Molecular characterization and expression of type-I interferon gene in Labeo rohita.

Genes coding for type-I interferon (I-IFN) has been cloned from Labeo rohita, a commercially important and widely cultured fish in India and South East Asia. In the present study, full-length gene of I-IFN was amplified and sequenced. The sequence analysis revealed that I-IFN consists of 1,786 bp genomic sequence with four introns and five exons and an ORF of 546 bp encoding for a putative protein of 181 amino acids. The mature protein has a molecular weight of 18.97 kDa and consists of 158 amino acids and a signal peptide of 23 amino acids at the N terminus. The sequence carries I-IFN signature motif, one glycosylation site, two conserved cystine amino acids and other conserved amino acids. The sequence showed highest similarity to that of Cyprinus carpio (84%). In silico analysis of the rohu I-IFN protein was done using various bioinformatic tools. The constitutive expression of I-IFN gene was found to be more in spleen compared to gill and kidney in real time PCR assay. Expression of I-IFN increased about 20-fold in cultured kidney cell 2 h after induction with poly I:C and showed maximum expression at 8 h post-induction.

Beta-glucan: an ideal immunostimulant in aquaculture (a review).

The major hindrance in the development and sustainability of aquaculture industry is the occurrence of various diseases in the farming systems. Today, preventive and management measures are central concern to overcome such outbreak of diseases. Immunostimulants are considered as an effective tool for enhancing immune status of cultured organisms. Among different immunostimulants used in aquaculture practices, ß-glucan is one of the promising immunostimulant, which is a homopolysaccharide of glucose molecule linked by the glycoside bond. It forms the major constituents of cell wall of some plants, fungi, bacteria, mushroom, yeast, and seaweeds. Major attention on ß-glucan was captivated with the gain in knowledge on its receptors and the mechanism of action. The receptor present inside the animal body recognizes and binds to ß-glucan, which in turn renders the animal with high resistance and enhanced immune response. This review highlights ß-glucan as an immunostimulant, its effective dosages, and route of administration and furthermore provides an outline on role of ß-glucan in enhancing growth, survival, and protection against infectious pathogens pertaining to fishes and shellfishes. Study also summarizes the effect of ß-glucan on its receptors, recognition of proteins, immune-related enzymes, immune-related gene expression and their mechanisms of action.

Acute and chronic effects of endosulfan on the haemato-immunological and histopathological responses of a threatened freshwater fish, spotted murrel, Channa punctatus.

Two experiments, one short-term and one long-term, were conducted to elucidate the acute and chronic effects, respectively, of endosulfan exposure on the haemato-immunological and histopathological responses of Channa puncatatus. In the short-term study, fish were exposed to sublethal endosulfan (8.1 µg l(-1)) for 12, 24, 36, 48, 72 and 96 h. In the long-term study, fish were fed with normal diet and simultaneously either exposed to endosulfan (1.2 µg l(-1)) for 90 days or not. Results showed that the ascorbic acid levels in both the liver and the muscle decreased significantly (P < 0.05) by acute and chronic endosulfan exposure. The haemoglobin (Hb) level reduced significantly (P < 0.05) by 15.5% within 12 h of acute endosulfan exposure, further decreased by 25.8% after 24 h of exposure, however, thereafter the values increased and at the end of 72 h returned to normal levels. Almost similar trend was observed for the erythrocyte (RBC) count. The WBC count and the nitroblue tetrazolium (NBT) value showed a general increasing trend with increase in the duration of acute endosulfan exposure. The chronic exposure of C. punctatus to endosulfan significantly (P < 0.05) lowered the Hb level, RBC and WBC counts, NBT reduction value and the plasma parameters such as plasma protein, albumin (A) and globulin (G) compared with that of the control (except for A/G ratio). Endosulfan exposure also severely altered the liver histological structure. Overall results indicated that both short-term acute and long-term chronic endosulfan exposure had a significant impact on the haemato-immunological parameters and tissue histopathology of C. punctatus.

Yeast extract, brewer's yeast and spirulina in diets for Labeo rohita fingerlings affect haemato-immunological responses and survival following Aeromonas hydrophila challenge.

