Carbs excersise 21 triple sugar Iron Agar test

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What is TSI test used for

Identify enteric bacteria and differentiate them from other gram neg rods



Rich medium differentiates glucose fermentation, lactose fermentation, sucrose fermentation and sulfur reduction


What does TSIA include in addition to 3 carbs

Beef extract
Yeast extract
Sodium thiosulfate


What is peptone used for

Carbon and nitrogen source


What is sodium thiosulfate used for

Source of reducible sulfur


What is pH indicator in TSIA

Phenol red


What is the hydrogen sulfide indicator in TSIA

The iron in ferrous sulfate


How is TSIA medium prepd

Agar slant with deep butt provides aerobic and anaerobic growth enviornments


How is TSIA slant inoculated

Stab in agar butt followed by fishtail streak of slant


How long is incubation

18-24 hrs for carb ferm
48 hrs for hydrogen sulfide



Those organisms capable of breaking glycosidic bonds in carbs


TSI yellow slant/yellow butt

Glucose and lactose and or sucrose fermentation with acid accumulation in slant and butt


What happens when TSI inoculated with glucose only fermenter

Acid products lower pH turn everything yellow I. A few hrs but glucose in short supply(.1) exhausted in 12 hrs


What happens as glucose used up

Reversion, takes 18-24 hrs Organisms located in slant (aerobic region) start to break down available amino acids producing NH3 and raising pH


Where does reversion occur

Only in slant bc of anaerobic conditio s in butt


What does a TSI slant with a red slant and yellow butt after 24 hrs incubation indicate

That organism ferments glucose but not lactose


What color is slant if organisms ferment glucose and lactose

Yellow throughout but bc lactose and sucrose 10 higher conc than glucose, more acid produced and slant and butt stay yellow after 24 hrs


TSI test result red slant/ yellow butt

Glucose fermentation with acid production
Proteins catabolized aerobically (in slant) with alkaline products (reversion)


TSI test result red slant/red butt

No fermentation
Peptone catabolized aerobically and anaerobically with alkaline oproducts
Not from enterobacteriacae


TSI test result red slant/no change in butt

Organism is growing slowly or not at all
Not from enterobacteriacae


Black precipitate in agar

Sulfur reduction (an acid condition, from fermentation of glucose or lactose and or sucrose, exists in the butt even if the yellow color is obscured by black precipitate


Cracks in or lifting of agar

Gas production


What could happen if read results too early

Early- could show yellow throughout leading you to think org is lactose or sucrose fermenter when it simply may not have gotten rid of glucose yet


Reading after lactose and sucrose depleted

Would show yellow butt red slant so think org is glucose only fermenter


What is purpose of thiosulfate in TSI agar

When bacteria use sodium thiosulfate they can reduce it to sulfite using enzyme thiosulfate reductase and release hydrogen sulfide gas


What is meant by term saccharolytic bacteria. What reaction would it give in TSI tube

Uses all three sugars, glucose, lactose, sucrose
Slant would be yellow bc acid formation from breaking glycosidic binds

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