Magnetic graphene derivates for efficient herbicide removal from aqueous solution through adsorption.

2,4-Dichlorophenoxyacetic acid (2,4-D) is an herbicide and is among the most widely distributed pollutant in the environment and wastewater. Herein is presented a complete comparison of adsorption performance between two different magnetic carbon nanomaterials: graphene oxide (GO) and its reduced form (rGO). Magnetic functionalization was performed employing a coprecipitation method, using only one source of Fe2+, requiring low energy, and potentially allowing the control of the amount of incorporated magnetite. For the first time in literature, a green reduction approach for GO with and without Fe3O4, maintaining the magnetic behavior after the reaction, and an adsorption performance comparison between both carbon nanomaterials are demonstrated. The nanoadsorbents were characterized by FTIR, XRD, Raman, VSM, XPS, and SEM analyses, which demonstrates the successful synthesis of graphene derivate, with different amounts of incorporate magnetite, resulting in distinct magnetization values. The reduction was confirmed by XPS and FTIR techniques. The type of adsorbent reveals that the amount of magnetite on nanomaterial surfaces has significant influence on adsorption capacity and removal efficiency. The procedure demonstrated that the best performance, for magnetic nanocomposites, was obtained by GO∙Fe3O4 1:1 and rGO∙Fe3O4 1:1, presenting values of removal percentage of 70.49 and 91.19%, respectively. The highest adsorption capacity was reached at pH 2.0 for GO∙Fe3O4 1:1 (69.98 mg g-1) and rGO∙Fe3O4 1:1 (89.27 mg g-1), through different interactions: π-π, cation-π, and hydrogen bonds. The adsorption phenomenon exhibited a high dependence on pH, initial concentration of adsorbate, and coexisting ions. Sips and PSO models demonstrate the best adjustment for experimental data, suggesting a heterogeneous surface and different energy sites, respectively. The thermodynamic parameters showed that the process was spontaneous and exothermic. Finally, the nanoadsorbents demonstrated a high efficiency in 2,4-D adsorption even after five adsorption/desorption cycles.

Ecotoxicological assessment of guanitoxin-producing cyanobacteria in Danio rerio and Daphnia similis.

Anthropogenic activity has dramatically deteriorated aquatic ecosystems in recent years. Such environmental alterations could change the primary producers' composition, exacerbating the proliferation of harmful microorganisms such as cyanobacteria. Cyanobacteria can produce several secondary metabolites, including guanitoxin, a potent neurotoxin and the only naturally occurring anticholinesterase organophosphate ever reported in the literature. Therefore, this study investigated the acute toxicity of guanitoxin-producing cyanobacteria Sphaerospermopsis torques-reginae (ITEP-024 strain) aqueous and 50% methanolic extracts in zebrafish (Danio rerio) hepatocytes (ZF-L cell line), zebrafish embryos (fish embryo toxicity - FET) and specimens of the microcrustacean Daphnia similis. For this, hepatocytes were exposed to 1-500 mg/L of the ITEP-024 extracts for 24 h, the embryos to 31.25-500 mg/L for 96 h, and D. similis to 10-3000 mg/L for 48 h. Non-target metabolomics was also performed to analyze secondary metabolites produced by the ITEP-024 using LC-MS/MS. Metabolomics indicated the guanitoxin presence just in the aqueous extract of the ITEP-024 and the presence of the cyanopeptides namalides, spumigins, and anabaenopeptins in the methanolic extract. The aqueous extract decreased the viability of zebrafish hepatocytes (EC(I)50(24h) = 366.46 mg/L), and the methanolic extract was not toxic. FET showed that the aqueous extract (LC50(96) = 353.55 mg/L) was more toxic than the methanolic extract (LC50(96) = 617.91 mg/L). However, the methanolic extract had more sublethal effects, such as abdominal and cardiac (cardiotoxicity) edema and deformation (spinal curvature of the larvae). Both extracts immobilized daphnids at the highest concentration analyzed. However, the aqueous extract was nine times more lethal (EC(I)50(48h) = 108.2 mg/L) than the methanolic extract (EC(I)50(48h) = 980.65 mg/L). Our results showed an imminent biological risk for aquatic fauna living in an ecosystem surrounded by ITEP-024 metabolites. Our findings thus highlight the urgency of understanding the effects of guanitoxin and cyanopeptides in aquatic animals.

