Development of a Microarray system for the Rapid and Simultaneous Detection of Bacterial and Viral Foodborne Pathogens

Foodborne diseases are increasingly recognized as a significant global public health problem despite major advances and improvements in the quality of food, water, sanitation and hygiene. However, detection and characterization of foodborne pathogens during outbreak scenarios remains a laborious and time-consuming task. The aim of this work was to develop an oligonucleotide microarray for rapid detection and characterization of the most important infectious bacterial (Campylobacter, Salmonella and Yersinia) and viral (Noroviruses) pathogens found in swine and associated pork products. A total of 272 target regions and genes were identified that were specific for pathogen identification and characterization of specific antimicrobial resistance and virulence determinants. We designed multiple probes (up to three) per gene to increase the sensitivity and specificity of the microarray. After BLAST analysis, a total of 562 probes were finally selected to be printed on to glass slides. Appropriate control strains that were previously characterized in our laboratories by PCR were selected to test the developed arrays. Preliminary results indicated that the designed probes were highly specific and sensitive for identification of tested pathogens and known res1stance and virulence genes present in the selected control strains

Characterisation of multiple hindered settling regimes in aggregated mineral suspensions

Aqueous suspensions of magnesium hydroxide are shown to exhibit low ζ-potential behavior and highly complex settling dynamics. Two distinct regimes of hindered settling behavior are observed on either side of a threshold concentration, ϕ*, of 2.38% v/v, which is considerably below the gel point, ϕg, observed at 5.4 ± 1.6% v/v. The low-concentration regime was characterized by a very large Richardson and Zaki exponent of 146, a factor of 10 larger than that of the high-concentration regime. Michaels and Bolger analysis of the low-concentration regime implies settling governed by large, low-density macroaggregates of 138–147 μm diameter and low intraaggregate packing fractions on the order of 0.05, which is in good agreement with in situ particle characterization undertaken using particle vision and measurement (PVM) and focused-beam reflectance measurements (FBRM). The large macroaggregates must undergo some shear densification within the higher-concentration hindered settling regime in order for the suspension to gel at a concentration of 5.4% v/v. Consequently, fluid flow past small, shear-resistant primary agglomerates, observed within the aggregates using scanning electron microscopy and flow particle image analysis, during aggregate densification may represent the limiting step for dewatering within the high-concentration regime

pEPito: a significantly improved non-viral episomal expression vector for mammalian cells

<p>Abstract</p> <p>Background</p> <p>The episomal replication of the prototype vector pEPI-1 depends on a transcription unit starting from the constitutively expressed <it>Cytomegalovirus </it>immediate early promoter (CMV-IEP) and directed into a 2000 bp long <it>matrix attachment region sequence </it>(MARS) derived from the human β-interferon gene. The original pEPI-1 vector contains two mammalian transcription units and a total of 305 CpG islands, which are located predominantly within the vector elements necessary for bacterial propagation and known to be counterproductive for persistent long-term transgene expression.</p> <p>Results</p> <p>Here, we report the development of a novel vector pEPito, which is derived from the pEPI-1 plasmid replicon but has considerably improved efficacy both <it>in vitro </it>and <it>in vivo</it>. The pEPito vector is significantly reduced in size, contains only one transcription unit and 60% less CpG motives in comparison to pEPI-1. It exhibits major advantages compared to the original pEPI-1 plasmid, including higher transgene expression levels and increased colony-forming efficiencies <it>in vitro</it>, as well as more persistent transgene expression profiles <it>in vivo</it>. The performance of pEPito-based vectors was further improved by replacing the CMV-IEP with the <it>human CMV enhancer/human elongation factor 1 alpha promoter </it>(hCMV/EF1P) element that is known to be less affected by epigenetic silencing events.</p> <p>Conclusions</p> <p>The novel vector pEPito can be considered suitable as an improved vector for biotechnological applications <it>in vitro </it>and for non-viral gene delivery <it>in vivo</it>.</p

Compositional attribution of non-provenienced Maya polychrome vessels

Procedures and a few of the results of the Maya ceramic project are discussed from the perspective of non-provenienced vessel attribution ranging from site specific through a more inferential level to the rather hypothetical. The examples presented serve to illustrate the manner in which compositional and stylistic covariation are viewed in an investigation of Maya Ceramic art. The large data base from neutron activation analysis including archaeologically recovered pottery as well as the stylistically and iconographically elaborate vessels requires continued refinement in our methods of statistical analysis along with gaining a greater understanding of the sources of ceramic compositional variation in the Maya area. The mutually beneficial collaboration between science, art, and archaeology are emphasized

Structure and sedimentation characterisation of sheared Mg(OH)2 suspensions flocculated with anionic polymers

In this study, magnesium hydroxide (Mg(OH)2) suspensions were flocculated using two polyacrylamide-poly(acrylic acid) copolymers with charge densities of 30% or 40%. Structural characteristics, including particle size distribution, shape and fractal dimension of the resultant flocs were investigated using complementary techniques; static light scattering, focused beam reflectance measurement, automated optical microscopy and cryogenic scanning electron microscopy. Sedimentation rates were analysed for 2.5 vol.% dispersions at various polymer concentrations and compared to predictions from a fractal modified Richardson-Zaki (FMRZ) settling model. FMRZ model predictions using the 90th percentile (d90) floc sizes produced the most accurate correlations to experimental settling data, as these larger flocs likely dominate settling dynamics by ‘netting’ smaller particles. Overall, the FMRZ settling model provided a first approximation of zonal settling rates, but when further examined by multivariate analysis, was found to be critically sensitive to small changes in fractal dimension

