Platypus (Ornithorhynchus anatinus) populations are declining due to accelerating threatening processes including changing land use, river regulation, and climate change. With limited overland dispersal, the species is vulnerable to local extinctions that cumulatively impact overall population stability. Therefore, translocations may become an increasingly important tool for platypus conservation. A translocation of ten platypuses is scheduled for May 2023, with the objective of reintroducing platypuses to Sydney’s Royal National Park. No formal disease risk analysis (DRA) had previously been performed for monotremes, and challenges were encountered in the DRA process. Firstly, despite their iconic status, our understanding of platypus health and disease is constrained by the practical challenges associated with researching a semi-nocturnal, semi-fossorial, and semi-aquatic species. These knowledge gaps were important to explore during the DRA process and were a key consideration when deciding to use animals from single or multiple source populations. Fortunately, fieldwork associated with the translocation has presented a unique opportunity for pathogen-focused sample collection. This facilitated a more nuanced understanding of pathogen variation between different platypus populations and highlighted the value of multi-disciplinary translocation teams to maximise outputs of labour- and resource-intensive fieldwork. Secondly, given unique, species-specific parasites have evolved alongside the evolutionarily distinct platypus, it was important to formally capture the intrinsic value of conserving these species alongside their more charismatic host. Overall, the DRA process highlighted the importance of using first principles and avoiding over-simplifying disease ecology in the absence of robust and nuanced data.