Experiments were initiated to select a sterilization method(s) that minimizes alterations in the digestive properties of cereal grains and, thus, would be suitable for the study of cereal grain digestion by pure cultures of ruminal bacteria. The following five treatments were examined: unsterilized (U), autoclaving with buffer (AB), autoclaving without buffer (AD), ethylene oxide (E), and gamma irradiation (I). Solubility of DM, starch, and CP was determined by soaking grain in buffer for 1 h followed by filtration through Whatman #54 filter paper. Ground corn and wheat from each treatment were placed in vials with a 1:1 mixture of Bryant's medium and ruminal inoculum. Vials were incubated for 4, 8, 12, 24, and 48 h and analyzed for starch content. Bacterial growth was not evident in sterilized, uninoculated samples. The AD treatment decreased the disappearance of CP in wheat and corn, whereas AB caused an increase in the disappearance of DM, CP, and starch in wheat (P less than .001) compared with U. Rates of microbial starch digestion for corn were 1.3, 1.5, 3.3, 14.7, and 3.5%/h and for wheat were 1.3, 3.4, 4.6, 17.1, and 4.6%/h for AD, E, I, AB, and U, respectively. Contrasts indicated that AD and AB differed (P less than .001) from U for both corn and wheat. It is likely that gelatinization of cereal starch enhanced microbial starch digestion in AB and the formation of Maillard products reduced starch digestion in AD. Corn and wheat sterilized with E or I had digestive properties that closely resembled those of U grain, and either sterilization method was suitable for studying cereal grain digestion.