Infection is a major concern in intestinal transplant recipients. Bacterial migration to extraintestinal sites is a central component of the gut hypothesis of sepsis. However, some studies have cited the beneficial effects of bacterial translocation (BT) on the host acquired immune system. We evaluated the role of previous BT on a subsequent BT challenge, examined the BT index in organs as well as changes in white blood cell (WBC) count in mesenteric lymph and blood for correlation with outcomes. Wistar rats (n = 60) were divided into a BT group (n = 20), which underwent inoculation of 10 mL of 10(10) CFU/mL Escherichia coli R-6 confined to the small intestine as opposed to a BT1-14 group (n = 20), which underwent the BT procedure on days 1 and 14 or a S1-BT14 group (n = 20) that received 10 mL of saline on day 1 and the BT procedure on day 14. Half of the animals were killed 2 hours following the BT procedure. Samples from different compartments were collected for culture. Mesenteric lymph and peripheral blood were examined for WBC counts. The other half of the hosts was subjected to outcome evaluation concerning weight gain and mortality. Animals undergoing double BT showed a significantly lower index of bacterial recovery (liver, spleen, and blood) compared with those having a single BT (P < .05). The WBC count of mesenteric lymph cells after double BT was similar to naïve animals, but significantly lower than the single BT group (P < .05). The outcome was unchanged among double BT versus other groups. A previous BT challenge was efficient to generate a host-defense mechanism against a second BT episode induced by intestinal overgrowth with the same bacterial strain.