STREPTOCOCCUS

Dr. Kritika
Genus Characteristics

 The streptococci and enterococci are Gram-positive cocci

that occur singly, in pairs, or in chains of varying lengths.
Each coccus is less than 2 µm in diameter and can appear
as spherical or ovoid.
 The streptococci are fastidious and require the addition
of blood or serum to media for growth
HABITAT

 Streptococci are world wide in distribution.

 Most of the streptococci of Veterinary interest live as commensals in the mucosa
of the upper respiratory and lower urogenital tracts.
 They do not survive for long away from the animal hosts.
CULTURAL CHARACTERISTICS

 They are facultative anaerobes, catalase negative, oxidase-negative, and non-spore forming
and non-motile with exception of some of the enterococci.
 On blood agar after incubation for 24hrs small, circular, semi-transparent colonies with an
area of clear haemolysis are produced.
 Virulent strains from fresh isolates produce matt (finely granular) colony and avirulent
strains form glossy colonies. Strains producing capsules form mucoid colonies.
 In glucose or serum broth, growth occurs as a granular turbidity with a powdery deposit.
MORPHOLOGY

 In Edwards‘s medium (selective media) it produces dewdrop like black colonies.

Edwards medium contain blood agar, crystal violet and aesculin (differentiate among
different species of streptococci which do or do not hydrolyse aesculin).
 S. pneumoniae is alpha haemolytic, produces mucoid or flat colonies with smooth borders
and a central concavity after 48-72hrs on blood agar (draughts man colonies).
 Ability to grow in 0.1 % tellurite broth is characteristic of S. faecalis. Majority of
streptococcal species do not grow on Mac Conkey agar except Enterococcus faecalis.
 Capsulation is not a regular feature of streptococci but some strains of S.pyogenes and
some group C strains have capsules composed of hyaluronic acid.
 while polysaccharide capsules are encountered in members of group B and D.
 Protoplasts (L-forms) may be induced by penicillin or phage associated lysine and may be
propagated on hyper tonic media.
BIOCHEMICAL PROPERTIES

 Streptococci are catalase and oxidase negative .

 Ferment several sugars producing acid but no gas.
 They ferment sorbitol, trehalose, lactose, maltose, dextrin, and mannitol.
 Gelatin not liquified.
 Nitrates not reduced.
 Indole is negative.
 They are not soluble in 10% bile unlike pneumococci
CLASSIFICATION

 Several systems of classification have been employed.

 Based on growth characteristics, type of haemolysis and biochemical activities they
can be divided into 6 principal categories

Group Examples
1. Pyogenic Streptococci S. pneumoniae, S.pyogenes, S.equi, S. dysgalactiae
2. Oral Streptococci S.salivarius
3. Enterococci S.faecalis, S.avium, S. gallinarum
4. Lactic acid Streptococci S.lactis
5. Anaerobic Streptococci S. monbillorum
6. Other Streptococci S.bovis, S.uberis, S.equi subsp zooepidemicus
The type of haemolysis produced by a streptococcal species can be variable.
The main types of haemolysis are:
 (β) Beta-haemolysis: a clear zone of haemolysis around the colony
 (α) Alpha-haemolysis: a zone of greening or of partial haemolysis
 (γ) Gamma-haemolysis: no haemolysis
 Most of the pathogenic streptococci fall into the beta group and are called as the
haemolytic streptococci.
 ά - streptococci are generally commensals in the throat. Because of the distinctive green
color, they produce; they are called as greening or viridans streptococci. Eg. S.pneumoniae
 Gamma or non-haemolytic streptococci produces no change in the medium.
 The gamma streptococci includes the faecal streptococci ( S. faecalis) and related species.
They are called the enterococcus or indifferent streptococci.
Lancefield Groups
 The serological Lancefield grouping scheme is based on group-
specific carbohydrate cell wall antigens, with groups from A to H
and K to V. Some isolates are not groupable. The methods for
Lancefield grouping include:
1. Conventional method: the C-substance (antigen) is extracted
either by autoclaving or by acid extraction (hydrochloric acid). A
ring precipitation test is conducted by layering the extracted
antigen over known antisera that can be obtained commercially
for some Lancefield Groups.
2. Latex agglutination test: This method is the most commonly used
and kits are available commercially for identifying Lancefield
Groups A, B, C, D, F and G
ANTIGENECITY

 The hyaluronic acid capsule of S. pyogenes inhibits phagocyotosis.

