To investigate the effects of silver nanoparticles on the histological structure and properties of the mucosubstances in the intestinal mucosa, Sprague-Dawley rats were divided into four groups (10 rats in each group): vehicle control, low-dose group (30 mg/kg), middle-dose group (300 mg/kg), and high-dose group (1,000 mg/kg), and administered silver nanoparticles (60 nm) for 28 days, following OECD test guideline 407 and using GLP. The control sections contained no silver nanoparticles; however, the treated samples showed luminal and surface particles and the tissue also contained silver nanoparticles. A dose-dependent increased accumulation of silver nanoparticles was observed in the lamina propria in both the small and large intestine, and also in the tip of the upper villi in the ileum and protruding surface of the fold in the colon. The silver nanoparticle-treated rats exhibited higher numbers of goblet cells that had released their mucus granules than the controls, resulting in more mucus materials in the crypt lumen and ileal lumen. Moreover, cell shedding at the tip of the villi was frequent. Lower amounts of neutral and acidic mucins were found in the goblet cells in the silver nanoparticle-treated rats, plus the amount of sialomucins was increased, while the amount of sulfomucins was decreased. In particular, in the colon of the silver nanoparticle-treated rats, sialyated mucins were detected in the lamina propria, the connective tissue under the epithelia. Therefore, the present results suggest that silver nanoparticles induce the discharge of mucus granules and an abnormal mucus composition in the goblet cells in the intestines.