Terminalia arjuna Bark Powder as a Potential Immunomodulator in Labeo rohita: Enhanced Hematological, Adaptive, and Humoral Responses against Bacterial Pathogens and Concordant Liver Histomorphology.

This study investigated the dietary immunomodulatory effects of Terminalia arjuna bark powder (TABP) in Labeo rohita, a freshwater fish model. Four iso-nitrogenous and iso-caloric diets containing graded levels of TABP (0, 1, 10, and 15 g/kg were fed to fish for 90 days, followed by a 10 day challenge with pathogenic bacteria Aeromonas hydrophila and Edwardsiella tarda. An integrated biomarker response (IBR) approach assessed the impact of TABP on hematological, adaptive, and humoral immune parameters, along with liver histomorphology. Dietary TABP at 10 g/kg significantly enhanced (p < 0.05) hematological indices (hemoglobin, red blood cell count, hematocrit), specific immune parameters (lysosomal enzyme activity, phagocytosis, respiratory burst), and non-specific immune parameters (serum lysozyme, alternative complement activity), and exhibited improvements in liver architecture consistent with the enhanced immune response. Broken line regression analysis showed 11.5 g/kg to be an optimum dose. However, at 15 g/kg, a compromised trend was observed in some parameters. These findings suggest an optimal dosage range for TABP's immunomodulatory effects. The study highlights the potential of TABP as a natural immunomodulator in fish aquaculture. The improved immune response and concomitant liver health observed in Labeo rohita opens avenues for further research on TABP's applicability in animal health, using fish as a model organism. Additionally, the IBR approach proved effective in evaluating TABP's immunomodulatory properties, paving the way for similar studies on other natural products in aquaculture.

Effect of sulfate on the osmoregulatory and physio-biochemical responses of GIFT (Oreochromis niloticus) juveniles reared in potassium-deficient medium saline waters.

The inland saline waters were continuously observed to have low potassium concentrations compared to their seawater counterpart of the same salinity. We hypothesize that the toxic effect of sulfate may manifest in low potassium saline (LPSW) waters compared to brackish water of the same salinity. Thus, LC50 trials were performed in GIFT (genetically improved farmed tilapia) fry (0.5 ± 0.02 g) to determine the acute sulfate toxicity in freshwater (FW, 0.5 g L-1), artificial seawater (ASW, 10 g L-1), and LPSW (10 g L-1). The median lethal concentrations (96h LC50) of sulfate ion in FW, LPSW, and ASW for the GIFT were 5.30 g L-1, 2.56 g L-1, and 2.98 g L-1, respectively. A second experiment was conducted for 21 days, exposing fish to a sub-lethal level of sulfate ion (SO42-) concentration (1000 mg L-1, one-fifth of FW LC50) with different types of waters (FW, freshwater, 0.5 g L-1; ASW, artificial seawater, 10 g L-1; LPSW, low potassium saline water, 10 g L-1) with and without sulfate inclusion to constitute the treatments as follows, (FW, FW + SO4, ASW, ASW + SO4, LPSW, LPSW + SO4). The effect of sulfate on GIFT reared in sulfate-rich potassium-deficient medium saline water was evaluated by focusing on the hematological adjustments, stress-induced oxidative damage, and osmoregulatory imbalances. The survival was not altered due to the sulfate concentration and K+ deficiency; however, there were significant changes in branchial NKA (Na+/K+-ATPase) activity and osmolality. The increase in NKA was highest in LPSW treatment, suggesting that internal ionic imbalance was triggered due to an interactive effect of sulfate and K+ deficiency. The cortisol levels showed a pronounced increase due to sulfate inclusion irrespective of K+ deficiency. The antioxidant enzymes, i.e., SOD (superoxide dismutase), catalase, GST (glutathione-S-transferase), and GPX (glutathione peroxidase), reflected a similar pattern of increment in the gills and liver of the LPSW + SO4 groups, suggesting a poor antioxidant status of the exposed group. The hepatic peroxidation status, i.e. TBARS (thiobarbituric acid reactive substances), and the peroxide values were enhanced due to both K+ deficiency and sulfate inclusion, suggesting a possible lipid peroxidation in the liver due to handling the excess sulfate anion concentration. The hematological parameters, including haemoglobin, total erythrocyte count, and hematocrit level, reduced significantly in the LPSW + SO4 group, indicating a reduced blood oxygen capacity due to the sulfate exposure and water potassium deficiency. The hepatic acetylcholine esterase activity was suppressed in all the treatments with sulfate inclusion, while the highest suppression was observed in the LPSW + SO4 group. Thus, it is concluded that sulfate-induced physiological imbalances manifest more in potassium-deficient water, indicating that environmental sulfate is more detrimental to inland saline water than freshwater or brackish water of the same salinity.

