Optimization of pullulan production by Aureobasidium pullulans using semi-solid-state fermentation and artificial neural networks: Characterization and antibacterial activity of pullulan impregnated with Ag-TiO2 nanocomposite.

This study presents a novel and efficient approach for pullulan production using artificial neural networks (ANNs) to optimize semi-solid-state fermentation (S-SSF) on faba bean biomass (FBB). This method achieved a record-breaking pullulan yield of 36.81 mg/g within 10.82 days, significantly exceeding previous results. Furthermore, the study goes beyond yield optimization by characterizing the purified pullulan, revealing its unique properties including thermal stability, amorphous structure, and antioxidant activity. Energy-dispersive X-ray spectroscopy and scanning electron microscopy confirmed its chemical composition and distinct morphology. This research introduces a groundbreaking combination of ANNs and comprehensive characterization, paving the way for sustainable and cost-effective pullulan production on FBB under S-SSF conditions. Additionally, the study demonstrates the successful integration of pullulan with Ag@TiO2 nanoparticles during synthesis using Fusarium oxysporum. This novel approach significantly enhances the stability and efficacy of the nanoparticles by modifying their surface properties, leading to remarkably improved antibacterial activity against various human pathogens. These findings showcase the low-cost production medium, and extensive potential of pullulan not only for its intrinsic properties but also for its ability to significantly improve the performance of nanomaterials. This breakthrough opens doors to diverse applications in various fields.

Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant Enterococcus faecalis, the Causative Agent of Dental Abscess.

Background and Objectives: Enterococcus faecalis (E. faecalis) is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specific phages as an alternative method instead of classical treatments based on antibiotics to destroy multidrug-resistant E. faecalis bacteria for treating dental issues. Materials and Methods: In the current study, twenty-five bacterial isolates were obtained from infected dental specimens; only five had the ability to grow on bile esculin agar, and among these five, only two were described to be extensive multidrug-resistant isolates. Results: Two bacterial isolates, Enterococcus faecalis A.R.A.01 [ON797462.1] and Enterococcus faecalis A.R.A.02, were identified biochemically and through 16S rDNA, which were used as hosts for isolating specific phages. Two isolated phages were characterized through TEM imaging, which indicated that E. faecalis_phage-01 had a long and flexible tail, belonging to the family Siphoviridae, while E. faecalis_phage-02 had a contractile tail, belonging to the family Myoviridae. Genetically, two phages were identified through the PCR amplification and sequencing of the RNA ligase of Enterococcus phage vB_EfaS_HEf13, through which our phages shared 97.2% similarity with Enterococcus phage vB-EfaS-HEf13 based on BLAST analysis. Furthermore, through in silico analysis and annotations of the two phages' genomes, it was determined that a total of 69 open reading frames (ORFs) were found to be involved in various functions related to integration excision, replication recombination, repair, stability, and defense. In phage optimization, the two isolated phages exhibited a high specific host range with Enterococcus faecalis among six different bacterial hosts, where E. faecalis_phage-01 had a latent period of 30 min with 115.76 PFU/mL, while E. faecalis_phage-02 had a latent period of 25 min with 80.6 PFU/mL. They were also characterized with stability at wide ranges of pH (4-11) and temperature (10-60 °C), with a low cytotoxic effect on the oral epithelial cell line at different concentrations (1000-31.25 PFU/mL). Conclusions: The findings highlight the promise of phage therapy in dental medicine, offering a novel approach to combating antibiotic resistance and enhancing patient outcomes. Further research and clinical trials will be essential to fully understand the therapeutic potential and safety profile of these bacteriophages in human populations.

Unravelling the Antimicrobial, Antibiofilm, Suppressing Fibronectin Binding Protein A (fnba) and cna Virulence Genes, Anti-Inflammatory and Antioxidant Potential of Biosynthesized Solanum lycopersicum Silver Nanoparticles.

