Cellular Internalization and Toxicity of Chitosan Nanoparticles Loaded with Nobiletin in Eukaryotic Cell Models (Saccharomyces cerevisiae and Candida albicans).

This study involved the synthesis and characterization of chitosan nanoparticles loaded with nobiletin (CNpN) and assessed their toxicity and cellular internalization in eukaryotic cell models (Saccharomyces cerevisiae and Candida albicans). Nanoparticles were prepared via the nanoprecipitation method and physicochemically characterized to determine their hydrodynamic diameter using dynamic light scattering (DLS), their surface charge through ζ-potential measurements, and their chemical structure via Fourier-transform infrared spectroscopy (FTIR). The hydrodynamic diameter and ζ-potential of chitosan nanoparticles (CNp) and CNpN were found to be 288.74 ± 2.37 nm and 596.60 ± 35.49 nm, and 34.51 ± 0.66 mV and 37.73 ± 0.19 mV, respectively. The scanning electron microscopy (SEM) images displayed a particle size of approximately 346 ± 69 nm, with notable sphericity for CNpN. FTIR analysis provided evidence of potential imine bonding between chitosan and nobiletin. Membrane integrity damage could be observed in both S. cerevisiae and C. albicans yeast stained with propidium iodide, demonstrating membrane integrity damage caused by CNp and CNpN, where higher concentration treatments inhibited the development of yeast cells. These findings suggest a selective therapeutic potential of CNpN, which could be promising for the development of antifungal and anticancer therapies. This study contributes to understanding the interaction between nanoparticles and eukaryotic cells, offering insights for future biomedical applications.

Synthesis and toxicological study of chitosan-pirul (Schinus molle L.) essential oil nanoparticles on Aspergillus flavus.

In recent years, the study of essential oils as antifungal alternatives and their encapsulation to increase their properties for greater effects has been tested. In this work, nanoparticles of chitosan-Schinus molle L. essential oil (CS-PEO-Np) with a size of 260 ± 31.1 nm were obtained by ionic gelation and evaluated in some growth phases of Aspergillus flavus, a toxigenic fungus. At a concentration of 250 µg/mL of CS-PEO-Np, the A. flavus mycelial growth was inhibited at 97.1% with respect to control, at 96 h of incubation; the germination and viability of spores were inhibited at 74.8 and 40%, respectively, after exposure to 500 µg/mL of these nanomaterials, at 12 h of incubation. The fluorescence images of stained spores with DAPI showed the affectations caused by nanoparticles in the cell membrane, vacuoles and vacuolar content, cell wall, and nucleic acids. For both nanoparticles, CS-Np and CS-PEO-Np, no mutagenic effect was observed in Salmonella Typhimurium; also, the phytotoxic assay showed low-to-moderate toxicity toward seeds, which was dependent on the nanoparticle's concentration. The acute toxicity of CS-PEO-Np to A. salina nauplii was considered low in comparison to CS-Np (control), which indicates that the incorporation of Schinus molle essential oil into nanoparticles of chitosan is a strategy to reduce the toxicity commonly associated with nanostructured materials. The nanoparticulated systems of CS-PEO-Np represent an effective and non-toxic alternative for the control of toxigenic fungi such as A. flavus by delaying the initial growth stage.

Effect of a Prolonged-Release System of Urea on Nitrogen Losses and Microbial Population Changes in Two Types of Agricultural Soil.

Urea is the nitrogen-containing fertilizer most used in agricultural fields; however, the nutrient given by the urea is lost into the environment. The aim of this research was to determine the effect of two soil textures by applying a prolonged-release system of urea (PRSU) on the N losses. This research shows an important decrease of the nitrate and ammonium losses from 24.91 to 87.94%. Also, the microbiological population increases after the application of the PRSU. It was concluded that both soil textures presented the same loss-reduction pattern, where the N from the nitrates and ammonium was reduced in the leachates, increasing the quality of the soil and the microbial population in both soil textures after the PRSU application.

Nanoparticles of Betalain-Gelatin with Antioxidant Properties by Coaxial Electrospraying: Preparation and Characterization.

