Biodegradation of the Prestige fuel oil under simulated conditions

1
Biodegradation of the Prestige fuel oil under
simulated conditions

• S. DÍEZa, J. SABATÉb, M. VIÑASb, J. M. BAYONAa,
  A. M . SOLANASb and J. ALBAIGÉSa
• a Instituto Investigaciones Químicas y
  Ambientales de Barcelona-CSIC.
• B Universidad de Barcelona. Departamento de
  Microbiología

2
The fate of spilled oil in the marine environment
Fuente Albaigés 1989
3
OBJECTIVES

• To evaluate the biodegradation extent of Prestige
  fuel oil in vitro experiments times by two
  consortia.
• Aliphatics, aromatics and molecular markers.
• Effect of biodegradation on
• source and weathering indices
• To assess weathering effects on biodegradation.
• To evaluate the effect of oleophilic nutrients.

4
METHODOLOGY
5
Microbial Consortia Enrichment
Mineral medium
Inocula crude oil contaminated sand
5-PAHs (3-4 aromatic ring)
Inocula diesel contaminated Soil diesel oil
5 years
AM
TD
6
Biodegradation Experiments
S200 microemulsion urea oleic acid
phosphate ester 2-butoxy-1-ethanol
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Chemical Analysis
Incubated fuel
pH lt 2.5 LLE (CH2Cl2)
surrogates
organic extract
Fractionation SPE (SiO2-C3CN)
Aliphatics GC-FID GC-MS
Aromatics GC-MS
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HYDROCARBON BIODEGRADATION
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Biodegradation Aliphatics


10
Biodegradation Aromatics


40

N1

original fuel
35
N2


30

N3

25

20

P1

P2

P

15

P3

N

10

5


0
40

35
biodegraded fuel 20 days

30

25

20

15

10

5

0























2
4
6
8
10
12
14
16
18
20
22
24
26
28
30
32
34
36
38
40
42
44
Time (min)
11
Weathering Effect
12
Strain Efficiency
13
Oleophilic Nutrient Effect
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Alkylbenzenes
(II)
ORIGINAL FUEL
m/z 106 alkyltoluenes
(II)
BIODEGRADED (TD)
1-methyl-3-phytanylbenzene
m/z 106
ORIGINAL FUEL
m/z 92 alkylbenzenes
(I)
phytanylbenzene
(I)
BIODEGRADED (TD)
m/z 92
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C2 C3-Phenanthrenes
C2-Phenanthrenes(P2)
ORIGINAL
m/z 206
BIODEGRADED
m/z 206
C3-Phenanthrenes(P3)
ORIGINAL
m/z 220
BIODEGRADED
m/z 220
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C2 C3- Dibenzothiophenes
C2- Dibenzothiophenes (D2)
ORIGINAL
m/z 212
BIODEGRADED
m/z 212
ORIGINAL
C3- Dibenzothiophenes(D3)
m/z 226
BIODEGRADED
m/z 226
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Source Indices
Hopanes
Steranes
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BIODEGRADATION MOLECULAR MARKERS
Acyclic isoprenoids Diasteranes
C27-steranes bb-Steranes Homohopanes
Monoaroamtic steranes Triaromatic steranes
biodegradation
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Effect of Biodegradation on Source Diagnostic
Ratios
isoprenoides
hopanes
steranes
20
Source Indices
D2/P2 D3/P3
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Effect of Biodegradation on Source Diagnostic
Ratios
22
Weathering Indices
D2/C2 D3/C3
C2
256270
C3
D2
D3
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Effect of Biodegradation on Weathering
Diagnostic Ratios
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CONCLUSIONS

• N-alkanes, alkylbenzenes low MW parent PAHs and
  mono and di-alkylated derivatives are degraded by
  both consortia in 20-40 days incubation.
• Degradation rates of PAHs decrease according with
  the alkyl substitution and number aromatic rings.
• Weathering slow down biodegradation.
• Oleophilic nutrient enhance the biodegradation of
  the more refractory PAHs and some molecular
  markers.
• Source indicators can be modified when oleophilic
  nutrient are added to the incubation medium.

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ACKNOWLEDGEMENTS

• Financial support was obtained from the MCYT
  (VEM2003-20068-CO5-01).
• Dr. S. Diez acknowledges a Ramon Cajal contract.