Search results for: Bacillus strain IIIJ3-1

Authors: A. S. Ibrahim Mamane, S. Giljean, M.-J. Pac, G. L’Hostis

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Fiber reinforced composite laminates are particularly subject to residual stresses due to their heterogeneity and the complex chemical, mechanical and thermal mechanisms that occur during their processing. Residual stresses are now well known to cause damage accumulation, shape instability, and behavior disturbance in composite parts. Many works exist in the literature on techniques for minimizing residual stresses in thermosetting and thermoplastic composites mainly. To study in-depth the influence of processing mechanisms on the formation of residual stresses and to minimize them by establishing a reliable correlation, it is essential to be able to measure very precisely the profile of residual stresses in the composite. Residual stresses are important data to consider when sizing composite parts and predicting their behavior. The incremental hole drilling is very effective in measuring the gradient of residual stresses in composite laminates. This method is semi-destructive and consists of drilling incrementally a hole through the thickness of the material and measuring relaxation strains around the hole for each increment using three strain gauges. These strains are then converted into residual stresses using a matrix of coefficients. These coefficients, called calibration coefficients, depending on the diameter of the hole and the dimensions of the gauges used. The reliability of the incremental hole drilling depends on the accuracy with which the calibration coefficients are determined. These coefficients are calculated using a finite element model. The samples’ features and the experimental conditions must be considered in the simulation. Any mismatch can lead to inadequate calibration coefficients, thus introducing errors on residual stresses. Several calibration coefficient correction methods exist for isotropic material, but there is a lack of information on this subject concerning composite laminates. In this work, a Python program was developed to automatically generate the adequate finite element model. This model allowed us to perform a parametric study to assess the influence of experimental errors on the calibration coefficients. The results highlighted the sensitivity of the calibration coefficients to the considered errors and gave an order of magnitude of the precisions required on the experimental device to have reliable measurements. On the basis of these results, improvements were proposed on the experimental device. Furthermore, a numerical method was proposed to correct the calibration coefficients for different types of materials, including thick composite parts for which the analytical approach is too complex. This method consists of taking into account the experimental errors in the simulation. Accurate measurement of the experimental errors (such as eccentricity of the hole, angular deviation of the gauges from their theoretical position, or errors on increment depth) is therefore necessary. The aim is to determine more precisely the residual stresses and to expand the validity domain of the incremental hole drilling technique.

Keywords: fiber reinforced composites, finite element simulation, incremental hole drilling method, numerical correction of the calibration coefficients, residual stresses

Procedia PDF Downloads 108

Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

Procedia PDF Downloads 244

Authors: Nezmin Kayedpour, Arash E. Samani, Siavash Asiaban, Jeroen M. De Kooning, Lieven Vandevelde, Guillaume Crevecoeur

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The growing adoption of renewable energy sources, such as wind power, in electricity generation is a significant step towards a sustainable and decarbonized future. However, the inherent intermittency and uncertainty of wind resources pose challenges to the reliable and stable operation of power grids. To address this, hydrogen storage systems have emerged as a promising and versatile technology to support grid balancing services in wind energy conversion systems. In this study, we propose a supplementary control design that enhances the performance of the hydrogen storage system by integrating wind turbine (WT) pitch and torque control systems. These control strategies aim to optimize the hydrogen production process, ensuring efficient utilization of wind energy while complying with grid requirements. The wind turbine pitch control system plays a crucial role in managing the turbine's aerodynamic performance. By adjusting the blade pitch angle, the turbine's rotational speed and power output can be regulated. Our proposed control design dynamically coordinates the pitch angle to match the wind turbine's power output with the optimal hydrogen production rate. This ensures that the electrolyzer receives a steady and optimal power supply, avoiding unnecessary strain on the system during high wind speeds and maximizing hydrogen production during low wind speeds. Moreover, the wind turbine torque control system is incorporated to facilitate efficient operation at varying wind speeds. The torque control system optimizes the energy capture from the wind while limiting mechanical stress on the turbine components. By harmonizing the torque control with hydrogen production requirements, the system maintains stable wind turbine operation, thereby enhancing the overall energy-to-hydrogen conversion efficiency. To enable grid-friendly operation, we introduce a cascaded controller that regulates the electrolyzer's electrical power-current in accordance with grid requirements. This controller ensures that the hydrogen production rate can be dynamically adjusted based on real-time grid demands, supporting grid balancing services effectively. By maintaining a close relationship between the wind turbine's power output and the electrolyzer's current, the hydrogen storage system can respond rapidly to grid fluctuations and contribute to enhanced grid stability. In this paper, we present a comprehensive analysis of the proposed supplementary control design's impact on the overall performance of the hydrogen storage system in wind energy conversion systems. Through detailed simulations and case studies, we assess the system's ability to provide grid balancing services, maximize wind energy utilization, and reduce greenhouse gas emissions.

Keywords: active power control, electrolyzer, grid balancing services, wind energy conversion systems

Procedia PDF Downloads 55

Authors: Ružica Tomičić, Zorica Tomičić, Peter Raspor

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An abundant growth of unwanted yeasts in food processing plants can lead to problems in quality and safety with significant financial losses. Candida and Pichia are the genera mainly involved in spoilage of products in the food and beverage industry. These contaminating microorganisms can form biofilms on food contact surfaces, being difficult to eradicate, increasing the probability of microbial survival and further dissemination during food processing. It is well known that biofilms are more resistant to antimicrobial agents compared to planktonic cells and this makes them difficult to eliminate. Among the strategies used to overcome resistance to antifungal drugs and preservatives, the use of natural substances such as plant extracts has shown particular promise, and many natural substances have been found to exhibit antifungal properties. This study aimed to investigated the impact of growth medium (Malt Extract broth (MEB) or Yeast Peptone Dextrose (YPD) broth) and temperatures (7°C, 37°C, 43°C for Candida strains and 7°C, 27°C, 32°C for Pichia strains) on the adhesion of Candida spp. and Pichia spp. to stainless steel (AISI 304) discs with different degrees of surface roughness (Ra = 25.20 – 961.9 nm), a material commonly used in the food industry. We also evaluated the antifungal and antiadhesion activity of plant extracts such as Humulus lupulus, Alpinia katsumadai and Evodia rutaecarpa against C. albicans, C glabrata and P. membranifaciens and investigated whether these plant extracts can interfere with biofilm formation. The adhesion was assessed by the crystal violet staining method, while the broth microdilution method CLSI M27-A3 was used to determine the minimum inhibitory concentration (MIC) of plant extracts. Our results indicated that the nutrient content of the medium significantly influenced the amount of adhered cells of the tested yeasts. The growth medium which resulted in a higher adhesion of C. albicans and C. glabrata was MEB, while for C. parapsilosis and C. krusei was YPD. In the case of P. pijperi and P. membranifaciens, YPD broth was more effective in promoting adhesion than MEB. Regarding the effect of temperature, C. albicans strain adhered to stainless steel surfaces in significantly higher level at a temperature of 43°C, while on the other hand C. glabrata, C. parapsilosis and C. krusei showed a different behavior with significantly higher adhesion at 37°C than at 7°C and 43°C. Further, the adherence ability of Pichia strains was highest at 27°C. Based on the MIC values, all plant extracts exerted significant antifungal effects with MIC values ranged from 100 to 400 μg/mL. It was observed that biofilm of C. glabrata were more resistance to plant extracts as compared to C. albicans. However, extracts of A. katsumadai and E. rutaecarpa promoted the growth and development of the preformed biofilm of P. membranifaciens. Thus, the knowledge of how these microorganisms adhere and which factors affect this phenomenon is of great importance in order to avoid their colonization on food contact surfaces.