Betalains are bioactive compounds with attractive antioxidant properties for the food industry, endowing them with potential application in food coatings to maintain quality and extend shelf life. However, they have low stability to factors such as light, temperature, and humidity. An alternative to protect bioactive compounds is nanoencapsulation; one of the most used techniques to produce an encapsulation is coaxial electrospraying. In this research, the preparation and characterization of gelatin-betalain nanoparticles were carried out using the coaxial electrospray technique. Betalains were extracted from pitaya (Stenocereus thurberi) and encapsulated in gelatin. The obtained material was evaluated by SEM, FTIR, TGA, and DSC techniques and for its antioxidant capacity. By SEM, nanoparticles with spherical and monodisperse morphologies were observed, with betalain concentrations of 1 and 3% w/v and average diameters of 864 and 832 µm, respectively. By FTIR, the interaction between betalain and gelatin was observed through amino groups and hydrogen bonds. Likewise, the antioxidant activity of the betalains was maintained at the time of encapsulation, increasing the antioxidant activity as the concentration increased. The results of the DPPH, ABTS, and total phenols methods were 645.4592 µM T/g, 832.8863 ± 0.0110 µM T/g, and 59.8642 ± 0.0279 mg GAE/g for coaxial nanoparticles with 3% betalains, respectively. Therefore, the coaxial electrospray technique was useful for obtaining nanoparticles with good antioxidant properties, and due to the origin of its components and since the use of toxic solvents is not necessary in the technique, the material obtained can be considered food grade with potential application as a coating on functional foods.

Tetrandrine Derivatives as Promising Antibacterial Agents.

This work reports on the antibacterial activity of two tetrandrine derivatives, with acridine (MAcT) and anthracene (MAnT) units, against Gram-positive and Gram-negative bacteria of clinical importance by the broth microdilution method as well as their antioxidant activity against ABTS•+ and DPPH•+ radicals. Unlike natural tetrandrine, its derivatives inhibited bacterial growth, showing selectivity against Staphylococcus aureus with notable activity of MAnT (MIC = 0.035 µg/mL); this compound also has good activity against the ABTS•+ radical (IC50 = 4.59 µg/mL). Cell membrane integrity studies and reactive oxygen species (ROS) detection by fluorescent stains helped to understand possible mechanisms related to antibacterial activity, while electrophoretic mobility assays showed that the derivatives can bind to bacterial DNA plasmid. The results indicate that MAnT can induce a general state of oxidative stress in S. aureus and Escherichia coli, while MAcT induces an oxidative response in S. aureus. Complementary electrochemical studies were included.

Properties of Cephalopod Skin Ommochromes to Inhibit Free Radicals, and the Maillard Reaction and Retino-Protective Mechanisms in Cellular Models Concerning Oxidative Stress, Angiogenesis, and Inflammation.

Ommochromes are pigments of invertebrates that exhibit oxidative stress protection. The aim of this study was to investigate ommochromes extracted from cephalopod's skin for their ability to inhibit age-related-macular degeneration (AMD)-related factors such as H2O2-induced and iron-dependent oxidative stress (ferroptosis and erastin), accumulation of advanced glycation end-products (AGEs), as well as vascular endothelial growth factor (VEGF), and inflammatory cytokines (interleukin 6 and interleukin 8) secretion. As cell systems, we used primary porcine retinal pigment epithelium (RPE), human retinal pigment epithelium cell line ARPE-19 and uveal melanoma cell line OMM-1. In vitro, ommochromes produced an antiglycation effect by the inhibition of fructosylation reaction. The ommochromes showed protective effects against erastin- induced cell death in ARPE-19. In addition, in long-term stimulation (7 days) ommochromes decreased constitutively secreted VEGF, as well as interleukin 6 and interleukin 8 induced by Poly I:C in primary RPE. No relevant effects were detected in OMM-1 cells. The effects are dependent on the cell system, time of exposition, and concentration. This substance is of interest for further research concerning age-related macular degeneration.

Inhibition of Pathogenic Bacteria and Fungi by Natural Phenoxazinone from Octopus Ommochrome Pigments.

Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

Relationship between the Antifungal Activity of Chitosan-Capsaicin Nanoparticles and the Oxidative Stress Response on Aspergillus parasiticus.

