Search results for: coupled enzyme system of bioluminescence bacteria NAD(P)H:FMN-oxidoreductase–luciferase

Authors: Young-Hun Lee, Tae-Gwan Kim, Jong-Kyu Kim, Young-Chul Park

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In this study, Spring safety valve Heat - structure coupled analysis was carried out. Full analysis procedure and performing thermal analysis at a maximum temperature, them to the results obtained through to give an additional load and the pressure on the valve interior, and Disc holder Heat-Coupled structure Analysis was carried out. Modeled using a 3D design program Solidworks, For the modeling of the safety valve was used 3D finite element analysis program ANSYS. The final result to be obtained through the Analysis examined the stability of the maximum displacement and the maximum stress to the valve internal components occurring in the high-pressure conditions.

Keywords: finite element method, spring safety valve, gap, stress, strain, deformation

Procedia PDF Downloads 359

Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

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Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

Procedia PDF Downloads 85

Authors: Asowata Osamede, Christo Pienaar, Johan Bekker

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Mismatch in electrical energy (power) or outage from commercial providers, in general, does not promote development to the public and private sector, these basically limit the development of industries. The necessity for a well-structured photovoltaic (PV) system is of importance for an efficient and cost-effective monitoring system. The major renewable energy potential on earth is provided from solar radiation and solar photovoltaics (PV) are considered a promising technological solution to support the global transformation to a low-carbon economy and reduction on the dependence on fossil fuels. Solar arrays which consist of various PV module should be operated at the maximum power point in order to reduce the overall cost of the system. So power regulation and conditioning circuits should be incorporated in the set-up of a PV system. Power regulation circuits used in PV systems include maximum power point trackers, DC-DC converters and solar chargers. Inappropriate choice of power conditioning device in a basic off-grid PV system can attribute to power loss, hence the need for a right choice of power conditioning device to be coupled with the system of the essence. This paper presents the design and implementation of a power conditioning devices in order to improve the overall yield from the availability of solar energy and the system’s total efficiency. The power conditioning devices taken into consideration in the project includes the Buck and Boost DC-DC converters as well as solar chargers with MPPT. A logging interface circuit (LIC) is designed and employed into the system. The LIC is designed on a printed circuit board. It basically has DC current signalling sensors, specifically the LTS 6-NP. The LIC is consequently required to program the voltages in the system (these include the PV voltage and the power conditioning device voltage). The voltage is structured in such a way that it can be accommodated by the data logger. Preliminary results which include availability of power as well as power loss in the system and efficiency will be presented and this would be used to draw the final conclusion.

Keywords: tilt and orientation angles, solar chargers, PV panels, storage devices, direct solar radiation

Procedia PDF Downloads 130

Authors: Qijiao He

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MicroElectrical-Mechanical System (MEMS) is one of the most rapidly developing frontier research field both in theory study and applied technology. Micro-channel is a very important link component of MEMS. With the research and development of MEMS, the size of the micro-devices and the micro-channels becomes further smaller. Compared with the macroscale flow, the flow characteristics of gas in the micro-channel have changed, and the rarefaction effect appears obviously. However, for the rarefied gas and microscale flow, Navier-Stokes-Fourier (NSF) equations are no longer appropriate due to the breakup of the continuum hypothesis. A Nonlinear Coupled Constitutive Model (NCCM) has been derived from the Boltzmann equation to describe the characteristics of both continuum and rarefied gas flows. We apply the present scheme to simulate continuum and rarefied gas flows in a micro-channel structure. And for comparison, we apply other widely used methods which based on particle simulation or direct solution of distribution function, such as Direct simulation of Monte Carlo (DSMC), Unified Gas-Kinetic Scheme (UGKS) and Lattice Boltzmann Method (LBM), to simulate the flows. The results show that the present solution is in better agreement with the experimental data and the DSMC, UGKS and LBM results than the NSF results in rarefied cases but is in good agreement with the NSF results in continuum cases. And some characteristics of both continuum and rarefied gas flows are observed and analyzed.

Keywords: continuum and rarefied gas flows, discontinuous Galerkin method, generalized hydrodynamic equations, numerical simulation

Procedia PDF Downloads 161

Authors: Chika C. Ogueke, Ijeoma M. Agunwah

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The antimicrobial activities and preservative potentials of crude extracts of Alstonia boonei stem bark and Euphorbia hirta leaves were studied. Soxhlet extraction and cold ethanol extraction methods were used for the extraction of the dried and ground plant samples. Well in agar diffusion method was used for the antimicrobial screening at different concentrations of 25mg/ml, 50mg/ml, 100mg/ml and 200mg/ml on E.coli and B.subtilis. The preservative effects of the extracts at 0.1%, 0.2% and 0.3% singly and in combination were determined in minced meat using E. coli and B. subtilis as test isolates. Phytochemical analysis was also conducted on the extracts using standard analytical methods. E.hirta cold and A.boonei cold extracts gave the highest zone of growth inhibition on E. coli and B.substilis with 20mm zone diameter at 200mg/ml concentration. Phytochemical analysis revealed the presence of alkaloids, flavonoids, tannins, saponins and cardiac glycosides. A.boonei at 0.1, 0.2 and 0.3% produced a log cycle reduction on the growth of E.coli. Mixture of A. boonei and E. hirta extracts (1:1) at 0.1% and 0.2% also produced a log cycle reduction on the growth of E.coli and B. subtilis, however the A. boonei extracts had more significant effect on the isolates. The observed antimicrobial activities are attributed to the phytochemicals identified in the extracts. The results reveal the potentials of plant extracts as natural antimicrobial preservatives in minced meat. Thus the crude extracts can act as inhibitors of bacteria in a food system. Upon further purification better results may be obtained.

