Polianthes tuberosa-Mediated Silver Nanoparticles from Flower Extract and Assessment of Their Antibacterial and Anticancer Potential: An In Vitro Approach.

Plant-mediated metallic nanoparticles have beenreported for a diversified range of applications in biological sciences. In the present study, we propose the Polianthes tuberosa flower as a reducing and stabilizing agent for the synthesis of silver nanoparticles (PTAgNPs). The PTAgNPs were exclusively characterized using UV-Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), atomic force microscopy, zeta potential, and transmission electron microscopy (TEM) studies. In a biological assay, we investigated the antibacterial and anticancer activity of silver nanoparticles in the A431 cell line. The PTAgNPs demonstrated a dose-dependent activity in E. coli and S. aureus, suggesting the bactericidal nature of AgNPs. The PTAgNPs exhibited dose-dependent toxicity in the A431 cell line, with an IC50 of 54.56 µg/mL arresting cell growth at the S phase, as revealed by flow cytometry analysis. The COMET assay revealed 39.9% and 18.15 severities of DNA damage and tail length in the treated cell line, respectively. Fluorescence staining studies indicate that PTAgNPs cause reactive oxygen species (ROS) and trigger apoptosis. This research demonstrates that synthesized silver nanoparticles have a significant effect on inhibiting the growth of melanoma cells and other forms of skin cancer. The results show that these particles can cause apoptosis or cell death in malignant tumor cells. This suggests that they could be used to treat skin cancers without harming normal tissues.

Rhizobium pusense-Mediated Selenium Nanoparticles-Antibiotics Combinations against Acanthamoeba sp.

Severe ocular infections by Acanthamoeba sp. lead to keratitis, resulting in irreversible vision loss in immune-compromised individuals. When a protozoal infection spreads to neural tissues, it causes granulomatous encephalitis, which can be fatal. Treatment often takes longer due to the transition of amoeba from trophozoites to cyst stages, cyst being the dormant form of Acanthamoeba. A prolonged use of therapeutic agents, such as ciprofloxacin (Cipro), results in severe side effects; thus, it is critical to improve the therapeutic efficacy of these widely used antibiotics, possibly by limiting the drug-sensitive protozoal-phase transition to cyst formation. Owing to the biomedical potential of selenium nanoparticles (SeNPs), we evaluated the synergistic effects of ciprofloxacin and Rhizobium pusense-biogenic SeNPs combination. SeNPs synthesized using Rhizobium pusense isolated from root nodules were characterized using UV-Visible spectrophotometer, FT-IR, SEM with EDX, particle size analysis, and Zeta potential. The combination was observed to reduce the sub-lethal dose of Cipro, which may help reduce its side effects. The selenium and ciprofloxacin (SeNPs-Cipro) combination reduced the LC50 by 33.43%. The anti-protozoal efficacy of SeNPs-Cipro was found to transduce through decreased protozoal-cyst formations and the inhibition of the galactosidase and protease enzymes of trophozoites. Furthermore, high leakage of sugar, proteins, and amino acids during the SeNPs-Cipro treatment was one primary reason for killing the trophozoites. These experimental results may be helpful in the further pre-clinical evaluation of SeNPs-Cipro to combat protozoal infections. Future studies for combinations of SeNPs with other antibiotics need to be conducted to know the potential of SeNPs against antibiotic resistance in Acanthamoeba.

Biotechnology methods for succession of bacterial communities in polychlorinated biphenyls (PCBs) contaminated soils and isolation novel PCBs-degrading bacteria.

Polychlorinated Biphenyls (PCBs) are persistence in the contaminated sites as a result of lacking PCBs-degrading microorganisms. Cultivation-independent technique called single-strand-conformation polymorphism (SSCP) based on 16SrRNA genes was chosen to characterize the diversity of bacterial communities in PCBs polluted soil samples. The bacterial communities showed an increasing diversity from the genetic profiles using SSCP technique. 51 single products were identified from the profiles using PCR reamplification and cloning. DNA sequencing of the 51 products, it showed similarities to Acidobacteria, Actinobacteria, Betaproteobateria, Gammaproteobacteria and Alphaproteobacteria, the range of similarities were 92.3 to 100%. Pure 23 isolates were identified from PCBs contaminated sites. The identified isolates belonged to genus Bacillus, Brevibacillus, Burkholderia, Pandoraea, Pseudomonas, and Rhodococcus. The new strains have the capability to use PCBs as a source of sole carbon and harbor 2,3-dihydroxybiphenyl dioxygenase (DHBDO) which could be used as molecular marker for detection PCBs-degrading bacteria in the PCBs contaminated sites. This finding may enhance the PCBs bioremediation by monitoring and characterization of the PCBs degraders using DHBDO in PCBs contaminated sites.

