MOTILITY

DEFINITION – Motility is defined as true movement of bacteria or other microorganism due to its organ of
locomotion which is flagella and in some cases (spirochetes) it is the function of the whole body .

METHODS OF DETECTING MOTILITY

A) MICROSCOPIC METHOD B) NON-MICROSCOPIC METHOD

a) Hanging drop method a) Semi – solid agar media
b) Examination of unstained wet mount b) Solid agar media
c) Warm stage method c) Flagellar staining method
d) Phase contrast microscope
e) Dark ground microscope

A) MICROSCOPIC METHOD – Unstained preparation of living organisms are required for detecting
motility of a given specimen of bacterial culture. Important to note during microscopy is defocusing
of condenser to minimize the contrast between organism and its background, though it may cause loss
of some resolution.

a) HANGING DROP METHOD

PROCEDURE :

1. A glass slide is cleaned with cotton and is made grease free by passing it over the flame.
2. A clean disposable coverslip is taken and is made grease free
3. Then small pillars of paraffin or vaseline are constructed on the corners of all along the edges and
borders of coverslip with a small glass rod
4. Now a drop of normal saline is taken on the coverslip and a small colony is taken from the plate by
using a sterile loop and an emulsion is made carefully over the coverslip. It is essential not to spread
the emulsion.
5. Once the suspension is ready the coverslip is quickly turned upside down without allowing the drop
to fall.
6. Now the coverslip is mounted over glass slide, then the arrangement is transferred to microscope.
It is then observed first under low power objective to find out the edge of drop. Then switch over to
high power objective to focus on edge. The condenser is defocused by rotating it downward to reduce
and diffuse illumination. This step of defocusing renders the organism more refractile and visible
with the only demerit of losing some resolution.

Advantage of focusing the edge

 O2 concentration is higher at the edge as compared to centre, so organisms try to move to
periphery.
 In the centre, the movement of organism is vertical and no horizontal movement is observed.
And in the edge, the movement is horizontal.
b) EXAMINATION OF UNSTAINED WET FILM – A 6-24 hour broth culture is made. The
suspension is kept between a coverslip above and a clean dry glass slide below. In this method, no
gap is present between suspension and slide.

c) WARM STAGE ILLUMINATION – Warm stage at 37ºC is placed over the main stage of
microscope. This method is useful for protozoa like Entamoeba histolytica. It also helps to see growth
and lysis of bacteria on blocks of nutrient agar.

d) PHASE CONSTRAST MICROSCOPE – For motile organisms like spirochetes.

e) DARK FIELD (OR DARK GROUND) MICROSCOPE – The reflected light is used instead of
transmitted light in this method. Hence dark ground condenser is used. Used to visualize living,
unstained cells and thin bacteria like spirochetes.

B) NON-MICROSCOPIC METHODS

a) SEMI-SOLID AGAR – In such media, motile bacteria swarms and gives a diffuse spreading growth
that is only recognizable by naked eye.

PROCEDURE :
0.2% agar is dissolved in nutrient broth. 10 ml of this is suspended in tubes and left vertically.
Inoculation is done with a straight wire making a simple stab down the centre of tube reaching about
half the depth of media. It is then inoculated under condition favouring motility at 37ºC and examined
at intervals of 6hors, 1, 2 and 6 days.Freshly prepared medium containing 1% glucose can be used for
motility test of anaerobes.
Incorporation of 0.005% tetrazolium chloride helps to see motility. It is colourless in oxidized form
but turns red on reduced state meaning motility.
Inference – Non motile organisms generally show growth confined to straight wire, having sharply
defined margin and keeping the area transparent.Motile organisms show typical hazy growth reducing
the mediuem opaque.

Craige’s tube method. – It is a subtype of semisolid agar method done for Salmonella. It is a wide
tube containing 0.2% nutrient agar, at the centre of which is embedded a short narrow tube open at
both ends in such a way that one end of it projects over the other. Inoculation is done inside the inner
tube. After incubation subculture is taken from surface of agar outside the centre of tube.

U-tube method – It contains 0.2% agar. Inoculation is done at one end of U shaped tube and then
incubated at 37ºC. Subcultures are taken from other end to check for growth of organisms. The motile
organisms spread to the other end.
b) SOLID AGAR MEDIA – Certain bacteria show swarming growth in solid agar media. Eg. Proteus.
 c) FLAGELLAR STAINING – Done with the help of electron microscopy and special staining
method.

When examining living organisms for property of active locomotion it is essential to distinguish between
true and false motility.

True Motility – Organism will move in different direction and change their position with respect to each
other

False motility can be –

1. Passive drifting : movement of organisms in same direction in a conventional current in the fluid.
2. Brownian movement : Oscillatory movement about nearly a fixed point possessed by small bodies
suspended in fluid. It is due to irregularities in the bombardment with water molecules.

TYPES OF MOTILITY

Vibrio cholera – Shooting star / Darting type due to polar monotrichous flagella
Salmonella – Tumbling motility due to peritrichious flagella
E.coli – Sluggish motility
Treponema pallidium – exhibit rotator or cork-screw movement due to endoflagella
Clostridium – Stately motile except C. perfringes
Borellia vincentis – Coarse lashing motility
Borellia recurrentis – Rotatory/Oscillatory movement
Giardia , Trichomonas - Falling leaf like
E.histolytica – Gliding movement
Bacillus, Proteus, Pseudomonas, Helicobacter, Campylobacter – Actively motile

Non motile organisms – Staphylococcus, Streptococcus, Neisseria, Corynebacterium, Shigella, Klebsiella,

Bacillus anthracis, Clostridium perfringes

FUNCTIONS OF MOTILITY

1. Search of nutrients.
2. Help pathogenic bacteria to penetrate through viscid mucous and epithelial barrier to further spread
infection
Though these theories are there but against this theory, Brucella, despite being non motile is capable of
causing invasive disease.