Nicolas Menguy

Size-, Composition- and Shape-Dependent Toxicological Impact of Metal Oxide Nanoparticles and Carbon Nanotubes toward Bacteria

Ecotoxicological effects of nanoparticles (NP) are still poorly documented while their commercialization for industrial and household applications increases. The aim of this study was to evaluate the influence of physicochemical characteristics on metal oxide NP and carbon nanotubes toxicological effects toward bacteria. Two strains of bacteria, Cupriavidus metallidurans CH34 and Escherichia coli MG1655 were exposed to TiO(2) or Al(2)O(3) NP or to multiwalled-carbon nanotubes (MWCNT). Particular attention was paid on optimizing NP dispersion to obtain nonagglomerated suspensions. Our results show that NP toxicity depends on their chemical composition, size, surface charge, and shape but not on their crystalline phase. MWCNT toxicity does not depend on their purity. Toxicity also depends on the bacterial strain: E. coli MG1655 is sensitive to NP, whereas C. metallidurans CH34 is not. Interestingly, NP are accumulated in both bacterial strains, and association between NP and bacteria is necessary for bacterial death to occur. NP may then represent a danger for the environment, causing the disappearance of some sensitive bacterial strains such as E. coli MG1655, but also being mobilized by nonsensitive strains such as C. metallidurans CH34 and transported through the whole ecosystem.

Magnetotactic bacteria form magnetite from a phosphate-rich ferric hydroxide via nanometric ferric (oxyhydr)oxide intermediates

The iron oxide mineral magnetite (Fe3O4) is produced by various organisms to exploit magnetic and mechanical properties. Magnetotactic bacteria have become one of the best model organisms for studying magnetite biomineralization, as their genomes are sequenced and tools are available for their genetic manipulation. However, the chemical route by which magnetite is formed intracellularly within the so-called magnetosomes has remained a matter of debate. Here we used X-ray absorption spectroscopy at cryogenic temperatures and transmission electron microscopic imaging techniques to chemically characterize and spatially resolve the mechanism of biomineralization in those microorganisms. We show that magnetite forms through phase transformation from a highly disordered phosphate-rich ferric hydroxide phase, consistent with prokaryotic ferritins, via transient nanometric ferric (oxyhydr)oxide intermediates within the magnetosome organelle. This pathway remarkably resembles recent results on synthetic magnetite formation and bears a high similarity to suggested mineralization mechanisms in higher organisms.

Environmental parameters affect the physical properties of fast-growing magnetosomes

Abstract Magnetotactic bacteria are known to mediate the formation of intracellular magnetic nanoparticles in organelles called magnetosomes. These magnetite crystals are formed through a process called biologically controlled mineralization, in which the microorganisms exert a strict control over the formation and development of the mineral phase. By inducing magnetite nucleation and growth in resting, Fe-starved cells of Magnetospirillum gryphiswaldense, we have followed the dynamics of magnetosome development. By studying the properties of the crystals at several steps of maturity, we observed that freshly induced particles lacked a well-defined morphology. More surprisingly, although the mean particle size of mature magnetosomes is similar to that of magnetosomes formed by constantly growing and Fe-supplemented bacteria, we found that other physical properties such as crystal-size distribution, aspect ratio, and morphology significantly differ. Correlating these results with measurements of Fe uptake rates, we suggest that the expression of different faces is favored for different growth conditions. These results imply that the biological control over magnetite biomineralization by magnetotactic bacteria can be disturbed by environmental parameters. Specifically, the morphology of magnetite crystals is not exclusively determined by biological intervention through vectorial regulation at the organic boundaries or by molecular interaction with the magnetosome membrane, but also by the rates of Fe uptake. This insight may contribute to better define biomarkers and to an improved understanding of biomineralizing systems.

Direct and indirect CeO2 nanoparticles toxicity for Escherichia coli and Synechocystis

Abstract Physico-chemical interactions between nanoparticles and cell membranes play a crucial role in determining the cytotoxicity of nanoparticles, which may thereby vary depending on the nature of the target microorganisms. We investigated the responses of two different models of unicellular bacteria to cerium oxide (CeO2) nanoparticles. These organisms are: Synechocystis PCC6803 a representative of environmentally important cyanobacterial organisms (producer of biomass for aquatic food chains), and Escherichia coli a representative of intestine-colonizing bacteria. Coupling physico-chemical (adsorption isotherms and electrophoretic mobility), biological (survival tests), microscopical (SEM, TEM and EDS) and spectroscopic (XANES) methods, we enlightened two distinct mechanisms for the CeO2 nanoparticles toxicological impact: A ‘direct’ mechanism that requires a close contact between nanoparticles and cell membranes, and an ‘indirect’ influence elicited by the acidity of nanoparticles stabilizing agents. We showed that E. coli is sensitive to the ‘direct’ effects of nanoparticles, whereas Synechocystis being protected by extracellular polymeric substances preventing direct cellular contacts is sensitive only to the ‘indirect’ mechanism. Consequently, our findings demonstrate the importance of the ‘direct/indirect’ effects of nanoparticles on cell fitness, a phenomenon that should be systematically investigated with appropriate techniques and dose metrics to make meaningful environmental and/or health recommendations.

