Search results for: S. mutans viability
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 713

Search results for: S. mutans viability

623 Preservation of Coconut Toddy Sediments as a Leavening Agent for Bakery Products

Authors: B. R. Madushan, S. B. Navaratne, I. Wickramasinge

Abstract:

Toddy sediment (TS) was cultured in a PDA medium to determine initial yeast load, and also it was undergone sun, shade, solar, dehumidified cold air (DCA) and hot air oven (at 400, 500 and 60oC) drying with a view to preserve viability of yeast. Thereafter, this study was conducted according to two factor factorial design in order to determine best preservation method. Therein the dried TS from the best drying method was taken and divided into two portions. One portion was mixed with 3: 7 ratio of TS: rice flour and the mixture was divided in to two again. While one portion was kept under in house condition the other was in a refrigerator. Same procedure was followed to the rest portion of TS too but it was at the same ratio of corn flour. All treatments were vacuum packed in triple laminate pouches and the best preservation method was determined in terms of leavening index (LI). The TS obtained from the best preservation method was used to make foods (bread and hopper) and organoleptic properties of it were evaluated against same of ordinary foods using sensory panel with a five point hedonic scale. Results revealed that yeast load or fresh TS was 58×106 CFU/g. The best drying method in preserving viability of yeast was DCA because LI of this treatment (96%) is higher than that of other three treatments. Organoleptic properties of foods prepared from best preservation method are as same as ordinary foods according to Duo trio test.

Keywords: biological leavening agent, coconut toddy, fermentation, yeast

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622 Performance of a Lytic Bacteriophage Cocktail against Pseudomonas aeruginosa in Conditions That Simulate the Cystic Fibrosis Lung Environment

Authors: Isaac Martin, Abigail Lark, Sandra Morales, Eric W. Alton, Jane C. Davies

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Objectives: The cystic fibrosis (CF) lung is a unique microbiological niche, wherein harmful bacteria persist for many years despite antibiotic therapy. Pseudomonas aeruginosa (Pa), the major culprit leading to lung decline and increased mortality, thrives in the lungs of patients with CF due to several factors that have been linked with poor antibiotic performance. Our group is investigating alternative therapies including bacteriophage cocktails with which we have previously demonstrated efficacy against planktonic organisms. In this study, we explored the effects of a 4-phage cocktail on Pa grown in two different conditions, intended to mirror the CF lung: a) alongside standard antibiotic treatment in pre-formed biofilms (structures formed by Pa-secreted exopolysaccharides which provide both physical and cell division barriers to antimicrobials and host defenses and b) in an acidic environment postulated to be present in the CF airway due both to the primary defect in bicarbonate secretion and secondary effects of inflammation. Methods: 16 Pa strains from CF patients at the Royal Brompton Hospital were selected based on sensitivity to a) ceftazidime/ tobramycin and b) the phage cocktail in a conventional plaque assay. To assess efficacy of phage in biofilms, 96 well plates with Pa (5x10⁷ CFU/ ml) were incubated in static conditions, allowing adherent bacterial colonies to form for 24 hr. Ceftazidime and tobramycin (both at 2 × MIC) were added, +/- bacteriophage (4x10⁸ PFU/mL) for a further 24 hr. Cell viability and biomass were estimated using fluorescent resazurin and crystal violet assays, respectively. To evaluate the effect of pH, strains were grown planktonically in shaking 96 well plates at pH 6.0, 6.6, 7.0 and 7.5 with tobramycin or phage, at varying concentrations. Cell viability was quantified by fluorescent resazurin assay. Results: For the biofilm assay, treatment groups were compared with untreated controls and expressed as percent reduction in cell viability and biomass. Addition of the 4-phage cocktail resulted in a 1.3-fold reduction in cell viability and 1.7-fold reduction in biomass (p < 0.001) when compared to standard antibiotic treatment alone. Notably, there was a 50 ± 15% reduction in cell viability and 60 ± 12% reduction in biomass (95% CI) for the 4 biofilms demonstrating the most resistance to antibiotic treatment. 83% of strains tested (n=6) showed decreased bacterial killing by tobramycin at acidic pHs (p < 0.01). However, 25% of strains (n=12) showed improved phage killing at acidic pHs (p < 0.05), with none showing the pattern of reduced efficacy at acidic pH demonstrated by tobramycin. Conclusion: The 4-phage anti-Pa cocktail tested against Pa performs well in pre-formed biofilms and in acidic environments; two conditions intended to mimic the CF lung. To our knowledge, these are the first data looking at the effects of subtle pH changes on phage-mediated bacterial killing in the context of Pa infection. These findings contribute to a growing body of evidence supporting the use of nebulised lytic bacteriophage as a treatment in the context of lung infection.

Keywords: biofilm, cystic fibrosis, pH, Pseudomonas aeruginosa, lytic bacteriophage

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621 Trehalose Application Increased Membrane Stability and Cell Viability to Affect Growth of Wheat Genotypes under Heat Stress

Authors: S. K. Thind, Aparjot Kaur

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Heat stress is one of the major environmental factors drastically reducing wheat production. Crop heat tolerance can be enhanced by preconditioning of plants by exogenous application of osmoprotectants. Presently, the effect of trehalose pretreatment (at 1 mM, and 1.5 nM) under heat stress of 35±2˚C (moderate) and 40±2˚ (severe) for four and eight hour was conducted in wheat (Tricticum aestivum L.) genotypes viz. HD2967, PBW 175, PBW 343, PBW 621, and PBW 590. Heat stress affects wide spectrum of physiological processes within plants that are irreversibly damaged by stress. Membrane thermal stability (MTS) and cell viability was significantly decreased under heat stress for eight hours. Pretreatment with trehalose improved MTS and cell viability under stress and this effect was more promotory with higher concentration. Thermal stability of photosynthetic apparatus differed markedly between genotypes and Hill reaction activity was recorded more in PBW621 followed by C306 as compared with others. In all genotypes photolysis of water showed decline with increase in temperature stress. Trehalose pretreatment helped in sustaining Hill reaction activity probably by stabilizing the photosynthetic apparatus against heat-induced photo inhibition. Both plant growth and development were affected by temperature in both shoot and root under heat stress. The reduction was compensated partially by trehalose (1.5 mM) application. Adaption to heat stress is associated with the metabolic adjustment which led to accumulation of soluble sugars including non-reducing and reducing for their role in adaptive mechanism. Higher acid invertase activity in shoot of tolerant genotypes appeared to be a characteristic for stress tolerance. As sucrose synthase play central role in sink strength and in studied wheat genotype was positively related to dry matter accumulation. The duration of heat stress for eight hours had more severe effect on these parameters and trehalose application at 1.5 mM ameliorated it to certain extent.

