Search results for: broth micro dilution assay

Authors: Ashenafi Sileshi Etefa, Bezawit Worku Yenealem

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Micro-insurance is a relatively new concept that is just being introduced in Ethiopia. For an agrarian economy dominated by small holder farming and vulnerable to natural disasters, mainly drought, the need for an Index-Based Micro Insurance (IBMI) is crucial. Since IBMI solves moral hazard, adverse selection, and access issues to poor clients, it is preferable over traditional insurance products. IBMI is being piloted in drought prone areas of Ethiopia with the aim of learning and expanding the service across the country. This article analyses the demand of IBMI and the barriers to demand and finds that the demand for IBMI has so far been constrained by lack of awareness, trust issues, costliness, and the level of basis risk; and recommends reducing the basis risk and increasing the role of government and farmer cooperatives.

Keywords: agriculture, index based micro-insurance (IBMI), drought, micro-finance institution (MFI)

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Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer

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The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.

Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor

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Authors: Megersa Dugasa Fite

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This paper analyses regulatory and supervisory issues in the Ethiopian micro finance sector, which caters to the needs of those who have been excluded from the formal financial sector. Micro-finance has received increased importance in development because of its grand goal to give credits to the poor to raise their economic and social well-being and improve the quality of lives. The micro-finance at present has been moving towards a credit-plus period through covering savings and insurance functions. It thus helps in reducing the rate of financial exclusion and social segregation, alleviating poverty and, consequently, stimulating development. The Ethiopian micro finance policy has been generally positive and developmental but major regulatory and supervisory limitations such as the absolute prohibition of NGOs to participate in micro credit functions, higher risks for depositors of micro-finance institutions, lack of credit information services with research and development, the unmet demand, and risks of market failures due to over-regulation are disappointing. Therefore, to remove the limited reach and high degree of problems typical in the informal means of financial intermediation plus to deal with the failure of formal banks to provide basic financial services to a significant portion of the country’s population, more needs to be done on micro finance. Certain key regulatory and supervisory revisions hence need to be taken to strengthen the Ethiopian micro finance sector so that it can practically provide majority poor access to a range of high quality financial services that help them work their way out of poverty and the incapacity it imposes.

Keywords: micro-finance, micro-finance regulation and supervision, micro-finance institutions, financial access, social segregation, poverty alleviation, development, Ethiopia

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Authors: Emi Govorčin Bajsića, Vesna Ocelić Bulatović, Miroslav Slouf, Ana Šitum

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Composites based on a biodegradable polycaprolactone (PCL) containing 0.5, 1.0 and 2.0 wt % of titanium dioxide (TiO2) micro and nanoparticles were prepared by melt mixing and the effect of filler type and contents on the thermal properties, dynamic-mechanical behaviour and morphology were investigated. Measurements of storage modulus and loss modulus by dynamic mechanical analysis (DMA) showed better results for microfilled PCL/TiO2 composites than nanofilled composites, with the same filler content. DSC analysis showed that the Tg and Tc of micro and nanocomposites were slightly lower than those of neat PCL. The crystallinity of the PCL increased with the addition of TiO2 micro and nanoparticles; however, the c for the PCL was unchanged with micro TiO2 content. The thermal stability of PCL/TiO2 composites were characterized using thermogravimetric analysis (TGA). The initial weight loss (5 wt %) occurs at slightly higher temperature with micro and nano TiO2 addition and with increasing TiO2 content.

Keywords: polycaprolactone, titanium dioxide, thermal properties, morphology

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Authors: Puchita Chokcharoenying

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Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Antioxidants are substances found in medicinal plants to help prevent heart disease, stroke, and some cancers. This study focused on evaluating the flavonoids content of Chrysanthemum Indicum and determine their antioxidant capacity by using DPPH and ABTS radical scavenging capacity assay. The total flavonoid content of C. indicumextract was determined and expressed as quercetin equivalents (QE)/g measured by an aluminiumchloride colorimetric method. The results showed that the IC50 of C. indicum extract were 83.57μg/mL ± 0.875 and52.57μg/mL ± 0.632for DPPH and ABTS, respectively. C. indicumextract exhibited antioxidant activities as a concentration dependent manner. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In summary, C. indicum extract is rich in flavonoids, which have potent antioxidant properties. Thus, C. indicum extract is a good source of antioxidants and can be developed for medicinal purposes. Nevertheless, more research on the antioxidant activity of C. indicum extract and in vivo antioxidant studies are still needed.

