Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 212

Search results for: In vitro digestibility

212 The Effect of Carboxymethyl Cellulose on the Stability of Emulsions Stabilized by Whey Proteins under Digestion in vitro and in vivo

Authors: D. Leskauskaite, I. Jasutiene, M. Kersiene, E. Malinauskyte, P. Matusevicius

Abstract:

In vitro gastro-duodenal digestion model was used to investigate the changes of emulsions under digestion conditions. Oil in water emulsions stabilized by whey proteins (2%) and stabilized by whey proteins (2%) with addition of carboxymethyl cellulose (0.75%) as gelling agent of continuous phase were prepared at pH7. Both emulsions were destabilized under gastric conditions; however the protective role of carboxymethyl cellulose was indicated by recording delay of fat digestibility of this emulsion. In the presence of carboxymethyl cellulose whey proteins on the interfacial surface of droplets were more resistant to gastric degradation causing limited hydrolysis of fat due to the poor acceptability of lipids for the enzymes. Studies of emulsions using in vivo model supported results from in vitro studies. Lower content of triglycerides in blood serum and higher amount of fecal fat of rats were determined when rats were fed by diet containing emulsion made with whey proteins and carboxymethyl cellulose. 

Keywords: Digestibility, emulsions, lipids, rats.

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211 Evaluation of Protein Digestibility in Canola Meals between Caecectomised and Intact Adult Cockerels

Authors: Ali Nouri Emamzadeh, Akbar Yaghobfar

Abstract:

The experiment was conducted to evaluate digestibility quantities of protein in Canola Meals (CMs) between caecectomised and intact adult Rhode Island Red (RIR) cockerels with using conventional addition method (CAM) for 7 d: a 4-d adaptation and a 3-d experiment period on the basis of a completely randomized design with 4 replicates. Results indicated that caecectomy decreased (P<0.05) apparent and true digestibility quantities of protein for CMs, except for CMs 2 and 3. The mean apparent and true digestibility quantities for all CMs in caecectomised (80.5 and 81.4%, respectively) were (3.1 and 3.3%, respectively) less (P<0.05) than intact cockerels (83.6 and 84.7%, respectively). Therefore, the caecectomy method increases accuracy of the digestibility measurements of protein for this meal in bioassays based on excreta collection in adult cockerels.

Keywords: Adult cockerels, caecectomy, canola meals, proteindigestibility.

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210 Biochemical Characteristics of Sorghum Flour Fermented and/or Supplemented with Chickpea Flour

Authors: Omima E. Fadlallah, Abdullahi H. El Tinay, Elfadil E. Babiker

Abstract:

Sorghum flour was supplemented with 15 and 30% chickpea flour. Sorghum flour and the supplement were fermented at 35 oC for 0, 8, 16, and 24 h. Changes in pH, titrable acidity, total soluble solids, protein content, in vitro protein digestibility and amino acid composition were investigated during fermentation and/or after supplementation of sorghum flour with chickpea. The pH of the fermenting material decreased sharply with a concomitant increase in the titrable acidity. The total soluble solids remained unchanged with progressive fermentation time. The protein content of sorghum cultivar was found to be 9.27 and that of chickpea was 22.47%. The protein content of sorghum cultivar after supplementation with15 and 30% chickpea was significantly (P ≤ 0.05) increased to 11.78 and 14.55%, respectively. The protein digestibility also increased after fermentation from 13.35 to 30.59 and 40.56% for the supplements, respectively. Further increment in protein content and digestibility was observed when supplemented and unsupplemented samples were fermented for different periods of time. Cooking of fermented samples was found to increase the protein content slightly and decreased digestibility for both supplements. Amino acid content of fermented and fermented and cooked supplements was determined. Supplementation was found to increase the lysine and therionine content. Cooking following fermentation decreased lysine, isoleucine, valine and sulfur containg amino acids.

Keywords: Amino acid, Chickpea, Cooking, Fermentation, protein, Sorghum.

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209 Degree of Milling Effects on the Sorghum (Sorghum bicolor) Flours, Physicochemical Properties and Kinetics of Starch Digestion

Authors: Brou K., Guéhi T., Konan A. G., Gbakayoro J. B., Gnakri D.

