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Based onpreliminary results of 23 animal experiments, the aim of this clinical trial was to asses autonomic nerve function by intraoperative nerve stimulation (INS) with simultaneous observation of internal anal sphincter (IAS) activity and bladder manometry.Three patients with low rectal tumours underwent nerve-preserving total mesorectal excision. INS was performed under observation of amplified IAS neuromonitoring signals and bladder manometry. Urogenital and anorectal function were assessed with residual urine volume (RUV), International Prostate Symptom Score (IPSS), International Index of Erectile Function (IIEF), Wexner-Vaizey-Score, Cleveland-Clinic-Incontince-Score (CCIS) and Bowel Function Instrument (MSKCC) questionnaires. Patients were studiedpreoperatively, postoperatively (10 day) and will be followed up at 3, 6, 12 months after surgery.In all patients INS resulted in increasing IAS signals ( (0.38 μV (95 % CI: 0.28; 0.48) vs 2.98 μV (95 % CI: 1.89; 4.07)), (P < 0.001). Median increase in intravesicalpressure was 3.9 cm H2O (3.0; 4.8). Preoperative RUV was in median 5.4 ml (0.0; 5.7), IPSS 3.7 (±2.1), IIEF 17.7 (±14.7), Wexner-Vaizey-Score 1.0 (±1.0), CCIS 2.0 (±2.6). Postoperative RUV was in median 6.0 ml (0.0; 30.0), IPSS 3.7 (±2.1).INS with observation of IAS neuromonitoring signals represents a novel method for identification and verification of pelvic autonomic nerve function (BMBF, Grant number 01EZ0722).