Complimentary effect of exogenous enzymes, essential amino acids and essential fatty acids supplemented de-oiled rice bran (DORB) based diets on hematology, liver and intestinal histoarchitecture in Labeo rohita.

A 60-day feeding trial was conducted to study the hematology, liver, and intestinal histoarchitecture of Labeo rohita fed with a combination of exogenous enzymes, essential amino acids, and essential fatty acids to DORB (De-oiled rice bran) based diets. Three treatments viz., T1 [DORB + phytase and xylanase (0.01% each)], T2 [DORB + phytase (0.01%) + xylanase (0.01%) + L-lysine(1.4%) + L-methionine (0.4%) + EPA and DHA (0.5%)] and T3 [DORB + phytase (0.01%), xylanase and cellulase (0.075%) + L-lysine (1.4%) +L-methionine (0.4%) + EPA and DHA (0.5%)] were used in the present study. Serum total protein, albumin content and A/G ratio varied significantly (p < 0.05) among groups. Globulin content did not vary significantly among groups (p ≥ 0.05). The Hb content, RBC and MCV count varied significantly (p < 0.05) whereas MCH, MCHC content, WBC and lymphocyte count did not vary significantly among groups (p > 0.05). The liver and intestine examination revealed no visible alteration and showed normal histo-architecture. Based on the finding it is concluded that DORB supplemented with exogenous enzymes, essential amino acids and essential fatty acids with phytase (0.01%), xylanase and cellulase (0.075%), L-lysine (1.4%), DL-methionine (0.4%) and EPA and DHA (0.5%) improves the health of L. rohita.

Taurine and/or inorganic potassium as dietary osmolyte counter the stress and enhance the growth of GIFT reared in ion imbalanced low saline water.

The effects of dietary osmolytes for alleviating osmotic stress and enhancing growth are not well elucidated in fish reared in inland saline water. The present study evaluated the effects of dietary taurine or potassium (K+) individually or in combination on growth, ionic homeostasis, and stress response of GIFT tilapia reared in potassium deficient low saline water (PDLSW, 10 ppt salinity) mimicking inland saline water. Isonitrogenous and isoenergetic diets supplemented with five potassium concentrations (0, 0.3, 0.45, 0.6 and 0.75 %), two taurine (T) concentrations (0.5 and 1.0 %) and two combinations of both (K+ 0.1 % + T 0.5 % and K+ 0.2 % + T 0.5 %) were fed to GIFT juveniles (4.4 ± 0.02 g body weight) and reared in PDLSW for 45 days. The fish fed on the diet fortifying with K+ 0.2 % + T 0.5 % showed the highest growth performance among the controls and other treatment groups. Dietary supplementation had no effects on PDLSW induced increase in osmoregulatory endpoints. The optimum dietary potassium requirement of GIFT reared in PDLSW was 0.57 and 0.599 g/100 g diet. Dietary K+ down-regulated the PDLSW induced expression of NKAa1, AQP1, and ClC2, whereas inhibited taurine-induced up-regulation of AQP1 and CLC2, which is the first report in tilapia. In addition, dietary K+ and taurine modulated antioxidant and metabolic enzyme activities for easing stress and balancing energy requirements. Thus, blending of potassium (0.2 %) and taurine (0.5 %) in the diet appears best to mitigate stress and enhance GIFT growth reared in inland saline water.

Dietary arginine attenuates hypoxia- induced HIF expression, metabolic responses and oxidative stress in Indian Major Carp, Cirrhinus mrigala.