Abstract:

Two types of crushing were applied to grains of red sorghum: manual crushing using a mortar and pestle of kitchen and mechanical crushing using a hammer mill. The flours obtained at the end of these various crushing were filtered and subdivided in different fractions according to the diameters of the mesh of the sieves (0.16mm; 0.25mm; 0.315mm; 0.4mm, and 0.63mm…). Some physical, chemical and nutritional traits of these flours were evaluated using Association of Official Analytical Chemists (AOAC). In vitro digestibility of these flours was also studied with freezing of flour 1% like substrate and α-amylase from B. licheniformis (E.C.3.2.1.1; Megazyme, Wicklow, Ireland). The results revealed that the batches of flours which have the finest diameters as 0.16mm; 0.25mm are the richest one in nutrients and are also the most digestible. Also mechanical crushing is the best mean to obtain significant amount of flours. In conclusion, the type of crushing and the size of the particles have an impact on the final concentration of some nutrients of the flours obtained. Indeed, the finest particles (0.16mm – 0.25mm 0.315mm) obtained after sifting of the flours are more nutritive and have a better digestibility than others size. So the finest particles could be advised for management of cereals namely the sorghum for the production of the infantile foods.

Keywords: Nutrients, digestibility, crush, flour, milling, granulometry.

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208 Digestibility in Yankasa Rams Fed Brachiaria ruziziensis – Centrosema pascuorum Hay Mixtures with Concentrate

Authors: Ibrahim Sani, J. T. Amodu, M. R. Hassan, R. J. Tanko, N. Adamu

Abstract:

This study investigated the digestibility of Brachiaria ruziziensis and Centrosema pascuorum hay mixtures at varying proportions in Yankasa rams. Twelve Yankasa rams with average initial weight 10.25 ± 0.1 kg were assigned to three dietary treatments of B. ruziziensis and C. pascuorum hay at different mixtures (75BR:25CP, 50BR:50CP and 25BR:75CP, respectively) in a Completely Randomized Design (CRD) for a period of 14 days. Concentrate diet was given to the experimental animals as supplement at fixed proportion, while the forage mixture (basal diet) was fed at 3% body weight. Animals on 50BR:50CP had better nutrient digestibility (crude protein, acid and neutral detergent fibre, ether extract and nitrogen free extract) than other treatment diets, except in dry matter digestibility (87.35%) which compared with 87.54% obtained in 25BR:75CP treatment diet and also organic matter digestibility. All parameters taken on nitrogen balance with the exception of nitrogen retained were significantly higher (P < 0.05) in animals fed 25BR:75CP diet, but were statistically similar with values obtained for animals on 50BR:50CP diet. From results obtained in this study, it is concluded that mixture of 25%BR75%CP gave the best nutrient digestibility and nitrogen balance in Yankasa rams. It is therefore recommended that B. ruziziensis and C. pascuorum should be fed at 50:50 mixture ratio for enhanced animal growth and performance in Nigeria.

Keywords: B. ruziziensis, C. pascuorum, digestibility, rams, Yankasa.

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207 Evaluation of Hazelnut Hulls as an Alternative Forage Resource for Ruminant Animals

Authors: N. Cetinkaya, Y. S. Kuleyin

Abstract:

The aim of this study was to estimate the digestibility of the fruit internal skin of different varieties of hazelnuts to propose hazelnut fruit skin as an alternative feed source as roughage in ruminant nutrition. In 2015, the fruit internal skins of three different varieties of round hazelnuts (RH), pointed hazelnuts (PH) and almond hazelnuts (AH) were obtained from hazelnut processing factory then their crude nutrients analysis were carried out. Organic matter digestibility (OMD) and metabolisable energy (ME) values of hazelnut fruit skins were estimated from gas measured by in vitro gas production method. Their antioxidant activities were determined by spectrophotometric method. Crude nutrient values of three different varieties were; organic matter (OM): 87.83, 87.81 and 87.78%), crude protein (CP): 5.97, 5.93 and 5.89%, neutral detergent fiber (NDF): 30.30, 30.29 and 30.29%, acid detergent fiber (ADF): 48.68, 48.67 and 48.66% and acid detergent lignin (ADL): 25.43, 25.43 and 25.39% respectively. OMD from 24 h incubation time of RH, PH and AH were 22.04, 22.46 and 22.74%; MEGP values were 3.69, 3.75 and 3.79 MJ/kg DM; and antioxidant activity values were 94.60, 94.54 and 94.52 IC 50 mg/mL respectively. The fruit internal skin of different varieties of hazelnuts may be considered as an alternative roughage for ruminant nutrition regarding to their crude and digestible nutritive values. Moreover, hazelnut fruit skin has a rich antioxidant content so it may be used as a feed additive for both ruminant and non-ruminant animals.

