Culture Media

CULTURE MEDIA
HAFIZ AHMAD LUBAB KTK

INSTITUTE OF NURSING SCIENCES
KHYBER MEDICAL UNIVERSITY
Unit Objectives
Perform the procedure to prepare slides of culture

media.
Follow the professional norms to prepare slides of

culture media.
Culture/Medium Defined
• 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.
• Inoculation: Introduction of microbes into the

medium.
Reasons for Bacterial Cultural
• Subsequent clinical diagnosis.
• Studying its morphology and its identification.
• 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.
The Requirements for Bacterial
Growth
Physical Requirements:
• Temperature
• pH
• Osmotic Pressure
The Requirements for Bacterial
Growth
Chemical Requirements:
• Carbon / Nitrogen
• Phosphorous
• Oxygen
• Hydrogen
• Trace Elements
Types of Media
Chemically Defined Media: A chemically defined

medium is a growth medium suitable for the in vitro cell
culture of human. Exact chemical composition is
known.e.g Basal media
Complex Media: These are media containing nutrients

is unknown quantities that are added to bring about a
particular characteristic of a microbial strain.e.g B agar.
Types of Media
 A Chemically Defined Medium for Growing a Typical

Chemoheterotrophic, such as E.coli.
Constituent Amount
Glucose 5.0g
Ammonium phosphate 1.0g
Sodium Chloride (NaCl) 5.0g
Magnesium Sulfate 0.2g
Potassium Phosphate 1.0g
Water 1 liter
Types of Media
 Composition of Nutrients, a Complex Medium for the Growth

of Heterotrophic Bacteria
Constituent Amount
Peptone (P digested protein) 5.0g
Beef extract 3.0g
Sodium Chloride 8.0g
Agar 15.0g
Water 1 liter
Types of Culture Media
 Selective Media:
• Suppress unwanted microbes and encourage desired
microbes.
 Differential Media:
• Make it easy to distinguish colonies of different
microbes.
 Enrichment Media:
• Encourages growth of desired microbes.
Examples of Selective Media
• Blood agar
• MacConkey agar
• Hektoen enteric agar (HE)
• Mannitol salt agar (MSA)

Examples of Differential
Media
• Eosin methylene blue (EMB)
• MacConkey agar
• Mannitol salt agar (MSA)

Examples of Enriched Media
• Chocolate agar
• Blood agar
Blood Agar
• Contains 5-10% mammalian blood (usually sheep or horse)
• Contains meat extract, NaCl, and agar
• Used to detect hemolytic activity
• Alpha-hemolysis- partially lysis of RBC(Streptococcus P)
• B-hemolysis- Complete lysis of RBC (E.Coli)
• Y-hemolysis- no lysis ( Staplococcus Epidemidis)

Blood Agar
•
MacConkey Agar
• It contains bile salts (to inhibit Gram-positive bacteria)
• Crystal violet dye
• Neutral red dye (which stains microbes fermenting

lactose)
• Lactose
• Peptones
MacConkey Agar
Mannitol Salt Agar (MSA)
• It contains a high concentration (7.5-10%) of salt

(NaCl), making it selective for Staphylococci and
Micrococcaceae.
Mannitol Salt Agar (MSA)
Eosin Methylene Blue
• Is a selective stain for Gram-negative bacteria
• Inhibits the growth of Gram-positive bacteria and

provides a color indicator distinguishing between
organisms that ferment lactose (e.g., E.coli) and those
that do not (e.g., Salmonella, Shigella).
Eosin Methylene Blue
Chocolate Agar
• Is a non-selective, enriched growth medium.
• Contains red blood cells, which have been lysed by

heating.
• It is a variant of the blood agar plate, containing red

blood cells that have been lysed by slowly heating to
80°C.
Chocolate Agar
Lowenstein-Jensen medium
• For the cultivation and differentiation of

Mycobacterium species.
Mueller-Hinton Agar (MHA)
• For antimicrobial susceptibility testing.

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CULTURE MEDIA

HAFIZ AHMAD LUBAB KTK

Perform the procedure to prepare slides of culture

Follow the professional norms to prepare slides of

• CULTURE : Is the term given to microorganisms that

• MEDIUM: Is the term given to the combination of ingredients

• Inoculation: Introduction of microbes into the

• Subsequent clinical diagnosis.

• Studying its morphology and its identification.

• Bacteria have to be cultured in order to obtain antigens

• Certain genetic studies and manipulations of the cells

Chemically Defined Media: A chemically defined

Complex Media: These are media containing nutrients

 A Chemically Defined Medium for Growing a Typical

 Composition of Nutrients, a Complex Medium for the Growth

• Hektoen enteric agar (HE)

• Mannitol salt agar (MSA)

• Mannitol salt agar (MSA)

• Contains 5-10% mammalian blood (usually sheep or horse)

• Contains meat extract, NaCl, and agar

• Used to detect hemolytic activity

• Alpha-hemolysis- partially lysis of RBC(Streptococcus P)

• B-hemolysis- Complete lysis of RBC (E.Coli)

• Y-hemolysis- no lysis ( Staplococcus Epidemidis)

• It contains bile salts (to inhibit Gram-positive bacteria)

• Crystal violet dye

• Neutral red dye (which stains microbes fermenting

• It contains a high concentration (7.5-10%) of salt

• Is a selective stain for Gram-negative bacteria

• Inhibits the growth of Gram-positive bacteria and

• Is a non-selective, enriched growth medium.

• Contains red blood cells, which have been lysed by

• It is a variant of the blood agar plate, containing red

• For the cultivation and differentiation of

• For antimicrobial susceptibility testing.

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