- Prior to beginning work on this assignment, read the Oil Spill Bioremediation investigation manual. This lab will enable you to simulate the bioremediation of a marine oil spill.
- The Process
- Take the required photos and complete all parts of the lab assignment (calculations, data tables, etc.).
- Use the Lab Worksheet as a resource to complete the Lab Report Template.
Transfer any answers and visual elements from the Lab Worksheet into the Lab Report Template. You will submit the Lab Report Template through Waypoint in the classroom.
- The Assignment
- Make sure to complete all of the following items before submission:
- Before you begin the assignment, read the Oil Spill Bioremediation investigation manual; you may also wish to review the video, SCI207 – The Scientific Method (Links to an external site.).
- Complete all activities using materials in your kit, augmented by additional materials that you will supply. Photograph the setup following these instructions:
- When taking lab photos, you need to include in each image a strip of paper with your name and the date clearly written on it.
Use the Lab Worksheet as a resource to complete the Lab Report Template.
Don't use plagiarized sources. Get Your Custom Essay on
Need lab done for SCI207 Our dependence upon the environment details, reading, worksheet, & template attached
Just from $13/Page
- Must use at least two credible sources outside of the textbook and lab manual.
Oil Spill Bioremediation
Investigation
Manual
ENVIRONMENTAL SCIENCE
Made ADA compliant by
NetCentric Technologies using
the CommonLook® software
Key
Personal protective
equipment
(PPE)
goggles gloves apron
follow
link to
video
photograph
results and
submit
stopwatch
required
warning corrosion flammable toxic environment health hazard
OIL SPILL BIOREMEDIATION
Overview
In this investigation, students will simulate the bioremediation of a
marine oil spill. Bioremediation is the use of living things to clean
up environmental pollution (in particular, microorganisms that
consume oil). Students will apply a suspension of oil-degrading
microbes to a small amount of oil and chemical indicator in a
culture tube. A change in the color of the chemical indicator signi-
fies a breakdown in the chemical structure of the oil.
Outcomes
• Describe the chemical nature of oil
• Explore the general process microbes use to break down oil
Time Requirements
Preparation …………………………………………………………..
3
0 minutes
Activity 1: Bioremediation of Oil …………………….. 45 minutes, then
10 minutes a day for 3 days
2 Carolina Distance Learning
Table of Contents
2 Overview
2 Outcomes
2 Time Requirements
3 Background
7 Materials
8 Safety
8
Preparation
9 Activity
1
11 Submission
11 Disposal and Cleanup
12 Lab Worksheet
Background
Each year, millions of gallons of oil enter the
world’s oceans. The impact of oil pollution on
marine ecosystems is profound and long-lasting.
Dramatic accidents and oil spills—like the 2010
Deepwater Horizon disaster (also known as
the BP oil disaster)—make headline news. The
oil that spilled during the Deepwater Horizon
disaster, for example, decimated bird and
fish populations and resulted in the deaths of
dolphins, turtles, and deepwater corals. It also
negatively impacted the commercial fisheries in
the Gulf of Mexico. Likewise, the harmful effects
of the 1989 Exxon Valdez oil spill are still felt
in Alaska’s Prince William Sound, more than a
quarter century after the tanker ran aground and
spilled more than 10 million gallons of crude oil.
However, these large incidents account for
only about 5% of the oil polluting the seas that
is a result of human activity. The vast majority
of ocean oil contamination originates from
the accumulation of smaller, less publicized
but commonplace events, such as leaks from
smaller oil tankers, routine operation of oceanic
oil wells, leaking storage tanks and pipelines for
offshore oil wells, and improperly drilled holes in
the ocean floor. Loading and unloading tankers
with oil for transfer from offshore rigs to onshore
sites also can introduce oil into the ocean.
Refined oil (i.e., fuel oil, gasoline, and other
processed petroleum products) from municipal
and industrial sources is often accidentally or
deliberately dumped, spilled, or leaked on land
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3
Figure 1.
