Abstracts | Biotechnology and Bioengineering
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1021

World Academy of Science, Engineering and Technology

[Biotechnology and Bioengineering]

Online ISSN : 1307-6892

301 Developing a Process and Cost Model for Xanthan Biosynthesis from Bioethanol Production Waste Effluents

Authors: Bojana Ž. Bajić, Damjan G. Vučurović, Siniša N. Dodić, Jovana A. Grahovac, Jelena M. Dodić

Abstract:

Biosynthesis of xanthan, a microbial polysaccharide produced by Xanthomonas campestris, is characterized by the possibility of using non-specific carbohydrate substrates, which means different waste effluents can be used as a basis for the production media. Potential raw material sources for xanthan production come from industries with large amounts of waste effluents that are rich in compounds necessary for microorganism growth and multiplication. Taking into account the amount of waste effluents generated by the bioethanol industry and the fact that it contains a high inorganic and organic load it is clear that they represent a potential environmental pollutants if not properly treated. For this reason, it is necessary to develop new technologies which use wastes and wastewaters of one industry as raw materials for another industry. The result is not only a new product, but also reduction of pollution and environmental protection. Biotechnological production of xanthan, which consists of using biocatalysts to convert the bioethanol waste effluents into a high-value product, presents a possibility for sustainable development. This research uses scientific software developed for the modeling of biotechnological processes in order to design a xanthan production plant from bioethanol production waste effluents as raw material. The model was developed using SuperPro Designer® by using input data such as the composition of raw materials and products, defining unit operations, utility consumptions, etc., while obtaining capital and operating costs and the revenues from products to create a baseline production plant model. Results from this baseline model can help in the development of novel biopolymer production technologies. Additionally, a detailed economic analysis showed that this process for converting waste effluents into a high value product is economically viable. Therefore, the proposed model represents a useful tool for scaling up the process from the laboratory or pilot plant to a working industrial scale plant.

Keywords: biotechnology, process model, xanthan, waste effluents

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300 Synthesis of Double Dye-Doped Silica Nanoparticles and Its Application in Paper-Based Chromatography

Authors: Ka Ho Yau, Jan Frederick Engels, Kwok Kei Lai, Reinhard Renneberg

Abstract:

Lateral flow test is a prevalent technology in various sectors such as food, pharmacology and biomedical sciences. Colloidal gold (CG) is widely used as the signalling molecule because of the ease of synthesis, bimolecular conjugation and its red colour due to intrinsic SPRE. However, the production of colloidal gold is costly and requires vigorous conditions. The stability of colloidal gold are easily affected by environmental factors such as pH, high salt content etc. Silica nanoparticles are well known for its ease of production and stability over a wide range of solvents. Using reverse micro-emulsion (w/o), silica nanoparticles with different sizes can be produced precisely by controlling the amount of water. By incorporating different water-soluble dyes, a rainbow colour of the silica nanoparticles could be produced. Conjugation with biomolecules such as antibodies can be achieved after surface modification of the silica nanoparticles with organosilane. The optimum amount of the antibodies to be labelled was determined by Bradford Assay. In this work, we have demonstrated the ability of the dye-doped silica nanoparticles as a signalling molecule in lateral flow test, which showed a semi-quantitative measurement of the analyte. The image was further analysed for the LOD=10 ng of the analyte. The working range and the linear range of the test were from 0 to 2.15μg/mL and from 0 to 1.07 μg/mL (R2=0.988) respectively. The performance of the tests was comparable to those using colloidal gold with the advantages of lower cost, enhanced stability and having a wide spectrum of colours. The positives lines can be imaged by naked eye or by using a mobile phone camera for a better quantification. Further research has been carried out in multicolour detection of different biomarkers simultaneously. The preliminary results were promising as there was little cross-reactivity being observed for an optimized system. This approach provides a platform for multicolour detection for a set of biomarkers that enhances the accuracy of diseases diagnostics.

Keywords: colorimetric detection, immunosensor, paper-based biosensor, silica

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299 Characterization of Fatty Acid Glucose Esters as Os9BGlu31 Transglucosidase Substrates in Rice

Authors: Juthamath Komvongsa, Bancha Mahong, Kannika Phasai, Sukanya Luang, Jong-Seong Jeon, James Ketudat-Cairns

Abstract:

Os9BGlu31 is a rice transglucosidase that transfers glucosyl moieties to various acceptors such as carboxylic acids and alcohols, including phenolic acids and flavonoids, in vitro. The role of Os9BGlu31 transglucosidase in rice plant metabolism has not been reported to date. Methanolic extracts of rice bran and flag leaves were found to contain substrates to which Os9BGlu31 could transfer glucose from 4-nitrophenyl β -D-glucopyranoside donor. The semi-purified substrate from rice bran was found to contain oleic acid and linoleic acid and the pure fatty acids were found to act as acceptor substrates for Os9BGlu31 transglucosidase to form 1-O-acyl glucose esters. Os9BGlu31 showed higher activity with oleic acid (18:1) and linoleic acid (18:2) than stearic acid (18:0), and had both higher kcat and higher Km for linoleic than oleic acid in the presence of 8 mM 4NPGlc donor. This transglucosidase reaction is reversible, Os9bglu31 knockout rice lines of flag leaves were found to have higher amounts of fatty acid glucose esters than wild type control lines, these data conclude that fatty acid glucose esters act as glucosyl donor substrates for Os9BGlu31 transglucosidase in rice.

