Search results for: organic sulfur bacteria
1986 Antimicrobial Activities of Lactic Acid Bacteria from Fermented Foods and Probiotic Products
Authors: Alec Chabwinja, Cannan Tawonezvi, Jerneja Vidmar, Constance Chingwaru, Walter Chingwaru
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Objective: To evaluate the potential of commercial fermented / probiotic products available in Zimbabwe or internationally, and strains of Lactobacillus plantarum (L. plantarum) as prophylaxis and therapy against diarrhoeal and sexually transmitted infections. Methods: The antimicrobial potential of cultures of lactobacilli enriched from 4 Zimbabwean commercial food/beverage products, namely Dairibord Lacto sour milk (DLSM), Probrand sour milk (PSM), Kefalos Vuka cheese (KVC) and Chibuku opaque beer (COB); three probiotic products obtainable in Europe and internationally; and four strains of L. plantarum obtained from Balkan traditional cheeses and Zimbabwean foods against clinical strains of Escherichia coli (E. coli) and non-clinical strains of Candida albicans and Rhodotorula spp. was assayed using the well diffusion method. Three commercial Agar diffusion assay and a competitive exclusion assay were carried out on Mueller-Hinton agar. Results: Crude cultures of putative lactobacillus strains obtained from Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer) exhibited significantly greater antimicrobial activities against clinical strains of E. coli than strains of L. plantarum isolated from Balkan cheeses (CLP1, CLP2 or CLP3) or crude microbial cultures from commercial paediatric probiotic products (BG, PJ and PL) of a culture of Lactobacillus rhamnosus LGG (p < 0.05). Furthermore, the following has high antifungal activities against the two yeasts: supernatant-free microbial pellet (SFMP) from an extract of M. azedarach leaves (27mm ± 2.5) > cell-free culture supernatants (CFCS) from Maaz Dairy sour milk and Mnandi sour milk (approximately 26mm ± 1.8) > CFCS and SFMP from Amansi hodzeko (25mm ± 1.5) > CFCS from Parinari curatellifolia fruit (24mm ± 1.5), SFMP from P. curatellifolia fruit (24mm ± 1.4) and SFMP from mahewu (20mm ± 1.5). These cultures also showed high tolerance to acidic conditions (~pH4). Conclusions: The putative lactobacilli from four commercial Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer), and three strains of L. plantarum from Balkan cheeses (CLP1, CLP2 or CLP3) exhibited high antibacterial activities, while Maaz Dairy sour-, Mnandi sour- and Amansi hodzeko milk products had high antifungal activities. Our selection of Zimbabwean probiotic products has potential for further development into probiotic products for use in the control diarrhea caused by pathogenic strains of E. coli or yeast infections. Studies to characterise the probiotic potential of the live cultures in the products are underway.Keywords: lactic acid bacteria, Staphylococcus aureus, Streptococcus spp, yeast, inhibition, acid tolerance
Procedia PDF Downloads 4121985 The Antimicrobrial Effect of Alkaloids (Harmin, Harmalin) Extracted from Peganum harmala (L) Seeds in the South of Algeria (Bousaada)
Authors: Nassima Behidj-Benyounes, Thoraya Dahmene, Nadjiba Chebout
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This work examines the study of the antimicrobrial effect of alkaloids extracted from the seeds of Peganum harmala L (Zygophyllaceae). This natural substance is extracted by using different solvents (aqueous, ethanolic, and hexane). The evaluation of the antimicrobial activity has only dealt with alkaloids. The antimicrobial effect of alkaloids is evaluated on several microorganisms. It has been tested on eight bacterial strains. The extract has been studied by using two yeasts. Finally, three molds have been studied. It should be noted that these agents are characterized by a high frequency of contamination and pathogenicity. Through this study, we note that Staphylococcus aureus, Saccharomyces cerievisae and E. coli are very sensitive in respect of the ethanol extract. Pseudomonas aerogenosa and Penicillium sp. are resistant to this extract. The other microorganisms are moderately sensitive. The study of the antimicrobial activity of different extracts of the Harmel has shown an optimal activity with the ethanol extract.Keywords: Peganum harmala L., seeds, alkaloids, bacteria, fungi, yeast, antimicrobial activity
Procedia PDF Downloads 4031984 Synthesis and Characterization of Zinc (II) Complex and Its Catalytic Activity on C(SP3)-H Oxidation Reactions
Authors: Yalçın Kılıç, İbrahim Kani
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The conversion of hydrocarbons to carbonyl compounds by oxidation reaction is one of the most important reactions in the synthesis of fine chemicals. As a result of the oxidation of hydrocarbons containing aliphatic sp3-CH groups in their structures, aldehydes, ketones or carboxylic acids can be obtained. In this study, OSSO-type 2,2'-[1,4-butanedylbis(thio)]bis-benzoic acid (tsabutH2) ligand and [Zn(µ-tsabut)(phen)]n complex (where phen = 1,10-phenantroline) were synthesized and their structures were characterized by single crystal x-ray diffraction method. The catalytic efficiency of the complex in the catalytic oxidation studies of organic compounds such as cyclohexane, ethylbenzene, diphenylmethane, and p-xylene containing sp3-C-H in its structure was investigated.Keywords: metal complex, OSSO-type ligand, catalysis, oxidation
Procedia PDF Downloads 1011983 Activity of Malate Dehydrogenase in Cell Free Extracts from S. proteamaculans, A. hydrophila, and K. pneumoniae
Authors: Mohamed M. Bumadian, D. James Gilmour
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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase
Procedia PDF Downloads 4651982 A Universal Hybrid Adsorbent Based on Chitosan for Water Treatment
Authors: Sandrine Delpeux-Ouldriane, Min Cai, Laurent Duclaux, Laurence Reinert, Fabrice Muller
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A novel hybrid adsorbent, based on chitosan biopolymer, clays and activated carbon was prepared. Hybrid chitosan beads containing dispersed clays and activated carbons were prepared by precipitation in basic medium. Such a composite material is still very porous and presents a wide adsorption spectrum. The obtained composite adsorbent is able to handle all the pollution types including heavy metals, polar and hydrophobic organic molecules and nitrates. It could find a place of choice in tertiary water treatment processes or for an ‘at source’ treatment concerning chemical or pharmaceutical industries.Keywords: adsorption, chitosan, clay mineral, activated carbon