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OIL SPILL BIOREMEDIATION
Background continued
and into waterways. Oil on roadways from motor
vehicles is carried to waterways and eventually
to the ocean by heavy rainfalls. Many people
also dispose of used motor oil improperly. This
oil can enter storm drains, streams, and rivers
and can be carried out to sea. Oil can also enter
the atmosphere as smoke from oil fires and then
be deposited into the ocean and tributaries with
precipitation.
Crude oil is a complex mixture of several types
of hydrocarbons. A hydrocarbon is a chem-
ical compound made entirely of carbon and
hydrogen that usually forms a long chain, which
can be either linear or in the form of a ring
(see Figure 2). The toxic chemicals floating in
oil can kill or contaminate plankton and algae.
When fish eat these contaminated foods,
they can also become contaminated or even
die. Fish larvae can be killed, sickened, or
disfigured, negatively impacting future popu-
lation numbers. The larvae that survive likely
continue to consume oil as well. These
compounds are often transferred through an
entire food web and can become more concen-
trated (have a higher potency) when larger fish,
birds, other animals, and humans prey on these
contaminated fish. This process is referred to
as bioaccumulation. Heavy oil components
sink to the ocean floor where they cover benthic
(bottom-dwelling) organisms, such as crabs,
oysters, mussels, and clams. The toxicity of
the oil either kills these organisms or pene-
trates their tissues, making them dangerous to
consume. Oil also coats the feathers of birds
and the fur of marine mammals, causing them
to lose their natural insulation, buoyancy, and
motility. Many of these animals drown; others
die due to loss of body heat.
Natural seepage of oil from oceanic oil deposits
accounts for a significant amount of oil released
into the ocean, but much of this natural seepage
is consumed by ocean-dwelling bacteria that
have evolved specialized pathways that enable
them to use oil as food and convert it into
energy. These microbes, containing mostly
bacteria and some fungi, break down the long
chain hydrocarbons of petroleum and chem-
ically convert them into energy and nutrients
for their own biological processes, which is
known as biodegradation. The hydrocar-
bons act as a carbon source from which the
organisms build their biomass and grow. Many
different species of oil-degrading microbes work
together to break down the components of oil.
These marine bacteria and fungi use enzymes
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4 Carolina Distance Learning
Figure 2.
Heptane
Chain
Benzene
Ring
and oxygen in seawater to break down the
ring structures of the hydrocarbons, producing
carbon dioxide (CO2) in the process.
Scientists recognize great potential in utilizing
oil-degrading microbes to expedite the break-
down of harmful oil from spills. There are three
major approaches to this process, known as
bioremediation, to clean up marine oil spills:
1. Manipulate the nutritional composition of the
spill site to enhance the activity of indigenous
oil-eating microbes.
2. Augment naturally occurring microbes with
special mixtures of non-native, oil-degrading
microorganisms.
3. Utilize genetically engineered microorganisms
specifically designed to degrade oil
effectively.
Optimizing the environment of oil-degrading
microbes to accelerate their growth and repro-
duction is called biostimulation. Nutrient
availability is the rate-limiting factor in micro-
organisms’ ability to degrade petroleum in
bioremediation. Nutrients such as nitrogen,
phosphorous, and iron are necessary for indig-
enous oil-degrading microbes to convert the
petroleum hydrocarbons into useful biomass
and nontoxic by-products. These nutrients are
typically in short supply because non-oil-
degrading microorganisms compete to consume
them. Through biostimulation, nutrients are
added to the oceanic environment, much like
applying fertilizer to a lawn. The amplified
nutrient supply increases the rate and extent of
microbial oil degradation. However, in order to
encourage maximum microbe growth and oil
breakdown, the nutrients must remain in contact
with the oil and their concentration must remain
at an optimal level for an extended time period.
These conditions are difficult to maintain in
dynamic aquatic systems.
A variety of chemical methods have been
employed to disperse (break oil into smaller
droplets) oil spills. The rationale for using disper-
sants is that, by breaking down the oil slick (area
of oil floating on a body of water), the surface
area of the oil is increased to allow more of the
oil to be available for degradation by microbes.
Chemical dispersants, also called surfactants,
are classed by their ionic charge. Soaps are
an example of an anionic (negatively charged)
surfactant. The use of chemical dispersants
is controversial, both because increasing the
surface area of the oil also makes the toxins
within the oil more available to the environment
and because of the toxic nature of many of the
surfactants.
