Lab Report Wk 5

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Kiriri Women’s University of Science and Technology *

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Nov 24, 2024

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1 Oil Spill Bioremediation Laboratory Naamiya Warner SCI 207: Our Dependence Upon the Environment Instructor’s Name: Dr. Kuchanur Date: September 25th 2023

2 Oil Spill Bioremediation Introduction "Bioremediation" employs green plants' enzymes or decomposers to clean up polluted settings. Bacteria may be utilized to clean up oil spills in the water through bioremediation. Hydrocarbons, found in oil and gasoline, are one type of specialized contamination that may be bioremediated using certain bacteria (Luo et al., 2021). Seabirds and other filter feeders are particularly negatively impacted by oil leaks in the water. Like ducks and gulls, seabirds devote most of their lives in the water only to come ashore to lay their eggs (Mishra et al., 2020). Many seabirds have wettable plumage that needs to be thoroughly preened (or dried) before flying. When oil comes into touch with feathers, the seabird attempts to preen while ingesting the oil. Clams and oysters are examples of filter-feeders that filter surface water through their gills to remove any tiny food. If there is oil in the sea, it concentrates inside these shellfish and subsequently builds up in greater amounts in their predators. It is crucial that biologists be aware of oil leaks and that they get fixed up quickly to reduce any harm to people and the ecosystem. The bioremediation technique, which involves bringing microbes or other living things to a damaged region either naturally or deliberately to consume and degrade ecological toxins, will be employed in this lab. The idea is to mimic the bioremediation of an oil leak in the ocean. Adding a chemical substance and an oil-degrading bacteria mixture to small amounts of oil in test tubes aims to examine the utilization of organisms to assist in cleaning up environmental contamination. The occurrence of a hue shift indicates an imbalance in the chemical composition of the oil.

3 Hypotheses Oil Spill Bioremediation activity hypothesized that Tube 1's color will remain the same because of its large amount of water if it contains 1 mL of tetrazolium, 4 mL of filtered water, and ten drops of frying oil. In contrast, the color will alter if tube 2 contains 1 mL of tetrazolium, 2 mL of filtered water, ten drops of frying oil, and 2 mL of the microbial mixture because a nonclear component has been introduced. In Tubes 3 and 4:cooking oil and water are incompatible; therefore, I hypothesized that if tube 3 contains 5 mL of water and ten droplets of cooking oil, the oil detaches from the water, and the bulk of the liquid will remain clear. In contrast, the oil is broken down if tube 4 contains water, oil, and microbial mixture since the microbial mixture will break down the oil, and the liquid will alter coloration. Finally, if tube 5 contains water, oil, and a microbial mixture and is kept at an elevated temperature, the decomposition of the bacteria will be more pronounced since the heat will speed up the decomposition. On the other hand, bacteria will gradually break down in tube six if it contains water, oil, and a microbial solution and is kept at a lower temperature since the cooler temperature will delay the decomposition process.

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4 Results Data Tables and Observations Data Table 1. Tubes 1 and 2 Observations Day Tube 1 Tube 2 0 (Initial setup) The transparent liquid is developing an oil band at its highest point and has some bubbles. The liquid is yellow-tinted and murky, and an oil ring develops on the surface. 1 The oil band at the apex and a few bubbles are still visible in the liquid, which is still transparent. A small amount of the microbial mixture has settled at the tube's bottom, and the liquid is less hazy but retains a slight yellow hue. An oil band has also developed on the highest point of the liquid. 2 The liquid remains transparent even with the oil band on the highest point of the liquid and five different-sized bubbles in the tube walls. Even while the liquid is less hazy than it was on Day 1, it has an oil band on top, a yellow tinge, and a deposited microbiological mixture at the bottom. 3 The liquid remains transparent even with the oil ring developed at the highest point. Although the liquid is transparent, it remains foggy and has a yellow color. Additionally, the accumulated microbial mixture at the bottom and the oil band at the top remain discernible. Data Table 2. Tubes 3 and 4 Observations

5 Day Tube 3 Tube 4 0 (Initial setup) The liquid is transparent and has an oil band growing on the surface. An oil band develops at the top of the hazy, yellowish liquid. 1 The liquid remains transparent and has a top- formed oil band. A small amount of the microbial mixture has deposited at the tube's bottom, and the liquid appears less hazy but has a slight yellow hue. An oil band has also developed at the top. 2 The oil band has formed at the top of the liquid, which remains transparent. Even while the liquid seems less hazy than it was on Day 1, it retains a yellow hue, an oil band on the top, and a microbiological mixture that has dropped to the bottom. 3 The liquid remains transparent and has a top- formed oil band. Even though the liquid retains a yellow hue and is hazy, it is transparent. Additionally, the accumulated microbial mixture at the bottom and the oil band at the surface remain discernible. Data Table 3. Tubes 5 and 6 Observations Chosen environmental factor to change: Change in temperature (hot outdoors and chilly inside a refrigerator)

