To investigate the therapeutic effect of 275 nm wavelength ultraviolet C (UV-C) light for treatment of bacterial keratitis in canine corneas using an affordable, broadly available modified handheld device. UV-C therapy (UVCT) was evaluated in two experiments: invitro using triplicates of three bacterial genera (Staphylococcus, Streptococcus, Pseudomonas spp., and a mix of all species) where the UVCT was performed at a distance of 10, 15, and 20 mm with 1 or 2 doses (4 h apart) for 5, 15, or 30 s; exvivo model where healthy canine corneal buttons were inoculated superficially and deep (330 μm) with the same bacterial isolates and treated at a 10 mm distance for 15 s with one dose of 22.5 mJ/cm2. Fluorescent marker (STYO9-PI) was used to label (green = live bacteria, red = dead bacteria), and confocal microscopy was used to image the bacteria. In vitro results showed all plates treated with UVCT had 100% bactericidal effect for all isolates with single dose of 15 s at 10 mm distance or two doses, 4 h apart at 15 mm and was ineffective with single dose at 15-20 mm. The exvivo results confirmed a significant decrease in bacterial load for all isolates on samples inoculated superficially but were inconclusive for intrastromal ones. UVCT confirmed the therapeutic potential for all tested isolates, for both invitro and exvivo experiments using a single exposure of 15 s. While safety studies are underway, clinical trials are warranted.