BACTERIA

Bacteria can be classified according to their

shape into:

1 – Spherical (cocci)
2 – Rods (bacilli)
3 – Comma shaped (vibrios)
4 – Spirals (spirochetes)
 COCCI
G+ve G-ve

Groups: Staphylo-cocci Neisseria

Chains: Strepto-cocci
Diplo : Strept. pneumoniae
COCCI

Streptococci
Gram – positive spherical or ovoid cells,
Arranged in chains or pairs (due to division
in single plane).
All species are catalase negative.
Their growth requires enriched medium
containing blood or serum.
STREPTOCOCCUS
 STREPTOCOCCUS
• Streptococci are gram-positive spherical or ovoid cells,
arranged in chains or pairs due to division in single
plane.

• All species are catalase negative.

• Their growth requires enriched medium containing

blood or serum e.g., sheep blood. Streptococcus
pyogenes, and S. pneumoniae are the most important
species.
Streptococci

Group carbohydrate antigens

The streptococcal cell wall contains
several forms of polysaccharides, one of
which constitutes the Lancefield group
antigen.
It is detected serologically using specific
antibodies.
Streptococci: CLASSIFICATION

1 – Haemolytic reactions on sheep blood agar

2 – Colony size
3 – Lancefield serological groups
4 – Biochemical reactions (carbohydrate
fermentation pattern)
5 – DNA-DNA hybridization
6 – 16S r RNA (a current classification of the
genus Streptococcus in 6 groups).
 Streptococci: CLASSIFICATION

On basis of the haemolytic action

on blood agar, Streptococci are
divided into 3 main types:
I – Beta haemolytics (colonies are
surrounded by complete
haemolysis of RBCs)
II – Alpha haemolytics (colonies
are surrounded by greenish
discoloration of the media =
partial or incomplete
haemolysis of RBCs)
III – Gamma haemolytics (no
change of the media = no
haemolysis of RBCs)
STREPTOCOCCUS PYOGENES
Group A beta-hemolytic
streptococcus
 Pathogenesis and virulence factors
Streptococcus pyogenes owes its major success as a
pathogen to its ability to colonize and rapidly multiply
and spread in its host
1. M protein: It is an anti-phagocytic cell surface proteins ,
It is immunogenic.
2. Fibronectin-binding protein (Protein F) and lipoteichoic
acid (LTA) for adherence.
3. C5a peptidase: A protease that cleaves C5a
4. Hyaluronic acid capsule acts as an immunological mask
to avoid phagocytosis. Hyaluronic acid is chemically
similar to that of host connective tissue. Therefore, it is
not immunogenic.
5. Invasins:
a- Streptokinase: It activates plasminogen of human plasma into
plasmin that digests fibrin and fibrinogen. Streptokinase (SK) has been
referred to as fibrinolysin or streptococcal spreading factor.
b- Hyaluronidase: It can digest host hyaluronic acid. Thus helping
spread of infection in tissues.
c- Nucleases: There are DNases (Streptodornases).
d- Streptolysins: streptolysin O (oxygen labile) and streptolysin S
(oxygen stable). Streptolysin O (SLO) is a highly immunogenic protein.
Streptolysin S (SLS) is responsible for the β-haemolysis around the
colonies of the organism. It is non-immunogenic.

6. Pyrogenic exotoxins: These toxins act as superantigens Three

different streptococcal pyrogenic exotoxins (SPE A, B and C) have been
identified. SPE A is also referred to as erythrogenic toxin A or scarlet fever
toxin because it is responsible for the erythematous rash characteristic of
this disease. SPE B acts as a protease. These toxins cause toxic shock
syndrome, and necrotizing fasciitis.
 DISEASES CAUSED BY S. PYOGENES
1. Pharyngitis (sore throat, tonsillitis)
2. Scarlet fever: caused by an erythrogenic toxin-producing S. pyogenes.
3. Skin and soft tissue infections
•Impetigo (pyoderma): It is a local infection of the superficial layers of
the skin with blisters and denuded surface covered with crusts.
•Cellulitis and erysipelas: Cellulitis is infection of the deep layers of the
skin Erysipelas is a form of cellulitis accompanied by fever and systemic
toxicity.
4. Invasive streptococcal infections
•Puerperal fever.
•Acute endocarditis: The condition can occur in individuals with normal
or damaged heart valves.
•Necrotizing fasciitis is an invasive disease associated with severe
tissue destruction particularly associated with SPE B
•Toxic shock syndrome: This severe form of invasive infection often
begins with skin wounds or minor traumas and rapidly deteriorates
leading to necrotizing fasciitis or myositis. Shock, renal failure and
acute respiratory distress syndrome are complications of the condition.
5. Post-streptococcal sequelae:
1. Acute rheumatic fever (ARF)
2. Acute glomerulonephritis (AGN)

These conditions are non-suppurative

and are based on immune-mediated
mechanisms.
 Tonsillitis
Necrotizing Fasciitis

Impetigo
 Laboratory diagnosis
A-Samples: A throat swab, pus, high vaginal swab. Blood for Blood
culture in case of bacteraemia.
B-Direct detection
•Gram-stained smear: Gram-positive cocci in pairs or chains.
•Streptococcal group A antigen in throat specimens.
C-Cultivation
Specimens are plated directly onto sheep blood agar and incubated at
37oC with 5% CO2.
Blood culture should be done in cases of bacteraemia accompanying
infections like puerperal sepsis and endocarditis.
D-Identification
- Colonies showing catalase negative Gram-positive cocci in pairs or
chains can be considered Streptococcus species for further identification.
- Colonies surrounded with wide zone of beta-haemolysis should be
tested for serogroups and bacitracin susceptibility. S. pyogenes belongs
to group A and is susceptible to bacitracin.
Gram stained smear of Streptococcus infection
Streptococcal Culture on Blood Agar

