On valorization of solvent extracts of Terminalia arjuna (arjuna) upon DNA scission and free radical scavenging improves coupling responses and cognitive functions under in vitro conditions.

Chronic diseases have been treated using the phytochemical concepts of ethnomedicinal plant-derived herbal products. Terminalia arjuna, a significant ethnomedicinal plant, was revisited and reconnoitred for antioxidant, free radical scavenging, and DNA nicking inhibiting activity under H2O2 conditions using 21 solvent extracts. Ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid, and nitrous oxide scavenging (%) were found to have a strong positive association and interaction (PCA 1 explains 84.54% variation) with ethanol bark (Etoh-AB) (Meoh-AF). TPC (144.67-1794 µg/mL GAE) and TFC (2.5-34 µM Fe(II)/g were highest in Etoh-AB. In a pattern of combined solvent extracts, Etoh-AB had the highest antioxidant capacity, accompanied by Etoh-AL ≥ Meoh-AB ≥ Dw-AF. With R2 = 0.94, the DNA nicking inhibition behaviour parameters relative front, relative quantity, band (%), and lane (%) formed a positive significant (p < 0.01) connection. For the first time, we show that Etoh-AB nicks supercoiled, circular plasmid DNA in a way that is comparable to normal antioxidants. Normal antioxidants with the ability to prevent DNA nicking include Butylated hydroxy anisole < Butylated hydroxy toluene < ascorbic acid < and Gallic acid. Gallic acid (m/z 170.0208 g/mol) and Ellagic acid (m/z 302.0063 g/mol were present in high concentrations in solvent extracts. 0.48 mg was found to be the effective concentration for inhibiting relative DNA nicking. The current study is the first of its kind to show that steroid concentrations are higher in bark fractions of acetone, ethanol, and methanol. Furthermore, T. arjuna solvent extracts provide a wealth of information on phytochemical profiling, antioxidant ability, and DNA nicking inhibition, which may be useful for exploring the natural way and further research to develop a remedy against geriatric chronic disease. Despite the fact that ethanol is very close to methanol in terms of solvent toxicity, the current study identified it as the preferred solvent. Thus, the current research revisits previous studies and explores the potentiality of non-polar and polar aprotic and polar protic solvent systems, which lend credence to bioactive compounds that may be useful in isolating and formulating safe and cost effective herbal medicament for livestocks and aquaculture, and drugs for deoxygenerative human diseases, and can also be investigated further to instil environmental frugality.

Physio-metabolic and haematological changes of Labeo rohita fed with graded level of de-oiled rice bran-based diet.

A 60-day feeding trial was conducted with six experimental diets containing 260 g/kg crude protein and 70 g/kg lipid with varying level of de-oiled rice bran (DORB), viz., 33 (T33), 38 (T38), 43 (T43), 48 (T48), 53 (T53) and 58% (T58) to assess the digestive, metabolic and haematological responses in Labeo rohita. One hundred and eighty (180) fingerlings with ten fish per tank having an average weight of 9.8 ± 0.5 g were randomly distributed within six treatments in triplicates following a completely randomised design. The amylase and lipase activities in the whole intestine did not vary significantly (P > 0.05), but protease activity varied significantly with respect to DORB level in the diet. At higher (53 and 58%) inclusion level of DORB, the protease activities were lower. T33, T38 and T43 groups showed significantly (P < 0.05) higher aspartate aminotransferase (AST) activity compared to the other groups, whereas lowest activity was recorded in the group fed with highest inclusion level (58%). The oxidative stress enzyme like superoxide dismutase (SOD), catalase and glutathione-S-transferase exhibited significantly higher activities in liver of T58 group of L. rohita. However, the activity of catalase and SOD showed no significant variation in gill tissue. The haematological parameter values such as red blood cell (RBC), white blood cell (WBC), haemoglobin (Hb) and packed cell volume (PCV) were lower in the highest DORB fed group (T58) as compared to their lower DORB inclusion counterparts. No change in NBT value was recorded irrespective of DORB inclusion level in the diet. Hence, from these results, it can be concluded that the dietary inclusion of DORB up to 43% in the diet of L. rohita at 26% crude protein and 7% lipid level is best for the optimal activity of digestive, metabolic and haematological parameters.

Dietary L-Tryptophan potentiates non-specific immunity in Labeo rohita fingerlings reared under elevated temperature.