Procedia PDF Downloads 4041981 Factors Controlling Marine Shale Porosity: A Case Study between Lower Cambrian and Lower Silurian of Upper Yangtze Area, South China
Authors: Xin Li, Zhenxue Jiang, Zhuo Li
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Generally, shale gas is trapped within shale systems with low porosity and ultralow permeability as free and adsorbing states. Its production is controlled by properties, in terms of occurrence phases, gas contents, and percolation characteristics. These properties are all influenced by porous features. In this paper, porosity differences of marine shales were explored between Lower Cambrian shale and Lower Silurian shale of Sichuan Basin, South China. Both the two shales were marine shales with abundant oil-prone kerogen and rich siliceous minerals. Whereas Lower Cambrian shale (3.56% Ro) possessed a higher thermal degree than that of Lower Silurian shale (2.31% Ro). Samples were measured by a combination of organic-chemistry geology measurement, organic matter (OM) isolation, X-ray diffraction (XRD), N2 adsorption, and focused ion beam milling and scanning electron microscopy (FIB-SEM). Lower Cambrian shale presented relatively low pore properties, with averaging 0.008ml/g pore volume (PV), averaging 7.99m²/g pore surface area (PSA) and averaging 5.94nm average pore diameter (APD). Lower Silurian shale showed as relatively high pore properties, with averaging 0.015ml/g PV, averaging 10.53m²/g PSA and averaging 18.60nm APD. Additionally, fractal analysis indicated that the two shales presented discrepant pore morphologies, mainly caused by differences in the combination of pore types between the two shales. More specifically, OM-hosted pores with pin-hole shape and dissolved pores with dead-end openings were the main types in Lower Cambrian shale, while OM-hosted pore with a cellular structure was the main type in Lower Silurian shale. Moreover, porous characteristics of isolated OM suggested that OM of Lower Silurian shale was more capable than that of Lower Cambrian shale in the aspect of pore contribution. PV of isolated OM in Lower Silurian shale was almost 6.6 times higher than that in Lower Cambrian shale, and PSA of isolated OM in Lower Silurian shale was almost 4.3 times higher than that in Lower Cambrian shale. However, no apparent differences existed among samples with various matrix compositions. At late diagenetic or metamorphic epoch, extensive diagenesis overprints the effects of minerals on pore properties and OM plays the dominant role in pore developments. Hence, differences of porous features between the two marine shales highlight the effect of diagenetic degree on OM-hosted pore development. Consequently, distinctive pore characteristics may be caused by the different degrees of diagenetic evolution, even with similar matrix basics.Keywords: marine shale, lower Cambrian, lower Silurian, om isolation, pore properties, om-hosted pore
Procedia PDF Downloads 1351980 Introduction of Microbial Symbiosis in Genus of Tridacna and Kiwaidae with Insights into Aquaculture
Authors: Jincao Guo
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Aquaculture plays a significant role in the diet of people in many regions. However, problems such as bioaccumulation have risen with the rapidly growing industry due to a lack of control in the feeding process, which brings uncertainty to the quality of the products. The paper tackles the problem by introducing the symbiosis of the Giant Clam (Tridacna) with photosynthetic algae and Yeti Crab (Kiwaidae) with chemosynthetic bacteria in molecular and developmental details. By combing the knowledge gained from the two models and past studies, innovative ideas such as using mass selection methods to domesticate and farm those symbiotic species, as well as improvements for the current farming methods, such as introducing algae feeding, are discussed. Further studies are needed, but experiments are worth conducting since it increases the variety of choices for consumers and can potentially improve the quality and efficiency of aquaculture.Keywords: the giant clam Tridacna, yeti crab Kiwaidae, autotroph microbes, microbial symbiosis, aquaculture, bivalves, crustaceans, mollusk, photosynthesis, chemosynthesis
Procedia PDF Downloads 781979 Green Synthesis (Using Environment Friendly Bacteria) of Silver-Nanoparticles and Their Application as Drug Delivery Agents
Authors: Sutapa Mondal Roy, Suban K. Sahoo
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The primary aim of this work is to synthesis silver nanoparticles (AgNPs) through environmentally benign routes to avoid any chemical toxicity related undesired side effects. The nanoparticles were stabilized with drug ciprofloxacin (Cp) and were studied for their effectiveness as drug delivery agent. Targeted drug delivery improves the therapeutic potential of drugs at the diseased site as well as lowers the overall dose and undesired side effects. The small size of nanoparticles greatly facilitates the transport of active agents (drugs) across biological membranes and allows them to pass through the smallest capillaries in the body that are 5-6 μm in diameter, and can minimize possible undesired side effects. AgNPs are non-toxic, inert, stable, and has a high binding capacity and thus can be considered as biomaterials. AgNPs were synthesized from the nutrient broth supernatant after the culture of environment-friendly bacteria Bacillus subtilis. The AgNPs were found to show the surface plasmon resonance (SPR) band at 425 nm. The Cp capped Ag nanoparticles formation was complete within 30 minutes, which was confirmed from absorbance spectroscopy. Physico-chemical nature of the AgNPs-Cp system was confirmed by Dynamic Light Scattering (DLS), Transmission Electron Microscopy (TEM) etc. The AgNPs-Cp system size was found to be in the range of 30-40 nm. To monitor the kinetics of drug release from the surface of