1

Coen Smith
01-18-2023
I.S.E.F. 2023
Polyethylene Reared Mealworms as Human Sustenance

Rationale:
According to the World Food Programme, 828 million people worldwide are
malnourished, with 6% of those, originating from forty-nine countries, being close to famine.1 If
the harrowing state of the human population is not enough to cause worry, surely the statistics
provided by Earth.Org can elicit a reaction; Approximately 300 million tons of plastic are
produced each year worldwide, and roughly sixty percent of said plastic ends up scattered in
landfills and the environment,2 where it will take over 700 years to even begin breaking down.3
In this experiment, I plan to test a simple solution. According to a 2019 study by Stanford,
mealworms can consume multiple varieties of plastic and even possess the ability to excrete the
toxic chemical hexabromocyclododecane present in certain plastics.4 This capability means that
they are able to be reinstituted into the food chain without any negative effects on higher
organisms. Presently, there are no studies on the protein content of mealworms raised on any
type of plastic. Therefore, I propose that we study the effects of polyethylene fodder on the
protein content of mealworms in order to determine if said mealworms would be a viable source
of protein for human consumption if fed plastic that would otherwise pollute our planet.

Experiment and Line of Reasoning:

For the experiment, we will compare the protein content of thirty-six mealworms raised
on polyethylene and the protein content of thirty-six mealworms raised on brown rice. The
number thirty-six was chosen as a sample size as we can then raise twelve groups apiece, with
five in each group, and then randomly select three from each group for testing. This allows us
room for error if a few die. This also is a large enough sample size where we can statistically
prove that our results are accurate. Polyethylene was chosen as it is the most widely produced
plastic, and therefore there is a large enough supply for this experiment to have an impact in the
future. Brown rice was chosen as a comparison because brown rice is the most widely grown and
consumed grain in the countries affected by the issues this experiment is attempting to address.

1
“As many as 828 million people go to bed hungry every night… A total of 49 million people in 49 countries are teetering on the edge of
famine.” This came from https://www.wfp.org/global-hunger-crisis.
2
“Our reckless plastic use and consumption has driven the world to generate approximately 300 million tonnes of plastic waste each year to keep
up with demand, 60% of which ends up in our natural environment or landfills.” This was sourced from
https://earth.org/plastic-pollution-statistics.

3
“Under normal conditions in nature, plastic bottles (usually made of polyethylene, also known as PET, or polyethylene terephthalate) will begin
to break down only after 500-700 years and even then, the process will be very slow.” This was found at
https://davidson.weizmann.ac.il/en/online/askexpert/why-does-it-take-plastic-so-long-break-down.
4
“Mealworms fed a steady diet of HBCD-laden polystyrene were as healthy as those eating a normal diet. The same was true of shrimp fed a
steady diet of the HBCD-ingesting mealworms and their counterparts on a normal diet.” This came from
https://news.stanford.edu/2019/12/19/mealworms-provide-plastic-solution.
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Brown rice was chosen over white rice because it is most likely what the poverty stricken will be
consuming as it is easier to process and has more nutritional value.

Hypotheses:
The independent variables are the polyethylene pellets and the brown rice. The dependent
variables are the protein content in the mealworms after ten days of testing. Some of the many
control variables are the one quart mason jars used as habitats,5 the feeding schedule,6 and the
testing process.7 Therefore, if mealworms are raised on polyethylene, then the mealworms will
not have a high protein content, because polyethylene does not have a high nutritional value
which will not stimulate growth. Additionally, if mealworms are raised on brown rice, then the
mealworms will have a high protein content because brown rice has a high nutritional value
which will stimulate growth.

Procedure Outline:
The worms will be kept in a total of twenty-four jars. Each jar will contain five
mealworms. Twelve of the jars will be raised on polyethylene, and the other twelve will be raised
on brown rice. Each day, the worms' food will be replenished, and they will be misted twice
daily. After ten days, the worms will be removed from their habitat and humanely dispatched.
The worms will then be dehydrated, and using a BCA assay kit and a spectrophotometer we will
determine the worm's protein content by comparing the results against a standard curve.