The fungus Aspergillus parasiticus is a contaminant in agricultural crops and its eradication involves the indiscriminate use of harmful synthetic pesticides. In the search for antifungal agents of natural origin, chitosan (Q) and capsaicin (C) are coupled in the form of nanoparticles (Np), which can possess a direct application under specific conditions. Due to their small size, Np can cross through the cell wall, taking the cells into a pro-oxidant environment known as "oxidative stress", which presents when the reactive oxygen species (ROS) surpass the number of antioxidants in the cell. In the present investigation, nanoparticles of chitosan (Np Q) and nanoparticles of chitosan-capsaicin (Np QC) with an average diameter of 44.8 ± 20.6 nm and 111.1 ± 14.1 nm, respectively, were synthesized, and there was a zeta potential of + 25.6 ± 0.7 mV and + 26.8 ± 6.1 mV, respectively. The effect of the concentration of Np Q (A, B, C, and D), of Np QC (A, B, C, and D), and capsaicin in a solution (control) was evaluated on the viability of the spores, the accumulation of intracellular ROS, and the morphometric changes of A. parasiticus. Acute toxicity of the Np was determined utilizing bioassays with Artemia salina, and acute phytotoxicity was evaluated in lettuce seeds (Lactuca sativa). According to ROS results, capsaicin (control) did not induce oxidative stress in the cell; otherwise, it was observed to have an elevated (p < 0.05) accumulation of ROS when the concentration of Np Q increased. For both, Np Q and Np QC, an inverse physiological pattern relating spore viability and ROS accumulation in the fungus was found; the viability of spores decreased as the ROS accumulation increased. The spore viability of A. parasiticus diminished upon increasing the concentration of chitosan (0.3−0.4 mg/mL) in the Np, while the intracellular accumulation of ROS increased proportionally to the concentration of the nanomaterials in the treatments of Np Q and Np QC. On the other hand, Np QC presented a lower (p < 0.05) toxicological effect in comparison with Np Q, which indicates that the incorporation of bioactive compounds, such as capsaicin, into nanoparticles of chitosan is a strategy that permits the reduction of the toxicity associated with nanostructured materials.

Nutraceuticals/Drugs Promoting Mitophagy and Mitochondrial Biogenesis May Combat the Mitochondrial Dysfunction Driving Progression of Dry Age-Related Macular Degeneration.

In patients with age-related macular degeneration (AMD), the crucial retinal pigment epithelial (RPE) cells are characterized by mitochondria that are structurally and functionally defective. Moreover, deficient expression of the mRNA-editing enzyme Dicer is noted specifically in these cells. This Dicer deficit up-regulates expression of Alu RNA, which in turn damages mitochondria-inducing the loss of membrane potential, boosting oxidant generation, and causing mitochondrial DNA to translocate to the cytoplasmic region. The cytoplasmic mtDNA, in conjunction with induced oxidative stress, triggers a non-canonical pathway of NLRP3 inflammasome activation, leading to the production of interleukin-18 that acts in an autocrine manner to induce apoptotic death of RPE cells, thereby driving progression of dry AMD. It is proposed that measures which jointly up-regulate mitophagy and mitochondrial biogenesis (MB), by replacing damaged mitochondria with "healthy" new ones, may lessen the adverse impact of Alu RNA on RPE cells, enabling the prevention or control of dry AMD. An analysis of the molecular biology underlying mitophagy/MB and inflammasome activation suggests that nutraceuticals or drugs that can activate Sirt1, AMPK, Nrf2, and PPARα may be useful in this regard. These include ferulic acid, melatonin urolithin A and glucosamine (Sirt1), metformin and berberine (AMPK), lipoic acid and broccoli sprout extract (Nrf2), and fibrate drugs and astaxanthin (PPARα). Hence, nutraceutical regimens providing physiologically meaningful doses of several or all of the: ferulic acid, melatonin, glucosamine, berberine, lipoic acid, and astaxanthin, may have potential for control of dry AMD.

