Browsing by Author "SINDHU, S"
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ThesisItem Open Access BIOACTIVITY OF CAROTENOIDS FROM SHRIMP SHELL WASTE(KUFOS, 2010-08-30) SINDHU, SABSTRACT Shrimp processing waste is the single largest industrial waste in the country causing diverse environmental problems. A study was carried out to assess the extractability of astaxanthin from shrimp waste in different organic solvents and vegetable oils. Extraction was tried using wet and dried waste, with and without deproteinisation. Waste was subjected to deproteinisation using alkali and enzyme (pancreatin). The different solvent systems tried were ether:acetone:water (15:75:10 v/v/v), acetone, hexane:isopropanol (3:2 v/v) and 90% acetone v/v. Astaxanthin in the extract was quantified by measuring the OD at 470 nm in hexane. Extraction was also done using vegetable oils viz. coconut oil, soybean oil and sunflower oil. Quantification of astaxanthin in pigmented oil was done by measuring the absorbance at 485 nm using 2155 as extinction coefficient. Astaxanthin yields from deproteinised samples were significantly lower than those from non deproteinised samples. The highest astaxanthin yield of 87.14 ± 4.55μg/g was obtained with non deproteinised wet waste extracted using acetone. The astaxanthin yield was significantly lower when oil was used as the extraction medium. Of the three oils coconut oil gave the highest yield. The results showed that acetone is the best solvent for extracting astaxanthin from shrimp shell waste in wet condition. The astaxanthin content in Aristeus alcocki shell waste is double that of Pandalus borealis shell waste, which is currently used as the commercial source of astaxanthin. The deep sea species Aristeus alcocki can thus be considered as a better source of astaxanthin for commercial exploitation than Pandalus borealis. TLC analysis of the shell waste extract showed that it contains free astaxanthin, astaxanthin monoester and astaxanthin diester in the ratio 1:1:2. GLC identification of the fatty acids esterified with astaxanthin revealed that saturated fatty acids, MUFA and PUFA are in the ratio 5:3:2 in monoester, whereas in diester they are in the ratio 4:3:3. The main fatty acids in monoester and diesters are palmitic acid, oleic acid, stearic acid and PUFAs: DHA and EPA. The in vitro antioxidant activity of the astaxanthin extract showed significant hydroxyl radical scavenging activity, superoxide anion scavenging activity and inhibition of lipid peroxidation. The IC50 values obtained were 56.43 ± 1.06 ng/ml, 27.91 ± 0.54 ng/ml and 26.54 ± 0.42 ng/ml, respectively. The antioxidant activity of astaxanthin from Aristeus alcocki was obtained at nanogram levels. This powerful antioxidant function may be due to the unique molecular structure of astaxanthin and synergistic effect of astaxanthin and PUFAs present in the astaxanthin monoester and diester fractions. The astaxanthin extract from shrimp shell waste significantly reduced carageenan induced paw edema in mice, percentage inhibition being 47.83 and 67.11 percent at astaxanthin concentrations of 0.5 mg/kg body weight and 1.0 mg/kg body weight, respectively. The inhibition of inflammation at 1.0mg/kg body weight was greater than that produced by the standard reference drug diclofenac. Cardioprotective effect of astaxanthin was examined in isoproterenol induced myocardial infarction in rats. Levels of diagnostic marker enzymes, LDH, CPK, GOT, GPT, CK, CK-MB in plasma, lipid peroxides, ascorbic acid, reduced glutathione and the activities of glutathione-dependent antioxidant enzymes GPx, GR, GST and antiperoxidate enzymes CAT, SOD and the membrane bound enzyme Na+ - K+ ATPase in the heart tissues of experimental groups of rats were determined. The prior administration of astaxanthin @ 10mg/kg feed for 45 days significantly prevented the isoproterenol-induced elevation in the levels of diagnostic marker enzymes in plasma, induction of lipid peroxidation and alterations in the level of reduced glutathione and in the activities of glutathione dependent antioxidant enzymes and antiperoxidative enzymes of experimental rats. Feeding astaxanthin caused a decrease in the inhibition of Na+ - K+ ATPase activity against isoproterenol induced myocardial infarction. The powerful cardioprotective effect of astaxanthin can be attributed to the multiple independent mechanisms viz. antioxidant effects, singlet oxygen quenching ability and inhibition of lipid peroxidation of membranes, increased functional gap junctional intercellular communication, anti-inflammatory effects etc. Immunostimulatory action of astaxanthin extract was evaluated in experimental mice. Astaxanthin administration was found to enhance the proliferation of spleen cells and bone marrow cells. Esterase activity was found to be enhanced in bone marrow cells indicating increased maturation of cells of lymophoid linkage. Astaxanthin also enhanced number