nanoparticles, the release of Cp was carried out by careful dialysis keeping AgNPs-Cp system inside the dialysis bag at pH 7.4 over time. The drug release was almost complete after 30 hrs. During the drug delivery process, to understand the AgNPs-Cp system in a better way, the sincere theoretical investigation is been performed employing Density Functional Theory. Electronic charge transfer, electron density, binding energy as well as thermodynamic properties like enthalpy, entropy, Gibbs free energy etc. has been predicted. The electronic and thermodynamic properties, governed by the AgNPs-Cp interactions, indicate that the formation of AgNPs-Cp system is exothermic i.e. thermodynamically favorable process. The binding energy and charge transfer analysis implies the optimum stability of the AgNPs-Cp system. Thus, the synthesized Cp-Ag nanoparticles can be effectively used for biological purposes due to its environmentally benign routes of synthesis procedures, which is clean, biocompatible, non-toxic, safe, cost-effective, sustainable and eco-friendly. The Cp-AgNPs as biomaterials can be successfully used for drug delivery procedures due to slow release of drug from nanoparticles over a considerable period of time. The kinetics of the drug release show that this drug-nanoparticle assembly can be effectively used as potential tools for therapeutic applications. The ease of synthetic procedure, lack of possible chemical toxicity and their biological activity along with excellent application as drug delivery agent will open up vista of using nanoparticles as effective and successful drug delivery agent to be used in modern days.Keywords: silver nanoparticles, ciprofloxacin, density functional theory, drug delivery
Procedia PDF Downloads 3871978 Assessment of Bisphenol A and 17 α-Ethinyl Estradiol Bioavailability in Soils Treated with Biosolids
Authors: I. Ahumada, L. Ascar, C. Pedraza, J. Montecino
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It has been found that the addition of biosolids to soil is beneficial to soil health, enriching soil with essential nutrient elements. Although this sludge has properties that allow for the improvement of the physical features and productivity of agricultural and forest soils and the recovery of degraded soils, they also contain trace elements, organic trace and pathogens that can cause damage to the environment. The application of these biosolids to land without the total reclamation and the treated wastewater can transfer these compounds into terrestrial and aquatic environments, giving rise to potential accumulation in plants. The general aim of this study was to evaluate the bioavailability of bisphenol A (BPA), and 17 α-ethynyl estradiol (EE2) in a soil-biosolid system using wheat (Triticum aestivum) plant assays and a predictive extraction method using a solution of hydroxypropyl-β-cyclodextrin (HPCD) to determine if it is a reliable surrogate for this bioassay. Two soils were obtained from the central region of Chile (Lo Prado and Chicauma). Biosolids were obtained from a regional wastewater treatment plant. The soils were amended with biosolids at 90 Mg ha-1. Soils treated with biosolids, spiked with 10 mgkg-1 of the EE2 and 15 mgkg-1 and 30 mgkg-1of BPA were also included. The BPA, and EE2 concentration were determined in biosolids, soils and plant samples through ultrasound assisted extraction, solid phase extraction (SPE) and gas chromatography coupled to mass spectrometry determination (GC/MS). The bioavailable fraction found of each one of soils cultivated with wheat plants was compared with results obtained through a cyclodextrin biosimulator method. The total concentration found in biosolid from a treatment plant was 0.150 ± 0.064 mgkg-1 and 12.8±2.9 mgkg-1 of EE2 and BPA respectively. BPA and EE2 bioavailability is affected by the organic matter content and the physical and chemical properties of the soil. The bioavailability response of both compounds in the two soils varied with the EE2 and BPA concentration. It was observed in the case of EE2, the bioavailability in wheat plant crops contained higher concentrations in the roots than in the shoots. The concentration of EE2 increased with increasing biosolids rate. On the other hand, for BPA, a higher concentration was found in the shoot than the roots of the plants. The predictive capability the HPCD extraction was assessed using a simple linear correlation test, for both compounds in wheat plants. The correlation coefficients for the EE2 obtained from the HPCD extraction with those obtained from the wheat plants were r= 0.99 and p-value ≤ 0.05. On the other hand, in the case of BPA a correlation was not found. Therefore, the methodology was validated with respect to wheat plants bioassays, only in the EE2 case. Acknowledgments: The authors thank FONDECYT 1150502.Keywords: emerging compounds, bioavailability, biosolids, endocrine disruptors
Procedia PDF Downloads 1471977 Antibacterial and Antityrosinase Activity of Isolated Compounds from Stem Bark of Ficus platyphylla Del
Authors: Aminu Muhammad, Mustapha Ya’u, Hasnah Mohd Sirat
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An investigation of the chemical constituents into the stem bark of Ficus platyphylla (Moraceae) has resulted in the isolation of hordenine, epicatechin, lupeol, lupeol acetate and α-amyrin acetate. Their structures were determined using spectroscopic data as well as comparison with literature data. The antibacterial assay has been tested against Gram positive and Gram negative bacteria, while the tyrosinase inhibition assay was examined using L-Dopa as a substrate of mushroom tyrosinase enzyme. hordenine, epicatechin, lupeol, lupeol acetate and α-amyrin acetate showed minimum inhibition concentration (MIC) values in the range of 225-900 µg/mL against the bacterial strains. Lupeol, lupeol acetate and α-amyrin acetate showed significant antityrosinase activity against mushroom tyrosinase enzyme with percent inhibition of 67.7%, 66.2% and 62.2%, respectively.Keywords: antibacterial, antityrosinase, chemical constituents, Ficus platyphylla
Procedia PDF Downloads 2691976 Exploring Fluoroquinolone-Resistance Dynamics Using a Distinct in Vitro Fermentation Chicken Caeca Model
Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.