Keywords: adhesion, Candida spp., Pichia spp., plant extracts

Procedia PDF Downloads 175

Authors: Andrey A. Tyuftin, Rachael Reid, Paula Bourke, Patrick J. Cullen, Seamus Fanning, Paul Whyte, Declan Bolton , Joe P. Kerry

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One of the primary spoilage issues associated with vacuum-packed beef products is blown pack spoilage (BPS) caused by the psychrophilic spore-forming strain of Clostridium spp. Spores derived from this organism can be activated after heat-shrinking (eg. 90°C for 3 seconds). To date, research into the control of Clostridium spp in beef packaging is limited. Active packaging in the form of antimicrobially-active coatings may be one approach to its control. Antimicrobial compounds may be incorporated into packaging films or coated onto the internal surfaces of packaging films using a carrier matrix. Three naturally-sourced, commercially-available antimicrobials, namely; Auranta FV (AFV) (bitter oranges extract) from Envirotech Innovative Products Ltd, Ireland; Inbac-MDA (IMDA) from Chemital LLC, Spain, mixture of different organic acids and sodium octanoate (SO) from Sigma-Aldrich, UK, were added into gelatin solutions at 2 concentrations: 2.5 and 3.5 times their minimum inhibition concentration (MIC) against Clostridium estertheticum (DSMZ 8809). These gelatin solutions were coated onto the internal polyethylene layer of cold plasma treated, heat-shrinkable laminates conventionally used for meat packaging applications. Atmospheric plasma was used in order to enhance adhesion between packaging films and gelatin coatings. Pouches were formed from these coated packaging materials, and beef cuts which had been inoculated with C. estertheticum were vacuum packaged. Inoculated beef was vacuum packaged without employing active films and this treatment served as the control. All pouches were heat-sealed and then heat-shrunk at 90°C for 3 seconds and incubated at 2°C for 100 days. During this storage period, packs were monitored for the indicators of blown pack spoilage as follows; gas bubbles in drip, loss of vacuum (onset of BPS), blown, the presence of sufficient gas inside the packs to produce pack distension and tightly stretched, “overblown” packs/ packs leaking. Following storage and assessment of indicator date, it was concluded that AFV- and SO-containing packaging inhibited the growth of C. estertheticum, significantly delaying the blown pack spoilage of beef primals. IMDA did not inhibit the growth of C. estertheticum. This may be attributed to differences in release rates and possible reactions with gelatin. Overall, active films were successfully produced following plasma surface treatment, and experimental data demonstrated clearly that the use of antimicrobially-active films could significantly prolong the storage stability of beef primals through the effective control of BPS.

Keywords: active packaging, blown pack spoilage, Clostridium, antimicrobials, edible coatings, food packaging, gelatin films, meat science

Procedia PDF Downloads 239

Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

Procedia PDF Downloads 213

Authors: Dina Kon Mushid, Kabutakapua Kakanda, Dibu Dave Mbako

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The failure of material usually involves elastoplastic deformation and fracturing. Continuum mechanics can effectively deal with plastic deformation by using a yield function and the flow rule. At the same time, it has some limitations in dealing with the fracture problem since it is a theory based on the continuous field hypothesis. The lattice model can simulate the fracture problem very well, but it is inadequate for dealing with plastic deformation. Based on the discretized virtual internal bond model (DVIB), this paper proposes a lattice model that can account for plasticity. DVIB is a lattice method that considers material to comprise bond cells. Each bond cell may have any geometry with a finite number of bonds. The two-body or multi-body potential can characterize the strain energy of a bond cell. The two-body potential leads to the fixed Poisson ratio, while the multi-body potential can overcome the limitation of the fixed Poisson ratio. In the present paper, the modified Stillinger-Weber (SW), a multi-body potential, is employed to characterize the bond cell energy. The SW potential is composed of two parts. One part is the two-body potential that describes the interatomic interactions between particles. Another is the three-body potential that represents the bond angle interactions between particles. Because the SW interaction can represent the bond stretch and bond angle contribution, the SW potential-based DVIB (SW-DVIB) can represent the various Poisson ratios. To embed the plasticity in the SW-DVIB, the plasticity is considered in the two-body part of the SW potential. It is done by reducing the bond stiffness to a lower level once the bond reaches the yielding point. While before the bond reaches the yielding point, the bond is elastic. When the bond deformation exceeds the yielding point, the bond stiffness is softened to a lower value. When unloaded, irreversible deformation occurs. With the bond length increasing to a critical value, termed the failure bond length, the bond fails. The critical failure bond length is related to the cell size and the macro fracture energy. By this means, the fracture energy is conserved so that the cell size sensitivity problem is relieved to a great extent. In addition, the plasticity and the fracture are also unified at the bond level. To make the DVIB able to simulate different Poisson ratios, the three-body part of the SW potential is kept elasto-brittle. The bond angle can bear the moment before the bond angle increment is smaller than a critical value. By this method, the SW-DVIB can simulate the plastic deformation and the fracturing process of material with various Poisson ratios. The elastoplastic SW-DVIB is used to simulate the plastic deformation of a material, the plastic fracturing process, and the tunnel plastic deformation. It has been shown that the current SW-DVIB method is straightforward in simulating both elastoplastic deformation and plastic fracture.

Keywords: lattice model, discretized virtual internal bond, elastoplastic deformation, fracture, modified stillinger-weber potential

Procedia PDF Downloads 71

Authors: Sonia Kudlacik-Kramarczyk, Anna Drabczyk, Dagmara Malina, Bozena Tyliszczak, Agnieszka Sobczak-Kupiec

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Introduction: In the preparation of polymer materials, compounds of natural origin are currently gaining more and more interest. This is particularly noticeable in the case of synthesis of materials considered for biomedical use. Then, selected material has to meet many requirements. It should be characterized by non-toxicity, biodegradability and biocompatibility. Therefore special attention is directed to substances such as polysaccharides, proteins or substances that are the basic building components of proteins, i.e. amino acids. These compounds may be crosslinked with other reagents that leads to the preparation of polymer matrices. Such amino acids as e.g. cysteine or histidine. On the other hand, previously mentioned requirements may be met by polymers obtained as a result of biosynthesis, e.g. polyhydroxybutyrate. This polymer belongs to the group of aliphatic polyesters that is synthesized by microorganisms (selected strain of bacteria) under specific conditions. It is possible to modify matrices based on given polymer with substances of various origin. Such a modification may result in the change of their properties or/and in providing the material with new features desirable in viewpoint of specific application. Described materials are synthesized using UV radiation. Process of photopolymerization is fast, waste-free and enables to obtain final products with favorable properties. Methodology: Polymer matrices have been prepared by means of photopolymerization. First step involved the preparation of solutions of particular reagents and mixing them in the appropriate ratio. Next, crosslinking agent and photoinitiator have been added to the reaction mixture and the whole was poured into the Petri dish and treated with UV radiation. After the synthesis, polymer samples were dried at room temperature and subjected to the numerous analyses aimed at the determining their physicochemical properties. Firstly, sorption properties of obtained polymer matrices have been determined. Next, mechanical properties have been characterized, i.e. tensile strength. The ability to deformation under applied stress of all prepared polymer matrices has been checked. Such a property is important in viewpoint of the application of analyzed materials e.g. as wound dressings. Wound dressings have to be elastic because depending on the location of the wound and its mobility, such a dressing has to adhere properly to the wound. Furthermore, considering the use of the materials for biomedical purposes it is essential to determine its behavior in environments simulating these ones occurring in human body. Therefore incubation studies using selected liquids have also been conducted. Conclusions: As a result of photopolymerization process, polymer matrices based on natural compounds have been prepared. These exhibited favorable mechanical properties and swelling ability. Moreover, biocompatibility in relation to simulated body fluids has been stated. Therefore it can be concluded that analyzed polymer matrices constitute an interesting materials that may be considered for biomedical use and may be subjected to the further more advanced analyses using specific cell lines.

Keywords: photopolymerization, polymer matrices, simulated body fluids, swelling properties

Procedia PDF Downloads 100

Authors: Barbara Urasinska-Wojcik, Neil Chilton, Peter Todd, Christopher Elsworthy, Gregory J. Gibbons

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The recent increase in the application of Additive Manufacturing (AM) of products has resulted in new demands on capability. The ability to integrate both form and function within printed objects is the next frontier in the 3D printing area. To move beyond prototyping into low volume production, we demonstrate a UK-designed and built AM hybrid system that combines polymer based structural deposition with digital deposition of electrically conductive elements. This hybrid manufacturing system is based on a multi-planar build approach to improve on many of the limitations associated with AM, such as poor surface finish, low geometric tolerance, and poor robustness. Specifically, the approach involves a multi-planar Material Extrusion (ME) process in which separated build stations with up to 5 axes of motion replace traditional horizontally-sliced layer modeling. The construction of multi-material architectures also involved using multiple print systems in order to combine both ME and digital deposition of conductive material. To demonstrate multi-material 3D printing, three thermoplastics, acrylonitrile butadiene styrene (ABS), polyamide 6,6/6 copolymers (CoPA) and polyamide 12 (PA) were used to print specimens, on top of which our high viscosity Ag-particulate ink was printed in a non-contact process, during which drop characteristics such as shape, velocity, and volume were assessed using a drop watching system. Spectroscopic analysis of these 3D printed materials in the IR region helped to determine the optimum in-situ curing system for implementation into the AM system to achieve improved adhesion and surface refinement. Thermal Analyses were performed to determine the printed materials glass transition temperature (Tg), stability and degradation behavior to find the optimum annealing conditions post printing. Electrical analysis of printed conductive tracks on polymer surfaces during mechanical testing (static tensile and 3-point bending and dynamic fatigue) was performed to assess the robustness of the electrical circuits. The tracks on CoPA, ABS, and PA exhibited low electrical resistance, and in case of PA resistance values of tracks remained unchanged across hundreds of repeated tensile cycles up to 0.5% strain amplitude. Our developed AM printer has the ability to fabricate fully functional objects in one build, including complex electronics. It enables product designers and manufacturers to produce functional saleable electronic products from a small format modular platform. It will make 3D printing better, faster and stronger.

