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Reasons for Performing Study: Incisional drainage and suppuration occurs commonly following exploratory laparotomy; any technique with the potential to reduce the incidence of this complication warrants investigation., Objectives: To determine if abandoning the use of subcutaneous sutures in laparotomy wound closure is safe and whether it reduces the risk of suppuration., Methods: A randomised controlled study was carried out at 2 referral hospitals in the UK, involving 309 horses undergoing exploratory laparotomy; 150 horses underwent '2-layer' closure without a subcutaneous suture, while 159 underwent conventional '3-layer' closure. Information regarding degree of oedema and gross types of discharge was recorded daily; suppuration was defined as discharge of pus. Telephone follow-up was carried out 30 days after hospital discharge to identify those complications occurring after that date and, thereafter, every 3 months. The influence of closure method on risk of wound suppuration was assessed by Chi-squared analysis and by logistic regression. Time to suppuration was modelled using a Cox proportional hazards model., Results: No catastrophic failures of 2-layer closures were recorded. Prevalence of suppuration was not significantly different, being 18.7% and 23.9% for 2- and 3-layer closures, respectively (OR = 1.37, 0.79-2.37, P = 0.263)., Conclusions and Potential Relevance: This study found no significant difference in prevalence or rate of wound suppuration in 2-layer closures compared to conventional 3-layer closure. Two-layer closure is recommended as a safe alternative means of achieving ventral midline abdominal closure in horses.