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Old 02-19-2006, 10:43 PM
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Wink That's Weird!

That's weird! It worked before. Anyways, I copied it and lets hope it'll fit on here (it's pretty long.) Good luck!


Question: Will pollution affect bioluminescent bacteria?

Hypothesis: Yes, I think that pollution does affect bioluminescent bacteria, because this will disrupt the entire ecosystem.

1. 5-10 deceased salt-water fish
2. salt water (for the fish)
3. a refrigerator (4 - 6C)
4. South Saskatchewan River water
5. Tap water
6. Liquid soap
7. Oil
8. Metal transferring loop
9. 16-26 Petri dishes
10. syringe
11. light-meter or thin translucent paper
12. tape
13. sharpie
14. syringe
15. Bunsen burner
16. Denatured alcohol
17. Lighter
18. Rubbing alcohol
19. Bleach
20. 30g NaCl
21. 1g Glycerol
22. 10g Peptone (Bacto-Peptone)
23. 3g Beef extract
24. 15g of Bacto-Agar
25. Water with a ph of about 7.3

1. Gather materials
2. Place fish half submerged in salt water
3. Refrigerate at 4 6C
4. Check each day for bioluminescent bacteria growth
5. Clean Petri dishes with bleach and water
6. Dry Petri dishes, allowing no air to enter
7. Mix 30g of NaCl, 1g glycerol, 10g peptone (bacto-peptone), 3g beef extract, and 15g of bacto-agar
8. Dilute the ingredients in water with a ph of about 7.3
9. Fill with water up until 1000mL
10. Separate solution (BOSS) evenly into Petri dishes
11. Sterilize working area with rubbing alcohol
12. Fill bottom of the Bunsen burner with denatured alcohol
13. Light the wick with a lighter
14. Hold the metal transferring loop in the flame until the metal turns red.
15. Cool the loop on one of your agar plates
16. Using the loop, scrape off some bacteria, and place it on a separate agar plate
17. Close the lid to your agar plate
18. Place tape across the top of your plate
19. Label this plate, with a sharpie, River water #1
20. Repeat steps 14-19, 3-5 times, depending on the amount of Petri dishes you have
21. For each time you repeat step 19, use a higher number (river water #2)
22. Repeat steps 14-21, 3-5 times, replacing the words river water with tap water
23. Repeat steps 14-21, 3-5 times, replacing the words tap water with liquid soap
24. Repeat steps 14-21, 3-5 times, replacing the words soap with oil
25. Dispose of the agar that was used to cool the metal transferring loop
26. Clean the Petri dish that contained this agar
27. Place all other agar plates in a dark room (at a constant temperature of 18C)
28. Using the light-meter, measure the luminosity of each dish
29. Record data
30. Measure 3-5 (depending on the amount of Petri dishes you have) even amounts of each pollutant with a sterile syringe
31. Squirt the pollutant onto the luminescent bacteria that is labelled the same
32. Close the lid after applying the pollutant
33. Observe results 1 hour after application, 1 day after application, 2 days after application, 3 days after application, 4 days after application, 5 days after application, 6 days after application, and 7 days after application
34. Record results
35. Clean and replace all materials

Application: You could use bioluminescent bacteria to measure the degree of water pollution.

Extension: I could extend this project by using different pollutants, varying the amounts of these pollutants, repeating this experiment more times, create a liquid culture, changing the temperature of the pollutants, changing the temperature of the bacteria, varying the amounts of the bacteria, using different bacteria, or changing the ph of the pollutants.

Research topics for background information:
Effects of pollutants
Effects of pollutants on water
Effects of pollutants on bacteria
Marine ecology
River water
South Saskatchewan River water
Tap water
Bacteria cultures

Web sites:

Joker Jones
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