A feeding trial was conducted for 60 days to study the immunomodulatory role of three different immunostimulants yeast extract (YE), brewer's yeast (BY) and spirulina (SP) in Labeo rohita fingerlings. Four hundred and fifty fingerlings (avg. wt 3.35±0.15 g) were randomly distributed in ten treatments and fed with either of ten iso-nitrogenous and iso-caloric semi-purified diets, prepared with three incremental levels (1%, 2% and 4%) of different immunostimulants except the control. Growth parameters did not vary significantly (p>0.05) among the experimental groups. Haematology and serum parameters was performed before Aeromonas hydrophila challenge whereas respiratory burst activity was analysed following challenge. The respiratory burst activity, total leucocyte count, serum total protein and globulin was significantly higher (p<0.05) in YE 1% supplemented group. The survival (%) after challenging with A. hydrophila was also highest in the YE fed groups. The results indicate that among the different sources and levels of immunostimulants, YE at lower inclusion level is more effective in promoting the immune status of L. rohita fingerlings.

Biochemical and histological changes in the brain tissue of spotted murrel, Channa punctatus (Bloch), exposed to endosulfan.

The present experiment was conducted to establish the relationship between selected physiological parameters and histological responses of Channa punctatus brain tissue to endosulfan exposure. The fish (35.6 +/- 0.7 g) was exposed to sublethal endosulfan concentration (8.1 microg l(-1)) for a period of 12, 24, 36, 48, 72, and 96 h. Results showed that brain glucose level increased significantly after exposure, indicating a hyperglycemic state of the fish. Brain vitamin C level decreased with an increase in the exposure time. Acetylcholine esterase and adenosine triphosphatase enzyme activities also showed a significant reduction upon endosulfan exposure. Brain histopathology after 96 h endosulfan exposure showed that the apical lobe of the cerebrum (the only portion examined) had mild necrosis. Focal area of gliosis could be seen in the cerebrum, which were absent in the control fish. The results indicate that exposure of sublethal concentration of endosulfan to C. punctatus may have a direct effect on the histology of the fish's brain tissue, thereby affecting its metabolism.

DNA fingerprinting of Flavobacterium columnare using RAPD-PCR.

In the present study, DNA fingerprinting of eight strains of Flavobacterium columnare was done by random amplification of polymorphic DNA (RAPD) fingerprinting method. The strains were collected from Fish Health Management Division, Central Institute of Freshwater Aquaculture, Bhubaneswar, India. A total number of 160 primers were screened for RAPD-PCR, of which 10 primers yielded amplification with all the strains. The molecular weight of amplified bands varied from 0.29-2.63 Kb. The number of bands varied from 1 to 8. Unique band was seen with primer OPY-15 with molecular weight 0.75 Kb that can be used for epidemiological study. Genetic variability was investigated using NTSYS software. Highest genetic similarity was found between MS1 and MS3 followed by MS5 and MS7. Minimum genetic similarity was found between MS2 and MS8. Phylogenetic tree was constructed using UPGMA and neighbor joining methods.

Plasma levels of lipid peroxides in patients with Parkinson's disease.

Oxidative stress is implicated as a major factor for nigral neuronal cell death. Metabolic failure in antioxidant mechanisms could hypothetically facilitate the chemical processes that lead to lipid peroxidation. To elucidate whether elevated lipid peroxidation rates might increase risk of developing Parkinson's disease (PD), the Authors determined plasma levels of malondialdehyde (MDA) in 80 PD patients and 80 controls. There was a significant difference between the plasma MDA levels of PD patients and controls (7.48 +/- 1.55 vs 5.1 +/- 1.26 nmol/ml). Plasma MDA levels were inversely related to the age of the PD patients (r = -0.46; p < 0.01) and age of onset but in the control group, no such correlation was observed between the plasma MDA and age. However, there was no significant correlation between plasma MDA levels and the duration of disease, Hoehn and Yahr stages and the Unified Parkinson's Disease Rating Scale (UPDRS). Thus, the results suggest that high plasma lipid peroxidation rates might contribute as a risk factor for PD in West Bengal.

Characterization of IgM of Indian major carps and their cross-reactivity with anti-fish IgM antibodies.

Indian major carps (IMC), rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) were immunized with bovine serum albumin and the serum immunoglobulin M (IgM) was purified by affinity chromatography. The heavy and light chain of IgM of all the three species of IMC were about 88 and 26kDa, respectively. Anti-fish IgM antibody against all the three species were raised in mice and the reaction of anti-fish IgM antibodies with IgM of all the three species of IMC were studied by Western blot. The anti-fish IgM antibodies reacted strongly with the heavy chain of the same species against which it was raised while the reactions with the heavy chain of other species were milder indicating some degree of epitope sharing among the heavy chains of IgM of IMCs. However, there was no cross-reaction with the light chain of any of the IgM.

