Centre for Marine Biodiversity and Biotechnology Open Day 2012. Presentation by Tony Gutierrez.

1. Revealing the Identity of
Oil-Degrading Microorganisms
at Deepwater Horizon
….and a little more

2. Overview
1. Deepwater Horizon oil spill in the GoM.
Where did most of the oil go? Who’s responsible?
2. On the formation of ‘Oily Snot’ during DWH spill.
3. FISH’n for oil-degrading bacteria.
4. Knowns and Unknowns about the DWH spill.

3. Deepwater Horizon Mucondo well site

4. Identifying Who Degraded the Oil by Stable-Isotope Probing (SIP)
A
Uniformly-labeled trans-2-hydroxy- Incorporation of the 13C
13C Naphthalene benzylidenepyruvate atoms into cellular biomass
(e.g. DNA)
Degradation OH Further metabolism
COO-
“ Heavy ” DNA
O
Incubation of sample on
B labeled 13C PAHs
Whole DNA
“ light ”
extract
Assimilation of label - PCR of 16S rRNA
by active microbes DNA - Cloning
- Sequencing
“ heavy ”

5. Gutierrez, T. (2011) Rev Environ Sci Bio/Technol.

6. What about that ‘Oily Snot’
A
Hydrocarbon concentration (mg/L)
50
40
30
20
10
0
0.0 0.2 0.4 0.6 0.8 1.0
Polymer concentration (mg/mL)
B1
10 mm
500
2
400
300
Percent change
PE5
200
7
100
0
0 0
-100
B6 B11

7. FISH’n for Oil-Degrading Bacteria
( Fluorescence In Situ Hybridization )
G-T-G-C-C-G-C-T-A-T-C-G-A-A-T-T-C-G

8. Alpha-proteobacteria associated with Pseudonitzschia diatom
Probes used: EUB-Cy5 / Alpha-Cy3

9. Gulf of Mexico Plume Water Enriched with Oil
Beginning of enrichment 4 days later
Marinobacter (green/yellow)
Marinobacter (green) + DAPI (blue) vs. others (red)

10. Overview about the DWH Oil Spill:
a microbial perspective
Knowns:
- Mobility / Fate of the oil
- Bacteria largely responsible in ultimate removal of the oil
- The types of bacteria that dominated and were likely
responsible for degrading much of the oil
- We know that all of the oil is now gone from the water
column; but residual oil remains in the sediment.
Not-So-Well Knowns:
- Detail on the microbial response and succession of this response
over the course of the spill
- What diversity of bacteria contributed, and by how much?
What was their function?

11. Acknowledgements
Colleagues/Collaborators
• Prof. Mike Aitken (UNC, Chapel Hill)
• Prof. Andreas Teske (UNC, Chapel Hill)
• Dr. Kirk Semple (Univ. Lancaster)
• Dr. Alexander Loy (Univ. Vienna, Austria)
• Luke McKay (UNC, Chapel Hill)
• Tingting Yang (UNC, Chapel Hill)
• Sara Mishamandani (UNC, Chapel Hill)
Funders
• European Commission (Marie Curie Programme)
• NSF Rapid Response Grant
• UNC fellowship program