The process of supplementing or “seeding” a
population of naturally occurring, oil-degrading
microbes with additional microorganisms is
called bioaugmentation. This technique is often
used when the existing population of microbes
in a contaminated region is not optimally suited
to degrade the type of oil present. Mixtures of
microbial species that are better decomposers
can be combined and grown in large batches
in laboratories and then introduced at the spill
site in bulk. Oil-degrading microbes can easily
be cultivated in large quantities in laboratories
and be ready for use when ocean oil pollution
occurs. These microbial populations, called
seed cultures, can be stored for up to three
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OIL SPILL BIOREMEDIATION
Background continued
years. However, recent studies following the
cleanup effort in the wake of the Deepwater
Horizon oil spill have questioned the efficacy of
seeding microbe populations.
Breaking down petroleum is a complex biolog-
ical process. A single species of an oil-
degrading microbe cannot achieve complete
degradation on its own. However, scientists are
creating genetically engineered microorganisms
(GEMs) in hopes to accomplish what natural
species cannot. These GEMs are designed to
incorporate the pathways and enzymes neces-
sary to degrade oil more efficiently and thor-
oughly. The use of GEMs at spill sites has great
potential, but it is a relatively new approach that
is undergoing continued development.
In this investigation, you will simulate the biore-
mediation of a marine oil spill using microorgan-
isms that consume oil. Rid-X® is a mixture of
bacteria and enzymes that is used to maintain
septic systems by degrading sewage, including
oils. A chemical called tetrazolium is used as an
indicator for the breakdown of oil. Tetrazolium
typically is colorless (when oxidized) but turns
pink when its chemical composition is changed
(when reduced). When microorganisms break
down the carbon compounds in oil, they create
by-products that serve as electron donors
(reducing agents). These electron donors change
the chemical composition of the tetrazolium
indicator (by the addition of hydrogen), causing
it to turn pink (due to the creation of an insoluble
pink compound). In this activity, the reduction of
tetrazolium from its oxidized, colorless form to
its reduced, pink form is used as an indication
that the breakdown of oil is taking place.
The use of tetrazolium in this activity marks
only the beginning of oil degradation. Complete
decomposition of the oil would require abundant
nutrients, involve several species of microbes,
and occur over a long period, which cannot be
completed in a scaled-down setting such as
the one being carried out in this experiment.
However, basic changes in oil composition can
still be observed.
6 Carolina Distance Learning
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Materials
Included in the materials kit:
10 plastic
pipets, 3 mL,
graduated
6 Culture
tubes with
caps
Rid-X® Septic
System
Treatment
(powder),
20 g
Funnel
2 Plastic cups
Cheesecloth
Test tube rack
Bag
containing
tetrazolium
indicator
powder
(0.02 g) and
distilled water
(100 mL),
0.02%
Graduated
cylinder
Needed from the equipment kit:
Reorder Information: Replacement supplies
for the Oil Spill Bioremediation investiga-
tion can be ordered from Carolina Biological
Supply Company, item number 580830.
Call: 800.334.5551 to order.
Needed but not supplied:
• Cooking oil
• Bottled water, 30 mL
• Warm tap water, 140 mL
• 10% bleach solution for cleanup
• Pencil
• Stopwatch (or cell phone with a timer)
• Camera (or cell phone capable of taking
photographs)
Preparation
Many factors can affect the speed with which
microbes break down substrate (i.e., tempera-
ture, salinity, availability of limiting nutrients,
exposure to sunlight, and access to the
substrate). In this activity, you will design an
experiment to test the effects of one of these
factors on the breakdown of oil using materials
from your environment. Two tubes are provided
in your kit to perform your test.
1. Read through the activities.
2. Obtain all materials.
3. Begin preparing the microbial suspension as
follows:
a. Using the graduated cylinder, measure out
140 mL of warm tap water in a plastic cup.
b. Add the powdered contents of the Rid-X®
Septic System Treatment container (~20 g)
to the cup, and mix thoroughly by gently
swirling the cup.
c. Let the mixture sit undisturbed for
about 15 minutes to allow undissolved
matter to settle.
