Bioaccumulation of CeO2 Nanoparticles by Earthworms in Biochar-Amended Soil: A Synchrotron Microspectroscopy Study.

The interactions of nanoparticles (NPs) with biochar and soil components may substantially influence NP availability and toxicity to biota. In the present study, earthworms ( Eisenia fetida) were exposed for 28 days to a residential or agricultural soil amended with 0-2000 mg of CeO2 NP/kg and with biochar (produced by the pyrolysis of pecan shells at 350 and 600 °C) at various application rates [0-5% (w/w)]. After 28 days, earthworms were depurated and analyzed for Ce content, moisture content, and lipid peroxidation. The results showed minimal toxicity to the worms; however, biochar (350 or 600 °C) was the dominant factor, accounting for 94 and 84% of the variance for the moisture content and lipid peroxidation, respectively, in the exposed earthworms. For both soils with 1000 mg of CeO2/kg at 600 °C, biochar significantly decreased the accumulation of Ce in the worm tissues. Amendment with 350 °C biochar had mixed responses on Ce uptake. Analysis by micro X-ray fluorescence (µ-XRF) and micro X-ray absorption near edge structure (µ-XANES) was used to evaluate Ce localization, speciation, and persistence in CeO2- and biochar-exposed earthworms after depuration for 12, 48, and 72 h. Earthworms from the 500 mg of CeO2/kg and 0% biochar treatments eliminated most Ce after a 48 h depuration period. However, in the same treatment and with 5% BC-600 (biochar pyrolysis temperature of 600 °C), ingested biochar fragments (∼50 µm) with Ce adsorbed to the surfaces were retained in the gut after 72 h. Additionally, Ce remained in earthworms from the 2000 mg of CeO2/kg and 5% biochar treatments after depuration for 48 h. Analysis by µ-XANES showed that, within the earthworm tissues, Ce remained predominantly as Ce4+O2, with only few regions (2-3 µm2) where it was found in the reduced form (Ce3+). The present findings highlight that soil and biochar properties have a significant influence in the internalization of CeO2 NPs in earthworms; such interactions need to be considered when estimating NP fate and effects in the environment.

Exposure of agricultural crops to nanoparticle CeO2 in biochar-amended soil.

Biochar is seeing increased usage as an amendment in agricultural soils but the significance of nanoscale interactions between this additive and engineered nanoparticles (ENP) remains unknown. Corn, lettuce, soybean and zucchini were grown for 28 d in two different soils (agricultural, residential) amended with 0-2000 mg engineered nanoparticle (ENP) CeO2 kg-1 and biochar (350 °C or 600 °C) at application rates of 0-5% (w/w). At harvest, plants were analyzed for biomass, Ce content, chlorophyll and lipid peroxidation. Biomass from the four species grown in residential soil varied with species and biochar type. However, biomass in the agricultural soil amended with biochar 600 °C was largely unaffected. Biochar co-exposure had minimal impact on Ce accumulation, with reduced or increased Ce content occurring at the highest (5%) biochar level. Soil-specific and biochar-specific effects on Ce accumulation were observed in the four species. For example, zucchini grown in agricultural soil with 2000 mg CeO2 kg-1 and 350 °C biochar (0.5-5%) accumulated greater Ce than the control. However, for the 600 °C biochar, the opposite effect was evident, with decreased Ce content as biochar increased. A principal component analysis showed that biochar type accounted for 56-99% of the variance in chlorophyll and lipid peroxidation across the plants. SEM and µ-XRF showed Ce association with specific biochar and soil components, while µ-XANES analysis confirmed that after 28 d in soil, the Ce remained largely as CeO2. The current study demonstrates that biochar synthesis conditions significantly impact interactions with ENP, with subsequent effects on particle fate and effects.

Weathering in soil increases nanoparticle CuO bioaccumulation within a terrestrial food chain.

