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A significant portion of the burden of illness of workers in Canada and the United States is attributable to poor lifestyle habits, namely smoking, physical inactivity and low consumption of fruits and vegetables. The Healthy Enterprise Standard (HES) targets four intervention areas, including one to promote healthy lifestyle habits. The aim of this study was to evaluate the impact of HES implementation on workers’ smoking behaviour, physical activity, consumption of fruits and vegetables as well as their self-rated health status.This was an intervention study with a before-after design derived from secondary data. Organisations adopted the standard of their own initiative and were responsible for implementing interventions. All active employees of ten Quebec organisations were solicited to participate before (T1=2849) and 24–38 months (T2=2560) following the standard’s implementation. Smoking status, weekly frequency of physical activity and daily fruit and vegetable consumption were evaluated at T1 and T2 using self-report items elaborated from a national survey. Self-rated health status was assessed at both time points using one validated self-report item. Intervention exposure was measured by questionnaire and complemented by qualitative analyses.Between T1 and T2, the prevalence of weekly physical activity frequency increased for men and the prevalence of ex-smokers increased for both men and workers aged 55 or older. When body mass index was taken into account, the prevalence of daily fruit and vegetable consumption also increased for both men and workers aged 55 or older. However, these workers’ self-reported health status remained unchanged from T1 to T2. No clear changes were observed in other age groups.The improvement of lifestyle habits for both male workers and those aged 55 or older following HES interventions is significant since scientific literature suggests that those groups are generally less inclined to participate in health promotion workplace interventions.