Vert-le-Petit, France
Vert-le-Petit, France

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Al Dahouk S.,German Federal Institute for Risk Assessment | Al Dahouk S.,RWTH Aachen | Hofer E.,Austrian Agency for Health and Food Safety | Tomaso H.,Friedrich Loeffler Institute | And 11 more authors.
Applied and Environmental Microbiology | Year: 2012

Brucellosis is one of the major bacterial zoonoses worldwide. In the past decade, an increasing number of atypical Brucella strains and species have been described. Brucella microti in particular has attracted attention, because this species not only infects mammalian hosts but also persists in soil. An environmental reservoir may pose a new public health risk, leading to the reemergence of brucellosis. In a polyphasic approach, comprising conventional microbiological techniques and extensive biochemical and molecular techniques, all currently available Brucella microti strains were characterized. While differing in their natural habitats and host preferences, B. microti isolates were found to possess identical 16S rRNA, recA, omp2a, and omp2b gene sequences and identical multilocus sequence analysis (MLSA) profiles at 21 different genomic loci. Only highly variable microsatellite markers of multiple-locus variable-number tandem repeat (VNTR) analysis comprising 16 loci (MLVA-16) showed intraspecies discriminatory power. In contrast, biotyping demonstrated striking differences within the genetically homologous species. The majority of the mammalian isolates agglutinated only with monospecific anti-M serum, whereas soil isolates agglutinated with anti-A, anti-M, and anti-R sera. Bacteria isolated from animal sources were lysed by phages F1, F25, Tb, BK2, Iz, and Wb, whereas soil isolates usually were not. Rough strains of environmental origin were lysed only by phage R/C. B. microti exhibited high metabolic activities similar to those of closely related soil organisms, such as Ochrobactrum spp. Each strain was tested with 93 different substrates and showed an individual metabolic profile. In summary, the adaptation of Brucella microti to a specific habitat or host seems to be a matter of gene regulation rather than a matter of gene configuration. © 2012, American Society for Microbiology.


Dorandeu F.,Institute Of Recherche Biomedicale Des Armees | Taysse L.,Center dEtudes du Bouchet | Boudry I.,Institute Of Recherche Biomedicale Des Armees | Foquin A.,Institute Of Recherche Biomedicale Des Armees | And 5 more authors.
Human and Experimental Toxicology | Year: 2011

Exposure to lethal chemical warfare agents (CWAs) is no longer only a military issue due to the terrorist threat. Among the CWAs of concern are the organophosphorus nerve agent O-ethyl-S-(2[di-isopropylamino]ethyl)methyl- phosphonothioate (VX) and the vesicant sulfur mustard (SM). Although efficient means of decontamination are available, most of them lose their efficacy when decontamination is delayed after exposure of the bare skin. Alternatively, CWA skin penetration can be prevented by topical skin protectants. Active research in skin protection and decontamination is thus paramount. In vivo screening of decontaminants or skin protectants is usually time consuming and may be expensive depending on the animal species used. We were thus looking for a suitable, scientifically sound and cost-effective model, which is easy to handle. The euthymic hairless mouse Crl: SKH-1 (hr/hr) BR is widely used in some skin studies and has previously been described to be suitable for some experiments involving SM or SM analogs. To evaluate the response of this species, we studied the consequences of exposing male anaesthetized SKH-1 mice to either liquid VX or to SM, the latter being used in liquid form or as saturated vapours. Long-term effects of SM burn were also evaluated. The model was then used in the companion paper (Taysse et al.1). © The Author(s) 2010.


Taysse L.,Center dEtudes du Bouchet | Dorandeu F.,Institute Of Recherche Biomedicale Des Armees | Daulon S.,Center dEtudes du Bouchet | Foquin A.,Institute Of Recherche Biomedicale Des Armees | And 3 more authors.
Human and Experimental Toxicology | Year: 2011

Using the hairless mouse screening model presented in the companion paper1 the aim of this study was to assess two skin decontaminating systems: Fuller's earth (FE) and Reactive Skin Decontamination Lotion (RSDL) against two extremely toxic chemical warfare agents that represent a special percutaneous hazard, sulphur mustard (SM) and O-ethyl-S-(2[di-isopropylamino] ethyl)methyl-phosphonothioate (VX). Five minutes after being exposed on the back to either 2 μL of neat sulphur mustard or 50 μg.kg-1 of diluted VX, mice were decontaminated. Both systems were able to reduce blisters 3 days after SM exposure. However, RSDL was found to be more efficient than FE in reducing the necrosis of the epidermis and erosion. In the case of VX exposure, RSDL, whatever the ratio of decontaminant to toxicant used (RSDL 10, 20, 50), was not able to sufficiently prevent the inhibition of plasma cholinesterases taken as a surrogate marker of exposure and toxicity. Only FE reduced significantly the ChE inhibition. Some of these observations are different from our previous results obtained in domestic swine and these changes are thus discussed in the perspective of using SKH-1 hairless mice for the initial in vivo screening of decontaminants. © The Author(s) 2010.


