Photodynamic inactivation of methicillin-resistant Staphylococcus aureus by using Giemsa dye as a photosensitizer.

The rise of antibiotic-resistant bacteria calls for innovative approaches to combat multidrug-resistant strains. Here, the potential of the standard histological stain, Giemsa, to act as a photosensitizer (PS) for antimicrobial photodynamic inactivation (aPDI) against methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) strains is reported. Bioassays were performed using various Giemsa concentrations (ranging from 0.0 to 20.0 µM) under 625 nm illumination at a light dose of 30 J cm-2. Remarkably, Giemsa completely inhibited the growth of MSSA and MRSA bacterial colonies for concentrations at 10 µM and higher but exhibited no inhibitory effect without light exposure. Partition coefficient analysis revealed Giemsa's affinity for membranes. Furthermore, we quantified the production of reactive oxygen species (ROS) and singlet oxygen (1O2) to elucidate the aPDI mechanisms underlying bacterial inactivation mediated by Giemsa. These findings highlight Giemsa stain's potential as a PS in aPDI for targeting multidrug-resistant bacteria.

A fluorescence-based multivariate method for biodiesel quantification in undiluted diesel-biodiesel blends without sample preparation.

In this work, excitation-emission matrices (EEMs) were used in association with parallel factor analysis (PARAFAC) to assess biodiesel content in undiluted diesel-biodiesel blends (DBBs) without pre-sample preparation. EEMs were decomposed using the PARAFAC (EEMs-PARAFAC), and the loading values of the PARAFAC component as a function of biodiesel content in the blends were used to build an analytical model to quantify the biodiesel content in DBBs. The proposed model presenting a limit of detection (LOD) and a limit of quantification (LOQ) of 2.5% and 11% w/w, respectively, successfully predicted the biodiesel content in the validation samples. The robustness of the model was confirmed by a close analysis of the root mean square error of prediction, standard error of prediction, relative standard deviation of prediction, and Bias. Therefore, an accurate and robust analytical model based on EEMs-PARAFAC was developed to quantify the biodiesel content in undiluted DBBs without sample preparation.

Assessing the Fate of Superparamagnetic Iron Oxide Nanoparticles Carrying Usnic Acid as Chemical Cargo on the Soil Microbial Community.

In the present study we evaluate the effect of superparamagnetic iron oxide nanoparticles (SPIONs) carrying usnic acid (UA) as chemical cargo on the soil microbial community in a dystrophic red latosol (oxysol). Herein, 500 ppm UA or SPIONs-framework carrying UA were diluted in sterile ultrapure deionized water and applied by hand sprayer on the top of the soil. The experiment was conducted in a growth chamber at 25 °C, with a relative humidity of 80% and a 16 h/8 h light-dark cycle (600 lx light intensity) for 30 days. Sterile ultrapure deionized water was used as the negative control; uncapped and oleic acid (OA) capped SPIONs were also tested to assess their potential effects. Magnetic nanostructures were synthesized by a coprecipitation method and characterized by scanning and transmission electron microscopy (SEM and TEM), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), zeta potential, hydrodynamic diameter, magnetic measurements, and release kinetics of chemical cargo. Uncapped and OA-capped SPIONs did not significantly affect soil microbial community. Our results showed an impairment in the soil microbial community exposed to free UA, leading to a general decrease in negative effects on soil-based parameters when bioactive was loaded into the nanoscale magnetic carrier. Besides, compared to control, the free UA caused a significant decrease in microbial biomass C (39%), on the activity of acid protease (59%), and acid phosphatase (23%) enzymes, respectively. Free UA also reduced eukaryotic 18S rRNA gene abundance, suggesting a major impact on fungi. Our findings indicate that SPIONs as bioherbicide nanocarriers can reduce the negative impacts on soil. Therefore, nanoenabled biocides may improve agricultural productivity, which is important for food security due to the need of increasing food production.

