GENUS STAPHYLOCOCCUS

The name staphyle (in Greek means bunch of grapes) is due to their appearance as
clusters as a result of division in many planes during replication.
Representants
It comprise more than 30 species, but 3 are medically important:
S.aureus responsible for most staphilococcal infections in humans
S.epidermidis causes opportunistic infections in immunosuppressed patients.
S.saprophyticus is opportunistic, cause urinary tract infections in women.
Differentiating characteristics of staphylococci
Characteristic
Color of colonies

S.aureus

S.epidermidis

yellow or white

white

S.saprophyticus
white or pale gray

Hemolysis

+/-

Coagulase production

Mannitol fermentation

Novobiocin sensitivity Sensitive

Sensitive

Cell wall teichoic acid

glycerol

ribitol

Resistant

S.aureus peptidoglycan is unique because of the pentaglycine bridges linking the

tetrapeptides.
S. aureus
Epidemiology
S.aureus colonizes the skin and mucous membranes of approximately 30% of normal
humans. (80-90% in healthcare personal). The anterior nares is the most commonly
populated site
-are implied in a lot of human infections
-human-to-human transmission is most common but nosocomial infection can also
occur (in hospitals to immunosupressed hosts and newborns)
-catalase positive
-prefer aerobic conditions but may behave as facultative anaerobes
-grow in presence of 7.5% sodium chloride (halotolerants)
Pathogenesis
Factors determining the occurrence of disease depend on:

-host: hormonal changes, devitalised tissues (wounds, burned areas, diabetus

mellitus), immunosupression due to iradiation, viral diseases, corticosteroids,
citostatic drugs, prolonged antibiotherapy.
-bacteria (S.aureus): number, determinants of pathogenicity
Determinants of pathogenicity
I.Surface (implied in the atachement of the staphylococcus to the host tissues)
-teichoic acids, peptidoglycan, capsula (present in young staphylococci), protein A
(covalently bound to the peptidoglycan layer of more than 90% of S.aureus isolates; it
binds to the Fc portion of Ig G, preventing specific antibodies from binding to the
bacteria and hindering Fc-mediated opsonization => complement is activated by the
protein A-bound immunoglobulins, which can contribute to a vigurous inflammatory
reaction.
II. Enzymes (determining both the invasive capacity and lesions)
*Coagulase = the most important marker of pathogenicity
-it helps the transformation of fibrinogen in fibrin
-2 tipes: free causes clotting of plasma
Linked forms a fibrine blanket over the group of staphilococci preventing
fagocytosis
*Fibrinolysin (staphylokinase)
*Hyaluronidase
*DNA-se
*Phospholipase: favour the instalation of staphylococcus in sebaceum regions where
they degrade lipids in fat acids, which can be used as carbon and energy sources. For
this reason they are also called FAME= fatty acids modifying enzymes)
*-lactamase (penicillinase)- they are easily transmitted from one strain to another
spreading the resistance to penicilins and other -lactamines.
III.Exotoxins
a.Pyrogenic exotoxins
-act as superantigens (the interaction with LT is not specific so that large numbers of
LT are implied, secreting a large amount of lymphokines).
1.Enterotoxins (A, B, C (1,2,3), D, E, F)
-heat stable proteins resistant to digestive enzymes
-responsible for the majority of food poisoning cases (in US and all over the world)

2.TSST-1 (toxic shock syndrome toxin-1)

