E.M.B. (EOSIN METHYLENE BLUE) AGAR

Technical Data #1268a / 2013.05.01

E.M.B. Agar is a solid medium for isolation and identification of gram-negative enteric bacteria in clinical and non clinical specimens. Addition of sucrose differentiate sucrose fermenters from sucrose non fermenters. Eosin Y and Methylene Blue dyes inhibit a large quantity of gram-positive and stain colonies of lactose and sucrose fermenters. Colonies with metallic sheen confirm Escherichia coli growth. Fecal streptococci will grow on this medium and form pinpoint colonies.

FORMULA
in grams per liter purified filtered water

Gelatin Peptone	10,0
Lactose	5,0
Sucrose	5,0
Dipotassium Phosphate	2,0
Eosin Y	0,4
Methylene Blue	0,065
Agar	13,5

pH 7,2 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store prepared media at 2-8º C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25º C.

SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.

DIRECTIONS
Suspend 36 g of dehydrated media in 1000 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C. Mix gently and dispense into sterile Petri dishes.

PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

1. Directly, inoculate plate as soon as possible with specimen and streak for isolation.
2. Incubate at 35º C for 24 hours.

QUALITY CONTROL
Results after 24 hrs at 35ºC

Organisms	ATCC	Growth	Colour
Escherichia coli	25922	+	black/metallic sheen
Salmonella typhimurium	14028	+	colourless
Pseudomonas aeruginosa	27853	+	colourless
Enterobacter aerogenes	13048	+	brownish
Streptococcus faecalis	29212	- or partial

LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.

REFERENCES

1. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American society for Microbiology.
2. Mac Faddin, Jean F., 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol.1. Baltimore, MD.: Williams & Wilkins.

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-1606
Prepared media
Plates, 100x15 mm	10/pkg	1268
Mono-Plates, 100x15 mm	10/pkg	1268M