The Staphylococci

Morphology & Identification

(-)

(+)

Gram positive
Facultative anaerobes
Grape like-clusters
Catalase positive
Major components of
normal flora
skin
nose

Catalase test

Staphylococcus aureus

Antigenic Structure

Pathogenesis

Fc receptor
immunoglobulin

PHAGOCYTE
Protein A
BACTERIUM

Catalase
Coagulase
Hyaluronidase and Lipase
Hemolysin or sphingomyelinase C
Leukocidin
Exfoliative Toxin
Toxic Shock Syndrome Toxin
(superantigen)
Enterotoxins

Pathogenesis of staphylococcal infections

Stye:
Carbuncle:
Impetigo

Suppurative
Skin

Furuncle; Protein A, Leukocidin, Hemolysin

Stye; lipase
Impetigo; contagious
Epidermal necrolysis
Exfoliative Dermatitis (6,7,8); Exfoliative toxin
Mastitis
Abscess (deep tissue); granulation; coagulase, hyaluronidase (burn,
wound)

Systemic
Bactermia (from abscess, wound, burn) , Osteomyelitis (tibia) ,Pneumonia

 Food poisoning

not a human infection

food contaminated from humans
growth
enterotoxin
onset and recovery both occur within few hours
Vomiting/ nausea/ diarrhea/ abdominal /pain

Toxic shock syndrome

babies
scalded skin syndrome
* Exfoliatin

fever
scarlatiniform rash
desquamation
vomiting
diarrhea
myalgias

Infections associated with indwelling devices

Laboratory

1.
2.
3.

A. Direct examination; Gram Stain

B. Primary media; BAP
C. Differential Tests.
Mannitol Salts
Coagulase
DNase
D. Phage typing
E. Antibiotic Sensitivity (plasmid,
B lactamase) : penicillin
/methicillin/vancomycin

API STAPH Kit

Summary Figure (Identification Scheme)

Note: Strep. viridans
are alpha hemolytic and
negat ive for all the tests
below

GRAM POSITIVE COCCI

Catalase

Staphylococcus (Clusters)

Streptococcus (pairs & chains)

Coagulase

S. aureus
hemolytic
mannitol
yellow

Hemolysis

S. epidermidis
nonhem olytic (usua lly)
mannitol
(2)
white

BETA: Bacitracin

S .pyogenes (group A)

CAMP/Hippurate

S. agalactiae (group B)

ALPHA: Optochin/Bile Solubility

GAMMA: Bile Es culin

S. pneumoniae

+ 6.5% NaCl +

Group D*

Enterococcus

Bile Esc ulin

NaCl
Group D*
+ 6.5%
Non-Enterococcus

(*can also be beta or alpha hemolytic)

Staphylococcus epidermidis

major component skin flora

opportunistic infections
less common than S.aureus
nosocomial infections
heart valves
Identification
Non-hemolytic (sheep blood agar)
Does not ferment mannitol
Non-pigmented
Coagulase-negative

Staphylococcus saprophyticus
urinary tract infections
coagulase-negative
not differentiated from S. epidermidis

The Streptococcus

Streptococcus

Morphology & Identification

facultative anaerobe
Gram-positive
Chains or pairs
Catalase negative
(staphylococci are catalase positive)

Cell surface structure of S pyogenes

and extracellular substances

Lancefield groups
*one or more species per group
*surface antigens: M, T, R

groupable streptococci
A, B and D
most important
C, G, F
Rare

Non-groupable
S. pneumoniae
pneumonia
viridans streptococci
e.g. S. mutans
*dental
caries

Lipoteichoic Acid and F-protein

lipoteichoic acid
F-protein

fibronectin

epithelial cells

M protein
major target
natural immunity

strain variation
antigenicity

re-infection
occurs with different strain

M protein
IMMUNE

Complement
IgG

M protein

NON-IMMUNE

peptidoglycan

fibrinogen

Toxins & Enzymes

Hemolysis
alpha

beta

gamma

Classofication of Streptococci of Particular Medical Interest

Pathogenesis of S pyogenes infections.

S. pyogenes (Group A) -suppurative

affect all ages peak incidence at
5-15 years of age
non-invasive
pharyngitis
skin infection, impetigo

invasive bacteremia
toxic shock-like syndrome
"flesh eating" bacteria
pyrogenic toxin

Rheumatic fever -etiology

Scarlet fever
M protein
rash
cross-reacts heart myosin
erythrogenic toxin
autoimmunity
rheumatic fever
cell wall antigens
inflammatory disease
poorly digested in vivo
life threatening
persist indefinitely
chronic sequalae
fever
Post-infectious diagnosis (serology)
Heart
antibodies to streptolysin O
Joints
rheumatic NOT rheumatoid arthritis important if delayed clinical sequelae
occur
Acute glomerulonephritis
immune complex disease of kidney

superantigen
T cell mitogen
activates immune
system

Group B streptococcus identification

neonatal meningitis
septicemia
transmission
vaginal flora

hemolysis
hippurate hydrolysis
CAMP reaction
increases hemolysis of S. aureus

Group D streptococcus
Growth on bile esculin agar
black precipitate
6.5% saline
grow
enterococci
no growth
non-enterococci