Miniaturized 3D-Printed Cell Enables Water/Ethanol Quantification Using Electrochemical Impedance Spectroscopy.

A miniaturized and low-cost electrochemical 3D-printed system for rapid and accurate quantification of ethanol content in ethanol fuel using electrochemical impedance spectroscopy (EIS) was developed. The monolithic design of the system incorporates insulating thermoplastic electrode separators, with only the cover being mobile, allowing for easy assembly and handling. The portable device, measuring approximately 26 × 24 mm, has a maximum capacity of 1 mL, making it suitable for lab-on-a-chip and portable analysis. By utilizing the dielectric constant of ethanol and ethanol fuel mixtures with water, the miniaturized EIS cell quantifies ethanol content effectively. To validate its performance, we compared measurements from four gas stations with a digital densimeter, and the values obtained from the proposed system matched perfectly. Our miniaturized and low-cost electrochemical 3D-printed device can be printed and assembled in two hours, offering a cost-effective solution for fast and precise ethanol quantification. Its versatility, affordability, and compatibility with lab-on-a-chip platforms make it easily applicable, including for fuel quality control and on-site analysis in remote locations.

Preparation and Phytotoxicity Evaluation of Cellulose Acetate Nanoparticles.

The use of biocompatible and low-cost polymeric matrices to produce non-phytotoxic nanoparticles for delivery systems is a promising alternative for good practices in agriculture management and biotechnological applications. In this context, there is still a lack of studies devoted to producing low-cost polymeric nanoparticles that exhibit non-phytotoxic properties. Among the different polymeric matrices that can be used to produce low-cost nanoparticles, we can highlight the potential application of cellulose acetate, a natural biopolymer with biocompatible and biodegradable properties, which has already been used as fibers, membranes, and films in different agricultural and biotechnological applications. Here, we provided a simple and low-cost route to produce cellulose acetate nanoparticles (CA-NPs), by modified emulsification solvent evaporation technique, with a main diameter of around 200 nm and a spherical and smooth morphology for potential use as agrochemical nanocarriers. The non-phytotoxic properties of the produced cellulose acetate nanoparticles were proved by performing a plant toxic test by Allium cepa assay. The cytotoxicity and genotoxicity tests allowed us to evaluate the mitotic process, chromosomal abnormalities, inhibition/delay in root growth, and micronucleus induction. In summary, the results demonstrated that CA-NPs did not induce phytotoxic, cytotoxic, or genotoxic effects, and they did not promote changes in the root elongation, germination or in the mitotic, chromosomal aberration, and micronucleus indices. Consequently, the present findings indicated that CA-NPs can be potentially used as environmentally friendly nanoparticles.

Oxidative stability and elemental analysis of sunflower (Helianthus annuus) edible oil produced in Brazil using a domestic extraction machine.

The consumption of regular vegetable oils has been linked to energy acquisition, nutritional benefits, health improvement, and the regulation of metabolic diseases. This study evaluated fatty acids composition, physicochemical, thermal, oxidative, and optical properties, and quantified trace elements in the sunflower oil extracted by a domestic cold-press machine. The oil presented linoleic (54.00%) and oleic (37.29%) primary unsaturated fatty acids (91.67%), in which atherogenic (0.05), thrombogenic (0.16), hypocholesterolemic/hypercholesterolemic (21.97), peroxide (16.16), saponification (141.80), and relative density indices (0.92) demonstrated to be suitable for human consumption and possible health promotion. In addition, the concentrations of trace elements by ICP OES were ordered Zn > Fe > Al > Cu > Mn > Cr. Concentrations of Zn, Fe, Al, Cu, and Mn were lower than FAO/WHO and DRI/AI limits, while Cr concentrations exceeded the FAO/WHO limits, which can be used as an indicator of the polluted ambiance. Sunflower oil quantities daily consumption were calculated by taking into account non-carcinogenic risk (CR < 10-4), and total non-carcinogenic hazard index (HI < 1). Based on trace elements determined in this study, the suitable quantity of sunflower oil consumption varies according to individuals aged 8, 18, and 30 years and will be deemed 0.61, 1.46, and 1.65 g/kg, respectively, attending HI = 0.99 and CR < 10-4.

