Search results for: in vitro Rumen Fermentation
1557 Chemical and Sensory Properties of Chardonnay Wines Produced in Different Oak Barrels
Authors: Valentina Obradović, Josip Mesić, Maja Ergović Ravančić, Kamila Mijowska, Brankica Svitlica
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French oak and American oak barrels are most famous all over the world, but barrels of different origin can also be used for obtaining high quality wines. The aim of this research was to compare the influence of different Slavonian (Croatian) and French oak barrels on the quality of Chardonnay wine. Grapes were grown in Croatian wine growing region of Kutjevo in 2015. Chardonnay wines were tested for basic oenological parameters (alcohol, extract, reducing sugar, SO2, acidity), total polyphenols content (Folin-Ciocalteu method), antioxidant activity (ABTS and DPPH method) and color density. Sensory evaluation was performed by students of viticulture/oenology. Samples produced by classical fermentation and ageing in French oak barrels, had better results for polyphenols and sensory evaluation (especially low toasting level) than samples in Slavonian barrels. All tested samples were scored as a “quality” or “premium quality” wines. Sur lie method of fermentation and ageing in Slavonian oak barrel had very good extraction of polyphenols and high antioxidant activity with the usage of authentic yeasts, while commercial yeast strain resulted in worse chemical and sensory parameters.Keywords: chardonnay, French oak, Slavonian oak, sur lie
Procedia PDF Downloads 2421556 Beneficial Effect of Micropropagation Coupled with Mycorrhization on Enhancement of Growth Performance of Medicinal Plants
Authors: D. H. Tejavathi
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Medicinal plants are globally valuable sources of herbal products. Wild populations of many medicinal plants are facing threat of extinction because of their narrow distribution, endemicity, and degradation of specific habitats. Micropropagation is an established in vitro technique by which large number of clones can be obtained from a small bit of explants in a short span of time within a limited space. Mycorrhization can minimize the transient transplantation shock, experienced by the micropropagated plants when they are transferred from lab to land. AM fungal association improves the physiological status of the host plants through better uptake of water and nutrients, particularly phosphorus. Consequently, the growth performance and biosynthesis of active principles are significantly enhanced in AM fungal treated plants. Bacopa monnieri, Andrographis paniculata, Agave vera-curz, Drymaria cordata and Majorana hortensis, important medicinal plants used in various indigenous systems of medicines, are selected for the present study. They form the main constituents of many herbal formulations. Standard in vitro techniques were followed to obtain the micropropagated plants. Shoot tips and nodal segments were used as explants. Explants were cultured on Murashige and Skoog, and Phillips and Collins media supplemented with various combinations of growth regulators. Multiple shoots were obtained on a media containing both auxins and cytokinins at various concentrations and combinations. Multiple shoots were then transferred to rooting media containing auxins for root induction. Thus, obtained in vitro regenerated plants were subjected to brief acclimatization before transferring them to land. One-month-old in vitro plants were treated with AM fungi, and the symbiotic effect on the overall growth parameters was analyzed. It was found that micropropagation coupled with mycorrhization has significant effect on the enhancement of biomass and biosynthesis of active principles in these selected medicinal plants. In vitro techniques coupled with mycorrhization have opened a possibility of obtaining better clones in respect of enhancement of biomass and biosynthesis of active principles. Beneficial effects of AM fungal association with medicinal plants are discussed.Keywords: cultivation, medicinal plants, micropropagation, mycorrhization
Procedia PDF Downloads 1711555 Role of Fish Hepatic Aldehyde Oxidase in Oxidative In Vitro Metabolism of Phenanthridine Heterocyclic Aromatic Compound
Authors: Khaled S. Al Salhen
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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 ± 7.6 µM) for hepatic 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
Procedia PDF Downloads 3641554 Temporal Changes of Heterogeneous Subpopulations of Human Adipose-Derived Stromal/Stem Cells in vitro
Authors: Qiuyue Peng, Vladimir Zachar
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The application of adipose-derived stromal/stem cells (ASCs) in regenerative medicine is gaining more awareness due to their advanced translational potential and abundant source preparations. However, ASC-based translation has been confounded by high subpopulation heterogeneity, causing ambiguity about its precise therapeutic value. Some phenotypes defined by a unique combination of positive and negative surface markers have been found beneficial to the required roles. Therefore, the immunophenotypic repertoires of cultured ASCs and temporal changes of distinct subsets were investigated in this study. ASCs from three donors undergoing cosmetic liposuction were cultured in standard culturing methods, and the co-expression patterns based on the combination of selected markers at passages 1, 4, and 8 were analyzed by multi-chromatic flow cytometry. The results showed that the level of heterogeneity of subpopulations of ASCs became lower by in vitro expansion. After a few passages, most of the CD166⁺/CD274⁺/CD271⁺ based subpopulations converged to CD166 single positive cells. Meanwhile, these CD29⁺CD201⁺ double-positive cells, in combination with CD36/Stro-1 expression or without, feathered only the major epitopes and maintained prevailing throughout the whole process. This study suggested that, upon in vitro expansion, the phenotype repertoire of ASCs redistributed and stabilized in a way that cells co-expressing exclusively the strong markers remained dominant. These preliminary findings provide a general overview of the distribution of heterogeneous subsets residents within human ASCs during expansion in vitro. It is a critical step to fully characterize ASCs before clinical application, although the biological effects of heterogeneous subpopulations still need to be clarified.Keywords: adipose-derived stromal/stem cells, heterogeneity, immunophenotype, subpopulations
Procedia PDF Downloads 1141553 Effects of Bacterial Inoculants and Enzymes Inoculation on the Fermentation and Aerobic Stability of Potato Hash Silage
Authors: B. D. Nkosi, T. F. Mutavhatsindi, J. J. Baloyi, R. Meeske, T. M. Langa, I. M. M. Malebana, M. D. Motiang
