Understanding the effects of oil contamination on the composition and function of soil microbiota entails investigation of the effects of a mixture of hydrocarbons at the community level in a complex environmental matrix. One approach to this difficult problem is to ally a community-level fingerprinting approach with bioassays that have a physiological or functional implication. Two contrasting refined oils (paraffin and motor oil) were used to contaminate soil microcosms, and a simulated bioremediation treatment with nutrient-addition was applied. The indigenous microorganisms were monitored over 103 d using complementary community-level techniques (carbon source physiological profiling using Biolog® microplates, and phospholipid fatty acid (PLFA) profiling). Changes in the toxicity of the applied oils were monitored using luminescent bacterial bioassays, including Vibrio fischeri and a hydrocarbon-degrading Pseudomonas putida strain. Distinct shifts in microbial community structure and C source utilization profiles were observed as a result of oil contamination. There was some evidence that bioremediated soils were returning to control values by the end of the experiment. This was supported by the bioassay results which showed an initial increase in toxicity as a result of the oil addition which had then decreased by the conclusion of the experiment. The two oils exhibited markedly different toxicity towards the bioassay organisms, with species-specific differences in response. This oil-specific difference was also found in the PLFA profiles which showed the two oil types selected different microbial communities.