Bread_Mold_Experiment

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143

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Biology

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Dec 6, 2023

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Williams 1 Kirsten Williams Dr. Lievens BIOL 144-L 3 April 2020 The Growth of the Fungus Rhizopus stolonifera on Different Bread Types Abstract This experiment was done to show the effect of saprophytic fungi mold on two slices of bread, one with preservatives and the other without. To complete this experiment, two slices of bread were separately placed in a culture bowl with water placed around the edges of the bread. Spores from the fungus Rhizopus stolonifera were added to the center of each piece of bread so that the fungus would spread from the center outward. The culture bowl was covered with plastic wrap and sealed with a rubber band to ensure that the environment was moist enough for the mold to grow, and it was also placed in a cabinet so that there was no light. The bread molds were measured every day for two weeks around the same time. It took a few days for growth to occur in both cultures, but overall the results showed that the bread with no preservatives was completely engulfed in the mycelium, while the bread with preservatives had mycelium growth but not as much. Introduction Rhizopus stolonifera is a fungus known by the name of “black bread mold” because of its ability to contaminate bread and turn it a black or grey color (Barbaneagra, et al. 2011). This fungus needs moisture and darkness in order to grow,

Williams 2 and the darker and moister the environment is, the more it will grow. Because this is a reactive oxygen species (ROS), it will react strongly when no light and little oxygen is present. If Rhizopus stolonifera is placed on a piece of bread containing preservatives and a piece of bread with no preservatives, and both are placed in moist and dark environments, then the bread with no preservatives will appear to have a larger diameter of mycelium than the bread with preservatives. A fungus like this usually takes much longer to appear on any type of packaged bread, and this is through a study about modified atmosphere packaging (MAP). MAP is a way of packaging bread that is modified to have gases that will preserve bread longer (Fernandez 2006). The tight space used in the experiment was to allow the fungus to have optimal growing conditions. Materials and Methods A slice of bread containing preservatives and a culture bowl were obtained and the crust was cut off of the bread to fit in the bowl. A china marker was used to mark the bowl as “preservatives” and drops of water were placed along the edges of the bowl to provide a moist environment. Five drops of Rhizopus stolonifera spores were placed in the center of the bread. The bowl was then covered with plastic wrap and secured with a rubber band. A slice of bread with no preservatives and a culture bowl were obtained and the crust was cut off of the bread to fit in the bowl. A china marker was used to mark the bowl as “no preservatives” and drops of water were placed along the edges of the bowl to provide a moist environment. Five drops of Rhizopus stolonifera spores were placed

Williams 3 in the center of the bread. The bowl was then covered with plastic wrap and secured with a rubber band. Ingredients for both bread samples were provided for further research. The culture bowls were placed in a dark place within the laboratory because temperature was a factor in the growth of the mycelium. Each day around the same time the growth was measured with a ruler by removing the plastic wrap and using a microscope if needed. Observations of growth and color were recorded over the course of the experiment. Results The results of the experiment agreed with the original hypothesis that the bread with no preservatives would have more mycelium than the bread with preservatives. In conclusion, the bread with no preservatives had a mycelium length of 110 mm and the bread with preservatives had a mycelium length of 80 mm. We allowed the mycelium to grow for 14 days, until the mycelium of the bread with no preservatives consumed the petri dish (Figure 1). Although the mycelium on the bread with no preservatives had a greater growth rate, both fungi showed steady growth on the breads after they started growing (Graph 1). Graph 1. Mycelium Growth 0 20 40 60 80 100 120 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Mycelium Length in mm Day Preservatives No Preservatives

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Williams 4 Figure 1. Mycelium Growth on Day 14 Overall, the results of this experiment proved the original hypothesis to be correct because the bread with no preservatives showed more mycelium growth. Discussion The results revealed that the bread with no preservatives built up more mold than the bread with preservatives. The growth started off slow, but then quickly the mycelium started growing on both cultures. Due to the limited amount of space that the culture had to grow, the mycelium on the culture with no preservatives could only grow to the diameter of the container, 110 mm. This experiment showed that preservatives can ultimately prevent the growth of fungi for a little longer than that of bread with no preservatives. It also showed how fungus affects crops and food that is sold and bought in stores. This experiment could also be used on various types of crops to determine how they are affected. The experiment could contain crops sprayed with chemicals and crops naturally grown, with a fungus such as Rhizopus stolonifera implanted in each subject.

Williams 5 Acknowledgments I would like to thank Dr. Lievens for guiding us through this experiment and setting everything up in order for this experiment to be successful. I would like to thank my lab partners, Jaclyn Smolik and Meagan Campbell, for completing this experiment alongside me. References Barbaneagra, et al. “Researches Regarding Catalase and Peroxidase Activity.” University of Agricultural Sciences and Veterinary Medicine , vol. 54, Feb. 2011, pp. 44-49. EBSCOhost . Accessed 20 March 2020. Fernandez, Ursula, et al. “Extended Shelf Life of Soy Bread.” Journal of Food Protection , vol. 69, no. 3, Apr. 2006, pp. 693-698. EBSCOhost . Accessed 20 March 2020. Gustafson, Mark. “Laboratory Manual for Biological Systems II.” Texas Lutheran University Biology Department . 2020.

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