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CULTURE MEDIA
Brief History

bread, potato, coagulated egg albumin, starch

paste, and meat
few pathogenic organisms would grow on
potato, and other media such as egg,
prevented adequate inspection of the colonies
for growth.
Bartolemeo Bizio, 1832
black spots on communion wafers
Other breads: Serratia marcescens
 Joseph Lister
Transactions of the
Pathological Society of
Londonfor the session
187778
Lactic acid fermentation
in milk
limiting dilution
method
Bacillus lactis
Penicillium glaucum,
Aspergillus glaucus, and
two forms ofMucor
 Robert Koch
1881
Father of Culture
Media
Beef broth or
aqueous humor
Int Medical
Congress -Nutrient
Gelatin
Liquified @ 25C
Gelatinase is present
in most proteolytic
bacteria
 Fanny Hesse
1881, Laboratory
technician and
illustrator
Walther Hesse
Suggested the use
of agar
Was never credited
for her contribution
until 1939
 Julius Richard Petri
Flat plate covered
with bell jar
1887
 Friedrich Loeffler
Meat extract broth
Lacks amino-
nitrogen for optimal
growth
Corynebacterium
diphtheriae
Peptone
Sodium chloride
1. Providing substance bacterial growth
Carbohydrates, peptones, proteins,
vitamins,etc
2. Indicators
3. Dyes and other substances
Which inhibit undesired bacteria
4. Organic and inorganic activity
For the maintenance of constant pH
 CLASSIFICATION OF BACTERIAL
CULTURE
ACCORDING TO
THEIR PHYSICAL
STATE
SOLID MEDIA
Agar
2-3%; 38C
Gelatin
1-1.5%; below
25C
Albumin
Loefflers Blood
Serum
 SEMI-SOLID
Cultivation of
microaerophiles and
determination of
motility
LIQUID /BROTH
propagation of large
number of
organisms,
fermentation studies,
and various other
test
 ACCORDING TO THEIR
COMPOSITION
SYNTHETIC MEDIA
chemically defined
Purified ingredients
Ringers solution
Lockes solution
 ACCORDING TO THEIR
COMPOSITION
NON-SYNTHETIC
MEDIA
chemically
Undefined
Composition
unknown
Meat extract broth
Vitamin broth
Nutrient Broth
 ACCORDING TO THEIR
COMPOSITION
LIVING TISSUE MEDIA
Living tissues
Viruses and
Rickettsiae
HeLa tissue culture
cells
Tissue-plasma
roller cells
Embryonated egg
 SIMPLE MEDIA
Primary isolation

Peptone water
Nutrient agar/broth
 ENRICHED MEDIA
Blood Agar
Chocolate agar
Loefflers blood
serum slant
Lowenstein-Jensen
Medium
DIFFERENTIAL MEDIA

Dyes, sugars, indicators to elicit a

biochemical reaction
Antibiotics
Color reaction
Fermentation studies
 DIFFERENTIAL MILDLY SELECTIVE
MEDIA
Eosin Methylene
Blue Agar
MacConkey Agar
 DIFFERENTIAL MODERATELY
SELECTIVE MEDIA
Salmonella Shigella
Agar
Xylose Lysine
Deoxycholate Agar
Deoxycholate
Citrate Agar
Hektoen Enteric
Agar
 HIGHLY SELECTIVE MEDIA
Promotes the
growth of a specific
organism
Bismuth sulfite
agar
Lowenstein-Jensen
Agar
Campy Blood Agar
Fletchers Medium
 TRANSPORT MEDIA
used when
specimen cannot
be cultured soon
after collection
Cary-Blair medium
Amies medium
Stuart medium
 ENRICHMENT MEDIA
Designed for stool
cultures where the
normal flora are
suppressed favoring
the growth of
enteropathogens
Selenite F broth,
tetrathionate, GN
broth, alkaline
peptone broth,
Campy-Thio
 NUTRIENT AGAR/BROTH
ox heart, water, peptone,
sodium chloride
2.5% agar
pH 7.3.
basal media for the
preparation of other
media
To study soluble products
of bacteria.

.
 PEPTONE WATER
Peptone 1% and
sodium chloride
0.5%.
It is used as base
for sugar media
and to test indole
formation.
 BLOOD AGAR
Most commonly
used medium.
TSA with 5-10%
defibrinated sheep
or horse blood.
Both an
enrichment and
indicator.
hemolysis
 Chocolate agar
Heat-lysed blood
Nicotinamide
adenide
dinucleotide (factor
V) hemin (factor X)
culture of
pneumococcus,
gonococcus,
meningococcus
and Haemophilus
 MacCONKEY AGAR
Enterobacteriaceae
agar, peptone,
sodium chloride, bile
salt, lactose and
neutral red
bile salt inhibits
growth of many other
bacteria.
Lactose fermenters
turn neutral red to
pink
MUELLER HINTON INFUSION AGAR