Background and Objectives: Urinary tract infections [UTIs] are considered the third most known risk of infection in human health around the world. There is increasing appreciation for the pathogenicity of Gram-positive and Gram-negative strains in UTIs, aside from fungal infection, as they have numerous virulence factors. Materials and Methods: In this study, fifty urine samples were collected from patients suffering from UTI. Among the isolates of UTI microbes, six isolates were described as MDR isolates after an antibiotic susceptibility test carried out using ten different antibiotics. An alternative treatment for microbial elimination involved the use of biosynthesized silver nanoparticles (AgNPs) derived from Solanum lycopersicum [S. cumin]. Results: The sizes and shapes of AgNPs were characterized through TEM imaging, which showed spherical particles in a size range of 35-80 nm, of which the average size was 53 nm. Additionally, the silver nanoparticles (AgNPs) demonstrated inhibitory activity against Staphylococcus aureus (OR648079), exhibiting a 31 mm zone of inhibition at a minimum inhibitory concentration (MIC) of 4 mg/mL and a minimum bactericidal concentration (MBC) of 8 mg/mL. This was followed by Aspergillus niger (OR648075), which showed a 30 mm inhibition zone at an MIC of 16 mg/mL and a minimum fungicidal concentration (MFC) of 32 mg/mL. Then, Enterococcus faecalis (OR648078), Klebsiella pneumoniae (OR648081), and Acinetobacter baumannii (OR648080) each displayed a 29 mm zone of inhibition at an MIC of 8 mg/mL and an MBC of 16 mg/mL. The least inhibition was observed against Candida auris (OR648076), with a 25 mm inhibition zone at an MIC of 16 mg/mL and an MFC of 32 mg/mL. Furthermore, AgNPs at different concentrations removed DPPH and H2O2 at an IC50 value of 13.54 µg/mL. Also, AgNPs at 3 mg/mL showed remarkable DNA fragmentation in all bacterial strains except Enterococcus faecalis. The phytochemical analysis showed the presence of different active organic components in the plant extract, which concluded that rutin was 88.3 mg/g, garlic acid was 70.4 mg/g, and tannic acid was 23.7 mg/g. Finally, AgNPs concentrations in the range of 3-6 mg/mL showed decreased expression of two of the fundamental genes necessary for biofilm formation within Staphylococcus aureus, fnbA (6 folds), and Cna (12.5 folds) when compared with the RecA gene, which decreased by one-fold when compared with the control sample. These two genes were submitted with NCBI accession numbers [OR682119] and [OR682118], respectively. Conclusions: The findings from this study indicate that biosynthesized AgNPs from Solanum lycopersicum exhibit promising antimicrobial and antioxidant properties against UTI pathogens, including strains resistant to multiple antibiotics. This suggests their potential as an effective alternative treatment for UTIs. Further research is warranted to fully understand the mechanisms of action and to explore the therapeutic applications of these nanoparticles in combating UTIs.

Lytic and Molecular Evidence of the Widespread Coriander Leaf Spot Disease Caused by Alternaria dauci.

Coriandrum sativum L. is a globally significant economic herb with medicinal and aromatic properties. While coriander leaf blight disease was previously confined to India and the USA, this study presents new evidence of its outbreak in Africa and the Middle East caused by Alternaria dauci. Infected leaves display irregular chlorotic to dark brown necrotic lesions along their edges, resulting in leaf discoloration, collapse, and eventual death. The disease also impacts inflorescences and seeds, significantly reducing seed quality. Koch's postulates confirmed the pathogenicity of the fungus through the re-isolation of A. dauci from artificially infected leaves, and its morphology aligns with typical A. dauci features. Notably, this study identified strong lytic activity (cellulase: 23.76 U, xylanase: 12.83 U, pectinase: 51.84 U, amylase: 9.12 U, and proteinase: 5.73 U), suggesting a correlation with pathogenicity. Molecular characterization using ITS (ON171224) and the specific Alt-a-1 gene (OR236142) supports the fungal morphology. This research provides the first comprehensive documentation of the pathological, lytic, and molecular evidence of A. dauci leaf blight disease on coriander. Future investigations should prioritize the development of resistant coriander varieties and sustainable disease management strategies, including the use of advanced molecular techniques for swift and accurate disease diagnosis to protect coriander from the devastating impact of A. dauci.

Thymus Vulgaris Oil Nanoemulsion: Synthesis, Characterization, Antimicrobial and Anticancer Activities.

Essential oil nanoemulsions have received much attention due to their biological activities. Thus, a thyme essential oil nanoemulsion (Th-nanoemulsion) was prepared using a safe and eco-friendly method. DLS and TEM were used to characterize the prepared Th-nanoemulsion. Our findings showed that the nanoemulsion was spherical and ranged in size from 20 to 55.2 nm. The micro-broth dilution experiment was used to evaluate the in vitro antibacterial activity of a Th-emulsion and the Th-nanoemulsion. The MIC50 values of the thymol nanoemulsion were 62.5 mg/mL against Escherichia coli and Klebsiella oxytoca, 250 mg/mL against Bacillus cereus, and 125 mg/mL against Staphylococcus aureus. Meanwhile, it emerged that the MIC50 values of thymol against four strains were not detected. Moreover, the Th-nanoemulsion exhibited promising antifungal activity toward A. brasiliensis and A. fumigatus, where inhibition zones and MIC50 were 20.5 ± 1.32 and 26.4 ± 1.34 mm, and 12.5 and 6.25 mg/mL, respectively. On the other hand, the Th-nanoemulsion displayed weak antifungal activity toward C. albicans where the inhibition zone was 12.0 ± 0.90 and MIC was 50 mg/mL. Also, the Th-emulsion exhibited antifungal activity, but lower than that of the Th-nanoemulsion, toward all the tested fungal strains, where MIC was in the range of 12.5-50 mg/mL. The in vitro anticancer effects of Taxol, Th-emulsion, and Th-nanoemulsion were evaluated using the standard MTT method against breast cancer (MCF-7) and hepatocellular carcinoma (HepG2). Additionally, the concentration of VEGFR-2 was measured, and the activities of caspase-8 (casp-8) and caspase-9 (casp-9) were evaluated. The cytotoxic effect was the most potent against the MCF-7 breast cancer cell line after the Th-nanoemulsion treatment (20.1 ± 0.85 µg/mL), and was 125.1 ± 5.29 µg/mL after the Th-emulsion treatment. The lowest half-maximal inhibitory concentration (IC50) value, 20.1 ± 0.85 µg/mL, was achieved when the MCF-7 cell line was treated with the Th-nanoemulsion. In addition, Th-nanoemulsion treatments on MCF-7 cells led to the highest elevations in casp-8 and casp-9 activities (0.66 ± 0.042 ng/mL and 17.8 ± 0.39 pg/mL, respectively) compared to those with Th-emulsion treatments. In comparison to that with the Th-emulsion (0.982 0.017 ng/mL), the VEGFR-2 concentration was lower with the Th-nanoemulsion treatment (0.672 ± 0.019ng/mL). In conclusion, the Th-nanoemulsion was successfully prepared and appeared in nanoform with a spherical shape according to DLS and TEM, and also exhibited antibacterial, antifungal, as well as anticancer activities.

