DATA AND OBSERVATIONS:

Skin swabs on Blood Agar.

Observation: 8 yellow colonies forms on the Blood Agar.

Skin swabs on MacConkey Agar

Observation: No colony formed.
 Pus sample on Blood Agar
Observation: Two types of colony formed on the Blood Agar, yellow colonies and colourless
colonies.

Pus Sample on Mannitol Salt Agar.

Observation: Colonies formed surround the agar.
 S. aureus S. pyogenes

Hemolysis on
Blood Agar

S. aureus: yellow colonies S. pyogenes: colourless colonies

Catalase

No bubbles formed. No bubbles formed.

QUESTIONS:
1. The blood agar had to be sterile. How do you sterilize blood?
In order to sterilize the blood for Blood Agar, sterile defibrinated blood that has been
warmed to room temperature was added to the Blood Agar Base. Then the flask was
swirled to mix thoroughly avoiding the formation of bubbles. The suspension was
dispensed into sterile plates carefully avoiding the formation of bubbles.

2. For catalase test, it is not recommended to use cultures from a blood agar plate.
Why?
Because red blood cells contain catalase, will give false-positive result to the catalase
test.

3. In the catalase test, a slow reaction with a few bubbles is not considered positive.
Why?
Because some bacteria have other enzymes that also can decompose peroxide, so few
tiny bubbles forming after 20-30 seconds is not considered positive.

4. In this practical, what are the functions of the blood agar, MacConkey agar and
MSA?
Blood Agar: to detect the hemolysis of skin microflora, to isolate between S. aureus and
S. pyogenes in pus sample and to differentiate between colonies of S. aureus and
colonies of S. pyogenes on Blood Agar.

MacConkey Agar: to isolate gram-negative bacteria on skin swab.

MSA: to isolate pathogenic staphylococci on pus sample.

5. You received a pus sample and was asked to check if it contains S. aureus. How
will you do it to obtain results in shortest time? Draw your flowchart on the blank
page opposites.
REFERENCES

1. Blood Agar Plates and Hemolysis Protocols. (2005). Retrieved from

https://www.asm.org/getattachment/7ec0de2b-bb16-4f6e-ba07-2aea25a43e76/protocol-
2885.pdf.

2. McFaddin, J. F. (2000). Biochemical tests for identification of medical bacteria.

Philadelphia: Lippincott Williams & Wilkins. Retrieved from
http://staff.um.edu.mt/amce1/Stains and Tests PAT2322/Catalase test.pdf.

3.