Abstract
Four novel Pseudomonas strains with record resistance to copper (Cu(2+)) previously isolated from ecologically diverse samples (P. lactis UKR1, P. panacis UKR2, P. veronii UKR3, and P. veronii UKR4) were tested against sonochemically synthesised copper-oxide (I) (Cu(2)O) and copper-oxide (II) (CuO) nanoparticles (NPs). Nanomaterials characterisation by X-ray diffractometry (XRD), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and High-Resolution Transmission Electron Microscopy (HRTEM) confirmed the synthesis of CuO and Cu(2)O NPs. CuO NPs exhibited better performance in inhibiting bacterial growth due to their heightened capacity to induce oxidative stress. The greater stability and geometrical shape of CuO NPs were disclosed as important features associated with bacterial cell toxicity. SEM and TEM images confirmed that both NPs caused membrane disruption, altered cell morphology, and pronounced membrane vesiculation, a distinctive feature of bacteria dealing with stressor factors. Finally, Cu(2)O and CuO NPs effectively decreased the biofilm-forming ability of the Cu(2+)-resistant UKR strains as well as degraded pre-established biofilm, matching NPs' antimicrobial performance. Despite the similarities in the mechanisms of action revealed by both NPs, distinctive behaviours were also detected for the different species of wild-type Pseudomonas analysed. In summary, these findings underscore the efficacy of nanotechnology-driven strategies for combating metal tolerance in bacteria.