This study was conducted to investigate changes in fatty acid profiles of beef cows fed grass hay or barley silage based diets, with or without flaxseed supplementation. Both flaxseed and hay feeding increased levels of α-linolenic acid (LNA; 18:3n-3) in longissimus thoracis and backfat (P<0.001). A forage type by flaxseed level interaction was observed for most LNA biohydrogenation intermediates (P<0.05) that indicated feeding hay combined with flaxseed led to the greatest levels of total conjugated linolenic acid, total conjugated linoleic acid, total non-conjugated dienes and total trans-18:1. Predominant biohydrogenation intermediates included t11,c15-18:2, rumenic acid (c9,t11-18:2) and vaccenic acid (t11-18:1). Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.