Keywords: heat stress, Triticum aestivum, trehalose, membrane thermal stability, triphenyl tetrazolium chloride, reduction test, growth, sugar metabolism

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620 Role of Autophagic Lysosome Reformation for Cell Viability in an in vitro Infection Model

Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

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Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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619 Activity of Resveratrol on the Influence of Aflatoxin B1 on the Testes of Sprague Dawley Rats

Authors: Ali D. Omur, Betul Apaydin Yildirim, Yavuz S. Saglam, Selim Comakli, Mustafa Ozkaraca

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Twenty-eight male Sprague Dawley rats (aged 3 months) were used in the study. The animals were given feed and water as ad libitum. Sprague Dawley rats were randomly divided into 4 groups as 7 rats in each group. Aflatoxin B1 (7.5 μg/200 g), resveratrol (60 mg/kg) was administered to rats in groups other than the control group. At the end of the 16th day, blood, semen and tissue specimens were taken by decapitation under ether anesthesia. The effects of aflatoxin B1 and resveratrol on spermatological, pathological and biochemical parameters were determined in rats. When we evaluate the spermatological parameters, it is understood that resveratrol has a statistically significant difference in terms of sperm motility and viability (membrane integrity) compared to the control group and aflatoxin B1 administration groups, indicating a protective effect on spermatological parameters (groups: control, resveratrol, aflatoxin B1 and Afb1 + res; respectively, values of motility: 71,42 ± 0,52b, 72,85 ± 1, 48c , 60,71 ± 1,30a, 57,14 ± 2, 40a; values of viability: 63,85 ± 1,33b, 70,42 ± 2,61c, 55,00 ± 1,54a, 56,57 ± 0,89a. In terms of pathological parameters -histopathological examination- in the control and resveratrol groups, seminiferous tubules were observed to be in normal structure. In the group treated with aflatoxin, the regular structure of the spermatogenic cells deteriorated, and the seminiferous tubules became necrotic and degenerative. In the group treated with Afb1 + res, the decreasing of necrotic and degenerative changes were determined compared with in the group treated with aflatoxin. As immunohistochemical examination, cleaved caspase 3 expression was found to be very low in the control and resveratrol groups. Cleaved caspase 3 expression was severely exacerbated in seminiferous tubules in aflatoxin group but cleaved caspase 3 expression level decreased in Afb1 + res. In the biochemical direction, resveratrol has been shown to inhibit the adverse effects of aflatoxin on antioxidant levels (GSH-mmol/L, CAT-kU/L, GPx-U/mL, SOD-EU/mL) and to show a protective effect. For this purpose, the use of resveratrol with antioxidant activity was investigated in preventing or ameliorating damage to aflatoxin B1. It has been concluded that resveratrol effectively prevents the aflatoxin-induced testicular damage and lipid peroxidation. It has also been shown that resveratrol has protective effects on sperm motility and viability.

Keywords: Aflatoxin B1, rat, resveratrol, sperm

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618 Cytotoxic Effect of Crude Extract of Sea Pen Virgularia gustaviana on HeLa and MDA-MB-231 Cancer Cell Lines

Authors: Sharareh Sharifi, Pargol Ghavam Mostafavi, Ali Mashinchian Moradi, Mohammad Hadi Givianrad, Hassan Niknejad

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Marine organisms such as soft coral, sponge, ascidians, and tunicate containing rich source of natural compound have been studied in last decades because of their special chemical compounds with anticancer properties. The aim of this study was to investigate anti-cancer property of ethyl acetate extracted from marine sea pen Virgularia gustaviana found from Persian Gulf coastal (Bandar Abbas). The extraction processes were carried out with ethyl acetate for five days. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were used for qualitative identification of crude extract. The viability of HeLa and MDA-Mb-231 cancer cells was investigated using MTT assay at the concentration of 25, 50, and a 100 µl/ml of ethyl acetate is extracted. The crude extract of Virgularia gustaviana demonstrated ten fractions with different Retention factor (Rf) by TLC and Retention time (Rt) evaluated by HPLC. The crude extract dose-dependently decreased cancer cell viability compared to control group. According to the results, the ethyl acetate extracted from Virgularia gustaviana inhibits the growth of cancer cells, an effect which needs to be further investigated in the future studies.

Keywords: anti-cancer, Hela cancer cell, MDA-Md-231 cancer cell, Virgularia gustavina

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617 Viability of Irrigation Water Conservation Practices in the Low Desert of California

Authors: Ali Montazar

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California and the Colorado River Basin are facing increasing uncertainty concerning water supplies. The Colorado River is the main source of irrigation water in the low desert of California. Currently, due to an increasing water-use competition and long-term drought at the Colorado River Basin, efficient use of irrigation water is one of the highest conservation priorities in the region. This study aims to present some of current irrigation technologies and management approaches in the low desert and assess the viability and potential of these water management practices. The results of several field experiments are used to assess five water conservation practices of sub-surface drip irrigation, automated surface irrigation, sprinkler irrigation, tail-water recovery system, and deficit irrigation strategy. The preliminary results of several ongoing studies at commercial fields are presented, particularly researches in alfalfa, sugar beets, kliengrass, sunflower, and spinach fields. The findings indicate that all these practices have significant potential to conserve water (an average of 1 ac-ft/ac) and enhance the efficiency of water use (15-25%). Further work is needed to better understand the feasibility of each of these applications and to help maintain profitable and sustainable agricultural production system in the low desert as water and labor costs, and environmental issues increase.