After the undissolved matter has settled in
the cup, continue preparing the microbial
suspension, as follows:
d. Fold the cheesecloth in half to double it.
Place the folded cheesecloth in a funnel in
the top of a second cup.
e. Slowly pour the Rid-X® mixture into the
cheesecloth in the funnel to filter the pulp
from the microbial suspension.
f. Dispose of the cheesecloth and pulp.
OIL SPILL BIOREMEDIATION
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8 Carolina Distance Learning
Safety
Wear safety goggles,
gloves, and a lab apron
at all times while conducting this investigation.
Read all the instructions for this laboratory
activity before beginning. Follow the instructions
closely, and observe established laboratory
safety practices, including the use of appropriate
personal protective equipment (PPE).
Tetrazolium may cause skin and eye
irritation. Rid-X® Septic System Treat-
ment contains bacterial spores and
enzymes and may cause lung irritation. Avoid
contact with skin and eyes. Avoid breathing dust
particles.
Household bleach can damage eyes
and skin, and it should not be ingested.
It should be used only in a well-
ventilated area, such as a room with an
open window or a bathroom with a
ventilation fan. Always wear PPE
(goggles, an apron, and gloves) when handling
bleach. If bleach gets into your eyes, rinse them
thoroughly with water for several minutes. If
you are wearing contact lenses, remove them if
practical to do so and then continue rinsing. If
bleach contacts your skin, wash thoroughly with
soap and water. If bleach contacts your clothing,
remove and wash it before wearing it again. If
ingested, do not induce vomiting. Rinse your
mouth thoroughly with water, and seek medical
attention immediately.
Do not eat, drink, or chew gum while performing
this investigation. Wash your hands with soap
and water before and after the investigation, and
sanitize the work space with a 10% bleach solu-
tion after finishing. Keep pets and children away
from lab materials and equipment.
This will require 3 days of data collection
once the experiment is set up. Please
plan accordingly.
Bioremediation of Oil
1. Use a pencil to label the culture tubes 1
through 6.
2. Tubes 1 and 2 will utilize the tetrazolium
indicator to determine if metabolism is taking
place. Hypothesize whether you think the
indicator will change color in both tubes,
neither one, or just tube 1 or tube 2, and
describe your reasoning. Use Figure 3 to help
form your hypothesis. Record this information
in the “Hypotheses” section in your Lab
Worksheet.
3. Tubes 3 and 4 will be used to examine
the change in appearance and physical
properties of the oil in the presence of
microorganisms. Hypothesize if you will be
able to see the oil broken down in either of
these tubes without the indicator present, and
describe your reasoning. Use Figure 3 to help
form your hypothesis. Record this information
in the “Hypotheses” section in your Lab
Worksheet.
4. Tubes 5 and 6 will be used to test the effects
of your chosen environmental factor on
the breakdown of oil by microorganisms.
Choose one of the following as your
environmental factor to alter (please check
with your instructor if you would like to use
a different option): amount of microbes
present, amount of oil present, type of oil
present, light conditions, or temperature
variation. Hypothesize which of the two tubes
will experience the greater breakdown by
microbes, and describe your reasoning. Use
Figure 3 to help form your hypothesis. Record
this information in the “Hypotheses” section
in your Lab Worksheet.
ACTIVITY
ACTIVITY 14. To prepare the tetrazolium solution, pour the
100 mL of distilled water from the enclosed
bottle into the enclosed brown bottle
containing the tetrazolium powder. Be sure
to do this slowly. Shake the brown bottle
well to ensure the powder dissolves into
solution.
continued on next page
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ACTIVITY
ACTIVITY 1 continued
5. Using a plastic pipet, add 1 mL of
0.02% tetrazolium indicator (made in the
“Preparation” section) to tubes 1, 2, 5, and
6 only. Use the graduations marked on
the plastic pipet to measure 1 mL. Discard
the pipet when finished. (See Figure 3 for
guidance in adding quantities.)
6. Using a clean plastic pipet, add 2 mL of
distilled water to tubes 1, 2, 3, and 4 only.
