A simple and rapid microwave assisted method of green synthesis of silver nanoparticles (AgNPs) was developed using aqueous leaf extract of Eucalyptus globulus(ELE), and their antibacterial and antibiofilm potential investigated. With this aim, the aqueous solutions of ELE and AgNO3(1 mM) were mixed (1:4 v/v), and microwave irradiated at 2450 Mhz, for 30 sec. The instant color change of the ELE-AgNO3 mixture from pale yellow to dark brown indicated ELE-AgNPs synthesis. The intensity of peak at 428 nm in UV-Vis spectra, due to the surface plasmon resonance of AgNPs, varied with the amount of ELE, AgNO3 concentration, pH and time of incubation. The biosynthesized ELE-AgNPs were characterized by UV-visible spectroscopy, XRD, TEM, SEM-EDX, FTIR and TGA analyses. The size of ELE-AgNPs was determined to be in range of 1.9–4.3 nm and 5-25 nm, with and without microwave treatment, respectively. SEM exhibited the capping of AgNPs with the ELE constituents, and validated by FTIR analysis. The FTIR data revealed the presence of plant organic constituents and metabolites bound to ELE-AgNPs, which contributes for their stability. The antimicrobial activity of ELE-AgNPs was assessed by growth and biofilm inhibition of extended spectrum β-lactamase (ESBL) producing Pseudomonas aeruginosa, Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) clinical bacterial isolates. The results demonstrated that S. aureus were more sensitive to ELE-AgNPs than E. coli and P. aeruginosa. MRSA exhibited higher sensitive than MSSA, whereas P. aeruginosa were more sensitive than E. coli to ELE-AgNPs treatment. Also, significant (83 ± 3% and 84 ± 5%) biofilm inhibition was observed in case of S. aureus and P. aeruginosa, respectively. The results elucidated environmentally friendly, economical and quick method for production of colloidal bio-functionalized ELE-AgNPs, for effectual clinical applications, as broad spectrum antibacterial agents and biofilm inhibitors.
The unregulated discharge of nanoparticles (NPs) from various nanotechnology industries into the environment is expected to alter the composition and physiological functions of soil microbiota. Considering this knowledge gap, the impact of five NPs (Ag, ZnO, CuO, Al2O3, and TiO2) differing in size and morphology on growth behavior and physiological activity of Azotobacter chroococcum, Bacillus thuringiensis, Pseudomonas mosselii, and Sinorhizobium meliloti were investigated. Various biochemical and microscopic approaches were adopted. Interestingly, all bacterial strains were found sensitive to Ag-NPs and ZnO-NPs but showed tolerance toward CuO, Al2O3, and TiO2-NPs. The loss of cellular respiration due to NPs was coupled with a reduction in population size. ZnO-NPs at 387.5 μg mL–1 had a maximum inhibitory impact on A. chroococcum and reduced its population by 72%. Under Ag-NP stress, the reduction in IAA secretion by bacterial strains followed the order S. meliloti (74%) > P. mosselii (63%) > A. chroococcum (49%). The surface of bacterial cells had small- or large-sized aggregates of NPs. Also, numerous gaps, pits, fragmented, and disorganized cell envelopes were visible. Additionally, a treated cell surface appeared corrugated with depressions and alteration in cell length and a strong heterogeneity was noticed under atomic force microscopy (AFM). For instance, NPs induced cell roughness for P. mosselii followed the order 12.6 nm (control) > 58 nm (Ag-NPs) > 41 nm (ZnO-NPs). TEM analysis showed aberrant morphology, cracking, and disruption of the cell envelope with extracellular electron-dense materials. Increased permeability of the inner cell membrane caused cell death and lowered EPS production. Ag-NPs and ZnO-NPs also disrupted the surface adhering ability of bacteria, which varied with time and concentration of NPs. Conclusively, a plausible mechanism of NP toxicity to bacteria has been proposed to understand the mechanistic basis of ecological interaction between NPs and resourceful bacteria. These results also emphasize to develop strategies for the safe disposal of NPs.
Chemically synthesized copper oxide nanoparticles (CuONPs) involve the generation of toxic products, which narrowed its biological application. Hence, we have developed a one-pot, green method for CuONP production employing the leaf extract of Cymbopogon citratus (CLE). Gas chromatography-mass spectrometry (GC-MS) analysis confirmed the capping of CuONPs by CLE esters (CLE-CuONPs). Fourier-transform infrared (FTIR) showed phenolics, sugars, and proteins mediated nucleation and stability of CLE-CuONPs. X-ray diffraction (XRD) and transmission electron microscopy (TEM) revealed CLE-CuONPs between 11.4 to 14.5 nm. Staphylococcus aureus-1 (MRSA-1), Staphylococcus aureus-2 (MSSA-2) exposed to CLE-CuONPs (1500 µg/mL) showed 51.4%, 32.41% survival, while Escherichia coli-336 (E. coli-336) exposed to 1000 µg/mL CLE-CuONPs showed 45.27% survival. Scanning electron microscopy (SEM) of CLE-CuONPs treated E. coli-336, MSSA-2 and MRSA-1 showed morphological deformations. The biofilm production by E. coli-336 and MRSA-1 also declined to 33.0 ± 3.2% and 49.0 ± 3.1% at 2000 µg/mL of CLE-CuONPs. Atomic absorption spectroscopy (AAS) showed 22.80 ± 2.6%, 19.2 ± 4.2%, and 16.2 ± 3.6% accumulation of Cu2+ in E. coli-336, MSSA-2, and MRSA-1. Overall, the data exhibited excellent antibacterial and antibiofilm efficacies of esters functionalized CLE-CuONPs, indicating its putative application as a novel nano-antibiotic against multi drug resistance (MDR) pathogenic clinical isolates.
1 July 2019 in Materials Science and Engineering: C