Effects of graded levels of dietary myo‐inositol on non‐specific immune and specific immune parameters in juvenile Jian carp (Cyprinus carpio var. Jian)

The aim of this study was to evaluate the effects of myo‐inositol (MI) on non‐specific immune and specific immune defence in fish. A total of 1050 Jian carp (Cyprinus carpio var. Jian) (22.28±0.07 g) were randomly distributed into seven groups, of three replicates each, of feeding diets containing graded levels of MI (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg MI kg^?1 diet). After a 60‐day growth trial, an infectious trial was conducted by injection of Aeromonas hydrophila for 17 days. Results showed that the red blood cell (RBC) and the white blood cell count were significantly increased with increasing MI levels up to 535.8 mg kg^?1 diet (P<0.05). The spleen index showed a tendency similar to RBC, whereas the head kidney index showed the inverse pattern (P<0.05). The phagocytic activity of leucocytes, haemagglutination titre, lysozyme activity, anti‐A. hydrophila antibody titre and immunoglobulin M, after being injected with A. hydrophila, were all improved with an increase in the MI levels up to 232.7–687.3 mg kg^?1 diet respectively (P<0.05). Myo‐inositol did not influence serum acid phosphatase activity and total iron‐binding capacity (P>0.05). These results suggested that MI could enhance non‐specific immune and specific immune responses in fish.     

Effects of dietary pyridoxine on disease resistance,immune responses and intestinal microflora in juvenile Jian carp (Cyprinus carpio var. Jian)

This experiment was conducted to evaluate the effects of dietary pyridoxine on disease resistance, immune responses and intestinal microflora of fish. A total of 1050 Jian carp (11.71 ± 0.05 g) were randomly distributed into seven groups, feeding diets containing graded levels of pyridoxine (0.2, 1.7, 3.2, 5.0, 6.3, 8.6 and 12.4 mg kg^?1 diet). After 80 days of feeding, a challenge trial was conducted by injection of Aeromonas hydrophila for 17 days. Results indicated that with increasing dietary pyridoxine concentration up to 5.0 mg kg^?1 diet, survival rate after challenge with A.hydrophila and phagocytic activity of leukocyte were improved (P < 0.05), and plateaued thereafter (P > 0.05). Red blood cell and white blood cell counts were lowest when fed the diet containing 1.7 mg pyridoxine kg^?1 diet. Haemagglutination titre, lysozyme activity, acid phosphatase activity, total iron‐binding capacity, antibody titre and immunoglobulin M content followed the similar pattern to that observed with survival rate. Aeromonas hydrophila, Escherichia coli and Lactobacillus counts in intestine were not affected by dietary pyridoxine concentration (P > 0.05). These results suggested that pyridoxine could enhance immune response of fish.     

Immune response,disease resistance and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of pantothenic acid

This study was to investigate the effect of dietary pantothenic acid (PA) on the disease resistance, immune response and intestinal microflora on juvenile Jian carp (Cyprinus carpio var. Jian). Seven diets (4.0, 15.5, 25.6, 36.1, 45.9, 56.1 and 65.9 mg PA kg^?1) were fed to Jian carp (12.95 ± 0.03 g) for 9 weeks. After 9‐week feeding trial, the challenge experiment with Aeromonas hydrophila was conducted to determine the impact of PA on fish disease resistance. Survival rate after challenge was promoted with the increasing PA levels (P < 0.05). Blood counts also significantly increased up to the dietary PA level of 25.6 mg PA kg^?1 (P < 0.05). Leucocyte phagocytic activity, lectin potency, lysozyme and acid phosphatase activity, and total iron‐binding capacity were improved with increasing PA levels (P < 0.05). Serum immunoglobulin M level and agglutination antibody titre to A. hydrophila were increased (P < 0.05) in fish fed the diets with the dietary PA levels between 56.1 and 65.9 mg kg^?1. PA also promoted the growth and reproduction of Lactobacillus and depressed Escherichia coli and A. hydrophila (P < 0.05). These results suggested that pantothenic acid could improve disease resistance, immune response, and the balance of intestinal microflora in juvenile Jian carp.     