Use the graduations marked on the plastic
pipet to measure 2 mL. Discard the pipet
when finished.
7. Using a clean plastic pipet, add 10 drops of
oil to all six tubes. Discard the pipet when
finished.
8. Using a clean plastic pipet, add an additional
2 mL of distilled water to tube 1, an additional
3 mL to tube 3, and an additional 1 mL to
tube 4. Use the graduations marked on
the plastic pipet to measure each amount.
Discard the pipet when finished.
9. Using a clean plastic pipet, add 2 mL of
microbial suspension to tubes 2, 4, 5, and
6 only. Use the graduations marked on the
plastic pipet to measure 2 mL. Discard the
pipet when finished.
10. Add the required component(s) or set up
the required changes for your experimental
design to tubes 5 and 6, and record what
you did in Data Table 3 of the “Observations/
Data Tables” section of the Lab Worksheet.
11. Cap all culture tubes. Mix the liquid in all six
tubes by finger vortexing them, one tube at
a time. To finger vortex, hold the top of the
tube securely in one hand; draw the index
finger of the other hand toward you several
times, gently tapping the side of the tube
near the bottom. This creates a whirlpool
inside the tube, which mixes the liquids.
Repeat this procedure with the remaining
tubes. Place all six tubes upright in a test
tube rack.
Finger Vortexing
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12. Record observations about the color,
viscosity, and general appearance
of the oil, today and for the following three
days, in the corresponding data tables in the
Lab Worksheet. Also, take photographs all
four days. The photographs should show all
6 tubes (with their labels clearly visible) as
well as a strip of paper with your name and
the date written on it. You will be uploading
these photographs to your lab report. Use
the following tips to help clearly distinguish
the differences between the tubes:
a. To aid in your observations, finger vortex
the tubes daily, as described in Step 11.
b. Manipulate the tubes in any way
that allows you to better view the
characteristics of the oil in each.
c. In particular, to aid in your observations of
tubes 3 and 4, hold them up to the light in
a horizontal position and observe how the
oil moves over the liquid.
d. Invert tubes 3 and 4 several times and
watch the oil gather back at the top of
the liquid. Observe any differences in the
composition of the oil.
13. Gently loosen each cap on each
culture tube by turning it 45 degrees
counterclockwise. Let the tubes sit
overnight.
Just before observing the tubes each day,
retighten the caps on the tubes. After
observing the tubes, loosen the caps again
as explained in Step 13.
Submission
Using the Lab Report Template provided,
submit your completed report to Waypoint for
grading. It is not necessary to turn in the Lab
Worksheet.
Disposal and Cleanup
1. Dispose of solutions down the drain with the
water running. Allow the faucet to run a few
minutes to dilute the solutions.
2. Rinse and dry the lab equipment from the
equipment kit, and return the materials to
your equipment kit.
3. Dispose of any materials from the materials
kit in the household trash. The plastic funnel
may be recyclable.
4. Sanitize the work space with a 10% bleach
solution, and wash your hands thoroughly.
Disinfecting a Surface
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ACTIVITY
Lab Worksheet
continued on next page
12 Carolina Distance Learning
Hypotheses
Activity 1.
Tubes 1 and 2:
Tubes 3 and 4:
Tubes 5 and 6:
www.carolina.com/distancelearning 13
Observations/Data Tables
Data Table 1.
Data Table 2.
Tubes 1 and 2 Observations
Day Tube 1 Tube
2
0
(Initial setup)
1
2
3
Tubes 3 and 4 Observations
Day Tube 3 Tube 4
0
(Initial setup)
1
2
3
ACTIVITY
14 Carolina Distance Learning14 Carolina Distance Learning
Data Table 3.
