Search results for: bacteria isolation
896 Bioinspired Green Synthesis of Magnetite Nanoparticles Using Room-Temperature Co-Precipitation: A Study of the Effect of Amine Additives on Particle Morphology in Fluidic Systems
Authors: Laura Norfolk, Georgina Zimbitas, Jan Sefcik, Sarah Staniland
Abstract:
Magnetite nanoparticles (MNP) have been an area of increasing research interest due to their extensive applications in industry, such as in carbon capture, water purification, and crucially, the biomedical industry. The use of MNP in the biomedical industry is rising, with studies on their effect as Magnetic resonance imaging contrast agents, drug delivery systems, and as hyperthermic cancer treatments becoming prevalent in the nanomaterial research community. Particles used for biomedical purposes must meet stringent criteria; the particles must have consistent shape and size between particles. Variation between particle morphology can drastically alter the effective surface area of the material, making it difficult to correctly dose particles that are not homogeneous. Particles of defined shape such as octahedral and cubic have been shown to outperform irregular shaped particles in some applications, leading to the need to synthesize particles of defined shape. In nature, highly homogeneous MNP are found within magnetotactic bacteria, a unique bacteria capable of producing magnetite nanoparticles internally under ambient conditions. Biomineralisation proteins control the properties of the MNPs, enhancing their homogeneity. One of these proteins, Mms6, has been successfully isolated and used in vitro as an additive in room-temperature co-precipitation reactions (RTCP) to produce particles of defined mono-dispersed size & morphology. When considering future industrial scale-up it is crucial to consider the costs and feasibility of an additive, as an additive that is not readily available or easily synthesized at a competitive price will not be sustainable. As such, additives selected for this research are inspired by the functional groups of biomineralisation proteins, but cost-effective, environmentally friendly, and compatible with scale-up. Diethylenetriamine (DETA), triethylenetetramine (TETA), tetraethylenepentamine (TEPA), and pentaethylenehexamine (PEHA) have been successfully used in RTCP to modulate the properties of particles synthesized, leading to the formation of octahedral nanoparticles with no use of organic solvents, heating, or toxic precursors. By extending this principle to a fluidic system, ongoing research will reveal whether the amine additives can also exert morphological control in an environment which is suited toward higher particle yield. Two fluidic systems have been employed; a peristaltic turbulent flow mixing system suitable for the rapid production of MNP, and a macrofluidic system for the synthesis of tailored nanomaterials under a laminar flow regime. The presence of the amine additives in the turbulent flow system in initial results appears to offer similar morphological control as observed under RTCP conditions, with higher proportions of octahedral particles formed. This is a proof of concept which may pave the way to green synthesis of tailored MNP on an industrial scale. Mms6 and amine additives have been used in the macrofluidic system, with Mms6 allowing magnetite to be synthesized at unfavourable ferric ratios, but no longer influencing particle size. This suggests this synthetic technique while still benefiting from the addition of additives, may not allow additives to fully influence the particles formed due to the faster timescale of reaction. The amine additives have been tested at various concentrations, the results of which will be discussed in this paper.Keywords: bioinspired, green synthesis, fluidic, magnetite, morphological control, scale-up
Procedia PDF Downloads 113895 Identification of Cellulose-Hydrolytic Thermophiles Isolated from Sg. Klah Hot Spring Based on 16S rDNA Gene Sequence
Authors: M. J. Norashirene, Y. Zakiah, S. Nurdiana, I. Nur Hilwani, M. H. Siti Khairiyah, M. J. Muhamad Arif
Abstract:
In this study, six bacterial isolates of a slightly thermophilic organism from the Sg. Klah hot spring, Malaysia were successfully isolated and designated as M7T55D1, M7T55D2, M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The bacterial isolates were screened for their cellulose hydrolytic ability on Carboxymethlycellulose agar medium. The isolated bacterial strains were identified morphologically, biochemically and molecularly with the aid of 16S rDNA sequencing. All of the bacteria showed their optimum growth at a slightly alkaline pH of 7.5 with a temperature of 55°C. All strains were Gram-negative, non-spore forming type, strictly aerobic, catalase-positive and oxidase-positive with the ability to produce thermostable cellulase. Based on BLASTn results, bacterial isolates of M7T55D2 and M7T53D1 gave the highest homology (97%) with similarity to Tepidimonas ignava while isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed their closest homology (97%-98%) with Tepidimonas thermarum. These cellulolytic thermophiles might have a commercial potential to produce valuable thermostable cellulase.Keywords: cellulase, cellulolytic, thermophiles, 16S rDNA gene
Procedia PDF Downloads 345894 Nematicidal Activity of the Cell Extract from Penicillium Sp EU0013 and Its Metabolite Profile Using High Performance Liquid Chromatograpy
Authors: Zafar Iqbal, Sana Irshad Khan
Abstract:
Organic extract from newly isolated plant growth promoting fungus (PGPF) Penicillium sp EU0013 was subjected to bioassays including anti fungal (disc diffusion) cytotoxicity (brine shrimp lethality), herbicidal (Lemna minor) and nematicidal activities. Metabolite profile of the extract was also assessed using HPLC analysis with the aim to identify bioactive natural products in the extract as new drug candidate(s). The extract showed anti fungal potential against tested fungal pathogens. Growth of the Wilt pathogen Fusarium oxyosproum was inhibited up to 63% when compared to negative reference. Activity against brine shrimps was weak and mortality up to 10% was observed at concentration of 200 µg. mL-1. The extract exhibited no toxicity against Lemna minor frond at 200 µg. mL-1. Nematicidal activity was observed very potent against root knot nematode and LC50 value was calculated as 52.5 ug. mL-1 using probit analysis. Methodically assessment of metabolites profile by HPLC showed the presence of kojic acid (Rt 1.4 min) and aflatoxin B1 (Rt 5.9 min) in the mycellial extract as compared with standards. The major unidentified metabolite was eluted at Rt 8.6 along with other minor peaks. The observed high toxicity against root knot nematode was attributed to the unidentified compounds that make fungal extract worthy of further exploration for isolation and structural characterization studies for development of future commercial nematicidal compound(s).Keywords: penicillium, nematicidal activity, metabolites, HPLC
