How does moisture level affect mold growth in a warm, dark environment? Abby Olson Calee TeGrotenhuis Meaggan Horton Rachel Hamblin

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1 How does moisture level affect mold growth in a warm, dark environment? Abby Olson Calee TeGrotenhuis Meaggan Horton Rachel Hamblin

2 Prediction We predict that higher moisture levels will be conducive to higher mold growth in a warm, dark environment. We predict that the most mold will grow on the bread sample with the most water added to it in our experiment. We predict this because in using past experiences, we have found that mold levels increase as the moisture levels increase, so the more moisture there is in the environment, the more likely that mold would grow on the subject in that environment. In our prediction, more moisture would reside in the sample with the most water added to it; therefore, that sample with the most water will grow the most mold.

3 Procedure Materials: 5 slices of 35% whole grain white bread 5 petri dishes 5 ziploc bags 1 paper bag square grids Water 1 graduated cylinder 1 pipette Tape 1 Scissors 1 black Sharpie marker 1 incubator

4 Procedure Variables Constants Condition Temperature Incubator Light Inside a paper bag Type of bread 35% whole grain white Surface swipe n/a Independent Variable Condition Moisture Level 0 ml, 4 ml, 8 ml, 12 ml, 16 ml Dependent Variable Condition Mold growth?

5 Procedure 1. Collect all materials at lab stations. Take out five Petri dishes, a graduated cylinder, a pipette, water and tape. Take apart each Petri dish so there is a top lid and a bottom of Petri dish container. Take the Petri lids and place them next to their bottom part. Take one plastic grid and tape it onto the lid of one Petri dish. Repeat this procedure with the remaining four Petri dish lids and plastic grids. Set these lids aside. 2. Designate only one group member to handle the bread in this experiment. Take one plastic square grid containing 144 total small squares and place it on top of a slice of Sarah Lee 35% whole grain white bread. Use scissors to cut the bread around the plastic grid, creating a square piece of bread. Immediately place the bread square into the Petri dish to assure it does not touch the table. Repeat this process with the remaining pieces of bread.

6 Procedure (continued) 3. Take out a graduated cylinder, a pipette, and water. Take out one Petri dish bottom with its bread square. Using the graduated cylinder, measure 4 milliliters (ml) of water, you may need to use the pipette to add droplets of water to ensure accuracy. Pour the water over the bread in the Petri dish. Cover the dish with a lid and close the container. Place the closed container in a plastic Ziploc bag and seal it. Use a black Sharpie marker to write 4 ml on the plastic bag. Place the plastic bag and Petri dish into the brown paper bag and label the bag with your group s initials. Take out another Petri dish bottom and its bread square. Using the graduated cylinder, measure 8 ml of water. Pour the water over the bread in the Petri dish. Cover the dish with the lid and close the container. Place the closed container in a plastic Ziploc bag and seal it. Use a black Sharpie marker to write 8 ml on the plastic bag. Place the plastic bag and Petri dish into the brown paper bag. Take out another Petri dish bottom and its bread square. Using the graduated cylinder, measure 12 ml of water. Pour the water over the bread in the Petri dish. Cover the dish with the lid and close the container. Place the closed container in a plastic Ziploc bag and seal it. Use a black Sharpie marker to write 12 ml on the plastic bag. Place the plastic bag and Petri dish into the brown paper bag. Take out another Petri dish bottom and its bread square. Using the graduated cylinder, measure 16 ml of water. Pour the water over the bread in the Petri dish. Cover the dish with the lid and close the container. Place the closed container in a plastic Ziploc bag and seal it. Use a black Sharpie marker to write 16 ml on the plastic bag. Place the plastic bag and Petri dish into the brown paper bag. Take the remaining Petri dish bottom and its bread square and cover with the remaining lid so the container is closed. Place the closed container in a plastic Ziploc bag and seal it. Use a black Sharpie marker to write 0 ml on the plastic bag. Place the plastic bag and Petri dish into the brown paper bag. Close brown paper bag.. *This will be the experiment s control since there is no water on the bread.

