Français

Microbiological
technique  Cled Agar | Principle | Interpretation

☰ Content :

Ⅰ. Overview:
Ⅱ. Preparation
ⅠⅡ. Principle of Cled Agar
Ⅳ. Interpretation of Cled Agar
Ⅴ. Frequently asked questions

Ⅰ. Overview

Cled agar (Cystine Lactose

Electrolyte Deficient ) is
differential, non selective culture
medium used for the isolation,
enumeration and differentiation of
urinary microorganisms. It
promotes the growth of urinary
pathogens, but prevents excessive
swarming of Proteus species due
to its lack of electrolytes.

The medium allows quantitative Cled agar

determination of urinary
pathogens including Proteus when
calibrated loops are used for
inoculation.

Ⅱ. Preparation / composition

Suspend the components, dehydrated powder, in water (36.13 grams in 1000 ml of

purified/distilled water). The medium is boiled for a few seconds until the ingredients are
completely dissolved. Mix well and finally sterilize by autoclaving at 121°C for 15
minutes.

Cool to 50-55°C and dispense aseptically into sterile Petri dishes. Date the medium and
assign it a lot number.

Cled agar composition

Ingredients gram/liter Ingredients gram/liter

Balanced 4.0 g Tryptone 4.0 g

Peptone

Beef 3.0 g Lactose 10.0 g

extract

L-Cystine 0.128 g Bromothymol 0.02 g

CLED Agar (Dehydrated)
Blue

Agar 15.0 g Final pH 7.3 +/-

0.2

 Ⅲ. Principle of Cled Agar

◈ Originally described by G.H. Sandys in 1960, the medium contains cystine, lactose,
bromothymol blue and is electrolyte deficient.

Cystine is added for the benefit of organisms that have a specific need for cystine.
It promotes the growth of coliforms.
Lactose is included to provide an energy source for organisms capable of utilizing
it through a fermentation mechanism.
Bromothymol blue is the indicator used in the agar, it turns yellow in case of acid
production during lactose fermentation or turns dark blue in case of alkalinization.
Lactose-positive bacteria form yellow colonies. Bacteria that decarboxylate L-
cystine cause an alkaline reaction and form dark blue colonies
Electrolyte deficiency reduces the invasion of the environment by Proteus.

Ⅳ. Interpretation of Cled Agar

Bacteria Growth

Escherichia coli, Yellow colonies, opaque; yellow medium

Citrobacte

Enterobacter, Klebsiella Yellow to whitish-blue colonies, often mucoid; yellowish

medium

Proteus Translucent blue colonies; blue-green to blue medium

Pseudomonas Green colonies with typical matted surface and rough

periphery; blue medium

Streptococci Small, opaque, pale yellow colonies

Staphylococci Very small, opaque, yellow colonies

Enterococci Small yellow colonies, about 0.5 mm in diameter; yellow

medium


 Ⅴ. Frequently asked questions

Q : Why CLED agar is electrolyte deficient?

A : CLED agar is electrolyte deficient to prevent excessive swarming of Proteus species.
Crisley showed that a strain of Proteus vulgaris grown in the presence of sodium chloride
utilized more glucose and grew to a greater extent than when grown in the absence of
sodium chloride. Naylor showed that this growth enhancement, in the presence of NaCl,
is a factor in causing Proteus to swarm.

Q : Is Cled selective?
A : CLED agar is a non-selective medium, It allows the growth of a wide variety of non-
fastidious germs

Q : Does Salmonella grow on CLED?

A : Yes, salmonella can grow on CLED, salmonella doesn't utilize lactose (blue-green
colonies on the surface of CLED agar).

Q : Why does E coli grow as yellow colonies on CLED?

A : Organisms capable of fermenting lactose, like E coli, will lower the pH and change the
color of the medium to yellow (colonies appear yellowish because the agar will turn
yellowish)

Reference

1. Sandys, G. H. - A New Method of Preventing Swarming of Proteus sp. with a Description of a New Medium suitable for Use in
Routine Laboratory Practice
2. NEOGEN - Cystine Lactose Electrolyte Deficient (CLED) Agar
3. universe84a - CLED Agar
4. P. G. D. NAYLOR - Effect of Sodium Chloride, Dulcitol and Glucose on the Growth of Proteus in Shaken and Static Broth Cultures
5. Crisley - EFFECT OF SODIUM CHLORIDE ON GROWTH, GLUCOSE UTILIZATION, AND ACID PRODUCTION IN PROTEUS VULGARIS

6. BD- CLED Agar

7. sigmaaldrich- CLED Agar
8. ACUMEDIA- CLED Agar


report this ad

Tags About us Popular

Antibiotic susceptibility testing Stains Contact us About us Gram stain Coagulase test Microbiological
Microbiological Techniques diagnosis
Laboratory Equipment

Powered By : MWL @2022