The adherence to the Mediterranean diet is beneficial for cardiovascular diseases prevention. The study aim is to use Mediterranean lifestyle (MEDLIFE) questionnaire for estimation of Mediterranean lifestyle habits among the working population and to establish MEDLIFE score correlation with the risk factors for cardiovascular diseases. In the study has participated 366 workers from Croatia, which fulfilled MEDLIFE and validated food frequency questionnaire (FFQ) questionnaire. The multivariate logistic regression was performed to evaluate the association between MEDLIFE index, workers’ obesity and cardiovascular diseases risk. The lowest adherence to Mediterranean lifestyle was associated to younger, low education, body fat above acceptable ranges and unfavorable lipid profile. Significant association to Mediterranean lifestyle was more among women (p = 0.002), middle aged (p = 0.02), highly physically active (p = 0.009) and those who play collective sports >2 h/w (p = 0.001), having body fat within acceptable range (p = 0.003), total cholesterol less (p = 0.03) and high-density lipoproteins (HDL-C) (p = 0.04) more than recommended. Inverse significant association was for high educational level (p = 0.02). The Mediterranean lifestyle adherence is associated to lower risk factors for cardiovascular diseases among studied working population. MEDLIFE index revealed that physical activity and conviviality are better ingrained among younger population but not the Mediterranean diet.