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Potato hash (PH), a by-product from food production industry, contains 188.4 g dry matter (DM)/kg and 3.4 g water soluble carbohydrate (WSC)/kg DM, and was mixed with wheat bran (70:30 as is basis) to provide 352 g DM/kg and 315 g WSC/kg DM. The materials were ensiled with or without silage additives in 1.5L anaerobic jars (3 jars/treatment) that were kept at 25-280 C for 3 months. Four types of silages were produced which were: control (no additive, denoted as T1), celluclast enzyme (denoted as T2), emsilage bacterial inoculant (denoted as T3) and silosolve bacterial inoculant (denoted as T4). Three jars per treatment were opened after 3 months of ensiling for the determination of nutritive values, fermentation characteristics and aerobic stability. Aerobic stability was done by exposing silage samples to air for 5 days. The addition of enzyme (T2) was reduced (P<0.05) silage pH, fiber fractions (NDF and ADF) while increasing (P < 0.05) residual WSC and lactic acid (LA) production, compared to other treatments. Silage produced had pH of < 4.0, indications of well-preserved silage. Bacterial inoculation (T3 and T4) improved (P < 0.05) aerobic stability of the silage, as indicated by increased number of hours and lower CO2 production, compared to other treatments. However, the aerobic stability of silage was worsen (P < 0.05) with the addition of an enzyme (T2). Further work to elucidate these effects on nutrient digestion and growth performance on ruminants fed the silage is needed.Keywords: by-products, digestibility, feeds, inoculation, ruminants, silage
Procedia PDF Downloads 4391552 Biotransformation of Glycerine Pitch as Renewable Carbon Resource into P(3HB-co-4HB) Biopolymer
Authors: Amirul Al-Ashraf Abdullah, Hema Ramachandran, Iszatty Ismail
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Oleochemical industry in Malaysia has been diversifying significantly due to the abundant supply of both palm and kernel oils as raw materials as well as the high demand for downstream products such as fatty acids, fatty alcohols and glycerine. However, environmental awareness is growing rapidly in Malaysia because oleochemical industry is one of the palm-oil based industries that possess risk to the environment. Glycerine pitch is one of the scheduled wastes generated from the fatty acid plants in Malaysia and its discharge may cause a serious environmental problem. Therefore, it is imperative to find alternative applications for this waste glycerine. Consequently, the aim of this research is to explore the application of glycerine pitch as direct fermentation substrate in the biosynthesis of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer, aiming to contribute toward the sustainable production of biopolymer in the world. Utilization of glycerine pitch (10 g/l) together with 1,4-butanediol (5 g/l) had resulted in the achievement of 40 mol% 4HB monomer with the highest PHA concentration of 2.91 g/l. Synthesis of yellow pigment which exhibited antimicrobial properties occurred simultaneously with the production of P(3HB-co-4HB) through the use of glycerine pitch as renewable carbon resource. Utilization of glycerine pitch in the biosynthesis of P(3HB-co-4HB) will not only contribute to reducing society’s dependence on non-renewable resources but also will promote the development of cost efficiency microbial fermentation towards biosustainability and green technology.Keywords: biopolymer, glycerine pitch, natural pigment, P(3HB-co-4HB)
Procedia PDF Downloads 4691551 Morphological Interaction of Porcine Oocyte and Cumulus Cells Study on in vitro Oocyte Maturation Using Electron Microscopy
Authors: M. Areekijseree, W. Pongsawat, M. Pumipaiboon, C. Thepsithar, S. Sengsai, T. Chuen-Im
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Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by zona pellucida with layer of cumulus cells ranging between 59.29-202.14 µm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 µg/mL porcine follicle-stimulating hormone, 1 µg/mL LH, 1µg/mL estradiol with ethanol, and 50 µg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.Keywords: cumulus cells, electron microscopy, in vitro, porcine oocyte
Procedia PDF Downloads 3851550 Development and in vitro Characterization of Loteprednol Etabonate-Loaded Polymeric Nanoparticles for Ocular Delivery
Authors: Abhishek Kumar Sah, Preeti K. Suresh
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Effective drug delivery to the eye is a massive challenge, due to complicated physiological ocular barriers, rapid washout by tear and nasolachrymal drainage. Thus, most of the conventional ophthalmic formulations face the problem of low ocular bioavailability. Ophthalmic drug therapy can be improved by enhancing the precorneal drug retention along with improved drug penetration. The aim of the present investigation was to develop and evaluate a biodegradable polymer poly (D, L-lactide-co-glycolide) (PLGA) coated nanoparticulate carrier of loteprednol etabonate. PLGA nanoparticles were prepared by modified emulsification/solvent diffusion method using high-speed homogenizer followed by sonication. The nanoparticles were characterized for various parameters such as particle size, zeta potential, polydispersity index, X-ray powder diffraction (XRD), Transmission electron microscopy (TEM), in vitro drug release profile and stability. The prepared nanocarriers displayed mean particle size in the range of 271.7 to 424.4 nm, with zeta potential less than –10 mV. In vitro release in simulated tear fluid (STF) nanocarrier showed an extended release profile of loteprednol etabonate. TEM confirmed the spherical morphology and smooth surface of the particles. All the prepared formulations were found to be stable at varying temperatures.Keywords: drug delivery, ocular delivery, polymeric nanoparticles, loteprednol etabonate
Procedia PDF Downloads 5511549 Inhibitory Impacts of Fulvic Acid-Coated Iron Oxide Nano Particles on the Amyloid Fibril Aggregations
Authors: Dalia Jomehpour, Sara Sheikhlary, Esmaeil Heydari, Mohammad Hossien Majles Ara
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In this study, we report fulvic acid-coated iron oxide nanoparticles of 10.7 ± 2.7 nm size, which serve to inhibit amyloid fibrillation formation. Although the effect of fulvic acid on tau fibrils was investigated, to our best knowledge, its inhibitory impacts on amyloid aggregation formation have been assessed neither in-vitro nor in-vivo. On the other hand, iron oxide nanoparticles exhibit anti-amyloid activity on their own. This study investigates the inhibitory effect of fulvic acid coated iron oxide nanoparticles on amyloid aggregations formed from the commonly used in-vitro model, lysozyme from chicken egg white. FESEM, XRD, and FTIR characterization confirmed that fulvic acid was coated onto the surface of the nanoparticles. The inhibitory effects of the fulvic acid coated iron oxide nanoparticles were verified by Thioflavin T assay, circular dichroism (CD), and FESEM analysis. Furthermore, the toxicity of the nanoparticles on the neuroblastoma SH-SY5Y human cell line was assessed through an MTT assay. Our results indicate that fulvic acid coated iron oxide nanoparticles can efficiently inhibit the formation of amyloid aggregations while exhibiting negligible in-vitro toxicity; thus, they can be used as anti-amyloid agents in the development of the potential drug for neurodegenerative diseases.Keywords: Alzheimer’s disease, fulvic acid coated iron oxide nanoparticles, fulvic acid, amyloid inhibitor, polyphenols
Procedia PDF Downloads 1121548 An Efficient and Low Cost Protocol for Rapid and Mass in vitro Propagation of Hyssopus officinalis L.
