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Yttrium oxide (Y₂O₃) is a promising rare-earth compound that is rarely studied. This study reports on the synthesis, structural, morphological, optical and antimicrobial properties of Y₂O₃ nanoparticles (NPs) synthesized using Agathosma betulina leaf extract. XRD pattern indicated that the crystalline phase was achieved for the sample calcined at $500^{\circ }$C with body-centered cubic symmetry. SEM and TEM images of Y₂O₃ nanopowder showed the nearly spherical morphology of agglomerated NPs with an average diameter of 13nm. Optical properties revealed that the bandgap of Y₂O₃ NPs ($\sim 5.67$eV) has been increased compared with the bulk Y₂O₃ ($\sim 5.5$eV). Moreover, the antimicrobial activity was also evaluated against gram-positive and gram-negative bacteria of clinical interest. Results demonstrated that the synthesized Y₂O₃ NPs exhibited higher bacteriostatic effects against gram-negative bacterial strains due to their thin cell wall structure. These results show that the synthesized Y₂O₃ NPs are suitable for antibacterial control systems and medicines.