Gayam, Glydenne Glaire P.

BS Medical Technology 3A
Culture Medium

Bacte MWF 5:30-8:30

August 19, 2015
Use

Preparation

Components

1. Nutrient

Basal Medium

Dissolve 100g agar in 1L DH20. Heat,

dispense, autoclave.

2. Acetate Agar

Differential Medium

3. Alkaline Peptone Water

Enrichment Medium

4. Bismuth Sulfite Agar

Selective Medium

Add components, except sodium

acetate buffer, to distilled/deionized
water and bring volume to 900.0mL.
Mix. Heat and bring to boiling.
Autoclave and cool. Aseptically add
100.0mL of sterile sodium acetate
buffer. Mix.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
thoroughly. Adjust pH to 9.0. Autoclave.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
and heat until boiling. Boil for 1 min. Do
not autoclave. Cool.

Protein
Sugar
Indicators
Peptone
Sugar
Meat extract
Yeast extract
Sodium acetate buffer

5. Blood Agar, Sheep

Differential Medium

Add components, except sheep blood,

to distilled/deionized water and bring
volume to 950.0mL. Mix. Heat and
bring to boiling. Autoclave and cool
Aseptically add 50.0mL of sterile sheep
blood. Mix. Pour into sterile Petri
dishes.

Peptone
NaCl
Agar
Bismuth sulphite
Casein
Animal tissue
Glucose
Na2HPO4
FeSO47H2O
Trypticase soy agar
Brucella agar, or beef heart
infusion with 5% sheep blood

6. Phenylethyl Alcohol Agar

Selective Medium

7. Buffered Charcoal-Yeast
Extract Agar

Enrichment Medium

8. Chocolate Agar

Enrichment medium

9. Eoisin-Methylene Blue
Agar

Selective and Differential medium

Add components to distilled/deionized

water and bring volume to 1.0L. Mix,
gently heat and bring to boiling.
Distribute into tubes or flasks.
Autoclave.
Pour into sterile Petri dishes or leave in
tubes.
Add components, except L-cysteine
solution, to distilled/deionized water and
bring volume to 1.0L. Mix, heat and
bring to boil for 1 min. Autoclave and
cool. Add 4.0mL of L-cysteine solution.
Mix and pour into sterile Petri dishes
with constant agitation to keep charcoal
in suspension.
Add components, except supplement B
solution and sheep blood, to
distilled/deionized water and bring
volume
to 890.0mL. Mix and heat until boiling.
Autoclave and cool. Aseptically
add 100.0mL of sterile, defibrinated
sheep blood. Heat while stirring and
bring to 85C for 510 min. Cool to
50C. Aseptically add 10.0mL of sterile
supplement B. Mix and pour into sterile
Petridishes or distribute into sterile
tubes.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix,
heat and bring to boiling. Distribute into
tubes or flasks. Autoclave and pour into

Agar
Tryptose
NaCl
Beef Extract
Phenylethyl alcohol

Yeast extract
Agar
Charcoal
salts

Peptone base, enriched with

solution of 2% hemoglobin or
isovitalex (BBL)
NaCl
Cornstarch
Sheep blood

Peptone base (lactose)

Eosin Y
Methylene Blue indicator
Sugar

sterile Petri dishes.

10. Hydrogen Sulfide, Lead
Acetate

Differential culture

11. Kliglers Iron Agar

Differential medium
Identify if a gram-negative rod is a
glucose or lactose fermenter or
both

12. Lim Broth

Enrichment broth
Isolate S. agalactiae

13. Lysine-Iron Sugar

Differential medium
Identify species of
Enterobacteriaceae

14. MacConkey Agar

Selective and Differential medium

15. Mannitol Salt Agar

Selective and Differential medium

Add components to distilled/deionized

water and bring volume to 1.0L. Mix,
heat and bring to boiling. Distribute into
tubes or flasks. Autoclave and pour into
sterile Petri dishes or leave in tubes.
Allow tubes to cool in a slanted
position.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix,
heat and bring to boiling. Distribute into
tubes. Autoclave. Pour into sterile Petri
dishes or leave in tubes.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
thoroughly. Distribute into tubes or
flasks. Autoclave.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
then heat while stirring and bring to
boiling. Distribute into tubes in 10.0mL
volumes. Autoclave. Allow tubes to cool
in a slanted position.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
then heat while stirring until boiling.
Autoclave. Pour into sterile Petri dishes
or distribute into sterile tubes.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix,
heat while stirring and bring to boiling.
Distribute into tubes or flasks.

Lead acetate
Agar
Peptone
Glucose

Sugars
Indicator
Peptone
Sulfur source
Peptones, yeast extract,
dextrose

Amino acid
Carbohydrate
Indicators
Sulfur

Peptone base
Indicator
NaCl
Bile Salts
Peptone base
Mannitol
Indicator
Beef extract

Autoclave. Pour into sterile Petri dishes

or leave in tubes.
16. Mueller-Hinton Agar

Transparent medium

17. New York City Medium

Selective medium
Isolate N. gonorrhoea and N.
meningitidis from specimens
containing mixed normal flora

18. Selenite Broth

Enrichment Broth

19. Triple-Sugar Iron Agar

Differential medium
Identification of Glucosefermenters from Non-glucosefermenters gram negative rods

20. Trypticase Soy Agar

All-purpose/ Basal medium

Add components to distilled/deionized

water and bring to 1.0L. Mix, heat and
bring to boiling. Distribute into tubes or
flasks. Autoclave. Pour into sterile Petri
dishes or leave in tubes.
Have all solutions prepared and at 45
50C. Aseptically combine components.
Mix. Pour into sterile Petri dishes.

Add components to distilled/deionized

water and bring volume to 1.0L. Mix,
heat and bring to boiling. Do not
autoclave. Distribute into sterile tubes in
10.0mL volumes.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix,
heat and bring to boiling. Distribute into
tubes or flasks. Autoclave and cool in a
slanted position to form a
1.0-inch butt.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix,
heat and bring to boiling. Distribute into
tubes or flasks. Autoclave. Do not
overheat. Pour into sterile Petri dishes
or leave in tubes.

NaCl
Casein
Animal Tissue
Animal infusion
Casein extract
Starch
Agar
Peptone agar base with
cornstarch
Horse blood cells
Horse plasma, citrated
Yeast dialysate
Peptone base broth
Sodium biselenite

Sugars
Indicator
Peptone
Sulfur source

Peptone
Casein
Sodium chloride

21. Trypticase Soy Broth

All-purpose medium/ Enrichment

medium

Add components to distilled/deionized

water and bring volume to 1.0L. Mix,
heat and bring to boiling. Distribute into
tubes or flasks. Autoclave then mix
thoroughly.

Mahon, C.R. et. al (2015). Textbook of Diagnostic Microbiology (4th edition). Saunders Elsevier.
Tille, P.M. (2014). Bailey & Scotts Diagnostic Microbiology (13th edition). Elsevier Mosby.
Atlas, R.M. (2010). Handbook of Microbiological Media (4th edition). CRC Press.

Soybean
Casein
NaCl
Glucose
Dipotassium phosphate