Biogenic synthesis of silver nanoparticles employing fungi offers advantages, including the formation of a capping from fungal biomolecules, which provides stability and can contribute to biological activity. In this work, silver nanoparticles were synthesized using Trichoderma harzianum cultivated with (AgNP-TS) and without enzymatic stimulation (AgNP-T) by the cell wall of Sclerotinia sclerotiorum. The nanoparticles were evaluated for the control of S. sclerotiorum. The specific activity of the T. harzianum hydrolytic enzymes were determined in the filtrates and nanoparticles. Cytotoxicity and genotoxicity were also evaluated. Both the nanoparticles exhibited inhibitory activity towards S. sclerotiorum, with no new sclerotia development, however AgNP-TS was more effective against mycelial growth. Both the filtrates and the nanoparticles showed specific enzymatic activity. Low levels of cytotoxicity and genotoxicity were observed. This study opens perspectives for further exploration of fungal biogenic nanoparticles, indicating their use for the control of S. sclerotiorum and other agricultural pests.
Background Biogenic nanoparticles possess a capping of biomolecules derived from the organism employed in the synthesis, which contributes to their stability and biological activity. These nanoparticles have been highlighted for the control of phytopathogens, so there is a need to understand their composition, mechanisms of action, and toxicity. This study aimed to investigate the importance of the capping and compare the effects of capped and uncapped biogenic silver nanoparticles synthesized using the filtrate of Trichoderma harzianum against the phytopathogenic fungus Sclerotinia sclerotiorum. Capping removal, investigation of the composition of the capping and physico-chemical characterization of the capped and uncapped nanoparticles were performed. The effects of the nanoparticles on S. sclerotiorum were evaluated in vitro. Cytotoxicity and genotoxicity of the nanoparticles on different cell lines and its effects on nontarget microorganisms were also investigated. Results The capped and uncapped nanoparticles showed spherical morphology, with greater diameter of the uncapped ones. Functional groups of biomolecules, protein bands and the hydrolytic enzymes NAGase, β-1,3-glucanase, chitinase and acid protease from T. harzianum were detected in the capping. The capped nanoparticles showed great inhibitory potential against S. sclerotiorum, while the uncapped nanoparticles were ineffective. There was no difference in cytotoxicity comparing capped and uncapped nanoparticles, however higher genotoxicity of the uncapped nanoparticles was observed towards the cell lines. Regarding the effects on nontarget microorganisms, in the minimal inhibitory concentration assay only the capped nanoparticles inhibited microorganisms of agricultural importance, while in the molecular analysis of the soil microbiota there were major changes in the soils exposed to the uncapped nanoparticles. Conclusions The results suggest that the capping played an important role in controlling nanoparticle size and contributed to the biological activity of the nanoparticles against S. sclerotiorum. This study opens perspectives for investigations concerning the application of these nanoparticles for the control of phytopathogens.
BACKGROUND: Botanical compounds from plant species are known to have pesticidal activity and have been used in integrated pest management programs. The varied spectrum of the pesticidal action of these compounds can also avoid selection of resistance in pest populations. In this study, mixtures of the botanical compounds geraniol, eugenol and cinnamaldehyde were encapsulated in zein nanoparticles to improve their stability and efficiency. Biological effects of the nano-scale formulations of the botanical compounds were evaluated against two agricultural pests: the two-spotted spider mite (Tetranychus urticae) and the soybean looper (Chrysodeixis includes).RESULTS: The formulations were stable over time (120 days) with a high encapsulation efficiency (>90%). Nanoencapsulation also provided protection against degradation of the compounds during storage and led to a decrease in toxicity to non-target organisms. The release of the compounds (especially eugenol and cinnamaldehyde) from the nanoparticles was directly influenced by temperature, and the main mechanism of release was through a diffusion-based process. Nanoencapsulated compounds also showed superior efficiency compared to the emulsified compounds in terms of repellency and insecticidal activity. CONCLUSION:The findings of this study indicate that the convergence of botanical compounds with nano-scale formulation has the potential to improve efficacy for their sustainable use in integrated pest management in agriculture.
