Controlled synthesis and computational analysis of gold nanostars for the treatment of Fusarium oxysporum.

Fusarium oxysporum (F. oxysporum) is linked to the widespread fusarium wilt in plants affecting the quality and yield of food crops. Management of fusarium wilt by synthetic fertilizers poses safety concerns. Safer-by-design nanomaterials synthesized with a greener approach can meet the needs of commercial antifungal drug resistance. Herein, a simple aqueous reduction method has been adopted for the synthesis of anisotropic gold nanostars (AuNSs) using quercetin-para aminobenzoic acid (QPABA) as both a reducing and stabilizing agent at room temperature for the treatment of F. oxysporum. QPABA was used to control the growth of Au3+ star-shaped nanoparticles at increasing concentrations in the ratio of 2 : 1 (QPABA : Au3+ ions) respectively. Transmission electron microscopy (TEM) analysis of the as-prepared gold nanoparticles confirmed the formation of nanostars with sizes of 40 ± 2 nm. The formation of anisotropic gold nanoparticles was evaluated by UV-vis characterizations which showed longitudinal surface plasmon modes at 540 and 800 nm. The gold nanoparticles exhibit excellent antifungal activity against F. oxysporum with the minimum inhibitory concentration (MIC) of 100 µg mL-1 using an agar well-diffusion assay. AuNSs proved to be efficacious in controlling F. oxysporum, as shown in the SEM analysis with a disintegrated cell membrane upon treatment. Computational analysis was performed to determine the specific binding sites on the QPABA ligand for gold ion interactions using the DFT B3LYP method, with a 6-31+G(d) basis set. Results showed that the interaction between Au3+ and QPABA at the 4 and 3 positions yielded the highest stability and formation of gold nanostars. The results suggest that the synthesized AuNSs act as a promising antifungal agent with great potential in treating frequent fungal infections that affect agricultural production.

Sugar-Lectin Interactions for Direct and Selective Detection of Escherichia coli Bacteria Using QCM Biosensor.

Pathogenic Escherichia coli (E. coli) remains a safety concern in the preservation and quality of green leafy vegetables. Sugar-lectin interactions provide a reliable, specific, and effective sensing platform for the detection of bacteria as compared to the tedious conventional plate counting technique. Herein, we present the synthesis of 4-(N-mannosyl) benzoic acid (4-NMBA) and 4-thiophenyl-N-mannose (4-TNM) via a two-step reductive amination for the detection of E. coli using a quartz crystal microbalance (QCM) biosensor. The 4-NMBA was synthesized with mannose and para-aminobenzoic (4-PBA), while the 4-TNM was synthesized with mannose and 4-aminophenyl disulfide (4-AHP) using water and acetic acid in a 1:1 ratio. The resultant structure of mannose derivatives (4-NMBA and 4-TNM) was characterized and confirmed using analytical tools, such as Mass Spectrometer, SEM, and FTIR. The choice of ligands (mannose derivatives) is ascribed to the specific recognition of mannose to the FimH lectin of the type 1 pilus of E. coli. Furthermore, the 4-PBA and 4-AHP conjugated to mannose increase the ligand affinity to FimH lectins. The setup of the QCM biosensor was composed of modification of the crystal surface and the covalent attachment of ligands for the detection of E. coli. The piezoelectric effect (frequency shift of the quartz) was proportional to the change in mass added to the gold crystal surface. Both the 4-NMBA- and 4-TNM-coated QCM sensors had a limit of detection of 3.7 CFU/mL and 6.6 CFU/mL with a sensitivity of 2.56 × 103 ng/mL and 8.99 × 10-5 ng/mL, respectively, within the dynamic range of 103 to 106 CFU/mL. This study demonstrates the application of ligand-coated QCM biosensors as a cost-effective, simple, and label-free technology for monitoring pathogenic bacteria via molecular interactions on crystal surfaces.

Ligand effect on controlling the synthesis of branched gold nanomaterials against fusarium wilt diseases.

The widespread wilt disease caused by Fusarium solani spp is a pressing problem affecting crop production and intensive farming. Strategic biocontrol of Fusarium solani spp using phytochemical mediated nano-materials is eco-friendly compared to harsh synthetic fungicides. The present study demonstrates the comparative dose effects of QPABA-derived branched gold nanomaterial (AuNF) and quercetin-mediated spherical gold nanoparticles (s-AuNPs) against Fusarium solani spp. Quercetin-para aminobenzoic acid (QPABA) was synthesized using reductive amination by reacting para-aminobenzoic acid with quercetin in an eco-friendly solvent at 25 °C. The structure elucidation was confirmed using 1H and 13C-NMR. TLC analysis showed that QPABA (R f = 0.628) was more polar in water than quercetin (R f = 0.714). The as-synthesized QPABA serves as a reducing and capping agent for the synthesis of gold nanoflowers (AuNFs) and gold nanostars (AuNSs). The UV-vis, XRD, and TEM confirmed the SPR peak of gold (550 nm) and gold element with a particle size distribution of 20-80 nm for the nanostars respectively. AuNFs exhibited a significant (P < 0.05) inhibitory effect against F. solani in a dose-dependent manner using Agar well diffusion. Nevertheless, spherical-AuNPs were not effective against F. solani. The inhibitory effect was influenced by the size, dose treatment, and particle shape. The minimum inhibitory concentration (MIC) value of AuNFs was 125.7 ± 0.22 µg mL-1. Our results indicate that AuNFs show considerable antifungal activity against F. solani as compared to spherical AuNPs. This study shows a greener synthesis of gold anisotropic nanostructures using QPAB, which holds promise for the treatment of fungal pathogens impacting agricultural productivity.

