Diet digestibility and outputs of biohydrogenation intermediates were assessed in a continuous culture of ruminal microorganisms. Orchardgrass or red clover harvested and frozen during spring or fall served as the primary substrates for fermentation. During 10-d incubations, fermenters were fed thawed forage (50 g of DM/d), forage (42 g/d) plus 8 g/d of corn, or forage (34 g/d) plus 16 g/d of corn. Effluents from the last 3 d of incubation were composited for analyses. Starch input increased from 5 to 27% of DM as corn input increased from 0 to 16 g/d. Corn input reduced (P < 0.01) pH, increased (P < 0.01) microbial DM yield, and increased (P = 0.01) digestibility of DM, NDF, CP, and nonstructural carbohydrates. Overall, apparent hydrogenation (percentage) of cis9-18:1, 18:2n-6, and 18:3n-3 was greater (P < 0.05) with orchardgrass than clover. Hydrogenation of cis9-18:1 and 18:2n-6 increased (P = 0.01), but hydrogenation of 18:3n-3 decreased (P = 0.01) linearly due to corn input, regardless of forage. As a result, output of trans11, cis15-18:2 also decreased (P = 0.01). Average output of cis9,trans11-18:2 was greater (P = 0.01) for clover (1.3 mg/d) compared with orchardgrass (0.6 mg/d), but corn input with either forage increased (P = 0.01) cis9,trans11-18:2 output by 205%. Output of trans11-18:1 was greater (P = 0.01) from orchardgrass compared with clover (174 vs. 90 mg/d), but corn increased (P = 0.01) trans11-18:1 output only from clover fermentations. Output of trans10-18:1 was greater (P = 0.01) in response to orchardgrass compared with clover (10 vs. 4 mg/d), but corn addition doubled the output regardless of forage type. Output of trans10,cis12-18:2, which did not differ due to forage type, increased (P = 0.01) twofold in response to corn. Cis9,cis11-18:2 was a primary conjugated isomer produced from forage fermentations, but its output decreased (P = 0.03) in response to corn input. When inputs of 18:2n-6 plus 18:3n-3 were less than 0.9% of total DM (clover), hydrogenation was low (87%). When 18:2n-6 plus 18:3n-3 inputs were from 1.2 to 1.5% of total DM (orchardgrass), hydrogenation averaged 96%. Despite greater hydrogenation, incremental additions of cis9-18:1 and 18:2n-6 from corn grain increased (P < 0.05) outputs of trans10-18:1, trans11-18:1, trans10,cis12-18:2, cis9,trans11-18:2, and trans,trans-18:2 in effluent. Results suggest that forage species alone or in combination with corn grain can alter hydrogenation and profiles of intermediates to varying degrees.