The biogenic synthesis of metallic nanoparticles can contribute to resolving problems related to pests and soil fertilization. Among the different types of metallic nanoparticles, iron nanoparticles have shown good results, especially concerning toxicity because this metal is an essential micronutrient for all plants and can assist their growth, increasing the levels of carbohydrates, proteins, and chlorophyll. This work performed the green synthesis of biogenic iron oxide nanoparticles using the biological control agent Trichoderma harzianum as a stabilizing agent. The physicochemical properties of the nanoparticles were evaluated using the following techniques: dynamic light scattering, nanoparticle tracking analysis, scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. Cytotoxicity was evaluated using different cell lines, while comet and Allium cepa assays were used to assess genotoxicity. In addition, as a proof of concept, the biological activity of the nanoparticles against the pathogen Sclerotinia sclerotiorum (white mold) was evaluated using an in vitro antifungal activity test. The effect of the nanoparticles on seed germination was also evaluated. The results indicated that the nanoparticles consisted of hematite (α-Fe2O3) and had a mean size diameter of 207 ± 2 nm, polydispersity index of 0.45 ± 0.07, and zeta potential of 13 ± 2 mV. The biogenic iron oxide nanoparticles did not alter cell viability, compared to the controls, and did not lead to changes in the mitotic index, at the concentrations used. Furthermore, they were able to increase the proliferation of Trichoderma, which led to the inhibition of emergence of the pathogen S. sclerotiorum and did not affect the germination of the seeds. Therefore, the green synthesis of biogenic iron oxide nanoparticles based on T. harzianum is an attractive option for pest control, aiming at sustainable agricultural practices.
Silver nanoparticles (AgNPs) are known mainly because of their bactericidal properties. Among the different types of synthesis, there is the biogenic synthesis, which allows the synergy between the nanocomposites and substances from the organism employed for the synthesis. This study describes the synthesis of AgNPs using infusion of roots (AgNpR) and extract (AgNpE) of the plant Althaea officinalis. After the synthesis through reduction of silver nitrate with compounds of A. officinalis, physico-chemical analyzes were performed by UV-Vis spectroscopy, nanoparticles tracking analysis (NTA), dynamic light scattering (DLS) and scanning electron microscopy (SEM). Toxicity was evaluated through Allium cepa assay, comet test with cell lines, cell viability by mitochondrial activity and image cytometry and minimal inhibitory concentration on pathogenic microorganisms. Biochemical analyzes (CAT - catalase, GPx - glutathione peroxidase e GST - glutationa S-transferase) and genotoxicity evaluation in vivo on Zebrafish were also performed. AgNpE and AgNpR showed size of 157 ± 11 nm and 293 ± 12 nm, polydispersity of 0.47 ± 0.08 and 0.25 ± 0.01, and zeta potential of 20.4 ± 1.4 and 26.5 ± 1.2 mV, respectively. With regard to toxicity, the AgNpE were the most toxic when compared with AgNpR. Biochemical analyzes on fish showed increase of CAT activity in most of the organs, whereas GPx showed few changes and the activity of GST decreased. Also regarding to bactericidal activity, both nanoparticles were effective, however AgNpR showed greater activity. Althaea officinalis can be employed as reducing agent for the synthesis of silver nanoparticles, although it is necessary to consider its potential toxicity and ecotoxicity.
Repellents are among the leading products used against diseases transmitted by the Aedes aegypti mosquito. However, their indiscriminate use or high concentrations can cause severe adverse reactions, particularly in children and pregnant women. To protect them, nanotechnology is a promising tool to encapsulate active compounds against degradation, increase their effectiveness, and decrease their toxicity, as it can promote the modified release of the active compound. This study aimed to develop polymeric nanocapsules containing the repellent actives geraniol and icaridin using low concentrations of the active component, with the objective of promoting effective activity and greater safety against adverse reactions. The nanocapsules were developed by the interfacial deposition method, and the physicochemical properties of the nanocapsules were evaluated using dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), zeta potential, differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), release kinetics assay, and mathematical modeling. Cell viability was assessed by the MTT assay and genotoxicity analysis using the comet assay. The developed nanocapsules containing geraniol and icaridin showed mean diameters of 260 nm and 314 nm, respectively, with a polydispersity index < 0.2. The nanocapsules showed encapsulation efficiency values of 73.7 ± 0.1% for icaridin and 98.7 ± 0.1% for geraniol. Morphological analysis showed spherical nanocapsules with low polydispersity. The kinetic parameters calculated using the Korsmeyer–Peppas model indicated an anomalous release profile. Cell viability and genotoxicity analyses showed that the nanocapsules did not alter cell viability or damage DNA. The results demonstrate a promising nanostructured system with good physicochemical characteristics and good stability, with repellent activity against Aedes aegypti.
