A cross-sectional survey of staff of the New South Wales Department of Agriculture for prior exposure to Q fever was conducted using the complement fixation test, indirect immunofluorescent antibody test, a delayed hypersensitivity skin test, a standard questionnaire, and a supplemental history, with the aim being to determine the proportion of employees that have been in contact with Q fever and the jobs that pose the greatest risk of exposure to the disease. Of 829 employees, 89 (10.7%) tested positive, with those handling livestock being more likely to have been exposed to Q fever than employees in low-risk occupations. This difference reached statistical significance (P < 0.01) when employees with other risk factors for exposure to Q fever were excluded. Veterinarians, stock inspectors, and regulatory officers had the highest risk of previous exposure. This study confirms that Q fever is a disease related to occupations that involve handling livestock, and it provides a basis upon which to promote vaccination of agricultural workers.