University of Guyana

Faculty of Natural Sciences

Department of Biology

BIO4112 – Molecular Biology and Biotechnology

Lecturer: Ms. Daniel

Lab Instructor: Ms. Gillette, Ms. Johnson and Ms.
Persaud

Laboratory Exercise # 4
Bread Mold

Anuradha Ramnauth
1037859

Submission: Friday, 13 October 2023

Title: Bread Mold
Aim: To test the effect of temperature on mold growth

Introduction:

Microscopic organisms called molds are crucial to the decomposition of plant and animal
materials (National Institute of Environmental Health Sciences, 2023). Outside, wet, shaded
spots and areas where leaves or other plants are rotting are breeding grounds for mold (National
Institute of Environmental Health Sciences, 2023). As long as there is organic material, moisture,
and oxygen, indoor molds can develop on almost any surface (National Institute of
Environmental Health Sciences, 2023). Spores are microscopic cells that molds release into the
surrounding air when they are disturbed (National Institute of Environmental Health Sciences,
2023). Every day, everywhere-at home, at work, at school, and both inside and outdoors-people
are exposed to mold (National Institute of Environmental Health Sciences, 2023). It is thought
that the main method that humans are exposed to mold is through their breathing (National
Institute of Environmental Health Sciences, 2023). Mold spores and fragments can become
airborne and enter the air we breathe and people may come into contact with mold through food,
such as bread (National Institute of Environmental Health Sciences, 2023).

Bread molds are several species of molds that grow on bread. They develop as a result of mold
spores getting on the bread's surface (Cook, 2018). The fungus consumes the organic ingredients
in bread and other foods. Three frequent bread molds are Penicillium, Cladosporium, and Black
Bread Mold (Cook, 2018). Eating moldy bread is best avoided as certain varieties are safe to
consume while others are not (Cook, 2018). Despite being distinct species, these molds all share
a few characteristics (Cook, 2018). Spores are produced by bread molds, just like by all other
molds (Cook, 2018). Spores are small, often minuscule particles that eventually give rise to fully
grown molds (Cook, 2018). Spores of mold can be found practically anyplace there's organic
matter and moisture (Cook, 2018). They are virtually always found both indoors and outdoors
since they can float through the air or land in food or water (Cook, 2018). Because bread
contains a wealth of organic ingredients, bread molds grow well on it (Cook, 2018). Mold spores
proliferate due to sugar and carbohydrates (Cook, 2018). The kind of spores that are present in
the environment determines the particular species of bread mold (Cook, 2018).

The most prevalent type of bread mold is called Rhizopus stolonifer, or black bread mold (Cook,
2018). When black bread mold is not removed, it typically manifests as hazy green or blue areas
on the bread's surface that eventually turn black and have splotchy centers (Cook, 2018).
Penicillium molds typically show up as pale blue, gray, or fuzzy white patches on bread (Cook,
2018). Unless a person has allergies, unintentionally consuming penicillium is generally not
harmful (Cook, 2018). Penicillium species that create mycotoxins, on the other hand, have been
connected to many malignancies and other diseases (Cook, 2018). The typical appearance of
Cladosporium molds is as dark patches that can be anywhere from deep green to black in hue
(Cook, 2018). The smell of Cladosporium bread molds is strong (Cook, 2018). Mycotoxins can
be produced by Cladosporium molds, hence extended exposure is not recommended (Cook,
2018).

The availability of nutrients, water activity, pH levels, oxygen, temperature, and moisture are
some of the most important elements in creating the circumstances that lead to the growth of
mold on bread (Cook, 2018).

