Metabolomic and proteomic investigations of impacts of titanium dioxide nanoparticles on Escherichia coli.

In a previous study, it was demonstrated that the toxic impact of titanium dioxide nanoparticles on Escherichia coli starts at 10 ppm and is closely related to the presence of little aggregates. It was also assumed that only a part of the bacterial population is able to adapt to this stress and attempts to survive. Proteomic analyses, supported by results from metabolomics, reveal that exposure of E. coli to nano-TiO2 induces two main effects on bacterial metabolism: firstly, the up-regulation of proteins and the increase of metabolites related to energy and growth metabolism; secondly, the down-regulation of other proteins resulting in an increase of metabolites, particularly amino acids. Some proteins, e.g. chaperonin 1 or isocitrate dehydrogenase, and some metabolites, e.g. phenylalanine or valine, might be used as biomarkers of nanoparticles stress. Astonishingly, the ATP content gradually rises in relation with the nano-TiO2 concentration in the medium, indicating a dramatic release of ATP by the damaged cells. These apparently contradictory results accredit the thesis of a heterogeneity of the bacterial population. This heterogeneity is also confirmed by SEM images which show that while some bacteria are fully covered by nano-TiO2, the major part of the bacterial population remains free from nanoparticles, resulting in a difference of proteome and metabolome. The use of combined-omics has allowed to better understand the heterogeneous bacterial response to nano-TiO2 stress due to heterogeneous contacts between the protagonists under environmental conditions.

Prospective analysis of the flows of certain rare earths in Europe at the 2020 horizon.

This paper proposes a forecast of certain rare earth flows in Europe at the 2020 horizon, based on an analysis of trends influencing various actors of the rare earth industry along the value chain. While 2020 is indicated as the forecast horizon, the analysis should be considered as more representative of the next decade. The rare earths considered here are used in applications that are important for a low-carbon energy transition and/or have a significant recycling potential: NdFeB magnets (Pr, Nd, Dy), NiMH batteries (Pr, Nd) and fluorescent lamp phosphors (Eu, Tb, Y). An analysis of major trends affecting the rare earth industry in Europe along the value chain (including extraction, separation, fabrication, manufacture, use and recycling), helps to build a scenario for a material flow analysis of these rare earths in Europe. The scenario assumes in particular that during the next decade, there exists a rare earth mine in production in Europe (with Norra Kärr in Sweden as a most likely candidate) and also that recycling is in line with targets proposed in recent European legislation. Results are presented in the form of Sankey diagrams which help visualize the various flows for the three applications. For example, calculations forecast flows from extraction to separation of Pr, Nd and Dy for magnet applications in Europe, on the order of 310 tons, 980 tons and 80 tons rare earth metal resp., while recycled flows are 35 tons, 110 tons and 30 tons resp. Calculations illustrate how the relative contribution of recycling to supply strongly depends on the situation with respect to demand. Considering the balance between supply and demand, it is not anticipated any significant shortage of rare earth supply in Europe at the 2020 horizon, barring any new geopolitical crisis involving China. For some heavy rare earths, supply will in fact largely outweigh demand, as for example Europium due to the phasing out of fluorescent lights by LEDs.

Exopolysaccharides protect Synechocystis against the deleterious effects of titanium dioxide nanoparticles in natural and artificial waters.

We have studied the effect of TiO2 nanoparticles (NPs) on the model cyanobacteria Synechocystis PCC6803. We used well-characterized NPs suspensions in artificial and natural (Seine River, France) waters. We report that NPs trigger direct (cell killing) and indirect (cell sedimentation precluding the capture of light, which is crucial to photosynthesis) deleterious effects. Both toxic effects increase with NPs concentration and are exacerbated by the presence of UVAs that increase the production of Reactive Oxygen Species (hydroxyl and superoxide radicals) by TiO2 NPs. Furthermore, we compared the responses of the wild-type strain of Synechocystis, which possesses abundant exopolysaccharides surrounding the cells, to that of an EPS-depleted mutant. We show, for the first time, that the exopolysaccharides play a crucial role in Synechocystis protection against cell killing caused by TiO2 NPs.

