Species Lancefie Common
ld Terms
Streptococcus A Group A strep
pyogenes
Streptococci
Streptococcus B Group B strep
agalactiae
General Characteristics:
Streptococcus C Group C strep
▪ Gram positive cocci dysgalactiae
▪ Spherical/round Streptococcus equi
▪ in chains or pairs
Streptococcus bovis D Group D Non
▪ Aerotolerant anaerobe - can
enterococcus
survive even in the presence or in
the absence of oxygen. Streptococcus faecalis D Group D
▪ Catalase negative Streptococcus faecium Enterococcus
▪ Capnophiles / Capnophilic Streptococcus None Pneumococcus
o requires additional 5 - 10% pneumonia
of CO2
o E.x: Strep. Pneumoniae Streptococcus A, C, F, Viridans
▪ Non motile anginosus G, N streptococci
Streptococcus mutans
Cell wall classification: Streptococcus mitis
▪ Based on C polysaccharide (C carbohydrate)
▪ Lancefield grouping
o based in the C polysaccharide substance content of streptococci.
▪ Lancefield grouping can be identified by serological typing with the use of
corresponding antisera.
Identification of lancefield grouping:
▪ Extraction of C substance by addition of antisera.
Brown Classification:
▪ According to hemolytic reaction in blood agar SBA
1. Beta hemolytic
o capable of complete zone of hemolysis
o clear zone around colony
2. Alpha hemolytic
o partial hemolysis
o greenish discoloration
3. Gamma hemolytic
o No hemolysis
o Non hemolytic
4. Alpha Prime Hemolytic or Wide Zone Hemolytic
o no hemolytic reaction around the colony
o intact RBC around colony
o surrounded by wide zone of B hemolysis Viridans
➢ Beta hemolytic:
▪ Streptococcus pyogenes
▪ Streptococcus agalactiae
▪ Streptococcus dysgalactiae
▪ Streptococcus equi
➢ Alpha hemolytic and Gamma hemolytic:
▪ Streptococcus bovis
▪ Streptococcus pneumoniae
➢ Alpha hemolytic, beta hemolytic, and gamma hemolytic:
▪ Streptococcus faecalis
▪ Streptococcus faecium
▪ Streptococcus anginosus
▪ Streptococcus mutans
▪ Streptococcus mitis
Classification of significant species:
I. Streptococcus pyogenes
➢ Virulence factors
1. M protein –
o block Fc portion of IgB
o inhibit phagocytosis
o is equivalent to protein A of Staph. aureus
2. Protein F (Fibronectin binding protein)
o for adherence to host
o epithelial tissue to cause tissue damage
3. Lipoteichoic acid
o tries to mediate attachment to mucosal surface to cause infection
4. Hyaluronic acid capsule
o try to coat bacteria to prevent phagocytosis therefore prevent to be
eliminated or to be killed by phagocytes.
5. Streptodornase ( DNAse)
o equivalent to staphylodornase
o DNAse enzyme that is found in strep pyogenes
o tries to hydrolyze DNA and RNA
6. Streptolysin O
o cause lysis of WBC, RBC, platelets
o oxygen labile
o subsurface, anaerobic
o Immunogenic - increased Ab production
o ASO (anti streptolysin O )
7. Streptolysin S –
o Oxygen stable
o surface, aerobic
o not immunogenic
o responsible of lysing blood cells when incubated
o doesn’t induced Ab production
8. Streptokinase ( Fibrinolysin )
o hydrolyze fibrin clot
9. Hyaluronidase –
o hydrolyze hyaluronic acid
o spreading factor
10. Streptococcal pyrogenic toxin
o Etherogenic
o Exotoxin A – release by bacteria
i. causes scarlet fever of streptococcal Toxic shock syndrome
Different infection produced by Streptococcus pyogenes:
1. Bacterial pharyngitis and tonsillitis
▪ Strep throat / sore throat
▪ red and inflamed tonsils
▪ swollen cervical lymph nodes
2. Pyodermal infection
▪ produce skin infection which are fast producing (may nana / pus)
a. Impetigo
o localized skin infection
o small vesicle, progress to weeping lesion ( may nana / pus)
b. Erysipelas
o common among adult
o acute spreading lesion with intensely erythematous with plainly
demarcated with irregular edge lesion
c. Cellulitic / Cellulitis
o deeper tissue invasion
o peripheral vasculitis or diabetes
o progress to gangrene
d. Scarlet fever
o diffused red rush of chest ( 5-7 days mawawala )
o spread to trunk + extremities
3. Necrotizing fasciitis
o invasive infection with tissue necrosis
o suppurative fasciitis hospital gangrene
o invasive infection w/ inflammation + necrosis of tissue, fascia ,
o flesh eating
4. Streptococcal toxic shock syndrome
o multi organ shutdown, leading to death.