Nanobacteria-like calcite single crystals at the surface of the Tataouine meteorite

Nanobacteria-like objects evidenced at the surface of the orthopyroxenes of the Tataouine meteorite in South Tunisia have been studied by scanning and transmission electron microscopies. A method of micromanipulation has been developed to ensure that exactly the same objects were studied by both methods. We have shown that the nanobacteria-like objects are spatially correlated with filaments of microorganisms that colonized the surface of the meteoritic pyroxene during its 70 years of residence in the aridic Tataouine soil. Depressions of a few micrometers in depth are observed in the pyroxene below the carbonates, indicating preferential dissolution of the pyroxene and calcite precipitation at these locations. The nanobacteria-like small rods that constitute calcium carbonate rosettes are well crystallized calcite single crystals surrounded by a thin amorphous layer of carbonate composition that smoothes the crystal edges and induces rounded shapes. Those morphologies are unusual for calcite single crystals observed in natural samples. A survey of recent literature suggests that the intervention of organic compounds derived from biological activity is likely in their formation.

EXAFS and HRTEM evidence for As(III)-containing surface precipitates on nanocrystalline magnetite: implications for As sequestration.

Arsenic sorption onto iron oxide spinels such as magnetite could contribute to immobilization of arsenite (AsO3(3-)), the reduced, highly toxic form of arsenic in contaminated anoxic groundwaters, as well as to putative remediation processes. Nanocrystalline magnetite (<20 nm) is known to exhibit higher efficiency for arsenite sorption than larger particles, sorbing as much as approximately 20 micromol/m2 of arsenite. To improve our understanding of this process, we investigated the molecular level structure of As(III)-containing sorption products on two types of fine-grained magnetite: (1) a biogenic one with an average particle diameter of 34 nm produced by reduction of lepidocrocite (gamma-FeOOH) by Shewanella putrefaciens and (2) a synthetic, abiotic, nanocrystalline magnetite with an average particle diameter of 11 nm. Results from extended X-ray absorption spectroscopy (EXAFS) for both types of magnetite with As(III) surface coverages of up to 5 micromol/m2 indicate that As(III) forms dominantly inner-sphere, tridentate, hexanuclear, corner-sharing surface complexes (3C) in which AsO3 pyramids occupy vacant tetrahedral sites on octahedrally terminated {111} surfaces of magnetite. Formation of this type of surface complex results in a decrease in dissolved As(III) concentration below the maximum concentration level recommended by the World Health Organization (10 microg/L), which corresponds to As(III) surface coverages of 0.16 and 0.19 micromol/m2 in our experiments. In addition, high-resolution transmission electron microscopy (HRTEM) coupled with energy dispersive X-ray spectroscopy (EDXS) analyses revealed the occurrence of an amorphous As(III)-rich surface precipitate forming at As(III) surface coverages as low as 1.61 micromol/m2. This phase hosts the majority of adsorbed arsenite at surface coverages exceeding the theoretical maximum site density of vacant tetrahedral sites on the magnetite {111} surface (3.2 sites/nm2 or 5.3 micromol/m2). This finding helps to explain the exceptional As(III) sorption capacity of nanocrystalline magnetite particles (>10 micromol/m2). However, the higher solubility of the amorphous surface precipitate compared to the 3C surface complexes causes a dramatic increase of dissolved As concentration for coverages above 1.9 micromol/m2.

Intracellular Ca-carbonate biomineralization is widespread in cyanobacteria.