This study evaluates the bioaccumulation of unweathered (U) and weathered (W) CuO in NP, bulk and ionic form (0-400 mg/kg) by lettuce exposed for 70 d in soil co-contaminated with field incurred chlordane. To evaluate CuO trophic transfer, leaves were fed to crickets (Acheta domestica) for 15 d, followed by insect feeding to lizards (Anolis carolinensis). Upon weathering, the root Cu content of the NP treatment increased 214% (327 ± 59.1 mg/kg) over unaged treatment. Cu root content decreased in bulk and ionic treatments from 70-130 mg/kg to 13-26 mg/kg upon aging in soil. Micro X-ray fluorescence (µ-XRF) analysis of W-NP-exposed roots showed a homogenous distribution of Cu (and Ca) in the tissues. Additionally, micro X-ray absorption near-edge (µ-XANES) analysis of W-NP-exposed roots showed near complete transformation of CuO to Cu (I)-sulfur and oxide complexes in the tissues, whereas in unweathered treatment, most root Cu remained as CuO. The expression level of nine genes involved in Cu transport shows that the mechanisms of CuO NPs (and bulk) response/accumulation are different than ionic Cu. The chlordane accumulation by lettuce upon co-exposure to CuO NPs significantly increased upon weathering. Conversely, bulk and ionic exposures decreased pesticide accumulation by plant upon weathering. The Cu cricket fecal content from U-NP-exposed insects was significantly greater than the bulk or ion treatments, suggesting a higher initial NP accumulation followed by significantly greater elimination during depuration. In the lizard, Cu content in the intestine, body and head did not differ as a function of weathering. This study demonstrates that CuO NPs may undergo transformation processes in soil upon weathering that subsequently impact NPs availability in terrestrial food chains.

Molecular Response of Crop Plants to Engineered Nanomaterials.

Functional toxicology has enabled the identification of genes involved in conferring tolerance and sensitivity to engineered nanomaterial (ENM) exposure in the model plant Arabidopsis thaliana (L.) Heynh. Several genes were found to be involved in metabolic functions, stress response, transport, protein synthesis, and DNA repair. Consequently, analysis of physiological parameters, metal content (through ICP-MS quantification), and gene expression (by RT-qPCR) of A. thaliana orthologue genes were performed across different plant species of agronomic interest to highlight putative biomarkers of exposure and effect related to ENMs. This approach led to the identification of molecular markers in Solanum lycopersicum L. and Cucurbita pepo L. (tomato and zucchini) that might not only indicate exposure to ENMs (CuO, CeO2, and La2O3) but also provide mechanistic insight into response to these materials. Through Gene Ontology (GO) analysis, the target genes were mapped in complex interatomic networks representing molecular pathways, cellular components, and biological processes involved in ENM response. The transcriptional response of 38 (out of 204) candidate genes studied varied according to particle type, size, and plant species. Importantly, some of the genes studied showed potential as biomarkers of ENM exposure and effect and may be useful for risk assessment in foods and in the environment.

Carbon Nanomaterials in Agriculture: A Critical Review.

There has been great interest in the use of carbon nano-materials (CNMs) in agriculture. However, the existing literature reveals mixed effects from CNM exposure on plants, ranging from enhanced crop yield to acute cytotoxicity and genetic alteration. These seemingly inconsistent research-outcomes, taken with the current technological limitations for in situ CNM detection, present significant hurdles to the wide scale use of CNMs in agriculture. The objective of this review is to evaluate the current literature, including studies with both positive and negative effects of different CNMs (e.g., carbon nano-tubes, fullerenes, carbon nanoparticles, and carbon nano-horns, among others) on terrestrial plants and associated soil-dwelling microbes. The effects of CNMs on the uptake of various co-contaminants will also be discussed. Last, we highlight critical knowledge gaps, including the need for more soil-based investigations under environmentally relevant conditions. In addition, efforts need to be focused on better understanding of the underlying mechanism of CNM-plant interactions.

Terrestrial Trophic Transfer of Bulk and Nanoparticle La2O3 Does Not Depend on Particle Size.