Phytochemical analysis and insight into insecticidal and antifungal activities of Indian hawthorn leaf extract.

Fungicides or insecticides are popular means of controlling a variety of pathogens and insect pests; however, they can cause harmful effects on both human health and the environment. Different researchers have suggested using plant extracts, which have shown promise in managing fungi and insects. The purpose of this investigation was to explore the antifungal activities of an acetone extract made from the leaves of Indian Hawthorn (HAL) against phytopathogens that are known to harm maize crops, Fusarium verticillioides (OQ820154) and Rhizoctonia solani (OQ820155), and to evaluate the insecticidal property against Aphis gossypii Glover aphid. The HAL extract demonstrated significant antifungal activity against the two fungal pathogens tested, especially at the high dose of 2000 µg/mL. Laboratory tests on the LC20 of HAL extract (61.08 mg/L) versus buprofezin 25% WP (0.0051 mg/L) were achieved on A. gossypii Glover. HAL extract diminished the nymph's production over 72 h and their total reproductive rate. This extract was like buprofezin 25% WP in decreasing the daily reproductive rate, reproductive period, and mean survival percentage. Nevertheless, the newly-born nymphs of treated females with HAL extract attained the highest reduction in survival percentage at 46.00%. Equalized prolongations on the longevity of nymphs to 9.33, 8.33, and 7 days and the total life cycle to 15.00, 14.00, and 12.67 days were realized by HAL extract, buprofezin 25% WP, and the control, respectively. The olfactory choice test on the aphids showed the minimum attraction rate to HAL extract. The HPLC of HAL extract comprised an abundance of phenolic compounds (ferulic acid, gallic acid, 4-hydroxybenzoic acid, salicylic acid, ellagic acid, and pyrogallol), and the concentrations of these compounds vary widely, with salicylic acid being the most concentrated at 25.14 mg/mL. Among the flavonoids, epicatechin has the highest concentration at 11.69 mg/mL. The HAL extract GC-MS consists of various organic compounds, including sesquiterpenes, cyclopropenes, fatty acids, steroids, alcohols, ketones, esters, bufadienolides, opioids, and other organic compounds. The most abundant compounds in the sample are n-hexadecanoic acid (12.17%), followed by 5α, 7αH, 10α-eudesm-11-en-1α-ol (9.43%), and cis-13-octadecenoic acid (5.87%). Based on the findings, it can be inferred that the HAL extract may be a viable option for plants to combat both fungal and insect infestations. This presents an encouraging prospect for utilizing a natural and sustainable approach toward long-term pest management in plants.

Watermelon Rind Mediated Biosynthesis of Bimetallic Selenium-Silver Nanoparticles: Characterization, Antimicrobial and Anticancer Activities.

One of the most hazardous diseases that influences human health globally is microbial infection. Therefore, bimetallic nanoparticles have received much attention for controlling microbial infections in the current decade. In the present study, bimetallic selenium-silver nanoparticles (Se-Ag NPs) were effectively biosynthesized using watermelon rind WR extract through the green technique for the first time. UV-visible spectroscopy, transmission electron microscopy (TEM), and energy dispersive X-ray spectroscopy (EDX) methods were used to characterize the produced NPs. The results indicated that the bimetallic Se-Ag NPs had synergistic antimicrobial activity at low concentrations, which helped to reduce the toxicity of Ag NPs after the bimetallic Se-Ag NPs preparation and increase their great potential. Se-Ag NPs with sizes ranging from 18.3 nm to 49.6 nm were detected by TEM. Se-Ag NP surfaces were uniformly visible in the SEM picture. The cytotoxicity of bimetallic Se-Ag NPs was assessed against the Wi38 normal cell line to check their safety, where the IC50 was 168.42 µg/mL. The results showed that bimetallic Se-Ag NPs had antibacterial action against Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Klebsiella oxytoca, Bacillus subtilis, and Staphylococcus aureus with a minimum inhibitory concentration (MIC) of 12.5 to 50 µg/mL. Additionally, bimetallic Se-Ag NPs had promising anticancer activity toward the MCF7 cancerous cell line, where the IC50 was 21.6 µg/mL. In conclusion, bimetallic Se-Ag NPs were biosynthesized for the first time using WR extract, which had strong antibacterial, antifungal and anticancer properties.

