Microbiology

Bpt 3rd semester

 Pneumococci
• Pneumococci are normal commensals of the upper
respiratory tract.
• They are important pathogens of pneumonia and
otitis media (infection in middle ear) in children.
• They are reclassified as Streptococcus pneumoniae
because of its genetic relatedness to streptococcus.
• They differ from streptococci in their morphology
( diplococci),bile solubility, optochin sensitivity
and by a specific polysaccharide capsule.
• A. Morphology
• Pneumococci are Gram positive, small (1 μm diameter),
slightly elongated cocci arranged in pairs (diplococci) with
the broad ends in apposition.
• Each coccus has one end broad or rounded and other
pointed (flame shaped or lanceolate appearance)
• They are capsulated and the capsule encloses each pair. The
capsule may be demonstrated as a clear halo in India ink
preparation
In old cultures, the capsule is usually lost.
• They are non-motile and non-sporing.
• Pneumococci have complex nutritional requirement and
therefore grow only in enriched media especially
supplemented with blood.
• They are aerobes and facultative anaerobes and their
growth is improved by 5- 10 per cent CO2.
• The optimum temperature for growth is 37°C (range 25°C
to 42°C) and pH 7.8 (range 6.5 - 8.3).
• On blood agar, after incubation for 18 hours, the colonies
are usually small (0.5- 1 mm) , dome shaped, with an area
of greenish discolouration (alpha haemolysis) around
them.
• Pneumococci are typically alpha haemolytic but under
anaerobic conditions colonies show beta haemolysis due
to liberation of oxygen labile pneumolysin O by these
bacteria
• In liquid medium such as glucose broth, pneumococci
produce uniform turbidity.
• These cocci readily undergo autolysis in cultures due to
the action of intracellular enzymes. Autolysis is enhanced
by bile salts and other surface active agents. This
property is helpful to differentiate it from other
streptococci.
• Pathogenesis
• Str. pneumoniae is one of the most common bacteria
causing pneumonia, both lobar and bronchopneumonia.
• It is also responsible for acute tracheobronchitis and
empyema.
• 1. Lobar Pneumonia
• Pneumonia results only when the general resistance is
lowered. Common infective types of Str. pneumoniae
include types 1- 12 in adults and types 6, 14, 19 and 23
are responsible in children.
• Bile solubility test is an important diagnostic test to differentiate
pneumococcus from other streptococci. S.pneumoniae are bile esculin positive.
• This test is based on the presence of an autolytic enzyme amidase in the
pneumococci .
• This enzyme cleaves the bond between alanine and muramic acid in the
peptidoglycan.
• The amidase is activated by bile salts, resulting in lysis of the bacteria.
• Pneumococci are catalase and oxidase negative.
• Pneumococci are sensitive to optochin (ethy lhydrocuprein hydrochloride) in a
concentration of 1/500,000. When a 6 mm optochin disc (5 μg) is applied on a
blood agar plate inoculated with pneumococci, a wide zone (14 mm or more)
of inhibition occurs on incubation.
• This is very useful test to differentiate pneumococci from other streptococci
which do not show zone of inhibition by optochin disc.
• 3. Meningitis
• It is the most serious of pneumococcal infections. Str.pneumoniae is
the second most important cause of pyogenic meningitis after
N.meningitidis. This disease is commoner in children.
• Pneumococcus spreads from the pharynx to the meninges via blood
stream.
• Other bacterialagents of pyogenic meningitis include N meningitidis,
H. influen zae, Str. agalactiae (group-B ) and Listeria
monocytogenes.
• 4. Other Infections
• Pneumococcus may also produce empyema, pericarditis,otitis media,
sinusitis, conjunctivitis, peritonitis and suppurative arthritis, usually
as complications of pneumonia.
• Transmission:Pneumococci are transmitted by inhalation
of contaminated dust, droplets or droplet nuclei.
• Infection usually leads to pharyngeal carriage. Disease
results only when the host resistance is lowered by
factors such as respiratory viral infections, malnutrition
etc.
• LobLobar pneumonia is usually a sporadic disease but
epidemics may occur. The incidence of
bronchopneumonia is more when an epidemic of
influenza or other viral infections of the respiratory tract
occurs.
 1. Specimens
 Clinical samples, such as sputum, cerebrospinal fluid (CSF),
pleural exudate or blood are collected according to the site of
lesion.
 Blood culture is useful in pneumococcal septicaemia.
 2. Collection and Transport
 All the specimens should be collected in sterile containers
under all aseptic conditions.
 They should be processed immediately.
 In case of delay, CSF specimen should never be refrigerated but
kept at 37°C (H. influen zae, another causative agent of pyogenic
meningitis may die at cold temperature).
• 3. Direct Microscopy and Antigen Detection
• Gram staining of smear reveals a large number of
• polymorphs and typical organism. In case of
meningitis, presumptive diagnosis may be made
by finding Gram positive diplococci which may be
intracellular as well as extracellular in CSP smear.
• Capsule swelling may be observed under
microscope, when pneumococci are mixed with
type specific antisera.
• 4. Culture
• Specimen is inoculated on blood agar and incubated at 37°C for 24 hours
in the presence of 5- 10% CO2.
• Typical colonies develop with a - haemolysis. Organisms from the
isolated colony are identified by Gram staining and biochemical
reactions.
• 5. Colony Morphology and Staining
• Colonies are usually small (0.5 - 1 mm), with alpha haemolysis around
them.
• On Gram staining pneumococci are Gram positive, small (1 μm
diameter), diplococci, They are flame shaped or lanceolate in appearance.
• The capsule may be demonstrated as a clear halo in India ink preparation.
 Treatment

• Treatment
• The antibiotic of choice is parenteral penicillin.
• Cephalosporin is indicated in case of penicillin resistant strains.
• A polysaccharide vaccine containing prevalent serotypes (23
serotypes) is administered by a single dose injection .
• Such vaccines are used only in those persons who are at enhanced
risk of pneumococcal infection.
• These include persons with absent or dysfunctional spleen,
nephrotic syndrome, sickle cell anaemia, multiple myeloma ,
hepatic cirrhosis, diabetes mellitus and immunodeficiencies
including HIV infection. Vaccination is contraindicated in
lymphomas.