The bioaccumulation and trophic transfer of bulk and nanoparticle (NP) La2O3 from soil through a terrestrial food chain was determined. To investigate the impact of growth conditions, lettuce (Lactuca sativa) was grown in 350 or 1200 g of bulk/NP amended soil. Leaf tissues were fed to crickets (Acheta domesticus) or darkling beetles (Tenebrionoidea); select crickets were fed to mantises. In the small pot (350 g), La2O3 exposure reduced plant biomass by 23-30% and La tissue content did not differ with particle size. In the large pot (1200 g), biomass was unaffected by exposure and La content in the tissues were significantly greater with bulk particle treatment. Darkling beetles exposed to bulk and NP La2O3-contaminated lettuce contained La at 0.18 and 0.08 mg/kg; respectively (significantly different, P < 0.05). Crickets fed bulk or NP La2O3-exposed lettuce contained 0.53 and 0.33 mg/kg, respectively (significantly different, P < 0.05) with 48 h of depuration. After 7 d of depuration, La content did not differ with particle size, indicating that 48 h may be insufficient to void the digestive system. Mantises that consumed crickets from bulk and NP-exposed treatments contained La at 0.05-0.060 mg/kg (statistically equivalent). These results demonstrate that although La does trophically transfer, biomagnification does not occur and NP levels are equivalent or less than the bulk metal.

CeO2 and ZnO nanoparticles change the nutritional qualities of cucumber (Cucumis sativus).

There is lack of information about the effects of nanoparticles (NPs) on cucumber fruit quality. This study aimed to determine possible impacts on carbohydrates, proteins, mineral nutrients, and antioxidants in the fruit of cucumber plants grown in soil treated with CeO2 and ZnO NPs at 400 and 800 mg/kg. Fourier transform infrared spectroscopy (FTIR) was used to detect changes in functional groups, while ICP-OES and µ-XRF were used to quantify and map the distribution of nutrient elements, respectively. Results showed that none of the ZnO NP concentrations affected sugars; however at 400 mg/kg, CeO2 and ZnO NPs increased starch content. Conversely, CeO2 NPs did not affect starch content but impacted nonreducing sugar content (sucrose). FTIR data showed changes in the fingerprint regions of 1106, 1083, 1153, and 1181, indicating that both NPs altered the carbohydrate pattern. ZnO NPs did not impact protein fractionation; however, CeO2 NPs at 400 mg/kg increased globulin and decreased glutelin. Both CeO2 and ZnO NPs had no impact on flavonoid content, although CeO2 NPs at 800 mg/kg significantly reduced phenolic content. ICP-OES results showed that none of the treatments reduced macronutrients in fruit. In case of micronutrients, all treatments reduced Mo concentration, and at 400 mg/kg, ZnO NPs reduced Cu accumulation. µ-XRF revealed that Cu, Mn, and Zn were mainly accumulated in cucumber seeds. To the best of the authors' knowledge this is the first report on the nutritional quality of cucumber fruit attributed to the impact of CeO2 and ZnO NPs.

Influence of CeO2 and ZnO nanoparticles on cucumber physiological markers and bioaccumulation of Ce and Zn: a life cycle study.

With the dramatic increase in nanotechnologies, it has become increasingly likely that food crops will be exposed to excess engineered nanoparticles (NPs). In this study, cucumber plants were grown to full maturity in soil amended with either CeO2 or ZnO NPs at concentrations of 0, 400, and 800 mg/kg. Chlorophyll and gas exchange were monitored, and physiological markers were recorded. Results showed that, at the concentrations tested, neither CeO2 nor ZnO NPs impacted cucumber plant growth, gas exchange, and chlorophyll content. However, at 800 mg/kg treatment, CeO2 NPs reduced the yield by 31.6% compared to the control (p ≤ 0.07). ICP-MS results showed that the high concentration treatments resulted in the bioaccumulation of Ce and Zn in the fruit (1.27 mg of Ce and 110 mg Zn per kg dry weight). µ-XRF images exhibited Ce in the leaf vein vasculature, suggesting that Ce moves between tissues with water flow during transpiration. To the authors' knowledge, this is the first holistic study focusing on the impacts of CeO2 and ZnO NPs in the life cycle of cucumber plants.