Keywords: antimicrobial preservative, crude extracts, minced meat, test isolates

Procedia PDF Downloads 286

Authors: Ahmed Tawfik

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Oxidation of 1,4 dioxane produces metabolites by-products involving glycolaldehyde and acids that have geno- and cytotoxicity impact on microbial degradation. Thereby, the incorporation of algae with bacteria in the treatment system would eliminate and overcome the accumulation of metabolites that are utilized as a carbon source for the build-up of biomass. Therefore, the aim of the present study is to assess the potential of algae/bacteria-based membrane bioreactor (AB-MBR) for biodegradation of 1,4 dioxane-rich wastewater at a high imposed loading rate. Three identical reactors, i.e., AB-MBR1, AB-MBR2, and AB-MBR3, were operated in parallel at 1,4 dioxane loading rates of 641.7, 320.9, and 160.4 mg/L. d., and HRTs of 6.0, 12 and 24 h. respectively. The AB-MBR1 achieved 1,4 dioxane removal rate of 263.7 mg/L.d., where the residual value in the treated effluent amounted to 94.4±22.9 mg/L. Reducing the 1,4 dioxane loading rate (LR) to 320.9 mg/L.d in the AB-MBR2 maximized the removal rate efficiency of 265.9 mg/L.d., with a removal efficiency of 82.8±3.2%. The minimum value of 1,4 dioxane of 17.3±1.8 mg/L in the treated effluent of AB-MBR3 was obtained at an HRT of 24.0 h and loading rate of 160.4 mg/L.d. The mechanism of 1,4 dioxane degradation in AB-MBR was a combination of volatilization (8.03±0.6%), UV oxidation (14.1±0.9%), microbial biodegradation (49.1±3.9%) and absorption/uptake and assimilation by algae (28.8±2.%). Further, the Thioclava, Afipia, and Mycobacterium genera oxidized and produced the required enzymes for hydrolysis and cleavage of the dioxane ring into 2-hydroxy-1,4 dioxane. Moreover, the fungi, i.e., Basidiomycota and Cryptomycota, played a big role in the degradation of the 1,4 dioxane into 2-hydroxy-1,4 dioxane. Xanthobacter and Mesorhizobium were involved in the metabolism process by secreting alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), and glycolate oxidase. Bacteria and fungi produced dehydrogenase (DH) for the transformation of 2-hydroxy-1,4 dioxane into 2-hydroxy-ethoxyacetaldehyde. The latter is converted into Ethylene glycol by Aldehyde hydrogenase (ALDH). Ethylene glycol is oxidized into acids using Alcohol hydrogenase (ADH). The Diatomea, Chlorophyta, and Streptophyta utilize the metabolites for biomass assimilation and produce the required oxygen for further oxidation of the dioxane and its metabolites by-products of bacteria and fungi. The major portion of metabolites (ethylene glycol, glycolic acid, and oxalic acid were removed due to uptake and absorption by algae (43±4.3%), followed by adsorption (18.4±0.9%). The volatilization and UV oxidation contribution for the degradation of metabolites were 8.7±0.7% and 12.3±0.8%, respectively. The capabilities of genera Defluviimonas, Thioclava, Luteolibacter, and Afipia. The genera of Defluviimonas, Thioclava, Luteolibacter, and Mycobacterium were grown under a high 1,4 dioxane LR of 641.7 mg/L.d. The Chlorophyta (4.1-43.6%), Streptophyta (2.5-21.7%), and Diatomea (0.8-1.4%) phyla were dominant for degradation of 1,4 dioxane. The results of this study strongly demonstrated that the bioremediation and bioaugmentation process can safely remove 1,4 dioxane from industrial wastewater while minimizing environmental concerns and reducing economic costs.

Keywords: wastewater, membrane bioreactor, bacterial community, algal community

Procedia PDF Downloads 39

Authors: Vishakha Kaim, Mookan Natarajan, Sandeep Kaur-Ghumaan

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Hydrogen is one of the most abundant elements present on earth’s crust and considered to be the simplest element in existence. It is not found naturally as a gas on earth and thus has to be manufactured. Hydrogen can be produced from a variety of sources, i.e., water, fossil fuels, or biomass and it is a byproduct of many chemical processes. It is also considered as a secondary source of energy commonly referred to as an energy carrier. Though hydrogen is not widely used as a fuel, it still has the potential for greater use in the future as a clean and renewable source of energy. Electrocatalysis is one of the important source for the production of hydrogen which could contribute to this prominent challenge. Metals such as platinum and palladium are considered efficient for hydrogen production but with limited applications. As a result, a wide variety of metal complexes with earth abundant elements and varied ligand environments have been explored for the electrochemical production of hydrogen. In nature, [FeFe] hydrogenase enzyme present in DesulfoVibrio desulfuricans and Clostridium pasteurianum catalyses the reversible interconversion of protons and electrons into dihydrogen. Since the first structure for the enzyme was reported in 1990s, a range of iron complexes has been synthesized as structural and functional mimics of the enzyme active site. Mn is one of the most desirable element for sustainable catalytic transformations, immediately behind Fe and Ti. Only limited number manganese complexes have been reported in the last two decades as catalysts for proton reduction. Furthermore, redox reactions could be carried out in a facile manner, due to the capability of manganese complexes to be stable at different oxidation states. Herein are reported, four µ2-thiolate bridged manganese complexes [Mn₂(CO)₆(μ-S₂N₄C₁₄H₁₀)] 1, [Mn₂(CO)7(μ- S₂N₄C₁₄H₁₀)] 2, Mn₂(CO)₆(μ-S₄N₂C₁₄H₁₀)] 3 and [Mn₂(CO)(μ- S₄N₂C₁₄H₁₀)] 4 have been synthesized and characterized. The cyclic voltammograms of the complexes displayed irreversible reduction peaks in the range - 0.9 to -1.3 V (vs. Fc⁺/Fc in acetonitrile at 0.1 Vs⁻¹). The complexes were catalytically active towards proton reduction in the presence of trifluoroacetic acid as seen from electrochemical investigations.

