Objective: To document natural bacterial flora on the ventral aspect of the equine abdomen, to compare 2 preparation techniques, and to identify potential risk factors that may contribute to incisional drainage., Design: Prospective study., Animals: 53 horses undergoing exploratory celiotomy., Procedure: Group-1 horses (n = 26) were prepared with povidone-iodine and alcohol. Group-2 horses (27) were prepared with a film-forming iodophor complex. Numbers of bacterial colony-forming units (CFU) were measured before and after surgical scrub, following skin closure, and after recovery from general anesthesia. Swab specimens to identify normal skin bacterial flora and potential pathogens were obtained by swabbing a 4 x 4-cm area. Variables that might affect incisional drainage were also investigated., Results: For both techniques, there was a significant reduction in bacterial numbers after skin preparation. Incisional drainage was observed in 14 (26%) horses (8 group-1 and 6 group-2 horses). Preexisting dermatitis, poor intraoperative drape adherence, high number of bacterial CFU obtained after recovery from anesthesia, and high number of CFU obtained from the surgery room environment were the main risk factors associated with subsequent incisional drainage. Bacillus spp, nonhemolytic Staphylococcus spp, Micrococcus spp, Corynebacterium spp, Streptomyces spp, other nonenteric genera, and nonhemolytic Streptococcus spp were the most common isolates obtained before surgical scrub., Conclusions and Clinical Relevance: Both skin preparation techniques were equally effective in reducing numbers of bacterial CFU by 99%, and a significant difference was not found in incisional drainage rate between groups. Protection of the wound during recovery from anesthesia and the immediate postoperative period may reduce incisional drainage after abdominal surgery in horses.