Ethnomedicinal Plants with Protective Effects against Beta-Amyloid Peptide (Aß)1-42 Indicate Therapeutic Potential in a New In Vivo Model of Alzheimer's Disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder with unmet medical need. This investigation consisted of testing a range of ethanolic ethnomedicinal plant extracts (n = 18) traditionally used in the treatment of disorders such as anxiety, delirium, and memory loss. They were then screened for in vitro inhibitory activity against acetylcholinesterase (AChE), butylcholinesterase (BuChE), beta-secretase 1/beta-site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1), and antioxidant activities. Plants with potent activities were further characterised using a recently developed in vivo model of AD, Globodera pallida. The ability of phytoextracts to protect this organism against amyloid-beta Aß (1-42) exposure was assessed by measuring chemosensing, survival rate, production of reactive oxygen species (ROS), and antioxidant responses. Extracts (n = 5) from Juglans regia (leaves), Ellettaria cardamomum (seeds), Cinnamomum zeylanicum (bark), Salvia officinalis (leaves/flowers), and Hypericum perforatum (flowers) exerted concentration-dependent inhibitory activities against AChE and BuChE. Three of these plant extracts (i.e., J. regia, E. cardamomum, and S. officinalis) possessed strong concentration-dependent inhibitory activity against BACE1. Furthermore, the five selected medicinal plant extracts not only enhanced significantly (p < 0.05) the nematode's chemosensing, survival rate, and antioxidant responses (i.e., anti-ROS production, mitochondrial reductase activity, oxidized glutathione (GSSG) to reduced glutathione (GSH) ratio), but also greatly restored (p < 0.05) in a concentration-dependent manner the Aß (1-42)-induced deleterious changes in these same parameters. In brief, this investigation highlights plant extracts with strong anti-AD activities which could be trialled as novel therapeutic supplements or undergo further biodiscovery research.

Application of Nanocomposites from Bees Products and Nano-Selenium in Edible Coating for Catfish Fillets Biopreservation.

Bee products, e.g., chitosan and propolis (Pro), have extraordinary importance in many disciplines including food biopreservation. Fish meat is highly susceptible to vast spoilage, especially catfish (Clarias gariepinus) products. The current work involved the extraction of bees' chitosan nanoparticles (BCht), Pro, Pro-mediated SeNPs and their composites, to evaluate them as potential antimicrobial and preservative nano-compounds, for the preservation of catfish fillets and augment their quality. BCht was extracted from bees (Apis mellifera) corpses and had a 151.9 nm mean particle diameter. The Pro was used for biosynthesis of SeNPs, which had 11.2 nm mean diameters. The entire compounds/composites exhibited powerful antibacterial acts against Escherichia coli, Staphylococcus aureus and Salmonella typhimurium, where S aureus had the uppermost resistance. BCht/Pro/SeNPs were the most forceful toward all bacterial strains. The constructed edible coatings (ECs) from produced compounds/composites (BCht, Pro, Pro/SeNPs, Pro/BCht and BCht/Pro/SeNPs) had elevated efficiency for preserving catfish fillets during cold storages for 7 days. The microbiological (total counts, psychrophilic bacteria, yeast and molds), spoilage chemical parameters (TVB-N, TBARS) and sensorial attributes (appearance, odor, color, overall quality) of ECs-treated fillets indicated the nanocomposite's efficiency for protecting the fish from microbial growth, the progress of chemical spoilage indicators and maintaining the sensorial quality of treated stored fillets. The most effective nanocomposite for maintaining the entire fillet's quality was the BCht/Pro/SeNP. The based ECs on BNCt, Pro/SeNPs and their nanocomposites could be endorsed for prospective employment in the biopreservation of various seafoods.

