Gram positive cocci

Genus: Staphylococcus spp

Genus: Staphylococcus spp

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The genus Staphylococcus contains about 30 species. Only some of them are
important as human pathogens :

1- Staphylococcus aureus (Staphylococcus pyogenes)

Is responsible for most Staphylococcal infections and aureus causes a variety of

suppurative (pus-forming) infections. Nasal carriage occurs in 20 – 50% in
human.

2- Staphylococcus epidermidis (Staphylococcus albus)

Usually present as normal flora of human skin and mucous membrane (non
pathogenic) but may cause infection in immune –compromised patient if
accidentally introduced catheterization.

3- Staphylococcus saprophyticus. Free living non- pathogens but may produce

U.T.I

Morphology and Identification

A. Typical Organisms
Staphylococci are spherical cells about 1 µm in diameter arranged in irregular
clusters . Single cocci, pairs, tetrads, and chains are also seen in liquid cultures.
Young cocci stain strongly Gram-positive; on aging, many cells become
Gramnegative. Staphylococci are nonmotile and do not form spores. They are
found free living in the environment and form regular packets of four (tetrads)
or eight cocci. Their colonies can be yellow, red, or orange. Micrococci are
rarely associated with disease.

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B. Culture
Staphylococci grow readily on most bacteriologic media under aerobic or
microaerophilic conditions. They grow most rapidly at 37°C but form pigment
best at room temperature (20–25°C). Colonies on solid media are round,
smooth, raised, and glistening. S. aureus usually forms gray to deep golden
yellow colonies. S. epidermidis colonies usually are gray to white on primary
isolation; many colonies develop pigment only upon prolonged incubation.

C. Growth Characteristics

The staphylococci produce catalase, which differentiates them from the

streptococci. Pathogenic staphylococci produce many extracellular substances,
which are discussed below. Staphylococci are relatively resistant to drying, heat
(they withstand 50°C for 30 minutes).

Staphylococci extracellular enzymes & toxins

The main pathogenesis is by invasion + multiplication + enzymes & toxins
production
The bacterial enzymes are under plasmids or chromosomal control and they
are:
1- Catalase: that convert H2O2 to H2O and O2 in Staph but not in Strep

2- Coagulase & clumping factor:

3- Hyalorunidase, staphylokinase that produce fibrinolysis, proteinase, lipase.
The toxins are:
1- α-toxins (hemolysin) act upon leucocytes cell wall
2- - β-toxins act upon RBCs act upon leucocytes wall

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3- γ-toxins (hemolysin) act upon leucocytes wall
4- δ-toxins act upon human epithelial cells
5- Exfoliative toxins (A+B) they are epidermolytic toxins, cause skin
desquamation (scalded skin), type A is under chromosomal control and
it heat stable while type B is under plasmid control and it heat labile.
6- Toxic shock syndrome toxin (TSST-1) it bind to major
histocompatability complex class II (MHC-II) molecules and this will
lead to T-lymphocytes stimulation.
7- Enterotoxins (emetic effect) and it types are (A-E, G-J, K-R, and U, V);
it preformed as Staph grow in carbohydrate or protein diets and the
personnel ingest the already produced toxins not the microbe itself.

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Diagnostic Laboratory Tests

The specimens are pus swab, blood, tracheal or abscess aspirates, urine, CSF

Direct Smear Examination

1-Gram stained smears

Gram-positive cocci pairs

and clusters

Numerous polymorphonuclear cells (PMNs

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2- Characteristics Culture

- Cultural onto 5% blood agar plate (BAP), Mannitol salt plate as S aureus ferment
it to gain yellow colonies.
Colony morphology

Smooth white to yellow, creamy

S. aureus may produce hemolysis on blood agar

3-Catalase test

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 This test is used to detect the presence of cytochrome oxidase enzymes. A
drop of 3% hydrogen peroxide solution is placed on a slide, and a small
amount of the bacterial growth is placed in the solution. The formation of
bubbles (the release of oxygen) indicates a positive test result.

Principle: tests for enzyme catalase

2H2O2 2 H2O + O2

Drop H2O2 onto smear

Bubbling = POS (Most bacteria, O2 generated)

No bubbling = NEG (Streptococci and other lactic acid bacteria, no O2

generated)

4-Hemolysis in BAP

5-Coagulase test

This test used to differentiate between S. aureus & other Staphylococcus species
(CNS). which is carried out in 2 ways (slide); we use particles covered with
fibrinogen and IgG antibodies that bind coagulase to gain clumping within 20
seconds if the IgG is present in low concentration we turn onto tube method as we
mix equal volumes from bacterial broth and citrated plasma then incubate it for 1-4
hours at 37 ºC to gain the clot in positive reaction (Coagulase positive Staph)
(CPS); S intermedius usually give positive test although it is not regarded as
human pathogen. ((Most of the CPS is pathogenic to human))

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Staphylococci infections

Local infection or abscess with painful inflammatory reaction that goes to central
suppuration and heals as soon the pus is drained. The fibrin and immune cells
around the lesion try to prevent infection spread, so must prevent its manipulation
or trauma rupture.

Staphylococci occur also directly through wound contamination (surgical or

traumatic) as meningitis after skull fracture or osteomyelitis after open fracture
surgery. If the bacteria transmitted by blood stream we get bacteremia, pulmonary,
osteomyelitis, meningitis, arthritis, etc.

In food poisoning the incubation period is short (1-8 hours); with vomiting and
diarrhea & NO FEVER.

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