Patel: Effect of Temperature on Bi
3
Effect of Temperature on Bioremediation
Microbiology
_______________________________________________
Signature of Sponsoring Teacher
_______________________________________________
Signature of School Science Fair Coordinator
Khusbu Patel
2501 West Addison Street
Lane Tech College Prep High School
Chicago, IL 60618
Grade 10
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Table of Contents
Acknowledgements………………………………………………………………………………..5
Purpose…………………………………………………………………………………………….6
Hypothesis…………………………………………………………………………………………6
Review of Literature………………………………………………………………………………7
Materials...…………………………………………………………………………………….…18
Procedure…………..………………………………………………………………………….…19
Results……………………………………………………………………………………………20
Conclusion……………………………………………………………………………………….25
Reference List……………………………………………………………………………………27
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Acknowledgments
I would like to thank my chemistry teacher, Mr. Kopack, for ordering my materials. I
would also like to thank Dr. Lang for assisting me in my procedure and answering any questions
that I had about my experiment. I would also like to take this time to express my gratitude to
Mrs. Mikbel for proofreading and revising my entire paper, along with helping me understand
how to write an effective science paper. Mr. McAdam was one of the other teachers that helped
me create graphs and tables for the data that was collected during my experiment. In addition, I
would like to extend my appreciation to my dad for buying some of the supplies necessary for
this experiment.
Patel: Effect of Temperature on Bi
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Purpose
The purpose of this experiment was to compare the amount of motor oil that is
biodegraded by Escherichia coli in different temperatures and to figure out if temperature affects
the biodegradation of motor oil.
Hypothesis
If motor oil and Escherichia coli are placed in 2 different temperatures, then the motor oil
placed in 37oC, the highest temperature, will have degraded the most because warmer
temperatures allow bacteria to grow, while colder temperatures freeze bacteria so that they do
not grow or work properly, causing less oil to be degraded.
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Review of Literature
Scientists have been using microbiology, the study of unicellular and multi-cellular
microscopic organisms, for many years now. The study of microbiology includes learning about
fungi and protists, also known as eukaryotes, and prokaryotes, which are organisms that do not
include a cell nucleus in their cells (Boyd, 1988). Also, viruses and prions are not classified as
living organisms, but they are still studied under the subject of microbiology (1988). Generally,
microbiology is the study of life that cannot be seen by the naked eye. The study of microbes has
helped the world in so many ways. Scientists have found information about illnesses and cures
for certain diseases. They also know how the human body works and how organisms affect
humans in various ways. In addition, microbiology has helped the environment in ways that
would not have been found if experts did not do research or experiments on theories and other
beliefs.
One area of research in microbiology deals with oil pollution, one of the most harmful
types of pollution on earth. It has also become an enormous problem in the world. Not only does
it harm humans, but oil can also hurt other types of animals as well. Birds, for example, can be
killed just by getting oil into their feathers, and as a result of this, the animals cannot stay warm,
so they freeze to death. In addition, the damage oil pollution causes is extremely hard to repair
and only certain procedures can help reduce oil pollution; one of them is using bacteria to
degrade the oil. The study of microbiology has helped scientists figure out that microorganisms
can remove human waste products from the environment. Different temperatures in the
environment, though, can cause different results to occur (Owens, 2004).This experiment is
testing if different temperatures can change the effect of microorganisms helping the
environment by getting rid of small amounts of oil, and, according to the research, it is highly
Patel: Effect of Temperature on Bi
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likely that microbes will be able to degrade oil in room temperature and in warmer climates, then
in colder temperatures.
Oil Spills
General effects of oil spills. All around the world, oil is being transported for human use.
Throughout this procedure, accidents usually happen which cause oil spills to occur. These spills
may sound like any normal, minor spill, but they are actually one of the most harmful types of
pollution caused by humans. Oil pollution has become a big problem ever since men started to
transport oil from one place to another (B. Delille, D. Delille, & Pelletier, 2002, p. 118-119).