Keywords: ABTS assay, antioxidant, chrysanthemum indicum, DPPH assay, total flavonoid content

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Authors: Ajay Pratap Singh Lodhi, Deepak Kumar

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Vegetable oil-based environmentally friendly metalworking fluids (MWFs) are formulated. The tribological performance, cytotoxicity, and corrosion resistance of the formulated fluids (FFs) are evaluated and benchmarked with commercial mineral oil-based MWFs (CF). Results show that FFs exhibited better machining characteristics (roughness, cutting forces, and surface morphology) during machining than CF. MTT assay and Live dead cell assay confirm the cytocompatibility nature of the FFs relative to the toxic CF. Electrochemical analysis shows that FFs and CF exhibited comparable corrosion current density.

Keywords: corrosion inhibitors, cytotoxicity, machining, MTT assay, Taguchi method, vegetable oil

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Authors: Marcel Ciesla, Victoria Oberascher, Sven Eder, Stefan Kirchweger, Wolfgang E. Baaske, Gerald Ostermayer

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The omnipresent trend towards sustainable mobility is a major challenge, especially for commuters in rural areas. The use of micro public transport systems is expected to significantly reduce pollutant emissions, as several commuters travel the first mile together with a single pick-up bus instead of their own car. In this paper, different aspects of such a micro public transport system are analyzed. The main findings of the investigations should be how the travel times of commuters change and how many CO₂ emissions can be saved if some of the commuters use public transport instead of their own vehicle.

Keywords: micro public transport, green transportation, sustainable mobility, low-emission commuting

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Authors: H. Unal, A. Mimaroglu, I. Ozsoy

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In this study, the mechanical properties of micro filled epoxy composites were investigated. The matrix material is epoxy. Micro fillers are Al2O3 and TiO2 added in 10-30 wt% by weight ratio. Test samples were prepared using an open mould type die. Tensile, three point bending and hardness tests were carried out. The tensile strength, elastic modulus, elongation at break, flexural strength, flexural modulus and the hardness of the composite materials were obtained and evaluated. It was seen from the results that the level of the mechanical properties of the epoxy composites is highly influenced by micro filler content.

Keywords: composites, epoxy, fillers, mechanical properties

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Authors: Dawit Gebreegzabher Hagos, Yazezew Kebede Kiro, Mahmud Abdulkader, Henk H. D. F. Schallig, Dawit Wolday

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Rapid and accurate visceral leishmaniasis (VL) diagnosis is needed to initiate prompt treatment to reduce morbidity and mortality. Here, we evaluated the performance of loop-mediated isothermal amplification (LAMP) assay for the diagnosis of VL from blood in an endemic area in Ethiopia. LAMP was positive in 117/122 confirmed VL cases and negative in 149/152 controls, resulting in a sensitivity of 95.9% (95% CI: 90.69–98.66) and a specificity of 98.0% (95% CI: 94.34–99.59), respectively. The sensitivity of the LAMP assay was 95.0% (95% CI: 88.61–98.34) in HIV-negatives and 100% (95% CI: 85.18–100.0) in HIV-positives. Compared with microscopy, LAMP detected 82/87 (94.3%, 95% CI: 87.10–98.11) of the microscopy1 cases and was negative in 11/27 (40.7%, 95% CI: 22.39–61.20) of the microscopy2 cases. Compared with the rK39 serology, LAMP detected 113/120 (94.2%, 95% CI: 88.35–97.62) of the rK391 cases and was negative in 149/154 (96.8%, 95% CI: 92.59–98.94) of the rK392 cases. However, when compared with microscopy only, rK39 detected 83/87 (95.4%, 95% CI: 88.64–98.73) of the microscopy1 cases and negative in only 12/27 (44.4%, 95% CI: 25.48–64.67) of the microscopy– cases. There was an excellent agreement between rK39 and LAMP (Kappa 5 0.91, 95% CI: 0.86–0.96). Furthermore, an algorithm using rK39 followed by LAMP would yield a sensitivity of 99.2% (95%CI: 95.52–99.89) and a specificity of 98.0% (95% CI: 94.34–99.59). The findings demonstrate that the LAMP assay is an accurate and rapid molecular assay for VL diagnosis, including in HIV-1 co-infected patients, in an endemic setting.