Keywords: automated surface irrigation, deficit irrigation, low desert of California, sprinkler irrigation, sub-surface drip irrigation, tail-water recovery system

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616 Cryoinjuries in Sperm Cells: Effect of Adaptation of Steps in Cryopreservation Protocol for Boar Semen upon Post-Thaw Sperm Quality

Authors: Aftab Ali

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Cryopreservation of semen is one of the key factors for a successful breeding business along with other factors. To achieve high fertility in boar, one should know about spermatozoa response to different treatments proceeds during cryopreservation. The running project is highly focused on cryopreservation and its effects on sperm quality parameters in both boar and bull semen. Semen sample from A, B, C, and D, were subjected to different thawing conditions and were analyzed upon different treatments in the study. Parameters like sperm cell motility, viability, acrosome, DNA integrity, and phospholipase C zeta were detected by different established methods. Different techniques were used to assess different parameters. Motility was detected using computer assisted sperm analysis, phospholipase C zeta using luminometry while viability, acrosome integrity, and DNA integrity were analyzed using flow cytometry. Thawing conditions were noted to have an effect on sperm quality parameters with motility being the most critical parameter. The results further indicated that the most critical step during cryopreservation of boar semen is when sperm cells are subjected to freezing and thawing. The findings of the present study provide insight that; boar semen cryopreservation is still suboptimal in comparison to bull semen cryopreservation. Thus, there is a need to conduct more research to improve the fertilizing potential of cryopreserved boar semen.

Keywords: cryopreservation, computer assisted sperm, flow cytometry, luminometry

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615 Evaluating Viability of Solar Tubewell Irrigation Technology

Authors: Junaid N. Chauhdary, Bernard A. Engel, Allah Bakhsh

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Solar powered tubewells can be a reliable and affordable source of supplying irrigation water compared with electric or diesel operated tubewells due to frequent load shedding and soaring energy prices. A study was conducted on a solar tubewell installed at the Water Management Research Center (WMRC), University of Agriculture, Faisalabad to investigate the viability of a solar powered tubewell in terms of discharge and benefit cost ratio. The tubewell discharge was 50 m3hr-1 with a total dynamic head of 30 m. The depth of bore was 31 m (14 m blind + 17 m screen) with a casing diameter of 15.2 cm (6 inches). A 3-stage submersible pump of 10.2 cm (4 inch) diameter was lowered in the casing to a depth of 22 m. The pump was powered from 21 solar panels of 200 W capacity each. The tubewell peak discharge was observed as 6 and 7 hr day-1 in winter and summer, respectively. The breakeven analysis of the solar tubewell showed that the payback period of the solar tubewell was 1.5 years of its 10 year usable life with an IRR (internal rate of return) of 69 %. The BCR (benefit cost ratio) of the solar tubewell at 2, 4, 6, and 8 percent discount rate were 3.75, 3.45, 3.19 and 2.96, respectively. The NPV (net present value) of the solar tubewell at 2, 4, 6, and 8 % discount rates were 1.89, 1.65, 1.45 and 1.27 million rupees, respectively. These results indicated that the solar powered tubewells are a viable option as well as environmentally friendly and can be adopted by the farmers due to their affordable payback period.

Keywords: benefit cost ratio, internal rate of return (IRR), net present value (NPV), solar tubewell

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614 Indirect Solar Desalination: Value Engineering and Cost Benefit Analysis

Authors: Grace Rachid, Mutasem El Fadel, Mahmoud Al Hindi, Ibrahim Jamali, Daniel Abdel Nour

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This study examines the feasibility of indirect solar desalination in oil producing countries in the Middle East and North Africa (MENA) region. It relies on value engineering (VE) and cost-benefit with sensitivity analyses to identify optimal coupling configurations of desalination and solar energy technologies. A comparative return on investment was assessed as a function of water costs for varied plant capacities (25,000 to 75,000 m3/day), project lifetimes (15 to 25 years), and discount rates (5 to 15%) taking into consideration water and energy subsidies, land cost as well as environmental externalities in the form of carbon credit related to greenhouse gas (GHG) emissions reduction. The results showed reverse osmosis (RO) coupled with photovoltaic technologies (PVs) as the most promising configuration, robust across different prices for Brent oil, discount rates, as well as different project lifetimes. Environmental externalities and subsidies analysis revealed that a 16% reduction in existing subsidy on water tariffs would ensure economic viability. Additionally, while land costs affect investment attractiveness, the viability of RO coupled with PV remains possible for a land purchase cost < $ 80/m2 or a lease rate < $1/m2/yr. Beyond those rates, further subsidy lifting is required.

Keywords: solar energy, desalination, value engineering, CBA, carbon credit, subsidies

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613 Assessment of Selected Marine Organisms from Malaysian Coastal Areas for Inhibitory Activity against the Chikungunya Virus

Authors: Yik Sin Chan, Nam Weng Sit, Fook Yee Chye, van Ofwegen Leen, de Voogd Nicole, Kong Soo Khoo

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Chikungunya fever is an arboviral disease transmitted by the Aedes mosquitoes. It has resulted in epidemics of the disease in tropical countries in the Indian Ocean and South East Asian regions. The recent spread of this disease to the temperate countries such as France and Italy, coupled with the absence of vaccines and effective antiviral drugs make chikungunya fever a worldwide health threat. This study aims to investigate the anti-chikungunya virus activity of selected marine organism samples collected from Malaysian coastal areas, including seaweeds (Caulerpa racemosa, Caulerpa sertularioides and Kappaphycus alvarezii), a soft coral (Lobophytum microlobulatum) and a sponge (Spheciospongia vagabunda). Following lyophilization (oven drying at 40C for K. alvarezii) and grinding to powder form, each sample was subjected to sequential solvent extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and distilled water in order to extract bioactive compounds. The antiviral activity was evaluated using monkey kidney epithelial (Vero) cells infected with the virus (multiplicity of infection=1). The cell viability was determined by Neutral Red uptake assay. 70% of the 30 extracts showed weak inhibitory activity with cell viability ≤30%. Seven of the extracts exhibited moderate inhibitory activity (cell viability: 31%-69%). These were the chloroform, ethyl acetate, ethanol and methanol extracts of C. racemosa; chloroform and ethyl acetate extracts of L. microlobulatum; and the chloroform extract of C. sertularioides. Only the hexane and ethanol extracts of L. microlobulatum showed strong inhibitory activity against the virus, resulting in cell viabilities (mean±SD; n=3) of 73.3±2.6% and 79.2±0.9%, respectively. The corresponding mean 50% effective concentrations (EC50) for the extracts were 14.2±0.2 and 115.3±1.2 µg/mL, respectively. The ethanol extract of the soft coral L. microlobulatum appears to hold the most promise for further characterization of active principles as it possessed greater selectivity index (SI>5.6) compared to the hexane extract (SI=2.1).

