Search results for: brine shrimp assay (BSA)
1274 YHV-Responsive Gene Expression under the Influence of PmRelish Regulation
Authors: Suwattana Visetnan, Premruethai Supungul, Sureerat Tang, Ikuo Hirono, Anchalee Tassanakajon, Vichien Rimphanitchayakit
Abstract:
In animals, infection by Gram-negative bacteria and certain viruses activates the Imd signaling pathway wherein the a NF-κB transcription factor, Relish, is a key regulatory protein for the synthesis of antimicrobial proteins. Infection by yellow head virus (YHV) activates the Imd pathway. To investigate the expression of genes involved in YHV infection and under the influence of PmRelish regulation, RNA interference and suppression subtractive hybridization (SSH) are employed. The genes in forward library expressed in shrimp after YHV infection and under the activity of PmRelish were obtained by subtracting the cDNAs from YHV-infected and PmRelish-knockdown shrimp with cDNAs from YHV-infected shrimp. Opposite subtraction gave a reverse library whereby an alternative set of genes under YHV infection and no PmRelish expression was obtained. Sequencing of 252 and 99 cDNA clones from the respective forward and reverse libraries were done and annotated through blast search against the GenBank sequences. Genes involved in defense and homeostasis were abundant in both libraries, 31% and 23% in the forward and reverse libraries, respectively. They were predominantly antimicrobial proteins, proteinases and proteinase inhibitors. The expression of antimicrobial protein genes, ALFPm3, crustinPm1, penaeidin3 and penaeidin5 were tested under PmRelish silencing and Gram-negative bacterium V. harveyi infection. Together with the results previously reported, the expression of penaeidin5 and also penaeidin3 but not ALFPm3 and crustinPm1 were under the regulation of PmRelish in the Imd pathway.Keywords: relish, yellow head virus, penaeus monodon, antimicrobial proteins
Procedia PDF Downloads 2121273 Prevalence of Enterocytozoon hepatopenaei in Shrimp Cultured in Inland Saline Water
Authors: Naveen Kumar B. T., Anuj Tyagi, Prabjeet Singh, Shanthanagouda A. H., Sumeet Rai
Abstract:
Inland saline water resources are gaining the importance in expanding the aquaculture activities to mitigate the nutritional and food security issues of the world. For profitable and sustainable aquaculture practices, scientific farming, biosecurity measure, and best fish health management should be the integral part of developmental activities. Keeping in line with global awareness and trends, the Indian government has taken an innovative step to conduct disease surveillance and awareness programme for aquatic disease through network project. This ‘National Surveillance Programme for Aquatic Animal Diseases (NSPAAD)’ is being implemented in collaboration of national institutes and state agriculture universities with funding support from National Fisheries Development Board (NFDB), Govt. of India. Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, an NSPAAD collaborator, has been actively engaged in disease surveillance in the Indian state of Punjab. Shrimp farming in inland saline areas of Punjab is expanding at a tremendous pace under the guidance of GADVASU along with the support of State Fisheries Department. Under this national disease surveillance programme, we reported Enterocytozoon hepatopenaei (EHP) infection in the Litopenaeus vannamei cultured in the inland saline waters. Polymerase chain reaction (PCR) based diagnosis was carried out using the OIE (World Organisation for Animal Health) protocol. It was observed that out of 20 shrimp farms, two farms were 1st step PCR positive and two more farms were nested PCR positive. All the EHP positive ponds had shown the white faeces along with mortalities at very low rate. Therefore, implementation of biosecurity and continuous surveillance and monitoring program for finfish and shellfish aquaculture are in need of the hour to prevent and control the large-scale disease outbreaks and subsequent economic losses.Keywords: disease, EHP, inland saline water, shrimp culture
Procedia PDF Downloads 2621272 Effect of Oil Viscosity and Brine Salinity/Viscosity on Water/Oil Relative Permeability and Residual Saturations
Authors: Sami Aboujafar
Abstract:
Oil recovery in petroleum reservoirs is greatly affected by fluid-rock and fluid-fluid interactions. These interactions directly control rock wettability, capillary pressure and relative permeability curves. Laboratory core-floods and centrifuge experiments were conducted on sandstone and carbonate cores to study the effect of low and high brine salinity and viscosity and oil viscosity on residual saturations and relative permeability. Drainage and imbibition relative permeability in two phase system were measured, refined lab oils with different viscosities, heavy and light, and several brine salinities were used. Sensitivity analysis with different values for the salinity and viscosity of the fluids,, oil and water, were done to investigate the effect of these properties on water/oil relative permeability, residual oil saturation and oil recovery. Experiments were conducted on core material from viscous/heavy and light oil fields. History matching core flood simulator was used to study how the relative permeability curves and end point saturations were affected by different fluid properties using several correlations. Results were compared with field data and literature data. The results indicate that there is a correlation between the oil viscosity and/or brine salinity and residual oil saturation and water relative permeability end point. Increasing oil viscosity reduces the Krw@Sor and increases Sor. The remaining oil saturation from laboratory measurements might be too high due to experimental procedures, capillary end effect and early termination of the experiment, especially when using heavy/viscous oil. Similarly the Krw@Sor may be too low. The effect of wettability on the observed results is also discussed. A consistent relationship has been drawn between the fluid parameters, water/oil relative permeability and residual saturations, and a descriptor may be derived to define different flow behaviors. The results of this work will have application to producing fields and the methodologies developed could have wider application to sandstone and carbonate reservoirs worldwide.Keywords: history matching core flood simulator, oil recovery, relative permeability, residual saturations
Procedia PDF Downloads 3371271 Optimization of Assay Parameters of L-Glutaminase from Bacillus cereus MTCC1305 Using Artificial Neural Network
Authors: P. Singh, R. M. Banik
Abstract:
Artificial neural network (ANN) was employed to optimize assay parameters viz., time, temperature, pH of reaction mixture, enzyme volume and substrate concentration of L-glutaminase from Bacillus cereus MTCC 1305. ANN model showed high value of coefficient of determination (0.9999), low value of root mean square error (0.6697) and low value of absolute average deviation. A multilayer perceptron neural network trained with an error back-propagation algorithm was incorporated for developing a predictive model and its topology was obtained as 5-3-1 after applying Levenberg Marquardt (LM) training algorithm. The predicted activity of L-glutaminase was obtained as 633.7349 U/l by considering optimum assay parameters, viz., pH of reaction mixture (7.5), reaction time (20 minutes), incubation temperature (35˚C), substrate concentration (40mM), and enzyme volume (0.5ml). The predicted data was verified by running experiment at simulated optimum assay condition and activity was obtained as 634.00 U/l. The application of ANN model for optimization of assay conditions improved the activity of L-glutaminase by 1.499 fold.Keywords: Bacillus cereus, L-glutaminase, assay parameters, artificial neural network
Procedia PDF Downloads 4291270 Cadmium Removal from Aqueous Solution Using Chitosan Beads Prepared from Shrimp Shell Extracted Chitosan
Authors: Bendjaballah Malek; Makhlouf Mohammed Rabeh; Boukerche Imane; Benhamza Mohammed El Hocine
Abstract:
In this study, chitosan was derived from Parapenaeus longirostris shrimp shells sourced from a local market in Annaba, eastern Algeria. The extraction process entailed four chemical stages: demineralization, deproteinization, decolorization, and deacetylation. The degree of deacetylation was calculated to be 80.86 %. The extracted chitosan was physically altered to synthesize chitosan beads and characterized via FTIR and XRD analysis. These beads were employed to eliminate cadmium ions from synthetic water. The batch adsorption process was optimized by analyzing the impact of contact time, pH, adsorbent dose, and temperature. The adsorption capacity of and Cd+2 on chitosan beads was found to be 6.83 mg/g and 7.94 mg/g, respectively. The kinetic adsorption of Cd+2 conformed to the pseudo-first-order model, while the isotherm study indicated that the Langmuir Isotherm model well described the adsorption of cadmium . A thermodynamic analysis demonstrated that the adsorption of Cd+2 on chitosan beads is spontaneous and exothermic.Keywords: Cd, chitosan, chitosanbeds, bioadsorbent
Procedia PDF Downloads 1011269 Performance Assessment of the Gold Coast Desalination Plant Offshore Multiport Brine Diffuser during ‘Hot Standby’ Operation
Authors: M. J. Baum, B. Gibbes, A. Grinham, S. Albert, D. Gale, P. Fisher
Abstract:
Alongside the rapid expansion of Seawater Reverse Osmosis technologies there is a concurrent increase in the production of hypersaline brine by-products. To minimize environmental impact, these by-products are commonly disposed into open-coastal environments via submerged diffuser systems as inclined dense jet outfalls. Despite the widespread implementation of this process, diffuser designs are typically based on small-scale laboratory experiments under idealistic quiescent conditions. Studies concerning diffuser performance in the field are limited. A set of experiments were conducted to assess the near field characteristics of brine disposal at the Gold Coast Desalination Plant offshore multiport diffuser. The aim of the field experiments was to determine the trajectory and dilution characteristics of the plume under various discharge configurations with production ranging 66 – 100% of plant operative capacity. The field monitoring system employed an unprecedented static array of temperature and electrical conductivity sensors in a three-dimensional grid surrounding a single diffuser port. Complimenting these measurements, Acoustic Doppler Current Profilers were also deployed to record current variability over the depth of the water column and wave characteristics. Recorded data suggested the open-coastal environment was highly active over the experimental duration with ambient velocities ranging 0.0 – 0.5 m∙s-1, with considerable variability over the depth of the water column observed. Variations in background electrical conductivity corresponding to salinity fluctuations of ± 1.7 g∙kg-1 were also observed. Increases in salinity were detected during plant operation and appeared to be most pronounced 10 – 30 m from the diffuser, consistent with trajectory predictions described by existing literature. Plume trajectories and respective dilutions extrapolated from salinity data are compared with empirical scaling arguments. Discharge properties were found to adequately correlate with modelling projections. Temporal and spatial variation of background processes and their subsequent influence upon discharge outcomes are discussed with a view to incorporating the influence of waves and ambient currents in the design of brine outfalls into the future.Keywords: brine disposal, desalination, field study, negatively buoyant discharge
Procedia PDF Downloads 2391268 Modeling Core Flooding Experiments for Co₂ Geological Storage Applications
Authors: Avinoam Rabinovich
Abstract:
CO₂ geological storage is a proven technology for reducing anthropogenic carbon emissions, which is paramount for achieving the ambitious net zero emissions goal. Core flooding experiments are an important step in any CO₂ storage project, allowing us to gain information on the flow of CO₂ and brine in the porous rock extracted from the reservoir. This information is important for understanding basic mechanisms related to CO₂ geological storage as well as for reservoir modeling, which is an integral part of a field project. In this work, a different method for constructing accurate models of CO₂-brine core flooding will be presented. Results for synthetic cases and real experiments will be shown and compared with numerical models to exhibit their predictive capabilities. Furthermore, the various mechanisms which impact the CO₂ distribution and trapping in the rock samples will be discussed, and examples from models and experiments will be provided. The new method entails solving an inverse problem to obtain a three-dimensional permeability distribution which, along with the relative permeability and capillary pressure functions, constitutes a model of the flow experiments. The model is more accurate when data from a number of experiments are combined to solve the inverse problem. This model can then be used to test various other injection flow rates and fluid fractions which have not been tested in experiments. The models can also be used to bridge the gap between small-scale capillary heterogeneity effects (sub-core and core scale) and large-scale (reservoir scale) effects, known as the upscaling problem.Keywords: CO₂ geological storage, residual trapping, capillary heterogeneity, core flooding, CO₂-brine flow
Procedia PDF Downloads 701267 A Phylogenetic Analysis and Effect of NO₃ Regime on the Level of N-3 Polyunsaturated Fatty Acids in Thalassiosira weissflogii Isolated from Caspian Sea
Authors: Ehsan Etesami, Mostafa Noroozi
Abstract:
Thalassiosira weissflogii with proper size and nutrition value specially PUFA n-3 has been widely used in bivalve shellfish larviculture and shrimp industries. This diatom was isolated from Caspian Sea and identified with morphology and molecular characters. T. weissflogii was cultivated in normal and nitrogen deficiency F2 medium during 18 to 30 days, in addition, the growth indices, total lipid, and EPA-DHA content were elucidated. The growth indices of the cells decreased during the stress experiments while the total lipid levels increased during prolonged culturing (30 days). The maximum level of C20:5 was calculated as 8.8 (%TFA) in normal condition during 30 days; however, the combination of N- deficiency condition with prolonged culturing led to the increase of the level of C22:6 from 3.5 to 12.63 (%TFA). The concept of N-deficiency along with prolonged culturing of Thalassiosira weissflogii can improve PUFA n-3 content in order to use in shellfish and shrimp industries.Keywords: DHA, Thalassiosira weissflogii, nitrogen deficiency, EPA, fatty acids, aquafeed
Procedia PDF Downloads 1411266 Enhancing Heavy Oil Recovery: Experimental Insights into Low Salinity Polymer in Sandstone Reservoirs
Authors: Intisar, Khalifa, Salim, Al Busaidi
Abstract:
Recently, the synergic combination of low salinity water flooding with polymer flooding has been a subject of paramount interest for the oil industry. Numerous studies have investigated the efficiency of enhanced oil recovery using low salinity polymer flooding (LSPF). However, there is no clear conclusion that can explain the incremental oil recovery, determine the main factors controlling the oil recovery process, and define the relative contribution of rock/fluids or fluid/fluid interactions to extra oil recovery. Therefore, this study aims to perform a systematic investigation of the interactions between oil, polymer, low salinity and sandstone rock surface from pore to core scale during LSPF. Partially hydrolyzed polyacrylamide (HPAM) polymer, Boise outcrop, a crude oil sample and reservoir cores from an Omani oil field, and brine at two different salinities were used in the study. Several experimental measurements including static bulk measurements of polymer solutions prepared with brines of high and low salinities, single phase displacement experiments, along with rheological, total organic carbon and ion chromatography measurements to analyze ion exchange reactions, polymer adsorption, and viscosity loss were used. In addition, two-phase experiments were performed to demonstrate the oil recovery efficiency of LSPF. The results revealed that the incremental oil recovery from LSPF was attributed to the combination of the reduction in the water-oil mobility ratio, an increase in the repulsion forces between crude oil/brine/rock interfaces and an increase in pH of the aqueous solution. In addition, lowering the salinity of the make-up brine resulted in a larger conformation (expansion) of the polymer molecules, which in turn resulted in less adsorption and a greater in-situ viscosity without any negative impact on injectivity. This plays a positive role in the oil displacement process. Moreover, the loss of viscosity in the effluent of polymer solutions was lower in low-salinity than in high-salinity brine, indicating that an increase in cations concentration (mainly driven by Ca2+ ions) has stronger effect on the viscosity of high-salinity polymer solution compared with low-salinity polymer.Keywords: polymer, heavy oil, low salinity, COBR interactions
Procedia PDF Downloads 931265 Assessment of Hypersaline Outfalls via Computational Fluid Dynamics Simulations: A Case Study of the Gold Coast Desalination Plant Offshore Multiport Brine Diffuser
Authors: Mitchell J. Baum, Badin Gibbes, Greg Collecutt
Abstract:
This study details a three-dimensional field-scale numerical investigation conducted for the Gold Coast Desalination Plant (GCDP) offshore multiport brine diffuser. Quantitative assessment of diffuser performance with regard to trajectory, dilution and mapping of seafloor concentration distributions was conducted for 100% plant operation. The quasi-steady Computational Fluid Dynamics (CFD) simulations were performed using the Reynolds averaged Navier-Stokes equations with a k-ω shear stress transport turbulence closure scheme. The study compliments a field investigation, which measured brine plume characteristics under similar conditions. CFD models used an iterative mesh in a domain with dimensions 400 m long, 200 m wide and an average depth of 24.2 m. Acoustic Doppler current profiler measurements conducted in the companion field study exhibited considerable variability over the water column. The effect of this vertical variability on simulated discharge outcomes was examined. Seafloor slope was also accommodated into the model. Ambient currents varied predominantly in the longshore direction – perpendicular to the diffuser structure. Under these conditions, the alternating port orientation of the GCDP diffuser resulted in simultaneous subjection to co-propagating and counter-propagating ambient regimes. Results from quiescent ambient simulations suggest broad agreement with empirical scaling arguments traditionally employed in design and regulatory assessments. Simulated dynamic ambient regimes showed the influence of ambient crossflow upon jet trajectory, dilution and seafloor concentration is significant. The effect of ambient flow structure and the subsequent influence on jet dynamics is discussed, along with the implications for using these different simulation approaches to inform regulatory decisions.Keywords: computational fluid dynamics, desalination, field-scale simulation, multiport brine diffuser, negatively buoyant jet
Procedia PDF Downloads 2141264 Performance Analysis of a Shell and Tube Heat Exchanger in the Organic Rankine Cycle Power Plant
Authors: Yogi Sirodz Gaos, Irvan Wiradinata
Abstract:
In the 500 kW Organic Rankine Cycle (ORC) power plant in Indonesia, an AFT (according to the Tubular Exchanger Manufacturers Association – TEMA) type shell and tube heat exchanger device is used as a pre-heating system for the ORC’s hot water circulation system. The pre-heating source is a waste heat recovery of the brine water, which is tapped from a geothermal power plant. The brine water itself has 5 MWₜₕ capacities, with average temperature of 170ᵒC, and 7 barg working pressure. The aim of this research is to examine the performance of the heat exchanger in the ORC system in a 500 kW ORC power plant. The data for this research were collected during the commissioning on the middle of December 2016. During the commissioning, the inlet temperature and working pressure of the brine water to the shell and tube type heat exchanger was 149ᵒC, and 4.4 barg respectively. Furthermore, the ΔT for the hot water circulation of the ORC system to the heat exchanger was 27ᵒC, with the inlet temperature of 140ᵒC. The pressure in the hot circulation system was dropped slightly from 7.4ᵒC to 7.1ᵒC. The flow rate of the hot water circulation was 80.5 m³/h. The presentation and discussion of a case study on the performance of the heat exchanger on the 500 kW ORC system is presented as follows: (1) the heat exchange duty is 2,572 kW; (2) log mean temperature of the heat exchanger is 13.2ᵒC; (3) the actual overall thermal conductivity is 1,020.6 W/m².K (4) the required overall thermal conductivity is 316.76 W/m².K; and (5) the over design for this heat exchange performance is 222.2%. An analysis of the heat exchanger detailed engineering design (DED) is briefly discussed. To sum up, this research concludes that the shell and tube heat exchangers technology demonstrated a good performance as pre-heating system for the ORC’s hot water circulation system. Further research need to be conducted to examine the performance of heat exchanger system on the ORC’s hot water circulation system.Keywords: shell and tube, heat exchanger, organic Rankine cycle, performance, commissioning