Keywords: additive manufacturing, conductive tracks, hybrid 3D printer, integrated manufacture

Procedia PDF Downloads 143

Authors: O. Bilbao, I. Loizaga, F. A. Girot, A. Torregaray

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The current energetical situation and the high competitiveness in industrial sectors as the automotive one have become the development of new manufacturing processes with less energy and raw material consumption a real necessity. As consequence, new forming processes related with high temperature hot forging in closed dies have emerged in the last years as new solutions to expand the possibilities of hot forging and iron casting in the automotive industry. These technologies are mid-way between hot forging and semi-solid metal processes, working at temperatures higher than the hot forging but below the solidus temperature or the semi solid range, where no liquid phase is expected. This represents an advantage comparing with semi-solid forming processes as thixoforging, by the reason that no so high temperatures need to be reached in the case of high melting point alloys as steels, reducing the manufacturing costs and the difficulties associated to semi-solid processing of them. Comparing with hot forging, this kind of technologies allow the production of parts with as forged properties and more complex and near-net shapes (thinner sidewalls), enhancing the possibility of designing lightweight components. From the process viewpoint, the forging forces are significantly decreased, and a significant reduction of the raw material, energy consumption, and the forging steps have been demonstrated. Despite the mentioned advantages, from the material behavior point of view, the expansion of these technologies has shown the necessity of developing new material flow behavior models in the process working temperature range to make the simulation or the prediction of these new forming processes feasible. Moreover, the knowledge of the material flow behavior at the working temperature range also allows the design of the new closed dies concept required. In this work, the flow behavior characterization in the mentioned temperature range of the widely used in automotive commercial components 42CrMo4 steel has been studied. For that, hot compression tests have been carried out in a thermomechanical tester in a temperature range that covers the material behavior from the hot forging until the NDT (Nil Ductility Temperature) temperature (1250 ºC, 1275 ºC, 1300 ºC, 1325 ºC, 1350ºC, and 1375 ºC). As for the strain rates, three different orders of magnitudes have been considered (0,1 s-1, 1s-1, and 10s-1). Then, results obtained from the hot compression tests have been treated in order to adapt or re-write the Spittel model, widely used in automotive commercial softwares as FORGE® that restrict the current existing models up to 1250ºC. Finally, the obtained new flow behavior model has been validated by the process simulation in a commercial automotive component and the comparison of the results of the simulation with the already made experimental tests in a laboratory cellule of the new technology. So as a conclusion of the study, a new flow behavior model for the 42CrMo4 steel in the new working temperature range and the new process simulation in its application in automotive commercial components has been achieved and will be shown.

Keywords: 42CrMo4 high temperature flow behavior, high temperature hot forging in closed dies, simulation of automotive commercial components, spittel flow behavior model

Procedia PDF Downloads 96

Authors: M. Hosseini, N. Ghorbani Amirabad, M. Zhian

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Seismic design codes accept structural and nonstructural damages after the sever earthquakes (provided that the building is prevented from collapse), so that in many cases demolishing and reconstruction of the building is inevitable, and this is usually very difficult, costly and time consuming. Therefore, designing and constructing of buildings in such a way that they can be easily repaired after earthquakes, even major ones, is quite desired. For this purpose giving the possibility of rocking or see-saw motion to the building structure, partially or as a whole, has been used by some researchers in recent decade .the central support which has a main role in creating the possibility of see-saw motion in the building’s structural system. In this paper, paying more attention to the key role of the central fuse and support, an innovative energy dissipater which can act as the central fuse and support of the building with seesaw motion is introduced, and the process of reaching an optimal geometry for that by using finite element analysis is presented. Several geometric shapes were considered for the proposed central fuse and support. In each case the hysteresis moment rotation behavior of the considered fuse were obtained under simultaneous effect of vertical and horizontal loads, by nonlinear finite element analyses. To find the optimal geometric shape, the maximum plastic strain value in the fuse body was considered as the main parameter. The rotational stiffness of the fuse under the effect of acting moments is another important parameter for finding the optimum shape. The proposed fuse and support can be called Yielding Curved Bars and Clipped Hemisphere Core (YCB&CHC or more briefly YCB) energy dissipater. Based on extensive nonlinear finite element analyses it was found out the using rectangular section for the curved bars gives more reliable results. Then, the YCB energy dissipater with the optimal shape was used in a structural model of a 12 story regular building as its central fuse and support to give it the possibility of seesaw motion, and its seismic responses were compared to those of a the building in the fixed based conditions, subjected to three-components acceleration of several selected earthquakes including Loma Prieta, Northridge, and Park Field. In building with see-saw motion some simple yielding-plate energy dissipaters were also used under circumferential columns.The results indicated that equipping the buildings with central and circumferential fuses result in remarkable reduction of seismic responses of the building, including the base shear, inter story drift, and roof acceleration. In fact by using the proposed technique the plastic deformations are concentrated in the fuses in the lowest story of the building, so that the main body of the building structure remains basically elastic, and therefore, the building can be easily repaired after earthquake.

Keywords: rocking mechanism, see-saw motion, finite element analysis, hysteretic behavior

Procedia PDF Downloads 386

Authors: Sundru Manjulata Devi, Prakash M. Halami

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The immemorial use of fermented foods from vegetables, dairy and other biological sources are of great demand in India because of their health benefits. However, the diversity of Lactobacillus plantarum group (LPG) of vegetable origin has not been revealed yet, particularly with reference to their probiotic functionalities. In the present study, the different species of probiotic Lactobacillus plantarum group (LPG) i.e., L. plantarum subsp. plantarum MTCC 5422 (from fermented cereals), L. plantarum subsp. argentoratensis FG16 (from fermented bamboo shoot) and L. paraplantarum MTCC 9483 (from fermented gundruk) (as characterized by multiplex recA PCR assay) were considered to investigate their relative expression of MUB domains of mub gene (mucin binding protein) by Real time PCR. Initially, the allelic variation in the mub gene was assessed and found to encode three different variants (Type I, II and III). All the three types had 8, 9 and 10 MUB domains respectively (as analysed by Pfam database) and were found to be responsible for adhesion of bacteria to the host intestinal epithelial cells. These domains either get inserted or deleted during speciation or evolutionary events and lead to divergence. The reverse transcriptase qPCR analysis with mubLPF1+R1 primer pair supported variation in amplicon sizes with 300, 500 and 700 bp among different LPG strains. The relative expression of these MUB domains significantly unregulated in the presence of 1% mucin in overnight grown cultures. Simultaneously, the mub gene expressed efficiently by 7 fold in the culture L. paraplantarum MTCC 9483 with 10 MUB domains. An increase in the expression levels for L. plantarum subsp. plantarum MTCC 5422 and L. plantarum subsp. argentoratensis FG16 (MCC 2974) with 9 and 8 repetitive domains was around 4 and 2 fold, respectively. The detection and expression of an integrase (int) gene in the upstream region of mub gene reveals the excision and integration of these repetitive domains. Concurrently, an in vitro adhesion assay to mucin and exclusion of pathogens (such as Listeria monocytogenes and Micrococcus leuteus) was investigated and observed that the L. paraplantarum MTCC 9483 with more adhesion domains has more ability to adhere to mucin and inhibited the growth of pathogens. The production and expression of plantaricin-like bacteriocin (plnNC8 type) in MTCC 9483 suggests the pathogen inhibition. Hence, the expression of MUB domains can act as potential biomarkers in the screening of a novel probiotic LPG strain with adherence property. The present study provides a platform for an easy, rapid, less time consuming, low-cost methodology for the detection of potential probiotic bacteria. It was known that the traditional practices followed in the preparation of fermented bamboo shoots/gundruk/cereals of Indian foods contain different kinds of neutraceuticals for functional food and novel compounds with health promoting factors. In future, a detailed study of these food products can add more nutritive value, consumption and suitable for commercialization.

