Cleaning Oil Spills Through Bio Remediation

Becky Otter

March 3, 2006

Where and How Often Do Oil Spills Where and How Often Do Oil Spills Occur?Occur?

Last year alone there were 21 spills between 7-700 Last year alone there were 21 spills between 7-700 tonnes of oil and 3 spills over 700 tonnes of oil. tonnes of oil and 3 spills over 700 tonnes of oil.

(www.itopf.com/stats.html )

(www.itopf.com/stats.html )

~ 2 million gallons of bilge water is dumped per year into the oceans with an average of 100ppm of oil http://www.acnatsci.org/education/kye/hi/kye52002.html#sec3)

Making Refined OilMaking Refined OilStructure of “Heavy” Crude oil.

(http://www.lloydminsterheavyoil.com/whatislaunch.htm)

(www.eia.doe.gov/.../ Refining_text.htm)

Oil Distillation Column in Fawley UK

Clean Up MethodsClean Up Methods

Mechanical containment or recoveryMechanical containment or recovery Chemical and biological Chemical and biological PhysicalPhysical

What Affects How Diesel Fuel is What Affects How Diesel Fuel is Degraded by Microbes?Degraded by Microbes?

Hydrophobic nature of oil limits transfer to cell Hydrophobic nature of oil limits transfer to cell surfaces of micro-organismssurfaces of micro-organisms

Concentration of oilConcentration of oil Composition of oil Composition of oil TemperatureTemperature pH if the oil is on soilpH if the oil is on soil ViscosityViscosity

(Leahy and Colwell, Microbiol Rev. 1990 September; 54(3): 305–315Leahy and Colwell, Microbiol Rev. 1990 September; 54(3): 305–315 )(Lee, et al.Lee, et al., Journal of Applied Microbiology. 2006; 100: 325-333.)Journal of Applied Microbiology. 2006; 100: 325-333.)

R. erythropolis, a close relative to EN3 strain

How The Surfactant Interacts with How The Surfactant Interacts with the Hydrocarbonthe Hydrocarbon

(Makkar and Rockne, Enviro. Tox and Chem 2003)

The surfactant surrounds the hydrocarbon allowing the bacterial cell to take up the hydrocarbon

The hydrophobic tails of the surfactants interact which each other forming a micelle in an aqueous solution

““Enhanced biodegration of diesel oil by a newly identified Enhanced biodegration of diesel oil by a newly identified Rhodococcus Rhodococcus baikonurensisbaikonurensis EN3 in presence of mycolic acid” EN3 in presence of mycolic acid”

The EN3 strain can undergo the following processThe EN3 strain can undergo the following process

Mycolic acid (a surfactant) was synthesized to Mycolic acid (a surfactant) was synthesized to increase the bioavailability of diesel to microbesincrease the bioavailability of diesel to microbes

Scientists compared mycolic acid enhancement to Scientists compared mycolic acid enhancement to enhancement by other surfactants at 20,000mg/l of enhancement by other surfactants at 20,000mg/l of oil.oil.

Used gas chromatography to measure the amounts of Used gas chromatography to measure the amounts of diesel degradeddiesel degraded

(Lee, et al.Lee, et al., Journal of Applied Journal of Applied Microbiology. 2006; 100: 325-333.)Microbiology. 2006; 100: 325-333.)

Mycolic Acid

O2

CO2 H2O+

ANY HYDROCARBON

ResultsResults

Figure 4 Degradation of diesel oil by Rhodococcus baikonurensis strain EN3. Strain EN3 was inoculated into MS solutions with diesel oil at different concentrations: (a) 1000, (b) 5000, (c) 10 000, and (d) 20 000 mg l 1. Initial inoculated cell concentration was 6 ×104 CFU ml 1. •, Uninoculated cultures as controls; ○, inoculated cultures for the experiments.

Figure 5 Chromatograms of diesel oil degradation [(a) 0 day, (b) first day, (c) second day, (d) third day, (e) fourth day, (f) fifth day, (g) sixth day and (h) seventh day]. The added diesel oil concentration was 10 000 mg l 1. Two of the residual peaks at the final day (h) were identified as the recalcitrant hydrocarbons pristane (Pr) and phytane (Ph).

Figure 6Figure 6 Effect of surfactants (100 mg l 1) on  Effect of surfactants (100 mg l 1) on the degradation of diesel oil by the degradation of diesel oil by Rhodococcus baikonurensisRhodococcus baikonurensis strain EN3. strain EN3. Initial concentration of the diesel oil was Initial concentration of the diesel oil was 20 000 mg l 1, and the initial cell 20 000 mg l 1, and the initial cell concentration 6 × 104 CFU ml 1. concentration 6 × 104 CFU ml 1.