Keywords: antiviral, seaweed, sponge, soft coral, vero cell

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612 Viability of Rice Husk Ash Concrete Brick/Block from Green Electricity in Bangladesh

Authors: Mohammad A. N. M. Shafiqul Karim

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As a developing country, Bangladesh has to face numerous challenges. Self Independence in electricity, contributing to climate change by reducing carbon emission and bringing the backward population of society to the mainstream is more challenging for them. Therefore, it is essential to ensure recycled use of local products to the maximum level in every sector. Some private organizations have already worked alongside government to bring the backward population to the mainstream by developing their financial capacities. As rice husk is the largest single category of the total energy supply in Bangladesh. As part of this strategy, rice husk can play a great as a promising renewable energy source, which is readily available, has considerable environmental benefits and can produce electricity and ensure multiple uses of byproducts in construction technology. For the first time in Bangladesh, an experimental multidimensional project depending on Rice Husk Electricity and Rice Husk Ash (RHA) concrete brick/block under Green Eco-Tech Limited has already been started. Project analysis, opportunity, sustainability, the high monitoring component, limitations and finally evaluated data reflecting the viability of establishing more projects using rice husk are discussed in this paper. The by-product of rice husk from the production of green electricity, RHA, can be used for making, in particular, RHA concrete brick/block in Bangladeshi aspects is also discussed here.

Keywords: project analysis, rice husk, rice husk ash concrete brick/block, compressive strength of rice husk ash concrete brick/block

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611 A Comparative Analysis of Solid Waste Treatment Technologies on Cost and Environmental Basis

Authors: Nesli Aydin

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Waste management decision making in developing countries has moved towards being more pragmatic, transparent, sustainable and comprehensive. Turkey is required to make its waste related legislation compatible with European Legislation as it is a candidate country of the European Union. Improper Turkish practices such as open burning and open dumping practices must be abandoned urgently, and robust waste management systems have to be structured. The determination of an optimum waste management system in any region requires a comprehensive analysis in which many criteria are taken into account by stakeholders. In conducting this sort of analysis, there are two main criteria which are evaluated by waste management analysts; economic viability and environmentally friendliness. From an analytical point of view, a central characteristic of sustainable development is an economic-ecological integration. It is predicted that building a robust waste management system will need significant effort and cooperation between the stakeholders in developing countries such as Turkey. In this regard, this study aims to provide data regarding the cost and environmental burdens of waste treatment technologies such as an incinerator, an autoclave (with different capacities), a hydroclave and a microwave coupled with updated information on calculation methods, and a framework for comparing any proposed scenario performances on a cost and environmental basis.

Keywords: decision making, economic viability, environmentally friendliness, waste management systems

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610 Green Hydrogen: Exploring Economic Viability and Alluring Business Scenarios

Authors: S. Sakthivel

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Currently, the global economy is based on the hydrocarbon economy, which is referencing the global hydrocarbon industry. Problems of using these fossil fuels (like oil, NG, coal) are emitting greenhouse gases (GHGs) and price fluctuation, supply/distribution, etc. These challenges can be overcome by using clean energy as hydrogen. The hydrogen economy is the use of hydrogen as a low carbon fuel, particularly for hydrogen vehicles, alternative industrial feedstock, power generation, and energy storage, etc. Engineering consulting firms have a significant role in this ambition and green hydrogen value chain (i.e., integration of renewables, production, storage, and distribution to end-users). Typically, the cost of green hydrogen is a function of the price of electricity needed, the cost of the electrolyser, and the operating cost to run the system. This article focuses on economic viability and explores the alluring business scenarios globally. Break-even analysis was carried out for green hydrogen production and in order to evaluate and compare the impact of the electricity price on the production costs of green hydrogen and relate it to fossil fuel-based brown/grey/blue hydrogen costs. It indicates that the cost of green hydrogen production will fall drastically due to the declining costs of renewable electricity prices and along with the improvement and scaling up of electrolyser manufacturing. For instance, in a scenario where electricity prices are below US$ 40/MWh, green hydrogen cost is expected to reach cost competitiveness.

Keywords: green hydrogen, cost analysis, break-even analysis, renewables, electrolyzer

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609 A Project Screening System for Energy Enterprise Based on Dempster-Shafer Theory

Authors: Woosik Jang, Seung Heon Han, Seung Won Baek

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Natural gas (NG) is an energy resource in a few countries, and most NG producers do business in politically unstable countries. In addition, as 90% of the LNG market is controlled by a small number of international oil companies (IOCs) and national oil companies (NOCs), entry of latecomers into the market is extremely limited. To meet these challenges, project viability needs to be assessed based on limited information from a project screening perspective. However, the early stages of the project have the following difficulties: (1) What are the factors to consider? (2) How many professionals do you need to decide? (3) How to make the best decision with limited information? To address this problem, this study proposes a model for evaluating LNG project viability based on the Dempster-Shafer theory (DST). A total of 11 indicators for analyzing the gas field, reflecting the characteristics of the LNG industry, and 23 indicators for analyzing the market environment, were identified. The proposed model also evaluates the LNG project based on the survey and provides uncertainty of the results based on DST as well as quantified results. Thus, the proposed model is expected to be able to support the decision-making process of the gas field project using quantitative results as a systematic framework, and it was developed as a stand-alone system to improve its usefulness in practice. Consequently, the amount of information and the mathematical approach are expected to improve the quality and opportunity of decision making for LNG projects for enterprises.

