Objective To explore the bacterial translocation of mesenteric lymph nodes (MLNs) of the ileum and the spectrum of bacteria in patients with small bowel obstruction.Methods Total 84 patients were divided into study group (with small bowel obstruction) and control group (without small bowel obstruction). MLNs were obtained under sterile conditions intraoperatively, and which were processed for culture of aerobic and anaerobic organisms. The rate of bacterial translocation and postoperative infection were compared between two groups and the species of bacterial translocation was identified. Results The bacterial translocation rate in the study group was higher than that in the control group 〔57.1% (24/42) versus 16.7% (7/42),χ2=14.775, P<0.01〕. Escherichia coil was the most commonly bacteria (20). Emergency surgery and age over 70 years were associated with bacterial translocation (P<0.05). Postoperative infection complications rate in the bacterial translocation patients was higher than that in the patients without bacterial translocation 〔29.0% (9/31) versus 3.8% (2/53),χ2=10.965,P<0.05〕. Conclusions Bacterial translocation to MLNs occurres more frequently in patients with small bowel obstruction,non-elective surgery, and elderly.