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Oil Spill Bioremediation

[Your Name]

SCI 207: Our Dependence Upon the Environment

[Instructor’s Name]

[Date]
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Oil Spill Bioremediation

Annually, millions of gallons of oil enter the seas. The poisoning of marine ecosystems

with oil has long-lasting repercussions. The majority of ocean oil pollution is caused by smaller,

less-noticed incidents, such as spills from smaller oil ships, the routine operation of oceanic oil

wells, leaking storage tanks and piping for offshore oil wells, and poorly drilled holes on the

ocean floor (Panchal et al., 2018). The oil may be released into the water when loading and

unloading ships. On land and in waterways, refined oil from municipal and industrial sources is

frequently abandoned, spilled, or leaked. Heavy rains send road oil to streams and the ocean, and

several people illegally dispose of used motor oil. This oil can reach the ocean via storm drains,

rivers, and streams. Smoke from oil burns can also enter the atmosphere and precipitate into the

ocean and streams as rain. The toxic chemicals can kill plankton and algae in the oil. Fish that

consume contaminated food may become ill or die. Eliminating, infecting, or deforming fish

larvae decreases future populations (Pete et al., 2021).

The toxicity of the oil kills or permeates these creatures, turning them toxic. Oil coats the

feathers and hair of birds and aquatic animals, decreasing their insulation, buoyancy, and

mobility. The majority of natural seepage from marine oil deposits is absorbed by ocean-

dwelling bacteria that have developed pathways to utilize oil as a source of food and energy

(Bhattacharya et al., 2019). Typically, microorganisms biodegrade long-chain hydrocarbons

from petroleum into energy and nutrients for their biological functions. Hydrocarbons enable

organisms to produce biomass and thrive. Microorganisms that degrade oil collaborate to break

down oil. Usually, the hydrocarbon ring structures are broken down into carbon dioxide by

marine bacteria and fungi using saltwater enzymes and oxygen. The goal of this experiment was

to mimic marine oil spill bioremediation by utilizing oil-eating microbes. The degradation of oil
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will be monitored using Tetrazolium as an indicator. When oxidized, tetrazolium is usually

colorless, but it turns pink when its chemical makeup is altered. Breaking down oil carbon

compounds releases electron donors as byproducts of the process. The addition of hydrogen to

the tetrazolium indicator changes the chemical makeup of the indicator, causing it to become

pink (due to the creation of an insoluble pink compound). In this experiment, tetrazolium was

reduced from its oxidized, colorless state to its reduced, pink state, signifying that oil was being

broken down.

Hypotheses

Activity 1:

Tubes 1 and 2

Tube 2 will change color. There will be microbes in tube 2, which will break down the

oil. Test tube one will not have these microbes, and there will be no color change.

Tubes 3 and 4

There will be visible changes in color in test tube 4. The microbes in tube four will

metabolize, thus resulting in a change in physical appearance.

Tubes 5 and 6

Tube 5 will undergo less oil breakdown than tube 6. More microbes added in test tube six

will result in an increased breakdown. The color change will also be vigorous in test tube six.

Materials and Methods

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Various materials were required for the experiment. These included: Rid-X Septic

System Treatment (powder), 20g, ten plastic pipets, 3 mL, graduated, 6 Culture tubes with caps,

graduated cylinder, 2 Plastic cups, Bag containing tetrazolium indicator powder (0.02 g), and

distilled water (100 mL), 0.02%, Cheesecloth, funnel, test tube rack, cooking oil, bottled water,

30mL, warm tap water, 140 mL, 10% bleach solution for cleanup, pencil, stopwatch, and a

camera.

Activity 1: Bioremediation of Oil

Using a pencil, the culture tubes 1 through 6 were labeled. Tubes 1 and 2 employed a

tetrazolium indicator to determine whether or not metabolism occurred. In tubes 3 and 4, the

appearance and physical characteristics of the oil in the presence of microorganisms were

evaluated. In tubes 5 and 6, the influence of the selected environmental factors on the oil

breakdown by microorganisms was examined. As environmental parameters change, the quantity

of bacteria present, the amount of oil present, the kind of oil present, the light conditions, or the

variance in temperature change. Using a plastic pipet, 1 mL of 0.02 percent tetrazolium indicator

was pipetted into tubes 1, 2, 5, and 6. Only tubes 1, 2, 3, and 4 received 2 mL of distilled water

added using a clean plastic pipet. Ten drops of oil were applied using a clean plastic pipet in each

of the six tubes. Using a clean plastic pipet, two milliliters of distilled water were added to tube

1, three milliliters to tube 3, and one milliliter to tube 4. Only tubes 2, 4, and 6 received 2 mL of

microbial suspension using a clean plastic pipet. All of the culture tubes were sealed, and the

liquid in each of the six tubes was mixed by finger vortexing. The vortexing created a whirlpool

inside the tube, allowing for the mixing of the liquids. This procedure was done with the other

test tubes while all six tubes were positioned vertically in a test tube rack.
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Results
Observations

