Increase in bacteraemia cases in the East Midlands region of the UK due to MDREscherichia coliST73: high levels of genomic and plasmid diversity in causative isolates

Objectives: To determine the population structure of E. coli ST73 isolated from human bacteraemia and urinary tract infections. Methods: The genomes of 22 E.coli ST73 isolates were sequenced using the Illumina HiSeq platform. High resolution SNP typing was used to create a phylogenetic tree. Comparative genomics were also performed using a pangenome approach. In silico and S1-PFGE plasmid profiling was conducted, and isolates were checked for their ability to survive exposure to human serum. Results: E.coli ST73 isolates circulating in clinically unrelated episodes show a high degree of diversity at a whole genome level, though exhibit conservation in gene content, particularly in virulence associated gene carriage. The isolates also contain a highly diverse plasmid pool that confers multi-drug resistance via carriage of CTX-M genes. All strains are highly serum resistant and uniformly carry genes shown to be essential for serum resistance. Conclusions: Our data shows that a rise in incidence of multi-drug resistant E.coli ST73 clinical isolates is not due to a circulating outbreak strain as in E.coli ST131. Rather the ST73 circulating strains are distantly related and carry a diverse set of resistance plasmids. This suggests that the evolutionary events behind emergence of drug resistant E.coli differ between lineages

Phenotypic microarrays suggest Escherichia coli ST131 is not a metabolically distinct lineage of extra-intestinal pathogenic E. coli

Extraintestinal pathogenic E. coli (ExPEC) are the major aetiological agent of urinary tract infections (UTIs) in humans. The emergence of the CTX-M producing clone E. coli ST131 represents a major challenge to public health worldwide. A recent study on the metabolic potential of E. coli isolates demonstrated an association between the E. coli ST131 clone and enhanced utilisation of a panel of metabolic substrates. The studies presented here investigated the metabolic potential of ST131 and other major ExPEC ST isolates using 120 API test reagents and found that ST131 isolates demonstrated a lower metabolic activity for 5 of 120 biochemical tests in comparison to non-ST131 ExPEC isolates. Furthermore, comparative phenotypic microarray analysis showed a lack of specific metabolic profile for ST131 isolates countering the suggestion that these bacteria are metabolically fitter and therefore more successful human pathogens

Populations of Escherichia coli in clinical samples of Urinary Tract Infections and bacteraemia

Extraintestinal pathogenic E. coli (ExPEC) strains are the main etiologic agent of urinary tract infections (UTIs). ExPEC strains are also reported to be the most common cause of bacteraemia in the world, which often originate from UTI. The population structure of UTI E. coli strains is well described in the literature with increased prevalence of multidrug resistance driven by extended spectrum β lactamases (ESBLs). ESBL carriage and multidrug resistance of bacteraemia E. coli is on the increase yet little information is available about their population structure. With the aim to define the bacteraemia population structure, E. coli isolated from urine samples and blood cultures were collected from the Nottingham University Hospital NHS trust over a five month period. Isolates were tested for antimicrobial resistance, ESBL and virulence associated gene (VAG) carriage, and were typed by MLST. Significantly higher ESBL driven multidrug resistant strains were observed in the bacteraemia E. coli compared to the UTI isolates with no significant difference in the carriage of VAGs. Our data shows a reduction in population diversity within the bacteraemia isolates compared to the concomitant urine sample population resulting in a small number of dominant sequence types (STs) (ST131, ST73, ST95) which is associated with ESBL conferred multi drug resistance and not specific virulence genes. This suggests that the increased prevalence of ESBL carriage in ExPEC isolates is leading to a selective advantage in a small number of dominant lineages causing bacteraemia in patients. Comparative genome analysis of selected isolates belonging to the dominant ST (ST73) from bacteraemia and UTI was performed to investigate the presence of bacteraemia specific loci that may explain the loss of diversity in bacteraemia. No genomic regions were identified specific for the bacteraemia ST73 isolates other than ESBL carriage. Plasmid profiling of the ESBL positive isolates of this ST73 group from bacteraemia and UTI identified diverse types of plasmids spread between the strains. No specific genomic loci were identified specific for ESBL positive ST73 isolates from bacteraemia and UTI. This concludes that random acquisition of ESBL plasmids by any ST73 E. coli may select for its progression to bacteraemia which is serious and debilitating. Our study provided a comprehensive snapshot of the E.coli population structure from contemporaneous clinical cases of UTI and bacteraemia. The large increase in multi-drug resistance in bacteraemia ExPEC populations compared to co-circulating UTI populations is of clinical concern and represents a challenge in control and treatment of serious extra-intestinal E. coli infections. This provides an important clinical insight into how common E. coli STs could adapt to become dominant bacteraemia agents