A FENE-P k − ε viscoelastic turbulence model valid up to high drag reduction without friction velocity dependence

A viscoelastic turbulence model in a fully-developed drag reducing channel flow is improved, with turbulent eddies modelled under a k–ε representation, along with polymeric solutions described by the finitely extensible nonlinear elastic-Peterlin (FENE-P) constitutive model. The model performance is evaluated against a wide variety of direct numerical simulation data, described by different combinations of rheological parameters, which is able to predict all drag reduction (low, intermediate and high) regimes with good accuracy. Three main contributions are proposed: one with a simplified viscoelastic closure for the NLTij term (which accounts for the interactions between the fluctuating components of the conformation tensor and the velocity gradient tensor), by removing additional damping functions and reducing complexity compared with previous models; second through a reformulation for the closure of the viscoelastic destruction term, Eτp, which removes all friction velocity dependence; lastly by an improved modified damping function capable of predicting the reduction in the eddy viscosity and thus accurately capturing the turbulent kinetic energy throughout the channel. The main advantage is the capacity to predict all flow fields for low, intermediate and high friction Reynolds numbers, up to high drag reduction without friction velocity dependence

The Contrasting Behavior of Strongly and Weakly Interfacially Active Asphaltenes on the Rheology of Model Waxy Oils

Asphaltenes and waxes are two components of crude oil that cause flow assurance issues. Although the components coexist, few studies have considered the effect of asphaltenes on wax crystallization and gel-forming properties. Furthermore, the current understanding remains contradictory with both wax-alleviating and wax-aggravating behaviors observed. In this study, asphaltenes extracted from a heavy crude oil were fractionated into strongly and weakly interfacially active asphaltenes by partitioning at a water–oil interface. The two asphaltene fractions exhibited contrasting physicochemical properties, with the strongly interfacially active asphaltenes (IAA) being more polar due to their higher heteroatom content (particularly S and O) and forming larger aggregates in the solution compared to the weakly interfacially active asphaltenes (referred to as remaining asphaltenes, RA). The two asphaltene fractions lowered both the wax gelation temperature and wax appearance temperature; however, the effect was comparable. The unit cell lattice structure of the wax particle remained unchanged in the presence of asphaltenes, but the wax particles were found to be smaller with RA compared to IAA. However, the key finding of the study is how the two asphaltene fractions affected the yield strength of the gelled wax. For RA, the yield strength was lowered with an increasing asphaltene concentration, whereas for IAA, the overall effect was to increase the gel yield strength. Because the properties of the wax particles were largely unchanged by the two asphaltene fractions, the result suggests that the asphaltene–asphaltene interaction contributes to the overall yield strength. It was shown that the interaction between RA and RA is repulsive with negligible adhesion, whereas that between IAA and IAA is attractive with strong adhesion. These structure-breaker and structure-maker properties of the two asphaltenes confirm that the asphaltene–asphaltene interaction significantly contributes to modifying the yield strength of a waxy gel

Integron-mediated Multidrug Resistance in a Global Collection of Nontyphoidal Salmonella enterica Isolates

Horizontal gene transfer and clonal expansion contribute substantially to the dissemination of resistant strain

Amphiphilic block copolymers as dual flocculation-flotation agents for rapid solid–liquid separation of radioactive wastes

The potential of poly(acrylic acid)-b-poly(n-butyl acrylate) as a dual flocculant-collector in combined flotation-sedimentation dewatering operations was investigated. The amphiphilic block copolymers were synthesised with consistent hydrophilic chain lengths and varying hydrophobic chain lengths. Various techniques were employed to analyse polymer behaviour at the air–water interface, being interfacial surface tension and dilational viscoelasticity. Polymer adsorption onto Mg(OH)2 was determined differentially using UV–Vis spectroscopy. Floc structures were determined using static light scattering, and flocculation-flotation performance was analysed using settling tests and flotation cell material balances. Results showed that longer hydrophobic chains were less surface-active, reducing foamability and water entrainment. The unimer-micellar adsorption transition points were identified through viscoelastic properties and particle adsorption studies. A distinct change in floc density and structure was observed for the largest molecular weight copolymer when the dosed concentration increased into the micellar adsorption region, suggesting a pseudo-bridging flocculation mechanism. Settling rates were significantly higher for particles flocculated with the larger molecular weight polymer, correlating to their larger aggregate sizes, especially over the micellar transition point. The largest molecular weight block copolymer demonstrated superior collection efficiency compared to the traditional surfactant, sodium dodecylsulfate (SDS), below its micellar adsorption transition point. However, beyond this point, the lack of exposed hydrophobic blocks hindered the hydrophobisation of Mg(OH)2 particles, reducing collection efficiency. Comparing flotation cell particle size distributions, it was suggested that recovery may be hydrodynamically hindered by the largest floc sizes, though recovery was observed for particles in the order of < 600 μm