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This study evaluates the extent of biodegradation of Prestige fuel oil under simulated conditions. The effects of biodegradation on aliphatics, aromatics, and molecular markers are assessed, as well as the impact of weathering and oleophilic nutrients. The results indicate that certain compounds are degraded by microbial consortia, and weathering can slow down biodegradation. The addition of oleophilic nutrients enhances the degradation of more refractory compounds. This research provides insights into the fate of spilled oil in marine environments.
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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
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.
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
original fuel original fuel weathered fuel fuel + S200 50 mg AM 50 mg TD 50 mg AM 50 mg TD 50 mg AM 50 mg TD 50 mg 20 days 40 days C h e m i c a l a n a l y s i s C h e m i c a l a n a l y s i s Biodegradation Experiments S200 : microemulsion urea + oleic acid + phosphate ester +2-butoxy-1-ethanol
Chemical Analysis Incubated fuel pH < 2.5 LLE (CH2Cl2) surrogates organic extract Fractionation SPE (SiO2-C3CN) Aliphatics GC-FID GC-MS Aromatics GC-MS
22 15 original fuel 100 90 80 x 70 60 Ph x Pr 32 50 x x 40 30 20 abundance 10 1 00 90 biodegraded fuel 20 days 80 70 60 50 Pr Ph 40 x x 30 x x 20 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 Aliphatics
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)
100 80 60 40 20 0 100 80 60 40 20 Relative Abundance 0 100 80 60 40 20 0 100 80 60 40 20 0 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 Time (min) 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
100 80 60 40 20 0 100 80 60 Relative Abundance 40 20 0 100 80 60 40 20 0 100 80 60 40 20 0 16,0 16,5 17,0 17,5 18,0 18,5 19,0 19,5 20,0 20,5 21,0 21,5 22,0 22,5 23,0 Time (min) 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
100 80 60 40 20 0 100 80 60 Relative Abundance 40 20 0 100 80 60 40 20 0 100 80 60 40 20 0 13,5 14,0 14,5 15,0 15,5 16,0 16,5 17,0 17,5 18,0 18,5 19,0 19,5 20,0 20,5 21,0 21,5 Time (min) C2 & C3- Dibenzothiophenes C2- Dibenzothiophenes (D2) ORIGINAL m/z 212 BIODEGRADED m/z 212 C3- Dibenzothiophenes(D3) ORIGINAL m/z 226 BIODEGRADED m/z 226
Source Indices Hopanes Steranes
BIODEGRADATION MOLECULAR MARKERS Acyclic isoprenoids Diasteranes C27-steranes bb-Steranes Homohopanes Monoaroamtic steranes Triaromatic steranes biodegradation
Effect of Biodegradation on Source Diagnostic Ratios isoprenoides hopanes steranes
100 90 80 P2 70 60 P3 50 206+220 40 30 20 10 0 100 D2 90 D3 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 20,0 20,5 21,0 21,5 22,0 22,5 23,0 23,5 24,0 Time (min) Source Indices D2/P2 D3/P3
100 90 80 70 60 50 40 30 20 10 0 100 90 80 70 212+226 60 50 40 30 20 10 0 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 Time (min) Weathering Indices D2/C2 D3/C3 C2 256+270 C3 D2 D3
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 (VEM2003-20068-CO5-01). • Dr. S. Diez acknowledges a Ramon & Cajal contract.