AGAR AGAR

Identification : Powder or dried shreds

Light straw coloure
Properties :Prepared from sea weed a long chain
poly sacharide, melts at 95°C & solidifies on
cooling to 42 C.

Use
Solifying agent to prepare solid media
Concentrations used
Solid media 2% Japanese agar
1.5% Newzealand agar
Semi solid agar 0.2 0.5%
Hard agar - 4-6% ( to prevent swarming growth of Proteus &
CI. tetani)
Agarose (purified agar) 1% for gel
-

electrophoresis
 NUTRIENT BROTH
IDENTIFICATION :
Straw colored liquid medium with label NB
TYPES :1. Meat infusion
broth
2. Meat extract broth
3.Digest broth

CONSTITUENTS :
Lean meat/Meat extract
Peptone
NaCl
Water, Ph7.4

STERILISATION: Autoclaving at 121°c for 15 min.

USES Basal liquid medium
Basis of most media.
 PEPTONE WATER

medium with
I D E N T I F I C A T I O N : Colourless transparent
Lable PW.

CONSTITUENTS : Peptone
Sodium Chloride
Distilled water

TYPE OF
Basal liquid medium.
MEDIUM

media
1.Base for carbohydrate fermentation
USES 2.Indole test.
3.Demonstration of motility
 PEPTONE

DENTIFICATION: Granular powder

Golden yellow coloured.

Animal proteins (lean meat, heart muscle)

SOURCE
Vegetable proteins (soya flour)
by digestion with proteolytic enzymes

CONSTITUENTS: Peptone, Proteoses ,Aminoacids

Inorganic salts , minerals( like K, Mg)
Accessory growth factors like Nicotinic acid,
Riboflavine)

USES :Base for preparation of different media

Peptone water
 GLUCOSE BROTH
IDENTIFICATION: Straw coloured Broth (50
ml)
Big (4 oz.) mediçal flat bottle
(screw capped)
CONSTITUENTS: Meat extract broth
Sodium chloride
Peptone
Glucose (1%)
Anticoagulant-Citrate/ Liquoid (Sodium polyanethol sulphonate)
STERLISATION: -

Autoclving at 116'C 10 lbs. x 10 minutes

-Filtration
-Tyndallisation
USES: Blood culture
*Promotes luxuriant growth of bacteria
Anaerobic medium (glucose as
reducing agent)
 NUTRIENT AGAR
IDENTIFICATION Solid plate Medium
Straw Yellow coloured
COMPOSITION Transluscent
Peptone Beaf Extract,
,

Agar Distilled water

,

TYPE OF MEDIUM pH- 6.8

USES Basal solid Medium
Basal medium for other
For media
Antimierobial Susceptibility
Pigments better observed Testing
Colonies for Antisera
In semi solid state:
For Catálase test. agglutination test
Preservation of bacteria
Holding medium for transport
Motility testing
 MAC CONKEY AGAR
IDENTIFICATION Solid plate medium, Reddish brown colour,
Transluscent
CONSTITUENTS Peptone, Lactose, Bile salt - selective agent,

Agar, Neutral red (2%) indicator, Distilled water,

pH-7.4
STERILISATION Autoclaving at 121°C x 15 mins.
PRINCIPLE Laçtose
Bacterial fermentation
Acid
Neutral red Pink colour
RESULT Pink colony Lactose fermenter
Pale or colourless colony- Lactose nonfermenter.
TYPE OF MEDIUM: Seleçtive I Differential/ Indicator
USES Primary selective medium for Enterobacteriaceae.
 MILK AGAR
Identification : Solid plate medium
White coloured opaque

Constituents Sterile fresh milk added

nutrient to molten sterile

agar at 56 0 C.
Type of medium
-Enriched medium
Uses To enhance pigment production of
Staphylococci
Golden yellow - Staph. aureus
White Staph. albus
Lemon yellow Staph.citreus
Modification Salt milk agar (7- !0% ) selective
Staphylococcus aureus
medium for
IDENTIFICATION Solid
BLOOD AGAR
plate medium, Blood red colour,
Opaque
CONSTITUENTS Nutrient agar, Sterile
sheep blood 5% 10% -

