The Staphylococci

HIBA SALEEM
LECTURER
 The Staphylococci
• The staphylococci are gram-positive spherical cells, usually arranged in grapelike
irregular clusters.

• The genus Staphylococcus has at least 45 species.

The four most frequently encountered species of clinical importance are
 Staphylococcus aureus
 Staphylococcus epidermidis,
 Staphylococcus lugdunensis
 Staphylococcus saprophyticus.

• S. aureus is a major pathogen for humans.

• Almost every person will have some type of S.aureus infection during a lifetime,
ranging in severity from food poisoning or minor skin infections to severe life-
threatening infections.
• The coagulase-negative staphylococci (CoNS) are normal human microbiota and
sometimes cause infection, often associated with implanted devices, shunts, and
intravascular catheters, especially in very young, old, and immunocompromised
patients.
The Staphylococci
Morphology and Identification

1.Organism

2.Culture

3.Growth Characteristics

4.Antimicrobial Resistance
 1. Typical Organism
The characteristics of Staphylococci are;
 Spherical cells about 1 μm in diameter
 Arranged in irregular clusters
(Single cocci, pairs, tetrads, and chains)
 Young cocci stain strongly gram positive
 On aging, many cells become gram
negative.
 Staphylococci are non-motile.
 Non spore-former
 Aerobic
 2. Culture
 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.
 No pigment is produced anaerobically or in broth.
 3. Growth Characteristics
 Catalase Positive- which differentiates them from the streptococci.
 Slowly ferment many Carbohydrates, producing lactic acid but not gas.
 Staphylococci are relatively resistant to drying, heat (they withstand 50°C
for 30 minutes), and 10% sodium chloride.
 4. Antimicrobial Resistance
 Staphylococci are variably susceptible to many antimicrobial drugs.
Resistance is caused by several mechanisms:

 β-Lactamase production is common, is under plasmid control, and makes

the organisms resistant to many penicillin's (penicillin G, ampicillin,
ticarcillin, piperacillin,

 Resistance to nafcillin (and to methicillin and oxacillin) is independent of β-

lactamase production. Resistance to nafcillin is encoded and regulated by a
sequence of genes found in a region of the chromosome called “The
Staphylococcal Cassette Chromosome mec (SCCmec)”. This cassette encode
a low-affinity penicillin-binding protein (PBP2a) that is responsible for the
resistance. (MRSA)-Methicillin Resistant Staphylococcus aureus.
Staphylococcal Cassette Chromosome mec
(SCCmec)
 Antimicrobial Resistance
 Vancomycin-intermediate S.aureus (VISA)- Isolated in Japan.

• Mechanism of Resistance
The mechanism of resistance is associated with increased cell wall
synthesis and alterations in the cell wall

The isolates contained the Vancomycin resistance gene vanA likely

derived from enterococci and the nafcillin resistance gene mecA.
Pathogenesis of Staphylococci
 Pathogenesis

 PMN: Polymorphonuclear neutrophils- the most abundant innate

immune cells in the body and act as the first defense against infections
 Langerhans cells (LC) are tissue-resident macrophages of the skin
 Pathogenesis
1. Infection (0-2 Hours)
Tha S. aureus bacteria adhere and penetrate in the epidermal part of the skin. After
penetration the bacteria starts of colonize and an inflammatory response is generated.

2. Inflammatory Response (2- 24 Hours)

In inflammatory response the Langerhan’s cells of the skin starts the process of phagocytosis
with the help of first line of defense mechanism (PMN- Polymorphonuclear neutrophils).
The bacteria escapes from the process of phagoctosis and hence resulting in the damage of
PMN which lead to the necrosis of PMN’s.

3. Abscess Formation (2-6 Days)

After inflammatory response, abscess formation started which contains necrotic PMNs,
Viable PMNs, bacteria and mononuclear phagocytes. These all cells are then covered by a
fibrous capsule which forms an abscess.

4. Mature Abscess (6- 14 Days)

The abscess is then matured containing the mass of viable PMNs, necrotic lysed PMN (pus
formation), extracellular DNA, this whole is now known as a fibrous capsule.
Diseases/ Infections
Diagnostic Laboratory Tests
Diagnostic Laboratory Tests
1.Specimen
2.Smear
3.Culture
4.Catalase Test
5.Coagulase Test
6.Susceptibility Testing
7.Serologic and Typing Tests
 1. Specimen
 Surface swab pus or aspirate from an abscess.
 Blood
 Endonasotracheal aspirate
 Expectorated sputum
 Spinal fluid for culture, depending on the localization of the process

2. Smear
• Staphylococci appear as gram-positive cocci in clusters in smears of pus or
sputum.
 2. Culture
 Specimens planted on blood agar plates give rise to typical colonies in 18
hours at 37°C, but hemolysis and pigment production may not occur until
several days later and are optimal at room temperature.
 Mannitol salt agar (MSA) or commercially available chromogenic media are
used to screen for nasal carriers of S.aureus
 4. Catalase Test
This test is used to detect the presence of catalase 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
 5. Coagulase Test
Citrated rabbit (or human) plasma is mixed with an equal volume of broth culture or
growth from colonies on agar and incubated at 37°C.
A tube of plasma mixed with sterile broth is included as a control.
If clots form in 1–4 hours, the test result is positive.
 6. Susceptibility Testing
Clinical laboratories adopt methods recommended by the Clinical and Laboratory
Standards Institute (CLSI) or European Committee on Antimicrobial
Susceptibility Testing (EUCAST) for the performance of susceptibility testing of
staphylococci. Resistant to Penicillin G
 Broth Microdilution
Resistant to Nafcillin- Cefoxitin
 Disk diffusion susceptibility

6. Serologic Tests
 Pulsed-field gel electrophoresis (PFGE)
 Sequence Typing
Pulse Field Gel Electrophoresis (PFGE)
 Treatment
 Flucloxacillin
 Gentamycin
 Fusidic acid
 Rifampicin
 Erythromycin
 Vancomycin
 Cefotaxime
 Methicillin
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