Keywords: fatty acid, fatty acid glucose ester, transglucosidase, rice flag leaf, homologous knockout lines, tandam mass spectrometry

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298 Genetic Change in Escherichia coli KJ122 That Improved Succinate Production from an Equal Mixture of Xylose and Glucose

Authors: Apichai Sawisit, Sirima Suvarnakuta Jantama, Sunthorn Kanchanatawee, Lonnie O. Ingram, Kaemwich Jantama

Abstract:

Escherichia coli KJ122 was engineered to produce succinate from glucose using the wild type GalP for glucose uptake instead of the native phosphotransferase system (ptsI mutation). This strain ferments 10% (w/v) xylose poorly. Mutants were selected by serial transfers in AM1 mineral salts medium with 10% (w/v) xylose. Evolved mutants exhibited a similar improvement, co-fermentation of an equal mixture of xylose and glucose. One of these, AS1600a, produced 84.26±1.37 g/L succinate, equivalent to that produced by the parent (KJ122) strain from 10% glucose (85.46±1.78 g/L). AS1600a was sequenced and found to contain a mutation in galactose permease (GalP, G236D). Expressing the galP* mutation gene in KJ122ΔgalP resembled the xylose utilization phenotype of the mutant AS1600a. The strain AS1600a and KJ122ΔgalP (pLOI5746; galP*) also co-fermented a mixture of glucose, xylose, arabinose, and galactose in sugarcane bagasse hydrolysate for succinate production.

Keywords: xylose, furfural, succinate, sugarcane bagasse, E. coli

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297 Fat-Tail Test of Regulatory DNA Sequences

Authors: Jian-Jun Shu

Abstract:

The statistical properties of CRMs are explored by estimating similar-word set occurrence distribution. It is observed that CRMs tend to have a fat-tail distribution for similar-word set occurrence. Thus, the fat-tail test with two fatness coefficients is proposed to distinguish CRMs from non-CRMs, especially from exons. For the first fatness coefficient, the separation accuracy between CRMs and exons is increased as compared with the existing content-based CRM prediction method – fluffy-tail test. For the second fatness coefficient, the computing time is reduced as compared with fluffy-tail test, making it very suitable for long sequences and large data-base analysis in the post-genome time. Moreover, these indexes may be used to predict the CRMs which have not yet been observed experimentally. This can serve as a valuable filtering process for experiment.

Keywords: statistical approach, transcription factor binding sites, cis-regulatory modules, DNA sequences

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296 Isolation and Screening of Laccase Producing Basidiomycetes via Submerged Fermentations

Authors: Mun Yee Chan, Sin Ming Goh, Lisa Gaik Ai Ong

Abstract:

Approximately 10,000 different types of dyes and pigments are being used in various industrial applications yearly, which include the textile and printing industries. However, these dyes are difficult to degrade naturally once they enter the aquatic system. Their high persistency in natural environment poses a potential health hazard to all form of life. Hence, there is a need for alternative dye removal strategy in the environment via bioremediation. In this study, fungi laccase is investigated via commercial agar dyes plates and submerged fermentation to explore the application of fungi laccase in textile dye wastewater treatment. Two locally isolated basidiomycetes were screened for laccase activity using media added with commercial dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3 (1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS plates with the appearance of greenish halo on around the isolates. Submerged fermentation performed on Isolate TBB3 with the productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed higher laccase production, it was subjected to molecular characterization by DNA isolation, PCR amplification and sequencing of ITS region of nuclear ribosomal DNA. After being compared with other sequences in National Center for Biotechnology Information (NCBI database), isolate TBB3 is probably from species Trametes hirsutei. Further research work can be performed on this isolate by upscale the production of laccase in order to meet the demands of the requirement for higher enzyme titer for the bioremediation of textile dyes.

Keywords: bioremediation, dyes, fermentation, laccase

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295 Prediction and Identification of a Permissive Epitope Insertion Site for St Toxoid in cfaB from Enterotoxigenic Escherichia coli

Authors: N. Zeinalzadeh, Mahdi Sadeghi

Abstract:

Enterotoxigenic Escherichia coli (ETEC) is the most common cause of non-inflammatory diarrhea in the developing countries, resulting in approximately 20% of all diarrheal episodes in children in these areas. ST is one of the most important virulence factors and CFA/I is one of the frequent colonization factors that help to process of ETEC infection. ST and CfaB (CFA/I subunit) are among vaccine candidates against ETEC. So, ST because of its small size is not a good immunogenic in the natural form. However to increase its immunogenic potential, here we explored candidate positions for ST insertion in CfaB sequence. After bioinformatics analysis, one of the candidate positions was selected and the chimeric gene (cfaB*st) sequence was synthesized and expressed in E. coli BL21 (DE3). The chimeric recombinant protein was purified with Ni-NTA columns and characterized with western blot analysis. The residue 74-75 of CfaB sequence could be a good candidate position for ST and other epitopes insertion.

Keywords: bioinformatics, CFA/I, enterotoxigenic E. coli, ST toxoid

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294 Detection of Nanotoxic Material Using DNA Based QCM

Authors: Juneseok You, Chanho Park, Kuehwan Jang, Sungsoo Na

Abstract:

Sensing of nanotoxic materials is strongly important, as their engineering applications are growing recently and results in that nanotoxic material can harmfully influence human health and environment. In current study we report the quartz crystal microbalance (QCM)-based, in situ and real-time sensing of nanotoxic-material by frequency shift. We propose the in situ detection of nanotoxic material of zinc oxice by using QCM functionalized with a taget-specific DNA. Since the mass of a target material is comparable to that of an atom, the mass change caused by target binding to DNA on the quartz electrode is so small that it is practically difficult to detect the ions at low concentrations. In our study, we have demonstrated the in-situ and fast detection of zinc oxide using the quartz crystal microbalance (QCM). The detection was derived from the DNA hybridization between the DNA on the quartz electrode. The results suggest that QCM-based detection opens a new avenue for the development of a practical water-testing sensor.

