Hiba oil is widely used as a natural antibacterial agent, but it is highly volatile; therefore, it is important to preserve its effectiveness. In this study, we aimed to develop a material with high and sustained antibacterial activity as well as low environmental load by retaining Hiba oil in bamboo powder or cellulose nanofibers (CNF) and compositing it with cellulose acetate butyrate (CAB) to prepare a film. The antibacterial sustainability of the film was evaluated by the qualitative halo method, and the sustainability of the antibacterial activity was evaluated by the shaking method of a quantitative test. In the halo method, the antibacterial activity of the film without the retainer was preserved for 14 days, while that of the film containing 8 wt% bamboo powder was sustained for 35 days. In the shaking method, the film without the retainer had sufficient antibacterial activity only at the time of sample preparation (0 days), but the film with hydrophilic and hydrophobic CNF had sufficient antibacterial activity up to 7 days. These results confirm that the addition of a biomass-derived retainer helps maintain the antimicrobial effect of Hiba oil.