Owing to vital roles played by enzymes in the preservation of the make-up of soil ecosystem and functional diversity, the influence of organic manure on the resultant biological quality of a crude oil polluted agricultural soil from a 90 d phytoremediation pot experiment was investigated. A 4-factor phyto-assisted clean-up of crude oil polluted agricultural soil was designed with options of manure amendments, to boost micobial activities. Profiles of β-glucosidases, proteases, dehydrogenases, phosphomonoesterases and respiration were investigated. Analysis of variance of triplicate experiment was carried out. Application of soil conditioner gave no marked dehydrogenase activity, which increased with depletion of available phosphorus. Marked increases in CO 2 release and alkaline phosphatase activity with soil conditioning may implicate beneficial relationship with the abundance of microbial populations. Flooding in some pots correlated with β-glucosidase and respiratory acitivities. A direct relationship between cellulose breakdown, measurable with β-glucosidase activity, organic matter and CO 2 release, measurable with respiratory activity within all soils was found in the present study. Use of organic manure significantly improved CO 2 release by soil biota in hydrocarbon-impacted soil and may be explored for phytoremediation technique.