Gram positive cocci

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7 -11- 2010 2
Staphylococcus

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Morphology and Identification

 Gram positive, catalase positive cocci.

 Spherical cells, 1 micrometer in diameter.
 Arranged in grape like clusters.
 Non-motile, do not form spores.
 They are the most common cause of pyogenic infection.
 They are frequent isolates from the oral cavity.
 Higher proportions of Staphylococcus aureus are found
in the saliva of healthy subjects older than 70 years.

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Species of medical importance

 Staphylococcus aureus
 Staph. epidermidis
 Staph. saprophyticus

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Classification

 Based on tube Coagulase test they are

classified into:
1. Coagulase positive staphylococci -
Staphylococcus aureus
2. Coagulase negative staphylococci -
Staph.epidermidis & Staph.saprophyticus.

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Staphylococcus aureus

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Morphology and Identification

 Grow readily on most bacteriologic media

under aerobic or microaerophilic conditions.
 Grow most rapidly at 37 °C but form pigment
best at room temp. (20-25 °C(
 On solid media colonies are round, smooth,
raised, and glistening.
 S. aureus usually forms gray to deep golden
yellow colonies.

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staphylococcus

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Epidemiology
 Habitat (reservoir)
• Normal flora of human anterior nares,
nasopharynx, perineal area, and skin
• can colonize various epithelial or mucosal
surfaces.

 Mode of transmission
• Spread of patient’s endogenous strain to
normally sterile site by traumatic introduction
• Maybe transmitted person to person by fomites,
air, or unwashed hands of health care workers.
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Antigenic structure
Virulence factor:

 Capsule- polysaccarides antiphagocytic

 Peptidoglycan- activates complement and

induce cytokines.

 Tiechoic acid-for adhesion to host cell.

 Protein A- present in cell wall, antiphagocytic.

Enzymes and Toxins
 Catalase.
 Coagulase and clumping factor:
 Coagulase, an enzyme that clots plasma.
• it is secreted free into culture medium when
staph are grow.
• it is require plasma factor for it is clotting
action.
• coagulase converts fibrinogen to fibrin.

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Enzymes and Toxins

 Clumping factor is a surface S. aureus

compound that is responsible for adherence
of the organisms to fibrinogen and fibrin.

 Other enzymes: phosphatase and

deoxyribonucleases.

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 Exotoxins.
 Leukocidin : an important virulence factor in
community associated MRSA infections.
 Heamolysins- Alpha, Beta, Gama, Delta
lysins.
• Alpha toxin has cytotoxic,leukocidal & has
dermonecrotic action.
• It causes injury to circulatory system and
muscle tissue.
 Exfoliative toxins.
• Type A&B, they cause generalized
desquamation of skin known as
Staphylococcal Scaled Skin Syndrome
(SSSS).
• Toxin A is coded by chromosome,

• Toxin B is coded by plasmid.

 Toxic shock syndrome toxin (TSST-1).

Strain colonize in vagina.

Scaled Skin Syndrome
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 Enterotoxins.
• 8 type (A, B, C1, C2, C3, D, E&H).
• Type A is responsible most cases of food
poisoning.
• This toxin is resistant to gut enzyme and boiling
for 30minutes.
• food sources are contaminated with toxin but not
the bacteria.
• Mostly milk and milk products.
• the sources of infection is usually a lesion in
hand of food handler which contain staph..
Pathogenesis and spectrum of disease
 Rarely causes oral infections; may cause angular cheilitis
(together with the yeast Candida) at the angles of the
mouth

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Pathogenesis and spectrum of disease
 Staphylococci causes localised pyogenic lesions.

