MEDIA MAKING

By :
Name : Fitria Ramadhani Ayuningtyas
SID : B1B017046
Entourage :I
Group :5
Assistant : Ainiyya Alfiani

PRACTICAL REPORT OF PHYTOPATOLOGY

MINISTRY OF EDUCATION AND CULTURE

UNIVERSITY OF JENDERAL SOEDIRMAN
FACULTY OF BIOLOGY
PURWOKERTO
2020
 I. INTRODUCTION

A. Background

Microorganisms are found everywhere, almost everywhere. Some

microorganisms are harmful and some are not. Microorganisms can reproduce
naturally or with human assistance. Microorganisms developed by humans
include media. The medium used to grow and reproduce these microorganisms
must be in accordance with the needs of these microorganisms, therefore it is
necessary to know the types of media, how to make the media, as well as
knowing the materials and composition used and the function of each material in
helping the growth of these microorganisms [CITATION Pra08 \l 1033 ].
The survival and growth of microorganisms is influenced by the presence
of nutrients and environmental factors. Nutrients taken from the environment are
then transformed through the plasma membrane to the cell. The nutrients in the
cells are processed to produce energy which is used in cellular processes. The
available nutritional ingredients can be in the form of natural ingredients and it
can also be synthetic materials. The nutritional ingredients used by
microorganisms are usually simple compounds that are available directly or
come from complex compoundswhich are then broken down by microorganisms
into simple compounds through enzymatic processes. These nutrients can be in
the form of liquids or semi-solid solids (semi-solid) which is referred to as
media [ CITATION Lim98 \l 1033 ].
Microorganism growth media is a material consisting of a mixture of food
substances or nutrients needed by microorganisms for their growth. The medium
functions for the growth of microorganisms by isolating microorganisms into
pure cultures and also manipulating the composition of the growth media. Water
(H2O) is the basic material as solvent from agar (seaweed) where the agar
functions as a medium compactor. Formulating a medium or material that will
be used to grow microorganisms in it must pay attention to various kinds
components needed by these microorganisms [ CITATION Sch93 \l 1033 ].
B. Objectives

The objective of this practical laboratory is to find out how to make culture
media for microorganisms.
 II. METHODS

A. Tools
The tools used in the media making lab are heating stoves, autoclaves,
heating pans, erlenmeyer, LAF (Laminar Air Flow), hotplates and magnetic
stirrers.
B. Materials
Materials used in the media-making lab were 200 grams of potatoes,
1700 mL of distilled water, 20 grams of dextrose, 15 grams of agar, cotton, and
wrapper.
C. Procedure

200 grams of potato cut into cubes measuring 1x1 cm. Potatoes are boiled in 500
ml of distilled water until soft.

Potato water is taken by filtering and then put into a 1000ml erlenmeyer then put
in 15 grams of dextrose and 15 grams of agar powder / bars, add distilled water
until the volume becomes 1000 ml and then homogenized using a magnetic
stirer and heated using a hot plate.

The media is put into an Erlenmeyer flask, covered with sterile cotton and
covered again using aluminum foil.
 Media was sterilized using an autoclave at a temperature of 121 ºC with a
pressure of 2 atm for 15 minutes.

The media is removed and allowed to cool slightly, then poured into a petri dish,
or test tube aseptically in the LAF to avoid contamination. Let the media sit in
LAF until it is solid then wrap it. After that, put it in the storage box and ready to
use.

III. RESULT AND DISCUSSION

A. Result

Figure 3.1 Potato Dextrose Agar Instant Figure 3.2 Potato

Figure 3.3 PDA Media

B. Discussion
 Microorganisms can be grown and developed on a substrate called the
medium. Medium is a material consisting of a mixture of nutrients (food
substances) that are used to grow microorganisms. The medium used to grow
and reproduce these microorganisms must be in accordance with the
requirements of the types of microorganisms concerned. The medium can also
be used for isolation, reproduction, testing of physiological properties and
counting the number of microbes. Microorganisms make use of the nutrients in
the medium in the form of small molecules to compose cell components. The
media serves as a place to live, a source of food, and a provider of nutrition for
microorganisms to be cultured on the media [ CITATION Pra081 \l 1033 ].
Based on its physical properties, the medium is divided into three, namely
solid medium, semi-solid medium, and liquid medium. Solid medium is media
containing 15% agar so that after cold the medium becomes solid. Semi-solid
medium is a medium containing 0.3-0.4% agar so that it becomes slightly
chewy, not solid, not so liquid. Semi-solid medium is made so that microbial
growth can spread throughout the medium but does not experience perfect
mixing if shaken, while the liquid medium is a medium that does not contain
agar. The medium based on its shape is divided into three, namely upright test
tubes, oblique test tubes, and petri dishes. The test tube medium is upright,
namely a test tube filled with a medium volume of approximately 6-8 ml. The
test tube medium is oblique, that is, the test tube is filled with a volume of
approximately 3-5 ml. The medium in the form of a petri dish is a petri dish
filled with a volume of approximately 8-10 ml. The medium based on its
composition consists of Synthesis medium, semi-synthesis medium, and non-
synthetic medium. Synthesis medium is a medium whose chemical composition
is known for its exact type and dose. Semi-synthetic medium is a medium whose
composition is partially known with certainty, while non-synthetic medium is a
medium made with an unknown composition [ CITATION Had93 \l 1033 ].
Medium is the material used to grow microorganisms on or in it. Apart from
providing suitable nutrients for microorganisms, physical conditions that allow
for optimum growth are also required. Microorganisms not only vary widely in
their nutritional requirements, but also show different responses to the physical
conditions in their environment. In order for microorganisms to grow well in a
medium, the medium must meet the requirements, among other things, it must
contain all nutrients that are easy to use by microbes, must contain substances
that can inhibit microbial growth, must be in a sterile state before being used so
that microbes what you want to grow well, must have an osmotic pressure,
surface tension and pH that is in accordance with the needs of the microbes
being grown [ CITATION Pel86 \l 1033 ].
PDA (Potato Dextrose Agar) is a medium that is generally used for fungal
growth in laboratories because it has a low pH (pH 4.5 to 5.6), thus inhibiting
bacterial growth which requires a neutral environment with a pH of 7.0 and an
optimum temperature for growth between 25-30 ° C. PDA media including semi
synthetic media because it is composed of natural and synthetic materials. PDA
media are generally known for their chemical composition and are of high
purity. The composition of PDA media is potato, dextrose, and agar. Potatoes
have a function as a source of carbon (carbohydrates), vitamins and energy.
Dextrose functions as a source of sugar and energy, while agar serves to
compact the PDA medium. Each of the three components is necessary for the
growth and reproduction of microorganisms, especially fungi [ CITATION Oct17 \l
1033 ].
Making PDA medium is by peeling potatoes and weighing 200 grams, then
washing them clean, and cutting them into pieces approximately 2 x 2 x 2 cm in
size. The potatoes are boiled with 500 ml of distilled water in a 1,000 ml beaker
glass until the potatoes are soft. The boiled water for potatoes is filtered by
means of a filter and the results of the filter are accommodated in a 1000 ml
volume beaker glass. Put into a dextrose tube as much as 20 gr, 16 g agar and
distilled water until the final volume is 1,000 ml, then heated using a hot plate
and homogenized using a stirrer. The medium after being homogeneous was
then poured into 10 ml test tubes for each tube then covered with cotton and
sterilized (Maharani et al., 2014).

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