Mealworm Living Conditions:

The mealworms will first be acquired at PetSmart. They will be The Bug Company, and
will come in a pack of 500.8 The worms will be immediately transported to their habitat. As we
only need 120 of them, the remaining 380 will be released into the wild upon purchase. The
worms will be housed in one quart Ball mason jars. The jars will be fully sanitized with
isopropyl alcohol, left to dry very briefly, and then filled with 50mg each of either polyethylene
or uncooked brown rice. Five worms will be added to each jar, and then the worms will be
misted until they are all damp. Coffee filters will be placed over the top of each jar, and secured
with a rubber band. Each day, the worms will be misted twice, and they will be fed once. The
misting will take place at either 6:00 am and 8:00 pm or 8:05 am and 3:45 pm depending on
where the worms are kept. The worms will be fed at the latter of the misting sessions. This
process will repeat for ten days.

Mealworm Processing and Justification:

5
These jars can be found at
https://www.ballmasonjars.com/products/jars/canning-jars/ball%C2%AE-mason-jars-lids%2C-regular-mouth/SAP_62000.html.
6
See page two.
7
See page four.
8
These worms can be found at https://www.petsmart.com/reptile/food/the-bug-company-mealworms-5230294.html.
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At the conclusion of ten days, the testing will take place. Three worms will be removed
from each of the twenty-four containers, and the remaining two worms from each container will
be released into the wild. The worms to be tested will be isolated from each other in labeled
groups based on which of the two they were raised on, and they will be frozen.9 As soon as it is
determined that the worms are no longer active, they will be removed from the freezer and they
will be moved to the dehydrator.10 After one cycle in the dehydrator, which will last a couple of
hours, the dehydrated worms will be powdered and stored separately but labeled until they are
ready to be tested on.

The use of mealworms cannot be avoided. The experiment is based fully on the use of
mealworms as an alternative protein, and for that to be the case some will need to perish. For this
experiment, we are using the fewest number that will lend a statistically significant and accurate
assay. Additionally, in our purchase of 500, we are saving 380 of them from a certain death of
being eaten by a reptile. Ignoring the above, fact of the matter is, we must be willing to accept
that in order for us to eliminate polyethylene in the environment, and for us to better the 848
million malnourished people on this Earth something has to be sacrificed, and in this case we are
just feeding two diners with one scone.

Detailed Procedure11:
Materials:
➢ Twenty-four Ball Mason Quart Jars
➢ Isopropyl Alcohol
➢ Paper Towels
➢ VViViD Eco Friendly High Density Polyethylene Plastic Packing and Polyfill Pellets
➢ Scale
➢ The Bug Company Mealworms, 500 count
➢ Misting Apparatus
➢ Distilled Water
➢ Unbleached Coffee Filters
➢ Rubber Bands
➢ Twenty-four Mini Ball Mason Jars and Screw Lids
➢ Freezer
➢ Dehydrator
➢ Mini Coffee Grinder
➢ Pierce Rapid Gold BCA Protein Assay Kit

9
Freezing insects has been known to make them less active, and to numb pain. Doing this will make sure that they do not feel as much discomfort
as they are being dispatched in the dehydrator.
10
Dehydrating the mealworms is important for two reasons. Firstly, the mealworms will most likely be consumed dehydrated or roasted, so the
water will effectively be eliminated. Including the natural water in our experiment could throw off the results. Second, the worms cannot be
reduced to a powder if they are moist.
11
May change as needed, and materials will be updated to be more detailed as they are acquired.
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➢ Eighty Cuvettes
➢ .1 ml - 10 ml Pipette
➢ 10 µl - 100 µl Pipette
➢ Test Tubes and Stoppers
➢ Graduated Cylinder
➢ 1 N HCl12
➢ Spectrophotometer with 480 nm filter