Co-exposure of iron oxide nanoparticles with glyphosate herbicides in Poecilia reticulata: Fish liver damages is reversible during iron accumulation and elimination period.

Iron oxide nanoparticles (IONPs) are advanced materials for water remediation technologies. It is therefore relevant to evaluate the cellular and tissue behavior of fishes in response to IONPs and their associations with agrochemicals such as glyphosate (GLY) and glyphosate-based herbicides (GBHs). Iron accumulation, tissue integrity and lipid distribution in the hepatocytes of Poecilia reticulata (guppy) were investigated in a control group and in groups exposed to soluble iron ions, namely IFe (0.3 mgFe/L), IONPs (0.3 mgFe/L), and IONPs, associated with GLY (0.65 mg/L), GBHs 0.65 mgGLY/L (IONPs + GBH1), and 1.30 mgGLY/L (IONPs + GBH2), for 7, 14, and 21 days, followed by an equal period of postexposure in clean reconstituted water. The results showed that the accumulation of iron was greater in the subjects in the IONP treatment group when compared to that in the Ife group. In addition, the subjects in the mixtures with GBHs had a greater accumulation of iron than those in the IONP + GLY treatment group. Tissue integrity assessments demonstrated an intense accumulation of lipids, formation of necrotic zones and leukocyte infiltrates in all the treated groups, with a greater quantity of lipids in the animals treated with IONP + GLY and IFe. During postexposure, the results indicated an elimination of iron in all treated groups, reaching the same level as the control group, throughout the 21 days postexposure. Thus, the damage caused to animal livers by IONP mixtures is reversible, providing promising results for the development of safe environmental remediation practices using nanoparticles.

Toxicity mitigation and biodistribution of albumin corona coated graphene oxide and carbon nanotubes in Caenorhabditis elegans.

In this work, the toxicity and biodistribution of graphene oxide (GO) and oxidized multi-walled carbon nanotubes (MWCNT) were investigated in Caenorhabditis elegans. Bovine serum albumin (BSA) was selected as a model protein to evaluate the influence of protein corona formation on materials physicochemical properties, colloidal stability, and toxicity. Biological assays were performed to assess the effects of bare and albumin corona coated materials on survival, oxidative stress, intestinal barrier permeability, growth, reproduction, and fertility. Critical alterations in topography, surface roughness and chemistry of GO and MWCNT were observed due to albumin corona formation. These modifications were associated with changes in colloidal stability of materials and prevention of their aggregation and sedimentation in nematode testing medium. Both GO and MWCNT caused damage to nematode survival, growth, reproduction, and fertility, as well as enhanced oxidative stress and permeability of the intestinal barrier. But GO was more toxic than MWCNT to C. elegans, especially at long-term assays. Albumin corona mitigated 100% of acute and chronic effects of MWCNT. In contrast, the negative effects of GO were not completely mitigated; GO inhibited 16.2% of nematode growth, 86.5% of reproduction, and 32.0% of fertility at the highest concentration evaluated (10 mg L-1), while corona coated GO mitigated 50% and 100% of fertility and growth, respectively. Confocal Raman spectroscopy imaging was crucial to point out that bare and albumin corona coated GO and MWCNT crossed the C. elegans intestinal barrier reaching its reproductive organs. However, BSA corona protected the nematodes targeted organs from negative effects from MWCNT and blocked its translocation to other tissues, while coated GO was translocated inside the nematode affecting the functionality of crucial organs. In addition, coated MWCNT was excreted after 2 h of food resumption, whereas coated GO still accumulated in the nematode intestine. Our results demonstrate that the materials different translocation and excretion patterns in C. elegans had a relation to the impaired physiological functions of primary and secondary organs. This work is a contribution towards a better understanding of the impacts of protein corona on the toxicity of graphene oxide and carbon nanotubes; essential information for biological applications and nanosafety.

