11.4: Materials and Procedures
- Page ID
- 40221
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Temperature growth ranges
- Student stock TSB cultures-
- Pseudomonas fluorescens
- Bacillus cereus
- Escherichia coli
- Instructor provided TSB culture-
- Streptococcus mutans
- 16- TSB tubes
- Prodigiosin production
- Student stock TSA plate culture-
- Serratia marcescens
- 2- TSA Plates
- 4-Sterile transfer pipettes
- Buckets labeled according to incubation temperature
Procedures (2 Groups per Table)
Temperature growth ranges
- Obtain the 16 test tubes of TSB and label the tubes with the organism and the temperature. Each organism will be inoculated into 4 tubes of TSB. These four tubes will then be inoculated at 4 different temperatures.
- So, there will be 4 test tubes for each of the 4 organisms:
P. fluorescens → 5ºC, 25ºC, 37ºC, 42ºC
B. cereus → 5ºC, 25ºC, 37ºC, 42ºC
E. coli → 5ºC, 25ºC, 37ºC, 42ºC
S. mutans → 5ºC, 25ºC, 37ºC, 42ºC
- Using a sterile transfer pipette inoculate 1 drop of the organism into the 4 separate tubes of TSB. Place the correctly labeled and inoculated test tube of each organism into each temperature bucket on the front table:
5ºC
25ºC
37ºC
42ºC
Prodigiosin production
1. Streak 2 TSA plates with Serratia marcescens. Label one for incubation at 25ºC and one for incubation at 42ºC.
2. Place, inverted, on the correct side (25ºC or 42ºC) of the pan labeled for incubation.
Results:
Temperature growth ranges
- After incubation, retrieve your group’s TSB tubes from the labeled buckets and mix them gently and evenly distribute the bacteria in the tube.
- Compare all the tubes of the same species to each other (all the E. coli to each other, etc.). Score them according to how turbid the tubes are and record the results in the table:
0 = no growth (clear)
1 = slightly turbid
2 = moderately turbid
3 = very turbid
(Note that you can also compare all the species to each other at a given temperature.)
Enter your ratings for each bacterium/temperature and give the temperature classification for bacterium.
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Organism |
5ºC |
25ºC |
37ºC |
42ºC |
Optimum Temperature |
Temperature Classification |
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Prodigiosin production
- Retrieve the Serratia marcescens TSA plates. Observe and compare the differences in growth at the two temperatures, 25ºC and 42ºC.
- Record your observations in the table.
Enter your observations on the growth characteristics of Serratia marcescens
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Temperature |
Growth Characteristics |
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25ºC |
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42ºC |
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Contributors and Attributions
Kelly C. Burke (College of the Canyons)