Keywords: Antioxidant activity, hazelnut fruit skin, metabolizable energy, organic matter digestibility.

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206 Chemical and Biological Properties of Local Cowpea Seed Protein Grown in Gizan Region

Authors: Abdelatief S. H. El-Jasser

Abstract:

The aim of the present study was to investigate the chemical and biological properties of local cowpea seed protein cultivated in Gizan region. The results showed that the cowpea and its products contain high level of protein (22.9-77.6%), high carbohydrates (9.4-64.3%) and low fats (0.1-0.3%). The trypsin and chymotrypsin activities were found to be 32.2 and 15.2 units, respectively. These activities were not affected in both defatted and protein concentrate whereas they were significantly reduced in isolated protein and cooked samples. The phytate content of cooked and concentrated cowpea samples varied from 0.25% -0.32%, respectively. Tannin content was found to be 0.4% and 0.23% for cooked and raw samples, respectively. The in vitro protein digestibility was very high in cowpea seeds (75.04-78.76%). The biological evaluation using rats showed that the group fed with animal feed containing casein gain more weight than those fed with that containing cowpea. However, the group fed with cooked cowpea gain more weight than those fed with uncooked cowpea. On the other hand, in vivo digestion showed high value (98.33%) among the group consumed casein compared to other groups those consumed cowpea contains feed. This could be attributed to low antinutritional factors in casein contains feed compared to those of cowpea contains feed because cooking significantly increased the digestion rate (80.8% to 83.5%) of cowpea contains feed. Furthermore, the biological evaluation was high (91.67%) of casein containing feed compared to that of cowpea containing feed (80.83%-87.5%). The net protein utilization (NPU) was higher (89.67%) in the group fed with casein containing feed than that of cowpea containing feed (56.33%-69.67%).

Keywords: Biological properties, Cowpea seed protein, Antinutritional factors, In vitro digestibility

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205 The effects of Garlic Oil (Allium sativa), Turmeric Powder (Curcuma longa Linn) and Monensin on Total Apparent Digestibility of Nutrients in Baloochi Lambs

Authors: Ahmad Khalesizadeh, Alireza Vakili, Mohsen Danesh Mesgaran, Reza Valizadeh

Abstract:

The objective of this study was to determine the effects of garlic oil (Allium sativa), turmeric powder (Curcuma longa Linn) and Monensin on Total apparent digestibility of nutrients in Baloochi lambs. The experiment was designed as a 4 x 4 Latin square using 4 ruminally baloochi lambs with 4 treatments in four 28-d periods. Treatments were control (no additive), garlic oil (0. 4 g/d), monensin (0. 2 g/d) and turmeric powder (20 g/d). Total apparent digestibility's (% of intake) of organic matter (OM), dry matter (DM), crud protein (CP), ether extract(EE), non fiber carbohydrate (NFC), acid detergent fiber (ADF) and neutral detergent fiber (NDF) in the total tract were not influenced by addition of either additives.

Keywords: apparent digestibility, essential oil, garlic oil, monensin, turmeric

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204 The Effect of Granule Size on the Digestibility of Wheat Starch Using an in vitro Model

Authors: Mee-Lin Lim Chai Teo, Darryl M. Small

Abstract:

Wheat has a bimodal starch granule population and the dependency of the rate of enzymatic hydrolysis on particle size has been investigated. Ungelatinised wheaten starch granules were separated into two populations by sedimentation and decantation. Particle size was analysed by laser diffraction and morphological characteristics were viewed using SEM. The sedimentation technique though lengthy, gave satisfactory separation of the granules. Samples (<10μm, >10μm and original) were digested with a-amylase using a dialysis model. Granules of <10μm showed significantly higher rate of reducing sugar release than those >10μm (p<0.05). In contrast, the rate was not significantly different between the original sample and granules >10μm. Moreover, the digestion rate was dependent on particle size whereby smaller granules produced higher rate of release. The methodology and results reported here can be used as a basis for further evaluations designed to delay the release of glucose during the digestion of native starches.

Keywords: in vitro Digestion, a-amylase, wheat starch, granule size.

