SCI207 Week 5 Lab Report - Kimberly Selby

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University Of Arizona *

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207

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Biology

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Jan 9, 2024

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1 Week 5 Lab: Oil Bioremediation Kimberly Selby SCI 207: Our Dependence Upon the Environment Marc Hytnka 11/04/2023

2 Oil Bioremediation Introduction Marine ecosystems and oceans must be protected from oil spills. Wildlife habitats are affected, fish and bird populations are damaged, and the local population is negatively impacted by oil pollution. Among the economic impacts are long-term effects on fish populations and the cost of long-term cleanup. Researchers are exploring bioremediation as a potential remedy for oil spills. “Bioremediation is one technique that may be useful to remove spilled oil under certain geographic and climatic conditions…is defined to include the use of nutrients to enhance the activity of indigenous organisms and/or the addition of naturally occurring non-indigenous microorganisms” (Venosa, n.d.). The objective of this oil spill bioremediation lab is to provide a better understanding of the challenges involved with oil spill cleanups, and how bioremediation can help. Additionally, this lab will test microbes in a variety of environmental conditions. For tubes one and two, there will not be a color change in tube one being that it is only water mixing with the tetrazolium chloride. However, there will likely be a color change in tube two due to the tetrazolium chloride mixing with the microbial solution. My reasoning is that the chemicals in both mixtures will end up slightly combining before all of the elements settle in each tube. For tubes three and four, there will not be a change in the appearance and physical properties of the oil in tube three because water and oil always separate. So, the oil will sit on top of the 5 mls of water. However, I think there will be a change in the appearance and physical properties of the oil in tube three because it will be mixed with whatever the microbial solution is composed of.

3 For tubes five and six, the environmental factor I chose to alter is the temperature. Instead of using 2 mls of room temperature water like the other tubes, I will use 2 mls of ice cold water in tube five and 2 mls of hot water in tube six. I believe that tube six will experience a greater breakdown of oil because of how the microbes react to warmer temperatures. Results Data Tables and Observations Graphs and Photographs

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5 Analysis For tubes one and two, I do not think there will be a color change in tube one because it is only water mixing with the tetrazolium chloride. However, I think there will be a color change in tube two due to the tetrazolium chloride mixing with whatever the microbial solution is composed of. When I first set up the experiment and let the tube components settle, tube one’s liquid layer was clear, and there was a thick, flat layer of oil on top that had no bubbles. When I tipped tube one horizontally, the oil stayed together as a thick layer over the liquid layer and moved slowly from side to side. When I set the tube back upright in the rack, the oil seemed to coagulate to one side of the tube and turned cloudy in color. The top of the oil layer now looked concave, and the liquid layer remained clear. Initially, with tube two, the liquid layer was cloudy and yellowish in color, with a thick, bubbly oil layer on top. When I tipped tube two horizontally, the oil moved over the liquid in what looked like individual oil bubbles, that did not conjoin when they touched each other. The oil layer moved freely and fast from side to side, looking bouncy. When I set the tube back upright in the rack, the oil layer went back up to the top of the liquid layer and remained a thick, bubbly layer, consisting of tiny individual bubbles. The liquid layer also remained cloudy and yellowish in color. For tube one, on day one of observation, the liquid layer was still clear, and the oil layer was still coagulated to one side of the tube and was cloudy and yellowish in color. The top of the oil layer still looked concave. When tipped horizontally, the oil layer stayed together as a thick layer over the liquid layer, with some individual oil bubbles present within the thick layer. Part of the oil layer also looked like it was clotted together over the liquid layer, where I observed that it looked like it was causing a dent in the liquid layer. The oil still moved slowly