-causes fever, multiple organ dysfunction and shock
-it is nearly identical with enterotoxin F
b. Citolytic exotoxins
1.Leukocidin kills PMN and macrophages
-immunogenic, because antibodies against this factor protect the host against
reinfection.
2.Hemolysins (,,, ) lyse erythrocytes
c.Exfoliatins
-cleave the stratum corneum, causing separation and loss of the most superficial
layers of the epidermis
-they are immunogenic => antibodies have been detected in recovering patients.
Clinical disease is primarily associated with S.aureus
I. Superficial infections occur most frequently and are characterized by intense
suppuration, local tissue necrosis and the formation of a pus filled local abscess.
1.Pyoderma (impetigo):
-etiology: S.aureus, S.epidermidis or Streptococcus
-highly communicable superficial skin infection: vesicles filled with clear to
yellowish fluid, most often on the extremities following infections of previous
mosquito bites or skin trauma.
2.Folliculitis, furuncles (boils) and sties result from infection of the hair follicles
3.Abscesses and carbuncles
II. Deep infections
1.Osteomyelitis
2.Pneumonia
3.Acute endocarditis
4.Arthritis, bacteremia, septicemia and deep organ (brain, kidney, lung) abscesses
III. Staphylococcal toxin diseases
1.Scalded skin syndrome
-manifestation of an exfoliatin producing strain of S.aureus
-involves infants and children younger than 5 years of age.
a)Bullous impetigo: few large localized blisters

b)Staphylococcal scarlet fever: erythematous rash but do not involves tongue

and palate
2.Staphylococcal food poisoning
-explosive vomiting and diarrhea and NO fever !!!
-occurs 1-5 hours after ingestion of contaminated food (e.g. meat, cream-filled
pastries, mayonnaise-containing salads)
-self-limiting, with proper hydration complete recovery occurs within 24-48 hours.
3.TSS
-febrile illness that may progress to organ failure and death.
Laboratory diagnosis
A. Direct diagnosis
1.Sample collection: pus, blood, nasal and pharyngeal exudates, urine, etc.
2.Microscopic examination: Gram smear : clusters of gram positive cocci on a
background formed by fibrin, necrotic descoamated cells.
3.Inoculation on media
-grow well under condition that are usually inhibitory to other bacteria (e.g. 7.5%
sodiumchloride or 40% bile)
-blood agar preferred for isolation from samples with little or no other microbial
contamination
-selective media with 7-10% sodium shloride, polymyxin, or bile.
-manitol salt agar (Chapman media) is selective and differential for staph.because
S.aureus ferments manitol => yellow colonies, whereas S.epidermidis does not =>
unmodified colonies.
Staphylococcus grow best at 37 C but become pigmented best at 20-25 C
4.Identification based on:
-morphology and staining: G (+) cocci arranged in clusters, non-motile, nonsporulated, the young stapylococci are capsulated
-cultural properties: in liquid media produce an uniform opalescence and on solid
media grow as big (2-3 mm), smooth, shiny colonies (S) with different pigmentation
depending on the species: white (S.epidermidis), white or yellow (S.aureus) and white
or gray (S.saprophyticus).
S.aureus produce hemolysis (incomplete = ), which can transform into complete
hemolysis () at 4 C.

-biochemical properties: catalase +, coagulase + (S.aureus), manitol fermentation and

oxidation + (S.aureus)
-pathogenic properties (production of toxic and non/toxic determinants of
pathogenicity)
a)in vitro

Coagulase test: a suspension of S.aureus is mixed with a drop of plasma, fibrin

clots form almost immediately.

Fibrinolysin test: in a media with fibrin they determine a clear area where the
colony grow
b)in vivo

Hemolysin is dermonecrotic: injected in rabbit (48-72 h) => lesion

Exfoliatin will cause exfoliation at the inoculation place in suckling mice.

Enterotoxin Dolman test: filtrate of a bacterial culture boiled for 20 min. is

injected intraperitoneal in kitten; after 10 to 60 minutes the kitten become agitated
has sialorhea, abdominal contractions, diarrhea. After 1 hour everything becomes
to normal.

5. Sensitivity to antibiotics and treatment

-must be performed because the resistance is very frequent and easily acquired from
one strain to another
-if treatment has to be done before isolation Oxacillin, Nafcillin and Meticillin are the
drugs of choice. For meticillin resistant strains vancomicin is preffered.
Cephalosporins, erythromycin or clindamycin may be used for patients allergic to
penicillin.
Control and prevention
Suppressing the carrier state population and implementing diligent aseptic practices
are the most effective measures of infection control. No vaccine is available.