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203 Influence of Supplemental Glutamine on Nutrient Digestibility and Utilization, Small Intestinal Morphology and Gastrointestinal Tract and Immune Organ Developments of Broiler Chickens

Authors: Sutisa Khempaka, Supattra Okrathok, Laddawan Hokking, Buntita Thukhanon, Wittawat Molee

Abstract:

This study was conducted to investigate the optimum levels of glutamine (Gln) supplementation in broiler diets. A total of 32 one-day-old male chicks with initial body weight 41.5 g were segregated into 4 groups (8 chicks per group) and subsequently distributed to individual cages. Feed and water were provided ad libitum for 21 days. Four dietary treatments were as follows: control and supplemented Gln at 1, 2 and 3%, respectively. The results found that the addition Gln had no negative effects on dry matter, organic matter, ash digestibility or nitrogen retention. Birds fed with 1% Gln had significantly higher villi wide and villi height : crypt depth ratio in duodenum than the control chicks and 2 and 3% Gln chicks. It is suggested that the addition of Gln at 1% indicated a beneficial effect on improving small intestinal morphology, in addition Gln may stimulate immune organ development of broiler chickens.

Keywords: broiler chicken, digestibility, gastrointestinal tract glutamine, glutamine

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202 In silico Simulations for DNA Shuffling Experiments

Authors: Luciana Montera

Abstract:

DNA shuffling is a powerful method used for in vitro evolute molecules with specific functions and has application in areas such as, for example, pharmaceutical, medical and agricultural research. The success of such experiments is dependent on a variety of parameters and conditions that, sometimes, can not be properly pre-established. Here, two computational models predicting DNA shuffling results is presented and their use and results are evaluated against an empirical experiment. The in silico and in vitro results show agreement indicating the importance of these two models and motivating the study and development of new models.

Keywords: Computer simulation, DNA shuffling, in silico andin vitro comparison.

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201 Cultivation of Thymus by In Vitro And Hydroponics Combined Method

Authors: E. Sargsyan, A. Vardanyan, L. Ghalachyan, S. Bulgadaryan

Abstract:

Our results showed that for the growth of qualitative seedling and vegetative raw material of ðó. marschallianus Willd. and T. serphyllum L. it is more profitable to use the in vitro and hydroponics combined method. In in vitro culture it is possible to do micro-propagation whole year with 98-99% rhizogenesis. 30000 micro-plants were obtained from one explant during 9 months. Hydroponic conditions provide the necessary microclimate for microplants where the survival rate without acclimatization was 93.3%. The essential oil content in hydroponic dry herb of both species in vegetative and blossom phase was 1.3% whereas in wild plants it was 1.2%, the content of extractive substances and vitamin C also exceeded wild plants. Our biochemical and radiochemical investigations indicated that the medicinal raw materials obtained from hydroponic and wild plants of Thymus species correspond to the demands of SPh XI, and the content of artificial radionuclides does not exceed the MACL.

Keywords: Hydroponics, In vitro, Micro-propagation, Thymus

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200 Endothelial Specificity of ICAM2, Flt-1, and Tie2 Promoters In Vitro and In Vivo

Authors: Jing Lei, Yoram Vodovotz, Timothy R. Billiar

Abstract:

To identify an endothelial cell-specific promoter suitable for vascular-specific targeting, we tested five promoters in vitro--Tie2SE, Tie2LE, ICAM2, Flt-1 and vWF--for promoter activity and specificity in endothelial cells, smooth muscle cells and non-vascular resident cells as well as tissues. These promoters, except for vWF, exhibited good endothelial activity and specificity in vitro. In a syngenic heart transplantation model, the ICAM2 promoter was variably functional in coronary endothelial cells of donor hearts. Thus, the ICAM2, Flt-1, Tie2SE and Tie2LE promoters hold promise for endothelial-specific targeting, but in vitro expression may not predict in vivo expression.

Keywords: vascular-specific targeting, endothelial cell-specificpromoter, endothelial specificity.

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199 In vitro Antioxidant Activity of Two Selected Herbal Medicines

Authors: S. Vinotha, I. Thabrew, S. Sri Ranjani

Abstract:

Hot aqueous and methanol extracts of the two selected herbal medicines such are Vellarugu chooranam (V.C) and Amukkirai Chooranam (A.C) were examined for total phenolic and flavonoid contents and in vitro antioxidant activity using four different methods. The total phenolic and flavonoid contents in methanol extract of V.C were found to be higher (44.41±1.26mg GAE/g; 174.44±9.32mg QE/g) than in the methanol extract of A.C (20.56±0.67mg GAE/g; 7.21±0.85mg QE/g). Hot methanol and aqueous extracts of both medicines showed low antioxidant activity in DPPH, ABTS, and FRAP methods and Iron chelating activity not found at highest possible concentration. V.C contains higher concentrations of total phenolic and flavonoid contents than A.C and can also exert greater antioxidant activity than A.C, although the activities demonstrated were lower than the positive control Trolox. The in vitro antioxidant activity was not related with the total phenolic and flavonoid contents of the methanol and aqueous extracts of both herbal medicines (A.C and V.C).

