The physico-chemical and microbial examination of Ajali River in Ezeagu Local Government Area (LGA) of Enugu State, South-eastern Nigeria, were carried out to assess the characteristics of the water for potability. Water samples were collected from three sampling points, including the one close to the brewery effluent discharge into the river and two different points that were affected by non-point sources. Altogether, eighteen samples were collected for two years during the dry, early and late rainy seasons. The physico-chemical, as well as microbial analyses of the samples, were determined following the standard methods. Significant spatial and seasonal variations (p<0.05) were clear for some physico-chemical parameters such as pH, phosphate, and sodium. Mean values of the Water Quality Index (WQI) were: dry season (104.23±9.75); early rainy season (113.22±8.78) and the late rainy season (102.58 ± 11.15). All the values are above 100 showing that the calculated WQI in all the seasons is not good however after the addition of lime, boiling and filtering to increase the quality.