Industrial activities, which include oil and gasoline exploration and transportation, contain toxic
waste, which is released into the environment because of the spills and accidents (Van der Meer,
2006, pp. 36-37). Oil spills can occur on land and even in the water. Oil is a hazardous chemical
used by humans for fuels, plastics, dyes, and many other substances. This substance is extremely
sticky and can damage the environment easily. Animals of all sorts, like birds, mammals, and
fish, can be killed just by ingesting oil. Others can be killed instantly from consuming a prey that
has been contaminated with oil (National Wildlife Federation, 1997, p. 57).
Cleaning up oil spills. Since oil is toxic, it is extremely difficult to clean up. Experts use
protective clothing and advanced equipment to protect them from coming into close contact with
the oil (1997). People have also started to become prepared for oil spills to occur by training
workers, improving technology, and plotting out marshes and endangered species’ habitats
(1997). This allows them to be ready for an oil spill when it happens. In addition, people have
tried to prevent oil spills by using safer navigation and screening a ship’s safety record to make
sure less and less oil spills occur each year (1997, p.88-89).
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Even though oil is a harmful substance, it does not mean it cannot be cleaned up. Oildegrading microorganisms can help clean up an oil spill. They first open up their genes that are
required for degradation (Van der Meer, 2006, p. 36-37). They can also increase their number in
size to the amount of carbon exposed (2006). After this, the microorganisms will take in the oil,
like they take in nutrients (2006, pp. 36-37). Another way for experts to quicken the process of
cleaning up is by adding fertilizer and oil decomposers to the oil spill (Fujita et al., 2003, pp.
442-443). Also, there are three different types of chemicals that can speed up the process of clean
oil spills: nutrients, cleaners, and dispersants (Nyman, 1999, para. 6). Nutrients can be added to
the oil spill to quicken the process of degradation of oil. Cleaners can be used to help clean oil
because they wash away the oil from surfaces back into the water, thus keeping land clean and
healthy (1999). Dispersants can be used because they help break the oil into smaller particles, or
droplets, allowing it to become easier to dilute the oil (1999). No response to oil spills is also
another option because the oil will eventually evaporate and degrade into wetlands, where the
soil is saturated with moisture (1999).
Bioremediation
Bioremediation is the removal or transformation of contaminants from the environment
by organisms (Litchfield, 2005). In other words, bioremediation of infected or polluted
environments is a way to restore the earth’s surface (Abraham et al., 2003, pp. 162). This method
has been going on for as long as humans had to degrade waste (Litchfield, 2005). Of course,
bioremediation is being used more often right now because better technology has been
developed. For instance, when oil is being transported from place to place, oil treatment agents,
like fertilizers, dispersants, and oil decomposers, are developed for oil spill bioremediation
Patel: Effect of Temperature on Bi
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(2005). For example, during the most recent oil spill, the Exxon Valdez, the method of
bioremediation was used to clean up the oil from the sea (Fujita et al., 2003, p. 442-443).
In addition, many tests and experiments have occurred to understand how bioremediation
works. For example, on June 8, 1990, the Norwegian tanker, Mega Borg, was carrying crude oil
about 57 miles off the Texas coast (Lee et al., 1996). All of a sudden, an explosion and fire
occurred, spilling approximately 45 m3 of Angolan Palanca crude oil into the ocean (1996). For
experimental purposes, a trial was conducted a day after the accident to test the bioremediation
of the oil. During the experiment, little of the oil was cleared up, but not much (1996). There
have been several other oil spill incidents where bioremediation products have been used in an
attempt to improve oil biodegradation. In general, though, it is difficult to draw valid conclusions
from many of these efforts because of the time constraints in planning experiments with the
appropriate controls after a major spill. Another type of field study was taken in February 1996,
and it is proved that, during the experiment, microbial response and activity was high and rapid
showing that the rate of degradation improves when bioremediation agents are present (Delille,
et al., 2002, p. 118-119).
Over the past few years, more and more people have become interested in developing a
cost-effective technique for bioremediation (2002, p. 118-119). Terry Hazen, the head of the
ecology department at the Lawrence Berkeley National Laboratory in California even said,
“Bioremediation holds great promise for some of our worst problems” (Voigt, 2010). Even
though, after many different trials, it is still proven that there is little convincing evidence to
suggest that bioremediation is effective, bioremediation does help remove waste in some way,
thus making the earth a better place to live in (Lee et al., 1996).