Dissolved organic matter in bovine slaughterhouse wastewater using fluorescence spectroscopy associated with CP/PARAFAC and PCA methods.

In this work, we evaluated the potential application of fluorescence spectroscopy, associated with the canonical polyadic/parallel factor analysis and principal component analysis, to monitor the dissolved organic matter (DOM) generated from a slaughterhouse industry. During the monitoring process, we analyzed the residual water at the entrance and exit sites of the slaughterhouse effluent treatment as well as downstream and upstream the effluent receiving water body of a local river. The results revealed that the fluorescence analysis was able to identify proteins, chlorophylls, and humic substances at the entrance and exit sites of the slaughterhouse treatment plant and humic substances at the river water bodies. Our data also demonstrated that the industrial effluent discharged into the river did not impact the receiving water body quality as determined by the biological and humification indices obtained by fluorescence analysis, which was confirmed by conventional physicochemical analysis. In summary, the present findings indicate that fluorescence spectroscopy, in association with multivariate analysis, can be successfully applied as an analytical tool for evaluating the quality of DOM in slaughterhouse wastewater.

Antimicrobial and Photoantimicrobial Activities of Chitosan/CNPPV Nanocomposites.

Multidrug-resistant bacteria represent a global health and economic burden that urgently calls for new technologies to combat bacterial antimicrobial resistance. Here, we developed novel nanocomposites (NCPs) based on chitosan that display different degrees of acetylation (DAs), and conjugated polymer cyano-substituted poly(p-phenylene vinylene) (CNPPV) as an alternative approach to inactivate Gram-negative (E. coli) and Gram-positive (S. aureus) bacteria. Chitosan's structure was confirmed through FT-Raman spectroscopy. Bactericidal and photobactericidal activities of NCPs were tested under dark and blue-light irradiation conditions, respectively. Hydrodynamic size and aqueous stability were determined by DLS, zeta potential (ZP) and time-domain NMR. TEM micrographs of NCPs were obtained, and their capacity of generating reactive oxygen species (ROS) under blue illumination was also characterized. Meaningful variations on ZP and relaxation time T2 confirmed successful physical attachment of chitosan/CNPPV. All NCPs exhibited a similar and shrunken spherical shape according to TEM. A lower DA is responsible for driving higher bactericidal performance alongside the synergistic effect from CNPPV, lower nanosized distribution profile and higher positive charged surface. ROS production was proportionally found in NCPs with and without CNPPV by decreasing the DA, leading to a remarkable photobactericidal effect under blue-light irradiation. Overall, our findings indicate that chitosan/CNPPV NCPs may constitute a valuable asset for the development of innovative strategies for inactivation and/or photoinactivation of bacteria.

Environmentally Safe Photodynamic Control of Aedes aegypti Using Sunlight-Activated Synthetic Curcumin: Photodegradation, Aquatic Ecotoxicity, and Field Trial.

This study reports curcumin as an efficient photolarvicide against Aedes aegypti larvae under natural light illumination. Larval mortality and pupal formation were monitored daily for 21 days under simulated field conditions. In a sucrose-containing formulation, a lethal time 50 (LT50) of 3 days was found using curcumin at 4.6 mg L-1. This formulation promoted no larval toxicity in the absence of illumination, and sucrose alone did not induce larval phototoxicity. The photodegradation byproducts (intermediates) of curcumin were determined and the photodegradation mechanisms proposed. Intermediates with m/z 194, 278, and 370 were found and characterized using LC-MS. The ecotoxicity of the byproducts on non-target organisms (Daphnia, fish, and green algae) indicates that the intermediates do not exhibit any destructive potential for aquatic organisms. The results of photodegradation and ecotoxicity suggest that curcumin is environmentally safe for non-target organisms and, therefore, can be considered for population control of Ae. aegypti.