Keywords: project screen, energy enterprise, decision support system, Dempster-Shafer theory

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608 Mothwash Formulation of Moringa Leaf (Moringa Oleifera) and Its Activity as an Antibacterial for Streptococcus Mutans

Authors: Amalia Dwi Berliyanti Amel

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Streptococcus mutants bacteria are bacteria that are believed to be the cause of the growth of dental plaque which can further adversely affect dental caries if left unchecked. Previous research has shown that Moringa leaf extract can slow down the growth rate of this bacterium. This study aims to make the best formulation of mouthwash with the active ingredient of Moringa leaf extract based on its antibacterial and organoleptic test results. Nine mouthwash variations were carried out with two factors and three levels, namely a comparison of the concentration of sorbitol (A) with three levels namely 15% (A1), 20% (A2), and 25% (A3), and peppermint added (B) with three levels, namely 0.2% (B1), 0.25% (B2), and 0.3% (B3). The test parameters performed as the determination of the best mouthwash are based on physicochemical properties which include pH and viscosity as well as organoleptic test results which include color, viscosity, aroma, taste, sensation in the mouth, and general appearance. The results showed that the bright zone as a test for the antibacterial activity of Streptococcus mutants began to be seen at a concentration of 5%. Moringa leaf mouthwash formulation has a pH value between 6 - 7, with a control of 6. Whereas the mucosa leaf mouthwash vascularity produced between 1.1 - 1.7 cP with a control of 1.1 cP. Moringa leaf mouthwash and control have the same total number of microbes, namely 0 colonies / mL. Based on organoleptic tests performed with 20 panelists, it was shown that the best mouthwash formulation was formulation A1B3 with sorbitol composition 15% and peppermint 0.3%.

Keywords: antibasteria, formula, moringa leaf, mouthwash

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607 Anti-Melanogenesis and Anti-Inflammatory Effects of Opuntia humifusa

Authors: Yonghwa Lee, Yoon Suk Kim, Yongsub Yi

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This study was to confirm the effects of anti-melanogenesis and anti-inflammatory effects from Opuntia humifusa fruit and stem extracts. A potent anti-oxidant activity was shown from the leaf extract at IC50 value of 38.33±1.07 μg/mL and fruit extract at IC50 value of 40.23±2.21 μg/mL by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Also, phenolic contents were confirmed total phenolic assay by high performance liquid chromatography (HPLC). Fraction of taxifolin from leaf extract was identified using HPLC and gas chromatography/mass spectrometry. The extracts of Opuntia humifusa fruit and stem were confirmed about toxicity effect in B16 F1 by cell viability. Melanin contents were decreased. Opuntia humifusa fruit and stem extracts had a positive effect of melanin synthesis inhibition for skin whitening. In investigating the anti-inflammatory activities of Opuntia humifusa, the results of cell viability indicated that taxifolin did not show cytotoxicity on RAW264.7 cells at 500 μM of concentration. The results show that taxifolin inhibited lipopolysaccharide (LPS)-induced production of Nitrite oxide (NO). In addition, taxifolin indicated the inhibition of lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF) -α and interleukin (IL) -6 productions by cytokine assay and cyclooxygenase (COX)-2 expression by western blot analysis, meaning that taxifolin had a significant anti-inflammatory effect. Our results suggested that taxifolin from Opuntia humifusa has anti-melanogenesis and anti-inflammatory activities.

Keywords: anti-melanogenesis, anti-inflammatory, Opuntia humifusa, taxifolin

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606 Evaluation of the Influence of Graphene Oxide on Spheroid and Monolayer Culture under Flow Conditions

Authors: A. Zuchowska, A. Buta, M. Mazurkiewicz-Pawlicka, A. Malolepszy, L. Stobinski, Z. Brzozka

Abstract:

In recent years, graphene-based materials are finding more and more applications in biological science. As a thin, tough, transparent and chemically resistant materials, they appear to be a very good material for the production of implants and biosensors. Interest in graphene derivatives also resulted at the beginning of research about the possibility of their application in cancer therapy. Currently, the analysis of their potential use in photothermal therapy and as a drug carrier is mostly performed. Moreover, the direct anticancer properties of graphene-based materials are also tested. Nowadays, cytotoxic studies are conducted on in vitro cell culture in standard culture vessels (macroscale). However, in this type of cell culture, the cells grow on the synthetic surface in static conditions. For this reason, cell culture in macroscale does not reflect in vivo environment. The microfluidic systems, called Lab-on-a-chip, are proposed as a solution for improvement of cytotoxicity analysis of new compounds. Here, we present the evaluation of cytotoxic properties of graphene oxide (GO) on breast, liver and colon cancer cell line in a microfluidic system in two spatial models (2D and 3D). Before cell introduction, the microchambers surface was modified by the fibronectin (2D, monolayer) and poly(vinyl alcohol) (3D, spheroids) covering. After spheroid creation (3D) and cell attachment (2D, monolayer) the selected concentration of GO was introduced into microsystems. Then monolayer and spheroids viability/proliferation using alamarBlue® assay and standard microplate reader was checked for three days. Moreover, in every day of the culture, the morphological changes of cells were determined using microscopic analysis. Additionally, on the last day of the culture differential staining using Calcein AM and Propidium iodide were performed. We were able to note that the GO has an influence on all tested cell line viability in both monolayer and spheroid arrangement. We showed that GO caused higher viability/proliferation decrease for spheroids than a monolayer (this was observed for all tested cell lines). Higher cytotoxicity of GO on spheroid culture can be caused by different geometry of the microchambers for 2D and 3D cell cultures. Probably, GO was removed from the flat microchambers for 2D culture. Those results were also confirmed by differential staining. Comparing our results with the studies conducted in the macroscale, we also proved that the cytotoxic properties of GO are changed depending on the cell culture conditions (static/ flow).