Keywords: adhesion gene, fermented foods, MUB domains, probiotics

Procedia PDF Downloads 240

Authors: Marcel Meinhardt, Manfred Keuser, Thomas Braml

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Cracking of reinforced concrete is a complex phenomenon induced by direct loads or restraints affecting reinforced concrete structures as soon as the tensile strength of the concrete is exceeded. Hence it is important to predict where cracks will be located and how they will propagate. The bond theory and the crack formulas in the actual design codes, for example, DIN EN 1992-1-1, are all based on the assumption that the reinforcement bars are embedded in homogeneous concrete without taking into account the influence of transverse reinforcement and the real stress situation. However, it can often be observed that real structures such as walls, slabs or beams show a crack spacing that is orientated to the transverse reinforcement bars or to the stirrups. In most Finite Element Analysis studies, the smeared crack approach is used for crack prediction. The disadvantage of this model is that the typical strain localization of a crack on element level can’t be seen. The crack propagation in concrete is a discontinuous process characterized by different factors such as the initial random distribution of defects or the scatter of material properties. Such behavior presupposes the elaboration of adequate models and methods of simulation because traditional mechanical approaches deal mainly with average material parameters. This paper concerned with the modelling of the initiation and the propagation of cracks in reinforced concrete structures considering the influence of transverse reinforcement and the real stress distribution in reinforced concrete (R/C) beams/plates in bending action. Therefore, a parameter study was carried out to investigate: (I) the influence of the transversal reinforcement to the stress distribution in concrete in bending mode and (II) the crack initiation in dependence of the diameter and distance of the transversal reinforcement to each other. The numerical investigations on the crack initiation and propagation were carried out with a 2D reinforced concrete structure subjected to quasi static loading and given boundary conditions. To model the uncertainty in the tensile strength of concrete in the Finite Element Analysis correlated normally and lognormally distributed random filed with different correlation lengths were generated. The paper also presents and discuss different methods to generate random fields, e.g. the Covariance Matrix Decomposition Method. For all computations, a plastic constitutive law with softening was used to model the crack initiation and the damage of the concrete in tension. It was found that the distributions of crack spacing and crack widths are highly dependent of the used random field. These distributions are validated to experimental studies on R/C panels which were carried out at the Laboratory for Structural Engineering at the University of the German Armed Forces in Munich. Also, a recommendation for parameters of the random field for realistic modelling the uncertainty of the tensile strength is given. The aim of this research was to show a method in which the localization of strains and cracks as well as the influence of transverse reinforcement on the crack initiation and propagation in Finite Element Analysis can be seen.

Keywords: crack initiation, crack modelling, crack propagation, cracks, numerical simulation, random fields, reinforced concrete, stochastic

Procedia PDF Downloads 119

Authors: Garima Singh

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Objectives: Behavioral Crisis Center (BCC) is a community solution to a community problem. There has been an exponential increase in the incidence and prevalence of mental health crises around the world. The effects of the crisis negatively impact our patients and their families and strain the law enforcement and emergency room. The goal of the multi-disciplinary care model is to break the crisis cycle and provide 24-7 rapid access to an acre and crisis stabilization. We initiated our first BCC care center in 2020 in the midst of the COVID pandemic and have seen a remarkable improvement in patient ‘care and positive financial outcome. Background: Mental illnesses are common in the United States. Nearly one in five U.S. adults live with a mental illness (52.9 million in 2020). This number represented 21.0% of all U.S. adults. To address some of these challenges and help our community, In May 2020, we opened our first Behavioral crisis center (BCC). Since then, we have served more than 2500 patients and is the first southwest Missouri’s first 24/7 facility for crisis–level behavioral health and substance use needs. It has been proven to be a more effective place than emergency departments, jails, or local law enforcement. Methods: BCC was started in 2020 to serve the unmet need of the community and provide access to behavioral health and substance use services identified in the community. Funding was possible with significant investment from the county and Missouri Foundation for Health, with contributions from medical partners. It is a multi-disciplinary care center consisting of Physicians, nurse practitioners, nurses, behavioral technicians, peer support specialists, clinical intake specialists, and clinical coordinators and hospitality specialists. The center provides services including psychiatry care, outpatient therapy, community support services, primary care, peer support and engagement. It is connected to a residential treatment facility for substance use treatment for continuity of care and bridging the gap, which has resulted in the completion of treatment and better outcomes. Results: BCC has proven to be a great resource to the community and the Missouri Health Coalition is providing funding to replicate the model in other regions and work on a similar model for children and adolescents. Overall, 29% of the patients seen at BCC are stabilized and discharged with outpatient care. 50% needed acute stabilization in a hospital setting and 21% required long-term admission, mostly for substance use treatment. The local emergency room had a 42% reduction in behavioral health encounters compared to the previous 3 years. Also, by a quick transfer to BCC, the average stay in ER was reduced by 10 hours and time to follow up behavioral health assessment decreased by an average of 4 hours. Uninsured patients are also provided Medicaid application assistance which has benefited 55% of individuals receiving care at BCC. Conclusions: BCC is impacting community health and improving access to quality care and substance use treatment. It is a great investment for our patients and families.

Keywords: BCC, behvaioral health, community health care, addiction treatment

Procedia PDF Downloads 50

Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

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Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

Procedia PDF Downloads 245

Authors: Ashok Kumar, Shiv Brat Singh, Kalyan Kumar Ray

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There is growing awareness to design steels against fatigue damage Ferrite martensite dual-phase steels are known to exhibit favourable mechanical properties like good strength, ductility, toughness, continuous yielding, and high work hardening rate. However, dual-phase steels containing bainite as second phase are potential alternatives for ferrite martensite steels for certain applications where good fatigue property is required. Fatigue properties of dual phase steels are popularly assessed by the nature of variation of crack growth rate (da/dN) with stress intensity factor range (∆K), and the magnitude of fatigue threshold (∆Kth) for long cracks. There exists an increased emphasis to understand not only the long crack fatigue behavior but also short crack growth behavior of ferrite bainite dual phase steels. The major objective of this report is to examine the influence of microstructures on the short and long crack growth behavior of a series of developed dual-phase steels with varying amounts of bainite and. Three low carbon steels containing Nb, Cr and Mo as microalloying elements steels were selected for making ferrite-bainite dual-phase microstructures by suitable heat treatments. The heat treatment consisted of austenitizing the steel at 1100°C for 20 min, cooling at different rates in air prior to soaking these in a salt bath at 500°C for one hour, and finally quenching in water. Tensile tests were carried out on 25 mm gauge length specimens with 5 mm diameter using nominal strain rate 0.6x10⁻³ s⁻¹ at room temperature. Fatigue crack growth studies were made on a recently developed specimen configuration using a rotating bending machine. The crack growth was monitored by interrupting the test and observing the specimens under an optical microscope connected to an Image analyzer. The estimated crack lengths (a) at varying number of cycles (N) in different fatigue experiments were analyzed to obtain log da/dN vs. log °∆K curves for determining ∆Kthsc. The microstructural features of these steels have been characterized and their influence on the near threshold crack growth has been examined. This investigation, in brief, involves (i) the estimation of ∆Kthsc and (ii) the examination of the influence of microstructure on short and long crack fatigue threshold. The maximum fatigue threshold values obtained from short crack growth experiments on various specimens of dual-phase steels containing different amounts of bainite are found to increase with increasing bainite content in all the investigated steels. The variations of fatigue behavior of the selected steel samples have been explained with the consideration of varying amounts of the constituent phases and their interactions with the generated microstructures during cyclic loading. Quantitative estimation of the different types of fatigue crack paths indicates that the propensity of a crack to pass through the interfaces depends on the relative amount of the microstructural constituents. The fatigue crack path is found to be predominantly intra-granular except for the ones containing > 70% bainite in which it is predominantly inter-granular.

