The aim of this study was to evaluate the effectiveness of different UVC treatments, alone or in combination with ultra-high pressure homogenization (UHPH) on Bacillus subtilis spores in milk. Spores of B. subtilis (CECT4002) were inoculated in whole and skim milk to an initial concentration about 6 log CFU/mL. Milk was subjected to different ultraviolet radiation treatments at 254 nm (UVC) using a concentric tubular reactor in a dose ranging from 10 to 160 J/mL. Different number of passes were used to adjust the final dose received by the matrix. In general, increasing the number of passes (defined as number of entries to the tunnel-NET) increased the inactivation of spores of B. subtilis. The best lethality results (above 4 Log CFU/mL) were obtained by applying doses from 100 J/mL with several NET. When the same doses were achieved with a single pass lethality in most cases did not exceed 1 log CFU/mL. Increasing the NET also increased the likelihood for the spores to remain longer in the effective distance from the UVC source, estimated as 0.02 mm for whole milk and 0.06 mm for skim milk. Combination of UHPH and UVC did not clearly increase the efficiency of a single UVC treatment, and a lower lethality was even observed in some cases. UHPH treatments increased the turbidity and absorption coefficient (254 nm) of both whole and skim milk.