Keywords: visceral leishmaniasis, HIV, diagnosis, LAMP, Ethiopia

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Authors: Mrinalini Amritkar, Disha Patil, Swapna Kulkarni, Sukratu Barve, Suresh Gosavi

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Micro-mixers play an important role in the lab-on-a-chip applications and micro total analysis systems to acquire the correct level of mixing for any given process. The mixing process can be classified as active or passive according to the use of external energy. Literature of microfluidics reports that most of the work is done on the models of steady laminar flow; however, the study of unsteady laminar flow is an active area of research at present. There are wide applications of this, out of which, we consider nanoparticle synthesis in micro-mixers. In this work, we have developed a model for unsteady flow to study the mixing performance of a passive micro mixer for reactants used for such synthesis. The model is developed in Finite Volume Method (FVM)-based software, OpenFOAM. The model is tested by carrying out the simulations at Re of 0.5. Mixing performance of the micro-mixer is investigated using simulated concentration values of mixed species across the width of the micro-mixer and calculating the variance across a line profile. Experimental validation is done by passing dyes through a Y shape micro-mixer fabricated using polydimethylsiloxane (PDMS) polymer and comparing variances with the simulated ones. Gold nanoparticles are later synthesized through the micro-mixer and collected at two different times leading to significantly different size distributions. These times match with the time scales over which reactant concentrations vary as obtained from simulations. Our simulations could thus be used to create design aids for passive micro-mixers used in nanoparticle synthesis.

Keywords: Lab-on-chip, LOC, micro-mixer, OpenFOAM, PDMS

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Authors: Kanyarat Sikhao, Nichakorn Khondee

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Adjuvant is commonly mixed with agricultural spray solution during foliar application to improve the performance of microbial-based biological control, including better spreading, absorption, and penetration on a plant leaf. This research aims to replace chemical surfactants in adjuvant by biosurfactants for reducing a negative impact on antagonistic microorganisms and crops. Biosurfactant was produced from Brevibacterium casei NK8 and used as a cell-free broth solution containing a biosurfactant concentration of 3.7 g/L. The studies of microemulsion formation and phase behavior were applied to obtain the suitable composition of biosurfactant-based adjuvant, consisting of cell-free broth (70-80%), coconut oil-based fatty alcohol C12-14 (3) ethoxylate (1-7%), and sodium chloride (8-30%). The suitable formula, achieving Winsor Type III microemulsion (bicontinuous), was 80% of cell-free broth, 7% of fatty alcohol C12-14 (3) ethoxylate, and 8% sodium chloride. This formula reduced the contact angle of water on parafilm from 70 to 31 degrees. The non-phytotoxicity against plant seed of Oryza sativa and Brassica rapa subsp. pekinensis were obtained from biosurfactant-based adjuvant (germination index equal and above 80%), while sodium dodecyl sulfate and tween80 showed phytotoxic effects to these plant seeds. The survival of Bacillus subtilis in biosurfactant-based adjuvant was higher than sodium dodecyl sulfate and tween80. The mixing of biosurfactant and plant-based surfactant could be considered as a viable, safer, and acceptable alternative to chemical adjuvant for sustainable organic farming.

Keywords: biosurfactant, microemulsion, bio-adjuvant, antagonistic microorganisms

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Authors: Rumesh Liyanage, S. B. Nawaratne, K. K. D. S. Ranaweera, Indira Wickramasinghe, K. G. S. C. Katukurunda

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Study it was attempted to identify an effect of food habits and publics’ attitudes on micro-sleepiness and preventive measures to develop a food product to combat. Statistical data pertaining to road accidents were collected from, Sri Lanka Police Traffic Division and a pre-tested questionnaire was used to collect data from 250 respondents. They were selected representing drivers (especially highway drivers), private and public sector workers (shift based) and cramming students (university and school). Questionnaires were directed to fill independently and personally and collected data were analyzed statistically. Results revealed that 76.84, 96.39 and 80.93% out of total respondents consumed rice for all three meals which lead to ingesting higher glycemic meals. Taking two hyper glycemic meals before 14.00h was identified as a cause of micro-sleepiness within these respondents. Peak level of road accidents were observed at 14.00 - 20.00h (38.2%)and intensity of micro-sleepiness falls at the same time period (37.36%) while 14.00 to 16.00h was the peak time, 16.00 to 18.00h was the least; again 18.00 to 20.00h it reappears slightly. Even though respondents of the survey expressed that peak hours of micro- sleepiness is 14.00-16.00h, according to police reports, peak hours fall in between 18.00-20.00h. Out of the interviewees, 69.27% strongly wanted to avoid micro-sleepiness and intend to spend LKR 10-20 on a commercial product to combat micro sleepiness. As age-old practices to suppress micro-sleepiness are time taken, modern day respondents (51.64%) like to have a quick solution through a drink. Therefore, food habits of morning and noon may cause for micro- sleepiness while dinner may cause for both, natural and micro-sleepiness due to the heavy glycemic load of food. According to the study micro-sleepiness, can be categorized into three zones such as low-risk zone (08.00-10.00h and 18.00-20.00h), manageable zone (10.00-12.00h), and high- risk zone (14.00-16.00h).