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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model
Procedia PDF Downloads 641975 Effects of the Coagulation Bath and Reduction Process on SO2 Adsorption Capacity of Graphene Oxide Fiber
Authors: Özge Alptoğa, Nuray Uçar, Nilgün Karatepe Yavuz, Ayşen Önen
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Sulfur dioxide (SO2) is a very toxic air pollutant gas and it causes the greenhouse effect, photochemical smog, and acid rain, which threaten human health severely. Thus, the capture of SO2 gas is very important for the environment. Graphene which is two-dimensional material has excellent mechanical, chemical, thermal properties, and many application areas such as energy storage devices, gas adsorption, sensing devices, and optical electronics. Further, graphene oxide (GO) is examined as a good adsorbent because of its important features such as functional groups (epoxy, carboxyl and hydroxyl) on the surface and layered structure. The SO2 adsorption properties of the fibers are usually investigated on carbon fibers. In this study, potential adsorption capacity of GO fibers was researched. GO dispersion was first obtained with Hummers’ method from graphite, and then GO fibers were obtained via wet spinning process. These fibers were converted into a disc shape, dried, and then subjected to SO2 gas adsorption test. The SO2 gas adsorption capacity of GO fiber discs was investigated in the fields of utilization of different coagulation baths and reduction by hydrazine hydrate. As coagulation baths, single and triple baths were used. In single bath, only ethanol and CaCl2 (calcium chloride) salt were added. In triple bath, each bath has a different concentration of water/ethanol and CaCl2 salt, and the disc obtained from triple bath has been called as reference disk. The fibers which were produced with single bath were flexible and rough, and the analyses show that they had higher SO2 adsorption capacity than triple bath fibers (reference disk). However, the reduction process did not increase the adsorption capacity, because the SEM images showed that the layers and uniform structure in the fiber form were damaged, and reduction decreased the functional groups which SO2 will be attached. Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD) analyzes were performed on the fibers and discs, and the effects on the results were interpreted. In the future applications of the study, it is aimed that subjects such as pH and additives will be examined.Keywords: coagulation bath, graphene oxide fiber, reduction, SO2 gas adsorption
Procedia PDF Downloads 3601974 Inflammatory Markers in the Blood and Chronic Periodontitis
Authors: Saimir Heta, Ilma Robo, Nevila Alliu, Tea Meta
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Background: Plasma levels of inflammatory markers are the expression of the infectious wastes of existing periodontitis, as well as of existing inflammation everywhere in the body. Materials and Methods: The study consists of the clinical part of the measurement of inflammatory markers of 23 patients diagnosed with chronic periodontitis and the recording of parental periodontal parameters of patient periodontal status: hemorrhage index and probe values, before and 7-10 days after non-surgical periodontal treatment. Results: The level of fibrinogen drops according to the categorization of disease progression, active and passive, with the biggest % (18%-30%) at the fluctuation 10-20 mg/d. Fluctuations in fibrinogen level according to the age of patients in the range 0-10 mg/dL under 40 years and over 40 years was 13%-26%, in the range 10-20 mg/dL was 26%-22%, in the 20-40 mg/dL was 9%-4%. Conclusions: Non-surgical periodontal treatment significantly reduces the level of non-inflammatory markers in the blood. Oral health significantly reduces the potential source for periodontal bacteria, with the potential of promoting thromboembolism, through interaction between thrombocytes.Keywords: chronic periodontitis, atherosclerosis, risk factor, inflammatory markers
Procedia PDF Downloads 1261973 Prevalence and Molecular Characterization of Extended-Spectrum–β Lactamase and Carbapenemase-Producing Enterobacterales from Tunisian Seafood
Authors: Mehdi Soula, Yosra Mani, Estelle Saras, Antoine Drapeau, Raoudha Grami, Mahjoub Aouni, Jean-Yves Madec, Marisa Haenni, Wejdene Mansour
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Multi-resistance to antibiotics in gram-negative bacilli and particularly in enterobacteriaceae, has become frequent in hospitals in Tunisia. However, data on antibiotic resistant bacteria in aquatic products are scarce. The aims of this study are to estimate the proportion of ESBL- and carbapenemase-producing Enterobacterales in seafood (clams and fish) in Tunisia and to molecularly characterize the collected isolates. Two types of seafood were sampled in unrelated markets in four different regions in Tunisia (641 pieces of farmed fish and 1075 mediterranean clams divided into 215 pools, and each pool contained 5 pieces). Once purchased, all samples were incubated in tubes containing peptone salt broth for 24 to 48h at 37°C. After incubation, overnight cultures were isolated on selective MacConkey agar plates supplemented with either imipenem or cefotaxime, identified using API20E test strips (bioMérieux, Marcy-l’Étoile, France) and confirmed by Maldi-TOF MS. Antimicrobial susceptibility was determined by the disk diffusion method on Mueller-Hinton agar plates and results were interpreted according to CA-SFM 2021. ESBL-producing Enterobacterales were detected using the Double Disc Synergy Test (DDST). Carbapenem-resistance was detected using an ertapenem disk and was respectively confirmed using the ROSCO KPC/MBL and OXA-48 Confirm Kit (ROSCO Diagnostica, Taastrup, Denmark). DNA was extracted using a NucleoSpin Microbial DNA extraction kit (Macherey-Nagel, Hoerdt, France), according to the manufacturer’s instructions. Resistance genes were determined using the CGE online tools. The replicon content and plasmid formula were identified from the WGS data using PlasmidFinder 2.0.1 and pMLST 2.0. From farmed fishes, nine ESBL-producing strains (9/641, 1.4%) were isolated, which were identified as E. coli (n=6) and K. pneumoniae (n=3). Among the 215 pools of 5 clams analyzed, 18 ESBL-producing isolates were identified, including 14 E. coli and 4 K. pneumoniae. A low isolation rate of ESBL-producing Enterobacterales was detected 1.6% (18/1075) in clam pools. In fish, the ESBL phenotype was due to the presence of the blaCTX-M-15 gene in all nine isolates, but no carbapenemase gene was identified. In clams, the predominant ESBL phenotype was blaCTX-M-1 (n=6/18). blaCPE (NDM1, OXA48) was detected only in 3 isolates ‘K. pneumoniae isolates’. Replicon typing on the strains carring the ESBL and carbapenemase gene revelead that the major type plasmid carried ESBL were IncF (42.3%) [n=11/26]. In all, our results suggest that seafood can be a reservoir of multi-drug resistant bacteria, most probably of human origin but also by the selection pressure of antibiotic. Our findings raise concerns that seafood bought for consumption may serve as potential reservoirs of AMR genes and pose serious threat to public health.Keywords: BLSE, carbapenemase, enterobacterales, tunisian seafood
Procedia PDF Downloads 1111972 Efficiency Enhancement of Blue OLED by Incorporating Ag Nanoplate Layers
