The global use of petroleum hydrocarbon for domestic and industrial purposes has necessitated the occurrence of the contamination of the living and non-living components of the environment. This study investigated the influence of water insoluble fractions of crude oil on physicochemical properties and microbes of soil obtained from a tertiary institution in Benin City, Nigeria using standard procedures. The result for soil physicochemical properties showed a significant increase (p ≤ 0.05) in total hydrocarbon, nitrate, phosphate and sodium ion, however nitrate ion decreased in day 28 and 42. Soil pH, electrical conductivity, calcium, potassium and magnesium ion concentration significantly decreased, although with very minute variations. Soil bacteria and fungi count also decreased significantly (p ≤ 0.05), with some variation in fungi count (p ≤ 0.05). The result suggest that treatment with water insoluble fraction of crude oil induced changes in the soil physicochemical and microbial component, which infers alterations in soil structure and component. This could negatively affect plant cultivation and growth, as well as animals and human that consume contaminated plants. Oil contamination should therefore be avoided or properly managed to prevent its detrimental effect on the non-ling and living soil component.