Fumonisin B₁ (FB1) alters intestinal epithelial cell cycle, absorptive, secretory and barrier properties of intestinal epithelial cells in vitro but data with intestinal mucosa are lacking. Therefore, we tested the hypothesis that acute exposure to FB1 alters indices of intestinal absorptive and secretory physiology and barrier function ex vivo. Eighteen non-littermates pigs (n=18) weaned at 28 d were slaughtered 7 d post-weaning. Jejunal tissue was collected immediately, the mucosa was mounted in Ussing chambers (UC) and short-term (2 h) effects of pure FB1 were investigated. Increased transepithelial resistance (TER) (P=0.05) and macromolecular passage (P=0.06) were observed after 60-90 min of incubation of jejunal mucosa with FB1. FB1 did not alter the jejunal absorptive and secretory physiology. In conclusion, 2h-exposure to FB1 tended to increase the trans- cellular permeability and increased para-cellular permeability of pig small intestine.