To assess the levels of stress and the prevalence of cardiovascular disease (CVD) risk factors in female law enforcement officers (LEOs). Self-reported data including job-related stress and CVD risk factors were obtained from 65 female LEOs. Stress scores were compared with 429 males LEOs and CVD risk factors were compared with 1,213 demographically similar female respondents to the CDC BRFSS survey. All subjects resided in Iowa. Female LEOs had more stress (perceived stress, p < 0.01), more job-related stress (job strain, vital exhaustion and effort-reward imbalance, p < 0.01 for all), but similar social support (social provision scale, p = 0.412) than male LEOs. Female LEOs had a significantly higher prevalence of hypercholesterolemia than the general Iowa female population (46.2 vs. 29.3%, p < 0.01). There was a trend towards a higher prevalence of diabetes in female LEOs (9.2 vs. 4.5%, p = 0.084). The most commonly cited contributor to their perceived CVD risk was stress (77%). Female LEOs who felt that being either a LEO (67.7%) or a female LEO (41.5%) contributed to their risk for chronic diseases had more stress and a higher prevalence of overweight and obesity than female LEOs who felt differently. Female LEOs have higher stress than male LEOs. The prevalences of hypercholesterolemia and diabetes are higher in female LEOs than that of the general female population. Thus, female LEOs may be at a greater risk for CVD than their male counterparts.