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The necessity for routine histopathologic evaluation of hemorrhoidectomy specimens considered free of suspicious areas after careful visual and manual inspection remains controversial.The purpose of this work was to prospectively study the prevalence of anal intraepithelial neoplasia in macroscopically normal operative specimens.From October 2005 to September 2010, all hemorrhoidectomy and fissurectomy specimens were sent for routine histopathologic analysis.This study was conducted at a tertiary referral center.The primary outcome measured was the histopathologic examination of surgical samples.Among the specimens from 2997 procedures, routine histopathologic evaluation found anal intraepithelial neoplasia in 97 patients (3.2%), despite the fact that visual and manual inspection had determined that the specimens were free of any suspected anal intraepithelial neoplasia or human papillomavirus-related lesion. The pathological diagnoses for these macroscopically normal specimens were AIN1 in 22 (23%) patients, AIN2 in 48 (49%) patients and AIN3 in 27 (28%) patients, making the prevalence of high-grade and low-grade disease 2.5% (anal intraepithelial neoplasia 2/3) and 0.7% (anal intraepithelial neoplasia 1).This study was limited by being a single-center study.This prospective single-center study demonstrated that the prevalence of infraclinical anal intraepithelial neoplasia in macroscopically normal hemorrhoidectomy and fissurectomy specimens is not negligible (3.2% with 2.5% high-grade disease).