Bacteriological Culture

Media

Dr. Mejbah Uddin

Ahmed
 Several Terms
Media: The nutrient material in which
bacteria grow.
Culture: A bacterial growth in media.
Broth: A liquid medium.
Agar plate: Round culture dish with semi-
solid nutrient media.
 Several Terms
Slant: Tube with semi-solid media, with a
slanted surface.
Colony: A circular area of limited bacterial
growth.
Pure colony: A colony thought to contain
only the descendants of a single bacterium
 Several Terms
Fastidious: Bacteria that require very strict
nutrients.
Facultative: Able to live in various
conditions.
Obligate: Can only survive under certain
circumstances.
Aerobe: Bacteria that live on oxygen.
Anaerobe: Bacteria that are inhibited by
oxygen.
 Method of aenerobiasis
Exclusion of air:
● Liquid media may be heated to drive off
oxygen, cooled then inoculated & covered
with vaselin.
● Shake culture
● Stab culture.
■ Addition of reducing substances:
Glucose, thioglycolic acid, ascorbic acid.
 Method of aenerobiasis
Removal of air:
■ Removal of oxygen and replace by
nitrogen & hydrogen: McIntosh and Filde,s
anaerobic jar.
■ Self contained anaerobic system.
 Several Terms
Microaerophilic: Bacteria that require low
levels of oxygen.
Aerotolerant: Bacteria that prefer
anaerobic conditions but can survive in
minimal oxygen.
Colony morphology: Shape, appearance,
size, color of a colony.
 Types of Media
On the basis of constituent:
● Basic Media
● Enriched media
● Selective media
● Indicator media
● Transport media
 Types of Media
Basic media: These are simple media, such
as: nutrient broth media, nutrient agar
media.
Enrichment media: Media which contains
enrichment substances such as: blood,
serum, peptone, yeast extract, vitamins etc.
e, g; blood agar, Chocolate agar media.
 Types of Media

Selective media: Media which contains

substances like; bile salt, chemicals, dyes,
antibiotics which inhibit growth of unwanted
organisms: TCBS agar.
Indicator media: Media which contains
substances to differentiate microorganisms:
MacConkey agar.
 Types of Media
Transport media: These media contains
substances to prevent growth of unwanted
organisms and used for survival of
pathogens when cannot be cultured
immediately: Cary-Blair media.
 Culture Media

• Broth (liquid) in
a tube

▪ Agar (solid) in a
petridish
 Culture Media

Tryptic Soy Broth

(TSB)

MacConkey Agar
 Types of Media
On the basis of consistency:
● Liquid media.
● Semisolid media (0.5% agar).
● Solid media (1-2% agar).
■ Solidifying agents: Gelatine, Agar, Egg,
Serum.
Colonies of Various Species
 Sterilization of culture media
Depends on type and constituents:
● Autoclaving
● Filtration
● Steaming at 100ºC

*** Sterility testing?

*** How to inoculate culture media?
Inoculation of media in petri dish: By
wire loop.
Inoculation of slope: By straight wire.
Inoculation of stab culture: By straight
wire.
Inoculation of fluid media: By straight
wire, Pasture pipette.