Streptococcus and
other related cocci
Classification:
Phylum : Firmicutes
Class : Bacilli
Order : Lactobacilliales
Family : Streptococcacea
Genus : Streptococcus
Species : agalactiae
dysagalactiae
equi Sub sp. Zooepidemicus
uberis
equi sub sp. equi
canis
suis
pyogenes
pneumoniae
History
Rivolta (1873) described chain forming organisms in pus
from a case of strangles in horses.
In 1878-79, Pasteur recognized this organism as a pus-
forming agent.
In 1903, Hugo Schottmuller introduced blood to
differentiate various types of hemolysis.
In 1928, Rebecca Lancefield reported a serological
method of grouping streptococci.
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.
Classification systems:
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.
Aerobic and facultative anaerobic streptococci
are classified as per their haemolytic properties
Brown (1919) established this method by employing meat
infusion peptone agar with 5% horse blood. He recognized
three types of reactions.
o alpha - haemolytic streptococci: They produce a greenish
discolouration with partial haemolysis around the colonies.
o The zone of lysis is small (1or2 mm wide), within which the
unlysed erythrocytes are seen.
o These ά - streptococci are generally commensals in the
throat. Because of the distinctive green color, they produce;
they are called as greening or viridans streptococci.
[Link]
o Beta haemolytic streptococci produces a sharply
defined, clear, colourless zone of haemolysis, 2 or 4mm
wide, within which the red cells are completely lysed.
o Most of the pathogenic streptococci fall into the beta
group and are called as the haemolytic streptococci.
o Gamma or non-haemolytic streptococci produces no
change in the medium.
o The gamma streptococci includes the faecal streptococci
( S. faecalis) and related species. They are called the
enterococcus or indifferent streptococci.
Lancefield classification:
• by Rebecca lancefield (1933) was
• based on the nature of a carbohydrate (C) antigen on the cell
wall of beta haemolytic streptococci
•. They are known as Lancefield groups, 19 of which have been
identified so far and named by the capital letters A-U (without I
and J) .
Haemolytic streptococci of group A are
known as [Link] . These may be further
subdivided into types based on the protein
(M,T and R) antigens present on the cell wall.
The M protein is acid and heat labile and the T protein
is acid labile and trypsin resistant.
Some of the Lancefield groups may be further
subdivided by means of the agglutination test and
designated by Arabic numbers-Griffith typing
Cultural characters:
It is an aerobe and facultative anaerobe, growing best at a
temperature of 37°C. They grow best in media enriched with blood,
serum and fermentable carbohydrates.
On blood agar after incubation for 24hrs small, circular,
semitransparent 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. No pellicle is formed.
In Edwards’s medium (selective media) it produces dewdrop like
black colonies.
Edwards medium containing 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.
Bio-chemical 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
Resistance:
It is a delicate organism easily destroyed by heat (54° C for 30
minutes).
It can survive in dust for several weeks, if protected from sunlight.
It is rapidly inactivated by antiseptics.
It is more resistant to crystal violet and susceptible to
sulphonamides and other antibiotics.
Sensitivity to bacitracin is employed as a convenient method for
differentiating S. pyogenes from other hemolytic streptococci.
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).
They are responsible for type specificity in [Link]. 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 & virulence factors
Hemolysins
Streptolysins O and S are produced by groups A, C and G.
Streptolysin O
It is so called because it is oxygen labile. It is an antigenic protein
and is active in the reduced form.
On blood agar, its activity is seen only in pour plates and not in
surface cultures. It is also heat labile.
It is lethal on I/V inj. into animals and has a specific cardiotoxic
activity. It is a general cytotoxin.
Red cells of all animal species except mouse are lysed.
Streptolysin O is antigenic and antistreptolysin 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
[Link].
Toxins & virulence factors
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. 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.
Toxins & virulence factors
Enzymes
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.
Deoxyribonucleases (Streptodornase): These cause depolymerisation
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.
Toxins & virulence factors
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.
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.
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. In addition to this M types of [Link] produce
NADase and other many strains also produce esterase, amylase and N-
acetyl glucosaminidase.
Pathogenecity:
Cattle : Bovine mastitis
It is caused by S. agalactiae (group B), S. dysgalactiae (grp C), S.
equi subsp. zooepidemicus (grp. C), S. uberis (grp C, D, E, P, V).
Mastitis arises from the multiplication of streptococci in the teat
sinus and extends into the ducts.
It causes parenchymatous mastitis, which is characterized by
progressively chronic condition resulting in fibrosis.
In acute stages milk is composed of purulent exudate, dead tissue
cells, coagulated milk protein and bacteria.
Peptostreptococcus indolicus is an anaerobic streptococcus, which
is responsible for summer mastitis in cattle in association with
Arcanobacterium pyogenes.
Horse
S. equi and S. equisimilis are the main causes of strangles in
young horses.
It is characterized by a catarrhal discharge, with inflamation of the
nasal mucous membranes, followed by swelling of pharyngeal
Lymph Nodes in which abscesses develop.
The infection spreads through lymph channels. It also causes
metritis and cervicitis in horses.
Purpura haemorrhagica, considered to be an immune mediated
disease, occur in horses 1 to 3 weeks after illness.
Bastard strangles – in which abscesses developed in many
internal organs. It is a very serious complication.
Chicken
S. gallinarum causes typical acute septicemia with
peritonitis in chicken.
Dogs
S. canis is considered to be the cause of acid milk in
puppies and canine tonsillitis. It is also associated
with neonatal septicaemia and toxic shock syndrome.
Pigs
S. suis causes porcine cervical lymphadenitis and
also isolated from pneumonia, septicaemia, arthritis,
endocarditis, meningitis and reproductive tract
infections.
It also causes erosive arthritis in young pigs.
Diagnosis:
Clinical examination
Palpation of the udder and supramammary
lymphnodes will be helpful in distinguishing the chronic
and insidious form of mastitis produced by [Link]
and [Link].
In contrast [Link] and S. zooepidemicus
causes sudden onset of acute inflammation of one
quarter only with an acute systemic disturbance
followed by joint infections and lameness.
Microscopical examination
When long chains of organisms are detected in milk
samples from chronic mastitis, it is caused by S.
agalactiae.
Based on type of haemolysis on blood agar
Bacteriological examination
When 0.1 ml of secretions inoculated on Edward’s
medium (blood agar, crystal violet and aesculin)
[Link] produces bluish-grey colonies and [Link]
produces dark color colonies.
CAMP test (Christie, Atkins, Munch and Peterson, 1944)
o This test is based on the observation that ruminant red blood
cells lysed by the beta toxin of staphylococci at 37ᵒC are
completely lysed in the presence of S. agalactiae (group B).
Staphylococcus
Streptococcus agalactiae aureus
Optochin sensitivity test
o The majority of pneumococcal strains are sensitive to optochin
(Ethyl hydrocuprein hydrochloride). Whereas [Link] organisms
are not.
o 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.
o The growth of pneumococcal strains will be inhibited to a
distance of some 5 mm from the circumference of the filter paper.
Control & prevention
In streptococcal infections in animals, including mastitis,
the most satisfactory method of control is antibiotic therapy
using penicillin preparations to other substances because
streptococci develop resistance to penicillin comparatively
infrequently.
Vaccines are of very limited value for the immunization of
animals against streptococcal infections, with the possible
exception of equine strangles.
[Link] (group. A) is the most usual cause of septic
sore throat and scarlet fever in humans.