Keywords: earth abundant, electrocatalytic, hydrogen, manganese

Procedia PDF Downloads 168

Authors: Slimane Mokrani, Nabti El hafid

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Description of the subject: Currently, several efforts focus on the study of biodiversity, microbial biotechnology, and the use of ecological strategies. Objectives: The aim of this present work is to determine the functional diversity of bacteria in rhizospheric and non-rhizospheric soils of different plants. Methods: Bacteria were isolated from soil and identified based on physiological and biochemical characters and genotypic taxonomy performed by 16S rDNA and BOX-PCR. As well as the characterization of various PGPR traits. Then, they are tested for their effects on the stimulation of seed germination and the growth of Phaseolus vulgaris L. As well as their biological control activities with regard to the phytopathogenic bacterial isolate Xapf. Results and Discussion: The biochemical and physiological identification of 75 bacterial isolates made it possible to associate them with the two groups of fluorescent Pseudomonas (74.67%) and non-fluorescent Pseudomonas (25.33%). The identification by 16S rDNA of 27 strains made it possible to attribute the majority of the strains to the genus Pseudomonas (81.48%), Serratia (7.41%) and Bacillus (11.11%). The bacterial strains showed a high capacity to produce IAA, siderophores, HCN and to solubilize phosphate. A significant stimulation of germination and growth was observed by applying the Pseudomonas strains. Furthermore, significant reductions in the severity and intensity of the disease caused caused by Xapf were observed. Conclusion: The bacteria described in this present study endowed with different PGPR activities seem to be very promising for their uses as biological control agents and bio-fertilization.

Keywords: biofertilization, biological control, phaseolus vulgaris L, pseudomonas, Xanthomonas axonopodis pv. phaseoli var. fuscans and common blight

Procedia PDF Downloads 74

Authors: Ravindra B. Patil, P. Krishnamoorthy, Shriram Sethuraman

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This work proposes a novel Gaussian Mixture Model (GMM) based approach for accurate tracking of the arterial wall and subsequent computation of the distension waveform using Radio Frequency (RF) ultrasound signal. The approach was evaluated on ultrasound RF data acquired using a prototype ultrasound system from an artery mimicking flow phantom. The effectiveness of the proposed algorithm is demonstrated by comparing with existing wall tracking algorithms. The experimental results show that the proposed method provides 20% reduction in the error margin compared to the existing approaches in tracking the arterial wall movement. This approach coupled with ultrasound system can be used to estimate the arterial compliance parameters required for screening of cardiovascular related disorders.

Keywords: distension waveform, Gaussian Mixture Model, RF ultrasound, arterial wall movement

Procedia PDF Downloads 496

Authors: Shubhashree Mahalik, Dhanesh Kumar, Jatin Kumar Pradhan

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Odisha is one of the coastal states situated on the eastern part of India with 480 km long coastline. The coastal Odisha is referred to as "Gift of Six Rivers". Balasore, a major coastal district of Odisha is bounded by Bay of Bengal in the East having 26 km long seashore. It is lined with several estuaries rich in biodiversity.Several studies have been carried out on the macro flora and fauna of this area but very few documented information are available regarding microbial biodiversity. In the present study, an attempt has been made to isolate and identify bacteria found along the estuaries of Balasore.Many marine microorganisms are sources of natural products which makes them potential industrial organisms. So the ability of the isolated bacteria to secrete one such industrially significant product, PHB (Polyhydroxybutyrate) has been elucidated. Several rounds of sampling, pure culture, morphological, biochemical and phylogenetic screening led to the identification of two PHB producing strains. Isolate 5 was identified to be Brevibacillus sp. and has maximum similarity to Brevibacillus parabrevis (KX83268). The isolate was named as Brevibacillus sp.KEI-5. Isolate 8 was identified asLysinibacillus sp. having closest similarity withLysinibacillus boroni-tolerance (KP314269) and named as Lysinibacillus sp. KEI-8.Media, temperature, carbon, nitrogen and salinity requirement were optimized for both isolates. Submerged fermentation of both isolates in Terrific Broth media supplemented with optimized carbon and nitrogen source at 37°C led to significant accumulation of PHB as detected by colorimetric method.

Keywords: Bacillus, estuary, marine, Odisha, polyhydroxy butyrate

Procedia PDF Downloads 345

Authors: Emmanuel Ekpa, Adaji Andrew, Queen Opaluwa, Isreal Daraobong

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Three (3) indigenous bacteria species (Bacillus spp, Acinectobacter spp and Moraxella spp) previously isolated from contaminated soil of some auto mechanic workshops were used for bioremediation studies on some irrigation water used at Sarkin-noma Fadama farms located in Lokoja Kogi State, Nigeria. This was done in order to investigate their bioremediation potentials using a simple pour plate method. The physicochemical parameters and heavy metal analysis (using AAS iCE 3000) of the irrigation water were performed before and after inoculation of the isolated organisms. Nitrate and phosphate concentration were found to be 10.56mg/L and 12.63mg/L prior to inoculation while iron and zinc were 0.9569mg/L and 0.2245mg/L respectively. Other physicochemical parameters were also observed to be high prior to inoculation. After the bioremediation test (inoculation with the isolated organisms), a nitrate and phosphate content of 2.53mg/L and 2.61mg/L were recorded respectively, iron and zinc gave 0.1694mg/L and 0.0174mg/L concentrations while other physicochemical parameters measured were also found to be lower in their respective values. The implication of this present study is that a number of carefully isolated indigenous bacteria species are capable of reducing the amount of heavy metal concentrations in water. Also, non-metallic contaminants like nitrate and phosphate are susceptible to bioremediation in the presence of such efficient system.

