Exposure to Coxiella burnetii is a risk in the Hunter New England (HNE) region of New South Wales (NSW), Australia, based on yearly reported cases of Q fever. We assessed seroprevalence of phase II antibodies to C. burnetii by indirect immunofluorescence assay (IFA; screening at 1/50 dilution) of residents of 24 local government areas (LGA) of the HNE region of NSW. A total of 2,438 randomly selected sera sent to the Hunter Area Pathology Service for routine diagnostic purposes (not Q fever testing) during the period of 2006-2009 were tested. The overall seroprevalence in sample group was 7%. The proportion of males (59%) was higher than females (41%). In age distribution, the largest proportion (37%) of seropositives was in the > 60 years age group. Lower prevalence was observed in 0-9 years (1%) and 10-19 years (5%) age groups. The seroprevalence in different LGAs varied between 0.5% and 22%. It was highest in Guyra (22%), Gunnedah (21%), Tenterfield (18%), and Narrabri (16%), with Newcastle (0.5%), Port Stephens (2%), Lake Macquarie (3%), and Singleton (3%) being the lowest. In most of the LGAs, seroprevalence was between 6% and 12%. This report indicates a considerable exposure to C. burnetii of residents in rural areas of the HNE region and is consistent with the high notification rate for Q fever in this part of Australia.