Significance Cyanobacteria are known to promote the precipitation of Ca-carbonate minerals by the photosynthetic uptake of inorganic carbon. This process has resulted in the formation of carbonate deposits and a fossil record of importance for deciphering the evolution of cyanobacteria and their impact on the global carbon cycle. Though the mechanisms of cyanobacterial calcification remain poorly understood, this process is invariably thought of as extracellular and the indirect by-product of metabolic activity. Here, we show that contrary to common belief, several cyanobacterial species perform Ca-carbonate biomineralization intracellularly. We observed at least two phenotypes for intracellular biomineralization, one of which shows an original connection with cell division. These findings open new perspectives on the evolution of cyanobacterial calcification. Cyanobacteria have played a significant role in the formation of past and modern carbonate deposits at the surface of the Earth using a biomineralization process that has been almost systematically considered induced and extracellular. Recently, a deep-branching cyanobacterial species, Candidatus Gloeomargarita lithophora, was reported to form intracellular amorphous Ca-rich carbonates. However, the significance and diversity of the cyanobacteria in which intracellular biomineralization occurs remain unknown. Here, we searched for intracellular Ca-carbonate inclusions in 68 cyanobacterial strains distributed throughout the phylogenetic tree of cyanobacteria. We discovered that diverse unicellular cyanobacterial taxa form intracellular amorphous Ca-carbonates with at least two different distribution patterns, suggesting the existence of at least two distinct mechanisms of biomineralization: (i) one with Ca-carbonate inclusions scattered within the cell cytoplasm such as in Ca. G. lithophora, and (ii) another one observed in strains belonging to the Thermosynechococcus elongatus BP-1 lineage, in which Ca-carbonate inclusions lie at the cell poles. This pattern seems to be linked with the nucleation of the inclusions at the septum of the cells, showing an intricate and original connection between cell division and biomineralization. These findings indicate that intracellular Ca-carbonate biomineralization by cyanobacteria has been overlooked by past studies and open new perspectives on the mechanisms and the evolutionary history of intra- and extracellular Ca-carbonate biomineralization by cyanobacteria.

The iron status in colloidal matter from the Rio Negro, Brasil

Colloidal and particulate natural organic matter was size-fractionated and concentrated from the Rio Negro river using tangential-flow filtration (TFF). Flow field-flow fractionation (F-FFF), with UV absorbance detection (UVA), was used to investigate the molecular weight distributions of the organic colloids. To further characterize the nature of the Rio Negro colloids, the size distributions of the iron concentrations were determined by direct on-line coupling of the F-FFF to an inductively coupled plasma emission spectrometer (ICP-AES). The size distributions obtained by both F-FFF-UVA and F-FFF-ICP-AES were considerably smaller than expected from the stated pore size of the TFF membranes. These results demonstrate that care must be taken in using TFF to classify the size distribution of organic colloids and associated elements present in rivers. The iron distribution is more closely correlated to the organic matter distribution in the colloidal fraction than in the particulate fraction, but it is shifted towards heavier molecular weights for both fractions. The combination of electron paramagnetic resonance spectroscopy (EPR) and transmission electron microscopy (TEM) reveals the presence of iron occurring as: specific complexes with organic functional groups, as Fe-oxide/oxihydroxide phases, or as structurally incorporated component in kaolinite. Particulate and colloidal fractions are differentiated from each other with respect to the iron forms, which is in qualitative agreement with the F-FFF-ICP-AES results.

Experimental investigation of the stability of Fe-rich carbonates in the lower mantle

The fate of carbonates in the Earths mantle plays a key role in the geodynamical carbon cycle. Although iron is a major component of the Earths lower mantle, the stability of Fe-bearing carbonates has rarely been studied. Here we present experimental results on the stability of Fe-rich carbonates at pressures ranging from 40 to 105 GPa and temperatures of 1450-3600 K, corresponding to depths within the Earths lower mantle of about 1000-2400 km. Samples of iron oxides and iron-magnesium oxides were loaded into CO2 gas and laser heated in a diamond-anvil cell. The nature of crystalline run products was determined in situ by X-ray diffraction, and the recovered samples were studied by analytical transmission electron microscopy and scanning transmission X-ray microscopy. We show that Fe-(II) is systematically involved in redox reactions with CO2 yielding to Fe-(III)-bearing phases and diamonds. We also report a new Fe-(III)-bearing high-pressure phase resulting from the transformation of FeCO3 at pressures exceeding 40 GPa. The presence of both diamonds and an oxidized C-bearing phase suggests that oxidized and reduced forms of carbon might coexist in the deep mantle. Finally, the observed reactions potentially provide a new mechanism for diamond formation at great depth.

Evidence of Double-Walled Al―Ge Imogolite-Like Nanotubes. A Cryo-TEM and SAXS Investigation

It has been recently discovered that the synthesis of Al-Ge imogolite-like nanotubes is possible at high concentration. Despite this initial success, the structure of these Al-Ge imogolite-like nanotubes remains not completely understood. Using high resolution cryo-TEM and Small Angle X-ray Scattering, we unravel their mesoscale structure in two contrasted situations. On the one hand, Al-Ge imogolite nanotubes synthesized at 0.25 M are double-walled nanotubes of 4.0 +/- 0.1 nm with an inner tube of 2.4 +/- 0.1 nm. Moreover, SAXS data also suggest that the two concentric tubes have an equal length and identical wall structure. On the other hand, at higher concentration (0.5M), both SAXS and cryo-TEM data confirm the formation of single-walled nanotubes of 3.5 +/- 0.15 nm. Infrared spectroscopy confirms the imogolite structure of the tubes. This is the first evidence of any double-walled imogolite or imogolite-like nanotubes likely to renew interest in these materials and associated potential applications.