Keywords: bainite, dual phase steel, fatigue crack growth rate, long crack fatigue threshold, short crack fatigue threshold

Procedia PDF Downloads 185

Authors: M. N. Nwodo, Y. P. Cheng, N. H. Minh

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In the process of recovering oil in weak sandstone formations, the strength of sandstones around the wellbore is weakened due to the increase of effective stress/load from the completion activities around the cavity. The weakened and de-bonded sandstone may be eroded away by the produced fluid, which is termed sand production. It is one of the major trending subjects in the petroleum industry because of its significant negative impacts, as well as some observed positive impacts. For efficient sand management therefore, there has been need for a reliable study tool to understand the mechanism of sanding. One method of studying sand production is the use of the widely recognized Discrete Element Method (DEM), Particle Flow Code (PFC3D) which represents sands as granular individual elements bonded together at contact points. However, there is limited knowledge of the particle-scale behavior of the weak sandstone, and the parameters that affect sanding. This paper aims to investigate the reliability of using PFC3D and a simple Darcy flow in understanding the sand production behavior of a weak sandstone. An isotropic tri-axial test on a weak oil sandstone sample was first simulated at a confining stress of 1MPa to calibrate and validate the parallel bond models of PFC3D using a 10m height and 10m diameter solid cylindrical model. The effect of the confining stress on the number of bonds failure was studied using this cylindrical model. With the calibrated data and sample material properties obtained from the tri-axial test, simulations without and with fluid flow were carried out to check on the effect of Darcy flow on bonds failure using the same model geometry. The fluid flow network comprised of every four particles connected with tetrahedral flow pipes with a central pore or flow domain. Parametric studies included the effects of confining stress, and fluid pressure; as well as validating flow rate – permeability relationship to verify Darcy’s fluid flow law. The effect of model size scaling on sanding was also investigated using 4m height, 2m diameter model. The parallel bond model successfully calibrated the sample’s strength of 4.4MPa, showing a sharp peak strength before strain-softening, similar to the behavior of real cemented sandstones. There seems to be an exponential increasing relationship for the bigger model, but a curvilinear shape for the smaller model. The presence of the Darcy flow induced tensile forces and increased the number of broken bonds. For the parametric studies, flow rate has a linear relationship with permeability at constant pressure head. The higher the fluid flow pressure, the higher the number of broken bonds/sanding. The DEM PFC3D is a promising tool to studying the micromechanical behavior of cemented sandstones.

Keywords: discrete element method, fluid flow, parametric study, sand production/bonds failure

Procedia PDF Downloads 294

Authors: Yuvraj Singh Dangi, Brajesh Kumar Tiwari, Ashok Kumar Jain, Kamta Prasad Namdeo

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The potential use of liposomes as drug carriers by i.v. injection is limited by their low stability in blood stream. Firstly, phospholipid exchange and transfer to lipoproteins, mainly HDL destabilizes and disintegrates liposomes with subsequent loss of content. To avoid the pain associated with injection and to obtain better patient compliance studies concerning various dosage forms, have been developed. Conventional liposomes (unilamellar and multilamellar) have certain drawbacks like low entrapment efficiency, stability and release of drug after single breach in external membrane, have led to the new type of liposomal systems. The challenge has been successfully met in the form of Double Liposomes (DL). DL is a recently developed type of liposome, consisting of smaller liposomes enveloped in lipid bilayers. The outer lipid layer of DL can protect inner liposomes against various enzymes, therefore DL was thought to be more effective than ordinary liposomes. This concept was also supported by in vitro release characteristics i.e. DL formation inhibited the release of drugs encapsulated in inner liposomes. DL consists of several small liposomes encapsulated in large liposomes, i.e., multivesicular vesicles (MVV), therefore, DL should be discriminated from ordinary classification of multilamellar vesicles (MLV), large unilamellar vesicles (LUV), small unilamellar vesicles (SUV). However, for these liposomes, the volume of inner phase is small and loading volume of water-soluble drugs is low. In the present study, the potential of phosphatidylethanolamine (PE) lipid anchored double liposomes (DL) to incorporate two drugs in a single system is exploited as a tool to augment the H. pylori eradication rate. Preparation of DL involves two steps, first formation of primary (inner) liposomes by thin film hydration method containing one drug, then addition of suspension of inner liposomes on thin film of lipid containing the other drug. The success of formation of DL was characterized by optical and transmission electron microscopy. Quantitation of DL-bacterial interaction was evaluated in terms of percent growth inhibition (%GI) on reference strain of H. pylori ATCC 26695. To confirm specific binding efficacy of DL to H. pylori PE surface receptor we performed an agglutination assay. Agglutination in DL treated H. pylori suspension suggested selectivity of DL towards the PE surface receptor of H. pylori. Monotherapy is generally not recommended for treatment of a H. pylori infection due to the danger of development of resistance and unacceptably low eradication rates. Therefore, combination therapy with amoxicillin trihydrate (AMOX) as anti-H. pylori agent and ranitidine bismuth citrate (RBC) as antisecretory agent were selected for the study with an expectation that this dual-drug delivery approach will exert acceptable anti-H. pylori activity.

Keywords: Helicobacter pylorI, amoxicillin trihydrate, Ranitidine Bismuth citrate, phosphatidylethanolamine, multi vesicular systems

Procedia PDF Downloads 179

Authors: Oleg Reva, Ilya Korotetskiy, Marina Lankina, Murat Kulmanov, Aleksandr Ilin

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Molecular docking approaches are widely used for design of new antibiotics and modeling of antibacterial activities of numerous ligands which bind specifically to active centers of indispensable enzymes and/or key signaling proteins of pathogens. Widespread drug resistance among pathogenic microorganisms calls for development of new antibiotics specifically targeting important metabolic and information pathways. A generally recognized problem is that almost all molecular targets have been identified already and it is getting more and more difficult to design innovative antibacterial compounds to combat the drug resistance. A promising way to overcome the drug resistance problem is an induction of reversion of drug resistance by supplementary medicines to improve the efficacy of the conventional antibiotics. In contrast to well established computer-based drug design, modeling of drug resistance reversion still is in its infancy. In this work, we proposed an approach to identification of compensatory genetic variants reducing the fitness cost associated with the acquisition of drug resistance by pathogenic bacteria. The approach was based on an analysis of the population genetic of Mycobacterium tuberculosis and on results of experimental modeling of the drug resistance reversion induced by a new anti-tuberculosis drug FS-1. The latter drug is an iodine-containing nanomolecular complex that passed clinical trials and was admitted as a new medicine against MDR-TB in Kazakhstan. Isolates of M. tuberculosis obtained on different stages of the clinical trials and also from laboratory animals infected with MDR-TB strain were characterized by antibiotic resistance, and their genomes were sequenced by the paired-end Illumina HiSeq 2000 technology. A steady increase in sensitivity to conventional anti-tuberculosis antibiotics in series of isolated treated with FS-1 was registered despite the fact that the canonical drug resistance mutations identified in the genomes of these isolates remained intact. It was hypothesized that the drug resistance phenotype in M. tuberculosis requires an adjustment of activities of many genes to compensate the fitness cost of the drug resistance mutations. FS-1 cased an aggravation of the fitness cost and removal of the drug-resistant variants of M. tuberculosis from the population. This process caused a significant increase in genetic heterogeneity of the Mtb population that was not observed in the positive and negative controls (infected laboratory animals left untreated and treated solely with the antibiotics). A large-scale search for linkage disequilibrium associations between the drug resistance mutations and genetic variants in other genomic loci allowed identification of target proteins, which could be influenced by supplementary drugs to increase the fitness cost of the drug resistance and deprive the drug-resistant bacterial variants of their competitiveness in the population. The approach will be used to improve the efficacy of FS-1 and also for computer-based design of new drugs to combat drug-resistant infections.

Keywords: complete genome sequencing, computational modeling, drug resistance reversion, Mycobacterium tuberculosis

Procedia PDF Downloads 239

Authors: De-Shin Liu, Po-Chun Wen, Zhen-Wei Zhuang, Hsueh-Chih Liu, Pei-Chen Huang

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Wafer probing tests play an important role in semiconductor manufacturing procedures in accordance with the yield and reliability requirement of the wafer after the backend-of-the-line process. Accordingly, the stable physical and electrical contact between the probe and the tested wafer during wafer probing is regarded as an essential issue in identifying the known good die. The probe card can be integrated with multiple probe needles, which are classified as vertical, cantilever and micro-electro-mechanical systems type probe selections. Among all potential probe types, the vertical probe has several advantages as compared with other probe types, including maintainability, high probe density and feasibility for high-speed wafer testing. In the present study, the mechanical response of the wafer probing test with the vertical cobra probe on 720 μm thick silicon (Si) substrate with a 1.4 μm thick aluminum (Al) pad is investigated by the experiment and transient dynamic simulation approach. Because the deformation mechanism of the vertical cobra probe is determined by both bending and buckling mechanisms, the stable correlation between contact forces and overdrive (OD) length must be carefully verified. Moreover, the decent OD length with corresponding contact force contributed to piercing the native oxide layer of the Al pad and preventing the probing test-induced damage on the interconnect system. Accordingly, the scratch depth of the Al pad under various OD lengths is estimated by the atomic force microscope (AFM) and simulation work. In the wafer probing test configuration, the contact phenomenon between the probe needle and the tested object introduced large deformation and twisting of mesh gridding, causing the subsequent numerical divergence issue. For this reason, the arbitrary Lagrangian-Eulerian method is utilized in the present simulation work to conquer the aforementioned issue. The analytic results revealed a slight difference when the OD is considered as 40 μm, and the simulated is almost identical to the measured scratch depths of the Al pad under higher OD lengths up to 70 μm. This phenomenon can be attributed to the unstable contact of the probe at low OD length with the scratch depth below 30% of Al pad thickness, and the contact status will be being stable when the scratch depth over 30% of pad thickness. The splash of the Al pad is observed by the AFM, and the splashed Al debris accumulates on a specific side; this phenomenon is successfully simulated in the transient dynamic simulation. Thus, the preferred testing OD lengths are found as 45 μm to 70 μm, and the corresponding scratch depths on the Al pad are represented as 31.4% and 47.1% of Al pad thickness, respectively. The investigation approach demonstrated in this study contributed to analyzing the mechanical response of wafer probing test configuration under large strain conditions and assessed the geometric designs and material selections of probe needles to meet the requirement of high resolution and high-speed wafer-level probing test for thinned wafer application.

