STREPTOCOCCUS and ENTEROCOCCUS (Gram positive cocci)

STREPTOCOCCACEAE

- Gram positive organisms

can be a normal flora

or a pathogenic bacterium

 Enterococci formerly known as Group D Streptococci

 Lactococci formerly known as Group N
 Under the Streptococcaceae
o Streptococci
o Aerococcus
o Lactococcus
o Leuconostoc
o Pediococcus

GENERAL CHARACTERISTIC of Streptoccci

 Gram positive cocci in pairs or chain

 Catalase (-)
o To organism it does not have catalase or an enzyme which is the catalase
o Use by the bacteria to convert oxygen into a water or non-free radical type of ion
 Facultative anaerobes
o Can live with or with oxygen
 Capnophilic (can stimulate the growth of Carbon dioxide (CO2))
o 5-10% carbon dioxide needed, stimulates the growth and other parts of bacteria
 Colonies are small and transparent
 Non-motile

CLASSIFICATION

-Help us in diagnostic of streptoccoci

 Bergey’s Classification
o Based on the temperature requirements
 Lancefield
-common
o Based on the C substrate of the cell wall (polysaccharides)-commonly capsule or
carbohydrate substrate
o Classification is developed by Rebecca Lancefield.
 Brown
o Based on the hemolysis
BERGEY’S CLASSIFICATION

PYROGENIC VIRIDANS LACTIC ENTEROCOCCUS

Temperature 37*C 45 & 37 10 &37 10, 45, 37
Hemolytic Pattern Common beta Alpha and non- Non-hemolytic Non-hemolytic
hemolytic hemolytic
Example S. Pyogenes S. Mutans S. Lactic E. faecalis

 Hemolytic / hemolysis of organism- can help us to understand, what organism or what would be
the possible organism

BROWN’S CLASSIFICATION HEMOLYSIS

-hemolytic pattern is commonly seen in blood culture media, cannot be seen in chocolate agar, because
hemoglobin in chocolate agar is already lyse

 Presumptive identification, particularly of streptococci and enterococci.

o a-Hemolysis is partial lysing of erythrocytes in BAP around and under the colony that
results in a green discoloration of the medium
o B-Hemolysis is complete clearing of erythrocytes in BAP around or under the colonies
because of the complete lysis of RBCs. - rbc is total lyse
o Nonhemolytic, no lysing of hemoglobin. –light will not past through
o Alpha Prime small area of intact RBCs around colony surrounded by a wider zone of
complete hemolysis.

alpha hemolysis- partial; S. pneumonia

beta hemolysis- complete; S. pyogenes

gamma hemolysis- none; E. faecalis

Streptococcus

 Beta hemolytic (clear)

o Pyogenes
 Group A, bacitracin sensitive
o Agalactiae
 Group B, bacitracin resistant
 y-hemolytic
o Enterococcus
 E. faecalis
 E. faecium
 a-hemolytic (green)
o pneumonia
 optochin sensitive
 bile soluble
 capsule => quelling +
o viridans
 mutans
 sanguis
 optochin resistant
 not bile soluble
 no capsule

LANCEFIELD CLASSIFICATION

 Based on the extraction of C carbohydrate from the streptococcal cell wall by

placing diluted acid and heat for 10 minutes
 For the identification of Beta hemolytic species
o Streptococcus agalactiae (Group B)
o Streptococcus pyogenes (Group A)

Beta hemolysis Alpha, beta hemolysis, Non hemolytic

PYROGENIC NONPYROGENIC
Group A Group B Group C Group D
Streptococcus Streptococcus Streptococcus Enterococcus faecalis
pyogenes agalactiae dysagalactiae Enterococcus faecium
Streptococcus equi Streptococcus bovis

STREPTOCOCCUS PYOGENES

 Lancefield A
  With different virulence factor:
o M protein- plays a role in adherence and resist phagocytosis
o Lipotechoic Acid, Protein F & Fibronectin Binding Protein- for the attachment of
streptococcto oral
o Hyaluronic Acid Capsule- prevent opsonization, phagocytosis by neutrophil or
macrophages.
o Streptolysin O
 Hemolysis when incubated anaerobically best to exhibit when stabbing the BAP
 Measured by Anti Streptolysin O (ASO test)
o Streptolysin S
 Hemolysis when incubate aerobically
 Oxygen stable
o Streptokinase
 Convert plasminogen --- plasmin
 Lyse clot
-Clotting help the body to enclosed a bacterium

o Hyaluronidase
 AKA spreading factor
 Enzyme that solubilize the ground substance of mammalian connective tissue
 Spread infections
 Exotoxin- Stimulate T cell proliferation

