Microbiologist: 3.1.5 Isolation & Gram Staining

3.1.
5 Isolation & Gram Staining

Directions: As you go through Lesson 3.1 in CTLS, be sure to follow along and
complete the document. Be thorough, be complete, and be precise. Keep answer font
red or some color other than black or grey. This lesson should take you
approximately 120 minutes to complete. See directions in CTLS for where and how to
submit your work.
PART A: Isolation
How are bacterial species named?

__ To identify bacteria, lab tests
are conducted that look at specific characteristics of the
organisms. The way in which the organisms grow, their morphology, or shape of their cells,
and their ability to metabolize.
Microbiologist
What they do Read about microbiologist, Angela Jones, and her work
__ Plan and conduct complex research at the Mayo Clinic. In her profile, Angela notes that her
projects, such as developing new drugs to greatest challenge is dealing with antibiotic resistant
combat infectious diseases.
bacteria in patients at the hospital. Conduct some
research, if necessary, and record how bacteria develop
a resistance to antibiotics.
__ Antibiotic resistance occurs when bacteria develop the
ability to survive exposure to antibiotics that were designed to
kill them or stop their growth.
Isolation Notes:
__ Antibiotic resistance occurs when bacteria develop the ability to survive
exposure to antibiotics that were designed to kill them or stop their growth.
Agar provides all the nutrients and water bacteria need to grow. The process by which
bacteria are streaked onto a plate to form isolated colonies is called an isolation streak.
Reflection Question: Why is it important to use aseptic technique when working with bacteria in the lab?
__to make sure you’re samples of bacteria don’t get contaminated
If using a metal re-usable inoculating loop, what is the purpose of flaming it and letting the loop cool?
__ to kill any foreign bacteria that may contaminate your sample.
Describe the process of producing an isolation streak on a petri dish.

__ You will streak each patient sample onto an agar plate to physically separate the different
species of bacteria.
Reflection Question: If isolated colonies do not grow on a plate, what may have happened?
__they may have been contaminated by forging bacteria or fungi.
For each patient sample, record the following:

a. How many different species of bacteria appear to be in each patient sample? How do you know?
__in each patient there was a circular form cell and in patient 1 and 2 there was a convex and
in 3 there was an entire.
b. Complete the table:

Colony Characteristics
Patient: Color Form Elevation Margin Size (mm)
1 yellow circular convex none large
2 red circular convex none small
3 yellow circular none entire large
a. Is there one organism that is common between the patients? Justify your answer.
__Yes, circular form because each sample has circle like cells.
Based on your observations, what are the possible identities of the organism causing the hospital outbreak?
How do you know?
__ Bacteria, Because the shapes of the colonies match the ones most commonly found in bacteria.
Reflection Question: Based on this experiment, has your team definitively identified the organism causing
the infection in these patients? If not, what else do you need to know? Is there other testing that needs to be
done?
__ no, there still needs to be gram staining testing for more proof.
Part A Conclusion Questions:
1. When identifying the agent responsible for causing a disease, why is evaluation of colony morphology
not enough?
__there are other forms of bacteria/fungi with similar colony morphology.
2. Why is it so important to determine the genus and species of an organism causing disease? What
could happen if the wrong organism is identified as the cause of disease?
__so you can identify how to deal with it, you could waste time developing cures that don’t
even work.
PART B: Gram Staining
Complete the table below by listing each bacterial cell morphology and draw or SNIP and paste what they
look like.
Cell Type DRAW what what they look like

Coccus
Bacillus
Spirillum
In the space below, list the different bacterial cell arrangements and DRAW what the arrangements look like.
__
Draw and label the structures of Gram positive and Gram negative cell walls. Use a purple colored pencil to
draw the Gram positive peptidoglycan layer and use a pink or red pencil to draw the Gram negative
peptidoglycan layer.
__
What additional feature makes Gram negative more dangerous usually?
__ Gram negative bacteria are often more dangerous because they have an additional layer
outside of their peptidoglycan layer.
Record your observations of the prepared slides (use colored pencils in your drawings):
Slide # Cell Cell Gram Drawing
Morphology Arrangement Results
(cocci, bacilli,
spirilla)
1 Cocci All the cocci positive
forms
2 bacilli Chain of bacilli negative

3 spirilla individual negative
Diagram what happens in each step of the Gram stain.
Record your observations of your patient slide.

Slide Cell Cell Gram SNIP of Image
Morphology Arrangement Results
cocci All forms negative
Patient
a. Record the identity of the organism causing the outbreak.
__ Staphylococcus aureus
b. Conduct research into this organism and record the type of infections typically experienced by those
afflicted by it and its mode of transmission.
i. How do the infections you read about compare to those the 11 affected hospital patients
experienced?
__ Staph infections are caused by staphylococcus bacteria, types of germs commonly found on the skin or in
the nose of even healthy individuals. Most of the time, these bacteria cause no problems or result in relatively minor
skin infections. But staph infections can turn deadly if the bacteria invade deeper into your body, entering your
bloodstream, joints, bones, lungs or heart. The hospital patients are experiencing infected lungs and skin from
bacteria. Mode of transmission is direct and indirect contact
ii. Does the mode of transmission you read about fit the hypothesis your team developed with
regard to how this agent is spreading?
__yes
c. Revisit the treatment recommendation you previously made for the patients. Do you need to update
your recommendation? If so, how?
__no, my recommendations were to wash your hands and isolate yourself and the things you
touch, so its pretty accurate.
d. Compose a message to the staff at GNMH. You need to inform them of your conclusions: the cause
of the infections, the recommended treatment, and the steps to prevent further spread. Keep your
communication short and tactful.
__Dear hospital, we have Identified the disease and we are sending treatment plans and
medication over as you read this email.
Conclusion Questions:
1. List general measures GNMH could take to reduce the chance of nosocomial infections of any type.
Reflect on all the information you have learned in previous activities as well as the information you
have accessed through the Hospital Hub app.
__Refer to the previous email made when we first contacted you.
2. If you and your teammates saw different Gram stain results, what might this mean? Why might this
have occurred? (Even if it didn’t happen to you – what would it have meant?)
__we might have listed a different disease and had to made different trament plans and
procedures.

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What are the steps to properly isolate bacterial colonies?

Why is it important to identify the specific bacteria causing an infection?

3.1.

5 Isolation & Gram Staining

How are bacterial species named?

Describe the process of producing an isolation streak on a petri dish.

For each patient sample, record the following:

b. Complete the table:

2 red circular convex none small

3 yellow circular none entire large

Part A Conclusion Questions:

PART B: Gram Staining

Cell Type DRAW what what they look like

2 bacilli Chain of bacilli negative

Diagram what happens in each step of the Gram stain.

Record your observations of your patient slide.

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