Purpose: We analyzed the technical feasibility, physiological influence, and safety of transvaginal (TV) and transrectal (TR) natural orifice translumenal endoscopic surgery (NOTES) nephrectomy compared with conventional laparoscopic surgery in a porcine survival model. Methods: Fifteen female pigs (32.8-37.2 kg) were randomly assigned to undergo TV NOTES (n=5), TR NOTES (n=5), or conventional laparoscopic nephrectomy (n=5). Postoperatively, all animals were observed for global health status during the recovery from anesthesia. Variable laboratory parameters and inflammatory cytokines were compared among the groups during the entire experimental period. Postmortem examination was performed 1 week after operation for assessment of abdominal complications and cultures for microorganisms. Results: All experiments were completed successfully without insertion of an extra port or conversion to laparoscopic or open surgery. Although mean operative times were longer in TV and TR NOTES groups than in the conventional laparoscopy group (61 vs 84 vs 24 min, P<0.001), there were no signs of visceral injury or peritonitis on postmortem examination. None of the laboratory parameters, including white blood cell count, tumor necrosis factor-alpha, interleukin (IL)-1, and IL-6 differed among the groups during the entire experimental period. Conclusions: TV and TR NOTES nephrectomy could be completed successfully with similar physiologic influence to those of conventional laparoscopic nephrectomy in a porcine survival model.