Analysing the physicochemical characteristics of water and soil in conservation areas gives information about their quality and the effective management strategies wildlife conservationists could adopt. The study assessed thephysicochemical characteristics of selected perennial rivers and soil samples in Old Oyo National Park, Nigeria. A totalof 24 composited surface water and 36 topsoil samples were collected using grab and random sampling techniques,respectively for four seasons (two dry and two wet) for two consecutive years (2017 and 2018). The samples were analysed for selected physicochemical characteristics using standard methods. Data collected were subjected to descriptive and inferential (ANOVA) statistics SPSS (version 20.0) at α0.05. The result showed that the mean values (in the water samples) of Total Suspended Solids (673.13±592.10) in dry season 2018, Total Solids (799.37±610.17) in dry season 2018, and Sulphate (469.34±354.94) in dry season 2017 were above the comparable WHO permissible limit while the mean values (in the soil samples) of total nitrogen (except dry season 2018), exchangeable Magnesium and Potassium (across all the seasons) were above the comparable critical limits. There were significant differences in all the physicochemical characteristics of water sampled [except pH (P=0.12), chloride (P=0.96) and BOD (P=0.86)] while organic carbon (P=0.047), organic matter (P=0.041), nitrogen (P=0.020), calcium (P=0.016), total exchangeable bases (P=0.009) and effective cation exchange capacity (P=0.033) in soils were significantly different across the seasons of sampling. The soil physicochemical parameters above comparable critical limits may have elicited from the impact of anthropogenic activities by the surrounding communities with possible implication on wild animal health in the park.