Colorimetric Detection of the SARS-CoV-2 Virus (COVID-19) in Artificial Saliva Using Polydiacetylene Paper Strips.

The spread and resurgence of the SARS-CoV-2 virus (COVID-19 disease) threatens human health and social relations. Prevention of COVID-19 disease partly relies on fabricating low-cost, point-of-care (POC) sensing technology that can rapidly and selectively detect the SARS-CoV-2 virus. We report a colorimetric, paper-based polydiacetylene (PDA) biosensor, designed to detect SARS-CoV-2 spike protein in artificial saliva. Analytical characterizations of the PDA sensor using NMR and FT-IR spectroscopy showed the correct structural elucidation of PCDA-NHS conjugation. The PDA sensor platform containing the N-Hydroxysuccinimide ester of 10, 12-pentacosadiynoic acid (PCDA-NHS) was divided into three experimental PCDA-NHS concentration groups of 10%, 20%, and 30% to optimize the performance of the sensor. The optimal PCDA-NHS molar concentration was determined to be 10%. The PDA sensor works by a color change from blue to red as its colorimetric output when the immobilized antibody binds to the SARS-CoV-2 spike protein in saliva samples. Our results showed that the PDA sensing platform was able to rapidly and qualitatively detect the SARS-CoV-2 spike protein within the concentration range of 1 to 100 ng/mL after four hours of incubation. Further investigation of pH and temperature showed minimal influence on the PDA sensor for the detection of COVID-19 disease. After exposure to the SARS-CoV-2 spike protein, smartphone images of the PDA sensor were used to assess the sensor output by using the red chromatic shift (RCS) of the signal response. These results indicate the potential and practical use of this PDA sensor design for the rapid, colorimetric detection of COVID-19 disease in developing countries with limited access to medical testing.

Size and Shape-Dependent Antimicrobial Activities of Silver and Gold Nanoparticles: A Model Study as Potential Fungicides.

Plant-based pathogenic microbes hinder the yield and quality of food production. Plant diseases have caused an increase in food costs due to crop destruction. There is a need to develop novel methods that can target and mitigate pathogenic microbes. This study focuses on investigating the effects of luteolin tetraphosphate derived silver nanoparticles (LTP-AgNPs) and gold nanoparticles (LTP-AuNPs) as a therapeutic agent on the growth and expression of plant-based bacteria and fungi. In this study, the silver and gold nanoparticles were synthesized at room temperature using luteolin tetraphosphate (LTP) as the reducing and capping agents. The synthesis of LTP-AgNPs and LTP-AuNP was characterized by Transmission Electron Microscopy (TEM) and size distribution. The TEM images of both LTP-AgNPs and LTP-AuNPs showed different sizes and shapes (spherical, quasi-spherical, and cuboidal). The antimicrobial test was conducted using fungi: Aspergillus nidulans, Trichaptum biforme, Penicillium italicum, Fusarium oxysporum, and Colletotrichum gloeosporioides, while the class of bacteria employed include Pseudomonas aeruginosa, Aeromonas hydrophila, Escherichia coli, and Citrobacter freundii as Gram (-) bacteria, and Listeria monocytogenes and Staphylococcus epidermidis as Gram (+) bacterium. The antifungal study demonstrated the selective size and shape-dependent capabilities in which smaller sized spherical (9 nm) and quasi-spherical (21 nm) AgNPs exhibited 100% inhibition of the tested fungi and bacteria. The LTP-AgNPs exhibited a higher antimicrobial activity than LTP-AuNPs. We have demonstrated that smaller sized AgNPs showed excellent inhibition of A. nidulans growth compared to the larger size nanoparticles. These results suggest that LTP-AuNP and LTP-AgNPs could be used to address the detection and remediation of pathogenic fungi, respectively.

Antimicrobial Activity of a New Class of Phosphorylated and Modified Flavonoids.