Synergistic Anticandidal Effectiveness of Greenly Synthesized Zinc Oxide Nanoparticles with Antifungal Agents against Nosocomial Candidal Pathogens.

The high prevalence of fungal resistance to antifungal drugs necessitates finding new antifungal combinations to boost the antifungal bioactivity of these agents. Hence, the aim of the present investigation was to greenly synthesize zinc oxide nanoparticles (ZnO-NPs) using an aqueous leaf extract of Salvia officinalis and investigate their antifungal activity and synergistic efficiency with common antifungal agents. The biofabricated ZnO-NPs were characterized to detect their physicochemical properties. A disk diffusion assay was employed to investigate the antifungal effectiveness of the greenly synthesized ZnO-NPs and evaluate their synergistic patterns with common antifungal agents. The Candida tropicalis strain was detected to be the most susceptible strain to ZnO-NPs at both tested concentrations of 50 and 100 µg/disk, demonstrating relative suppressive zones of 19.68 ± 0.32 and 23.17 ± 0.45 mm, respectively. The minimum inhibitory concentration (MIC) of ZnO-NPs against the C. tropicalis strain was 40 µg/mL, whereas the minimum fungicidal concentration (MFC) was found to be 80 µg/mL. The highest synergistic efficiency of the biogenic ZnO-NPs with terbinafine antifungal agent was detected against the C. glabrata strain, whereas the highest synergistic efficiency was detected with fluconazole against the C. albicans strain, demonstrating relative increases in fold of inhibition area (IFA) values of 6.82 and 1.63, respectively. Moreover, potential synergistic efficiency was detected with the nystatin antifungal agent against the C. tropicalis strain with a relative IFA value of 1.06. The scanning electron microscopy (SEM) analysis affirmed the morphological deformations of candidal cells treated with the biosynthesized ZnO-NPs as the formation of abnormal infoldings of the cell wall and membranes and also the formation of pores in the cell wall and membranes, which might lead to the leakage of intracellular constituents. In conclusion, the potential synergistic efficiency of the biogenic ZnO-NPs with terbinafine, nystatin, and fluconazole against the tested candidal strains highlights the potential application of these combinations in formulating novel antifungal agents of high antimicrobial efficiency. The biogenic ZnO nanoparticles and antifungal drugs exhibit powerful synergistic efficiency, which highlights their prospective use in the formulation of efficient antimicrobial medications, including mouthwash, ointments, lotions, and creams for effective candidiasis treatment.

Antimicrobial, Antibiofilm, and Anticancer Activities of Syzygium aromaticum Essential Oil Nanoemulsion.

In the current study, clove oil nanoemulsion (CL-nanoemulsion) and emulsion (CL-emulsion) were prepared through an ecofriendly method. The prepared CL-nanoemulsion and CL-emulsion were characterized using dynamic light scattering (DLS) and a transmission electron microscope (TEM), where results illustrated that CL-nanoemulsion droplets were approximately 32.67 nm in size and spherical in shape, while CL-nanoemulsion droplets were approximately 225.8 nm with a spherical shape. The antibacterial activity of CL-nanoemulsion and CL-emulsion was carried out using a microbroth dilution method. Results revealed that the preferred CL-nanoemulsion had minimal MIC values between 0.31 and 5 mg/mL. The antibiofilm efficacy of CL-nanoemulsion against S. aureus significantly decreased the development of biofilm compared with CL-emulsion. Furthermore, results illustrated that CL-nanoemulsion showed antifungal activity significantly higher than CL-emulsion. Moreover, the prepared CL-nanoemulsion exhibited outstanding antifungal efficiency toward Candida albicans, Cryptococcus neoformans, Aspergillus brasiliensis, A. flavus, and A. fumigatus where MICs were 12.5, 3.12, 0.78, 1.56, and 1.56 mg/mL, respectively. Additionally, the prepared CL-nanoemulsion was analyzed for its antineoplastic effects through a modified MTT assay for evaluating apoptotic and cytotoxic effects using HepG2 and MCF-7 cell lines. MCF-7 breast cancer cells showed the lowest IC50 values (3.4-fold) in CL-nanoemulsion relative to that of CL-emulsion. Thus, CL-nanoemulsion induces apoptosis in breast cancer cells by inducing caspase-8 and -9 activity and suppressing VEGFR-2. In conclusion, the prepared CL-nanoemulsion had antibacterial, antifungal, and antibiofilm as well as anticancer properties, which can be used in different biomedical applications after extensive studies in vivo.

Zinc Oxide Nanoparticles Biosynthesized by Eriobotrya japonica Leaf Extract: Characterization, Insecticidal and Antibacterial Properties.

