This study was designed to assess the efficacy of using povidone-iodine (PVI) 10% and levofloxacin 0.5% about one hour before cataract surgery in minimizing aqueous contamination compared with using PVI 10%. The intervention group received three drops of mixed povidone-iodine and levofloxacin (PVI/Levofloxacin) solution one hour before surgery. After making the corneal incision in cataract surgery, 0.1 cc aqueous humor sample was taken and sent for culture. We compared the intervention group and control group using t-test and chi-square test in SPSS 25(Chicago IL, USA). 147 eyes of 147 patients were analyzed. The intervention and control groups consisted of 72 and 75 patients, respectively. The Control group showed significantly more positive cultures (16 cultures: 22.2%). the intervention group had four positive cultures (5.3%). (P-value = 0.005). Topical levofloxacin 0.5% and povidone-iodine 10% mixture showed promising results in reducing the number of bacteria in the aqueous humor.