Gram-positive Cocci

Staphylococci
‘BUNCH OF GRAPES
 Gram-positive Cocci
• Coccus ‘ Spherical shape’.

• Two medically important genera of gram-positive cocci: Staphylococcus

and Streptococcus and are responsible for a wide variety of clinical
diseases.

• Staphylococci appear in grapelike clusters, whereas streptococci are in

chains.

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 STAPHYLOCOCCUS

 The most important species associated with human diseases:

1) Staphylococcus aureus (S. aureus).

2) Staphylococcus epidermidis (S. epidermidis).
3) Staphylococcus saprophyticus (S. saprophyticus).

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4
 PROPERTIES

 Characteristics:

• Spherical gram-positive cocci arranged in grapelike

clusters.
• Catalase-Test: positive (degrades H2O2 into O2 and H2O).
• Coagulase-Test: positive (coagulase activates pro-
thrombin to form thrombin, causing blood to clot).
• β-hemolytic (completely hemolyze RBCs on an agar
plate).
Gram stain of S. aureus cells which
• Facultative anaerobic. typically occur in clusters

• Non-spore forming.

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 PROPERTIES

 Habitat and Transmission:

 Main habitat:
• Exist as a normal flora on skin and mucosal surfaces of human.
• S. aureus is also found in the vagina of approximately 5% of women.
• All over hospitals.
 Transmission:
• Staphylococcus aureus is always spread by physical contact, not through the air.
• Staphylococcus aureus can spread directly from the pus of an infection like boils or
abscesses through skin-to-skin contact.
• Through indirect contact with contaminated objects such as sheets, towels, sport equipment.
• Presence of foreign body (suture, catheter).
• Unnecessary antibiotic use for years.
• Staphylococcus aureus can survive on dry surfaces for up to several months. 
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 VIRULENCE FACTORS

 S. aureus has numerous powerful weapons stick out of the

microcapsule:

1) Protein A: This protein has sites that bind immunoglobulin IgG, preventing
opsonization and phagocytosis.

2) Coagulase: Allows fibrin formation around organism, protecting it from phagocytosis.

3) Catalase. Conversion of hydrogen peroxide to water and oxygen.

4) Hemolysins: β-hemolysis (complete lysis of red blood cells) on an agar plate which
appears as a yellow golden pigment on sheep blood agar. Hemolysins destroy RBCs,
macrophages, and platelets.
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 VIRULENCE FACTORS

5) Leukocidins: Destroy leukocytes (WBCs).

6) Penicillinase (β-lactamase): This enzyme produced by S. aureus that provide

resistance to beta-lactam antibiotics like penicillins. It disrupt the beta-lactam
portion of the penicillin molecule, thereby inactivating the antibiotic by
breaking the antibiotics' structure.

7) Hyaluronidase: Breaks down connective tissue facilitating S. aureus spread in

tissue.

8) lipases: Degrades fats and oil on the surface of our body. This degradation
facilitates S. aureus’ colonization of glands.
9) Protease: Destroys tissue proteins.
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 CLINICAL DISEASES

 Staphylococcus aureus causes a wide range of human diseases which can be

separated into two groups:
A. Diseases Caused By Exotoxin Release.
B. Disease Resulting From Direct Organ Invasion.

C. Diseases Caused By Exotoxin Release:

1) Enterotoxin (Gastroenteritis, food poisoning):
• Staphylococci can grow in food produce an exotoxin (e.g., ham, pastries, potato salad,
and ice cream).
• The victim will then eat the food containing the pre-formed toxin.
• Rapid onset of disease within 4 hours.
• Food poisoning characterized by vomiting, watery diarrhea, abdominal pain, and
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occasionally fever. Lasts 12 to 24 hours.
 CLINICAL DISEASES

2) Toxic Shock Syndrome Toxin-1 (TSST-1):

• TSST-1 causes toxic shock, especially in tampon-using menstruating women.
• TSST is produced locally by S. aureus in the vagina. The toxin release into the blood
stream, causing a toxemia causes high fever, nausea and vomiting, watery diarrhea,
and rash.
• This diseases may involve three or more organs: liver, kidney. Muscle, blood,
gastrointestinal, or central nervous system (CNS).

