Motility Test –

Expected Results
 Positive: Diffuse, hazy growths that spread throughout the
medium rendering it slightly opaque.
 Negative: Growth that is confined to the stab-line, with sharply
defined margins and leaving the surrounding medium clearly
transparent.
Method
1. Touch a straight needle to a colony of a young (18- to 24-hour)
culture growing on agar medium.
2. Stab once to a depth of only 1/3 to ½ inch in the middle of the
tube. Be sure to keep the needle in the same line it entered as it is
removed from the medium.
3. Incubate at 35°-37°C and examine daily for up to 7 days.
4. Observe for a diffuse zone of growth flaring out from the line of
inoculation.

Uses
 It is used for the differentiation of microorganisms on the basis of
motility in a laboratory setting.
 It is performed to assign taxonomic classification to organisms.
 Motility tests are important in characterization of pathogens.
 The tests are often employed in identification protocols in the
family Enterobacteriaceae
 Motility test is also used for the species differentiation of gram
positive cocci, Enterococci.  Enterococcus faecium and E.
faecalis are non-motile, whereas E. gallinarum and E.
casseliflavus/E. flavescens generally are motile.
Motility is the ability of an organism to move by itself by means of propeller-
like flagella unique to bacteria or by special fibrils that produce a gliding form
of motility. Motile  bacteria  move  using   flagella, thread like  locomotor 
appendages  extending  outward  from  the plasma membrane and cell wall
either single flagellum or multiple flagella. Motility has long been recognized
as an important taxonomic tool and biological characteristic of
microorganisms. The presence of flagella occurs primarily in bacilli but there
are a few flagellated cocci, thus motility is a very important means of
identification in the family Enterobacteriaceae. From the early days in the field
of microbiology, the ability of bacteria to move has been used as a means of
differentiation and classification of organisms.

Objective
 To determine the motility of bacterium.
 To differentiate between motile and non motile bacteria.
Principle
Motility by bacterium is mostly demonstrated in a semi solid agar medium. In
semi-solid agar media, motile bacteria ‘swarm’ and give a diffuse spreading
growth that is easily recognized by the naked eye.The medium mainly used for
this purpose is SIM medium (Sulphide Indole Motility medium) which is a
combination differential medium that tests three different parameters, Sulfur
Reduction, Indole Production and Motility. This media has a very soft
consistency that allows motile bacteria to migrate readily through them
causing cloudiness. The inoculum is stabbed into the center of a semisolid
agar deep. Bacterial motility is evident by a diffuse zone of growth extending
out from the line of inoculation. Some organisms grow throughout the entire
medium, whereas others show small areas or nodules that grow out from the
line of inoculation. The non-motile bacteria will only grow in the soft agar
tube and only the area where they are inoculated.