Keywords: food habits, glycemic load, micro-sleepiness, road accidents

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Authors: Jirathan Pongchababnapa

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Polyphenols are the most common antioxidant found in plants and are efficient in capturing oxidative free radicals. Antioxidants are substances found in medicinal plants which may have a protective role to play in certain conditions such as heart disease, stroke and some cancers. By relying on these benefits, we have traced out the presence of antioxidant in Camellia sinensis leaves extract. This study aims to evaluate flavonoids content in C. sinensisextract and investigate antioxidant activities by using DPPH and ABTS radical scavenging capacity assay. The total flavonoid content of C. Sinensis extract was determined and expressed as quercetin equivalents (QE)/g measured by the aluminum chloride colorimetric method. The results showed that the IC₅₀ of C. Sinensis leaves extract were 40.90 μg/mL ± 0.755 and32.96 μg/mL ± 0.679 for DPPH and ABTS, respectively. C. Sinensis extract at increasing concentration showed antioxidant activities as a concentration dependent manner. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. Sinensis extract consisted of a high amount of flavonoids content which possesses potent antioxidant activity. However, further investigation on the identification of pure compound of this plant and molecular antioxidant assays are still required.

Keywords: ABTS assay, antioxidant, camellia sinensis, DPPH assay, total flavonoid content

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Authors: S. U. Khan, T. Ayub, N. Shafiq

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The properties of locally produced metakaolin (MK) as cement replacing material and the comparison of reactivity with commercially available micro-silica have been investigated. Compressive strength, splitting tensile strength, and load-deflection behaviour under bending are the properties that have been studied. The amorphous phase of MK with micro-silica was compared through X-ray diffraction (XRD) pattern. Further, interfacial transition zone of concrete with micro-silica and MK was observed through Field Emission Scanning Electron Microscopy (FESEM). Three mixes of concrete were prepared. One of the mix is without cement replacement as control mix, and the remaining two mixes are 10% cement replacement with micro-silica and MK. It has been found that MK, due to its irregular structure and amorphous phase, has high reactivity with portlandite in concrete. The compressive strength at early age is higher with MK as compared to micro-silica. MK concrete showed higher splitting tensile strength and higher load carrying capacity as compared to control and micro-silica concrete at all ages respectively.

Keywords: metakaolin, compressive strength, splitting tensile strength, load deflection, interfacial transition zone

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Authors: Amir Davood Elmi

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From the earliest light microscope to the most innovative X-ray imaging techniques, all of them have refined and improved our knowledge about the organization and composition of living tissues. The old techniques are time consuming and ultimately destructive to the tissues under the examination. In recent few decades, thanks to the boost of technology, non-destructive visualization techniques, such as X-ray computed tomography (CT), magnetic resonance imaging (MRI), selective plane illumination microscopy (SPIM), and optical projection tomography (OPT), have come to the forefront. Among these techniques, CT is excellent for mineralized tissues such as bone or dentine. In addition, CT it is faster than other aforementioned techniques and the sample remains intact. In this article, applications, advantages, and limitations of micro-CT is discussed, in addition to some information about micro-CT of soft tissue.

Keywords: Micro-CT, hard tissue, bone, attenuation coefficient, rapid prototyping

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Authors: K. Kefif, Y. Bouizem, A. Belfedal, D. J. Sib, K. Zellama, l. Chahed

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Hydrogenated microcrystalline silicon (μc-Si:H) thin films were prepared by radio frequency magnetron sputtering at relatively low growth temperatures (Ts=100 °C). The films grown on glass substrate in order to use the new generation of substrates sensitive to elevated temperatures. Raman spectroscopy was applied to investigate the effect of the argon gas diluted in hydrogen, on the structural properties and the evolution of the micro structure in the films. Raman peak position, intensity and line width were used to characterize the quality and the percentage of the crystallites in the films. The results of this investigation suggest the existence of a threshold dilution around a gas mixture of argon (40%) and hydrogen (60%) for which the crystallization occurs, even at low deposition temperatures. The difference between the amorphous and the crystallized structures is well confirmed by spectroscopic ellipsometry (SE) technique.

Keywords: Silicon, Thin films, Structural properties, Raman spectroscopy, Ellipsometry

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Authors: Tawqeer Ali Syed, Prakash Chandra

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This study aims to evaluate the antileishmanial activity of Nigella sativa black seed extract. This well-known plant extract was taken from the botanical garden of Kashmir. Materials and Methods: The methanolic extracts of these plants were screened for their antileishmanial activity against Leishmania major using 3‑(4.5‑dimethylthiazol‑2yl)‑2.5‑diphenyltetrazolium bromide assay or MTT assay. Results: The methanolic extract of Nigella sativa showed potential antileishmanial activity at an inhibition% value of 80.29% ± 0.65%. IC 50 was calculated after 48 hours to be 964.3 µg/ml. Conclusion: Considering these results, these medicinal plants from Kashmir could serve as potential drug sources for antileishmanial compounds.

