In previous experiments, we found that the threonine requirement of neonatal piglets fed parenterally was 40% of that when fed intragastrically; we hypothesized that much of the oral supply of threonine is being used for mucin production. To investigate this hypothesis, intragastrically fed 2-day-old piglets were fed one of three treatments for 8 days: 1) a threonine-adequate diet (IG-A; 0.6 g threonine.kg(-1).day(-1) fed intragastrically); 2) a threonine-deficient diet (IG-D; 0.1 g threonine.kg(-1).day(-1) fed intragastrically); or 3) a threonine-deficient diet with adequate threonine delivered parenterally (IV-A; 0.5 g threonine.kg(-1).day(-1) fed parenterally plus 0.1 g threonine.kg(-1).day(-1) fed intragastrically). IG-D piglets experienced higher nitrogen excretion, higher plasma urea, and lower plasma threonine concentrations versus both of the other groups (P < 0.05), indicating profound threonine deficiency. Mucosal mass and total crude mucin content were lower in the colons of IG-D pigs (P < 0.05). Histopathological analysis showed lower numbers of acidic mucin-producing goblet cells in the duodenum and ileum of IG-D pigs. In IG-D pigs, acidic mucin subtypes were lower in the small intestine but higher in the colon, which corresponded with persistent diarrhea. The parenteral supply of threonine was adequate to maintain most outcome parameters, although IV-A pigs did have smaller colonic goblet cells with more acidic mucins compared with IG-A pigs. Overall, our results suggest that adequate dietary threonine was critical in the production of mucus and that a parenteral threonine supply can ameliorate most of the symptoms of oral threonine deficiency.