Microbial levan in the diet of Labeo rohita Hamilton juveniles: effect on non-specific immunity and histopathological changes after challenge with Aeromonas hydrophila.

A 60-day feeding trial was conducted to study the immuno-protective effect of microbial levan on Labeo rohita juveniles challenged with Aeromonas hydrophila. Six purified diets were prepared with different levels of microbial levan: control (no levan), T1 (Basal + 0.25%), T2 (Basal + 0.50%), T3 (Basal + 0.75%), T4 (Basal + 1%) and T5 (Basal + 1.25%), fed to six groups of fish in triplicate. Among the treatment groups the haemoglobin content and total leucocyte count were increased with a dietary supplementation of levan at 1% or more. An increasing trend for total erythrocyte count was observed with increasing level of dietary levan. Lower levan-supplemented groups showed a higher albumin/globulin ratio. As the levan supplementation was increased, there was a gradual increase in serum lysozyme activity and respiratory burst activity [nitroblue tetrazolium (NBT) assay] reduction values. The highest lysozyme activity and NBT were observed in the T5 group although this was similar to the T4 group (P > 0.05). No significant histo-architectural changes were associated with dietary levan levels. After challenge with A. hydrophila, moderately degenerated hepatocytes, oedema and leucocytic infiltration in parenchymatous tissues, and extensive haemorrhage and haemosiderosis in the kidney were observed in the control group. However, the T5 group supplemented with 1.25% levan showed infiltrating leucocytes in the liver while the kidney showed only moderate degeneration of renal tubules. The relative survival per cent of juveniles after challenge with A. hydrophila was the highest in the T5 group followed by T4. This suggests that microbial levan at 1.25% can be used as dietary immunostimulant for L. rohita juveniles.

Enhanced innate immune parameters in Labeo rohita (Ham.) following oral administration of Bacillus subtilis.

The present study assessed the use of Bacillus subtilis in fish as a probiotic. The bacterium was administered orally at three different doses 0.5 x 10(7) (T(2)), 1 x 10(7) (T(3)), 1.5 x 10(7) (T(4)) cfu/g feed to Labeo rohita for two weeks. The positive control group (T(1)) and negative control group (T(5,)) were fed feed without B. subtilis for the same period. On the 15th day blood and serum were sampled to determine respiratory burst activity (NBT assay), differential leukocyte counts (DLC) and serum bactericidal activity. Fishes were challenged intraperitoneally with Aeromonas hydrophila O:18 after two weeks in the treatment groups (T(2), T(3) and T(4)) and also in the positive control group(T(1)), while the negative control group (T(5)) was challenged with phosphate buffered saline (PBS, pH 7.2) only. The respiratory burst activity and DLC were assessed on the 3rd day post-challenge. B. subtilis treated fish showed significantly higher (P<0.05) respiratory burst activity and bactericidal activity during the pre-challenge compared with the control groups. The highest respiratory burst activity (0.37+/-0.03) and serum bactericidal activity were recorded in the group (T(4)) fed feed containing B. subtilis at 1.5 x 10(7)cfu/g feed. Granulocyte numbers were significantly higher (P<0.05) in treatment groups in comparison to the control in both the pre- and post-challenge periods. The result suggests that B. subtilis can enhance certain innate immune responses in rohu.

Haemato-immunological responses to dietary yeast RNA, omega-3 fatty acid and beta-carotene in Catla catla juveniles.