Keywords: nanotoxic material, qcm, frequency, in situ sensing

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293 Isolation and Characterization of an Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation

Authors: Rukshika S Hewawasam, Sisira K. Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran

Abstract:

Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. So inventions in microbes, substrates, technologies in fermentation cause new modifications in fermentation. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as pH, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance, growth curve assay were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 40% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test ,Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test. Positive results for Oxidase test was shown by isolated bacterium. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; pH 4, temperature 30˚C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.0±0.6% (V/V). Methanol was not detected in the final product of the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium

Keywords: bacterium, bio-fuel, ethanol tolerance, fermentation

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292 Adsorption of Heavy Metals Using Chemically-Modified Tea Leaves

Authors: Phillip Ahn, Bryan Kim

Abstract:

Copper is perhaps the most prevalent heavy metal used in the manufacturing industries, from food additives to metal-mechanic factories. Common methodologies to remove copper are expensive and produce undesired by-products. A good decontaminating candidate should be environment-friendly, inexpensive, and capable of eliminating low concentrations of the metal. This work suggests chemically modified spent tea leaves of chamomile, peppermint and green tea in their thiolated, sulfonated and carboxylated forms as candidates for the removal of copper from solutions. Batch experiments were conducted to maximize the adsorption of copper (II) ions. Effects such as acidity, salinity, adsorbent dose, metal concentration, and presence of surfactant were explored. Experimental data show that maximum adsorption is reached at neutral pH. The results indicate that Cu(II) can be removed up to 53%, 22% and 19% with the thiolated, carboxylated and sulfonated adsorbents, respectively. Maximum adsorption of copper on TPM (53%) is achieved with 150 mg and decreases with the presence of salts and surfactants. Conversely, sulfonated and carboxylated adsorbents show better adsorption in the presence of surfactants. Time-dependent experiments show that adsorption is reached in less than 25 min for TCM and 5 min for SCM. Instrumental analyses determined the presence of active functional groups, thermal resistance, and scanning electron microscopy, indicating that both adsorbents are promising materials for the selective recovery and treatment of metal ions from wastewaters. Finally, columns were prepared with these adsorbents to explore their application in scaled-up processes, with very positive results. A long-term goal involves the recycling of the exhausted adsorbent and/or their use in the preparation of biofuels due to changes in materials’ structures.

Keywords: heavy metal removal, adsorption, wastewaters, water remediation

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291 Bacterial Cellulose: A New Generation Antimicrobial Wound Dressing Biomaterial

Authors: Bhavana V. Mohite, Satish V. Patil

Abstract:

Bacterial cellulose (BC) is an alternative for plant cellulose (PC) that prevents global warming leads to preservation of nature. Although PC and BC have the same chemical structure, BC is superior with its properties like its size, purity, porosity, degree of polymerization, crystallinity and water holding capacity, thermal stability etc. On this background the present study focus production and applications of BC as antimicrobial wound dressing material. BC was produced by Gluconoacetobacter hansenii (strain NCIM 2529) under shaking condition and statistically enhanced upto 7.2 g/l from 3.0 g/l. BC was analyzed for its physico mechanical, structural and thermal characteristics. BC produced at shaking condition exhibits more suitable properties in support to its high performance applications. The potential of nano silver impregnated BC was determined for sustained release modern antimicrobial wound dressing material by swelling ratio, mechanical properties and antimicrobial activity against Staphylococcus aureus. BC in nanocomposite form with other synthetic polymer like PVA shows improvement in its properties such as swelling ratio (757% to 979%) and sustainable release of antibacterial agent. The high drug loading and release potential of BC was evidenced in support to its nature as antimicrobial wound dressing material. The nontoxic biocompatible nature of BC was confirmed by MTT assay on human epidermal cells with 90% cell viability that allows its application as a regenerative biomaterial. Thus, BC as a promising new generation antimicrobial wound dressing material was projected.

Keywords: agitated culture, biopolymer, gluconoacetobacter hansenii, nanocomposite

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290 Fluoride-Induced Stress and Its Association with Bone Developmental Pathway in Osteosarcoma Cells

Authors: Deepa Gandhi, Pravin K. Naoghare, Amit Bafana, Krishnamurthi Kannan, Saravanadevi Sivanesana

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Oxidative stress is known to depreciate normal functioning of osteoblast cells. Present study reports oxidative/inflammatory signatures in fluoride exposed human osteosarcoma (HOS) cells and its possible association with the genes involved in bone developmental pathway. Microarray analysis was performed to understand the possible molecular mechanisms of stress-mediated bone lose in HOS cells. Cells were chronically exposed with sub-lethal concentration of fluoride. Global gene expression is profiling revealed 34 up regulated and 2598 down-regulated genes, which were associated with several biological processes including bone development, osteoblast differentiation, stress response, inflammatory response, apoptosis, regulation of cell proliferation. Microarray data were further validated through qRT-PCR and western blot analyses using key representative genes. Based on these findings, it can be proposed that chronic exposure of fluoride may impair bone development via oxidative and inflammatory stress. The present finding also provides important biological clues, which will be helpful for the development of therapeutic targets against diseases related bone.