Extracts of Oliviera decumbens and Satureja khuzestanica as immunostimulants affect some innate immunity indices of Cyprinus carpio against Aeromonas hydrophila infection

In this study, the effect of extracts of two herbs (Oliviera decumbens and Satureja khuzestanica) on immune response of carp was investigated. At the beginning of experiment, fish were divided to two groups including vaccinated (using vaccine developed against A. hydrophila) and non‐vaccinated. Fish in both groups were fed diets containing O. decumbens and S. khuzestanica and combination of two herbal extracts for 5 weeks. Control fish (negative control) and fish vaccinated only (positive control) were fed basal diets without supplements of herbal extracts. Lysozyme activity, antibody titre, complement activity and bactericidal activity in serum were measured. After 5 weeks feeding, fish were infected with A. hydrophila and mortalities were recorded. In both experimental groups, no significant differences were found in terms of alternative haemolytic complement (ACH50) activity and antibody titres of Serum. In non‐vaccinated fish group, lysozyme and bactericidal activity of fish fed S. khuzestanica or combination of O. decumbens and S. khuzestanica was higher compared with control and other experimental treatments. In vaccinated fish group, the lysozyme and bactericidal activity was not significant in all treatments compared with control group. The results of this study showed that feeding non‐vaccinated and vaccinated carp with only S. khuzestanica or in combination with O. decumbens enhance only some immunity indices including lysozyme and bactericidal activity in non‐vaccinated fish and these extracts have no immunological stimulatory role on vaccinated individuals.     

Requirements of vitamin C (L-ascorbyl-2-monophosphate-Mg and L-ascorbyl-2-monophosphate-Na) and its effects on immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to evaluate two vitamin C derivatives, L‐ascorbyl‐2‐monophosphate‐Mg (C2MP‐Mg) and L‐ascorbyl‐2‐monophosphate‐Na (C2MP‐Na), to satisfy the vitamin C requirement and to test their effects on the immune responses of juvenile grouper, Epinephelus malabaricus. C2MP‐Mg and C2MP‐Na were each supplemented at 20, 50, 80, 150, 250, and 400 mg kg^?1 diet in the basal diet providing of 7, 18, 31, 51, 93, 145 mg ascorbic acid (AA) equivalent of C2MP‐Mg kg^?1 diet and 4, 10, 17, 31, 47, 77 mg ascorbic acid (AA) equivalent of C2MP‐Na kg^?1 diet, respectively. Basal diet without AA supplementation was included as control. Each diet was fed to triplicate groups of grouper (mean initial weight 3.20 ± 0.05 g). Fish fed diets supplemented with either C2MP‐Mg or C2MP‐Na had significantly (P < 0.05) greater weight gain (WG), feed efficiency and survival than those fed the unsupplemented control diet. Liver ascorbate concentrations in fish generally increased as dietary C2MP‐Mg or C2MP‐Na supplementation level increased. Haemolytic complement activity was higher in fish fed diets supplemented with 92 mg AA equivalent of C2MP‐Mg kg^?1 or 10–17 mg AA equivalent of C2MP‐Na kg^?1 than fish fed the unsupplemented control diet. Lysozyme activity was higher in fish fed ≥51 mg AA equivalent of C2MP‐Mg kg^?1 or ≥47 mg AA equivalent of C2MP‐Na kg^?1 than fish fed the unsupplemented control diet. Analysis by broken‐line regression of WG indicated that the adequate dietary vitamin C concentration from each vitamin C derivative in growing grouper is 17.9 mg AA equivalent of 2MP‐Mg kg^?1 and 8.3 mg AA equivalent of C2MP‐Na kg^?1, and it also indicated that C2MP‐Mg is about 46% as effective as C2MP‐Na in meeting the vitamin C requirement of grouper.     

Vaccination strategies with oral booster for surubim hybrid (Pseudoplatystoma corruscans × P. reticulatum) against haemorrhagic septicaemia