Keywords: bioremediation, heavy metals, physicochemical parameters, Bacillus spp, Acinectobacter spp and Moraxella spp, AAS, spectrometer 3000

Procedia PDF Downloads 330

Authors: Yaara Oppenheimer-Shaanan, Nimrod Nachmias, Marina Campos Rocha, Neta Schlezinger, Noam Dotan, Asaf Levy

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Fusarium oxysporum, a fungus that attacks a broad range of plants and can cause infections in humans, operates across different kingdoms. This pathogen encounters varied conditions, such as temperature, pH, and nutrient availability, in plant and human hosts. The Fusarium oxysporum species complex, pervasive in soils globally, can affect numerous plants, including key crops like tomatoes and bananas. Controlling Fusarium infections can involve biocontrol agents that hinder the growth of harmful strains. Our research developed a computational method to identify toxin domains within a vast number of microbial genomes, leading to the discovery of nine distinct toxins capable of killing bacteria and fungi, including Fusarium. These toxins appear to function as enzymes, causing significant damage to cellular structures, membranes and DNA. We explored biological control using bacteria that produce polymorphic toxins, finding that certain bacteria, non-pathogenic to plants, offer a safe biological alternative for Fusarium management, as they did not harm macrophage cells or C. elegans. Additionally, we elucidated the 3D structures of two toxins with their protective immunity proteins, revealing their function as unique DNases. These potent toxins are likely instrumental in microbial competition within plant ecosystems and could serve as biocontrol agents to mitigate Fusarium wilt and related diseases.

Keywords: microbial toxins, antifungal, Fusarium oxysporum, bacterial-fungal intreactions

Procedia PDF Downloads 44

Authors: M. Ottenbrite, G. Yilmaz, J. Devenish, M. Kang, H. Dan, M. Lin, C. Lau, C. Carrillo, K. Bessonov, J. Nash, E. Topp, J. Guan

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Consumption of food contaminated by antibiotic-resistant (AR) bacteria may lead to the transmission of AR genes in the gut microbiota and cause AR bacterial infection, a significant public health concern. However, information is limited on if and how background microbes from the food matrix (food microbes) may influence resistance transmission. Thus, we assessed the colonization of a β-lactam resistant Salmonella Heidelberg strain (donor) and a β-lactam susceptible S. Typhimurium strain (recipient) and the transfer of the resistance genes in the mouse gut in the presence or absence of food microbes that were derived from washing freshly-harvested carrots. Mice were pre-treated with streptomycin and then inoculated with both donor and recipient bacteria or recipient only. Fecal shedding of the donor, recipient, and transconjugant bacteria was enumerated using selective culture techniques. Transfer of AR genes was confirmed by whole genome sequencing. Gut microbial composition was determined by 16s rRNA amplicon sequencing. Significantly lower numbers of donors and recipients were shed from mice that were inoculated with food microbes compared to those without food microbe inoculation. S. Typhimurium transconjugants were only recovered from mice without inoculation of food microbes. A significantly higher survival rate was in mice with vs. without inoculation of food microbes. The results suggest that the food microbes may compete with both the donor and recipient Salmonella, limit their growth and reduce transmission of the β-lactam resistance gene in the mouse gut.

Keywords: antibiotic resistance, gene transfer, gut microbiota, Salmonella infection

Procedia PDF Downloads 65

Authors: Amel Bouderhem, Aminata Ould El Hadj Khelil, Amina N. Djrarbaoui, Aroussi Aroussi

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The present work aims to find the influence of the nature of the soil on the effectiveness of the biodegradation of hydrocarbons by a mixture of bacterial strains hydrocarbonoclastes. Processes of bioaugmentation and biostimulation trial are applied to samples of soils polluted voluntarily by the crude oil. For the evaluation of the biodegradation of hydrocarbons, the bacterial load, the pH and organic carbon total are followed in the different experimental batches. He bacterial load of the sandy soil varies among the witnesses of 45,2 .108 CFU/ml at the beginning of the experimentation to 214,07.108 CFU/ml at the end of the experiment. Of the soil silty-clay varies between 103,31 .108 CFU/ml and 614,86.108 CFU/ml . It was found a strong increase in the bacterial biomass during the processing of all samples. This increase is more important in the samples of sand bioaugmente or biomass increased from 63.16 .108 CFU/ml to 309.68 .108 CFU/ml than in soil samples silty clay- bioaugmente whose content in bacteria evolved of 73,01 .108 CFU/ml to 631.80 . 108CFU/ml

Keywords: pollution, hydrocarbons, bioremediation, bacteria hydrocarbonoclastes, ground, texture

Procedia PDF Downloads 465

Authors: P. S. BEDI* , Neavty Thakur, Balvınder Sıngh

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In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3, NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria eg. Bacillus subtilis, Gram negative bacteria eg. E. coli, K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant posseses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Keywords: Girardinia heterophylla, leaves and stem extracts, Antibacterial activity, antifungal activity.