Authors: So-Jeong Kim, Nak-Kwan Chung, Jintae Kim, Juyoung Yun
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The metal nanoplates are potentially used for electroluminescence enhancement of OLEDs owing to the localized surface plasmon resonance. In our study, enhanced electroluminescence in blue organic light-emitting diodes is demonstrated by incorporating silver nanoplates into poly(3,4-ethylene dioxythiophene):polystyrene sulfonic acid. To have surface plasmon resonance absorption peak matching with photoluminescent (PL) peak of blue, Ag nanoplates with triangular shape are used in this study. Finally, about 30 % enhancement in electroluminescence intensity and current efficiency for blue emission devices is obtained via Ag nanoplates.Keywords: efficiency enhancement, nanoplate, OLED, surface plasmon resonance
Procedia PDF Downloads 3441971 Spatial Organization of Cells over the Process of Pellicle Formation by Pseudomonas alkylphenolica KL28
Authors: Kyoung Lee
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Numerous aerobic bacteria have the ability to form multicellular communities on the surface layer of the air-liquid (A-L) interface as a biofilm called a pellicle. Pellicles occupied at the A-L interface will benefit from the utilization of oxygen from air and nutrient from liquid. Buoyancy of cells can be obtained by high surface tension at the A-L interface. Thus, formation of pellicles is an adaptive advantage in utilization of excess nutrients in the standing culture where oxygen depletion is easily set up due to rapid cell growth. In natural environments, pellicles are commonly observed on the surface of lake or pond contaminated with pollutants. Previously, we have shown that when cultured in standing LB media an alkylphenol-degrading bacteria Pseudomonas alkylphenolia KL28 forms pellicles in a diameter of 0.3-0.5 mm with a thickness of ca 40 µm. The pellicles have unique features for possessing flatness and unusual rigidity. In this study, the biogenesis of the circular pellicles has been investigated by observing the cell organization at early stages of pellicle formation and cell arrangements in pellicle, providing a clue for highly organized cellular arrangement to be adapted to the air-liquid niche. Here, we first monitored developmental patterns of pellicle from monolayer to multicellular organization. Pellicles were shaped by controlled growth of constituent cells which accumulate extracellular polymeric substance. The initial two-dimensional growth was transited to multilayers by a constraint force of accumulated self-produced extracellular polymeric substance. Experiments showed that pellicles are formed by clonal growth and even with knock-out of genes for flagella and pilus formation. In contrast, the mutants in the epm gene cluster for alginate-like polymer biosynthesis were incompetent in cell alignment for initial two-dimensional growth of pellicles. Electron microscopic and confocal laser scanning microscopic studies showed that the fully matured structures are highly packed by matrix-encased cells which have special arrangements. The cells on the surface of the pellicle lie relatively flat and inside longitudinally cross packed. HPLC analysis of the extrapolysaccharide (EPS) hydrolysate from the colonies from LB agar showed a composition with L-fucose, L-rhamnose, D-galactosamine, D-glucosamine, D-galactose, D-glucose, D-mannose. However, that from pellicles showed similar neutral and amino sugar profile but missing galactose. Furthermore, uronic acid analysis of EPS hydrolysates by HPLC showed that mannuronic acid was detected from pellicles not from colonies, indicating the epm-derived polymer is critical for pellicle formation as proved by the epm mutants. This study verified that for the circular pellicle architecture P. alkylphenolica KL28 cells utilized EPS building blocks different from that used for colony construction. These results indicate that P. alkylphenolica KL28 is a clever architect that dictates unique cell arrangements with selected EPS matrix material to construct sophisticated building, circular biofilm pellicles.Keywords: biofilm, matrix, pellicle, pseudomonas
Procedia PDF Downloads 1551970 Investigation of Linezolid, 127I-Linezolid and 131I-Linezolid Effects on Slime Layer of Staphylococcus with Nuclear Methods
Authors: Hasan Demiroğlu, Uğur Avcıbaşı, Serhan Sakarya, Perihan Ünak
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Implanted devices are progressively practiced in innovative medicine to relieve pain or improve a compromised function. Implant-associated infections represent an emerging complication, caused by organisms which adhere to the implant surface and grow embedded in a protective extracellular polymeric matrix, known as a biofilm. In addition, the microorganisms within biofilms enter a stationary growth phase and become phenotypically resistant to most antimicrobials, frequently causing treatment failure. In such cases, surgical removal of the implant is often required, causing high morbidity and substantial healthcare costs. Staphylococcus aureus is the most common pathogen causing implant-associated infections. Successful treatment of these infections includes early surgical intervention and antimicrobial treatment with bactericidal drugs that also act on the surface-adhering microorganisms. Linezolid is a promising anti-microbial with ant-staphylococcal activity, used for the treatment of MRSA infections. Linezolid is a synthetic antimicrobial and member of oxazolidinoni group, with a bacteriostatic or bactericidal dose-dependent antimicrobial mechanism against gram-positive bacteria. Intensive use of antibiotics, have emerged multi-resistant organisms over the years and major problems have begun to be experienced in the treatment of infections occurred with them. While new drugs have been developed worldwide, on the other hand infections formed with microorganisms which gained resistance against these drugs were reported and the scale of the problem increases gradually. Scientific studies about the production of bacterial biofilm increased in recent years. For this purpose, we investigated the activity of Lin, Lin radiolabeled with 131I (131I-Lin) and cold iodinated Lin (127I-Lin) against clinical strains of Staphylococcus aureus DSM 4910 in biofilm. In the first stage, radio and cold labeling studies were performed. Quality-control studies of Lin and iodo (radio and cold) Lin derivatives were carried out by using TLC (Thin Layer Radiochromatography) and HPLC (High Pressure Liquid Chromatography). In this context, it was found that the binding yield was obtained to be about 86±2 % for 131I-Lin. The minimal inhibitory concentration (MIC) of Lin, 127I-Lin and 131I-Lin for Staphylococcus aureus DSM 4910 strain were found to be 1µg/mL. In time-kill studies of Lin, 127I-Lin and 131I-Lin were producing ≥ 3 log10 decreases in viable counts (cfu/ml) within 6 h at 2 and 4 fold of MIC respectively. No viable bacteria were observed within the 24 h of the experiments. Biofilm eradication of S. aureus started with 64 µg/mL of Lin, 127I-Lin and 131I-Lin, and OD630 was 0.507±0.0.092, 0.589±0.058 and 0.266±0.047, respectively. The media control of biofilm producing Staphylococcus was 1.675±0,01 (OD630). 131I and 127I did not have any effects on biofilms. Lin and 127I-Lin were found less effectively than 131I-Lin at killing cells in biofilm and biofilm eradication. Our results demonstrate that the 131I-Lin have potent anti-biofilm activity against S. aureus compare to Lin, 127I-Lin and media control. This is suggested that, 131I may have harmful effect on biofilm structure.Keywords: iodine-131, linezolid, radiolabeling, slime layer, Staphylococcus
