Use of microalgal biomass as functional ingredient for preparation of cereal based extrudates: impact of processing on amino acid concentrations and colour degradation kinetics

Abstract Suitability of developing Spirulina incorporated cereal based low cost nutritious extrudates was analysed against extrusion processing parameters. Most significant extrusion processing parameters considered for present study were feed moisture (20-25%), die temperature (100-120 °C) and screw speed (50-100 rpm). Different extrusion conditions were used to obtain most acceptable rice: Spirulina blend extrudates. In present study before extrusion processing different additives (citric acid and sodium bicarbonate) were added in rice: Spirulina blend and checked its effect on colour degradation kinetics at varied packaging and storage conditions. Higher screw speed (100 rpm) indicating less residence time of feed material inside the barrel resulted in higher colour retention of rice: Spirulina (97:03) blend extrudates. Kinetics for rice: Spirulina (97:03) blend extrudates indicates faster rate of colour degradation in terms of lightness (half-life of 4 days) when packed in metalized polyethylene at 50°C with 65% relative humidity. Increased concentration of Spirulina (1-3%) in raw formulations resulted in increase in concentration of all amino acids. Impact of extrusion processing has shown non-significant (p ≤ 0.05) effect on amino acid concentrations of rice: Spirulina blend extrudates. Also, all the spirulina added samples showed good consumer acceptability with the score of 6.7

Beneficial effects of Morus alba and Azadirachta indica leaf extract based combination cream in scalding type burn injury in rats.

Background: The present study was designed to evaluate the potential of a combination cream of aqueous extracts of leaves of Morus alba (MA) and Azadirachta indica (AI) in scalding type burn wound injury in rats. Methods: Plant material was successively extracted and aqueous extracts were selected. Three extract based cream formulations viz. 20% w/w (MA), 20% w/w (AI), and combination cream containing 10%+10% w/w (MA+AI) were prepared. Cream base and standard anti-burn cream containing silver sulfadiazine were also used for comparison. Scalding type burn was given by pouring water at 90°C on a shaved dorsal area of 20 mm2. Deep second-degree burn injury was produced which was evaluated for next 21 days for a percentage of wound contraction and period of epithelialization. On 21st day, animals were sacrificed and histopathological slides were prepared using hematoxylin-eosin staining. Burned tissue was also screened for levels of oxidative stress using thiobarbituric acid reactive substances (TBARS) and glutathione (GSH) estimation. Results: There was a significant increase in the percentage of wound contraction and significant decrease in period of epithelialization in MA, AI, and MA+AI combination cream treated group as compared with control group. However, most significant results were obtained with MA+AI combination cream. Histologically, MA+AI cream treatment resulted in almost complete re-epithelialization and restructuring of the wound tissue. There was a significant rise in TBARS and decrease in GSH levels in burn injury group which was reversed to a major extent by the application of combination cream. Conclusions: The results indicate toward the possible role of free radical scavenging potential of extracts in the Burn wound tissue healing.

DPPH radical scavenging activity and contents of H2O2, malondialdehyde and proline in determining salinity tolerance in chickpea seedlings.

The involvement of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and contents of H2O2, malondialdehyde (MDA) and proline was investigated in determining salinity tolerance among seedlings of thirty chickpea (Cicer arietinum L.) genotypes having different pedigrees. Chickpea genotypes, including cultivars and advanced lines were grown for 7 days under control and salt stress (50 mM NaCl) conditions. The genotypes showed differential response to salt stress in terms of growth, DPPH radical scavenging activity and contents of H2O2, MDA and proline in seedlings. On the basis of seedling growth, the genotypes having better performance under stress conditions had reduced levels of H2O2 and MDA contents, but increased levels of proline and DPPH radical scavenging activity. Stress tolerance index for these parameters was also determined. Agglomerative hierarchal clustering by Pearson correlation coefficient grouped the genotypes into two major clusters — MC I and MC II. MC II and A1-1 sub-cluster of MC-I comprised mainly of genotypes that showed higher stress resistance levels for the respective parameters in comparison to genotypes in other sub-clusters. Thus, it is possible to identify salt-tolerant genotypes on the basis of above parameters without a field trial.

Differential antioxidative response of tolerant and sensitive maize (Zea mays L.) genotypes to drought stress at reproductive stage.