Influence of exopolysaccharides on the electrophoretic properties of the model cyanobacterium Synechocystis.

The influence of extracellular polymeric substances (EPS) on cell electrokinetics was investigated in the model cyanobacterium Synechocystis, in wild-type strains and in ten EPS-depleted mutants. The charge density and the softness of the EPS polyelectrolyte layer were calculated from the dependence of the electrophoretic mobility values of the cells with the ionic strength of the surrounding fluid. Electrophoretic mobility data showed that the eleven Synechocystis strains investigated behave as soft particles and cannot be adequately described by classical electrokinetic models of rigid particles. EPS surrounding the cells, especially those released in the growth medium, significantly increased the softness of the cell surface. Furthermore, the anionic nature of EPS resulted in negative surface charge densities, which appeared to be strongly dependent on the composition of the suspending fluid, as documented by a strong reduction of their absolute values in the presence of calcium cations. These finding stresses the importance of the physicochemical properties of EPS and cell surfaces of cyanobacteria, for both cell-to-medium and cell-to-cell communications. In turn, these results emphasize that, whenever possible, natural waters should be used for meaningful ecotoxicological analyses of potential toxics.

Multidisciplinary evidences that Synechocystis PCC6803 exopolysaccharides operate in cell sedimentation and protection against salt and metal stresses.

Little is known about the production of exopolysaccharides (EPS) in cyanobacteria, and there are no genetic and physiological evidences that EPS are involved in cell protection against the frequently encountered environmental stresses caused by salt and metals. We studied four presumptive EPS production genes, sll0923, sll1581, slr1875 and sll5052, in the model cyanobacterium Synechocystis PCC6803, which produces copious amounts of EPS attached to cells (CPS) and released in the culture medium (RPS) as shown here. We show that sll0923, sll1581, slr1875 and sll5052 are all dispensable to the growth of all corresponding single and double deletion mutants in absence of stress. Furthermore, we report that sll0923, sll1581 and slr1875 unambiguously operate in the production of both CPS and RPS. Both sll1581 and slr1875 are more important than sll0923 for CPS production, whereas the contrary is true for RPS production. We show that the most EPS-depleted mutant, doubly deleted for sll1581 and slr1875, lacks the EPS mantle that surrounds WT cells and sorbs iron in their vicinity. Using this mutant, we demonstrate for the first time that cyanobacterial EPS directly operate in cell protection against NaCl, CoCl(2), CdSO(4) and Fe-starvation. We believe that our EPS-depleted mutants will be useful tools to investigate the role of EPS in cell-to-cell aggregation, biofilm formation, biomineralization and tolerance to environmental stresses. We also suggest using the fast sedimenting mutants as biotechnological cell factories to facilitate the otherwise expensive harvest of the producer cell biomass and/or its separation from products excreted in the growth media.

Interaction between Escherichia coli and TiO2 nanoparticles in natural and artificial waters.

Seine River water was used as a natural environmental medium to quantify the ecotoxicological impact of three types of manufactured titanium dioxide (TiO(2)) nanoparticles toward the model bacterium Escherichia coli. Under ambient light, a significant toxicity starting at 10 ppm of TiO(2) in water was observed. Presence of the anatase polymorph slightly increased the toxicity in comparison to pure rutile samples. Furthermore, the toxicity was found to be lower at pH 5 compared to Seine water (pH 8). To assess the nanoparticles state of dispersion and their interactions with bacteria, cryogenic transmission electron microscopy (TEM) and zeta potential measurements were performed. A higher sorption of nanoparticle aggregates on cells is observed at pH 5 compared to Seine water. This allows concluding that the observed toxicity is not directly linked to the particles sorption onto the cell surfaces. In spite of stronger interaction between cells and nanoparticles at pH 5, a bacterial subpopulation apparently non-interacting with nanoparticles is evidenced by both TEM and zeta potential measurements. Such heterogeneities in cell populations can increase global bacterial resistance to TiO(2) nanoparticles.