C ULTURE M EDIA S LIDES

M icrobiology
BSN Semester: I
Unit:8
IFTIKHAR UL HAQ
DEMONSTRATOR
INS-KMU, PESHAWAR
 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: Exact chemical composition is known.

 Complex Media: Extracts and digests of yeasts, meat, or plants.

 Types of Media

A Chemically Defined Medium for Growing a Typical Chemoheterotropc, such as

E.coli
Constituent Amount
Glucose 5.0g
Ammonium phosphate, (NH4H2PO4) 1.0g

Sodium Chloride (NaCl) 5.0g

Magnesium Sulfate (MgSO4 - 7H20) 0.2g
Potassium Phosphate, (K2HPO4) 1.0g
Water 1 liter
 Types of Media

Composition of Nutrients, a Complex Medium for the Growth of Heterotrophic

Bacteria
Constituent Amount
Peptone (Partially 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

• B-hemolysis- Complete lysis of RBC

• Y-hemolysis- no lysis
 MacConkey Agar

• It contains bile salts (to inhibit

Gram-positive bacteria)

• Crystal violet dye

• Neutral red dye (which stains

microbes fermenting lactose)

• Lactose

• Peptones
 Mannitol Salt Agar (MSA)

• It contains a high concentration (7.5-

10%) of salt (NaCl), making it selective
for Staphylococci and Micrococcaceae
 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).
 Chocolate Agar

• Is a non-selective, enriched growth medium

• Contains red blood cells, which have been lysed by heating

 Lowenstein-Jensen medium

• For the cultivation and differentiation of Mycobacterium species

 Mueller-Hinton Agar (MHA)

• For antimicrobial susceptibility testing