg kg^?1) were formulated. Each diet was fed to triplicate groups of shrimps (initial weight: 0.52 g). The results showed that shrimp fed with control diet (0 μg kg^?1 AFB₁) had significant higher weight gain (WG) and specific growth rate (SGR) than other groups. However, there were no significant differences in feed efficiency (FE) or hepatosomatic index (HSI) among all groups. Compared to the control diet, AFB₁ supplementation significantly changed the activities of shrimp serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total antioxidant capacity (T‐AOC) and glutathione S‐transferase (GST) and the content of cholesterol (CHO). Histological damages were identified in the hepatopancreas of shrimp when dietary AFB₁ level was over 107.6 μg kg^?1. Based on this study, it was concluded that the AFB₁ level in Pacific white shrimp diet should be <38.1 μg kg^?1.