The kinetics of gas production during ruminal fermentation may provide valuable information about foods that can be used to formulate diets and model animal responses. However, measurement of digestion kinetics is affected by methodology and techniques must be established that provide accurate and precise estimates of kinetic parameters. Because gas production measurements provide the opportunity to estimate the digestion kinetics of both soluble and insoluble matter in foods, it would be desirable to use this technique on a wide variety of forages, grains, supplements, and by-product foods. Applying an in vitro technique to such a wide variety of substrates raises questions about the type of inoculum that should be used. The objective of our study was to evaluate the effects of donor animal and its diet on the measurement of gas production kinetics using both forage and concentrate substrates.