Elicitor-Induced Metabolomics Analysis of Halodule pinifolia Suspension Culture for an Alternative Antifungal Screening Approach against Candida albicans.

Elicitors are the agents that stimulate the defense responses of plants, and accumulate specialized metabolites in plant tissue culture. This study investigated the elicitor-feeding response of H. pinifolia suspension cell cultures (SCC) for metabolomics analysis and screening of specialized compounds against Candida albicans. Methyl jasmonate (MeJA) was used as an elicitor, and treatment of SCC at a concentration of 20 µM MeJA resulted in the maximum rosmarinic acid (RA) accumulation (117 mg/g dry weight), with transcript levels of RA biosynthetic genes HpPAL, HpC4H, and Hp4CL being 4.2, 2.5, and 3.7-fold higher, respectively, than the controls. GC-MS-based metabolomics analysis revealed a total of 47 metabolites, including 30 organic acids, six amino acids, six flavonoids, two sugars, two plant growth regulators, and one vitamin, which were significantly different between control and MeJA-treated cells. Furthermore, five phenolic acids were discovered at higher concentrations, including p-anisic acid, p-coumaric acid, caffeic acid, vanillic acid, and rosmarinic acid, and were purified and structurally elucidated for alternative antifungal screening against C. albicans and the evaluation of ADMET properties. The results from antifungal screening revealed that RA at MIC of 31.25 mg/L exhibited the lowest growth percentage of C. albicans (1.99%), with higher inhibition of isocitrate lyase 1 (ICL 1) enzyme (93.1%), followed by p-anisic acid (86.2%) and caffeic acid (85.1%), respectively. The drug likeliness and ADMET properties of RA exhibited promising results, with a bioactivity score of 0.57, 0.15, and 0.24 for nuclear receptor ligand, protease inhibitor, and enzyme inhibitor, respectively. Therefore, MeJA appears to have a significant effect on enhanced RA accumulation in H. pinifoia cells with phenylpropanoid transcript expression, and acts as an ICL1 inhibitor of C. albicans.

Prediction of Heterosis for Agronomic Traits in Half-Diallel Cross of Rice (Oryza sativa L.) under Drought Stress Using Microsatellite Markers.

Rice breeders are encouraged to classify potential F1-hybrids without crossing all viable mixtures by testing depending on genetic distance due to lack of labor and time in yield trials. The goals of this research were to establish heterosis and investigate the relationship between genomic distance and agronomic attributes under drought. Half-diallel mating design, 28 F1's and 8 parents were evaluated under drought and genotyped using 11 microsatellite markers. In total, 39 alleles were detected. Results indicated that the greatest heterotic effects for grain yield were observed in Sakha 103 × Sakha 104 and GZ7576-10-3-2-1 × Giza 179, which gave 29.32−22.57% heterosis, respectively. Heterosis for grain yield in these crosses occurred as a rise in panicle weight, filled grains per panicle, low sterility and 100-grain weight. Correlations of marker-based genetic distance with mid-parent heterosis were positively and significantly correlated with sterility percentage (r = 0.390 *, p < 0.05). However, better-parent heterosis was positively and significantly correlated with sterility percentage (r = 0.352 *, p < 0.05) and grain yield per plant (r = 0.345 *, p < 0.05). Associations indicate that high grain yield and low sterility of rice crosses can be expected from microsatellite marker-defined distances of parents. This study indicated that genetic distance is very effective for heterosis prediction in breeding programs.

Isolation and Characterization a Novel Catabolic Gene Cluster Involved in Chlorobenzene Degradation in Haloalkaliphilic Alcanivorax sp. HA03.