HALOBACTERIAS PROMOTORAS DE CRECIMIENTO VEGETAL ASOCIADAS A Lippia palmer! (VERBENACEAE) EN LA ZONA ÁRIDA DEL NOROESTE DE MÉXICO / Plant growth promoting halobacteria associated to plant growth promoting halobacteria associated to Lippia palmeri (Verbenaceae) in the arid zone of northwestern Mexico

RESUMEN La producción de orégano es de relevancia económica en el noroeste de México. Sin embargo, los productores para obtener altos rendimientos recurren a la fertilización química, pero su mal uso, agudiza la salinidad. Lippia palmeri crece de manera natural en suelos áridos, pobres en materia orgánica, alta salinidad y temperatura en el noroeste de México. En el contexto de una agricultura sustentable, los microorganismos mantienen la fertilidad del suelo e incrementan la productividad de la planta. Actualmente existe interés en proponer biofertilizantes en la agricultura de alta intrusión salina y elevadas temperaturas para el cultivo de orégano. Las Halobacterias Promotoras del Crecimiento de Plantas (HPCP), se han destacado por beneficiar a los cultivos nutrimentalmente y mitigar el efecto de la salinidad. El objetivo del presente trabajo consistió en identificar termo- y halo-tolerantes HPCP asociadas a la rizosfera de L. palmeri; se evaluó la actividad solubilizadora de fosfatos, producción de ácidos orgánicos, sideróforos y fijación de nitrógeno; se identificaron mediante el gen ARNr-16S aquellas con alta actividad evaluándose su efecto en la germinación y longitud radicular. Quince diferentes colonias sobresalieron al crecer en NaCl (0.25, 0.50 y 0.75 M) a 35 y 45 °C, destacando tres bacterias identificadas: Bacillus amyloliquefaciens, Bacillus subtilis y Bacillus licheniformis. El efecto en la longitud radicular es significativo por la aplicación de B. amyloliquefaciens. Estudios relacionados con la promoción vegetal deben ser considerados en posteriores estudios. Este es el primer informe de B. amyloliquefaciens como una bacteria fijadora de nitrógeno asociada a L. palmeri.

ABSTRACT The production of oregano is of economic relevance in northwestern Mexico. However, to obtain high yields, producers resort to chemical fertilization, but its misuse increases salinity. Lippia palmeri Watts is a species of oregano that naturally grows in arid soils with poor organic matter, high salinity and temperature in the northwestern Mexico. In the context of sustainable agriculture, microorganisms activate soil fertility and increase plant productivity. Currently there is interest in proposing biofertilizers in the agriculture with high saline intrusion and temperatures for the cultivation of oregano. The Plant Growth Promoting Halobacteria (HPCP) have stood out by the beneficiary of the nutritious crops and mitigate the effect of the salinity. The goal of this work was to identify HPCP associated to the rhizosphere of L. palmeri, thermo and halotolerant; phosphate solubilizing activity, organic acid production, siderophores and nitrogen fixation were evaluated; the highest activity colonies were identified by the rRNA-16S gene and the effect on germination and root length was evaluated. Fifteen different colonies stood out when growing in NaCl (G.25, G.5G and G.75 M) at 35 and 45 °C, from which three bacteria were identified: Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis. The effect on root length was significant for B. amyloliquefaciens. Studies related to plant promotion should be involved in subsequent studies. This is the first report of Bacillus amyloliquefaciens as a nitrogen fixing bacteria associated with Lipia palmeri.

Au@Ag Core@Shell Nanoparticles Synthesized with Rumex hymenosepalus as Antimicrobial Agent.

In this work, we used a sequential method of synthesis for gold-silver bimetallic nanoparticles with core@shell structure (Au@AgNPs). Rumex hymenosepalus root extract (Rh), which presents high content in catechins and stilbenes, was used as reductor agent in nanoparticles synthesis. Size distribution obtained by Transmission Electron Microscopy (TEM) gives a mean diameter of 36 ± 11 nm for Au@AgNPs, 24 ± 4 nm for gold nanoparticles (AuNPs), and 13 ± 3 nm for silver nanoparticles (AgNPs). The geometrical shapes of NPs were principally quasi-spherical. The thickness of the silver shell over AuNPs is around 6 nm and covered by active biomolecules onto the surface. Nanoparticles characterization included high angle annular dark field images (HAADF) recorded with a scanning transmission electron microscope (STEM), Energy-Dispersive X-ray Spectroscopy (EDS), X-Ray Diffraction (XRD), UV-Vis Spectroscopy, Zeta Potential, and Dynamic Light Scattering (DLS). Fourier Transform Infrared Spectrometer (FTIR), and X-ray Photoelectron Spectroscopy (XPS) show that nanoparticles are stabilized by extract molecules. A growth kinetics study was performed using the Gompertz model for microorganisms exposed to nanomaterials. The results indicate that AgNPs and Au@AgNPs affect the lag phase and growth rate of Escherichia coli and Candida albicans in a dose-dependent manner, with a better response for Au@AgNPs.

