This study aimed to investigate the suture length to wound length ratio (SL:WL) in an in vitro model of abdominal wall closure. Effects of the surgeon's experience level on the SL:WL ratio were evaluated, hypothesizing that small animal surgeons do not spontaneously apply SL:WL ratios equal to or larger than 4:1.Three groups of surgeons with varying levels of experience performed 4 simple continuous sutures before (3 sutures) and after (1 suture) being educated on principles of the SL:WL ratio. All sutures were evaluated for their gaping, number of stitches, stitch intervals, tissue bite size and suture length.No significant differences in suture parameters or SL:WL ratios were found among the 3 groups, and 60.5% of control sutures and 77.0% of test sutures had SL:WL ratios above 4:1. There was a significant improvement in the mean ratio after the information was provided (p = 0.003). Overall, the SL:WL ratios ranged from 1.54:1 to 6.81:1, with 36.3% falling between 4:1 and 5:1 (5.17 mm mean stitch interval, 5.52 mm mean tissue bite size). A significant negative correlation was observed between the SL:WL ratio and the stitch interval to tissue bite ratio (r = -0.886). Forty-nine of 120 sutures fulfilled the current recommendations for abdominal wall closure with a mean SL:WL ratio of 4.1:1.A SL:WL ratio larger than 4:1 was achieved in 60% of the control sutures and in 77% of test sutures. Additional animal studies are necessary to evaluate the SL/WL ratio in small animal surgery.