Arginine plays an important role regulating nutrient metabolism, but the underlying mechanisms are largely unknown. This study was conducted to determine the effect of dietary arginine supplementation on the metabolome in serum of growing pigs using (1)H nuclear magnetic resonance spectroscopy. Sixteen 120-day-old pigs (48 +/- 1 kg) were randomly assigned to one of two groups, representing supplementation with 0 or 1.0% L: -arginine to corn- and soybean meal-based diets. Serum was collected after a 46-day period of treatment. Dietary arginine supplementation decreased fat deposition and increased protein accretion in the body. Principal component analysis showed that serum concentrations of low density lipoprotein, very low density lipoprotein, and urea were lower, but concentrations of creatinine, tricarboxylic acid cycle metabolites, ornithine, lysine and tyrosine were greater in arginine-supplemented than in control pigs. Additionally, the arginine treatment affected serum concentrations of nitrogenous and lipid signaling molecules (glycerophosphorylcholine and myo-inositol) and intestinal bacterial metabolites (formate, ethanol, methylamine, dimethylamine, acetate, and propionate). These novel findings suggest that dietary arginine supplementation alters the catabolism of fat and amino acids in the whole body, enhances protein synthesis in skeletal muscle, and modulates intestinal microbial metabolism in growing pigs.