Procedure:

Setting Up and Caring for the Mealworms:

1. Take twenty-four Ball Mason One Quart Jars and rinse with distilled water. Swab the
inside with isopropyl alcohol on a paper towel. Let them dry. Label each jar in some
fashion.
2. While the jars are drying, acquire the mealworms. Keep the mealworms in the fridge until
use.
3. Using the scale, weigh out 250mg of polyethylene pellets and place them in each of the
jars. Mist well with distilled water.
4. Add five mealworms to each jar, mist them thoroughly, and cover the jars with
unbleached coffee filters. Use a rubber band to secure the coffee filters so that they cover
the entire mouth of the jar.
5. The remaining mealworms should be brought outside and distributed among an area with
a lot of vegetation, and should not be placed near a beehive.
6. For the next ten days, repeat the following process: At 6:00 am or 8:05 am mist the
worms with distilled water. At 8:00 pm or 3:45 pm mist the worms again, and also add
250 mg of polyethylene pellets to each jar. Keep detailed notes and refer to the labels on
the jars.

Preparing Worms for Testing:

1. Label each of the mini mason jar lids by jar. Ex: The first lid has been sharpied, “Poly
One”, to represent that it was the first quart jar and contained the polyethylene worms.
2. Take twenty-four mini Ball Mason Jars and rinse with distilled water. Swab the inside
with isopropyl alcohol on a paper towel. Let them dry.
3. Select three worms at random from each quart jar, and place in a mini jar. As soon as you
do so, place the corresponding mini lid on the top of the jar.
4. Transport all of the mini jars to the freezer, and leave them there for one hour.

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A specific concentration of hydrochloric acid.
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5. After one hour, remove the jars and tip them over so all of the mealworms are resting on
the bottom of the lids. Unscrew the jars so that the lids are flat on the table with the
mealworms inside.
6. Transport all of the jar lids onto a sheet tray and place in the dehydrator at sixty degrees
celsius. Let dehydrate for three to four hours, or until dry.
7. While all of the worms are dehydrating, take the twenty-four mini Ball Mason Jars and
rinse with distilled water. Swab the inside with isopropyl alcohol on a paper towel. Let
them dry.
8. When the worms are dehydrated, transfer to the jars and leave the lids slightly ajar to
allow airflow.
9. Take the test tubes and rinse with distilled water and isopropyl alcohol. Let them dry, and
then label them to match the mini Ball Mason Jars. Weigh the tubes and label
accordingly.
10. Take the mini coffee grinder, and swab the inside with isopropyl alcohol on a paper
towel. Let it dry. Add one worm, and grind. Place in a test tube that corresponds to the
label on the jar. Note: You will have three samples for each jar. Stop the test tube, and
repeat the process.

Preparing Diluted Albumin Standards:

1. Must ask Mrs. Phillips.

Preparing Working Reagent:

1. We used the formula (# standards + # unknowns) × (# replicates) × (volume of WR per
sample) to find the amount of working reagent required. There are nine standards,
seventy-two unknowns, one replicate per each, and the volume of reagent per sample is
2 ml. This gives us (9 + 72) x (1) x (2 ml) = 158 ml. We also need a water blank, and we
would like room for error so we will make 204 ml.
2. Mix fifty parts of Rapid Gold BCA Reagent A with one part Rapid Gold BCA Reagent B
(50:1, Reagent A:B). This means we need 200 ml of Reagent A and 4 ml of Reagent B.

Preparing For Final Test and Testing:

1. Prepare eighty cuvettes for testing.13 The specific process depends on what material your
cuvette is made of.
2. On Google Sheets or Excel prepare a grid that shows exactly where all the cuvettes are
based on what they will contain.
3. Measure each test tube along with the sample, and subtract the weight you previously
labeled on the tube. Add an equivalent weight of distilled water and mix well.
4. Pipette 80µl of each standard, unknown, and one distilled water blank into each
corresponding cuvette.