Materials:

9 slices of Bread

Squirt bottles filled with water

Cupboard (dark area)

Rulers

Clear plastic bags

Paper plates

Black Sharpie marker

Apple juice

Lime juice

Dropper

Method:

1. 9 slices of bread were collected

 2. 1 slice of bread was placed in a bag and water and apple juice were squirted onto the
bread, the bag was closed and it was placed in a dark cupboard. 1 slice of bread was
placed in a bag and water and apple juice were squirted onto the bread, the bag was
closed and it was placed in an area with light. 1 slice of bread was placed in a bag and
apple juice was squirted onto the bread, the bag was closed and it was placed in a dark
cupboard. 1 slice of bread was placed in a bag and apple juice was squirted onto the
bread, the bag was closed and it was placed in an area with light. These were the
experiments for the apple juice.
3. 1 slice of bread was placed in a bag and water and lime juice were squirted onto the
bread, the bag was closed and it was placed in a dark cupboard. 1 slice of bread was
placed in a bag and water and lime juice were squirted onto the bread, the bag was closed
and it was placed in an area with light. 1 slice of bread was placed in a bag and lime juice
was squirted onto the bread, the bag was closed and it was placed in a dark cupboard. 1
slice of bread was placed in a bag and lime juice was squirted onto the bread, the bag was
closed and it was placed in an area with light. These were the experiments for the lime
juice.
4. For steps 2 and 3, the experiments with water added are called the wet experiments and
those without water added are called the dry experiments.
5. 1 slice of bread was placed in to a plastic bag and it was placed in a dark cupboard,
another slice of bread was placed in to a plastic bag and it was placed in an area with
light. These two were the control.
6. The experiments were observed after three days, then after six days and then after nine
days.
7. The color and size of the mold that grew were recorded and pictures were also taken.
8. The results obtained were tabulated and the final observation were taken and the results
were compared with the results of another group.

Results:

Days Wet/ Lime Wet/ Lime Dry/ Lime Dry/ Control in Control in
 Juice/ Dark Juice/ Light Juice/ Dark Lime Dark Light
Juice/
Light
Day No molds No molds No molds No molds No molds No molds
1 observed observed observed observed observed observed

Day No molds No molds No molds No molds No molds No molds

3 observed observed observed observed observed observed

Day No molds Blue-grey Very small, No molds Very Very small,

6 observed fungal growth white molds observed small, white molds
around crust white
molds

Day Thick black Thick blue- Small, white Small, Increased Increased
9 powdery black molds with white number of number of
fungal growth powdery thick blue- molds small small, white
with white fungal growth black around white molds with a
hairs on crust around crust powdery the crust molds yellow color
and extending with a yellow fungal on different
to surface of color growth on parts of the
bread emerging part of crust bread
above the
fungal growth
Title: Table Showing the Results of Day One, Three, Six and Nine for the Dry or Wet, and Light
or Dark and Control Experiments with Lime Juice

Days Wet/ Apple Wet/ Apple Dry/ Dry/ Apple Control Control in
Juice/ Dark Juice/ Light Apple
 Juice/ Juice/ Light in Dark Light
Dark
Day No molds No molds No molds No molds No molds No molds
1 observed observed observed observed observed observed
Day No molds 1cm green- No molds 1cm, green No molds No molds
3 observed white fungal observed fungal growth observed observed
growth at two at two spots
places and and small,
small yellow white molds
and white
molds
Day 2 cm, yellow 1cm green- Very 1cm, green Very Very
6 fungal white fungal small, fungal growth small, small,
growth with growth at two white at two spots white white
white spots places, 1 cm molds and small, molds molds
around white fungal white molds
growth and
small white
molds
Day Yellow Small brown Increased 1cm, green- Increased Increased
9 fungal fungal growth number brown fungal number number of
growth with of small growth with of small small,
covered 90% surrounding white surrounding white white
of one side of white fungus molds white fungus, molds molds with
the bread, it at four spots small black a yellow
had small and white fungal growth, color on
brown-green fungal growth 1 cm green different
spots and of varying fungal growth parts of the
white spots sizes at with bread
growing different spots surrounding
several white fungus
 places on the and small
yellow white molds
fungus. The
other side
had small,
white molds
with a yellow
color
Title: Table Showing the Results of Day One, Three, Six and Nine for the Dry or Wet, and Light
or Dark and Control Experiments with Apple Juice

Discussion:

Question 1- The independent variables of this experiment were the apple juice or lime juice
(different pH levels), the light or dark conditions and the dry or wet conditions (addition or no
addition of water to the bread). In an experimental study, an independent variable is one that is
varied or manipulated in order to examine its effects (Bhandari, 2023). The independent variable
is also known as the cause variable (Bhandari, 2023). The apple juice or lime juice, the light or
dark conditions and the dry or wet conditions were the independent variables since these were
manipulated to have an effect on the bread.

