The lipid distribution and fatty acid (FA) composition of total lipids, polar lipids and free fatty acids (FFA) were determined in liquid-associated bacteria (LAB) and solid-adherent bacteria (SAB) isolated from the rumen contents of seven dairy cows fitted with rumen fistulas. Two experiments, arranged according to a 4 x 4 and 3 x 3 Latin Square design, were performed using two basal diets consisting of one part hay and one part barley-based concentrate, and five lipid-supplemented diets consisting of the basal diet plus (g/kg dry matter):53 or 94 rapeseed oil, 98 tallow, 87 soya-bean oil or 94 palmitostearin. For all diets used, total lipids were 1.7-2.2 times higher in SAB than in LAB (P less than 0.05); this probably resulted from a preferential incorporation of dietary FA absorbed onto food particles. Addition of oil or fat to the diets did not modify the polar lipid content but increased the FFA content of SAB and LAB by 150%. Lipid droplets were observed in the cytoplasm in SAB and LAB using transmission electron microscopy, which suggested that part of the additional FFA was really incorporated into the intracellular FFA rather than associated with the cell envelope by physical adsorption. Linoleic acid was specifically incorporated into the FFA of SAB, which emphasized the specific role of this bacterial compartment in the protection of this acid against rumen biohydrogenation.