Objectives: To determine whether wearing colored indicator gloves affects perforation detection rate and to identify risk factors for glove perforation during veterinary orthopedic surgery. Study Design: Prospective randomized controlled trial. Sample population: 574 double pairs of gloves worn during 300 orthopedic surgical procedures (total 2296 surgical gloves). Methods: Primary and assistant surgeons double gloved for all orthopedic surgical procedures. Type of inner glove (standard (n=180) or colored indicator (n=180)) was randomized for the first 360 double pairs of gloves worn by surgeons during 180 procedures. Perforations detected by the surgeon were recorded and gloves changed if requested. For a further 120 procedures, indicator gloves were used exclusively. All gloves were leak tested following surgery to identify perforations. Association between potential risk factors and perforation were explored using multivariate logistical regression analysis. Results: Glove perforations occurred during 43% of surgeries with a mean of 2.3 holes per surgery. Where an outer perforation occurred, inner gloves were intact in 63%. Intra-operative perforation detection was improved when colored indicator gloves were worn (83% sensitivity) versus standard gloves (34% sensitivity, p<0.0001). Independent risk factors for perforation identified were the placement of plates and/or screws (p=0.001, OR=2.4, 95% CI=1.4-4.0), placement of an external skeletal fixator (p=0.002, OR=7.0, 95% CI=2.1-23.8), the use of orthopedic wire (p=0.011, OR=2.4, 95% CI=1.2-4.7) and the primary surgeon being board-certified (p=0.016, OR=1.9, 95% CI=1.1-3.1). Conclusions: Colored indicator gloves increase surgeon recognition of glove perforations enabling gloves to be changed promptly if perforation occurs.