Procedia PDF Downloads 338

Authors: Tzyy-Rong Jinn, Sheng-Kuo Hsieh, Yi-Ching Chung, Feng-Chia Hsieh

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In this study, we attempted to develop a recombinant oleosin-based fusion expression strategy in Escherichia coli (E. coli) and coupled with the artificial oil bodies (AOB)-cyanogen bromide technology platform to produce bioactive mastoparan B (MP-B). As reported, the oleosin in AOB system plays a carrier (fusion with target protein), since oleosin possess two amphipathic regions (at the N-terminus and C-terminus), which result in the N-terminus and C-terminus of oleosin could be arranged on the surface of AOB. Thus, the target protein fused to the N-terminus or C-terminus of oleosin which also is exposed on the surface of AOB, and this process will greatly facilitate the subsequent separation and purification of target protein from AOB. In addition, oleosin, a unique structural protein of seed oil bodies, has the added advantage of helping the fused MP-B expressed in inclusion bodies, which can protect from proteolytic degradation. In this work, MP-B was fused to the C-terminus of oleosin and then was expressed in E. coli as an insoluble recombinant protein. As a consequence, we successfully developed a reliable recombinant oleosin-based fusion expression strategy in Escherichia coli and coupled with the artificial oil bodies (AOB)-cyanogen bromide technology platform to produce the small peptide, MP-B. Take together, this platform provides an insight into the production of active MP-B, which will facilitate studies and applications of this peptide in the future.

Keywords: artificial oil bodies, Escherichia coli, Oleosin-fusion protein, Mastoparan-B

Procedia PDF Downloads 448

Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel Abugri, Boakai Robertson, Olufemi S. Ajayi

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The emergence of multidrug resistance poses a huge risk to public health globally. Yet these recalcitrant pathogens continue to rise in incidence rate, with resistance rates significantly outpacing the speed of antibiotic development. This, therefore, presents an aura of related health issues such as untreatable nosocomial infections arising from organ transplants and surgeries, as well as community-acquired infections that are related to people with compromised immunity, e.g., diabetic and HIV patients, etc. There is a global effort to fight multidrug-resistant pathogens spearheaded by the World Health Organization, thus calling for research into novel antimicrobial agents to fight multiple drug resistance. Previously, our laboratory demonstrated that Cannabidiol (CBD) was an effective antimicrobial against Salmonella typhimurium (S. typhimurium). However, we observed resistance development over time. To understand the mechanisms S. typhimurium uses to develop resistance to Cannabidiol (CBD), we studied the abundance of bacteria lipopolysaccharide (LPS) and membrane sterols of both susceptible and resistant S. typhimurium. Using real-time quantitative polymerase chain reaction (RT-qPCR), we also analyzed the expression of selected genes known for aiding resistance development in S. typhimurium. We discovered that there was a significantly higher expression of blaTEM, fimA, fimZ, and integrons in the CBD-resistant bacteria, and these were also accompanied by a shift in abundance in cell surface molecules such as lipopolysaccharide (LPS) and sterols.

Keywords: antimicrobials, resistance, cannabidiol, gram-negative bacteria, integrons, blaTEM, Fim, LPS, ergosterols

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Authors: Atefeh Esmailnejad, Mohammad Abbaszadeh Hasiri

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Coxiella burnetii is a gram-negative obligatory intracellular bacterium that causes Q fever, a significant zoonotic disease. Sheep, cattle, and goats are the most commonly reported reservoirs for the bacteria, but infected cats and dogs have also been implicated in the transmission of the disease to human. The aim of present study was to investigate the presence of antibodies against Coxiella burnetii among companion dogs in Fars province, South of Iran. A total of 181 blood samples were collected from asymptomatic dogs, mostly referred to Veterinary Hospital of Shiraz University for regular vaccination. The IgG antibody detection against Coxiella burnetii was made by indirect Enzyme-linked Immunosorbent Assay (ELISA), employing phase I and II Coxiella burnetii antigens. A logistic regression model was developed to analyze multiple risk factors associated with seropositivity. An overall seropositivity of 7.7% (n=14) was observed. Prevalence was significantly higher in adult dogs above five years (18.18 %) compared with dogs between 1 and five years (7.86 %) and less than one year (6.17%) (P=0.043). Prevalence was also higher in male dogs (11.21 %) than in female (2.7 %) (P=0.035). There were no significant differences in the prevalence of positive cases and breed, type of housing, type of food and exposure to other farm animals (P>0.05). The results of this study showed the presence of Coxiella burnetii infection among the companion dogs population in Fars province. To our knowledge, this is the first study regarding Q fever in dogs carried out in Iran. In areas like Iran, where human cases of Q fever are not common or remain unreported, the public health implications of Q fever seroprevalence in dogs are quite significant.

Keywords: Coxiella burnetii, dog, Iran, Q fever

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Authors: Mabtho Moreroa-Monyelo, Grace Ijoma, Rosina Nkuna, Tonderayi Matambo

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A moving bed biofilm reactor (MBBR) was used for the bioremediation of high strength chemical oxygen demand (COD) Fisher-Tropsch (FT) wastewater. The aerobic MBBR system was operated over 60 days. For metagenomics profile assessment of the targeted 16S sequence of bacteria involved in the bioremediation of the chemical compounds, sludge samples were collected every second day of operation. Parameters such as pH and COD were measured daily to compare the system efficiency as the changedin microbial diversity progressed. The study revealed that pH was a contributing factor to microbial diversity, which further affected the efficiency of the MBBR system. The highest COD removal rate of 86.4% was achieved at pH 8.3. It was observed that when there was more, A higher bacterial diversity led to an improvement in the reduction of COD. Furthermore, an OTUof 4530 was obtained, which were divided into 12 phyla, 27 classes, 44 orders, 74 families, and 138 genera across all sludge samples from the MBBR. A determination of the relative abundance of microorganisms at phyla level indicates that the most abundant phylum on day it was Firmicutes (50%); thereafter, the most abundant phylum changed toProteobacteria.

