Introduction: Neurogenic, inflammatory and pharmacological alterations during surgery contribute to the pathophysiology of postoperative ileus (POI). During the early onset of POI spinal afferent activation seems to occur which may trigger a reflex inhibition of intestinal motility perpetuating POI. However, the role of central vagal afferents in POI is still unknown. We, therefore, aimed to explore central vagal afferent nerve activation in the early development of POI.Methods: Under enflurane anesthesia, C57BL/6 mice underwent laparotomy followed by sham treatment or standardized small bowel manipulation to induce POI. Then, after 1h, 3h or 9h, the brain and jejunum was removed and fixed. Fos-immunoreactivity was determined for neuronal activation in the vagal nucleus of the solitary tract (nTS) of the brainstem and leucocyte infiltration in the intestinal muscularis by myeloperoxidase stains (each subgroup n=6). Data were analyzed by two-way ANOVA. Results: The number of Fos-positive neurons in the nTS did not differ 1h and 3h after induction of POI, but was significantly increased to 38±1, 48±3, 43±3 during ileus compared to 17±1, 18±2, 23±3 in sham controls (Bregma minus 13.3, 13.8, 14.3mm) 9h after small bowel manipulation (all P<0.05). The intestinal muscularis contained more leucocytes during ileus compared to controls 9h after small bowel manipulation (P<0.05). No change occurred 1h and 3h after induction of POI. Conclusions: While reflex inhibition of intestinal motility via spinal afferents seems to occur within the first hour after small bowel manipulation, central vagal afferents were activated after the recruitment of circulating leucocytes in the intestinal muscularis. Vagal afferents projecting to the CNS may be part of the cholinergic anti-inflammatory pathway which ensures that the response remains contained avoiding a detrimental extent of inflammation.