TM 336 EMB Agar Data Sheet

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EMB AGAR or EOSIN METHYLENE BLUE AGAR is used for isolation and differentiation of gram negative enteric bacilli from clinical and non-clinical samples. Primarily this medium was used for the detection and confirmation of coliforms. Medium contains Lactose and Sucrose, due to these carbohydrates sources medium can be differential in primary culture: salmonellas and shigellas which are lactose-negative can be differentiated from other lactose - negative but sucrose - positive organisms such as Proteus sp.

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www.tmmedia.in PRODUCT DATA SHEET Manufacturer Address: Titan Biotech Limited A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Authorized Representative: MedNet GmbH Borkstrasse 10 48163 Munster Germany. Page 1 EMB AGAR TM 336 INTENDED USE For differential isolation of gram-negative enteric bacilli from clinical nonclinical samples COMPOSITION Ingredients Gms/Ltr Agar 13.500 Peptone 10.000 Lactose 5.000 Sucrose 5.000 Dipotassium phosphate 2.000 Eosin Y 0.400 Methylene blue 0.065 PRODUCT SUMMARY AND EXPLAINATION EMB AGAR or EOSIN METHYLENE BLUE AGAR is used for isolation and differentiation of gram negative enteric bacilli from clinical and non-clinical samples. Primarily this medium was used for the detection and confirmation of coliforms. Medium contains Lactose and Sucrose due to these carbohydrates sources medium can be differential in primary culture: salmonellas and shigellas which are lactose-negative can be differentiated from other lactose - negative but sucrose - positive organisms such as Proteus sp. PRINCIPLE Peptone is the nitrogen source in EMB Agar. Agar is a solidifying agent. Dipotassium phosphate is the buffer. Eosin Y and Methylene Blue are the indicators. Methylene blue is also a selective agent. The accompanying micro flora which hinders the isolation of medically important organisms is inhibited by the dyes of the medium especially gram -positives. The dyes in EMB agar Eosin Y and M ethylene blue are pH indicators and inhibitors of gram -positive bacteria and at an acid pH combine to form a green -metallic precipitate. Colonies of lactose fermenters are blue -black with or without a green metallic sheen. Sheen production appears to be sensitive to changes in pH and the lack of sheen production could be due to the alkalinity of mastitic milk interfering with the acidic requirement of EMB agar for production of the green metallic sheen. Colonies of non -lactose fermenting bacteria are colourless and translucent.

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www.tmmedia.in PRODUCT DATA SHEET Manufacturer Address: Titan Biotech Limited A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Authorized Representative: MedNet GmbH Borkstrasse 10 48163 Munster Germany. Page 2 INSTRUCTIONS FOR USE 1. Dissolve 36.0 gms in 1000ml of distilled water. 2. Gently heat to boiling with gentle swirling and dissolve the medium completely. 3. Sterilize by autoclaving at 15 psi 121º for 15 minutes. 4. Cool to 45-50 °C and after mixing dispense as desired. QUALITY CONTROL SPECIFICATIONS Appearance Dehydrated powder: Light pink to purple colour free flowing powder Appearance of the prepared medium: Reddish purple colour opalescent gel with greenish cast finely dispersed ppt. pH at 25°C: 7.2 ± 0.2 INTERPRETATION: Cultural characteristics observed after incubation at 35 ± 2°C for 18 – 24 hours. Microorganisms ATCC Inoculum CFU/ml Growth Appearance of colony Escherichia coli 25922 50-100 Good Black with green metallic sheen Enterococcus faecalis 29212 50-100 Partial inhibition ------------- Pseudomonas aeruginosa 27853 50-100 Fair - Good Colourless Staphylococcus aureus 13150  1000 Inhibited Colourless STORAGE STABILITY Dehydrated powder hygroscopic in nature store in a dry place in tightly-sealed containers below 25°C and protect from direct Sunlight. Under optimal conditions the medium has a shelf life of 4 years. When the container is opened for the first time note the time and date on the label space provided on the container. After the desired amount of medium has been taken out replace the cap tightly to protect from hydration. REFERENCES 1. A.P.H.A Examination of dairy products. 10 th Ed. APHA Inc. New York 1953. Society of American Bacteriologists. Manual of Microbiological Methods MacGraw -Hill New York 1957. 2. Holt -Harris J. E. and O. Teague. A new culture medium for the isolation of Bacillus typhosa

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www.tmmedia.in PRODUCT DATA SHEET Manufacturer Address: Titan Biotech Limited A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Authorized Representative: MedNet GmbH Borkstrasse 10 48163 Munster Germany. Page 3 from stools. J. Infect. Dis. 18 : 596. 1916. NOTE: Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

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