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4.7: Triple Sugar Iron Medium

  • Page ID
    151848
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    Triple Sugar Iron is a slant medium with two growth environments: aerobic (on the slant) and anaerobic (in the “butt”). The medium contains three sugars in varying concentrations and a pH indicator that turns yellow at pH measurements below 6.8, and a deeper red at pH measurements above 8.2. Bacteria that ferment typically produce one or more types of acid as a byproduct, therefore, fermentation (both aerobic on the slant and anaerobic in the butt) is noted as a change in the color of the media. The medium also identifies strict aerobes that only grow on the slant surface, and also bacteria that produce H2S, either as a way to produce ATP anaerobically using sulfur or sulfate as a final electron acceptor, or as a result of the breakdown of proteins that contain high numbers of sulfur-containing amino acids (cysteine or methionine).

    The results of this test are reported as appearance of the slant/appearance of the butt, using A to indicate acid reaction (yellow color), K to indicate an alkaline reaction, and NC to indicate no change in the medium. H2S (detected as a blackening in the media) and the production of gas (CO2) as a byproduct of fermentation are also reported if observed (see Figure 4).

    As an example, and for practice, the interpretation and outcomes for the 4 TSI tests shown are provided in the table below. Note that many other possible reactions may also occur so proper interpretation of this test is important.

    image4-2.jpeg
    Table 1. TSI reactions shown in the cultures in Figure 4, from left to right.
    Outcome Interpretation
    Uninoculated control For color comparison with inoculated samples
    K/NC Aerobic respiration (dark red on the slant) only. Bacteria are strict aerobes.
    A/A; gas Fermentation of all three sugars with CO2 produced. Bacteria are facultative anaerobes.
    K/A; H2S Aerobic respiration (dark red on slant), fermentation of glucose (acid only in butt), anaerobic respiration (black in butt). Bacteria are facultative anaerobes.
    K/A Aerobic respiration (dark red on the slant); fermentation of glucose (acid only in butt). Bacteria are facultative anaerobes.

    How would you interpret the outcome of the TSI slant, the appearance of which is described below?

    Appearance Outcome and Interpretation
    Slant is a dark red color, butt is yellow with noticeable cracking and bubbling.  

    This page titled 4.7: Triple Sugar Iron Medium is shared under a CC BY 4.0 license and was authored, remixed, and/or curated by Jonathon Van Gray via source content that was edited to the style and standards of the LibreTexts platform.