Bacteriology (77 pts)

NAME _________________________________________________________ (1 pt)

TA NAME _______________________ (1pt) SECTION#_________ (1 pt)
Read the Introduction
Are bacteria prokaryotic or eukaryotic? (1 pt)

What are three types of energy sources for prokaryotes? (3 pts)

What is the main component in the bacterial cell wall? (1 pt)

How is DNA organized in a prokaryote? (1 pt)

How do prokaryotes reproduce? (1 pt)

Morphology of Individual Cells

Using the provided slides of mixed bacteria, state the name and three major shapes of
bacteria. (3 pts)
*Hint: you need to use 100X oil immersion to see these shapes clearly. Be careful with
the immersion oil and use only what is necessary.

Name Description of Shape

Identifying Bacteria by the Gram Stain Procedure
Carefully heat fix bacteria to a slide from an agar plate.
1. What is the first stain in a Gram Stain procedure? (1 pt)
What is the purpose of the first stain? (1 pt)
If you stopped here, how would the bacteria be colored? (2 pt)

Stain Purpose Gram Positive Gram Negative

2. What is the second stain in a Gram Stain procedure? (1 pt)

What is the purpose of the second stain? (1 pt)
If you stopped here, how would the bacteria be colored? (2 pt)

Stain Purpose Gram Positive Gram Negative

3. What is the third stain in a Gram Stain procedure? (1 pt)

What is the purpose of the third stain? (1 pt)
If you stopped here, how would the bacteria be colored? (2 pt)

Stain Purpose Gram Positive Gram Negative

4. What is the fourth stain in a Gram Stain procedure? (1 pt)

What is the purpose of the fourth stain? (1 pt)
If you stopped here, how would the bacteria be colored? (2 pt)

Stain Purpose Gram Positive Gram Negative

Share class results and complete the chart (4 pts)

Name of Bacteria Results of Gram Stain

Escherichia coli

Serratia marcescens

Staphylococcus aureus
 Bacillus subtilis

Exercise 3 Bacteria in the Environment

Experiment A: Investigating Specific Environments
In a team of 4, select one of the “environments” below. Use a slightly dampened sterile
cotton swab to collect a sample. Then perform the streak technique to isolate colonies as
shown in your manual.
Hypothesize about the presence of bacteria and fungi in different environments (1pt)

Predict the results of the experiment based on your hypothesis (if/then) (1pt)

Observe the prepared agar plates. Record observations below (6 pts)

Environment Colony Type(s); Abundance

Shoe

Countertop

Stairway Handrail

Rim of Trashcan

Chair Seat

Elevator Button
Exercise 4 Controlling the Growth of Bacteria
Lab Study A. Using Antibiotics to Control Bacterial Growth
In your group of 4, select one of the following bacteria:
Escherichia coli
Serratia marcescens
Staphylococcus aureus
Bacillus subtilis

Using a sterile swab, apply a bacterial lawn on an agar plate. Apply bacteria evenly over
the entire surface. Rotate the plate and swab the plate. Note: the more evenly applied the
bacteria, the better the results.
Select 4 of the antibiotic discs provided (and use one control) to place on the plate as
shown.

Observe the prepared agar plates.

Measure your zone of inhibition (diameter) in mm. Compare to class data. Use the
following criteria to rank bacterial sensitivity of antibiotics:
NS= Not sensitive, no zone of inhibition;
S= sensitive, zone size 0-10 mm;
VS= very sensitive, zone size >10 mm. (16 pts)

Bacteria Antibiotic
Erythromycin Penicillin Tetracycline Control
Escherichia
coli

Serratia
marcescens
 Staphylococcu
s aureus

Bacillus
subtilis

Were the results different for gram-positive and gram-negative bacteria? Describe any
discrepancies. (2 pts)

Lab Study B. Using Antiseptics and Disinfectants to Control Bacterial Growth

In your group of 4, select one of the following bacteria:
Escherichia coli
Serratia marcescens
Staphylococcus aureus
Bacillus subtilis

Using a sterile swab, apply a bacterial lawn on an agar plate. Apply bacteria evenly over
the entire surface. Rotate the plate and swab the plate. Note: the more evenly applied the
bacteria, the better the results.
Select 4 of the disinfectant discs provided (and use one control) to place on the plate as
shown.

Observe the prepared agar plates.

Measure your zone of inhibition (diameter) in mm. Compare to class data. Use the
following criteria to rank bacterial sensitivity of antibiotics:
NS= Not sensitive, no zone of inhibition;
S= sensitive, zone size 0-10 mm;
VS= very sensitive, zone size >10 mm. (16 pts)

Bacteria Antiseptic/Disinfectant/Control
Cavicide Hydrogen Windex Control
Peroxide
Escherichia
coli

Serratia
marcescens

Staphylococcu
s aureus

Bacillus
subtilis

Define antiseptic. (1 pt)

Define disinfectant. (1 pt)