Procedia PDF Downloads 5581969 A Critical Look on Clustered Regularly Interspaced Short Palindromic Repeats Method Based on Different Mechanisms
Authors: R. Sulakshana, R. Lakshmi
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Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR associate (CRISPR/Cas) is an adaptive immunity system found in bacteria and archaea. It has been modified to serve as a potent gene editing tool. Moreover, it has found widespread use in the field of genome research because of its accessibility and low cost. Several bioinformatics methods have been created to aid in the construction of specific single guide RNA (sgRNA), which is highly active and crucial to CRISPR/Cas performance. Various Cas proteins, including Cas1, Cas2, Cas9, and Cas12, have been used to create genome engineering tools because of their programmable sequence specificity. Class 1 and 2 CRISPR/Cas systems, as well as the processes of all known Cas proteins (including Cas9 and Cas12), are discussed in this review paper. In addition, the various CRISPR methodologies and their tools so far discovered are discussed. Finally, the challenges and issues in the CRISPR system along with future works, are presented.Keywords: gene editing tool, Cas proteins, CRISPR, guideRNA, programmable sequence
Procedia PDF Downloads 1061968 Anticandidal and Antibacterial Silver and Silver(Core)-Gold(Shell) Bimetallic Nanoparticles by Fusarium graminearum
Authors: Dipali Nagaonkar, Mahendra Rai
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Nanotechnology has experienced significant developments in engineered nanomaterials in the core-shell arrangement. Nanomaterials having nanolayers of silver and gold are of primary interest due to their wide applications in catalytical and biomedical fields. Further, mycosynthesis of nanoparticles has been proved as a sustainable synthetic approach of nanobiotechnology. In this context, we have synthesized silver and silver (core)-gold (shell) bimetallic nanoparticles using a fungal extract of Fusarium graminearum by sequential reduction. The core-shell deposition of nanoparticles was confirmed by the red shift in the surface plasmon resonance from 434 nm to 530 nm with the aid of the UV-Visible spectrophotometer. The mean particle size of Ag and Ag-Au nanoparticles was confirmed by nanoparticle tracking analysis as 37 nm and 50 nm respectively. Quite polydispersed and spherical nanoparticles are evident by TEM analysis. These mycosynthesized bimetallic nanoparticles were tested against some pathogenic bacteria and Candida sp. The antimicrobial analysis confirmed enhanced anticandidal and antibacterial potential of bimetallic nanoparticles over their monometallic counterparts.Keywords: bimetallic nanoparticles, core-shell arrangement, mycosynthesis, sequential reduction
Procedia PDF Downloads 5751967 Nitrogen Fixation, Cytokinin and Exopolysachharide Production by indigenous Azotobacter spp. from East Nusa Tenggara
Authors: Reginawanti Hindersah, Widiya Septiani Perdanawati, Dewi Azizah Sulaksana, Hidiyah Ayu Ma’rufah
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Maize in some region in East Nusa Tenggara Indonesia bordering Republic Democratic of Timor Leste is important local food crop and commonly cultivated using conventional method without appropriate plant nutrition system so that productivity is still low. A way to enhance local corn yield is adding biofertilizer containing nitrogen (N2) fixing bacteria such as Azotobacter. The purpose of this research was to determine N2 fixation, cytokinin as well as exopolysachharide production capacity of six indigenous Azotobacter strains in pure culture. The N2 fixation capacities of native 3 day old Azotobacter strains added to Ashby Media varied from 0.01 to 0.39 µM/g/hour. Cytokinin production of these strain in liquid culture of N-free Media was 0.11 to 40.04 ppm while exopolysachharide content in liquid culture of Vermani Media varied from 0.4 to 27.3 g/L. This results demonstrate that some local Azotobacter strains might be used as biofertilizer.Keywords: azotobacter, local isolate, N fixation, phythohormone, exopolysaccaride
Procedia PDF Downloads 4351966 Yeasts Associated to Spontaneous Date Vinegar Process
Authors: F. Halladj, H. Amellal, S. Benamara
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Current consumer trends go towards natural products defined as the products obtained by a traditional manufacturing method. Vinegar is one of those products marketed; it may be industrially obtained by a submerged (fast) or traditional (slow) processes. The latter exhibited a high quality because of its complex microbiological transformations (or two-stage fermentation) by the native must flora. Moreover, although that Acetic acid bacteria have traditionally been considered to play the leading role in vinegar production, some studies have recently highlighted that also yeasts metabolism can affect traditional vinegar chemical properties in a remarkable way. Thus, the aim of this study was to monitor a traditional slow process of vinegar as applied in the south of Algeria using date with hard texture (Degla-Beida variety) to isolate and identify the involved yeasts in order to select them as starter culture. Phenotypic and molecular analysis show that the non-Saccharomyces were the main yeasts species isolated throughout the alcoholic spontaneous fermentation and they included Hanseniaspora guilliermondii and Torulaspora delbrueckii.Keywords: date vinegar, traditional production, yeasts, Phenotypic, Algeria
Procedia PDF Downloads 4311965 Study of Secondary Metabolites of Sargassum Algae: Anticorrosive and Antibacterial Activities
Authors: Prescilla Lambert, Christophe Roos, Mounim Lebrini
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For several years, the Caribbean islands and West Africa have had to deal with the massive arrival of the brown seaweed Sargassum. Overall, this macroalgae, which constitutes a habitat for a great diversity of marine organisms, is also an additional stress factor for the marine environment (e.g., coral reefs). In addition, the accumulation followed by the significant decomposition of the Sargassum spp. biomass on the coast leads to the release of toxic gases (H₂S and NH₃), which calls into question the functioning of the economic, health and tourist life of the island and the other interested territories. Originally, these algae are formed by the eutrophication of the oceans accentuated by global warming. Unfortunately, scientists predict a significant recurrence of these Sargassum strandings for years to come. It is therefore more than necessary to find solutions by putting in place a sustainable management plan for this phenomenon. Martinique, a small island in the Caribbean arc, is one of the many areas impacted by Sargassum seaweed strandings. Since 2011, there has been a constant increase in the degradation of the materials present in this region, largely due to toxic/corrosive gases released by the algae decomposition. In order to protect the