Diagnostic bacteriology II
Diagnosis and characteristics of Streptococcus spp.
3-2-2025
Learning outcomes
• Define streptococcus.
• Demonstrate knowledge about its types.
• Learn how to diagnose them
• Use different method for their identification.
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General characteristic
• Gram positive.
• Cocci, arranged in chains or pairs.
• Non-motile.
• Non-spore forming.
• Facultative anaerobes.
• Some species can form capsules.
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Classification of streptococci based on many features including:
1. Oxygen requirement (aerobes and obligate anaerobes).
2. Hemolytic properties on blood agar (alpha, beta, and gamma).
3. Antigenic structure (Lancefield classification) for beta hemolysis.
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• Medically important streptococcus are:
1. Streptococcus pneumonia (alpha hemolysis)
2. Viridians streptococci (alpha hemolysis)
3. Streptococcus pyogenes (beta hemolysis + group A Lancefield classification)
4. Streptococcus agalactiae (beta hemolysis + group B Lancefield classification)
5. Enterococcus sp. (no hemolysis- gamma )
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• Streptococcus pneumonia
• Pneumonia (60-70% of bacterial pneumonia), sinusitis, otitis media, septic arthritis
and meningitis.
• The onset of pneumococcal pneumonia is usually sudden with fever, chills & sharp
pleural pain, and rusty sputum.
• Early in the disease bacteremia is present in 10-20% of cases.
• The mortality rate may reach up to 30% depending on age and underlying illness.
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Viridans streptococci
• Including: S. mitis, S. mutans, S. salvaris, S. sanguis and others.
• S. mutans :commensal flora of oral cavity.
• They can cause a Varity of infections such as dental caries and subacute bacterial
endocarditis.
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Streptococcus pyogenes
• Erysipelas, cellulitis, necrotizing fasciitis (flesh-eating), puerperal fever, sepsis,
streptococcal sore throat (s. pyogenes is the most common bacterial cause of sore
throat), streptococcal pyoderma (impetigo).
• Streptococcal toxic shock syndrome: Following minor trauma, characterized by
fever, bacteremia, rapidly progressive shock, and multi-organ failure.( blood culture
are often positive).
• Scarlet fever: it can develop in people who have strep throat. It's characterized by a
bright red rash usually on trunk and extremities, fever and sore throat.
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• Streptococcus agalactiae
• Present as normal vaginal flora in 5-25% of women.
• Causes neonatal sepsis and meningitis.
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Enterococcus sp.
• There are at least 12 species: Enterococcus faecalis is the most common and
causes 85-90% of enterococcal infections.
• It is known to cause nosocomial infections particularly among
immunocompromised patients.
• Enterococcal infections include UTI, wound infections, biliary tract and blood.
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Sample collection
• Urine
• Pus
• CSF
• Biological fluids
• Blood
• Throat swab
• Cervical swab
• Sputum
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Microscopic examination-gram stain
Microscopically the observation will be as follows:
• Gram positive bacteria
• Cocci in shape arranged as chains or diplococci.
• Purple in color
• Capsule observed in some species.
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Culture characteristic
• Alpha hemolysis
Streptococcus pneumonia a small round alpha hemolytic colonies at first dome-
shape and later develop a central plateau with an elevated rim.
Viridans streptococci It produce alpha-hemolysis on blood agar, Convex shaped
colony.
• Beta hemolysis
Streptococcus pyogenes colonies are usually compact, small, and surrounded by a 2-
to 3-mm zone of β-hemolysis.
Streptococcus agalactiae smooth convex colonies.
• Gamma hemolysis
Enterococcus sp. (Enterococcus faecalis) no hemolysis on blood agar, small
(pinpoint colonies), smooth, gray or grayish-white.
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S. pneumonia
S. Pyogenes
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Identification methods
• Catalase test Negative for catalase test.
• Bacitracin sensitivity
To distinguish between S. pyogenes (susceptible to B) & non group A such as S.
agalactiae (Resistant to B)
Bacitracin will inhibit the growth of group A S. pyogenes giving zone of inhibition
around the disk.
• Optochin sensitivity
S. pneumonia is sensitive while viridis streptococcus are resistant.
• Bile solubility test
S. pneumoniae produce a self-lysing enzyme to inhibit the growth.
The presence of bile salt accelerate this process.
Positive test appears as clearing in the presence of bile while negative test appears
as turbid. (S. pneumonia (+ve ) Viridans streptococcus (-ve))
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• Anti- Streptolysin O Test (ASOT)
ASOT this test is serological test used in post streptococcal infection complications
depending on the presence of anti-streptolysin-O antibody in the blood of patient’s
previously infected with S. pyogenes.
normal < 100< positive result.
• Bile-esculin test
Bile-esculin test is based on the ability of certain bacteria, notably the group D
streptococci and Enterococcus species, to hydrolyze esculin in the presence of bile
(4% bile salts or 40% bile).
1. Positive : Enterococcus species (e.g. E. faecalis)
2. Negative : Viridans streptococcus, not group D
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Throat swab for pharyngitis
1. Ask the patient to open their mouth and say “AH”.
2. Gently depress the tongue with a tongue depressor.
3. Guide the swab over the tongue to the posterior pharynx.
4. Gently swab the mucosa behind the uvula and between
the tonsillar pillars back and forth.
5. Remove the swab without touching the tongue, uvula, or lips.
6. The swab is cultured on blood, chocolate and MacConkey agar and incubated for 24H at
37℃
7. Gram stain and observe microscopically for gram positive cocci .
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