Staphylococcus

Staphylococceae family
 Cocci (spheres)
 Grapelike clusters
 3 planes
 1 micrometer diameter
 Nonmotile
 Halotoerant
 Catalase (+)
 Oxidase (-)
 +/- fermentation
 Staphylococcus species
 61 species
 Important
 S. aureus
 S. epidermidis
 S. saprophyticus
 S. pseudointermedius
 S. lugdunensis
 Others
 S. haemolyticus
 S. capitus
 S. hominis
 S. xylosis
 S. warneri
Location
 Ubiquitous
 Normal flora
 Skin
 Mouth
 Nose
 Throat
 Predisposing factors
 Immunosuppression
 Concurrent disease
 AB resistance
Epidemiology
 Normal microbiotica
 Direct transmission
 Portal of Entry
 Hair follicle
 Break in skin
 Respiratory
 Complications
 Bacteremia
 Septecemia
 Abscess
 Pathogenesis
 Immune evasion
 Slime layer biofilm
(microcapsule)
 Evades phagocytosis
 Protein A on cell wall
 Binds to IgG @ Fc
Blocks opsonization
 Adhesin proteins
 Binds to
fibrin/fibrinogen
 Binds to ECM
laminin
fibronectin
collagen
Enzymes
 Coagulase
 Catalase
 Hemolysins
 Panton–Valentine
Leukocidin
 Hyaluronidase
 Lipase
 Beta lactamase
 Staphylokinase
(plasminogen activator)
 Proteases
 DNase
 FAME (fatty acid
modifying enzyme)
 Staphylococcus Toxins
 Cytolytic Toxins
 Alpha = β hemolysis
 Beta = sphingomyelinase
 Delta
 Gamma
 Leukocidin = hemolysis
 Leukotoxin = WBC destruction
 Exfoliative toxin
 ETA and ETB
 Protease & esterase activity
 Toxic Shock toxins
 Enterotoxins (A-E)
Superantigens
 Enterotoxin
 Food poisoning, V/D
 TSS if B or C antigenic type
 TSST-1: desquamation; TSS
Virulence Factors Summary
Virulence Factors: Extracellular enzymes

 Cytolytic Toxins
Alpha hemolysin: lyses rbcs, damages plts,
causes severe tissue damage
ß hemolysin: acts on sphingomyelin in the
plasma membrane of rbcs
Virulence Factors: Extracellular enzymes

 Hyaluronidase: Hydrolyzes hyaluronic acid in

connective tissue allowing spread of infection

 Staphylokinase: fibrinolysin which allows spread of

infection

 Coagulase: virulence marker

 Lipase: allows colonization

Virulence Factors: Extracellular enzymes
 Beta-lactamase or Penicillinase: confers
resistance

 DNase: degrades DNA

 Protein A: in cell wall, it binds to Fc part of IgG

toblock phagocytosis
 Staph aureus

 Distinguishing Features
 • Small, yellow Staphylococcus aureuscolonies on blood
agar
 • β-hemolytic
 • Coagulase positive (all other Staphylococcus species
are negative)
 • Ferments mannitol on mannitol salt agar
 Staph aureus
 Location
 Mucous membranes
 Moist areas
 Diseases
 Noninvasive
 Food poisoning
 Enterotoxin (V/D)
 Cutaneous
 Scalded skin
 Impetigo
 Folliculitis
 Boils, styes, furunculosis
 Systemic
 Soft tissue infections
Staphylococcal disease summary
 Reservoir
 • Normal flora
 –– Nasal mucosa
(25% of population are  Transmission
carriers)  • Hands
 • Sneezing
 –– Skin
 • Surgical wounds
 • Contaminated food
 –– Custard pastries
 –– Potato salad
 –– Canned meats
 Predisposing Factors for
Infection
 • Surgery/wounds
 • Foreign body (tampons, surgical packing, sutures)
 • Severe neutropenia (<500/μL)
 • Intravenous drug abuse
 • Chronic granulomatous disease
 • Cystic fibrosis
 Pathogenesis
 Protein A binds Fc component of IgG, inhibits phagocytosis

 Enterotoxins: fast acting, heat stable

 Toxic shock syndrome toxin-1 (TSST-1): superantigen

 Coagulase: converts fibrinogen to fibrin clot

 Cytolytic toxin (α toxin): pore-forming toxin, Panton-Valentine leukocidin

(PVL), forms pores in infected cells and is acquired by bacteriophage;
associated with increased virulence, MRSA strains

 Exfoliatins: skin-exfoliating toxins (involved in scalded skin syndrome

[SSS]) and bullous impetigo
 Yellow colonies of S. aureus on a blood agar plate, note regions of clearing around colonies caused by lysis of
red cells in the agar (beta hemolysis)

 Laboratory Diagnosis:
Direct Smear Examination

Microscopic Examination
1. Gram-positive cocci
2. pairs and clusters
3. Numerous polymorphonuclear cells (PMNs)
Laboratory Diagnosis:
Cultural Characteristics

 Colony morphology
 Smooth, butyrous,
white to yellow,
creamy
 Grow well in 18-24
hours
 S. aureus may produce
hemolysis on blood
agar
S. aureus
Identification Tests: Catalase

 Principle: tests for enzyme catalase

 Drop H2O2 onto smear

 Bubbling = POS (Staph)

 No bubbling = Streptococci
Identification Tests: Coagulase Test

 Cell-bound “clumping factor” Two methods

converts fibrinogen to fibrin
which precipitates on cell Slide test: screens
causing agglutination for “clumping factor”
 Extracellular enzyme “free
coagulase”
Tube test
Novobiocin Susceptibility Test

 Test to differentiate coagulase-negative staphylococci

from S.saprophyticus from urine samples
 S. saprophyticus is resistant (top)
 Treatment
Gastroenteritis is self-limiting.

Nafcillin/oxacillin are drugs of choice because of

widespread penicillinase-producing stains.

 Mupirocin for topical treatment.

For methicillin-resistant Staphylococcus aureus (MRSA):

vancomycin

For vancomycin-resistant Staphylococcus aureus (VRSA)

or vancomycin- intermediate S. aureus (VISA):
quinupristin/dalfopristin
MRSA
 Hosptial associated
 Community assoc.
AMA/CDC poster
VRSA

vanA operon from enterococcus changes cell wall tripeptide sequence

Opportunistic Staph pathogens

 S. capitis
 S. epidermidis
 S. saprophyticus
 S. xylosis
 S. epidermidis  S. saprophyticus
 • Coagulase (−); gram (+) cocci  • Coagulase (−), gram (+) cocci
 • Novobiocin sensitive  • Novobiocin resistant
 • Infections of catheters/shunts

Novobiocin sensitivity testing A. Novobiocin-

resistant (S. saprophyticus) 
B. Novobiocin Sensitive (S. epidermidis)
S. epidermidis
 Location
 Normal skin flora
 opportunistic pathogen
 Skin/wound infections
 Endocarditis
 UTI
 Exposure
 Direct contact
 Newborns
 Elderly
 Fomites
 Catheters
 Shunts
 IV needles
 Prosthetics
S. saprophyticus
 Pathogenesis
 Fimbria
 Adhesion proteins
 Autolysins
 Diseases
 UTI/cystitis
 Peritonitis
 Enopthalmitis
 Endocaritis
 Septic arthritis
Rapid Staph test
Immunology

 Hyperimmune serum
 Capsular proteins
 Surface adhesins
 Vaccination
 At risk population
 ESRD
 Dialysis patients
 Premature infants
 StaphVAX
 Capsular proteins
 Exotoxin A (Ps. Aeurginosa)