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Bile Aesculin Agar, BEA Agar, is a selective and differential medium. This medium tests the ability of organisms to hydrolyze esculin in the presence of bile. It is commonly used to identify members of the genus group D streptococci and enterococci.
BEA medium contains bile to inhibit the growth of gram-positive bacteria as well as many gram-negative bacteria, and esculin which is the differential component.
In some preparations, the medium may contain sodium azide (increases the selectivity)
BEA agar
Suspend the components, dehydrated powder, in water (64g in 1000ml of purified/distilled water). Bring the medium to a boil with constant stirring for at least 1 minute.
Sterilize in autoclave at 121°C for 15 minutes, cool to 45-50°C.
BEA agar composition | |||
---|---|---|---|
Ingredients | Gram/liter | ||
Beef extract | 11g | ||
Enzymatic digestion of gelatin | 34,5 g/L | ||
Esculin | 1g | ||
manganese sulfate | 0.05g | ||
Ox bile | 2g | ||
Ferric Ammonium Citrate | 0,5 g/L | ||
Agar | 15,0 g/L |
◈ Note : the medium may contain sodium azide which increases selectivity, but sodium azide is extremely dangerous.
◈ Gelatin peptone and beef extract provide the essential elements needed for growth.
◈ Differentiation : If an organism can hydrolyze aesculin in the presence of bile, the product esculetin is formed. Esculetin reacts with ferric citrate (in the middle), forming a phenolic iron complex that transforms the middle from dark brown to black .
◈ Selectivity : Selectivity is obtained by the addition of bile (oxgall), which inhibits the growth of most Gram-positive cocci other than group D streptococci and enterococci.
◈ Examine after 18 to 24 hours for esculinase positive colonies. Wait up to 72 hours before reporting as negative.
Bacteria | Growth |
---|---|
Enterococcus | Good growth. Positive esculin hydrolysis resulting in darkening of the medium around the growth. |
Escherichia coli | Good growth. No darkening of media around colonies |
Streptococcus pyogenes | No growth |
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