Streptococcus spp
Assist. Prof. Rajeshwar Ray
Medical Microbiologist
NHPC Reg. : A-71 Diagnostic Med. Microbiology
Streptococcus spp
History
• Bilroth in 1874: He firstly described that cocci are arranged in chain in
purulent exudates from erysipelas and wound infection and gave the name
Streptococci.
• Rosenbach in 1884 gave the name Streptococcus pyogenes to cocci
arranged in chain which he isolated from human suppurative lesions.
Morphology
• Gram +ve cocci in chain.
• Length of chain depends in culture condition. In liquid medium, chain is
longer in comparison to solid medium.
• Longest chain: Streptococcus salivarius (Non-pathogenic)
• Shortest chain : Diplococci (Streptococcus pneumoniae)
Con…..
• Short chain: Streptococcus pyogen etc
• Non-motile
• Non-sporing
• Non-Acidfast.
• Capsulated:
Group A and C: Possess Hyaluronic acid capsule
Group B and D: Possess polysaccharide capsule.
-Noticeable in young culture i.e. log phage but not in stationary phage.
Streptococcus in chains (Gram stain)
5
Streptococcus pneumoniae (diplococcus). Fluorescent stain
6
Culture properties
• Aerobe & Facultative anaerobe
• Obligate anarobe: Peptostreptococcus spp
• Opt.temp. - 37 oC (22 oC)
• Opt. PH -7.4
• Do not grow on ordinary media
• Growth occurs only in enriched media
• On Serum broth
– Produce granular turbidity with powdered deposit
– No pellicle is formed
Classification on the basis of O2 requirement, Hemolysis and Lancefield type.
Streptococcus spp
Aerobe and Facultative Anaerobe Obligate Anaerobe : Peptostreptococci
Hemolysis
Classification
Streptococci
Alfa hemolytic
Beta hemolytic Gamma hemolytic
20 Lancefield groups
Group A
80 Griffith types
Hemolysis reaction - sheep blood agar
: Brown classification of Streptococci on the basis of Haemolysis in 1919
A. α (alpha): Viridans group and Streptococcus pneumoniae :
- Cause opportunistic infection
- partial hemolysis : Zone size- 1-2 mm , green color
B. β (beta) : Complete clear Zone: Zone size- 2-4 mm
- Pathogenic Organism
C. γ (gamma): Enterococcus spp and Streptococcus faecalis.
- Usually Non-pathogenic, No hemolysis
Note: Viridan and Enterococcus group classified into species by physiological and biochemical properties.
Hemolysis
• Groups A and B : Beta Hemolysis
• Group D : α or γ Hemolysis : S. pneumoniae and viridans
Streptococci
Hemolysis on Blood agar
-hemolysis
-hemolysis
-hemolysis
Serological classification
1. Group Specific polysaccharide antigen of cell wall.
• Based on differences in Group specific polysaccharide antigen of cell wall,
Lancefield in 1933 classified Most strain of beta Hemolytic and some strain of
alpha and gamma Hemolytic streptococci are classified into 20 Lancefield
Groups.
• According Lancefield: Streptococci classified from A through H, K through V.
i.e. ABCDEFGHKLMNOPQRSTUV.
• Majority of streptococci causing human infection are beta-haemolytic and
belong to Group A which are given the species name; Streptococcus
pyogenes.
• Other serious human pathogens fall into Group B,C,D and G.
Con…..
2. Type specific antigen:
Based on type specific antigen; M, T and R, strain of group A (S. pyogenes) is
further subdivided into 80 Griffith serotypes (type 1, 2 …etc).
• M protein is important type specific antigen which exits approx. 80 different
antigenic form, each of which is present in a different serotype of S.
pyogenes (type 1, type 2 , type 3 etc)
• Serotyping is useful in epidemiological study.
Biochemical reaction
• Catalase- -ve
• Oxidase- -ve
• Ferment several sugars producing acid but no gas
- Major fermentative product is lactic acid which accumulates in culture media that rapidly terminates bacterial
growth.
- Sugar serum peptone waters should be employed for fermentation test.
• Donot liquefy gelatin
• Ribose fermentation : -Ve
• PYRase test are useful in distinguising S. pyogenes from non Group A haemolytic
Streptococci.
Diagnosis
• Clinical evidence
• Laboratory evidence
Laboratory Diagnosis
A. Microscopy
B. Culture properties
C. Biochemical properties
D. Serological diagnosis
Sample : Pus, Sputum, Fluid
Direct Gram Stain Inoculate in BA/MA sometime CA (e.g. sputum)
Observe Colony characters
Gram +ve Gram -Ve
Cluster
Chain Diplococci
Catalase +ve
Streptococcus spp
Staphylococcus spp
Coagulase test
Coagulase test: +ve Coagulase test: -Ve
S. aureus, S. Intermedius, S. hyicus S. epidermidis, S. saphrophyticus
Hemolysis
Manitol +ve : S. aureus
Hemolysis –ve : S. hyicus Hemolysis + ve
S. aureus, S. intermedius
Manitol –Ve : S. intermedius
Streptococcus spp
Hemolytic on BA
No Hemolytic (-Ve) : Various streptococci
Non-pathogenic Hemolytic (+Ve), S. equi, S. agalactiae, S. pneumoniae
CAMP test
- ve : S. equi, S. pneumoniae
+ve , S. agalactiae,
Optochin : Sensitive Optochin: Resistance
Streptococcus pneumoniae Streptococcus equi
Optochin : R
Bacitracin: R
Viridans group
Optochin : S
Bacitracin: R
Streptococcus pneumoniae
Optochin: R
Bacitracin: S
Streptococcus pyogene
