Hepatitis E virus (HEV) is the only virus classified in the genus Hepevirus. This virus is emerging as a major public health concern in industrialized countries, where infection is believed to be zoonotic, through ingestion of undercooked contaminated pork or contact with infected animals. Even though this virus causes disease in a small proportion of infected individuals, more severe infection is possible and chronic infection has occurred in solid-organ transplant recipients and in patients with immunosuppressive disorders. In Australia, HEV is nationally notifiable, with the majority of cases reported in travellers. To date, there has only been one published locally acquired case, despite a HEV IgG seroprevalence up to 95% in commercial piggeries. In this study we measured HEV seroprevalence in a cohort of blood donors from around Australia. Plasma samples (n=3,237) were tested for HEV IgG (Wantai HEV-IgG ELISA, Beijing Wantai Biological Pharmacy Enterprise Co., Ltd.). Donor demographics, including age, gender, state of residence and overseas travel disclosure, were obtained. HEV IgG was detected in 6% (95% CI 5.18-6.81) of donors. As expected, seropositivity increased with increasing age (p<0.05). There was no difference in HEV IgG seropositivity with gender (p>0.05), nor between donors residing in the different states and territories (p>0.05). HEV IgG was detected in 3.37% (95% CI 1.63-5.10%) of donors not reporting overseas travel. Of these donors, 57% were males and 64% were >45 years of age. This study showed that for this sample population, 6% of Australian donors had serological evidence of a prior infection with HEV, which is similar to other non-endemic countries. However, we also demonstrated HEV seropositivity in donors who did not report international travel, suggesting locally acquired infection. Study of genotypes associated with locally acquired infection, to determine whether infection is from zoonotic or human-to-human transmission, would be of interest.