At present environmental scenario global climate change is a reality and its affect all living organism including fish. The aquatic ecosystem is the most affected system as it is the biggest sink for global warming and elevated temperature and obviously affects all the aquatic life forms. With this hypothesis an experiment was conducted to evaluate the effect of elevated temperature on Labeo rohita fingerlings and potential of dietary L-tryptophan (TRP) in mitigating the effects elevated temperature and enhancing the non-specific immunity. Seven hundred and twenty fishes were randomly distributed in three different thermal groups each with three replicates. The thermal groups were ambient temperature (26 °C), 34 and 38 °C. Then each thermal groups were fed with four different formulated diets containing 0. 0.36%, 0.72% and 1.44% TRP. The effect of dietary TRP supplementation was studied on stress responses, such as cortisol, blood glucose, histopathological changes in liver and kidney and immuno-hematological changes such as red blood cell count (RBC), haemoglobin (Hb), white blood cell count (WBC), lysozyme, nitroblue tetrazolium (NBT), total serum protein, albumin, globulin and albumin-globulin ratio. Subsequently the treated fish were subjected to challenge test with Aeromonas hydrophila. In the present study, primary stress markers were noticeably (p < 0.01) elevated with temperature stress and levels were reduced with nutritional supplementation of TRP. Similarly, immuno-hematological parameters were altered with the exposure of temeparture stress and got improved with dietary TRP supplementation. Results of the present study suggest that dietary supplementation of 1.44% tryptophan has definitive role in the mitigation of temperature stress and gives protection against bacterial infection to L. rohita.

Effect of dietary supplementation of l-tryptophan on thermal tolerance and oxygen consumption rate in Cirrhinus mrigala fingerlings under varied stocking density.

A 60 day feeding trial was conducted to study the effect of dietary l-tryptophan on thermal tolerance and oxygen consumption rate of freshwater fish, mrigala, Cirrhinus mrigala reared under ambient temperature at low and high stocking density. Four hundred eighty fingerlings were distributed into eight experimental groups. Four groups each of low density group (10 fishes/75L water) and higher density group (30 fishes/75L water) were fed a diet containing 0, 0.68, 1.36 or 2.72% l-tryptophan in the diet, thus forming eight experimental groups namely, Low density control (LC) (basal feed +0% l-tryptophan); LT1 (basal feed+0.68% l-tryptophan); LT2 (basal feed+1.36% l-tryptophan); LT3 (basal feed+2.72% l-tryptophan); high density control (HC) (basal feed+0% l-tryptophan); HT1 (basal feed+0.68% l-tryptophan); HT2 (basal feed+1.36% l-tryptophan); and HT3 (basal feed+2.72% l-tryptophan) were fed at 3% of the body weight. The test diets having crude protein 34.33±0.23 to 35.81±0.18% and lipid 423.49±1.76 to 425.85±0.31KCal/100g were prepared using purified ingredients. The possible role of dietary l-tryptophan on thermal tolerance and oxygen consumption rate was assessed in terms of critical thermal maxima (CTMax), critical thermal minima (CTMin), lethal thermal maxima (LTMax) and lethal thermal minima (LTMin). The CTMax, CTMin, LTMax and LTMin values were found to be significantly higher (p<0.05) in the treatment groups with CTMax 42.94±0.037 (LT2); LT Max 43.18±0.070 (LT2); CTMin 10.47±0.088 (LT2) and LTMin 9.42±0.062 (LT3), whereas the control group showed a lower tolerance level. The same trend was observed in the high density group (CTMax 42.09±0.066 (LT3); LTMax 43 23±0.067 (HT3); CTMin 10.98±0.040 (HT3) and LTMin 9.74±0.037 (HT3). However, gradual supplementation of dietary l-tryptophan in the diet significantly reduced the oxygen consumption rate in both the low density group (Y=-26.74x+222.4, r²=0.915) and the high density group (Y=-32.96x+296.5, r²=0.8923). Dietary supplementation of l-tryptophan at a level of 1.36% improved the thermal tolerance level and reduced the oxygen consumption rate in C. mrigala fingerlings.

Stress mitigating and growth enhancing effect of dietary tryptophan in rohu (Labeo rohita, Hamilton, 1822) fingerlings.

