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3 views

isolation

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bvmg8vh9xd
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Isolation of bacteria in pure culture

A. I. Mustafa Adnan Al. Noori


 Isolation: To separate a single
species of bacteria from a mixed
population.
 Pure culture consists of only a single
type of organism.
Aim:
To identify the bacterial pathogens
Culture method
 Usage of selective and differential media
1.Streak plate technique
• Isolation of bacteria in pure culture from clinical
specimen
2.Spreading‐plate method:
*Quantification of bacteria in liquid cultures, urine
sample.
*Swabbing from clinical samples
3.Pouring‐plate method:
 This is the most commonly used method for
enumeration of bacteria in a wide variety of
samples including milk, food, meat, soil etc.

 pour plate methods yield a count of only the living


cells in the sample and thus are a viable count.
Culture method

4.Stabbing method:
• To differentiate between aerobic
and anaerobic bacteria.
• differentiate between motile and
non‐motile bacteria
Streak Plate Isolation Principle
• An original inoculum containing a mixture
of bacteria is spread into 4 quadrants on
solid media.
• The goal is to reduce the number of
bacteria in each subsequent quadrant.
Streak Plate Isolation Principle
• Colonies are masses of offspring from an
individual cell therefore streaking attempts
to separate individual cells.
• Discrete colonies form as the individual
cells are separated and then multiply to
form isolated colonies in the later
quadrants.
Spread Plate Method
• Spread plate method- small volume of liquid,
diluted sample pipette on to surface of the
medium and spread around evenly by a sterile
spreading tool
Pour Plate method:
• There are two steps to the process:
 Dilution of the sample so that various dilutions
of the sample may be inoculated onto plates and
a count of the colonies that grow made;
 The second step is the plating of the dilutions so
that each cell in the diluted sample may then
grow and form colonies that will in turn become
visible to the naked eye and can be counted.
Procedure:

- Aseptically dilute sample with sterile water blanks


- Make 1:10 dilutions as instructed, based on sample
- Aseptically transfer 1ml volume of dilutions that are to
be plated onto sterile Petri plates
- Pour molten agar onto the sample in Petri dish and mix
thoroughly
- Incubate as instructed for growth of colonies
- During next lab period, count colonies on plates
Classification of Bacteria according to O2
requirement
1. Aerobic: Pseudomonas.
2. Anaerobic: Clostridia.
3. Facultative anaerobic: Streptococci,
E. coli.
4. Microaerophilic:Helicobacter pylori.

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