This study evaluated the efficiency of differently prepared vaccines against Aeromonas hydrophila in the hybrid surubim (Pseudoplatystoma corruscans × P. reticulatum). Survival and haemato‐immunological parameters were compared between the treatments: non‐vaccinated fish (C); bacterin‐vaccinated fish (B); bacterin plus oral booster vaccinated fish (B+O); bacterin and toxoid‐vaccinated fish (B+T) and bacterin, toxoid and oral booster‐vaccinated fish (B+T+O). Fourteen‐days vaccinated fish from B+O and B+T+O were fed with an oral booster for 4 days. After 1 week, the fish were intraperitoneally challenged with 2 × 10⁸ CFU mL^?1 of A. hydrophila. Fish from the treatment B+T+O showed the lowest cumulative mortality (11.36%) 96 h after challenge, compared with other treatments (22.72–44.04%), and a relative survival of 74%. Serum immunoglobulin in B+T+O fish was higher than in other treatments. All vaccinated fish showed an increased agglutination titre when compared with non‐vaccinated fish, both before and after challenge. Fish fed with oral booster showed an increase in phagocytic percentage before and after challenge. It can be inferred that the oral booster vaccination was efficient in reducing mortality in hybrid surubim by enhancing the response against haemorrhagic septicaemia due to A. hydrophila infection.     

Effects of different dietary protein and lipid levels on growth, feed utilization and body composition of red porgy (Pagrus pagrus) fingerlings

Two feeding trials were conducted to determine the minimum dietary protein level producing maximum growth, and the optimum protein to energy ratio in diets for red porgy (Pagrus pagrus) fingerlings, respectively. In the first trial, six isoenergetic diets were formulated with protein levels ranging from 400 to 650 g kg^?1 in increments of 50 g kg^?1, and fed for 11 weeks to 2.8 g average initial weight fish. Weight gain, specific growth rate and feed efficiency were significantly higher with diets containing higher protein levels, when compared with dietary levels below 500 g kg^?1. The highest protein efficiency ratios were obtained in fish fed 500 g kg^?1 dietary protein. The minimum dietary protein level producing maximum fish growth was found to be 500 g kg^?1. In the second trial, 15 g average initial weight fish were fed for 12 weeks, six diets containing three different lipid levels (100, 150 and 200 g kg^?1) combined with two protein levels (450 and 500 g kg^?1). Weight gain values increased when dietary lipids increased from 100 to 150 g kg^?1, with a further decrease for 200 g kg^?1 lipids in diets; the lowest fish growth being supported by 200 g kg^?1 dietary lipids. Fish growth was significantly higher when dietary protein increased from 450 to 500 g kg^?1. There was no evidence of a protein‐sparing effect of dietary lipids. Liver protein and lipid contents were low when compared with other fish species. All diet assayed produced high liver glycogen accumulation. The recommended protein and lipid levels in diets for red porgy fingerlings were 500 and 150 g kg^?1, respectively.     

The pigmenting effect of different carotenoids on fancy carp (Cyprinus carpio)

The optimal concentration of a panel of individual and combined carotenoid sources on skin pigmentation in fancy carp was investigated by nine experimental diets that were formulated and supplemented with astaxanthin at 25 mg kg^?1, lutein at 25 and 50 mg kg^?1, β‐carotene at 25, 50 and 75 mg kg^?1, and lutein combined with β‐carotene at 25 : 25 and 50 : 50 mg kg^?1, while a diet without supplemented carotenoid served as a control. The results showed that serum TC of fish fed diets containing supplemented with lutein plus β‐carotene at 25 : 25; 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet were higher than the other treatments (P ≤ 0.05). Serum TC of the respective treatments was 6.2 ± 2.0, 7.8 ± 3.3 and 7.3 ± 1.9 μg mL^?1 serum, respectively. Fish fed diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet had serum astaxanthin concentrations similar to fish fed the diet with astaxanthin alone at 25 mg kg^?1. Serum astaxanthin concentrations was 0.7 ± 0.01, 0.9 ± 0.01, 0.4 ± 0.02 and 1.7 ± 0.18 μg mL^?1 serum, respectively. The chromaticity of fish body skin of red and white position was assessed by colourimetry using the CIE L*a*b (CIELAB) system. Pigmentation response of skin redness of fancy carp fed with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were higher than other treatments (P ≤ 0.05) but they were similar to fish fed with 25 mg kg^?1 astaxanthin diet. The redness (a* values) of fish fed diets with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were 28.3 ± 0.53, 29.9 ± 1.38, 28.8 ± 3.95 and 28.5 ± 2.49, respectively. After 3 weeks of feeding the experimental diets, the fish fed on a diet without carotenoid supplement for one week demonstrated that the same three groups still retained their redness and had an overall tendency to improve skin colouring. Finally, concentrations 50 mg kg^?1 of lutein, or the combination of lutein and β‐carotene at 25 : 25 mg kg^?1 showed the highest efficiency for improving skin pigmentation and redness of skin.     