Figure 7 Effect of mycolic acid on the degradation of diesel oil by Rhodococcus baikonurensis strain EN3. Initial concentration of the diesel oil was 20 000 mg l 1, and the initial cell concentration 6 × 104 CFU ml 1. Initial concentration of mycolic acid: ○, 10 mg l 1; , 50 mg l 1; , 100 mg l 1. •, Uninoculated cultures without mycolic acid as control 1; , inoculated cultures without mycolic acid as control 2.

ResultsResults After 7 days EN3 alone degraded almost After 7 days EN3 alone degraded almost

Most of the degradation occurring in the first 3 days.Most of the degradation occurring in the first 3 days. Other surfactants can enhance degradation of diesel, Other surfactants can enhance degradation of diesel,

however mycolic acid is the most effectiveat however mycolic acid is the most effectiveat 20,000mg/l.20,000mg/l.

There was no real difference between maximum level There was no real difference between maximum level of degradation with samples containing mycolic acid at of degradation with samples containing mycolic acid at 10, 50, & 100 mg/l.10, 50, & 100 mg/l.

Amount of Diesel Fuel Percent degraded after 7 days

1000mg/l ~100%

5000mg/l 64.90%

10000mg/l 60.70%

20000mg/l 30.50%

ConclusionConclusion The bacterial strain EN3 along with synthetic mycolic The bacterial strain EN3 along with synthetic mycolic

acid has the potential to degrade fairly large amounts acid has the potential to degrade fairly large amounts of fuel over a short period of time in both aqueous and of fuel over a short period of time in both aqueous and soil environmentssoil environments

This is new alternative or additive to current methods This is new alternative or additive to current methods of cleaning oil spills without the other worries such as of cleaning oil spills without the other worries such as how to dispose of the diesel soaked sorbents or how to dispose of the diesel soaked sorbents or environmental effects caused from burning off the oilenvironmental effects caused from burning off the oil

SourcesSources Academy of Natural Sciences website, accessed Feb 2006. Academy of Natural Sciences website, accessed Feb 2006.

http://www.acnatsci.org/education/kye/hi/kye52002.html#sec3http://www.acnatsci.org/education/kye/hi/kye52002.html#sec3 Biswas, et al. “Microbial uptake of diesel oil sorbed on soil and oil Biswas, et al. “Microbial uptake of diesel oil sorbed on soil and oil spill clean-up spill clean-up

sorbents.” Journal of Chemical Technology and Biotechnology. 2005; 80: sorbents.” Journal of Chemical Technology and Biotechnology. 2005; 80: 587-587- 593.593.

Burford and Hien. “Effect of Foam Density, Oil Viscosity, and Burford and Hien. “Effect of Foam Density, Oil Viscosity, and Temperature Temperature on on Oil Sorption Behavior of Polyurethane.” Oil Sorption Behavior of Polyurethane.” Journal of Applied Polymer Journal of Applied Polymer Science. Science. 2006; 99: 360-367.2006; 99: 360-367.

ITOPF website, accessed Feb 2006.ITOPF website, accessed Feb 2006. (http://www.itopf.com/stats.html)(http://www.itopf.com/stats.html) Leahy and Colwell.“Microbial degradation of hydrocarbons in the environment.” Leahy and Colwell.“Microbial degradation of hydrocarbons in the environment.”

Microbiol Rev. 1990 September; 54(3): 305–315 Microbiol Rev. 1990 September; 54(3): 305–315 Lee, et al. “Enhanced biodegration of diesel oil by newly identified Lee, et al. “Enhanced biodegration of diesel oil by newly identified Rhodococcus Rhodococcus

baiknurensisbaiknurensis EN3 in the presence of mycolic acid.” Journal of Applied EN3 in the presence of mycolic acid.” Journal of Applied Microbiology. 2006; 100: 325-333.Microbiology. 2006; 100: 325-333.

Makkar R. and Rockne K. “Comparison of synthetic surfactants and biosuractants in Makkar R. and Rockne K. “Comparison of synthetic surfactants and biosuractants in enhancing biodegration of polycyclic aromatic hydrocarbons” enhancing biodegration of polycyclic aromatic hydrocarbons”

Environmental Environmental Toxicology and Chemistry. 2003; 22 (10): 2280-2292.Toxicology and Chemistry. 2003; 22 (10): 2280-2292. Oil Spills-EPA website, accessed Feb 2006 Oil Spills-EPA website, accessed Feb 2006 ((

http://www.epa.gov/oilspill/oiltech.htmhttp://www.epa.gov/oilspill/oiltech.htm). ).