An experiment was conducted to study the stress mitigation and growth enhancing role of dietary L-tryptophan (TRP) under thermal stress in rohu, Labeo rohita fingerlings for 45 days. Seven hundred and twenty fishes were distributed in three major groups that are ambient temperature (26 °C), 34 and 38 °C in triplicate following a complete randomized design. Acclimation of fishes to 34 and 38 °C over average ambient temperatures were carried out at 1 °C/day. Each group was fed with a diet supplemented with 0, 0.36, 0.72 or 1.42 % L-TRP. Results showed that blood glucose and serum cortisol level were found to be significantly higher (p < 0.05) in the higher temperature groups than the ambient temperature group. Similarly, aminotransferase, lactate dehydrogenase, malate dehydrogenase, CAT, superoxide dismutase activities were found to be significantly higher (p < 0.05) in the control groups (0 % L-TRP) and decreasing activities of these enzymes were observed with the increasing level of dietary L-TRP. In different temperature groups, L-TRP-supplemented groups were found to have higher (p < 0.05) growth, RGR and PER. The results obtained in the present study indicate that dietary L-TRP mitigates thermal stress and enhances growth. From the present study, we can conclude that dietary supplementation of L-TRP at the 0.72 % level in the diet is found to be optimum to reduce thermal stress even up to 38 °C in rohu, L. rohita. The baseline data obtained here could be useful for the farmers to formulate feeds to culture the fish in different agro-climatic zones.

Supplementation of microbial levan in the diet of Cyprinus carpio fry (Linnaeus, 1758) exposed to sublethal toxicity of fipronil: effect on growth and metabolic responses.

A 60-day feeding trial was conducted to study the effect of dietary microbial levan on growth performance and metabolic responses of Cyprinus carpio fry exposed to sublethal dose (1/10th LC50) of fipronil [(±)-5-amino-1-(2,6-dichloro-α,α,α-trifluoro-p-tolyl)-4-trifluoromethylsulfinylpyrazole-3-carbonitrile]. Two hundred and twenty five fry were randomly distributed in five treatments in triplicates. Four purified diets were prepared with graded levels of microbial levan. Five different treatment groups were levan control L0P0 (basal feed + 0 % levan without exposure to pesticide); pesticide control L0P1 (basal feed + 0 % levan with exposure to pesticide); L0.25P1 (basal feed + 0.25 % levan with exposure to pesticide); L0.50P1 (basal feed + 0.50 % levan with exposure to pesticide); and L0.75P1 (basal feed + 0.75 % levan with exposure to pesticide). Weight gain% and specific growth rate were significantly higher (p < 0.05) in levan fed groups compared to their non-levan fed counterpart. Highest (p < 0.05) content of ascorbic acid in muscle, liver and brain tissues was observed with higher level of dietary levan. Glucose-6-phosphate dehydrogenase activity decreased with the increasing level of dietary levan in the liver and muscle. Aspartate aminotransferase activity exhibited a second order polynomial relationship with the dietary levan, both in liver (Y = -1.001x² + 5.366x + 5.812, r² = 0.887) and muscle (Y = -0.566x² + 2.833x + 6.506, r² = 0.858) while alanine aminotransferase activity showed third order polynomial relationship both in liver (Y = 1.195x³ - 12.30x² + 35.23x + 9.874, r² = 0.879) and muscle (Y = 0.527x³ - 8.429x² + 31.80x + 8.718, r² = 0.990). Highest (p < 0.05) superoxide dismutase activity in gill was observed in the group fed with 0.75 % levan supplemented diet. Overall results indicated that dietary microbial levan at 0.75 % in C. carpio fry ameliorated the negative effects of fipronil and augmented the growth.

Nitrite-induced alterations in sex steroids and thyroid hormones of Labeo rohita juveniles: effects of dietary vitamin E and L-tryptophan.

