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

Sterilize the inoculating loop in the bunsen burner by putting the loop -1
.into the flame until it is red hot. Allow it to cool

Pick an isolated colony from the agar plate culture and spread it over -2
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
Immediately streak the inoculating loop very gently over a quarter of -3
.the plate using a back and forth motion

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

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

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

Flame your loop once more-7

: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