FAMILY

MICROCOCCACEAE
-JOSE ANGELO ANGELES BUNAG, RMT, MSMT
 GENERA

Staphylococcus
Micrococcus
Planococcus
Stomatococcus
STAPHYLOCOCCUS
STAPHYLOCOCCUS
MICROCOCCUS
TYPES OF HEMOLYSIS
ALPHA PRIME

Colony is surrounded with

an alpha zone of
hemolysis and outer Beta
zone of hemolysis
 GROWTH ON LOEFFLER’S SERUM
SLANT

S. aureus Golden Yellow

S. citreus Lemon Yellow

S. albus Porcelein White

 GROWTH ON MANNITOL SALT
AGAR

INHIBITOR 7.5 NaCl

CHO Mannitol

PH INDICATOR Phenol red

 CATALASE TEST

RGT 3% H2O2

POSITIVE RESULT Effervescence

 TESTS TO DIFFERENTIATE
STAPHYLOCOCCUS FROM
MICROCOCCUS
TESTS STAPHYLOCOCCUS MICROCOCCUS

AEROBIC GROWTH + +

ANAEROBIC GROWTH + -

LYSOSTAPHIN SUSCEPTIBLE RESISTANT

BACITRACIN RESISTANT SUSCEPTIBLE

MODIFIED OXIDASE - +

GLUCOSE UTILIZATION FERMENTOR OXIDIZER

 CHO OXIDATION FERMENTATION
TEST
OPEN TUBE CLOSED TUBE

FERMENTOR YELLOW YELLOW

OXIDIZER YELLOW GREEN

 COAGULASE

SLIDE COAGULASE

TUBE COAGULASE
 SLIDE COAGULASE

Cell bound
DETECTS coagulase|Clumping factor

RGT Rabbit’s Plasma

POSITIVE CLOT or CLUMPING

 TUBE COAGULASE

DETECTS Free Coagulase

RGT 0.5 ml Rabbit’s Plasma

POSITIVE Gel like clot

 DIFFERENTIATION AMONG
COAGULASE + STAPHYLOCOCCI
ORGANISM TUBE VOGES- PYR
COAGULASE PROSKAUER
S. Aureus + + -
subspecies
aureus
S. intermedius V - +
S. hyicus V - -
S. scleiferi + + +
subspecies
coagulans
 DNASE - TEST

DNA medium with

MEDIUM
methyl green

POSITIVE RESULT Effervescence

 NOVOBIOCIN

• Associated with
S. epidermidis
bacterial
endocarditis
following the
insertion of artifical
heart valves
 NOVOBIOCIN

S. saprophyticus
• Important cause of
UTI in young
women
 THREE MAIN SPECIES OF CLINICAL
IMPORTANCE

Staphylococcus aureus

Staphylococcus epidermidis

Staphylococcus saprophyticus
STAPHYLOCOCCUS
AUREUS
STAPHYLOCOCCUS AUREUS

• Gram +, non-motile,
0.8 to 1.0um in
diameter in irregular
grapelike clusters
• Few strains produce a
capsule/slime layer
 METABOLISM
Energy is obtained via respiratory and fermentative
pathways
Exists under conditions of both high and low
oxidation reduction potential

Catalase is produced aerobically

Wide range of sugars and other carbohydrates are

used
 CAPSULE
Protects the bacteria by inhibiting chemotaxis and
phagocytosis by polymorphonuclear leukocytes and
proliferation of mononuclear cells following mitogen
exposure

Interferes with the interaction between the

underlying teichoic acid peptidoglycan complex
and complement
 PEPTIDOGLYCAN LAYER
Elicits the production of IL-1 and opsonic antibodies by
monocytes

Chemoattractant for polymorphonuclear leukocytes

Produces a localized Shwartzman phenomenon

Increased antipeptidoglycan IgG level in infections

accompanied by bacterimic phase
 SCHWARTZMAN PHENOMENON
• Is a rare reaction of body to particular types of
toxins called endotoxins, which cause
thrombosis in the affected tissue
• A clearing of thrombosis results n a
reticuloendothelial blockade, which prevents
re-clearing of the thrombosis caused by a
repeat introduction of the toxin. That will cause
tissue necrosis
 PROTEIN A
• A group of specific antigen unique to S. aureus
• Consists of single polypeptide chain
• Has five regions:
• Four highly homologous domains – Fc binding
• Fifth, C terminal domain – bound to the cell wall
and does not bind Fc
• Binds to the Fc portion of the IgG molecules
except IgG3
 TEICHOIC ACIDS

Ribitol teichoic acid with

N-acetyl-D-glucosamine
S. aureus

S. epidermidis Glycerol teichoic acid

with glucosyl residues
 CLUMPING FACTOR

Component in the cell wall that results in the

clumping of whole staphlococci in the presence of
plasma

Protein which binds fibrinogen and differs from free

coagulase in both its mechanism of action and its
antigenic properties
 CYTOPLASMIC MEMBRANE

