Lecture 7 Gram Positive Pathogen (Staphylococci) - 1

Muhammad Hilal Jan
GRAM POSITIVE COCCI Mphil Microbiology

BS MLT
GRAM POSITIVE COCCI
Two medically important genera
• Staphylococcus and Streptococcus
Both are non motile and non spore forming
Distinguished by two main criteria

• Staphylococcus appears in grape like cluster while
Streptococcus appear in chains
• Biochemically, Staphylococci produces Catalase while
Streptococci do not
STAPHYLOCOCCUS
GENERAL CHARACTERISTICS
Gram-positive cocci, found in irregular grapelike clusters
Like all medically important cocci, they are nonflagellate,

nonmotile, and non-spore-forming
Staphylococci grow best aerobically but are facultatively

anaerobic and produce catalase unlike streptococci
CONT…
More than one dozen species of staphylococci colonize

humans; of these, three are of major medical
importance:
• S. aureus
• S. epidermidis
• S. saprophyticus.
The most important species from the viewpoint of human
medicine is S. aureus. Other species are common in the
skin flora but produce lower grade disease.
S. AUREUS VS OTHER STAPHYLOCOCCI
Production of coagulase by S. aureus
Production of carotenoid pigment (Staphyloxanthin) which

impart golden color to the colonies of S. aureus
Virulence of S. epidermidis is significantly less than S.

aureus
S. aureus ferments mannitol and hemolyze red blood cells

where as others do not. (S. aureus produces hemolysins)
FEATURES OF HUMAN STAPHYLOCOCCI
STAPH AUREUS
90% of Staph aureus contain plasmid that encode beta lactamase (enzyme that
degrade many but not all penicillins.
Some strains are even resistant to methicillin (MRSA) and nafcillin (NRSA) as they
produces altered penicillin binding proteins (PBP). (mecA genes encodes these altered
PBPs)
Some strains have with intermediate and full resistance to vancomycin (VISA and
VRSA respectively) have also been detected.
Genes responsible for vancomycin resistance are located in a transposons on plasmid

that encode enzyme which substitute D-lactate for D-alanine in peptidoglycan
CELL WALL COMPONENTS AND ANTIGENS
The cell wall of S. aureus consists of a typical Gram-positive peptidoglycan interspersed
with molecules of a ribitol-teichoic acid
• In most strains, the peptidoglycan of the cell wall is overlaid with

surface proteins; one protein, protein A, is unique in that it binds the Fc
Protein A portion of IgG molecules, It probably contributes to the virulence of S.
aureus by interfering with opsonization. Protein A is also used for
diagnostic purposes.
Teichoic acid • It mediate adherence of staphylococci to mucosal cells.

(polymers of
ribitol • Lipoteichoic acid also play role in Induction of septic shock by
Phosphate) inducing cytokines like IL-1 and TNF
CELL WALL COMPONENTS AND ANTIGENS
• Important virulence factor

• 11 serotypes based on antigenicity but type 5 and
Polysaccharide type 8 cause 85% of infections
Capsule • Microcapusle: small amount of polysaccharide
capsule
• Capsule is poorly immunogenic
Peptidoglycan of S. aureus has endotoxin-like property and can

cause clinical finding of septic shock yet not posses endotoxins.
TRANSMISSION
Humans are reservoir Hand contact is

for staphylococci. important mode of Sneezing
Nose and main site transmission
of colonization.
Contaminated Food
Surgical wounds • Custard, Pastries, Potato
salad, Canned meats
PATHOGENESIS
S. aureus causes disease both by producing toxins and by inducing pyogenic
inflammation.
The typical lesion of S. aureus infection is an abscess. Abscesses undergo central
necrosis and usually drain to the outside (e.g., furuncles and boils), but organisms may
disseminate via the bloodstream as well.
Foreign bodies, such as sutures and intravenous catheters, are important predisposing
factors to infection by S. aureus.
PATHOGENESIS
Several important toxins and enzymes are

