Because of the extreme conditions of the Deepwater Horizon (DWH) release (turbulent flow at 1500m depth and 5°C water temperature) and the sub-surface application of dispersant, small but neutrally buoyant oil droplets <70μm were formed, remained in the water column and were subjected to in-situ biodegradation processes. In order to investigate the biodegradation of Macondo oil components during the release, we designed and performed an experiment to evaluate the interactions of the indigenous microbial communities present in the deep waters of the Gulf of Mexico (GOM) with oil droplets of two representative sizes (10μm and 30μm median volume diameter) created with Macondo source oil in the presence of Corexit 9500 using natural seawater collected at the depth of 1100–1300m in the vicinity of the DWH wellhead. The evolution of the oil was followed in the dark and at 5°C for 64days by collecting sacrificial water samples at fixed intervals and analyzing them for a wide range of chemical and biological parameters including volatile components, saturated and aromatic hydrocarbons, dispersant markers, dissolved oxygen, nutrients, microbial cell counts and microbial population dynamics. A one phase exponential decay from a plateau model was used to calculate degradation rates and lag times for more than 150 individual oil components. Calculations were normalized to a conserved petroleum biomarker (30αβ-hopane). Half-lives ranged from about 3days for easily degradable compounds to about 60days for higher molecular weight aromatics. Rapid degradation was observed for BTEX, 2–3 ring PAHs, and n-alkanes below n-C23. The results in this experimental study showed good agreement with the n-alkane (n-C13 to n-C26) half-lives (0.6–9.5days) previously reported for the Deepwater Horizon plume samples and other laboratory studies with chemically dispersed Macondo oil conducted at low temperatures (<8°C). The responses of the microbial populations also were consistent with what was reported during the actual oil release, e.g. Colwellia, Cycloclasticus and Oceanospirillales (including the specific DWH Oceanospirillales) were present and increased in numbers indicating that they were degrading components of the oil. The consistency of the field and laboratory data indicate that these results could be used, in combination with other field and model data to characterize the dissipation of Macondo oil in the deepwater environment as part of the risk assessment estimations.