Authors: Ira V. Stancheva, Ely G. Zayova, Maria P. Geneva, Marieta G. Hristozkova, Lyudmila I. Dimitrova, Maria I. Petrova
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The study describes a highly efficient and low-cost protocol for rapid and mass in vitro propagation of medicinal and aromatic plant species (Hyssopus officinalis L., Lamiaceae). Hyssop is an important aromatic herb used for its medicinal values because of its antioxidant, anti-inflammatory and antimicrobial properties. The protocol for large-scale multiplication of this aromatic plant was developed using young stem tips explants. The explants were sterilized with 0.04% mercuric chloride (HgCl₂) solution for 20 minutes and washing three times with sterile distilled water in 15 minutes. The cultural media was full and half strength Murashige and Skoog medium containing indole-3-butyric acid. Full and ½ Murashige and Skoog media without auxin were used as controls. For each variant 20 glass tubes with two plants were used. In each tube two tip and nodal explants were inoculated. Maximum shoot and root number were obtained on ½ Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid at the same time after four weeks of culture. The number of shoots per explant and shoot height were considered. The data on rooting percentage, the number of roots per plant and root length were collected after the same cultural period. The highest percentage of survival 85% for this medicinal plant was recorded in mixture of soil, sand and perlite (2:1:1 v/v/v). This mixture was most suitable for acclimatization of all propagated plants. Ex vitro acclimatization was carried out at 24±1 °C and 70% relative humidity under 16 h illuminations (50 μmol m⁻²s⁻¹). After adaptation period, the all plants were transferred to the field. The plants flowered within three months after transplantation. Phenotypic variations in the acclimatized plants were not observed. An average of 90% of the acclimatized plants survived after transferring into the field. All the in vitro propagated plants displayed normal development under the field conditions. Developed in vitro techniques could provide a promising alternative tool for large-scale propagation that increases the number of homologous plants for field cultivation. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.Keywords: Hyssopus officinalis L., in vitro culture, micro propagation, acclimatization
Procedia PDF Downloads 3121547 PLA Production from Multi Supply Lignocellulosic Biomass Residues: A Pathway for Agrifood Sector
Authors: Sónia Ribeiro, Diana Farinha, Hélia Sales, Rita Pontes, João Nunes
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The demand and commitment to sustainability in the agrifood sector introduce news opportunities for new composite materials. Composite materials are emerging as a vital entity for the sustainable development. Polylactic acid (PLA) has been recognized as a potential polymer with attractive characteristics for agrifood sector applications. PLA that can be beneficial for the development of composites, biocomposites, films, porous gels, and so on. The production of PLA from lignocellulosic biomass residues matrix is a key option towards a sustainable and circular bioeconomy and a non-competitive application with feed and food sector. The Flui and BeirInov projects presents news developments in the production of PLA composites to value the Portuguese forest ecosystem, with high amount of lignocellulosic biomass residues and available. A performance production of lactic acid from lignocellulosic biomass undergoes a process of autohydrolysis, saccharification and fermentation, originating a lactic acid fermentation medium with a 72.27g.L-1 was obtained and a final purification of 72%. The high purification PLA from multi lignocellulosic residues representing one economic expensive process, and a new materials and application for the polymers and a combination with others types of composites matrix characteristic is the drive-up for this green market.Keywords: polylactic acid, lignocellulosic biomass, agrifood, composite materials
Procedia PDF Downloads 751546 Effects of in silico (Virtual Lab) And in vitro (inside the Classroom) Labs in the Academic Performance of Senior High School Students in General Biology
Authors: Mark Archei O. Javier
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The Fourth Industrial Revolution (FIR) is a major industrial era characterized by the fusion of technologies that is blurring the lines between the physical, digital, and biological spheres. Since this era teaches us how to thrive in the fast-paced developing world, it is important to be able to adapt. With this, there is a need to make learning and teaching in the bioscience laboratory more challenging and engaging. The goal of the research is to find out if using in silico and in vitro laboratory activities compared to the conventional conduct laboratory activities would have positive impacts on the academic performance of the learners. The potential contribution of the research is that it would improve the teachers’ methods in delivering the content to the students when it comes to topics that need laboratory activities. This study will develop a method by which teachers can provide learning materials to the students. A one-tailed t-Test for independent samples was used to determine the significant difference in the pre- and post-test scores of students. The tests of hypotheses were done at a 0.05 level of significance. Based on the results of the study, the gain scores of the experimental group are greater than the gain scores of the control group. This implies that using in silico and in vitro labs for the experimental group is more effective than the conventional method of doing laboratory activities.Keywords: academic performance, general biology, in silico laboratory, in vivo laboratory, virtual laboratory
Procedia PDF Downloads 1891545 Combining in vitro Protein Expression with AlphaLISA Technology to Study Protein-Protein Interaction
Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov
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The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction
Procedia PDF Downloads 2371544 Effect of Different Processing Methods on the Proximate, Functional, Sensory, and Nutritional Properties of Weaning Foods Formulated from Maize (Zea mays) and Soybean (Glycine max) Flour Blends
Authors: C. O. Agu, C. C. Okafor
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Maize and soybean flours were produced using different methods of processing which include fermentation (FWF), roasting (RWF) and malting (MWF). Products from the different methods were mixed in the ratio 60:40 maize/soybean, respectively. These composites mixed with other ingredients such as sugar, vegetable oil, vanilla flavour and vitamin mix were analyzed for proximate composition, physical/functional, sensory and nutritional properties. The results for the protein content ranged between 6.25% and 16.65% with sample RWF having the highest value. Crude fibre values ranged from 3.72 to 10.0%, carbohydrate from 58.98% to 64.2%, ash from 1.27 to 2.45%. Physical and functional properties such as bulk density, wettability, gelation capacity have values between 0.74 and 0.76g/ml, 20.33 and 46.33 min and 0.73 to 0.93g/ml, respectively. On the sensory quality colour, flavour, taste, texture and general acceptability were determined. In terms of colour and flavour there was no significant difference (P < 0.05) while the values for taste ranged between 4.89 and 7.1 l, texture 5.50 to 8.38 and general acceptability 6.09 and 7.89. Nutritionally there is no significant difference (P < 0.05) between sample RWF and the control in all parameters considered. Samples FWF and MWF showed significantly (P < 0.5) lower values in all parameters determined. In the light of the above findings, roasting method is highly recommend in the production of weaning foods.Keywords: fermentation, malting, ratio, roasting, wettability