Sugarcane bagasse biochar enhances the growth parameters, haematological parameters, and enzyme activities of genetically improved farmed tilapia (GIFT) reared in inland saline water.

In the present scenario, waste management, especially agro-waste, is one of the major challenges. India is an agrarian country and the economy depends on agriculture and a huge amount of agro-wastes are generated. In this study, biochar was prepared from paddy straw and sugarcane bagasse and was used in the feed of genetically improved farm tilapia (GIFT) at 0.5% (w/w) basis to study its effect on growth and haematological parameters. An experiment was carried out in triplicate in 500-l capacity FRP tanks comprising two treatments and one control. Tanks were filled with inland saline soil to maintain 25-cm soil's bed and water with 12 ppt salinity. A total of 22 fish having an average length of 5.14 ± 0.07 cm and weight of 4.8 ± 0.05 g were stocked in each tank; the fish were fed at apparent satiation level, twice daily for 45 days. It was observed that growth parameters like SGR% (specific growth rate), weight gain (WG%), and feed conversion ratio (FCR) were significantly improved (p < 0.05) than the control. Biochar-enriched feed also significantly improved the haematological parameters like red blood cell (RBC), white blood cell (WBC), haemoglobin (Hb) haematocrit % (HCT), mean cell haemoglobin concentration (MCH), and MCV (mean cell volume) in treatment groups than the control. Feeding biochar as feed additives was also found to decrease catalase (CAT) and enhanced amylase and lipase activities in treatments as compared to control (p < 0.05). It can be concluded that the application of biochar as a feed additive enhanced the growth and overall health of the fish, and it can enhance fish production. However, biochar from sugarcane bagasse was found to be more effective than the paddy straw biochar in the diet of tilapia (GIFT).

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.

Biometric Indices, Physio-Metabolic Responses and Carcass Quality in Rohu (Labeo rohita) during Feed Deprivation.

Understanding changes in biometric indices and metabolism in fish exposed to feed deprivation may be useful in aquaculture. The present study elucidates the effect of feed deprivation on physio-biochemical responses, such as changes in biometric indices, nutrient mobilization patterns, and enzyme activities in rohu (Labeo rohita). Experimental fish (av. wt. 3.41 ± 0.07 g) were deprived of feed and sampled at intervals of 0, 15, 30, 45, and 60 days to measure weight, length, body composition, and the activities of enzymes involved in digestion, metabolism, and antioxidation. A decrease in body weight, condition factor (CF), hepatosomatic index (HSI), and gastrosomatic index (GSI) was observed during the initial stage of feed deprivation (15 to 30 days) but remained unchanged thereafter. The total carbohydrate and lipid content also decreased rapidly up until 30 days, then stabilized. However, the reduction in tissue protein content (% wet weight) continued gradually with the duration of feed deprivation from 12.85 ± 0.36 at 0 days to 10.04 ± 0.67 at 15 days, 8.79 ± 0.59 at 30 days, 6.95 ± 0.69 at 45 days, and 6.16 ± 0.8 at 60 days, which was lower, compared to the other two body constituents. Amylase, protease. and lipase activities significantly reduced up until 30 days, but then stabilized. Although G6PDH enzyme activity decreased, gluconeogenic (LDH, AST, and ALT) and antioxidative (SOD and catalase) enzyme activities increased during initial feed deprivation (up to 30 days). A tissue-specific difference in amino acid metabolism with a major role of ALT in liver and AST in muscle was observed. This study revealed that rohu fingerlings adapted well to feed deprivation up until 30 days, beyond which there was an overall deterioration in the metabolic functions.

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.

Effect of Dietary Lipid Level on Growth Performance, Body Composition, and Physiometabolic Responses of Genetically Improved Farmed Tilapia (GIFT) Juveniles Reared in Inland Ground Saline Water.

