Haitian Variant Vibrio cholerae O1 Strains Manifest Higher Virulence in Animal Models

Vibrio cholerae causes fatal diarrheal disease cholera in humans due to consumption of contaminated water and food. To instigate the disease, the bacterium must evade the host intestinal innate immune system; penetrate the mucus layer of the small intestine, adhere and multiply on the surface of microvilli and produce toxin(s) through the action of virulence associated genes. V. cholerae O1 that has caused a major cholera outbreak in Haiti contained several unique genetic signatures. These novel traits are used to differentiate them from the canonical El Tor strains. Several studies reported the spread of these Haitian variant strains in different parts of the world including Asia and Africa, but there is a paucity of information on the clinical consequence of these genetic changes. To understand the impact of these changes, we undertook a study involving mice and rabbit models to evaluate the pathogenesis. The colonization ability of Haitian variant strain in comparison to canonical El Tor strain was found to be significantly more in both suckling mice and rabbit model. Adult mice also displayed the same results. Besides that, infection patterns of Haitian variant strains showed a completely different picture. Increased mucosal damaging, colonization, and inflammatory changes were observed through hematoxylin-eosin staining and transmission electron microscopy. Fluid accumulation ability was also significantly higher in rabbit model. Our study indicated that these virulence features of the Haitian variant strain may have some association with the severe clinical outcome of the cholera patients in different parts of the world

Characterization of Vibrio cholerae O1 strains that trace the origin of Haitian-like genetic traits

Vibrio cholerae O1 is the etiological agent of the severe diarrheal disease cholera. The bacterium has recently been causing outbreaks in Haiti with catastrophic effects. Numerous mutations have been reported in V. cholerae O1 strains associated with the Haitian outbreak. These mutations encompass among other the genes encoding virulence factors such as the pilin subunit of the toxin-co-regulated pilus (tcpA), cholera toxin B subunit (ctxB), repeat in toxins (rtxA), and other genes such as the quinolone resistance-determining region (QRDR) of gyrase A (gyrA), rstB of RS element along with the alteration in the number of repeat sequences at the promoter region of ctxAB. Given the numerous genetic changes in those Haitian isolates, we decided to investigate the possible origins of those variations in the Indian subcontinent. Thus, we determined the genetic traits among V. cholerae O1 strains in Delhi, India. A total of 175 strains isolated from cholera patients during 2004 to 2012 were analysed in the present study. Our results showed that all the tested strains carried Haitian type tcpA (tcpACIRS) and variant gyrA indicating their first appearance before 2004 in Delhi. The Haitian variant rtxA and ctxB7 were first detected in Delhi during 2004 and 2006, respectively. Interestingly, not a single strain with the combination of El Tor rtxA and ctxB7 was detected in this study. The Delhi strains carried four heptad repeats (TTTTGAT) in the CT promoter region whereas Haitian strains carried 5 such repeats. Delhi strains did not have any deletion mutations in the rstB like Haitian strains. Overall, our study demonstrates the sequential accumulation of Haitian-like genetic traits among V. cholerae O1 strains in Delhi at different time points prior to the Haitian cholera outbreak

Evaluation of primary phase morphology of cooling slope cast Al-Si-Mg alloy samples using image texture analysis

Rheopressure die casting is one of the newest casting processes of present era for manufacturing of near-net shaped cast components with improved mechanical properties and high dimensional accuracy. Rheopressure die casting demands especially prepared semi-solid alloy slurry having nearly globular primary Al phase. In this study, a cooling slope has been employed to produce semi-solid slurry of Al-Si-Mg (A356) alloy and successively cast in a metallic mould. Image texture analysis techniques have been implemented for accurate evaluation of the primary phase morphology of cast samples. In this research, efforts have been made to apply fractal analysis and run-length statistical analysis techniques for automatic characterization of optical micrographs of cast samples produced at different processing conditions

Studies on die filling of A356 Al alloy and development of a steering knuckle component using rheo pressure die casting system

In this study, a computational fluid dynamics (CFD) model is developed to investigate die filling of semi solid slurry as part of rheo pressure die casting (RPDC) system. The die filling cavity corresponds to that of an automobile steering knuckle, and the slurry is made of A356 aluminium alloy. The rheological model used in the CFD simulation is determined experimentally. The results obtained from present numerical model includes flow field of the slurry within the die cavity, viscosity evolution, solid fraction distribution, temperature and pressure distribution during solidification within cavity during die filling stage. The main objective of the study is to determine the gating arrangement, pouring temperature, and injection conditions for desirable microstructure and mechanical properties of the developed component. To study the effect of injection conditions on die filling capability of the said alloy slurry, five injection profiles are studied, with a variation in final injection velocity between 2–3.2 m/s. In order to corroborate the findings of the present study, microstructural morphology and structure-property correlation have been studied, primarily in the form of optical microscopy and macro hardness measurements, by obtaining samples from different locations of the solidified component

Effect of pouring temperature on cooling slope casting of semi-solid Al-Si-Mg alloy

Present trend of semi-solid processing is directed towards rheocasting route which allows manufacturing of near-net-shape cast components directly from the prepared semi-solid slurry. Generation of globular equi-axed grains during solidification of rheocast components, compared to the columnar dendritic structure of conventional casting routes, facilitates the manufacturing of components with improved mechanical properties and structural integrity. In the present investigation, a cooling slope has been designed and indigenously fabricated to produce semi solid slurry of Al-Si-Mg (A356) alloy and successively cast in a metallic mould. The scope of the present work discusses about development of a numerical model to simulate the liquid metal flow through cooling slope using Eulerian two-phase flow approach and to investigate the effect of pouring temperature on cooling slope semi-solid slurry generation process. The two phases considered in the present model are liquid metal and air. Solid fraction evolution of the solidifying melt is tracked at different locations of the cooling slope, following Schiel's equation. The continuity equation, momentum equation and energy equation are solved considering thin wall boundary condition approach. During solidification of the liquid metal, a modified temperature recovery scheme has been employed taking care of the latent heat release and change of fraction of liquid. The results obtained from simulations are compared with experimental findings and good agreement has been found