Keywords: biodegradation, fischer-tropsch wastewater, metagenomics, moving bed biofilm reactor

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Authors: Muhammad Khalil, Nader Abuelfoutouh, Gasser Abdelal, Adrian Murphy

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Nowadays, the direct effects of lightning to aircrafts are of great importance because of the massive use of composite materials. In comparison with metallic materials, composites present several weaknesses for lightning strike direct effects. Especially, their low electrical and thermal conductivities lead to severe lightning strike damage. The lightning strike direct effects are burning, heating, magnetic force, sparking and arcing. As the problem is complex, we investigated it gradually. A magnetohydrodynamics (MHD) model is developed to simulate the lightning strikes in order to estimate the damages on the composite materials. Then, a coupled thermal-electrical finite element analysis is used to study the interaction between the lightning arc and the composite laminate and to investigate the material degradation.

Keywords: composite structures, lightning multiphysics, magnetohydrodynamic (MHD), coupled thermal-electrical analysis, thermal plasmas.

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Authors: Sara Soltanpour, Adolfo Foriero

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Frost heave is arguably the most problematic adverse phenomenon in cold region areas. Frost heave is a complex process that depends on heat and water transfer. These coupled physical fields generate considerable heave stresses as well as deformations. In the present study, a coupled thermal-hydraulic-mechanical (THM) model using COMSOL Multiphysics in frozen unsaturated soils, such as fine sand, is investigated. Particular attention to the frost heave and temperature distribution, as well as the water migrating during soil freezing, is assessed. The results obtained from the numerical simulations are consistent with the results measured in the full-scale tests conducted by Cold Regions Research and Engineering Laboratory (CRREL).

Keywords: frost heave, numerical simulations, COMSOL software, unsaturated freezing soil

Procedia PDF Downloads 116

Authors: Sarah Devi Silvian, Hanna Shobrina Iqomatul Haq, Fara Cholidatun Nabila, Agustin Krisna Wardani

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Bacteria ability to survive in antibiotic is controlled by the presence of gene that encodes the antibiotic resistance protein. The instability of the antibiotic resistance gene can be observed by exposing the bacteria under the lethal dose of antibiotic. Low concentration of antibiotic can induce mutation, which may take a role in bacterial adaptation through the antibiotic concentration. Lactobacillus casei FNCC 0090 is one of the probiotic bacteria that has an ability to survive in tetracycline by expressing the tetM gene. The aims of this study are to observe the possibilities of mutation happened in L.casei FNCC 0090 by exposing in sub-lethal dose of tetracycline and also observing the instability of the tetM gene by comparing the sequence between the wild type and mutant. L.casei FNCC 0090 has a lethal dose in 60 µg/ml, low concentration is applied to induce the mutation, the range from 10 µg/ml, 15 µg/ml, 30 µg/ml, 45 µg/ml, and 50 µg/ml. L.casei FNCC 0090 is exposed to the low concentration from lowest to the highest concentration to induce the adaptation. Plasmid is isolated from the highest concentration culture which is 50 µg/ml by using modified alkali lysis method with the addition of lysozyme. The tetM gene is isolated by using PCR (Polymerase Chain Reaction) method, then PCR amplicon is purified and sequenced. Sequencing is done on both samples, wild type and mutant. Both sequences are compared and the mutations can be traced in the presence of nucleotides changes. The changing of the nucleotides means that the tetM gene is instable.

Keywords: L. casei FNCC 0090, probiotic, tetM, tetracycline

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Authors: Jihane Saad, Thomas Lendormi, Caroline Le Marechal, Anne-marie Pourcher, Céline Druilhe, Jean-louis Lanoiselle

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Agricultural anaerobic digestion consists in the conversion of animal slurry and manure into biogas and digestate. They need, however, to be treated at 70 ºC during 60 min before anaerobic digestion according to the European regulation (EC n°1069/2009 & EU n°142/2011). The impact of such heat treatment on the outcome of bacteria has been poorly studied up to now. Moreover, a recent study¹ has shown that enterococci and clostridia are still detected despite the application of such thermal treatment, questioning the relevance of this approach for the hygienisation of digestate. The aim of this study is to establish the heat inactivation kinetics of two species of enterococci (Enterococcus faecalis and Enterococcus faecium) and two species of clostridia (Clostridioides difficile and Clostridium novyi as a non-toxic model for Clostridium botulinum of group III). A pure culture of each strain was prepared in a specific sterile medium at concentration of 10⁴ – 10⁷ MPN / mL (Most Probable number), depending on the bacterial species. Bacterial suspensions were then filled in sterilized capillary tubes and placed in a water or oil bath at desired temperature for a specific period of time. Each bacterial suspension was enumerated using a MPN approach, and tests were repeated three times for each temperature/time couple. The inactivation kinetics of the four indicator bacteria is described using the Weibull model and the classical Bigelow model of first-order kinetics. The Weibull model takes biological variation, with respect to thermal inactivation, into account and is basically a statistical model of distribution of inactivation times as the classical first-order approach is a special case of the Weibull model. The heat treatment at 70 ºC / 60 min contributes to a reduction greater than 5 log10 for E. faecium and E. faecalis. However, it results only in a reduction of about 0.7 log10 for C. difficile and an increase of 0.5 log10 for C. novyi. Application of treatments at higher temperatures is required to reach a reduction greater or equal to 3 log10 for C. novyi (such as 30 min / 100 ºC, 13 min / 105 ºC, 3 min / 110 ºC, and 1 min / 115 ºC), raising the question of the relevance of the application of heat treatment at 70 ºC / 60 min for these spore-forming bacteria. To conclude, the heat treatment (70 ºC / 60 min) defined by the European regulation is sufficient to inactivate non-sporulating bacteria. Higher temperatures (> 100 ºC) are required as far as spore-forming bacteria concerns to reach a 3 log10 reduction (sporicidal activity).