Keywords: MTT assay, antileishmanial, cell viability, Nigella sativa

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Authors: Lozano Paul

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Coupled Membrane Separation Technology (CMST), including Cross Flow Microfiltration (CFM) and Reverse Osmosis (RO), are used to concentrate microalgae biomass or/and to extract and concentrate water-soluble metabolites produced during micro-algae production cycle, as well as water recycling. Micro-algae biomass was produced using different feeding mixtures of ingredients: pure chemical origin compounds and natural/ecological water-extracted components from available local plants. Micro-algae was grown either in conventional plastic bags (100L/unit) or in small-scale innovative bioreactors (75L). Biomass was concentrated as CFM retentate using a P19-60 ceramic membrane (0.2μm pore size), and water-soluble micro-algae metabolites left in the CFM filtrate were concentrated by RO. Large volumes of water (micro-algae culture media) of were recycled by the CMTS for another biomass production cycle.

Keywords: extraction, membrane process, microalgae, natural compound

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Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn

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Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.

Keywords: cellulose, fermentation, grammage, paper, Spirogyra sp.

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Authors: Monthon Tangjitmungman

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Numerous diseases have been linked to oxidative stress, in which a disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Due to the presence of phenolic compounds in Caesalpinia sappan has been discovered to have antioxidant activity. It has several health benefits, the most important of which is preventing cardiovascular and cancer diseases. This study aimed to determine the phenolic content and antioxidant activity of C. sappan extract using a variety of antioxidant assays. The extract of C. sappan was made using a mixture of solvents (ethyl alcohol: water in ratio 8:2). The total phenolic content of C. sappan extract was determined and expressed as gallic acid equivalents using the Folin-Cioucalteu method (GAE). The antioxidant activity of C. sappan extract was assessed using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and the ABTS radical scavenging capacity assay. An association was found between antioxidant activity and total phenol content. The antioxidant activity of C. sappan extract was also determined by DPPH and ABTS assays. The IC50 values for C. sappan extract from DPPH and ABTS assays were 54.48 μg/mL ± 0.545 and 25.46 μg/mL ± 0.790, respectively, in the DPPH assay. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. sappan extract contains a high level of total phenolics and exhibits significant antioxidant activity. Nevertheless, more research should be done on the antioxidant activity, such as SOD and ROS scavenging assays and in vivo experiments, to determine whether the compound has antioxidant activity.

Keywords: ABTS assay, antioxidant activity, Caesalpinia sappan, DPPH assays, total phenolic content

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Authors: Thilivhali Emmanuel Tshikalange, Darky Cheron Modishane, Frederick Tawi Tabit

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The fruit pulp extracts of twelve selected ethnobotanical wild edible fruits from Mutale local municipality in Venda (Limpopo Province, South Africa) were investigated for their antimicrobial, antioxidant and cytotoxicity activities. Methanol extracts were prepared and tested against six micro-organisms (Salmonella typhi, Streptococcus pyogenes, Bacillus cereus, Klebsiella pneumoniae, Prevotella intermedia and Candida albicans). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the micro-dilution method, while for antioxidant activity the 2,2-diphenyl-1-picrylhydrazyl method was used. Of the 12 extracts tested, Adonsonia digitata, Berchemia discolor, Manilkara mochisia, Xanthocercis zambesiaca, Landolphia kirkii and Garcinia livingstonei showed antimicrobial activity, with MIC values ranging from 12.5 to 0.4 mg/ml. Gram negative bacteria were more resistant to the extracts in comparison to Gram positive bacteria. Antioxidant activity was only detected in Adonsonia digitata extract and the IC50 (substrate concentration to produce 50% reduction) was found to be 16.18µg/ml. The cytotoxicity of the extracts that showed antimicrobial and antioxidant activities was also determined. All plant extracts tested were non-toxic against human kidney cells (HEK293), with IC50 values of >400 µg/ml. The results presented in this study provide support to some traditional uses of wild edible fruits.

Keywords: antimicrobial, antioxidant, cytotoxicity, ethnobotanical, fruits

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Authors: Ibrahim Aly, Rabab Zalat, Bahaa EL Deen W. El Aswad, Ismail M. Moharm, Basam M. Masoud, Tarek Diab

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The objective of the present study was to evaluate the novel nanomagnetic beads based–latex agglutination assay (NMB-LAT) as a simple test for diagnosis of S. haematobium as well as standardize the novel nanomagnetic beads based –ELISA (NMB-ELISA). According to urine examination this study included 85 S. haematobium infected patients, 30 other parasites infected patients and 25 negative control samples. The sensitivity of novel NMB-LAT was 82.4% versus 96.5% and 88.2% for NMB-ELISA and currently used sandwich ELISA respectively. The specificity of NMB-LAT was 83.6% versus 96.3% and 87.3% for NMB-ELISA and currently used sandwich ELISA respectively. In conclusion, the novel NMB-ELISA is a valuable applicable diagnostic technique for diagnosis of human schistosomiasis haematobium. The novel NMB-ELISA assay is a suitable applicable diagnostic method in field survey especially when followed by ELISA as a confirmatory test in query false negative results. Trials are required to increase the sensitivity and specificity of NMB-ELISA assay.