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Microorganisms
Wide-ranging microbes. Microorganisms, the simplest organisms in life, are able to
degrade any human waste products, including oil spills, making the earth a safer and cleaner
place to live in (Van der Meer, 2006, p. 35-36). All around the earth, millions and millions of
microorganisms exist. Even with that, though, scientists still believe that less than one percent of
all microorganisms have been identified (Voigt, 2010). Microbes represent about 50 to 90
percent of all life in the ocean (2010). In 1981, the very first microorganism to be patented was a
strain of the bacteria, Pseudomonas, which was found to degrade oil of four species in only one
(2010). The world’s oceans also contain microscopic life. There have been magnificent
discoveries of previously unknown microorganisms, most of which made a significant impact on
the earth (Arrieta et al., 2006, p. 12115-12116).
Bioremediation. The method microorganisms use to help and improve the environment is called
bioremediation. They detoxify and degrade the environmental contaminants, making them safer and less perilous
(Delille, et al., 2002, p. 118-126). When microorganisms are introduced to waste products, they learn to adapt to the
pollutants and then they start the degradation process (2002).
Microorganisms are crucial for an ecosystem to function. They are helpful in so many ways. For instance,
they purify water, soil, and even air (Van der Meer, 2006, p. 35-36). People rely on microorganisms to destroy waste
products from industrial, agricultural, and human domestic activities (2006). During this process, microorganisms
convert these wastes into carbon dioxide, water, and other biomasses (Litchfield, 2005, p. 273-279).
Microorganisms can also produce hydrogen from biodegration, and they can also make metals become less toxic
(2005). For example, microorganisms have been found to reduce choromium to the less hazardous type of
choromium, creating the use of metals that are safer and environmentally friendly (2005).
They are able to degrade objects that are considered waste to humans, decreasing the amount of pollution in
the world. As Terry Hazen, the head of the ecology department at the Lawrence Berkeley National, says, “There is
no compound, man-made or natural, that microorganisms cannot degrade” (Voigt, 2010). Furthermore,
microorganisms can complete different tasks depending on the type of microorganism, making them all unique.
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Escherichia Coli
Discovery. One of the most common bacterium known to man is Escherichia coli.
Theodor Escherichia first described it in 1885, and this “Gram-negative, rod-shaped bacterium”
was called Bacterium coli commune which was later changed to Escherichia coli, named after its
founder (Todar, 2008). For many years this microorganism was considered to be an organism
found in part of the large intestine. It was not until 1935, though, that a strain of Escherichia coli
was shown to be the cause of an outbreak of diarrhea among infants (2008). Escherichia coli was
found in all sorts of food, and whenever people handled infants infected with Escherichia coli,
they too obtained the bacterium causing it to spread rapidly (2008).
Uses of E.coli. In addition, Escherichia coli is the head of the large bacterial family
Enterobacteriaceae; the enteric bacteria are anaerobic Gram-negative rods that live in the
intestinal part of animals in health and disease (2008). The Enterobacteriaceae is one of the most
important bacteria medically because the bacteria in the family cause most of the leading causes
of death (2008). Escherichia coli are mostly harmless, but some strains can cause food poisoning
and other serious injuries. The dangerous thing about harmful Escherichia coli is that, once it is
established, an Escherichia coli strain may continue on for months or years (2008). If a strain of
Escherichia coli is hazardous, then it may be in one’s body for a long period of time, before
leaving, risking one’s life even more than if it was in one’s body for a short amount of time.
Escherichia coli can also respond to environmental signals such as chemicals, pH,
temperature, and many more, in a number of very remarkable ways because it is classified as a
unicellular organism (2008). For example, it can sense the presence or absence of chemicals and
gases in its environment and swim toward or away from them, or it can stop swimming and grow
fimbriae which will specifically attach the chemical to a cell, or surface receptor, which are also
known as protein molecules (2008).
