Objectives: Nutrients passing the ileum induce mechanisms regulating pancreatic secretion, but the effect of short-chain fatty acids (SCFAs) present in the ileum because of either intestinal fermentation or due to the cecoileal reflux is still unclear. This study investigated the effect of ileal SCFAs on pancreatic secretion and plasma levels of peptide YY, cholecystokinin, motilin, and neurotensin. Methods: The pigs were fitted with pancreatic duct, ileal, and jugular vein catheters, and a duodenal T-shaped cannula. Saline, 85.0 or 170.0 mM acetate, 5.0 or 10.0 mM butyrate, 7.5 or 15.0 mM propionate were infused into the ileum during feeding. Results: The ileal infusions of SCFAs did not affect the pancreatic juice outflow and the lipase output. The protein output was lower when 10.0 mM butyrate or 170.0 mM acetate were infused. The trypsin output decreased for most of the SCFA infusions. The alpha-amylase output decreased for the infusion of 10.0 mM butyrate and tended to decrease for 170.0 mM acetate. The infusions did not change gut hormone level. Conclusions: Ileal SCFAs might induce an inhibition of pancreatic enzyme secretion under prandial conditions. Ileal SCFAs do not inhibit pancreatic secretion by a hormonal pathway involving the release of peptide YY, motilin, neurotensin, or cholecystokinin.