Phase Field Simulation of Equiaxed Microstructure Formation during Semi-solid Processing of A380 Al Alloy

A phase field modelling approach is implemented in the present study towards simulation of microstructure evolution during cooling slope semi solid slurry generation process of A380 Aluminium alloy. First, experiments are performed to evaluate the number of seeds required within the simulation domain to simulate near spherical microstructure formation, occurs during cooling slope processing of the melt. Subsequently, microstructure evolution is studied employing a phase field method. Simulations are performed to understand the effect of cooling rate on the slurry microstructure. Encouraging results are obtained from the simulation studies which are validated by experimental observations. The results obtained from mesoscopic phase field simulations are grain size, grain density, degree of sphericity of the evolving primary Al phase and the amount of solid fraction present within the slurry at different time frames. Effect of grain refinement also has been studied with an aim of improving the slurry microstructure further. Insight into the process has been obtained from the numerical findings, which are found to be useful for process control

Optimization of degree of sphericity of primary phase during cooling slope casting of A356 Al alloy: Taguchi method and regression analysis

The present work presents the results of experimental investigation of semi-solid rheocasting of A356 Al alloy using a cooling slope. The experiments have been carried out following Taguchi method of parameter design (orthogonal array of L9 experiments). Four key process variables (slope angle, pouring temperature, wall temperature, and length of travel of the melt) at three different levels have been considered for the present experimentation. Regression analysis and analysis of variance (ANOVA) has also been performed to develop a mathematical model for degree of sphericity evolution of primary a-Al phase and to find the significance and percentage contribution of each process variable towards the final outcome of degree of sphericity, respectively. The best processing condition has been identified for optimum degree of sphericity (0.83) as A3, B3, C2, D1 i.e., slope angle of 60 �, pouring temperature of 650 �C, wall temperature 60 �C, and 500 mm length of travel of the melt, based on mean response and signal to noise ratio (SNR). ANOVA results shows that the length of travel has maximum impact on degree of sphericity evolution. The predicted sphericity obtained from the developed regression model and the values obtained experimentally are found to be in good agreement with each other. The sphericity values obtained from confirmation experiment, performed at 95% confidence level, ensures that the optimum result is correct and also the confirmation experiment values are within permissible limits

Microstructure Evolution and Rheological Behavior of Cooling Slope Processed Al-Si-Cu-Fe Alloy Slurry

In the present work, microstructure evolution during semi-solid slurry generation of Al-Si-Cu-Fe alloy, using a cooling slope, was studied and the effect of microstructural morphology of the slurry on its rheological behavior was investigated. Microstructure evolution during melt flow along the slope was studied by extracting samples from various locations of the slope and performing rapid oil quenching experiments. Quantitative investigation was performed to evaluate primary phase shape and size for different process conditions of the semi-solid slurry, and subsequently rheological investigations were performed to correlate slurry morphology with its flow behavior. Three different types of rheological experiments were performed: isothermal test, shear jump test, and shear time test, in order to investigate rheological behavior of the semi-solid slurry. In addition, effect of melt treatment, by adding modifier (0.1 wt pct of Al-10Sr) and grain refiner (0.15 wt pct of Al-5Ti-1B), on the microstructure evolution during slurry generation, flow behavior of the slurry, and intermetallics formation was studied

Die Filling Behaviour of Semi Solid A356 Al Alloy Slurry During Rheo Pressure Die Casting

The present work discusses the findings obtained from simulations of semi solid die filling of a steering knuckle, prior to actual component development using in-house developed rheo pressure die casting system. Die filling capability of A356 Al alloy at semi-solid state has been investigated using commercial software Flow-3Dcast to optimise the pouring temperature of semi-solid slurry into the die cavity, while all other variables such as gating design, die preheat temperature and injection velocity are kept constant based on the prior knowledge obtained from trial numerical simulations and experimentation. Efforts have been made to nullify the essence of costly, time consuming experiments towards obtaining high-quality castings out of the findings obtained from numerical simulations. The optimum pouring temperature identified in the present study is 610 °C, which facilitates smoother slurry flow, minimum surface defect concentration, uniform temperature field and solid fraction distribution within the component cavity

Microscale deformation behavior of rheocast Al–7Si–0.3 Mg alloy

In the present work, microscale deformation behavior, plastic strain localization, and plastic instability of rheocast Al–Si–Mg (A356) alloy have been investigated using micromechanical approach. For this purpose, two-dimensional microscale models (representative volume elements) have been developed using actual microstructure of the cast samples made under three different process conditions. Microstructure of the above-mentioned alloy consists of two different phases, such as aluminum-rich primary phase and silicon-rich eutectic phase. In line with that, composite micromechanical models have been developed to analyze them within the finite element framework. Rheocasting has been performed using cooling slope with two different slope angles of 45° and 60°, and comparison has been made with the conventional cast samples of the alloy that has been cast directly from the superheated molten state. Different boundary conditions have been assumed to perform finite element based simulation, using a popular finite element solver ABAQUS, depending upon the position of representative volume elements on the cylindrical tensile specimen. Under uniaxial tensile loading, ductile failure mode is predicted in the form of plastic strain localization due to incompatible deformation between the phases. This indicates inhomogenity of microstructure that determines the damage initiation process within this material, as there is no damage or failure criterion specified during the finite element analysis. Grain size, shape, and orientation of the primary aluminum phase are found to play a vital role on deformation behavior and failure mode of the materials investigated in this study