Distribution and behavior of lipid droplets in hepatic cells analyzed by variations of citochemical technique and scanning electron microscopy.

Toxicity evaluations involve the analysis of multiple biomarkers. In this study, the liver, target organ analyzed by treatments with iron concentrations, indicated the accumulation of lipids as a response. Considering that the distribution of lipids in an organ is directly related to the induction of inflammatory processes by aquatic contaminants, this study proposes to carry out an integrative investigation of the behavior and the distribution of lipids in the liver tissue. Techniques of light and electron microscopy were performed in order to propose a new way of assessing and quantifying the distribution of lipid droplets, also presenting methodological alternatives that can be chosen by the reader according to the interests and resources available. Thus, it is assumed that the method begins with the fixation of the liver with Glutaraldehyde 2,5% in PBS 0,1 M and continues with post fixation with osmium tretoxide 1%, which marks lipids. For this proposition, two inclusion methodologies were performed to histological analyses in Historesin and ultrastructural analyses in EMBeed 812. For light microscopy (LM) analyses, cuts were obtained with 2,5 micrometers thickness, which were stained with (1) Mayers hematoxylin and (2) toluidine blue. The images obtained were processed in software Image J Fiji to evidence the lipid distribution in liver.•Cytological reactions with osmium tetroxide constitute low complexity methods that allow the optimization of the localization, identification and quantification of lipid droplets in the liver tissue when analyzed under the conventional light microscope.•Samples included in EMBeed 812 resin commonly used in Transmission Electron Microscopy can be analyzed by SEM-BEC, as complementary analyses for the detection of lipids.•Using SEM-BEC and conventional light microscopy, it is possible to quantify the area occupied by lipid droplets using Image J Fiji software, as these are contrasted due to the reaction with osmium tetroxide.

Hybrid magneto-luminescent iron oxide nanocubes functionalized with europium complexes: synthesis, hemolytic properties and protein corona formation.

The use of hybrid nanostructures based on magneto-luminescent properties is a promising strategy for nano-bio applications and theranostics platforms. In this work, we carried out the synthesis and functionalization of iron oxide nanocubes (IONCs) to obtain multifunctional hybrid nanostructures towards biomedical applications. The IONCs were functionalized with tetraethylorthosilicate, thenoyltrifluoroacetone-propyl-triethoxysilane and europium(iii)-dibenzoylmethane complexes to obtain the materials termed as IOCNCs@SiO2, IONCs@SiO2TTA, IONCs@SiO2TTA-Eu and IONCs@SiO2-TTA-Eu-DBM, respectively. Then, the biological interactions of these nanostructures with red blood cells - RBCs (hemolysis) and human blood plasma (protein corona formation) were evaluated. The XPS spectrocopy and EDS chemical mapping analysis showed that each domain is homogeneously occupied in the hybrid material, with the magnetic core at the center and the luminescent domain on the surface of the hybrid nanomaterial with a core@shell like structure. Futhermore, after each functionalization step, the nanomaterial surface charge drastically changed, with critical impact on RBC lysis and corona formation. While IONCs@SiO2 and IONCs@SiO2-TTA-Eu-DBM showed hemolytic properties in a dose-dependent manner, the IONCs@SiO2TTA-Eu did not present any hemolytic effect up to 300 µg mL-1. Protein corona results showed a pattern of selective adsorption of proteins with each surface of the synthesized hybrid materials. However, as a general result, a suppression of hemolysis after protein corona formation in all tests was verified. Finally, this study provides a solid background for further applications of these hybrid magneto-luminescent materials containing new surface functionalities in the emerging field of medical nanobiotechnology.

Exploring the mechanisms of graphene oxide behavioral and morphological changes in zebrafish.