Procedia PDF Downloads 3041543 Evaluation of Naringenin Role in Inhibiton of Lung Tumor Progression in Mice
Authors: Vishnu Varthan Vaithiyalingamjagannathan, M. N. Sathishkumar, K. S. Lakhsmi, D. Satheeshkumar, Srividyaammayappanrajam
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Background:Naringenin, aglycone flavonoid possess certain activities like anti-oxidant, anti-estrogenic, anti-diabetic, cardioprotective, anti-obesity,anti-inflammatory, hepatoprotective and also have anti-cancer characteristics like carcinogenic inactivation, cell cycle arrest, anti-proliferation, apoptosis, anti-angiogenesis and enhances anti-oxidant activity. Methodology:The inhibitory effect of Naringenin in lung tumor progression estimated with adenocarcinoma (A549) cell lines (in vitro) and C57BL/6 mice injected with 5 X 106A549 cell lines (in vivo) in a tri-dose manner (Naringenin 100mg/kg,150mg/kg, and 200mg/kg) compared with standard chemotherapy drug cisplatin (7mg/kg). Results:The results of the present study revealed a dose-dependent activity in Naringenin and combination with cisplatin at a higher dose which showed decreased tumor progression in mice. In vitro studies carried out for estimation of cell survival and Nitric Oxide (NO) level, shows dose dependent action of Naringenin with IC50 value of 42µg/ml. In vivo studies were carried out in C57BL/6 mice. Naringenin satisfied the condition of an anti-cancer molecule with its characteristics in fragmentation assay, Zymography assay, anti-oxidant, and myeloperoxidase studies, than cisplatin which failed in anti-oxidant and myeloperoxidase effect. Both in vitro and in vivo establishes dose dependent decrease in NO levels. But whereas, Naringenin showed adverse results in Matrix Metalloproteinase (MMP) enzymatic levels with increase in dose levels. Conclusion:From the present study, Naringenin could suppress the lung tumor progression when given individually and also in combinatorial with standard chemotherapy drug.Keywords: naringenin, in vitro, cell line, anticancer
Procedia PDF Downloads 4351542 The Effects of in vitro Digestion on Cheese Bioactivity; Comparing Adult and Elderly Simulated in vitro Gastrointestinal Digestion Models
Authors: A. M. Plante, F. O’Halloran, A. L. McCarthy
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By 2050 it is projected that 2 billion of the global population will be more than 60 years old. Older adults have unique dietary requirements and aging is associated with physiological changes that affect appetite, sensory perception, metabolism, and digestion. Therefore, it is essential that foods recommended and designed for older adults promote healthy aging. To assess cheese as a functional food for the elderly, a range of commercial cheese products were selected and compared for their antioxidant properties. Cheese from various milk sources (bovine, goats, sheep) with different textures and fat content, including cheddar, feta, goats, brie, roquefort, halloumi, wensleydale and gouda, were initially digested with two different simulated in vitro gastrointestinal digestion (SGID) models. One SGID model represented a validated in vitro adult digestion system and the second model, an elderly SGID, was designed to consider the physiological changes associated with aging. The antioxidant potential of all cheese digestates was investigated using in vitro chemical-based antioxidant assays, (2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) and total phenolic content (TPC)). All adult model digestates had high antioxidant activity across both DPPH ( > 70%) and FRAP ( > 700 µM Fe²⁺/kg.fw) assays. Following in vitro digestion using the elderly SGID model, full-fat red cheddar, low-fat white cheddar, roquefort, halloumi, wensleydale, and gouda digestates had significantly lower (p ≤ 0.05) DPPH radical scavenging properties compared to the adult model digestates. Full-fat white cheddar had higher DPPH radical scavenging activity following elderly SGID digestion compared to the adult model digestate, but the difference was not significant. All other cheese digestates from the elderly model were comparable to the digestates from the adult model in terms of radical scavenging activity. The FRAP of all elderly digestates were significantly lower (p ≤ 0.05) compared to the adult digestates. Goats cheese was significantly higher (p ≤ 0.05) in FRAP (718 µM Fe²/kg.fw) compared to all other digestates in the elderly model. TPC levels in the soft cheeses (feta, goats) and low-fat cheeses (red cheddar, white cheddar) were significantly lower (p ≤ 0.05) in the elderly digestates compared to the adult digestates. There was no significant difference in TPC levels, between the elderly and adult model for full-fat cheddar (red, white), roquefort, wensleydale, gouda, and brie digestates. Halloumi cheese was the only cheese that was significantly higher in TPC levels following elderly digestion compared to adult digestates. Low fat red cheddar had significantly higher (p ≤ 0.05) TPC levels compared to all other digestates for both adult and elderly digestive systems. Findings from this study demonstrate that aging has an impact on the bioactivity of cheese, as antioxidant activity and TPC levels were lower, following in vitro elderly digestion compared to the adult model. For older adults, soft cheese, particularly goats cheese, was associated with high radical scavenging and reducing power, while roquefort cheese had low antioxidant activity. Also, elderly digestates of halloumi and low-fat red cheddar were associated with high TPC levels. Cheese has potential as a functional food for the elderly, however, bioactivity can vary depending on the cheese matrix. Funding for this research was provided by the RISAM Scholarship Scheme, Cork Institute of Technology, Ireland.Keywords: antioxidants, cheese, in-vitro digestion, older adults
Procedia PDF Downloads 2281541 Fermented Fruit and Vegetable Discard as a Source of Feeding Ingredients and Functional Additives
Authors: Jone Ibarruri, Mikel Manso, Marta Cebrián