Keywords: bone, HOS cells, microarray, stress

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289 Automated Human Balance Assessment Using Contactless Sensors

Authors: Justin Tang

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Balance tests are frequently used to diagnose concussions on the sidelines of sporting events. Manual scoring, however, is labor intensive and subjective, and many concussions go undetected. This study institutes a novel approach to conducting the Balance Error Scoring System (BESS) more quantitatively using Microsoft’s gaming system Kinect, which uses a contactless sensor and several cameras to receive data and estimate body limb positions. Using a machine learning approach, Visual Gesture Builder, and a deterministic approach, MATLAB, we tested whether the Kinect can differentiate between “correct” and erroneous stances of the BESS. We created the two separate solutions by recording test videos to teach the Kinect correct stances and by developing a code using Java. Twenty-two subjects were asked to perform a series of BESS tests while the Kinect was collecting data. The Kinect recorded the subjects and mapped key joints onto their bodies to obtain angles and measurements that are interpreted by the software. Through VGB and MATLAB, the videos are analyzed to enumerate the number of errors committed during testing. The resulting statistics demonstrate a high correlation between manual scoring and the Kinect approaches, indicating the viability of the use of remote tracking devices in conducting concussion tests.

Keywords: automated, concussion detection, contactless sensors, microsoft kinect

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288 Cell Elevator: A Novel Technique for Cell Sorting and Circulating Tumor Cell Detection and Discrimination

Authors: Kevin Zhao, Norman J. Horing

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A methodology for cells sorting and circulating tumor cell detection and discrimination is presented in this paper. The technique is based on Dielectrophoresis and microfluidic device theory. Specifically, the sorting of the cells is realized by adjusting the relation among the sedimentation forces, the drag force provided by the fluid, and the Dielectrophortic force that is relevant to the bias voltage applied on the device. The relation leads to manipulation of the elevation of the cells of the same kind to a height by controlling the bias voltage. Once the cells have been lifted to a position next to the bottom of the cell collection channel, the buffer fluid flashes them into the cell collection channel. Repeated elevation of the cells leads to a complete sorting of the cells in the sample chamber. A proof-of-principle example is presented which verifies the feasibility of the methodology.

Keywords: cell sorter, CTC cell, detection and discrimination, dielectrophoresisords, simulation

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287 Process Development for the Conversion of Organic Waste into Valuable Products

Authors: Ife O. Bolaji

Abstract:

Environmental concerns arising from the use of fossil fuels has increased the interest in the development of renewable and sustainable sources of energy. This would minimize the dependence on fossil fuels and serve as future alternatives. Organic wastes contain carbohydrates, proteins and lipids, which can be utilised as carbon sources for the production of bio-based products. Cellulose is the most abundant natural biopolymer, being the main structural component of lignocellulosic materials. The aim of this project is to develop a biological process for the hydrolysis and fermentation of organic wastes into ethanol and organic acids. The hydrolysis and fermentation processes are integrated in a single vessel using undefined mixed culture microorganisms. The anaerobic fermentation of microcrystalline cellulose was investigated in continuous and batch reactors at 25°C with an appropriate growth medium for cellulase formation, hydrolysis, and fermentation. The reactors were inoculated with soil (B1, C1, C3) or sludge from an anaerobic digester (B2, C2) and the breakdown of cellulose was monitored by measuring the production of ethanol, organic acids and the residual cellulose. The batch reactors B1 and B2 showed negligible microbial activity due to inhibition while the continuous reactors, C1, C2 and C3, exhibited little cellulose hydrolysis which was concealed by the cellulose accumulation in the reactor. At the end of the continuous operation, the reactors C1, C2 and C3 were operated under batch conditions. 48%, 34% and 42% cellulose had been fermented by day 88, 55 and 55 respectively of the batch fermentation. Acetic acid, ethanol, propionic acid and butyric acids were the main fermentation products in the reactors. A stable concentration of 0.6 g/l ethanol and 5 g/L acetic acid was maintained in C3 for several weeks due to reduced activity of methanogens caused by the decrease in pH. Thus far, the results have demonstrated that mixed microbial culture is capable of hydrolysing and fermenting cellulose under lenient conditions. The fermentation of cellulose has been found effective in a combination of continuous and batch processes.

Keywords: cellulose, hydrolysis, mixed culture, organic waste

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286 Production and Characterisation of Lipase from a Novel Streptomyces.sp - Its Molecular Identification

Authors: C. Asha Poorna, N. S. Pradeep

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The biological function of lipase is to catalyze the hydrolysis of triacylglycerols to give free fatty acid, diacylglycerols, mono-acylglycerols and glycerol. They constitute the most important group of biocatalysts for biotechnological applications. The aim of the present study was to identify the lipolytic activity of Streptomyces sp. From soil sample collected from the sacred groves of southern Kerala. The culture conditions of the isolate were optimised and the enzyme was purified and characterised. The purification was attempted with acetone precipitation. The isolate observed to have high lipolytic activity and identified to be of Streptomyces strain. The purification was attempted with acetone precipitation. The purified enzyme observed to have an apparent molecular mass of ~60kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme showed maximum activity at 60oC and pH-8. The lipase showed tolerance towards different organic solvents like ethanol and methanol that are commonly used in transesterification reactions to displace alcohol from triglycerides contained in renewable resources to yield fatty acid alkyl esters known as biodiesel.

