Fine particle bombarding (FPB) is typically utilized to modify metal surfaces by bombarding them with fine particles at high-speed. The diameters of the particles range from several to tens of micrometers. FPB forms fine microscale concavities and convexities on a surface. As FPB-treated surfaces are widely used in the food industry, the influence of bacteria on their surface must be considered. In this study, we examined the antibacterial activity of microscale rough surfaces formed by FPB. We applied FPB to a stainless-steel surface and evaluated the antibacterial effect of FPB-treated surfaces based on JIS Z 2801 (a modified test method from ISO 22196:2007). Our results indicated that the FPB-treated surfaces (FPB-1 (avg. pitch: 0.72 µm) and FPB-2 (avg. pitch: 3.56 µm)) exhibited antibacterial activity both against Escherichia coli and Staphylococcus aureus. Graphical Abstract