Chlorobenzene (CB) poses a serious risk to human health and the environment, and because of its low degradation rate by microorganisms, it persists in the environment. Some bacterial strains can use CB as growth substrates and their degradative pathways have evolved; very little is known about these pathways and the enzymes for CB degradation in high pH and salinity environments. Alcanivorax sp. HA03 was isolated from the extremely saline and alkaline site. HA03 has the capability to degrade benzene, toluene and chlorobenzene (CB). CB catabolic genes were isolated from HA03, which have a complete gene cluster comprising α and ß subunits, ferredoxin and ferredoxin reductase (CBA1A2A3A4), as well as one gene-encoding enzyme for chlorocatechol 1,2-dioxygenase (CC12DOs). Based on the deduced amino acid sequence homology, the gene cluster was thought to be responsible for the upper and lower catabolic pathways of CB degradation. The CBA1A2A3A4 genes probably encoding a chlorobenzene dioxygenase was confirmed by expression during the growth on CB by RT-PCR. Heterologous expression revealed that CBA1A2A3A4 exhibited activity for CB transformation into 3-chlorocatechol, while CC12DOs catalyze 3-chlorocatechol, transforming it into 2-chloromucounate. SDS-PAGE analysis indicated that the sizes of CbA1 and (CC12DOs) gene products were 51.8, 27.5 kDa, respectively. Thus, Alcanivorax sp. HA03 constitutes the first bacterial strain described in the metabolic pathway of CB degradation under high pH and salinity conditions. This finding may have obvious potential for the bioremediation of CB in both highly saline and alkaline contaminated sites.

Growth Regulators Improve Outcrossing Rate of Diverse Rice Cytoplasmic Male Sterile Lines through Affecting Floral Traits.

Cytoplasmic male sterility (CMS) provides an irreplaceable strategy for commercial exploitation of heterosis and producing high-yielding hybrid rice. The exogenous application of plant growth regulators could improve outcrossing rates of the CMS lines by affecting floral traits and accordingly increase hybrid rice seed production. The present study aimed at exploring the impact of growth regulators such as gibberellic acid (GA3), indole-3-acetic acid (IAA), and naphthalene acetic acid (NAA) on promoting floral traits and outcrossing rates in diverse rice CMS lines and improving hybrid rice seed production. The impact of foliar applications of growth regulators comprising GA3 at 300 g/ha or GA3 at 150 g/ha + IAA at 50 g/ha + NAA at 200 g/ha versus untreated control was investigated on floral, growth, and yield traits of five diverse CMS lines. The exogenously sprayed growth regulators, in particular, the combination of GA3, IAA, and NAA (T3) boosted all studied floral, growth, and yield traits in all tested CMS lines. Moreover, the evaluated CMS lines exhibited significant differences in all measured floral traits. L2, L3, and L1 displayed the uppermost spikelet opening angle, duration of spikelet opening, total stigma length, style length, stigma brush, and stigma width. In addition, these CMS lines exhibited the highest plant growth and yield traits, particularly under T3. Consequently, exogenous application of GA3, IAA, and NAA could be exploited to improve the floral, growth, and yield traits of promising CMS lines such as L2, L3, and L1, hence increasing outcrossing rates and hybrid rice seed production.

Trichoderma: An Eco-Friendly Source of Nanomaterials for Sustainable Agroecosystems.

Traditional nanoparticle (NP) synthesis methods are expensive and generate hazardous products. It is essential to limit the risk of toxicity in the environment from the chemicals as high temperature and pressure is employed in chemical and physical procedures. One of the green strategies used for sustainable manufacturing is microbial nanoparticle synthesis, which connects microbiology with nanotechnology. Employing biocontrol agents Trichoderma and Hypocrea (Teleomorphs), an ecofriendly and rapid technique of nanoparticle biosynthesis has been reported in several studies which may potentially overcome the constraints of the chemical and physical methods of nanoparticle biosynthesis. The emphasis of this review is on the mycosynthesis of several metal nanoparticles from Trichoderma species for use in agri-food applications. The fungal-cell or cell-extract-derived NPs (mycogenic NPs) can be applied as nanofertilizers, nanofungicides, plant growth stimulators, nano-coatings, and so on. Further, Trichoderma-mediated NPs have also been utilized in environmental remediation approaches such as pollutant removal and the detection of pollutants, including heavy metals contaminants. The plausible benefits and pitfalls associated with the development of useful products and approaches to trichogenic NPs are also discussed.

Green Synthesis of Gold Nanoparticles Using Polianthes tuberosa L. Floral Extract.