Keywords: wafer probing test, vertical probe, probe mark, mechanical response, FEA simulation

Procedia PDF Downloads 27

Authors: Hadi S. Hosseini, Larry A. Taber

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In the embryo, heart is initially a simple tubular structure that undergoes complex morphological changes as it transforms into a four-chambered pump. This work focuses on mechanisms that create heart tube (HT). The early embryo is composed of three relatively flat primary germ layers called endoderm, mesoderm, and ectoderm. Precardiac cells located within bilateral regions of the mesoderm called heart fields (HFs) fold and fuse along the embryonic midline to create the HT. The right and left halves of this plate fold symmetrically to bring their upper edges into contact along the midline, where they fuse. In a region near the fusion line, these layers then separate to generate the primitive HT and foregut, which then extend vertically. The anterior intestinal portal (AIP) is the opening at the caudal end of the foregut, which descends as the HT lengthens. The biomechanical mechanisms that drive this folding are poorly understood. Our central hypothesis is that folding is caused by differences in growth between the endoderm and mesoderm while subsequent extension is driven by contraction along the AIP. The feasibility of this hypothesis is examined using experiments with chick embryos and finite-element modeling (FEM). Fertilized white Leghorn chicken eggs were incubated for approximately 22-33 hours until appropriate Hamburger and Hamilton stage (HH5 to HH9) was reached. To inhibit contraction, embryos were cultured in media containing blebbistatin (myosin II inhibitor) for 18h. Three-dimensional models were created using ABAQUS (D. S. Simulia). The initial geometry consists of a flat plate including two layers representing the mesoderm and endoderm. Tissue was considered as a nonlinear elastic material with growth and contraction (negative growth) simulated using a theory, in which the total deformation gradient is given by F=F^*.G, where G is growth tensor and F* is the elastic deformation gradient tensor. In embryos exposed to blebbistatin, initial folding and AIP descension occurred normally. However, after HFs partially fused to create the upper part of the HT, fusion, and AIP descension stopped, and the HT failed to grow longer. These results suggest that cytoskeletal contraction is required only for the later stages of HT formation. In the model, a larger biaxial growth rate in the mesoderm compared to the endoderm causes the bilayered plate to bend ventrally, as the upper edge moves toward the midline, where it 'fuses' with the other half . This folding creates the upper section of the HT, as well as the foregut pocket bordered by the AIP. After this phase completes by stage HH7, contraction along the arch-shaped AIP pulls the lower edge of the plate downward, stretching the two layers. Results given by model are in reasonable agreement with experimental data for the shape of HT, as well as patterns of stress and strain. In conclusion, results of our study support our hypothesis for the creation of the heart tube.

Keywords: heart tube formation, FEM, chick embryo, biomechanics

Procedia PDF Downloads 278

Authors: Jone Ibarruri, Mikel Manso, Marta Cebrián

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A high amount of food is lost or discarded in the World every year. In addition, in the last decades, an increasing demand of new alternative and sustainable sources of proteins and other valuable compounds is being observed in the food and feeding sectors and, therefore, the use of food by-products as nutrients for these purposes sounds very interesting from the environmental and economical point of view. However, the direct use of discarded fruit and vegetables that present, in general, a low protein content is not interesting as feeding ingredient except if they are used as a source of fiber for ruminants. Especially in the case of aquaculture, several alternatives to the use of fish meal and other vegetable protein sources have been extensively explored due to the scarcity of fish stocks and the unsustainability of fishing for these purposes. Fish mortality is also of great concern in this sector as this problem highly reduces their economic feasibility. So, the development of new functional and natural ingredients that could reduce the need for vaccination is also of great interest. In this work, several fermentation tests were developed at lab scale using a selected mixture of fruit and vegetable discards from a wholesale market located in the Basque Country to increase their protein content and also to produce some bioactive extracts that could be used as additives in aquaculture. Fruit and vegetable mixtures (60/40 ww) were centrifugated for humidity reduction and crushed to 2-5 mm particle size. Samples were inoculated with a selected Rhizopus oryzae strain and fermented for 7 days in controlled conditions (humidity between 65 and 75% and 28ºC) in Petri plates (120 mm) by triplicate. Obtained results indicated that the final fermented product presented a twofold protein content (from 13 to 28% d.w). Fermented product was further processed to determine their possible functionality as a feed additive. Extraction tests were carried out to obtain an ethanolic extract (60:40 ethanol: water, v.v) and remaining biomass that also could present applications in food or feed sectors. The extract presented a polyphenol content of about 27 mg GAE/gr d.w with antioxidant activity of 8.4 mg TEAC/g d.w. Remining biomass is mainly composed of fiber (51%), protein (24%) and fat (10%). Extracts also presented antibacterial activity according to the results obtained in Agar Diffusion and to the Minimum Inhibitory Concentration (MIC) tests determined against several food and fish pathogen strains. In vitro, digestibility was also assessed to obtain preliminary information about the expected effect of extraction procedure on fermented product digestibility. First results indicated that remaining biomass after extraction doesn´t seem to improve digestibility in comparison to the initial fermented product. These preliminary results show that fermented fruit and vegetables can be a useful source of functional ingredients for aquaculture applications and a substitute of other protein sources in the feeding sector. Further validation will be also carried out through “in vivo” tests with trout and bass.

Keywords: fungal solid state fermentation, protein increase, functional extracts, feed ingredients

Procedia PDF Downloads 40

Authors: Alvarez M., Chapela M. J., Balboa E., Rubianes D., Sinde E., Fernandez de Ana C., Rodríguez-Blanco A.

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The gut microbiota plays an important role as gut inflammation could contribute to colorectal cancer development; however, this role is still not fully understood, and tools able to prevent this progression are yet to be developed. The main objective of this study was to monitor the effects of a mushroom extracts formulation in gut microbial community composition of an Azoxymethane (AOM)/Dextran sodium sulfate (DSS) mice model of inflammation-linked carcinogenesis. For the in vivo study, 41 adult male mice of the C57BL / 6 strain were obtained. 36 of them have been induced in a state of colon carcinogenesis by a single intraperitoneal administration of AOM at a dose of 12.5 mg/kg; the control group animals received instead of the same volume of 0.9% saline. DSS is an extremely toxic polysaccharide sulfate that causes chronic inflammation of the colon mucosa, favoring the appearance of severe colitis and the production of tumors induced by AOM. Induction by AOM/DSS is an interesting platform for chemopreventive intervention studies. This time the model was used to monitor gut microbiota changes as a result of supplementation with a specific mushroom extracts formulation previously shown to have prebiotic activity. The animals have been divided into three groups: (i) Cancer + mushroom extracts formulation experimental group: to which the MicoDigest2.0 mushroom extracts formulation developed by Hifas da Terra S.L has been administered dissolved in drinking water at an estimated concentration of 100 mg / ml. (ii) Control group of animals with Cancer: to which normal water has been administered without any type of treatment. (iii) Control group of healthy animals: these are the animals that have not been induced cancer or have not received any treatment in drinking water. This treatment has been maintained for a period of 3 months, after which the animals were sacrificed to obtain tissues that were subsequently analyzed to verify the effects of the mushroom extract formulation. A microbiological analysis has been carried out to compare the microbial communities present in the intestines of the mice belonging to each of the study groups. For this, the methodology of massive sequencing by molecular analysis of the 16S gene has been used (Ion Torrent technology). Initially, DNA extraction and metagenomics libraries were prepared using the 16S Metagenomics kit, always following the manufacturer's instructions. This kit amplifies 7 of the 9 hypervariable regions of the 16S gene that will then be sequenced. Finally, the data obtained will be compared with a database that makes it possible to determine the degree of similarity of the sequences obtained with a wide range of bacterial genomes. Results obtained showed that, similarly to certain natural compounds preventing colorectal tumorigenesis, a mushroom formulation enriched the Firmicutes and Proteobacteria phyla and depleted Bacteroidetes. Therefore, it was demonstrated that the consumption of the mushroom extracts’ formulation developed could promote the recovery of the microbial balance that is disrupted in the mice model of carcinogenesis. More preclinical and clinical studies are needed to validate this promising approach.