The role of oxidative stress management was evaluated in two maize (Zea mays L.) genotypes — Parkash (drought-resistant) and Paras (drought-sensitive), subjected to drought stress during reproductive stage. Alterations in their antioxidant pools — glutathione (GSH) and ascorbic acid (AsA) combined with activities of enzymes glutathione reductase (GR), ascorbate peroxidase (APX), peroxidase (POX) and catalase (CAT) involved in defense against oxidative stress and stress parameters, namely chlorophyll (Chl), hydrogen peroxide (H2O2) and malondialdehyde (MDA) were investigated in flag leaves from silk emergence till maturity. The drought caused transient increase in GR, APX, POX and CAT activities in drought-tolerant genotype (Parkash) which decreased at later stages with the extended period of drought stress. However, in Paras, drought stress caused decrease in activities of GR and CAT from initial period of stress till the end of experiment, except for POX which showed slight increase in activity. A significant increase in GSH content was observed in Parkash till 35 days after silking (DAS), whereas in Paras, GSH content remained lower than irrigated till maturity. Parkash which had higher AsA and Chl contents, also showed lower H2O2 and MDA levels than Paras under drought stress conditions. However, at the later stages, decline in antioxidant enzyme activities in Parkash due to severe drought stress led to enhanced membrane damage, as revealed by the accumulation of MDA. Our data indicated that significant activation of antioxidant system in Parkash might be responsible for its drought-tolerant behavior under drought stress and helped it to cope with the stress up to a definite period. Thus, the results indicate that antioxidant status and lipid peroxidation in flag leaves can be used as indices of drought tolerance in maize plants and also as potential biochemical targets for the crop improvement programmes to develop drought-tolerant cultivars.

High acid invertase activity for a prolonged period in developing seeds/podwall of wild chickpea is detrimental to seed filling.

In the present study factors responsible for low seed biomass in wild Cicer species has been investigated. Cicer judaicum and chickpea cultivar PBG-1 were investigated to compare activities of some enzymes involved in carbon metabolism in podwall and seeds during crop development. Seed filling duration in wild species was about 15 days shorter than that of cultivated varieties due to rapid loss of moisture content and hence resulted in earlier maturity and reduced seed biomass. Longer seed filling duration appeared to be an important factor responsible for greater biomass of chickpea seeds. Because of absence of phosphoenol pyruvate carboxylase from 25-35 days after flowering and low sucrose synthase activities, the podwall of C. judaicum is not in a position to contribute significantly to the sink filling capacity of seeds. High acid invertase, low sucrose synthase activities during seed storage phase cause detrimental effect on seed filling and resulting in highly reduced sink strength and productivity of wild species. Successful transfer of stress tolerance from wild Cicer species to chickpea cultivars need to prevent the transfer of these observed unfavourable biochemical factors so that the productivity of chickpea crop remains unaffected during utilization of wild Cicer species in chickpea improvement.

Potential of antioxidant enzymes in depicting drought tolerance of wheat (Triticum aestivum L.).

In an effort to determine the biochemical markers for identifying genotypes before sowing for drought tolerance, changes in activities of antioxidant enzymes were determined in the seedlings of five drought-tolerant and five drought-sensitive wheat (Triticum aestivum L.) genotypes, each with different genetic background growing under normal and water deficit conditions induced by 6% mannitol. In comparison with non-stressed seedlings, the catalase (CAT) activity was upregulated by more than 50% in the roots of water-stressed seedlings in drought-tolerant genotypes. Water deficit stress also led to the upregulation of ascorbate peroxidase (APX) in the endosperms and glutathione reductase (GR), CAT and peroxidase (POD) in the shoots of stressed seedlings in drought-tolerant genotypes. Superoxide dismutase (SOD) activity was very low in roots and shoots and showed non-significant increase under water-stress in tolerant genotypes. Out of five specified enzyme activities (CAT in roots and shoots, APX in endosperms, GR and POD in shoots), if any three are upregulated in the specified tissues under water deficit conditions, the genotype is likely to be drought-tolerant. Wheat seedlings with low GR and APX activities and high POD activity in shoots with a low ratio of GR activity of shoot to root of non-stressed seedlings are likely to perform better under rainfed conditions. The observed data showed that status of antioxidant enzymes could provide a meaningful tool for depicting drought tolerance of a wheat genotype.

Response of antioxidative and ethanolic fermentation enzymes in maize seedlings of tolerant and sensitive genotypes under short term waterlogging.

Fifteen days old seedlings of waterlogging tolerant (Parkash) and sensitive (Paras) maize genotypes were subjected to short term waterlogging (18 h) under field conditions. Activities of various antioxidative and anaerobic metabolism enzymes were investigated in leaf and root tissues. Superoxide dismutase (SOD) activity increased in leaf tissue while glutathione reductase (GR) activity was enhanced in leaf as well as root in both the genotypes. However, tolerant genotype had better induction capability of SOD and GR in roots in comparison with sensitive genotype. Catalase activity increased in roots of both genotypes. Waterlogging caused strong induction in alcohol dehydrogenase activity in the roots of Paras and Parkash under stress conditions. Aldehyde dehydrogenase activity was significantly increased only in roots of Parkash in response to waterlogging. In comparison with sensitive genotype, the tolerant genotype had low H2O2 and malondialdehyde content in roots under stress conditions. The present studies suggested that tolerant genotype had a greater protective ability due to higher induced activities of antioxidant and ethanolic fermentation systems than Paras.

