The bioavailability of selenium (Se) in high-Se yeast (SeY) was evaluated by measuring tissue Se accumulation and glutathione peroxidase (GSHPx) activity. For 4 weeks, 4-week-old male Wistar rats were fed a Torula yeast-based Se-deficient diet (basal diet) or a diet supplemented with a graded level (0.04, 0.08, 0.16, and 0.32 microgram/g) of Se as either sodium selenite or SeY, which was obtained from two different sources. Se supplementation did not influence growth, hematological values, or serum biochemical tests. Se contents and GSHPx activities in the liver, serum, and erythrocytes increased gradually with increases of the supplemented Se. At lower Se levels (0.04 and 0.08 microgram/g), selenite produced higher Se deposition and higher GSHPx activities than SeY did, but at a higher Se level (0.32 microgram/g), SeY showed higher measures. Strong correlations were detected between the supplementary Se levels and the tissue Se contents or GSHPX activities when the regression was fitted to this equation: R-Rb = m log X + k, where R represented tissue Se content or GSHPx activity in rats fed the diet supplemented with Se at X level, Rb corresponding mean value in rats fed the basal diet, m slope, and k constant. The bioavailability of Se in SeY, as assessed by slope ratio analysis using selenite as a reference Se, was 135% to 165% in the tissue Se content and 105% to 197% in the GSHPx activities. These results indicate that Se in SeY is more bioavailable than selenite Se, and therefore it is the preferred form for supplementation.