The effect of zinc deficiency on intestinal absorption of cholesterol was investigated in adult male rats fed 3.6 ppm of dietary zinc (ZD) and compared with pair-fed (PF) and ad-libitum (CT) controls fed 30.6 ppm of zinc for 10 to 12 wk. The intestinal absorption of cholesterol was measured by collecting the mesenteric lymph at hourly intervals for 8 h after infusing a dose of [14C]cholesterol in a lipid emulsion through an indwelling duodenal catheter. Data on the hourly lymphatic output of [14C]cholesterol showed that the absorption of [14C]cholesterol plateaued at 4 h postdosing in all groups. The rate of the appearance of [14C]cholesterol in the mesenteric lymph significantly decreased in ZD rats at 5 h and thereafter. The cumulative absorption of [14C]cholesterol by ZD rats for the 8-h period was 4.4 +/- 1.0% of the dose, compared with 11.8 +/- 1.1% by PF and 10.7 +/- 1.4% by CT rats. There were no differences in percent distribution of esterified [14C]cholesterol among the three groups. Also, no significant change in mucosal cholesterol esterase activity was observed due to zinc deficiency. The present data and previous findings taken together suggest that the primary mechanism responsible for the impaired absorption of cholesterol may be associated with defective formation of chylomicrons in the intestinal mucosa.