Role of antioxidant and anaerobic metabolism enzymes in providing tolerance to maize (Zea mays L.) seedlings against waterlogging.

The present investigation was undertaken to identify the possible mode of mechanism that could provide tolerance to maize (Zea mays L.) seedlings under waterlogging. Using cup method, a number of maize genotypes were screened on the basis of survival of the seedlings kept under waterlogging. Two tolerant (LM5 and Parkash) and three susceptible (PMH2, JH3459 and LM14) genotypes were selected for the present study. Activities of antioxidant and ethanolic fermentation enzymes and content of hydrogen peroxide (H2O2), glutathione and ascorbic acid were determined in roots of these genotypes after 72 h of waterlogging. Waterlogging treatment caused decline in activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in all the genotypes. However, only susceptible genotypes showed slight increase in glutathione reductase (GR) activity. Significant reduction in APX/GR ratio in susceptible genotypes might be the cause of their susceptibility to waterlogging. The tolerant seedlings had higher GR activity than susceptible genotypes under unstressed conditions. Stress led to decrease in H2O2 and increase in glutathione content of both tolerant and susceptible genotypes, but only tolerant genotypes exhibited increase in ascorbic acid under waterlogging conditions. In the tolerant genotypes, all the enzymes of anaerobic metabolism viz. alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and pyruvate decarboxylase (PDC) were upregulated under waterlogging, whereas in susceptible genotypes, only ADH was upregulated, suggesting that efficient upregulation of entire anaerobic metabolic machinery is essential for providing tolerance against waterlogging. The study provides a possible mechanism for waterlogging tolerance in maize.

Evaluation of oxidative stress tolerance in maize (Zea mays L.) seedlings in response to drought.

Seedlings of selected six genotypes of maize (Zea mays L.) differing in their drought sensitivity (LM5 and Parkash drought-tolerant and PMH2, JH3459, Paras and LM14 as drought-sensitive) were exposed to 72 h drought stress at two leaf stage. Alterations in their antioxidant pools combined with activities of enzymes involved in defense against oxidative stress were investigated in leaves. Activities of some reactive oxygen species (ROS)-scavenging enzymes, catalase (CAT) and ascorbate peroxidase (APX) were enhanced in tolerant genotypes in response to drought stress. Superoxide dismutase (SOD) activity was significantly decreased in sensitive genotypes, but remained unchanged in tolerant genotypes under stress. Peroxidase (POX) activity was significantly induced in tolerant, as well as sensitive genotypes. Imposition of stress led to increase in H2O2 and malondialdehyde (MDA, a marker for lipid peroxidation) content in sensitive genotypes, while in tolerant genotypes no change was observed. Significant increase in glutathione content was observed in sensitive genotypes. Ascorbic acid pool was induced in both tolerant and sensitive genotypes, but induction was more pronounced in tolerant genotypes. Significant activation of antioxidative defence mechanisms correlated with drought-induced oxidative stress tolerance was the characteristic of the drought tolerant genotypes. These studies provide a mechanism for drought tolerance in maize seedlings.

Emergence of VRE and their antimicrobial sensitivity pattern in a tertiary care teaching hospital

During the last few years enterococci have emerged as an important cause of nosocomial and community acquired infection. They have acquired resistance to commonly used antibiotics including glycopeptides posing challenge to therapeutic options. The aim of this study was to investigate the prevalence and sensitivity of VRE to newer drugs. A total of 250 strains of E. faecalis were isolated using conventional scheme of Facklam and Collins. High level aminoglycoside resistance (HLAR) was detected by disc diffusion method using 120 ìg gentamicin disc and confirmed by agar dilution screen method. Screening for vancomycin resistance was done by disc diffusion and the agar screen method, and was further confirmed by broth dilution method for minimum inhibitory concentration (MIC). The strains which were resistant to vancomycin were further tested for sensitivity to newer and commonly available antibiotics. Maximum number of enterococcal isolates were recovered from urine (32.8 por cento) followed by blood (25.6 por cento) and pus (18.4 por cento). Penicillin (83.6 por cento) and cotrimoxazole (77.9 por cento) were found to be least effective drugs against the E. faecalis whereas; cefuroxime (76.8 por cento) and vancomycin (98 por cento) were most effective drugs in vitro. About two percent isolates of enterococci were resistant to vancomycin. All the VRE isolates were sensitive to quinupristin/dalfopristin. Linezolid and chloramphenicols were the two other in vitro effective drugs with 80 por cento sensitive isolates. MIC of all the VRE isolates was found to be in range of 64-512ìg/mL. So, quinupristin/dalfopristin can be used for infections caused by VRE. Continuous surveillance is necessary to detect early outbreak, and spread of VRE.