Keywords: lipase, Streptomyces, biodiesel, fatty acid, transesterification

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285 Review of Hydrologic Applications of Conceptual Models for Precipitation-Runoff Process

Authors: Oluwatosin Olofintoye, Josiah Adeyemo, Gbemileke Shomade

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The relationship between rainfall and runoff is an important issue in surface water hydrology therefore the understanding and development of accurate rainfall-runoff models and their applications in water resources planning, management and operation are of paramount importance in hydrological studies. This paper reviews some of the previous works on the rainfall-runoff process modeling. The hydrologic applications of conceptual models and artificial neural networks (ANNs) for the precipitation-runoff process modeling were studied. Gradient training methods such as error back-propagation (BP) and evolutionary algorithms (EAs) are discussed in relation to the training of artificial neural networks and it is shown that application of EAs to artificial neural networks training could be an alternative to other training methods. Therefore, further research interest to exploit the abundant expert knowledge in the area of artificial intelligence for the solution of hydrologic and water resources planning and management problems is needed.

Keywords: artificial intelligence, artificial neural networks, evolutionary algorithms, gradient training method, rainfall-runoff model

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284 Isolate-Specific Variations among Clinical Isolates of Brucella Identified by Whole-Genome Sequencing, Bioinformatics and Comparative Genomics

Authors: Abu S. Mustafa, Mohammad W. Khan, Faraz Shaheed Khan, Nazima Habibi

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Brucellosis is a zoonotic disease of worldwide prevalence. There are at least four species and several strains of Brucella that cause human disease. Brucella genomes have very limited variation across strains, which hinder strain identification using classical molecular techniques, including PCR and 16 S rDNA sequencing. The aim of this study was to perform whole genome sequencing of clinical isolates of Brucella and perform bioinformatics and comparative genomics analyses to determine the existence of genetic differences across the isolates of a single Brucella species and strain. The draft sequence data were generated from 15 clinical isolates of Brucella melitensis (biovar 2 strain 63/9) using MiSeq next generation sequencing platform. The generated reads were used for further assembly and analysis. All the analysis was performed using Bioinformatics work station (8 core i7 processor, 8GB RAM with Bio-Linux operating system). FastQC was used to determine the quality of reads and low quality reads were trimmed or eliminated using Fastx_trimmer. Assembly was done by using Velvet and ABySS softwares. The ordering of assembled contigs was performed by Mauve. An online server RAST was employed to annotate the contigs assembly. Annotated genomes were compared using Mauve and ACT tools. The QC score for DNA sequence data, generated by MiSeq, was higher than 30 for 80% of reads with more than 100x coverage, which suggested that data could be utilized for further analysis. However when analyzed by FastQC, quality of four reads was not good enough for creating a complete genome draft so remaining 11 samples were used for further analysis. The comparative genome analyses showed that despite sharing same gene sets, single nucleotide polymorphisms and insertions/deletions existed across different genomes, which provided a variable extent of diversity to these bacteria. In conclusion, the next generation sequencing, bioinformatics, and comparative genome analysis can be utilized to find variations (point mutations, insertions and deletions) across different genomes of Brucella within a single strain. This information could be useful in surveillance and epidemiological studies supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: brucella, bioinformatics, comparative genomics, whole genome sequencing

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283 From Orthodox to Haploid Mitochondrial DNA Markers: Exploring the Datum Folder of population of Sindh in Pakistan

Authors: Shahzad Bhattiab, M. Aslamkhana, Sana Abbasbc, Marcella Attimonellid, Kumarasamy Thangaraje, Erica Martinha Silva de Souzaf, Uzay U. Sezen

Abstract:

The present study was designed to investigate three regions of mitochondrial DNA, HVI, HVII and HVIII, to hold a powwow genetic diversity and affiliations in 115 probands of 6 major ethnic groups, viz., Bijarani, Chandio, Ghallu, Khoso, Nasrani and Solangi, in the province of Sindh of Pakistan. For this purpose 88 haplotypes were scrutinized, defined by particular set of nucleotides (ignoring the C insertions around position 309 and 315). In spite of that 82% sequences were observed once, 12 % twice and 5.2 % thrice. The most common South Asian haplotypes were observed M (42%), N (6.9%) and R (6.9%) whereas west Eurasian haplotypes were J (1.7%), U (23.4%), H (9.5%), W (6.9%) and T (0.86%), in six ethnic groups. A random match probability between two unrelated individuals was found 0.06 %, while genetic diversity was ranged to be 0.991 to 0.999, and nucleotide diversity ranged from 0.0089 to 0.0142 for the whole control region of the population studied.

Keywords: mtDNA haplogroups, control region, Pakistan, Sindh, ethnicity

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282 Biogas Production from University Canteen Waste: Effect of Organic Loading Rate and Retention Time

Authors: Khamdan Cahyari, Gumbolo Hadi Susanto, Pratikno Hidayat, Sukirman

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University canteen waste was used as raw material to produce biogas in Faculty of Industrial Technology, Islamic University of Indonesia. This faculty was home to more than 3000 students and lecturers who work and study for 5 days/week (8 hours/day). It produced approximately 85 ton/year organic fraction of canteen waste. Yet, this waste had been dumped for years in landfill area which cause severe environmental problems. It was proposed to utilize the waste as raw material for producing renewable energy source of biogas. This research activities was meant to investigate the effect of organic loading rate (OLR) and retention time (RT) of continuous anaerobic digestion process for 200 days. Organic loading rate was set at value 2, 3, 4 and 5 g VS/l/d whereas the retention time was adjusted at 30, 24, 18 and 14.4 days. Optimum condition was achieved at OLR 4 g VS/l/d and RT 24 days with biogas production rate between 0.75 to 1.25 liter/day (40-60% CH4). This indicated that the utilization of canteen waste to produce biogas was promising method to mitigate environmental problem of university canteen waste. Furthermore, biogas could be used as alternative energy source to supply energy demand at the university. This implementation is simultaneous solution for both waste and energy problems to achieve green campus.

