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Triple Sugar Iron Agar Technical Datasheet: TM Media

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

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Triple Sugar Iron Agar Technical Datasheet: TM Media

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  1.   TM Media   904, 9th Floor, Bigjos Tower, Netaji Subhash Place, Delhi, 110034  Triple Sugar Iron Agar  INTENDED USE  Triple Sugar Iron Agar​ For confirmation of gram negative enteric bacilli on basis of  dextrose, lactose and sucrose fermentation and H2S production  COMPOSITION    Ingredients  Gms/Ltr.  Agar  12.000  Lactose  10.000  Peptic digest of animal tissue  10.000  Casein enzymatic hydrolysate  10.000  Sucrose  10.000  Sodium chloride  5.000  Yeast extract   3.000  Beef extract   3.000  Dextrose monohydrate  1.000  Sodium thiosulphate   0.300  Ferrous sulphate   0.200  Phenol red   0.024     

  2. 2   PRODUCT SUMMARY AND EXPLANATION  In 1911, Russell described a medium containing dextrose and lactose for differentiation             of enteric gram-negative bacilli. Krumwiede and                         Kohn modified Russell's double sugar medium             by adding sucrose which allowed for detection               of sucrose-fermenting gram-negative bacilli. In           1940, Sulkin and Willet further modified the               medium by adding ferrous sulfate, an indicator               of H2S production. Hajna developed the recent               formulation for Triple Sugar Iron Agar by adding                 sucrose to Kligler Iron Agar.  Triple Sugar Iron Agar is recommended for       differentiation of enteric, gram-negative bacilli                   from dairy samples and food products. This               medium   also   complies   with   the   recommendation of Indian Pharmacopoeia for the identification of Gram-negative                   bacilli.    PRINCIPLE  Peptone, yeast extract and beef extract provide nitrogenous compounds, sulphur, trace  elements and vitamin B complex etc. Sodium chloride maintains osmotic equilibrium.  Lactose, sucrose and dextrose monohydrate are the fermentable carbohydrates.  Sodium thiosulphate and ferric or ferrous ions make H2S indicator system. Sodium  thiosulphate is also an inactivator of halogen and can minimize its toxicity in the testing  sample, if any during microbial limit tests. Phenol red is the pH indicator. The dextrose  concentration in TSI Agar is one-tenth the concentration of lactose and sucrose which  serves to distinguish dextrose-only fermenting organisms from those which ferment   

  3. 3   lactose and/or sucrose. The small amount of acid produced in the slant during  dextrose fermentation oxidizes rapidly, causing the slant to revert to alkaline (red) while  the yellow acid reaction is maintained in the butt due to the absence of oxygen. If the  lactose or sucrose is also fermented, sufficient acid is produced to retain the yellow  color on the slant and in the butt. Ferric ammonium citrate and sodium thiosulfate are  indicators of H2S production which form a black precipitate (ferrous sulfate) in the butt  of the tube. Gas production is indicated by bubbles, splitting of the agar, or  displacement of the agar in the tube.   INSTRUCTION FOR USE  1. Dissolve 64.52 grams in 1000 ml distilled water.   2. Gently heat to boiling to dissolve the medium completely.   3. Mix well and distribute into test tubes.   4. Sterilize by autoclaving at 15 psi (121°C) for 15 minutes.   5. Allow the medium to set in sloped form with a butt about 2.5cm long.   QUALITY CONTROL SPECIFICATIONS  Appearance of Powder:​ Light yellow to pink colour, homogeneous free flowing powder  Appearance of prepared medium:​ Pinkish red colour, clear to slightly opalescent gel  pH (at 25°C) : 7.4 ± 0.2  INTERPRETATION:    Culture characteristics observed after incubation period of 18 - 24 hours at 35 ± 2°C    Microorganisms  ATCC   Inoculum  (CFU)  Growth  Slant  Butt/Gas   H2S  Salmonella  typhimurium   14028  50-100  Luxuriant  Alkaline,  red   Acidic,  yellow/+ ve  +ve   

  4. 4   Escherichia coli  25922  50-100  Luxuriant  Acidic,  yellow   Acidic,  yellow/+ ve  -ve  Shigella flexneri  12022  50-100  Luxuriant  Alkaline,  red  Acidic,  yellow/- ve  -ve  Salmonella typhi  6539  50-100  Luxuriant  Alkaline,  red  Acidic,  yellow/- ve  +ve  Enterobacter  aerogenes  13048  50-100  Luxuriant  Acidic,  yellow   Acidic,  yellow/+ ve  -ve    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.     

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