Hypoxia is a common stressor in aquaculture systems, which causes severe physiological disturbances, ultimately leading to mortality or reduced productivity. Arginine, as a precursor of NO, has a role in enhancing oxygen delivery. Thus, an experiment was conducted to evaluate the effect of dietary arginine (Arg) in Cirrhinus mrigala exposed to hypoxia. The fish were fed with different levels of arginine for 60 days and exposed for 72 h to a sublethal level of hypoxia (0.50 ± 0.16 mg/L dissolved oxygen [DO]). The six treatment groups with three replicates were N0 (0% Arg + Normoxia), H0 (0% Arg + Hypoxia), N0.7 (0.70% Arg + Normoxia), H0.7 (0.70% Arg + Hypoxia), N1.4 (1.40% Arg + Normoxia), H1.4 (1.40% Arg + Hypoxia). Eighteen experimental units with twelve animals (5.8 ± 0.18 g) each were used for the trial.The results indicated that supplementation of arginine at 0.7 and 1.4% enhanced the hypoxia tolerance time, although the high dose (1.4%) did not yield any further increments. The exposure to hypoxia up-regulated Hypoxia Inducible Factor (HIF)-1α mRNA expression and supplementation of arginine significantly decreased hypoxia induced up-regulation of HIF at 1.4%. Arginine supplementation partially or completely normalised the hypoxia induced changes in the metabolic enzymes of C. mrigala. The fish exposed to hypoxic conditions exhibited significantly higher (P < 0.05) lipid peroxidation levels than those maintained under normoxic conditions, while arginine feeding significant in reducing lipid peroxidation. Antioxidant enzyme activities were significantly higher (P < 0.05) in hypoxia-exposed carp, indicating increased oxidative stress during the hypoxic exposure, that was improved in Arg-supplemented groups. However, arginine did not modulate erythrocyte countsalthough itreduced the erythrocyte fragility. We conclude arginine supplementation is effective in ameliorating hypoxia induced metabolic alterations and improving antioxidant defences in fish.

Changes in weight gain, digestive and metabolic enzyme activities in Labeo rohita fingerlings in response to multiple stress exposure and dietary nutraceutical.

A feeding trial was conducted for 4 months to study the effect of a nutraceutical conglomerate at different levels (0, 0.1, and 0.5%) on stress-exposed (high stocking density, H and feed restriction, R) Labeo rohita fingerlings. Six isonitrogenous diets with a crude protein of 35% were prepared for the different treatments, viz. THR0, THR1, THR5, TNS0, TNS1 and TNS5. Stress significantly reduced the weight gain and feeding of 0.1% nutraceutical improved it in both stress-exposed and non-exposed groups from 3 months onwards. Two-and three-month stress-exposed groups exhibited lower (p < 0.05) protease activity, while amylase activity was significantly higher in 2-month stress-exposed groups. Higher lipase activity was found in stress-exposed groups irrespective of the duration of stress. Higher serum glutathione peroxidase (GPx), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and hepatic and muscle lactate dehydrogenase (LDH) activities were observed in the THR0 groups; however, THR1 showed a lower activity of these enzymes. Increased malate dehydrogenase (MDH) and glucose-6-phosphate dehydrogenase (G6PDH) activity was recorded in the 1st and 2nd months stress-exposed groups. However, THR1 group exhibited a reduced MDH and G6PDH activity compared to THR0 group in 1st and 2nd months. Hence, it can be concluded that the duration of multiple stress exposure adversely affected the digestive and metabolic enzymes activities and feeding of 0.1% nutraceutical conglomerates could restore the activities of digestive and metabolic enzymes in rohu fingerlings.

Improvement in the growth status and carbohydrate utilization of Labeo rohita (Hamilton, 1822) fingerlings with dietary supplementation of chromium picolinate.