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Overall, Escherichia coli are a well-known microorganism and the uses of this bacterium
are beneficial, as well as treacherous. This organism is easily grown and does not necessarily
need to stay inside the body because scientists have discovered that it is able to survive outside
the body for brief periods of time. Having said this, Escherichia coli can be found basically
anywhere including soil, water, food, and many more places (2008).
Temperature
Bioremediation. Temperature plays a key role in environmental problems, including
bioremediation. Margesin and Schinner (1997) said, “Most tests and experiments conducted by
scientists are carried out at temperatures higher than what is usually found in nature.” In general,
though, oil pollution is usually more extreme and serious in lower temperatures (1997). This
shows that there are more oil spills in lower temperatures where not a lot of bioremediation takes
place. According to Margesin and Schinner (1997), temperatures between 4oC and 30oC have an
insignificant number of microbial activities present during bioremediation.
For obtaining best results in bioremediation, higher temperatures and nutrients have to be
present (Braddock, Walworth, & Woodlard, 2001). These two factors also have to be properly
managed because extremely high temperatures can become harmful to some microorganisms,
lowering their bioremediation of oil (2001). This explains that many small factors in nature can
cause a huge impact on the environment, even something as little as changes in temperature.
Microorganisms. Microorganisms have been known to live in many extreme
environments including places where no photosynthesis occurs or where hot volcano lava is
present (Nguyen, 2006). In these different environments, microorganisms are able to adapt and
evolve over time so that they are able to live in extreme temperatures (2006). Warmer
Patel: Effect of Temperature on Bi
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temperatures cause many microorganisms grow faster and move more quickly. Colder
temperatures do the opposite, causing the bacteria to be in a slow-moving state (2006).
Escherichia coli. E.coli, as stated before is usually found on contaminated surfaces, like
food, water, and soil. Temperature can also affect how E.coli responds to an environment. A
study was conducted showing that E.coli grows best in room temperature rather than colder and
hotter climates (2006). E.coli is able to adapt to extreme temperatures, but this can take some
time to occur (2006). For example, if the temperature is very hot, E.coli is able to increase its
thermal optimum so that it can live in hotter climates. Colder temperatures, on the other hand,
usually freeze E.coli without killing the microorganisms. This causes E.coli to respond and move
slower than usual, which can lead to less biodegration of oil (2006).
Related Studies
Many related studies were done over the past years about bioremediation and the effect of
microorganisms on oil. For instance, a study took place in February 1996 in a remote, sandy
beach where scientists were studying the long-term effects of bioremediation agents on
biodegradation rate and the toxicity of oil (Delille et al., 2002). Ten experimental trials were
taken, and each trial received light Arabian crude oil and some even had bioremediation agents
added to them (2002). For example, one had fertilizer added, while another trial had fish
composts added (2002). This experiment went on for about three years, and during this time, not
many differences occurred, but high microbial populations were present during the experiment
(2002, pp. 118).
Another experiment about the biodegration of oil was done when many studies related to
bioremediation were tested to find the rate of biodegration of crude oil in seawater (Goldman et
al., 1993). These labs were done in a laboratory with and without nutrients and in a field with
Patel: Effect of Temperature on Bi
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and without nutrients (1993). In the results, data proved that if the oil concentration goes up, the
biodegration rate increases too (1993).
Another study took place after the Nakhodka oil spill accident in January 1997, when
microbial population changes were monitored immediately (Fujita et al., 2003). After the
experiment, it was concluded that oil-degrading activities in natural environments are regulated
by water temperature, dissolved oxygen, nutrient and salt concentrations, and much more (2003).
It was also proven that inorganic nutrients like ammonium, nitrate, and phosphate quicken the
process of oil spills’ cleanups (2003).
In 1997, a study was conducted where temperature was tested on oil biodegration by
yeast in soil (Margesin & Schinner, 1997). It was found that temperature affects the rates of
microbial activities because of the physical nature and composition of oil itself (1997). The
results of this experiment show that with a higher incubation time and with increasing
temperatures, the microorganism quantity present in the experiment also increased (1997). This
shows that with warmer temperatures, more microorganisms were present, causing the
biodegration to occur faster and more quickly.