The presence of natural organic matter such as humic acid (HA) can influence the behavior of graphene oxide (GO) in the aquatic environment. In this study, zebrafish embryos were analyzed after 5 and 7 days of exposure to GO (100 mg L-1) and HA (20 mg L-1) alone or together. The results indicated that, regardless of the presence of HA, larvae exposed to GO for 5 days showed an increase in locomotor activity, reduction in the yolk sac size, and total length and inhibition of AChE activity, but there was no difference in enzyme expression. The statistical analysis indicated that the reductions in total larval length, yolk sac size, and AChE activity in larvae exposed to GO persisted in relation to the control group, but there was a recovery of these parameters in groups also exposed to HA. Larvae exposed to GO for 7 days did not show significant differences in locomotor activity, but the RT-PCR gene expression analysis evidenced an increase in the AChE expression. Since the embryos exposed to GO showed a reduction in overall length, they were submitted to confocal microscopy and their muscle tissue configuration investigated. No changes were observed in the muscle tissue. The results indicated that HA is associated with the toxicity risk modulation by GO and that some compensatory homeostasis mechanisms may be involved in the developmental effects observed in zebrafish.

Effects of multiwalled carbon nanotubes and carbofuran on metabolism in Astyanax ribeirae, a native species.

The study of the toxic effect of carbofuran and multiwalled carbon nanotubes (MWCNTs) on Astyanax ribeirae metabolism is of paramount importance due to the increasing use of this pesticide in agriculture and in the production of nanotubes within the material industry. This study aimed to evaluate the effects of carbofuran, MWCNT, and the combination of these compounds on specific oxygen consumption and excretion of ammonia in A. ribeirae. Therefore, 65 fish were divided into three groups of treatments at varying concentrations: carbofuran (0.01, 0.05, 0.1, and 0.5 mg/L), MWCNT (0.1, 0.25, 0.5, and 1.0 mg/L), and 0.5 mg/L of MWCNT added to carbofuran concentrations (0.01, 0.05, 0.1, and 0.5 mg/L). The average specific oxygen consumption in the groups exposed to carbofuran, compared to the control, increased 73.49% at the 0.01 mg/L concentration and decreased 63.86% and 91.57% with treatments of 0.1 and 0.5 mg/L, respectively. For groups exposed to the MWCNT, there was an 83.91% drop with the 1.0 mg/L treatment, and the carbofuran + MWCNT groups recorded a decrease of 71.09%, 92.77%, and 93.98% at concentrations of 0.05, 0.1, and 0.5 mg/L, respectively. In relation to specific ammonia excretion, in groups exposed to carbofuran compared to the control, there was an increase of 134.37% and 200% with the 0.1 and 0.5 mg/L treatments, respectively. The group exposed to carbofuran + MWCNT experienced a decrease of 60% and 80% with treatments of 0.1 mg/L carbofuran + 0.5 mg/L MWCNT and 0.5 mg/L carbofuran + 0.5 mg/L MWCNT, respectively. Therefore, it was concluded that carbofuran + MWCNT interact, increasing the effects in Astyanax sp.

The Positive Fate of Biochar Addition to Soil in the Degradation of PHBV-Silver Nanoparticle Composites.

The environmental contamination of soils by polymeric and nanomaterials is an increasing global concern. Polymeric composites containing silver nanoparticles (AgNP) are collectively one of the most important products of nanotechnology due to their remarkable antimicrobial activity. Biochars are a promising resource for environmental technologies for remediation of soils considering their high inorganic and organic pollutant adsorption capacity and microbial soil consortium stimulation. In this work we report, for the first time, the use of biochar material as a tool to accelerate the degradation of polyhydroxybutyrate- co-valerate (PHBV) and PHBV composites containing AgNP in a tropical soil system, under laboratory conditions. This positive effect is associated with microbial community improvement, which increased the degradation rate of the polymeric materials, as confirmed by integrated techniques for advanced materials characterization. The addition of 5-10% of sugarcane bagasse biochar into soil has increased the degradation of these polymeric materials 2 to 3 times after 30 days of soil incubation. However, the presence of silver nanoparticles in the PHBV significantly reduced the degradability potential of this nanocomposite by the soil microbial community. These results provide evidence that AgNP or Ag+ ions caused a decline in the total number of bacteria and fungi, which diminished the polymer degradation rate in soil. Finally, this work highlights the great potential of biochar resources for application in soil remediation technologies, such as polymeric (nano)material biodegradation.