Keywords: Activity, Different extracts, Herbal medicine, in vitro antioxidant.

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198 Effect of Recombinant Human Follicle Stimulating Hormone on Meiotic Competence of In Vitro Grown Nili Ravi Buffalo Oocytes

Authors: Muhammad Ijaz Khan, Samina Jalali, Beenish Shahid, S. A. Shami, Muhammad Ikramullah

Abstract:

In the present study, the response of Nili Ravi buffalo oocytes to recombinant human follicle stimulating hormone (rhFSH) (Organon) on meiotic maturation in vitro was examined. Oocytes were matured in vitro in medium containing either 0 or 0.05 IU/ ml rhFSH and the stage of nuclear maturation recorded after 24 hours. The percentage of oocytes in the control group undergoing germinal vesicle breakdown (GVBD) observed after 24 hours of culture was 29 % whereas as in rhFSH group the percentage was 10 % were at this stage (P< 0.001).Thus in the presence of rhFSH, a significantly greater number of oocytes had progressed to the more advanced stages of nuclear maturation. Indeed, the maturation of GV (Germinal Vesicle) stage oocytes to the metaphase II (M II) stage after 24 hours was significantly (P< 0.0001) increased by the addition of rhFSH (82 % VS 47 %). The percentage of degenerated oocytes after 24 hours of culture was 24 % in control group, whereas in rhFSH group the percentage was 8 % after 24 hours. Degeneration of the oocytes after 24 hours was not significantly (P = 0. 9361) decreased.

Keywords: Buffalo, in vitro, oocytes, recombinant FSH.

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197 Changes of in vitro Cytokine Production induced by δ-Lactams

Authors: Y. Baba hamed, A. Medjdoub, H. Merzouk, M. Narce

Abstract:

The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Keywords: Cytokines, δ-Lactams, In vitro Lymphocyte Proliferation, Superoxide Anion

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196 Novel Solid Lipid Nanoparticles for Oral Delivery of Oxyresveratrol: Effect of the Formulation Parameters on the Physicochemical Properties and in vitro Release

Authors: Y. Sangsen, K. Likhitwitayawuid, B. Sritularak, K. Wiwattanawongsa, R. Wiwattanapatapee

Abstract:

Novel solid lipid nanoparticles (SLNs) were developed to improve oral bioavailability of oxyresveratrol (OXY). The SLNs were prepared by a high speed homogenization technique, at an effective speed and time, using Compritol® 888 ATO (5% w/w) as the solid lipid. The appropriate weight proportions (0.3% w/w) of OXY affected the physicochemical properties of blank SLNs. The effects of surfactant types on the properties of the formulations such as particle size and entrapment efficacy were also investigated. Conclusively, Tween 80 combined with soy lecithin was the most appropriate surfactant to stabilize OXY-loaded SLNs. The mean particle size of the optimized formulation was 134.40 ± 0.57 nm. In vitro drug release study, the selected S2 formulation showed a retarded release profile for OXY with no initial burst release compared to OXY suspension in the simulated gastrointestinal fluids. Therefore, these SLNs could provide a suitable system to develop for the oral OXY delivery.

Keywords: Solid lipid nanoparticles, Physicochemical properties, in vitro drug release, Oxyresveratrol.

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195 In vitro Environmental Factors Controlling Root Morphological Traits of Pineapple (Ananas comosus L. Merr)

Authors: S. Mohajer, R. M. Taha, M. Adel

Abstract:

Developing our knowledge of when pineapple roots grow can lead to improved water, fertilizer applications, and more precise culture management. This paper presents current understanding of morphological traits in pineapple roots, highlighting studies using incubation periods and various solid MS media treated with different sucrose concentrations and pH, which directly assess in vitro environmental factors. Rooting parameters had different optimal sucrose concentrations and incubation periods. All shoots failed to root in medium supplemented with sucrose at 5 g/L and no roots formed within the first 45 days in medium enriched with sucrose at 10 g/L. After 75 days, all shoots rooted in medium enriched with 10 and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L sucrose resulted in the longest and the highest number of roots with 27.3 mm and 4.7, respectively. Root function, such as capacity for P and N uptake, declined rapidly with root length. As a result, the longer the incubation period, the better the rooting responses would be.

Keywords: Environmental factors, in vitro rooting, pineapple, tissue culture.