Procedia PDF Downloads 446893 Aircraft Components, Manufacturing and Design: Opportunities, Bottlenecks, and Challenges
Authors: Ionel Botef
Abstract:
Aerospace products operate in very aggressive environments characterized by high temperature, high pressure, large stresses on individual components, the presence of oxidizing and corroding atmosphere, as well as internally created or externally ingested particulate materials that induce erosion and impact damage. Consequently, during operation, the materials of individual components degrade. In addition, the impact of maintenance costs for both civil and military aircraft was estimated at least two to three times greater than initial purchase values, and this trend is expected to increase. As a result, for viable product realisation and maintenance, a spectrum of issues regarding novel processing technologies, innovation of new materials, performance, costs, and environmental impact must constantly be addressed. One of these technologies, namely the cold-gas dynamic-spray process has enabled a broad range of coatings and applications, including many that have not been previously possible or commercially practical, hence its potential for new aerospace applications. Therefore, the purpose of this paper is to summarise the state of the art of this technology alongside its theoretical and experimental studies, and explore how the cold-gas dynamic-spray process could be integrated within a framework that finally could lead to more efficient aircraft maintenance. Based on the paper's qualitative findings supported by authorities, evidence, and logic essentially it is argued that the cold-gas dynamic-spray manufacturing process should not be viewed in isolation, but should be viewed as a component of a broad framework that finally leads to more efficient aerospace operations.Keywords: aerospace, aging aircraft, cold spray, materials
Procedia PDF Downloads 121892 Efficiency of Background Chlorine Residuals against Accidental Microbial Episode in Proto-Type Distribution Network (Rig) Using Central Composite Design (CCD)
Authors: Sajida Rasheed, Imran Hashmi, Luiza Campos, Qizhi Zhou, Kim Keu
Abstract:
A quadratic model (p ˂ 0.0001) was developed by using central composite design of 50 experimental runs (42 non-center + 8 center points) to assess efficiency of background chlorine residuals in combating accidental microbial episode in a prototype distribution network (DN) (rig). A known amount of background chlorine residuals were maintained in DN and a required number of bacteria, Escherichia coli K-12 strain were introduced by an injection port in the pipe loop system. Samples were taken at various time intervals at different pipe lengths. Spread plate count was performed to count bacterial number. The model developed was significant. With microbial concentration and time (p ˂ 0.0001), pipe length (p ˂ 0.022), background chlorine residuals (p ˂ 0.07) and time^2 (p ˂ 0.09) as significant factors. The ramp function of variables shows that at the microbial count of 10^6, at 0.76 L/min, and pipe length of 133 meters, a background residual chlorine 0.16 mg/L was enough for complete inactivation of microbial episode in approximately 18 minutes.Keywords: central composite design (CCD), distribution network, Escherichia coli, residual chlorine
Procedia PDF Downloads 462891 Molecular Detection and Characterization of Shiga Toxogenic Escherichia coli Associated with Dairy Product
Authors: Mohamed Al-Hazmi, Abdullah Al-Arfaj, Moussa Ihab
Abstract:
Raw, unpasteurized milk can carry dangerous bacteria such as Salmonella, E. coli, and Listeria, which are responsible for causing numerous foodborne illnesses. The objective of this study was molecular characterization of shiga toxogenic E. coli in raw milk collected from different Egyptian governorates by multiplex PCR. During the period of 25th May to 25th October 2012, a total of 320 bulk-tank milk samples were collected from 10 cow farms located in different Egyptian governorates. Bacteriological examination of milk samples revealed the presence of E. coli organisms in 65 samples (20.3%), serotyping of the E. coli isolates revealed, 35 strains (10.94%) O111, 15 strains (4.69%) O157: H7, 10 strains (3.13%) O128 and 5 strains (1.56%) O119. Multiplex PCR for detection of shiga toxin type 2 and intimin genes revealed positive amplification of 255 bp fragment of shiga toxin type 2 gene and 384 bp fragment of intimin gene from all E. coli serovar O157: H7, while from serovar O111 were 25 (71.43%), 20 (57.14%) and from serovar O128 were 6 (60%), 8 (80%), respectively. The results of multiplex PCR assay are useful for identification of STEC possessing the eaeA and stx2 genes.Keywords: raw milk, E. coli, multiplex PCR, Shiga toxin type 2, intimin gene
Procedia PDF Downloads 306890 Evaluation of Opposite Type Heterologous MAT Genes Transfer in the Filamentous Fungi Neofusicoccum mediterraneum and Verticillium dahliae
Authors: Stavros Palavouzis, Alexandra Triantafyllopoulou, Aliki Tzima, Epaminondas Paplomatas
Abstract:
Mating-type genes are present in most filamentous fungi, even though teleomorphs for all species have not been recorded. Our study tries to explore the effect of different growth conditions on the expression of MAT genes in Neofusicoccum mediterraneum. As such, selected isolates were grown in potato dextrose broth or in water agar supplemented with pine needles under a 12 h photoperiod, as well as in constant darkness. Mycelia and spores were collected at different time points, and RNA extraction was performed, with the extracted product being used for cDNA synthesis. New primers for MAT gene expression were designed while qPCR results are underway. The second part of the study involved the isolation and cloning in a selected pGEM-T vector of the Botryosphaeria dothidea MAT1 1 1 and MAT1 2 1 mating genes, including flanking regions. As a next step, the genes were amplified using newly designed primers with engineered restriction sites. Amplicons were excised and subsequently sub-cloned in appropriate binary vectors. The constructs were afterward inserted into Agrobacterium tumefaciens and utilized for Agrobacterium-mediated transformation (ATMT) of Neofusicoccum mediterraneum. At the same time, the transformation of a Verticillium dahliae tomato race 1 strain (70V) was performed as a control. While the procedure was successful in regards to V. dahliae, transformed strains of N. mediterraneum could not be obtained. At present, a new transformation protocol, which utilizes a combination of protoplast and Agro transformation, is being evaluated.Keywords: anamorph, heterothallism, perithecia, pycnidia, sexual stage
Procedia PDF Downloads 180889 Health Monitoring of Primates in a Conservation Unit in Brazil
Authors: Elisângela de Albuquerque Sobreira Borovoski, Ricardo Willian Borovoski
Abstract:
