In this research, cerium oxide nanoparticles were synthesized using orange peel extract via a hydrothermal method. An equal ratio of orange peel extract to cerium nitrate salt led to the formation of cerium hydroxide carbonate, whereas a 1 : 10 ratio formed cerium oxide. The hydrothermal treatment was conducted for durations of 5 and 25 hours. Scanning electron microscopy (SEM) images revealed that the hydrothermal samples treated for 5 hours exhibited significant agglomeration in both extract to salt ratios after heat treatment. X-ray diffraction patterns confirmed that all samples were converted into cerium oxide after heating at 500 °C for 3 hours. Based on XRD and SEM results, three cerium oxide samples, including those synthesized through the 25 hours hydrothermal process with a 1 : 10 ratio and the 25 hours hydrothermal process with both ratios and subsequent heat treatment, were selected for further investigation. Fourier transform infrared (FT-IR) analysis revealed more adsorption of the functional groups of orange peel extract on the surface of the as-synthesized sample. Moreover, the heat-treated sample with a 1 : 10 ratio, initially cerium oxide, displayed a higher amount of surface functional groups than the one with a 1 : 1 ratio which was initially cerium hydroxide carbonate. The antibacterial activities of the samples were determined using the colony count method. Activities of all samples against Gram-negative bacteria are in the range of 91.5-93.2% with a negligible difference, whereas the as-synthesized sample exhibited a superior activity of 96.6 ± 1.8% against Gram-positive bacteria compared to the other two heat-treated samples. The 87.3% antioxidant activity of the as-synthesized sample significantly surpassed that of the other two samples, as evaluated by the DPPH radical scavenging method.