The surge of resistant food pathogens is a major threat worldwide. Previous research conducted on phytochemicals has shown their antibacterial activity against pathogenic bacteria. The design of antimicrobial agents to curb pathogenic disease remains a challenge demanding critical attention. Flavonoids such as apigenin and quercetin were evaluated against Gram-positive and Gram-negative bacteria. The results indicated that the antibacterial activity of each flavonoid occurred at a different minimum inhibitory concentration. However, the antimicrobial activity results of the modified flavonoids were also reported, and it was observed that the Gram-positive bacteria were more susceptible in comparison to the Gram-negative bacteria. The cell wall structure of the Gram-positive and Gram-negative bacteria could be the main reason for the bacteria susceptibility. Modified flavonoids could be used as a suitable alternative antimicrobial agent for the treatment of infectious diseases. Our results indicated 100% inhibition of Listeria monocytogenes, Pseudomonas aeruginosa, and Aeromonas hydrophila with modified flavonoids.

Selective Sensing and Imaging of Penicillium italicum Spores and Hyphae Using Carbohydrate-Lectin Interactions.

The blue-green mold Penicillium italicum is among the most problematic post-harvest plant infections limiting the integrity of citrus and many other crops during storage and transportation, but there is no sensor for its on-site or field detection. We hereby, for the first time, report the development of novel biomolecular sensor for assessing the presence of P. italicum spores and hyphae using carbohydrate-lectin recognitions. Two approaches were developed: (i) lateral tests using standalone poly(amic) acid (PAA) membranes and glass surfaces and (ii) quantitative tests on 96-well polystyrene plates and paper electrodes. In both cases, the surfaces were functionalized with novel derivatized sugar based ligands while staining was performed with gold nanoparticles. Both approaches provided strong signals for 104 spores/mL of P. italicum isolated from experimentally infected lemons as the lowest-reliable concentration. The 96-well plate-based gave the most sensitive detection with a 4 × 102 spores/mL limit of detection, a linear dynamic range between 2.9 × 103 and 6.02 × 104 spores/mL ( R2 = 0.9939) and standard deviation of less than 5% for five replicate measurements. The selectivity of the ligands was tested against Trichaptum biforme, Glomerulla cingulata ( Colletotrichum gloeosporioides), and Aspergillus nidulans fungi species. The highest selectivity was obtained using the sugar-based gold-nanoparticles toward both the spores and the hyphae of P. italicum. The advanced specificity was provided by the utilized sugar ligands employed in the synthesis of gold nanoparticles and was independent from size and shapes of the AuNPs. Accuracy of the sensor response showed dramatic dependence on the sample preparation. In the case of 5-10 min centrifugation at 600 rpm, the spores can be isolated free from hyphae and conidiophore, for which spiked recovery was up to 95% (std ±4). In contrast, for gravity-based precipitation of hyphae, the spiked recovery was 88% (std 11).

Flavonoid-derived anisotropic silver nanoparticles inhibit growth and change the expression of virulence genes in Escherichia coli SM10.

We hereby present a novel greener and ecofriendly synthesis of anisotropic silver nanoparticles (AgNPs) using water soluble quercetin diphosphate (QDP). QDP was employed as a reducing, capping and stabilizing agent at room temperature without any extraneous reagents. The purpose of this study was to determine the effects of modified quercetin pentaphosphate silver nanoparticles (QPP-AgNPs) and quercetin diphosphate derived silver nanoparticles (QDP-AgNPs) on microbial growth and expressions of virulence-related genes in Escherichia coli SM10. The gene expression analysis was carried out for 12 genes which are related to virulence and stress in E. coli SM10, namely: RpoD, RpoS, ibpB, clpB, uspA, fliC, fimH, fimF, kdpE, artJ, hyaA, and gyrA. Results showed that QDP-AgNPs reduced the swarming motility by 98% which correlated with the reduction in the expression of FliC flagellar gene. A simultaneous increase in the expression of the fimbrial genes FimH and FimF that are related to motility was recorded. In contrast, treatment of the microbes with QPP-AgNPs resulted in 90% of the swarming motility at different patterns compared to QDP-AgNPs treatment for the gene expressions of motility elements. The study revealed that QDP-AgNPs up-regulated the stress related RpoD and ibpB expressions, while QPP-AgNPs up-regulated the stress related RpoS and uspA gene expressions. However, both QDP-AgNPs and QPP-AgNPs up-regulated kpdE, artJ and gry at different levels. QDP-AgNPs were also tested for their antibacterial and antifungal activities, which showed µmolar cidal activity. The growth kinetics of both Gram (-) and Gram (+) bacteria were strongly altered by QDP-AgNPs activity. Energy dispersive absorption spectroscopy (EDS) studies revealed that silver ions and/or the nanoparticles themselves transferred into bacterial cells. To the best of our knowledge, this is the first report ofstudying the genetic and kinetic response of bacteria to modified quercetin phosphate mediated silver nanoparticles and we hereby report that the molecules used to synthesize AgNPs bring about a strong effect on AgNPs manipulatory activity on the tested 12-genes.