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Third method

Reason for research

  • 7 tubes
  • The first tube is a control. In this tube, BHB, Bushnell Hass broth, is alone.
  • The second tube consists of BHB plus the bacteria, pseudomonas putida.
  • The third tube, also a control, consists of BHB plus the Styrofoam.
  • The fourth tube, and the final control, contains BHB plus oil.
  • Next is the fifth tube, the first of the experimental tubes. In this tube, there is BHB, Styrofoam, and the bacteria.
  • The sixth tube contains BHB, the bacteria, and also now oily Styrofoam.
  • In the seventh and final tube I put BHB, oil, and the bacteria into the tube.

Method 2 (continued)

  • I first experimented with a glass slide instead of Styrofoam to see if the a biofulm would actually grow
  • Once the proper steps were taken to stain the glass, I would then put the slide under a microscope and see if a biofilm has actually grown onto the Slide.
  • I can tell if it has grown if I see the pink pseudomonas putida clustered together.
  • This method did not work out because we couldn't find a way to determine if the bacteria are growing or if even if there was a biofilm on the styrofoama

Polystyrene (Styrofoam)

  • Pollution
  • Most of the world's pollution is made of petroleum based products i.e. Styrofoam and Plastics
  • Other methods used today to deal with this pollution are harmful to our environment
  • Polystyrene is a petroleum-based plastic made from the styrene monomer
  • Polystyrene is a light-weight material, about 95% air, with very good insulation properties
  • The process of creating polystyrene is called polymerization.
  • Main component is petroleum

Hypothesis

If the Pseudomonas biofilm is grown in a liquid growth medium that lacks a Carbon source, the bacteria will turn to the styrofoam as a carbon source and begin to degrade it

Degradation of Styrofoam using Pseudomonas Putida and Pseudomonas Aeruginosa

By,

Jay Patel

Pseudomonas Aeruginosa

Pseudomonas Putida

  • The microbe is free-living, usually found in moist environments like soil, marshes, and water, like the other members of the Pseudomonas genus
  • Pseudomonas aeruginosa is a common gram-negative, rod-shaped bacterium that can cause disease in plants and animals, including humans.
  • Pathogen
  • Can be used in bio remediation.
  • Pseudomonas putida is a versatile environmental isolate that is capable of growth on several aromatic hydrocarbons, including benzene (C6C6) and toluene (C7C8).
  • Benzene is a natural constituent of crude oil, and is an organic chemical compound with the molecular formula C6C6.
  • hydrocarbon.
  • Toluene is formerly known as tolul, is a clear, water-insoluble liquid with the typical smell of paint thinners.
  • Has a unique and diverse metabolism that makes it possible to biodegrade oil

Spectrophotometer

Conclusion

  • I would like to conclude by pointing out that bacteria was growing in the tube that contained styrofoam BHB and bacteria.
  • This shows promise for the future
  • This project requires more work for it to be completed in the future
  • If I had more time, I would've been able to do more with my last experiment

Petroleum

  • Started this project at the end of my sophomore year
  • Measures the absorption of each tube to determine if the bacteria are proliferating or not.
  • The first detail I would like to point out the significant difference of transmittance between the third control, BHB and Styrofoam, and the fifth tube, which has BHB, Styrofoam and the bacteria.
  • The transmittance between the two is differentiated by almost fifty.
  • Clearly shows that there is activity in this tube; otherwise the transmittance wouldn’t be this low.
  • The transmittance is also low for the sixth and seventh tubes. They both have their transmittance around the forty marks which also indicates that there is activity within both tubes like the fifth tube.

Importance

First Method

Bacteria

  • The first method that I thought of required a test tube, broth, Pseudomonas Putida, and Styrofoam.
  • The Bushnell Broth was used in my experiment due to its lack of nutrients the bacteria need to survive.
  • This broth did not contain hydrocarbon
  • I would first place the broth into a test tube.
  • Then I would follow and put the Styrofoam into the test tube.
  • But then we would drop 1mL of bacteria onto the Styrofoam before it is put into to the tube of broth.
  • With this experiment, the only food that is available for the bacteria is the Styrofoam.
  • Since there is no food found in the broth for the bacteria, they will be forced to eat the Styrofoam.
  • naturally occurring, yellow-to-black consisting of a complex mixture of hydrocarbons of various molecular weights and other liquid organic compounds that are found in geological formations beneath the Earth's surface.
  • Not as thick as oil
  • The main component of several plastics and Styrofoam
  • In this experiment I can tell if the Styrofoam is being consumed if there is a growing air bubble on the top of the tube.
  • This bubble would be of the metabolic gases the bacteria would release when eating the Styrofoam
  • There was an air bubble, but after weeks increasing, the bubble would stop and become stagnant
  • pollution is destroying our environment and the organisms that inhabit it.
  • There is not efficient way to degrade this type of pollution
  • Burning causes air pollution
  • There is no room
  • Pseudomonas Putida
  • Pseudomonas Aeurginosa

Deep water Horizon oil spill

  • Was caused by a wellhead blowout
  • 210 million gallons of oil had reached the surface of the war and was pushed all throughout the world through ocean currents
  • Cleaning this took a lot of manual labor but there were also biological processes which can be as effective or even more
  • A process includes having bacteria eat the oil and dispose of the several toxic chemicals
  • Bacteria that can be used in bio remediation, which is a waster management technique that allows organic structures to remove or break down pollutants from contaminated sites.

Second Method

How to create a biofilm

1. Select one of the square petri plates containing the broth.

2. Onto the surface of the agar, aseptically place one sheet of sterile filter paper. Sterilize a pair of forceps by dipping them in a container of alcohol and flaming them.Place the sheet so that no air bubbles are trapped under the filter paper.

3. Dilute the overnight culture. Thoroughly mix the culture by vortexing or by rolling the tube containing the culture between the palms of your hands.

4. Uniformly moisten the filter paper with 1 ml of the diluted bacterial culture.

5. Select a clean 1 X 3 inch microscope slide and using the forceps, immerse the slide in a container of alcohol and sterilize it by flaming. Then carefully lay the slide on top of the filter paper. make sure there are no air bubbles

6. Repeat step 5 three more times.

7. Between Periods 1 and 2 If possible, remoisten the plate after 24 hours with 1 ml of the culture.

Period 1

1. After two days the biofilms may be harvested by carefully lifting the slides from the filter paper surface with a pair of sterile forceps (flamed). The biofilm should appear as a slimy layer coating the under surface of the slide. Lifting too rapidly may disturb the biofilm causing large sections to slough off the slide.

  • This method revolved around creating biofilm
  • A biofilm is any group of microorganisms in which cells stick to each other and often these cells adhere to a surface.
  • . If the conditions are right, the bacteria grow and reproduce, creating a thick, slimy surface.
  • The bio film was meant for an easier way for the bacterium to eat the Styrofoam.
  • In this method we would first need to clean the styrofoam with the use of an incubator
  • Then you would use the following steps to make cause a bio film to grown onto the styrofoam
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