A 60-day feeding trial was carried out to determine the effect of dietary lipid levels on growth and physiometabolic responses to optimize the dietary lipid requirement for maximizing the growth of Genetically Improved FarmedTilapia (GIFT) juveniles reared in inland ground saline water (IGSW) of medium salinity (15 ppt). Formulation and preparation of seven heterocaloric (389.56-449.02 Kcal digestible energy/100 g), heterolipidic (40-160 g/kg), and isonitrogenous (410 g/kg crude protein) purified diets were done for conducting the feeding trial. Random distribution of 315 acclimatized fish (mean weight 1.90 ± 0.01 g) was made in seven experimental groups such as CL4 (40 g/kg lipid), CL6 (60 g/kg lipid), CL8 (80 g/kg lipid), CL10 (100 g/kg lipid), CL12 (120 g/kg lipid), CP14 (140 g/kg lipid), and CL16 (160 g/kg lipid) with 15 fish per triplicate tank (fish density, 0.21 kg/m3). Respective diets were used for feeding the fish at satiation level three times daily. Results indicated that weight gain percentage (WG%), specific growth rate (SGR), protein efficiency ratio, and protease activity significantly increased up to 100 g lipid/kg fed group, and then the values significantly decreased. Muscle ribonucleic acid (RNA) content and lipase activity were highest in 120 g/kg lipid-fed group. RNA/DNA (deoxyribonucleic acid) and serum high-density lipoproteins levels of 100 g/kg lipid-fed group were significantly higher than 140, and 160 g/kg lipid-fed groups. The lowest feed conversion ratio was found in the 100 g/kg lipid-fed group. The amylase activity was significantly higher in 40 and 60 g lipid/kg fed groups. The whole-body lipid level was increased with increasing the dietary lipid levels, whereas, there was no significant difference in whole-body moisture, crude protein, and crude ash contents of all groups. Highest serum glucose, total protein and albumin, and albumin to globulin ratio and lowest low-density lipoproteins level were found in 140 and 160 g/kg lipid-fed groups. Serum osmolality and osmoregulatory capacity did not vary significantly, whereas carnitine palmitoyltransferase-I and glucose-6-phosphate dehydrogenase showed an increased and decreased trend, respectively, with the increasing dietary lipid levels. According to second-order polynomial regression analysis based on WG% and SGR, the optimum dietary lipid for GIFT juveniles in IGSW of 15 ppt salinity was found to be 99.1 and 100.1 g/kg, respectively.

Immunogenic Effects of Dietary Terminalia arjuna Bark Powder in Labeo rohita, a Fish Model: Elucidated by an Integrated Biomarker Response Approach.

Utilizing agro-industrial waste and herbal products to create a circular bioeconomy is becoming increasingly popular. Terminalia arjuna is a significant ethnomedicinal plant that has not yet been exploited in animal feed. In the present study, nutritional Terminalia arjuna bark powder-based fish feed was created and supplied to a candidate fish species Labeo rohita at varied levels: 0% (0 g/kg), 0.5% (5 g/kg), 1% (10 g/kg), and 1.5% (15 g/kg). These treatment groups are denoted as CT, T1, T2, and T3, respectively. Utilizing a contemporary comprehensive biomarker response strategy, the study clarified the genomic influence of dietary herb inclusion. In response to bacterial infection, the immunogenic genes, STAT 1 (signal transducer and activator of transcription 1), ISG 15 (interferon stimulating gene), and Mx "myxovirus resistance gene", were shown to be elevated. The results of densitometry demonstrated a dose-dependent increase in STAT 1 and ISG 15, with Mx exhibiting maximal values at 1 g/kg TABP (Terminalia arjuna bark powder-based feed). This is the first study to identify TABP as an immunomodulator in fish and established the IBR (Integrated Bio-marker Response) as a reliable marker in evaluating the impact of multiple drivers in a holistic manner. Thus, the present study cleared the path for TABP to be utilized as an effective feed additive which enhances the specific adaptive immune system of the fish for the production of the Green fish product for a sustainable circular bioeconomy.

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.

Scope of Archaea in Fish Feed: a New Chapter in Aquafeed Probiotics?

The outbreak of diseases leading to substantial loss is a major bottleneck in aquaculture. Over the last decades, the concept of using feed probiotics was more in focus to address the growth and health of cultivable aquatic organisms. The objective of this review is to provide an overview of the distinct functionality of archaea from conventional probiotics in nutrient utilization, specific caloric contribution, evading immune response and processing thermal resistance. The prime limitation of conventional probiotics is the viability of desired microbes under harsh feed processing conditions. To overcome the constraints of commercial probiotics pertaining to incompatibility towards industrial processing procedure, a super microbe, archaea, appears to be a potential alternative approach in aquaculture. The peculiarity of the archaeal cell wall provides them with heat stability and rigidity under industrial processing conditions. Besides, archaea being one of the gut microbial communities participates in various health-oriented biological functions in animals. Thus, the current review devoted that administration of archaea in aquafeed could be a promising strategy in aquaculture. Archaea may be used as a potential probiotic with the possible modes of functions and advantages over conventional probiotics in aquafeed preparation. The present review also provides the challenges associated with the use of archaea for aquaculture and a brief outline of the patents on archaea to highlight the various use of archaea in different sectors.

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.

Pseudobranch mimics gill in expressing Na+K+-ATPase 1 α-subunit and carbonic anhydrase in concert with H+-ATPase in adult hilsa (Tenualosa ilisha) during river migration.

