Japanese encephalitis virus (JEV) transmission in temperate Australia has underscored a critical need to characterise transmission pathways and identify probable hosts of the virus. This systematic review consolidates existing research on the vertebrate hosts of JEV that are known to exist in Australia. Specifically, we aim to identify probable species involved in JEV transmission, their potential role as hosts and identify critical knowledge gaps. Data were extracted from studies involving experimental infection, seroprevalence and virus isolation and were available for 22 vertebrate species known to reside in Australia. A host competence score was calculated to assess the ability of each species to generate and sustain a viraemia. Based on the host competence score and ecology of each species, we find that ardeid birds, feral pigs and flying foxes have potential as maintenance hosts for JEV in the Australian context. We also note that domestic pigs are frequently infected during outbreaks, but their role as amplification hosts in Australia is unclear. Evidence to confirm these roles is sparse, emphasising the need for further targeted research. This review provides a foundation for future investigations into JEV transmission in Australia, advocating for enhanced surveillance and standardised research methodologies to better understand and mitigate the virus's impact.