Flaxseed hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on its value for dairy production. Monensin supplementation is known to modify biohydrogenation of FA by rumen microbes. Therefore, the main objective of the experiment was to determine the effect of feeding a combination of monensin and flaxseed hulls on ruminal fermentation characteristics and FA profile of ruminal fluid and milk. Four ruminally fistulated multiparous Holstein cows averaging 665±21 kg body weight and 190±5 d in milk were assigned to a 4×4 Latin square design (28-d experimental periods) with a 2×2 factorial arrangement of treatments. Treatments were: 1) control, neither flaxseed hulls nor monensin; 2) diet containing (dry matter basis) 19·8% flaxseed hulls; 3) diet with monensin (16 mg/kg dry matter); 4) diet containing 19·8% (dry matter basis) flaxseed hulls and 16 mg monensin/kg. Flaxseed hull supplementation decreased the acetate to propionate ratio in ruminal fluid and monensin had no effect. Concentrations of trans-18:1 isomers (trans9,trans11,trans13/14+6/8) and cis9,12,15-18:3 in ruminal fluid and milk fat were higher and those of cis9,12-18:2 in milk fat tended (P=0·07) to be higher for cows supplemented with flaxseed hulls than for cows fed no flaxseed hulls. Monensin had little effect on milk fatty acid profile. A combination of flaxseed hulls and monensin did not result in better milk fatty acid profile than when feeding only flaxseed hulls.