TSI test

TSI test

1. CHEMICAL TEST
TSI TEST: ( Triple Iron Agar Test )
TSI contains there sugar ;
(1) Glucose (2) Lactose (3) Sucrose
REQURIREMWENTS:
A. Graduated Cylinder .
B. Take Flask.
C. Test Tubes.
D. Cotton.
E. Aluminum foil sheet.
F. Electric Balance.
G. Digital balance .
H. Autoclave.
I. Straight wire loop.
CHEMICAL:
TSI Media , and Distilled Water
CALCULATON:
65 g = 1 litter (1000 ml)
X = 50 ml
Cross multiplication
�� ?????? × �� ��÷���� �� = 3.25 g
?????? ×���� �� ÷���� �� = 50 ml

PROCEDURE:
1. Take 50 ml distilled water with the help of Graduated Cylinder in the Flask.

2. 2. Weight the amount of Media for in gram on digital balance and pour it in the flask.
3. Shake the flask to make a proper solution of TSI agar.
4. After preparing the solution, pouring the Test tubes from the solution .
5. Keep the Test Tubes in the autoclave for sterilization and for more dissolving of the solution at
121 °C (249°F) for 15 minutes at 50 psi .
6. After autoclavation , take out the test tubes from autoclave and placed them in slightly tilted
position for the formation of slants and solidification of the media in the tubes in slant position.
7. After slant formation , perform inoculation ( stabling) in the tubes and incubated the slants for
24 hours at 37 °C .
8. After 24 hours observe the test tubes , the colors and composition of the media in the tubes will
be changed. Due to gas ( Hydrogen Sulphide ) production and also producing of cracks in the
media inside the tubes. Cracks produced due to Hydrogen Sulphide gas inside the media .
Yellow color of the media in the tubes determined by the fermentation of glucose or sucrose.
9. Different bacterial species changed the color of butt and slop.
o Sometime butt and slop both are yellow
o Sometime butt and slop both are black .
o Sometime butt is black and slop is yellow.
o Sometime butt is yellow and slop is blacked.

Prepared By Amjad Khan Afridi
Date : 16 / 12 / 2015