MICROCOCCEAE

EPIDEMIOLOGY
 Staphylococcus aureus

▪ present among the indigenous flora of the skin, eye, upper respiratory
tract, gastrointestinal tract, urethra, and, infrequently, vagina.
▪ leading causes of bacteremia in hospitalized patients,
▪ Most common cause of spinal epidural abscess and suppurative
intracranial phlebitis, and may be recovered from brain abscesses,
typically following trauma.
▪ Most common cause of septic arthritis in pre-pubertal children
 Staphylococcus aureus

TOXINS
▪ exotoxin TSST-1 is responsible for toxic shock syndrome
▪ illness is most common in women 15–25 years of age who use
tampoons during menstruation
▪ enterotoxins A–E are responsible for staphylococcal food poisoning.
▪ exfoliative toxins—epidermolytic toxins A and B—cause skin erythema
and separation, as seen in scalded skin syndrome.
▪ CA-MRSA - Panton-Valentine leukocidin toxin (PVL)
 Protein A: This protein has sites that bind the Fc
portion of IgG. This may protect the organism from
opsonization and phagocytosis.
Coagulase: This enzyme can lead to fibrin formation
around the bacteria, protecting it from phagocytosis.
Penicillinase: This is a secreted form of beta-lactamase.
It disrupts the beta-lactam portion of the penicillin
molecule, thereby inactivating the antibiotic

Hemolysins (4 types): Alpha, beta, gamma, and

delta. They destroy red blood cells, neutrophils,
macrophages, and platelets.
Leukocidins: They destroy leukocytes
Proteins to Tunnel Through Tissue
1) Hyaluronidase ("Spreading Factor"): This
protein breaks down proteoglycans in
connective tissue.
2) Staphylokinase: This protein lyses
formed fibrin clots (like streptokinase).
3) Lipase: This enzyme degrades fats and
oils, which often accumulate on the surface
of our body. This degradation facilitates
Staphylococcus aureus' colonization of
sebaceous glands.
4) Protease: destroys tissue proteins.
 OTHER STAPHYLOCOCCI

S. epidermidis
▪ most frequently involved in association with foreign bodies,
especially implanted prosthetic valves, joints, and shunts
S. saprophyticus
▪ an important cause of bacteriuria, particularly among sexually
active young women
 LABORATORY DIAGNOSIS

MICROSCOPY
 LABORATORY DIAGNOSIS

▪ 5% sheep blood and chocolate agars.

▪ They also grow well in broth-blood culture systems and common
nutrient broths, such as thioglycollate, dextrose broth, and brain-heart
infusion
▪ Phenylethyl alcohol (PEA) or Columbia colistin-nalidixic acid (CNA)
agars may be used to eliminate contamination by gram-negative
organisms in heavily contaminated specimens such as feces.
 LABORATORY DIAGNOSIS

▪ Mannitol salt agar - contains a high concentration of salt (10%), the

sugar mannitol, and phenol red as the pH indicator.
▪ S. aureus ferments mannitol and produces a yellow halo on this
media as a result of acid production altering the pH
 LABORATORY DIAGNOSIS

Incubation Conditions and Duration

▪ Visible growth on 5% sheep blood and chocolate agars incubated at
35° C in carbon dioxide (CO2) or ambient air usually occurs within 24
hours of inoculation.
▪ Mannitol salt agar and other selective media may require incubation
for at least 48 to 72 hours before growth is detected.
LABORATORY DIAGNOSIS
LABORATORY DIAGNOSIS
 Coagulase Test

▪ S. aureus produces two forms of coagulase:

▪ bound
▪ free.
▪ Bound coagulase, or “clumping factor,” is bound to the bacterial cell
wall and reacts directly with fibrinogen.
▪ The presence of bound coagulase correlates with free coagulase
▪ Free coagulase an extracellular protein enzyme that causes the
formation of a clot when S. aureus colonies are incubated with plasma.
Coagulase Test
 Coagulase Test

▪ Staphylococcus lugdunensis and Staphylococcus schleiferi are two

other staphylococci that may give a positive result with this slide
coagulase test.
▪ For the tube coagulase test, several colonies are transferred into a
tube containing plasma that is incubated at 35° C for 4 hours and
then is examined for clot formation.
 Coagulase Test

▪ test should be examined after 4 hours because most isolates of S.

aureus produce a clot within this interval, but some strains produce a
fibrinolysin that can lyse the clot, thus producing a false-negative
reaction if the test is observed only after 24 hours.
▪ Staphylococcus intermedius and Staphylococcus hyicus will also be
positive with the tube coagulase test.
 Antimicrobial Susceptibility

▪ 90% of staphylococci are resistant to penicillin

▪ due to an inducible plasmid-encoded β-lactamase
▪ mediated by the mecA gene, which encodes an altered penicillin-
binding protein, PBP-2a.
▪ 1-μg oxacillin disk has been used traditionally for disk diffusion testing
 Antimicrobial Susceptibility

▪ 30-μg cefoxitin disk test is a better indicator than oxacillin

▪ isolates with zone sizes ≥22 mm can be reported as susceptible (S),
and those with zone sizes ≤21 mm can be reported as oxacillin
resistant
▪ Oxacillin in cation supplemented Mueller-Hinton broth containing 2%
NaCl – microdilution testing
▪ To screen isolates of S. aureus for oxacillin resistance, Mueller-Hinton
agar supplemented with 4% NaCl and containing 6 μg/mL of oxacillin is
spot inoculated with a cotton swab, and plates are incubated for 24
hours at 35° C.
Antimicrobial Susceptibility