ABSTRACT
Bacterial biofilms are associated with antibiotic resistance and account for â¼80% of all bacterial infections. In this study, we explored novel nanomaterials for combating bacteria and their biofilms. Artemisinin nano-copper (ANC) was synthesized using a green synthesis strategy, and its shape, size, structure, elemental composition, chemical valence, zeta potential, and conductivity were characterized using transmission electron microscopy, X-ray diffractometer, X-ray photoelectron spectroscopy, zeta potential, and dynamic light scattering. The results showed that ANC was successfully synthesized utilizing a liquid phase chemical reduction method using chitosan as a modified protectant and l-ascorbic acid as a green reducing agent. The stability of ANC was evaluated using dynamic light scattering. The results showed that the particle size of ANC at different concentrations was comparable to that of the original solution after 7 days of storage, and there was no significant change in the polydispersity index (P > 0.05). The antibacterial effects of ANC on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were determined by disc diffusion and broth dilution methods. The results demonstrated that ANC inhibited and killed E. coli and S. aureus. The effect of ANC on bacterial biofilms was investigated using crystal violet staining, scanning electron microscopy, laser confocal microscopy, and quantitative polymerase chain reaction. The results showed that ANC treatment was able to destroy bacterial biofilms and downregulate biofilm- and virulence-related genes in E. coli (HlyA, gyrA, and F17) and S. aureus (cna, PVL, ClfA, and femB). Green-synthesized ANC possesses excellent antibiofilm properties and is expected to exhibit antibacterial and antibiofilm properties.
Subject(s)
Anti-Bacterial Agents , Artemisinins , Biofilms , Copper , Escherichia coli , Green Chemistry Technology , Staphylococcus aureus , Biofilms/drug effects , Copper/chemistry , Copper/pharmacology , Escherichia coli/drug effects , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology , Green Chemistry Technology/methods , Artemisinins/pharmacology , Artemisinins/chemistry , Microbial Sensitivity Tests , Nanostructures/chemistry , Metal Nanoparticles/chemistryABSTRACT
BACKGROUND: Bacterial panicle blight, incited by Burkholderia glumae, has impacted rice production globally. Despite its significance, knowledge about the disease and the virulence pattern of the causal agent is very limited. Bacterial panicle blight is a major challenge in the rice-growing belts of North-western India, resulting in yield reduction. However, the management of B. glumae has become a challenge due to the lack of proper management strategies. METHODOLOGY AND RESULTS: Twenty-one BG strains have been characterized using the 16S rRNA and the gyrB gene-based sequence approach in the present study. The gyrB gene-based phylogenetic analysis resulted in geographic region-specific clustering of the BG isolates. The virulence screening of twenty-one BG strains by inoculating the pathogenic bacterial suspension of 1 × 10-8 cfu/ml at the booting stage (55 DAT) revealed the variation in the disease severity and the grain yield of rice plants. The most virulent BG1 strain resulted in the highest disease incidence (82.11%) and lowest grain yield (11.12 g/plant), and the least virulent BG10 strain resulted in lowest disease incidence of 18.94% and highest grain yield (24.62 g/plant). In vitro evaluation of various biocontrol agents and nano copper at different concentrations by agar well diffusion method revealed that nano copper at 1000 mg/L inhibited the colony growth of B. glumae. Under net house conditions, nano copper at 1000 mg/L reduced the disease severity to 21.23% and increased the grain yield by 20.91% (31.76 g per plant) compared to the positive control (COC 0.25% + streptomycin 200 ppm). Remarkably, pre-inoculation with nano copper at 1000 mg/L followed by challenge inoculation with B. glumae enhanced the activity of enzymatic antioxidants viz., Phenyl ammonia-lyase (PAL), Polyphenol oxidase (PPO) and Peroxidase (POX) and non-enzymatic antioxidant phenol. Additionally, we observed a substantial transcript level upregulation of six defense-related genes to several folds viz., OsPR2, OsPR5, OsWRKY71, OsPAL1, OsAPX1, and OsPPO1 in comparison to the pathogen control and healthy control. CONCLUSIONS: Overall, our study provides valuable insights into the potential and practical application of nano copper for the mitigation of bacterial panicle blight, offering promising prospects for commercial utilization in disease management.
Subject(s)
Burkholderia , Oryza , Oryza/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Burkholderia/genetics , Antioxidants , Copper , Edible GrainABSTRACT
Objective@#To investigate the antibacterial properties, biocompatibility and mechanical properties of Cu-ZnO-loaded dental veneering porcelain to provide an experimental basis for the development of new dental veneering porcelain. @*Methods@#Cu-ZnO nanoparticles were added to IPS E.max Ceram for restorative veneer porcelain at different mass percentages of 0 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, and 6 wt% using ball milling in ceramic powder. A cylindrical specimen with a diameter of 20 mm and a thickness of 2 mm was prepared by high-temperature sintering. Scanning electron microscopy was used to observe the surface morphologies of nano-Cu-ZnO and the specimens. The antibacterial effect of Escherichia coli (E. coli) was quantitatively studied by the plate colony counting method. The CCK-8 method was used to evaluate in vitro the cytotoxicity of the tested piece to mouse fibroblasts (L929). Live and dead cells were observed by fluorescence microscopy. The mechanical properties of modified IPS E. Max Ceram veneering porcelain were tested by a three-point bending strength test. @* Results @# Under the scanning electron microscope, Cu-ZnO appears with a block-like structure and can be seen dispersed in the veneering porcelain. When the nano Cu-ZnO loading was 1 wt%, 2 wt%, 3 wt%, and 4 wt%, the antibacterial rates of the specimens were 24.85%, 67.94%, 96.92%, and 99.99%, respectively, and the difference between the experimental groups and the control group was statistically significant (F = 23.308,P = 0.001). The relative growth rate of each group was greater than 80% after coculture with mouse fibroblast cells (L929) for 1 day and 3 days, and there was no significant difference between the groups. The morphology of L929 cells was normal after coculture for 24 hours. With the increase in the Cu-ZnO concentration, the flexural strength of the specimen exhibited an increasing trend followed by a decreasing trend. The bending strength of the specimen loaded with 3 wt% nano Cu-ZnO reached the maximum value (84.728 ± 6.82) MPa, and there was no statistically significant difference between groups (F = 0.633,P = 0.702).@*Conclusion@#The antibacterial rate of IPS E. max Ceram veneering porcelain loaded with 3 wt% nano Cu-ZnO was more than 96% against E. coli after high-temperature sintering at 750 ℃. The bending strength reached the maximum (84.728 ± 6.82) MPa, and there was no obvious cytotoxicity.
ABSTRACT
Aim To establish the rapidly detecting method of glucose concentration in brain microdialysate.Method Using high-performance liquid chromatography (HPLC) with electrochemical detector (ED) and Nano-Copper screen-printed electrode (NCSE100)-Flow injection analysis the glucose concentration in brain microdialysate were detected.Result The related coefficient (r) of glucose standard curve was 0.9995. The variation coefficient (CV) within a day was 4%~7%. The striatum extracellular glucose concentration in rats was(2.4?0.8) mmol?L-1. These results indicated that our method was reliable.Conclusion Using HPLC with ED, NCSE100, and flow injection analysis the glucose concentration in brain microdialysate were detected, which has certain applied worth.