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A high amount of food is lost or discarded in the World every year. In addition, in the last decades, an increasing demand of new alternative and sustainable sources of proteins and other valuable compounds is being observed in the food and feeding sectors and, therefore, the use of food by-products as nutrients for these purposes sounds very interesting from the environmental and economical point of view. However, the direct use of discarded fruit and vegetables that present, in general, a low protein content is not interesting as feeding ingredient except if they are used as a source of fiber for ruminants. Especially in the case of aquaculture, several alternatives to the use of fish meal and other vegetable protein sources have been extensively explored due to the scarcity of fish stocks and the unsustainability of fishing for these purposes. Fish mortality is also of great concern in this sector as this problem highly reduces their economic feasibility. So, the development of new functional and natural ingredients that could reduce the need for vaccination is also of great interest. In this work, several fermentation tests were developed at lab scale using a selected mixture of fruit and vegetable discards from a wholesale market located in the Basque Country to increase their protein content and also to produce some bioactive extracts that could be used as additives in aquaculture. Fruit and vegetable mixtures (60/40 ww) were centrifugated for humidity reduction and crushed to 2-5 mm particle size. Samples were inoculated with a selected Rhizopus oryzae strain and fermented for 7 days in controlled conditions (humidity between 65 and 75% and 28ºC) in Petri plates (120 mm) by triplicate. Obtained results indicated that the final fermented product presented a twofold protein content (from 13 to 28% d.w). Fermented product was further processed to determine their possible functionality as a feed additive. Extraction tests were carried out to obtain an ethanolic extract (60:40 ethanol: water, v.v) and remaining biomass that also could present applications in food or feed sectors. The extract presented a polyphenol content of about 27 mg GAE/gr d.w with antioxidant activity of 8.4 mg TEAC/g d.w. Remining biomass is mainly composed of fiber (51%), protein (24%) and fat (10%). Extracts also presented antibacterial activity according to the results obtained in Agar Diffusion and to the Minimum Inhibitory Concentration (MIC) tests determined against several food and fish pathogen strains. In vitro, digestibility was also assessed to obtain preliminary information about the expected effect of extraction procedure on fermented product digestibility. First results indicated that remaining biomass after extraction doesn´t seem to improve digestibility in comparison to the initial fermented product. These preliminary results show that fermented fruit and vegetables can be a useful source of functional ingredients for aquaculture applications and a substitute of other protein sources in the feeding sector. Further validation will be also carried out through “in vivo” tests with trout and bass.Keywords: fungal solid state fermentation, protein increase, functional extracts, feed ingredients
Procedia PDF Downloads 641540 Development of in vitro Fertilization and Emerging Legal Issues
Authors: Malik Imtiaz Ahmad
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The development of In Vitro Fertilization (IVF) has revolutionized the field of reproductive medicine, offering hope to myriad individuals and couples facing infertility issues. IVF, a process involving the fertilization of eggs with sperm outside the body, has evolved over decades from an experimental procedure to a mainstream medical practice. The study sought to understand the evolution of IVF from its early stages to its present status as a groundbreaking fertility treatment. It also aimed to analyze the legal complexities surrounding IVF, including issues like embryo ownership, surrogacy agreements, and custody disputes. This research focused on the multidisciplinary approach involving both medical and legal fields. It aimed to explore the historical evolution of IVF, its techniques, and legal challenges concerning family law, health law, and privacy policies it has given rise to in modern times. This research aimed to provide insights into the intersection of medical technology and the law, offering valuable knowledge for policymakers, legal experts, and individuals involved in IVF. The study utilized various methods, including a thorough literature review, a historical analysis of IVF’s evolution, an examination of legal cases, and a review of emerging regulations. These approaches aimed to provide a comprehensive understanding of IVF and its modern legal issues, facilitating a holistic exploration of the subject matter.Keywords: in vitro fertilization development, IVF techniques evolution, legal issues in IVF, IVF legal frameworks, ethical dilemmas in IVF
Procedia PDF Downloads 351539 Bismuth-Inhibitory Effects on Bacteria and Stimulation of Fungal Growth In vitro
Authors: Sulaiman B. Ali Alharbi, Bassam H. Mashat, Naif Abdullah Al-Harbi, Milton Wainwright, Abeer S. Aloufi, Sulamain Alnaimat
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Bismuth salicylate was found to inhibit the growth of a range of bacteria and yeast, Candida albican. In general the growth of bacteria did not result in the increase in bismuth solubilisation, in contrast, bismuth solubilisation increased following the growth of C. albicans. A significant increase in the biomass (dry weight) of Aspergillus niger and Aspergillus oryzae occurred in vitro when these fungi were grown in the presence of bismuth salicylate. Biomass increase occurred over a range of bismuth compound additions, which in the case of A. oryzae was associated with the increase in the solubilisation of the insoluble bismuth compounds.Keywords: bacterial inhibition, fungal growth stimulation, medical uses of bismuth, yeast inhibition
Procedia PDF Downloads 3411538 Effects of Different Food Matrices on Viscosity and Protein Degradation during in vitro Digestion
Authors: Gulay Oncu Ince, Sibel Karakaya
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Food is a worldwide concern. Among the factors that have influences on human health, food, nutrition and life style have been regarded as the most important factors since they can be intervened. While some parts of the world has been faced with food shortages and hence, chronic metabolic diseases, the other part of the world have been emerged from over consumption of food. Both situations can result in shorter life expectancy and represent a major global health problem. Hunger, satiety and appetite sensation form a balance ensures the operation of feeding behavior between food intake and energy consumption. Satiety is one of the approaches that is effective in ensuring weight control and avoid eating more in the postprandial period. By manipulating the microstructure of food macro and micronutrient bioavailability may be increased or reduced. For the food industry appearance, texture, taste structural properties as well as the gastrointestinal tract behavior of the food after the consumption is becoming increasingly important. Also, this behavior has been the subject of several researches in recent years by the scientific community. Numerous studies have been published about changing the food matrix in order to increase expected impacts. In this study, yogurts were enriched with caseinomacropeptide (CMP), whey protein (WP), CMP and sodium alginate (SA), and WP + SA in order to produce goat yogurts having different food matrices. SDS Page profiles of the samples after in vitro digestion and viscosities of the stomach digesta at different share rates were determined. Energy values were 62.11kcal/100 g, 70.27 kcal/100 g, 70.61 kcal/100 g, 71.20 kcal/100 g and 71.67 kcal/100 g for control, CMP added WP added, WP + SA added, and CMP + SA added yogurts respectively. The results of viscosity analysis showed that control yogurt had the lowest viscosity value and this was followed by CMP added, WP added, CMP + SA added and WP + SA added yogurts, respectively. Protein contents of the stomach and duedonal digests of the samples after subjected to two different in vitro digestion methods were changed between 5.34-5.91 mg protein / g sample and 16.93-19.75 mg protein /g of sample, respectively. Viscosity measurements of the stomach digests showed that CMP + SA added yogurt displayed the highest viscosity value in both in vitro digestion methods. There were differences between the protein profiles of the stomach and duedonal digests obtained by two different in vitro digestion methods (p<0.05).Keywords: caseinomacropeptide, protein profile, whey protein, yogurt
