Biodegradation of the Prestige fuel oil under simulated
Biodegradation of the Prestige fuel oil under simulated conditions S. DÍEZa, J. SABATÉb, M. VIÑASb, J. M. BAYONAa, A. M. SOLANASb and J. ALBAIGÉSa Instituto Investigaciones Químicas y Ambientales de Barcelona-CSIC. B Universidad de Barcelona. Departamento de Microbiología a
The fate of spilled oil in the marine environment Fuente: Albaigés 1989
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.
METHODOLOGY
Microbial Consortia Enrichment Inocula: crude oil contaminated sand + 5 -PAHs (3 -4 aromatic ring) AM 5 years Mineral medium Inocula: diesel contaminated Soil + diesel oil TD
Biodegradation Experiments S 200 : microemulsion urea + oleic acid + phosphate ester +2 -butoxy-1 -ethanol original fuel AM 50 mg TD 50 mg weathered fuel AM 50 mg TD 50 mg fuel + S 200 AM 50 mg TD 50 mg 40 days 20 days 50 mg original fuel Chemical analysis
Chemical Analysis Incubated fuel surrogates p. H < 2. 5 LLE (CH 2 Cl 2) organic extract Fractionation SPE (Si. O 2 -C 3 CN) Aliphatics GC-FID GC-MS Aromatics GC-MS
HYDROCARBON BIODEGRADATION
Biodegradation Aliphatics 22 15 100 original fuel 90 80 x 70 60 Ph 50 x x abundance 40 Pr x 32 30 20 10 100 90 biodegraded fuel 20 days 80 70 60 50 Pr Ph 40 30 x 20 x x x 10 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 Time (min)
Biodegradation Aromatics 40 N 1 35 original fuel N 2 30 N 3 25 20 P P 1 P 2 15 P 3 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 Time (min) 26 28 30 32 34 36 38 40 42 44
Weathering Effect
Strain Efficiency
Oleophilic Nutrient Effect
Alkylbenzenes 100 (II) ORIGINAL FUEL m/z 106 alkyltoluenes 80 60 40 20 0 100 Relative Abundance 80 1 -methyl-3 -phytanylbenzene 60 (II) BIODEGRADED (TD) m/z 106 40 20 0 100 80 ORIGINAL FUEL 60 m/z 92 alkylbenzenes 40 (I) 20 0 100 phytanylbenzene 80 (I) BIODEGRADED (TD) 60 m/z 92 40 20 0 6 8 10 12 14 16 18 20 22 24 Time (min) 26 28 30 32 34 36 38
C 2 & C 3 -Phenanthrenes 100 C 2 -Phenanthrenes (P 2) 80 ORIGINAL m/z 206 60 40 20 Relative Abundance 0 100 80 BIODEGRADED 60 m/z 206 40 20 0 100 C 3 -Phenanthrenes (P 3) 80 60 ORIGINAL m/z 220 40 20 0 100 80 BIODEGRADED 60 40 m/z 220 20 0 16, 5 17, 0 17, 5 18, 0 18, 5 19, 0 19, 5 Time (min) 20, 0 20, 5 21, 0 21, 5 22, 0 22, 5 23, 0
C 2 & C 3 - Dibenzothiophenes 100 C 2 - Dibenzothiophenes (D 2) 80 60 ORIGINAL m/z 212 40 20 0 100 Relative Abundance 80 BIODEGRADED 60 m/z 212 40 20 0 100 80 C 3 - Dibenzothiophenes (D 3) 60 40 ORIGINAL m/z 226 20 0 100 80 BIODEGRADED 60 m/z 226 40 20 0 13, 5 14, 0 14, 5 15, 0 15, 5 16, 0 16, 5 17, 0 17, 5 Time (min) 18, 0 18, 5 19, 0 19, 5 20, 0 20, 5 21, 0 21, 5
Source Indices Hopanes Steranes
biodegradation BIODEGRADATION MOLECULAR MARKERS Acyclic isoprenoids Diasteranes C 27 -steranes bb-Steranes Homohopanes Monoaroamtic steranes Triaromatic steranes
steranes hopanes isoprenoides Effect of Biodegradation on Source Diagnostic Ratios Biodegraded oil Index Original oil AM 20 days AM+S 200 40 days Pr/Ph 0. 98 0. 97 --- %23 ba 63 62 70 %24 TT 11 11 7 %27 Ts 22 21 22 %29 ab 43 43 42 %27 d 36 39 29 %29 aa. S 45 49 49 %29 bb 47 49 57 %27 bb 37 37 7
Source Indices D 2/P 2 D 3/P 3 100 90 80 P 2 70 60 P 3 50 206+220 40 30 20 100 D 2 90 D 3 80 70 212+226 60 50 40 30 20 10 0 14, 5 15, 0 15, 5 16, 0 16, 5 17, 0 17, 5 18, 0 18, 5 19, 0 19, 5 Time (min) 20, 0 20, 5 21, 0 21, 5 22, 0 22, 5 23, 0 23, 5 24, 0
Effect of Biodegradation on Source Diagnostic Ratios Biodegraded oil Index Original oil AM 20 days AM+S 200 40 days D 2/P 2 34 34 25 D 3/P 3 40 37 33
Weathering Indices D 2/C 2 D 3/C 3 C 2 100 90 256+270 80 C 3 70 60 50 40 30 20 100 D 2 90 D 3 80 212+226 70 60 50 40 30 20 10 0 13 14 15 16 17 18 19 20 21 22 23 Time (min) 24 25 26 27 28 29 30 31 32
Effect of Biodegradation on Weathering Diagnostic Ratios Biodegraded oil Index Original oil AM 20 days AM+S 200 40 days D 2/C 2 67 59 12 D 3/C 3 70 70 51
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.
ACKNOWLEDGEMENTS • Financial support was obtained from the MCYT (VEM 2003 -20068 -CO 5 -01). • Dr. S. Diez acknowledges a Ramon & Cajal contract.
- Slides: 25