Antimicrobial activity of Melaleuca alternifolia nanoparticles in polymicrobial biofilm in situ.

Microbial biofilms represent a challenge in the treatment of infections, due to the low efficacy of the antimicrobials. This study evaluated the antimicrobial effect of nanoparticles of Melaleuca alternifolia (TTO) in dental biofilm. Thirty-eight volunteers used an oral device in situ in situ including four bovine enamel specimens for 07 days. From the fifth day four solutions were applied randomly for each specimen: Physiological Saline Solution (0.85% NaCl) (C+), Chlorhexidine 0.12% (CHX), M. alternifolia oil 0.3% (TTO), and a nanoparticle solution of 0.3% M. alternifolia oil (NPTTO). The nanoparticles of TTO were characterized for pH, IPD, medium size, zeta potential and Transmission Electron Microscopy. Antimicrobial activity was evaluated by viable microorganisms count and the structure of the biofilm by atomic force microscopy. The NPTTO presented pH 6.4, particle diameter of 197.9 ± 1 nm, polydispersion index of 0.242 ± 0.005, zeta potential of -7.12 mV and ±0:27 spherical shape. The C+ resulted in 100% of bacterial vitality, while CHX, TTO and NPTTO showed 34.2%, 51.4% and 25.8%, respectively. The AFM images showed biofilms with an average roughness of 350 nm for C+, 275 nm for CHX, 500 nm for TTO and 100 nm for NPTTO. The NPTTO demonstrated excellent antimicrobial activity in the biofilm formed in situ and will possibly be used in future for the treatment/prevention of oral biofilms.

Histopathological alterations in the gills of Nile tilapia exposed to carbofuran and multiwalled carbon nanotubes.

Carbofuran is a nematicide insecticide with a broad spectrum of action. Carbofuran has noxious effects in several species and has been banned in the USA and Europe; however, it is still used in Brazil. Aquatic organisms are not only exposed to pesticides but also to manufactured nanoparticles, and the potential interaction of these compounds therefore requires investigation. The aim of this study was to examine the histopathological alterations in the gills of Nile tilapia (Oreochromis niloticus) to determine possible effects of exposure to carbofuran, nitric acid-treated multiwalled carbon nanotubes (HNO3-MWCNTs) and the combination of carbofuran with nanotubes. Juvenile fish were exposed to different concentrations of carbofuran (0.1, 0.5, 2.0, 4.0 and 8.0mg/L), different concentrations of HNO3-MWCNTs (0.5, 1.0 and 2.0mg/L) or different concentrations of carbofuran (0.1, 0.5, 2.0, 4.0 and 8.0mg/L) with 1.0mg/L of HNO3-MWCNTs. After 24h of exposure, the animals were removed from the aquarium, the spinal cord was transversely sectioned, and the second gill arch was removed for histological evaluation. Common histological changes included dislocation of the epithelial cells, hyperplasia of the epithelial cells along the secondary lamellae, aneurism, and dilation and disarrangement of the capillaries. All the groups exposed to carbofuran demonstrated a dose-dependent correlation in the Histological Alteration Index; the values found for carbofuran and carbon nanotubes were up to 25% greater than for carbofuran alone. This result indicates an interaction between these toxicants, with enhanced ecotoxic effects. This work contributes to the understanding of the environmental impacts of nanomaterials on aquatic organisms, which is necessary for the sustainable development of nanotechnologies.

Anti-adhesion and antibacterial activity of silver nanoparticles supported on graphene oxide sheets.