Keywords: canteen waste, biogas, anaerobic digestion, university, green campus

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281 Effect of Interaction between Different Concentrations of Colchicine, Time Duration and Two Verities of Crepis capillaris on Chromosome Polyploidy in vitro Culture

Authors: Mosleh M. S. Duhoky, Payman A. A. Zibari

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These experiments were conducted at Tissue Culture Laboratory/ Faculty of Agriculture and Forestry/ University of Duhok during the period from January 2011 to May 2013. The objectives of this study were to study the effects of interaction between three different factors on percentage of polyploidy of Crepis capillaris by using Tissue culture technology. Concerning the data it is obvious that shaking of Crepis capillaris with 2B chromosome with 0.15 mM for ten days inscribed a high percentage of polyploidy within most fifteen passages.

Keywords: crepis capillaris, 2B chromosome, tissue culture, polyploidy

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280 Heritability and Diversity Analysis of Blast Resistant Upland Rice Genotypes Based on Quantitative Traits

Authors: Mst. Tuhina-Khatun, Mohamed Hanafi Musa, Mohd Rafii Yosup, Wong Mui Yun, Md. Aktar-Uz-Zaman, Mahbod Sahebi

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Rice is a staple crop of economic importance of most Asian people, and blast is the major constraints for its higher yield. Heritability of plants traits helps plant breeders to make an appropriate selection and to assess the magnitude of genetic improvement through hybridization. Diversity of crop plants is necessary to manage the continuing genetic erosion and address the issues of genetic conservation for successfully meet the future food requirements. Therefore, an experiment was conducted to estimate heritability and to determine the diversity of 27 blast resistant upland rice genotypes based on 18 quantitative traits using randomized complete block design. Heritability value was found to vary from 38 to 93%. The lowest heritability belonged to the character total number of tillers/plant (38%). In contrast, number of filled grains/panicle, and yield/plant (g) was recorded for their highest heritability value viz. 93 and 91% correspondingly. Cluster analysis based on 18 traits grouped 27 rice genotypes into six clusters. Cluster I was the biggest, which comprised 17 genotypes, accounted for about 62.96% of total population. The multivariate analysis suggested that the genotype ‘Chokoto 14’ could be hybridized with ‘IR 5533-55-1-11’ and ‘IR 5533-PP 854-1’ for broadening the gene pool of blast resistant upland rice germplasms for yield and other favorable characters.

Keywords: blast resistant, diversity analysis, heritability, upland rice

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279 Batch Adsorption Studies for the Removal of Textile Dyes from Aqueous Solution on Three Different Pine Bark

Authors: B. Cheknane, F. Zermane

Abstract:

The main objective of the present study is the valorization of natural raw materials of plant origin for the treatment of textile industry wastewater. Selected bark was: maritime (MP), pinyon (PP) and Aleppo pine (AP) bark. The efficiency of these barks were tested for the removal of three dye; rhodamine B (RhB), Green Malachite (GM) and X Methyl Orange (MO). At the first time we focus to study the different parameters which can influence the adsorption processes such as: nature of the adsorbents, nature of the pollutants (dyes) and the effect of pH. Obtained results reveals that the speed adsorption is strongly influencing by the pH medium and the comparative study show that adsorption is favorable in the acidic medium with amount adsorbed of (Q=40mg/g) for rhodamine B and (Q=46mg/g) for orange methyl. Results of adsorption kinetics reveals that the molecules of GM are adsorbed better (Q=48mg/g) than the molecules of RhB (Q=46mg/g) and methyl orange (Q=18mg/g), with equilibrium time of 6 hours. The results of adsorption isotherms show clearly that the maritime pine bark is the most effective adsorbents with adsorbed amount of (QRhB=200mg/g) and (QMO=88mg/g) followed by pinyon pine (PP) with (QRhB=184mg/g) and (QMO=56mg/g) and finally Aleppo pine (AP) bark with (QRhB=131mg/g) and (QMO= 46mg/g). The different obtained isotherms were modeled using the Langmuir and Freundlich models and according to the adjustment coefficient values R2, the obtained isotherms are well represented by Freundlich model.

Keywords: maritime pine bark (MP), pinyon pine bark (PP), Aleppo pine (AP) bark, adsorption, dyes

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278 Biodegradation of Direct Red 23 by Bacterial Consortium Isolated from Dye Contaminated Soil Using Sequential Air-lift Bioreactor