Effect of dietary thiamin supplement on immune responses and intestinal microflora in juvenile Jian carp (Cyprinus carpio var. Jian)

This study was to investigate the effect of dietary thiamin on the immune response and intestinal microflora in juvenile Jian carp (Cyprinus carpio var. Jian). Seven diets (0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg thiamin kg^?1) were fed to Jian carp (8.20 ± 0.02 g). At the end of feeding trial, red blood cell counts and white blood cell counts increased with increasing dietary thiamin levels up to 0.79 and 1.06 mg kg^?1 diet, respectively (P < 0.05); Lactobacillus counts in intestine increased gradually, and maximum values obtained when the thiamin level was at 0.79 mg kg^?1 diet (P < 0.05). After the feeding trial, fish were injected with Aeromonas hydrophila only one time on the first day of the challenge trial and fed the same diets as the growth trial for 17 days. The survival rate, leucocyte phagocytic activity, lectin potency, acid phosphatase activity, lysozyme activity, total iron‐binding capacity and immunoglobulin M content of fish after being injected with A. hydrophila were all improved with increasing dietary thiamin levels up to a point, respectively (P < 0.05). These results suggested that thiamin could improve immune response and the balance of intestinal microflora of juvenile Jian carp.     

Protective effects of basil (Ocimum basilicum) ethanolic extract supplementation diets against experimental Aeromonas hydrophila infection in common carp (Cyprinus carpio)

This study evaluated the effects of basil (Ocimum basilicum) leaves extract supplement on growth, blood parameters and resistance to Aeromonas hydrophila in Cyprinus carpio fingerlings. Basil leaves were mixed thoroughly with feed at 0, 100, 200, 400, 800 and 1600 mg kg^?1 of diet and were fed in triplicate group for 2 months. At the end of this period, growth performance and survival were determined. Then fish were challenged intraperitoneally with A. hydrophila and mortalities were recorded up to 10 days post challenge. Results indicated that WBC, RBC, Ht, Hb, total protein, albumin and globulin were higher (P < 0.05) in fish fed diets containing O. basilicum compared with the control. Highest specific growth rate values were observed in 400 mg kg^?1 group. Feed conversion ratio was significantly lower (P < 0.05) in fingerlings fed 400 and 800 mg O. basilicum kg^?1 dry diet. After 10 days post challenge, total protein, WBC, RBC and Hb were significantly higher in 400 mg kg^?1 compared with other groups. These results reveal that a dietary O. basilicum leaves extract of 400 mg kg^?1 fed for 60 days leads to increased growth performance and survival rate as well as improved feeding efficiency in common carp fingerlings rendering them more resistant against infection by A. hydrophila.     

Live Spirulina (Arthrospira platensis) as a growth and immunity promoter for Nile tilapia, Oreochromis niloticus (L.), challenged with pathogenic Aeromonas hydrophila

The use of antibiotics to control fish disease is one of the constraints in tilapia farming. Hence, the use of probiotic and live organisms as alternative strategies has received much more attention. This study was undertaken to evaluate the use of Spirulina (Arthrospir platensis) as a growth and immunity promoter for Nile tilapia, Oreochromis niloticus (L.). Fish (1.88 g) were randomly distributed at 20 fish per 100 L aquarium and fed a diet containing either 0.0, 1.25, 2.5, 5.0, 7.5 or 10.0 g Spirulina kg^?1 diet for 12 weeks. After the feeding trial, fish of each treatment were challenged by pathogenic Aeromonas hydrophila, which was given by an interperitoneal injection, and they were kept under observation for 10 days to record any abnormal clinical signs and the daily mortality rate. The growth‐promoting influence of Spirulina was observed with fish, and the optimum growth and feed utilization were obtained at 5.0 g Spirulina kg^?1 diet. No significant changes were observed in fish survival among the different treatments. Spirulina supplementation improved the protein and lipid contents in fish bodies without significant differences among them, especially when fed 2.5–10.0 g kg^?1 diet, but no significant differences in the moisture and ash contents were observed among different treatments. The physiological parameters were improved when fish were fed a Spirulina‐enriched diet. However, the highest red blood cells, white blood cells and nitro blue tetrazolium values were obtained at 5.0–10.0 g Spirulina kg^?1 diet; meanwhile, the lowest value was obtained in the control. Total fish mortality due to A. hydrophila infection decreased with an increase in the Spirulina level in fish diets. These results indicate that Spirulina supplement is promising as an alternative method to antibiotics for disease prevention in tilapia aquaculture, and the optimum level of Spirulina in the fish diet should be 5.0–10.0 g kg^?1 diet.     