Procedia PDF Downloads 1431263 Synthesis, Crystal Structure Characterization, Hirshfeld Surface Analysis and Biological Activities of Two Schiff Base Polymorphs Derived From 2-Aminobenzonitrile
Authors: Nesrine Benarous, Hassiba Bougueria, Nabila Moussa Slimane, Aouatef Cherouana
Abstract:
Crystal polymorphism is important for the synthesis of more potent and bioactive pharmaceutical compounds, including their different properties, such as packing arrangement and conformation. In fact, polymorphism plays a vital role in drug development. Different parameters affect the crystallization and give their degree of freedom. Severalproperties affected polymorphism, like kinetics, thermodynamics, spectroscopy, and mechanical property. Various techniques are used for characterizing polymorphs, are crystallography, morphology, phase transitions, molecular motion, and chemical environment. In this work, crystal structures of two polymorphs (I and II) of the Schiff base (SB) title compound were prepared by condensation reaction. The crystal structures of both polymorphs were determined by single X-ray analysis. The two polymorphs crystallize in two different space groups: P21/c for I and Pbca for II. The dihedral angles between the two phenyl rings are 4.81º for I and 82.27º for II. Both crystal structures are built on the basis of moderate and weak hydrogen bonds, 𝜋-stacking, and halogen⋯halogeninteractions. On the other hand, Hirshfeld surface (HS) analysis indicates that the most important contributions to the crystal packing for the two polymorphs are from Cl⋯H/H⋯Cl, H⋯H, and N⋯H/H⋯N contacts. These are followed by C⋯H/H⋯C for compound I and C⋯C and by C⋯H/H⋯C contacts for compound II. Afterwards, the in vitro antibacterial activity revealed that the SB have been found effective against G- bacteria Klebsiella pneumonia andG+ bacteria Staphylococcus aureuswith MIC value of14.37μg/mL. Moreover, the SBexhibited moderate toxicity against Brine Shrimp with LC50 value of 44.19μg/mL.Keywords: polymorph, crystal structure, hirshfeld surface analysis, in vitro antibacterial activity, toxicity
Procedia PDF Downloads 1111262 Model Organic Ranikin Cycle Power Plant for Waste Heat Recovery in Olkaria-I Geothermal Power Plant
Authors: Haile Araya Nigusse, Hiram M. Ndiritu, Robert Kiplimo
Abstract:
Energy consumption is an indispensable component for the continued development of the human population. The global energy demand increases with development and population rise. The increase in energy demand, high cost of fossil fuels and the link between energy utilization and environmental impacts have resulted in the need for a sustainable approach to the utilization of the low grade energy resources. The Organic Rankine Cycle (ORC) power plant is an advantageous technology that can be applied in generation of power from low temperature brine of geothermal reservoirs. The power plant utilizes a low boiling organic working fluid such as a refrigerant or a hydrocarbon. Researches indicated that the performance of ORC power plant is highly dependent upon factors such as proper organic working fluid selection, types of heat exchangers (condenser and evaporator) and turbine used. Despite a high pressure drop, shell-tube heat exchangers have satisfactory performance for ORC power plants. This study involved the design, fabrication and performance assessment of the components of a model Organic Rankine Cycle power plant to utilize the low grade geothermal brine. Two shell and tube heat exchangers (evaporator and condenser) and a single stage impulse turbine have been designed, fabricated and the performance assessment of each component has been conducted. Pentane was used as a working fluid and hot water simulating the geothermal brine. The results of the experiment indicated that the increase in mass flow rate of hot water by 0.08 kg/s caused a rise in overall heat transfer coefficient of the evaporator by 17.33% and the heat transferred was increased by 6.74%. In the condenser, the increase of cooling water flow rate from 0.15 kg/s to 0.35 kg/s increased the overall heat transfer coefficient by 1.21% and heat transferred was increased by 4.26%. The shaft speed varied from 1585 to 4590 rpm as inlet pressure was varied from 0.5 to 5.0 bar and power generated was varying from 4.34 to 14.46W. The results of the experiments indicated that the performance of each component of the model Organic Rankine Cycle power plant operating at low temperature heat resources was satisfactory.Keywords: brine, heat exchanger, ORC, turbine
Procedia PDF Downloads 6491261 In Vitro Study of Antioxidant Capacity of Chrysanthemum Indicum Extract
Authors: Puchita Chokcharoenying
Abstract:
Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Antioxidants are substances found in medicinal plants to help prevent heart disease, stroke, and some cancers. This study focused on evaluating the flavonoids content of Chrysanthemum Indicum and determine their antioxidant capacity by using DPPH and ABTS radical scavenging capacity assay. The total flavonoid content of C. indicumextract was determined and expressed as quercetin equivalents (QE)/g measured by an aluminiumchloride colorimetric method. The results showed that the IC50 of C. indicum extract were 83.57μg/mL ± 0.875 and52.57μg/mL ± 0.632for DPPH and ABTS, respectively. C. indicumextract exhibited antioxidant activities as a concentration dependent manner. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In summary, C. indicum extract is rich in flavonoids, which have potent antioxidant properties. Thus, C. indicum extract is a good source of antioxidants and can be developed for medicinal purposes. Nevertheless, more research on the antioxidant activity of C. indicum extract and in vivo antioxidant studies are still needed.Keywords: ABTS assay, antioxidant, chrysanthemum indicum, DPPH assay, total flavonoid content
Procedia PDF Downloads 2581260 Performance of Non-toxic, Corrosion Resistant, and Lubricious Metalworking Fluids under Machining
Authors: Ajay Pratap Singh Lodhi, Deepak Kumar
Abstract:
Vegetable oil-based environmentally friendly metalworking fluids (MWFs) are formulated. The tribological performance, cytotoxicity, and corrosion resistance of the formulated fluids (FFs) are evaluated and benchmarked with commercial mineral oil-based MWFs (CF). Results show that FFs exhibited better machining characteristics (roughness, cutting forces, and surface morphology) during machining than CF. MTT assay and Live dead cell assay confirm the cytocompatibility nature of the FFs relative to the toxic CF. Electrochemical analysis shows that FFs and CF exhibited comparable corrosion current density.Keywords: corrosion inhibitors, cytotoxicity, machining, MTT assay, Taguchi method, vegetable oil
Procedia PDF Downloads 1881259 Utility of the Loop-Mediated Isothermal Amplification Assay for the Diagnosis of Visceral Leishmaniasis from Blood Samples in Ethiopia
Authors: Dawit Gebreegzabher Hagos, Yazezew Kebede Kiro, Mahmud Abdulkader, Henk H. D. F. Schallig, Dawit Wolday
Abstract:
Rapid and accurate visceral leishmaniasis (VL) diagnosis is needed to initiate prompt treatment to reduce morbidity and mortality. Here, we evaluated the performance of loop-mediated isothermal amplification (LAMP) assay for the diagnosis of VL from blood in an endemic area in Ethiopia. LAMP was positive in 117/122 confirmed VL cases and negative in 149/152 controls, resulting in a sensitivity of 95.9% (95% CI: 90.69–98.66) and a specificity of 98.0% (95% CI: 94.34–99.59), respectively. The sensitivity of the LAMP assay was 95.0% (95% CI: 88.61–98.34) in HIV-negatives and 100% (95% CI: 85.18–100.0) in HIV-positives. Compared with microscopy, LAMP detected 82/87 (94.3%, 95% CI: 87.10–98.11) of the microscopy1 cases and was negative in 11/27 (40.7%, 95% CI: 22.39–61.20) of the microscopy2 cases. Compared with the rK39 serology, LAMP detected 113/120 (94.2%, 95% CI: 88.35–97.62) of the rK391 cases and was negative in 149/154 (96.8%, 95% CI: 92.59–98.94) of the rK392 cases. However, when compared with microscopy only, rK39 detected 83/87 (95.4%, 95% CI: 88.64–98.73) of the microscopy1 cases and negative in only 12/27 (44.4%, 95% CI: 25.48–64.67) of the microscopy– cases. There was an excellent agreement between rK39 and LAMP (Kappa 5 0.91, 95% CI: 0.86–0.96). Furthermore, an algorithm using rK39 followed by LAMP would yield a sensitivity of 99.2% (95%CI: 95.52–99.89) and a specificity of 98.0% (95% CI: 94.34–99.59). The findings demonstrate that the LAMP assay is an accurate and rapid molecular assay for VL diagnosis, including in HIV-1 co-infected patients, in an endemic setting.Keywords: visceral leishmaniasis, HIV, diagnosis, LAMP, Ethiopia
Procedia PDF Downloads 981258 Advanced Lithium Recovery from Brine: 2D-Based Ion Selectivity Membranes
Authors: Nour S. Abdelrahman, Seunghyun Hong, Hassan A. Arafat, Daniel Choi, Faisal Al Marzooqi
Abstract:
Abstract—The advancement of lithium extraction methods from water sources, particularly saltwater brine, is gaining prominence in the lithium recovery industry due to its cost-effectiveness. Traditional techniques like recrystallization, chemical precipitation, and solvent extraction for metal recovery from seawater or brine are energy-intensive and exhibit low efficiency. Moreover, the extensive use of organic solvents poses environmental concerns. As a result, there's a growing demand for environmentally friendly lithium recovery methods. Membrane-based separation technology has emerged as a promising alternative, offering high energy efficiency and ease of continuous operation. In our study, we explored the potential of lithium-selective sieve channels constructed from layers of 2D graphene oxide and MXene (transition metal carbides and nitrides), integrated with surface – SO₃₋ groups. The arrangement of these 2D sheets creates interplanar spacing ranging from 0.3 to 0.8 nm, which forms a barrier against multivalent ions while facilitating lithium-ion movement through nano capillaries. The introduction of the sulfonate group provides an effective pathway for Li⁺ ions, with a calculated binding energy of Li⁺ – SO³⁻ at – 0.77 eV, the lowest among monovalent species. These modified membranes demonstrated remarkably rapid transport of Li⁺ ions, efficiently distinguishing them from other monovalent and divalent species. This selectivity is achieved through a combination of size exclusion and varying binding affinities. The graphene oxide channels in these membranes showed exceptional inter-cation selectivity, with a Li⁺/Mg²⁺ selectivity ratio exceeding 104, surpassing commercial membranes. Additionally, these membranes achieved over 94% rejection of MgCl₂.Keywords: ion permeation, lithium extraction, membrane-based separation, nanotechnology
Procedia PDF Downloads 711257 Deproteination and Demineralization of Shrimp Waste Using Lactic Acid Bacteria for the Production of Crude Chitin and Chitosan
Authors: Farramae Francisco, Rhoda Mae Simora, Sharon Nunal
Abstract:
Deproteination and demineralization efficiencies of shrimp waste using two Lactobacillus species treated with different carbohydrate sources for chitin production, its chemical conversion to chitosan and the quality of chitin and chitosan produced were determined. Using 5% glucose and 5% cassava starch as carbohydrate sources, pH slightly increased from the initial pH of 6.0 to 6.8 and 7.2, respectively after 24 h and maintained their pH at 6.7 to 7.3 throughout the treatment period. Demineralization (%) in 5 % glucose and 5 % cassava was highest during the first day of treatment which was 82% and 83%, respectively. Deproteination (%) was highest in 5% cassava starch on the 3rd day of treatment at 84.4%. The obtained chitin from 5% cassava and 5% glucose had a residual ash and protein below 1% and solubility of 59% and 44.3%, respectively. Chitosan produced from 5% cassava and 5% glucose had protein content below 0.05%; residual ash was 1.1% and 0.8%, respectively. Chitosan solubility and degree of deacetylation were 56% and 33% in 5% glucose and 48% and 29% in 5% cassava, respectively. The advantage this alternative technology offers over that of chemical extraction is large reduction in chemicals needed thus less effluent production and generation of a protein-rich liquor, although the demineralization process should be improved to achieve greater degree of deacetylation.Keywords: alternative carbon source, bioprocessing, lactic acid bacteria, waste utilization
Procedia PDF Downloads 4851256 Evaluation Of In Vitro Antioxidant Potential of Camellia Sinensis Leaves Extract
Authors: Jirathan Pongchababnapa
Abstract:
Polyphenols are the most common antioxidant found in plants and are efficient in capturing oxidative free radicals. Antioxidants are substances found in medicinal plants which may have a protective role to play in certain conditions such as heart disease, stroke and some cancers. By relying on these benefits, we have traced out the presence of antioxidant in Camellia sinensis leaves extract. This study aims to evaluate flavonoids content in C. sinensisextract and investigate antioxidant activities by using DPPH and ABTS radical scavenging capacity assay. The total flavonoid content of C. Sinensis extract was determined and expressed as quercetin equivalents (QE)/g measured by the aluminum chloride colorimetric method. The results showed that the IC₅₀ of C. Sinensis leaves extract were 40.90 μg/mL ± 0.755 and32.96 μg/mL ± 0.679 for DPPH and ABTS, respectively. C. Sinensis extract at increasing concentration showed antioxidant activities as a concentration dependent manner. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. Sinensis extract consisted of a high amount of flavonoids content which possesses potent antioxidant activity. However, further investigation on the identification of pure compound of this plant and molecular antioxidant assays are still required.Keywords: ABTS assay, antioxidant, camellia sinensis, DPPH assay, total flavonoid content
Procedia PDF Downloads 2101255 Studies on the Recovery of Calcium and Magnesium from Red Seawater by Nanofiltration Membrane
Authors: Mohamed H. Sorour, Hayam F. Shaalan, Heba A. Hani, Mahmoud A. El-Toukhy
Abstract:
This paper reports the results of nanofiltration (NF) polymeric membrane for the recovery of divalent ions (calcium and magnesium) from Red Seawater. Pilot plant experiments have been carried out using Alfa-Laval (NF 2517/48) membrane module. System was operated in both total recirculation mode (permeate and brine) and brine recirculation mode under hydraulic pressure of 15 bar. Impacts of some chelating agents on both flux and rejection have been also investigated. Results indicated that pure water permeability ranges from 17 to 85.5 L/m²h at 2-15 bar. Comparison with seawater permeability under the same operating pressure values reveals lower values of 8.9-31 L/m²h manifesting the effect of the osmotic pressure of seawater. Overall total dissolved solids (TDS) reduction was almost constant without incorporation of chelating agents. On the contrary of expectations, the use of chelating agents N-(2-hydroxyethyl) ethylene diamine-N,N´,N´-triacetic acid (HEDTA) and ethylene glycol bis (2-aminoethyl ether)-N,N,N´,N´-tetraacetic acid (EGTA) showed flux decline of about 3-15%. Analysis of rejection data of total recirculation mode showed reasonable rejection values of 35%, 59% and 90% for Ca, Mg and SO₄, respectively. Operating under brine recirculation mode only showed a decrease of rejection to 33%, 56% and 86% for Ca, Mg and SO₄, respectively. The use of chelating agents has no substantial effect on NF membrane performance except for increasing the total Ca rejection to 48 and 65% for EGTA and HEDTA, respectively. Results, in general, confirmed the powerful separation of NF technology for softening and recovery of divalent ions from seawater. It is anticipated that increasing operating pressure beyond the limits of our investigations would improve the rejection and flux values. A trade-off should be considered between operating cost (due to higher pressure and marginal benefits as manifested by expected improved performance). The experimental results fit well with the formulated rejection empirical correlations and the published ones.Keywords: nanofiltration, seawater, recovery, calcium, magnesium
Procedia PDF Downloads 1651254 Invitro Study of Anti-Leishmanial Property of Nigella Sativa Methanalic Black Seed Extract
Authors: Tawqeer Ali Syed, Prakash Chandra
Abstract:
This study aims to evaluate the antileishmanial activity of Nigella sativa black seed extract. This well-known plant extract was taken from the botanical garden of Kashmir. Materials and Methods: The methanolic extracts of these plants were screened for their antileishmanial activity against Leishmania major using 3‑(4.5‑dimethylthiazol‑2yl)‑2.5‑diphenyltetrazolium bromide assay or MTT assay. Results: The methanolic extract of Nigella sativa showed potential antileishmanial activity at an inhibition% value of 80.29% ± 0.65%. IC 50 was calculated after 48 hours to be 964.3 µg/ml. Conclusion: Considering these results, these medicinal plants from Kashmir could serve as potential drug sources for antileishmanial compounds.Keywords: MTT assay, antileishmanial, cell viability, Nigella sativa
Procedia PDF Downloads 2121253 Silver Nanoparticles Synthesized in Plant Extract Against Acute Hepatopancreatic Necrosis of Shrimp: Estimated By Multiple Models
Authors: Luz del Carmen Rubí Félix Peña, Jose Adan Felix-Ortiz, Ely Sara Lopez-Alvarez, Wenceslao Valenzuela-Quiñonez
Abstract:
On a global scale, Mexico is the sixth largest producer of farmed white shrimp (Penaeus vannamei). The activity suffered significant economic losses due to acute hepatopancreatic necrosis (AHPND) caused by a strain of Vibrio parahaemolyticus. For control, the first option is the application of antibiotics in food, causing changes in the environment and bacterial communities, which has produced greater virulence and resistance of pathogenic bacteria. An alternative treatment is silver nanoparticles (AgNPs) generated by green synthesis, which have shown an antibacterial capacity by destroying the cell membrane or denaturing the cell. However, the doses at which these are effective are still unknown. The aim is to calculate the minimum inhibitory concentration (MIC) using the Gompertz, Richard, and Logistic model of biosynthesized AgNPs against a strain of V. parahaemolyticus. Through the testing of different formulations of AgNPs synthesized from Euphorbia prostrate (Ep) extracts against V. parahaemolyticus causing AHPND in white shrimp. Aqueous and ethanol extracts were obtained, and the concentration of phenols and flavonoids was quantified. In the antibiograms, AgNPs were formulated in ethanol extracts of Ep (20 and 30%). The inhibition halo at well dilution test were 18±1.7 and 17.67±2.1 mm against V. parahaemolyticus. A broth microdilution was performed with the inhibitory agents (aqueous and ethanolic extracts and AgNPs) and 20 μL of the inoculum of V. parahaemolyticus. The MIC for AgNPs was 6.2-9.3 μg/mL and for ethanol extract of 49-73 mg/mL. The Akaike index (AIC) was used to choose the Gompertz model for ethanol extracts of Ep as the best data descriptor (AIC=204.8, 10%; 45.5, 20%, and 204.8, 30%). The Richards model was at AgNPs ethanol extract with AIC=-9.3 (10%), -17.5 (20 and 30%). The MIC calculated for EP extracts with the modified Gompertz model were 20 mg/mL (10% and 20% extract) and 40 mg/mL at 30%, while Richard was winner for AgNPs-synthesized it was 5 μg/mL (10% and 20%) and 8 μg/mL (30%). The solver tool Excel was used for the calculations of the models and inhibition curves against V.parahaemolyticus.Keywords: green synthesis, euphorbia prostata, phenols, flavonoids, bactericide
Procedia PDF Downloads 1061252 Optimizing Fermented Paper Production Using Spyrogira sp. Interpolating with Banana Pulp
Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn
Abstract:
Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.Keywords: cellulose, fermentation, grammage, paper, Spirogyra sp.