Synchrotron verification of TiO2 accumulation in cucumber fruit: a possible pathway of TiO2 nanoparticle transfer from soil into the food chain.

The transfer of nanoparticles (NPs) into the food chain through edible plants is of great concern. Cucumis sativus L. is a freshly consumed garden vegetable that could be in contact with NPs through biosolids and direct agrichemical application. In this research, cucumber plants were cultivated for 150 days in sandy loam soil treated with 0 to 750 mg TiO2 NPs kg(-1). Fruits were analyzed using synchrotron µ-XRF and µ-XANES, ICP-OES, and biochemical assays. Results showed that catalase in leaves increased (U mg(-1) protein) from 58.8 in control to 78.8 in 750 mg kg(-1) treatment; while ascorbate peroxidase decreased from 21.9 to 14.1 in 500 mg kg(-1) treatment. Moreover, total chlorophyll content in leaves increased in the 750 mg kg(-1) treatment. Compared to control, FTIR spectra of fruit from TiO2 NP treated plants showed significant differences (p ≤ 0.05) in band areas of amide, lignin, and carbohydrates, suggesting macromolecule modification of cucumber fruit. In addition, compared with control, plants treated with 500 mg kg(-1) had 35% more potassium and 34% more phosphorus. For the first time, µ-XRF and µ-XANES showed root-to-fruit translocation of TiO2 in cucumber without biotransformation. This suggests TiO2 could be introduced into the food chain with unknown consequences.

Synchrotron micro-XRF and micro-XANES confirmation of the uptake and translocation of TiO2 nanoparticles in cucumber (Cucumis sativus) plants.

Advances in nanotechnology have raised concerns about possible effects of engineered nanomaterials (ENMs) in the environment, especially in terrestrial plants. In this research, the impacts of TiO(2) nanoparticles (NPs) were evaluated in hydroponically grown cucumber (Cucumis sativus) plants. Seven day old seedlings were treated with TiO(2) NPs at concentrations varying from 0 to 4000 mg L(-1). At harvest, the size of roots and shoots were measured. In addition, micro X- ray fluorescence (micro-XRF) and micro X-ray absorption spectroscopy (micro-XAS), respectively, were used to track the presence and chemical speciation of Ti within plant tissues. Results showed that at all concentrations, TiO(2) significantly increased root length (average >300%). By using micro-XRF it was found that Ti was transported from the roots to the leaf trichomes, suggesting that trichomes are possible sink or excretory system for the Ti. The micro-XANES spectra showed that the absorbed Ti was present as TiO(2) within the cucumber tissues, demonstrating that the TiO(2) NPs were not biotransformed.

Arsenic localization and speciation in the root-soil interface of the desert plant Prosopis juliflora-velutina.

The bioavailability and mobility of arsenic (As) in soils depends on several factors such as pH, organic matter content, speciation, and the concentration of oxides and clay minerals, among others. Plants modify As bioavailability in the rhizosphere; thus, the biogeochemical processes of As in vegetated and non-vegetated soils are different. Changes in As speciation induced by the rhizosphere can be monitored using micro-focused synchrotron-based X-ray fluorescence (µXRF) combined with µX-ray absorption near-edge spectroscopy (µXANES). This research investigated As speciation in the rhizosphere of mesquite (Prosopis juliflora-velutina) plants grown in a sandy clay loam treated with As(III) and As(V) at 40 mg kg(-1). Rhizosphere soil and freeze-dried root tissues of one-month-old plants were analyzed by bulk XAS. Bulk XAS results showed that As(V) was the predominant species in the soil (rhizosphere and non-vegetated), whereas As(III) was dominant in the root tissues from both As(V) and As(III) treated plants. µXAS and µXRF studies of thin sections from resin embedded soil cores revealed the As(III)-S interactions in root tissues and a predominant As-Fe interaction in the soil. This research demonstrated that the combination of bulk XAS and µXAS techniques is a powerful analytical technique for the study of As speciation in soil and plant samples.