Synthesis and Characterization of N and Fe-Doped TiO2 Nanoparticles for 2,4-Dimethylaniline Mineralization.

The present study aimed to evaluate the feasibility of developing low-cost N- and Fe-doped TiO2 photocatalysts for investigating the mineralization of 2,4-dimethylaniline (2,4-DMA). With a single anatase phase, the photocatalysts showed high thermal stability with mass losses of less than 2%. The predominant oxidative state is Ti4+, but there is presence of Ti3+ associated with oxygen vacancies. In materials with N, doping was interstitial in the NH3/NH4+ form and for doping with Fe, there was a presence of Fe-Ti bonds (indicating substitutional occupations). With an improved band gap energy from 3.16 eV to 2.82 eV the photoactivity of the photocatalysts was validated with an 18 W UVA lamp (340-415 nm) with a flux of 8.23 × 10-6 Einstein s-1. With a size of only 14.45 nm and a surface area of 84.73 m2 g-1, the photocatalyst doped with 0.0125% Fe mineralized 92% of the 2,4-DMA in just 180 min. While the 3% N photocatalyst with 12.27 nm had similar performance at only 360 min. Factors such as high surface area, mesoporous structure and improved Ebg, and absence of Fe peak in XPS analysis indicate that doping with 0.0125% Fe caused a modification in TiO2 structure.

Application of Optical Fluorescence Spectroscopy for Studying Bee Abundance in Tropaeolum majus L. (Tropaeolaceae).

Tropaeolum majus L. species produce flowers with all sorts of colors, from yellow to red. This work aimed to apply optical fluorescence spectroscopy to study bee abundance in T. majus, answering the following questions: (1) do corolla temperature and weather conditions affect the abundance of visiting bee species? (2) do flower color and corolla fluorescence affect the abundance of visiting bee species? (3) do red flowers attract more visiting bees? (4) is there a relationship between bee visits and flower compounds? The bee species Apis mellifera, Paratrigona lineata, and Trigona spinipes were the most observed in T. majus flowers. The latter was more active in the morning and preferred orange and yellow flowers. These colors also had higher temperatures and fluorescence emissions than did the red ones and those with yellow-red and orange-red nectar guides. Orange flowers emitted a broadband UV spectrum (between 475 and 800 nm). This range might be due to compounds such as hydroxycinnamic acid, flavonols, isoflavonoids, flavones, phenolic acid, and chlorophyll. Extracts from different T. majus corolla colors showed that flowers emit specific fluorescent signals, mainly related to bee color vision and learning, thus acting as a means of communication between bees and flowers. In this way, this information evidences the interaction between bees and T. majus flowers, allowing conservation actions for pollinators.

Characterization of Buriti (Mauritia flexuosa) Pulp Oil and the Effect of Its Supplementation in an In Vivo Experimental Model.

Mauritia flexuosa (Buriti) pulp oil contains bioactive substances and lipids that are protective against cardiovascular and inflammatory diseases. We performed physical and chemical analyses to verify its quality and stability. Buriti oil was stable according to the Rancimat test, presenting an induction period of 6.6 h. We evaluated the effect of supplementation with crude buriti oil and olive oil on metabolic parameters in 108 Swiss mice for 90 days. We investigated six groups: extra virgin olive oil (EVOO) 1 and 2 (1000 and 2000 mg/kg), buriti oil (BO) 1 and 2 (1000 and 2000 mg/kg), synergic (S) (BO1 + EVOO1), and control (water dose 1000 mg/kg). The animals were euthanized to examine their blood, livers, and fats. The supplementation did not interfere with food consumption, weight gain, and histological alterations in the liver. Group S showed the strongest relationship with the fractions HDL-c and non-HDL-c, indicating a possible cardioprotective effect. Moreover, we observed significantly higher IL-6 levels in the control, EVOO2, and BO1 groups than in the EVOO1 group. Resistin was also significantly higher for the synergic treatment than for the control. We conclude that BO combined with EVOO could be an excellent food supplement for human consumption.