Zinc oxide nanoparticles (ZnO-NPs) have gained significant attention in nanotechnology due to their unique properties and potential applications in various fields, including insecticidal and antibacterial activities. The ZnO-NPs were biosynthesized by Eriobotrya japonica leaf extract and characterized by various techniques such as UV-visible (UV-vis) spectrophotometer, X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), and zeta potential analysis. The results of SEM revealed that NPs were irregular and spherical-shaped, with a diameter between 5 and 27 nm. Meanwhile, DLS supported that the measured size distributions were 202.8 and 94.7 nm at 11.1° and 90.0°, respectively, which supported the polydisperse nature of NPs, and the corresponding zeta potential was -20.4 mV. The insecticidal activity of the produced ZnO-NPs was determined against the adult stage of coleopteran pests, Sitophilus oryzae (Linnaeus) (Curculionidae) and Tribolium castaneum (Herbst) (Tenebrionidae). The LC50 values of ZnO-NPs against adults of S. oryzae and T. castaneum at 24 h of exposure were 7125.35 and 5642.65 µg/mL, respectively, whereas the LC90 values were 121,824.56 and 66,825.76 µg/mL, respectively. Moreover, the biosynthesized nanoparticles exhibited antibacterial activity against three potato bacterial pathogens, and the size of the inhibition zone was concentration-dependent. The data showed that the inhibition zone size increased with an increase in the concentration of nanoparticles for all bacterial isolates tested. The highest inhibition zone was observed for Ralstonia solanacearum at a concentration of 5 µg/mL, followed by Pectobacterium atrosepticum and P. carotovorum. Eventually, ZnO-NPs could be successfully used as an influential agent in pest management programs against stored-product pests and potato bacterial diseases.

Exploring Exogenous Indole-3-acetic Acid's Effect on the Growth and Biochemical Profiles of Synechocystis sp. PAK13 and Chlorella variabilis.

Microalgae have garnered scientific interest for their potential to produce bioactive compounds. However, the large-scale industrial utilization of microalgae faces challenges related to production costs and achieving optimal growth conditions. Thus, this study aimed to investigate the potential role of exogenous indole-3-acetic acid (IAA) application in improving the growth and production of bioactive metabolites in microalgae. To this end, the study employed different concentrations of exogenously administered IAA ranging from 0.36 µM to 5.69 µM to assess its influence on the growth and biochemical composition of Synechocystis and Chlorella. IAA exposure significantly increased IAA levels in both strains. Consequentially, improved biomass accumulation in parallel with increased total pigment content by approximately eleven-fold in both strains was observed. Furthermore, the application of IAA stimulated the accumulation of primary metabolites. Sugar levels were augmented, providing a carbon source that facilitated amino acid and fatty acid biosynthesis. As a result, amino acid levels were enhanced as well, leading to a 1.55-fold increase in total amino acid content in Synechocystis and a 1.42-fold increase in Chlorella. Total fatty acids content increased by 1.92-fold in Synechocystis and by 2.16-fold in Chlorella. Overall, the study demonstrated the effectiveness of exogenously adding IAA as a strategy for enhancing the accumulation of microalgae biomass and biomolecules. These findings contribute to the advancement of microalgae-based technologies, opening new avenues to produce economically important compounds derived from microalgae.

Technological Quality of Sugarcane Inoculated with Plant-Growth-Promoting Bacteria and Residual Effect of Phosphorus Rates.

Phosphate fertilization in highly weathered soils has been a major challenge for sugarcane production. The objective of this work was to evaluate the foliar levels of phosphorus (P) and nitrogen (N) and the technological quality and productivity of second ratoon cane as a function of inoculation with plant-growth-promoting bacteria (PGPBs) together with the residual effect of phosphate fertilization. The experiment was carried out at the research and extension farm of Ilha Solteira, state of São Paulo, Brazil. The experiment was designed in a randomized block with three replications in a 5 × 8 factorial scheme. The treatments consisted of five residual doses of phosphorus (0, 45, 90, 135 and 180 kg ha-1 of P2O5, 46% P) applied at planting from the source of triple superphosphate and eight inoculations from three species of PGPB (Azospirillum brasilense, Bacillus subtilis and Pseudomonas fluorescens), applied in single or co-inoculation at the base of stems of sugarcane variety RB92579. Inoculation with PGPBs influenced leaf N concentration, while inoculations with Pseudomonas fluorescens and combinations of bacteria together with the highest doses exerted a positive effect on leaf P concentration. Co-inoculation with A. brasilense + Pseudomonas fluorescens associated with a residual dose of 135 kg ha-1 of P2O5 increased stem productivity by 42%. Thus, it was concluded that inoculations with Pseudomonas fluorescens and their combinations are beneficial for the sugarcane crop, reducing phosphate fertilization and increasing productivity.

Sustainable biosynthesized bimetallic ZnO@SeO nanoparticles from pomegranate peel extracts: antibacterial, antifungal and anticancer activities.

