Laboratorios conda, s


CAT Nº: 2050
For the isolation and differentiation of Enterococcus faecalis and E. faecium
Final pH 7.1 ± 0.2 at 25ºC

Suspend 71.48 grams of the medium in one liter of distil ed water. Mix wel and dissolve by heating with frequent
agitation. Boil for one minute until complete dissolution. Sterilize in autoclave at 121ºC for 15 minutes. Cool to 50ºC, mix
well and dispense into plates. For a more selective medium, prepare a solution of 0.24 grams of nalidixic acid in 5 ml of
sterile distil ed water with a few drops of sodium hydroxide 0.1N (for a better dissolution), and aseptical y add to one
liter of medium. The prepared medium should be stored at 8-15°C. The color is amber, slightly opalescent.
The dehydrated medium should be homogeneous, free-flowing and beige in color. If there are any physical changes,
discard the medium.
Caution: This medium contains Sodium azide and Cycloheximide and it is very toxic if swallowed, inhaled or comes into contact with
skin. Wear gloves and eye face protection.
m-EI CHROMOGENIC AGAR BASE, MODIFIED is recommended for the isolation and differentiation of Enterococcus
faecium and Enterococcus faecalis.
This medium is a modification of the mEI cromogenic Agar base, where another chromogenic substrate is added. This
addition al ows the differentiation of Enterococcus faecium and Enterococcus faecalis.
Peptone provides nitrogen, vitamins, minerals and amino acids essential for growth. Yeast extract provides trace
elements, vitamins and amino acids. Esculin is hydrolyzed by enterococci to form esculetin and dextrose. Cycloheximide
inhibits most fungi, and the sodium azide inhibits Gram negative bacteria. Chromogenic Mix is added to differentiate
Enterococcus faecium from Enterococcus faecalis. Bacteriological agar is the solidifying agent.
Inoculate and incubate to 41±0.5 °C and observe alter 18-24 hours. Enterococcus faecium will grow as blue-greenish,
Enterococcus faecalis wil grow as intense blue colonies.
The fol owing results were obtained in the performance of the medium from type cultures, with the nalidixic acid added,
after incubation at a temperature of 41 ± 0.5°C and observed alter 24-48 hours.
Colony Color

Levin, Fischer and Cabelli. 1975. Appl. Microbiol. 30.66.
U.S. Environmental Protection Agency. 2002. Method 1600: Enterococci in water by membrane filtration using membrane enterococcus
indoxyl –D- glucoside agar (mEl]. Publication EPA-821- R-02-022. USEPA Office of Water, Office of Science and Technology, USEPA,
Washington, DC.
Once opened keep powdered medium closed to avoid hydration.


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