Douglas_Hannah_Biofilms

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Hannah Douglas Biology 159 M. Bunch 10/06/2022 The pH levels in Galactose and Ethanol biofilms Introduction Microbial biofilms are populations of microorganisms and survive in varying environments. In a biofilm the cells adhere to each other, they also surface into an environmental signal. These can be single or multi species by grouping of cells, they are covered in a slime like layer. This slime layer provides an environment for many cells to group together to create a biofilm with multi species. Biofilms can be found on almost all non- shedding solid moist surface such as pipes, teeth, catheters, and many more (Bacterial Biofilms 2004). There are two different pathways for the carbon metabolism within the biofilm. The first is with oxygen present and the second is without oxygen present. Biofilms are most often referred to as microorganisms or bacteria (Discovering Biofilms 2012). These bacteria adhere to a solid surface and adapt to environment changes of the surface. When biofilms are without oxygen, they are called fermentation. The biofilm produces less energy per molecule of carbon. This still maintains survival for the biofilm in anaerobic environments. Only smaller organisms can survive on fermentation alone. After we reviewed this information on the fermentation of biofilms, we began to question the levels and morphology of the biofilms in different stages of fermentation or oxidation. After we asked these questions, we began to think about the effect that fermentation has on pH levels. Which led us to our hypothesis of, the presence of a fermentable carbon source effects the pH level of a yeast strand because an acid is produced during fermentation. Then we predicted that Galactose causes a higher increase in the pH levels of yeast than Ethanol. We considered that Galactose is a sugar with a low amount of oxygen which led us to believe that the pH levels would be higher with the presence of the yeast. Therefore, the null hypothesis was that there will be no difference in the pH levels of Galactose and Ethanol when fermented in the yeast. Methods Experimental Design In this experiment, the independent variable was the Carbon source (ethanol and galactose) we used both a fermentable and non-fermentable carbon source. The second independent variable was the yeast strain (TRY 138 WT, TRY 140 FLO DOWN, TRY 143 FLO UP). The dependent variable was the pH levels. We gathered a total of 7 plates for this experiment, we used 3 Galactose plates, 3 Ethanol plates, and 1 control plate with glucose. We placed each yeast strain in each of the dishes. We had the dishes marked off for each strain labeled by the name of each strain. Each dish was touched with a toothpick with each yeast strain. The dishes were labeled with a group members name as well as the type of plate it was. These dishes were left for a week and then we tested the pH levels of the 3 yeast strains of each plate. Data Collection After the week period, we measured the pH levels of each of the yeast strains. There was minimal difference between the pH levels of the Galactose and the Ethanol. However, overall, the Ethanol had higher pH than the Galactose. After collecting this information, we measured that the data did not support our hypothesis and the pH was higher in the Ethanol dishes than in the Galactose dishes. We also compared the pH levels to determine a p value of .05. Results Question: How does fermentation effect the pH levels of yeast?

Hannah Douglas Biology 159 M. Bunch 10/06/2022 Figure 1 Strain 1- 138 Sugar A- Galactose Strain 2- 140 Sugar B- Ethanol Strain 3- 143 Comparison of the pH levels in both of the sugars. In this graph it shows the comparison of the levels of pH in all the yeast strains as well as the different sugars. The pH levels are overall higher for sugar B (Ethanol). The Ethanol created an environment for increase pH in all the strains of yeast. This shows that Galactose limited the environment for pH resulting in an overall lesser amount of pH in all of the Yeast strains. Discussion Hypothesis Considering the results of our experiment we were able to assess that our hypothesis was incorrect. Our hypothesis was that the pH levels of the yeast would be higher in the Galactose than in the Ethanol. Our results show that the pH levels of the yeast were higher in the Ethanol than in the Galactose. Consistently the pH in the control remained the same and did not increase. Both of the sugars increased the pH; however, the fermentable sugar, Galactose, created less of an increase than the non-fermentable sugar, Ethanol. Analysis of Data Although the data we collected was accurate, there is always room for error. There is always a place for human error when measuring or inoculating. The contamination of fruit flies for this experiment could have affected the data that we collected. There was many different shapes and often other growths on our dishes, this making us believe there was contamination within our experiment. Implications After we researched more, we learned that during fermentation the cells take in other ions therefore, the pH does not increase as much. The fermentation is creating a space for the levels not to increase and the sugar that is non-fermentable cannot do that. Further Investigation In the further, I would like to research how fermentation and non-fermentation effect the growth on the yeast strains. Is it consistent with the increase of pH levels? References cited Bacterial Biofilms: From the Natural Environment to Infectious Diseases, 2004 Discovering Biofilms: Inquiry-Based Activities for the Classroom, 2012

Hannah Douglas Biology 159 M. Bunch 10/06/2022 Skills of Biological Investigation Biology 159 Laboratory Manual 3E, 2022

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