Effective groundwater management in semi-arid regions requires detailed aquifer characterization, particularly in ecosystems facing ecohydrological stress. This study characterizes aquifer systems in the southern Kruger National Park (KNP) to support the conceptualization and protection of groundwater-dependent ecosystems (GDEs). Pre-existing single-well and multiple well constant rate pumping test data from five boreholes were analyzed to estimate key aquifer parameters. Borehole logs and geological cross-sections were used to identify preferential groundwater flow paths and delineate potential recharge and discharge zones. Groundwater recharge and evapotranspiration were estimated using the Cumulative Rainfall Departure (CRD) method, the Rainfall Infiltration Breakthrough (RIB) method, and remotely sensed FAO datasets. Results show that the region's aquifers consist of shallow weathered and deeper fractured granitic/gneiss units of the Archaean Nelspruit formation, with thicknesses ranging from 299 to 383 m. Transmissivity values ranged from 0.5 to 11.2 m2/day and Storativity from 1.28 x 10-6 to 4.38 x 10-4, indicating generally low aquifer permeability except in localized fractured zones. Dyke intrusions were found to influence groundwater occurrence and the distribution of GDEs. The study underscores the importance of integrated hydrogeological analysis for accurately conceptualizing GDEs beyond surface indicators that overlook subsurface connectivity. It provides a foundational understanding of aquifer-ecosystem interactions and highlights the need to incorporate these relationships, along with spatial and temporal groundwater variability, into numerical models. This approach supports more effective simulation, protection, and policy development for GDE management.