The use of such agricultural by-products as rice straw is important in fulfilling the needs of feed for ruminants, especially during the dry season. However, the main problem of using rice straw a feedstuff is low digestibility due to the existence of the links between lignin and cellulose or hemicellulose. This study was designed to evaluate the effects of the application of biostarter (BS) developed from rumen bacterial of buffalo on the of the changes of the chemical composition of the rice straw under a different duration of fermentation time. The experiment was carried out in a factorial arrangement according to a completely randomized design. The first factor was different levels of BS application, i.e. rice straw without BS (a1), rice straw + 5% BS (a2), rice straw +10% BS (a3), and rice straw + 15% BS (a4). The second factor was the duration of the fermentation time, i.e., two weeks (b1) and four weeks (b2). Each treatment combination was repeated three times, giving the total number of the experimental unit was 24. The results of the study indicated that increased levels of BS had a significant (P<0.05) effect on the concentration of crude protein, crude fiber, ash, and nitrogen-free extract (NFE) of the rice straw but no effects (P>0.05) on lipid content. The length of the fermentation period only affected (P<0.05) the crude fiber and crude protein content but no effects (P>0.05) on NFE, lipid, and ash. There was no interaction between levels of BS and the duration of the fermentation time on the chemical components of the rice straw. In conclusion, the use of BS was successfully changing the chemical composition of rice straw.