3) ‘Staphylococcal Scalded Skin Syndrome (SSSS) or ‘Ritter’s disease’.

• A Staphylococcus aureus strain, which produces exfoliative toxin A and B.
• It is characterized by fever, rash, hair and nails can be lost. Recovery usually occurs
within 7-10 days.
• Usually affects neonates and young children.
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 CLINICAL DISEASES

TSST-1 Scalded skin syndrome (SSS)

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 CLINICAL DISEASES

B. Disease Resulting From Direct Organ Invasion:

1) Local infection:

• Skin infection are very common.

A. Impetigo:
• usually occurs on the face, especially around the
mouth.
• Small vesicles lead to pustules.
B. Cellulitis:
• This is a deeper infection of the cells.
• The tissue becomes hot, red, shiny and swollen.
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 CLINICAL DISEASES
C) Local Abscesses, Furuncles, and Carbuncles:

• Abscesses is a collection of pus. Infection of hair follicle

produces pus with a red rim. This infection can penetrate
deep into the subcutaneous tissue to become a furuncle. These
may produce painful lesions communicating under the skin
called carbuncles.

D) Wound infections:

• Any skin wound can be infected with S. aureus after surgical

procedure.
• Their clinical signs are: increasing local pain, redness,
swelling.
• If the wound does not heal, antibiotic therapy should be used
against S. aureus. 13
Wound infection Abscesses

Carbuncles Furuncle

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 CLINICAL DISEASES

2)Systemic diseases:

A.Pneumonia:
• S. aureus pneumonia usually follows a viral influenza (flu) upper respiratory illness, with
onset of fever, chills, rapid destruction of lung, resulting in cavitaions (holes in the lung),
and pus.
• In many hospitals, S. aureus is the most common cause of nosocomial pneumonia.

B.Meningitis, Cerebritis, Brain Abscess:

• These patients can present with high fever, stiff neck, headache, coma, and neurologic signs.

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CLINICAL DISEASES
Staphylococcus aureus pneumonia

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 CLINICAL DISEASES

C. Osteomyelitis: Osteomyelitis
• This is a bone infection that usually
occurs in children <12 years of age.

• The infection spreads to the bone,

presenting locally with warm,
swollen tissue over the bone, fever,
and shakes.

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 CLINICAL DISEASES

D. Arthritis:

• Invasion of the synovial membrane by Staphylococcus

aureus.
• Patients complain of painful red swollen joints with
decreased range of motion.
• S. aureus is the most common pathogen causing this
disease in children and adults over the age of 50.
• Without prompt treatment, many patients will
permanently lose the function of the joints.
• Antimicrobial therapy is required.

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 CLINICAL DISEASES

E. Endocarditis:
• This is a destructive infection of the heart valves.
• A sudden onset of high fever, chills, and myalgia.
• Intravenous drug (IV) users develop endocarditis.

F. Blood and catheter infection:

• Staphylococcus aureus can migrate from the skin
and colonize central venous catheters resulting in
bacteremia, sepsis, and endocarditis.

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 MRSA FACT SHEET
• MRSA is Methicillin-Resistant
Staphylococcus aureus.
• More than 90% of S. aureus contain
plasmid that encode β-lactamase
(penicillinase) which provide resistance
to antibiotics such as penicillinase-
resistant penicillins (methicillin,
nafcillin, oxacillin, etc.).
• MRSA is a dangerous type of S. aureus
causes skin and other infections.
• MRSA tend to develop in the hospital,
where broad spectrum antibiotics are
used.
• MRSA is spread by the hand contact with
another infected patient, sharing personal
items (towels, razors, etc.), touching
surface or items contaminated with
MRSA. 20
 LABORATORY DIAGNOSIS

• Serious staphylococcal infections such as infection of the bloodstream,

pneumonia, and endocarditis require culturing of samples of blood or
infected fluids.

