Hendra virus causes acute and highly fatal infection in horses and humans. Pteropid bats (flying-foxes) are the natural host of the virus, with age and species being risk factors for infection. Urine is the primary route of excretion in flying-foxes, with viral RNA more frequently detected in Pteropus alecto and P. conspicillatus than other species. Infection prevalence in flying-foxes can vary between and within years, with a winter peak of excretion occurring in some regions. Vertical transmission and recrudescing infection has been reported in flying-foxes, but horizontal transmission is evidently the primary mode of transmission. The most parsimonious mode of flying-fox to horse transmission is equine contact (oro-nasal, conjunctival) with infected flying-fox urine, either directly, or via urine-contaminated pasture or surfaces. Horse to horse transmission is inefficient, requiring direct contact with infected body fluids. Flying-fox to human transmission has not been recorded; all human cases have been associated with close and direct contact with infected horses. Canine cases (subclinical) have also been limited to equine case properties. Notwithstanding the recent availability of an effective vaccine for horses, a comprehensive understanding of Hendra virus ecology and transmission is essential to limit inter-species transmission.