Question 2- The dependent variable of this experiment was the slices of bread. A dependent
variable is the variable that changes as a result of manipulating the independent variable
(Bhandari, 2023). The dependent variable is also known as the effect variable and it is contingent
upon the independent variable (Bhandari, 2023). The slices of bread were the dependent variable
since these had an effect due to the different independent variables that were manipulated.

Question 3- Temperature was not a variable that was manipulated or controlled in this
experiment as such nothing can be said on the effect that temperature had on the growth of mold.
However, mold cannot develop into a healthy colony in extremely cold temperatures (Valenti,
2022) Also, while mold may go dormant in extremely cold conditions, they would not
necessarily die (Valenti, 2022). The same principle applies in really hot conditions, since the
majority of molds like and thrive in temperatures between 60- and 80-degrees Fahrenheit
(Valenti, 2022).
Question 4- Based on the results of the experiment, it can be seen that mold grew in both wet and
dry conditions. However, the pace at which bacteria multiply will also increase with rising
moisture levels (Gurleen, 2021). This is due to the fact that more types of mold and bacteria will
be able to develop and become active at higher moisture contents (Gurleen, 2021). Because
various microbes require different amounts of moisture, more bacteria will develop in an
environment with higher moisture levels (Gurleen, 2021). An example of this can be seen in the
Wet/ Apple Juice/ Dark experiment where the mold occupied more approximately 90% of the
bread, this was the largest mold that was observed.

Question 5- pH had a significant effect on the growth of mold on the bread slices in this
experiment. Even though all of the bread slices had growth of mold, the slices of bread with a
lower pH which were the slices of bread that were treated with lemon juice had less mold growth
than the slices of bread with a higher pH which were the slices of bread that were treated with
apple juice. This is due to the fact that while a higher pH can encourage the growth of molds, a
lower pH can assist prevent the formation of mold (Charpentier, 2023).

Question 6- If the amount of water that was added to the bread slices was increased, then this
would increase the moisture of the bread slices Gurleen. As such, this would create a more
favorable environment for the growth of the bread molds and make then active, which would
mean that there would be an increase in the amount of mold growth present on the bread slices
Gurleen. Then, if the amount of water that was added to the bread slices was decreased, then this
would decrease the moisture of the bread slices Gurleen. As such, this would not create a
favorable environment for the growth of the bread molds and make them less active, which
would mean that there would be a decrease in the amount of mold growth present on the bread
slices Gurleen.

Question 7-

There a few similarities and differences when comparing my results and observations to my
classmate’s results and observations.

For lime juice/ wet/ dark, we had the same results, molds were not observed until day 9. We
also had the same result for lime juice/ wet/ light, molds were not observed on day three,
however almost similar molds were observed on day six and nine. For lime juice/ dry/ dark, we
had similar results until day nice, since we both did not observe any molds for day three and
day six. However, for day nine, I observed small, white molds around the crust while they did
not observe any molds. For lime juice/ dry/ light, similarly no molds were observed until day 6.
However, the molds that we observed differed in color, I observed small white molds which
grew into small, white molds with thick blue-black powdery fungal growth on part of crust,
while they observed dark and colored molds.

