Abstract Background Hand hygiene is critical for infection prevention in surgery. While surgical scrubbing and sterile gloving are routine, the risk of microbial regrowth beneath gloves during prolonged procedures is poorly studied. This study assessed microbial proliferation on hands after gloving and its implications for intraoperative sterility. Methods A prospective observational study was conducted in the operating theatre. Surgical staff underwent standard scrubbing before donning sterile gloves. Hand swabs were taken immediately after gloving and at fixed time intervals (up to 6 h). Samples were cultured using standard microbiological techniques and colony growth was recorded. Results No growth was observed immediately after scrubbing and gloving. Microorganisms were first detected after 4 h of continuous glove use, with significant proliferation by 6 h. Predominant isolates were skin commensals, including Staphylococcus species. This indicates that hands are no longer sterile beneath gloves during prolonged surgery, and may pose a risk of contamination when gloves are changed or removed. Conclusions Hands do not remain sterile beneath gloves during prolonged surgery. Significant microbial growth after 6 h increases the risk of contamination during intraoperative glove changes, particularly in procedures involving implants or meshes. Moreover, if gloves are removed at the end of surgery and hands are not re-washed, subsequent contact with wounds or theatre surfaces poses a direct infection risk. Reinforcing strict handwashing after surgery and before regloving is essential to minimise surgical site infection.
Agrawal et al. (2026) studied this question.