Keywords: cytotoxicity, graphene oxide, monolayer, spheroid

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605 Towards the Modeling of Lost Core Viability in High-Pressure Die Casting: A Fluid-Structure Interaction Model with 2-Phase Flow Fluid Model

Authors: Sebastian Kohlstädt, Michael Vynnycky, Stephan Goeke, Jan Jäckel, Andreas Gebauer-Teichmann

Abstract:

This paper summarizes the progress in the latest computational fluid dynamics research towards the modeling in of lost core viability in high-pressure die casting. High-pressure die casting is a process that is widely employed in the automotive and neighboring industries due to its advantages in casting quality and cost efficiency. The degrees of freedom are however somewhat limited as it has been so far difficult to use lost cores in the process. This is right now changing and the deployment of lost cores is considered a future growth potential for high-pressure die casting companies. The use of this technology itself is difficult though. The strength of the core material, as chiefly salt is used, is limited and experiments have shown that the cores will not hold under all circumstances and process designs. For this purpose, the publicly available CFD library foam-extend (OpenFOAM) is used, and two additional fluid models for incompressible and compressible two-phase flow are implemented as fluid solver models into the FSI library. For this purpose, the volume-of-fluid (VOF) methodology is used. The necessity for the fluid-structure interaction (FSI) approach is shown by a simple CFD model geometry. The model is benchmarked against analytical models and experimental data. Sufficient agreement is found with the analytical models and good agreement with the experimental data. An outlook on future developments concludes the paper.

Keywords: CFD, fluid-structure interaction, high-pressure die casting, multiphase flow

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604 Effects of Gelatin on Characteristics and Dental Pathogen Inhibition by Silver Nanoparticles Synthesized from Ascorbic Acid

Authors: Siriporn Okonogi, Temsiri Suwan, Sakornrat Khongkhunthian, Jakkapan Sirithunyalug

Abstract:

In this study, silver nanoparticles (AgNPs) were prepared using ascorbic acid as a reducing agent and silver nitrate as a precursor. The effects of gelatin (G) on particle characteristics and dental pathogen inhibition were investigated. The spectra of AgNPs and G-AgNPs were compared using UV-Vis and Energy-dispersive X-ray (EDX) spectroscopy. The obtained AgNPs and G-AgNPs showed the maximum absorption at 410 and 430 nm, respectively, and EDX spectra of both systems confirmed Ag element. Scanning electron microscope showed that AgNPs and G-AgNPs were spherical in shape. Particles size, size distribution, and zeta potential were determined using dynamic light scattering approach. The size of AgNPs and G-AgNPs were 56 ± 2.4 and 67 ± 3.6 nm, respectively with a size distribution of 0.23 ± 0.03 and 0.19 ± 0.02, respectively. AgNPs and G-AgNPs exhibited negative zeta potential of 24.1 ± 2.7 mV and 32.7 ± 1.2 mV, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the obtained AgNPs and G-AgNPs against three strains of dental pathogenic bacteria; Streptococcus gordonii, Streptococcus mutans, and Staphylococcus aureus were determined using broth dilution method. AgNPs and G-AgNPs showed the strongest inhibition against S. gordonii with the MIC of 0.05 and 0.025 mg/mL, respectively and the MBC of 0.1 and 0.05 mg/mL, respectively. Cytotoxicity test of AgNPs and G-AgNPs on human breast cancer cells using MTT assay indicated that G-AgNPs (0.1 mg/mL) was significantly stronger toxic than AgNPs with the cell inhibition of 91.1 ± 5.4%. G-AgNPs showed significantly less aggregation after storage at room temperature for 90 days than G-AgNPs.

Keywords: antipathogenic activity, ascorbic acid, cytotoxicity, stability

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603 Effect of Feed Additive on Cryopreservation of Barki Ram Semen

Authors: Abdurzag Kerban, Mostfa M. Abou-Ahmed, Abdelrof M. Ghallab, Mona H. Shaker

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Preservation of semen had a major impact on sheep genetic breeding. The aim of this study was to evaluate the effect of protected fat, probiotic and zinc-enriched diets on semen freezability. Twenty two Barki rams were randomly assigned into four groups; Group I (n=5) was fed the basal diet enriched with 3.7% of dry fat/kg concentration/day, Group II (n=5) was fed a basal diet-enriched with 10gm of probiotic /head/day, Group III (n=6) was fed on the basal diet enriched with 100 ppm of 10% zinc chelated with methionine/kg dry matter/day and Group IV (n=6) was served as control. A pool of three to four ejaculates were pooled from rams within a period of ten weeks. Semen was diluted in egg yolk-Tris diluent and processed in 0.25 ml straw. Motility was evaluated after dilution, before freezing and post-thawing at 0, 1, 2 and 3 hour incubation. Viability index, acrosome integrity and leakage of intracellular enzymes (Aspartat aminotransferase and Alkline phosphatase) were also evaluated. Spermatozoa exhibited highly significant (P<0.01) percentages of motility at 0, 1, 2, and 3 hours incubation after thawing, viability index and acrosome integrity in rams fed a diet enriched with protected fat and zinc groups as compared with probiotic and control groups. Also, the mean value of extracellular leakage of AST was significantly lower in fat and zinc group as compared with probiotic and control groups. In conclusion, semen freezability was improved in animals fed a diet fortified with fat and zinc with no significant improvement in animals fed the probiotic-enriched diet.

Keywords: Barki ram semen, freezing, straw, feed additives

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602 Biofuels from Hybrid Poplar: Using Biochemicals and Wastewater Treatment as Opportunities for Early Adoption

Authors: Kevin W. Zobrist, Patricia A. Townsend, Nora M. Haider

Abstract:

Advanced Hardwood Biofuels Northwest (AHB) is a consortium funded by the United States Department of Agriculture (USDA) to research the potential for a system to produce advanced biofuels (jet fuel, diesel, and gasoline) from hybrid poplar in the Pacific Northwest region of the U.S. An Extension team was established as part of the project to examine community readiness and willingness to adopt hybrid as a purpose-grown bioenergy crop. The Extension team surveyed key stakeholder groups, including growers, Extension professionals, policy makers, and environmental groups, to examine attitudes and concerns about growing hybrid poplar for biofuels. The surveys found broad skepticism about the viability of such a system. The top concern for most stakeholder groups was economic viability and the availability of predictable markets. Growers had additional concerns stemming from negative past experience with hybrid poplar as an unprofitable endeavor for pulp and paper production. Additional barriers identified included overall land availability and the availability of water and water rights for irrigation in dry areas of the region. Since the beginning of the project, oil and natural gas prices have plummeted due to rapid increases in domestic production. This has exacerbated the problem with economic viability by making biofuels even less competitive than fossil fuels. However, the AHB project has identified intermediate market opportunities to use poplar as a renewable source for other biochemicals produced by petroleum refineries, such as acetic acid, ethyl acetate, ethanol, and ethylene. These chemicals can be produced at a lower cost with higher yields and higher, more-stable prices. Despite these promising market opportunities, the survey results suggest that it will still be challenging to induce growers to adopt hybrid poplar. Early adopters will be needed to establish an initial feedstock supply for a budding industry. Through demonstration sites and outreach events to various stakeholder groups, the project attracted interest from wastewater treatment facilities, since these facilities are already growing hybrid poplar plantations for applying biosolids and treated wastewater for further purification, clarification, and nutrient control through hybrid poplar’s phytoremediation capabilities. Since these facilities are already using hybrid poplar, selling the wood as feedstock for a biorefinery would be an added bonus rather than something requiring a high rate of return to compete with other crops and land uses. By holding regional workshops and conferences with wastewater professionals, AHB Extension has found strong interest from wastewater treatment operators. In conclusion, there are several significant barriers to developing a successful system for producing biofuels from hybrid poplar, with the largest barrier being economic viability. However, there is potential for wastewater treatment facilities to serve as early adopters for hybrid poplar production for intermediate biochemicals and eventually biofuels.

