Metal-Pseudomonas veronii 2E Interactions as Strategies for Innovative Process Developments in Environmental Biotechnology.

The increase of industrial discharges is the first cause of the contamination of water bodies. The bacterial survival strategies contribute to the equilibrium restoration of ecosystems being useful tools for the development of innovative environmental biotechnologies. The aim of this work was to study the Cu(II) and Cd(II) biosensing, removal and recovery, mediated by whole cells, exopolymeric substances (EPS) and biosurfactants of the indigenous and non-pathogenic Pseudomonas veronii 2E to be applied in the development of wastewater biotreatments. An electrochemical biosensor was developed using P. veronii 2E biosorption mechanism mediated by the cell surface associated to bound exopolymeric substances. A Carbon Paste Electrode modified with P. veronii 2E (CPEM) was built using mineral oil, pre-washed graphite power and 24 h-dried cells. For Cd(II) quantification the CPEM was immersed in Cd(II) (1-25 µM), detected by Square Wave Voltammetry. A similar procedure was used for 1-50 µM Cu(II). Regarding Cd(II), removal mediated by immobilized EPS was tested in a 50 ml bioreactor with 0.13 mM Cd(II), pH 7.5. A 54% metal retention by EPS was achieved after 7 h of continuous operation, while a 40% was removed by a control resin. In addition, surfactants produced by P. veronii 2E were studied for recovery of Cd(II) adsorbed on diatomite, obtaining a 36% desorption efficiency at pH 6.5. Cu(II) adsorption from a 1 mM solution was tested using P. veronii 2E purified soluble EPS in 50 mL- batch reactors (pH = 5.5, 32°C). An 80% of the initial Cu(II) was retained using 1.04 g immobilized EPS. Focusing on metal recovery, Cu nanoparticles (NPs) biosynthesis by P. veronii 2E was carried out in Cu(II)-PYG Broth at 25°C for 5 days. Extracellular CuNPs were characterized by UV-Vis spectral analysis while both extracellular and intracellular NPs were analyzed by SEM and TEM techniques. Responses of P. veronii 2E and its products as biosurfactants, bound and soluble EPS allowed Cu(II) and Cd(II) removal, recovery and biosensing resulting in a multiple and versatile tool for sustainable wastewater biotreatments.

Structural characterization and metal biosorptive activity of the major polysaccharide produced by Pseudomonas veronii 2E.

Pseudomonas veronii 2E, an autochthonous bacterium isolated from sediments associated to a high-polluted watershed, produces a complex matrix of exopolymers with carbohydrates as main components. In this work, four polysaccharides were isolated from the extracellular material. The major acidic polysaccharide named EPO2, was purified and its structure was elucidated using Matrix-assisted laser desorption/ionization and Electrospray ionization mass spectrometry, Infrared spectroscopy, Nuclear magnetic resonance spectroscopy and chemical treatments. This heteropolysaccharide consists in an α(1-4) glucan substituted with N-Acetylglucosamine residues and with a branching α-D-GlcpA-(1-3)-L-Fucp disaccharide. The biosorption capacity of EPO2 and of the whole exopolysaccharide to Pb(II), Zn(II), Cu(II) and Fe(II) was evaluated. EPO2 showed a remarkable sorption capacity for Fe(II) with an efficiency of 70% and for Zn(II) 39%. When the whole exopolysaccharide fraction was tested it showed a significantly lower metal sorption ability than purified EPO2 suggesting the involvement of the distinct acidic branching disaccharide in this interaction.

Online self-powered Cr(VI) monitoring with autochthonous Pseudomonas and a bio-inspired redox polymer.

The assessment of water quality is critical to implement preventive and emergency interventions aimed to limit/avoid environmental contamination and human exposure to toxic compounds. While established high-resolution techniques allow quantitative and qualitative determination of contaminants, their widespread application is not feasible due to cost, time, and need for trained personnel. In this context, the development of easy-to-implement approaches for preliminary detection of contaminants is of the utmost importance. Herein, a portable self-powered microbial electrochemical sensor enabling online monitoring of Cr(VI) is reported. The biosensor employs a bio-inspired redox mediating system to allow extracellular electron transfer between a bacterial isolate from chromium-contaminated environments and the electrode, providing a clear response to Cr(VI) presence. The biosensor shows good linearity (R2 = 0.983) and a limit of detection of 2.4 mg L-1 Cr(VI), with a sensitivity of 0.31 ± 0.02 µA cm-2 mgCr(VI)-1 L. The presented microbial bioanode architecture enhanced biosensor performance thanks to the improved "electrical wiring" between biological entities and the abiotic electrode surface. This approach could be easily implemented in engineered electrode surfaces, such as paper-based multi-anodes that maximize bacterial colonization, further improving biosensor response. Graphical abstract.

Infrared spectroscopy with multivariate analysis to interrogate the interaction of whole cells and secreted soluble exopolimeric substances of Pseudomonas veronii 2E with Cd(II), Cu(II) and Zn(II).

