Whole genome sequencing,molecular typing and in vivovirulence of OXA-48-producingEscherichia coli isolates includingST131 H30-Rx, H22 and H41subclones

Carbapenem-resistant Enterobacteriaceae, including the increasingly reported OXA-48 Escherichia coli producers, are an emerging public health threat worldwide. Due to their alarming detection in our healthcare setting and their possible presence in the community, seven OXA-48-producing, extraintestinal pathogenic E. coli were analysed by whole genome sequencing as well as conventional tools, and tested for in vivo virulence. As a result, five E. coli OXA-48-producing subclones were detected (O25:H4-ST131/PST43-fimH30-virotype E; O25:H4-ST131/PST9-fimH22-virotype D5, O16:H5-ST131/ PST506-fimH41; O25:H5-ST83/PST207 and O9:H25-ST58/PST24). Four ST131 and one ST83 isolates satisfied the ExPEC status, and all except the O16:H5 ST131 isolate were UPEC. All isolates exhibited local inflammatory response with extensive subcutaneous necrosis but low lethality when tested in a mouse sepsis model. The blaOXA-48 gene was located in MOBP131/IncL plasmids (four isolates) or within the chromosome (three ST131 H30-Rx isolates), carried by Tn1999-like elements. All, except the ST83 isolate, were multidrug-resistant, with additional plasmids acting as vehicles for the spread of various resistance genes. This is the first study to analyse the whole genome sequences of blaOXA-48-positive ST131, ST58 and ST83 E. coli isolates in conjunction with experimental data, and to evaluate the in vivo virulence of blaOXA-48 isolates, which pose an important challenge to patient management

Antioxidant and cytotoxic activities of sulfated polysaccharides from five different edible seaweeds

In recent times, there has been a growing interest in the exploration of antioxidants and global trend toward the usage of seaweeds in the food industries. The low molecular weight up to 14 kDa sulfated polysaccharides of seaweeds (Portieria hornemannii, Spyridia hypnoides, Asparagopsis taxiformis, Centroceras clavulatum and Padina pavonica) were evaluated for in vitro antioxidant activities and cytotoxic assay using HeLa cell line and also characterized by FTIR. The high yield (7.74% alga dry wt.) of sulfated polysaccharide was observed in P. hornemannii followed by S. hypnoides (0.69%), C. clavulaum (0.55%) and A. taxiformis (0.17%). In the brown seaweed P. pavonica, the sulfated polysaccharide yield was 2.07%. High amount of sulfate was recorded in the polysaccharide of A. taxiformis followed by C. clavulaum, P. pavonica, S. hypnoides and P. hornemannii as indicative for bioactivity. The FTIR spectroscopic analysis supports the sulfated polysaccharides of S. hypnoides, C. clavulatum and A. taxiformis are similar to agar polymer whereas the spectral characteristics of P. hornemannii have similarities to carrageenan. The higher DPPH activity and reducing power were recorded in the polysaccharide of brown seaweed P. pavonica than the red seaweeds as follows: DPPH activities: S. hypnoides > A. taxiformis > C. clavulatum > P. hornimanii; Reducing power: A. taxiformis > P. hornimanii > S. hypnoides > C. clavulatum. The polysaccharide fractions contain up to 14 kDa from red seaweeds P. hornemannii and S. hypnoides followed by brown seaweed P. pavonica exhibit cytotoxic activity in HeLa cancer cell line (and are similar to structural properties of carrageenan extracted from P. hornemannii). The low molecular weight agar like polymer of S. hypnoides and alginate like brown seaweed P. pavonica showing better in vitro antioxidant activities that are capable of exhibiting cytotoxicity against HeLa cell line can be taken up further in-depth investigation for nutraceutical study.University of Algarve: DL 57/2016info:eu-repo/semantics/publishedVersio

Functionalized Palladium Nanoparticles for Hydrogen Peroxide Biosensor

We present a comparison between two biosensors for hydrogen peroxide (H2O2) detection. The first biosensor was developed by the immobilization of Horseradish Peroxidase (HRP) enzyme on thiol-modified gold electrode. The second biosensor was developed by the immobilization of cysteamine functionalizing palladium nanoparticles on modified gold surface. The amino groups can be activated with glutaraldehyde for horseradish peroxidase immobilization. The detection of hydrogen peroxide was successfully observed in PBS for both biosensors using the cyclic voltammetry and the chronoamperometry techniques. The results show that the limit detection depends on the large surface-to-volume ratio attained with palladium nanoparticles. The second biosensor presents a better detection limit of 7.5 μM in comparison with the first one which is equal to 75 μM

Mass transfer characterization in Donnan dialysis

International audienc

Calibration of Surface Plasmon Resonance Imager for Biochemical Detection

We present a new Surface Plasmon Resonance imager (SPRi) based on immobilized T4-phage for bacteria detection. First, we present the sensitivity of the SPR imager towards refractive index variation for biosensor application. The SPR imager can be calibrated versus different percentage of triethylene glycol mixture in ultrapure water. The system can be used as a refractometer with sensitivity below 5 × 10 −5 in the range of 1.33300-1.34360. Second, bacteriophage (T4-phage) can be physisorbed on gold microarray spots for bacteria detection. The kinetic physisorption of different concentrations of T4-phages can be observed in real time. Finally, two types of bacteria such as E. coli (gram negative) and Lactobacillus (gram positive) were used for positive and negative tests. The results show a selectivity of T4-phage toward E. coli with a detection limit below 10 4 CFU/mL and with good reproducibility

Impedance biosensing using phages for bacteria detection: Generation of dual signals as the clue for in-chip assay confirmation

In the present work, we compare the use of antibodies (Ab) and phages as bioreceptors for bacteria biosensing by Electrochemical Impedance Spectroscopy (EIS). With this aim, both biocomponents have been immobilised in parallel onto interdigitated gold microelectrodes. The produced surfaces have been characterised by EIS and Fourier Transform Infra-Red (FTIR) Spectroscopy and have been applied to bacteria detection. Compared to immunocapture, detection using phages generates successive dual signals of opposite trend over time, which consist of an initial increase in impedance caused by bacteria capture followed by impedance decrease attributed to phage-induced lysis. Such dual signals can be easily distinguished from those caused by non-specific adsorption and/or crossbinding, which helps to circumvent one of the main drawbacks of reagentless biosensors based in a single target-binding event. The described strategy has generated specific detection of E. coli in the range of 1E4 - 1E7 CFU mL-1 and minimal interference by non-target Lactobacillus. We propose that the utilization of phages as capture biocomponent for bacteria capture and EIS detection allows in-chip signal confirmation.Peer reviewe