structures and the vulnerable building materials while limiting the use of synthetic/petroleum based molecules as much as possible, research is being conducted on molecules of natural origin. Thus, thanks to the chemical composition, which comprise molecules with interesting properties, algae such as Sargassum could potentially help to solve many issues. Therefore, this study focuses on the green extraction and characterization of molecules from the species Sargassum fluitans and Sargassum natans present in Martinique. The secondary metabolites found in these extracts showed variability in yield rates due to local climatic conditions. The tests carried out shed light on the anticorrosive and antibacterial potential of the algae. These extracts can thus be described as natural inhibitors. The effect of variation in inhibitor concentrations was tested in electrochemistry using electrochemical impedance spectroscopy and polarization curves. The analysis of electrochemical results obtained by direct immersion in the extracts and self-assembled molecular layers (SAMs) for Sargassum fluitans III, Sargassum natans I and VIII species was conclusive in acid and alkaline environments. The excellent results obtained reveal an inhibitory efficacy of 88% at 50mg/L for the crude extract of Sargassum fluitans III and efficacies greater than 97% for the chemical families of Sargassum fluitans III. Similarly, microbiological tests also suggest a bactericidal character. Results for Sargassum fluitans III crude extract show a minimum inhibitory concentration (MIC) of 0.005 mg/mL on Gram-negative bacteria and a MIC greater than 0.6 mg/mL on Gram-positive bacteria. These results make it possible to consider the management of local and international issues while valuing a biomass rich in biodegradable molecules. The next step in this study will therefore be the evaluation of the toxicity of Sargassum spp..Keywords: Sargassum, secondary metabolites, anticorrosive, antibacterial, natural inhibitors
Procedia PDF Downloads 731964 Biophysical and Structural Characterization of Transcription Factor Rv0047c of Mycobacterium Tuberculosis H37Rv
Authors: Md. Samsuddin Ansari, Ashish Arora
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Every year 10 million people fall ill with one of the oldest diseases known as tuberculosis, caused by Mycobacterium tuberculosis. The success of M. tuberculosis as a pathogen is because of its ability to persist in host tissues. Multidrug resistance (MDR) mycobacteria cases increase every day, which is associated with efflux pumps controlled at the level of transcription. The transcription regulators of MDR transporters in bacteria belong to one of the following four regulatory protein families: AraC, MarR, MerR, and TetR. Phenolic acid decarboxylase repressor (PadR), like a family of transcription regulators, is closely related to the MarR family. Phenolic acid decarboxylase repressor (PadR) was first identified as a transcription factor involved in the regulation of phenolic acid stress response in various microorganisms (including Mycobacterium tuberculosis H37Rv). Recently research has shown that the PadR family transcription factors are global, multifunction transcription regulators. Rv0047c is a PadR subfamily-1 protein. We are exploring the biophysical and structural characterization of Rv0047c. The Rv0047 gene was amplified by PCR using the primers containing EcoRI and HindIII restriction enzyme sites cloned in pET-NH6 vector and overexpressed in DH5α and BL21 (λDE3) cells of E. coli following purification with Ni2+-NTA column and size exclusion chromatography. We did DSC to know the thermal stability; the Tm (transition temperature) of protein is 55.29ºC, and ΔH (enthalpy change) of 6.92 kcal/mol. Circular dichroism to know the secondary structure and conformation and fluorescence spectroscopy for tertiary structure study of protein. To understand the effect of pH on the structure, function, and stability of Rv0047c we employed spectroscopy techniques such as circular dichroism, fluorescence, and absorbance measurements in a wide range of pH (from pH-2.0 to pH-12). At low and high pH, it shows drastic changes in the secondary and tertiary structure of the protein. EMSA studies showed the specific binding of Rv0047c with its own 30-bp promoter region. To determine the effect of complex formation on the secondary structure of Rv0047c, we examined the CD spectra of the complex of Rv0047c with promoter DNA of rv0047. The functional role of Rv0047c was characterized by over-expressing the Rv0047c gene under the control of hsp60 promoter in Mycobacterium tuberculosis H37Rv. We have predicted the three-dimensional structure of Rv0047c using the Swiss Model and Modeller, with validity checked by the Ramachandra plot. We did molecular docking of Rv0047c with dnaA, through PatchDock following refinement through FireDock. Through this, it is possible to easily identify the binding hot-stop of the receptor molecule with that of the ligand, the nature of the interface itself, and the conformational change undergone by the protein pattern. We are using X-crystallography to unravel the structure of Rv0047c. Overall the studies show that Rv0047c may have transcription regulation along with providing an insight into the activity of Rv0047c in the pH range of subcellular environment and helps to understand the protein-protein interaction, a novel target to kill dormant bacteria and potential strategy for tuberculosis control.Keywords: mycobacterium tuberculosis, phenolic acid decarboxylase repressor, Rv0047c, Circular dichroism, fluorescence spectroscopy, docking, protein-protein interaction
Procedia PDF Downloads 1211963 Coupling of Microfluidic Droplet Systems with ESI-MS Detection for Reaction Optimization
Authors: Julia R. Beulig, Stefan Ohla, Detlev Belder
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In contrast to off-line analytical methods, lab-on-a-chip technology delivers direct information about the observed reaction. Therefore, microfluidic devices make an important scientific contribution, e.g. in the field of synthetic chemistry. Herein, the rapid generation of analytical data can be applied for the optimization of chemical reactions. These microfluidic devices enable a fast change of reaction conditions as well as a resource saving method of operation. In the presented work, we focus on the investigation of multiphase regimes, more specifically on a biphasic microfluidic droplet systems. Here, every single droplet is a reaction container with customized conditions. The biggest challenge is the rapid qualitative and quantitative readout of information as most detection techniques for droplet systems are non-specific, time-consuming or too slow. An exception is the electrospray mass spectrometry (ESI-MS). The combination of a reaction screening platform with a rapid and specific detection method is an important step in droplet-based microfluidics. In this work, we present a novel approach for synthesis optimization on the nanoliter scale with direct ESI-MS detection. The development of a droplet-based microfluidic device, which enables the modification of different parameters while simultaneously monitoring the effect on the reaction within a single run, is shown. By common soft- and photolithographic techniques a polydimethylsiloxane (PDMS) microfluidic chip with different functionalities is developed. As an interface for the MS detection, we use a steel capillary for ESI and improve the spray stability with a Teflon siphon tubing, which is inserted underneath the steel capillary. By optimizing the flow rates, it is possible to screen parameters of various reactions, this is exemplarity shown by a Domino Knoevenagel Hetero-Diels-Alder reaction. Different starting materials, catalyst concentrations and solvent compositions are investigated. Due to the high repetition rate of the droplet production, each set of reaction condition is examined hundreds of times. As a result, of the investigation, we receive possible reagents, the ideal water-methanol ratio of the solvent and the most effective catalyst concentration. The developed system can help to determine important information about the optimal parameters of a reaction within a short time. With this novel tool, we make an important step on the field of combining droplet-based microfluidics with organic reaction screening.Keywords: droplet, mass spectrometry, microfluidics, organic reaction, screening
