Abstract. Background and Objective: Protection of proteins is essential for productive ruminants, this study aimed to obtain optimal gambier leaf residue (GLR) that can protect the protein of cattle feed supplement (CFS) from rumen microbial degradation. Materials and Methods: Gambier (Uncaria gambir Roxb.) leaf residue containing 9.96% condensed tannin was added to CFS containing 29% crude protein with a mixture of brown sugar, soybean meal, coconut cake, tapioca, urea, and minerals at level 0 (control), 2.5%, 5.0%, and 7.5%. Condensed tannin content in CFS: 0%, 0.68%, 1.17% and 1.60% then which were tested in rumen digestion in vitro. Results: Degradation of crude protein in the rumen decreased linearly with an increase in tannin level and ratio of tannin/ protein in CFS (R2 = 0.40; P <0.05) with degradation of 69.87% to 56.37%. Rumen undegraded dietary protein (RUDP), TCA-soluble N and peptide N was obtained respectively in the range (49.91-74.07; 319.14-465.20; 304.40-451.25) mg/ 100 ml rumen fluid, the lowest at control and highest at level 5.0% GLR. NH3-N concentration decreased linearly with an increase in GLR level while the rumen pH was neutral and not significant with GLR level. Conclusions: The optimum level of GLR in CFS was obtained 4.88% with the minimum rate of protein degradation 14.99 to 1.05%/ hour with the lowest rate at 3 hours and the highest at 48 hours of incubation.
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