An experiment was conducted to study the effect of sub-lethal nitrite exposure on sex steroids (testosterone and estradiol), cortisol and thyroid hormones (T3 and T4) of Labeo rohita juveniles. Fishes previously fed with normal or elevated levels of vitamin E (VE) and tryptophan for 60 days were exposed to sub-lethal nitrite for another 45 days with same feeding regime. There were nine treatment groups, viz. VE0TRP0-N, VE0TRP0+N, VE100TRP0-N, VE100TRP0+N, VE100TRP0.75+N, VE100TRP1.5+N, VE150TRP0+N, VE300TRP0+N and VE200TRP1+N. Except the groups VE0TRP0-N and VE100TRP0-N, all other groups were exposed to nitrite. At the end of the 45 days of nitrite exposure, serum samples were assayed for sex steroids, cortisol and thyroid hormones. The serum T3 and T4 levels decreased to the extent of 84.5 and 94.06%, respectively, upon nitrite exposure. Dietary supplementation with additional amounts of VE and tryptophan appears to reduce the decline of the production of T4. The serum testosterone and estradiol decreased 97.31 and 92.86%, respectively, upon nitrite exposure. Supplementation with additional amounts of VE was found to reverse nitrite-induced inhibition of testosterone and estradiol production. Serum cortisol increased upon nitrite exposure and unexposed (VE100-N) group showed lower levels, which were comparable to groups fed with elevated levels of VE. The overall results of the present study revealed that environmental nitrites have a negative impact on steroidogenesis, which can be overcome by dietary supplementation of elevated amounts of VE (minimum of 150 mg VE Kg diet(-1)) and to a lesser extent by tryptophan (only at the level of 1.5% of the diet).

Physiological responses of dietary tryptophan fed Labeo rohita to temperature and salinity stress.

Two experiments were conducted to elucidate the possible effects of dietary L-tryptophan (TRP) in Labeo rohita based on growth performance and physio-biochemical responses. In the experiment I, a 60-day feeding trial was carried out to elucidate the effects of dietary TRP enrichment on growth performance and physio-biochemical responses. In the experiment II, the TRP pre-fed L. rohita, from experiment I, was exposed to temperature and salinity stress to evaluate stress-mitigating efficacy of TRP. In L. rohita, dietary supplementation of TRP showed significant effect on weight gain percentage and feed conversion ratio but not on blood glucose. A significant increase in RNA content and RNA/DNA ratio upon TRP supplementation was observed and was positively correlated with growth performance. The results of experiment II indicated that weight gain percentage, serum T3 and T4 levels were significantly reduced in groups that were exposed to temperature and salinity stress and fed diets without TRP supplementation. However, dietary supplementation of TRP significantly augmented weight gain percentage in stress-exposed groups. Tryptophan supplementation helped in bringing back T3 and T4 levels comparable with control. A significant increase in superoxide dismutase, catalase, Adenosine triphosphatase, blood glucose and serum cortisol was observed in temperature- and salinity-exposed groups fed without TRP-supplemented diets. However, TRP supplementation was found to be effective in restoring the above parameters. The results of these experiments suggest that dietary TRP supplementation augments growth, lowers energy demand and helps in mitigating thermal and salinity stress in L. rohita.

Effects of dietary pyridoxine on haemato-immunological responses of Labeo rohita fingerlings reared at higher water temperature.

A sixty-day feeding trial was carried out to elucidate the effect of dietary pyridoxine (PN) on hemato-immunological parameters in Labeo rohita fingerlings exposed to an elevated temperature (ET) of 33 °C. Two hundred and seventy fingerlings (6.71 ± 0.32 g) were randomly distributed into six treatments in triplicates (15 fish/tank). Five iso-nitrogenous (356.3 ± 2.7 g/kg crude protein) purified diets were prepared with graded levels (0, 10, 50, 100 and 200 mg/kg diet) of PN. Six treatment groups were P10T26 (10 mg PN + ambient temperature (26 °C), P0T33 (0 mg PN + ET), P10T33 (10 mg PN + ET), P50T33 (50 mg PN + ET), P100T33 (100 mg PN + ET) and P200T33 (200 mg PN + ET). The responses of L. rohita fingerlings to dietary PN were assessed in terms of growth and by analysing some hemato-immunological parameters. When PN supplementation increased from 10 to 200 mg/kg diet, a significant improvement in weight gain percentage was observed. Erythrocyte (RBC) count, leucocyte (WBC) count, haemoglobin content and respiratory burst activity were significantly (p < 0.05) higher in 100 mg PN/kg diet-fed group than their non-PN-fed counterpart. Also, serum albumin, globulin and lysozyme activity were found significantly (p < 0.05) higher in PN-fed groups. The overall results indicated that dietary PN supplementation at 100 mg/kg diet may reverse the negative effects caused by ET and may protect the haemato-immunological status of L. rohita fingerlings reared at higher water temperature.

Water soaking and exogenous enzyme treatment of plant-based diets: effect on growth performance, whole-body composition, and digestive enzyme activities of rohu, Labeo rohita (Hamilton), fingerlings.