Procedia PDF Downloads 4891537 Effect of Mannitol on in Vitro Conservation of Local and Exotic Taro-Genotypes (Colocasia Esculenta Var Esculenta)
Authors: Benjamin Bonsu Bruce, Marian Dorcas Quain David Appiah-Kubi, Gertrude Osei-Diko, Harrison Kwame Dapaah
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Taro [Colocasia esculenta (L.) Schott] is a major staple food and remains a significant crop to many cultural and agricultural customs worldwide. In Ghana, taro is mostly propagated using vegetative material, which is conserved in field collection and recycled from their farms to establish new fields. However, this practice promotes the accumulation of systemic pathogens. Prior exposure to pests and subsequent expression of disease symptoms can also be a huge constraint to sustainable conservation and utilization of taro genetic resources. In vitro, slow growth is one of the most promising techniques to be utilized for conservation. The objective of this study was to find a medium-term in vitro conservation protocol for local and exotic taro genotypes. The medium-term conservation study was conducted using actively growing shoots obtained from in vitro cultures. Explants were cultured to full strength in complete Murashige and Skoog medium supplemented with Mannitol at different concentrations (0g/l, 20g/l, 25g/l, and 30g/l). Another medium that was tested as an additional treatment is the White’s medium. The highest number of shoots (6.33) and leaves (22.67) occurred on medium containing 20 and 25g/l mannitol in genotype SAO 006 as compared to other genotypes, whereas 30g/l mannitol was the best to restrict growth for the entire 6 months period in terms of shoot height (22.50cm). The study reveals that mannitol supplemented culture media could reduce the growth of Colocasia plantlets, especially in stem height. Culture growth following 6 months of conservation, showed that healthy shoot cultures of Taro were obtained after 6 months of storage in a medium containing 20gl⁻¹ and 25gl⁻¹ mannitol.Keywords: complete murashige, skoog medium, culture conditions, mannitol, slow growth conservation
Procedia PDF Downloads 1701536 Reactors with Effective Mixing as a Solutions for Micro-Biogas Plant
Authors: M. Zielinski, M. Debowski, P. Rusanowska, A. Glowacka-Gil, M. Zielinska, A. Cydzik-Kwiatkowska, J. Kazimierowicz
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Technologies for the micro-biogas plant with heating and mixing systems are presented as a part of the Research Coordination for a Low-Cost Biomethane Production at Small and Medium Scale Applications (Record Biomap). The main objective of the Record Biomap project is to build a network of operators and scientific institutions interested in cooperation and the development of promising technologies in the sector of small and medium-sized biogas plants. The activities carried out in the project will bridge the gap between research and market and reduce the time of implementation of new, efficient technological and technical solutions. Reactor with simultaneously mixing and heating system is a concrete tank with a rectangular cross-section. In the reactor, heating is integrated with the mixing of substrate and anaerobic sludge. This reactor is solution dedicated for substrates with high solids content, which cannot be introduced to the reactor with pumps, even with positive displacement pumps. Substrates are poured to the reactor and then with a screw pump, they are mixed with anaerobic sludge. The pumped sludge, flowing through the screw pump, is simultaneously heated by a heat exchanger. The level of the fermentation sludge inside the reactor chamber is above the bottom edge of the cover. Cover of the reactor is equipped with the screw pump driver. Inside the reactor, an electric motor is installed that is driving a screw pump. The heated sludge circulates in the digester. The post-fermented sludge is collected using a drain well. The inlet to the drain well is below the level of the sludge in the digester. The biogas is discharged from the reactor by the biogas intake valve located on the cover. The technology is very useful for fermentation of lignocellulosic biomass and substrates with high content of dry mass (organic wastes). The other technology is a reactor for micro-biogas plant with a pressure mixing system. The reactor has a form of plastic or concrete tank with a circular cross-section. The effective mixing of sludge is ensured by profiled at 90° bottom of the tank. Substrates for fermentation are supplied by an inlet well. The inlet well is equipped with a cover that eliminates odour release. The introduction of a new portion of substrates is preceded by pumping of digestate to the disposal well. Optionally, digestate can gravitationally flow to digestate storage tank. The obtained biogas is discharged into the separator. The valve supplies biogas to the blower. The blower presses the biogas from the fermentation chamber in such a way as to facilitate the introduction of a new portion of substrates. Biogas is discharged from the reactor by valve that enables biogas removal but prevents suction from outside the reactor.Keywords: biogas, digestion, heating system, mixing system
Procedia PDF Downloads 1541535 Cybernetic Modeling of Growth Dynamics of Debaryomyces nepalensis NCYC 3413 and Xylitol Production in Batch Reactor
Authors: J. Sharon Mano Pappu, Sathyanarayana N. Gummadi
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Growth of Debaryomyces nepalensis on mixed substrates in batch culture follows diauxic pattern of completely utilizing glucose during the first exponential growth phase, followed by an intermediate lag phase and a second exponential growth phase consuming xylose. The present study deals with the development of cybernetic mathematical model for prediction of xylitol production and yield. Production of xylitol from xylose in batch fermentation is investigated in the presence of glucose as the co-substrate. Different ratios of glucose and xylose concentrations are assessed to study the impact of multi substrate on production of xylitol in batch reactors. The parameters in the model equations were estimated from experimental observations using integral method. The model equations were solved simultaneously by numerical technique using MATLAB. The developed cybernetic model of xylose fermentation in the presence of a co-substrate can provide answers about how the ratio of glucose to xylose influences the yield and rate of production of xylitol. This model is expected to accurately predict the growth of microorganism on mixed substrate, duration of intermediate lag phase, consumption of substrate, production of xylitol. The model developed based on cybernetic modelling framework can be helpful to simulate the dynamic competition between the metabolic pathways.Keywords: co-substrate, cybernetic model, diauxic growth, xylose, xylitol