Procedia PDF Downloads 3331251 In vitro Antioxidant Activity of Caesalpinia sappan Extract
Authors: Monthon Tangjitmungman
Abstract:
Numerous diseases have been linked to oxidative stress, in which a disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Due to the presence of phenolic compounds in Caesalpinia sappan has been discovered to have antioxidant activity. It has several health benefits, the most important of which is preventing cardiovascular and cancer diseases. This study aimed to determine the phenolic content and antioxidant activity of C. sappan extract using a variety of antioxidant assays. The extract of C. sappan was made using a mixture of solvents (ethyl alcohol: water in ratio 8:2). The total phenolic content of C. sappan extract was determined and expressed as gallic acid equivalents using the Folin-Cioucalteu method (GAE). The antioxidant activity of C. sappan extract was assessed using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and the ABTS radical scavenging capacity assay. An association was found between antioxidant activity and total phenol content. The antioxidant activity of C. sappan extract was also determined by DPPH and ABTS assays. The IC50 values for C. sappan extract from DPPH and ABTS assays were 54.48 μg/mL ± 0.545 and 25.46 μg/mL ± 0.790, respectively, in the DPPH assay. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. sappan extract contains a high level of total phenolics and exhibits significant antioxidant activity. Nevertheless, more research should be done on the antioxidant activity, such as SOD and ROS scavenging assays and in vivo experiments, to determine whether the compound has antioxidant activity.Keywords: ABTS assay, antioxidant activity, Caesalpinia sappan, DPPH assays, total phenolic content
Procedia PDF Downloads 3841250 Investigation the Effect of Quenching Media on Abrasive Wear in Grade Medium Carbon Steel
Authors: Abbas S. Alwan, Waleed K. Hussan
Abstract:
In this paper, a general verification of possible heat treatment of steel has been done with the view of conditions of real abrasive wear of rotivater with soil texture. This technique is found promising to improve the quality of agriculture components working with the soil in dry condition. Abrasive wear resistance is very important in many applications and in most cases it is directly correlated with the hardness of materials surface. Responded of heat treatments were carried out in various media (Still air, Cottonseed oil, and Brine water 10 %) and follow by low-temperature tempering (250°C) was applied on steel type (AISI 1030). After heat treatment was applied wear with soil texture by using tillage process to determine the (actual wear rate) of the specimens depending on weight loss method. It was found; the wear resistance Increases with increase hardness with varying quenching media as follows; 30 HRC, 45 HRC, 52 HRC, and 60 HRC for nontreated (as received) cooling media as still air, cottonseed oil, and Brine water 10 %, respectively. Martensitic structure with retained austenite can be obtained depending on the quenching medium. Wear was presented on the worn surfaces of the steels which were used in this work.Keywords: microstructures, hardness, abrasive wear, heat treatment, soil texture
Procedia PDF Downloads 3881249 Defense Priming from Egg to Larvae in Litopenaeus vannamei with Non-Pathogenic and Pathogenic Bacteria Strains
Authors: Angelica Alvarez-Lee, Sergio Martinez-Diaz, Jose Luis Garcia-Corona, Humberto Lanz-Mendoza
Abstract:
World aquaculture is always looking for improvements to achieve productions with high yields avoiding the infection by pathogenic agents. The best way to achieve this is to know the biological model to create alternative treatments that could be applied in the hatcheries, which results in greater economic gains and improvements in human public health. In the last decade, immunomodulation in shrimp culture with probiotics, organic acids and different carbon sources has gained great interest, mainly in larval and juvenile stages. Immune priming is associated with a strong protective effect against a later pathogen challenge. This work provides another perspective about immunostimulation from spawning until hatching. The stimulation happens during development embryos and generates resistance to infection by pathogenic bacteria. Massive spawnings of white shrimp L. vannamei were obtained and placed in experimental units with 700 mL of sterile seawater at 30 °C, salinity of 28 ppm and continuous aeration at a density of 8 embryos.mL⁻¹. The immunostimulating effect of three death strains of non-pathogenic bacterial (Escherichia coli, Staphylococcus aureus and Bacillus subtilis) and a pathogenic strain for white shrimp (Vibrio parahaemolyticus) was evaluated. The strains killed by heat were adjusted to O.D. 0.5, at A 600 nm, and directly added to the seawater of each unit at a ratio of 1/100 (v/v). A control group of embryos without inoculum of dead bacteria was kept under the same physicochemical conditions as the rest of the treatments throughout the experiment and used as reference. The duration of the stimulus was 12 hours, then, the larvae that hatched were collected, counted and transferred to a new experimental unit (same physicochemical conditions but at a salinity of 28 ppm) to carry out a challenge of infection against the pathogen V. parahaemolyticus, adding directly to seawater an amount 1/100 (v/v) of the live strain adjusted to an OD 0.5; at A 600 nm. Subsequently, 24 hrs after infection, nauplii survival was evaluated. The results of this work shows that, after 24 hrs, the hatching rates of immunostimulated shrimp embryos with the dead strains of B. subtillis and V. parahaemolyticus are significantly higher compared to the rest of the treatments and the control. Furthermore, survival of L. vanammei after a challenge of infection of 24 hrs against the live strain of V. parahaemolyticus is greater (P < 0.05) in the larvae immunostimulated during the embryonic development with the dead strains B. subtillis and V. parahaemolyticus, followed by those that were treated with E. coli. In summary superficial antigens can stimulate the development cells to promote hatching and can have normal development in agreeing with the optical observations, plus exist a differential response effect between each treatment post-infection. This research provides evidence of the immunostimulant effect of death pathogenic and non-pathogenic bacterial strains in the rate of hatching and oversight of shrimp L. vannamei during embryonic and larval development. This research continues evaluating the effect of these death strains on the expression of genes related to the defense priming in larvae of L. vannamei that come from massive spawning in hatcheries before and after the infection challenge against V. parahaemolyticus.Keywords: immunostimulation, L. vannamei, hatching, survival
Procedia PDF Downloads 1421248 Novel Nanomagnetic Beads Based- Latex Agglutination Assay for Rapid Diagnosis of Human Schistosomiasis Haematobium
Authors: Ibrahim Aly, Rabab Zalat, Bahaa EL Deen W. El Aswad, Ismail M. Moharm, Basam M. Masoud, Tarek Diab
Abstract:
The objective of the present study was to evaluate the novel nanomagnetic beads based–latex agglutination assay (NMB-LAT) as a simple test for diagnosis of S. haematobium as well as standardize the novel nanomagnetic beads based –ELISA (NMB-ELISA). According to urine examination this study included 85 S. haematobium infected patients, 30 other parasites infected patients and 25 negative control samples. The sensitivity of novel NMB-LAT was 82.4% versus 96.5% and 88.2% for NMB-ELISA and currently used sandwich ELISA respectively. The specificity of NMB-LAT was 83.6% versus 96.3% and 87.3% for NMB-ELISA and currently used sandwich ELISA respectively. In conclusion, the novel NMB-ELISA is a valuable applicable diagnostic technique for diagnosis of human schistosomiasis haematobium. The novel NMB-ELISA assay is a suitable applicable diagnostic method in field survey especially when followed by ELISA as a confirmatory test in query false negative results. Trials are required to increase the sensitivity and specificity of NMB-ELISA assay.Keywords: diagnosis, iatex agglutination, nanomagnetic beads, sandwich ELISA
Procedia PDF Downloads 3821247 Prevalence and Evaluation of Antimicrobial Activity of Dodonaea viscosa Extract and Antibacterial Agents against Salmonella spp. Isolated from Poultry
Authors: Shayma Munqith Al-Baker, Fadhl Ahmed Saeed Al-Gasha’a, Samira Hamid Hanash, Ahmed Ali Al-Hazmi
Abstract:
A total of 200 samples (180 fecal materials and 20 organ samples) were collected from (5 different poultry farms, 10 local poultry shops, 5 houses poultry, 5 Eggs stores shops and 5 hand slaughters centers) in Ibb city, Yemen, 2014. According to morphological, cultural, as well as biochemical characterization and serological tests, 59 29.5% isolates were identified as Salmonella spp. and all Salmonella isolates were categorized by serotype, which comprised of, 37 62.71% Salmonella Typhimurium serovar, 21 35.59%. Salmonella Enteritidis serovar and 11.69% Salmonella Heidelberg serovar. Antibiotic sensitivity test was done for bacterial isolates and the results showed there were clear differences in antibiotic resistant. Antimicrobial susceptibility of the isolates varies as follows: Ofloxacin 79.66%, Ciprofloxacin 67.80%, Colistin 59.32% and Gentamycin 52.54%. All of isolates were resistant to Erythromycin, Penicillin and Lincomycin. Antibacterial activity was done for both aqueous and ethanol extracts of Dodonaea viscosa plant by using well and disc diffusion assay. The results indicated that well diffusion assay had best results than disc diffusion assay, the highest inhibition zone was 22 mm for well diffusion and 15 mm for disc diffusion assay, the results observed that ethanol extract had best antibacterial effect than aqueous extract which the percentage of bacterial isolates affected with ethanol extract was 71.19% comparing with aqueous extract 28.81% by using disc diffusion assay, while the percentage of bacterial isolates affected with ethanol extract was 88.13% comparing with aqueous extract 52.54% by using will diffusion assay.Keywords: Salmonella spp, Dodonaea viscosa, antimicrobial and salmonellosis
Procedia PDF Downloads 4731246 Mutations in rpoB, katG and inhA Genes: The Association with Resistance to Rifampicin and Isoniazid in Egyptian Mycobacterium tuberculosis Clinical Isolates
Authors: Ayman K. El Essawy, Amal M. Hosny, Hala M. Abu Shady
Abstract:
The rapid detection of TB and drug resistance, both optimizes treatment and improves outcomes. In the current study, respiratory specimens were collected from 155 patients. Conventional susceptibility testing and MIC determination were performed for rifampicin (RIF) and isoniazid (INH). Genotype MTBDRplus assay, which is a molecular genetic assay based on the DNA-STRIP technology and specific gene sequencing with primers for rpoB, KatG, and mab-inhA genes were used to detect mutations associated with resistance to rifampicin and isoniazid. In comparison to other categories, most of rifampicin resistant (61.5%) and isoniazid resistant isolates (47.1%) were from patients relapsed in treatment. The genotypic profile (using Genotype MTBDRplus assay) of multi-drug resistant (MDR) isolates showed missing of katG wild type 1 (WT1) band and appearance of mutation band katG MUT2. For isoniazid mono-resistant isolates, 80% showed katG MUT1, 20% showed katG MUT1, and inhA MUT1, 20% showed only inhA MUT1. Accordingly, 100% of isoniazid resistant strains were detected by this assay. Out of 17 resistant strains, 16 had mutation bands for katG distinguished high resistance to isoniazid. The assay could clearly detect rifampicin resistance among 66.7% of MDR isolates that showed mutation band rpoB MUT3 while 33.3% of them were considered as unknown. One mono-resistant rifampicin isolate did not show rifampicin mutation bands by Genotype MTBDRplus assay, but it showed an unexpected mutation in Codon 531 of rpoB by DNA sequence analysis. Rifampicin resistance in this strain could be associated with a mutation in codon 531 of rpoB (based on molecular sequencing), and Genotype MTBDRplus assay could not detect the associated mutation. If the results of Genotype MTBDRplus assay and sequencing were combined, this strain shows hetero-resistance pattern. Gene sequencing of eight selected isolates, previously tested by Genotype MTBDRplus assay, could detect resistance mutations mainly in codon 315 (katG gene), position -15 in inhA promotes gene for isoniazid resistance and codon 531 (rpoB gene) for rifampicin resistance. Genotyping techniques allow distinguishing between recurrent cases of reinfection or reactivation and supports epidemiological studies.Keywords: M. tuberculosis, rpoB, KatG, inhA, genotype MTBDRplus
Procedia PDF Downloads 1661245 Bacterial Profiling and Development of Molecular Diagnostic Assays for Detection of Bacterial Pathogens Associated with Bovine mastitis
Authors: Aqeela Ashraf, Muhammad Imran, Tahir Yaqub, Muhammad Tayyab, Yung Fu Chang
Abstract:
For the identification of bovine mastitic pathogen, an economical, rapid and sensitive molecular diagnostic assay is developed by PCR multiplexing of gene and pathogenic species specific DNA sequences. The multiplex PCR assay is developed for detecting nine important bacterial pathogens causing mastitis Worldwide. The bacterial species selected for this study are Streptococcus agalactiae, Streptococcus dysagalactiae, Streptococcus uberis, Staphylococcus aureus, Escherichia coli, Staphylococcus haemolyticus, Staphylococcus chromogenes Mycoplasma bovis and Staphylococcus epidermidis. A single reaction assay was developed and validated by 27 reference strains and further tested on 276 bacterial strains obtained from culturing mastitic milk. The multiplex PCR assay developed here is further evaluated by applying directly on genomic DNA isolated from 200 mastitic milk samples. It is compared with bacterial culturing method and proved to be more sensitive, rapid, economical and can specifically identify 9 bacterial pathogens in a single reaction. It has detected the pathogens in few culture negative mastitic samples. Recognition of disease is the foundation of disease control and prevention. This assay can be very helpful for maintaining the udder health and milk monitoring.Keywords: multiplex PCR, bacteria, mastitis, milk
Procedia PDF Downloads 330