A 2 × 2 × 2 factorial experiment was conducted to delineate the main effect of water soaking of plant ingredients, phytase, cellulase, and their interactions on the growth and digestive enzyme activities of Labeo rohita fingerlings. Two basal diets were prepared using water-soaked (S) or unsoaked (US) plant-based ingredients. Feed of US ingredients was supplemented with phytase (U kg(-1)) and cellulase (%) at the level of 0, 0 (C(us)); 500, 0 (T(1)); 0, 0.2 (T(2)); 500, 0.2 (T(3)), and feed of S ingredients at 0, 0 (C(s)); 500, 0 (T(4)); 0, 0.2 (T(5)), and 500, 0.2 (T(6)), respectively. Three hundred and sixty fingerlings were randomly distributed into eight treatments, each with three replicates. Soaking of the ingredients for 24 h significantly reduced the tannin content. However, feeding of S diets did not improve the fish growth. Highest performance was recorded in the T(3) group. A significant interaction between dietary phytase and cellulase was observed for apparent net protein utilization. Tissue crude protein, ether extract, and ash content of the fingerlings were observed highest in the T(3) group. Activities of amylase, protease, and lipase were recorded highest in the T(3) group. Results suggested that soaking of plant-based ingredients reduces tannin content; however, growth and digestive enzyme activities of group fed soaked diet were not improved, possibly due to leaching of soluble nutrients. Probably, a shorter duration soaking may be effective in reducing tannin content and avoiding nutrients leaching.

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.

Haemato-immunology and histo-architectural changes in Labeo rohita fingerlings: effect of dietary aflatoxin and mould inhibitor.

A feeding trail of 60 days was conducted for delineating the effect of dietary aflatoxin (AFB1) with or without supplementation of a mixture of mould inhibitor (0.25% clove oil + 0.32% sodium propionate) on haematology, respiratory burst activity and histology of Labeo rohita fingerlings. Three hundred and sixty fishes (avg. wt. 1.48-1.54 g) were randomly distributed into eight treatment groups. Eight experimental diets with four different levels of aflatoxin (0, 10, 20 and 40 ppb) with or without mould inhibitor were prepared. Haematological parameters like total serum protein, albumin, globulin and A:G ratio were significantly (P < 0.05) reduced with increasing levels of aflatoxin in the diet. However, supplementation of mould inhibitor showed enhanced values when compared to non-supplemented counter parts suggesting ameliorating effects of mould inhibitor on aflatoxin. Total leucocyte count was higher in aflatoxin-treated groups. Histological observations were complementary to haematological parameters. Respiratory burst activity was significantly (P < 0.05) decreased in higher aflatoxin-treated groups but not affected significantly (P > 0.05) due to inclusion of inhibitor alone and/or interaction of aflatoxin level and inhibitor in the diet. From this study, it was concluded that up to 20 ppb aflatoxin level in the diet the haemato-immunological parameters are protected.

Dietary high protein and vitamin C mitigates endosulfan toxicity in the spotted murrel, Channa punctatus (Bloch, 1793).

The amelioration effect of dietary high protein and vitamin C against stress was evaluated in spotted murrel, Channa punctatus, exposed to endosulfan. Two hundred and forty fish (average weight: 27.01 g/fish), distributed equally into 4 different groups (control, T(1), T(2), and T(3)), each with 6 replicates were fed with control (40% crude protein, CP and 0.1% vitamin C), T(1) (40% CP and 0.1% vitamin C), T(2) (50% CP and 0.1% vitamin C), and T(3) (50% CP and 0.2% vitamin C) diets for 90 days. Groups fed T(1), T(2), and T(3) diets were exposed to sublethal endosulfan concentration, whereas the control was maintained without endosulfan exposure. Results indicated significant reduction in the growth performance, survival, and activities of lactate dehydrogenase (liver and muscle), malate dehydrogenase (liver and muscle), enzymes of protein metabolism (aspartate amino transferase in liver and alanine amino transferase in liver and muscle), acetyl choline esterase (brain), alkaline phosphatase activity (liver), and ATPase (gill) enzymes of group fed control diet and exposed to endosulfan. However, endosulfan exposed fish fed high CP and vitamin C diet exhibited significant (P<0.05) improvement in their growth performance and metabolic enzyme activities. Further, high CP and vitamin C diet reduced endosulfan accumulation in the muscle. Overall results indicate that vitamin C (0.2%) supplementation in high CP (50%) diet improves growth, metabolism, and reduce endosulfan bioaccumulation in C. punctatus.