Procedia PDF Downloads 3281534 Co-Synthesis of Exopolysaccharides and Polyhydroxyalkanoates Using Waste Streams: Solid-State Fermentation as an Alternative Approach
Authors: Laura Mejias, Sandra Monteagudo, Oscar Martinez-Avila, Sergio Ponsa
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Bioplastics are gaining attention as potential substitutes of conventional fossil-derived plastics and new components of specialized applications in different industries. Besides, these constitute a sustainable alternative since they are biodegradable and can be obtained starting from renewable sources. Thus, agro-industrial wastes appear as potential substrates for bioplastics production using microorganisms, considering they are a suitable source for nutrients, low-cost, and available worldwide. Therefore, this approach contributes to the biorefinery and circular economy paradigm. The present study assesses the solid-state fermentation (SSF) technology for the co-synthesis of exopolysaccharides (EPS) and polyhydroxyalkanoates (PHA), two attractive biodegradable bioplastics, using the leftover of the brewery industry brewer's spent grain (BSG). After an initial screening of diverse PHA-producer bacteria, it was found that Burkholderia cepacia presented the highest EPS and PHA production potential via SSF of BSG. Thus, B. cepacia served to identify the most relevant aspects affecting the EPS+PHA co-synthesis at a lab-scale (100g). Since these are growth-dependent processes, they were monitored online through oxygen consumption using a dynamic respirometric system, but also quantifying the biomass production (gravimetric) and the obtained products (EtOH precipitation for EPS and solid-liquid extraction coupled with GC-FID for PHA). Results showed that B. cepacia has grown up to 81 mg per gram of dry BSG (gDM) at 30°C after 96 h, representing up to 618 times higher than the other tested strains' findings. Hence, the crude EPS production was 53 mg g-1DM (2% carbohydrates), but purity reached 98% after a dialysis purification step. Simultaneously, B. cepacia accumulated up to 36% (dry basis) of the produced biomass as PHA, mainly composed of polyhydroxybutyrate (P3HB). The maximum PHA production was reached after 48 h with 12.1 mg g⁻¹DM, representing threefold the levels previously reported using SSF. Moisture content and aeration strategy resulted in the most significant variables affecting the simultaneous production. Results show the potential of co-synthesis via SSF as an attractive alternative to enhance bioprocess feasibility for obtaining these bioplastics in residue-based systems.Keywords: bioplastics, brewer’s spent grain, circular economy, solid-state fermentation, waste to product
Procedia PDF Downloads 1441533 The Interplay of Dietary Fibers and Intestinal Microbiota Affects Type 2 Diabetes by Generating Short-Chain Fatty Acids
Authors: Muhammad Mazhar, Yong Zhu, Likang Qin
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Foods contain endogenous components known as dietary fibers, which are classified into soluble and insoluble forms. Dietary fibers are resistant to gut digestive enzymes, modulating anaerobic intestinal microbiota (AIM) and fabricating short-chain fatty acids (SCFAs). Acetate, butyrate, and propionate dominate in the gut, and different pathways, including Wood-Ljungdahl and acrylate pathways, generate these SCFAs. In pancreatic dysfunction, the release of insulin/glucagon is impaired, which leads to hyperglycemia. SCFAs enhance insulin sensitivity or secretion, beta-cell functions, leptin release, mitochondrial functions, and intestinal gluconeogenesis in human organs, which positively affect type 2 diabetes (T2D). Research models presented that SCFAs either enhance the release of peptide YY (PYY) and glucagon-like peptide-1 (GLP-1) from L-cells (entero-endocrine) or promote the release of leptin hormone satiation in adipose tissues through G-protein receptors, i.e., GPR-41/GPR-43. Dietary fibers are the components of foods that influence AIM and produce SCFAs, which may be offering beneficial effects on T2D. This review addresses the effectiveness of SCFAs in modulating gut AIM in the fermentation of dietary fiber and their worth against T2D.Keywords: dietary fibers, intestinal microbiota, short-chain fatty acids, fermentation, type 2 diabetes
Procedia PDF Downloads 731532 Genetically Modified Fuel-Ethanol Industrial Yeast Strains as Biocontrol Agents
Authors: Patrícia Branco, Catarina Prista, Helena Albergaria
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Industrial fuel-ethanol fermentations are carried out under non-sterile conditions, which favors the development of microbial contaminants, leading to huge economic losses. Wild yeasts such as Brettanomyces bruxellensis and lactic acid bacteria are the main contaminants of industrial bioethanol fermentation, affecting Saccharomyces cerevisiae performance and decreasing ethanol yields and productivity. In order to control microbial contaminations, the fuel-ethanol industry uses different treatments, including acid washing and antibiotics. However, these control measures carry environmental risks such as acid toxicity and the rise of antibiotic-resistant bacteria. Therefore, it is crucial to develop and apply less toxic and more environmentally friendly biocontrol methods. In the present study, an industrial fuel-ethanol starter, S. cerevisiae Ethanol-Red, was genetically modified to over-express AMPs with activity against fuel-ethanol microbial contaminants and evaluated regarding its biocontrol effect during mixed-culture alcoholic fermentations artificially contaminated with B. bruxellensis. To achieve this goal, S. cerevisiae Ethanol-Red strain was transformed with a plasmid containing the AMPs-codifying genes, i.e., partial sequences of TDH1 (925-963 bp) and TDH2/3 (925-963 bp) and a geneticin resistance marker. The biocontrol effect of those genetically modified strains was evaluated against B. bruxellensis and compared with the antagonistic effect exerted by the modified strain with an empty plasmid (without the AMPs-codifying genes) and the non-modified strain S. cerevisiae Ethanol-Red. For that purpose, mixed-culture alcoholic fermentations were performed in a synthetic must use the modified S. cerevisiae Ethanol-Red strains together with B. bruxellensis. Single-culture fermentations of B. bruxellensis strains were also performed as a negative control of the antagonistic effect exerted by S. cerevisiae strains. Results clearly showed an improved biocontrol effect of the genetically-modified strains against B. bruxellensis when compared with the modified Ethanol-Red strain with the empty plasmid (without the AMPs-codifying genes) and with the non-modified Ethanol-Red strain. In mixed-culture fermentation with the modified S. cerevisiae strain, B. bruxellensis culturability decreased from 5×104 CFU/mL on day-0 to less than 1 CFU/mL on day-10, while in single-culture B. bruxellensis increased its culturability from 6×104 to 1×106 CFU/mL in the first 6 days and kept this value until day-10. Besides, the modified Ethanol-Red strain exhibited an enhanced antagonistic effect against B. bruxellensis when compared with that induced by the non-modified Ethanol-Red strain. Indeed, culturability loss of B. bruxellensis after 10 days of fermentation with the modified Ethanol-Red strain was 98.7 and 100% higher than that occurred in fermentations performed with the non-modified Ethanol-Red and the empty-plasmid modified strain, respectively. Therefore, one can conclude that the S. cerevisiae genetically modified strain obtained in the present work may be a valuable solution for the mitigation of microbial contamination in fuel-ethanol fermentations, representing a much safer and environmentally friendly preservation strategy than the antimicrobial treatments (acid washing and antibiotics) currently applied in fuel-ethanol industry.Keywords: antimicrobial peptides, fuel-ethanol microbial contaminations, fuel-ethanol fermentation, biocontrol agents, genetically-modified yeasts
