Zaira Clemente

Toxicity assessment of TiO2 nanoparticles in zebrafish embryos under different exposure conditions

The popularity of TiO2 nanoparticles (nano-TiO2) lies in their wide range of nanotechnological applications, together with low toxicity. Meanwhile, recent studies have shown that the photocatalytic properties of this material can result in alterations in their behavior in the environment, causing effects that have not yet been fully elucidated. The objective of this study was to evaluate the toxicity of two formulations of nano-TiO2 under different illumination conditions, using an experimental model coherent with the principle of the three Rs of alternative animal experimentation (reduction, refinement, and replacement). Embryos of the fish Danio rerio were exposed for 96h to different concentrations of nano-TiO2 in the form of anatase (TA) or an anatase/rutile mixture (TM), under either visible light or a combination of visible and ultraviolet light (UV). The acute toxicity and sublethal parameters evaluated included survival rates, malformation, hatching, equilibrium, and overall length of the larvae, together with biochemical biomarkers (specific activities of catalase (CAT), glutathione S-transferase (GST), and acid phosphatase (AP)). Both TA and TM caused accelerated hatching of the larvae. Under UV irradiation, there was greater mortality of the larvae of the groups exposed to TM, compared to those exposed to TA. Exposure to TM under UV irradiation altered the equilibrium of the larvae. Alterations in the activities of CAT and GST were indicative of oxidative stress, although no clear dose-response relationship was observed. The effects of nano-TiO2 appeared to depend on both the type of formulation and the illumination condition. The findings contribute to elucidation of the factors involved in the toxicity of these nanoparticles, as well as to the establishment of protocols for risk assessments of nanotechnology.

Activated carbon from pyrolysed sugarcane bagasse: Silver nanoparticle modification and ecotoxicity assessment

Activated carbon from pyrolysed sugarcane bagasse (ACPB) presented pore size ranges from 1.0 to 3.5nm, and surface area between 1200 and 1400m(2)g(-1) that is higher than commonly observed to commercial activated carbon. The ACPB material was successfully loaded with of silver nanoparticles with diameter around 35nm (0.81wt.%). X-ray photoelectron spectroscopy (XPS) analyses showed that the material surface contains metallic/Ag(0) (93.60wt.%) and ionic/Ag(+) states (6.40wt.%). The adsorption capacity of organic model molecules (i.e. methylene blue and phenol) was very efficient to ACPB and ACPB loaded with silver nanoparticles (ACPB-AgNP), indicating that the material modification with silver nanoparticles has not altered its adsorption capacity. ACPB-AgNP inhibited bacteria growth (Escherichia coli), it is a promising advantage for the use of these materials in wastewater treatment and water purification processes. However, ACPB-AgNP showed environmental risks, with toxic effect to the aquatic organism Hydra attenuata (i.e. LC50 value of 1.94mgL(-1)), and it suppressed root development of Lycopersicum esculentum plant (tomato). Finally, this work draw attention for the environmental implications of activated carbon materials modified with silver nanoparticles.

Chitosan nanoparticles loaded the herbicide paraquat: the influence of the aquatic humic substances on the colloidal stability and toxicity.

Polymeric nanoparticles have been developed for several applications, among them as carrier system of pesticides. However, few studies have investigated the fate of these materials in the environment in relation to colloidal stability and toxicity. In nature, humic substances are the main agents responsible for complexation with metals and organic compounds, as well as responsible for the dynamics of these nanoparticles in aquatic and terrestrial environments. In this context, the evaluation of the influence of aquatic humic substances (AHS) on the colloidal stability and toxicity of polymeric nanoparticles of chitosan/tripolyphosphate with or without paraquat was performed. In this study, the nanoparticles were prepared by the ionic gelation method and characterized by size distribution measurements (DLS and NTA), zeta potential, infrared and fluorescence spectroscopy. Allium cepa genotoxicity studies and ecotoxicity assays with the alga Pseudokirchneriella subcapitata were used to investigate the effect of aquatic humic substances (AHS) on the toxicity of this delivery system. No changes were observed in the physical-chemical stability of the nanoparticles due to the presence of AHS using DLS and NTA techniques. However some evidence of interaction between the nanoparticles and AHS was observed by infrared and fluorescence spectroscopies. The ecotoxicity and genotoxicity assays showed that humic substances can decrease the toxic effects of nanoparticles containing paraquat. These results are interesting because they are important for understanding the interaction of these nanostructured carrier systems with species present in aquatic ecosystems such as humic substances, and in this way, opening new perspectives for studies on the dynamics of these carrier systems in the ecosystem.

