Bioremediation help
Posted: Sat Nov 14, 2015 6:23 pm
Hey everyone! I'm a high school freshman, and I desperately need some help! I want to test whether the pH of the water will affect the rate at which bioremediation occurs. I originally planned on using Alcanivorax borkumensis, but the only place that I found sells it is atcc, which sells it for $300. Unfortunately, my family currently does not have that money to spend on a school project. My teacher told me I can a) find a cheaper substitute for both the bacteria and the crude oil, b) find a cheaper substitute bacteria that can also break down oil, or c) change my project completely. Obviously, option c would be least desirable. Ideally, option b would be best, or if anyone knows where I can find Alcanivorax borkumensis much cheaper, that'd be fantastic! Here is my current procedure if this helps, and if anyone has any tips for changes that'd be great. Any help will be appreciated! Thanks!
Culturing the bacteria
Using a pipet, withdraw 0.5 milliliters of marine broth from a container with 5 milliliters of marine broth. Use the broth in the pipet to rehydrate the pellet of Alcanivorax borkumensis and transfer this mixture to the original 5 mL of marine broth to revive the bacteria from its freeze-dried state. Mix well. Use a pipet to transfer the suspension to the agar plate, to allow the bacteria to culture. Incubate the plate aerobically at 30°C for 24 hours.
Preparing the cups
The next steps will be used to mimic the conditions of saltwater found in the ocean, and prepare each cup for introduction of oil and Alcanivorax borkumensis. Pour all of the salt into all
of the water. Mix well until all the salt is completely dissolved. Label the cups “8.3(1)”, “8.3(2)”, “8.3(3)”, “8.1(1)”, “8.1(2)”, “8.1(3)”, “7.9(1)”, “7.9(2)”, “7.9(3)”, “Control 1”, “Control 2”, and “Control 3”. Measure the pH of the water to determine how much pH change is necessary. Fill each cup with 200 milliliters of the salt water mixture. Add basic solution or acidic solution as needed to each cup to match the pH level as labeled on each cup. Do not change the pH of the control cups. Once all the cups have the correct pH, add 20 milliliters of oil into each cup, then add 5 colonies of Alcanivorax borkumensis into each cup from the agar plate using an inoculating loop. Store all cups in 23°C for 24 hours.
Data Collection
The next steps will be used to determine how much oil has been degraded in each cup since the previous day. Pour the solution in “Control 1” into the separatory funnel, and funnel the water back into the cup. Keep the oil in the funnel. Pour the oil into the graduated cylinder. Record the volume of oil. Pour the oil back into the original cup to allow bacteria to continue bioremediation. Repeat these steps for each cup. Repeat these steps over the next three days.
Culturing the bacteria
Using a pipet, withdraw 0.5 milliliters of marine broth from a container with 5 milliliters of marine broth. Use the broth in the pipet to rehydrate the pellet of Alcanivorax borkumensis and transfer this mixture to the original 5 mL of marine broth to revive the bacteria from its freeze-dried state. Mix well. Use a pipet to transfer the suspension to the agar plate, to allow the bacteria to culture. Incubate the plate aerobically at 30°C for 24 hours.
Preparing the cups
The next steps will be used to mimic the conditions of saltwater found in the ocean, and prepare each cup for introduction of oil and Alcanivorax borkumensis. Pour all of the salt into all
of the water. Mix well until all the salt is completely dissolved. Label the cups “8.3(1)”, “8.3(2)”, “8.3(3)”, “8.1(1)”, “8.1(2)”, “8.1(3)”, “7.9(1)”, “7.9(2)”, “7.9(3)”, “Control 1”, “Control 2”, and “Control 3”. Measure the pH of the water to determine how much pH change is necessary. Fill each cup with 200 milliliters of the salt water mixture. Add basic solution or acidic solution as needed to each cup to match the pH level as labeled on each cup. Do not change the pH of the control cups. Once all the cups have the correct pH, add 20 milliliters of oil into each cup, then add 5 colonies of Alcanivorax borkumensis into each cup from the agar plate using an inoculating loop. Store all cups in 23°C for 24 hours.
Data Collection
The next steps will be used to determine how much oil has been degraded in each cup since the previous day. Pour the solution in “Control 1” into the separatory funnel, and funnel the water back into the cup. Keep the oil in the funnel. Pour the oil into the graduated cylinder. Record the volume of oil. Pour the oil back into the original cup to allow bacteria to continue bioremediation. Repeat these steps for each cup. Repeat these steps over the next three days.