5. Post streptococcal sequelae
a. Rheumatic Heart Fever - cross reactive Ab against Streptococcal Ag in
human heart
(kasi malapit ung throat sa heart kaya nag cacause ng infection sa heart)
b. AGN ( Acute Glomerulonephritis ) - due to the deposition of Ag - Ab
complex in glomerulus (kidney disease)
o Strep O - ASO ( damage due to complement activation )
II. Streptococcus agalactiae
➢ Virulence factor
1. Capsule –
o antiphagocytic activity
2. Diseases
▪ Transmission during delivery
▪ Neonatal sepsis - (infection in blood)
▪ Adult:
▪ Endometritis (Endometrium - sexually damage)
▪ wound infection
III. Group C & G
1. Streptococcus dysgalactiae, S. equisimilis
a. Colony: Resembles S. pyogenes,
▪ larger colony forming b-hemolytic isolates
b. Infection: Pyogenic
2. Streptococcus anginosus (G)
a. Colony: Resembles S. pyogenes
▪ , smaller colony forming b-hemolytic isolates
b. Infection: Pyogenic
IV. Streptococcus pneumoniae
➢ Virulence factor: Capsular Polysaccharide
▪ Antigenic polysaccharide resulting to 80 serogroup
▪ Antiphagocytic
➢ Diseases:
1. Pneumonia (Lobar or Community acquired)
➢ 2. Sinusitis, otitis media, bacteremia, meningitis
Morphology: S. pneumoniae
▪ Gram positive diplococci (Lancet or bullet shape)
▪ Capnophilic - additional 5-10% CO2
➢ Neufeld Reaction (Quellung, capsular swelling)
▪ Procedure: Body fluids + capsular antisera
▪ (+): Capsule swell - under the microscope
V. Viridans streptococci ( A, C, F, G, N )
➢ Virulence factor :-
o Polysaccharide capsule
o Deptrum
o Adhesion
➢ Diseases:
o Subacute Bacterial Endocarditis in damage heart valve
o Bacteremia, sepsis
VI. Enterococcus ( Group D)
➢ S. faecalis, S. Faecium
➢ Virulence:
o Extracellular Surface Protein
o Serine protease
o Gelatinase (For attachment)
➢ Disease:
1. Nosocomial Infection
2. UTI, Bacteremia, Endocarditis
SPECIES Disease Association
S. pyogenes [Link] pharyngitis and tonsillitis
2. Pyodermal Infection
o Impetigo
o Erysipelas
o Cellulitic/Cellulitis
o Scarlet Fever
[Link] fasciitis
4. Streptococcal toxic shock
syndrome
[Link] streptococcal sequelae
A. Rheumatic Heart Fever
B. AGN (Acute
Glumerulonephritis)
S. agalactiae 1. Neonatal Sepsis
[Link]
S. dysgalactiae , S. equisimilis 1. Pyogenic Infection
[Link]
[Link]
S. bovis group 1. UTI
[Link]
[Link] Infections
E. faecalis, E. faecium 1. Nosocomial Infection
2. UTI, Bacteremia, Endocarditis
S. pneumoniae 1. Pneumonia (Lobar or Community
acquired)
2. Sinusitis, otitis media, bacteremia,
meningitis
Anginosus, mutans, mitis [Link] infections,
2. endocarditis,
3. dental caries
6th video
LABORATORY DIAGNOSIS
1. Hemolytic pattern: Brown’s classification
2. Colony characteristics
❖ Group A
➢ Gray white
➢ Pinpoint colony (small)
➢ Large zone of beta hemolysis
❖ Group B
➢ Larger colony
➢ Narrow zone of beta hemolysis
❖ Strep Viridans
➢ Gray
➢ Minute to small colony
➢ Alpha, Beta, Gamma hemolytic reaction
❖ Group D
➢ Cream white
➢ Small colony
➢ Alpha, Beta, Gamma hemolytic reaction
❖ S. pneumoniae
➢ Small, round, glistening, dome shaped umbilicate (old)
