The U.S. Air Force relies upon the C-17 Globemaster aircraft and aircrew for the effective and strategic transportation of materials, supplies, and manpower to sustain a diverse number of missions. This study utilized a web-based occupational health screening survey to examine the main sources of occupational stress and levels of psychological distress among active duty C-17 Globemaster pilots and loadmasters based in the continental United States engaging in short- and long-duration overseas flights in support of operational missions. Participants included 233 (67.93%) pilots and 110 (32.07%) loadmasters. Based on the number of aircrew assigned to the units surveyed, the response rate was 32%. Respondents answered questions regarding demographics, occupational factors, and sources of occupational stress. Levels of psychological distress were assessed using the Outcome Questionnaire 45.2. Qualitative analyses of textual responses to the items assessing self-reported sources of stress were performed. Descriptive statistics were calculated for the overall and subscale total scores. Univariate analyses of covariance and chi-square analyses were conducted to identify group differences between pilots and loadmasters on means and elevated levels of psychological distress, as well as subscales assessing symptom distress, interpersonal relations distress, and social role distress. Results indicated that the top sources of stress for both pilots and loadmasters were operational in nature (i.e., operational and administrative workload, organizational management issues, professional career development, and balancing work and home life demands and obligations). Aeromedical implications of the study’s findings and recommendations are discussed. Separate, yet cohesive, recommendations are provided for line and medical leadership to address the elevated levels of distress found among C-17 pilots. The recommendations are based on early identification and intervention of aircrew experiencing emotional and relational distress that may negatively impact performance. The value of routine, anonymous, organizational health stress screenings is also discussed along with limitations of assessments based upon genuine self-disclosure.