Keywords: heat treatment, enterococci, clostridia, inactivation kinetics

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Authors: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri

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Irregular consumption of current antibiotic makes increases of antibiotic resistance between urin pathogens on all worlds. This study selected based on this great community problem. The aim of this study was the biosynthesis of silver nanoparticles from Zataria multiflora extract and then to investigate its antibacterial effect on gram-negative bacilli common in Urinary Tract Infections (UTI) and MDR. The plant used in the present research was Zataria multiflora whose extract was prepared through Soxhlet extraction method. Green synthesis condition of silver nanoparticles was investigated in terms of three parameters including the extract amount, concentration of silver nitrate salt, and temperature. The seizes of nanoparticles were determined by Zetasizer. In order to identify synthesized silver nanoparticles Transmission Electron Microscopy (TEM) and X-ray Diffraction (XRD) methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through biological method different concentrations of silver nanoparticles were studied on 140 cases of Muliple Drug Resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections , for identification of bacteria were used of Polymerase Chain Reaction (PCR) test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were analyzed using SPSS software by nonparametric Kruskal-Wallis and Mann-Whitney tests. Significant results were found about the effects of silver nitrate concentration, different amounts of Zataria multiflora extract, and temperature on nanoparticles; that is, by increasing the concentration of silver nitrate, extract amount, and temperature, the sizes of synthesized nanoparticles declined. However, the effect of above mentioned factors on particles diffusion index was not significant. Based on the TEM results, particles were mainly spherical shape with a diameter range of 25 to 50 nm. The results of XRD Analysis indicated the formation of Nanostructures and Nanocrystals of silver.. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E.coli , Acinetobacter bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125mg/ml and for Acinetobacter bumanii 250mg/ml.Comparing the growth inhibitory effect of chemically synthesized Nanoparticles and biologically synthesized Nanoparticles showed that in the chemical method the highest growth inhibition belonged to the concentration of 62.5 mg /ml. The inhibitory effect on the growth all of bacteria causes of urine infection and MDR was observed and by increasing silver ion concentration in Nanoparticles, antibacterial activity increased. Generally, the biological synthesis can be considered an efficient way not only in making Nanoparticles but also for having anti-bacterial properties. It is more biocompatible and may be possess less toxicity than the Nanoparticles synthesized chemically.

Keywords: biosynthesis, MDR bacteria, silver nanoparticles, UTI

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Authors: Rashida Suleiman, Asamoah Jnr. Boakye, Suleiman Ahmed Ibn Ahmed

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In recent years, there has been an enormous success in discoveries of scientific knowledge in medicine coupled with the advancement of technology. Despite all these successes, diagnoses and treatment of diseases have become complex. MYCIN is a groundbreaking illustration of a clinical decision support system (CDSS), which was developed to assist physicians in the diagnosis and treatment of bacterial infections by providing suggestions for antibiotic regimens. MYCIN was one of the earliest expert systems to demonstrate how CDSSs may assist human decision-making in complicated areas. Relevant databases were searched using google scholar, PubMed and general Google search, which were peculiar to clinical decision support systems. The articles were then screened for a comprehensive overview of the functionality, consultative style and statistical usage of MYCIN, a clinical decision support system. Inferences drawn from the articles showed some usage of MYCIN for problem-based learning among clinicians and students in some countries. Furthermore, the data demonstrated that MYCIN had completed clinical testing at Stanford University Hospital following years of research. The system (MYCIN) was shown to be extremely accurate and effective in diagnosing and treating bacterial infections, and it demonstrated how CDSSs might enhance clinical decision-making in difficult circumstances. Despite the challenges MYCIN presents, the benefits of its usage to clinicians, students and software developers are enormous.

Keywords: clinical decision support system, MYCIN, diagnosis, bacterial infections, support systems

Procedia PDF Downloads 132

Authors: Elham A. Kwildi, Yahia S. Abugnah, Nuri S. Madi

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This study was designed to evaluate the microbiological quality and safety of two types of salads prepared at a catering center affiliated with Libyan Airlines in Tripoli, Libya. Two hundred and twenty-one (221) samples (132 economy-class and 89 first- class) were used in this project which lasted for ten months. Biweekly, microbiological tests were performed which included total plate count (TPC) and total coliforms (TCF), in addition to enumeration and/or detection of some pathogenic bacteria mainly Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella sp, Listeria sp and Vibrio parahaemolyticus parahaemolyticus, By using conventional as well as compact dry methods. Results indicated that TPC of type 1 salad ranged between (<10 – 62 x 103 cfu/gm) and (<10 to 36 x103 cfu/g), while TCF were (<10 – 41 x 103 cfu/gm) and (< 10 to 66 x102 cfu/g) using both methods of detection respectively. On the other hand, TPC of type 2 salad were: (1 × 10 – 52 x 103) and (<10 – 55 x 103 cfu/gm) and in the range of (1 x10 to 45x103 cfu/g), and the (TCF) counts were between (< 10 to 55x103 cfu/g) and (< 10 to 34 x103 cfu/g) using the 1st and the 2nd methods of detection respectively. Also, the pathogens mentioned above were detected in both types of salads, but their levels varied according to the type of salad and the method of detection. The level of Staphylococcus aureus, for instance, was 17.4% using conventional method versus 14.4% using the compact dry method. Similarly, E. coli was 7.6% and 9.8%, while Salmonella sp. recorded the least percentage i.e. 3% and 3.8% with the two mentioned methods respectively. First class salads were also found to contain the same pathogens, but the level of E. coli was relatively higher in this case (14.6% and 16.9%) using conventional and compact dry methods respectively. The second rank came Staphylococcus aureus (13.5%) and (11.2%), followed by Salmonella (6.74%) and 6.70%). The least percentage was for Vibrio parahaemolyticus (4.9%) which was detected in the first class salads only. The other two pathogens Bacillus cereus and Listeria sp. were not detected in either one of the salads. Finally, it is worth mentioning that there was a significant decline in TPC and TCF counts in addition to the disappearance of pathogenic bacteria after the 6-7th month of the study which coincided with the first trial of the HACCP system at the center. The ups and downs in the counts along the early stages of the study reveal that there is a need for some important correction measures including more emphasis on training of the personnel in applying the HACCP system effectively.