➢ Mucoid colony (capsule)
➢ Alpha hemolytic reaction
3. Physiologic Characteristics
➢ Pyogenic
o Pus producing
o Mostly Beta hemolytic
➢ Lactococci
o Group N
o Non hemolytic
o Found in dairy products
➢ Enterococci
o Normal biota of human intestine
➢ Strep Viridans
o Normal Biota in upper respiratory tract
➢ Culture Media
1. Blood Agar – Enriched
2. Blood Agar + Trimethoprim and Sulfamethoxazole – for Beta hemolytic Strep
Biochemical Identification
1. Bacitracin (Taxo A)
▪ 0.040 U
▪ Susceptible – Group A/S. pyogenes
▪ Resistant – All the rest species
2. SXT disk (Sulfamethoxazole – Trimethoprim)
▪ Resistant – Group A, Group B
▪ Susceptible – All the rest species
3. CAMP Test (Christie, Atkins, Munch, Petersen)
▪ Synergistic Beta hemolytic reaction between
o Staph aureus and S. agalactiae (Group B)
▪ CAMP Factor
o Diffusible, protein like compound produced by Group B
o Arrow head hemolytic reaction = S. agalactiae (Group B)
4. Hippurate Hydrolysis
➢ Hippuric acid (Sodium Hippurate) – (hippuricase)(enzyme) – to benzoic acid +
glycine
- Then add Ninhydrin (Indicator)
▪ Positive result:
o Group B (deep blue or purple)
▪ Negative result
o Other strep will give colorless reaction
5. PYR Test (Pyrrolidonyl Arylamidase)
➢ L pyrrolidonyl Beta naphthylamide – PYR – Beta naphthalinamin
➢ Indicator:
o P- dimethylaminocinnamaldehyde (D MACA)
➢ Positive result: Red
o Group A , enterococcus
➢ Negative result
o Orange
6. LAP (Leucine Aminopeptidase)
Leucine Beta naphthylamide – LAP – Beta naphthylamine
Indicator: P dimethylaminocinnamaldehyde (DMACA)
➢ Positive result: Red
▪ S. pneumonia
▪ S. pyogenes
▪ Group B Enterococcus
7. Bile Esculin Hydrolysis
▪ 40% Bile
▪ Ability to grow and hydrolyze Esculin
➢ Positive result: Blackening of Colony
▪ Group D (Enterococcus and Non enterococcus)
Negative: No blackening of the agar)
▪ (Viridans Streptococcus)
8. Salt Tolerance
▪ 6.5% NaCl
▪ To differentiate Group D enterococcus and non enterococcus
➢ Positive result: Growth
o Group D enterococcus
➢ Negative result: No Growth
o Group D Non enterococcus
9. Optochin Susceptibility (Taxo P)
- Ethylhydrocupreine HCL
- Used to differentiate Streptococcus pneumoniae from other α-hemolytic
streptococci
-
Cut off
▪ > 14 mm (6mm Disk)
▪ > 16 mm (10mm Disk)
➢ Susceptible: S pneumoniae (more than cut off >)
➢ Resistant: All other Strep (less than cut off <)
10. Bile Solubility Test
➢ 2% bile (Na Deoxycholate or Na Taurocholate)
➢ Bile – turbid solution
➢ Test for the solubility to bile salts
➢ Positive result: Colony disintegrates
o S. pneumonia – has autocatalytic enzyme activity
➢ Negative result: Intact Colonies
o Other α-hemolytic strep
Species Lancefield Hemolysis Common Terms
S. pyogenes A β Group A Strep
S. agalactiae B β Group B Strep
S. dysgalactiae C β Group C Strep
S. equi
S. bovis group D α,γ Group D – non enterococcus
E. faecalis, E. faecium D α,β,γ Group D – enterococcus
S. pneumoniae -------- α Pneumococcus
Anginosus, mutans, mitis A,C,F,G,N β, α, γ Viridans Strep