Bismuth oxide nanoparticle Bi2O3NPs has a wide range of applications and less adverse effects than conventional radio sensitizers. In this work, Bi2O3NPs (D1, D2) were successfully synthesized by using the biosynthesis method with varying bismuth salts, bismuth sulfate Bi2(SO4)3 (D1) or bismuth nitrate. Penta hydrate Bi(NO3)3.5H2O (D2) with NaOH with beta-vulgaris extract. The Bi2O3NPs properties were characterized by different spectroscopic methods to determine Bi2O3NPs structure, nature of bonds, size of nanoparticle, element phase, presence, crystallinity and morphology. The existence of the Bi2O3 band was verified by the FT-IR. The Bi2O3 NPs revealed an absorption peak in the UV-visible spectrum, with energy gap Eg = 3.80eV. The X-ray pattern (D1) matching that of card of COD File-No-96-152-6459 indicating the presence of homogeneous Bi2O3NPs, Scaning Electron Microscopy (SEM) displayed shaped monoclinic phase with average diameter 30.28 nm. The size, structure and composition of synthetic Bi2O3 Nps were determined using the (EDX) pattern. The XRD pattern (D2) corresponds to JCPDS File No. 27-50. The SEM of D2 showed crystalline rhomobohedrral phase with average diameter 34.89 nm. The EDX for both (D1, D2) samples reveals an aggregation of thin sheet cluster. The antibacterial activity of Bi2O3NPs from (D1, D2) was tested against (G-) Escherichia coli and (G+) staphylococcus aureus. All of these clinical pathogens were examined for antifungal activity against Candida albicans fungus, and the results were compared with the standard medication. The adsorption experiment was successfully conducted on the following metal ions (M+2 = Co, Ni and Cu), where the results proved removal simultaneously from water using Bi2O3NPs (D1, D2) based on the affinity of three metal ions and Bi2O3 NPs surface shape. The removal efficiencies of mixed (M+2 = Co, Ni and Cu) ions for D1 were 89.68%, 85.56% and 94.5%. The removal efficiencies for D2 were 93.3%, 87.7% and 88.54%, respectively.