• Infected wound (either a small biopsy of skin or pus taken with a swab) or of
the blood must be obtained to grow the bacteria in the microbiology
laboratory. Once the Staph is growing, the organism is tested to determine
which antibiotics will be effective for treating the infection.

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 LABORATORY DIAGNOSIS

1) Gram Stain: S. aureus lie in grape-like clusters as seen on gram stain.

Staphylococcus aureus is a typical Gram-positive bacterium forming

irregular clusters of cocci. 22
 LABORATORY DIAGNOSIS
2) Culture:
A. β-hemolytic.
B. Produces a golden yellow pigment on sheep blood agar.

Staphylococcus aureus cultivated on Columbia agar with

5% defibrinated sheep blood. Cultivation 24 hours in an
aerobic atmosphere, 37°C. Colonies are surroundend by a
wide zone of beta-hemolysis. 23
 LABORATORY DIAGNOSIS
3) Metabolic:
A. Catalase Test: positive.
B. Coagulase Test: positive (Slide coagulase test, tube coagulase test).

Catalase test-positive (blowing Slide coagulase-positive Tube coagulase-positive,

oxygen bubbles) clot plasma into gel and
do not flow when inverted
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 PREVENTION  

 Vaccine is NOT currently available against Staphylococcus aureus.

1) Practice good hygiene, with careful hand washing.

2) clean cuts, scrapes, and wounds.
3) Use disposable gloves, especially in hospitals and nursing homes, to avoid skin-to-
skin contact.
4) Avoid skin contact with an infected person.
5) Use ethanol as a topical sanitizer against MRSA.
6) Use proper food handling and preparation to prevent food poisoning.

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PREVENTION

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 PROPERTIES

 Characteristics:

• Gram positive cocci in clusters.

• Catalase test-positive.
• Coagulase test-negative.

 Habitat and Transmission:

• Habitat: normal flora of the human skin and mucosal membranes.

• Transmission: via hands.

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 VIRULENCE FACTORS
• Polysaccharide Capsule: adhere to a variety of prosthetic devices such as
prosthetic joints, prosthetic heart valves, and peritoneal dialysis catheters.
• Highly resistant to antibiotics.

Clinical diseases
• S. epidermidis enters the blood stream causing bacteremia.

• Infections of intravenous catheters sites and prosthetics materials such as

prosthetic heart valves (endocarditis), prosthetic joints (arthritis or
osteomyelitis), sepsis in neonates, and large wounds.

• It is the major cause of hospital acquired infections. It is very likely to

contaminate patient-care equipment and environmental surfaces.

• Unlike, S. aureus, no toxins have been identified.

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 LABORATORY DIAGNOSIS
1) Gram stain:
• Gram positive cocci in clusters.
2) Culture:
• Whitish, non-hemolytic colonies on blood agar

Golden S.aureus and white S.epidermidis colony

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 LABORATORY DIAGNOSIS

3) Metabolic:
• Catalase test-positive.
• Coagulase test-negative.
4) Antibiotic test:
• Sensitive to novobiocin antibiotic

Catalase-test positive Novobiocin-sensitive 31

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 PROPERTIES
 Characteristics:

• Gram positive cocci in clusters.

• Catalase test-positive.
• Coagulase test-negative.

CLINICAL DISEASES

• Urinary tract infection in women.

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 LABORATORY DIAGNOSIS
1) Gram stain:
• Gram positive cocci in clusters.

2) Culture:
• Gamma-hemolytic.

3) Metabolic:
• Catalase test-positive.
• Coagulase test-negative.

Novobiocin-resistant
4) Antibiotic test:
• Resistant to novobiocin antibiotic

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Gram Positive Cocci

GPC

Catalase-test

H2O2 into O2 and H2O

Staphylococci

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 Coagulase-test

S. aureus S. epidermidis or S. saprophyticus

Methicillin Novobiocin

R S R

MRSA S. epidermidis S. saprophyticus

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