Dengue, yellow fever, Chinkungunya, Zika virus, and West Nile fever have infected millions and killed a considerable number of humans since their emergence. These arboviruses are transmitted by mosquito bites and topical chemical repellents are the most commonly used method to protect against vector arthropod species. This study aimed to develop a new generation of repellent formulations to promote improved arboviruses transmission control. A repellent system based on polycaprolactone (PCL)-polymeric nanoparticles was developed for the dual encapsulation of IR3535 and geraniol and further incorporation into a thermosensitive hydrogel. The physicochemical and morphological parameters of the prepared formulations were evaluated by dynamic light scattering (DLS), nano tracking analysis (NTA), atomic force microscopy (AFM). In vitro release mechanisms and permeation performance were evaluated before and after nanoparticles incorporation into the hydrogels. FTIR analysis was performed to evaluate the effect of formulation epidermal contact. Potential cytotoxicity was evaluated using the MTT reduction test and disc diffusion methods. The nanoparticle formulations were stable over 120 days with encapsulation efficiency (EE) of 60% and 99% for IR3535 and geraniol, respectively. AFM analysis revealed a spherical nanoparticle morphology. After 24 h, 7 ± 0.1% and 83 ± 2% of the GRL and IR3535, respectively, were released while the same formulation incorporated in poloxamer 407 hydrogel released 11 ± 0.9% and 29 ± 3% of the loaded GRL and IR3535, respectively. GRL permeation from PCL nanoparticles and PCL nanoparticles in the hydrogel showed similar profiles, while IR3535 permeation was modulated by formulation compositions. Differences in IR3535 permeated amounts were higher for PCL nanoparticles in the hydrogels (36.9 ± 1.1 mg/cm2) compared to the IR3535-PCL nanoparticles (29.2 ± 1.5 mg/cm2). However, both active permeation concentrations were low at 24 h, indicating that the formulations (PCL nanoparticles and PCL in hydrogel) controlled the bioactive percutaneous absorption. Minor changes in the stratum corneum (SC) caused by interaction with the formulations may not represent a consumer safety risk. The cytotoxicity results presented herein indicate the carrier systems based on poly-epsilon caprolactone (PCL) exhibited a reduced toxic effect when compared to emulsions, opening perspectives for these systems to be used as a tool to prolong protection times with lower active repellent concentrations.
EFFECTS OF COMMERCIAL NANOPARTICLES OF IRON OXIDE (Fe 2 O 3 ): CYTOTOXICITY, GENOTOXICITY AND OXIDATIVE STRESS. This study presents the toxicological effects of iron nanoparticles (NP Fe 2 O 3 ) using in vitro and in vivo tests. Initially nanoparticles were characterized physic-chemically followed by evaluation of cell viability using different cell lines. Allium cepa test and comet assay (cell lines and Danio rerio) were used for evaluation of genotoxicity. Oxidative stress analyses were performed using D. rerio exposed to NP-Fe 2 O 3 , and the enzymatic activity of the enzymes catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) were evaluated. The results showed that initially nanoparticles had 65.55 nm, 0.24 of polydispersity of and 11.4 mV of zeta potential. Regarding cell viability, it was observed that this did not reach the IC 50 up to the concentration 1 x 10 10 NP mL -1. The in vitro comet assay showed that in concentrations 1.96 x10 8 and 10 9 NPs mL -1 these presented toxicity, for Allium cepa evaluation in concentrations 19.6 and 39.0 x10 9 NPs mL -1 presented significant damages when compared to the control. Oxidative stress showed that the liver was the most affected organ when compared to the control. Although studies show that iron nanoparticles do not lead to changes, further studies are needed to make sure they do not lead to environmental changes.Keywords: iron oxide nanoparticles; zebrafish; viability; in vitro analysis; in vivo analysis. INTRODUÇÃOCom o avanço da nanotecnologia foi possível produzir nanopartículas de ferro as quais apresentam características físico-químicas diferenciadas quando comparadas às características apresentadas em micro/macroescala. Além disso, o átomo de ferro pode originar outros compostos a partir de diferentes combinações, como por exemplo os óxidos de ferro.1,2 Dentre as nanopartículas (NPs) metálicas, as de óxido de ferro se destacam no mercado devido às suas caracterís-ticas físico-químicas, como sua grande capacidade cinética, grande área de superfície em relação ao volume, alta reatividade, possíveis propriedades magnéticas, além de permitirem o recobrimento com diversos tipos de ligantes específicos. 3-7Atualmente a maioria dos estudos de NPs de óxidos de ferro (NP Fe 2 O 3 ) na área ambiental está direcionada à remediação (nanorremediação), que consiste na utilização das nanoparticulas para a descontaminação da água e solo. 5,8 Em relação a área médica, sua utilização ocorre principalmente como meio de contraste em imagens por ressonância magnética, carreadores de fármacos, em pesquisas gênicas, proteômica e tratamentos anticâncer. 2,6,7,[9][10][11][12] As NP Fe 2 O 3 também são utilizadas como constituintes de diferentes tipos de materiais comercializados: tintas, revestimentos, plásticos, cosméticos, 13 nutrientes 2 e semicondutores. 8 Assim, devido à ampla comercialização de produtos que apresentam NPs em sua composição, é observado como consequência um aumento do descarte das mesmas nos ambientes aquático e terrestre. [14]...
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