of antibody forming cells and circulating antibody titre. Thus astaxanthin exhibits strong immunomodulating properties. A significant reduction in the viability of ascites tumour cells DLA in vitro was noted in the current study. The % viability was reduced to 4.34 % at a concentration of 15μg astaxanthin/ml. The cytotoxic action of astaxanthin against DLA may be through induction of apoptosis or through a different pathway. Antitumour activity of astaxanthin was studied by ascite and solid tumour models in mice. An increase in life span of about 67 % was noted in DLA bearing mice administered with astaxanthin at 5 mg/kg body weight. The tumour volume and tumour weight were significantly lower in mice injected with 5 mg/kg body weight astaxanthin. In vitro studies revealed that astaxanthin from shrimp shell waste of Aristeus alcocki inhibited the proliferation of cervical cancer cells HeLa in a dose dependent manner.ThesisItem Open Access GENETIC STUDIES IN F3 GENERATION OF THREE CROSSES IN GREEN GRAM (Vigna radiata (L.) Wilczek)(Acharya N.G. Ranga Agricultural University, 2017) SINDHU, S; SHANTHI PRIYA, MThe present investigation entitled “Genetic studies in F3 generation of three crosses in green gram (Vigna radiata (L.) Wilczek)” was carried out during kharif, 2016 to study the variability, heritability, genetic advance as per cent of mean, character association for different yield related characters and their direct and indirect effects on seed yield and transgressive segregants for yield, harvest index and water use efficiency related traits. The experimental material used in this study consists of four parents viz., MGG-347, MGG-351, LGG-460 and LGG-528 and three F3 populations of MGG-347 x MGG-351, MGG-351 x LGG-460 and LGG-460 x LGG-528. The experiment was laid out in compact family block design with three replications at dry land farm, S.V. Agricultural College, Tirupati. The analysis of variance between families revealed that the mean squares due to crosses were significant for the characters viz., number of pods per plant, specific leaf area at 35 DAS, specific leaf area at 50 DAS and SCMR at 50 DAS. The analysis of variance among progenies within each family indicated significant differences among progeny means for all the characters studied in all the crosses except for number of primary branches per plant, number of pods per plant, number of pods per cluster, harvest index and seeds per pod in cross MGG-347 x MGG-351; number of primary branches per plant and harvest index in cross LGG-460 x LGG-528 and hundred seed weight in all the three crosses viz., MGG-347 x MGG-351, MGG-351 x LGG-460 and LGG-460 x LGG-528. Estimates of phenotypic coefficient of variation (PCV) were higher than genotypic coefficient of variation (GCV) for all the characters. High GCV estimates and high heritability coupled with high genetic advance as per cent of mean were observed for number of clusters per plant, number of pods per plant, specific leaf area at 35 DAS, pod yield and seed yield in the two crosses, MGG-351 x LGG-460 and LGG-460 x LGG-528 suggesting the involvement of additive gene action for these characters in these crosses. Hence, direct selection for these traits in these crosses would result in further improvement of seed yield along with water use efficiency in green gram. High heritability and genetic advance as per cent of mean were observed for SLA at 35 DAS and 50 DAS in all the three crosses, suggesting that these characters were governed by additive gene action and simple selections can be utilized to improve water use efficiency in these crosses. Based on association analysis, plant height, number of cluster per plant, number of pods per plant, number of pods per cluster, number of seeds per pod, harvest index and pod yield might be selected for improvement in seed yield in positive direction in all the three crosses. Path analysis revealed that in all the three crosses, highest positive direct effect on seed yield was exerted by pod yield followed by harvest index. Hence, these traits might be considered as selection criteria for breeding high yielding lines. A good number of transgressive segregants over better parent were propelled for number of primary branches per plant, number of pods per plant and SCMR at 35 DAS in the cross MGG-347 x MGG-351; plant height, number of pods per cluster, number of seeds per pod, hundred seed weight and SLA at 35 DAS in the cross MGG-351 x LGG-460; number of clusters per plant, pod length, SLA at 50 DAS, SCMR at 50 DAS, harvest index, pod yield and seed yield in the cross LGG-460 x LGG-528. Hence, forwarding these transgressive segregants to further generation, contributes to the development of most promising genotype which is better than both the parents. Among all the three crosses, LGG-460 x LGG-528 cross can be used to breed high yielding lines and MGG-347 x MGG-351 cross can be used to breed high water use efficiency lines