RESUMO
The adaptation of Yersinia pestis, the flea-borne plague agent, to fluctuating environmental conditions is essential for the successful colonization of the flea vector. A previous comparative transcriptomic analysis showed that the Cpx pathway of Y. pestis is up-regulated in infected fleas. The CpxAR two-component system is a component of the envelope stress response and is critical for maintaining the integrity of the cell. Here, a phenotypic screening revealed a survival defect of the cpxAR mutant to oxidative stress and copper. The measured copper concentration in the digestive tract contents of fed fleas increased fourfold during the digestive process. By direct analysis of phosphorylation of CpxR by a Phos-Tag gel approach, we demonstrated that biologically relevant concentrations of copper triggered the system. Then, a competitive challenge highlighted the role of the CpxAR system in bacterial fitness during flea infection. Lastly, an in vitro sequential exposure to copper and then H2O2 to mimic the flea suggests a model in which, within the insect digestive tract, the CpxAR system would be triggered by copper, establishing an oxidative stress response. IMPORTANCE: The bacterium Yersinia pestis is the agent of flea-borne plague. Our knowledge of the mechanisms used by the plague bacillus to infect the flea vector is limited. The up-regulation of the envelope stress response under the control of the Cpx signaling pathway was previously shown in a transcriptomic study. Here, our in vivo and in vitro approaches suggest a model in which Y. pestis uses the CpxAR phosphorelay system to sense and respond to the copper present in the flea gut, thereby optimizing the flea gut colonization. In other words, the system is essential for bacterial fitness in the flea.
RESUMO
The Solidification/Stabilization method for dredged sediments remediation can be very effective for the immobilization of trace metal (TM) pollutants. The ordinary Portland cement (OPC) is largely used in this process. Nevertheless, the mechanical performance of treated sediments can be considerably improved at long term using ground granulated blast furnace slag (GGBS) as was shown in (Gutsalenko et al., 2018). Therefore, it is worth investigating whether this new binder is also relevant in terms of stabilization and mechanisms involved in this process. To meet this objective, leaching test, total attack, sequential extraction (SE) and X-ray Adsorption Near-Edge Structure (XANES) experiments measurements were performed. The results of the leaching tests conducted as part of the study are promising for the use of GGBS in the treatment of the real case project Dublin sediment and it outperforms the OPC-based treatment.. The sequential extraction method was applied to evaluate the potential risks of toxic elements according to their repartition in the Dublin sediment matrix and predicts the release of metals under different environmental conditions. It shows a lower perturbation rate of pollutants with the GGBS-based binder. Finally, XANES experiments demonstrate changes in the chemical environment of Zn and Cu after the treatment of the sediment with OPC compared to the GGBS rich binder. Consequently, this study finds that it is more pertinent to use the GGBS-based binder in terms of trace metal stabilization.
Assuntos
Poluentes Ambientais , Metais Pesados , Oligoelementos , Oligoelementos/análise , Metais , Metais Pesados/química , Sedimentos Geológicos/químicaRESUMO
Copper is both essential and toxic to living beings, which tightly controls its intracellular concentration. At the host-pathogen interface, copper is used by phagocytic cells to kill invading microorganisms. We investigated copper homeostasis in Bordetella pertussis, which lives in the human respiratory mucosa and has no environmental reservoir. B. pertussis has considerably streamlined copper homeostasis mechanisms relative to other Gram-negative bacteria. Its single remaining defense line consists of a metallochaperone diverted for copper passivation, CopZ, and two peroxide detoxification enzymes, PrxGrx and GorB, which together fight stresses encountered in phagocytic cells. Those proteins are encoded by an original, composite operon assembled in an environmental ancestor, which is under sensitive control by copper. This system appears to contribute to persistent infection in the nasal cavity of B. pertussis-infected mice. Combining responses to co-occurring stresses in a tailored operon reveals a strategy adopted by a host-restricted pathogen to optimize survival at minimal energy expenditure.
Assuntos
Bordetella pertussis/metabolismo , Cobre/metabolismo , Óperon , Bordetella bronchiseptica/metabolismo , Bordetella pertussis/genética , Homeostase , Peróxidos/metabolismoRESUMO
The performances of five Diffusive Gradients in Thin Films (DGT) binding gels, namely 3-mercapto-functionalized silica (3MP), ferrihydrite (Fh), Metsorb®, zinc ferrite (ZnFe2O4), and Zirconium oxide (ZrO2), were evaluated for in situ determination of As speciation in water and sediments. A combination of batch experiments at various pH (without addition of buffers) and in the presence of reduced species (Mn2+, Fe2+ and HS-),time-series experiments in oxic waters, and in situ deployment in anoxic river sediments has permitted to evaluate the potential interferences among the binding gels. Firstly, the efficiency of each DGT binding gel dedicated to total As (i.e., Fh, Metsorb®, ZnFe2O4 and ZrO2) or As(III) (i.e., 3MP) determination confirms that the determination of As species is possible in oxic freshwater and seawater over 96â¯h for a wide range of pH (5-9). Secondly, concerning the deployment in river sediment, high HCO3- concentrations have a little negative effect only on the DGT performances of the iron(III)-binding gels (i.e, Fh and ZnFe2O4). Thirdly, the presence of sulfides does not show any effect on the DGT uptake of As, but strongly affects the elution factor parameter. Discrepancies in elution between the different binding gels potentially result in precipitation of orpiment, especially in 1â¯molâ¯L-1 HNO3. A correction of the classical elution factor derived from batch experiments was applied to provide more representative results. Finally, this study shows the difficulties to determine As speciation in anoxic sediments, and suggests that corrections of the elution factor may be required as a function of the species present in the deployment matrices.