Authors: Lata Kumari Dhanesh Tiwary, Pradeep Kumar Mishra

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The effluent coming from various industries such as textile, carpet, food, pharmaceutical and many other industries is big challenge due to its recalcitrant and xenobiotiocs in nature. Recently, biodegradation of dye wastewater through biological means was widely used due to eco-friendly and cost effective with the higher percentage of removal of dye from wastewater. The present study deals with the biodegradation and decolourization of Direct Red 23 dye using indigenously isolated bacterial consortium. The bacterial consortium was isolated from soil sample from dye contaminated site near a cluster of Carpet industries of Bhadohi, Uttar Pradesh, India. The bacterial strain formed consortia were identified and characterized by morphological, biochemical and 16S rRNA gene sequence analysis. The bacterial strain mainly Staphylococcus saprophyticus strain BHUSS X3 (KJ439576), Microbacterium sp. BHUMSp X4 (KJ740222) and Staphylococcus saprophyticus strain BHUSS X5 (KJ439576) were used as consortia for further studies of dye decolorization. Experimental investigations were made in a Sequencing Air- lift bioreactor using the synthetic solution of Direct Red 23 dye by optimizing various parameters for efficient degradation of dye. The effect of several operating parameters such as flow rate, pH, temperature, initial dye concentration and inoculums size on removal of dye was investigated. The efficiency of isolated bacterial consortia from dye contaminated area in Sequencing Air- lift Bioreactor with different concentration of dye between 100-1200 mg/l at different hydraulic rate (HRTs) 26h and 10h. The maximum percentage of dye decolourization 98% was achieved when operated at HRT of 26h. The percentage of decolourization of dye was confirmed by using UV-Vis spectrophotometer and HPLC.

Keywords: carpet industry, bacterial consortia, sequencing air-lift bioreactor

Procedia PDF Downloads 322
277 Effect of Aeration on Bacterial Cellulose (BC) Production by Gluconacetobacter xylinus DSM46604 in Batch Fermentation

Authors: Azila Adnan, Giridhar R. Nair, Mark C. Lay, Janis E. Swan

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The effect of aeration on bacterial cellulose (BC) production by Gluconacetobacter xylinus DSM46604 was studied in 5-L bioreactor. Four aeration rates were applied (0.3, 0.6, 1.0 and 1.5 vvm) in the fermentation media at constant agitation rate, 150 rpm. One vvm enhanced BC concentration (4.4 g/L) and productivity (0.44 g/L/day) while greater agitation rate (1.5 vvm) decreased BC concentration (4.0 g/L) and productivity (0.40 g/L/day).

Keywords: aeration, bacterial cellulose, fermentation, gluconacetobacter xylinus

Procedia PDF Downloads 411
276 Stability and Rheology of Sodium Diclofenac-Loaded and Unloaded Palm Kernel Oil Esters Nanoemulsion Systems

Authors: Malahat Rezaee, Mahiran Basri, Raja Noor Zaliha Raja Abdul Rahman, Abu Bakar Salleh

Abstract:

Sodium diclofenac is one of the most commonly used drugs of nonsteroidal anti-inflammatory drugs (NSAIDs). It is especially effective in the controlling the severe conditions of inflammation and pain, musculoskeletal disorders, arthritis, and dysmenorrhea. Formulation as nanoemulsions is one of the nanoscience approaches that have been progressively considered in pharmaceutical science for transdermal delivery of drug. Nanoemulsions are a type of emulsion with particle sizes ranging from 20 nm to 200 nm. An emulsion is formed by the dispersion of one liquid, usually the oil phase in another immiscible liquid, water phase that is stabilized using surfactant. Palm kernel oil esters (PKOEs), in comparison to other oils; contain higher amounts of shorter chain esters, which suitable to be applied in micro and nanoemulsion systems as a carrier for actives, with excellent wetting behavior without the oily feeling. This research was aimed to study the effect of O/S ratio on stability and rheological behavior of sodium diclofenac loaded and unloaded palm kernel oil esters nanoemulsion systems. The effect of different O/S ratio of 0.25, 0.50, 0.75, 1.00 and 1.25 on stability of the drug-loaded and unloaded nanoemulsion formulations was evaluated by centrifugation, freeze-thaw cycle and storage stability tests. Lecithin and cremophor EL were used as surfactant. The stability of the prepared nanoemulsion formulations was assessed based on the change in zeta potential and droplet size as a function of time. Instability mechanisms including coalescence and Ostwald ripening for the nanoemulsion system were discussed. In comparison between drug-loaded and unloaded nanoemulsion formulations, drug-loaded formulations represented smaller particle size and higher stability. In addition, the O/S ratio of 0.5 was found to be the best ratio of oil and surfactant for production of a nanoemulsion with the highest stability. The effect of O/S ratio on rheological properties of drug-loaded and unloaded nanoemulsion systems was studied by plotting the flow curves of shear stress (τ) and viscosity (η) as a function of shear rate (γ). The data were fitted to the Power Law model. The results showed that all nanoemulsion formulations exhibited non-Newtonian flow behaviour by displaying shear thinning behaviour. Viscosity and yield stress were also evaluated. The nanoemulsion formulation with the O/S ratio of 0.5 represented higher viscosity and K values. In addition, the sodium diclofenac loaded formulations had more viscosity and higher yield stress than drug-unloaded formulations.

Keywords: nanoemulsions, palm kernel oil esters, sodium diclofenac, rheoligy, stability

Procedia PDF Downloads 407
275 Induction of Apoptosis by Diosmin through Interleukins/STAT and Mitochondria Mediated Pathway in Hep-2 and KB Cells

Authors: M. Rajasekar, K. Suresh

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Diosmin is a flavonoid, most abundantly found in many citrus fruits. As a flavonoid, it possesses a multitude of biological activities including anti-hyperglycemic, anti-lipid peroxidative, anti-inflammatory, antioxidant, and anti-mutagenic properties. At this point, we established the anti-proliferative and apoptosis-inducing activities of diosmin in Hep-2 and KB cells. Diosmin has cytotoxic effects through inhibiting cellular proliferation of Hep-2 and KB cells, which leads to the induction of apoptosis, as apparent by an increase in the fraction of cells in the sub-G1phase of the cell cycle. Results exposed that inhibition of cell proliferation is associated with regulation of the Interleukins/STAT pathway. In addition, Diosmin treatment with Hep-2 and KB cells actively stimulated reactive oxygen species (ROS) and mitochondrial membrane depolarization. And also an imbalance in the Bax/Bcl-2 ratio triggered the caspase cascade and shifting the balance in favor of apoptosis. These observations conclude that Diosmin induce apoptosis via Interleukins /STAT-mediated pathway.

