Introduction Disruption of the intestinal epithelial barrier has been shown to occur following burn injury. This process can lead to translocation of pathogens from the gut lumen to the systemic circulation and distant organs thereby increasing the risk for sepsis. The aim of this study was to examine the effect of amifostine (WR-2721) on bacterial translocation in a rat burn injury model. Materials and Methods A total of 27 male Wistar albino rats were divided into three groups of nine. Group I was a control group. Group II was subjected to third-degree burns over 30% of the total body surface area, and group III was administered amifostine 200 ml/kg intraperitoneally, followed by a 10 ml/kg/day maintenance dose after undergoing third-degree burns. After 48 hours, tissue and blood samples were obtained and cultured from the liver, spleen, mesenteric lymph nodes, and cecum. Results Blood cultures were negative in all groups. In the control group, colonization appeared only in the cecum, but in groups II and III, colonization was found in the liver, spleen, mesenteric lymph nodes, and cecum. While bacterial colonization was most frequently found in the cecum and mesenteric lymph nodes, bacterial counts did not significantly differ in the cecum (P = 0.298) and mesenteric lymph nodes (P = 0.418) between groups II and III. Conclusions Amifostine alone is not effective in controlling bacterial translocation associated with burn injuries. These results should be interpreted with caution as there are a number of factors that affect bacterial translocation.