The developments of green-based metallic nanoparticles (gold) are gaining tremendous interest, having potential applications in health care and diagnosis. Therefore, in the present study, Polianthes tuberosa flower filtered extract was used as a reducing and stabilizing agent to synthesize gold nanoparticles (PtubAuNPs). The PtubAuNPs were extensively characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, and X-ray diffraction. The antibacterial activity of PtubAuNPs was determined by the agar well diffusion method; the PtubAuNPs performed extreme antagonistic activity against the tested pathogens. Furthermore, the cytotoxicity of the PtubAuNPs was evaluated in MCF 7 cells by MTT assay. The PtubAuNPs induced toxicity in MCF 7 cells with the least concentration of 100 µg/mL in a dose-dependent method by inducing apoptosis. Overall, the study manifested that PtubAuNPs are a potent nanomaterial that can be employed as an antimicrobial and anticancer agent.

Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants.

Rapid developments in the field of plant genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems necessitate more detailed consideration of the delivery of the CRISPR system into plants. Successful and safe editing of plant genomes is partly based on efficient delivery of the CRISPR system. Along with the use of plasmids and viral vectors as cargo material for genome editing, non-viral vectors have also been considered for delivery purposes. These non-viral vectors can be made of a variety of materials, including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles, as well as nanoscale polymeric materials. They have a decreased immune response, an advantage over viral vectors, and offer additional flexibility in their design, allowing them to be functionalized and targeted to specific sites in a biological system with low cytotoxicity. This review is dedicated to describing the delivery methods of CRISPR system into plants with emphasis on the use of non-viral vectors.

Antifungal Nano-Therapy in Veterinary Medicine: Current Status and Future Prospects.

The global recognition for the potential of nanoproducts and processes in human biomedicine has given impetus for the development of novel strategies for rapid, reliable, and proficient diagnosis, prevention, and control of animal diseases. Nanomaterials exhibit significant antifungal and antimycotoxin activities against mycosis and mycotoxicosis disorders in animals, as evidenced through reports published over the recent decade and more. These nanoantifungals can be potentially utilized for the development of a variety of products of pharmaceutical and biomedical significance including the nano-scale vaccines, adjuvants, anticancer and gene therapy systems, farm disinfectants, animal husbandry, and nutritional products. This review will provide details on the therapeutic and preventative aspects of nanoantifungals against diverse fungal and mycotoxin-related diseases in animals. The predominant mechanisms of action of these nanoantifungals and their potential as antifungal and cytotoxicity-causing agents will also be illustrated. Also, the other theragnostic applications of nanoantifungals in veterinary medicine will be identified.

Conifers Phytochemicals: A Valuable Forest with Therapeutic Potential.

Conifers have long been recognized for their therapeutic potential in different disorders. Alkaloids, terpenes and polyphenols are the most abundant naturally occurring phytochemicals in these plants. Here, we provide an overview of the phytochemistry and related commercial products obtained from conifers. The pharmacological actions of different phytochemicals present in conifers against bacterial and fungal infections, cancer, diabetes and cardiovascular diseases are also reviewed. Data obtained from experimental and clinical studies performed to date clearly underline that such compounds exert promising antioxidant effects, being able to inhibit cell damage, cancer growth, inflammation and the onset of neurodegenerative diseases. Therefore, an attempt has been made with the intent to highlight the importance of conifer-derived extracts for pharmacological purposes, with the support of relevant in vitro and in vivo experimental data. In short, this review comprehends the information published to date related to conifers' phytochemicals and illustrates their potential role as drugs.

Differential Antimycotic and Antioxidant Potentials of Chemically Synthesized Zinc-Based Nanoparticles Derived from Different Reducing/Complexing Agents against Pathogenic Fungi of Maize Crop.

