Fluorocult ECD Agar EMD Chemicals Inc.

Fluorocult^® ECD Agar

EMD Cat. No. 1.04038.0500

500 g

The medium complies with the German-DIN-Norm 10110 for the examination of meat, with the regulations acc. to § 35 LMBG (06.00/36) for the examination of food and with ISO Standard 6391 (1996) for the enumeration of E. coli in meat and meat products.

Experimental Procedure and Evaluation	Quality Control
Typical Composition (g/liter)	Literature

Mode of Action
The bile salt mixture of this E. coli Direct Agar largely inhibits the accompanying flora not usually found in the intestines. Using fluorescence under UV light and a positive indole reaction, E. coli colonies can be identified among the grown colonies.

Typical Composition (g/liter)
Peptone from casein 20.0; lactose 5.0; sodium chloride 5.0; bile salt mixture 1.5; di-potassium hydrogen phosphate 4.0; potassium dihydrogen phosphate 1.5; agar-agar 15.0; tryptophan 1.0; 4-methylumbelliferyl-β-D-glucuronide 0.07.

Preparation
Suspend 53.1 g in 1 liter of purified water. Autoclave (15 min at 121°C). Pour plates.
pH: 7.0 ± 0.2 at 25°C.
The prepared broth is clear and yellowish brown.

Experimental Procedure and Evaluation
The culture medium is inoculated in the usual way by streaking on the surface and uncubated for 18-24 hours at 44 °C aerobically.

Fluorescence is noted with a UV lamp: light blue fluorescing colonies identify E. coli.

For confirmation cover the colonies with 10-20µl KOVACS indole reagent, e.g. using Bactident^® Indole. A reddening after 2-10 seconds shows indole formation.

Quality control

Test strains	Growth / Recovery rate %	MUG	Indole
Escherichia coli ATCC 8739	>= 70	+	+
Escherichia coli ATCC 25922	>= 70	+	+
Enterobacter aerogenes ATCC 13048	good / very good	-	-
Klebsiella pneumoniae ATCC 13883	good / very good	-
Citrobacter freundii ATCC 8090	good / very good	-
Proteus mirabilis ATCC 14153	good / very good	-
Pseudomonas aeruginosa ATCC 27853	good / very good
Clostridium perfringens ATCC 10⁵43	none / poor (anaerobic)

Additives

EMD Cat. No.	Product	Pack Size
1.09293.0100	KOVÁCS Indole Reagent	100 ml
1.11350.0001	Bactident^® Indole (dropper bottle)	1 x 30 ml
1.13203.0001	UV Lamp (366 nm)	1 ea

Literature

Bundesgesundheitsamt: Amtliche Sammlung von Untersuchungsverfahren nach § 35 LMBG. - Beuth Verlag Berlin, Köln
DIN Deutsches Institut für Normung e.V.: Mikrobiologische Fleischuntersuchung. Bestimmung der Escherichia coli. Fluoreszenzoptisches Koloniezählverfahren unter Verwendung von Membranfiltern/Spatelverfahren (Referenzverfahren). DIN 10110.
Draft International Standard ISO/DIS 6391: Meat and meat products - Enumeration of Escherichia coli-colony-count technique at 44 °C using membranes (1996).