This study was conducted to evaluate the effects of dietary arginine levels on microvascular development of the small intestine in early-weaned pigs. Twenty-four crossbred pigs (5.0 +/- 0.3 kg body weight) were individually housed and randomly allotted to 1 of 3 diets supplemented with 0, 0.7, and 1.2% L-arginine (8 pigs per group). Pigs consumed the diets ad libitum for 10 d. We collected blood samples on d 3, 6, and 10. On d 10, 6 pigs from each group were randomly selected and killed for tissue sample collection. Compared with control pigs, dietary supplementation with 0.7% L-arginine increased (P < 0.05) jejunal concentrations of nitrite and nitrate (stable oxidation products of nitric oxide), intestinal villus height, as well as plasma proline and arginine concentrations on d 6 and 10. Dietary supplementation with 0.7% L-arginine also increased (P < 0.05) immunoreactive expression of CD34 in duodenal submucosa, ileal mucosa and submucosa, and expression of vascular endothelial growth factor (VEGF) in duodenal submucosa, jejunal mucosa and submucosa, and ileal mucosa compared with the control and 1.2% L-arginine supplementation. Dietary supplementation with 1.2% L-arginine increased (P < 0.05) the concentration of jejunal endothelin-1 compared with the control pigs. Immunoexpression of VEGF in duodenal mucosa and plasma lysine concentrations on d 6 and 10 were lower (P < 0.05) in pigs supplemented with 1.2% L-arginine than in unsupplemented pigs. Collectively, these findings indicate that the effects of L-arginine on microvascular development are beneficial at lower levels but have adverse effects at higher intakes. Dietary supplementation with 0.7% L-arginine may be a useful method to improve microvascular development in the small intestine of early-weaned pigs.