Microbiological infections acquired by animals pose a risk to public health. In public health, monitoring the health of primates is linked to the risk of transmission of zoonoses through scratches, bites and contact with biological samples. The project was approved by the Ethics Committee on the Use of Animals Protocol No. 170/2019. It was authorized by ICMBio Protocol No. 52117-1. The study was carried out in the period 2019-2022 in the municipality of Anápolis. Iron and galvanized wire traps were used and the animals were anesthetized with 4.4mg/kg zolethyl intramuscularly and saliva was collected through swabs. Fifty-three capuchin monkeys were captured from the Onofre Quinan Environmental Park in Anápolis-Goiás for health monitoring purposes. In the laboratory, the samples were deposited on the agar surface and seeded by exhaustion to obtain isolated colonies. These colonies were analyzed according to morphocolonial characteristics. Morphometric characterization and biochemical tests for bacterial identification were performed. A total of 861 bacterial samples were isolated, nine of which were strict anaerobic bacteria of the genus Peptostreptococcus. Previous and constant knowledge of the prevalence of pathogenic agents in biological samples is essential to be prepared to act in pandemic situations.Keywords: Brazil, microbiology, monkeys, public health
Procedia PDF Downloads 76888 The Use of Antioxidant and Antimicrobial Properties of Plant Extracts for Increased Safety and Sustainability of Dairy Products
Authors: Loreta Serniene, Dalia Sekmokiene, Justina Tomkeviciute, Lina Lauciene, Vaida Andruleviciute, Ingrida Sinkeviciene, Kristina Kondrotiene, Neringa Kasetiene, Mindaugas Malakauskas
Abstract:
One of the most important areas of product development and research in the dairy industry is the product enrichment with active ingredients as well as leading to increased product safety and sustainability. The most expanding field of the active ingredients is the various plants' CO₂ extracts with aromatic, antioxidant and antimicrobial properties. In this study, 15 plant extracts were evaluated based on their antioxidant, antimicrobial properties as well as sensory acceptance indicators for the development of new dairy products. In order to increase the total antioxidant capacity of the milk products, it was important to determine the content of phenolic compounds and antioxidant activity of CO₂ extract. The total phenolic content of fifteen different commercial CO₂ extracts was determined by the Folin-Ciocalteu reagent and expressed as milligrams of the Gallic acid equivalents (GAE) in gram of extract. The antioxidant activities were determined by 2.2′-azinobis-(3-ethylbenzthiazoline)-6-sulfonate (ABTS) methods. The study revealed that the antioxidant activities of investigated CO₂ extract vary from 4.478-62.035 µmole Trolox/g, while the total phenolic content was in the range of 2.021-38.906 mg GAE/g of extract. For the example, the estimated antioxidant activity of Chinese cinnamon (Cinammonum aromaticum) CO₂ extract was 62.023 ± 0.15 µmole Trolox/g and the total flavonoid content reached 17.962 ± 0.35 mg GAE/g. These two parameters suggest that cinnamon could be a promising supplement for the development of new cheese. The inhibitory effects of these essential oils were tested by using agar disc diffusion method against pathogenic bacteria, most commonly found in dairy products. The obtained results showed that essential oil of lemon myrtle (Backhousia citriodora) and cinnamon (Cinnamomum cassia) has antimicrobial activity against E. coli, S. aureus, B. cereus, P. florescens, L. monocytogenes, Br. thermosphacta, P. aeruginosa and S. typhimurium with the diameter of inhibition zones variation from 10 to 52 mm. The sensory taste acceptability of plant extracts in combination with a dairy product was evaluated by a group of sensory evaluation experts (31 individuals) by the criteria of overall taste acceptability in the scale of 0 (not acceptable) to 10 (very acceptable). Each of the tested samples included 200g grams of natural unsweetened greek yogurt without additives and 1 drop of single plant extract (essential oil). The highest average of overall taste acceptability was defined for the samples with essential oils of orange (Citrus sinensis) - average score 6.67, lemon myrtle (Backhousia citriodora) – 6.62, elderberry flower (Sambucus nigra flos.) – 6.61, lemon (Citrus limon) – 5.75 and cinnamon (Cinnamomum cassia) – 5.41, respectively. The results of this study indicate plant extracts of Cinnamomum cassia and Backhousia citriodora as a promising additive not only to increase the total antioxidant capacity of the milk products and as alternative antibacterial agent to combat pathogenic bacteria commonly found in dairy products but also as a desirable flavour for the taste pallet of the consumers with expressed need for safe, sustainable and innovative dairy products. Acknowledgment: This research was funded by the European Regional Development Fund according to the supported activity 'Research Projects Implemented by World-class Researcher Groups' under Measure No. 01.2.2-LMT-K-718.Keywords: antioxidant properties, antimicrobial properties, cinnamon, CO₂ plant extracts, dairy products, essential oils, lemon myrtle
Procedia PDF Downloads 204887 Analyzing Healthy Eating Among Adolescent Teens Using the Socioecological Model
Authors: Kaavya Chandrasekar
Abstract:
Healthy eating is essential to maintain good health and stable mental status regardless of age. WHO describes that a healthy diet consists of incorporating more fruits and vegetables and reducing the consumption of sugary and salty foods into a regularly scheduled healthy diet. Although this attitude is rather uncommon among all age groups, it is notably uncommon among the teens being a very vulnerable state in a man’s life. Faulty dietary habits, in the long run, interfere with health, leading to obesity, cardiovascular diseases, and mental instability. This study collates a discussion on the barriers prevailing among adolescents, to inculcate healthy eating practices by means of the socioecological model. The studies consisted of teens aged 13 to 19 years from schools and colleges of both sexes. The socio-ecological model emphasizes the interplay and interconnectedness of elements at all levels of health behavior, acknowledging that the majority of public health issues are just too complicated to be solved from a single-level perspective. As a result, it necessitates that people are not considered in isolation from bigger social groups. According to the studies retrieved from ten articles studies conducted globally, more than five articles suggest that socioeconomic class, lack of adult supervision and easy access to fast food stores and schools affect their decision of healthy eating. Awareness via personalized intervention has been tried and found successful. Future research is still needed to address various dimensions of the issue.Keywords: socio ecological model, healthy eating, adolescents, fast food consumption, interventions.
Procedia PDF Downloads 25886 Synergistic Interactions between Secondary Metabolites in Rosmarinus officinalis L.
