CULTURE MEDIA

CANELLIE P. CANLAS
TERMINOLOGIES:

NOUN:
1. CULTURE
VERB:

2. INOCULATE/ PLANT/ CULTIVATE

introducing the microorganism to the culture media

3. TRANSPLANT/ SUBCULTURE
transfer of microorganisms from one CM to another
CLASSIFICATION OF CULTURE MEDIA

I. ACCORDING TO COMPOSITION
A.
all ofSYNTHETIC/ CHEMICALLY DEFINED
the components are known

B. NON-SYNTHETIC/ NON-CHEMICALLY DEFINED/

one of the components are not defined by chemical formula, making the CM complex
COMPLEX
C. TISSUE CULTURE MEDIA
1. VERO CELL
2. Mc COY CELL LINE
3. CHICKEN EMBRYO
4. A549 CELL LINE
5. HEP-2 CELL LINE
• II. ACCORDING TO PHYSICAL STATE/ CONSISTENCY
• A. LIQUID no agar bec, it doesnt have solidifying agent

» Agar- solidifying agent of choice in the preparation of culture media

from red algae(sea weeds) of phylum rhodophyta w/ a genus gelidium

• B. SEMI-SOLID
1. SULFIDE INDOLE MOTILITY MEDIUM
C. SOLID -
a. liquefiable- artificially prepared in the laboratory

b. non-liquefiable- solid CM that can't be deliquify

rice grains and potato slices -- fungi
cooked meat -- anaerobic organisms
• ACCORDING TO MANNER
OF DISPENSING/
FORMATION
• A. PLATED

• B. TUBED
butt, slant, butt slant

used in•the
C.isolation
BOTTLED of Airborne microorganisms in the blood sample
» A. CASTANEDA
biphasic medium, usually used in the culture o blood
Brucella
» B. SEPTI-CHEK
used in the laboratory in the present, innovation of castaneda bollte

» C. BACTEC
• IV. ACCORDING TO FUNCTION/ USE
– A. GENERAL PURPOSE/ BASIC/ BASAL/ SUPPORTIVE/
GENERAL ISOLATION CULTURE MEDIA
basically allows / supports the growth of almost all organism
– B. ENRICHED CULTURE MEDIA
– 1. BLOOD AGAR PLATE
– 2. CHOCOLATE AGAR PLATE
general purpose media w/ enriching substance/ additive, supports fastidious organism

– C. ENRICHMENT CM
– 1. ALKALINE PEPTONE WATER
– 2. SELENITE F BROTH
– 3. TETRATHIONATE BROTH
– 4. HAFNIA BROTH
– 5. GN BROTH
• D. TRANSPORT CULTURE MEDIA
– A. CARY-BLAIR
– B. AMIE
– C. STUART
– D. TRANSGROW

• SELECTIVE CULTURE MEDIA

FAVORS THE GROWTH OF THE ORGANSISM OF THE INTEREST AND INHIBITS THE REST

• INHIBITORS:
– 1. GRAM POSITIVE
• A. DYES
• B. BILE SALTS

– 2. GRAM NEGATIVE
• A. POTASSIUM TELLURITE - for Corynebacterium diptheriae
• B. SODIUM AZIDE
 DIFFERENTIAL CULTURE MEDIA

1. EOSIN METHYLENE BLUE (EMB) MEDIUM

SELECTIVE FOR:
INHIBITORS:

pH indicator :
 EMB

RLF:

Lactose Fermenter
LLF:
Lactose
Non-Lactose Fermenter
Differentiation of Family Enterobacteriaceae based on
Lactose fermentation
• RLF Escherichia, Enterobacter, Klebsiella

• LLF Hafnia, Serratia, Citrobacter

Salmonella arizonae
Shigella sonnei
Yersinia enterocolitica

• NLF ALL Salmonella except S. arizonae

ALL Shigella except S. sonnei
Proteus
Providencia
Morganella
Edwardsiella
Characteristic growth of Rapid Lactose
Fermenters in EMB
1. Escherichia coli large pink/ violet w/ greenish metallic sheen (fish eye colony)

2. Enterobacter large, pink or violet w/ or w/o dark center

3. Klebsiella large, pink or violet, mucoid colonies, w/ w/o dark center

2. Mc Conkey (MAC) Medium

• Light pink

• Selective for:

• Inhibitors:

