Background: Biliary secretion is generally considered to be an obligate step in the pathway of excess cholesterol excretion from the body. We have recently shown that an alternative route exists. Direct transintestinal cholesterol efflux (TICE) contributes significantly to cholesterol removal in mice. Aim: To investigate whether the activity of this novel pathway can be influenced by dietary factors. In addition, we studied the role of cholesterol acceptors at the luminal side of the enterocyte. Methods: Mice were fed Western-type diet (0.25 % wt/wt cholesterol; 16% wt/wt fat) a high fat diet (no cholesterol; 24% wt/wt fat) or high cholesterol diet (2% wt/wt) and TICE was measured by isolated intestinal perfusion. Bile salt/phospholipids mixtures served as cholesterol acceptor. Results: Western type and high fat diet increased TICE by 50% and 100 %, respectively. In contrast, the high cholesterol diet did not influence TICE. Intestinal Scavenger Receptor class B Type 1 (Sr-B1) mRNA and protein levels correlated with the rate of TICE. Unexpectedly, although confirming a role for Sr-B1, TICE was significantly increased in Sr-B1 deficient mice. Apart form the long term effect of diets on TICE, acute effects by luminal cholesterol acceptors were also investigated. The phospholipid content of perfusate was the most important regulator of TICE, bile salt concentration or hydrophobicity of bile salts had little effect. In conclusion: TICE can be manipulated by dietary intervention. Specific dietary modifications might provide means to stimulate TICE and, thereby to enhance total cholesterol turnover.