The goal behind this work is to prepare, characterize, and study the antimicrobial behavior of zirconia (ZrO 2 ) nanoparticles (NPs). Various techniques, such as X-ray diffraction (XRD), were used for studying the mineralogical structure and crystal size. The microstructure and chemical composition of the prepared particles were analyzed using a scanning electron microscope attached with an energy-dispersive X-ray analysis (EDAX) unit. The antagonistic ability against several Gramnegative and Gram-positive bacteria, including Salmonella paratyphi, Pseudomonas aeruginosa, Alcaligenes aquatilis, Escherichia coli, Streptococcus pneumoniae, and Staphylococcus aureus, was assessed using the well diffusion method. The results of XRD and scanning electron microscopy (SEM) analyses revealed that the prepared material exhibited the phase of zirconium nanoparticles with particle sizes ranging between 40 and 75 nm. The antimicrobial test results demonstrated that the inhibitory effect increased with the increase of concentration. The inhibitory effect was more pronounced against Gram-positive bacteria, as indicated by the larger size of the inhibitory zone. At a 9% dimethyl sulfoxide (DMSO) concentration, the inhibitory zone had a diameter of 3.50 mm for S. aureus compared to a diameter of 3.40 mm for S. pneumoniae. The use of zirconium oxide nanoparticles reduced the diameter of the inhibitory zone when tested against S. aureus at a 3% DMSO concentration (0.50 mm diameter) and against S. pneumoniae (0.40 mm diameter). Zirconia nanoparticles were also evaluated for their antifungal activity against several species, including Aspergillus niger, Aspergillus f lavus, and Penicillium sp. The size of the inhibitory zone indicated the susceptibility of microorganisms to nanozirconium oxide, resulting in a stronger inhibition of Penicillium sp. at a 100% DMSO concentration (4.50 mm diameter) compared to A. niger and A. f lavus (3.00 mm diameter). The results for Penicillium sp. at a 3% DMSO concentration showed a diameter of the inhibitory zone of 0.90 mm, while for A. niger and A. flavus, the diameter was 0.80 mm. Thus, our findings demonstrate that the zirconium oxide nanoparticles possess the capability to reduce the inhibition zone effectively for both bacterial and fungal activities.