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“Lab 6 Microbial Ecology” “BIO250L”
Student Name: CAROLINE NANSUBUGA
Kit Code (located on the lid of your lab kit): AC-GRJPW01
“Experiment 1- Mold Growth on Bread and Fruit- What are Optimal Conditions for Fungal Growth?”
“Table 1: Growth Results”
“Condition”
“Extent of Growth”
“Day 2”
“Day 4”
“Day 6”
“Day 8”
“Day 10”
“Day 12”
“Day 14”
“Bright- Control”
0
0
0
2
3
1
1
“Bright- Water”
0
1
1
1
2
3
3
“Bright- Sugar”
0
0
0
0
0
2
3
“Bright- Lemon”
0
0
1
1
2
2
2
“Bright- Vinegar”
0
0
0
0
0
2
2
“Dark- Control”
0
1
3
3
4
4
4
“Dark- Water”
0
0
1
1
2
2
3
“Dark- Sugar”
0
0
1
1
1
2
3
“Dark- Lemon”
0
0
0
0
0
0
1
“Dark- Vinegar”
0
0
0
0
0
1
2
“Table Key:” “0 = no growth”
“1 = little/some growth (1-25%)”
“2 = moderate growth (26-50%)”
“3 = heavy growth (51-75%)”
“4 = extensive growth (76-100%)”
“Post-Lab Questions”
“1. Which condition produced the most mold growth? The fastest? The least? The slowest?”
Dark control produced the most and fastest mold growth. Dark lemon produced the least. Dark vinegar produced the slowest.
“2. Based on your results, what conditions are most favorable for mold growth on bread and apples?”
The conditions most favorable for mold growth on bread and apples are, temperature, pH, nutrients, water activity, and aeration.
Temperature; since most mold are mesophillic, meaning, they can grow at a temperature range of 10-35 degrees celsius. Therefore, optimum temoerature for mold growth ranges between 15-35degrees celsius.
pH; This varies from mold to mold. But most mold growth are favorable with a pH range of 3-7. For example, niger, aspergillu. Penicillium is the only one capable of growing at a pH of 2 and below.
Nutrients; For growth, different molds will require different nutrients. Some will grow well with simple sugars, others with complex sugars, and the rest will prefer nutrients with high sugar or salt.
Water/moisture; This one is a necessity for all molds to grow. They all need moisture though the amount required varies from mold to mold. For example, xerophile can grow under low moisture, whereas, hydrophillic need high moisture to grow.
Aeration; Air is a necessity for all molds to grow.
“3. Would these conditions apply to all fungal growth? Why or why not?”
Yes, they would apply to all fungal growth, although, the amount of each condition needed varies from fungal to fungal as I explained in the previous question.
“4. How would changing the type of bread (fresh from a bakery, no preservatives vs. prepackaged with preservatives) affect the results? Describe an experiment that would test your hypothesis.”
Since some preservatives are anti-fungal in nature, changing the type of bread may affect growth rate. Even though Fungi grows best in the moist-dark habitants, they can also grow anywhere else with organic material. Most fungi are saprophytes, which means that they are securing their nutrients from dead organic material. Fungi, like any other bacteria releases hydrolytic exoenzymes which digest external substrates. In the process, absorbing soluble product. As a source of carbon,energy, and electrons, they use orgaic compounds . Rhizopus stolonifer, a bread mold, falls under this division. It grows on the surface of moist, carbohydrate rich foods like, vegetables, fruits, and bread. For example, on bread, Rhizopu's hyphae quickly covers the entire surface, forcing Rhizoid (special hyphae) to extend into the bread in order to absorb nutrients.
“5. How would changing the incubation temperature of the bags affect the results? Describe an experiment that would test your hypothesis.”
Mold would grow slower in cold temperatures than in warm temperatures. Get two pieces of bread, any type of bread, consider the date the bread was baked. Place each piece in the zip lock bag and place one bag in a warm place like on top of the refrigerator, and the other piece in the cold place like a closet. Check on the samples every other day for 2 weeks and record the amount of mold growth on both samples.
“6. Look up the pH of lemon juice and vinegar. Based on your results and your knowledge of favorable environmental conditions for fungal growth, what can you conclude about the effect of pH on growth? How would making the pH more basic affect growth?”
Moderately acidic pH promotes growth, whereas extreme low pH like vinegar hampers growth. Making the pH more basic may increase mold growth.
“7. What is the source of the mold that grew on the samples?”
The source of mold growth is fungal pores present in air.
“Insert photo of your cultures after incubation with your name clearly visible in the background:”
“Experiment 2- Fluid Thioglycollate Medium to Assess the Effect of Oxygen on Bacterial Growth”
“Table 2: Experiment 2 Results”
“Sample Location”
“Growth Location in FTM”
“Oxygen Category”
“Control”
Medium
Absence of oxygen
“Skin”
Deep
Presence of ixygen at the top
“Nose”
Numerous growth
No oxygen
“Throat”
Numerous growth everywhere
No oxygen
“Shoe”
Deep
Presence of Oxygen at the top
“Post-Lab Questions”
“1. What does the pink band in the FTM indicate?”
Presence of oxygen.
“2. What would be the effect of using FTM tubes with broad pink zones?”
FTM medium contains a reducing agent, sodium thioglycollate that reduces oxygen to water if it is left go boil for 5 minutes. Making sure yhe cap is tight enough, let it cool at room temperature in order for oxygen to diffuse back into the tube. Resulting into a small pink layer at the top of the broth.
“3. Provide an explanation as to why bacteria from each location displayed its pattern of growth?”
Thioglycolate broth being a multi-purpose and differential medium, it primarily determines the amount of oxygen required for microorganisms. In the process, sodium thioglycolate in the medium consumes oxygen and permits the growth of obligate anaerobes, making bacteria from each location to display its unique pattern of growth.
“4. Why is it important to pull the inoculating needle out of the FTM via the same path you inserted it?”
To avoid inlet and outlet of more oxygen that might change the results.
“5. How would you explain the presence of bacterial growth in the control FTM tube?”
It can be explained in terms of contamination of either the agar or the inserting needle.
“Insert photo of your cultures after incubation with your name clearly visible in the background:”
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