Campylobacteriosis in humans is associated with handling and consumption of contaminated broiler meat. Reduction of the number of Campylobacter-colonized broiler flocks could potentially be realized by decreasing their susceptibility for colonization. The aim of this study was to determine the effect of feed supplementation with a mixture of medium chain fatty acids (C-8-C-12) on susceptibility of broilers for Campylobacter colonization, feed conversion and body weight gain. Two experiments were carried out with individually housed commercial broilers. The birds were fed with medium chain fatty acids supplemented feed (n = 227), or received feed without supplement (n = 87). The birds were inoculated with a dose of Campylobacter jejuni varying between log(10)1.19-5.47 CFU. During 14 days after inoculation, cecal or fecal samples were collected, in which the presence of C jejuni was determined by bacterial culture. Beta-binomial dose-response modeling of the colonization status at 14 days post-inoculation was performed to estimate the C. jejuni dose necessary to colonize 50% of inoculated broilers, which was estimated to be 200 times higher in broilers fed with supplemented feed (log(10)4.8 CFU) than in control broilers (log(10)2.5 CFU). Feed conversion was not affected by feed supplementation, while body weight gain was 49 g higher in broilers fed with supplemented feed. These findings indicate that susceptibility of broilers for Campylobacter colonization is decreased by supplementation with medium chain fatty acids, and that feed supplemented with this mixture may be a promising tool for the reduction of Campylobacter colonization in commercial broiler flocks.