Data Table 1. Tubes 1 and 2 Observations

Day Tube 1 Tube 2

Clear liquid; settled oil on the top visible Light cream-colored fogginess
0
(Initial setup)

Clear; oil still present Fogginess gets stronger. Oil breaks down.
1
No changes Still foggy, oil continues to break down.
2
No changes Still very foggy, oil is almost fully broken
3 down
Data Table 2. Tubes 3 and 4 Observations

Day Tube 3 Tube 4

Presence of clear oil -the clearer the tube 1. Oil is visible. However, there is light
0 fogginess but still clearer than in tube 2,
(Initial setup)
Oil starts to break down but is still visible Oil is more broken down than tube 3
1
Oil is not as separated as on day 0. Breaking down of oil intensifies. Fogginess
2 is stronger.
Oil is still visible but not as much as tube 1. Oil has broken down, but not fully;
3 fogginess persists.

Data Table 3. Tubes 5 and 6 Observations

Chosen environmental factor in changing: Placing test tube 5 in the dark

Day Tube 5 Tube 6

Oil is visible; however, the tube is the Almost the same as tube 5, a little bit
0 foggiest due to the extra microbial lighter than tube 5.
(Initial solutions.
setup)
Breaking down of oil is very fast; still The color is lighter than that of tube 5 –
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1 froggy. color almost identical to that of tube 5

Oil is invisible; the extra microbial solution Darkening continues. Oil slightly visible
2 seems to have eaten away all oil.
No traces of oil, foggiest tube. No oil present, quite dark
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Discussion

From the results obtained, all three of my hypotheses. By day two, tube two had already

begun to change color. It was evident that the oil had deteriorated and changed color. For the

second hypothesis, tube four had some oil separation, indicating a change in physical

characteristics. Lastly, oil decomposition was accelerated by increasing the number of

microorganisms in tube six. Bioremediation is an excellent approach to achieving environmental

sustainability. In the event of a significant oil spill in a large body of water, these

microorganisms can aid in the degradation of these spillage zones. The bacteria convert

hydrocarbons, such as oil, into harmless byproducts, thereby reducing the danger to creatures.

While breaking down these oils, bacteria alter the physical characteristics that make the oil film

more malleable and breakable. With these features, marine life will be less likely to suffocate.

Therefore, microorganisms are a reliable means of mitigating the impact of oil spills in maritime

settings.

I have learned that fast biodegradation of oil occurs when the oil is widely spread in the

water column, and the microbes are well-adapted to hydrocarbon exposure, as in Gulf of Mexico

waters (Bhattacharya et al., 2019). Bioremediation with fertilizer can speed up the

biodegradation of oil in specific circumstances. However, biodegradation will not remove all of

the oil from the environment; patches of weathered oil are likely to persist in certain areas.

Detectable hydrocarbons should not be the sole determining factor in whether or not to use
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microbial oil biodegradation, including bioremediation. A variety of factors influence an oil

spill's outcome, including the amount of oil released, the chemical composition of that oil, and

the ecosystems affected by the spill's effects on those ecosystems' particular environmental

conditions (Bhattacharya et al., 2019). However, the worldwide character of oil-degrading

microorganisms is a common denominator.

The major problem with the experiment was the time it took. The lab took three days,

making it difficult to follow up on the experiment promptly. This could have led to obtaining

inconsistent or undesired results. Also, students were expected to conduct the lab on their own.

Any failure to follow/misunderstand the instructions to the latter could result in erroneous

results. Poor observations of the color changes could also result in making wrong inferences.

Lastly, incorrect measurements of the components required lead to a slow or extremely faster

reaction, thus making wrong inferences.

Future research should major on what responders of oil spillage can reduce to prevent the

oil from entering seas and oceans. There is a need for a large-scale and faster scientific response

to oil spillage. Frontline responders to oil spillage should be prepared to quickly mobilize a

scientific understanding of the particular conditions at hand, such as natural and increased

biodegradation, and establish the best feasible way to minimize the danger and environmental

effect of a future spill.

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References

Bhattacharya, M., Guchhait, S., Biswas, D., & Singh, R. (2019). Evaluation of a microbial

consortium for crude oil spill bioremediation and its potential uses in enhanced oil

recovery. Biocatalysis and agricultural biotechnology, 18, 101034.

Panchal, A., Swientoniewski, L. T., Omarova, M., Yu, T., Zhang, D., Blake, D. A., ... & Lvov,

Y. M. (2018). Bacterial proliferation on clay nanotube Pickering emulsions for oil spill

bioremediation. Colloids and Surfaces B: Biointerfaces, 164, 27-33.

Pete, A. J., Bharti, B., & Benton, M. G. (2021). Nano-enhanced bioremediation for oil spills: a

review. ACS ES&T Engineering, 1(6), 928-946.