Phenol red: Indicator of acidification (It is yellow in acidic condition and red under alkaline conditions).
Addition of sucrose in TSI Agar  permits  earlier detection of coliform bacteria that ferment sucrose more rapidly than lactose. If lactose is not fermented but the small amount of glucose is, the oxygen deficient butt will be yellow (remember that butt comparatively have more glucose compared to slant i.e. Please repeat the procedure (following all the steps and precautions as mentioned) with a single isolated colony; and observe the result within 18-24 hours.
The rationale is, MTB concentrate in the sputum at night and sensitivity increases if early morning sample is taken. Enter your email address to subscribe to this blog and receive notifications of new posts by email.

Adding sucrose also aids the identification of certain gram-negative bacteria that could ferment sucrose but not lactose. Other basic understanding is TSI Tube contains butt (poorly oxygenated area on the bottom) slant (angled well oxygenated area on the top). The slant can become a deeper red-purple (more alkaline) as a result of production of ammonia from the oxidative deamination of amino acids (remember peoptone is a major constitutents of TSI Agar) . When the H2S is produced in the slant, it get escaped but in the slant, it get trapped and react with Iron, resulting the black pigmentation..
If organism is fermenter, it will ferment at the butt also, if organism is non-fermenter, it will oxidize the peoptone and alkali condition will come. Based on the information you have provided here, the isolate can be one of them; Escherichia coli, Klebsiella spp (pneumoniae or oxytoca), Enterobacter spp or Citrobacter spp. In our settings; we do not perform Cholera Red Reaction as a part of routine test to identify Vibrio cholerae.

Blogging is my passion, I am working as a lecturer and Microbiologist at Department of Microbiology and Immunology, Patan Academy of Health Sciences, Lalitpur. We perform simple test from Hanging drop (for rice watery stool with mucus flecks), followed by plating in TCBS agar.
And the Kligler Iron Agar was completely yellow with one or two small bubbles at the bottom.
You need other information to identify the isolate such as production of Hydrogen sulphide gas, indole production, citrate utilization, urease production, motility etc.

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