A preliminary study with 60 days feeding was conducted to study the immunomodulatory role of different immunostimulants like beta-carotene, omega-3 fatty acid and yeast-RNA in Catla catla fingerlings. Two hundred and sixty four fingerlings were randomly distributed into eight treatment groups with each of three replicates. Eight isonitrogenous (crude protein 34.12-35.40%) and isocaloric (458.41-461.48 kcal/100g) purified diets were prepared with graded level of beta-carotene, omega-3 fatty acid and yeast-RNA viz., Control (basal diet), T1 (Basal + 1% omega-3 fatty acid), T2 (Basal + 3% omega-3 fatty acid), T3 (Basal + beta-carotene), T4 (T1 + beta-carotene), T5 (T2 + beta-carotene), T6 (Basal + 0.4% yeast-RNA) and T7 (Basal + 0.8% yeast-RNA). The immunomodulatory effects of dietary immunostimulants were studied in terms of respiratory burst activity (NBT) of blood phagocytes, total leukocyte count, serum total protein, serum globulin, A/G ratio (A/G) and serum lysozyme activity. The respiratory burst activity of T7 group was significantly higher (p<0.05) than the other groups. Haemoglobin content, total erythrocyte count and serum albumin content did not vary among the treatment groups, whereas total leukocyte count, serum globulin content and serum lysozyme activity were found to be highest in T7 group. Relative survival percent after challenge with Aeromonas hydrophila was also highest in T7 (88.88%) group followed by T6 (75.06%) and T4 (66.66%) and the lowest in T2 group. It was observed that total leucocyte count, NBT and lysozyme activity of T2 group fed with high omega-3 fatty acid (3%) was less than (p<0.05) its lower counterparts T1 (1%) and control group. Based on the results of the present study, it concludes that supplementation of yeast-RNA at 0.8% registered higher immunological responses in C. catla juveniles. It is also observed that higher supplementation of omega-3 fatty acid (3%) in the diet causes immunosuppression in C. catla juveniles.

Effect of dietary supplementation of probiotic and vitamin C on the immune response of Indian major carp, Labeo rohita (Ham.).

The immunostimulatory effect of probiotics and vitamin C has been established in many systems including fish. An investigation was carried out to study the effect of dietary supplementation of a probiotic bacterium "Bacillus subtilis", vitamin C in the form of ascorbyl polyphosphate and their combination on the immune response of Indian major carp, rohu, (Labeo rohita Ham.) fingerlings fed for a period of 60 days. The total serum protein and globulin content was significantly higher (p<0.05) in probiotic (B. subtilis @ 10(8) CFU/g of the feed) fed group while the respiratory burst activity of blood neutrophils was significantly high in vitamin C (ascorbyl polyphosphate @100 mg per kg diet) fed group. The antibody level was significantly high in Bacillus subtilis treated group followed by the probiotic (B. subtilis @ 10(8) CFU/g of the feed) and ascorbyl polyphosphate (ascorbyl polyphosphate @100 mg per kg diet) combined group. The least percentage of mortality was recorded in B. subtilis treated group (25%) followed by 35 and 40% in ascorbyl polyphosphate treated and B. subtilis and ascorbyl polyphosphate combined groups, respectively.

Experimental vertical transmission of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) from brooders to progeny in Macrobrachium rosenbergii and Artemia.

White tail disease (WTD) is a serious problem in hatcheries and nursery ponds of Macrobrachium rosenbergii in India. Experiments were carried out to determine the possibility of vertical transmission of M. rosenbergii nodavirus (MrNV) and extra small virus (XSV) in M. rosenbergii and Artemia. Prawn broodstock inoculated with MrNV and XSV by oral or immersion challenge survived without any clinical signs of WTD. The brooders spawned 5-7 days after inoculation and the eggs hatched. The survival rate of larvae gradually decreased, and 100% mortality was observed at the post-larvae (PL) stage. Whitish muscle, the typical sign of WTD, was seen in advanced larval developmental stages. The ovarian tissue and fertilized eggs were found to be positive for MrNV/XSV by reverse transcriptase-polymerase chain reaction (RT-PCR) whereas the larval stages showed positive by RT nested PCR (nRT-PCR). In Artemia, reproductive cysts and nauplii derived from challenged brooders were normal and survival rates were within the expected range for normal rearing conditions. The reproductive cysts were found to be positive for MrNV/XSV by RT-PCR whereas the nauplii showed MrNV/XSV-positive by nRT-PCR. The PL of M. rosenbergii fed nauplii derived from challenged Artemia brooders died at 9 days post-inoculum with clinical signs of WTD.

Dietary microbial levan enhances cellular non-specific immunity and survival of common carp (Cyprinus carpio) juveniles.

