Our objective was to examine the effects of fructan supplementation on the immune response of weanling puppies subjected to bacterial challenge. Previous studies in bacterial challenged neonatal piglets have reported benefits of fructan supplementation. Thirty hound-cross puppies (12 wk of age) were used in a 2 x 3 factorial randomized complete block design. Following a 7-d baseline period, puppies were assigned to diets containing: 1) no prebiotic, 2) 1% short-chain fructooligosaccharides (scFOS), or 3) 1% inulin. After 14 d on treatment diet, dogs received an oral gavage of: 1) Salmonella typhimurium DT104 (5 x 10(8) colony forming units) or 2) 0.9% saline. Food intake, fecal and activity scores, body temperature, body weight, blood chemistry, intestinal nutrient transport, intestinal morphology and pathology, and gut microbiota were measured. Food intake decreased (P < 0.01) and body temperature increased (P < 0.05) in infected puppies. However, the decrease in food intake was less (P < 0.05) in those consuming fructans. Infected puppies consuming fructans also had decreased (P = 0.05) severity of enterocyte sloughing than those fed the control diet. Ileal Na+-dependent glucose transport was decreased (P = 0.02) in infected vs. noninfected puppies consuming CON, whereas no changes occurred in fructan-supplemented animals. Puppies consuming inulin also had increased fecal acetate (P = 0.03) and total short-chain fatty acid (P = 0.06) concentrations than scFOS-fed puppies and controls. Finally, puppies fed inulin had an increase (P = 0.05) in Lactobacillus concentrations compared with scFOS and CON. In summary, fructan supplementation appeared to attenuate some of the negative responses associated with Salmonella challenge and may provide protection against infection in weanling puppies.