Enterococci

distantly related to other streptococci

genus Enterococcus
gut flora
urinary tract infection
fecal contamination

opportunistic infections
particularly endocarditis

most common E. (S.) faecalis

resistant to many antibiotics
including vancomycin
terminal D-ala replaced by D-lactate

Viridans streptococci

diverse species
oral
dental caries
hemolytic and negative for other tests
non-groupable.
includes S. mutans
endocarditis

tooth extraction

Streptococcus pneumoniae

S. pneumoniae - diplococci

capsule:
pneumolysin:
Surface protein adhesinand
secretory IgA protease.
Teichoic acid and the
Peptidoglycan fragment,
phosphorylchorine .

leading cause pneumonia

particularly young and old
after damage to upper
respiratory tract
*e.g. following viral infection
bacteremia
meningitis
middle ear infections (otitis media)

Summary Figure (Identification Scheme)

Note: Strep. viridans
are alpha hemolytic and
negat ive for all the tests
below

GRAM POSITIVE COCCI

Catalase

Staphylococcus (Clusters)

Streptococcus (pairs & chains)

Coagulase

S. aureus
hemolytic
mannitol
yellow

Hemolysis

S. epidermidis
nonhem olytic (usua lly)
mannitol
(2)
white

BETA: Bacitracin

S .pyogenes (group A)

CAMP/Hippurate

S. agalactiae (group B)

ALPHA: Optochin/Bile Solubility

GAMMA: Bile Es culin

S. pneumoniae

+ 6.5% NaCl +

Group D*

Enterococcus

Bile Esc ulin

NaCl
Group D*
+ 6.5%
Non-Enterococcus

(*can also be beta or alpha hemolytic)

Bile solubility test

Streptex antiserum

optochin sensitive

Not optochin sensitive

Quellung reaction

using antisera
capsule "fixed"
visible microscopically

Latex agglutination - streptococci

Prevention and Treatment

Immunity ; 14 capsule types mixed
vaccine
Most strains susceptible to
penicillin , but resistance is
common

Neisseria

Gram negative

diplococci (pairs of
cocci)
oxidase positive

Culture: 5-10% CO2

Thayer Martin.
selective
chocolate agar
* heated blood

N. meningitidis

N. gonorrhoeae

Virulence Factors
Similar, but
Differences
in utilization

LPS

LPS

Capsule

IgA protease

Hemolysin
IgA protease
PILI
Opacity (OPA) proteins
Outer Membrane Proteins

PILI
Opacity (OPA) proteins
Outer Membrane Proteins

NO capsule
NO hemolysin

Neisseria gonorrhoeae
After 2-14 days
Found only in man
Gonorrhea: second most common venereal
disease
Gram stain of pure culture

Urethral exudate

Using the Gram stain in patient specimens, the organisms are most often
observed in polymorphonuclear leukocytes

Neisseria gonorrhoeae
Pili = key in anchorage of
organisms
to mucosal epithelium.
Nonpiliated gonococci are avirulent

OUTER MEMBRANE PROTEINS

Porin proteins (Por) = prevent phagolysosome fusion & allow
intracellular survival [ also called protein I]
Opacity proteins (Opa) = binding of organisms to epithelium
[also called protein II]
Reduction-modifiable proteins (Rmp) = protection against
bactericidal antibodies [ also called protein III]

Urethritis

Bartholins Duct

Purulent conjunctivitis/Ophthalmia neonatorum Infection in

newborns during vaginal delivery

Disseminated gonococcal infection

(DGI).
Fever, polyarthritis
(or monoarticular septic arthritis),
and/or dermatitis
(pustules on a hemorrhagic base).

Smear
polymorphonuclear cell
Gram negative cocci
many in cells
Culture

Antibiotic therapy
lactamase-resistant cephalosporin
e.g. ceftriaxone
resistant strains
common
produce lactamases
destroy penicillin

N. meningitidis
(the "meningococcus")

Neisseria
meningitidis
resides in man only
usually sporadic cases
mostly young
children
outbreaks
adults
crowded conditions
* e.g. army
barracks

Upper respiratory
tract infection
adhesion pili

Meningococcal
meninigitis
1-4 days
Second most common
meningitis
pneumococcus,
most common
Fatal if untreated
Responds well to
antibiotic therapy
penicillin

Bloodstream

Brain

Prevention
Diagnosis
spinal fluid
Gram negative
diplococci
within
polymorphonuclear cells
meningococcal
antigens

Culture
Thayer Martin agar

capsule
inhibit phagocytosis

anti-capsular antibodies
stop infection

antigenic variation
serogroups

vaccine
multiple serogroups