Keywords: diagnosis, iatex agglutination, nanomagnetic beads, sandwich ELISA

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Authors: Shayma Munqith Al-Baker, Fadhl Ahmed Saeed Al-Gasha’a, Samira Hamid Hanash, Ahmed Ali Al-Hazmi

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A total of 200 samples (180 fecal materials and 20 organ samples) were collected from (5 different poultry farms, 10 local poultry shops, 5 houses poultry, 5 Eggs stores shops and 5 hand slaughters centers) in Ibb city, Yemen, 2014. According to morphological, cultural, as well as biochemical characterization and serological tests, 59 29.5% isolates were identified as Salmonella spp. and all Salmonella isolates were categorized by serotype, which comprised of, 37 62.71% Salmonella Typhimurium serovar, 21 35.59%. Salmonella Enteritidis serovar and 11.69% Salmonella Heidelberg serovar. Antibiotic sensitivity test was done for bacterial isolates and the results showed there were clear differences in antibiotic resistant. Antimicrobial susceptibility of the isolates varies as follows: Ofloxacin 79.66%, Ciprofloxacin 67.80%, Colistin 59.32% and Gentamycin 52.54%. All of isolates were resistant to Erythromycin, Penicillin and Lincomycin. Antibacterial activity was done for both aqueous and ethanol extracts of Dodonaea viscosa plant by using well and disc diffusion assay. The results indicated that well diffusion assay had best results than disc diffusion assay, the highest inhibition zone was 22 mm for well diffusion and 15 mm for disc diffusion assay, the results observed that ethanol extract had best antibacterial effect than aqueous extract which the percentage of bacterial isolates affected with ethanol extract was 71.19% comparing with aqueous extract 28.81% by using disc diffusion assay, while the percentage of bacterial isolates affected with ethanol extract was 88.13% comparing with aqueous extract 52.54% by using will diffusion assay.

Keywords: Salmonella spp, Dodonaea viscosa, antimicrobial and salmonellosis

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Authors: Ayman K. El Essawy, Amal M. Hosny, Hala M. Abu Shady

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The rapid detection of TB and drug resistance, both optimizes treatment and improves outcomes. In the current study, respiratory specimens were collected from 155 patients. Conventional susceptibility testing and MIC determination were performed for rifampicin (RIF) and isoniazid (INH). Genotype MTBDRplus assay, which is a molecular genetic assay based on the DNA-STRIP technology and specific gene sequencing with primers for rpoB, KatG, and mab-inhA genes were used to detect mutations associated with resistance to rifampicin and isoniazid. In comparison to other categories, most of rifampicin resistant (61.5%) and isoniazid resistant isolates (47.1%) were from patients relapsed in treatment. The genotypic profile (using Genotype MTBDRplus assay) of multi-drug resistant (MDR) isolates showed missing of katG wild type 1 (WT1) band and appearance of mutation band katG MUT2. For isoniazid mono-resistant isolates, 80% showed katG MUT1, 20% showed katG MUT1, and inhA MUT1, 20% showed only inhA MUT1. Accordingly, 100% of isoniazid resistant strains were detected by this assay. Out of 17 resistant strains, 16 had mutation bands for katG distinguished high resistance to isoniazid. The assay could clearly detect rifampicin resistance among 66.7% of MDR isolates that showed mutation band rpoB MUT3 while 33.3% of them were considered as unknown. One mono-resistant rifampicin isolate did not show rifampicin mutation bands by Genotype MTBDRplus assay, but it showed an unexpected mutation in Codon 531 of rpoB by DNA sequence analysis. Rifampicin resistance in this strain could be associated with a mutation in codon 531 of rpoB (based on molecular sequencing), and Genotype MTBDRplus assay could not detect the associated mutation. If the results of Genotype MTBDRplus assay and sequencing were combined, this strain shows hetero-resistance pattern. Gene sequencing of eight selected isolates, previously tested by Genotype MTBDRplus assay, could detect resistance mutations mainly in codon 315 (katG gene), position -15 in inhA promotes gene for isoniazid resistance and codon 531 (rpoB gene) for rifampicin resistance. Genotyping techniques allow distinguishing between recurrent cases of reinfection or reactivation and supports epidemiological studies.