The present study aimed for the synthesis, characterization, and comparative evaluation of anti-oxidant and anti-fungal potentials of zinc-based nanoparticles (ZnNPs) by using different reducing or organic complexing-capping agents. The synthesized ZnNPs exhibited quasi-spherical to hexagonal shapes with average particle sizes ranging from 8 to 210 nm. The UV-Vis spectroscopy of the prepared ZnNPs showed variation in the appearance of characteristic absorption peak(s) for the various reducing/complexing agents i.e., 210 (NaOH and NaBH4), 220 (albumin, and thiourea), 260 and 330 (starch), and 351 nm (cellulose) for wavelengths spanning over 190-800 nm. The FT-IR spectroscopy of the synthesized ZnNPs depicted the functional chemical group diversity. On comparing the antioxidant potential of these ZnNPs, NaOH as reducing agent, (NaOH (RA)) derived ZnNPs presented significantly higher DPPH radical scavenging potential compared to other ZnNPs. The anti-mycotic potential of the ZnNPs as performed through an agar well diffusion assay exhibited variability in the extent of inhibition of the fungal mycelia with maximum inhibition at the highest concentration (40 mg L-1). The NaOH (RA)-derived ZnNPs showcased maximum mycelial inhibition compared to other ZnNPs. Further, incubation of the total genomic DNA with the most effective NaOH (RA)-derived ZnNPs led to intercalation or disintegration of the DNA of all the three fungal pathogens of maize with maximum DNA degrading effect on Macrophomina phaseolina genomic DNA. This study thus identified that differences in size and surface functionalization with the protein (albumin)/polysaccharides (starch, cellulose) diminishes the anti-oxidant and anti-mycotic potential of the generated ZnNPs. However, the NaOH emerged as the best reducing agent for the generation of uniform nano-scale ZnNPs which possessed comparably greater anti-oxidant and antimycotic activities against the three test maize pathogenic fungal cultures.

Nettle-Leaf Extract Derived ZnO/CuO Nanoparticle-Biopolymer-Based Antioxidant and Antimicrobial Nanocomposite Packaging Films and Their Impact on Extending the Post-Harvest Shelf Life of Guava Fruit.

Green synthesized metal oxide nanoparticles (NPs) have prominent applications in antimicrobial packaging systems. Here we have attempted for the fabrication of chitosan-based nanocomposite film containing Urtica dioica leaf extract derived copper oxide (CuO) and zinc oxide (ZnO) NPs for shelf-life extension of the packaged guava fruits. Electron microscopy and spectroscopy analysis of the CuO and ZnO NPs exhibited nano-scale size, spherical morphologies, and negative ζ-potential values. The NPs possessed appreciable antioxidant and antimicrobial activity (AMA) in order of CuO NPs > ZnO NPs >nettle extract. Therefore, this work establishes for the first time the successful synthesis of CuO NPs and compares its antimicrobial and antioxidant properties with ZnO NPs. On incorporation in chitosan, the polymer nanocomposite films were developed by solvent casting technique. The developed films were transparent, had low antioxidant but substantial AMA. The NP supplementation improved the film characteristics as evident from the decrease in moisture content, water holding capacity, and solubility of the films. The nanocomposite films improved the quality attributes and shelf life of guava fruits by one week on packaging and storage compared to unpackaged control fruits. Therefore, this study demonstrates the higher antimicrobial potential of the nettle leaf extract derived CuO/ZnO NPs for development of antimicrobial nanocomposite films as a promising packaging solution for enhancing the shelf life of various perishable fruits.

Nanohybrid Antifungals for Control of Plant Diseases: Current Status and Future Perspectives.

The changing climatic conditions have led to the concurrent emergence of virulent microbial pathogens that attack crop plants and exhibit yield and quality deterring impacts on the affected crop. To counteract, the widespread infections of fungal pathogens and post-harvest diseases it is highly warranted to develop sustainable techniques and tools bypassing traditional agriculture practices. Nanotechnology offers a solution to the problems in disease management in a simple lucid way. These technologies are revolutionizing the scientific/industrial sectors. Likewise, in agriculture, the nano-based tools are of great promise particularly for the development of potent formulations ensuring proper delivery of agrochemicals, nutrients, pesticides/insecticides, and even growth regulators for enhanced use efficiency. The development of novel nanocomposites for improved management of fungal diseases can mitigate the emergence of resilient and persistent fungal pathogens and the loss of crop produce due to diseases they cause. Therefore, in this review, we collectively manifest the role of nanocomposites for the management of fungal diseases.

Copper-Chitosan Nanocomposite Hydrogels Against Aflatoxigenic Aspergillus flavus from Dairy Cattle Feed.