Irenge L.M.,Catholic University of Leuven | Durant J.-F.,Catholic University of Leuven | Tomaso H.,University of Federal Defense Munich | Tomaso H.,Institute for Bacterial Infections and Zoonoses | And 5 more authors.
Applied Microbiology and Biotechnology | Year: 2010

A real-time polymerase chain reaction (PCR) assay was developed for rapid identification of Bacillus anthracis in environmental samples. These samples often harbor Bacillus cereus bacteria closely related to B. anthracis, which may hinder its specific identification by resulting in false positive signals. The assay consists of two duplex real-time PCR: the first PCR allows amplification of a sequence specific of the B. cereus group (B. anthracis, B. cereus, Bacillus thuringiensis, Bacillus weihenstephanensis, Bacillus pseudomycoides, and Bacillus mycoides) within the phosphoenolpyruvate/sugar phosphotransferase system I gene and a B. anthracis specific single nucleotide polymorphism within the adenylosuccinate synthetase gene. The second real-time PCR assay targets the lethal factor gene from virulence plasmid pXO1 and the capsule synthesis gene from virulence plasmid pXO2. Specificity of the assay is enhanced by the use of minor groove binding probes and/or locked nucleic acids probes. The assay was validated on 304 bacterial strains including 37 B. anthracis, 67 B. cereus group, 54 strains of non-cereus group Bacillus, and 146 Gram-positive and Gram-negative bacteria strains. The assay was performed on various environmental samples spiked with B. anthracis or B. cereus spores. The assay allowed an accurate identification of B. anthracis in environmental samples. This study provides a rapid and reliable method for improving rapid identification of B. anthracis in field operational conditions. © 2010 Springer-Verlag.


Sharan M.,Indian Institute of Technology Delhi | Singh S.K.,Indian Institute of Technology Delhi | Issartel J.P.,Center dEtudes du Bouchet
Pure and Applied Geophysics | Year: 2012

The identification of single and multiple-point emission sources from limited number of atmospheric concentration measurements is addressed using least square data assimilation technique. During the process, a new two-step algorithm is proposed for optimization, free from initialization and filtering singular regions in a natural way. Source intensities are expressed in terms of their locations reducing the degree of freedom of unknowns to be estimated. In addition, a strategy is suggested for reducing the computational time associated with the multiple-point source identification. The methodology is evaluated with the synthetic, pseudo-real and noisy set of measurements for two and three simultaneous point emissions. With the synthetic data, algorithm estimates the source parameters exactly same as the prescribed in all the cases. With the pseudo-real data, two and three point release locations are retrieved with an average error of 17 m and intensities are estimated on an average within a factor of 2. Finally, the advantages and limitations of the proposed methodology are discussed. © 2011 Springer Basel AG.


Issartel J.-P.,Center dEtudes du Bouchet | Sharan M.,Indian Institute of Technology Delhi | Singh S.K.,Indian Institute of Technology Delhi
Pure and Applied Geophysics | Year: 2012

This paper addresses the parametric inverse problem of locating the point of release of atmospheric pollution. A finite set of observed mixing ratios is compared, by use of least squares, with the analogous mixing ratios computed by an adjoint dispersion model for all possible locations of the release. Classically, the least squares are weighted using the covariance matrix of the measurement errors. However, in practice, this matrix cannot be determined for the prevailing part of these errors arising from the limited representativity of the dispersion model. An alternative weighting proposed here is related to a unified approach of the parametric and assimilative inverse problems corresponding, respectively, to identification of the point of emission or estimation of the distributed emissions. The proposed weighting is shown to optimize the resolution and numerical stability of the inversion. The importance of the most common monitoring networks, with point detectors at various locations, is stressed as a misleading singular case. During the procedure it is also shown that a monitoring network, under given meteorological conditions, itself contains natural statistics about the emissions, irrespective of prior assumptions. © 2011 Springer Basel AG.


Sharan M.,Indian Institute of Technology Delhi | Issartel J.-P.,Center dEtudes du Bouchet | Singh S.K.,Indian Institute of Technology Delhi
Quarterly Journal of the Royal Meteorological Society | Year: 2012

An inversion technique is proposed to reconstruct an elevated point emission source of known height of atmospheric trace species from a finite number of concentration measurements in low-wind stable conditions. Observations from the SF 6 diffusion experiment at Idaho National Engineering Laboratory in low-wind stable conditions are used for the point source reconstruction. The source location and its strength are both retrieved exactly with the model-generated measurements in all the runs. With real data, the location is reconstructed with an average error of 20 m, mostly along the wind, and the intensity is retrieved within a factor of 2 in all the runs. The incurred errors in the retrieval are justified by estimating the angular departure between the vectors of measurement and adjoint function. A sensitivity study is carried out to analyse the sensitivity of the source estimation with respect to signal perturbation caused by the background concentration of the species in the ambient air. Source reconstruction is shown to be improved and more focused when the samplers having a measurement value corresponding to the background concentration are maintained in the inversion process. © 2012 Royal Meteorological Society.