Green synthesis of silver indium telluride nanocrystals: characterization and photothermal analyses.

Here, we report the preparation and characterization of stable AgIn5Te8 (AITe) nanocrystals (NCs) and their respective AITe-ZnS and AITe-ZnSe alloys. A green route of synthesis is described by using an aqueous electrochemical method in a cavity cell. The elemental ratio modulation of the NCs was investigated to determine the ideal improvement of the structural and electronic properties. The AITe NCs presented an emission band at 889 nm and hypsochromic shifts when coated with ZnS and ZnSe seeds. Photothermal analyses point to theranostic applications.

Eco-Friendly Production of AuNPs and Their Impact on the Oil Oxidative Stability.

Vegetable oils have been used for different applications and, more recently, as an active host medium to obtain nanoparticles for employment in bionanotechnological applications. Nevertheless, oils are very susceptible to oxidation during production, storage, and transportation because of their chemical composition. Consequently, any modification in their production must be accompanied by an analysis of the oxidative stability. In this study, naked and biocompatible gold nanoparticles (AuNPs) were grown on sunflower oil during sputtering deposition using different deposition times. Size and morphology were determined by transmission electron microscopy (TEM) and their concentrations were found by inductively coupled plasma-optical emission spectroscopy (ICP-OES). Rancimat® method was employed to evaluate the AuNPs influence on the oxidative stability of the vegetable oil. Well-dispersed quasi-spherical NPs were produced with a mean diameter in the 2.9-3.7 nm range and they were concentration-dependent on the deposition time. A concentration of about 11 mg/L, 38 mg/L, and 225 mg/L of AuNPs was obtained for a deposition time of 5 min, 15 min, and 30 min, respectively. The results also revealed that AuNPs negatively affected the oxidative stability of the sunflower oil and exponentially reduced the induction period (IP) with the increase in AuNPs content. IP reductions of 63%, 77%, and 81% were determined for the AuNPs containing samples at 11 mg/L, 38 mg/L, and 225 mg/L. For the first time, it is reported that naked AuNPs promote the rapid degradation of vegetable oil and this points out the need for attention relative to the quality of vegetable oils used to host metal nanoparticles.

Effects of copper oxide nanoparticles on germination of Sesbania virgata (FABACEAE) plants.

Nanotechnology is a field that, over the years, has been growing in several research areas, such as medicine, agriculture and cosmetics, among others. As a result, there is a continuous increase in the production, use and disposal of these materials in the environment. The behaviour and (bio) activity of these materials in the atmosphere, water and soil are not fully studied. Therefore, it is necessary to carry out an analysis of the risks of contamination, as well as the possible effects and impacts of nanoparticles (NPs) on the ecosystem. In an attempt to investigate these effects on plants, the present study aimed to investigate the impact of copper oxide nanoparticles (CuO NPs) on the seed germination process of Sesbania virgata. For this, the Sesbania virgata seeds were subjected to different concentration of CuO NPs (0, 100, 200, 300 and 400 mgL-1) and their germination and development were monitored by optical analysis (thermography and chlorophyll a fluorescence). The results show that the CuO NPs induced a reduction on the maximum emission of chlorophyll a, which was concentration-dependent. The data also showed that CuO NPs promoted an increase in the energy dissipated by non-photochemical pathways and the surface temperature of the seeds. Additionally, our findings revealed that CuO NPs caused a root growth inhibition. In summary, the present study demonstrates, for the first time, that CuO NPs can negatively affect the physiological status and development of the S. virgata plant, by altering the efficiency of the functioning of photosystem II in its initial developmental stage, depending on the concentration of CuO NPs.