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194 Effect of Different Media and Mannitol Concentrations on Growth and Development of Vandopsis lissochiloides (Gaudich.) Pfitz. under Slow Growth Conditions

Authors: J. Linjikao, P. Inthima, A. Kongbangkerd

Abstract:

In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m2s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.

Keywords: Preservation, Vandopsis, germplasm, in vitro.

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193 Metoprolol Tartrate-Ethylcellulose Tabletted Microparticles: Development of a Validated Invitro In-vivo Correlation

Authors: Fatima Rasool, Mahmood Ahmad, Ghulam Murtaza, Haji M. S. Khan, Shujaat A. Khan, Sonia Khiljee, Muhammad Qamar-Uz-Zaman

Abstract:

This study describes the methodology for the development of a validated in-vitro in-vivo correlation (IVIVC) for metoprolol tartrate modified release dosage forms with distinctive release rate characteristics. Modified release dosage forms were formulated by microencapsulation of metoprolol tartrate into different amounts of ethylcellulose by non-solvent addition technique. Then in-vitro and in-vivo studies were conducted to develop and validate level A IVIVC for metoprolol tartrate. The values of regression co-efficient (R2-values) for IVIVC of T2 and T3 formulations were not significantly (p<0.05) different from 1 while the values of R2 for IVIVC of T1 and Mepressor® were significantly (p<0.05) different from 1. Internal prediction errors of IVIVC, calculated from observed Area under Curve (AUC) and predicted AUC, were less than 10%. This study successfully presents a valid level A IVIVC for metoprolol tartrate modified dosage forms.

Keywords: Metoprolol tartrate, Dissolution, Bioavailability, Validated in-vitro in-vivo correlation.

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192 Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Authors: Santosh Kumar, Anand Prakash, Kanak Sinha, Anita K Verma

Abstract:

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Keywords: Anti-cancer, Anti-microbial, EAC mouse cell, Tissue culture, Vernonia divergens.

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191 Comparison of the Effects of Continuous Flow Microwave Pre-treatment with Different Intensities on the Anaerobic Digestion of Sewage Sludge for Sustainable Energy Recovery from Sewage Treatment Plant

Authors: D. Hephzibah, P. Kumaran, N. M. Saifuddin

Abstract:

Anaerobic digestion is a well-known technique for sustainable energy recovery from sewage sludge. However, sewage sludge digestion is restricted due to certain factors. Pre-treatment methods have been established in various publications as a promising technique to improve the digestibility of the sewage sludge and to enhance the biogas generated which can be used for energy recovery. In this study, continuous flow microwave (MW) pre-treatment with different intensities were compared by using 5 L semi-continuous digesters at a hydraulic retention time of 27 days. We focused on the effects of MW at different intensities on the sludge solubilization, sludge digestibility, and biogas production of the untreated and MW pre-treated sludge. The MW pre-treatment demonstrated an increase in the ratio of soluble chemical oxygen demand to total chemical oxygen demand (sCOD/tCOD) and volatile fatty acid (VFA) concentration. Besides that, the total volatile solid (TVS) removal efficiency and tCOD removal efficiency also increased during the digestion of the MW pre-treated sewage sludge compared to the untreated sewage sludge. Furthermore, the biogas yield also subsequently increases due to the pre-treatment effect. A higher MW power level and irradiation time generally enhanced the biogas generation which has potential for sustainable energy recovery from sewage treatment plant. However, the net energy balance tabulation shows that the MW pre-treatment leads to negative net energy production.

Keywords: Anaerobic digestion, biogas, microwave pre-treatment, sewage sludge.

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190 In vitro Effects of Salvia officinalis on Bovine Spermatozoa

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

Abstract:

In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: Bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa.

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189 Micropropagation and in vitro Conservation via Slow Growth Techniques of Prunus webbii (Spach) Vierh: An Endangered Plant Species in Albania

Authors: Valbona Sota, Efigjeni Kongjika

Abstract:

