Fumonisin B(1) (FB1) alters intestinal epithelial cell cycle and absorptive, secretory, and barrier properties in vitro, but in vivo data are lacking. Therefore, we tested the hypothesis that repeated intake of a corn culture extract rich in fumonisins, mainly in FB1, alters indices of intestinal absorptive and secretory physiology and barrier function in vivo. Intra-litter pairs of pigs (n = 36) weaned at 28 d, were fed the vehicle (control) or the extract (providing 1.5 mg FB1/kg body weight) daily for 9 d starting 7 d postweaning. After slaughter, the jejunal mucosa of pigs was mounted in Ussing chambers (UC). Extract consumption for 9 d decreased the gain:feed ratio (P = 0.04) and increased liver weight (P = 0.01). Basal net ion secretion (P = 0.02), sodium-dependent glucose absorption (P = 0.02), and theophylline-induced secretion (P < 0.01) of the jejunal mucosa determined in UC were higher in pigs fed the extract than in controls. By contrast, jejunal permeability to the horseradish peroxidase model protein in UC was not influenced by extract consumption. Ileal villi tended to be longer (P = 0.07) and jejunal aminopeptidase N activity was lower (P < 0.01) in pigs fed the extract. In conclusion, consumption of an extract rich in fumonisins for 9 d has the potential to alter intestinal physiology, villous architecture, and enzyme activities. Underlying mechanisms remain to be investigated.