 The cell wall is composed of -
-an outer layer of fimbria containing proteins and lipoteichoic acid,
-a middle layer of group specific carbohydrate and
-an inner layer of peptidoglycan.
 The Cell wall polysaccharide has been shown to have a toxic effect on connective tissue in experimental animals.
 The peptidoglycan is responsible for cell wall rigidity. Several protein antigens have been identified in the cell wall (M,
T and R).
 Among these the M protein acts as a virulence factor by inhibiting phagocytosis.
 Hair-like pili project through the capsule of group A streptococci.
 The pili consist partly of M protein and are covered with lipoteichoic acid which is important in the attachment of
streptococci to epithelial cells.
TOXINS AND VIRULENCE FACTORS

1. Extra cellular toxins - Hemolysins

Streptolysins O and S are produced by groups A, C and G.
1. Streptolysin O
 It is so called because it is oxygen labile.
 It is an antigenic protein which is heat labile.
 It is lethal on I/V inj. into animals and has a specific cardiotoxic activity. It is a general
cytotoxin.
 Streptolysin O is antigenic and anti-streptolysin regularly appears in sera following
Streptococcal infection.
 Estimation of this antibody (ASO) titre is a standard serological procedure for the
retrospective diagnosis of infection with S.pyogenes.
2. Streptolysin S
 It is an oxygen stable haemolysin and so is responsible for the beta haemolysis seen around
streptococcal colonies on the surface of blood agar plates.
 It is called Streptolysin S since it is soluble in serum. Addition of serum to broth increased the
yield of haemolysin.
 It is protein but not antigenic. It has been shown experimentally to be nephrotoxic.
3. Erythrogenic toxin (Streptococcal pyogenic exotoxins/ Dick toxin)
 Four erythrogenic toxins are known and most strains of S. pyogenes produce one or more. They
are pyogenic and enhance susceptibility to lethal shock by endotoxin.
 The toxin is thermostable and antigenic. The intradermal injection in rats leads to development
of erythema.
 This reaction is called as ―Dick test or Schultz-charlton reaction and it is useful for diagnosis of
scarlet fever.
2. Enzymes
1. Streptokinase (Fibrinolysin): Filtrates of streptococci gp, A, E & G produces fibrinolysin. This
toxin promotes the lysis of fibrin clots by activating a plasminogen.
 Fibrinolysin plays a biological role in streptococcal infections by breaking down the fibrin
barrier around the lesions and facilitating the spread of infection.
2. Deoxyribonucleases (Streptodornase): These cause depolymerization of DNA, pyogenic
exudates contains large amount of DNA, derived from the nuclei of necrotic cells.
 Streptodornase helps to liquefy the thick pus and may be responsible for the thin serous
character of streptococcal exudates.
 Four antigenically distinct streptodornase, A, B, C & D have been recognized.
3. Hyaluronidase: This enzyme breaks down the hyaluronic acid of the tissues. This might favour
the spread of infection along intercellular spaces.
 Streptococci possess a hyaluronic acid capsule and also elaborate a hyaluronidase- a
seemingly self-destructive process.
 This is produced by group A, C, G and B Streptococci.
4. Proteinase: This is another instance of an apparently self-destructive enzyme, since it is
capable of breaking down the M protein, streptokinase and hyaluroindase.
 The enzyme is, however, produced only under special conditions such as an acidic pH (5.5 –
6.5). Such conditions may be produced by tissue destruction, as in abscesses.
 Most strains of S. pyogenes form proteinase.
5. Neuraminidase: This activity is detected in streptococci groups A, B, C, G and L. This enzyme is
a virulence factor for pathogens surviving on mucosal surface.
PATHOGENESIS
DIAGNOSIS

It involves clinical, microscopical and bacteriological examination.

1. Clinical examination
 Palpation of the udder and supramammary lymph nodes will be helpful in distinguishing the
chronic and insidious form of mastitis produced by S.agalactiae and S.uberis.
 In contrast S. dysgalactiae and S. zooepidemicus causes sudden onset of acute inflammation
of one quarter only with an acute systemic disturbance followed by joint infections and
lameness.
2. Microscopical examination
 When long chains of organisms are detected in milk samples from chronic mastitis, it is
caused by S. agalactiae.
3. Based on type of haemolysis on blood agar
4. Bacteriological examination

 When 0.1 ml of secretions inoculated on Edward‘s

medium (blood agar, crystal violet and aesculin) S.
agalactiae produces bluish-grey colonies and S. uberis
produces dark color colonies.
1. CAMP test (Christie, Atkins, Munch and
Peterson, 1944)
 This test is based on the observation that
ruminant red blood cells lysed by the beta
toxin of staphylococci at 37°C are completely
lysed in the presence of S. agalactiae (group
B).
 Differentiation between the pneumococcus
and S.viridans organisms can be achieved by
bile solubility and the optochin test.
2. Bile solubility test
 Autolysis of pneumococcal cultures takes place within 15 minutes
at 37°C in the presence of 10 per cent sodium deoxycholate.
 These substances have no effect on S. viridans organisms.
3. The optochin test
 The majority of pneumococcal strains are sensitive to optochin
(Ethyl hydrocuprein hydrochloride).
 This test consists of placing a small circular piece of filter paper,
impregnated with 1:4000 aqueous solution of optochin, in the
center of a blood agar plate after inoculating the test cultures in
streaks across the full width of the medium.
 The growth of pneumococcal strains will be inhibited to a distance
of some 5 mm from the circumference of the filter paper.
5. Molecular characterization
Thank You