13
This is seventy–two unknowns, nine standards, and one water blank.
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5. Add 2 ml of the working reagent to each of the cuvettes and mix well.
6. Incubate tubes for five minutes at room temperature.
7. Add 500µl of 1 N HCI.
8. Set the spectrophotometer to 480 nm, and insert the water blank. Zero it out.
9. Measure the absorbance of all the samples within thirty minutes.
10. Record the absorbance, and then subtract the absorbance of the water blank from the
absorbance of all the other samples, including the standards.
11. Prepare a standard curve by plotting the average blank-corrected 480 nm measurement
for each BSA standard versus its concentration in µg/mL. Use the standard curve to
determine the protein concentration of each unknown sample.

Disposal of Mealworms:
All the mealworms that have been exposed to any chemicals will have been used up by
the end of the experiment. All the remaining, living mealworms will be completely free of any
chemicals and will be released into the wild.

Data Analysis:
With the spectrophotometer, we will be measuring the absorbance of light in our sample
and how that relates to protein content. By using standards, or samples with known protein
concentrations, we can plot the absorbance of the standards against their concentration to give us
a standard curve. Then, since we only know the absorbance of the other samples, we can
compare them to the standard curve to determine a very accurate estimate of the protein content
of our samples. We will then compare the average protein contents of the mealworms raised on
polyethylene and the mealworms raised on brown rice to see if there is a statistical difference,
and if so, what does that difference mean.

Chemical Disposal:
Will reference the material data sheet.

Interpretation:
This experiment is comparing mealworms raised on polyethylene and mealworms raised
on brown rice. One will have a higher protein content then the other, but there are a variety of
factors to consider. The timeline of production is one such factor. Brown rice takes 120 days to
reach maturity.14 The mealworm is the larva of the darkling beetle, and their life cycle averages
from three months to a year, with the larval stage being reached after about a month.15 This
means that after about 150 days you will see a return on the brown rice mealworms, as you have
to raise the rice first and then the mealworms, and after about thirty you will see a return on the

14
“Rice plants grow to a height of three to four feet over an average of 120 days after planting.” This was found at
https://www.usarice.com/thinkrice/discover-us-rice/how-rice-grows.
15
“It will take around one to four weeks for an egg to hatch and the larva to emerge.” I found this at https://mealwormcare.org/life-cycle/.
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plastic ones.16 What significance does this have? If the brown rice raised mealworms take five
times longer to grow, it only makes sense that they would need to have five times more protein in
order to make up the difference. This means that our benchmark on the success of polyethylene
raised mealworms is that they need to have at least 20% of the protein of the brown rice ones,
although I believe that they will be fairly close.

Bibliography:
Document Connect.
www.thermofisher.com/document-connect/document-connect.html?url=https://assets.ther
mofisher.com/TFS-Assets/LSG/manuals/MAN0017135_PierceRapidGoldBCAProteinAs
sayKit_UG.pdf.
A Global Food Crisis | World Food Programme. www.wfp.org/global-hunger-crisis.
“How Rice Grows.” Default, www.usarice.com/thinkrice/discover-us-rice/how-rice-grows.
Just a Moment... mealwormcare.org/life-cycle.
Lai, Olivia. “8 Shocking Plastic Pollution Statistics to Know About.” Earth.Org, 20 Nov. 2022,
earth.org/plastic-pollution-statistics.
Stanford University. “Mealworms Provide Plastic Solution.” Stanford News, 10 Jan. 2020,
news.stanford.edu/2019/12/19/mealworms-provide-plastic-solution.

16
I intend for this to be a solution in countries where people are very self-sufficient, as they commonly lack protein in their diets, and they do not
have food security. Additionally, I doubt that this will ever be adopted in first world countries due to the distaste towards the idea.