For apple juice/ wet/dark, we both did not observe any mold on day three, however on day six I
observed yellow fungal growth with white spots around while they did not observe any molds.
Then for day nine, they observed a yellowish mold while the yellow mold that I observed grew
to cover 90% of one side of the bread with small brown-green spots and white spots growing
several places on the yellow fungus and the other side of the bread had small, white molds with a
yellow color. For apple juice/ wet/light they did not observe any mold on day three, while I
observed green-white fungal growth at two places and small yellow and white molds. Then for
day six, I observed green-white fungal growth at two places, white fungal growth and small
white molds while they observed dark colored molds. Then for day nine, I observed small brown
fungal growth with surrounding white fungus and white fungal growth of varying sizes at
different spots while they observed several colored molds. For apple juice/ dry/ dark we both did
not observe molds on day three, and then for days six and nine they observed dark and green
molds while I observed small white molds. For apple juice/ dry/ light they did not observe any
mold on day three while I observed green fungal growth and small white molds. Then for day six
they observed small molds while I observed green fungal growth and small white molds. Then
for day nine they observed several colored molds while I observed green- brown fungal growth
with surrounding white fungus, small black fungal growth, green fungal growth with surrounding
white fungus and small white molds.

For the control/ dark, we both did not observe any molds in day three and then we observed
molds for days six and nine (I observed small white molds). Then for control/ light, they did not
observe any molds while I observed small white molds and then an increased number of small,
white molds with a yellow color on different parts of the bread.
The difference observed between the two results and observations can be to a number of
different reasons and sources of errors. One of such can be the amount of each liquid, lime or
apple juice or water that was added to the bread. The amount of exposure to air that the bread
slices received, this can result if they were not properly closed. The amount of light and darkness
the slices received since they were all placed in different locations. Also, the difference in
temperature of the areas that the different slices were placed.

Conclusion:

While some fungi live freely in soil and water, others collaborate with plants and animals to
develop symbiotic relationships. Different locations support different types of mold growth.
Certain types of organisms thrive on wood, while others eat decaying plant and animal materials.
Additionally, there are those that develop on bread; these are known as bread molds. Bread mold
is a type of fungus that grows on bread and are made up of multiple fungal cells that absorb
nutrients from their environment since they cannot make their own food. Spores of mold can be
found practically anyplace there's organic matter and moisture. They are able to float through the
air and land in food or water. A number of elements influence the formation of molds, and
depending on their particular importance, they can either promote or prevent mold growth. Based
on the results it can be concluded that pH, light and moisture have an effect on the growth of
mold on bread. Bread molds cannot develop in extremely hot nor cold temperatures, they grow
more in wet environments since they favor these environments and are more active, and they
thrive better in higher pH levels since lower pH inhibits their growth.

References:

Bhandari, P. (2023, June 22). Independent vs. Dependent Variables | Definition & Examples.

Scribbr. https://www.scribbr.com/methodology/independent-and-dependent-variables/

Charpentier, R. (2023, April 4). Pro Tip: Understand the 9 factors affecting pH levels in bread

baking. Baking Business. https://www.bakingbusiness.com/articles/58797-pro-tip-

understand-the-9-factors-affecting-ph-levels-in-bread-baking#:~:text=Shelf%20life%3A

%20A%20lower%20pH,fresh%20for%20an%20extended%20period.
Cook, M. (2018, April 11). Different kinds of bread mold. Sciencing.

https://sciencing.com/different-kinds-bread-mold-5956459.html

Gurleen, L. (2021a). The Effect of Moisture on the Growth of Mold on Organic Bread. UBC

Library: Open Access Journal Hosting.

https://ojs.library.ubc.ca/index.php/expedition/article/view/196125/191375

Gurleen, L. (2021b). The Effect of Moisture on the Growth of Mold on Organic Bread. UBC

Library: Open Access Journal Hosting.

https://ojs.library.ubc.ca/index.php/expedition/article/view/196125/191375

National Institute of Environmental Health Sciences. (2023). Mold. National Institute of

Environmental Health Sciences.

https://www.niehs.nih.gov/health/topics/agents/mold/index.cfm

Valenti, R. (2022, January 3). How does temperature affect mold growth. MoldGone.

https://moldgone.com/how-does-temperature-affect-mold-growth/#:~:text=If%20the

%20temperature%20is%20too,between%2060%2D80%20degrees%20F.