Keywords: hybrid poplar, biofuels, biochemicals, wastewater treatment

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601 Antibacterial Activity and Cytotoxicity of Silver Nanoparticles Synthesized by Moringa oleifera Extract as Reducing Agent

Authors: Temsiri Suwan, Penpicha Wanachantararak, Sakornrat Khongkhunthian, Siriporn Okonogi

Abstract:

In the present study, silver nanoparticles (AgNPs) were synthesized by green synthesis approach using Moringa oleifera aqueous extract (ME) as a reducing agent and silver nitrate as a precursor. The obtained AgNPs were characterized using UV-Vis spectroscopy (UV-Vis), dynamic light scattering (DLS), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray diffractometry (XRD). The results from UV-Vis revealed that the maximum absorption of AgNPs was at 430 nm and the EDX spectrum confirmed Ag element. The results from DLS indicated that the amount of ME played an important role in particle size, size distribution, and zeta potential of the obtained AgNPs. The smallest size (62.4 ± 1.8 nm) with narrow distribution (0.18 ± 0.02) of AgNPs was obtained after using 1% w/v of ME. This system gave high negative zeta potential of -36.5 ± 2.8 mV. SEM results indicated that the obtained AgNPs were spherical in shape. Antibacterial activity using dilution method revealed that the minimum inhibitory and minimum bactericidal concentrations of the obtained AgNPs against Streptococcus mutans were 0.025 and 0.1 mg/mL, respectively. Cytotoxicity test of AgNPs on adenocarcinomic human alveolar basal epithelial cells (A549) indicated that the particles impacted against A549 cells. The percentage of cell growth inhibition was 87.5 ± 3.6 % when only 0.1 mg/mL AgNPs was used. These results suggest that ME is the potential reducing agent for green synthesis of AgNPs.

Keywords: antibacterial activity, Moringa oleifera extract, reducing agent, silver nanoparticles

Procedia PDF Downloads 94
600 Farmers Willingness to Pay for Irrigated Maize Production in Rural Kenya

Authors: Dennis Otieno, Lilian Kirimi Nicholas Odhiambo, Hillary Bii

Abstract:

Kenya is considered to be a middle level income country and usuaaly does not meet household food security needs especially in North and South eastern parts. Approximately half of the population is living under the poverty line (www, CIA 1, 2012). Agriculture is the largest sector in the country, employing 80% of the population. These are thereby directly dependent on the sufficiency of outputs received. This makes efficient, easy-accessible and cheap agricultural practices an important matter in order to improve food security. Maize is the prime staple food commodity in Kenya and represents a substantial share of people’s nutritional intake. This study is the result of questionnaire based interviews, Key informant and focus group discussion involving 220 small scale maize farmers Kenyan. The study was located to two separated areas; Lower Kuja, Bunyala, Nandi, Lower Nzoia, Perkerra, Mwea Bura, Hola and Galana Kulalu in Kenya. The questionnaire captured the farmers’ use and perceived importance of the use irrigation services and irrigated maize production. Viability was evaluated using the four indices which were all positive with NPV giving positive cash flows in less than 21 years at most for one season output. The mean willingness to pay was found to be KES 3082 and willingness to pay increased with increase in irrigation premiums. The economic value of water was found to be greater than the willingness to pay implying that irrigated maize production is sustainable. Farmers stated that viability was influenced by high output levels, good produce quality, crop of choice, availability of sufficient water and enforcement the last two factors had a positive influence while the other had negative effect on the viability of irrigated maize. A regression was made over the correlation between the willingness to pay for irrigated maize production using scheme and plot level factors. Farmers that already use other inputs such as animal manure, hired labor and chemical fertilizer should also have a demand for improved seeds according to Liebig's law of minimum and expansion path theory. The regression showed that premiums, and high yields have a positive effect on willingness to pay while produce quality, efficient fertilizer use, and crop season have a negative effect.

Keywords: maize, food security, profits, sustainability, willingness to pay

Procedia PDF Downloads 198
599 An Assessment of Financial Viability and Sustainability of Hydroponics Using Reclaimed Water Using LCA and LCC

Authors: Muhammad Abdullah, Muhammad Atiq Ur Rehman Tariq, Faraz Ul Haq

Abstract:

In developed countries, sustainability measures are widely accepted and acknowledged as crucial for addressing environmental concerns. Hydroponics, a soilless cultivation technique, has emerged as a potentially sustainable solution as it can reduce water consumption, land use, and environmental impacts. However, hydroponics may not be economically viable, especially when using reclaimed water, which may entail additional costs and risks. This study aims to address the critical question of whether hydroponics using reclaimed water can achieve a balance between sustainability and financial viability. Life Cycle Assessment (LCA) and Life Cycle Cost (LCC) will be integrated to assess the potential of hydroponics whether it is environmentally sustainable and economically viable. Life cycle assessment, or LCA, is a methodology for assessing environmental impacts associated with all the stages of the life cycle of a commercial product, process, or service. While Life Cycle Cost (LCC) is an approach that assesses the total cost of an asset over its life cycle, including initial capital costs and maintenance costs. The expected benefits of this study include supporting evidence-based decision-making for policymakers, farmers, and stakeholders involved in agriculture. By quantifying environmental impacts and economic costs, this research will facilitate informed choices regarding the adoption of hydroponics with reclaimed water. It is believed that the outcomes of this research work will help to achieve a sustainable approach to agricultural production, aligning with sustainability goals while considering economic factors by adopting hydroponic technique.