6 Day Tube 5 Tube 6 0 (Initial setup) An oil band is developing at the highest point of the liquid, which is quite hazy and has a yellow tinge. An oil band is developing at the surface of the liquid, which is quite hazy and has a yellow tinge. 1 Less foggy and colored with a yellow tinge, the liquid has an oil band at the surface, and part of the microbiological mixture is embedded at the tube's bottom. A small amount of the microbial mixture is now settled at the tube's bottom, and the liquid remains quite hazy and has a yellow hue. An oil band has also developed at the very surface. 2 In addition to an oil band at the surface and part of the microbial mixture settling at the tube's bottom, the liquid remains hazy and has a little yellow color. Although very hazy and tinted yellow, the liquid has an oil band at its surface, and part of the microbiological mixture is embedded at the tube's bottom. 3 Although the liquid has a faint pink and yellow hue, it remains highly hazy due to the oil band at its surface and the microbial mixture that has accumulated at the tube's bottom. Even though the oil band at the surface and the microbiological mixture at the bottom of the tube have sunk, the liquid remains a faint pink color and is highly hazy. The darkest liquid is in Tube 6. Graphs and Photographs Figure 1: Day 1

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7 Figure 2: Day 2 Figure 3: Day 3

8 Analysis For tube 1, I hypothesized that adding 1 mL of tetrazolium, 4 mL of purified water, and 10 drops of oil would result in an unchanged color due to the high concentration of water present. Because the liquid remained clear, the data supported my idea. For tube 2, I hypothesized that adding 1 mL of tetrazolium, 2 mL of purified water, 10 drops of oil, and 2 mL of microbial mixture would result in the addition of a nonclear substance, causing the coloring to shift. I accepted my idea since the liquid had altered color, and the findings revealed that it got hazy with a yellow tinge, and part of the microbial mixture sank at the bottom. For tube 3, I hypothesized that since water and oil are incompatible, the mixture would split into the two components, with the water making up the vast majority of the liquid and remaining clear. I confirmed my theory for tube three since the findings showed that the oil did not combine with the water, and the liquid remained pure. In tube 4, I proposed that when water, oil, and microbial mixture are combined, the oil will degrade, and the liquid will take on a different color due to the microbial mixture's disintegration of the oil's characteristics. Since the data demonstrated that the liquid did acquire a yellowish, murky hue, and part of the microbiological mixture eventually settled to the bottom, I

9 accepted my premise. For tube 5, I predicted that if water, oil, and the microbial mixture were combined and kept at an increased temperature, there would be more microbial decomposition since the heat would hasten the chemical reaction. The experiment revealed that the liquid became extremely turbid and tinted yellow. An oil band also developed at the surface, and part of the microbial mixture sank at the bottom of tube 5 more quickly than tube 6 did. I thus embraced my theory. Lastly, in tube 6, I hypothesized that combining water, oil, and a microbial solution in a cooler environment would delay the bacteria's decomposition since the colder environment would slow the procedure's progress. The outcomes demonstrated that it broke down more slowly than tube 5 and maintained the deepest hue in liquid form. Therefore, my theory is correct. Discussion While working on this lab, I learned a lot about oil spills, including the fact that they may happen accidentally or deliberately. Marine oil leakages are extremely tragic occurrences that significantly threaten the ecosystem (Luo et al., 2021). Despite massive vessel spills like the Exxon Valdez occurrence being frequently blamed for oil leaks that impact marine ecosystems, most oil pollution results from surface runoff, oil transit, port operation, and illegal wastewater emissions (Mishra et al., 2020). Although the natural ecosystem affected by the leak is endangered by the numerous hazardous chemicals in the oil, most native, organic microorganisms can and even survive the breakdown of these harmful substances. Bioremediation is the practice of employing microbes for such restoration operations, and it is an effective way of cleaning marine regions harmed by oil spills. I encountered no difficulties doing this lab. With just one activity, it was incredibly easy. The several observation days were challenging because the beverages appeared almost identical.

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10 Because of this, I found it difficult to convey the distinctions, if any. Are there any poisons in the cleaning that can adversely affect the surroundings and living things? In light of what I learned from this testing, there is a query regarding the subject. I would carry out the experiments outlined in this lab's treatment recommendations and then, if possible, examine the results using an organic mixture or another substance devoid of potentially dangerous components. References Luo, Q., Hou, D., Jiang, D., & Chen, W. (2021). Bioremediation of marine oil spills by immobilized oil-degrading bacteria and nutrition emulsion. Biodegradation , 32 (2), 165– 177. https://doi.org/10.1007/s10532-021-09930-5 Mishra, B., Varjani, S., Kumar, G., Awasthi, M. K., Awasthi, S. K., Sindhu, R., Binod, P., Rene, E. R., & Zhang, Z. (2020). Microbial approaches for remediation of pollutants: Innovations, future outlook, and challenges. Energy & Environment , 32 (6), 1029–1058. https://doi.org/10.1177/0958305x19896781

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