Astrid Avellan

Nanoparticle Uptake in Plants: Gold Nanomaterial Localized in Roots of Arabidopsis thaliana by X-ray Computed Nanotomography and Hyperspectral Imaging

Terrestrial plants can internalize and translocate nanoparticles (NPs). However, direct evidence for the processes driving the NP uptake and distribution in plants is scarce at the cellular level. Here, NP-root interactions were investigated after 10 days of exposure of Arabidopsis thaliana to 10 mg·L-1 of negatively or positively charged gold NPs (∼12 nm) in gels. Two complementary imaging tools were used: X-ray computed nanotomography (nano-CT) and enhanced dark-field microscopy combined with hyperspectral imaging (DF-HSI). The use of these emerging techniques improved our ability to detect and visualize NP in plant tissue: by spectral confirmation via DF-HSI, and in three dimensions via nano-CT. The resulting imaging provides direct evidence that detaching border-like cells (i.e., sheets of border cells detaching from the root) and associated mucilage can accumulate and trap NPs irrespective of particle charge. On the contrary, border cells on the root cap behaved in a charge-specific fashion: positively charged NPs induced a higher mucilage production and adsorbed to it, which prevented translocation into the root tissue. Negatively charged NPs did not adsorb to the mucilage and were able to translocate into the apoplast. These observations provide direct mechanistic insight into NP-plant interactions, and reveal the important function of border cells and mucilage in interactions of plants with charged NPs.

Remote Biodegradation of Ge–Imogolite Nanotubes Controlled by the Iron Homeostasis of Pseudomonas brassicacearum

The toxicity of high-aspect-ratio nanomaterials (HARNs) is often associated with oxidative stress. The essential nutrient Fe may also be responsible of oxidative stress through the production of reactive oxygen species. In the present study, it has been examined to what extent adding Fenton reaction promoting Fe impacted the toxicity of an alumino-germanate model HARN. Structural addition of only 0.95% wt Fe to Ge-imogolite not only alleviated the toxicity observed in the case of Fe-free nanotubes but also stimulated bacterial growth. This was attributed to the metabolization of siderophore-mobilized Fe from the nanotube structure. This was evidenced by the regulation of the homeostasis-monitoring intracellular Fe levels. This was accompanied by a biodegradation of the nanotubes approaching 40%, whereas the Fe-free nanomaterial remained nearly untouched.

Disruption of Autolysis in Bacillus subtilis using TiO 2 Nanoparticles

In contrast to many nanotoxicity studies where nanoparticles (NPs) are observed to be toxic or reduce viable cells in a population of bacteria, we observed that increasing concentration of TiO2 NPs increased the cell survival of Bacillus subtilis in autolysis-inducing buffer by 0.5 to 5 orders of magnitude over an 8 hour exposure. Molecular investigations revealed that TiO2 NPs prevent or delay cell autolysis, an important survival and growth-regulating process in bacterial populations. Overall, the results suggest two potential mechanisms for the disruption of autolysis by TiO2 NPs in a concentration dependent manner: (i) directly, through TiO2 NP deposition on the cell wall, delaying the collapse of the protonmotive-force and preventing the onset of autolysis; and (ii) indirectly, through adsorption of autolysins on TiO2 NP, limiting the activity of released autolysins and preventing further lytic activity. Enhanced darkfield microscopy coupled to hyperspectral analysis was used to map TiO2 deposition on B. subtilis cell walls and released enzymes, supporting both mechanisms of autolysis interference. The disruption of autolysis in B. subtilis cultures by TiO2 NPs suggests the mechanisms and kinetics of cell death may be influenced by nano-scale metal oxide materials, which are abundant in natural systems.

Influence of structural defects of Ge-imogolite nanotubes on their toxicity towards Pseudomonas brassicacearum

While the definition of a nanomaterial (NM) is mainly based on size, it is known that decreasing size can induce structural modifications to compensate for increased surface energy. Nevertheless, the influence of these structural modifications on NM toxicity, and in particular structural defects, is poorly studied mainly because of the difficulty in varying the crystallinity of a NM without changing any other morphological parameters. In this study, we used a single-walled alumino-germanate nanotube (Ge-imogolite) as a model, for which this can be achieved. Differences in toxicity of well-crystallized Ge-imogolite vs. Ge-imogolite presenting vacant sites towards the soil bacteria Pseudomonas brassicacearum were studied. Well crystallized tubes led to moderate toxicity attributed to a direct contact with the bacteria and the generation of reactive oxygen species, whereas tubes presenting vacant sites caused more severe toxic effects without any direct contact nor ROS generation. The bacterial growth inhibition in the presence of lacunar tubes was attributed to indirect mechanisms as their higher solubility leads to Al or Ge toxic ion release and/or to the retention of essential nutrients on the vacancies. This study highlights the close correlation between structural defects of a nanomaterial and the modulation of its mechanisms of toxicity.