Combined Solutal and Thermal Buoyancy Thermocapillary Convection in a Square Open Cavity

Combined solutal and thermal buoyancy–thermocapillary convection in a square open cavity is studied numerically in the present article. The Forchheimer–Brinkman-extended Darcy model is used in the mathematical formulation for the porous layer and the COMSOL Multiphysics software is applied to solve the dimensionless governing equations. The governing parameters considered are the thermal Marangoni number, −1000 ≤ Ma_T ≤ 1000, the Darcy number, 10−5 ≤ Da ≤ 10−2, the porosity of porous medium, 0.4 ≤ ε ≤ 0.99 and the Lewis number, 10 ≤ Le ≤ 200. It is found that the global heat and solute transfer rate decreases by reducing the counteracting surface tension force and increases by augmenting the surface tension force. The minimum values of the global heat and solute transfer rate were obtained about Ma_T = −90 for the all porosities

Serum albumin and osmolality inhibit Bdellovibrio bacteriovorus predation in human serum

We evaluated the bactericidal activity of Bdellovibrio bacteriovorus, strain HD100, within blood sera against bacterial strains commonly associated with bacteremic infections, including E. coli, Klebsiella pneumoniae and Salmonella enterica. Tests show that B. bacteriovorus HD100 is not susceptible to serum complement or its bactericidal activity. After a two hour exposure to human sera, the prey populations decreased 15- to 7,300-fold due to the serum complement activity while, in contrast, the B. bacteriovorus HD100 population showed a loss of only 33%. Dot blot analyses showed that this is not due to the absence of antibodies against this predator. Predation in human serum was inhibited, though, by both the osmolality and serum albumin. The activity of B. bacteriovorus HD100 showed a sharp transition between 200 and 250 mOsm/kg, and was progressively reduced as the osmolality increased. Serum albumin also acted to inhibit predation by binding to and coating the predatory cells. This was confirmed via dot blot analyses and confocal microscopy. The results from both the osmolality and serum albumin tests were incorporated into a numerical model describing bacterial predation of pathogens. In conclusion, both of these factors inhibit predation and, as such, they limit its effectiveness against pathogenic prey located within sera

Broadening the antibacterial spectrum of histidine kinase autophosphorylation inhibitors via the use of epsilon-poly-L-lysine capped mesoporous silica-based nanoparticles

[EN] Two-component systems (TCS) regulate diverse processes such as virulence, stress responses, metabolism and antibiotic resistance in bacteria but are absent in humans, making them promising targets for novel antibacterials. By incorporating recently described TCS histidine kinase autophosphorylation inhibitors (HKAIs) into epsilon-poly-L-lysine capped nanoparticles (NPs) we could overcome the Gram negative (Gr(-)) permeability barrier for the HKAIs. The observed bactericidal activity against Gr(-) bacteria was shown to be due to the enhanced delivery and internalization of the HKAIs and not an inhibitory or synergistic effect of the NPs. The NPs had no adverse effects on mammalian cell viability or the immune function of macrophages in vitro and showed no signs of toxicity to zebrafish larvae in vivo. These results show that HKAIs are promising antibacterials for both Gr(-) and Gr + pathogens and that NPs are a safe drug delivery technology that can enhance the selectivity and efficacy of HKAIs against bacteria. (C) 2016 Elsevier Inc. All rights reserved.This work was funded by FP7 ITN STARS-Scientific Training in Antimicrobial Research Strategies (Contract No. PITN-GA-2009-238490, J.M.W., A.M.), H2020 MSCA IF (AND-659121, N.V.), grant BIO2013-42619-P from the Ministerio de Economia y Competitividad (A.M.), grant from the Spanish Government (Project MAT2015-64139-C4-1-R,N. M., J.R.M, R.M.M.), and a grant from Generalitat Valenciana (Project PROMETEOII/2014/047, N.M.). and Prometeo II/2014/029, A.M.).Velikova, N.; Mas Font, N.; Miguel-Romero, L.; Polo, L.; Stolte, E.; Zaccaria, E.; Cao, R.... (2017). Broadening the antibacterial spectrum of histidine kinase autophosphorylation inhibitors via the use of epsilon-poly-L-lysine capped mesoporous silica-based nanoparticles. Nanomedicine Nanotechnology Biology and Medicine. 13(2):569-581. https://doi.org/10.1016/j.nano.2016.09.011S56958113