A 60-day feeding experiment was conducted to evaluate the effects of dietary chromium (Cr) on carbohydrate utilization and growth performance of Labeo rohita fingerlings. Fishes were fed with four high carbohydrate (53%), isonitrogenous (crude protein 35%), and isocaloric (415 Kcal, 100 gm-1) experimental diets containing different levels of dietary chromium picolinate (Cr-Pic) viz.0, 400, 800, and 1200 µg kg-1 diet. Weight gain (WG%), specific growth rate (SGR), feed efficiency ratio (FER), and protein efficiency ratio (PER) were significantly increased at 800 µg kg-1 diet chromium supplementation (P < 0.05). Cr-Pic supplementation (800 µg kg-1) also significantly (P < 0.05) enhanced the protein: DNA ratio in muscle, while DNA: RNA and DNA: tissue ratios were significantly (P < 0.05) decreased indicating higher growth. Significantly higher amylase, protease, and lipase activities were recorded in 800 µg Cr-Pic kg-1 diet fed fishes (P < 0.05), while any of the experimental groups showing no significant (P > 0.05) change in hexokinase activity, indicating normal glycolysis in all. Furthermore, significant (P < 0.05) decrease of glucose-6-phospatase activity in 800 µg Cr-Pic kg-1 diet fed group, showcasing an evidence for protein-sparing action with Cr-Pic supplementation. Significantly (P < 0.05) higher serum insulin and liver glycogen in 800 µg Cr-Pic kg-1 diet fed fishes denote an improvement in carbohydrate metabolism. However, significantly (P < 0.05) higher ATPase and SOD activities were also observed when chromium supplementation was more than 800 µg kg-1 diet, indicating stress at higher level. The present study indicates that growth and carbohydrate utilization can significantly (P < 0.05) be improved by feeding the L. rohita fingerlings with Cr-Pic (800 µg kg-1 diet) supplemented diet in laboratory condition.

Exogenous phytase and xylanase supplementation of formulated diets for rohu (Labeo rohita): impact on haematology, histology and IGF I gene expression.

The present study aims to delineate the effect of exogenous enzyme supplementation of fermented or non-fermented de-oiled rice bran (DORB) on haematology, histology and expression of IGF I gene expression of Labeo rohita. Four test diets, namely, T1 (DORB), T2 (fermented DORB), T3 (DORB+exogenous enzyme) and T4 (fermented DORB+exogenous enzyme) were formulated and fed to the L. rohita for a period of 60 days. The test diets T3 and T4 were supplemented with 0.1 g kg-1 xylanase (16,000 U kg-1) and 0.1 g kg-1 phytase (500 U kg-1) enzymes. A total of 120 juveniles of L. rohita (average weight 5.01 ± 0.02 g) were stocked in 12 rectangular tanks with 10 fish per tank in triplicates. At the end of the experiment, haematology, histology and IGF I gene expression of the different groups were analysed. The haemoglobin (Hb) content, RBC count and WBC count of L. rohita varied significantly (p < 0.05) among different groups. T3 diet-fed group had the highest Hb content, RBC count and WBC count as compared to the other groups. The MCV content was significantly (p < 0.05) lower in T3 group. The DORB type, exogenous enzyme supplementation and their interaction have no significant (p > 0.05) effect on MCH, MCHC and lymphocyte content of the cultured fish. The group which were fed T3 diet had significantly (p < 0.05) higher IGF-I gene expression as compared to other groups. The histological examination of liver revealed no pathological alteration of this organ. Similarly, there were no pathological changes observed in intestinal tissue in any dietary treatment group. Based on the findings of the present study, it is concluded that exogenous enzyme supplementation of DORB-based diets improves the physiological status and growth performances of Labeo rohita.

Solid state fermentation of de-oiled rice bran: Effect on in vitro protein digestibility, fatty acid profile and anti-nutritional factors.

An experiment was conducted to study the effect of solid state fermentation of de-oiled rice bran (DORB) with Rhizopus oryzae on in vitro protein digestibility, anti-nutritional factors and fatty acid profile. The fermentation of DORB with Rhizopus oryzae significantly reduced the in vitro protein digestibility of DORB (p < .05). The fermentation of DORB with Rhizopus oryzae increased the saturated fatty acid (SFA) content by 46.83%, while decreased the MUFA and PUFA contents by 14.01 and 8.76%, respectively. The n-6 fatty acid content of FDORB increased by 6.19%, while n-3 fatty acid content decreased by 53.92%. The fermentation of DORB resulted in significant reduction in phytate and trypsin inhibitor activity (p < .05). Based on the present result it is concluded that the fermentation of DORB with Rhizopus oryzae improves the n-6 fatty acid profile and brings significant reduction in the phyate and trypsin inhibitor content but fails to improve the in vitro protein digestibility and hence cannot be recommended as a suitable microbe for DORB fermentation.