6 from side to side and looked bouncy. When I placed it back upright in the rack, the oil layer coagulated back to the side of the tube. However, instead of being cloudy this time, it is mostly clear. The top of the oil layer went back to looking concave. The liquid layer remained clear. For tube two, at the bottom of the tube, there was a pink substance that sunk to the bottom. The liquid layer above it was cloudy and more yellow than the day before. The oil layer is still bubbly; however, today, it looked like it was two different colors; the original color of the oil and a hue of the pinkish substance found at the bottom of the tube. When tipped horizontally, the oil still presented itself as individual bubbles; however, when I held it up to the light, I could see that when the individual oil bubbles touched each other, they would conjoin. They did not do this the day before. The pink substance that was at the bottom of the tube ended up mixing with the liquid layer. When I put the tube back upright in the rack, the liquid layer was more cloudy and pinkish in color. The oil went back up to the top as a bubbly layer, and it still looked like it was two different colors, however; this time, the colors are the original color of the oil and a brownish hue. The pink substance at the bottom of the tube was no longer present as it mixed with the liquid layer. For tube one on day two of observations, the liquid layer was still clear, and the oil was still coagulated on the side of the tube and still looked concave. When tipped horizontally, the oil layer remained thick, with some individual oil bubbles present in the thick layer. Part of the oil layer looked clotted together, which looked like it was causing the liquid layer to dent. The oil moved slowly from side to side, and it looked bouncy. When the individual oil bubbles touched each other, some would conjoin. When I put the tube back upright in the rack, the oil coagulated back to the side of the tube and went back to being cloudy and yellowish in color. The top remained concave, and the liquid layer also remained

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7 clear. For tube two, at the bottom of the tube, the pink substance settled again. The liquid layer above it was pinkish-yellow in color. The oil layer was still bubbly, with medium and small-sized bubbles, and still looked two different colors; the original color of the oil and a hue of the pinkish substance found at the bottom of the tube. When tipped horizontally, the oil still presented itself as individual bubbles, and when held up to the light, I observed that the individual bubbles would conjoin when they touched. The pink substance at the bottom of the tube mixed with the liquid layer, making it cloudy again and more pink in color. The liquid layer was more cloudy and pinkish when I put it back upright in the rack. The oil layer returned to the top and still looked two different colors; the original color of the oil and a pinkish hue. The pink substance at the bottom of the tube was no longer present because it mixed with the liquid layer. For tube one on day three of observations, the oil layer was still coagulated on the side of the tube, and it was yellowish in color. There were also some individual oil bubbles all along the sides of the tube. The top still looked concave, and the liquid layer remained clear. When tipped horizontally, the oil layer still looked like it was staying together, with some individual oil bubbles present within the thick layer. Part of the oil layer also looked clotted together, which looked like it was causing a dent in the liquid layer until I moved it from side to side, breaking apart. The oil moved slowly from side to side and was bouncy; I noticed that some of the individual oil bubbles conjoined when they touched each other. When I put it back upright on the rack, the oil layer coagulated back to the side of the tube and was still yellowish. Individual oil bubbles remained on the sides of the tubes, and the liquid layer remained clear. For tube two, the pink substance settled at the bottom of the tube again. The oil layer was bubbly, with the two sizes of the bubbles being small and large. The color of the oil layer was yellowish, and it also

8 had some pink in it from the substance at the bottom of the tube. The liquid layer was yellowish-pink in color, and it was cloudy. When tipped horizontally, the pink substance mixed with the liquid layer, making it more cloudy and pink. There seemed to be very little oil, as there were only a few individual oil bubbles compared to previous days. There was also some slumps of the pink substance still present that did not mix with the liquid layer. When I put it back upright on the rack, the oil layer returned to being bubbly. However, the size of the oil bubbles was only small. It did not look like there was any pink in the oil layer anymore. The liquid layer was more cloudy and more pink than yellow in color. A few tiny clumps of the pink substance were at the bottom of the tube. Given the results of this experiment and the observations I made along the way, I accept my hypothesis of there being no color change in tube one because tube one only had water mixing with the tetrazolium chloride, and that there would be a color change in tube two because the tetrazolium chloride would mix with whatever the microbial solution is comprised of as being correct. For tubes three and four, I do not think there will be a change in the appearance and physical properties of the oil in tube three because water and oil always separate. However, there will likely be a change in appearance and physical properties of the oil in tube four because it will be mixed with whatever the microbial solution comprises. When I first set up the experiment and let the tube components settle, with tube three, the liquid layer was clear, and there was a thick, flat layer of oil on top with no bubbles. When tipped horizontally, the oil stayed together as a thick layer over the liquid layer and moved slowly from side to side. When I put it back upright in the rack, the oil layer seemed to coagulate to one side of the tube and turned cloudy. The top of the oil layer looked concave.

