Deoxycholate Citrate Agar Medium, Granulated

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GM065S
Recommended for isolation of Shigella species from food samples. It is recommended by BIS committee under the specifications IS:5887(Part I)-1999.


Intended Use

Desoxycholate citarte agar medium, granulated is recommended for Isolation of Shigella species from food samples in with IS 5887(Part 7):1999.

Composition

Ingredients Gms / Litre
Meat extract 4.550
Proteose peptone 4.550
Lactose 9.090
Neutral Red 0.023
Sodium citrate 7.720
Sodium thiosulphate 7.720
Ferric Ammonium citrate 0.900
Sodium Desoxycholate 0.450
Agar 20.450
Final pH (at 25°C) 7.3±0.2

Formula adjusted, standardized to suit performance parameters

Directions

Suspend 55.45 grams of medium in 1000 ml of distilled water. Heat to boiling to dissolve the medium completely. DO NOT AUTOCLAVE. Avoid excessive heating as it is detrimental to the medium. Cool to 45-50°C and pour into sterile Petri plates. Dry the surface medium before incubation.

Principle And Interpretation

The described selective medium in accordance with IS 5887 (1999) Part 7(1) and is recommended for plating the growth obtained from Selenite F broth of Tetrathionate broth.

Sodium deoxycholate is inhibitory to gram positive bacteria. Most intestinal flora gets inhibited due to citrate in the medium. Coliforms produce pink colored colonies whereas non lactose fomenters appear as colourless colonies. The organisms known to reduce ferric ammonium citrate to iron sulphide are indicated by blackening of the central position of colony.

Meat Extract and Proteose peptone serves as a source of carbon and nitrogen. Lactose is the fermentable carbohydrate and neutral red acts as pH indicator helping in differentiaion of enteric bacilli as lactose ferments produce red colonies while non-lactose fermentors produce colourless colonies.

In general all foods which have received dry heat treatment, namely desiccation, dehydration or powdering shall be enriched in a non-inhibitory medium such as nutrient broth. Raw foods and finished products which are suspected to be grossly contaminated after processing does not require pre-enrichment. Approximately 200 ml Selenite F broth per approximately 25 g of one portion of sample and 200 ml of tetrathionate broth per approximately 5 g of another portion of the sample is blended in sterile blender jar for 2 minutes or macerated with sterile sand in a sterile mortar. Incubation is carried out at 37°C for 24 hours. Raw meat samples should be incubated at 43°C, rather than 37°C. Further, it can be plated on Deoxycholate Agar (GM065S/ M065S)

Quality Control

Appearance
Light yellow to pinkish beige coloured granular medium

Gelling
Firm, comparable with 2.045% Agar gel.

Colour and Clarity of prepared medium
Reddish orange coloured, clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 5.55% w/v aqueous solution at 25°C. pH: 7.3±0.2

Cultural Response

Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.

Organism Inoculum (CFU) Growth Recovery Colour of Colony H2S
Enterococcus faecalis ATCC 29212 50-100 inhibited 0%
Escherichia coli ATCC 25922 50-100 poor 20-30% pink with bile precipitate negative reaction
Salmonella Enteritidis ATCC 13076 50-100 good-luxuriant >=50% colourless positive reaction, black centered colonies
Salmonella Typhimurium ATCC 14028 50-100 good-luxuriant >=50% colourless positive reaction, black centered colonies
Shigella flexneri ATCC 12022 50-100 good 40-50% colourless
Shigella sonneii ATCC 29930 50-100 good 40-50% colourless negative reaction
Escherichia coli ATCC 8739 50-100 poor 20-30% pink with bile precipitate negative reaction
Escherichia coli NCTC 9002 50-100 poor 20-30% pink with bile precipitate negative reaction

Storage and Shelf Life

Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.

Reference

  1. Bureau of Indian standard, IS 5887 (Part7) 1999. Methods for detection of bacteria responsible for food poisoning.
More Information
Product Name Deoxycholate Citrate Agar Medium, Granulated
SKU GM065S
Product Type Granulated
Physical Form Granular
Origin Animal
Packaging type HDPE
References 1. Leifson, 1935, J. Path. Bact., 40:581.
2.Speck M. (Ed.), 1984, Compendium of Methods for the Microbiological Examination of Foods, 2nd ed., APHA,Washington, D.C.
3.Frieker C.R., 1987, J. Appl. Bact., 63:99.
4.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
5.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1.
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