Objective: To assess the effectiveness of a protocol for prevention of surgical site infection (SSI) in colorectal surgery. Patients and methods: Evaluation of 2 cohorts of patients undergoing colon and rectal surgery in a tertiary public hospital: A historical cohort (2008-2011) and a prospective one (after the implementation of the program in 2012). The main measures established were: Adequacy of preoperative antimicrobial prophylaxis, maintaining patient normothermia and appropriate glove change during the intervention. Comparability of the two cohorts was determined by a bivariate analysis of age, sex, NNIS index, ASA index, surgical time, perioperative transfusion, diagnosis, diabetes and renal failure. Results: We assessed 342 patients (256 underwent colon surgery and 86 rectal surgery), divided into 2 cohorts: prior period (218), and post-implementation period (124). The cumulative incidence of SSI in the first cohort was 27.5% (95% CI, 21.6-33.4), and in the post-intervention cohort 16.9% (95% CI, 10.3-23.5, P=.03). Postoperative mortality was 9.2% (95% CI, 5.4-13) in the first cohort and 3.2% (95% CI, 0.1-6.3) in the post-intervention cohort (P=.04). The inadequacy of prophylaxis decreased from 37.4% (95% CI, 30.4-44.6) to 18.9% (95% CI, 11.9-26.1) (P=.001). Conclusion: A significant decrease in the frequency of SSI, post-surgical mortality and inadequate antimicrobial prophylaxis is verified after the implementation of a protocol in colorectal surgery (C) 2014 AEC. Published by Elsevier Espana, S.L.U. All rights reserved.