produced by S. aureus. The three clinically
important exotoxins are enterotoxin, toxic
shock syndrome toxin, and exfoliatin
TOXINS AND BIOLOGICALLY ACTIVE
EXTRACELLULAR ENZYMES
α-Toxin α -Toxin is a protein secreted by almost all strains of S.
aureus but not by coagulase negative staphylococci. It
lyses cytoplasmic membranes by direct insertion into the
lipid bilayer to form transmembrane pores. The resultant
egress of vital molecules leads to cell death
P-V is a pore-forming toxin that kills cells, especially white

leukocidin blood cells, by damaging cell membranes. The two subunits
of the toxin assemble in the cell membrane to form a pore
through which cell contents leak out.
An individual strain may produce one or more toxins but less than
10% of S. aureus strains produce any PTSAg. As superantigens
they are strongly mitogenic for T cells and do not require
proteolytic processing prior to binding with class II major
histocompatibility complex (MHC) molecules on antigen-
presenting cells
Pyrogenic Toxin
Superantigens This stimulates both T cells and macrophages to release massive
amounts of cytokines, particularly interleukin-1, interleukin-2 and
tumor necrosis factor
It causes “scalded skin” syndrome in young children in
which intercellular splitting of the epidermis between the
stratum spinosum and stratum granulosum, presumably by
disruption of intercellular junctions
Exfoliatin
The enzymes include coagulase, fibrinolysin, hyaluronidase,
proteases, nucleases, and lipases. Coagulase, by clotting
plasma, serves to wall off the infected site, thereby
retarding the migration of neutrophils into the site.
Staphylokinase is a fibrinolysin that can lyse thrombi.
Staphylococcal Superantigens Toxin
The superantigen (SAgs) are family of secreted proteins that are able to stimulate systemic
effects.
There are more than 15 described StaphSAgs, the most important is Toxic Shock Syndrome
Toxin (TSST-1) and enterotoxin.
Less than 10% of S. aureus strains produce any StaphSAg.
Superantigens are highly mitogenic for T Cells and do not require proteolytic processing
before binding with class II MHC molecules, which results in massive cytokine release.
Compared to a normal antigen-induced T-cell response where 0.0001-0.001% of the body’s
T-cells are activated, these SAgs are capable of activating up to 20% of the body’s T-cells.
TSST causes toxic shock, especially in
tampon-using menstruating women or in
individuals with wound infections.
Toxic shock Toxic shock also occurs in patients with

syndrome toxin nasal packing used to stop bleeding
(TSST) from the nose.
TSST is produced locally by S. aureus in
the vagina, nose, or other infected site.
The toxin enters the bloodstream, causing a toxemia. Blood
cultures typically do not grow S. aureus.
TSST is a superantigen and causes toxic shock by stimulating the

Toxic shock release of large amounts of IL-1, IL-2, and TNF.
syndrome
toxin (TSST) Approximately 5% to 25% of isolates of S. Aureus carry the
gene for TSST. Toxic shock occurs in people who do not have
antibody against TSST.
Signs and symptoms including high fever, accompanied by
low blood pressure, malaise and confusion, which can rapidly
progress to stupor, coma, and multiple organ failure.
Enterotoxin causes food poisoning characterized by prominent vomiting and watery,
nonbloody diarrhea.
It acts as a superantigen within the gastrointestinal tract to stimulate the release of

large amounts of IL-1 and IL-2 from macrophages and helper T cells, respectively.
Enterotoxin The prominent vomiting appears to be caused by cytokines released from the
lymphoid cells, which stimulate the enteric nervous system to activate the vomiting
center in the brain.
Enterotoxin is fairly heat-resistant and is therefore usually not inactivated by brief
cooking. It is resistant to stomach acid and to enzymes in the stomach and jejunum.
There are six immunologic types of enterotoxin, types A–F.