• Differential
• pH Indicator:
 MAC

RLF:

Lactose Fermenter
LLF:
Lactose
Non-Lactose Fermenter
 3. SALMONELLA-SHIGELLA (SSA) MEDIUM

Light orange

Salmonella & Shigella

Selective for: Brillliant green
• Inhibitors: Bile salts
Sodium citrate
Differential
pH indicator: Neutral red
• Indicators:
H2S indicator: Ferric ammonium citrate

Sulfur source:Na thiosulfate

 RLF:

Lactose Fermenter
LLF:
Lactose
Non-Lactose Fermenter
4. Hektoen Enteric (HEA) Medium

Dark Green

Selective for: G (-) ENTERIC BACILLI

Inhibitors: Bile salts

citrate

Differential:
Indicators:
 RLF:

Lactose Fermenter ORANGE/YELLOW ORANGE

LLF:
Lactose
Non-Lactose Fermenter BLUE/GREEN
5. BISMUTH SULFITE (BSA) MEDIUM

Selective for:
Inhibitors:

Salmonella typhi colonies appear as Black colonies

with silver metallic sheen
6. BRILLIANT GREEN (BGA) MEDIUM

Selective for:
Inhibitors:

Salmonella spp. colonies appear as white colonies, snow

flake appearance and it is surrounded by Brilliant Red
agar
7. MANNITOL SALT (MSA) MEDIUM

Light/ salmon pink

Selective for: STAPHYLOCOCCUS ONY

7-10% salt concentration

Differential:
pH Indicators:

CHO incorporated: Mannitol

 Mannitol Fermenter

Mannitol
Non-Mannitol Fermenter
8. LOWENSTEIN JENSEN (LJ) MEDIUM

SELECTIVE FOR:
INHIBITORS:
SPUTUM SAMPLE NEEDS TO BE:

1. DECONGESTED/ DIGESTED
MUCIN/ MUCUS - N-ACETYL-L-CYSTEINE (NALC)

2. DECONTAMINATED
NaOH (Strong alkalinity)
• PHOTOCHROMOGEN- produces pigment in the
presence of light

• SCOTOCHROMOGEN- produce pigment in the absence

of light and intensifies pigment when exposed to light

• NON-CHROMOGEN- does not produce pigment on

presence/ absence of light
SELECTIVE MEDIUM FOR
NEISSERIA: BASICALLY
CAP WITH ANTIBIOTICS
INCORPORATED
A. THAYER MARTIN MEDIUM

GRAM POSITIVE INHIBITOR: Vancomycin

GRAM NEGATIVE INHIBITOR: Colistin
FUNGAL INHIBITOR: Nystatin
B. MODIFIED THAYER MARTIN (MTM) MEDIUM

GRAM POSITIVE INHIBITOR: Vancomycin

GRAM NEGATIVE INHIBITOR: Colistin
FUNGAL INHIBITOR: Nystatin
PROTEUS SPP. SWARMING INHIBITOR:
TRIMETHOPRIM
C. MARTIN LEWIS (MLM) MEDIUM

GRAM POSITIVE INHIBITOR: Vancomycin

GRAM NEGATIVE INHIBITOR: Colistin
FUNGAL INHIBITOR: ANISOMYCIN
PROTEUS SWARMING INHIBITOR: TRIMETHOPRIM
D. NEW YORK CITY (NYC) MEDIUM

GRAM POSITIVE INHIBITOR: Vancomycin

GRAM NEGATIVE INHIBITOR: Colistin
FUNGAL INHIBITOR: AMPOTHERICIN B
PROTEUS SWARMING INHIBITOR: TRIMETHOPRIM
F. CULTURE MEDIA FOR ANTIBIOTIC
SUSCEPTIBILITY/ SENSITIVITY TESTING

1. MOST BACTERIA: MUELLER HINTON AGAR

2. Haemophilus spp. : MHA + BLOOD
3. Mycobacterium spp. MIDDLEBROOK CM
G. CULTURE MEDIA FOR MOTILITY
H. ANAEROBIC CULTURE MEDIA
CYSTINE
REDUCING AGENTS:
THIOGLYCOLATE

EX. BACTEROIDES BILE ESCULIN MEDIUM

KANAMYCIN-VANCOMYCIN LAKED BLOOD AGAR
 PLATING MEDIA FOR
ROUTINE BACTERIOLOGY