Non-gelatinized starch influences the deposition of n-3 fatty acids in the muscle of a tropical freshwater fish, Labeo rohita.

A 60-day feeding trial was conducted to study the influence of gelatinized (G) to non-gelatinized (NG) starch ratio in the diet on fatty acids profiles and oxidative status in Labeo rohita fingerlings. Two hundred and thirty-four fingerlings (average weight: 2.53 g) were distributed in six treatment groups with each of three replicates. Six semi-purified diets either containing NG and/or G corn starch (42.43%) viz., T(1) (100% NG and 0% G starch), T(2) (80% NG and 20% G starch), T(3) (60% NG and 40% G starch), T(4) (40% NG and 60% G starch), T(5) (20% NG and 80% G starch) and T(6) (0% NG and 100% G starch) was fed to respective groups. Catalase, superoxide dismutase and malic enzyme activities decreased linearly with the increasing level of G starch, whereas reverse trend was found for glucose-6-phosphate dehydrogenase. Total saturated fatty acids in muscle increased with the increasing level of G starch in the diet. Total n-3 fatty acids decreased linearly with the increasing level of G starch in the diet. Among the n-3 fatty acids, linolenic acids content was more in NG starch fed group. Similarly, eicosapentaenoic acid contents gradually decreased with increasing level of G starch content. The n-6/n-3 ratio was higher in G starch fed group. This suggests that dietary starch type may be manipulated for quality improvement of fish flesh.

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.

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.

Trigonella foenum graecum (fenugreek) seed extract as an antineoplastic agent.

The antineoplastic effect of Trigonella foenum graecum seed extract has been evaluated in the Ehrlich ascites carcinoma (EAC) model in Balb-C mice. Intra-peritoneal administration of the alcohol extract of the seed both before and after inoculation of EAC cell in mice produced more than 70% inhibition of tumour cell growth with respect to the control. Treatment with the extract was found to enhance both the peritoneal exudate cell and macrophage cell counts. The extract also produced a significant antiinflammatory effect. We report here the antiinflammatory and antineoplastic effects, of Trigonella foenum graecum seed extract.

Triterpenoid saponins from Mimusops elengi.

A novel minor triterpenoid saponin mimusin (3-O-[beta-D-glucopyranosyl- (1-->6)-beta-D-glucopyranosyl]-2 beta,3 beta,6 beta,23-tetrahydroxyolean- 12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1-->3)-beta-D-xylopyranosyl-(1-->4)-alpha- L- rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside was isolated from the seeds of Mimusops elengi, in addition to two known triterpenoid saponins, Mi-saponin A and 16 alpha-hydroxy Mi-saponin A. The structure of the minor saponin was established by comparing its 13C NMR and LS-MS linked-scan, ESI-MS data with FAB-MS of the mimusopsin isolated earlier from the same source.

Triterpenoid saponins from Gymnema sylvestre.

Besides six known gymnemic acids, four new tritepenoid saponins, gymnemasins A, B, C and D, isolated from the leaves of Gymnema sylvestre, were identified as 3-O-[beta-D-glucopyranosyl(1-->3)-beta-D-glucuronopyranosyl]-22-O- tigloyl- gymnemanol, 3-O-[beta-D-glucopyranosyl (1-->3)-beta-D-glucuronopyranosyl]-gymnemanol, 3-O-beta-D-glucuronopyranosyl-22-O-tigloyl-gymnemanol and 3-O-beta-D-glucuronopyranosyl-gymnemanol, respectively. The aglycone, gymnemanol, which is a new compound, was characterized as 3 beta, 16 beta, 22 alpha, 23, 28-pentahydroxyolean-12-ene.

Anti-inflammatory triterpene saponins of Pithecellobium dulce: characterization of an echinocystic acid bisdesmoside.

A new bisdesmodic triterpenoid saponin, dulcin was isolated from the seeds of Pithecellobium dulce and was identified as 3-O-[beta-D-glucopyranosyl (1-->2)-alpha-L-arabinopyranosyl]- 28-O-[beta-D-xylopyranosyl (1-->6)-beta-D-glucopyranosyl]-echinocystic acid. The known oleanolic acid saponin PE, oleanolic acid 3-O-beta-D-glucopyranosyl (1-->2)-alpha-L-arabinopyranoside was also obtained. The structural features were elucidated by a combination of spectroscopic methods and some chemical transformations.