Keywords: hydroponic, life cycle assessment, life cycle cost, sustainability

Procedia PDF Downloads 54
598 Evaluation of Chemical Compositions and Biological Activities of Five Essential Oils

Authors: G. Ozturk, B. Demirci

Abstract:

It is well known that essential oils used for therapeutic purposes for many years. In this study, five different Pharmacopoeia grade essential oils (Achillea millefolium L., Pimpinella anisum L., Matricaria recutita L., Eucalyptus globulus L., Salvia officinalis L.) which obtained from commercial sources were evaluated for chemical compositions, synergistic antimicrobial activities, and lipoxygenase enzyme inhibitions. Volatile components were determined by gas chromatography/flame ionization detector and gas chromatography/mass spectrometer, simultaneously. The potential antimicrobial activity of essential oils was tested against oral pathogenic standard strains such as Streptococcus mutans, Streptococcus sanguinis, Staphylococcus aureus, Corynebacterium striatum, Candida albicans and Candida krusei by broth microdilution methods. Ciprofloxacin and ketoconazole were used positive controls. It has been observed that the essential oils tested have average inhibitory antimicrobial activity against oral pathogens with a Minimum Inhibition Concentration of 20-0.625 mg/mL. The active essential oils have been combined with antibiotics and synergistic effects have been evaluated by Checkerboard method. ƩFIC values were determined. In combination with antibiotics M. recutita essential oil has been shown to have a synergistic effect against S. aureus in combination with tetracycline (ƩFIC 0.46). In addition, 5-LOX inhibitory activity was measured by modifying the spectrophotometric method developed by Baylac and Racine. As a result, 5-LOX % inhibition of S. officinalis, E. globulus and M. recutita were calculated as 34.0 ± 6.66, 72.7 ± 2.78 and 27.7 ± 0.60, respectively.

Keywords: antimicrobial activity, essential oils, synergistic activity, 5-lipoxygenase inhibition

Procedia PDF Downloads 84
597 Configuring Systems to Be Viable in a Crisis: The Role of Intuitive Decision-Making

Authors: Ayham Fattoum, Simos Chari, Duncan Shaw

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Volatile, uncertain, complex, and ambiguous (VUCA) conditions threaten systems viability with emerging and novel events requiring immediate and localized responses. Such responsiveness is only possible through devolved freedom and emancipated decision-making. The Viable System Model (VSM) recognizes the need and suggests maximizing autonomy to localize decision-making and minimize residual complexity. However, exercising delegated autonomy in VUCA requires confidence and knowledge to use intuition and guidance to maintain systemic coherence. This paper explores the role of intuition as an enabler of emancipated decision-making and autonomy under VUCA. Intuition allows decision-makers to use their knowledge and experience to respond rapidly to novel events. This paper offers three contributions to VSM. First, it designs a system model that illustrates the role of intuitive decision-making in managing complexity and maintaining viability. Second, it takes a black-box approach to theory development in VSM to model the role of autonomy and intuition. Third, the study uses a multi-stage discovery-oriented approach (DOA) to develop theory, with each stage combining literature, data analysis, and model/theory development and identifying further questions for the subsequent stage. We synthesize literature (e.g., VSM, complexity management) with seven months of field-based insights (interviews, workshops, and observation of a live disaster exercise) to develop a framework of intuitive complexity management framework and VSM models. The results have practical implications for enhancing the resilience of organizations and communities.

Keywords: Intuition, complexity management, decision-making, viable system model

Procedia PDF Downloads 51
596 Wind Power Mapping and NPV of Embedded Generation Systems in Nigeria

Authors: Oluseyi O. Ajayi, Ohiose D. Ohijeagbon, Mercy Ogbonnaya, Ameh Attabo

Abstract:

The study assessed the potential and economic viability of stand-alone wind systems for embedded generation, taking into account its benefits to small off-grid rural communities at 40 meteorological sites in Nigeria. A specific electric load profile was developed to accommodate communities consisting of 200 homes, a school and a community health centre. This load profile was incorporated within the distributed generation analysis producing energy in the MW range, while optimally meeting daily load demand for the rural communities. Twenty-four years (1987 to 2010) of wind speed data at a height of 10m utilized for the study were sourced from the Nigeria Meteorological Department, Oshodi. The HOMER® software optimizing tool was engaged for the feasibility study and design. Each site was suited to 3MW wind turbines in sets of five, thus 15MW was designed for each site. This design configuration was adopted in order to easily compare the distributed generation system amongst the sites to determine their relative economic viability in terms of life cycle cost, as well as levelised cost of producing energy. A net present value was estimated in terms of life cycle cost for 25 of the 40 meteorological sites. On the other hand, the remaining sites yielded a net present cost; meaning the installations at these locations were not economically viable when utilizing the present tariff regime for embedded generation in Nigeria.

Keywords: wind speed, wind power, distributed generation, cost per kilowatt-hour, clean energy, Nigeria

Procedia PDF Downloads 379
595 Sensitivity of Acanthamoeba castellanii-Grown Francisella to Three Different Disinfectants

Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

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Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

Procedia PDF Downloads 77
594 Assessing Storage of Stability and Mercury Reduction of Freeze-Dried Pseudomonas putida within Different Types of Lyoprotectant

Authors: A. A. M. Azoddein, Y. Nuratri, A. B. Bustary, F. A. M. Azli, S. C. Sayuti

Abstract:

Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage in 4oC without vacuum. Polyethylene glycol (PEG) pre-treated freeze dry cells and broth pre-treated freeze dry cells after freeze-dry recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introduce freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 56.78% and 17.91%. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks were 26.35% and 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been growth in agar. Result from this study may be beneficial and useful as initial reference before commercialize freeze-dried P. putida.

Keywords: Pseudomonas putida, freeze-dry, PEG, tween80/Sucrose, mercury, cell viability

Procedia PDF Downloads 339