Keywords: M. tuberculosis, rpoB, KatG, inhA, genotype MTBDRplus

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Authors: Aqeela Ashraf, Muhammad Imran, Tahir Yaqub, Muhammad Tayyab, Yung Fu Chang

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For the identification of bovine mastitic pathogen, an economical, rapid and sensitive molecular diagnostic assay is developed by PCR multiplexing of gene and pathogenic species specific DNA sequences. The multiplex PCR assay is developed for detecting nine important bacterial pathogens causing mastitis Worldwide. The bacterial species selected for this study are Streptococcus agalactiae, Streptococcus dysagalactiae, Streptococcus uberis, Staphylococcus aureus, Escherichia coli, Staphylococcus haemolyticus, Staphylococcus chromogenes Mycoplasma bovis and Staphylococcus epidermidis. A single reaction assay was developed and validated by 27 reference strains and further tested on 276 bacterial strains obtained from culturing mastitic milk. The multiplex PCR assay developed here is further evaluated by applying directly on genomic DNA isolated from 200 mastitic milk samples. It is compared with bacterial culturing method and proved to be more sensitive, rapid, economical and can specifically identify 9 bacterial pathogens in a single reaction. It has detected the pathogens in few culture negative mastitic samples. Recognition of disease is the foundation of disease control and prevention. This assay can be very helpful for maintaining the udder health and milk monitoring.

Keywords: multiplex PCR, bacteria, mastitis, milk

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Authors: Shofiq Ahmed, Rakibul Hassan, Raquib Ahsan

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Concrete is one of the basic elements of RCC structure and also the most crucial one. In recent years, a lot of researches have been conducted to develop special types of concrete for special purposes. Micro-concrete is one of them which has high compressive strength and is mainly used for retrofitting. Micro-concrete is a cementitious based composition formulated for use in repairs of areas where the concrete is damaged & the area is confined in movement making the placement of conventional concrete difficult. According to recent statistics, a large number of structures in the major cities of Bangladesh are vulnerable to collapse. Retrofitting may thus be required for a sustainable solution, and for this purpose, the utilization of micro-concrete can be considered as the most effective solution. For that reason, the aim of this study was to produce micro-concrete using indigenous materials in low cost. Following this aim, the experimental data were observed for five mix ratios with varied amount of cement, fine aggregate, coarse aggregate, water, and admixture. The investigation criteria were a compressive strength, tensile strength, slump and the cost of different mix ratios. Finally, for a mix ratio of 1:1:1.5, the compressive strength was achieved as 7820 psi indicating highest strength among all the samples with the reasonable tensile strength of 1215 psi. The slump of 6.9 inches was also found for this specimen indicating it’s high flowability and making it’s convenient to use as micro-concrete. Moreover, comparing with the cost of foreign products of micro-concrete, it was observed that foreign products were almost four to five times costlier than this local product.

Keywords: indigenous, micro-concrete, retrofitting, vulnerable

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Authors: Julie Connelly, Tanvi Toprani, Xin Xie, Dhinoth Kumar Bangarusamy, Kris C. Gunsalus

Abstract:

The overuse of antibiotics worldwide has contributed to the development of multi-drug resistant (MDR) pathogenic bacterial strains. There is an increasing urgency to discover antibiotics to combat MDR pathogens. The microbiome of the Arabian Gulf is a largely unexplored and potentially rich source of novel bioactive compounds. Microbes that inhabit the Abu Dhabi coastal regions adapt to extreme environments with high salinity, hot temperatures, large temperature fluctuations, and acute exposure to solar energy. The microbes native to this region may produce unique metabolites with therapeutic potential as antibiotics and antifungals. We have isolated 200 pure bacterial strains from mangrove sediments, cyanobacterial mats, and coral reefs of the Abu Dhabi region. In this project, we aim to screen the marine bacterial strains to identify antibiotics, in particular undocumented compounds that show activity against existing antibiotic-resistant strains. We have acquired the ESKAPE pathogen panel, which consists of six antibiotic-resistant gram-positive and gram-negative bacterial pathogens that collectively cause most clinical infections. Our initial efforts of the primary screen using colony-picking co-culture assay have identified several candidate marine strains producing potential antibiotic compounds. We will next apply different assays, including disk-diffusion and broth turbidity growth assay, to confirm the results. This will be followed by bioactivity-guided purification and characterization of target compounds from the scaled-up volume of candidate strains, including SPE fraction, HPLC fraction, LC-MS, and NMR. For antimicrobial compounds with unknown structures, our final goal is to investigate their mode of action by identifying the molecular target.