Sustainable bimetallic nanoparticles (NPs) have attracted particular attention in the past decade. However, the efficiency and environmental concerns are associated with their synthesis and properties optimization. We report herein biosynthesis of bimetallic ZnO@SeO NPs based on green and ecofriendly methods using pomegranate peel extract (PPE). Pyrochemical ultraviolet-visible (UV-vis), Fourier-transform infrared (FTIR) and X-ray diffraction (XRD) spectroscopy as well as TEM and EDX supported successful synthesis. Antibacterial, antifungal, and cytotoxic activities were indicative of biological worth of sustainable bimetallic ZnO@SeO NPs, exhibiting antibacterial activity compared to monometallic ZnO and SeO NPs. The values of Minimum Inhibitory Concentration (MIC) of bimetallic ZnO@SeO NPs toward E. coli, P. aeruginosa, B. subtilis and S. aureus were 3.9, 15.62, 3.9 and 7.81 µg ml-1, respectively. Likewise, a promising antifungal activity against Candida albicans, Aspergillus flavus, A. niger and A. fumigatus was achieved (MICs: 31.25, 1.95, 15.62 and 15.62 µg ml-1, respectively). The cytotoxicity results suggest that bimetallic ZnO@SeO NPs are non-toxic and biomedically safe, evidenced by in vitro anticancer activity against human liver carcinoma (Hep-G2) cell line (with a half-maximal inhibitory concentration (IC50) > 71 µg ml-1). The bimetallic ZnO@SeO NPs successfully biosynthesized using PPE showed a high potential for biomedical engineering.

Wound Dressing Scaffold with High Anti-biofilm Performance Based on Ciprofloxacin-Loaded Chitosan-Hydrolyzed Starch Nanocomposite: In Vitro and In Vivo Study.

Today, the search for solutions to reduce wound infection and restore wound receptivity also reduces its side effects which are a difficult problem in medical science research. The greatest options for this purpose are hydrogel dressings since they are compatible with tissue and have an antibacterial effect on wound healing. Chronic wounds represent a significant burden on people and healthcare systems worldwide. Bacteria often enter such skin wounds, causing irritation and complicating the healing process. In addition, bacteria cause infection, which inhibits rejuvenation and the production of collagen. This study is aimed at developing novel chitosan (CS)-hydrolyzed starch nanocomposite (HS/Ch-NC) loaded with ciprofloxacin to enhance its skin retention and wound healing efficacy and anti-biofilm efficacy. Drug-loading on the (HS/Ch-NC) and encapsulation efficiency was 55.2% and 97.2%, respectively. The activity of HS-NC loaded with ciprofloxacin as anti-biofilm activity by 72% and 63% against Enterobacter aerogenes and Pseudomonas aeruginosa, respectively. The obtained (HS/Ch-NC) loaded with ciprofloxacin is a promising candidate for the development of improved bandage materials, as cell viability and proliferation was assessed using an SRB assay with half-maximal inhibitory concentrations (IC50) at 119.1 µg/ml. In vitro scratch wound healing assay revealed significant (p ≤ 0.05) acceleration in wound closure at 24 h enhanced by 56.04% 24-h and 100% 72-h post-exposure to (HS/Ch-NC) loaded ciprofloxacin, compared to the negative control. In vivo skin retention study revealed that (HS/Ch-NC)-loaded ciprofloxacin showed 3.65-fold higher retention, respectively, than ciprofloxacin. Thus, our study assumes that ciprofloxacin-loaded HS-NC is a potential delivery system for enhancing ciprofloxacin skin retention and wound healing activity.

Antiviral Activity of Biosynthesized Silver Nanoparticles from Pomegranate (Punica granatum L.) Peel Extract against Tobacco Mosaic Virus.

Tobacco mosaic virus (TMV) is a major pathogen affecting tomato plants worldwide. The efficacy of silver nanoparticles (Ag-NPs) mediated by Punica granatum biowaste peel extract in mitigating the negative impact of TMV infection on tomato growth and oxidative stress was investigated through scanning electron microscopy (SEM), transmission electron microscopy (TEM), UV-Visible (UV-Vis) spectrophotometer, X-ray Diffraction (XRD), dynamic light scattering (DLS), zeta potential, energy-dispersive X-ray spectroscopy (EDX), and Fourier-transform infrared spectra (FTIR). Results of SEM analysis of green Ag-NPs revealed the presence of condensed spherical or round NPs with diameters ranging between 61 and 97 nm. TEM confirmed the SEM results and showed round-shaped Ag-NPs with an average size of 33.37 ± 12.7 nm. The elemental analysis (EDX) of prepared Ag-NPs revealed the presence of elemental Ag as a major peak (64.43%) at 3-3.5 KeV. The FTIR revealed several functional groups on the prepared Ag-NPs, for which three treatment strategies for Ag-NP applications were evaluated in the greenhouse study and compared to inoculated TMV and control plants: pre-infection treatment (TB), post-infection treatment (TA), and dual treatment (TD). The results showed that the TD strategy is the most effective in improving tomato growth and reducing viral replication, whereas all Ag-NP treatments (TB, TA, and TD) were found to significantly increase expression of the pathogenesis-related (PR) genes PR-1 and PR-2, as well as polyphenolic compounds, HQT, and C4H genes compared to control plants. In contrast, the flavonoid content of tomato plants was not affected by the viral infection, while the phenolic content was significantly reduced in the TMV group. Furthermore, TMV infection led to a significant increase in oxidative stress markers MDA and H2O2, as well as a reduction in the enzymatic activity of the antioxidants PPO, SOD, and POX. Our results clearly showed that the application of Ag-NPs on TMV-infected plants reduces virus accumulation, delays viral replication in all treatments, and greatly enhances the expression of the CHS gene involved in flavonoid biosynthesis. Overall, these findings suggest that treatment with Ag-NPs may be an effective strategy to mitigate the negative impact of TMV infection on tomato plants.