9 Initially, tube four's liquid layer was cloudy and yellowish, with a thick, cloudy oil layer on top. When tipped horizontally, the oil layer moved over the liquid layer in what looked like individual bubbles. It moved freely and fast from side to side and looked slightly bouncy. When I put it back upright in the rack, the oil layer went right back up to the top and remained a bubbly, thick layer. For tube three on day one of observations, the liquid layer was clear, and the oil layer was still coagulated on one side of the tube and was cloudy. The top still looked concave. When tipped horizontally, the oil stayed together as a thick layer, with some individual bubbles present within the thick layer. The oil still moved slowly from side to side and looked bouncy. The liquid layer stayed clear. When I put it upright back in the rack, the oil coagulated back to the side of the tube, and instead of being cloudy, it was mostly clear. The top of the oil layer remained concave. For tube four, a whiteish substance sank to the bottom of the tube. The liquid layer was still cloudy but looked a little more yellow. The oil layer was still bubbled, however, it looked like it was two different colors; the original color of the oil and a brownish-orange hue. When tipped horizontally, the whiteish substance mixed with the liquid layer made it extremely cloudy. The oil layer was still composed of individual oil bubbles, that did not conjoin when moved from side to side, and the oil moved fast. When I put it back upright in the tube, the oil went back up to the top as a thick, bubbly layer and still looked like it had two different colors, as previously mentioned in the initial observation day. The liquid was way more cloudy, and the substance at the bottom of the tube was no longer present, as it mixed with the liquid layer. For tube three on day two of observations, the liquid layer was still clear and the oil was still coagulated to one side of the tube and was cloudy. The top of the oil layer still looked concave. When tipped horizontally, the oil stayed together as a

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10 thick layer over the liquid layer, with some individual oil bubbles in the thick layer. The oil moved slowly from side to side and looked bouncy. When I put it upright back in the rack, the oil coagulated back to the side of the tube and was still clear. The top of the oil layer also continued to look concave. For tube four, the whiteish substance settled back to the bottom of the tube. The liquid layer was still cloudy and was still yellow in color. The oil layer was still thick and bubbly, and it still looked like it was two different colors; the original color of the oil and a brownish-orange hue. When tipped horizontally, the whiteish substance mixed with the liquid layer and made it more cloudy again. The oil layer was still composed of individual oil bubbles that did not conjoin when they hit each other, and the oil still moved fast. When I put it back upright in the rack, the oil went back up to the top as a thick, bubbly layer, and it still looked like it was the two different colors from the previous days. The liquid became more cloudy, and the whiteish substance at the bottom of the tube was no longer present because it mixed with the liquid layer. For tube three, on day three of observations, the liquid layer was still clear, and the oil was still coagulated to one side of the tube and looked cloudy. The top of the oil layer still looked concave. When tipped horizontally, the oil layer stayed together as a thick layer over the liquid layer, and individual oil bubbles were still present within the thick layer. The oil still moved slowly from side to side and looked bouncy. When I put it back upright in the rack, the oil layer coagulated back to the side of the tube. However, this time it looked clear. The top of the oil layer still looked concave. The liquid layer remained clear, as well. For tube four, a whiteish-yellow substance settled at the bottom of the tube. The oil layer consisted of one large individual oil bubble and some very small individual bubbles. The liquid layer was clear, but yellowish in color. When tipped horizontally, the