Synthesis and Characterization of a Fe3O4@PNIPAM-Chitosan Nanocomposite and Its Potential Application in Vincristine Delivery.

In this research, we conducted a systematic evaluation of the synthesis parameters of a multi-responsive core-shell nanocomposite (Fe3O4 nanoparticles coated by poly(N-isopropylacrylamide) (PNIPAM) in the presence of chitosan (CS) (Fe3O4@PNIPAM-CS). Scanning electron microscopy (SEM) was used to follow the size and morphology of the nanocomposite. The functionalization and the coating of Fe3O4 nanoparticles (Nps) were evaluated by the ζ-potential evolution and Fourier Transform infrared spectroscopy (FTIR). The nanocomposite exhibited a collapsed structure when the temperature was driven above the lower critical solution temperature (LCST), determined by dynamic light scattering (DLS). The LCST was successfully shifted from 33 to 39 °C, which opens the possibility of using it in physiological systems. A magnetometry test was performed to confirm the superparamagnetic behavior at room temperature. The obtained systems allow the possibility to control specific properties, such as particle size and morphology. Finally, we performed vincristine sulfate loading and release tests. Mathematical analysis reveals a two-stage structural-relaxation release model beyond the LCST. In contrast, a temperature of 25 °C promotes the diffusional release model. As a result, a more in-depth comprehension of the release kinetics was achieved. The synthesis and study of a magnetic core-shell nanoplatform offer a smart material as an alternative targeted release therapy due to its thermomagnetic properties.

Characterization of Epigallocatechin-Gallate-Grafted Chitosan Nanoparticles and Evaluation of Their Antibacterial and Antioxidant Potential.

Nanoparticles based on chitosan modified with epigallocatechin gallate (EGCG) were synthetized by nanoprecipitation (EGCG-g-chitosan-P). Chitosan was modified by free-radical-induced grafting, which was verified by Fourier transform infrared (FTIR). Furthermore, the morphology, particle size, polydispersity index, and zeta potential of the nanoparticles were investigated. The grafting degree of EGCG, reactive oxygen species (ROS) production, antibacterial and antioxidant activities of EGCG-g-chitosan-P were evaluated and compared with those of pure EGCG and chitosan nanoparticles (Chitosan-P). FTIR results confirmed the modification of the chitosan with EGCG. The EGCG-g-chitosan-P showed spherical shapes and smoother surfaces than those of Chitosan-P. EGCG content of the grafted chitosan nanoparticles was 330 µg/g. Minimal inhibitory concentration (MIC) of EGCG-g-chitosan-P (15.6 µg/mL) was lower than Chitosan-P (31.2 µg/mL) and EGCG (500 µg/mL) against Pseudomonas fluorescens (p < 0.05). Additionally, EGCG-g-chitosan-P and Chitosan-P presented higher Staphylococcus aureus growth inhibition (100%) than EGCG at the lowest concentration tested. The nanoparticles produced an increase of ROS (p < 0.05) in both bacterial species assayed. Furthermore, EGCG-g-chitosan-P exhibited higher antioxidant activity than that of Chitosan-P (p < 0.05) in 2,2'-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and ferric-reducing antioxidant power assays. Based on the above results, EGCG-g-chitosan-P shows the potential for food packaging and biomedical applications.

Phytotoxicity, cytotoxicity, and in vivo antifungal efficacy of chitosan nanobiocomposites on prokaryotic and eukaryotic cells.

