The aim of this study was to determine the effect of chlorhexidine gluconate on microbial growth in patients undergoing debridement for diabetic foot. Material and Methods: The study had a randomized, controlled experimental design. The study sample included a total of 60 patients. Of 60 patients, 30 were randomly assigned into the experimental group and 30 were randomly assigned into the control group. In the experimental group, the surgical site was washed with chlorhexidine gluconate 0.05% before and after debridement in addition to routine cleaning with povidone iodine before debridement. In the control group, all the steps of the procedure performed in the experimental group were followed except for using chlorhexidine gluconate 0.05%. Results: The experimental and control groups were found to be similar in terms of descriptive characteristics and diabetesrelated features. There was no difference in the presence of microorganisms before debridement between the experimental and control groups, but fewer species were isolated from the experimental group after debridement. The mean number of isolated species in the experimental group was 1.66±0.60 and 0.60±0.67 before and after debridement respectively. The mean number of isolated species in the control group was 1.06±0.58 and 1.10±0.75 before and after debridement respectively. Conclusion: It can be concluded that chlorhexidine gluconate and povidone iodine used before debridement are effective in reduction of the number of microbial species after debridement in patients with diabetic foot.