This work reports on the preparation, characterization and antibacterial activity of a nanocomposite formed from graphene oxide (GO) sheets decorated with silver nanoparticles (GO-Ag). The GO-Ag nanocomposite was prepared in the presence of AgNO3 and sodium citrate. The physicochemical characterization was performed by UV-vis spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), Raman spectroscopy and transmission electron microscopy (TEM). The average size of the silver nanoparticles anchored on the GO surface was 7.5 nm. Oxidation debris fragments (a byproduct adsorbed on the GO surface) were found to be crucial for the nucleation and growth of the silver nanoparticles. The antibacterial activity of the GO and GO-Ag nanocomposite against the microorganism Pseudomonas aeruginosa was investigated using the standard counting plate methodology. The GO dispersion showed no antibacterial activity against P. aeruginosa over the concentration range investigated. On the other hand, the GO-Ag nanocomposite displayed high biocidal activity with a minimum inhibitory concentration ranging from 2.5 to 5.0 µg/mL. The anti-biofilm activity toward P. aeruginosa adhered on stainless steel surfaces was also investigated. The results showed a 100% inhibition rate of the adhered cells after exposure to the GO-Ag nanocomposite for one hour. To the best of our knowledge, this work provides the first direct evidence that GO-Ag nanocomposites can inhibit the growth of microbial adhered cells, thus preventing the process of biofilm formation. These promising results support the idea that GO-Ag nanocomposites may be applied as antibacterial coatings material to prevent the development of biofilms in food packaging and medical devices.

Inflammatory and hyperalgesic effects of oxidized multi-walled carbon nanotubes in rats.

We evaluated local inflammatory activity of oxidized multiwalled carbon nanotubes in rat experimental models of acute inflammation (paw edema and hyperalgesia) by analyzing their toxicity in non-mesoendothelial tissues. Subcutaneous injection of the nanotubes induced paw edema, that was maximal in the first 2 h after administration at 0.1 mg/kg (43.25 +/- 3.8 AUC) and 1 mg/kg (30.1 +/- 1.8 AUC) compared to saline (18.32 +/- 02.05 AUC). The histopathological analysis showed acute inflammation characterized by vasodilatation, edema formation, neutrophil infiltrate and tissue damage. The nanotubes also elicited hyperalgesic response, seen by the increase of animal paw withdrawal that was maximal in the first 3 hours. The data obtained at the 3rd h was: 75 +/- 9.3% (0.01 mg/kg), 58 +/- 8.3% (0.1 mg/kg) and 53 +/- 6.69% (1 mg/kg) in relation with saline (28 +/- 3.5%). In conclusion, the oxidized multiwalled carbon nanotubes elicit inflammatory and hyperalgesic effects associated to severe tissue damage in rats.

Insecticidal effect of labramin, a lectin-like protein isolated from seeds of the beach apricot tree, Labramia bojeri, on the Mediterranean flour moth, Ephestia kuehniella.

The objective of this work was to study the insecticidal effect of labramin, a protein that shows lectin-like properties. Labramin was isolated from seeds of the Beach Apricot tree, Labramia bojeri A. DC ex Dubard (Ericales: Sapotaceae), and assessed against the development of the Mediterranean flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae), an important pest of stored products such as corn, wheat, rice, and flour. Results showed that labramin caused 90% larval mortality when incorporated in an artificial diet at a level of 1% (w/w). The presence of 0.25% labramin in the diet affected the larval and pupal developmental periods and the percentage of emerging adults. Treatments resulted in elevated levels of trypsin activity in midgut and fecal materials, indicating that labramin may have affected enzyme-regulatory mechanisms by perturbing peritrophic membranes in the midgut of is. kuehniella larvae. The results of dietary experiments with E. kuehniella larvae showed a reduced efficiency for the conversion of ingested and digested food, and an increase in approximate digestibility and metabolic cost. These findings suggest that labramin may hold promise as a control agent to engineer crop plants for insect resistance.