Biosynthesis, Characterization, and Antifungal Activity of Novel Trimetallic Copper Oxide-Selenium-Zinc Oxide Nanoparticles against Some Mucorales Fungi.

Metal nanoparticles are assumed to be a new generation of biologically active materials. The integrations between more than one metal are synergetic multifunctional features. In the current study, trimetallic copper-selenium-zinc oxide nanoparticles (Tri-CSZ NPs) were successfully mycosynthesized using Aspergillus niger through an ecofriendly method for the first time. The biosynthesis of the particles was characterized using physiochemical and topographical analysis. The physiochemical analysis included Fourier transform infrared spectroscopy (FTIR), which affirmed that the biosynthesis of Tri-CSZ NPs relies on the functional groups of fungal filtrates. Additionally, the UV-visible and X-ray diffraction patterns were proposed for the formation of Tri-CSZ NPs; moreover, topography analysis confirmed that the micromorphology of the nanoparticles were similar to a stick, with ends having a tetragonal pyramid shape, and with an average nanosize of about 26.3 ± 5.4 nm. Cytotoxicity results reveled that the Tri-CSZ NPs have no cytotoxicity on the human normal cell line Wi 38 at low concentrations, where the IC50 was 521 µg/mL. Furthermore, the antifungal activity of the Tri-CSZ NPs was evaluated. The antifungal results revealed that the Tri-CSZ NPs have promising antifungal activity against Mucor racemosus, Rhizopus microsporus, Lichtheimia corymbifera, and Syncephalastrum racemosum, where the minimum inhibitory concentrations (MICs) were 1.95, 7.81, 62.5, and 3.9 µg/mL, and the minimum fungicidal concentrations (MFCs) were 250, 62.5, 125, and 1000 µg/mL, respectively. In conclusion, Tri-CSZ NPs were successfully mycosynthesized using A. niger, which have a promising antifungal activity against fungi causing mucormycosis.

Efficient Role of Endophytic Aspergillus terreus in Biocontrol of Rhizoctonia solani Causing Damping-off Disease of Phaseolus vulgaris and Vicia faba.

The wide spread of plant pathogens affects the whole world, threatening national food security. Various fungi including Rhizoctonia solani induce the fungal disease damping-off that negatively affects plant seedlings' growth. Recently, endophytic fungi are used as safe alternatives to chemical pesticides that harm plant and human health. Here, an endophytic Aspergillus terreus was isolated from Phaseolus vulgaris seeds to control damping-off diseases by improving the defense system in Phaseolus vulgaris and Vicia faba seedlings. Endophytic fungus was morphologically and genetically identified Aspergillus terreus, and it is deposited in GeneBank under accession OQ338187. A. terreus demonstrated antifungal efficacy against R. solani with an inhibition zone at 22.0 mm. Moreover, the minimum inhibitory concentrations (MIC) of ethyl acetate extract (EAE) of A. terreus were between 0.3125 and 0.625 mg/mL to inhibit R. solani growth. Precisely 58.34% of the Vicia faba plants survived when A. terreus was added compared with the untreated infected (16.67%). Similarly, Phaseolus vulgaris achieved 41.67% compared to the infected (8.33%). Both groups of treated infected plants showed reduced oxidative damage (reduced Malondialdehyde and hydrogen peroxide levels) as compared to untreated infected plants. Reduced oxidative damage was correlated with the increase in photosynthetic pigments and the antioxidant defense system including polyphenol oxidase, peroxidase, catalase, and superoxide dismutase enzyme activities. Overall, the endophytic A. terreus can be considered an effective tool to control the suppression of Rhizoctonia solani in legumes, especially Phaseolus vulgaris and Vicia faba, as an alternative to synthetic chemical pesticides that harm the environment and human health.

A Simple Formula of the Endophytic Trichoderma viride, a Case Study for the Management of Rhizoctonia solani on the Common Bean.

