: To evaluate the in vivo efficacy of rose bengal (RB)-mediated photodynamic antimicrobial therapy (PDAT) for treatment of keratitis (AK).: An animal (rabbit) AK model was successfully achieved via intrastromal inoculation of a suspension of cells and trophozoites. Prior to RB-PDAT (pre-treatment, day-5), the severity of the induced corneal infection was graded numerically for epithelial defects, stromal edema, neovascularity, and stromal opacity/infiltration. The right eyes of rabbits (n=18) were divided equally into three groups (n=6/group): control (no treatment); 0.1% RB+518 nm irradiation (5.4 J/cm); and 0.2% RB+518 nm irradiation (5.4 J/cm). On post-treatment day-5, animals were euthanized, after which corneal buttons were excised and submitted for real-time polymerase chain reaction (RT-PCR) analysis.: Post-treatment clinical scores of the 0.1 and 0.2% RB groups indicated significant improvement compared to control group scores (pre-treatment clinical scores; 5.17±0.98, 7.50±0.62, and 6.17±0.70 and post-treatment clinical scores; 4.50±0.56, (=0.043), 3.50±0.99 (=0.039), 6.83±1.66 (=0.34), respectively). RT-PCR analysis revealed that the mean cycle threshold (Ct) values were significantly higher in treated-group corneas compared to control-group corneas, with no significant differences between treated-groups (Mean Ct values; 34.33, 34.5, and 29.67 for 0.1 and 0.2% RB, and control groups). There was a statistically significant negative correlation between post-treatment clinical scores and Ct values (=-0.474, -value 0.047).: Our results demonstrate that RB-PDAT is effective in decreasing the parasitic load and clinical severity of AK.