Exercise 5-21 Micro Lab: Triple Sugar Iron Agar / Kligler Iron Agar

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Triple Sugar Iron Agar

a rich medium designed to differentiate bacteria on the basis of glucose fermentation, lactose fermentation, sucrose fermentation, and sulfur reduction


What does triple sugar iron agar contain?

Three carbohydrates,
animal proteins as sources of carbon and nitrogen,
ferrous sulfate and sodium thiosulfate as sources of oxidized sulfur


______ is the pH indicator and _____ is the hydrogen sulfide indicator

phenol red
ferrous sulfate


How is triple sugar iron agar prepared?

in a shallow agar slant with a deep butt. it is inoculated by a stab in the agar butt followed by a fishtail streak of the slant


Incubation period for triple sugar iron agar

18-24 hrs for carbohydrate fermentation
48 hrs for hydrogen sulfide reactions



TSIA is inoculated with a glucose only fermenter. As the glucose diminishes, the organisms located in the aerobic region (slant) will break down amino acids, producing NH3 and raising the pH. This process takes 18-24 hours and only occurs in the slant because of the anaerobic conditions in the butt.


TSIA slant with a red slant and yellow butt indicates what?

that the organism ferments glucose but not lactose


Organisms that ferment both glucose and lactose/sucrose do what to the medium?

turn it yellow throughout


_______ produced by fermentation of any of the carbs will appear as fissures in the medium or will lift the agar off the bottom of the tube



Why is hydrogen sulfide produced?

by the reduction of thiosulfate in the medium or by the breakdown of cysteine in the peptone


black precipitate in the medium is an indication of what?

sulfur reduction and fermentation


red slant indicates what?
yellow slant indicates what?

red - glucose fermentation
yellow - glucose and lactose fermentation

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