Photodynamic Inactivation of Methicillin-Resistant Staphylococcus aureus by a Natural Food Colorant (E-141ii).

This study evaluates the photosensitizing effectiveness of sodium copper chlorophyllin, a natural green colorant commonly used as a food additive (E-141ii), to inactivate methicillin-sensitive and methicillin-resistant Staphylococcus aureus under red-light illumination. Antimicrobial photodynamic inactivation (aPDI) was tested on a methicillin-sensitive reference strain (ATCC 25923) and a methicillin-resistant Staphylococcus aureus strain (GenBank accession number Mh087437) isolated from a clinical sample. The photoinactivation efficacy was investigated by exposing the bacterial strains to different E-141ii concentrations (0.0, 1.0, 2.5, 5.0, 10.0, and 20.0 µM) and to red light (625 nm) at 30 J cm-2. The results showed that E-141ii itself did not prevent bacterial growth for all tested concentrations when cultures were placed in the dark. By contrast, E-141ii photoinactivated both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) under red-light illumination. However, different dose responses were observed for MSSA and MRSA. Whilst the MSSA growth was inhibited to the detection limit of the method with E-141ii at 2.5 µM, >10 µM concentrations were required to inhibit the growth of MRSA. The data also suggest that E-141ii can produce reactive oxygen species (ROS) via Type I reaction by electron transfer from its first excited singlet state to oxygen molecules. Our findings demonstrate that the tested food colorant has great potential to be used in aPDI of MRSA.

Caryocar brasiliense Cambess. Pulp Oil Supplementation Reduces Total Cholesterol, LDL-c, and Non-HDL-c in Animals.

The fruit of Caryocar brasiliense Cambess. is a source of oil with active compounds that are protective to the organism. In our work, we analyzed the physicochemical characteristics and evaluated the effects of supplementation with C. brasiliense oil in an animal model. We characterized the oil by indices of quality and identity, optical techniques of absorption spectroscopy in the UV-Vis region and fluorescence, and thermogravimetry/derived thermogravimetry (TG/DTG). For the animal experiment, we utilized mice (Mus musculus) supplemented with lipidic source in different dosages. The results demonstrated that C. brasiliense oil is an alternative source for human consumption and presents excellent oxidative stability. Primarily, it exhibited oleic MFA (53.56%) and palmitic SFA (37.78%). The oil level of tocopherols and tocotrienols was superior to the carotenoids. The supplementation with C. brasiliense oil reduced the levels of total cholesterol, LDL-c, and non-HDL-c. Regarding visceral fats and adiposity index, the treatment synergically supplemented with olive oil and C. brasiliense oil (OO + CO) obtained the best result. Therefore, C. brasiliense oil is a high quality product for consumption. Its supplementation promotes beneficial effects mainly on the lipidic profile.

Effective killing of bacteria under blue-light irradiation promoted by green synthesized silver nanoparticles loaded on reduced graphene oxide sheets.

Graphene oxide (GO) materials loaded with silver nanoparticles (AgNPs) have drawn considerable attention due to their capacity to efficiently inactivate bacteria though a multifaceted mechanism of action, as well as for presenting a synergetic effect against bacteria when compared to the activity of AgNPs and GO alone. In this investigation, we present an inexpensive and environmentally-friendly method for synthesizing reduced GO sheets coated with silver nanoparticles (AgNPs/r-GO) using a coffee extract solution as a green reducing agent. The physical and chemical properties of the produced materials were extensively characterized by scanning electron microscopy (SEM), field-emission gun transmission electron microscopy (FEG-TEM), ultraviolet and visible absorption (UV-Vis), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), inductively coupled plasma-optical emission spectroscopy (ICP-OES) and ion release determination. The results demonstrated that AgNPs/r-GO composites were successfully produced, revealing the formation of micrometer-sized r-GO sheets decorated by AgNPs of approximately 70 nm diameter. Finally, bactericidal and photobactericidal effects of the AgNPs/r-GO composites were tested against Staphylococcus aureus, in which the results showed that the composites presented antimicrobial and photoantimicrobial activities. Moreover, our results demonstrated for the first time, to our knowledge, that an efficient process of bacterial inactivation can be achieved by using AgNPs/r-GO composites under blue light irradiation as a result of three different bacterial killing processes: (i) chemical effect promoted by Ag+ ion release from AgNPs; (ii) photocatalytic activity induced by AgNPs/r-GO composites, enhancing the bacterial photoinactivation due to the excited-Plasmons of the AgNPs when anchored on r-GO; and (iii) photodynamic effect produced by bacterial endogenous photosensitizers under blue-light irradiation. In summary, the present findings demonstrated that AgNPs/r-GO can be obtained by a non-toxic procedure with great potential for biomedical-related applications.