Chitosan (CS) nanosystems have potential applications for the control of microorganisms in the medical, environmental, and agrifood fields. In vivo and in vitro assays of CS nanosystems have experienced increased activity due to improved physicochemical properties, biological activity, and reactivity. Hence, it is important to determine whether their application involves toxicological risks. The aim of this study was to evaluate the mutagenic, cytotoxic, phytotoxic, and in vivo antifungal activity of chitosan-pyrrole-2-carboxylic acid nanobiocomposites (CS-PCA). The CS-PCA nanoparticles were synthesized by means of the nanoprecipitation technique with a size and ζ-potential of 502 ± 72 nm and + 54.7 ± 15.0 mV, respectively. According to the Ames test, no evidence of mutagenic activity was observed in Salmonella typhimurium strains. The cytotoxic assay showed that the incorporation of PCA into the CS matrix increased the toxic effect on ARPE-19 cells. However, fluorescence microscopy of ARPE-19 cells did not reveal morphostructural changes allusive to cell injury. CS-PCA exhibited strong phytotoxicity on lettuce seeds and the complete inhibition of seed development. The antifungal assay demonstrated that the CS-PCA delayed Aspergillus niger infection in tomato fruit until day 3; however, its use for the pre-treatment of seeds might exert adverse effects on plant development.

Persistence of the antifungal capacity of a fraction of Jacquinia macrocarpa plant against Fusarium verticillioides after continuous exposure.

This study aimed to determine the ability of Fusarium verticillioides in developing mechanisms to counteract the antifungal effect of a fraction from Jacquinia macrocarpa plant extract (JmAF), as well as the morphological and physiological changes that occur during its exposure. The fungus was exposed to JmAF during consecutive periods. A culture sample was taken weekly to determine radial growth, spore germination and size, and fungal ß-1,3-glucanase activity. The results showed that, in the beginning, the radial growth decreased by 85.8%, and spore germination was delayed. As the exposure continued, the fungus showed a recovery, to some extent, in its original characteristics. However, the radial growth of the fungus continued to be inhibited (42.9%) throughout the experiment (7 weeks). The ß-1,3-glucanase activity also was inhibited by 36.4% during the first week of exposure to JmAF. However, the activity was recovered after 7 weeks of exposure.

Proteomic analysis of the inhibitory effect of the butanolic fraction of Jacquinia macrocarpa on Fusarium verticillioides.

Jacquinia macrocarpa, a plant native to northwestern Mexico, has an inhibitory effect against phytopathogenic fungi. Previous studies have shown that the butanolic extract of J. macrocarpa causes retardation and atrophy in mycelial growth of Fusarium verticillioides. However, the action mechanism of this extract is unknown. We used a proteomics approach to understand the inhibitory effect of J. macrocarpa butanolic extract, based on differential protein accumulation in F. verticillioides. Proteins were extracted from F. verticillioides cultured in Czapek broth with and without 202.12 µg/mL (IC50) of butanolic extract of J. macrocarpa. Thirty-eight protein spots showing statistically significant changes (ANOVA, p < 0.01) and at least a 2-fold change in abundance between experimental conditions were analyzed by mass spectrometry. Identified proteins were grouped into different biological processes according to Gene Ontology, among them were amino acid metabolism, protein folding and stabilization, protein degradation, protein transport, carbohydrate metabolism, oxidative stress response, and miscellaneous. This work is the first report of changes in the proteomic profile of F. verticillioides exposed to the J. macrocarpa extract. This information provides new insights into the inhibitory mechanism of the extract and represents a starting point for dissection of the fungal response against the J. macrocarpa extract components.

Biosorption of copper by immobilized biomass of Aspergillus australensis. Effect of metal on the viability, cellular components, polyhydroxyalkanoates production, and oxidative stress.

Heavy metals are toxic especially when they are introduced into the environment due to anthropogenic activities such as metallurgy, mining, and tanning. Removing these pollutants has become a worldwide concern since they cannot be degraded into nontoxic forms causing extended effects in the ecosystems. The use of an Aspergillus australensis was evaluated in order to remove Cu2+ from simulated wastewater. The fungus was isolated from river sludges contaminated with heavy metals and was first evaluated for the determination of Cu2+ tolerance levels. Microscopic fluorescence analysis was carried out to determine the effect of Cu2+ presence on the viability, cellular components, polyhydroxyalkanoates production, and oxidative stress of the fungus, as a response to the stress caused by exposure to metal. In order to achieve copper removal, the A. australensis biomass was produced using batch cultures, and the mycelium was immobilized on a textile media in order to compare the copper-removal efficiency of live or dead biomass. The optimal values of pH and temperature for biomass production were established by using a surface response analysis. Live immobilized biomass was capable of removing Cu2+ from 1.54 ± 0.19 to 2.66 ± 0.26 mg of copper/ g of dry biomass, while values of 1.93 ± 0.03 to 2.36 ± 0.29 mg of copper/g of dry biomass were observed when dead biomass was used. As was expected, copper removal using biomass varied depending on the pH and temperature used.