7 Procedure (continued) 4. Set the brown paper bag with the Petri dishes next to each other in the incubator and leave them there until it s time to check them each week. Check on them at the same time every Tuesday and Thursday for four weeks. 5. Clean up the work area. Put all cutting utensils, tape, markers, and extra plastic grids away. Clean up any water spills. Wash and dry graduated cylinder. Throw away any extra bread. 6. When checking on the Petri dishes, gently take the plastic bags out of the paper bag. Move the Petri dishes in the plastic bags minimally, and do not shake the Petri dishes. Observe the mold on the square piece of bread. In this experiment, the amount of mold is measured in squares covered. Record how many squares of mold, if any, has grown on each piece of bread. If you spot mold in any square, count that square. Record the amount of mold in a data table like the example shown below. Date Amount of water measured in milliliters (ml) 0 ml 4 ml 8mL 12mL 16 ml 2/15/11 0 sq. 0 sq. 0 sq. 0 sq. 0 sq. 2/17/11 2/22/11 2/24/11

8 Procedure (continued) 7. Also record the qualities of the mold each check in day. Include mold color, any shapes the mold may be taking as it spreads over the bread, and any other qualities that may be noteworthy. Record this data in a data table like the example shown below. Date 0mL 4 ml 8 ml 12 ml 16 ml 3/15/11 No change No change No change Light green patches Light green patches 3/17/11 No change No change Blue and gr. areas Light green with Black sink hole, white fibers yellow circles 8. Once the amount of mold is measured each day, carefully place the plastic bags into the brown paper bag each time and close the paper bag. Place the brown paper bag back into the incubator after every observation. 9. At the end of four weeks, after recordings are finished, clean up the experiment components. Clean out the Petri dishes that do not have much mold on them and throw away the Petri dishes that are 60% or more covered in mold. Throw away the plastic bags and paper bags used in this experiment.

9 Mold Growth Number of Squares with Mold ml 4ml 8ml 12 ml 16 ml Date

10 Date 0 ml 4 ml 8 ml 12 ml 16 ml 2/15/11 No change No change No change Light green patches 2/17/11 No change No change Green and blue areas 2/22/11 No change Small green and blue areas 2/24/11 No change Green spot with white hair fibers surrounding 3/1/11 No change Dark green spot in the middle 3/3/11 No change Dark green and orange spot with white hair fibers 3/8/11 No change Green center, brown corner, white hair fibers Dark blue spot in the middle Dark green sink hole Dark green sink hole Giant dark green sink hole Giant dark green sink hole, yellow corner (looks mossy) Light green with white fibers Light green with white fibers Mint green with white fibers Light green, brown spot at top Brown spot now orange with a lot of white fibers A lot of white fibers, yellow and orange sides, sink hole under the fibers Light green patches Light green and yellow circles Black sink hole, yellow circles surrounded by red and purple, white fibers Dark black sink hole Dark black sink hole, yellowing of corners Bread has turned to mush Bread is green, yellow, orange, black and soupy

11

12

13 Claim At the conclusion of our experiment we can conclude that moisture facilitates the growth of mold. This is evidenced by our data that showed that the more moisture in the petri dish the faster the mold appeared and grew.

14 Reflections During our experiment we made a few errors: Counting both sides of the bread Holding up the bread/moving it around (See picture) Grid squares not lined up with bread Perhaps our variable of moisture had an interval that was too small.

15

16 References to help support our claim include: Mold requires moisture in order to grow. That is why cereal doesn't grow mold but bread in a bag does. Bread has enough moisture to allow the mold to grow. If you make cereal soggy it will grow mold. David Beck With adequate moisture and nutrients from the bread, this spore sprouts and grows hair like structures on the bread surface. Once the mold attains a particular growth with paint brush like structures, it starts producing fruiting structures. Josh Riverside The optimal growth conditions for fungi are warmth and moisture. Once spores infest bread, mold will grow unless it loses all the moisture in its surroundings. Peter De Conceicao

17 References Beck, D. (1999, May 27). Bread mold project with rhizopus spores. Retrieved March 20, 2011, from MadSci Network: 06/ Mi.r.html Conceicao, P. D. (2010, June 27). How Does Light Affect the Production of Mold? Retrieved March 20, 2011, from ehow: reproduction mold bread_.html Riverside, J. (n.d.). Bread Mold. Retrieved March 20, 2011, from Ezine Articles: Mold&id=405845

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