Tubes 5 and 6 Observations
Chosen environmental factor to change:
Day Tube 5 Tube 6
0
(Initial setup)
1
2
3
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NOTES
http://www.carolina.com/distancelearning
ENVIRONMENTAL SCIENCE
Oil Spill Bioremediation
Investigation Manual
www.carolina.com/distancelearning
866.332.4478
Carolina Biological Supply Company
www.carolina.com • 800.334.5551
©2019 Carolina Biological Supply Company
CB781651908 ASH_V2.2
http://www.carolina.com/distancelearning
http://www.carolina.com
Runninghead: NAME OF LAB
1
Running head: NAME OF LAB
3
Name of Lab
Your Name
SCI 207: Our Dependence Upon the Environment
Instructor’s Name
Date
*This template will enable you to produce a polished Lab Report. Simply complete each section below, pasting in all your completed data tables, graphs, and photographs where indicated. Before you submit your Lab Report, it is recommended that you run it through Turnitin, using the student folder, to ensure protection from accidental plagiarism. Please delete this purple text, and all the instructions below, before submitting your final report.
Title of Lab Goes Here
Introduction
Background paragraph: Provide background on the lab topic, explaining the key concepts covered in the lab and defining (in your own words) important terms relating to the lab. Explain why the lab topic is important to scientists. Using
APA format
, cite at least two outside credible sources (sources other than textbook or lab manual) in your statement.
Your background paragraph should be 5-7 original, substantive sentences long.
Objectives paragraph:
In 4-5 sentences, explain the purpose of this lab. What is it intended to examine or test?
Hypotheses paragraph: State your hypotheses for this lab. Be sure to cover all the lab activities, one at a time. For each hypothesis, explain why you originally thought that would happen.
Note: Do not mention the actual results of the lab here – they go later in the report.
For additional help in writing your Introduction section, refer to the Ashford Writing Center Resource,
Introductions and Conclusions
.
Materials and Methods
Using your own words, describe what you did in each of the lab activities. Answers should enable a lab report reader to repeat the lab just as you did it – a process known as replication. Clearly explain any measurements you made (including the measurement units).
Results
Data Tables: Copy and paste each of your completed data tables here, in order (Weeks One, Two, Four, and Five Labs only).
Observations: Provide your observations for each lab activity here, in order (Week Three Lab only)
Graphs: Paste your graphs here (Week Four Lab only). Include a numbered figure caption below each one, in APA format.
Photographs: Paste your photographs here, in the order they were taken in the lab. Include numbered figure captions below each one, in APA format.
For additional help with the data tables and images, refer to the Ashford Writing Center resource,
Tables, Images, and Appendices
.
Discussion
Accept or reject hypotheses paragraph: Based upon the results of each lab activity, explain whether you accepted or rejected each of your hypotheses, and why.
Follow these steps:
· Restate your original hypothesis for the lab activity.
· Communicate the results of the lab. Then,
· Compare your hypothesis to the results of the lab and decide whether to accept your hypothesis or reject it.
· State if your hypothesis is supported or not, and explain with evidence.
· Move on to the next lab activity and repeat the process.
What I have learned paragraph: What important new things have you learned from this lab? Use at least one credible outside source (not the lab manual or textbook) to answer this question. Cite the source using APA format.
Answers should be 5-7 original, substantive sentences in length.
Sources of error paragraph: What challenges did you encounter when completing this lab? (Identify at least one.) How might those challenges that you experienced have affected the accuracy of the results that you obtained?
Future research paragraph: Based upon what you learned in this lab, what new questions do you have about the topic of this lab? In a few sentences, how might you design a new lab activity to answer those questions?
References
List the references that you cited in your report, in APA format and alphabetically by author’s last name.
If you did not actually cite the source somewhere in your paper, do not include it.
For additional help in formatting your resources section, refer to the Ashford Writing Center’s resource for
Formatting your Reference List
.
Lab Worksheet
Hypotheses
Activity
1
.
Tubes 1 and
2
:
2 Carolina Distance Learning
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3
Tubes 3 and 4:
Tubes 5 and 6:
Observations/Data Tables
Data Table 1. Tubes 1 and 2 Observations
|
Day
|
Tube 1
|
Tube 2
|
|
0
(Initial setup)
|
|
1
|
|
2
|
|
3
|
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Data Table 2. Tubes 3 and 4 Observations
Day
Tube 3
Tube 4
0
(Initial setup)
1
2
3
Data Table 3. Tubes 5 and 6 Observations
Chosen environmental factor to change:
Day
Tube 5
Tube 6
0
(Initial setup)
1
2
3