Keywords: air travel, vegetable salads, foodborne outbreaks, Libya

Procedia PDF Downloads 315

Authors: Zuliani Mahmood, Thirumulu Ponnuraj Kannan, Yean Yean Chan, Salahddin A. Al-Hudhairy

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Caries is the most common childhood disease which develops due to disturbances in the physiological equilibrium in the dental plaque resulting in demineralization of tooth structures. Plaque and dentine samples were collected from three different tooth surfaces representing caries progression (intact, over carious lesion and dentine) in children with early childhood caries (ECC, n=36). In caries free (CF) children, plaque samples were collected from sound tooth surfaces at baseline and after one year (n=12). The genomic DNA was extracted from all samples and subjected to 16S rRNA PCR amplification. The end products were cloned into pCR®2.1-TOPO® Vector. Five randomly selected positive clones collected from each surface were sent for sequencing. Identification of the bacterial clones was performed using BLAST against GenBank database. In the ECC group, the frequency of Lactobacillus sp. detected was significantly higher in the dentine surface (p = 0.031) than over the cavitated lesion. The highest frequency of bacteria detected in the intact surfaces was Fusobacterium nucleatum subsp. polymorphum (33.3%) while Streptococcus mutans was detected over the carious lesions and dentine surfaces at a frequency of 33.3% and 52.7% respectively. Fusobacterium nucleatum subsp. polymorphum was also found to be highest in the CF group (41.6%). Follow up at the end of one year showed that the frequency of Corynebacterium matruchotii detected was highest in those who remained caries free (16.6%), while Porphyromonas catoniae was highest in those who developed caries (25%). In conclusion, Streptococcus mutans and Porphyromonas catoniae are strongly associated with caries progression, while Lactobacillus sp. is restricted to deep carious lesions. Fusobacterium nucleatum subsp. polymorphum and Corynebacterium matruchotii may play a role in sustaining the healthy equilibrium in the dental plaque. These identified bacteria show promise as potential biomarkers in diagnosis which could help in the management of dental caries in children.

Keywords: early childhood caries, genotypic identification, oral bacteria, 16S rRNA

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Authors: Yves Lacroix, Van Van Than, Didier Léandri, Pierre Liardet

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The western Tombolo of the Giens peninsula in southern France, known as Almanarre beach, is subject to coastal erosion. We are trying to use computer simulation in order to propose solutions to stop this erosion. Our aim was first to determine the main factors for this erosion and successfully apply a coupled hydro-sedimentological numerical model based on observations and measurements that have been performed on the site for decades. We have gathered all available information and data about waves, winds, currents, tides, bathymetry, coastal line, and sediments concerning the site. These have been divided into two sets: one devoted to calibrating a numerical model using Mike 21 software, the other to serve as a reference in order to numerically compare the present situation to what it could be if we implemented different types of underwater constructions. This paper presents the first part of the study: selecting and melting different sources into a coherent data basis, identifying the main erosion factors, and calibrating the coupled software model against the selected reference period. Our results bring calibration of the numerical model with good fitting coefficients. They also show that the winter South-Western storm events conjugated to depressive weather conditions constitute a major factor of erosion, mainly due to wave impact in the northern part of the Almanarre beach. Together, current and wind impact is shown negligible.

Keywords: Almanarre beach, coastal erosion, hydro-sedimentological, numerical model

Procedia PDF Downloads 373

Authors: Sendi Zohra, Belhadjsalah Hedi, Labergere Carl, Saanouni Khemais

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The modeling of mechanical problems including both material and geometric nonlinearities with Finite Element Method (FEM) remains challenging. Meshless methods offer special properties to get rid of well-known drawbacks of the FEM. The main objective of Meshless Methods is to eliminate the difficulty of meshing and remeshing the entire structure by simply insertion or deletion of nodes, and alleviate other problems associated with the FEM, such as element distortion, locking and others. In this study, a robust numerical implementation of an Element Free Galerkin Method for an elastoplastic coupled to damage problem is presented. Several results issued from the numerical simulations by a DynamicExplicit resolution scheme are analyzed and critically compared with Element Finite Method results. Finally, different numerical examples are carried out to demonstrate the efficiency of this method.

Keywords: damage, dynamic explicit, elastoplasticity, isotropic hardening, meshless

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Authors: Rajat Raj, Rohini S. Hallikar

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The ever-increasing integration of renewable energy sources in the power grid necessitates the development of efficient and reliable microgrid systems. Photovoltaic (PV) systems coupled with energy storage technologies, such as batteries, offer promising solutions for sustainable and resilient power generation. This paper proposes an adaptive power control topology for a PV-battery microgrid system, aiming to optimize the utilization of available solar energy and enhance the overall system performance. In order to provide a smooth transition between the OFF-GRID and ON-GRID modes of operation with proportionate power sharing, a self-adaptive control method for a microgrid is proposed. Three different modes of operation are discussed in this paper, i.e., GRID connected, the transition between Grid-connected and Islanded State, and changing the irradiance of PVs and doing the transitioning. The simulation results show total harmonic distortion to be 0.08, 1.43 and 2.17 for distribution generation-1 and 4.22,3.92 and 2.10 for distribution generation-2 in the three modes, respectively which helps to maintain good power quality. The simulation results demonstrate the superiority of the adaptive power control topology in terms of maximizing renewable energy utilization, improving system stability and ensuring a seamless transition between grid-connected and islanded modes.

Keywords: islanded modes, microgrids, photo voltaic, total harmonic distortion

Procedia PDF Downloads 161