A complex of protein, lipids and a small amount of

carbohydrate forming an osmotic barrier for the cell
 EPIDEMIOLOGY
• Widespread in the environment
• Can be cultured from various objects and environmental
surfaces
• May be transmitted by multiple routes, including contact
with infected persons, contact with asymptomatic carriers,
airborne spread, and contact with contaminated objects
• Colonizes the skin and mucous membranes of 30 – 50% of
healthy adults and children
• Anterior nares is the most densely colonized and can persist
for years in 10 – 20% of affected people
 DETERMINANTS OF
PATHOGENICITY
SURFACE RECEPTORS
POLYSACCHARIDE
PROTEINS
EXTRACELLULAR ENZYMES
COAGULASES
LIPASES
HYALURONIDASE
STAPHYLOKINASE (FIBRINOLYSIN)
NUCLEASE
TOXINS
CYTOLYTIC TOXINS
ENTEROTOXINS
EXFOLLIATIVE TOXIN
TOXIC SHOCK SYNDROME TOXIN - 1
 SURFACE RECEPTORS
POLYSACCHARIDE

• Surface components that possess antiphagocytic activity

are advantageous to the staphylococcus in its initial
establishment in the host
• Encapsulated staphylococci are able to spread rapidly
through tissue by protecting the organisms fro the
complement mediated attack of polymorphonuclear
leukocytes
 SURFACE RECEPTORS
PROTEIN RECEPTORS

• Specific binding sites on the staphylococcal cell surface

• Provide the organism with an adhesion mechanism by
which infective foci become established
• Also binds to components of the extracellular matrix
• Laminin
• Collagen
• Fibronectin
 PROTEIN RECEPTORS
FIBRONECTIN LAMININ

• Metastasis like potential of

staphylococci to breach
• Mediates the adherence of
the normal barriers between
vital cells such as fibroblasts,
host tissues may be related
epithelial cells, and
to its ability to bind
monocytes to an injured site
specifically to basement
membrane
 EXTRACELLULAR ENZYMES
COAGULASE

• May cause the formation of fibrin layer around the

staphylococcal abscess thus localizing the infection and
protecting the organism from phagocytosis
 EXTRACELLULAR ENZYMES
LIPASES

• Required for the invasion of staphylococci into the

cutaneous and subcutaneous tissues into the formation of
superficial skin infections
 EXTRACELLULAR ENZYMES
HYALURONIDASE

• Hydrolyzes the hyaluronic acid present in the intracellular

ground substances of connective tissue
 EXTRACELLULAR ENZYMES
STAPHYLOKINASE

• A proteolytic enzyme with fibrinolytic activity

• Can dissolve clots – proenzyme plasminogen is converted
to the fibrinolytic enzyme plasmin
 EXTRACELLULAR ENZYMES
NUCLEASES

• Phosphodiesterase with both endonucleolytic and

exonucleolytic properties and can cleace either DNA or
RNA
 TOXINS
CYTOLYTIC TOXINS

Streptolysin O and S

Various toxins of Clostridium

Hemolysins and Leukocidin of S. aureus

 ALPHA TOXIN
• Cytotoxic for a variety of tissue culture cells
• Human macrophages and platelets are
damaged; monocytes resistant
• Cause injury to the circulatory system, muscle
tissue and to the renal cortex
• Contributes to the pathogenicity by producing
tissue damage after the establishment of a focus
of infection
 BETA TOXIN

• Heat labile protein – toxic for a variety of cells, including

RBC, macrophages and fibroblasts
• Catalyze the hydrolysis of membrane phospholipids in
susceptible cells
• With alpha toxin – responsible for the tissue destruction
and abscess formation characteristics of staphylococcal
diseases and the ability of S. aureus to proliferate in the
presence of vigorous inflammatory response
 DELTA TOXIN

• Detergent like properties – have damaging effects on

membrane
• High content of hydrophobic amino acids
• When localized, becomes amphipathic and strongly
surface active
• Inhibits water absorption by the ileum
• Stimulates accumulation of adenosine monophosphate
• Alters ion permeability in the guinea pig ileum
 GAMMA TOXIN

• Contains two protein components that act synergistically

both essential for hemolysis and toxicity
• Elevated specific neutralizing antibodies in human
staphylococcal bone diseases – suggestive of its role in
the disease state
 GAMMA TOXIN
LEUKOCIDIN

1. Attacks PMNs and macrophages but no other cell

types
2. Two protein components (S and F) that act
synergistically to induce cytolysis
3. Unique response of leukocyte to leukocidin –
altered permeability to cation
 GAMMA TOXIN
PYROGENIC PROTEIN TOXINS

