The major goal of this research was to determine how well NiO nanoparticles (NPs) inhibited the growth of harmful bacteria that were separated from biofilm layers that had developed inside of medical equipment used on a regular basis in hospitals and dental offices. Chia seed extract was previously used in the preparation and physical characterization of NiO nanoparticles. Thirty isolates of Pseudomonas aeruginosa (n=16) and Staphylococcus aureus (n=14) were found in separate swab samples taken from dental office bottles, syringe tubes, spittoons, ventilation machine tubes, and emergency room bottles. Using conventional bacteriological techniques, moist swab samples were inoculated and identified on culture medium, and subsequently antibiotic susceptibility was assessed in accordance with NCCLs. Using two different techniques—well diffusion and microdilution, respectively—the antibacterial activity of NiO nanoparticles against contaminated bacteria was assessed both qualitatively and quantitatively. The antibacterial susceptibility of suggested antibiotics was determined using a Muller Hinton agar (MHA) plate. Antibacterial susceptibility of bacterial isolates revealed considerable resistance to the antibiotics utilized in the current investigation. According to these results, NiO nanoparticles showed the greatest effectiveness when used at a concentration of 30 mg/mL in well diffusion, and the mean inhibition zones for P. aeruginosa and S. aureus isolates ranged from 16 to 20 mm, as opposed to 16 mm for the positive control (ofloxacin disk). In this work, the inhibition zone diameters of NiO nanoparticles were within the region of high susceptibility of positive control (16 mm/ Ofloxacin disk breakpoint). However, the minimum inhibitory concentration (MIC) of NiO nanoparticles was 1.875 mg/ml for 57.1% of S. aureus isolates and 0.468 mg/ml for 31.2% of P. aeruginosa isolates, with no isolates inhibited at high doses. The MIC findings revealed that the lowest concentration of NiO nanoparticles had in vitro efficiency and proved to be a good antibacterial agent against contaminated bacteria.
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