Culture Media

1
Contents
Culture media Culture medium, Inoculum, Culture,
a n d some
Cultivation /growth
te rminolo g ie s

Purposes of
Bacterial Some C o m m o n Purposes
cultivation

Ingredients of Water, Agar, Peptone, Meat extract, Yeast extract,

Culture Malt extract, Blood and Serum
medium

C la ssific a tio n On the basis of phases

of culture On the basis of nutritional Factor
media Special m e d i a
2
3 We should know about some terminology…

Culture medium

Inoculum

Culture

Cultivation /growth
 Culture medium:
4 A nutrient material prepared for the growth of
microorganisms in a laboratory is called a
culture medium.
CULTURE : Is the term given to microorganisms that are cultivated in the lab for the purpose of
identifying and studying them. • MEDIUM: Is the term given to the combination of ingredients that
will support the growth and cultivation of microorganisms by providing all the essential nutrients
required for the growth (i.e. multiplication) in order to cultivate these microorganisms in large
number to study them.
5 Inoc ulum:

Whe n microbes are introduced into a culture

medium to initiate growth, they are called an
inoculum.
 Culture:
6 The microb e s tha t gro w a nd multip ly in or on a
culture medium are referred to as a culture.
7 Cultivation

In vivo environment.
In vitro environment.
By appropriate procedures they h a v e to b e grown
separately (isolated) on culture media and obtained as
pure cultures for study.
 PURPOSE OF BACTERIALCULTIVATION
8 Bacterial cultivation has three main purposes:
Grow a n d isolate.

Infection a n d contaminants or colonizers.

Identification a n d characterization .
Reasons for bacterial cultural:
Bacteria have to be grown (cultured) for them to be subsequent clinical diagnosis.
Culturing bacteria is also the initial step in studying its morphology and its
identification.
By appropriate procedures they have to be grown separately (isolated) on culture
media and obtained as pure for study.
Bacteria have to be cultured in order to obtain antigens from developing serological
assay for vaccines.
Certain genetic studies and manipulations of the cells also need that bacteria be
cultured in vitro.
Culturing on solid media is another convenient way of separating bacteria in mixture
9 COMMON INGREDIENTS OF CULTURE MEDIA
Some of the components of culture media are as follows:
Water

Agar

Peptone

Meat Extract

Malt Extract

Yeast Extract

Blood a n d Serum
 1. Water:
10 Tap water is often suitable for culture media.

If the local supply is found unsuitable, glass distilled or

demineralized water must be used instead.
 2. Agar:
11
Characteristics ofagar:
Long chain polysaccharide.
Inorganic salts,
A small amount of protein like material
Long chain fatty acids which are inhibitory to growth.
The minerals present are mainly magnesium and calcium.
A concentration of 1-2 percent usually yields a suitable gel.
T he jellifying property varies in different brands of agar.
12
Not add to the nutritive properties

Melting and solidifying points

 3. Peptone:
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Characteristics of peptone: • Act as buffer • Hygroscopic in
nature • Partially digested proteins obtained from meat,
heart muscle, casein, fibrin, soya meal, etc.

Digested proteins.
The important constituents are
Peptones,
Proteoses,
Amino acids,
Inorganic salts (phosphates, potassium and
magnesium, nicotinic acid and riboflavin).
14
4. Meat extract:
Prepared from fresh lean meat • Contains gelatin, proteoses, amino acids,
creatin, creatinine, purine, minieral salts, carbohydrates and Vit. B group

It is also available commercially and is known as Lab

Lemco.

5. Yeastextract:
Prepared from cells of baker’s yeast or Saccharomyces • Contains carbohydrates,
amino acids, growth factor(vit. B group) • Used as Vit. Source

It contains a wide range of amino acids, growth factors

and inorganic salts.
15 6. Maltextract:
It consists mainly of maltose (about 50%), starch, dextrin
and glucose, and contains about 5 percent of proteins
and protein breakdown products.
16 7. Blood and serum:

These are used for enriching culture media.

Either human or animal blood can be used.

Usually 5-10 percent blood is used and the most usual

concentration is 10 percent.

Serum is used in certain media.

17 CLASSIFICATION OF MEDIA
Media h a v e b e e n classified in many ways

A. On the basis of
B. On the basis of
Phases of growth C . Special M e d ia
nutritional factor
M e d ia
• Liquid m e d i a • Simple m e d i a • Enriched m e d i a
• Solid m e d i a • Complex media • Enrichment media
• Semi m e d i a • Chemically • Selective media
defined or • Indicator m e d i a
semisynthetic • Differential media
media
• Anaerobic growth
media
18 A. Phases of Growth Media:

Growth media are used in either of two phases:

liquid(broth) absence of agar

Solid (agar). 2% agar.
Semisolid media 0.2-0.5% agar

In some instances (e.g. certain blood culture methods), a

biphasic medium that contains both a liquid and a solid phase is
used.
 1. Liquid (broth) media:
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Bacterial growth is indicated by a change in broth’s

appearance from clear to turbid.

Isolation of bacteria to prepare pure cultures extremely

difficult.

The original media used by Louis Pasteur were liquids such

as urine or meat broth.
20

Example of liquidmedia

Nutrient broth

fluid thioglycollate
broth
21 Disadvantages of liquid media
Profuse growth.
Organisms cannot be separated.

Costly

Uses of liquid media

For obtaining bacterial growth from blood or water when

large volumes have to be tested.

For preparing bulk cultures of antigens or vaccines.
22 2. Solid (agar) media:
Solid media are made by adding a solidifying agent to the
nutrients and water. Agarose is the most common solidifying
agent
On solid media, bacterial making identification easy.
23 B. Based on Nutritional Factors

1 . Simple media

2. Complex media

3. Chemically defined or Synthetic media.

24 1. Simple media (Basal media)

Simple media are those which contain only basic nutrients

(peptone, yeast extract and glucose etc.) required for the
growth of ordinary organisms.

These simple media are generally used as the basis to

prepare enriched media; hence known as basal media.
 Example of simple media
25

Peptone water
26 2. Complex media:
Media that contain some ingredients of unknown chemical
composition.

Contains a variety of ingredients.

The exact chemical composition of these ingredients can be

highly variable although a specific amount of each
ingredient is in the medium.
27 Example of complex media

Luria broth
28 3. Chemically defined or Synthetic media:

Prepared from pure chemical substances and their exact

composition is known.

Chemically defined media are used for various experimental

purposes.

Special studies such as metabolic requirements of the

experimental microorganisms.
29 Example of synthetic media
Glucose salts Agar
30 References
https://microbeonline.com/types-of-bacteriological-culture-medium/
Kumar, S. (2012). Textbook of microbiology. JP Medical Ltd.