Deskripsi

Himedia M021-500G Triple Sugar Iron Agar

Himedia M021-500G Triple Sugar Iron Agar

Composition**

Ingredients Gms / Litre
Peptone 10.000
Tryptone 10.000
Yeast extract 3.000
HM Peptone B# 3.000
Lactose 10.000
Sucrose 10.000
Dextrose (Glucose) 1.000
Sodium chloride 5.000
Ferrous sulphate 0.200
Sodium thiosulphate 0.300
Phenol red 0.024
Agar 12.000
Final pH ( at 25°C) 7.4±0.2
**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 64.52 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Mix well and
distribute into test tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Allow the medium to set in
sloped form with a butt about 1 inch long.
Note: For better results, the medium can be sterilized by autoclaving at 10 lbs pressure (115°C) for 15 minutes.

Principle And Interpretation

Triple Sugar Iron Agar was originally proposed by Sulkin and Willett (8) and modified by Hajna (3)
for identifying Enterobacteriaceae . This medium complies with the recommendation of APHA, for the examination of
meat and food products (7), for the examination of milk and dairy products (9) and for microbial limit test for confirming
the presence of Salmonellae (1,2) and in the identification of gram-negative bacilli (1, 6).

Tryptone, peptone, yeast extract and HM peptone B provide nitrogenous compounds, sulphur, trace elements and
vitamin B complex etc. Sodium chloride maintains osmotic equilibrium. Lactose, sucrose and dextrose are the
fermentable carbohydrates. Sodium thiosulphate and ferrous ions make H2S indicator system. Phenol red is the pH
indicator. Organisms that ferment glucose produce a variety of acids, turning the colour of the medium from red to yellow.
More amount of acids are liberated in butt (fermentation) than in the slant (respiration).

Growing bacteria also form alkaline
products from the oxidative decarboxylation of peptone and these alkaline products neutralize the large amounts of acid
present in the butt. Thus the appearance of an alkaline (red) slant and an acid (yellow) butt after incubation indicates that
the organism is a glucose fermenter but is unable to ferment lactose and/or sucrose. Bacteria that ferment lactose or
sucrose (or both), in addition to glucose, produce large amounts of acid enables no reversion of pH in that region and thus
bacteria exhibit an acid slant and acid butt.

Gas production (CO2)

is detected by the presence of cracks or bubbles in the medium, when the accumulated gas escapes. Thiosulphate is reduced to hydrogen sulphide by several species of bacteria and
H2S combines with ferric ions of ferric salts to produce the insoluble black precipitate of ferrous sulphide. Reduction of
thiosulphate proceeds only in an acid environment and blackening usually occurs in the butt of the tube. Triple Sugar Iron
Agar should be used in parallel with Urea Agar/Broth (M112/M111) to distinguish between Salmonella and Proteus
species. The reactions can be summarized as follows:
I

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