At the University of North Wales, a study took place where microorganisms were added to
marine water with temperatures of 4oC and 14oC (Andrews, Gibbs, & Pugh, 1975). These waters
had a huge amount of nitrogen and phosphorous added. After the experiment, it was found that
the amount of nitrogen and phosphorous had not reduced greatly because of the cold
temperatures (1975). This experiment shows how colder temperatures cause microorganisms to
degrade wastes a lot slower than in normal, room temperature.
Conclusion
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Microbiology is an important subject to study, and it has helped technology improve
tremendously. Scientists know a plethora of ways microorganisms work after years and years of
investigating, researching, and experimenting. This research has also helped to demonstrate that
microorganisms can be used for environmental issues around the world and affect humans in
both positive and negative ways. For example, microbes can help cure and find illnesses for
disease, but they can also be detrimental, causing minor sicknesses, like diarrhea, and even major
illnesses, which later can cause death. Also, microorganisms can help the environment by
removing unwanted wastes made by humans. As stated before, this research shows that
microorganisms can eliminate oil and other hazardous substances from the environment more
quickly in warmer temperatures than in colder temperatures. Looking at the whole picture, the
study of microorganisms will continue to improve the world because knowledge is unlimited,
and scientists will continue figuring out more and more information about the world of microbes
and their reactions and effects to different temperatures.
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Materials

permanent marker

label tape

8 culture tubes with caps

3 pipettes (with graduation marks)

16 mL of nutrient broth

120 mm, in height, of motor oil (Castrol Oil 5W-30)

8 mL of microbial suspension (E.coli)

16 mL of 0.2% tetrazolium

ruler that measures in mm

incubator (37oC)

refrigerator (13.7oC)

thermometer

camera (optional)
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Procedure
1. The 8 culture tubes were numbered 1, 2, 3, 4, 5, 6, 7, and 8.
2. A clean, plastic pipette was used to add 4 mL of nutrient broth to tubes 2, 4, 6, and 8.
The pipette was discarded when finished.
3. A ruler was used to add 15 mm, in height, of oil (Castrol Oil 5W-30) to all 8 tubes,
using another pipette.
4. A clean, plastic pipette was used to add 2 mL of microbial suspension (E.coli) to
tubes 1, 2, 5, and 6. The pipette was discarded when finished.
5. A clean, plastic pipette was used to add 4 mL of 0.2% tetrazolium to tubes 1, 3, 5, and
the pipette was discarded when finished.
6. The caps of the culture tubes were placed on firmly.
7. The culture tubes were shook in a slow, stirring motion.
8. The height of the oil was measured in all 8 culture tubes at the initial time period (0
hours) and the solution was observed inside the tube.
9. Tubes 1, 2, 3, and 4 were placed in the incubator (37oC).
10. Tubes 5, 6, 7, and 8 were placed in the refrigerator (13.7oC)
11. For 192 hours, the culture tubes were observed on what was happening inside and
how the oil looked. The height of the oil was measured inside each tube every 24
hours, as well.
