The Triple Sugar Iron (TSI) Test - Procedure, Uses

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Triple Sugar Iron Agar is used (TSI Agar)for confirmation of gram negative enteric bacilli on basis of dextrose, lactose and sucrose fermentation and H2S production or confirmation of gram negative enteric bacilli on basis of dextrose, lactose and sucrose fermentation and H2S production.

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www.titanmedia.in Page 1 TRIPLE SUGAR IRON AGAR AS PER BIS TM 1874 PRODUCT DATA SHEET For confirmation of gram negative enteric bacilli on basis of dextrose lactose and sucrose fermentation and H 2S production. Composition Ingredients Gms/Ltr Peptone 20.000 Yeast extract 3.000 Meat extract 3.000 Lactose 10.000 Sucrose 10.000 Dextrose 1.000 Sodium chloride 5.000 Ferrous sulphate heptahydrate 0.200 Sodium thiosulphate pentahydrate 0.300 Phenol red 0.024 Agar 12.000 Dehydrated powder hygroscopic in nature store in a dry place in tightly-sealed containers below 25°C and protect from direct Sunlight. Instructions for Use Suspend 64.32 grams in 1000 ml distilled water. Gently heat to boiling with gently swirling to dissolve the medium completely. Sterilize by autoclaving at 15 psi pressure 121°C for 15 mins. Allow the medium to cool at room temperature for preparing the slants. Appearance: Pinkish - red colour clear to slightly opalescent gel in slant position in screw cap tubes pH at 25°C : 7.4 ± 0.2. Principle Triple Sugar Iron Agar was originally proposed by Sulkin and Willet and modified by Hajna for identifying Enterobacteriaceae. ISO Committee and BIS has recommended a slight modification for the identification of Salmonellae. BIS has recommended the medium for detection of Escherichia coli and Vibrios. Triple Sugar Iron Agar is recommended for confirmation of gram negative bacilli on basis of dextrose lactose and sucrose fermentation and H 2S production. The ingredients included in the medium such as peptone yeast extract meat extract provide the nitrogen carbon and vitamins required for organism growth. Triple sugar Iron Agar Consists of three carbohydrates-dextrose Lactose and Sucrose. When the carbohydrate are fermented acid production is detected by the phenol red pH indicator. Sodium thiosulphate is reduced to hydrogen sulphide and hydrogen sulphide reacts with an iron salt yielding the typical black iron sulphide. Ferric ammonium

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www.titanmedia.in Page 2 PRODUCT DATA SHEET citrate is the hydrogen sulphide H 2S indicator. Sodium chloride maintains the osmotic balance of the medium. Agar is used as a solidifying agent. Interpretation Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours Test Strains ATCC Inoculum cfu/ml Growth Butt/Slants/Gas/H 2S Salmonella Paratyphi A 9150 10 3 Luxuriant Yellow /Red/+/- Salmonella Typhi 6539 10 3 Luxuriant Yellow /Red/-/+ Salmonella Typhimurium 14028 10 3 Luxuriant Yellow/Red/+/+ Vibrio cholerae 15748 10 3 Luxuriant Yellow/Red/-/- Reference 1. Sulkin E.S. and Willett J.C. 1940 J. Lab. Clin. Med. 2. Hajna A.A. 1945 J. Bacteriol 49:516. 3. Finegold and Baron 1986 Bailey and Scotts Diagnostic Microbiology 7th ed. The C.V. Mosby Co. St. Louis. 4. Greenberg A. E. Trussell R. R. and Clesceri L. S. Eds. 1985 Standard Methods for the Examination of Water and Wastewater 16th ed. APHA Washington D.C. 5. MacFaddin J. 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria Vol. 1 Williams and Wilkins Baltimore.

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