Keywords: carcinogenesis, microbiota, mushroom extracts, inflammation

Procedia PDF Downloads 129

Authors: Sergey A. Lurie, Yury O. Solyaev, Dmitry B. Volkov-Bogorodsky, Alexander V. Volkov

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The media with voids is considered and the method of the analytical estimation of the effective mechanical properties in the theory of elastic materials with voids is proposed. The variational model of the porous media is discussed, which is based on the model of the media with fields of conserved dislocations. It is shown that this model is fully consistent with the known model of the linear elastic materials with voids. In the present work, the generalized model of the porous media is proposed in which the specific surface properties are associated with the field of defects-pores in the volume of the deformed body. Unlike typical surface elasticity model, the strain energy density of the considered model includes the special part of the surface energy with the quadratic form of the free distortion tensor. In the result, the non-classical boundary conditions take modified form of the balance equations of volume and surface stresses. The analytical approach is proposed in the present work which allows to receive the simple enough engineering estimations for effective characteristics of the media with free dilatation. In particular, the effective flexural modulus and Poisson's ratio are determined for the problem of a beam pure bending. Here, the known voids elasticity solution was expanded on the generalized model with the surface effects. Received results allow us to compare the deformed state of the porous beam with the equivalent classic beam to introduce effective bending rigidity. Obtained analytical expressions for the effective properties depend on the thickness of the beam as a parameter. It is shown that the flexural modulus of the porous beam is decreased with an increasing of its thickness and the effective Poisson's ratio of the porous beams can take negative values for the certain values of the model parameters. On the other hand, the effective shear modulus is constant under variation of all values of the non-classical model parameters. Solutions received for a beam pure bending and the hydrostatic loading of the porous media are compared. It is shown that an analytical estimation for the bulk modulus of the porous material under hydrostatic compression gives an asymptotic value for the effective bulk modulus of the porous beam in the case of beam thickness increasing. Additionally, it is shown that the scale effects appear due to the surface properties of the porous media. Obtained results allow us to offer the procedure of an experimental identification of the non-classical parameters in the theory of the linear elastic materials with voids based on the bending tests for samples with different thickness. Finally, the problem of implementation of the Saint-Venant hypothesis for the transverse stresses in the porous beam are discussed. These stresses are different from zero in the solution of the voids elasticity theory, but satisfy the integral equilibrium equations. In this work, the exact value of the introduced surface parameter was found, which provides the vanishing of the transverse stresses on the free surfaces of a beam.

Keywords: effective properties, scale effects, surface defects, voids elasticity

Procedia PDF Downloads 379

Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

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Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

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Authors: Adil Murtaza, Muhammad Tahir Khan, Awais Ghani, Chao Zhou, Sen Yang, Xiaoping Song

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The morphotropic phase boundary (MPB); a boundary between two different crystallographic symmetries in the composition–temperature phase diagram has been widely studied in ferroelectrics and recently has drawn interest in ferromagnets for obtaining enhanced large field-induced strain. At MPB, the system gets a compressed free energy state, which allows the polarization to freely rotate and hence results in a high magnetoelastic response (e.g., high magnetization, low coercivity, and large magnetostriction). Based on the same mechanism, we designed MPB in a ferromagnetic Tb₁₋ₓNdₓCo₂ system. The temperature-dependent magnetization curves showed spin reorientation (SR); which can be explained by a two-sublattice model. Contrary to previously reported MPB involved ferromagnetic systems, the MPB composition of Tb₀.₃₅Nd₀.₆₅Co₂ exhibits a low saturation magnetization (MS), indicating a compensation of the Tb and Nd magnetic moments at MPB. The coercive field (HC) under a low magnetic field and first anisotropy constant (K₁) shows a minimum value at MPB composition of x=0.65. A detailed spin configuration diagram is provided for the Tb₁₋ₓNdₓCo₂ around the composition for the anisotropy compensation; this can guide the development of novel magnetostrictive materials. The anisotropic magnetostriction (λS) first decreased until x=0.8 and then continuously increased in the negative direction with further increase of Nd concentration. In addition, the large ratio between magnetostriction and the absolute values of the first anisotropy constant (λS/K₁) appears at MPB, indicating that Tb₀.₃₅Nd₀.₆₅Co₂ has good magnetostrictive properties. Present work shows an anomalous type of MPB in ferromagnetic materials, revealing that MPB can also lead to a weakening of magnetoelastic behavior as shown in the ferromagnetic Tb₁₋ₓNdₓCo₂ system. Our work shows the universal presence of MPB in ferromagnetic materials and suggests the differences between different ferromagnetic MPB systems that are important for substantial improvement of magnetic and magnetostrictive properties. Based on the results of this study, similar MPB effects might be achieved in other ferroic systems that can be used for technological applications. The finding of magnetic MPB in the ferromagnetic system leads to some important significances. First, it provides a better understanding of the fundamental concept of spin reorientation transitions (SRT) like ferro-ferro transitions are not only reorientation of magnetization but also crystal symmetry change upon magnetic ordering. Second, the flattened free energy corresponding to a low energy barrier for magnetization rotation and enhanced magnetoelastic response near MPB. Third, to attain large magnetostriction with MPB approach two terminal compounds have different easy magnetization directions below Curie temperature Tc in order to accomplish the weakening of magnetization anisotropy at MPB (as in ferroelectrics), thus easing the magnetic domain switching and the lattice distortion difference between two terminal compounds should be large enough, e.g., lattice distortion of R symmetry ˃˃ lattice distortion of T symmetry). So that the MPB composition agrees to a nearly isotropic state along with large ‘net’ lattice distortion, which is revealed in a higher value of magnetostriction.

Keywords: magnetization, magnetostriction, morphotropic phase boundary (MPB), phase transition

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Authors: Hady Hamidyan

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As the global population continues to grow, the demand for clean water and effective wastewater treatment becomes increasingly critical. By 2030, global water demand is projected to exceed supply by 40%, driven by population growth, increased water usage, and climate change. Currently, about 4.2 billion people lack access to safely managed sanitation services. The wastewater treatment sector faces numerous challenges, including the need for energy-efficient solutions, cost-effectiveness, ease of use, and low maintenance requirements. This abstract presents a groundbreaking wastewater treatment technology that addresses these challenges by offering an energy-saving approach, wastewater recovery capabilities, and a ready-made, affordable, and user-friendly package with minimal maintenance costs. The unique design of this ready-made package made it possible to eliminate the need for pumps, filters, airlift, and other common equipment. Consequently, it enables sustainable wastewater treatment management with exceptionally low energy and cost requirements, minimizing investment and maintenance expenses. The operation of these packages is based on continuous aeration, which involves injecting oxygen gas or air into the aeration chamber through a tubular diffuser with very small openings. This process supplies the necessary oxygen for aerobic bacteria. The recovered water, which amounts to almost 95% of the input, can be treated to meet specific quality standards, allowing safe reuse for irrigation, industrial processes, or even potable purposes. This not only reduces the strain on freshwater resources but also provides economic benefits by offsetting the costs associated with freshwater acquisition and wastewater discharge. The ready-made, affordable, and user-friendly nature of this technology makes it accessible to a wide range of users, including small communities, industries, and decentralized wastewater treatment systems. The system incorporates user-friendly interfaces, simplified operational procedures, and integrated automation, facilitating easy implementation and operation. Additionally, the use of durable materials, efficient equipment, and advanced monitoring systems significantly reduces maintenance requirements, resulting in low overall life-cycle costs and alleviating the burden on operators and maintenance personnel. In conclusion, the presented wastewater treatment technology offers a comprehensive solution to the challenges faced by the industry. Its energy-saving approach, combined with wastewater recovery capabilities, ensures sustainable resource management and enhances environmental stewardship. This affordable, ready-made, and low-maintenance package promotes broad adoption across various sectors and communities, contributing to a more sustainable future for water and wastewater management.

