GEBZE TECHNICAL UNIVERSITY

MICROBIOLOGY LABORATORY REPORT - 2

EBRU AKHARMAN
142204026
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MICROORGANISM CULTURE TREATMENT

TECHNIQUES
AIM:

Microbial cultures are foundational and basic diagnostic methods used extensively as a research tool in
molecular biology. It is often essential to isolate a pure culture of microorganisms. A pure culture is a
population of cells or multicellular organisms growing in the absence of other species or types. A pure
culture may originate from a single cell or single organism, in which case the cells are genetic clones of one
another. For the purpose of gelling the microbial culture, the medium of agarose gel (agar) is used. Agar is
a gelatinous substance derived from seaweed.
Microbiological cultures can be grown in petri dishes of differing sizes that have a thin layer of agar-based
growth medium. Once the growth medium in the petri dish is inoculated with the desired bacteria, the
plates are incubated at the best temperature for the growing of the selected bacteria . Another method of
bacterial culture is liquid culture, in which the desired bacteria are suspended in liquid broth, a nutrient
medium. These are ideal for preparation of an antimicrobial assay.
The various techniques are used in cultivation of bacterias. These techniques are separated to two units.
First units is transfer and colony selection techniques. The subset of first units are from liquid media to a
liquid medium, from solid medium to liquid medium and from solid media to slant agar. Second units is
pure culture isolation techniques. The subset of second units are from solid media to solid media with
streak method, from liquid media to solid media with spreading method and from liquid media to solid
media with casting method. These techniques will be used in this laboratory. The aims of these
experiments are learning different techniques and obtaining pure cultures.
INTRODUCTION:
When microorganisms are cultivated in the laboratory, a growth environment called a medium is used. The
medium may be purely chemical (a chemically defined medium), or it may contain organic materials, or it may
consist of living organisms such as fertilized eggs. Microorganisms growing in or on such a medium form
a culture. A culture is considered a pure culture if only one type of organism is present and a mixed culture if
populations of different organisms are present. When first used, the culture medium should be sterile,
meaning that no form of life is present before inoculation with the microorganism. Pure culture,
in microbiology, a laboratory culture containing a single species of organism. A pure culture is usually derived
from a mixed culture (one containing many species) by transferring a small sample into new, sterile growth
medium in such a manner as to disperse the individual cells across the medium surface or by thinning the
sample manyfold before inoculating the new medium. Both methods separate the individual cells so that,
when they multiply, each will form a discrete colony, which may then be used to inoculate more medium,
with the assurance that only one type of organism will be present. Isolation of a pure culture may
be enhanced by providing a mixed inoculum with a medium favouring the growth of one organism to the
exclusion of others.
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from liquid media
to a liquid medium

Colony Selection from solid medium

Techniques to liquid medium

from solid media to

slant agar
CULTURE
TREATMENT
TECHNIQUES from solid media to
solid media with
streak method

from liquid media

Culture Isolation
to solid media with
Techniques
spreading method

from liquid media

to solid media with
casting method

Colony Selection Techniques

 1.A. From liquid media to a liquid medium :

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 2.A. From solid medium to liquid medium :

 3.A From solid media to slant agar

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 4.A From solid media to solid media with streak method

 5.A From liquid media to solid media with spreading method

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 6.A From liquid media to solid media with casting method

RESULTS
 1.B: Experiment 1

Bacterial cultures are prepared in liquid medium to identify bacteria by any biochemical test and to isolate
DNA, RNA, plasmids. Bacterial cultivation on liquid medium from liquid medium can be done either with
micropipette or with a steril micropipette tip, or with a sterilized micropipette. An intensive breeding is
observed in this tube. When the tube is shaken, the medium is quite cloudy.
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 2.B: Experiment 2

By this culture method, a single colony formed in the solid medium can be selected and the isolation of DNA
from this colony or the transfer of the biochemical characteristics of the colony to the liquid medium can be
carried out. Sterilization from solid medium to liquid medium is accomplished with sterile care. No
proliferation was observed in this tube.

 3.B: Experiment 3

With the slanted agar used, a large surface area is obtained in the tub. Thanks to this feature, this slanting
method is used for many purposes such as bacterial identification, culture collection creation. Slanting of the
solid medium from the medium is accomplished with the help of a sterile needle-tipped nose. No proliferation
was observed in this tube.
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 4.B: Experiment 4

By this culture method, the bacteria are allowed to form individual colonies by streaking on agar in a mixed
bacterial culture. It also learns about the morphology of a single falling bacterial colon. Culture is achieved
with the help of streil special. There is proliferaiton in the petri dish and pure colonies are formed. In addition,
no contamination was observed.

 5.B: Experiment 5

With this method it is possible to obtain a pure culture by finding a single colony and to learn about the
morphology of a single falling colony of bacteria. This culture method is expected to grow bacterium
intensively but no breeding is observed.
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 6.B: Experiment 6

By this method of culture, it is possible to calculate the number of viable cells which can form colonies in
liquids such as water and milk as well as obtain pure culture by lowering single colonies. In addition, hemolytic
activities of some bacteria can be observed with studies performed with bloody agar. No breeding is
observed.

DISCUSSION
Microbial cultures are foundational and basic diagnostic methods used extensively as a research tool in
molecular biology. It is often essential to isolate a pure culture of microorganisms. A pure culture is a
population of cells or multicellular organisms growing in the absence of other species or types. A pure culture
may originate from a single cell or single organism, in which case the cells are genetic clones of one another.
In this experiment various culturing methods were used. As a result of this culture, no reproduction was
observed in many petri dishes and tubes were not observed. The reason for this situation is that the
inoculating loop with fever is not sterile during sterilization or colonies could not be enough or the inoculating
loop is not cooled enough before the colonies is taken. If the inoculating loop is not cooled enough, the
bacteria is died so proliferation is not observed in culture. Another possibility is that the culture is defective. If
the culture is defective, bacteria may not grow.