Keywords: diosmin, apoptosis, antioxidant, STAT pathway

Procedia PDF Downloads 310
274 Chromium Reduction Using Bacteria: Bioremediation Technologies

Authors: Baljeet Singh Saharan

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Bioremediation is the demand of the day. Tannery and textile effluents/waste waters have lots of pollution due to presence of hexavalent Chromium. Methodologies used in the present investigations include isolation, cultivation and purification of bacterial strain. Further characterization techniques and 16S rRNA sequencing were performed. Efficient bacterial strain capable of reducing hexavalent chromium was obtained. The strain can be used for bioremediation of industrial effluents containing hexavalent Cr. A gram negative, rod shaped and yellowish pigment producing bacterial strain from tannery effluent was isolated using nutrient agar. The 16S rRNA gene sequence similarity indicated that isolate SA13A is associated with genus Luteimonas (99%). This isolate has been found to reduce 100% of hexavalent chromium Cr (VI) (100 mg L-1) 100% in 16 h. Growth conditions were optimized for Cr (VI) reduction. Maximum reduction was observed at a temperature of 37 °C and pH 8.0. Additionally, Luteimonas aestuarii SA13A showed resistance against various heavy metals like Cr+6, Cr+3, Cu+2, Zn+2, Co+2, Ni+2 and Cd+2 . Hence, Luteimonas aestuarii SA13A could be used as potent Cr (VI) reducing strain as well as significant bioremediator in heavy metal contaminated sites.

Keywords: bioremediation, chromium, eco-friendly, heavy metals

Procedia PDF Downloads 450
273 Comparison with Mechanical Behaviors of Mastication in Teeth Movement Cases

Authors: Jae-Yong Park, Yeo-Kyeong Lee, Hee-Sun Kim

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Purpose: This study aims at investigating the mechanical behaviors of mastication, according to various teeth movement. There are three masticatory cases which are general case and 2 cases of teeth movement. General case includes the common arrange of all teeth and 2 cases of teeth movement are that one is the half movement location case of molar teeth in no. 14 tooth seat after extraction of no. 14 tooth and the other is no. 14 tooth seat location case of molar teeth after extraction in the same case before. Materials and Methods: In order to analyze these cases, 3 dimensional finite element (FE) model of the skull were generated based on computed tomography images, 964 dicom files of 38 year old male having normal occlusion status. An FE model in general occlusal case was used to develop CAE procedure. This procedure was applied to FE models in other occlusal cases. The displacement controls according to loading condition were applied effectively to simulate occlusal behaviors in all cases. From the FE analyses, von Mises stress distribution of skull and teeth was observed. The von Mises stress, effective stress, had been widely used to determine the absolute stress value, regardless of stress direction and yield characteristics of materials. Results: High stress was distributed over the periodontal area of mandible under molar teeth when the mandible was transmitted to the coronal-apical direction in the general occlusal case. According to the stress propagation from teeth to cranium, stress distribution decreased as the distribution propagated from molar teeth to infratemporal crest of the greater wing of the sphenoid bone and lateral pterygoid plate in general case. In 2 cases of teeth movement, there were observed that high stresses were distributed over the periodontal area of mandible under teeth where they are located under the moved molar teeth in cranium. Conclusion: The predictions of the mechanical behaviors of general case and 2 cases of teeth movement during the masticatory process were investigated including qualitative validation. The displacement controls as the loading condition were applied effectively to simulate occlusal behaviors in 2 cases of teeth movement of molar teeth.

Keywords: cranium, finite element analysis, mandible, masticatory action, occlusal force

Procedia PDF Downloads 377
272 Noninvasive Disease Diagnosis through Breath Analysis Using DNA-functionalized SWNT Sensor Array

Authors: W. J. Zhang, Y. Q. Du, M. L. Wang

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Noninvasive diagnostics of diseases via breath analysis has attracted considerable scientific and clinical interest for many years and become more and more promising with the rapid advancement in nanotechnology and biotechnology. The volatile organic compounds (VOCs) in exhaled breath, which are mainly blood borne, particularly provide highly valuable information about individuals’ physiological and pathophysiological conditions. Additionally, breath analysis is noninvasive, real-time, painless and agreeable to patients. We have developed a wireless sensor array based on single-stranded DNA (ssDNA)-decorated single-walled carbon nanotubes (SWNT) for the detection of a number of physiological indicators in breath. Eight DNA sequences were used to functionalize SWNT sensors to detect trace amount of methanol, benzene, dimethyl sulfide, hydrogen sulfide, acetone and ethanol, which are indicators of heavy smoking, excessive drinking, and diseases such as lung cancer, breast cancer, cirrhosis and diabetes. Our tests indicated that DNA functionalized SWNT sensors exhibit great selectivity, sensitivity, reproducibility, and repeatability. Furthermore, different molecules can be distinguished through pattern recognition enabled by this sensor array. Thus, the DNA-SWNT sensor array has great potential to be applied in chemical or bimolecular detection for the noninvasive diagnostics of diseases and health monitoring.

Keywords: breath analysis, diagnosis, DNA-SWNT sensor array, noninvasive

Procedia PDF Downloads 333