Effect of high dietary intakes of vitamin E and n-3 HUFA on immune responses and resistance to Edwardsiella tarda challenge in Japanese flounder (Paralichthys olivaceus, Temminck and Schlegel)

A feeding experiment was conducted to investigate the effects of high dietary intake of vitamin E (supplied as dl ‐α‐tocopheryl acetate) and n‐3 highly unsaturated fatty acid (n‐3 HUFA) on the non‐specific immune response and disease resistance in Japanese flounder Paralichthys olivaceus. Nine practical diets were formulated to contain one of three levels of vitamin E namely, 0, 80 or 200 mg kg^?1 (the total α‐tocopherol contents in the diets were 21, 97 and 213 mg kg^?1 based on analysis), and at each vitamin E level with one of three n‐3 HUFA levels i.e. 0.5%, 1.5% or 2.0%. Each diet was randomly assigned to triplicate groups of Japanese flounder (initial body weight: 40.5±1.0 g, mean±SD) in a re‐circulation rearing system. Fish were fed twice daily to apparent satiation at 07:00 and 18:00 hours for 12 weeks. During the experimental period, water temperature was maintained at 18±1°C, salinity 31–35 g L^?1, and pH 7.8–8.2. Dissolved oxygen was not less than 6 mg L^?1, and there were negligible levels of free ammonia and nitrite. The results showed that the increase in dietary n‐3 HUFA from 0.5% to 1.0% significantly decreased muscle α‐tocopherol contents in fish‐fed diets with 21 and 97 mg α‐tocopherol kg^?1 diet (P<0.05). In 1.0% HUFA groups, alternative complement pathway activity (ACH₅₀) of fish fed the diet containing the 213 mg α‐tocopherol kg^?1 diet was significantly higher than noted for fish fed the diet containing 97 mg α‐tocopherol kg^?1 diet (P<0.05). Fish fed the diet with 213 mg α‐tocopherol kg^?1 and 2.0% n‐3 HUFA had the highest lysozyme activity (131.7 U mL^?1) among all the dietary treatments. Fish fed the diets containing 97 and 213 mg α‐tocopherol kg^?1 with 1.0% n‐3 HUFA had significantly higher respiratory burst activity than those fed the diets containing 21 mg α‐tocopherol kg^?1 with 0.5 and 1.0% n‐3 HUFA (P<0.05). In the disease resistance experiment, high intake of dietary vitamin E with 213 mg α‐tocopherol kg^?1 significantly decreased cumulative mortality and delayed the days to first mortality after a 7‐day Edwardsiella tarda challenge (P<0.05). In addition, under the experimental conditions, dietary vitamin E and n‐3 HUFA had a synergistic effect on the non‐specific immune responses and disease resistance in Japanese flounder (P<0.05).     

Effect of dietary bovine lactoferrin on growth,haematology and non‐specific immune response in rainbow trout (Oncorhynchus mykiss)

The aim of this study was to investigate the effect of dietary lactoferrin (Lf) on growth, haematology and non‐specific immune response of rainbow trout Oncorhynchus mykiss (Walbaum). Fish were fed an experimental diet containing 0 (as control), 50, 100, 200 and 400 mg Lf kg^?1 diet twice daily for 8 weeks and sampled at 2, 4, 6 (immune function and growth), and 8 weeks (immune function, haematology and growth). Statistical analyses revealed no significant effects of dietary Lf on growth performance (specific growth rate, weight gain, feed conversion ratio, feed intake and condition factor) or haematological parameters (red and white blood cell count, haemoglobin, haematocrit, serum iron and total iron binding capacity [TIBC]). Among the serum non‐specific immune parameters, lysozyme activity increased significantly in fish fed 100, 200, or 400 mg Lf kg^?1 feed for 8 weeks, whereas haemolytic complement activity increased in fish fed 100 and 400 mg Lf kg^?1 diet after 6 weeks. The antiprotease activity increased in groups fed 100, 200 or 400 mg Lf kg^?1 diet after 8 weeks. However, no significant effect was observed on serum peroxidase level. It can be concluded that feeding of rainbow trout on the diet supplemented with 100 mg kg^?1 or higher for 8 weeks enhances the non‐specific immune response.     