In vitro and in vivo impact assessment of eco-designed CuO nanoparticles on non-target aquatic photoautotrophic organisms.

This work has assessed the impact of copper oxide nanoparticles (CuONPs), designed via green route, toward photosynthetic apparatus on aquatic photoautotrophic organisms. In order to filling knowledge gaps, in vitro and in vivo assays were performed, using cyanobacterial phycocyanin (C-PC) from Arthrospira platensis and Lemna valdiviana plants (duckweed), respectively. Impairment in light energy transfer became evident in C-PC exposed to CuONPs, giving rise to an increase of light absorption and a suppression of fluorescence emission. Fourier transform infrared spectroscopy (FTIR) results showed that C-PC structures might be altered by the nanoparticles, also revealed that CuONPs preferably interacts with -NH functional groups. The data also revealed that CuONPs affected the chlorophyll a content in duckweed leaves. In addition, photosystem II (PSII) performance was significantly affected by CuONPs, negatively impacting the PSII photochemical network. In summary, the results point out that, even eco-friendly designed, CuONPs may negatively affect the photosynthetic process when accumulated by aquatic photoautotrophs.

Phytotoxicity of silver nanoparticles on Vicia faba: Evaluation of particle size effects on photosynthetic performance and leaf gas exchange.

Nanotechnology is an emerging field in science and engineering, which presents significant impacts on the economy, society and the environment. The nanomaterials' (NMs) production, use, and disposal is inevitably leading to their release into the environment where there are uncertainties about its fate, behaviour, and toxicity. Recent works have demonstrated that NMs can penetrate, translocate, and accumulate in plants. However, studies about the effects of the NMs on plants are still limited because most investigations are carried out in the initial stage of plant development. The present study aimed to evaluate and characterize the photochemical efficiency of photosystem II (PSII) of broad bean (Vicia faba) leaves when subjected to silver nanoparticles (AgNPs) with diameters of 20, 51, and 73 nm as well as to micrometer-size Ag particles (AgBulk). The AgNPs were characterized by transmission electron microscopy and dynamic light scattering. The analyses were performed by injecting the leaves with 100 mg L-1 aqueous solution of Ag and measuring the chlorophyll fluorescence imaging, gas exchange, thermal imaging, and reactive oxygen species (ROS) production. In addition, silver ion (Ag+) release from Ag particles was determined by dialysis. The results revealed that AgNPs induce a decrease in the photochemical efficiency of photosystem II (PSII) and an increase in the non-photochemical quenching. The data also revealed that AgNPs affected the stomatal conductance (gs) and CO2 assimilation. Further, AgNPs induced an overproduction of ROS in Vicia faba leaves. Finally, all observed effects were particle diameter-dependent, increasing with the reduction of AgNPs diameter and revealing that AgBulk caused only a small or no changes on plants. In summary, the results point out that AgNPs may negatively affect the photosynthesis process when accumulated in the leaves, and that the NPs themselves were mainly responsible since negligible Ag+ release was detected.