Disc diffusion
sensitivity tests for
antimicrobial drugs
 HISS SERUM WATER MEDIUM
fermentation
reactions of bacteria
which can not grow
in peptone water
sugar media
CTSA + serum +
sugars
pneumococcus,
Neisseria,
Corynebacterium.
 LOWENSTEIN-JENSEN MEDIUM
Egg stimulates the
growth of tubercle
bacilli
Malachite green
inhibits growth of
organisms
Glycerol promotes the
growth of
Mycobacterium
tuberculosis
M.bovis omit glycerol,
add Na pyruvate
 DUBOS MEDIUM
drug sensitivity of
tubercle bacilli
casein hydrolysate,
asparagin and salts
Tween 80 causes
dispersed growth
and bovine
albumin causes
rapid growth.
 LOFFLERS SERUM
C. diphtheriae (6
hours)
rapid diagnosisof
diphtheria
demonstrate
volutin granules
sheep, ox or horse
serum.
 TELLURITE BLOOD AGAR
selective medium
forCorynebacteriu
m diphtheriae
Tellurite
inhibitor and
indicator
produce black
colonies.
 EOSIN METHYLENE BLUE
selective and
differential for enteric
Gram-negative rods.
Lactose-fermenting
colonies are coloured
and nonlactose-
fermenting colonies
are nonpigmented.
Selects against gram
positive bacteria.
 XYLOSE LYSINE
DEOXYCHOLATE
Salmonella and
Shigella species from
stool specimens.
Na DC-inhibits G(+)
Xylose-not oxidized
by Shigella
Lysine cadaverine
by Salmonella
sodium
thiosulfateandferri
c ammonium citrate
 SALMONELLA SHIGELLA
AGAR
Selective
Modification of Desoxycholate
Citrate Agar
Bile Salts,Sodium
CitrateandBrilliant
Greenserve to inhibit gram-
positive, coliform organisms and
Proteus
Lactoseis the carbohydrate
present inSalmonella
ShigellaAgar
ThiosulfateandFerric Citrate -
black centered colonies.
Neutral redturns red in the
presence of an acidic pH, thus
showing fermentation has
occurred.
 DEOXYCHOLATE CITRATE
AGAR
Differential selective
SS
Na deoxycholate
and Sodium
citrate
Inhibits G(+)
bacteria
Proteus
lactose and
neutral red
 TETRATHIONATE BROTH
Selective for SS
Peptone based
broth
Bile salts and Na
thioSO4 inhibit GP
organisms and
most
Enterobacteriaceae
 SELENITE F BROTH
Selective enrichment for SS
Casein enzymic
hydrolysate
Lactose
Selenite
Sodium
phosphatemaintains a
stable pH and also lessens
the toxicity of selenite.
Sodium seleniteinhibits
many species of gram-
positive and gram-
negativebacteria including
enterococci and coliforms.
 THIOSULFATE-CITRATE-BILE-
SUCROSE (TBCS) AGAR
selective isolation ofVibrio
choleraeand other
enteropathogenic vibrios.
Thiosulfateandsodium
citrate, as well inhibit the
growth of Enterobacteria.
Ox bileandsodium cholate
Bromthymol
BlueandThymol Blueare
pHindicators
Thiosulfate and ferric
sulfate
Yeast extractandpepton
Sodium chloride
CHARCOAL YEAST AGAR (BCYA)
Legionella
pneumophila
Yeast extract
L-Cysteine HCl
Charcoal activated :
decomposes hydrogen
peroxide
Ferric Pyrophosphate;
iron source
ACES buffer
: to stimulate growth
 CAMPYLOBACTER MEDIUM
Selective for C. jejuni and
C. coli
Peptones, dextrose, yeast
extract, and blood
Vancomycin inhibits
gram-positive bacteria
polymyxin B inhibits
most gram-negative
bacilli except forProteus.
Trimethoprim is
inhibitory forProteusspp.
Nystatin is an antifungal
agent.
 CARY-BLAIR MEDIUM
Low nutrient
content, low
oxidation-reduction
potential
Transport of fecal
specimens
Salmonella,
Shigella, Vibrio or
Campylobacter
AMIES TRANSPORT MEDIUM
Na thoiglycollate
Maintenance of
organisms in
original specimen
Neisseria
gonorrhoeae
PEPTONE WATER SUGAR MEDIUM

Sugar fermentation
lactose, glucose,
mannitol
Andrades indicator
Inverted Durham
tube collects gas
Color change =
acid
 MOTILITY- INDOLE-UREA
used to
differentiate
enterobacteria
species by their
motility, urease,
and indole
reactions.
TRIPLE SUGAR IRON AGAR
 KlIGLER IRON AGAR

Same as TSI
No sucrose
 CHRISTENSENS UREA MEDIUM
used to identify
urea splitting
organisms, e.g.
Proteus.
A purple pink
colour indicates
urea splitting
 BORDET-GENGOU MEDIUM
Bordetella
pertussis (Hg drop
colonies)
Increased
concentration of blood
allows growth
agar, potato,
sodium chloride,
glycerol, peptone
50% horse blood
Penicillin
SABOURAUDS DEXTROSE AGAR
enzymatic
digest of casein
Dextrose
hloramphenicol
and/or tetracycline
Gentamicin
pH 5.6
MODIFIED THAYER MARTIN MEDIUM

BA base enriched
with hemoglobin
and supplements
Colistin, nystatin,
vancomycin and
trimethoprim
N. gonorrhoeae
and N. meningitidis
bye
 PREPARATION OF THE MEDIA
Use sterile
instruments,
glassware etc to be
used or autoclave
them beforehand.
 PREPARATION OF THE MEDIA
Measure out the
materials ( agar
base, water)
CHARCOAL YEAST AGAR
 PREPARATION OF THE MEDIA
Dissolve agar
powder completely
and cover openings
before placing in
water bath.
 PREPARATION OF THE MEDIA
Autoclave the
solution
For slants or tubes:
transfer solution to
tubes prior to
autoclaving.
 PREPARATION OF THE MEDIA
Take out of
autoclave, pour into
plates (20ml).
Place the tubes at
an angle to set into
slants.
For blood agar, add
sheep or horse blood
to slightly cooled
solution (50C) prior
to pouring.
 PREPARATION OF THE MEDIA
For chocolate agar:
add sheep/horse
blood, heat-lyse
the solution at
56C water bath

Once set, place

them in sealed
sterile bags and
store at 4C.
PREPARATION OF THE MEDIA
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