A semi-solid culture medium used to detect: • sulfide formation • indole production • motility  Used for the for

the identification of

Enterobacteriaceae

Procedures:

Stab the Sulfide-Indole-Motility (SIM) medium with the organism once with a straight wire needle to the bottom of the tube Incubate at 37 C for 24 hours. Read the motility before adding Ehlichs / Kovac’s reagent.

Observations:

A. Hydrogen sulfide production

The sulfide reaction is indicated by blackening of the medium along the line of inoculation or in the bottom of the tube  the reaction is derived from organisms bearing sulfur-containing amino acids  Hydrogen sulfide reacts with ferrous sulfate to form ferrous sulfide which is seen as a black colored substance  (+) H S organisms are always motile 2

B. Motility Determination
POSITIVE TEST (MOTILE)  motile organisms migrate from the stab line and diffuse into the medium, causing turbidity  They may exhibit fuzzy streaks of growth at the stab line NEGATIVE TEST (NON-MOTILE)  bacterial growth accentuated along stab line, surrounding mediums remains clear CONTROL MEDIUM (Uninoculated)  no growth, medium remains clear

C. Indole Production  Add about 13drops of Ehrlichs / Kovac’s reagent to the tube POSITIVE TEST – a red or purple ring indicates that indole is produced at the junction of the medium and the reagent   NEGATIVE TEST – a yellow color is observed.

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