The integration of copper nanoparticles as antifungal agents in polymeric matrices to produce copper polymer nanocomposites has shown excellent results in preventing the growth of a wide variety of toxigenic fungi. Copper-chitosan nanocomposite-based chitosan hydrogels (Cu-Chit/NCs hydrogel) were prepared using a metal vapor synthesis (MVS) and the resulting samples were described by transmission electron microscopy (TEM), X-ray fluorescence analysis (XRF), and small-angle X-ray scattering (SAXS). Aflatoxin-producing medium and VICAM aflatoxins tests were applied to evaluate their ability to produce aflatoxins through various strains of Aspergillus flavus associated with peanut meal and cotton seeds. Aflatoxin production capacity in four fungal media outlets revealed that 13 tested isolates were capable of producing both aflatoxin B1 and B2. Only 2 A. flavus isolates (Af11 and Af 20) fluoresced under UV light in the A. flavus and parasiticus Agar (AFPA) medium. PCR was completed using two specific primers targeting aflP and aflA genes involved in the synthetic track of aflatoxin. Nevertheless, the existence of aflP and aflA genes indicated some correlation with the development of aflatoxin. A unique DNA fragment of the expected 236 bp and 412 bp bands for aflP and aflA genes in A. flavus isolates, although non-PCR fragments have been observed in many other Aspergillus species. This study shows the antifungal activity of Cu-Chit/NCs hydrogels against aflatoxigenic strains of A. flavus. Our results reveal that the antifungal activity of nanocomposites in vitro can be effective depending on the type of fungal strain and nanocomposite concentration. SDS-PAGE and native proteins explain the apparent response of cellular proteins in the presence of Cu-Chit/NCs hydrogels. A. flavus treated with a high concentration of Cu-Chit/NCs hydrogels that can decrease or produce certain types of proteins. Cu-Chit/NCs hydrogel decreases the effect of G6DP isozyme while not affecting the activity of peroxidase isozymes in tested isolates. Additionally, microscopic measurements of scanning electron microscopy (SEM) showed damage to the fungal cell membranes. Cu-Chit/NCS hydrogel is an innovative nano-biopesticide produced by MVS is employed in food and feed to induce plant defense against toxigenic fungi.

Silver/Chitosan Nanocomposites: Preparation and Characterization and Their Fungicidal Activity against Dairy Cattle Toxicosis Penicillium expansum.

This work aimed to evaluate the fungicide activity of chitosan-silver nanocomposites (Ag-Chit-NCs) against Penicillium expansum from feed samples. The physicochemical properties of nanocomposites were characterized by X-ray fluorescence analysis (XRF), small-angle X-ray scattering (SAXS), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM). The morphological integrity of the nanohybrid was confirmed by electron transmission. By the data of RFA (X-ray fluorescent analysis), the contents of Ag in Ag-chitosan composite were 5.9 w/w%. The size distribution of the Ag nanoparticles incorporated in the chitosan matrix was investigated by SAXS. The main part of the size heterogeneity distribution in the chitosan matrix corresponds to the portion of small particles (3-4 nm). TEM analysis revealed a spherical morphology in the form of non-agglomerated caps, and 72% of the nanoparticles measured up to 4 nm. The minimum inhibitory concentration of NCs was evaluated in petri dishes. Three different concentrations were tested for antifungal activity against the mycotoxigenic P. expansum strain. Changes in the mycelium structure of P. expansum fungi by scanning electron microscopy (SEM) were observed to obtain information about the mode of action of Ag-Chit-NCs. It was shown that NC-Chit-NCs with sizes in the range from 4 to 10 nm have internalized sizes in cells, form agglomerates in the cytoplasm, and bind to cell organelles. Besides, their ability to influence protein and DNA fragmentation was examined in P. expansum. SDS-PAGE explains the apparent cellular protein response to the presence of various Ag-Chit-NCs. The intensity of P. expansum hyphal cell protein lines treated with Ag-Chit-NCs was very thin, indicating that high molecular weight proteins are largely prevented from entering the electrophoretic gel, which reflects cellular protein modification and possible damage caused by the binding of several protein fragments to Ag-Chit-NCs. The current results indicate that Ag-Chit-NCs <10 nm in size have significant antifungal activity against P. expansum, the causative agent of blue mold-contaminated dairy cattle feed.