Wild almond is a woody species, which is difficult to propagate either generatively by seed or by vegetative methods (grafting or cuttings) and also considered as Endangered (EN) in Albania based on IUCN criteria. As a wild relative of cultivated fruit trees, this species represents a source of genetic variability and can be very important in breeding programs and cultivation. For this reason, it would be of interest to use an effective method of in vitro mid-term conservation, which involves strategies to slow plant growth through physicochemical alterations of in vitro growth conditions. Multiplication of wild almond was carried out using zygotic embryos, as primary explants, with the purpose to develop a successful propagation protocol. Results showed that zygotic embryos can proliferate through direct or indirect organogenesis. During subculture, stage was obtained a great number of new plantlets identical to mother plants derived from the zygotic embryos. All in vitro plantlets obtained from subcultures underwent in vitro conservation by minimal growth in low temperature (4ºC) and darkness. The efficiency of this technique was evaluated for 3, 6, and 10 months of conservation period. Maintenance in these conditions reduced micro cuttings growth. Survival and regeneration rates for each period were evaluated and resulted that the maximal time of conservation without subculture on 4ºC was 10 months, but survival and regeneration rates were significantly reduced, specifically 15.6% and 7.6%. An optimal period of conservation in these conditions can be considered the 5-6 months storage, which can lead to 60-50% of survival and regeneration rates. This protocol may be beneficial for mass propagation, mid-term conservation, and for genetic manipulation of wild almond.

Keywords: Micropropagation, minimal growth, storage, wild almond.

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188 Conditions of the Anaerobic Digestion of Biomass

Authors: N. Boontian

Abstract:

Biological conversion of biomass to methane has received increasing attention in recent years. Grasses have been explored for their potential anaerobic digestion to methane. In this review, extensive literature data have been tabulated and classified. The influences of several parameters on the potential of these feedstocks to produce methane are presented. Lignocellulosic biomass represents a mostly unused source for biogas and ethanol production. Many factors, including lignin content, crystallinity of cellulose, and particle size, limit the digestibility of the hemicellulose and cellulose present in the lignocellulosic biomass. Pretreatments have used to improve the digestibility of the lignocellulosic biomass. Each pretreatment has its own effects on cellulose, hemicellulose and lignin, the three main components of lignocellulosic biomass. Solidstate anaerobic digestion (SS-AD) generally occurs at solid concentrations higher than 15%. In contrast, liquid anaerobic digestion (AD) handles feedstocks with solid concentrations between 0.5% and 15%. Animal manure, sewage sludge, and food waste are generally treated by liquid AD, while organic fractions of municipal solid waste (OFMSW) and lignocellulosic biomass such as crop residues and energy crops can be processed through SS-AD. An increase in operating temperature can improve both the biogas yield and the production efficiency, other practices such as using AD digestate or leachate as an inoculant or decreasing the solid content may increase biogas yield but have negative impact on production efficiency. Focus is placed on substrate pretreatment in anaerobic digestion (AD) as a means of increasing biogas yields using today’s diversified substrate sources.

Keywords: Anaerobic digestion, Lignocellulosic biomass, Methane production, Optimization, Pretreatment.

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187 Manipulation of Probiotics Fermentation of Yogurt by Cinnamon and Licorice: Effects on Yogurt Formation and Inhibition of Helicobacter Pylori Growth in vitro

Authors: S. Behrad, M.Y. Yusof, K. L. Goh, A.S. Baba

Abstract:

Probiotic bacteria especially Lactobacillus spp. and Bifidobacterium exert suppressive effect on Helicobacter pylori. Cinnamon and licorice have been traditionally used for the treatment of gastric ulcer. The objectives of this study were to determine the effects of herbs on yogurt fermentation, the level of probiotic bacteria in yogurt during 28 days storage and the effect of herbal yogurt on the growth of H. pylori in vitro. Cinnamon or licorice was mixed with milk and the mixture was fermented with probiotic bacteria to form herbal-yogurt. Changes of pH and total titratable acids were monitored and the viability of probiotic bacteria was evaluated during and after refrigerated storage. The in vitro inhibition of H. pylori growth was determined using agar diffusion and minimum inhibitory concentration (MIC) method. The presence of herbs did not affect the probiotic population during storage. There were no significant differences in pH and TTA between herbal-yogurts and plain-yogurt during fermentation and storage. Water extract of cinnamon-yogurt showed the highest inhibition effect (13.5mm) on H. pylori growth in comparison with licorice-yogurt (11.2mm). The present findings indicate cinnamon and licorice has bioactive components to decrease the growth of H. pylori.

