The purpose of the study was to investigate whether the reported increased cardiovascular disease risk in shift workers could be explained by changes in cardiovascular risk factors. In a cohort of 239 shift and 157 daytime workers, 1-year changes in biological and lifestyle cardiovascular risk factors were monitored between the start of a new job and 1 year later. Both body mass index and low-density lipoprotein/high-density lipoprotein cholesterol ratio decreased significantly in shift workers compared with daytime workers (body mass index change: -0.31 and +0.13 kg/m; low-density lipoprotein/high-density lipoprotein ratio change: -0.33 and -0.13 respectively). Cigarettes smoked per day increased significantly in shift compared with daytime workers (+1.42 and -1.03, respectively). Therefore, only for smoking, an unfavorable change was observed. This may explain, at most, only a part of the excess cardiovascular disease risk reported in shift workers.