CLINICAL MANIFESTATION
The important clinical manifestation
caused by S. aureus can be divided into
two groups
Pyogenic (Pus Toxin

producing) Mediated
PYOGENIC INFECTION
Skin Infections
• Impetigo, Furuncles, Carbuncles, eyelids infection, cellulitis, folliculitis,
hidradenitis, lymphangitis etc
Septicemia
Endocarditis
Osteomyelitis
Postsugical wound Infection
Pneumonia
Conjunctivitis
Abscesses
Impetigo
Furuncles Hidradenitis
Eyelid Infection
Conjunctivitis (Blepharitis)
Lymphangitis Hordeolum Folliculitis
Osteomylitis
Abscess
Endocarditis
TOXIN MEDIATED DISEASES
Food Toxic Shock Scalded Skin

poisoning Syndrome Syndrome
LABORATORY DIAGNOSIS
Gram stained smear and culture. Yellow or golden colonies on blood agar, colonies
are often B-hemolytic
S. aureus is catalase positive, coagulase positive and DNAase positive.
Serological tests are not useful
TREATMENT
Penecillin G for sensitive isolates, B-lactamase resistant penicillins such as nafcillin for
resistant isolates, Vancomycin for isolates resistant to nafcillin. About 85-90% are
resistant to penicillin G.
Prevention
 Cefazolin is used to prevent surgical wound infection. Hand washing reduces spread. No vaccines are
available
STAPHYLOCOCCUS EPIDERMIDIS
Characteristics
 Gram positive cocci in clusture, white colonies on blood agar, catalase positive and coagulase negative
and Novobiocin Sensitive
Habitat and transmission

 Normal flora of the human skin and mucous membrane. It is probably the patients own strains that
cause infection but transmission from person to person via hands may occur
Diseases
 Endocarditis on prosthetic heart valves, prosthetic hip infection, intravascular catheter infection,
cerebrospinal fluid shunt infection and neonatal sepsis
Pathogenesis
• Glycocalyx producing strains adhere well to foreign
bodies such as prosthetic implants and catheters.
• It is a low virulence organism that causes disease
primarily in immunocompromised patients and in those
with implants. It is a major cause of nosocomial
infections.
• Unlike S. Aureus no exotoxins have been identified.
Gram stained smear and culture. Whitish non
Laboratory hemolytic colonies on blood agar. It is catalase
diagnosis positive, coagulase negative and novobiocin sensitive.
Vancomycin plus either rifampin or an aminoglycoside.

It produces B-lactamases and is resistant to many
Treatment antibiotics
Prevention There is no vaccine for S. epidermidis

STAPHYLOCOCCUS SAPROPHYTICUS
Gram positive cocci in clusters
White colonies on blood agar. Catalase positive, coagulase negative

and novobiocin resistance
Normal flora of mucosa of genital track in young women
Causes UTI, particularly in sexually active young women, most women

with this infection have had sexual intercourse within the previous 24
hours.

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Muhammad Hilal Jan

GRAM POSITIVE COCCI Mphil Microbiology

Both are non motile and non spore forming

Distinguished by two main criteria

Gram-positive cocci, found in irregular grapelike clusters

Like all medically important cocci, they are nonflagellate,

Staphylococci grow best aerobically but are facultatively

More than one dozen species of staphylococci colonize

Production of carotenoid pigment (Staphyloxanthin) which

Virulence of S. epidermidis is significantly less than S.

S. aureus ferments mannitol and hemolyze red blood cells

Genes responsible for vancomycin resistance are located in a transposons on plasmid

• In most strains, the peptidoglycan of the cell wall is overlaid with

Teichoic acid • It mediate adherence of staphylococci to mucosal cells.

• Important virulence factor

Peptidoglycan of S. aureus has endotoxin-like property and can

Humans are reservoir Hand contact is

Several important toxins and enzymes are

P-V is a pore-forming toxin that kills cells, especially white

Toxic shock Toxic shock also occurs in patients with

TSST is a superantigen and causes toxic shock by stimulating the

It acts as a superantigen within the gastrointestinal tract to stimulate the release of

There are six immunologic types of enterotoxin, types A–F.

Pyogenic (Pus Toxin

Postsugical wound Infection

Food Toxic Shock Scalded Skin

Habitat and transmission

Vancomycin plus either rifampin or an aminoglycoside.

Prevention There is no vaccine for S. epidermidis

White colonies on blood agar. Catalase positive, coagulase negative

Normal flora of mucosa of genital track in young women

Causes UTI, particularly in sexually active young women, most women

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