Ingredient g/L
Casein enzymic hydrolysate 5.00
Proteose peptone 5.00
Meat extract B# 5.00
Yeast extract 5.00
Sodium chloride 20.00
Monopotassium dihydrogen phosphate 1.35
Disodium hydrogen phosphate 12.00
Esculin 1.00
Nalidixic acid 0.02
#- Equivalent to Beef extract
Final pH (at 25°C) : 7.4 + 0.2
Direction
Suspend 54.37 gms.in 1000ml. distilled water.Heat to boiling to dissolve the medium completely.Sterilize by autoclaving at 15 lbspressure (121°C) for 15 minutes. Cool to 50°C and aseptically add filter sterilized solution of acriflavin hydrochloride to a final Concentration of 12 mg per litre of medium or 25 mg per litre as per the requirement.
Principle
Casein enzymic hydrolysate, proteose peptone, beef extract and yeast extract provide necessary nutrients while esculin offers differential properties to the medium. Nalidixic acid and acriflavin hydrochloride together with higher concentration of phosphate render the medium selective for Listeria, as the gram – negative and gram – positive organisms are inhibited by Nalidixic acid and acriflavin hydrochloride respectively.
QC Tests
(I) Dehydrated Medium
Colour : Cream to yellow
Appearance : Homogeneous Free Flowing powder
(II) Rehydrated medium
pH (post autoclaving/heating) : 7.4 ± 0.2
Colour (post autoclaving/heating) : Medium amber
Clarity (post autoclaving/heating) : Slightly opalescent
(III) Q.C. Test Microbiological
Cultural characteristics observed after 24- 48 hours at 35-37°C.
MICROORGANISM (ATCC ) GROWTH
Listeria monocytogenes (19118 ) Good – luxuriant
Escherichia coli (25922) None – poor
Staphylococcus aureus (25923) None – poor
Use
For selective isolation and cultivation of Listeria monocytogenes from clinical specimens.
Storage
Dehydrated medium andprepared medium– Between 2 to 8°C.