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To prospectively study the association of night and shift work with weight change and lifestyle behaviors.Workers participating in the Netherlands Working Conditions Cohort Study (2008 and 2009) (N = 5951) reported night and shift work, weight and height. Groups included stable night or shift work, from day work to night or shift work, from night or shift work to day work, and no night or shift work in 2008 and 2009. Regression analyses were used to study association changes in night and shift work with weight change and changes in lifestyle behaviors.A larger weight change was seen in normal-weight workers changing from day to shift work (β = 0.93%; 95% confidence interval, 0.01 to 1.85) compared with stable no shift workers. No further associations of night and shift work with weight change were observed, neither in normal-weight, overweight, and obese workers.Despite the fact that starting night or shift work is associated with some unhealthy lifestyle habits, this study did not confirm a positive association of night and shift work with weight change over 1 year, except for normal-weight workers moving from day to shift work.