A preliminary study with a 75days feeding trial was conducted to study the immunomodulatory effect of microbial levan on Cyprinus carpio juveniles. Five purified isonitrogenous and isocaloric diets with graded levels of levan, namely (T(1)) 0.1% levan, (T(2)) 0.2% levan, (T(3)) 0.5% levan, (T(4)) 1.0% levan, and a control group without levan were fed to five groups of fishes in triplicate. The total erythrocyte count and haemoglobin content was significantly (p<0.05) enhanced in the T(3) group, though the total leucocyte counts, total protein and A/G ratio did not show any significant difference (p>0.05) when compared with the control group. The respiratory burst activity (NBT) of blood phagocytes and lysozyme activity was also highest in T(3) group. The relative survival percentage after challenge with Aeromonas hydrophila was highest (100%) in the T(3) group followed by 83.33% and 66.67% in the T(2) and T(4) groups, respectively. This suggests that microbial levan at 0.5% can be used as a dietary immunostimulant for C. carpio juveniles. This is the first report on microbial levan having an immunomodulatory effect on C. carpio.

Correlation of lipid profile and carotid artery plaque as detected by Doppler ultrasound in ischaemic stroke patients--A hospital-based study.

Stroke is one of the leading causes of morbidity and mortality all over the world. Carotid plaque formation and intima media thickness can be a predictor of ischaemic stroke. In this regard studies from our country, are few and far between. This is a small hospital-based study to look in to this matter. We have assessed the intima media thickness of the common carotid as well as the internal and external carotid arteries by the ultrasound method. The lipid profiles were estimated and correlated with the intima media thickness. Results indicate that in the common as well as in the internal and external carotid arteries, the intima media thickness is a good predictor of ischaemic stroke. This thickness is also well correlated with the lipid levels in blood. Hence this non-invasive method can be used successfully to identify the high risk patients, prone to develop stroke.

Pre- and post-challenge immuno-haematological changes in Labeo rohita juveniles fed gelatinised or non-gelatinised carbohydrate with n-3 PUFA.

The combined effect of dietary carbohydrate type and n-3 PUFA (EPA+DHA) on pre- and post-challenge haemato-immunological responses in Labeo rohita juveniles was studied. Fish were fed for 67days with six different test diets containing either gelatinised (G) or non-gelatinised (NG) corn (43%) with three levels of n-3 PUFA (0.5%, 1.0% and 2.0%). During the pre-challenge period, significantly higher (P<0.05) NBT, serum lysozyme activity, total protein and globulin content was recorded in the NG carbohydrate fed groups. Highest NBT value was recorded in the groups fed with 1.0% n-3 PUFA, whereas the highest serum lysozyme activity (P<0.05) was recorded at either 0.5% or 2.0% n-3 PUFA fed groups in both the pre- and post-challenge period. Feeding of NG corn significantly increased the total leucocyte count, lysozyme activity, A/G ratio and decreased the total erythrocyte count, haemoglobin, serum total protein and globulin content of L. rohita juveniles during the post-challenge period. Similarly, feeding of n-3 PUFA at any level significantly increased the immunological parameters like lysozyme activity or A/G ratio, whereas total leukocyte count increased due to feeding of either 0.5% or 1.0% n-3 PUFA. The NBT and albumin values remained similar in both the pre- and post-challenge period. After challenge with Aeromonas hydrophila, the highest survival was recorded in the NG carbohydrate fed groups, whereas the lowest survival was recorded in the highest level of n-3 PUFA fed group irrespective of dietary carbohydrate type. Thus, a high level of G carbohydrate as well as n-3 PUFA is found to be immunosuppressive in L. rohita juveniles. NG carbohydrate supplemented with 1.0% n-3 PUFA is found to be optimum to enhance the immunity in L. rohita juveniles.

The immunomodulatory effects of tuftsin on the non-specific immune system of Indian Major carp, Labeo rohita.

The purpose of this study was to determine if injections of different dosages of tuftsin would enhance the immune response and disease resistance against the infections due to the opportunistic pathogens Aeromonas hydrophila and Edwardsiella tarda in Labeo rohita fingerlings. Hence, four different dosages of tuftsin in PBS suspension at the rate of 0, 5, 10, 15 mg kg(-1) body weight of fish were injected intraperitoneally to the fingerlings of L. rohita at 2-week intervals for four times. After every 2-week interval, different serum biochemical, haematological and immunological parameters of fish were evaluated. Biochemical and haematological parameters including serum total protein content, albumin content, globulin content, albulin:globulin ratio, glucose content, leucocyte counts etc.; cellular immune parameters including superoxide anion production, phagocytic activities, lymphokine production index etc.; humoral immune parameters including lysozyme activity, complement activity, serum bactericidal activity etc., in the fish were evaluated after every 2-week interval. After 56 days, fish were divided into two subgroups under each major treatment group for challenge with two pathogens A. hydrophila and E. tarda. The mortality (%) and agglutinating antibody titre was recorded on 28th day post challenge. Most of the immune parameters including leucocyte count, phagocytic ratio, phagocytic index, lysozyme activity, complement activity, and serum bactericidal activity were significantly (p

Dietary high protein and vitamin C mitigate stress due to chelate claw ablation in Macrobrachium rosenbergii males.