Extracellular polymeric substances (EPS) are bacterial products associated to cell wall or secreted to the liquid media that form the framework of microbial mats. These EPS contain functional groups as carboxyl, amino, hydroxyl, phosphate and sulfhydryl, able to interact with cations. Thus, EPS may be considered natural detoxifying compounds of metal polluted waters and wastewaters. In this work Attenuated Total Reflectance-Fourier Transform Infrared spectroscopy (ATR-FTIR) in combination with multivariate analysis (Principal Component Analysis-PCA-) were used to study the interaction of Cd(II), Cu(II) and Zn(II) and Pseudomonas veronii 2E cells, including bound EPS and cell wall, and its different soluble EPS fractions, previously characterized as Cd(II) ligands of moderate strength. Amino groups present in exopolysaccharide fraction were responsible for Zn(II) and Cu(II) complexation, while carboxylates chelated Cd(II). In lipopolysaccharide fraction, phosphoryl and carboxyl sites were involved in Cd(II) and Cu(II) binding, while Zn(II) interacted with amino groups. Similar results were obtained from cells. These studies confirmed that FTIR-PCA is a rapid analytical tool to provide valuable information regarding the functional groups in biomolecules related to metal interaction. Moreover, a discrimination and identification of functional groups present in both EPS and cells that interacted with Cd(II), Zn(II) and Cu(II) was demonstrated.

Chemical characterization and ligand behaviour of Pseudomonas veronii 2E siderophores.

Siderophores are low-molecular weight ligands secreted by bacteria as a survival strategy in Fe(III)-lacking environments. They bind not only Fe(III), but Co(II), Zn(II), Mn(II), Ni(II), Ga(III) as a detoxification alternative. The synthesis, purification and characterization of siderophores produced by Pseudomonas veronii 2E were evaluated to be applied in future environmental technologies. Optimal production was obtained in Fe(III)-free M9-succinate at 25 °C, 40 h and pH 6.9. Siderophores were chemically characterized as hydroxamate and catechol mixed-type. Spectroscopic analysis indicated their belonging to the pyoverdine family, behaving as ligand to Cd(II), Zn(II), Cu(II), Ni(II) and Cr(III), which promoted siderophoregenesis during growth. Siderophore-Cd(II) complexation was studied by electrochemical monitored titration revealing one family of moderate-strength binding sites. Mass spectral analysis evidenced the secretion of a variety of molecules (molecular mass ca.1200 u). Non pathogenic Pseudomonas veronii 2E siderophores represent a safe alternative for the concrete application of environmental technologies and clinical procedures.

Characterization and electrochemical response of DNA functionalized 2nm gold nanoparticles confined in a nanochannel array.

Polyvalent gold nanoparticle oligonucleotide conjugates are subject of intense research. Even though 2nm diameter AuNPs have been previously modified with DNA, little is known about their structure and electrochemical behavior. In this work, we examine the influence of different surface modification strategies on the interplay between the meso-organization and the molecular recognition properties of a 27-mer DNA strand. This DNA strand is functionalized with different sulfur-containing moieties and immobilized on 2nm gold nanoparticles confined on a nanoporous alumina, working the whole system as an electrode array. Surface coverages were determined by EXAFS and the performance as recognition elements for impedance-based sensors is evaluated. Our results prove that low DNA coverages on the confined nanoparticles prompt to a more sensitive response, showing the relevance in avoiding the DNA strand overcrowding. The system was able to determine a concentration as low as 100pM of the complementary strand, thus introducing the foundations for the construction of label-free genosensors at the nanometer scale.

Confined gold nanoparticles enhance the detection of small molecules in label-free impedance aptasensors.

A controlled architecture of nanoelectrodes, of a similar size to small molecule-binding aptamers, is synthesized inside nanoporous alumina. Gold nanoparticles with a controlled size (about 2 nm) are electrogenerated in the alumina cavities, showing a fast electron transfer process toward ferrocyanide. These uncapped nanoparticles are easily modified with a thiol-containing aptamer for label-free detection of adenosine monophosphate by electrochemical impedance spectroscopy. Our results show that the use of a limited electrical conducting surface inside an insulating environment can be very sensitive to conformational changes, introducing a new approach to the detection of small molecules, exemplified here by the direct and selective detection of adenosine monophosphate at the nanomolar scale.

