•
•Streptococci
–Gram positive cocci
–chains or pairs
–catalase negative
–facultative anaerobe
(staphylococci are catalase positive)
Streptococcus in chains
groupablestreptococci
A, B and D
–most important
C, G, F
–rare
hemolysis reaction -sheep blood agar
(alpha)
–partial hemolysis
green color
(beta)
–complete clearing
–A and B
(gamma)
–no lysis
Classification according to hemolysis
Groups A an B
–
Group D
–or
S. pneumoniaeand viridans
–
PNEUMOCOCCUS:
Streptococcus pneumoniae
Introduction
Pneumococci are normal commensals of the
upper respiratory tract
Important pathogen of pneumonia & otitis media
Reclassified as Streptococcus pneumoniae
Differ from Streptococci in morphology, bile
solubility, optochin sensitivity & capsule
MORPHOLOGY
Gram positive cocci
Flame shaped/lanceolate apperance with
one end broad and the other pointed.
They are arranged in pairs (diplococci)
They are capsulated, nonsporing and
nonmotile.
The capsule is visualised with Indian ink
negative staining.
CULTURAL CHARECTERSTICS
Pneumococci are fastidious.
They grows only on enriched media.
They are aerobic and facultative anaerobic.
Grows at optimum temperature of 37º C.
The growth is enhanced by the presence of
5-10% Co2.
Growth on blood agar:
–Colonies are small, dome shaped.
–With area of green discolouration (alpha
hemolysis) around them.
–On further incubation,
Concentric rings are seen on the surface when viewed
from above (draughtsman and carrom coin appearance)
Beta hemolytic Alpha haemolytic streptococci
Colonies on blood agar after 18 hrs incubation
Blood agar plate showing draughtsman colonies
of pneumococcus on prolonged incubation
BIOCHEMICAL REACTION
Catalase negative.
Fermentation of inulinby Pneumococci is
used for differentiating them from other
streptococci as the later does not ferment it.
They are bile soluble.
Optochin sensitive
21
Notoptochin sensitive
optochin sensitive
•Most rapid & most useful: identifies & specifies
type of pneumococci in sputum, spinal fluid,
exudates or culture
•Pneumococcal specimen mixed with (polyvalent)
antipneumococcla serum & methylene blue:
•Positive result: Refractile & swollen capsule under oil
immersion
Quellung Reaction
ANTIGENIC STRUCTURE
The most important antigen of the
Pneumococci is:
–Capsular polysaccharide
–Somatic M protein
–Cell wall cabohydrates(C-substance)
CAPSULAR POLYSACCHRIDES
This is type specific.
Pneumococci are classified into types based on the
nature of capsular polysaccharide.
More than 90 serotypes are recognized.
The virulence of Pneumococci is dependent upon its
capsule which prevents or inhibits phagocytosis.
The antibody of capsular polysaccharide protects
against infection.
M Protein
M Protein is characteristics for each type of
Pneumococci.
It is not associated with virulence and
antibody to M Protein is non protective.
C-Substance
Pneumococci contain a species
carbohydrate antigen which is named as C-
Substance.
It is also detected in blood of patients with
some illness , this is known as C-Reactive
protein (CRP).
TOXINS AND OTHER VIRULENCE
FACTORS.
Pneumococci produce an:
–Hemolysin.
–Leucocidin
-Pneumolysin, is a toxin produced by
Pneumococci.
–It has cytolytic and complement activating
properties.
–It is immunogenic.
PATHOGENESIS
Pneumococcus is one of the most common
bacteria causing pneumonia.
–Lobar pneumonia
–bronchopneumonia
Meningitis
It is most serious of pneumococcal infection.
It is the second most important cause of
pyogenic meningitis afterN.meningitidis.
Common in children.
Spreads from the pharynx to the meninges
via blood stream.
Other infections
Pneumococci may also produce:
–Empyema
–Pericarditis
–Otitis media
–Peritonitis
–suppurative arthritis.
Laboratory Diagnosis
Samples :according to site
Collect and transport
Direct microscopy
Culture and identification
Antibiotic sensitivity
Inhibition by optochin
Growth of colonies of Streptococcus
pneumoniae is inhibited by optochin contained in
the disk applied to the blood agar plate
Lysis by bile salts:
Bile salt such as sodium deoxycholate, dissolves
Streptococcus pneumoniae and clear the
turbidity of the heavy inoculum of organisms
Quellungreaction
Capsule of Streptococcus pneumoniae
swells in the presence of specific
pneumococcal antiserum
Morphology
They are gram positive cocciarranged in pairs.
Lanconateshaped
Capsulated organism
35
GRAM POSITIVE COCCI
S. aureus
Beta hemolytic
mannitol
yellow
+ -
Staphylococcus(Clusters) Streptococcus(pairs & chains)
Catalase
BETA: Bacitracin S.pyogenes(group A)
CAMP/Hippurate S. agalactiae(group B)
Coagulase
S. epidermidis
Non-hemolytic
mannitol
white
ALPHA: Optochin/Bile Solubility S.pneumoniae
GAMMA OR ALPHA: Bile Esculin 6.5% NaCl Group D
Enterococcus
Bile Esculin 6.5% NaCl Group D
Non-Enterococcus
Note: S. viridansis
ALPHA hemolytic and
negative for all the tests
below
+
+
+
++
+
+
-
-
Summary Figure (Identification Scheme)
Hemolysis/Test