12. Pictures were taken, if necessary.
13. Steps 1-12 are repeated for a multiple of trials.
Patel: EFFECT OF TEMPERATURE ON BIOREMEDIATION
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Results
Table 2.1
The Effect of Temperature on Bioremediation
Time (h)
96
120
0
24
48
72
144
168
192
15.0
15.0
14.7
14.7
14.7
14.3
14.3
14.3
14.0
Standard Deviation (mm)
Standard Error (mm)
0.0
0.0
0.0
0.0
0.6
0.3
0.6
0.3
0.6
0.3
0.6
0.3
0.6
0.3
0.6
0.3
1.0
0.6
Refrigerator (13.7oC) – E.coli and Oil
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
15.0
0.0
0.0
14.3
0.6
0.3
14.3
0.6
0.3
14.3
0.6
0.3
14.3
0.6
0.3
14.0
0.0
0.0
14.0
0.0
0.0
13.7
0.6
0.3
Refrigerator (13.7oC) – Tetrazolium and Oil
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
15.0
0.0
0.0
15.0
0.0
0.0
14.7
0.6
0.3
14.7
0.6
0.3
14.3
0.6
0.3
13.7
0.6
0.3
13.7
0.6
0.3
13.3
0.6
0.3
Refrigerator (13.7oC) – Oil
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
14.7
0.6
0.3
14.7
0.6
0.3
14.3
1.2
0.7
13.7
0.6
0.3
13.7
0.6
0.3
13.3
0.6
0.3
13.3
0.6
0.3
13.3
0.6
0.3
Incubator (37oC) – Oil, E.coli, and Tetrazolium
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
13.3
0.6
0.3
13.0
0.0
0.0
13.0
0.0
0.0
12.3
1.2
0.7
11.3
1.2
0.7
11.0
1.0
0.6
10.3
1.5
0.9
9.3
1.5
0.9
Incubator (37oC) – E.coli and Oil
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
14.3
0.6
0.3
14.0
1.0
0.6
13.7
0.6
0.3
13.0
0.0
0.0
13.0
0.0
0.0
13.0
0.0
0.0
12.3
0.6
0.3
11.0
0.0
0.0
Incubator (37oC) – Tetrazolium and Oil
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
14.7
0.6
0.3
14.0
0.0
0.0
14.0
0.0
0.0
14.0
0.0
0.0
13.7
0.6
0.3
13.0
0.0
0.0
12.7
0.6
0.3
12.7
0.6
0.3
Incubator (37oC) – Oil
Mean (mm)
Standard Deviation (mm)
Standard Error (mm)
15.0
0.0
0.0
15.0
0.0
0.0
15.0
0.0
0.0
15.0
0.0
0.0
15.0
0.0
0.0
14.7
0.6
0.3
13.7
0.6
0.3
13.0
0.0
0.0
13.0
0.0
0.0
Refrigerator (13.7oC) – Oil, E.coli, and
Tetrazolium
Mean (mm)
Table 2.1 is showing the mean, standard deviation, and standard error of the height of motor oil, in mm, in room temperature and at 13.7oC.
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
20
Table 2.2
Bioremediation in the Refrigerator (13.7oC) – Oil, E.coli, and Tetrazolium
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
15.0
15.0
15.0
15.0
0.0
0.0
48
15.0
14.0
15.0
14.7
0.6
0.3
72
15.0
14.0
15.0
14.7
0.6
0.3
96
15.0
14.0
15.0
14.7
0.6
0.3
120
14.0
14.0
15.0
14.3
0.6
0.3
144
14.0
14.0
15.0
14.3
0.6
0.3
168
14.0
14.0
15.0
14.3
0.6
0.3
192
14.0
13.0
15.0
14.0
1.0
0.6
Table 2.2 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains motor oil,
E.coli, and tetrazolium and was placed in the refrigerator.
Table 2.3
Bioremediation in the Refrigerator (13.7oC) – E.coli and Oil
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
15.0
15.0
15.0
15.0
0.0
0.0
48
14.0
15.0
14.0
14.3
0.6
0.3
72
14.0
15.0
14.0
14.3
0.6
0.3
96
14.0
15.0
14.0
14.3
0.6
0.3
120
14.0
15.0
14.0
14.3
0.6
0.3
144
14.0
14.0
14.0
14.0
0.0
0.0
168
14.0
14.0
14.0
14.0
0.0
0.0
192
14.0
13.0
14.0
13.7
0.6
0.3
Table 2.3 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that includes E.coli and
motor oil and was placed in the refrigerator.
Table 2.4
Bioremediation in the Refrigerator (13.7oC) – Tetrazolium and Oil
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
24
15.0
15.0
15.0
15.0
0.0
48
15.0
15.0
15.0
15.0
0.0
72
15.0
15.0
14.0
14.7
0.6
96
15.0
15.0
14.0
14.7
0.6
120
15.0
14.0
14.0
14.3
0.6
144
14.0
14.0
13.0
13.7
0.6
168
14.0
14.0
13.0
13.7
0.6
192
14.0
13.0
13.0
13.3
0.6
0.0
0.0
0.0
0.3
0.3
0.3
0.3
0.3
0.3
Table 2.4 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains tetrazolium
and motor oil and was placed in the refrigerator.