In hilsa (Tenualosa ilisha), pseudobranch comprises a row of parallel filaments bear numerous leaf-like lamellae arranged on both sides throughout its length. The purpose of this study was to elucidate involvement of pseudobranchial Na+, K+-ATPase (NKA) 1 α-subunit, and carbonic anhydrase (CA) in concert with H+-ATPase (HAT) compared to their branchial counterparts in freshwater acclimation of hilsa during spawning migration from off-shore of the Bay of Bengal to the Bhagirathi-Hooghly zones of the Ganga river system in India. Adult hilsa fish were collected from seawater (SW), freshwater 1 (FW1), and freshwater 2 (FW2) locations, where the salinity level was 26-28‰, 1-5‰, and 0-0.04‰, respectively. Hilsa migrating through freshwater showed a consistent decrease in the plasma osmolality, sodium (Na+) and chloride (Cl-) ion levels indicates unstable ionic homeostasis. The mRNA expression and activity of NKA 1 α-subunit in pseudobranch as well as in true gills declined with the migration to upstream locations. The pseudobranchial CA activity almost mirrors its branchial counterpart most notably while hilsa entered the freshwater zone, in the upstream river suggesting its diverse role in hypo-osmotic regulatory acclimation. Nevertheless, the H+-ATPase activity of both the tissues increased with the freshwater entry and remained similar during up-river movement into the freshwater environment. The results confirm that the pseudobranchial NKA 1 α-subunit mRNA expression and activity mimic its branchial counterpart in the process of ionoregulatory acclimation during migration through salt barriers. Also, the increase in the activities of pseudobranchial and branchial CA in concert with H+-ATPase (HAT) during freshwater acclimation of hilsa suggests their critical involvement in ion uptake.

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.

Substitution of soybean meal with fermented Jatropha kernel meal: effect on growth performance, body composition, and metabolic enzyme activity of Labeo rohita.

A 60-day feeding trial was conducted to evaluate the nutritional potential of fermented Jatropha kernel meal (FJKM) in the diets of rohu, Labeo rohita, fingerlings. Four iso-nitrogenous (336 g kg-1) and iso-energetic (20 MJ kg-1) diets containing 0, 100, 200, and 300 g kg-1 FJKM in replacement of 0, 33.3, 66.7, and 100% soybean meal protein, respectively, were prepared and designated as control, T1, T2, and T3, respectively. A total of 180 acclimatized fish (average weight 6.05 ± 0.04 g) were equally distributed into four experimental groups in triplicates, and were fed their respective diets to satiation twice daily at 10:00 h and 18:00 h. Higher percent weight gain (%WG); specific growth rate (SGR); protein efficiency ratio (PER); protease, aspartate amino transferase (AST), and alanine amino transferase (ALT) activities; and lower feed conversion ratio (FCR) were recorded in T2 and T3 groups compared to other groups. Whereas hepatosomatic index (HSI), intestinal somatic index (ISI), and amylase, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and catalase activities among the various dietary groups did not vary significantly (P > 0.05). Whole body composition of fish analyzed at the end of the feeding trial exhibited significantly (P < 0.05) higher ether extract and lower crude protein in the control group compared to the FJKM-fed groups. These results indicated that rohu fingerlings can efficiently utilize FJKM without any detrimental effect on growth performance, nutrient utilization, and metabolic response.

Effects of fluconazole based medicated feed on haemato-immunological responses and resistance of Labeo rohita against Saprolegnia parasitica.

Fluconazole (FLZ) is a new azole antifungal drug having no earlier record of its utilization for the treatment of fish diseases. A 55-days experiment was carried out to delineate the role of FLZ based medicated feed on haemato-immunological responses and prevention of fungal infection in Labeo rohita fingerlings. Three hundred and sixty fingerlings were randomly distributed into four experimental groups in triplicates. Four iso-caloric and iso-nitrogenous purified medicated feeds were prepared with 0, 100, 200 and 300 mg FLZ 100 g-1 feed. Haemato-immunological parameters like erythrocyte counts (EC), haemoglobulin (Hb), leucocyte counts (LC), packed cell volume (PCV), mean corpuscular haemoglobin (MCH), mean corpuscular volume (MCV), mean corpuscular haemoglobin concentration (MCHC), nitro-blue tetrazolium (NBT), albumin, globulin, total plasma protein were studied. The present study revealed that the continuous feeding of FLZ based feed for 15-30 days significantly (P < 0.05) increases the erythropoiesis, heme-synthesis, as well as the leucocytosis in rohu. However, all the doses exhibited equal protection from the infectious S. parasitica after 45 days of continuous feeding.

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.