Clinical Infection:

 Strep throat/pharyngitis- (tonsillitis)

 Impetigo - (Group A) necrotizing fasciitis; flesh eating bacteria syndrome
  Erysipelas- dermal layer skin infectious; painful swelling reddish spot
 Cellulitis- redness and accumulation of fluid (pamumula at pamamaga)
 Scarlet fever- (strawberry color tongue) exotoxin; susceptibility test Dick test (show of redness
of the skin)
 Necrotizing fasciitis
 Streptococcal toxic shock- SPSS; condition whole organ system shut down and lead to death;
due to toxin
 Rheumatic Fever
o Damage heart valves
 Acute glomerulonephritis
o Occur after pharyngitis
o Complement, inflammatory response cause damage to the glomeruli

LABORATORY DIAGNOSIS

 Specimen: Throat swab

 Culture Media
o SBA in CO2 increase growth
o SBA w/ sulfamethoxazole or colistin or Polymyxin B.
 S. pyogenes at SBA: small, transparent and smooth with defined Beta hemolysis
 Bacitracin susceptible
 SXT (sulfamethoxazole) Resistant- antibiotic test
 PYR Hydrolysis Positive
 Immunologic test: ASO, Anti-Dnase, Antistreptokinase, Antihyaluronidase

STREPTOCOCCUS AGALACTIAE

 Group B (acid stable polysaccharide at cell wall)

 9 capsular polysaccharide serotype
 Microbiota of female genital tract and lower GIT
 Contains residue of sialic acid
 Virulence factor:
o Capsule
o Hemolysin- for hemolytic pattern (beta hemolytic)
o CAMP - unique virulence factor, known as Christie–Atkins–Munch-Peterson; help us to
identify if its group b streptococci or group a streptococci
o Neuraminidase
o Dnase
o Hyaluronidase

CLINICAL INFECTIONS
  Mastitis
 Disease on Newborn
o Early stage: Pneumonia & sepsis
o Late stage: Meningitis & sepsis- have a normal flora of vagina
 Health woman become ill after childbirth, abortion
 Elderly with serious condition.

LABORATORY DIAGNOSIS

 SBA: Grayish, white mucoid colonies in Beta hemolysis

 Specimen: vagina/rectal swab
 Todd Hewitt w/ Colistin & Nalidixic acid
 (+) CAMP & Hippurate test
 Agglutination test
 Resistant for SXT and Bacitracin
 Strep B Carrot Broth
o (+) Betahemol

Streptococcus pyogenes & Streptococcus agalactiae Identification

 LEUCINE AMINOPEPTIDASE TEST (LAP)

 Presumptive identification of Catalase Negative Gram + Cocci
 Detection of Leucine Aminopeptidase
 Leucinebeta-naphthylamide + leucine aminopeptidase+ cinnamaldehyde --- beta-
nephthylamine (red color)
 (+) Development of red color
 (-) No color change or slightly yellow

BACITRACIN SUSCEPTABILITY TEST

 Presumptive identification of Streptococcus pyogenes in Throat swab

 Inoculate at blood agar with SMZ and bacitracin disk.
 Susceptible: Streptococcus pyogenes
 Resistant: Streptococcus agalactiae

HIPPURATE HYDROLYSIS

 To differentiate Streptococcus agalactiae and other Beta Hemolytic Streptococci

 Reagent: Ninhydrin, Hippurate Disk
 Hippurate+ H2O---- Benzoic Acid+
 Glycine (+): Purple (Streptococcus agalactiae)
  (-): Colorless or Yellow (Streptococcus pygones)

CAMP TEST

 Christie, Atkins, Munch-Petersen

 Use beta lysin producing strain of Staphylococcus aureus
 Synergistic hemolysis between group B
Streptococcus and beta-hemolytic
Staphylococcus aureus
 (+) Enhanced hemolysis in arrowhead patter
e.g Streptococcus agalactiae
 (-) No arrowhead shown

PYRROLIDONYL-ALPHA NAPHTHYLAMIDE HYDROLYSIS

 Presumptive of Beta hemolytic

Streptococci and non-hemolytic D.

 (+): Red color

 Streptococcus pyogenes
 Enterococcus
 Aerococcus
 Gemella