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
21
Table 2.5
Bioremediation in the Refrigerator (13.7oC) – Oil
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
15.0
14.0
15.0
14.7
0.6
0.3
48
15.0
14.0
15.0
14.7
0.6
0.3
72
15.0
13.0
15.0
14.3
1.2
0.7
96
14.0
13.0
14.0
13.7
0.6
0.3
120
14.0
13.0
14.0
13.7
0.6
0.3
144
13.0
13.0
14.0
13.3
0.6
0.3
168
13.0
13.0
14.0
13.3
0.6
0.3
192
13.0
13.0
14.0
13.3
0.6
0.3
Table 2.5 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains only motor
oil and was placed in the refrigerator.
Table 2.6
Bioremediation in the Incubator (37oC) – Oil, E.coli, and Tetrazolium
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
14.0
13.0
13.0
13.3
0.6
0.3
48
13.0
13.0
13.0
13.0
0.0
0.0
72
13.0
13.0
13.0
13.0
0.0
0.0
96
13.0
13.0
11.0
12.3
1.2
0.7
120
12.0
12.0
10.0
11.3
1.2
0.7
144
11.0
12.0
10.0
11.0
1.0
0.6
168
9.0
12.0
10.0
10.3
1.5
0.9
192
8.0
11.0
9.0
9.3
1.5
0.9
Table 2.6 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains motor oil,
E.coli, and tetrazolium and was placed in the incubator.
Table 2.7
Bioremediation in the Incubator (37oC) – E.coli and Oil
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
14.0
15.0
14.0
14.3
0.6
0.3
48
13.0
15.0
14.0
14.0
1.0
0.6
72
13.0
14.0
14.0
13.7
0.6
0.3
96
13.0
13.0
13.0
13.0
0.0
0.0
120
13.0
13.0
13.0
13.0
0.0
0.0
144
13.0
13.0
13.0
13.0
0.0
0.0
168
12.0
12.0
13.0
12.3
0.6
0.3
Table 2.7 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains motor oil and
E.coli and was placed in the incubator.
192
11.0
11.0
11.0
11.0
0.0
0.0
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
22
Table 2.8
Bioremediation in the Incubator (37oC) – Tetrazolium and Oil
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
15.0
15.0
14.0
14.7
0.6
0.3
48
14.0
14.0
14.0
14.0
0.0
0.0
72
14.0
14.0
14.0
14.0
0.0
0.0
96
14.0
14.0
14.0
14.0
0.0
0.0
120
13.0
14.0
14.0
13.7
0.6
0.3
144
13.0
13.0
13.0
13.0
0.0
0.0
168
12.0
13.0
13.0
12.7
0.6
0.3
192
12.0
13.0
13.0
12.7
0.6
0.3
Table 2.8 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains motor oil and
tetrazolium and was placed in the incubator.
Table 2.9
Bioremediation in the Incubator (37oC) – Oil
Time (h)
Trial 1 (mm)
Trial 2 (mm)
Trial 3 (mm)
Mean
Standard Deviation
Standard Error
0
15.0
15.0
15.0
15.0
0.0
0.0
24
15.0
15.0
15.0
15.0
0.0
0.0
48
15.0
15.0
15.0
15.0
0.0
0.0
72
15.0
15.0
15.0
15.0
0.0
0.0
96
15.0
15.0
15.0
15.0
0.0
0.0
120
14.0
15.0
15.0
14.7
0.6
0.3
144
14.0
13.0
14.0
13.7
0.6
0.3
168
13.0
13.0
13.0
13.0
0.0
0.0
Table 2.9 shows the height of motor oil, in mm, after 192 hours for three trials, and it also displays the
mean, standard deviation, and standard error of the three trials for the test tube that contains motor oil and
was placed in the incubator.
192
13.0
13.0
13.0
13.0
0.0
0.0
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
Figure 2.1
Figure 2.1 illustrates the mean height of oil, in mm, in different temperatures, for all eight test tubes.
Figure 2.2
23
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
24
Figure 2.2 displays the mean amount of oil in the four test tubes that were placed in the refrigerator at
13.7oC.
Figure 2.3
Figure 2.3 displays the mean amount of oil in the four test tubes that were placed in the incubator at
37oC.
