CAPSULE OF THE

BACTERIAL CELL

D-r Rozalina Yordanova

Department of Microbiology
 CAPSULES AND SLIME LAYERS

• Structures surrounding the outside of the cell

envelope
• If it is too thin, organized and is firmly attached to
the cell wall, it is called microcapsule.
• If it is loose and many cells are embedded in a matrix
it is known as slime.
• Not essential to cell viability
• Often lost during in vitro culture
• Usually consist of polysaccharide; however in
certain Bacillus anthracis they are composed
of a polypeptide (polyglutamic acid)

• The bacteria of a single species can be

classified in different capsular serovars (or
serotypes) based on the fine chemical
structure of this polysaccharide
 FUNCTIONS
• Attachment
• Protection from phagocytosis
• Resistance to drying
• Depot for waste products
• Reservoir for certain nutrients
• Protection
• Resistant to washing off
• Usually weakly antigenic
• Endows virulence
THE CAPSULE OR SLIME LAYER
• mediates adherence of
bacteria to human tissue
• determines virulence of
many bacteria protecting
bacterial cells from
engulfment by phagocytes
• protects cells from
perennial effects of
desiccation
• determines antigen
properties of bacterium
and can be used for
serologic identification
 Capsules, Slime Layers, and S-Layers

• Some procaryotes have a layer of material lying outside the cell wall. This
layer has different names depending on its characteristics. When the layer
is well organized and not easily washed off, it is called a capsule. It is
called a slime layer when it is a zone of diffuse, unorganized material that
is removed easily. When the layer consists of a network of
polysaccharides extending from the surface of the cell, it is referred to as
the glycocalyx, a term that can encompass both capsules and slime layers
because they usually are composed of polysaccharides.
• However, some slime layers and capsules are constructed of other
materials. For example, Bacillus anthracis has a proteinaceous capsule
composed of poly-D-glutamic acid.
• Capsules are clearly visible in the light microscope when negative stains or
special capsule stains are employed; they also can be studied with the
electron microscope
 BIOFILM

• A bacterial biofilm is a structured community of

bacterial cells embedded in a self-produced polymer
matrix and attached to either an inert surface or
living tissue
• Such films can develop considerable thickness (mm)
• The bacteria located deep within such a biofilm
structure are effectively isolated from immune
system cells, antibodies, and antibiotics. The
polymers they secrete are frequently glycosides,
from which the term glycocalyx (glycoside cup) for
the matrix is derived.
 EXAMPLES OF MEDICALLY IMPORTANT BIOFILMS

1. Streptococcus mutans, which is a bacterium that

colonizes teeth and excretes acid that causes tooth
decay. Normally the body’s immune system would
surround the bacterium and eventually kill it, but that
doesn’t happen with Streptococcus mutans. It has a
capsule, which prevents the Streptococcus mutans
from being recognized as a foreign microorganism by
the body’s immune system. This results in cavities

2. Oral streptococci and other bacteria attach to the

surface of the cardiac valves to form a biofilm.
Professional phagocytes are attracted to the site and
attempt, unsuccessfully, to phagocytize the bacteria.
The frustrated phagocytes then release the tissue-
damaging content of their lysosomes resulting in an
inflammatory reaction and the clinical picture of
endocarditis
 CAPSULE STAINING
METHODS:

• For observing of capsule special staining may be used

1) Negative staining

2) Positive staining

3) Mc Fdyean reaction: which uses the Loeffler

polychrome methylene blue to demonstrate the
capsule of the Bacillus anthracis
For positive staining of smears:
• Make a smear from colony of S. pneumoniae on a clean grease free glass slide,
and allow it to air dry. Note: The smear should not be heat fixed.
• Put the smear on a slide rack and flood smear with crystal violet. Allow it to stain
for 5-7 minutes.
• Wash the smear with 20% copper sulphate solution and blot it dry.

• Observe the smear first under low power (10x) objective, and then under oil
immersion(100x) objective.
• In the culture smear, the capsule is seen as a light blue in contrast to the deep
purple colour of the cell.
For negative staining of smears:
• Take a clean grease free glass slide.
• Put a large loop full of undiluted India ink on the slide.
• Then add a small loop full of liquid bacterial culture to the India ink and
emulsify.
• Take a clean, grease free cover slip and place on the ink drop and press
it down, so that the film becomes very thin and thus pale in colour.
• Observe the wet film under high power (40x) objective.
• The capsule in negative staining method is seen as clear refractile, halo
around the organism against a black background.
• Capsules are most easily seen by negative staining,
outlined as a light area against a darkened background

Negative staining technique. The capsules appear as a clear halos zone between the
colored background and the stained cells.
 CAPSULE STAINING: Microscopy

Capsulated Bacteria In India ink Cryptococcus neoformans in India ink