Potential Antimicrobial and Antibiofilm Properties of Copper Oxide Nanoparticles: Time-Kill Kinetic Essay and Ultrastructure of Pathogenic Bacterial Cells

Mycosynthesis of nanoparticle (NP) production is a potential ecofriendly technology for large scale production. In the present study, copper oxide nanoparticles (CuONPs) have been synthesized from the live cell filtrate of the fungus Penicillium chrysogenum. The created CuONPs were characterized via several techniques, namely Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), and energy-dispersive X-ray spectroscopy (EDX). Furthermore, the biosynthesized CuONPs were performed against biofilm forming Klebsiella oxytoca ATCC 51,983, Escherichia coli ATCC 35,218, Staphylococcus aureus ATCC 25,923, and Bacillus cereus ATCC 11,778. The anti-bacterial activity result was shown with the zone of inhibition determined to be 14 ± 0.31 mm, 16 ± 0.53 mm, 11 ± 0.57 mm, and 10 ± 0.57 mm respectively. Klebsiella oxytoca and Escherichia coli were more susceptible to CuONPs with minimal inhibitory concentration (MIC) values 6.25 and 3.12 µg/mL, respectively, while for Staphylococcus aureus and Bacillus cereus, MIC value was 12.5 and 25 μg/mL, respectively. The minimum biofilm inhibition concentration (MBIC) result was more evident, that the CuONPs have excellent anti-biofilm activity at sub-MIC levels reducing biofilm formation by 49% and 59% against Klebsiella oxytoca and Escherichia coli, while the results indicated that the MBIC of CuONPs on Bacillus cereus and Staphylococcus aureus was higher than 200 μg/mL and 256 μg/mL, respectively, suggesting that these CuONPs could not inhibit mature formatted biofilm of Bacillus cereus and Staphylococcus aureus in vitro. Overall, all the results were clearly confirmed that the CuONPs have excellent anti-biofilm ability against Klebsiella oxytoca and Escherichia coli. The prepared CuONPs offer a smart approach for biomedical therapy of resistant microorganisms because of its promoted antimicrobial action, but only for specified purposes.