Lab 1: Hand Washing to Control Normal Flora

We have it drilled into our heads from the time we are tiny children that we must
wash our hands to get rid of the germs. Yet during the Middle Ages, washing was
considered a method of spreading disease. Even as recently as the turn of the 20th
century, some health care providers considered hand washing to control
contamination nonsense. But does hand washing really remove the organisms on
our hands? Is there a method that works better than others? This experiment is
designed to allow you to make that determination. While you are working here only
with normal flora (organisms that normally live on and in us, usually without
producing disease), the experiment demonstrates the tenacity and contact
transmission of microbes in general.
Materials
petri plate of nutrient agar
marking pen
hot water
soap
clean test tube brush
tape
Procedure
1. Divide the bottom of a petri plate in quarters using the marking pen. Label
the plate with your group name on the edge of the lid. Write small.
2. In small print, label the sections as follows:
o unwashed- your name
o water only- your name
o unwashed- your name
o soap and water- your name
3. Rub the index and middle fingers of one group members hand on the
unwashed quadrant of the petri plate.
4. The group member washes his/her hands under warm running water. Rub
them well, but do not use soap.
5. Shake off the excess water and dry the fingers with a paper towel. Leave
fingers slightly damp.
6. Rub the fingers on the water only quadrant.
7. Another group member rubs his/her unwashed fingers on the agar of the
labeled section.
8. The group member washes his/her hands under warm running water, using
soap
9. Shake off the excess water and dry the fingers with a paper towel. Leave
fingers slightly damp.
10.Rub the fingers over the soap and water quadrant.
11.Tape the petri dish closed by taping around the edges of the pate.
12.Invert the petri plate and place in the incubator.

13.24-72 hrs later, count the number of colonies in each quadrant. Each colony
represents one original bacterial cell.
14. Enter data in a table in your composition notebook.
15.Photograph the plate, and save to the website.
16.Complete the lab requirements as stated on the lab requirement handout.

LAB 2: HOW CLEAN IS YOUR KITCHEN?

With cases of food contamination and food poisoning on the rise, many people ask
how clean their kitchen is. Is E. coli lurking on the counter? Salmonella hiding in the
fridge? Or is your kitchen floor truly clean enough to eat off of? This experiment lets
you examine the microbial populations of your kitchen and identify possible
sources of contamination. You may find the results reassuring or frightening,
depending on what grows on your plates.
Materials

sterile swabs
sterile petri plates
marker
tape
used kitchen towel, used sponge, containers from fridge, silverware from
drawer, egg carton, veggies

Procedure
1. Divide the bottom of a petri plate in quarters using the marking pen. Label the
plate with your group name.
2. Choose 4 items or places to sample from the posted list. Label each quadrant of
the plate with name of the object sampled.
3. To sample your counter, floor, table, phone, sink faucet handle, and cutting
board:
a. moisten a swap with boiled water
b. scrub the swab across the surface to be sampled
c. gently rub the swab across the appropriate side of the agar
4. To sample your sponge, sink drain, and dish towel:
a. use the object as usual
b. after use, touch the damp object to the appropriate side of the agar
5. Other objects in the kitchen you may wish to sample using a damp swab
a. a silverware from drawer
b. back of a chair
c. the handle of the frig
d. an egg shell or the egg carton
e. outside rim of the milk jug / juice container
f. washed and unwashed fruits or veggies
g. T.V. or computer screen or key board
6. Tape the petri dish closed by taping around the edges of the pate.
7. Invert the petri plate and place in the incubator.
8. 24-72 hrs later, count the number of colonies in each quadrant. Each colony
represents one original bacterial cell.

9. Enter data in a table in your composition notebook.

10. Photograph the plate, and save to the website.
11. Complete the lab requirements as stated on the lab requirement handout.

LAB 3: ANTIMICROBIALS
One of the most commonly prescribed drugs are antibiotics. However, different
bacteria have different susceptibilities to antibiotics. The easiest method for
determining susceptibility is a technique called a Kirby-Bauer method. Small disks of
paper with different concentrations and different types of antibiotic are placed on a
lawn of bacteria on an agar plate. The antibiotic then diffuses through the agar.
Susceptibility is determined by measuring the zone of inhibition surrounding each
disk. Basically, the bigger the zone, the more susceptible the microbe is to that
specific antibiotic. In addition, any resistant bacteria will appear as colonies within
the zone of inhibition.
Materials
petri plates of nutrient agar
sterile swabs
forceps
Bunsen burner
striker
discs of cleaners
sterile paper disks
Procedure
1. Divide the bottom of a petri plate in quarters using the marking pen. Label
the plate with your group name.
2. Decide on 3 antiseptics that you would like to test.
Your 4th will be water which will be your control.
3. Label each quadrant of the plate with name of the object sampled.
4. To sample your counter
a. moisten a swap with boiled water
b. scrub the swab across the surface to be sampled
c. gently rub the swab across the appropriate side of the agar
5. Take a paper disk, dip it in the antiseptic and place it
in the middle of the quadrant. See figure 2-31.
6. Tape the petri dish closed by taping around the edges of the pate. Figure 2-31
7. Invert the petri plate and place in the incubator.
8. 24-72 hrs later, measure the zone of inhibition for each quadrant. The zone
of inhibition should be measured in mm. See figure 2-32.
9. Enter data in a table in your composition notebook.
10.Photograph the plate, and save to the website.
11.Complete the lab requirements as stated on the lab requirement handout.

Zone
Zone
of Inhibition
of Inhibition
Figure
Figure
2-32
2-32

Zone of Inhibition
Figure 2-32