The study aimed to describe the prevalence of amblyopia and associated refractive errors among an adult Australian population. The Visual Impairment Project (VIP) is a population-based study of age-related eye disease in the state of Victoria, Australia. Data were collected through standardised interviews and orthoptic and ophthalmic dilated examinations. Amblyopia was defined as best-corrected visual acuity of 6/9 or worse in the absence of any pathological cause. The participants were 3,265 urban residents and 1,456 rural residents of the VIP ranging in age from 40-92 years (mean = 59 years; 53% female). The prevalence of unilateral amblyopia was 3.06% (95% C.I. 2.59, 3.53). Amblyopia was not found to be statistically different by age group (p=0.096), gender (p=0.675), or place of birth (p=0.14). Anisometropia was statistically more common (p<0.001) in amblyopic cases (51.1%) compared to the normal population (9.7%), and 54% of amblyopic eyes had visual acuity of worse than 6/12. Amblyopia is a significant cause of unilateral reduced visual acuity in a population aged 40 years and older. Anisometropia was more prevalent and the degree of anisometropia was greater in the amblyopic group compared with the normal population. Oblique astigmatism was more prevalent in the amblyopic group compared with the normal population.