Speciation of Mercury in Selected Areas of the Petroleum Value Chain

Petroleum, natural gas, and natural gas condensate can contain low levels of mercury (Hg). The speciation of Hg can affect its behavior during processing, transport, and storage so efficient and safe management of Hg requires an understanding of its chemical form in oil, gas and byproducts. Here, X-ray absorption spectroscopy was used to determine the Hg speciation in samples of solid residues collected throughout the petroleum value chain including stabilized crude oil residues, sediments from separation tanks and condensate glycol dehydrators, distillation column pipe scale, and biosludge from wastewater treatment. In all samples except glycol dehydrators, metacinnabar (β-HgS) was the primary form of Hg. Electron microscopy on particles from a crude sediment showed nanosized (<100 nm) particles forming larger aggregates, and confirmed the colocalization of Hg and sulfur. In sediments from glycol dehydrators, organic Hg(SR)2 accounted for ∼60% of the Hg, with ∼20% present as β-HgS and/or Hg(SR)4 species. β-HgS was the predominant Hg species in refinery biosludge and pipe scale samples. However, the balance of Hg species present in these samples depended on the nature of the crude oil being processed, i.e. sweet (low sulfur crudes) vs sour (higher sulfur crudes). This information on Hg speciation in the petroleum value chain will inform development of better engineering controls and management practices for Hg.

Non-linear release dynamics for a CeO 2 nanomaterial embedded in a protective wood stain, due to matrix photo-degradation

The release of CeO2-bearing residues during the weathering of an acrylic stain enriched with CeO2 nanomaterial designed for wood protection (Nanobyk brand additive) was studied under two different scenarios: (i) a standard 12-weeks weathering protocol in climate chamber, that combined condensation, water spraying and UV-visible irradiation and (ii) an alternative accelerated 2-weeks leaching batch assay relying on the same weathering factors (water and UV), but with a higher intensity of radiation and immersion phases. Similar Ce released amounts were evidenced for both scenarios following two phases: one related to the removal of loosely bound material with a relatively limited release, and the other resulting from the degradation of the stain, where major release occurred. A non-linear evolution of the release with the UV dose was evidenced for the second phase. No stabilization of Ce emissions was reached at the end of the experiments. The two weathering tests led to different estimates of long-term Ce releases, and different degradations of the stain. Finally, the photo-degradations of the nanocomposite, the pure acrylic stains and the Nanobyk additive were compared. The incorporation of Nanobyk into the acrylic matrix significantly modified the response of the acrylic stain to weathering.

Effect of Initial Speciation of Copper- and Silver-Based Nanoparticles on Their Long-Term Fate and Phytoavailability in Freshwater Wetland Mesocosms

Ag0- and CuO-engineered nanomaterials (ENMs) or their sulfidized forms are introduced into freshwater wetlands through wastewater effluent and agricultural runoff. Knowledge about the rates of transformations of these ENMs in realistic environments and the impact of the form of the incoming ENM (i.e., sulfidized or pristine) on bioavailability and fate is limited. Here, five freshwater wetland mesocosms were exposed to 3 g of total metal as CuO, CuS, Ag0, or Ag2S ENMs or soluble CuNO3 added weekly for 1 month. Total metal and metal speciation was measured in sediment and plant samples collected 1, 3, 6, and 9 months after addition. The form of the added ENM did not affect the metal distribution, and ENMs distributed similarly to added ionic Cu or Ag. For the dosing condition used, ∼50% of the added Ag or Cu metal mass was found in Egeria densa plant tissue, with the remainder primarily in the surficial sediment. Ag0 and CuO ENMs transformed quickly in sediment, with no evidence of CuO and only ∼4% of silver present as Ag0 ENM 1 week after the last ENM addition. In contrast to sediment, Ag0 and CuO ENMs were persistent in E. densa tissues for up to 9 and 6 months, respectively. The persistence of ENMs in E. densa suggests that chronic exposures, or food web transfers, for both the transformed and the initially added ENMs are possible.

