Background: Perforations of surgical gloves are common and increase with the duration of glove wear. Skin flora, re-grown after pre-operative disinfection of the hands, may contaminate a surgical site. An antimicrobial surgical glove with chlorhexidine on its inner surface has been developed. We hypothesized that by suppressing the re-growth of skin flora during the complete course of a surgical procedure, antimicrobial gloves may reduce the risk of surgical site contamination in the event of an intra-operative glove breach. Methods: We conducted a randomized, double-blind, single-center trial, to measure any differences in the bacterial skin populations of surgeons' hands during surgical procedures done with antimicrobial and non-antimicrobial surgical gloves [ISRCTN71391952]. In this study, 25 pairs of gloves were retrieved from 14 surgeons who donned them randomly on their dominant or non-dominant hand. The number of bacteria retrieved from glove fluid was measured and expressed as colony forming units (CFU)/mL. Results: The median cfu/mL of antimicrobial gloves was 0.00 (LQ: 0.00 CFU/mL; UQ: 0.00 cfu/mL), with a mean log10 cfu/mL=0.02 (range: 0.00-0.30). The median CFU/mL of non-antimicrobial gloves was 54.00 (LQ: 3.00 cfu/mL; UQ: 100.00 cfu/mL) with a mean log10 CFU/mL=1.32 (range: 0.00-2.39). After a mean operating time of 112 min, the difference in the log10 CFU/mL was 1.30 (p<0.001). Conclusions: A new antimicrobial surgical glove suppressed surgeons' hand flora during operative procedures. In the event of a glove breach, the use of such a glove may have the potential to prevent bacterial contamination of a sterile surgical site, thereby decreasing the risk of surgical site infection (SSI) and increasing patient safety. Further clinical studies are needed to confirm this concept.