Stress due to claw ablation was tested in Macrobrachium rosenbergii males. Dietary high protein and vitamin C were supplemented for amelioration of stress. We used four different treatments: fed with 25% protein and a normal dose (0.12%) of vitamin C (T(1)); 35% protein and a normal dose (0.12%) of vitamin C (T(2)); 25% protein and a high dose (0.24%) of vitamin C (T(3)); and high protein 35% and a high dose (0.24%) of vitamin C (T(4)) for 30 days. All test prawns (T(1) to T(4)) were subjected to ablation of their second chelate legs after the 15th day of the feeding trial. A control treatment was maintained without claw ablation and fed with 25% protein. Haemolymph glucose, hepatopancreatic glycogen, muscle ascorbate and enzyme activities (glucose 6 phosphatase (G6Pase), fructose-1,6-bisphosphatase (FBPase), lactate dehydrogenase (LDH), Alanine aminotransferase (ALT) in hepatopancreas) were tested at different recovery periods (0, 6, 24 h, 7 and 14 days). Results indicate a high glucose level immediately after claw ablation and a concomitant increase in gluconeogenic enzymes (G6Pase and FBPase). However, glycogen reserves were regained in the treatments due to claw ablation stress after 24 h. LDH and ALT activity decreased in the hepatopancreas of M. rosenbergii up to 24 h after claw ablation. Overall results indicate that claw ablation is stressful to M. rosenbergii and high protein and vitamin C diet may mitigate stress due to claw ablation.

Effect of dietary carbohydrate on haematology, respiratory burst activity and histological changes in L. rohita juveniles.

A feeding trial was conducted for 60 days to delineate the effect of dietary carbohydrate on the haematology, respiratory burst activity and histology in Labeo rohita juveniles. One hundred and forty four fish (av. wt. 10+/-0.15 g) were randomly distributed into 12 treatment groups with each of two replicates. Twelve semi-purified diets with either 35% or 28% of crude protein were prepared with different amylase concentrations (0, 50, 100 and 150 mgkg(-1)) and starch type (gelatinised, G/non-gelatinised, NG) designated as T1 (NG, 35% CP, 0 mgkg(-1) amylase), T2 (G, 35% CP, 0 mgkg(-1) amylase), T3 (NG, 28% CP, 50 mgkg(-1) amylase), T4 (NG, 35% CP, 50 mgkg(-1) amylase), T5 (G, 28% CP, 50 mgkg(-1) amylase), T6 (G, 35% CP, 50 mgkg(-1) amylase), T7 (NG, 28% CP, 100 mgkg(-1) amylase), T8 (NG, 35% CP, 100 mgkg(-1) amylase), T9 (G, 28% CP, 100 mgkg(-1) amylase), T10 (G, 35% CP, 100 mgkg(-1) amylase), T11 (NG, 28% CP, 150 mgkg(-1) amylase) and T12 (NG, 35% CP, 150 mgkg(-1) amylase). The fish were acclimatised to the experimental conditions for 15 days during which time the control diet was fed. The blood haemoglobin percentage and RBC count was not significantly different (P>0.05) among the various treatments. Highest WBC count, total plasma protein, serum globulin and respiratory burst activity was found at 50 mgkg(-1) dietary amylase supplementation, whereas no variation (P>0.05) was found at 0, 100 and 150 mgkg(-1) dietary amylase supplementation. There was no difference (P>0.05) in the serum albumin and AG ratio among the experimental groups. Feeding either gelatinised or non-gelatinised carbohydrate did not affect the histological structure of the liver, kidney and intestine except in the T4 and T7 groups. The T4 group showed hyperplasia of the intestine and moderate vacuolation in hepatic cells, whereas the T7 group showed hyperplasia of the intestine. Non-gelatinised carbohydrates (46%) along with supplementation with 50 mgkg(-1) amylase stimulated the immune system in L. rohita juveniles. But supplementation of amylase to the gelatinised carbohydrate had no immunostimulating effect.