Lesson Plan: Bacteria & Viruses

Name: Ms. Desmond Date: ~3 days

Subject: Intensive Science Grade Level: 9
Class Period: 71 minutes Materials: Who Dun It Lab, Bacteria & Viruses
Lecture, Bacteria & Viruses Guided Lecture Notes,
Gram Stain Lab

Standards:
B2.4h Describe the structures of viruses and bacteria.
B2.4i Recognize that while viruses lack cellular structure, they have the genetic material to invade living cells.
Objectives:
Students can model how bacteria is passed through body fluids
Students can compare and contrast bacteria and viruses
Students can recall the lytic and lysogenic cycle of viruses
Students can differentiate between gram + and gram – bacteria

Introduction/Hook: Who Dun It: A Demonstration (see handout for details)

Steps in the Lesson:
Day 1: After the “Who Dun It” Lab, students will receive the “Bacteria & Viruses Guided Lecture Notes”
worksheet. The students will follow along while the lecture is given. At the end of this class period, part I of
the “Gram Stain Lab” needs to be completed.
Day 2: The lecture will be finished, and the process of gram staining bacteria will be explained in further
detail to prepare the students for the next day’s lab.
Day 3: Students will need the full 71 minutes to complete the “Gram Stain Lab”
 Bacteria and Viruses Guided Lecture Notes
Describe the characteristics of each Domain:
1. Archaea

2. Bacteria

3. Eukarya

What are extremophiles? Give some examples:

How are bacteria different from Archaea?

Cell shapes
1. Cocci

2. Bacilli

3. Spirilli

Describe Cell wall Structure.

What does it mean when a bacteria gram stains PURPLE?

What does it mean when a bacteria gram stains PINK?

How do bacteria move?

Bacterial Groupings: Describe and DRAW

1. Singles:

2. Doubles:
 3. Clusters:

4. Lines:

What is binary fission? Draw it.

What is conjugation?

Bacterial Nutrition:
1. Photoautotroph

2. Chemoautotroph

3. Photohetertroph

4. Chemoheterotroph

List some benefits of bacteria.

List some concerns about bacteria

Viruses: Describe their structure.

How are viruses classified?

Describe the lytic cycle:

Describe the lysogenic cycle.

List some viral diseases:

 GRAM STAIN LAB
NAME: _____________________________
INTRODUCTION:
Most common bacteria are either Gram-positive or Gram-negative (based on cell wall
structure). Remember from lecture, Gram-positive cell walls consist of several layers of
peptidoglycan. Gram-negative cell walls have one layer of peptidoglycan surrounded by a lipid-
based outer membrane. In
the 1880s, Hans Gram developed the differential method of staining that bears his name. While
we still don’t know exactly why Gram-positive bacteria end up looking different from Gram-
negative bacteria, Gram-staining is still an important way to characterize bacteria.

Gram-staining begins by getting cells to stick on a clean microscope slide. Such a prep is called
a bacterial smear. To a bacterial smear the following chemicals are applied to make a Gram-
stain:

PURPOSE: To determine the basic forms bacteria can take and the Gram quality of the cell
membrane.

MATERIALS:
agar broth clothes pin
petri dishes crystal violet stain
distilled water Gram’s iodine
cotton applicator ethyl alcohol – 95%
inoculating loop safranin stain

Part I: Collection of Bacteria for Gram Stain

1. Wet tip of cotton applicator in distilled water.

2. Rub across surface to be tested.
3. Carefully tilt lid of petri dish open and rub cotton applicator across surface of agar.
4. Tape lid closed.
5. Label with name, hour, and location of collection.
6. Place in incubator for 3 days at 33° C.

Part II: Culture Transfer

1. Wash, rinse and dry a slide.

2. Pass the end of the inoculating loop through the flame of a Bunsen burner until it is
red hot.
3. Let it cool.
4. Put a drop of water on the slide.
 5. Use the inoculating loop to pick up a small amount of the bacteria culture from the
petri dish.
6. Add the bacteria on the loop into the water drop on the slide, mixing it thoroughly,
and spreading it out into a large, thin layer.
7. Air dry the slide on the warming tray until the water is gone.
8. Holding the slide in a clothespin, heat fix the bacteria by passing the bottom of the
slide through the flame of the Bunsen burner for 5 seconds.
9. Let it cool. Leave the clothespin on it for the staining portion.

Part III: Staining

1. Gram’s Crystal Violet Stain - Holding slide with clothes pin over sink, flood smear with
Crystal Violet for 60 seconds. Crystal violet is a purple chemical that sticks to the
peptidoglycan layer of the bacterial cell wall. After 60 seconds, crystal violet is rinsed off using
tap water.

2. Iodine- Flood slide with Iodine for 60 seconds. The iodine is called a mordant—it causes
crystal violet to stick to peptidoglycan like mortar causes bricks to stick together. After 60
seconds, iodine is rinsed off using tap water.

3. Acetone/Ethanol Wash – Flood slide with Acetone/Alcohol for JUST 5 SECONDS. The
Acetone/Alcohol washes crystal violet out of the Gram-negative cell wall. We’re not really sure
why this happens, but it does. The Gram-positive cell wall retains crystal violet as long as the
acetone/alcohol wash lasts not more than a few seconds. After 5 SECONDS, the
acetone/alcohol is rinsed off using tap water. The acetone/alcohol wash is the differential step in
the Gram-stain process. That is, it is the acetone/alcohol that creates the observable difference:
Gram-positive cells look purple after this step; Gram-negative cells look clear.

4. Safranin Stain - Flood slide with Safranin for 60 - 90 seconds. Safranin is a pink stain that
sticks to cytoplasmic components of the cell. All cells become stained with Safranin. Gram-
positive cells are pink on the inside, but you can’t see this because they are dark purple on the
outside (kind of like a bon-bon). Gram negative cells, which were cleared in the previous step,
end up looking pink. After 90 seconds to two minutes (the longer the better), Safranin is rinsed
off with tap water.

5. Allow slide to dry. Once dry, the slide is ready for observation.

Part IV – Observing Slide

1. Place slide under microscope at lowest magnification.
2. Record for each slide:
Type of bacteria - __________________
Gram positive or negative____________
Grouping - _____________
 Who Dun It: a Demonstration
Prep:
• From the lab get: disposable pipets, phenolphthalein, sodium hydroxide and Dixie cups.
• Select one Dixie cup and fill ¾ full of sodium hydroxide.
• Prepare enough Dixie cups ¾ filled with water so each class member will have one.
• Give each child a pipet.
• Pass out cups to each class member (remember who receives the sodium hydroxide). If
the class size is odd, you will have to participate.

Rules:
Move into an open area like the rotunda. Explain to the kids that someone in the class has
contracted a very serious deadly disease (your choice). It is passed to new people by the
exchange of body fluids. Then have them find a partner. Fill their pipet from their cup and
squirt it into the other person’s cup. Then have them wander around the room. When you say
stop, then need to find a new partner. Exchange body fluids. Repeat two more times so they
have exchanged a total of four times. Return to class. As they enter into class, put a drop of
phenolphthalein into each cup. If they turn pink they died; if clear, they survived.

Results:
Record on the board each person who died (turned pink).
Record the people (in order) that each dead person exchanged with.
Use logic to work backwards to determine who was the initial sick person. You should narrow
it down to two. Then you will have to tell them who it really was.

Disposal:
Throw all pipets and cup away.

This is a fun way to look at the spread of disease and at how the CDC actually determines who
it was that was patient 1.