Coating carbon nanotubes with humic acid using an eco-friendly mechanochemical method: Application for Cu(II) ions removal from water and aquatic ecotoxicity

In this work, industrial grade multi-walled carbon nanotubes (MWCNT) were coated with humic acid (HA) for the first time by means of a milling process, which can be considered an eco-friendly mechanochemical method to prepare materials and composites. The HA-MWCNT hybrid material was characterized by atomic force microscopy (AFM), scanning electron microscopies (SEM and STEM), X-ray photoelectron spectroscopy (XPS), termogravimetric analysis (TGA), and Raman spectroscopy. STEM and AFM images demonstrated that the MWCNTs were efficiently coated by the humic acid, thus leading to an increase of 20% in the oxygen content at the nanotube surface as observed by the XPS data. After the milling process, the carbon nanotubes were shortened as unveiled by SEM images and the values of ID/IG intensity ratio increased due to shortening of the nanotubes and increasing in the number defects at the graphitic structure of carbon nanotubes walls. The analysis of TGA data showed that the quantity of the organic matter of HA on the nanotube surface was 25%. The HA coating was responsible to favor the dispersion of MWCNTs in ultrapure water (i.e. -42mV, zeta-potential value) and to improve their capacity for copper removal. HA-MWCNTs hybrid material adsorbed 2.5 times more Cu(II) ions than oxidized MWCNTs with HNO3, thus evidencing that it is a very efficient adsorbent material for removing copper ions from reconstituted water. The HA-MWCNTs hybrid material did not show acute ecotoxicity to the tested aquatic model organisms (Hydra attenuata, Daphnia magna, and Danio rerio embryos) up to the highest concentration evaluated (10mgL-1). The results allowed concluding that the mechanochemical method is effective to coat carbon nanotubes with humic acid, thus generating a functional hybrid material with low aquatic toxicity and great potential to be applied in environmental nanotechnologies such as the removal of heavy metal ions from water.

Nanoecotoxicity assessment of graphene oxide and its relationship with humic acid

The risk assessment of nanomaterials is essential for regulatory purposes and for sustainable nanotechnological development. Although the application of graphene oxide has been widely exploited, its environmental risk is not well understood because several environmental conditions can affect its behavior and toxicity. In the present study, the graphene oxide effect from aquatic ecosystems was assessed considering the interaction with humic acid on 9 organisms: Raphidocelis subcapitata (green algae), Lemna minor (aquatic plant), Lactuca sativa (lettuce), Daphnia magna (planktonic microcrustacean), Artemia salina (brine shrimp), Chironomus sancticaroli (Chironomidae), Hydra attenuata (freshwater polyp), and Caenorhabditis elegans and Panagrolaimus sp. (nematodes). The no-observed-effect concentration (NOEC) was calculated for each organism. The different criteria used to calculate NOEC values were transformed and plotted as a log-logistic function. The hypothetical 5 to 50% hazardous concentration values were, respectively, 0.023 (0.005-0.056) and 0.10 (0.031-0.31) mg L-1 for graphene oxide with and without humic acid, respectively. The safest scenario associated with the predicted no-effect concentration values for graphene oxide in the aquatic compartment were estimated as 20 to 100 μg L-1 (in the absence of humic acid) and 5 to 23 μg L-1 (in the presence of humic acid). Finally, the present approach contributed to the risk assessment of graphene oxide-based nanomaterials and the establishment of nano-regulations. Environ Toxicol Chem 2018;37:1998-2012.

Toxicity assessment of TiO2-MWCNT nanohybrid material with enhanced photocatalytic activity on Danio rerio (Zebrafish) embryos

The increasing production and use of nanomaterials is causing serious concerns about their safety to human and environmental health. However, the applications of titanium dioxide nanoparticles (TiO2NP) and multiwalled carbon nanotubes (MWCNT) hybrids has grown considerably, due to their enhanced photocatalytic efficiency. To our knowledge, there are no reports available to the scientific community about their toxicity. In this work, we perform a toxicity assessment of TiO2NP and TiO2-MWCNT nanohybrid materials using Zebrafish embryos standardized 96 h early life stage assay, under different exposure conditions (with and without UV light exposure). After exposure the parameters assessed were acute toxicity, hatching rate, growth, yolk sac size, and sarcomere length. In addition, μ-probe X-ray fluorescence spectroscopy (µ-XRF) was employed to observe if nanoparticles were uptaken by zebrafish embryos and consequently accumulated in their organisms. Neither TiO2NP nor TiO2-MWCNT nanohybrids presented acute toxicity to the zebrafish embryos. Moreover, TiO2NP presents sublethal effects for total length (with and without UV light exposure) on the embryos. This work contributes to the understanding of the potential adverse effects of the emerging nanohybrid materials towards safe innovation approaches in nanotechnology.