Keywords: marine bacteria, natural products, drug discovery, ESKAPE panel

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Authors: J. Hidalgo de Quintana, I. Stoner, M. Tackett, G. Doran, C. Rafferty, A. Windemuth, J. Tytell, D. Pregibon

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We have developed the Multiplexed Circulating microRNA assay that allows the detection of up to 68 microRNA targets per sample. The assay combines particle­based multiplexing, using patented Firefly hydrogel particles, with single­ step RT-PCR signal. Thus, the Circulating microRNA assay leverages PCR sensitivity while eliminating the need for separate reverse transcription reactions and mitigating amplification biases introduced by target­-specific qPCR. Furthermore, the ability to multiplex targets in each well eliminates the need to split valuable samples into multiple reactions. Results from the Circulating microRNA assay are interpreted using Firefly Analysis Workbench, which allows visualization, normalization, and export of experimental data. To aid discovery and validation of biomarkers, we have generated fixed panels for Oncology, Cardiology, Neurology, Immunology, and Liver Toxicology. Here we present the data from several studies investigating circulating and tumor microRNA, showcasing the ability of the technology to sensitively and specifically detect microRNA biomarker signatures from fluid specimens.

Keywords: biomarkers, biofluids, miRNA, photolithography, flowcytometry

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Authors: Prachi Choudhary, Jai Gopal Sharma

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Microbial cultures determine the type of organism, its abundance in the tested sample, or both. It is one of the primary diagnostic methods of microbiology. It is used to determine the cause of infectious disease by letting the agent multiply in a predetermined medium. Different bacteria produce colonies that may be very distinct from the bacterial species that produced them. To culture any pathogen or microorganism, we should first know about the types of media used in microbiology for culturing. Sometimes sub culturing is also done in various microorganisms if some mixed growth is seen in culture. Nearly 3 types of culture media based on consistency – solid, semi-solid, and liquid (broth) media; are further explained in the report. Then, The Five I's approach is a method for locating, growing, observing, and characterizing microorganisms, including inoculation and incubation. Isolation, inspection, and identification. For identification of bacteria, we have to culture the sample like urine, sputum, blood, etc., on suitable media; there are different methods of culturing the bacteria or microbe like pour plate method, streak plate method, swabbing by needle, pipetting, inoculation by loop, spreading by spreader, etc. After this, we see the bacterial growth after incubation of 24 hours, then according to the growth of bacteria antibiotics susceptibility test is conducted; this is done for sensitive antibiotics or resistance to that bacteria, and also for knowing the name of bacteria. Various methods like the dilution method, disk diffusion method, E test, etc., do antibiotics susceptibility tests. After that, various medicines are provided to the patients according to antibiotic sensitivity and resistance.

Keywords: inoculation, incubation, isolation, antibiotics suspectibility test, characterizing

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Authors: Abdul Matin, Muazzama Akhtar, Shahid Nisar, Saddaf Mazzar, Umer Rashid

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Antibiotic resistance is an increasing concern worldwide now a day. Neisseria gonorrhea and Staphylococcus aureus are known to cause major human sexually transmitted and respiratory diseases respectively. Nanotechnology is an emerging discipline and its application in various fields especially in medical sciences is gigantic. In the present study, we synthesized multi-walled carbon nanotubes (MWNTs) using acid oxidation method and solubilized MWNTs were with length predominantly >500 nm and diameters ranging from 40 to 50 nm. The locally synthesized MWNTs were used against gram positive and negative bacteria to determine their impact on bacterial growth. Clinical isolates of Neisseria gonorrhea (isolate: 4C-11) and Staphylococcus aureus (isolate: 38541) were obtained from local hospital and normally cultured in LB broth at 37°C. Both clinical strains can be obtained on request from University of Gujarat. Spectophometric assay was performed to determine the impact of MWNTs on bacterial growth in vitro. To determine the effect of MWTNs on test organisms, various concentration of MWNTs were used and recorded observation on various time intervals to understand the growth inhibition pattern. Our results demonstrated that MWNTs exhibited toxic effects to Staphylococcus aureus while showed very limited growth inhibition to Neisseria gonorrhea, which suggests the resistant potential of Neisseria against nanoparticles. Our results clearly demonstrate the gradual decrease in bacterial numbers with passage of time when compared with control. Maximum bacterial inhibition was observed at maximum concentration (50 µg/ml). Our future work will include further characterization and mode of action of our locally synthesized MWNTs. In conclusion, we investigated and reported for the first time the inhibitory potential of locally synthesized MWNTs on local clinical isolates of Staphylococcus aureus and Neisseria gonorrhea.

Keywords: antibacterial activity, multi walled carbon nanotubes, Neisseria gonorrhea, spectrophotometer assay, Staphylococcus aureus

Procedia PDF Downloads 286