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
7
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
8
HYDROCARBON BIODEGRADATION
9
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
14
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
15
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
16
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
17
Source Indices
Hopanes
Steranes
18
BIODEGRADATION MOLECULAR MARKERS
Acyclic isoprenoids Diasteranes
C27-steranes bb-Steranes Homohopanes
Monoaroamtic steranes Triaromatic steranes
biodegradation
19
Effect of Biodegradation on Source Diagnostic
Ratios
Index Original oil Biodegraded oil Biodegraded oil
Index Original oil AM 20 days AMS200 40 days
Pr/Ph 0.98 0.97 ---
23ba 63 62 70
24TT 11 11 7
27Ts 22 21 22
29ab 43 43 42
27d 36 39 29
29aaS 45 49 49
29bb 47 49 57
27bb 37 37 7
isoprenoides
hopanes
steranes
20
Source Indices
D2/P2 D3/P3
21
Effect of Biodegradation on Source Diagnostic
Ratios
Index Original oil Biodegraded oil Biodegraded oil
Index Original oil AM 20 days AMS200 40 days
D2/P2 34 34 25
D3/P3 40 37 33
22
Weathering Indices
D2/C2 D3/C3
C2
256270
C3
D2
D3
23
Effect of Biodegradation on Weathering
Diagnostic Ratios
Index Original oil Biodegraded oil Biodegraded oil
Index Original oil AM 20 days AMS200 40 days
D2/C2 67 59 12
D3/C3 70 70 51
24
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.

25
ACKNOWLEDGEMENTS

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