The waterways or ephemeral systems of the South Australian Murray Darling Basin (SA MDB) rangelands are unique to the area and have not been previously studied to create a comprehensive baseline for flora and fauna. This lack of information greatly increases the risk of poor decision making and limits the decisions regarding management strategies and their impacts within the rangelands. The SA MDB rangelands biological survey has drawn on existing information and newly surveyed information together to provide natural resource managers with a more comprehensive identification of the unique assets of the rangelands. The biological survey focused on collecting baseline biological data including aquatic and terrestrial flora and fauna at eight permanent water sites. Eight formal Department for Environment and Heritage Biological Survey sites were established, where data was collected on vegetation, terrestrial mammals, reptiles, amphibians, and birds. All fauna was also opportunistically sampled to distance of 1 km from each survey site.