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Isolasi Mikroba

The document discusses methods for inoculating microbes onto growth media including streak plating, pour plating, and spread plating. These techniques aim to isolate pure cultures of microbes from samples by diluting and separating individual cells to grow into distinct colonies.

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hizbi01
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51 views

Isolasi Mikroba

The document discusses methods for inoculating microbes onto growth media including streak plating, pour plating, and spread plating. These techniques aim to isolate pure cultures of microbes from samples by diluting and separating individual cells to grow into distinct colonies.

Uploaded by

hizbi01
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Inoculation

 is an attempt to grow microbes from one source


(Sample) into sterile growth media.

 Is introducing microorganisms into a medium where


they can grow and reproduce

 Culture. Microbes that grow together with the


medium.

 Pure culture : if the microbe that grows is


1 type
 Mix culture : if microbes > 1 type

 Isolat is a pure culture of microbes that come from


one type
 isolation is an attempt to get isolate.
 Isolation source → Inoculation → Mix
Culture → Isolation → Isolate (pure
culture)

 Mix culture Pure culture


CULTURING/ INOCULATION

On artificial media

The purpose of growing, breeding, storing


microbes

Requirements for microbial growth :

suitable medium
pH
temperature
humidity
Oksigen + or no
METHODE OF BACTERIA INOCULATION ON
ARTIFICIAL MEDIUM

1. Do aseptically. Avoid the contamination

all materials and tools used must be


sterile

medium must steril : otoklaf, filter,tyndalisasi

work is done near the fire


Ideally : in a Laminar flow
2. Materials and tools prepared for the inoculating
process Material:
Spiritus brander, loop (oese), micropippete (a) .
Sterile medium : solid (b), liquid(c), semi solid (d),
culture / sample
a b c d
 1. Materials taken as samples
 A. Specimens
› B. Culture

 2. Inoculating material on the media


 3. Incubation
saliva Serum/blood Skin Urin

Crevikular plaque jar karies pus periodontal tissue

Dan sebagainya
Swab
.

Methode
of
sampling
speciment
Swab

Scraped
Aspiration
off
B

Incubator

Streaked Plate Technique


Poured plate Technique
Spreader Technique
Microorganisms should be cultured under conditions
that best support their growth including atmosphere,
temperature, and duration of incubation

 An incubator is a device used to grow and


maintain microbiological culturesThe incubator
maintains optimal temperature, humidity and other
conditions such as the CO2 and oxygen content of the
atmosphere inside

 Temperatur is setting in 37o C, 24,48 hour etc

 For anaerobic microba : put in anaerobic jar

anaerbic jar
The purpose is
to dilute out and separate microbe present to get isolated
coloni from a large number of a different microbes

Separated individual cells from large numbers of microbes


and to allow these cells to grow into pure cultures

Technique of Microbial Isolation :

A. Streaked Plate Technique


B. Poured plate Technique
C. Spread plate Technique
Steps of Microbial Isolation

Mucosa of the tongue 2. Put these


sampel in 3.Inoculated
(swab) liquid media in agar plate
medium

Streak /
 Skin : swab Pour plate/
Spreading
Water

Air INCUBATION
Open the agar plate

Separate individual cell from largr number of microbe


ISOLATE ONE TYPE OF MICROBES

PURE CULTURE
Inoculated in agar plate medium

1. STREAK METHODE
Streak the loop in a zig zag patern on the agar plate

take the inoculum


using loop(oese)

There are 4 types of methode used to streak a


plate:

1. T Streak 2. Radiant Streak

Streak the inoculum on


the agar media
With certain pattern

3. Quadrant Streak 4. Continuous Streak


Inoculated in agar plate medium

The way of Sreaking


T streak Continuous streak

 Radiant Streak Quadrant Streak


Inoculated in agar plate medium

 Appearance of coloni after incubation


 T - Streak

Continuous streak Quadrant Streak


Inoculated in agar plate medium

2. POUR PLATE METHODE


The bacterial culture and liquid agar medium are mixed
together.

After mixing , the medium containing the culture poured into


sterilized petridishes , allowed solidifying and then incubated.

After incubation colonies appear on the surface.


PROCEDURE OF POUR PLATE

Melt agar in waterbath, get a melted agar,


Transfer a singel loop of the inoculum to the melted agar

2
Inoculated in agar plate medium

3. SPREAD PLATE METHODE


In this technique, 0,1 ml of sample taken from diluted mixture of culture ,
which is placed on the surface of the agar plate and spread evenly
over the surface by using L spread glass rod called spreader.

Incubate the plates.

After incubation, colonies are observed on the agar place


Appearance of the colonies of air sample after incubation

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