Comparative evaluation of fermented and non-fermented de-oiled rice bran with or without exogenous enzymes supplementation in the diet of Labeo rohita (Hamilton, 1822).

A 60-day feeding trial was conducted to study the effect of exogenous enzymes (xylanase and phytase) supplementation in the non-fermented and fermented de-oiled rice bran (DORB)-based diet of Labeo rohita. Four test diets (T1-DORB-based diet, T2-fermented DORB-based diet, T3-phytase and xylanase supplemented DORB-based diet, and T4-phytase and xylanase supplemented fermented DORB-based diet) were formulated and fed to the respective groups. Test diets T3 and T4 were supplemented with 0.01% xylanase (16,000 U kg-1) and 0.01% phytase (500 U kg-1) enzymes. One hundred twenty juveniles of L. rohita, with an average weight 5.01 ± 0.02 g, were stocked in 12 uniform size plastic rectangular tanks in triplicate with 10 fishes per tank following a completely randomized design (CRD). Exogenous enzyme supplementation to the T3 group significantly improved the growth performance of L. rohita (p < 0.05). Fermented DORB fed groups registered significantly lower growth irrespective of the supplementation of exogenous enzymes. The carcass composition (except CP %), enzyme activities (except amylase activity), globulin, and A/G ratio did not vary significantly (p > 0.05). Based on the results of the present study, it is concluded that exogenous enzyme supplementation significantly increases the growth of fish fed with DORB-based diet.

Feeding pyridoxine prevents Saprolegnia parasitica infection in fish Labeo rohita.

A 60-day experiment was carried out to delineate the role of dietary pyridoxine (DP) in Labeo rohita fingerlings in modulating immunity and prevention of fungal infection. Two hundred and seventy fingerlings were randomly distributed into three treatments in triplicates. Three iso-caloric and iso-nitrogenous purified diets were prepared with graded levels of pyridoxine. Three experimental groups were C (0.0% DP), T1 (0.01% DP) and T2 (0.02% DP). The role of dietary pyridoxine in modulating immunity and prevention of fungal infection was assessed by haemato-immunological parameters like erythrocyte counts (EC), leucocyte counts (LC), haemoglobulin (Hb), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), nitro-blue tetrazolium (NBT), phagocytic activity, albumin, globulin, total plasma protein, albumin/globulin and by challenge study with Saprolegnia parasitica, where relative percentage survival (RPS) were recorded. Hb, PCV, MCV, MCH, NBT, total plasma protein, albumin, globulin contents, lysozyme and phagocytic activity was significantly (P < 0.05) higher in DP fed group. Significantly (P < 0.05) higher RPS was recorded from T2 group fed with 0.02% DP for 45 days. Hence, DP has the capacity to stimulate nonspecific immunity and increase resistance to S. parasitica infection in L. rohita fingerlings.

Physiological changes in Labeo rohita during nitrite exposure: detoxification through dietary vitamin E.

This study investigated the effect of sub-lethal nitrite-nitrogen exposure on Labeo rohita. Fishes fed with different levels of vitamin E (VE) for 60days were exposed to nitrite for another 45days with same feeding regime. There were four treatment groups, viz., VE100-N, VE100+N, VE150+N and VE300+N. After 45days of exposure, lowest specific growth rate was observed in VE100+N and highest in VE100-N. Reno-somatic index and methaemoglobin reductase activity were significantly increased by nitrite exposure. Highest Hb and Hct were observed in VE100-N and significantly decreased upon nitrite exposure. Significant differences were observed in the activities of catalase and SOD as well as serum potassium and chloride levels among different treatments. However, serum calcium and osmolality was not significantly varied. Nitrite exposure caused marked increase in nitrite concentration in gill, liver and muscle. In liver and muscle dietary supplementation of higher amounts of VE found to reduce nitrite accumulation. It was noticed that nitrite exposure has adversely affected growth, haematological variables, ionic balance and dietary supplementation with additional amounts of VE found to overcome the adverse effects of nitrite-nitrogen. Detoxification of nitrite by methaemoglobin reductase system was enhanced by dietary supplementation of additional amounts of VE.