Effects of dietary protein and lipid levels on growth,feed utilization and body composition in red‐tailed catfish juveniles (Hemibagrus wyckioides,Chaux & Fang 1949)

A feeding trial was conducted in a recycling water system during 10 weeks to determine the optimal protein to lipid ratio in Asian red‐tailed catfish (Hemibagrus wyckioides). Six diets of two protein levels (390 and 440 g kg^?1) with three lipid levels (60, 90 and 120 g kg^?1) were formulated. Fish (1.96 g) were fed six diets with four replicates to apparent satiation at a stocking density of 50 fish per tank (500 L). Faeces were collected in cultured tanks at the end of the feeding trial for digestibility measurement. Significantly, improved growth performances (P < 0.01) and higher feed utilization (P < 0.001) were observed in fish fed with higher lipid diets. However, higher protein diets did not significantly improve fish growth but they reduced FCR (P < 0.001) and protein efficiency ratio (P < 0.01). Higher lipid diets also resulted in significantly increased adipose‐somatic index, carcass fat and reduced moisture of the fish. The study revealed the protein sparing effect of dietary lipid in the catfish and highest growth performance was found by fish fed 390 g kg^?1 protein and 120 g kg^?1 lipid diet with P/E ratio of 20.48 mg protein kJ^?1. DP/DE ratio for maximal growth rate in diets was 21.48 mg protein kJ^?1.     

Haemato‐immunological response of Labeo rohita (Hamilton) fingerlings fed leaf extracts and challenged by Aeromonas hydrophila

A 35 days feeding trial was conducted to assess the haemato‐immunological response of Labeo rohita fingerlings fed ethanolic leaf extracts of Psidium guajava and Mangifera indica, and infected with Aeromonas hydrophila. Six iso‐nitrogenous (354.6–361.6 g kg^?1) purified diets were prepared with graded level of leaf extracts viz., control (basal feed without any extract); TG‐5 (5 g kg^?1 guava extract); TG‐10 (10 g kg^?1 guava extract); TM‐5 (5 g kg^?1 mango extract); TM‐10 (10 g kg^?1 mango extract); and TGM (5 g kg^?1 guava extract +5 g kg^?1 mango extract). Haematological, immunological, biochemical, along with antioxidant enzyme activities were examined after a 35 day‐feeding trial and following a 7 day challenge with A. hydrophila. The haemoglobin, total leucocyte and erythrocyte counts, respiratory burst activity, lysozyme, total protein, albumin and globulin contents increased significantly (P < 0.05) in leaf extracts fed groups compared with the control in pre‐ and post‐challenge conditions. A significant (P < 0.05) decrease was observed in SOD (superoxide dismutase) and catalase activities of the treatment groups compared with the higher value in control. The trends in mortality indicated that groups of fish showing significantly elevated haemato‐immunological responses had the lowest mortality following challenge with A. hydrophila. The results showed that extracts of P. guajava and M. indica appear to be potential immunostimulant at an inclusion level of 5 g kg^?1 in the diet of rohu. But, mixing of both the extract at similar level did not show any synergistic effect, which needs to be tested at its lower level of inclusion.     

The effect of dietary methionine on growth,antioxidant capacity,innate immune response and disease resistance of juvenile yellow catfish (Pelteobagrus fulvidraco)

