CULTURE MEDIA

Microbial Nutrition

Nutrition = Obtaining
Required Substances from
the Environment
Essential Nutrients Must be
Provided for an Organism to
Survive and Reproduce
BASIC NUTRITIONAL REQUIREMENTS

WATER.
CARBON SOURCE.
NITROGEN SOURCE.
INORGANIC SALTS.
Phosphates. Sulfates.
Sodium. Potassium.
Magnesium. Iron.
Manganese. Calcium.
BACTERIAL VITAMINS.
Thiamine riboflavin
nicotinic acid pyridoxine
folic acid vitamine B12.
GASEOUS REQUIREMENTS
OXYGEN
Obligate aerobes
Facultative anaerobes.
Microaerophillic.
Obligate anaerobes.

CARBONDIOXIDE:
Capnophillic.
The oxygen requirement of parasites (mainly
bacteria) reflects the mechanism used by those
particular parasites to satisfy their energy needs
in host.
 CULTURE MEDIA
BASIC INGREDIENTS
PROTEIN SOURCE – peptone (a complex mixture of partially
digested proteins).
- special brands – Neopeptone & proteose peptone
- digest broths can also be used.
- meat extract (Lab Lemco).
MINERAL SALT – sodium chloride.
AGAR
- Prepared for using solid media.
- Obtained from sea weeds.
- Long chain polysaccharides.
- Also contains varying amounts of inorganic salts and
small quantities of a protein – like substance.
- Hydrolised at high temperature at high acid or alkaline
pH.
- Melts at 98ºC and usually sets at 42ºC depending on
agar concentration.
- Approximately 2% agar is used to prepare solid media.
- Manufacture either in long shreds or in powder form.
WATER- source of hydrogen and oxygen
TYPES OF CULTURE MEDIA
BASED ON THE AGAR
CONCENTRATION (Consistency):
1. Solid media – 2% agar.
2. Liquid media – absence of agar.
3. Semi solid media – 0.2-0.5% agar
 TYPES OF CULTURE MEDIA
BASED ON THE INGREDIENTS:
1. Simple or basal medium.
2. Complex medium.
3. Synthetic or defined medium.
4. Semisynthetic or semidefined medium.
5. Special medium
a. enriched medium.
b. enrichment medium.
c. selective medium.
d. indicator or differential medium.
e. transport medium.
f. Anaerobic medium.
g. sugar medium.
SIMPLE OR BASAL MEDIUM
Example: Nutrient agar &
Nutrient broth.
Consists of peptone, meat
extract, sodium chloride and
water.
Used to isolate separate
colonies for studying
1.colony morphology.
2.pigmentation. NUTRIENT AGAR
3.biochemical identification
tests.
NONPIGMENTED COLONIES
RED PIGMENTED COLONIES

GREEN PIGMENTED COLONIES YELLOW PIGMENTED

COLONIES
 COMPLEX MEDIUM
Exact chemical composition of ingredients of culture
medium is not known.
Widely used for the cultivation of bacteria for
diagnostic purposes.
SYNTHETIC & SEMISYNTHETIC MEDIUM

SYNTHETIC OR DEFINED MEDIUM:

Prepared from pure chemical substances and
the exact composition of the medium is known.
Used for various special studies such as
metabolic requirements.

SEMISYNTHETIC OR SEMIDEFINED MEDIUM:

Composition is approximately known.
 SPECIAL MEDIUM
ENRICHED MEDIUM
Substances such as blood, egg, serum
are added to the basal medium.
Used to grow bacteria which are more
exacting in their nutritional needs.
EXAMPLE: CHOCOLATE AGAR
Blood agar – hemolytic properties.
Chocolate agar - to supply blood
factors like X factor (Hemin) and Y
factor (NAD)

BLOOD AGAR
GAMMA HEMOLYSIS BETA HEMOLYSIS
(clear zone around
colonies)
ALPHA HEMOLYSIS
WITH GREENISH
DISCOLORATION
(no clear zone around
colonies)
BETA HEMOLYSIS UNINOCULATED BLOOD AGAR
 SPECIAL MEDIUM
ENRICHMENT MEDIUM
Obtaining the desired bacteria in
more numbers from the mixed
culture.
Contains substances that
stimulates the desired bacteria
or has inhibitory substances that
inhibits the growth of unwanted
bacteria.
EXAMPLE: Selenite F broth.
Tetrathionate broth.
EOSIN METHYLENE BLUE – E.COLI
 SPECIAL MEDIUM
SELECTIVE MEDIUM
This media enables a greater number of
the required bacteria to form colonies than
the other bacteria.
EXAMPLE: LJ medium for the selective
isolation of M.tuberculosis.
Desoxy cholate agar for Salmonella and
Shigella
Xylose lysine deoxycholate agar for
Salmonella and Shigella.
 SPECIAL MEDIUM
DIFFERENTIAL MEDIUM
A medium which has substances
incorporated in it, enabling it to
bring out differing characteristics
of bacteria and thus helping to
distinguish between them.
EXAMPLE: MacConkey agar –
consists of peptone, lactose, agar,
neutral red and taurocholate –
differentiates between lactose
fermentors and non-laactose
fermentors.
 NON - LACTOSE
FERMENTERS

LACTOSE FERMENTERS
(PINK COLONIES)

LACTOSE
FERMENTERS
 MACCONKEY AGAR

LACTOSE NON-LACTOSE
FERMENTER FERMENTER

(PINK COLOUR) (PALE OR

COLORLESS)
 PINK COLOUR
PALE YELLOW
(TRANSPARENT)
COLOUR
(TRANSPARENT)
MAC CONKEY AGAR

DARK
RED
COLOUR
(OPAQUE)

NUTRIENT AGAR BLOOD AGAR

 SPECIAL MEDIUM
ANAEROBIC MEDIUM
Media used to grow anaerobic organisms.
EXAMPLE: Robertson’s cooked meat
medium – contains cooked meat
Thioglycollate broth – Thioglycollate
Medium may be overlayed with paraffin
wax to prevent oxygen entry
 THIOGLYCOLLATE BROTH
ROBERTSON COOKED
MEAT BROTH