Gold nanoparticle biodissolution by a freshwater macrophyte and its associated microbiome

Predicting nanoparticle fate in aquatic environments requires mimicking of ecosystem complexity to observe the geochemical processes affecting their behaviour. Here, 12 nm Au nanoparticles were added weekly to large-scale freshwater wetland mesocosms. After six months, ~70% of Au was associated with the macrophyte Egeria densa, where, despite the thermodynamic stability of Au0 in water, the pristine Au0 nanoparticles were fully oxidized and complexed to cyanide, hydroxyls or thiol ligands. Extracted biofilms growing on E. densa leaves were shown to dissolve Au nanoparticles within days. The Au biodissolution rate was highest for the biofilm with the lowest prevalence of metal-resistant taxa but the highest ability to release cyanide, known to promote Au0 oxidation and complexation. Macrophytes and the associated microbiome thus form a biologically active system that can be a major sink for nanoparticle accumulation and transformations. Nanoparticle biotransformation in these compartments should not be ignored, even for nanoparticles commonly considered to be stable in the environment.Gold nanoparticles typically considered inert in oxic waters accumulate in freshwater wetland subaquatic plants and are completely biotransformed to oxidized Au species by the associated cyanogenic biofilm.

Temporal evolution of copper distribution and speciation in roots of Triticum aestivum exposed to CuO, Cu(OH)2, and CuS nanoparticles

Utilization of nanoparticles (NP) in agriculture as fertilizers or pesticides requires an understanding of the NP properties influencing their interactions with plant roots. To evaluate the influence of the solubility of Cu-based NP on Cu uptake and NP association with plant roots, wheat seedlings were hydroponically exposed to 1 mg/L of Cu NPs with different solubilities [CuO, CuS, and Cu(OH)2] for 1 h then transferred to a Cu-free medium for 48 h. Fresh, hydrated roots were analyzed using micro X-ray fluorescence (μ-XRF) and imaging fluorescence X-ray absorption near edge spectroscopy (XANES imaging) to provide laterally resolved distribution and speciation of Cu in roots. Higher solubility Cu(OH)2 NPs provided more uptake of Cu after 1 h of exposure, but the lower solubility materials (CuO and CuS) were more persistent on the roots and continued to deliver Cu to plant leaves over the 48 h depuration period. These results demonstrate that NPs, by associating to the roots, have the potential to play a role in slowly providing micronutrients to plants. Thus, tuning the solubility of NPs may provide a long-term slow delivery of micronutrients to plants and provide important information for understanding mechanisms responsible for plant uptake, transformation, and translocation of NPs.

Size-Based Differential Transport, Uptake, and Mass Distribution of Ceria (CeO2) Nanoparticles in Wetland Mesocosms

Trace metals associated with nanoparticles are known to possess reactivities that are different from their larger-size counterparts. However, the relative importance of small relative to large particles for the overall distribution and biouptake of these metals is not as well studied in complex environmental systems. Here, we have examined differences in the long term fate and transport of ceria (CeO2) nanoparticles of two different sizes (3.8 vs 185 nm), dosed weekly to freshwater wetland mesocosms over 9 months. While the majority of CeO2 particles were detected in soils and sediments at the end of nine months, there were significant differences observed in fate, distribution, and transport mechanisms between the two materials. Small nanoparticles were removed from the water column primarily through heteroaggregation with suspended solids and plants, while large nanoparticles were removed primarily by sedimentation. A greater fraction of small particles remained in the upper floc layers of sediment relative to the large particles (31% vs 7%). Cerium from the small particles were also significantly more bioavailable to aquatic plants (2% vs 0.5%), snails (44 vs 2.6 ng), and insects (8 vs 0.07 μg). Small CeO2 particles were also significantly reduced from Ce(IV) to Ce(III), while aquatic sediments were a sink for untransformed large nanoparticles. These results demonstrate that trace metals originating from nanoscale materials have much greater potential than their larger counterparts to distribute throughout multiple compartments of a complex aquatic ecosystem and contribute to the overall bioavailable pool of the metal for biouptake and trophic transfer.