An 11‐week feeding trial was conducted to evaluate the effect of dietary methionine on the growth, antioxidant status, innate immune response and disease resistance to Aeromonas hydrophila of juvenile yellow catfish. Six isonitrogenous and isolipidic practical diets were formulated to contain different graded methionine levels ranging from 6.1 to 16.4 g kg^?1 of dry weight. The results indicated that growth performance and feed utilization were significantly influenced by the dietary methionine levels; fish fed the diet containing 6.1 g kg^?1 methionine level had lower specific growth rate, percentage weight gain (PWG), feed efficiency and protein efficiency ratio than those fed the other diets (P < 0.05). Fish fed the diet containing 16.4 g kg^?1 methionine level had lowest protein contents in whole body and muscle among all treatments. Triacylglycerols, cholesterol, aspartate aminotransferase, alanine aminotransferase and haemoglobin (Hb) in plasma or whole blood were significantly affected by dietary methionine levels. Fish fed the diet containing 6.1 g kg^?1 methionine level had higher superoxide dismutase, glutathione peroxidase activities and malondialdehyde values than those fed other diets. Fish fed diets containing 9.7 and 11.8 g kg^?1 methionine levels had higher lysozyme activity, total immune globulin, phagocytic activity and respiratory burst than those fed other diets. The lowest survival after A. hydrophila challenge was observed in fish fed a diet containing 6.1 g kg^?1 methionine. Quadratic regression analysis of PWG against dietary methionine levels indicated that the optimal dietary methionine requirement for the maximum growth of juvenile yellow catfish was estimated to be 11.5 g kg^?1 of the diet in the presence of 4.0 g kg^?1 cystine (corresponding to 23.5 g kg^?1 of dietary protein on a dry weight basis).     

Requirements of vitamin C (l-ascorbyl-2-sulphate and l-ascorbyl-2-polyphosphate) and its effects on non-specific immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to determine two vitamin C derivatives, l ‐ascorbyl‐2‐sulphate (C2S) and l ‐ascorbyl‐2‐polyphosphate (C2PP), to satisfy vitamin C requirement and to test their effects on the non‐specific humoral immune responses of juvenile grouper, Epinephelus malabaricus. C2S and C2PP were each supplemented at 20, 50, 80, 150, 250 and 400 mg kg^?1 diet in the semi‐purified basal diet providing of 7, 16, 28, 55, 86, 142 mg ascorbic acid (AA) equivalent of C2S kg^?1 diet and 4, 9, 15, 31, 49, 75 mg AA equivalent of C2PP kg^?1 diet, respectively. Basal diet without AA supplemented was included as a control. Each diet was fed to triplicate groups of grouper (mean initial weight: 6.69 ± 0.07 g). Fish fed diets with ≥28 mg AA equivalent of C2S or ≥4 mg AA equivalent of C2PP kg^?1 had significantly (P < 0.05) greater weight gain (WG) than fish fed the unsupplemented control diet. Liver AA concentrations were higher in fish fed diets with ≥16 mg AA equivalent of C2S or ≥9 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Alternative pathway of complement activation (ACP) was higher in fish fed diets with ≥55 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Lysozyme activity was higher in fish fed ≥86 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Analysis by broken‐line regression of WG indicated that the adequate dietary vitamin C concentration from each vitamin C derivative in growing grouper is 46.2 mg AA equivalent of C2S kg^?1 diet and 17.8 mg AA equivalent of C2PP kg^?1 diet, and it also indicated that C2S is approximately 39% as effective as C2PP in meeting the vitamin C requirement for grouper. The data suggest that both C2S and C2PP supplementation support non‐specific immune responses of grouper.     

Effects of dietary vitamin C on immunity, growth and survival of Indian major carp Labeo rohita, fingerlings

An 8‐week feeding trial was conducted to evaluate the efficacy of dietary l ‐ascorbic acid (AA) dosages on immunity, growth and survival of the fingerlings of Labeo rohita. AA was supplemented at 0, 100, 200 and 500 mg kg⁻¹ basal diets. Each diet was fed for 8 weeks to duplicate groups of fingerlings of rohu with initial body weight of 35 ± 5 g. Selected biochemical and haematological analyses were evaluated every second week. At the end of the feeding trial of 56 days, fish were divided into two subgroups under each major treatment group for challenge with two pathogens, Aeromonas hydrophila and Edwardsiella tarda. The mortality (%) and agglutinating antibody titre were recorded on 28th day postchallenge. Non‐specific immune parameters like superoxide anion production, lysozyme activity and complement activity increased independently of the levels of AA in the diet. Whereas phagocytic ratio, phagocytic indices, lymphokine production indices and serum bactericidal activity were better in the group of fish fed with 200 mg AA kg⁻¹ feed for 42 days. However, growth parameters such as specific growth rate, feed conversion ratio and survival were better in the group of fish fed with the highest level of AA through feed for 56 days. These data indicated that elevated levels of dietary AA could be recommended for optimum immunity, growth and survival of fingerlings of L. rohita.     