Keywords: Cinnamon, Helicobacter pylori, Herbal-Yogurt, Licorice, Probiotics

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186 Sterilisation of in vitro Culture Medium of Chrysanthemum by Plant Essential Oils without Autoclaving

Authors: Chockpisit Thepsithar, Aree Thongpukdee, Apichya Daorat

Abstract:

The alternative technique for sterilization of culture medium to replace autoclaving was carried out. For sterilization of culture medium without autoclaving, some commercial pure essential oils, bergamot oil, betel oil, cinnamon oil, lavender oil and turmeric oil, were tested alone or in combinations with some disinfectants, 10% povidone-iodine and 2% iodine + 2.4% potassium iodide. Each essential oil or combination was added to 25-mL Murashige and Skoog (MS) medium before medium was solidified in a 120-mL container, kept for 2 weeks before evaluating sterile conditions. Treated media, supplemented with essential oils, were compared to control medium, autoclaved at 121 degree Celsius for 15 min. In vitro sterile conditions were found 20 – 100% from these treated media compared to 100% sterile condition from autoclaved medium. Treated media obtained 100% sterile conditions were chosen for culturing chrysanthemum shoots. It was found that 10% povidoneiodine in combination with cinnamon oil (3:1) and 2% iodine + 2.4% potassium iodide in combination with lavender oil (1:3) at the concentration of 36 3L/25 mL medium provided the promising growth of shoot explants.

Keywords: Sterilizing agents, essential oils, disinfectants, MS medium, in vitro culture, chrysanthemum, sterilization of medium without autoclaving

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185 Role of Fish Hepatic Aldehyde Oxidase in Oxidative in vitro Metabolism of Phenanthridine Heterocyclic Aromatic Compound

Authors: Khaled S. Al Salhen

Abstract:

Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6µM) for hepatic aldehyde oxidase, will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: Aldehyde oxidase, Fish, Phenanthridine, Specificity.

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184 The Effects of Organic or Inorganic Zinc and Microbial Phytase, Alone or in Combination, on the Performance, Biochemical Parameters and Nutrient Utilization of Broilers Fed a Diet Low in Available Phosphorus

Authors: Mustafa Midilli, Mustafa Salman, Omer Hakan Muglali, Tülay Ögretmen, Sena Cenesiz, Neslihan Ormanci

Abstract:

This study examined the effects of zinc (Zn) from different sources and microbial phytase on the broiler performance, biochemical parameters and digestibility of nutrients when they were added to broiler diets containing low available phosphorus. A total of 875, 1-day-old male broilers of the Ross 308 strain were randomly separated into two control groups (positive and negative) and five treatment groups each containing 125 birds; each group was divided into 5 replicates of 25 birds. The positive control (PC) group was fed a diet containing adequate concentration (0.45%) of available phosphorus due to mineral premix (except zinc) and feeds. The negative control (NC) group was fed a basal diet including low concentration (0.30%) of available phosphorus due to mineral premix (except zinc) and feeds. The basal diet was supplemented with 0.30% phosphorus and 500 FTU phytase (PH); 0.30% phosphorus and organic zinc (OZ; 75mg/kg of Zn from Zn-proteinate); 0.30% phosphorus and inorganic zinc (IZ; 75 mg/kg of Zn from ZnSO4); 0.30% phosphorus, organic zinc and 500 FTU phytase (OZ + PH); and 0.30% phosphorus, inorganic zinc and 500 FTU phytase (IZ + PH) in the treatment groups 1, 2, 3, 4 and 5, respectively. The lowest value for mean body weight was in the negative control group on a diet containing low available phosphorus. The use of supplementation with organic and inorganic zinc alone or in combination with microbial phytase significantly (P<0.05) increased the digestibility of Zn in the male broilers. Supplementation of those diets with OZ + PH or IZ + PH was very effective for increasing the body weight, body weight gain and the feed conversion ratio. In conclusion, the effects on broilers of diets with low phosphorus levels may be overcome by the addition of inorganic or organic zinc compounds in combination with microbial phytase.

Keywords: Broiler, Performance, Phytase, Phosphorus, Zinc.

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183 In vitro Effects of Viscum album on the Functionality of Rabbit Spermatozoa

Authors: Marek Halenár, Eva Tvrdá, Simona Baldovská, Ľubomír Ondruška, Peter Massányi, Adriana Kolesárová

Abstract:

This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P<0.05 with respect to time 8h). At the same time, concentrations ranging between 1 and 100 µg/mL of the extract led to a significant preservation of the cell viability (P<0.05 in case of 5, 50 and 100 µg/mL; P<0.01 with respect to 1 and 10 µg/mL, time 8h). 1 and 5 µg/mL of the extract exhibited antioxidant characteristics, translated into a significant reduction of the ROS production, particularly notable at time 8h (P<0.01). The results indicate that the Viscum extract is capable of delaying the damage inflicted to the spermatozoon by the in vitro environment.

Keywords: CASA, mistletoe, mitochondrial activity, motility, reactive oxygen species, rabbits, spermatozoa, Viscum album.

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