Procedia PDF Downloads 3021962 Nanocomposite Metal Material: Study of Antimicrobial and Catalytic Properties
Authors: Roman J. Jedrzejczyk, Damian K. Chlebda, Anna Dziedzicka, Rafal Wazny, Agnieszka Domka, Maciej Sitarz, Przemyslaw J. Jodlowski
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The aim of this study was to obtain antimicrobial material based on thin zirconium dioxide coatings on structured reactors doped with metal nanoparticles using the sonochemical sol-gel method. As a result, dense, uniform zirconium dioxide films were obtained on the kanthal sheets which can be used as support materials in antimicrobial converters with sophisticated shapes. The material was characterised by physicochemical methods, such as AFM, SEM, EDX, XRF, XRD, XPS and in situ Raman and DRIFT spectroscopy. In terms of antimicrobial activity, the material was tested by ATP/AMP method using model microbes isolated from the real systems. The results show that the material can be potentially used in the market as a good candidate for active package and as active bulkheads of climatic systems. The mechanical tests showed that the developed method is an efficient way to obtain durable converters with high antimicrobial activity against fungi and bacteria.Keywords: antimicrobial properties, kanthal steel, nanocomposite, zirconium oxide
Procedia PDF Downloads 2021961 Fuelwood Heating, Felling, Energy Renewing in Total Fueling of Fuelwood, Renewable Technologies
Authors: Adeiza Matthew, Oluwamishola Abubakar
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In conclusion, Fuelwood is a traditional and renewable source of energy that can have both positive and negative impacts. Adopting sustainable practices for its collection, transportation, and use and investing in renewable technologies can help mitigate the negative effects and provide a clean and reliable source of energy, improve living standards and support economic development. For example, solar energy can be used to generate electricity, heat homes and water, and can even be used for cooking. Wind energy can be used to generate electricity, and geothermal energy can be used for heating and cooling. Biogas can be produced from waste products such as animal manure, sewage, and organic kitchen waste and can be used for cooking and lighting.Keywords: calorific, BTU, wood moisture content, density of wood
Procedia PDF Downloads 1091960 Outcomes of Combined Penetrating keratoplasty and Vitreo-Retinal Surgery in Management of Endophthalmitis with Obscured Corneal Clarity
Authors: Abhishek Dave, Manisha Singh
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Purpose: The study aims to evaluate the outcomes of combined Penetrating keratoplasty (PKP) and Vitreo-Retinal (VR) surgery in patients having endophthalmitis with poor corneal clarity. Methods: PKP with VR Surgery was performed in 43 eyes. This is a retrospective analysis of their preoperative, intraoperative and microbiological characteristics and anatomical and functional outcomes. Results: Corneal opacification was due to corneal ulcer in 30 (69.7%), graft infection in 8 (18.6%), bullous keratopathy in 4 and corneal scar in 1 eye. Postoperative visual acuity improved in 20 (46.5%), not changed in 14 (32.5%) and deteriorated in 9 eyes (20.9%). Poor anatomic success was seen in 15 (34.88%) eyes (9-phthisis bulbi, 6-eviscerated). Graft remained clear in 24 eyes (1 year). Microbiology revealed bacteria in 26, fungus in 14 and no growth in 3 eyes. Six out of 11 patients having poor vision in the fellow eye, too, achieved functional success. Conclusion: PKP with VR surgery is a complex but globe-salvaging procedure for poor prognosis eyes, which otherwise would need evisceration.Keywords: penetrating keratoplasty, VR surgery, endophthalmitis, corneal ulcer
Procedia PDF Downloads 521959 Detection and Quantification of Viable but Not Culturable Vibrio Parahaemolyticus in Frozen Bivalve Molluscs
Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino
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Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus
Procedia PDF Downloads 1401958 Spectrophotometric Determination of 5-Aminosalicylic Acid in Pharmaceutical Samples
Authors: Chand Pasha
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A Simple, accurate and precise spectrophotometric method for the quantitative analysis of determination of 5-aminosalicylic acid is described. This method is based on the reaction of 5-aminosalicylic acid with nitrite in acid medium to form diazonium ion, which is coupled with acetylacetone in basic medium to form azo dyes, which shows absorption maxima at 470 nm. The method obeys Beer’s law in the concentration range of 0.5-11.2 gml-1 of 5-aminosalicylic acid with acetylacetone. The molar absorptivity and Sandell’s sensitivity of 5-aminosalicylic acid -acetylacetone azo dye is 2.672 ×104 lmol-1cm-1, 5.731 × 10-3 gcm-2 respectively. The dye formed is stable for 10 hrs. The optimum reaction conditions and other analytical parameters are evaluated. Interference due to foreign organic compounds have been investigated. The method has been successfully applied to the determination of 5-aminosalicylic acid in pharmaceutical samples.Keywords: spectrophotometry, diazotization, mesalazine, nitrite, acetylacetone
Procedia PDF Downloads 1911957 Fast Fashion Parallel to Sustainable Fashion in India
Authors: Saurav Sharma, Deepshikha Sharma, Pratibha Sharma
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This paper includes fast fashion verses sustainable fashion or slow fashion Indian based consumers. The expression ‘Fast fashion’ is generally referred to low-cost clothing collections that considered first hand copy of luxury brands, sometime interchangeably used with ‘mass fashion’. Whereas slow fashion or limited fashion which are consider to be more organic or eco-friendly. "Sustainable fashion is ethical fashion and here the consumer is just not design conscious but also social-environment conscious". Paper will deal with desire of young Indian consumer towards such luxury brands present in India, and their understanding of sustainable fashion, how to maintain the equilibrium between never newer fashion, style, and fashion sustainability.Keywords: fast fashion, sustainable fashion, sustainability, India
Procedia PDF Downloads 775