Beta hemolysis on blood agar

 Identification of streptococii

Gram stained smear of Streptococcal culture

Identification of streptococii
 Serological tests
Anti-streptolysin O (ASO) is the antibody response most often
examined to confirm previous S. pyogenes infection in a suspected
case of ARF, when S. pyogenes could not be isolated from the site of
infection.
1. An elevated ASO titre above 200 units.

2. anti-DNase B or anti-streptokinase A value above 80 units is of

significance for either of these 2 tests.

3. high levels of C-reactive protein (CRP) and Erythrocyte sedimentation

rate (ESR) help in diagnosis and follow up of ARF cases.
 Treatment and prevention

Penicillin is still uniformly effective in

treatment of S. pyogenes disease. Long
acting penicillin is used also as a
chemoprophylactic agent against
recurrent S. pyogenes infection to
prevent repeated rheumatic attacks
 Alpha-haemolytic streptococci
VIRIDANS STREPTOCOCCI
They are normal inhabitants of the oral cavity, gastrointestinal tract, and
female genital tract.

•The most serious infection caused by them is subacute bacterial

endocarditis (SBE). It may occur when dental manipulations or trauma
to mucosa of upper respiratory tract, e.g. tonsillectomy, lead to
bacteraemia. The organism settles on the prosthetic valve or the
deformed heart valves e.g. rheumatic or congenital heart. It have also a
significant role in dental plaque formation and dental caries.

•Laboratory diagnosis of SBE: Blood culture and identification of the

isolate from the subculture on sheep blood agar.

•Treatment and prevention

The use of the synergistic combination of penicillin and gentamicin in
life-threatening infections, such as endocarditis, is essential. A single
large dose of ampicillin or amoxicillin should be given to patients with
congenital or rheumatic heart prior to dental procedures to prevent
endocarditis.
STREPTOCOCCUS PNEUMONIAE
PNEUMOCOCCUS.
 STREPTOCOCCUS PNEUMONIAE
PNEUMOCOCCUS

Morphology
Gram-positive, lancet-shaped diplococci, capsulated and
may form short chains. The capsule appears as an
unstained zone around the organism

Pneumococcal capsule: It is polysaccharide, haptinic and

constitutes the major virulence factor of S. pneumoniae; it
is antiphagocytic. There are 90 serotypes, and antibodies
to the capsule confer type-specific protection.
S. Pneumonae Fluorescent antibody (FA) stain
 Pneumococcal diseases

Three major diseases; namely pneumonia, meningitis and

otitis media are caused by S. pneumoniae. Other infections
include sinusitis, conjunctivitis, endocarditis and septic
pericarditis. Individuals at risk of pneumococcal infections
are the alcoholics, post-splenectomy, immunosuppressed,
infants and the elderly.
 Laboratory diagnosis
A)Spcimes: Sputum, CSF, Eye discharge
B) Direct detection
• Gram-stained smear: Gram-positive, lancet-shaped diplococci,
capsulated and may form short chains. The capsule appears as an
unstained zone around the organism
• Quellung test: capsule reacts with the corresponding antiserum and
can be seen under the microscope to swell.
• Detection of capsular polysaccharide antigen in CSF by means of slide
latex agglutination test or ELISA.
C) Culture: Specimens should be plated directly onto sheep blood agar
and incubated at 37oC in 5% CO2. Blood culture should be done in
cases of bacteraemia and endocarditis.
D) Identification: Alpha-haemolytic colonies with depressions in their
centres are characteristic of pneumococci. It is essential to differentiate
between S. pneumoniae and viridans streptococci during sputum
examination.
Gram-stained smear of S.pneumoniae infection
S. Peumoniae culture and identification
S. Peumoniae identification

Quellung test
Differences between S. pneumoniae and viridans streptococci

Test S. Pneumoniae Viridans

streptococci
Optochin susceptibility + -
Bile solubility + -
Mouse virulence + -
Capsular-
polysaccharide-antigen + -

Quellung test + -
DNA –probe-Specific-
to-S.peumoniea + -
Optchin sensitivity test
 ENTEROCOCCUS
•They are Gram-positive cocci that occur singly, in pairs, and in short
chains.
• They grow in broth containing 6.5% NaCl and are able to grow at 45 0C.
•Catalase negative and hydrolyze esculin in the presence of bile salts.
•React with the streptococcal Lancefield group D antiserum.
•Enterococci are frequently resistant to antibiotics. They are absolutely
(intrinsically) resistant to cephalosporins and clindamycin.
•The common 2 species are Enterococcus faecalis and Enterococcus
faecium.

Infections caused by enterococci

-Urinary tract infections are the most common, Intra-abdominal or pelvic
wound infections.
- Bacteraemia., Endocarditis., Abscesses, meningitis, peritonitis,
osteomyelitis, and wound infection.
- Enterococcus faecalis is the species most commonly isolated from
nosocomial rather than community-acquired.