Effectiveness of l‐ascorbyl‐2‐monophosphate Na/Ca as a vitamin C source for yellowtail Seriola quinqueradiata juveniles

A feeding experiment was conducted to examine the efficacy of a vitamin C (L‐ascorbic acid, AsA) derivative, l ‐ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) by comparing to l ‐ascorbyl‐2‐monophosphate‐Mg (AMP‐Mg) in terms of growth, AsA content in tissues, and hematology in yellowtail Seriola quinqueradiata juvenile (1.10 ± 0.01 g). Furthermore, a stress resistance test for the species fed AMP‐Na/Ca was also conducted. In experiment 1, 11 test pellet diets with different levels of AMP‐Na/Ca (0, 12, 43, 88, 440 and 881 mg AsA kg^?1diet) and AMP‐Mg (0, 16, 52, 106, 595 and 1164 mg AsA kg^?1 diet) were formulated and fed to the yellowtail three times per day for 50 days. In both the vitamin C sources, the survival rates of the fish fed diets without supplemental AsA were lower than 50% at day 20, and more than 50% mortality occurred in fish that fed the diets containing 12 or 16 mg AsA kg^?1 after day 30. However, no significant differences were detected in survival and growth among fish that fed the diets containing more than 43 mg AsA kg^?1. Liver and brain AsA concentrations generally increased with increasing dietary AsA level in both sources. In experiment 2, test diets were formulated to contain 43, 88, 440 and 881 mg AsA kg^?1 using AMP‐Na/Ca, and after 60 days of feeding, yellowtail juveniles were subjected to low salinity and air exposure stress. The fish that received diets with 440 mg AsA kg^?1 showed significantly higher tolerance to low salinity stress and higher survival rate in air exposure stress than those of other groups. The present study demonstrated that yellowtail juveniles could utilize AMP‐Na/Ca as an AsA source like AMP‐Mg, and that supplementation of 43–52 mg AsA kg^?1 diet was optimum for normal growth. However, this study showed that dietary inclusion of 440 mg AsA kg^?1 would be necessary to improve stress resistance of this species.     

Effects of dietary squid viscera meal on growth and cadmium accumulation in tissues of Japanese seabass, Lateolabrax japonicus (Cuvier 1828)

Cadmium (Cd) is a toxic environmental pollutant with a long biological half‐life and can produce both hepatic and renal injuries in mammals and fish. Squid viscera meal (SVM), an effective attractant for aquatic animals, is widely used as an ingredient in aquafeeds. However, SVM is rich in Cd and its complexes. A study was conducted to evaluate the effects of dietary SVM on the growth and Cd deposition in tissues of Japanese seabass, Lateolabrax japonicus. Three practical diets were formulated to contain 0, 50 and 100 g SVM kg^?1 diet, respectively, containing 0.21, 7.26 and 12.08 mg Cd kg^?1 diet. Each diet was randomly assigned to triplicate groups of 80 Japanese seabass (mean initial weight, 10.89±0.21 g) in floating sea cages (1.5 × 1.5 × 2.0 m). Fish were fed twice daily (06:30 and 16:30 hours) to satiation for 8 weeks. The results showed that there were no significant differences in fish survival among three dietary treatments, but significant higher specific growth rates (SGR) were observed in the fish fed diets with 50 or 100 g SVM kg^?1 diet than that from the control group (P<0.05). The Cd concentrations in the kidney, liver and gill were found in a decreasing order at each treatment, and positively correlated with dietary Cd levels. Fish fed diets with 50 and 100 g SVM kg^?1 diet had significantly higher Cd accumulations in the kidney (3.25, 5.85 mg kg^?1), liver (0.76, 1.26 mg kg^?1) and gill (0.42, 0.58 mg kg^?1) compared with the control group (0.82, 0.34 and 0.32 mg kg^?1 respectively) (P<0.05). The Cd concentration in fish muscle; however, was undetectable in any treatment. Therefore, based on these results, accumulation of Cd in edible tissue (muscle) of farmed Japanese seabass is not a food safety issue. However, long‐term feeding of diets with SVM may result in accumulation of Cd in the kidneys, liver and gills of fish.