The effects of wet distillers grains with solubles (WDG) on in vitro rate of gas production, IVDMD, H(2)S production, and VFA were evaluated. Five substrate treatments that were balanced for ether extract content were arranged in a 2 x 2 + 1 factorial. Factors were concentration (15 or 30%; DM basis) and source of WDG (corn or sorghum WDG; CDG and SDG, respectively) plus a 0% WDG control in substrates with steam-flaked corn as the basal grain. Control substrates had greater (P < 0.01) IVDMD and total gas production per gram of substrate DM than WDG-based substrates, and IVDMD was greater (P = 0.03) for CDG than for SDG substrates. Increasing WDG inclusion from 15 to 30% decreased IVDMD and total gas production (P < 0.05), but H(2)S production (micromol/g of fermentable DM) increased (P = 0.01) as inclusion of WDG increased. There were no differences (P > or = 0.10) among treatments in proportions of major VFA, acetate:propionate ratio, and total VFA concentration. These results suggest that including WDG in the substrate decreased IVDMD and gas production, which was particularly evident as WDG increased from 15 to 30% of substrate DM. In addition, CDG seemed to be more digestible than SDG. Hydrogen sulfide production increased with increasing WDG in the substrate, reflecting greater S concentrations in WDG, but in vitro VFA profiles were not affected by WDG concentration or source.