The effect of three subtropical fiber-rich brown seaweeds, Sargassum hemiphyllum (Turn.) C. Ag., Sargassum henslowianum C. Ag. and Sargassum patens Ag., on protein bioavailability of casein were evaluated by growing rats. The effect of the fiber-rich seaweeds was determined by comparing the net protein ratio (NPR), true protein digestibility (TD), nitrogen balance (NB), biological value (BV), net protein utilization (NPU), and fecal and urinary nitrogen loss of seaweed-based diet groups with those of the cellulose-based control group. There were no significant differences on NPR and urinary N loss in all diet groups. However, the values of TD (range 85.6-89.6%), BV (range 96.9-97.5%), NPU (range 83.5-86.8%) and NB (range 145-150 mg/rat/day) in all seaweed-based diet groups were significantly (p<0.05, one-way ANOVA, Tukey-HSD) lower than those of the control group. On the contrary, the fecal weight (range 5.59-6.30 g/rat/day) and fecal nitrogen (N) loss (range 31.1-41.2 mg/rat/day) of rats fed seaweed-based diets were significantly (p<0.05, one-way ANOVA, Tukey-HSD) higher than that of the control one. However, the differences on the body weight gain and weight of internal organs including liver, kidney and spleen between the seaweed-based diets and control groups were insignificant except that the weight of caecum of rats fed seaweed-based diets was significantly (p<0.05, one-way ANOVA, Tukey-HSD) higher than that of control group. The possible mechanism of fiber-rich brown seaweeds in affecting the protein bioavailability of casein and their related nutritional implications were discussed.