Name: _____________________________ Date: __________________________

SBI3C Microbiology Lab 1:​ ​Bacteria Near and Dear

Background
Like all organisms, bacteria thrive on optimal conditions such as constant temperature and
a standard food source. In this activity, you will capture and culture bacteria on a
nutrient-rich agar medium. The petri dish containing the medium has been sterilized to
remove all contaminants. As you work, be careful to maintain sterile (aseptic) conditions. In
this confirmation lab, you will discover that bacteria are everywhere.

Learning Goals
■ I can identify the conditions that bacteria need to grow and survive
■ I can identify physical characteristics of bacteria
■ I can demonstrate aseptic conditions and swabbing culturing techniques
■ I can describe and characterize bacterial growth
■ I can describe how to practice good hygiene and explain its importance

In this lab you will swab culture medium to observe and describe the characteristic of
bacterial growth.

Pre-Lab Component

Lab Safety
1. Treat all microorganisms as potential pathogens.​ Under unusual circumstances a
few microorganisms that are not normally pathogenic can act as pathogens.
2. Sterilize equipment and materials. ​All materials, media, tubes, plates, and other items
used for culturing microorganisms should be sterilized.
3. Disinfect work areas before and after use. ​Use a disinfectant. Either alcohol or bleach
can be dangerous if splashed in the eyes. Students should know where the nearest
eyewash station and sink are located.
4. Wash your hands.​ Use a disinfectant soap to wash your hands before and after working
with microorganisms. Gloves may be worn as extra protection.
5. Do not eat or drink in the lab, nor store food in areas where microorganisms are
stored.​ Never eat or drink in the laboratory. Keep your fingers out of your mouth. Cover
any cuts on your hands with a bandage. Gloves may be worn as extra protection.
6. Label everything clearly. ​All cultures, chemicals, disinfectant, and media should be
clearly and securely labeled with their names and dates.
7. Dispose all waste material correctly. ​All items to be discarded after a class, such as
culture tubes, culture plates, swabs, toothpicks, wipes, and gloves, should be placed in a
biohazard bag.

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Name: _____________________________ Date: __________________________

Pre-lab questions
1. What is a pathogen? Are pathogens harmful or helpful? ​(1 K)

2. Do you think bacteria are harmful or helpful? Why? ​(2 C)

3. What are some places we can find bacteria? List at least 2-3 places. ​(2 K)
●

4. Why are we not allowed to swab certain surfaces within the school? ​(1 T)

5. How might we remove bacteria from various surfaces? ​(2 K)

6. Complete the table below with corresponding definitions and lab components. ​(4 K)

Term Definition In the Lab

Control

Independent
Variable

Dependent Variable

Control Variable

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Name: _____________________________ Date: __________________________

Day 1 - Lab

Hypothesis:
Where will you find the most bacteria? Why do you think so? (2 A)
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________

Will you see bacteria on the mouth bacteria plate? Why or why not? (1 A)
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________

Materials
1. Gloves 6. Hand lens
2. Sharpie marker 7. Blue tray
3. Prepared disposable petri dishes 8. Masking tape
4. Cotton swabs 9. Three sources of bacteria
5. Disinfectant

Methods
Part A: Instructions for plating the mouth bacteria (individually)​:
Before you begin, make sure you have completed the pre-lab,
listed your locations and written a hypothesis.
1. Wipe down the counter with disinfectant.
2. Obtain one petri dish and mark four quadrants on the ​bottom
as shown on the right, with a sharpie.

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Name: _____________________________ Date: __________________________
​ n the ​side​ of the petri dishes along with the ​date.​
3. Mark your ​initials o
4. Do not plate any bacteria on quadrant 1. This will be your ​control​.
5. Lick your lips and swab them with a sterile swab.
6. Brush the swab lightly on the sterile agar in the petri dish. Swab quadrant 2, then
3 and then 4. ​You will only swab your lips once
7. Place lid on petri dish and tape it shut across the top and bottom
to form an X.

Part B: Instructions for plating the school bacteria (group)​:

Person A
9. Obtain a sample of bacteria from a known location. ​Record this location​ and
plate the bacteria on the agar in quadrant 2.
Person B
10. Obtain a sample of bacteria from a known location. ​Record this location​ and
plate the bacteria on the agar in quadrant 3.
Person C
11. Obtain a sample of bacteria from a known location. ​Record this location​ and plate
the bacteria on the agar in quadrant 4.
12. Place lid on petri dish and​ tape it shut across the top and bottom ​to form an X.
13. Incubate the cell culture ​upside down​ at 30 degree Celsius for 48 hours.

Person Name Location

A (2)

B (3)

C (4)

Teacher Initial ​__________________

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Name: _____________________________ Date: __________________________

Colony Morphology: Describing Bacterial Colonies

Form​: Shape of the ​entire​ colony.

Size: ​Measure the diameter (mm) of a colony.

Surface: ​Bacterial colonies are frequently shiny and smooth in appearance. Other
surface descriptions might be: ​veined, rough, dull, wrinkled (or shriveled),
glistening​.

Texture: ​Several terms that may be appropriate for describing the texture or
consistency of bacterial growth are: ​dry, moist, mucoid, brittle, viscous, butyrous
(buttery)​.

Colour: ​Describe the color or pigment of the colony. Also include descriptive terms for
any other relevant optical characteristics such as: ​opaque, cloudy, translucent,
iridescent​.

Elevation: ​This describes the “side view” of a colony as it rises above the agar. These
are the most common.

Margin: ​The margin or edge of a colony (or any growth) may be an important
characteristic in identifying an organism. Several examples are shown below.

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Name: _____________________________ Date: __________________________

Day 2 - Lab
1. Disinfect your bench and obtain your petri dishes from the front.
2. Using a hand lens, observe the colonies and complete the observation table. ​Do
not open the petri dishes. There is a helpful guide on the last page for you.
3. Once you are done, ​hand in your petri dishes for disposal.

Compare the mouth bacteria dishes with others in your class. You will need this
information for the discussion questions.

Results
Observations - **Use the information on Page 5 to complete the table below**

Table 1: __________________________________________________________ (2 C)
Colony Morphology (from agar plates)
Colony # of
Size Colour Surface Texture
Location Colonies Form Elevation Margin
(mm)

Control

Mouth
Bacteria

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Name: _____________________________ Date: __________________________

Draw one colony from each location. Label the locations in the diagram and
provide a caption for what you observed. (4C)

Figure 1: ______________________________________________________________
______________________________________________________________________
______________________________________________________________________

Post-Lab Discussion: Are Bacteria Good or Bad? Practicing Good Hygiene

2. Compare the growth on your mouth dish with others in the class. What factors
contribute to differences? Why would there be similarities?​ (2 T)

3. If colonies appeared on your control quadrant, what conclusions can you make
about your experiment? ​(2 T)

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Name: _____________________________ Date: __________________________

4. Under favorable conditions, bacteria may reproduce once every 20 minutes. At this
rate, how many bacteria could be produced from a single bacterium in 6 hours,
assuming all survive? Show your calculations. ​(3 A)

5. Why did we not open the petri dishes when observing the bacteria?​ (2 T)

6. What are some other considerations we should make when doing this lab? ​(2 A)

7. What would you do differently if you were to do the lab again? ​(2 C)

8. Why are we not allowed to bring the lab plates home? ​(2 A)

9. List three ways in which we can practice good personal hygiene?​ (3 A)