Inhibition of the antioxidant activity of catalase and superoxide dismutase from Fusarium verticillioides exposed to a Jacquinia macrocarpa antifungal fraction.

The aim of this study was to investigate the in vitro effect of an antifungal fraction obtained from Jacquinia macrocarpa plant (JmAF) in the generation of reactive oxygen species (ROS) and the activity of the catalase (CAT) and superoxide dismutase (SOD) enzymes from Fusarium verticillioides, as well as their influence in the viability of the fungus spores. The compounds present in the JmAF were determined by gas chromatography/quadrupole time-of-flight mass spectrometry (GC/QTOF-MS). The effect of the exposition to JmAF on the generation of ROS, as well as in the CAT and SOD activities in F. verticillioides, was determined. The main compounds detected were γ-sitosterol, stephamiersine, betulinol and oleic acid. JmAF showed very high ability in inhibiting the spore viability of F. verticillioides, and their capacity to cause oxidative stress by induction of ROS production. JmAF induced the highest ROS concentration and also inhibited CAT and SOD activities. The results obtained in this study indicate that JmAF is worthy of being considered for the fight against phytopathogenic fungi.

Toxicological Assessment of Cross-Linked Beads of Chitosan-Alginate and Aspergillus australensis Biomass, with Efficiency as Biosorbent for Copper Removal.

Sorbent materials of biological origin are considered as an alternative to the use of traditional methods in order to remove heavy metals. Interest in using these materials has increased over the past years due to their low cost and friendliness to the environment. The objective of this study was to synthesize and characterize cross-linked beads made of chitosan, alginate, and mycelium of a copper-tolerant strain of Aspergillus australensis. The acute toxicity of the biocomposite beads was assessed using brine shrimp Artemia salina nauplii and the phytotoxicity was determined using lettuce (Lactuca sativa) and chili pepper 'Anaheim' (Capsicum annuum) seeds. The biosorption capacity for copper removal in simulated wastewater was also evaluated. Results showed that the biosorbent obtained had a maximal adsorption of 26.1 mg of Cu2+ per g of biocomposite, and removal efficiency was around 79%. The toxicity of simulated residual water after treatment with the biocomposite showed low toxicity toward seeds, which was highly dependent on the residual copper concentration. The toxicity of the biocomposite beads to A. salina was considered medium depending on the amount of the biocomposite, which was attributed to low pH. Biocomposite shows promise as biosorbent for the removal process of heavy metals.

Synthesis of chitosan biocomposites loaded with pyrrole-2-carboxylic acid and assessment of their antifungal activity against Aspergillus niger.

A wide variety of chitosan (CS) biomaterials have been loaded with different antimicrobial agents to improve the activity of CS against phytopathogenic fungi. Recently, the antimicrobial activity of 1H-pyrrole-2-carboxylic acid (PCA) has been reported as a secondary metabolite of Streptomyces griseus, which was identified as the main bioactive compound in the biological control. However, it is sensitive to light and its activity against filamentous fungi has not yet been reported. The aim of the present research work was to evaluate the biological activity of CS-PCA biocomposites for the control of Aspergillus niger. CS-PCA biocomposites were obtained through nanoprecipitation. In vitro antifungal activity was determined by viability assay, spore germination, morphometric analysis of spores and hyphae, and the analysis of cellular components by fluorescence microscopy. CS-PCA showed an average size and Z potential of 502 ± 72 nm and + 54.7 ± 15 mV, respectively. Micrographs demonstrated well-distributed biocomposites with an apparently spherical shape. A new signal at 1473 cm-1 in the FT-IR spectrum of the CS-PCA biocomposite was observed, confirming the presence of PCA in the composition of the CS-PCA nanosystem. CS-PCA biocomposites reduced the spores' viability by up to 58%. Effects on fungi morphometry, observed as an increase in the spores' average diameter, swelling, distortion, and an increase in the branching of hyphae, were observed. Fluorescence analysis showed oxidative stress and membrane and cell wall damage, mainly at early growth stages. The inhibitory effect against CS-resistant fungi, such as A. niger, opens a door for the control of CS-sensitive fungi.