Procedia PDF Downloads 991531 Ruminal VFA of Beef Fed Different Protein
Authors: P. Paengkoum, S. C. Chen, S. Paengkoum
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Six male growing Thai-indigenous beef cattle with body weight (BW) of 154±13.2 kg were randomly assigned in replicated 3×3 Latin square design, and fed with different levels of crude protein (CP) in total mixed ration (TMR) diets. CP levels in diets were 4.3%, 7.3% and 10.3% base on dry matter (DM). Ruminal ammonia nitrogen (NH3-N) and blood urea nitrogen (BUN) concentrations increased (P<0.01) with increasing CP levels. Moreover, there is a positive relationship between BUN and ruminal NH3-N. Rumen pH, total volatile fatty acid (VFA), molar proportions of acetate, propionate and butyrate were not affected by CP levels (P>0.05).Keywords: Thai-indigenous beef cattle, crude protein, volatile fatty acid (VFA), total mixed ration (TMR) diets
Procedia PDF Downloads 2811530 Genome Sequencing of the Yeast Saccharomyces cerevisiae Strain 202-3
Authors: Yina A. Cifuentes Triana, Andrés M. Pinzón Velásco, Marío E. Velásquez Lozano
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In this work the sequencing and genome characterization of a natural isolate of Saccharomyces cerevisiae yeast (strain 202-3), identified with potential for the production of second generation ethanol from sugarcane bagasse hydrolysates is presented. This strain was selected because its capability to consume xylose during the fermentation of sugarcane bagasse hydrolysates, taking into account that many strains of S. cerevisiae are incapable of processing this sugar. This advantage and other prominent positive aspects during fermentation profiles evaluated in bagasse hydrolysates made the strain 202-3 a candidate strain to improve the production of second-generation ethanol, which was proposed as a first step to study the strain at the genomic level. The molecular characterization was carried out by genome sequencing with the Illumina HiSeq 2000 platform paired end; the assembly was performed with different programs, finally choosing the assembler ABYSS with kmer 89. Gene prediction was developed with the approach of hidden Markov models with Augustus. The genes identified were scored based on similarity with public databases of nucleotide and protein. Records were organized from ontological functions at different hierarchical levels, which identified central metabolic functions and roles of the S. cerevisiae strain 202-3, highlighting the presence of four possible new proteins, two of them probably associated with the positive consumption of xylose.Keywords: cellulosic ethanol, Saccharomyces cerevisiae, genome sequencing, xylose consumption
Procedia PDF Downloads 3201529 Screening of High-Alcohol Producing Yeasts for Manufacturing Process of Whisky
Authors: Byeong-Uk Lim, Young-Ran Song, Sang-Ho Baik
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This study aimed to develop yeast starters for scientific alcohol production and systematic quality control of whisky. A total of 389 yeast strains were isolated from traditional Korean fermentation starter (nuruk) and rice wine (makgeolli), and ten strains were finally selected for their high alcohol productivities, in which their alcohol productions were above 17.3% (v/v) during 10 days under two steps of glucose feeding condition (30% and then 15%, w/v). By 18s rDNA sequence analysis, all strains were identified as Saccharomyces cerevisiae (SC), and they can grow well under 50% (w/v) glucose and 10% (v/v) ethanol conditions. Furthermore, the capability of ten different SC strains to ferment rice wine for whisky was studied. Rice wine was fermented with only steamed rice, water, and two types of enzymes (glucoamylase and α-amylase) during 14 days at 25 °C, and then their oenological properties have been determined. As the results, the fermented rice wines indicated the final pH range of 4.24-4.38 and acidity range of 0.12-0.18. The highest ethanol production of 20.2% (v/v) was found in the fermentation with a SC-156 strain, whereas SC-92 (16.8%) and SC-119 (16.4%) showed significantly lowest ethanol productions. In addition, the residual sugar contents showed negative correlation with alcohol contents. Moreover, this study focused on nucleotide polymorphisms in the MSN2 and MSN4 genes to investigate the cause of the defective stress responses in yeast. Consequently, it was also confirmed that the deletion of the N termini of Msn4p from identified point mutations in SC-63, SC-95, SC-156, SC-158, and SC-160 strains.Keywords: yeast, high-alcohol, whisky, rice wine
Procedia PDF Downloads 3231528 Antidiabetic and Admet Pharmacokinetic Properties of Grewia Lasiocarpa E. Mey. Ex Harv. Stem Bark Extracts: An in Vitro and in Silico Study
Authors: Akwu N. A., Naidoo Y., Salau V. F., Olofinsan K. A.
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Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is a Southern African medicinal plant indigenously used with other plants for birthing problems. The anti-diabetic properties of the hexane, chloroform, and methanol extracts of Grewia lasiocarpa stem bark were assessed using in vitro α-glucosidase enzyme inhibition assay. The predictive in silico drug-likeness and toxicity properties of the phytocompounds were conducted using the pKCSM, ADMElab, and SwissADME computer-aided online tools. The highest α-glucosidase percentage inhibition was observed in the hexane extract (86.76%, IC50= 0.24 mg/mL), followed by chloroform (63.08%, IC50= 4.87 mg/mL) and methanol (53.22%, IC50= 9.41 mg/mL); while acarbose, the standard anti-diabetic drug was (84.54%, IC50= 1.96 mg/mL). The α-glucosidase assay revealed that the hexane extract exhibited the strongest carbohydrate inhibiting capacity and is a better inhibitor than the standard reference drug-acarbose. The computational studies also affirm the results observed in the in vitroα-glucosidaseassay. Thus, the extracts of G. lasiocarpa may be considered a potential plant-sourced compound for treating type 2 diabetes mellitus. This is the first study on the anti-diabetic properties of Grewia lasiocarpa hexane, chloroform, and methanol extracts using in vitro and in silico models.Keywords: grewia lasiocarpa, α-glucosidase inhibition, anti-diabetes, ADMET
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