It has been well established that a high level of stress is associated with medical problems, mental health difficulties and absenteeism at the workplace. The aim of this single-arm study design was to examine the potential effectiveness of a 12-session multidisciplinary stress reduction programme on reducing perceived stress and improving health behaviours and quality of life. One hundred and four women participated in a programme that incorporated group support, skill building and cognitive behavioural and relaxation techniques. A series of Bonferroni corrected t-tests found that the participants reported having significantly (p < 0.001) lower levels of perceived stress, improved health behaviours (sleep, nutrition, physical activity) improved overall health and improved quality of life at the end of the 12 week programme and at 1-month follow-up. Although the effect sizes for improvement were all large, there was no control group, so regression to the mean or selection bias may have impacted the results. Therefore, these results provide initial support for the implementation of gender-based worksite stress reduction programmes and provide guidance in designing an effective worksite stress reduction programme. Further research using randomized controlled trials is warranted.