Results Summary
Table 2.1 shows the mean, standard deviation, and standard error for all 8 test tubes, while
Tables 2.2-2.9 individually show the mean, standard deviation, and standard error for each test tube.
These tables also display all three trials. Figure 2.1 displays all 8 test tubes, showing the mean of all three
trials. Four test tubes were tested in the refrigerator and the other four test tubes were placed in the
incubator. In addition, two test tubes in the refrigerator included E.coli and another two test tubes in the
incubator contained E.coli. Figure 2.1 also displays the error bars for this experiment showing the
confidence in these results. Figure 2.2 shows the mean amount of oil for the four test tubes that were
placed in the refrigerator, while Figure 2.3 illustrates the mean amount of oil for the four test tubes placed
in the incubator.
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
25
Conclusion
The hypothesis that the amount of oil would decrease more if Escherichia coli were
added to the oil in room temperature was supported by the results in this experiment. In Figure
2.1, it is shown that the oil in the test tube with bacteria, tetrazolium, and oil, that was placed in
the incubator had decreased the most amount of oil. According to Table 2.6, the oil in this test
tube decreased from 15 mm to 9.3 mm, on average. Also, the test tube with E.coli and oil that
was placed in the incubator, decreased from 15 mm to 11 mm. The two test tubes that were
placed in a temperature of 13.7oC and included microbial solution decreased a lot less than the
test tubes that were placed in the incubator. According to Figure 2.1, all four test tubes that were
placed in the incubator decreased and degraded more oil than the test tubes that were placed in
the refrigerator. This shows that even without microbial suspension, warmer temperatures
degrade oil a lot faster than colder temperatures. Figure 2.1 also shows, though, that the two test
tubes that included microbial suspension degraded a lot more oil than the test tubes without
E.coli added.
Even though the hypothesis was supported by the data collected, some error did take
place. The standard error, though, was between 0.0-1.0, showing that not a lot of error had taken
place in this experiment. In Figure 2.1, the error bars, which represent the overall distribution of
the data, were overlapping, thus illustrating that the data point could have landed on any part of
the error bar. Taking this into consideration, this experiment contained many types of errors that
could have occurred. One reason for this error could be the fact that the instrument used was not
accurate enough to measure the amount of solution that was placed into the test tubes. For
example, using a pipet may not be exact because some liquid could have fallen out when it was
being transferred to the test tube, or the precise amount of liquid was not put into the test tubes.
For instance, instead of adding 2 mL of tetrazolium, only 1.8 mL of tetrazolium could have been
added. Another possible conclusion for having error during the experiment could have been that
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
26
when measuring the height of the oil in each test tube the only instruments used were the human
eye and a ruler that did not go further than measuring millimeters, so the measurement in the
experiment may not have been as accurate. Furthermore, every 24 hours, the height of the oil
was measured, but during one trial the test tubes were lifted up and the height was measured,
which created more error during the experiment. When the test tubes were lifted up and the
height of the oil was measured, the oil was not at the exact level, so one part was higher than the
other, creating another error. Some of the test tubes, though, did not have big error bars and they
were very small, showing that not a lot of error took place. Overall though, the experiment did
not have really large error bars, but some error did take place.
This project is a replica of what happens around the world every day. Oil is usually
dumped into the ocean by oil spills that are made by humans, and, as this continues, the land and
oceans on earth start becoming hazardous and harmful to animals. Sometimes, though, bacteria
are able to take in and degrade the oil, helping the environment little by little. Some parts of the
warmer temperatures. This experiment shows how E.coli can degrade oil in warm and cold
temperatures, but are there other bacteria that do the same? In the future, different types of
bacteria could be used to test if they all degrade oil just like E.coli. Another way of expanding
this project could be adding other substances that degrade oil, like dispersants, and fertilizer, to
figure out which substance degrades oil the most or the quickest. Right now, though, it can be
said that E.coli is able to take in oil faster in warmer temperatures, rather than in colder
temperatures, helping everything and everyone in general.
Patel: EFFECT OF TEMPERTURE ON BIOREMEDIATION
27
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