The utilization of beneficial endophytic microorganisms presents a promising and innovative strategy for attaining environmental sustainability and fostering development. The majority of microbial bioagents are unsuitable for preparation in a suitable granular formula, and few are prepared in complicated formulas. In this work, Trichoderma viride was simply prepared in a marketable granular formula to manage Rhizoctonia solani and improve common bean growth. The GC-MS analysis showed several antimicrobial compounds in the fungal filtrate. T. viride was able to suppress the phytopathogenic R. solani in the laboratory. The formula had up to 6 months of shelf-life viability. Under greenhouse conditions, the formula improved plant resistance against R. solani. Moreover, the vegetative plant growth and physiological performance (peroxidase, polyphenol, total phenols, phenylalanine ammonia-lyase, and photosynthetic pigments) of the common bean showed obvious promotion. The formula reduced the disease incidence by 82.68% and increased the yield by 69.28%. This work may be considered a step in the right direction for producing simple bioactive products on a large scale. Moreover, the study's findings suggest that this method can be considered a novel approach to enhancing plant growth and protection, in addition to reducing costs, improving handling and application, and maintaining fungal viability for enhancing plant growth and protecting against fungal infections.

Green Biosynthesis of Zinc Oxide Nanoparticles Using Pluchea indica Leaf Extract: Antimicrobial and Photocatalytic Activities.

Nanotechnology is playing a critical role in several essential technologies with nanoscale structures (nanoparticles) in areas of the environment and biomedicine. In this work, the leaf extract of Pluchea indica was utilized to biosynthesize zinc oxide nanoparticles (ZnONPs) for the first time and evaluated for antimicrobial and photocatalytic activities. Different experimental methods were used to characterize the biosynthesized ZnONPs. The biosynthesized ZnONPs showed maximum Ultraviolet-visible spectroscopy (UV-vis) absorbance at a wavelength of 360 nm. The X-Ray diffraction (XRD) pattern of the ZnONPs exhibits seven strong reflection peaks, and the average particle size was 21.9 nm. Fourier-transform infrared spectroscopy (FT-IR) spectrum analysis reveals the presence of functional groups that help in biofabrication. The existence of Zn and O was confirmed by the Energy-dispersive X-ray (EDX) spectrum and the morphology by SEM images. Antimicrobial studies showed that the biosynthesized ZnONPs have antimicrobial efficacy against Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Candida albicans and Cryptococcus neoformans where inhibition zones at concentration 1000 µg/mL were 21.83 ± 0.76, 13.0 ± 1.1, 14.9 ± 0.85, 24.26 ± 1.1, 17.0 ± 1.0, 20.67 ± 0.57 and 19.0 ± 1.0 mm respectively. Under both dark and sunlight irradiation, the photocatalytic activity of ZnONPs was evaluated towards the degradation of the thiazine dye (methylene blue-MB). Approximately 95% of the MB dye was broken down at pH 8 after 150 min of sunlight exposure. The aforementioned results, therefore, suggest that ZnONPs synthesized by implementing environmentally friendly techniques can be employed for a variety of environmental and biomedical applications.

Nematocidal and Bactericidal Activities of Green Synthesized Silver Nanoparticles Mediated by Ficus sycomorus Leaf Extract.

Nanoparticles effectively control most plant pathogens, although research has focused more on their antimicrobial than their nematocidal properties. This study synthesized silver nanoparticles (Ag-NPs) through a green biosynthesis method using an aqueous extract of Ficus sycomorus leaves (FS-Ag-NPs). The nanoparticles were characterized using SEM, TEM, EDX, zeta sizer, and FTIR. The TEM results showed that the synthesized NPs were nanoscale and had an average particle size of 33 ± 1 nm. The elemental silver signal at 3 keV confirmed the formation of Ag-NPs from an aqueous leaf extract of F. sycomorus. The FTIR analysis revealed the existence of several functional groups in the prepared Ag-NPs. The strong-broad band detected at 3430 cm-1 indicated the stretching vibration of -OH (hydroxyl) and -NH2 (amine) groups. The nematocidal activity of biosynthesized FS-Ag-NPs has been evaluated in vitro against the root-knot nematode Meloidogyne incognita at 24, 48, and 72 h. The FS-Ag-NPs at a 200 µg/mL concentration applied for 48 h showed the highest effectiveness, with 57.62% nematode mortality. Moreover, the biosynthesized FS-Ag-NPs were also tested for their antibacterial activity against Pectobacterium carotovorum, P. atrosepticum, and Ralstonia solanacearum. With the application of nanoparticles, the reduction in bacterial growth gradually increased. The most potent activity at all concentrations was found in R. solanacearum, with values of 14.00 ± 2.16, 17.33 ± 2.05, 19.00 ± 1.41, 24.00 ± 1.41, and 26.00 ± 2.83 at concentrations of 5, 10, 15, 20, and 25 µg/mL, respectively, when compared with the positive control (Amoxicillin 25 µg) with a value of 16.33 ± 0.94. At the same time, the nanoparticles showed the lowest reduction values against P. atrosepticum when compared to the control. This study is the first report on the nematocidal activity of Ag-NPs using F. sycomorus aqueous extract, which could be a recommended treatment for managing plant-parasitic nematodes due to its simplicity, stability, cost-effectiveness, and environmentally safe nature.