1. Enterotoxins
2. Toxic shock syndrome toxin 1
3. Streptococcal pyrogenic exotoxin A through C
 ENTEROTOXIN

ENTEROTOXIN A

EMETIC REEPTOR SITES

ENTEROTOXIN INDUCED DIARRHEA

SUPER ANTIGENS
 TOXIC SHOCK SYNDROME TOXIN 1
• An exotoxin with pronounced and diverse
immunologic effects
1. Induction of IL-2
2. Receptor expression
3. Interleukin synthesis
4. Proliferation of human T lymphocytes
5. Stimulation of IL-1 synthesis by human
monocytes
 TOXIC SHOCK SYNDROME TOXIN 1
• EXFOLIATIVE TOXIN
• Mediates staphylococcal scalded syndrome
• Produced by bacteriophage group II strain
• Does not elicit an inflammatory response
• Does not primarily cause cell death
• Potent mitogen primarily of T cells
• A shingomyelinase different from Beta toxin
 PATHOGENESIS

Typical Organisms
staphylococcal skin penetrate a
infection sebaceous gland or
hair shaft where the
environment is
suitable for grwoth
 PATHOGENESIS

Precipitating causes
of staphylococcal 1. Third degree burns
2. Traumatic wounds
disease 3. Surgical incisions
4. Decubitis or trophic
ulcers
5. Certain viral infections
CLINICAL MANIFESTATIONS
CLINICAL MANIFESTATIONS
CLINICAL MANIFESTATIONS
 CLINICAL MANIFESTATIONS
TOXIC SHOCK SYNDROME

FOOD POISONING|GASTROENTERITIS

SCALDED SKIN SYNDROME

BULLOUS IMPETIGO

STAPHYLOCOCCAL SCARLET FEVER

STAPHYLOCOCCAL SCALDED SKIN
SYNDROME
 TREATMENT
1. Localized staphylococcal infection
a. Adequate drainage
b. Debridement
c. Antibiotics
d. Initial drug of choice – penicllinase – resistant drugs
2. Cutaneous infection
1. Oral therapy with semisynthetic penicillin
2. Nafcillin and oxacillin – not well absorbed orally
3. Erythromycin – if allergic to penicillin
 TREATMENT

1. Serious systemic staphylococcal disease

a. Nafcillin or oxacillin – parenteral
b. Alternative – vancomycin or cephalosporine
c. Duration of treatment – 4 to 6 weeks
2. Methicillin resistant S. aureus (MRSA)
a. Vancomycin alone or in combination with
rifampin
 PREVENTION
• Staph infection will never be controlled because of the
carrier state in humans
• Home and hospital setting
• Proper hygienic care
• Disposal of contaminated materials
• Hospital setting
• Segregate persons with staph lesions
• Avoid indiscriminate use of antibiotics to prevent
establishment and spread of resistant strains
• Observe aseptic techniques
 PREVENTION
• In the newborn infant:
• Proper care of the umbilical stump
• Screen personnel in the nursery for staph carriers
STAPHYLOCOCCUS
EPIDERMIDIS
 EPIDEMIOLOGY

• Host specific for humans

• Most frequent sites – axilla, heads, arms,
nares, legs
• Nosocomial
• Resistant to many antibiotics like methicillin
and Pen G
 PATHOGENESIS

• Normal host – S. epidermidis has low

vilurence
• Distinct predilection for foreign bodies like
artificial heart valves, indwelling intravascular
catheters, CNS shunts and hip prostheses
 CLINICAL MANIFESTATIONS
• Causes infections of pacemakers, vascular grafts and
prosthetic joints and also peritonitis undergoing peritoneal
dialysis
• Single most common organism infecting intravenous
catheters
• Bacteremia
• UTI – elderly hospitalized men
• Natural valve endocarditis in intravenous drug abusers
• Produced toxins involved in Toxic Shock Syndrome
 TREATMENT

• Choice of appropriate therapy – based on the

local antibiogram
• Initial regimen – if no antibiogram
• Aminoglycoside (gentamicin or tobramycin)
with cephalotin + penicillinase resistant
penicillin
• Vancomycin alone or vancomycin +
aminoglycoside
 PREVENTION
• Difficult due to ubiquitous nature of organism
• Measures taken:
• Handwashing
• Reducing transmission rate from staff to patient or
patient to patient
• Proper surgical techniques help minimize
infections associated with installation of indwelling
medical device
STAPHYLOCOCCUS
SAPROPHYTICUS
 STAPHYLOCOCCUS
SAPROPHYTICUS
• Coagulase negative
• Resistance to novobiocin
• Failure to ferment glucose anaerobically
• Nonhemolytc
• Does not contain Protein A
• Common cause of UTI in sexually active young
women, second to E.coli – upper urinary tact is
involved
 STAPHYLOCOCCUS
SAPROPHYTICUS
• Selectively adheres to urothelial cells via specific
oligosaccharide receptors on cell membrane
• Certain strains are able to suppress growth of other
bacteria such as N. gonorrhea and S. aureus
attributed to an extracellular enzyme complex