 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.