Basra University College of science and

Technology
Pharmacy department

Medical Microbiology

Title of Report:

streak plate cultivation

Students Name:
‫مريم يونس عبد الجبار صالح‬

Supervised by: Dr: Zainab safaa

the theory:

In microbiology, streaking is a technique used to isolate a pure strain

from a single species of microorganism, often bacteria.

He introduced solid media for culture of bacteria. Koch pioneered the use
of agar as a base for culture media. He developed the pour plate method
and was the first to use solid culture media for culture of bacteria.

Koch also developed media suitable for growing bacteria isolated from
the body. Because of their similarity to body fluids, meat extracts and
protein digests were used as nutrient sources. The result was the
development of nutrient broth and nutrient agar media that are still in
wide use today.

He also introduced methods for isolation of bacteria in pure culture.

He described hanging drop method for testing motility.

He introduced staining techniques by using aniline dye.

He invented the hot air oven and steam sterilizer, and also introduced
methods to find out the efficacy of antiseptics.

The dilution or isolation by streaking method was first developed by

Loeffler and Gaffky in laboratory, which involves the dilution of bacteria
by systematically streaking them over the exterior of the agar in a Petri
dish to obtain isolated colonies which will then grow into the number of
cells or isolated colonies.

Streaking is rapid and ideally a simple process of isolation dilution.

The technique is done by diluting a comparatively large concentration of
bacteria to a smaller concentration.

The decrease of bacteria should show that colonies are sufficiently

spread apart to affect the separation of the different types of microbes.

Streaking is done using a sterile tool, such as a cotton swab or

commonly an inoculation loop.

Aseptic techniques are used to maintain microbiological cultures and to

prevent contamination of the growth medium.

The streak plate method is a rapid qualitative isolation method.

The techniques commonly used for isolation of discrete colonies

initially require that the number of organisms in the inoculums be
reduced.

It is essentially a dilution technique that involves spreading a loopful of

culture over the surface of an agar plate.

The resulting diminution of the population size ensures that, following

inoculation, individual cells will be sufficiently far apart on the surface of
the agar medium to affect a separation of the different species present.

In the streaking procedure, a sterile loop or swab is used to obtain an

uncontaminated microbial culture. The process is called “picking
colonies” when it is done from an agar plate with isolated colonies and is
transferred to a new agar or gelatin plate using a sterile loop or needle.

The inoculating loop or needle is then streaked over an agar surface.

On the initial region of the streak, many microorganisms are deposited

resulting in confluent growth or the growth of culture over the entire
surface of the streaked area.
The loop is sterilized by heating the loop in the blue flame of the Bunsen
burner, between streaking different sections, or zones and thus lesser
microorganisms are deposited as the streaking progresses.

The streaking process will dilute out the sample that was placed in the
initial region of the agar surface.

Methods of Streak Plat:

There are many different types of methods used to streak a plate. There
are two most commonly used streak patterns, a three sector “T streak”
and four-quadrant streak methods.

Picking a technique is a matter of individual preference and can also

depend on how large the number of microbes the sample contains.

T-Streak:
The three-phase streaking pattern is known as the T-Streak.

The streaking is done using a sterile tool, such as a cotton swab or

commonly an inoculation loop.

The inoculation loop is first sterilized by passing it through a flame.

When the loop is cool, it is dipped into an inoculum such as a broth or

patient specimen containing many species of bacteria.

The inoculation loop is then dragged across the surface of the agar back
and forth in a zigzag motion until approximately 30% of the plate has
been covered.

The loop then is re-sterilized and the plate is turned 90 degrees.

Starting in the previously streaked section, the loop is dragged through it
two to three times continuing the zigzag pattern.

The procedure is then repeated once more being cautious to not touch
the previously streaked sectors.

Each time the loop gathers fewer and fewer bacteria until it gathers just
single bacterial cells that can grow into a colony. The plate should show
the heaviest growth in the first section.

The second section will have less growth and a few isolated colonies,
while the final section will have the least amount of growth and many
isolated colonies.

Quadrant method:
In the quadrant method, four equally sized sections are streaked. The
continuous streaking method typically involves inoculating the top half of
the plate, rotating it 180 degrees, and inoculating the other half of the
plate without sterilizing the loop or dragging bacteria from the previous
section.

Discontinuous Streaking Method:

1-Sterilize the inoculating loop in the bunsen burner by putting 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 or insert your loop into the tube/culture bottle and remove some
inoculum. You don’t need a huge chunk.
3-Immediately streak the inoculating loop very gently over a quarter of
the plate using a back and forth motion.

4-Flame the loop again and allow it to cool. Going back to the edge of
area 1 that you just streaked, extend the streaks into the second quarter of
the plate.

5-Flame the loop again and allow it to cool. Going back to the area that
you just streaked, extend the streaks into the third quarter of the plate.

6-Flame the loop again and allow it to cool. Going back to the area that
you just streaked, extend the streaks into the center fourth of the plate

7-Flame your loop once more

Significance of Streak Plate Method:

The streak plate technique is the most popular method for isolating
specific bacteria from a sample containing a mixture of microorganisms.

Streak plate technique is used to grow bacteria on a growth media

surface so that individual bacterial colonies are isolated and sampled.

Samples can then be taken from the resulting isolated colonies and a
microbiological culture can be grown on a new plate so that the organism
can be identified, studied, or tested. When the bacteria are streaked and
isolated, the causative agent of a bacterial disease can be identified.
Limitations of Streak Plate:
Streak plating is a microbiology laboratory method that has two major
disadvantages.

Firstly, users will not be able to grow obligate anaerobes using this
method.

Secondly, only organisms that were viable in the original sample are
able to be grown.

Results:
The streaked plate is incubated at 37°C for 24 hours. Examine the
colonies grown in the plate carefully. All colonies should have the same
general appearance. If there is more than one type of colony, each type
should be streaked again on a separate plate to obtain a pure culture.