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Copyright Glencoe/McGraw-Hill, a division of the McGraw-Hill Companies, Inc
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Density indicator strips are used to monitor the rate of microbial growth These strips are attached to the culture vessel As the microbes multiply, the solution becomes cloudy, obscuring some of the shaded strips The degree of visibility of the shaded strips indicates the density of the microbes In this activity, you will observe the bioremediation effectiveness of the fungus Penicillium and the bacteria Pseudomonas on a sample of oil
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How effective are Penicillium and Pseudomonas for breaking down oil
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Observe the effect of hydrocarbon-degrading microbes on oil Observe microbes degrade oil
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Penicillium sp culture Pseudomonas sp culture sterilizing, disinfectant solution lightweight oil nutrient fertilizer density indicator strips (5) paper towels
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30-mL sterile culture test tubes with screw caps (3) 250-mL beaker sterile 25-mL graduated cylinder plastic dropping pipettes (4) labels (4) sterile distilled water 201
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Chemistry: Matter and Change 26
Name
Date
Class
LAB 261
LABORATORY MANUAL
Safety Precautions
Always wear safety goggles, a lab apron, and gloves Dispose of wastes as directed by your teacher Organisms or living materials should always be treated and handled as if they were hazardous Observe proper personal hygiene when handling microorganisms Be sure to wear gloves and wash your hands with antibacterial soap or detergent after removing the gloves
Pre-Lab
1 Briefly explain bioremediation and
biostimulation 2 Explain why it is necessary to disinfect the work area 3 Explain the function of the density indicator strips 4 Read the entire laboratory activity Form a hypothesis about the effect the microbes will have on the oil suspended in the water Record your answer in the next column
Procedure
1 Use the disinfectant solution according to the 10
4 5 6
manufacturer s directions to thoroughly clean and disinfect your work area Wipe the area with paper towels and dispose of the towels as directed by your teacher Wash your hands with antibacterial soap Affix labels to each of the sterile culture test tubes Label them all with your name Label one test tube Penicillium, the second test tube Pseudomonas, and the third test tube control Place a label with your name, date, and class period on the 250-mL beaker Using a 25-mL graduated cylinder, pour 15 mL of distilled water into each sterile culture test tube Add 12 drops of oil to each test tube Add about 01 mL of nutrient fertilizer to each test tube The control test tube should not receive any additional materials Being careful not to pick up other microbes by laying down equipment, add about 3 mL of Penicillium culture to the appropriate test tube Add about 3 mL of
Chemistry: Matter and Change 26
Hypothesis
Cleanup and Disposal
1 Dispose of materials as instructed by your
teacher 2 Return all lab equipment to its proper place 3 Using the disinfectant solution, disinfect your work area 4 Wash your hands thoroughly with antibacterial soap before leaving the lab
Laboratory Manual
Copyright Glencoe/McGraw-Hill, a division of the McGraw-Hill Companies, Inc
Pseudomonas culture to the Pseudomonas test tube Securely fasten the top on each test tube Shake each tube gently to ensure thorough mixing of the contents Number the density strips from 1 to 5, with 1 being the lightest coloration Secure the strips to the outside of each test tube so that the density strip is visible through the solution The top set of bars should be just below the level of the liquid in the test tube Record the visibility of the density strips, the color of the liquid, and the general appearance of the contents of the test tube on Data Table 1 Loosen the caps on the test tubes about half way and place them all in the labeled 250-mL beaker Place the specimens in a warm location in the classroom CAUTION: Do not touch bacteria cultures Repeat the same observations daily and record your observations on Data Table 1 for a total of 5 days
Name
Date
Class
LAB 261
LABORATORY MANUAL
Data and Observations
Data Table 1
Day 1 Test-tube contents control Density strip reading Color of liquid General appearance of test-tube contents
Penicillium
Pseudomonas
control
Penicillium
Pseudomonas
control
Copyright Glencoe/McGraw-Hill, a division of the McGraw-Hill Companies, Inc
Penicillium
Pseudomonas
control
Penicillium
Pseudomonas
control
Penicillium
Pseudomonas
Laboratory Manual
Chemistry: Matter and Change 26
Name
Date
Class
LAB 261
LABORATORY MANUAL
Analyze and Conclude
1 Observing and Inferring What changes occurred in each test tube as the 5 days
progressed
2 Observing and Inferring Did one organism break down the oil better than the other organism
3 Observing and Inferring What happened to the cloudiness of the tubes as the 5 days
progressed
4 Acquiring and Analyzing Information What does an increase in the cloudiness of the
system indicate
5 Acquiring and Analyzing Information What did the changes in the color of the
system and the general appearance indicate
6 Designing an Experiment/Identifying Variables What was the function of the
Copyright Glencoe/McGraw-Hill, a division of the McGraw-Hill Companies, Inc
control test tube
7 Designing an Experiment/Identifying Variables What was the purpose of shaking
the test tubes and then leaving the caps partially opened
8 Error Analysis Compare your results to those of other students in the class What could
be the cause of some differences
Real-World Chemistry
1 What effect might the use of microorganisms 2 Why might knowledge of pH be useful when
that are not native to a site have on the ecology
using bioremediation techniques
Chemistry: Matter and Change 26
Laboratory Manual
Name
Date
Class
LABORATORY MANUAL
Use with Section 262
Growth of Algae as a Function of Nitrogen Concentration
reshwater comes from many sources, including lakes, rivers, and municipal reservoirs However, daily activities of life often leave these water sources polluted and unfit for personal consumption or use in industry Nitrogen and phosphorus compounds are among the most common pollutants They contribute to pollution by causing algae and bacteria in the water to reproduce rapidly When these organisms die, the decomposition process depletes oxygen in the water, killing fish and other aquatic life In this lab, you will investigate the effect of nitrates on algae
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