Thermal-Diffusive Processes of an Electron-Hole Non-Local Semiconductor Model with Variable Thermal Conductivity and Hall Current Effect

In this work, a novel model is presented that describes thermal diffusion processes through non-local semiconductor materials. The material under study is subjected to the influence of a strong magnetic field, which creates a Hall current. Interference between the excited electrons and the excited holes of a non-local semiconductor that had been exposed to temperature was present, and thermal conductivity depending on changes in graduated temperature were accounted for. The governing equations are written in a dimensionless form in one dimension (1D) where the thermal conductivity is taken as a function of temperature through electronic and elastic deformation (ED and ED) processes. Laplace transforms in one dimension with initial conditions were used to convert partial differential equations to arrive at exact formulas of solutions. To obtain the exact linear solutions, some boundary conditions taken on the free surface of the non-local semiconductor were used. Using numerical methods of inverse Laplace transforms, the complete solutions of the physical quantities under study were obtained. To further understand how various variables (thermal memory, variable thermal conductivity, and Hall current) affect the non-local semiconductor, numerical physical fields were simulated, and are graphically depicted, and discussed herein

Free convection from a corrugated heated cylinder with nanofluids in a porous enclosure

Natural convection between a cold square porous enclosure and a hot corrugated cylinder is studied numerically in the current article. The enclosure is filled with a water-base nanofluids suspending metal nanoparticles and the porous layer is modelled applying the Brinkman-Forchheimer law. The finite element method has been utilised to solve the governing equations. Analysis in this studies are: the amplitude of corrugated surface, the number of corrugated surface and the concentration are considered. It is found that the heat transfer of the corrugated cylinder might be slightly better than the heat transfer of the smooth cylinder under specific circumstances, but in general, the heat transfer is reduced by applying the corrugated surface. The heat transfer enhances up to [Formula: see text] by increasing nanoparticle concentration. The heat transfer rate does not increase linearly by increasing the concentration, but it is proportional to the square root of the concentration. </jats:p

Natural convection of Nanoliquid from a Cylinder in Square Porous Enclosure using Buongiorno’s Two-phase Model

AbstractNatural convection of nanoliquid in a square porous enclosure has been studied using non homogeneous two-phase Buongiorno’s model. The outer of enclosure has cold temperature and a circular cylinder is put at the center. A finite heated segment is located on the top cylinder surface which is otherwise insulated. The momentum in the porous layer is modeled applying the Brinkman-Forchheimer equations. The analysis are conducted in the following interval of the associated groups: the portion of heated surface (5% ≤ H ≤ 100%), the concentration (0.0 ≤ ϕ ≤ 0.04), the Darcy number, 10−5 ≤ Da ≤ 10−2 and the cylinder size, (0.15 ≤ R ≤ 0.25). The minimum heat transfer rate of the active surface were obtained at location ξ = 90°. In general, the ratio of the heat transfer per unit area of the heat source decreases as the length of the heated surface increases. The heat transfer rate is intensified for the half thermally active surface and high value of Darcy number at higher nanoparticles concentration.</jats:p

Free Convection Heat Transfer in Composite Enclosures with Porous and Nanofluid Layers

This work conducts a numerical investigation of convection heat transfer within two composite enclosures. These enclosures consist of porous and nanofluidic layers, where the porous layers are saturated with the same nanofluid. The first enclosure has two porous layers of different sizes and permeabilities, while the second is separated by a single porous layer. As the porous layer thickness approaches zero, both enclosures transition to clear nanofluid enclosures. The study uses the Navier–Stokes equations to govern fluid flow in the nanofluid domain and the Brinkman–Forchheimer extended Darcy model to describe flow within the saturated porous layer. Numerical solutions are obtained using an iterative finite difference method. Key parameters studied include the porous thickness (0.0≤S≤1.0), the nanoparticle volume fraction (0.0≤ϕ≤0.05), the thermal conductivity ratio (0.5≤Rk≤10), and the Darcy number (10−5≤Da≤10−2). Key findings include the observation that the highest heat transfer is achieved at the highest concentration, regardless of the porous layer configuration, permeability value, or thermal conductivity ratio. Specifically, an augmentation in values of Nu―I up to 22% is obtained as concentration is adjusted from 1% to 5%. Similarly, an augmentation in values of Nu―II up to 25% is obtained as concentration is adjusted from 1% to 5%