SECTION: BSN -1H DATE: MARCH 11, 2021

LAST NAME, FIRST NAME MI: GUIMALAN, FLOREMER F.

ACTIVITY 6 

ISOLATION OF BACTERIA IN PURE CULTURE 

(Streak Plate Method) 

Objectives:  
At the end of the activity, the student shall be able to: 
a. Demonstrate the presence of various bacteria in the environment and normal flora in
humans. 
b. State the growth requirements of various bacteria and the use of special media for growth of
fastidious organisms. 
c. Isolate pure culture of bacteria from the environment and clinical specimens.
d. describes the colonial morphology of bacteria isolated. 
e. distinguishes between mixed culture and pure culture. 
f. cites the different types of culture media and their uses. 
  g. explains the process of isolating bacteria from clinical specimens in pure culture.
Materials: 
a. Sterile cotton swabs e. NSS solution 
b. Trypticase soy agar (TSA) plates f. Wire loops 
c. Alcohol lamp g. Incubator 
d. Specimens (as assigned) h. Gram stain set-up  

Procedure: 

First Meeting: 
1. Sterilize the inoculating loop in the Bunsen burner by clicking on the loop and dragging it to the
burner.  Put the loop into the flame until it is red hot. Allow it to cool.  
2. Pick an isolated colony from the agar plate culture and spread it over the first quadrant (approximately
1/4 of the plate) using close parallel streaks.  
3. Flame the loop.  
4. Turn the plate 90° and lightly sweep the loop 1-2 times through the inoculated area, then streak into the
next quadrant without overlapping the previous streaks.  
5. Flame the loop. 
6. Turn the plate 90°, overlap the previous area 1-2 times, and streak into the next quadrant (step
4).  7. Flame the loop. 
8. Repeat #6, streaking the remainder of the plate.  
9. Invert the plate and incubate at 37°C for 24 hr.
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Illustration of the Four-quadrant Streak Plate Method  

Second Meeting  

a. Examine the plate and describe the different colonies. Identify the predominant colonies. 
b. Perform Gram stain of the predominant colonies. 

Conclusion: 

Questions: 

1. Define fastidious organisms. 

2. Give the different phases of bacterial growth and describe each phase?
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3. Give the types of culture media according to consistency? 

4. Give the purpose and examples of the following media: a.

Enrichment 

b. Supportive 

c. Selective  

d. Differential

activity 7