(Horse, Rabbit, Human blood may be used),

STERILISATION :
pH-7.2-7.4
Autoclaving Nutrient agar at 121Cx 15 mins. 15lbs
Cooled 55°C
and blood added
to
TYPE OF MEDIUM:Enriched medium aseptically.
USES and Differential medium (Haemolysis)
Primary isolation medium for Staphylococci.,
*Growth of fastidious organisms. Streptococci & Y easts.
Haemolysis pattern observed.
*Swarming of Proteus
*
Sensitivity testing of Streptococci.
MODIFICATIONS Selective Blood agar -Addition of Antibiotics.
Potassiumtellurite blood agar- C.
diphtheriae (Black colonies)
Hard Blood agar ( 6% agar) Prevents
swarming of Proteus.
-
 CYSTEINE LACTOSE ELECTROLYTE
DEFICIENT (CLED) MEDIUM
IDENTIFICATION SOLID PLATED
MEDIUM
TRANSLUSCENT
LIGHT S COLOUR
INGREDIENTS-PEPTONE, T RYPTONE,
NE,TRV

L-CYSTEINE,B LABLEMCO,
ROMOTHYMOL
BLUE,AGAR,WATER
OLOUR

INDICATOR USED-
BROMOTHYMOL BLUE
STERILISATION- AUTOCLAVING AT
FOR 15 MINS. 121 'C
TYPE OF MEDIUM DIFFERENTIAL
-

MEDIUM
USES- DIFFERENTIATES
LACTOSE
NON FERMENTING COLONIES.
FERMENTING&
INHIBITS SWARMING OF
PROTEUS AS
ELECTROLYTE IS DEFICIENT.
SPECIFICALLY USED FOR
URINARY PATHOGENS ISOLATION OF
SUPPORTS THE GROWTH
OF
STAPHYLOCOCCI,STREPTOCOCCI,
THAT FAIL TO
GROW IN CANDI
MACCONKEY.
DA
 CHOCLATE
(Syn HEATED
AGAR
BLOOD
-

AGAR)

IDENTIFICATION: Solid phase medium

Chocolate coloured
Opaque
PREPARATION Sterilised _olid blood
till chocolate brown. agar heated to 75°c

USES : Isolation of
fastidious organisms like
-H.influenzae
-Neisseriae
-Stepto pneumonae
 SABOURAUD'S DEXTROSEAGAR (SDA)
IDENTIFICATION Read label
Straw yellow coloured slant in tube with label SDA

CONSTITUENTS Dextrose (4%) High glucose content

-

Peptone, Agar, Distilled water, pH-5.6

STERILISATION Autoclaving at 121°Cx 15 mins.

TYPE OF MEDIUM Selective medium

USES Primary isolation medium for fungi.

MODIFICATIONS SDA with antibiotics,

*
Mycosel agar (SDA+ Antibiotics +Cycloheximide)
Cycloheximide prevents growth of saprophytic fungi.
 ROBERTSON'S COOKED MEAT BROTH
IDENTIFICATION Meat particles atthe bottom,
Pale yellowish brown liquid.

CONSTITUENTS Chopped & cooked

meat (fat free)
Peptone Infusion Broth, pH -7.5
STERILISATION Autoclaving at 121°C x 15 mins.
USES *
Cultivation of anaerobes.
*Preservation of stock cultures of aerobic
*Recovery medium for spores. organisms.
*Saccharolytic and Proteolytic activity of
Clostridium species.
MODIFICATIONS Salt cooked meat broth-
for Staphylococcu.
Enrichment medium
 PLOO DD CULTURE BOTTLE
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SPECImENI Te STED
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Blood, v a l lut'e
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L Lasouk
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APVANTAGE MoAL enel
 CASTANEDA'S BIPHASIC
MEDIUM
IDENTIFICATION: 100 mlemedical flat bottle having solid slant
& liquid medium(Biphasic)

CONSTITUENTS :Solid phase-Sterile serum glucose agar

Liquid phase-Sterile serum glucose broth
PH7.2

STERILISATION :Autoclaving at 110°c x 10 min

Sterile broth added after slant preparation

USES Primary isolation media for Brucella

&Salmonella.
Cultivation of blood for isolation of slow growing
bacteria

ADVANTAGES Allows subculture on solid phase without opening

the bottle.
Avoids contamination associated with frequent
subculturing.
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