Uso de lacosamida en pacientes pediátricos con epilepsia refractaria como terapia adjunta, revisión sistemática de la literatura / Use of Lacosamide in pediatric population with refractory epilepsy as an adjunctive therapy, a systematic review of literature

Objetivo: presentar la evidencia publicada hasta el momento acerca de la utilidad de la lacosamida en población pediátrica con epilepsia refractaria. Métodos: se realizaron búsquedas en las bases de datos electrónicas PubMed, Embase, Cochrane y Scielo, encontrando un total de 18 artículos que cumplieron los criterios de búsqueda. Hallazgos principales : lacosamida, uno de los fármacos anticonvulsivantes nuevos, ha sido aprobado como terapia adjunta en epilepsias de inicio focal y epilepsias refractarias en pacientes mayores de 16 años, pero existe creciente evidencia de su eficacia y seguridad en población pediátrica desde los 4 años. Los efectos adversos no difieren en gran medida de los demás anticonvulsivantes y su tasa de éxito es de aproximadamente el 8-40% de control total de crisis. Sin embargo, los estudios no tienen un seguimiento a largo plazo que demuestre que se mantiene este mismo control de crisis en el transcurso del tiempo. Adicionalmente, la lacosamida ofrece mejoría clínica independientemente del control de las crisis en cuanto al patrón motor y comportamental. Conclusiones: Los estudios muestran que la lacosamida es una opción segura indicada para la epilepsia de inicio focal y epilepsia refractaria en la población pediátrica como terapia adjunta, debido a su efectividad en el control de crisis, perfil de seguridad, baja tasa de efectos adversos e interacción con otros medicamentos; no obstante, se requieren estudios con mayor número de pacientes y con seguimientos más prolongados.

Objetive: to present the evidence published about the usefulness of lacosamide in pediatric patients with refractory epilepsy, in order to explain its use in this population. Methods: we conducted search in PubMed, Embase, Cochrane library and Scielo and found 18 articles that fulfilled the inclusion criteria. Findings: lacosamide, is one of the newer AED (Antiepileptic Drugs) approved for focal refractory and focal onset epilepsies as an add on treatment in patients older than 16 years. However there is growing evidence of its use, security and efficacy in pediatric population from 4 years and above. Adverse effects do not differ from others found in others AED with success seizure free rates between 8 and 40%. Studies revisited had major issues on long term following. In addition, lacosamide improves behavioral and motor skills in children despite seizure control. Conclusions: Studies shows lacosamide as a safe option in pediatric patients with focal onset and refractory epilepsies as an add on therapy given its seizure control, safety profile, low rate of adverse effects and interaction between others medications, however larger and long term studies are needed.

Cadmium, zinc and copper biosorption mediated by Pseudomonas veronii 2E.

Adsorption properties of bacterial biomass were tested for Cd removal from liquid effluents. Experimental conditions (pH, time, cellular mass, volume, metal concentration) were studied to develop an efficient biosorption process with free or immobilised cells of Pseudomonas veronii 2E. Surface fixation was chosen to immobilise cells on inert surfaces including teflon membranes, silicone rubber and polyurethane foam. Biosorption experiments were carried out at 32 degrees C and controlled pH; maximal Cd(II) retention was observed at pH 7.5. The isotherm followed the Langmuir model (K(d)=0.17 mM and q(max)=0.48 mmol/g cell dry weight). Small changes in the surface negative charge of cells were observed by electrophoretic mobility experiments in presence of Cd(II). In addition, biosorption of 40% Cu(II) (pH 5 and 6.2) and 50% Zn(II) and 50% Cd(II) (pH 7.5) was observed from mixtures of Cu(II), Zn(II) and Cd(II) 0.5mM each.

Indigenous heavy metal multiresistant microbiota of Las Catonas stream.

Las Catonas stream (Buenos Aires Metropolitan Area) receives a complex mixture of pollutants from point and diffuse sources because of the agricultural, industrial and urban land uses of its basin. Widespread detection of heavy metals exceeding aquatic life protection levels has occurred in monitoring reconnaissance studies in surface and pore water. As a result of the screening of Cu, Cd, Zn and Pb resistant/tolerant and culturable microbiota, B101N and 200H strains (Pseudomonas fluorescens or putida) were isolated and selected for further studies. They showed 65% Cd and 35% Zn extraction efficiency from aqueous phase. The potential use of these strains in wastewater treatment is currently investigated in order to contribute to decrease heavy metal pollution, a problem affecting every stream of Buenos Aires Metropolitan Area.

Descripción del estado de las aguas de los arroyos Claro y Las Tunas en el triángulo industrial de Malvinas Argentina

Se presenta un estudio fisico-quimico del estado basal de los arroyos Claro y Las Tunas, ubicados en la Region Metropolitana Buenos Aires (RMBA). Ambos arroyos son de bajo caudal y poseen lenta velocidad, determinada por una topografia de pendiente suave. Los tramos superiores atraviesan zonas de media a alta densidad de poblacion, internandose luego en regiones donde se concentra una intensa actividad industrial

Descripción del estado de las aguas de los arroyos Claro y Las Tunas en el triángulo industrial de Malvinas Argentina

Se presenta un estudio fisico-quimico del estado basal de los arroyos Claro y Las Tunas, ubicados en la Region Metropolitana Buenos Aires (RMBA). Ambos arroyos son de bajo caudal y poseen lenta velocidad, determinada por una topografia de pendiente suave. Los tramos superiores atraviesan zonas de media a alta densidad de poblacion, internandose luego en regiones donde se concentra una intensa actividad industrial