Effect of water deficit on carbohydrate status and enzymes of carbohydrate metabolism in seedlings of wheat cultivars.

The effect of water deficit on carbohydrate status and enzymes of carbohydrate metabolism (alpha and beta amylases, sucrose phosphate synthase, sucrose synthase, acid and alkaline invertases) in wheat (Triticum aestivum L.) was investigated in the seedlings of drought-sensitive (PBW 343) and drought-tolerant (C 306) cultivars. The water deficit was induced by adding 6% mannitol (water potential -0.815 Mpa) in the growth medium. The water deficit reduced starch content in the shoots of tolerant seedlings as compared to the sensitive ones, but increased sucrose content in the shoots and roots of tolerant seedlings, indicating their protective role during stress conditions. It also decreased the alpha-amylase activity in the endosperm of seedlings of both the cultivars, but increased alpha and beta amylase activities in the shoots of tolerant ones. Sucrose phosphate synthase (SPS) activity showed a significant increase at 6 days of seedling growth (DSG) in the shoots of stressed seedlings of tolerant cultivar. However, SPS activity in the roots of stressed seedlings of sensitive cultivar was very low at 4 DSG and appeared significantly only at day 6. Sucrose synthase (SS) activity was lower in the shoots and roots of stressed seedlings of tolerant cultivar than sensitive ones at early stage of seedling growth. Higher acid invertase activity in the shoots of seedlings of tolerant cultivar appeared to be a unique characteristic of this cultivar for stress tolerance. Alkaline invertase activity, although affected under water deficit conditions, but was too low as compared to acid invertase activity to cause any significant affect on sucrose hydrolysis. In conclusion, higher sucrose content with high SPS and low acid invertase and SS activities in the roots under water deficit conditions could be responsible for drought tolerance of C 306.

Sugar signalling and gene expression in relation to carbohydrate metabolism under abiotic stresses in plants.

Sucrose is required for plant growth and development. The sugar status of plant cells is sensed by sensor proteins. The signal generated by signal transduction cascades, which could involve mitogen-activated protein kinases, protein phosphatases, Ca 2+ and calmodulins, results in appropriate gene expression. A variety of genes are either induced or repressed depending upon the status of soluble sugars. Abiotic stresses to plants result in major alterations in sugar status and hence affect the expression of various genes by down- and up-regulating their expression. Hexokinase-dependent and hexokinase-independent pathways are involved in sugar sensing. Sucrose also acts as a signal molecule as it affects the activity of a proton-sucrose symporter. The sucrose trans-porter acts as a sucrose sensor and is involved in phloem loading. Fructokinase may represent an additional sensor that bypasses hexokinase phosphorylation especially when sucrose synthase is dominant. Mutants isolated on the basis of response of germination and seedling growth to sugars and reporter-based screening protocols are being used to study the response of altered sugar status on gene expression. Common cis-acting elements in sugar signalling pathways have been identified. Transgenic plants with elevated levels of sugars/sugar alcohols like fructans, raffinose series oligosaccharides, trehalose and mannitol are tolerant to different stresses but have usually impaired growth. Efforts need to be made to have transgenic plants in which abiotic stress responsive genes are expressed only at the time of adverse environmental conditions instead of being constitutively synthesized.

Applications of inulin and oligofructose in health and nutrition.

Inulin and oligofructose belong to a class of carbohydrates known as fructans. The main sources of inulin and oligofructose that are used in the food industry are chicory and Jerusalem artichoke. Inulin and oligofructose are considered as functional food ingredients since they affect the physiological and biochemical processes in rats and human beings, resulting in better health and reduction in the risk of many diseases. Experimental studies have shown their use as bifidogenic agents, stimulating the immune system of the body, decreasing the pathogenic bacteria in the intestine, relieving constipation, decreasing the risk of osteoporosis by increasing mineral absorption, especially of calcium, reducing the risk of atherosclerosis by lowering the synthesis of triglycerides and fatty acids in the liver and decreasing their level in serum. These fructans modulate the hormonal level of insulin and glucagon, thereby regulating carbohydrate and lipid metabolism by lowering the blood glucose levels; they are also effective in lowering the blood urea and uric acid levels, thereby maintaining the nitrogen balance. Inulin and oligofructose also reduce the incidence of colon cancer. The biochemical basis of these beneficial effects of inulin and oligofructose have been discussed. Oligofructose are non cariogenic as they are not used by Streptococcus mutans to form acids and insoluble glucans that are the main culprits in dental caries. Because of the large number of health promoting functions of inulin and oligofructose, these have wide applications in various types of foods like confectionery, fruit preparations, milk desserts, yogurt and fresh cheese, baked goods, chocolate, ice cream and sauces. Inulin can also be used for the preparation of fructose syrups.