11 whiteish-yellow substance mixed with the liquid layer, making it more cloudy and yellow. There seemed to be very little oil, as there were only a few individual oil bubbles compared to previous days. When I put it back upright in the rack, the oil layer remained bubbly; however, the size of the oil bubbles was now medium and small. The liquid layer was more cloudy and more yellow in color. Given the results of this experiment and the observations I made along the way, I accept my hypothesis that there is no change in the appearance and physical properties of the oil in tube three. Still, there being a change in the appearance and physical properties of the oil in tube four as being correct. For tubes five and six, the environmental factor I chose to alter was the temperature. Tube six with the hot water will experience a greater oil breakdown than tube five with the cold water. When I first set up the experiment and let the tube components settle, the liquid layer was cloudy and yellowish in color for tube five. The oil layer was thick and bubbly. When tipped horizontally, the oil layer presented itself as individual oil bubbles. It moved fast when moved from side to side, and looked bouncy. When I put it back upright in the rack, the oil returned to the top of the liquid and stayed a bubbly, thick layer. The liquid layer remained cloudy and yellowish in color. For tube six, the liquid layer was cloudy and yellowish in color. The oil layer was thick and bubbly. When tipped horizontally, the oil layer presented itself as individual oil bubbles. It moved fast and freely and looked bouncy from side to side. When I put it back upright in the rack, the oil layer returned to the top of the liquid layer and stayed a bubbly, thick layer. The liquid layer remained cloudy and yellowish in color. For tube five, during day one of observation, a pink substance settled at the bottom of the tube. The liquid layer was clear, but yellow in color. The oil layer

12 was thick and bubbly; however, the bubbles were medium to large, unlike the other tubes with bubbly layers. When tipped horizontally, the pink substance that was at the bottom of the tube mixed with the liquid layer and made it more cloudy and more pink in color. The oil showed larger individual bubbles that did not conjoin when moved from side to side. It moved fast and looked bouncy. When I put it back upright in the rack, the pink substance at the bottom of the tube was no longer present as it mixed with the liquid layer. The liquid layer was now very cloudy and yellowish-pink in color. The oil layer was still thick and bubbly, and the bubbles were medium to small in size. The oil layer also looked like it was two different colors; the original color of the oil and a tannish hue. For tube five, during day two of observations, everything was the same as day one. For tube six, everything was the same as day one as well. For tube five, a pink substance settled at the bottom during day three of observations. The oil layer was thick and flat; it had no bubbles and looked concave. The liquid layer was clear and yellow in color. When tipped horizontally, the pink substance mixed with the liquid layer, making it more cloudy and pink. There seemed to be very little oil, as there were only a few individual oil bubbles compared to previous days. When I put it back upright in the rack, the oil layer showed as bubbly with medium-sized bubbles. The liquid later was more cloudy and was now a pinkish-yellow color. For tube six, a pink substance settled at the bottom of the tube. The oil layer was thick and flat, had no bubbles, and looked concave. The liquid layer was clear and yellow in color. When tipped horizontally, the pink substance mixed with the liquid layer, making it more cloudy and pink. There seemed to be very little oil, as there were only a few individual oil bubbles compared to previous days. When I put it back upright in the rack, the oil layer

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13 showed as bubbly with medium-sized bubbles. The liquid layer was more cloudy and pinkish-yellow in color. Given the results of this experiment and the observations I made along the way, I deny my hypothesis of tub six experiencing a more significant breakdown of oil than tube five because of the hotter water temperature I chose to use. Both tubes had the same amount of oil breakdown regardless of the temperature, so my hypothesis was proven incorrect. Discussion In this lab, I have learned the importance of monitoring large oil spills and other aspects that oil may end up in water. When oil spills occur, food supplies are contaminated, animals can be gravely harmed, and they may end up dying as a result, and the environment is also heavily affected. Oil spills play a part in global warming, so it is imperative that we need to put forth time, effort, and resources into renewable resources. I did not run into any obstacles during this lab; besides that, I did not account for this lab taking multiple days to complete. References Venosa, A. (n.d.). Nrt fact sheet: Bioremediation in oil spill response.

SCI207 Week 5 Lab Report - Kimberly Selby

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