Authors: Ruta Mickiene, Audrius Maruska, Ona Ragazinskiene
Abstract:
This research focuses on phytochemistry and antimicrobial activities of compounds isolated and identified from species Rosmarinus officinalis L. This is a study of synergistic effects between phenolic fraction and essential oils. The antimicrobial activity of extracts from Rosmarinus officinalis L. originated from the sector of medicinal plants, Kaunas botanical garden of Vytautas Magnus University Lithuania, were tested by the method of series dilutions, against different bacteria species. Investigated microorganisms were Escherichia coli, Proteus vulgaris and Staphylococcus aureus with and without antibiotic resistances originating from livestock. The antimicrobial activities of extracts were described by determination of the Minimal Inhibitory Concentration (MIC). Preliminary results show that the MIC range between 9.0 % and 12.0 % for the different Rosmarinus officinalis L. extracts and bacterial species. The total amounts of phenolic compounds and total amounts of flavonoids were tested in the methanolic extracts of the plants. The chemical composition for essential oils analysed by GC/MS. Predominant components were alpha pinene (20%), camphor (10%), 1.8‐cineole (5%), phellandrene (6%), camphene (5%), beta pinene (4%), bornylacetate (4%), limonene (2%), borneol (3%), alpha terpineol (3%), cymene (2%), caryophyllene (15%), verbenone (7%), alpha terpinene (3%), eucalyptol (11%).Keywords: antimicrobial activity, essential oil, Rosmarinus officinalis L., escherichia coli
Procedia PDF Downloads 384885 Preparation and Characterization of Maltodextrin Microcapsules Containing Walnut Green Husk Extract
Authors: Fatemeh Cheraghali, Saeedeh Shojaee-Aliabadi, Seyede Marzieh Hosseini, Leila Mirmoghtadaie
Abstract:
In recent years, the field of natural antimicrobial and antioxidant compounds is one of the main research topics in the food industry. Application of agricultural residues is mainly cheap, and available resources are receiving increased attention. Walnut green husk is one of the agricultural residues that is considered as natural compounds with biological properties because of phenolic compounds. In this study, maltodextrin 10% was used for microencapsulation of walnut green husk extract. At first, the extract was examined to consider extraction yield, total phenolic compounds, and antioxidant activation. The results showed the extraction yield of 81.43%, total phenolic compounds of 3997 [mg GAE/100 g], antioxidant activity [DPPH] of 84.85% for walnut green husk extract. Antioxidant activity is about 75%-81% and by DPPH. At the next stage, microencapsulation was done by spry-drying method. The microencapsulation efficiency was 72%-79%. The results of SEM tests confirmed this microencapsulation process. In addition, microencapsulated and free extract was more effective on gram-positive bacteria’s rather than the gram-negative ones. According to the study, walnut green husk can be used as a cheap antioxidant and antimicrobial compounds due to sufficient value of phenolic compounds.Keywords: biopolymer, microencapsulation, spray-drying, walnut green husk
Procedia PDF Downloads 161884 Effect of Citric Acid and Clove on Cured Smoked Meat: A Traditional Meat Product
Authors: Esther Eduzor, Charles A. Negbenebor, Helen O. Agu
Abstract:
Smoking of meat enhances the taste and look of meat, it also increases its longevity, and helps preserve the meat by slowing down the spoilage of fat and growth of bacteria. The Lean meat from the forequarter of beef carcass was obtained from the Maiduguri abattoir. The meat was cut into four portions with weight ranging from 525-545 g. The meat was cut into bits measuring about 8 cm in length, 3.5 cm in thickness and weighed 64.5 g. Meat samples were washed, cured with various concentration of sodium chloride, sodium nitrate, citric acid and clove for 30 min, drained and smoked in a smoking kiln at a temperature range of 55-600°C, for 8 hr a day for 3 days. The products were stored at ambient temperature and evaluated microbiologically and organoleptically. In terms of processing and storage there were increases in pH, free fatty acid content, a decrease in water holding capacity and microbial count of the cured smoked meat. The panelists rated control samples significantly (p < 0.05) higher in terms of colour, texture, taste and overall acceptability. The following organisms were isolated and identified during storage: Bacillus specie, Bacillus subtilis, streptococcus, Pseudomonas, Aspergillus niger, Candida and Penicillium specie. The study forms a basis for new product development for meat industry.Keywords: citric acid, cloves, smoked meat, bioengineering
Procedia PDF Downloads 445883 Antibacterial Activity of Trans-Cinnamaldehyde and Geraniol and Their Potential as Ingredients of Biocidal Polymers
Authors: Daria Olkiewicz, Maciej Walczak
Abstract:
In this paper, the biocidal effects of trans-cinnamaldehyde (a main component of cinnamon oil) and geraniol (a constituent of Pelargonium graveolens essential oil) are presented. The activity of the combination of trans-cinnamaldehyde and geraniol was tested against 3 bacterial strains: Staphylococcus aureus ATCC6538 (Gramm+), Escherichia coli ATCC8739 (Gramm-, Lac+) and Pseudomonas aeruginosa KKP 991(Gramm-, Lac-). The biocidal activity of trans-cinnamaldehyde-geraniol mixture against bacteria mentioned above was evaluated by disk-diffusion method. The model strains were exposed on 1, 2.5, 5 and 10 mg of trans-cinnamaldehyde-geraniol mixture per disk, and all strains were susceptible to this combination of plant compounds. For all microorganisms, also Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were estimated. For Staphylococcus aureus MIC was 0.0625 mg/ml of the trans-cinnamaldehyde and geraniol mixture, and MBC was 1.25 mg/ml; For Escherichia coli MIC=0.5 mg/ml, MBC=1 mg/ml, and finally Pseudomonas aeruginosa was inhibited in 0.5 mg/ml, and minimal biocidal concentration of tested mixture for it was 1.25 mg/ml. There are also reports about the synergistic working of trans-cinnamaldehyde and geraniol against microorganisms and the antimicrobial activity of polymers enriched with trans-cinnamaldehyde or geraniol, therefore the successful development and introduction to the today life of biocidal polymer enriched with trans-cinnamaldehyde and geraniol are possible.Keywords: antibacterial activity, biocidal polymers, geraniol, trans-cinnamaldehyde
Procedia PDF Downloads 177882 Essential Oil Composition and Antimicrobial Activity of Rosmarinus officinalis L. Grown in Algeria (Djelfa)
Authors: Samah Lakehal, A. Meliani, F. Z. Benrebiha, C. Chaouia
Abstract:
In the last few years, due to the misuse of antibiotics and an increasing incidence of immunodeficiency-related diseases, the development of microbial drug resistance has become more and more of a pressing problem. Recently, natural products from medicinal plants represent a fertile ground for the development of novel antibacterial agents. Plants essential oils have come more into the focus of phytomedicine. The present study describes antimicrobial activity of Rosmarinus officinalis L. essential oil known medicinally for its powerful antibacterial properties. The essential oil of rosemary obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (Djelfa city of south Algeria) was investigated by GC-MS. The essential oil yield of the study was 1.4 %. The major components were found to be camphor, camphene, 1,8-cineole. The essential oil has been tested for antimicrobial activity against eight bacteria (Gram-negative and Gram-positive), and three fungi including Candida albicans. Inhibition of growth was tested by the agar diffusion method based on the determination of the diameter of inhibition. The oil was found to have significant antibacterial activity and therefore can be used as a natural antimicrobial agent for the treatment of several infectious diseases caused by those germs, which have developed resistance to antibiotics.Keywords: antimicrobial activity, Rosmarinus officinalis L., essential oils, GC/MS, camphor
Procedia PDF Downloads 391881 A Furaneol-Containing Glass-Ionomer Cement for Enhanced Antibacterial Activity
Authors: Dong Xie, Yuling Xu, Leah Howard
Abstract:
Secondary caries is found to be one of the main reasons to the restoration failure of dental restoratives. To prevent secondary caries formation, dental restoratives ought to be made antibacterial. In this study, a natural fruit component furaneol was tethered onto polyacid, the formed polyacid was used to formulate the light-curable glass-ionomer cements, and then the effect of this new antibacterial compound on compressive strength (CS) and antibacterial activity of the formed cement was evaluated. Fuji II LC glass powders were used as fillers. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the formed cement. The experimental cement showed a significant antibacterial activity, accompanying with an initial CS reduction. Increasing the compound loading significantly decreased the S. mutans viability from 5 to 81% and also reduced the initial CS of the formed cements from 4 to 58%. The cement loading with 7% antibacterial polymer showed 168 MPa, 7.8 GPa, 243 MPa, 46 MPa, and 57 MPa in yield strength, modulus, CS, diametral tensile strength and flexural strength, respectively, as compared to 141, 6.9, 236, 42 and 53 for Fuji II LC. The cement also showed an antibacterial function to other bacteria. No human saliva effect was noticed. It is concluded that the experimental cement may potentially be developed to a permanent antibacterial cement.Keywords: antibacterial, dental materials, strength, cell viability
Procedia PDF Downloads 317880 Effect of Phosphate and Zinc Biofertilizers on Seed Yield and Molar Ratio of Phytic Acid to Zinc in Two Cultivars of Bean (Phaseolus vulgaris L.)
Authors: M. Mohammadi
Abstract:
In order to evaluate the effect of phosphate and Zn bio-fertilizers on the yield, phytic acid (PA), Zn concentration and PA/Zn molar ratio in bean, a field experiment was carried out for two years. The treatments included two cultivars of bean (Talash and Sadri), four levels of P (P0, P1: 100 kg ha-1 triple super phosphate (TSP), P2: 50 kg ha-1 TSP + phosphate bio-fertilizer, P3: phosphate bio-fertilizer), three levels of Zn (Zn0, Zn1: 50 kg ha-1 ZnSO4, Zn2: Zn bio-fertilizer). Phosphate bio-fertilizer consisted of inoculum of mycorrhizal fungus and Azotobacter and Zn bio-fertilizer consisted of Pseudomonas bacteria. The results revealed that there was significant difference between yield and Zn concentration between years. The effect of cultivar was significant on studied parameters. The lowest content of PA and PA/Zn were obtained from Talash. P treatment caused to significant difference on parameters in which P2 caused to increase yield, P and Zn concentration, and decrease PA and PA/Zn by 21.8%, 38.2%, 33.4%, 17.4% and 38.6% respectively. Zn treatment caused to significant difference on studied parameters. The maximum number of parameters were obtained from Zn1 and Zn2. The higher Zn concentration led to lower content of PA and PA/Zn. Using of P and Zn bio–fertilizers were caused to increasing nutrient uptake, improving growth condition and reducing PA and PA/Zn molar ratio.Keywords: mycorrhizae, phosphorus, pseudomonas, zinc
Procedia PDF Downloads 262879 Properties and Antimicrobial Activity of Fish Protein Isolate/Fish Skin Gelatin Film Containing Basil Leaf Essential Oil and Zinc Oxide Nanoparticles
Authors: Yasir Ali Arfat
Abstract:
Composite films based on fish protein isolate (FPI) and fish skin gelatin (FSG) blend incorporated with 50 and 100% (w/w, protein) basil leaf essential oil (BEO) in the absence and presence of 3% (w/w, protein) ZnO nanoparticles (ZnONP) were prepared and characterised. Tensile strength (TS) decreased, whilst elongation at break (EAB) increased as BEO level increased (p < 0.05). However, ZnONP addition resulted in higher TS but lower EAB (p < 0.05). The lowest water vapour permeability (WVP) was observed for the film incorporated with 100% BEO and 3% ZnONP (p < 0.05). BEO and ZnONP incorporation decreased transparency of FPI/FSG films (p < 0.05). FTIR spectra indicated that films added with BEO exhibited higher hydrophobicity. Both BEO and ZnONP had a marked impact on thermal stability of the films. Microstructural study revealed that presence of ZnONP prevented bilayer formation of film containing 100% BEO. FPI/FSG films incorporated with 100% BEO, especially in combination with ZnONP, exhibited strong antibacterial activity against food pathogenic and spoilage bacteria and thus could be used as an active food packaging material to ensure safety and to extend the shelf-life of packaged foods.Keywords: bionanocomposite, fish protein isolate, fish skin gelatin, basil essential oil, ZnO nanoparticles, antimicrobial packaging
Procedia PDF Downloads 471878 The Nutritive Value of Fermented Sago Pith (Metroxylon sago Rottb) Enriched with Micro Nutrients for Poultry Feed
Authors: Wizna, Helmi Muis, Hafil Abbas
Abstract:
An experiment was conducted to improve the nutrient value of sago pith (Metroxylon sago Rottb) supplemented with Zn, Sulfur and urea through fermentation by using cellulolytic bacteria (Bacillus amyloliquefaciens) as inoculums. The experiment was determination of the optimum dose combination (dosage of Zn, S and urea) for sago pith fermentation based on nutrient quality and quantity of these fermented products. The study was conducted in experimental method, using the completely randomized design in factorial with 3 treatments consist of: factor A (Dose of urea: A1 = 2.0%, A2 = 3.0%), factor B (Dose of S: B1 = 0.2%, B2 = 0.4%) and factor C (Dose of Zn: C1 = 0.0025%, C2 = 0.005%). Results of study showed that optimum condition for fermentation process of sago pith with B. amyloliquefaciens caused a change of nutrient content was obtained at urea (3%), S (0,2%), and Zn (0,0025%). This fermentation process was able to increase amino acid average, reduce crude fiber content by 67% and increase crude protein by 433%, which made the nutritional value of the product based on dry matter was 18.22% crude protein, 12.42% crude fiber, 2525 Kcal/kg metabolic energy and 65.73% nitrogen retention.Keywords: fermentation, sago pith, sulfur, Zn, urea, Bacillus amyloliquefaciens
Procedia PDF Downloads 511877 Identification, Isolation and Characterization of Unknown Degradation Products of Cefprozil Monohydrate by HPTLC
Authors: Vandana T. Gawande, Kailash G. Bothara, Chandani O. Satija
Abstract:
The present research work was aimed to determine stability of cefprozil monohydrate (CEFZ) as per various stress degradation conditions recommended by International Conference on Harmonization (ICH) guideline Q1A (R2). Forced degradation studies were carried out for hydrolytic, oxidative, photolytic and thermal stress conditions. The drug was found susceptible for degradation under all stress conditions. Separation was carried out by using High Performance Thin Layer Chromatographic System (HPTLC). Aluminum plates pre-coated with silica gel 60F254 were used as the stationary phase. The mobile phase consisted of ethyl acetate: acetone: methanol: water: glacial acetic acid (7.5:2.5:2.5:1.5:0.5v/v). Densitometric analysis was carried out at 280 nm. The system was found to give compact spot for cefprozil monohydrate (0.45 Rf). The linear regression analysis data showed good linear relationship in the concentration range 200-5.000 ng/band for cefprozil monohydrate. Percent recovery for the drug was found to be in the range of 98.78-101.24. Method was found to be reproducible with % relative standard deviation (%RSD) for intra- and inter-day precision to be < 1.5% over the said concentration range. The method was validated for precision, accuracy, specificity and robustness. The method has been successfully applied in the analysis of drug in tablet dosage form. Three unknown degradation products formed under various stress conditions were isolated by preparative HPTLC and characterized by mass spectroscopic studies.Keywords: cefprozil monohydrate, degradation products, HPTLC, stress study, stability indicating method
Procedia PDF Downloads 299876 Application of Box-Behnken Response Surface Design for Optimization of Essential Oil Based Disinfectant on Mixed Species Biofilm
Authors: Anita Vidacs, Robert Rajko, Csaba Vagvolgyi, Judit Krisch
Abstract:
With the optimization of a new disinfectant the number of tests could be decreased and the cost of processing too. Good sanitizers are eco-friendly and allow no resistance evolvement of bacteria. The essential oils (EOs) are natural antimicrobials, and most of them have the Generally Recognized As Safe (GRAS) status. In our study, the effect of the EOs cinnamon, marjoram, and thyme was investigated against mixed species bacterial biofilms of Escherichia coli, Listeria monocytogenes, Pseudomonas putida, and Staphylococcus aureus. The optimal concentration of EOs, disinfection time and level of pH were evaluated with the aid of Response Surface Box-Behnken Design (RSD) on 1 day and 7 days old biofilms on metal, plastic, and wood surfaces. The variable factors were in the range of 1-3 times of minimum bactericide concentration (MBC); 10-110 minutes acting time and 4.5- 7.5 pH. The optimized EO disinfectant was compared to industrial used chemicals (HC-DPE, Hypo). The natural based disinfectants were applicable; the acting time was below 30 minutes. EOs were able to eliminate the biofilm from the used surfaces except from wood. The disinfection effect of the EO based natural solutions was in most cases equivalent or better compared to chemical sanitizers used in food industry.Keywords: biofilm, Box-Behnken design, disinfectant, essential oil
Procedia PDF Downloads 220875 Media Manipulations and the Culture of Beneficial Endophytic Fungi in the Leaves and Stem Bark of Grewia lasiocarpa E. Mey. Ex Harv
Authors: Akwu A. Nneka, Naidoo, Yougasphree
Abstract:
A significantly high number of microbes exist in higher plants; these microbes include bacteria, fungi, and actinomycetes. There are reports on the benefits of endophytic fungi and their products of metabolism to the host plant and man, consequently, it is expedient to explore the changes that could arise as a result of manipulating their growth media. Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is an indigenous Southern African plant, that belongs to a genus with known medicinal properties. Three media were used to culture the endophytic fungi viz., Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), and Bacteriological Agar (BA) were used singly, and supplemented with three dilutions of the leaves and stem bark extracts. The manipulated growth media composition had a significant effect on the diversity of the isolated fungal populations. Several endophytic fungi were isolated; their distribution and diversity revealed a significant relatedness with the manipulated media. The media supplemented with the plant extracts was observed to give a significant increase in the growth rate and yield of the endophytes. To the best of our knowledge, this is the first study describing the endophytic fungi present in the leaves and stem bark of G. lasiocarpa E. Mey. ex Harv.Keywords: Grewia lasiocarpa, plant-based extracts, endophytic fungi, Malvaceae
Procedia PDF Downloads 155874 The Photocatalytic Approach for the Conversion of Polluted Seawater CO₂ into Renewable Source of Energy
Authors: Yasar N. Kavil, Yasser A. Shaban, Radwan K. Al Farawati, Mohamed I. Orif, Shahed U. M. Khanc
Abstract:
Photocatalytic way of reduction of CO₂ in polluted seawater into chemical fuel, methanol, was successfully gained over Cu/C-co-doped TiO₂ nanoparticles under UV and natural sunlight. A homemade stirred batch annular reactor was used to carry out the photocatalytic reduction experiments. Photocatalysts with various Cu loadings (0, 0.5, 1, 3, 5 and 7 wt.%) were synthesized by the sol-gel procedure and were characterized by XRD, SEM, UV–Vis, FTIR, and XPS. The photocatalytic production of methanol was promoted by the co-doping with C and Cu into TiO₂. This improvement was attributed to the modification of bandgap energy and the hindrance of the charges recombination. The polluted seawater showing the yield depended on its background hydrographic parameters. We assessed two types of polluted seawater system, the observed yield was 2910 and 990 µmol g⁻¹ after 5 h of illumination under UV and natural sunlight respectively in system 1 and the corresponding yield in system 2 was 2250 and 910 µmol g⁻¹ after 5 h of illumination. The production of methanol in the case of oxygen-depleted water was low, this is mainly attributed to the competition of methanogenic bacteria over methanol production. The results indicated that the methanol yield produced by Cu-C/TiO₂ was much higher than those of carbon-modified titanium oxide (C/TiO₂) and Degussa (P25-TiO₂). Under the current experimental condition, the optimum loading was achieved by the doping of 3 wt % of Cu. The highest methanol yield was obtained over 1 g L-1 of 3wt% Cu/C-TiO₂.Keywords: CO₂ photoreduction, copper, Cu/C-co-doped TiO₂, methanol, seawater
Procedia PDF Downloads 278873 A Case Study on the Census of Technological Capacities in Health Care in Rural Sanitary Institutions in South Cameroon
Authors: Doriane Micaela Andeme Bikoro, Samuel Fosso Wamba, Jean Robert Kala Kamdjoug
Abstract:
Currently one of the leading fields in the market of technological innovation is digital health. In developed countries, this booming innovation is experiencing an exponential speed. We understand that in developed countries, e-health could also revolutionize the practice of medicine and therefore fill the many failures observed in medical care. Everything leads to believe that future technology is oriented towards the medical sector. The aim of this work is to explore at the same time the technological resources and the potential of health care based on new technologies; it is a case study in a rural area of Southern Cameroon. Among other things, we will make a census of the shortcomings and problems encountered, and we will propose various appropriate solutions. The work methodology used here is essentially qualitative. We used two qualitative data collection techniques, direct observation, and interviews. In fact, we spent two weeks in the field observing and conducting some semi-directive interviews with some of those responsible for these health structures. This study was conducted in three health facilities in the south of the country; including two health centers and a rural hospital. Many technological failures have been identified in the day-to-day management of these health facilities and especially in the administration of health care to patients. We note major problems such as the digital divide, the lack of qualified personnel, the state of isolation of this area. This is why various proposals are made to improve the health sector in Cameroon both technologically and medically.Keywords: Cameroon, capacities, census, digital health, qualitative method, rural area
Procedia PDF Downloads 144872 RNA-Seq Based Transcriptomic Analysis of Wheat Cultivars for Unveiling of Genomic Variations and Isolation of Drought Tolerant Genes for Genome Editing
Authors: Ghulam Muhammad Ali
Abstract:
Unveiling of genes involved in drought and root architecture using transcriptomic analyses remained fragmented for further improvement of wheat through genome editing. The purpose of this research endeavor was to unveil the variations in different genes implicated in drought tolerance and root architecture in wheat through RNA-seq data analysis. In this study seedlings of 8 days old, 6 cultivars of wheat namely, Batis, Blue Silver, Local White, UZ888, Chakwal 50 and Synthetic wheat S22 were subjected to transcriptomic analysis for root and shoot genes. Total of 12 RNA samples was sequenced by Illumina. Using updated wheat transcripts from Ensembl and IWGC references with 54,175 gene models, we found that 49,621 out of 54,175 (91.5%) genes are expressed at an RPKM of 0.1 or more (in at least 1 sample). The number of genes expressed was higher in Local White than Batis. Differentially expressed genes (DEG) were higher in Chakwal 50. Expression-based clustering indicated conserved function of DRO1and RPK1 between Arabidopsis and wheat. Dendrogram showed that Local White is sister to Chakwal 50 while Batis is closely related to Blue Silver. This study flaunts transcriptomic sequence variations in different cultivars that showed mutations in genes associated with drought that may directly contribute to drought tolerance. DRO1 and RPK1 genes were fetched/isolated for genome editing. These genes are being edited in wheat through CRISPR-Cas9 for yield enhancement.Keywords: transcriptomic, wheat, genome editing, drought, CRISPR-Cas9, yield enhancement
Procedia PDF Downloads 147871 Preparation and in vitro Bactericidal and Fungicidal Efficiency of NanoSilver/Methylcellulose Hydrogel
Authors: A. Panacek, M. Kilianova, R. Prucek, V. Husickova, R. Vecerova, M. Kolar, L. Kvitek, R. Zboril
Abstract:
In this work we describe the preparation of NanoSilver/methylcellulose hydrogel containing silver nanoparticles (NPs) for topical bactericidal applications. Highly concentrated dispersion of silver NPs as high as of 5g/L of silver with diameter of 10nm was prepared by reduction of AgNO3 via strong reducing agent NaBH4. Silver NPs were stabilized by addition of sodium polyacrylate in order to prevent their aggregation at such high concentration. This way synthesized silver NPs were subsequently incorporated into methylcellulose suspension at elevated temperature resulting in formation of NanoSilver/methylcellulose hydrogel when temperature cooled down to laboratory conditions. In vitro antibacterial activity assay proved high bactericidal and fungicidal efficiency of silver NPs alone in the form of dispersion as well as in the form of hydrogel against broad spectrum of bacteria and yeasts including highly multiresistant strains such as methicillin-resistant Staphylococcus aureus. A very low concentrations of silver as low as 0.84mg/L Ag in as-prepared dispersion gave antibacterial performance. NanoSilver/methylcellulose hydrogel showed antibacterial action at the lowest used silver concentration equal to 25mg/L. Such prepared NanoSilver/methylcellulose hydrogel represent promising topical antimicrobial formulation for treatment of burns and wounds.Keywords: antimicrobial, burn, hydrogel, silver NPs
Procedia PDF Downloads 451870 Analysis to the Characterization of Self-Esteem of Students in Eulogio 'Amang' Rodriguez Institute of Science and Technology: A Foundation of Conceptualizing Substantial Plan of Action
Authors: Eriberto R. Astorga Jr., Herbert D. Vertucio, Evelyn M. Polison
Abstract:
This study was conducted in order to determine the analysis with regards to the Characterization of Self-Esteem of EARIST Students according to their origin of self-esteem and low self-esteem as well as its causes. The respondents of this study are three thousand three hundred twenty three (3,323) randomly selected students from eight colleges of EARIST such as Arts and Sciences, Education, Public Administration and Criminology, Business Administration, Hospitality Management, Architecture and Fine Arts, Engineering, and Industrial Technology. A survey was conducted by using a validated questionnaire for information gathering about respondents profile and different factors relating to self-esteem of students such as self-origin, familial and social relationship, financial situation and education. Frequency, percentage, ranking and standards deviation, standard t-test and ANOVA were applied to investigate the differences of the answers of the respondents to the origin of their self-esteem and the reasons for low self-esteem. The results revealed that there are no significant differences in the origin of their self-esteem and the reasons of low esteem as to the eight group of respondent’s. Moreover, most causes of low esteem are caused by hearing a comment or experiencing an incident that has a negative impact student mentally and emotionally, poor health, being bullied, lack of support from family, friends, and job loss, experiencing verbal and sexual abuse and are in a violent relationship, feelings of isolation, divorce, dysfunctional family, death and lack of achievement at work and at school, trying to conform to stereotypes and prove our independence from our parents.Keywords: characterization, plan of action, profile, self-esteem
Procedia PDF Downloads 175869 Comparison of Rumen Microbial Analysis Pipelines Based on 16s rRNA Gene Sequencing
Authors: Xiaoxing Ye
Abstract:
To investigate complex rumen microbial communities, 16S ribosomal RNA (rRNA) sequencing is widely used. Here, we evaluated the impact of bioinformatics pipelines on the observation of OTUs and taxonomic classification of 750 cattle rumen microbial samples by comparing three commonly used pipelines (LotuS, UPARSE, and QIIME) with Usearch. In LotuS-based analyses, 189 archaeal and 3894 bacterial OTUs were observed. The observed OTUs for the Usearch analysis were significantly larger than the LotuS results. We discovered 1495 OTUs for archaea and 92665 OTUs for bacteria using Usearch analysis. In addition, taxonomic assignments were made for the rumen microbial samples. All pipelines had consistent taxonomic annotations from the phylum to the genus level. A difference in relative abundance was calculated for all microbial levels, including Bacteroidetes (QIIME: 72.2%, Usearch: 74.09%), Firmicutes (QIIME: 18.3%, Usearch: 20.20%) for the bacterial phylum, Methanobacteriales (QIIME: 64.2%, Usearch: 45.7%) for the archaeal class, Methanobacteriaceae (QIIME: 35%, Usearch: 45.7%) and Methanomassiliicoccaceae (QIIME: 35%, Usearch: 31.13%) for archaeal family. However, the most prevalent archaeal class varied between these two annotation pipelines. The Thermoplasmata was the top class according to the QIIME annotation, whereas Methanobacteria was the top class according to Usearch.Keywords: cattle rumen, rumen microbial, 16S rRNA gene sequencing, bioinformatics pipeline
Procedia PDF Downloads 88868 Isolation, Purification and Characterisation of Non-Digestible Oligosaccharides Derived from Extracellular Polysaccharide of Antarctic Fungus Thelebolus Sp. IITKGP-BT12
Authors: Abinaya Balasubramanian, Satyabrata Ghosh, Satyahari Dey
Abstract:
Non-Digestible Oligosaccharides(NDOs) are low molecular weight carbohydrates with degree of polymerization (DP) 3-20, that are delivered intact to the large intestine. NDOs are gaining attention as effective prebiotic molecules that facilitate prevention and treatment of several chronic diseases. Recently, NDOs are being obtained by cleaving complex polysaccharides as it results in high yield and also as the former tend to display greater bioactivity. Thelebolus sp. IITKGP BT-12, a recently identified psychrophilic, Ascomycetes fungus has been reported to produce a bioactive extracellular polysaccharide(EPS). The EPS has been proved to possess strong prebiotic activity and anti- proliferative effects. The current study is an attempt to identify and optimise the most suitable method for hydrolysis of the above mentioned novel EPS into NDOs, and further purify and characterise the same. Among physical, chemical and enzymatic methods, enzymatic hydrolysis was identified as the best method and the optimum hydrolysis conditions obtained using response surface methodology were: reaction time of 24h, β-(1,3) endo-glucanase concentration of 0.53U and substrate concentration of 10 mg/ml. The NDOs were purified using gel filtration chromatography and their molecular weights were determined using MALDI-TOF. The major fraction was found to have a DP of 7,8. The monomeric units of the NDOs were confirmed to be glucose using TLC and GCMS-MS analysis. The obtained oligosaccharides proved to be non-digestible when subjected to gastric acidity, salivary and pancreatic amylases and hence could serve as efficient prebiotics.Keywords: characterisation, enzymatic hydrolysis, non-digestible oligosaccharides, response surface methodology
Procedia PDF Downloads 129867 Biodegradation Study of Diethyl Phthalate Using Bacteria Isolated from Plastic Industry Wastewater Discharge Site
Authors: Sangram Shamrao Patil, Hara Mohan Jena
Abstract:
Phthalates are among the most common organic pollutant since they have become widespread in the environment and found in sediments, natural waters, soils, plants, landfill leachates, biota including human tissue and aquatic organisms. Diethyl phthalate (DEP) is a low molecular weight phthalate which has wide applications as plasticizer and become a major cause of environmental pollution. Environmental protection agency (EPA) listed DEP as priority pollutant because of its toxicity and they recommended human health ambient water quality criterion for diethyl phthalate (DEP) as 4 mg/l. Therefore, wastes containing phthalates require proper treatment before being discharged into the environment. Biodegradation is attractive and efficient treatment method as it is cost effective and produces non-toxic end products. In the present study, a DEP degrading aerobic bacterium was isolated from soil contaminated with plastic industry wastewater. Morphological and biochemical characteristics of isolate were performed. 16S rRNA sequencing and phylogenetic analysis of isolate was carried out and it was identified as Empedobacter brevis. Isolate has been found to tolerate up to 1650 mg/l of DEP. This study will be significant for exploring an application of microbes for remediation of phthalates and development of a suitable bioreactor.Keywords: diethyl phthalate, plasticizer, pollutant, biodegradation
Procedia PDF Downloads 271