Singh S.K.,Indian Institute of Technology Delhi | Sharan M.,Indian Institute of Technology Delhi | Issartel J.-P.,Center dEtudes du Bouchet
Boundary-Layer Meteorology | Year: 2013

An inverse modelling methodology is proposed for the identification of multiple-point sources releasing similar tracer, in which influences from the various emissions are merged in each detector's measurement. The identification is addressed from a limited merged set of atmospheric concentration measurements. The methodology is blended with the natural information provided by the geometry of the monitoring network in terms of the weight functions that interpret the associated visibility/illumination of the monitoring network. The release parameters, locations and intensities of the multiple-point sources are estimated by minimizing the objective function within the least squares framework. The methodology has been successfully applied to identify the two- and three- point simultaneous emissions from synthetic measurements generated by the model without noise or with controlled noise artificially added, and from pseudo-real measurements generated from the Indian Institute of Technology low wind diffusion experiment by combining several of single-point release runs. With the synthetic measurements, all the release parameters are retrieved exactly as those prescribed in all the runs. With the pseudo-real measurements, the release locations are retrieved with an average error of 13 m and intensities are estimated on an average within a factor of 1. 5. In a sensitivity analysis, it is shown that the incurred errors in the retrieval of the two- and three-point sources with the pseudo-real data correspond to the 10-30 % Gaussian distributed random noise in the observations. Theoretical and computational comparisons are given between the weighted and non-weighted classical formulations. In addition, an alternative strategy is proposed in order to reduce the computational time required in the source estimation. © 2012 Springer Science+Business Media B.V.


Kumar P.,University of Évry Val d'Essonne | Feiz A.-A.,University of Évry Val d'Essonne | Ngae P.,University of Évry Val d'Essonne | Singh S.K.,University of Évry Val d'Essonne | Issartel J.-P.,Center dEtudes du Bouchet
Atmospheric Environment | Year: 2015

An accurate simulation of the short-range plume dispersion of a hazardous pollutant in a geometrically complex urban region is a prerequisite in emergency preparedness and to assist regulators for developing effective policies. This study critically examines the real predictive capability of a three-dimensional Computational Fluid Dynamics (CFD) model, Fluidyn-PANACHE, to apply it in emergency contexts of an accidental or deliberate airborne release in urban regions. The model is evaluated with the Mock Urban Setting Test (MUST) field experiment of a continuous point source release in an idealized urban geometry of a regular array of shipping containers in various atmospheric stability varying from neutral to stable, and very stable conditions. The simulations are performed using three combinations (cases 1, 2, & 3) of inflow boundary conditions for wind and turbulence profiles. A detailed analysis with statistical measures shows that the performance of the Fluidyn-PANACHE against MUST experiment with all the three cases of the inflow boundary conditions is well achieved within the acceptable standards for air quality applications. The model with three cases 1, 2, & 3 predicts respectively 52.8%, 59.9%, and 67.9% of the total concentrations within a factor of two and shows an overall under-prediction. The sampling line maximum concentrations are better simulated by the CFD model with case-3 (95% within a factor of two) in comparison to other cases 1 & 2. A comparative statistical analysis is also performed with other evaluation studies in the literature for the averaged and sampling line maximum concentrations. The present evaluation of the Fluidyn-PANACHE strengthen the evidence that it is capable of dealing properly with the dispersion phenomena in geometrically complex urban environments. © 2015 Elsevier Ltd.


PubMed | Center dEtudes du Bouchet
Type: Journal Article | Journal: Toxicology in vitro : an international journal published in association with BIBRA | Year: 2010

Human keratinocyte cultures were treated with bis(betachloroethyl)sulphide (BCES), an alkylating and vesicant agent. At concentrations of 5 x 10(-4) to 5 x 10(-3)m, spontaneous detachment of the epithelium from the culture plate was observed, reproducing in vitro the cutaneous vesication observed in vivo. Progressive cellular alterations were shown with increasing concentrations of BCES (5 x 10(-5) to 5 x 10(-3)m). At low concentrations (5 x 10(-5)m), lesions of the nucleus, a significant target for BCES, were observed, along with lesions in the cytoplasmic organelles. An acute, dose-dependent depletion of cellular glutathione was observed, which occurred within 1 hr of treatment. Mechlorethamine, an analogue of BCES, induced at equivalent doses the same glutathione depletion and similar spontaneous detachment in vitro. We suggest that BCES, in addition to its genetic effects, acts by direct metabolic toxicity and induces glutathione depletion by direct conjugation. The lesions obtained in vitro reproduced those observed in vivo. Human keratinocyte cultures can be proposed as a good model for the study of the mechanisms of action of BCES.

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