Keywords: wastewater treatment, energy saving, wastewater recovery, affordable package, low maintenance costs, sustainable resource management, environmental stewardship

Procedia PDF Downloads 51

Authors: Zanele D. Ngwenya, Mustapha Mohammed, Felix D. Dakora

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Legumes are a major source of biological nitrogen, and therefore play a crucial role in maintaining soil productivity in smallholder agriculture in southern Africa. Through their ability to fix atmospheric nitrogen in root nodules, legumes are a better option for sustainable nitrogen supply in cropping systems than chemical fertilisers. For decades, farmers have been highly receptive to the use of rhizobial inoculants as a source of nitrogen due mainly to the availability of elite rhizobial strains at a much lower compared to chemical fertilisers. To improve the efficiency of the legume-rhizobia symbiosis in African soils would require the use of highly effective rhizobia capable of nodulating a wide range of host plants. This study assessed the morphogenetic diversity, photosynthetic functioning and relative symbiotic effectiveness (RSE) of groundnut, jack bean and soybean microsymbionts in Eswatini soils as a first step to identifying superior isolates for inoculant production. According to the manufacturer's instructions, rhizobial isolates were cultured in yeast-mannitol (YM) broth until the late log phase and the bacterial genomic DNA was extracted using GenElute bacterial genomic DNA kit. The extracted DNA was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and a dendrogram constructed from the band patterns to assess rhizobial diversity. To assess the N2-fixing efficiency of the authenticated rhizobia, photosynthetic rates (A), stomatal conductance (gs), and transpiration rates (E) were measured at flowering for plants inoculated with the test isolates. The plants were then harvested for nodulation assessment and measurement of plant growth as shoot biomass. The results of ERIC-PCR fingerprinting revealed the presence of high genetic diversity among the microsymbionts nodulating each of the three test legumes, with many of them showing less than 70% ERIC-PCR relatedness. The dendrogram generated from ERIC-PCR profiles grouped the groundnut isolates into 5 major clusters, while the jack bean and soybean isolates were grouped into 6 and 7 major clusters, respectively. Furthermore, the isolates also elicited variable nodule number per plant, nodule dry matter, shoot biomass and photosynthetic rates in their respective host plants under glasshouse conditions. Of the groundnut isolates tested, 38% recorded high relative symbiotic effectiveness (RSE >80), while 55% of the jack bean isolates and 93% of the soybean isolates recorded high RSE (>80) compared to the commercial Bradyrhizobium strains. About 13%, 27% and 83% of the top N₂-fixing groundnut, jack bean and soybean isolates, respectively, elicited much higher relative symbiotic efficiency (RSE) than the commercial strain, suggesting their potential for use in inoculant production after field testing. There was a tendency for both low and high N₂-fixing isolates to group together in the dendrogram from ERIC-PCR profiles, which suggests that RSE can differ significantly among closely related microsymbionts.

Keywords: genetic diversity, relative symbiotic effectiveness, inoculant, N₂-fixing

Procedia PDF Downloads 185

Authors: Sumeet Rai, Anuj Tyagi, B. T. Naveen Kumar, Shubhkaramjeet Kaur, Niraj K. Singh

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Since their discovery, indiscriminate use of antibiotics in human, veterinary and aquaculture systems has resulted in global emergence/spread of multidrug-resistant bacterial pathogens. Thus, the need for alternative approaches to control bacterial infections has become utmost important. High selectivity/specificity of bacteriophages (phages) permits the targeting of specific bacteria without affecting the desirable flora. In this study, a lytic phage (Ahp1) specific to Aeromonas hydrophila subsp. hydrophila was isolated from finfish aquaculture pond. The host range of Ahp1 range was tested against 10 isolates of A. hydrophila, 7 isolates of A. veronii, 25 Vibrio cholerae isolates, 4 V. parahaemolyticus isolates and one isolate each of V. harveyi and Salmonella enterica collected previously. Except the host A. hydrophila subsp. hydrophila strain, no lytic activity against any other bacterial was detected. During the adsorption rate and one-step growth curve analysis, 69.7% of phage particles were able to get adsorbed on host cell followed by the release of 93 ± 6 phage progenies per host cell after a latent period of ~30 min. Phage nucleic acid was extracted by column purification methods. After determining the nature of phage nucleic acid as dsDNA, phage genome was subjected to next-generation sequencing by generating paired-end (PE, 2 x 300bp) reads on Illumina MiSeq system. De novo assembly of sequencing reads generated circular phage genome of 42,439 bp with G+C content of 58.95%. During open read frame (ORF) prediction and annotation, 22 ORFs (out of 49 total predicted ORFs) were functionally annotated and rest encoded for hypothetical proteins. Proteins involved in major functions such as phage structure formation and packaging, DNA replication and repair, DNA transcription and host cell lysis were encoded by the phage genome. The complete genome sequence of Ahp1 along with gene annotation was submitted to NCBI GenBank (accession number MF683623). Stability of Ahp1 preparations at storage temperatures of 4 °C, 30 °C, and 40 °C was studied over a period of 9 months. At 40 °C storage, phage counts declined by 4 log units within one month; with a total loss of viability after 2 months. At 30 °C temperature, phage preparation was stable for < 5 months. On the other hand, phage counts decreased by only 2 log units over a period of 9 during storage at 4 °C. As some of the phages have also been reported as glycerol sensitive, the stability of Ahp1 preparations in (0%, 15%, 30% and 45%) glycerol stocks were also studied during storage at -80 °C over a period of 9 months. The phage counts decreased only by 2 log units during storage, and no significant difference in phage counts was observed at different concentrations of glycerol. The Ahp1 phage discovered in our study had a very narrow host range and it may be useful for phage typing applications. Moreover, the endolysin and holin genes in Ahp1 genome could be ideal candidates for recombinant cloning and expression of antimicrobial proteins.

Keywords: Aeromonas hydrophila, endolysin, phage, narrow host range

Procedia PDF Downloads 144

Authors: Muhammad Tariq, Muhammad Waseem, Muhammad Hidayat Rasool

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Isolation and characterization of chromium tolerant Staphylococcus aureus from industrial wastewater and their potential use to bioremediate environmental chromium. Objectives: Chromium with its great economic importance in industrial use is major metal pollutant of the environment. Chromium are used in different industries for various applications such as textile, dyeing and pigmentation, wood preservation, manufacturing pulp and paper, chrome plating, steel and tanning. The release of untreated chromium in industrial effluents causes serious threat to environment and human health, therefore, the current study designed to isolate chromium tolerant Staphylococcus aureus for removal of chromium prior to their final discharge into the environment due to its cost effective and beneficial advantage over physical and chemical methods. Methods: Wastewater samples were collected from discharge point of different industries. Heavy metal analysis by atomic absorption spectrophotometer and microbiological analysis such as total viable count, total coliform, fecal coliform and Escherichia coli were conducted. Staphylococcus aureus was identified through gram’s staining, biomeriux vitek 2 microbial identification system and 16S rRNA gene amplification by polymerase chain reaction. Optimum growth conditions with respect to temperature, pH, salt concentrations and effect of chromium on the growth of bacteria, resistance to other heavy metal ions, minimum inhibitory concentration and chromium uptake ability of Staphylococcus aureus strain K1 was determined by spectrophotometer. Antibiotic sensitivity pattern was also determined by disc diffusion method. Furthermore, chromium uptake ability was confirmed by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope equipped with Oxford Energy Dipersive X-ray (EDX) micro analysis system. Results: The results presented that optimum temperature was 35ᵒC, pH was 8.0 and salt concentration was 0.5% for growth of Staphylococcus aureus K1. The maximum uptake ability of chromium by bacteria was 20mM than other heavy metal ions. The antibiotic sensitivity pattern revealed that Staphylococcus aureus was vancomycin and methicillin sensitive. Non hemolytic activity on blood agar and negative coagulase reaction showed that it was non-pathogenic. Furthermore, the growth of bacteria decreases in the presence of chromium and maximum chromium uptake by bacteria observed at optimum growth conditions. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and Energy dispersive X-ray (EDX) analysis confirmed the presence of chromium uptake by Staphylococcus aureus K1. Conclusion: The study revealed that Staphylococcus aureus K1 have the potential to bio-remediate chromium toxicity from wastewater. Gradually, this biological treatment becomes more important due to its advantage over physical and chemical methods to protect environment and human health.

Keywords: wastewater, staphylococcus, chromium, bioremediation

Procedia PDF Downloads 143