 Cutanoeus infections – pustules, boils, carbuncles,

furuncles, abscesses, impetigo, wound & burn infections

 Deep infections – Osteomyelitis, tonsillitis, pharyngitis,

sinusitis, pneumonitis, endocarditis, meningitis &
septicaemia.
 Can cause urinary tract infection in diabetic patients

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Carbuncles Furuncles

Impetigo
Boils
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Pathogenesis and spectrum of disease
 Food Poisoning – Occurs 2 to 6 hrs after ingestion of
contaminated food which contains preformed Enterotoxin

 Exfoliative skin lesions - bullous impetigo, pemphigus

neonatorum, staphylococcal scalded skin syndrome

 Toxic shock syndrome - fever, hypotension, vomiting,

diarrhoea & erythomatosus rash. It was associated with
the use of contaminated tampons by menstruating
women but occurs in other conditions also.

 Nosocomial infections – Methcillin resistant

Staph.aureus causes most of the nosocomial infections.22
Laboratory Diagnosis

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Specimens

 Surface swab pus.

 Blood.
 Tracheal aspirate.
 Spinal fluid for culture.
 Stool .
 Urine.

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Direct detection methods

 Gram stain:
 Spherical, gram-positive cells.

S. aureus

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Cultivation

 Media:

 5% sheep blood and chocolate agars but not

MacConkey agar.

 Broth-blood culture systems and common nutrient

broths, such as thioglycollate and brain-heart infusion.

 MSA (mannitol salt agar): to isolate staphylococci from

clinical material selectively, and screen for nasal carrier
of s.aureus and pt with cystic fibrosis.

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Incubation Conditions and Duration

 5% sheep blood and chocolate agars in CO2

or ambient air usually occurs within 2r hrs.

 MSA and other selective media may require

incubation for at least 48-72 hrs.

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Biochemical reactions

1. Catalase test-
• S. aureus colonies produce air bubbles when
mixed with 3% hydrogen peroxide indicating
positive test.
2. Coagulase test- +ve only s.aureus.
• slide coagulase.
• tube coagulase.
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Extracellular (free) Bound coagulase
coagulase
Clumping factor
10%-15% of S. aureus
are -ve

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• S. aureus
 Coagulase test +ve

 B-haemolysis +ve

 Pigment on media- golden to yellow pigment.

 DNAse test +ve

 Mannitol test +ve

 Phosphatase production +ve

 Bacteriophage typing

 About 23 bacteriophages are used.

 The strain of Staphylococci to be identified is grown on nutrient
agar.
 After drying, the phages are applied in fixed dose.
 After overnight incubation the phages capable of lysing are
designated to that strain.
 Example; Phage type 52/80/94 is lysed by phages 52, 80 & 94
only. Five types of phage groups can be identified.
 Group I 29, 52, 52A, 79, 80
 Group II 3A, 3C, 55, 71
 Group III 6, 42E, 47, 53, 54, 75, 77,83A, 84, 85
 Group IV 42D
 Group V 94, 96

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Phage typing

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Antibiotic sensitivity testing:
 Staph aureus are usually sensitive to penicillin but now
penicillin resistant strain are emerging.

 penicillin resistant: some strain produce B lactamase

enzyme which can inactivate penicillin by splitting B
lactam ring .this Enzyme are code by plasmids.

 MRSA
• Methicillin resistant is coded by Mec A gene which is
transmitted chromosomally.
Treatment
 Penicillin sensitive strains:
• Penicillin, erythromycin , gentamycin,
ciproflixacin.

 Penicillin resistant strains :

• cloxacillin,methicillin, oxacillin.

 MRSA
• vancomycin,teicoplanin,linzeolid.
Coagulase- negative staphylococci (CNS)

 The CNS are normal human flora and sometimes cause

infection,

 often associated with implanted appliances and devices,

especially in very young, old, and immunocompromised
patients.

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Coagulase- negative staphylococci (CNS)
 S. epidermidis
• Normal flora of skin and nose,

• coagulase negative

• cause opportunistic infection in pt with artificial valve,

catheter.

• It become pathogenic in immuncompromised individuals.

• 75% of infections caused by coagulase- negative

staphylococci are due to S. epidermidis.
Coagulase- negative staphylococci (CNS)

S. saprophyticus:

 coagulase negative

 an opportunistic pathogen causing UTI

(common in young women)

 can cause endocarditis & septicemia .

Micrococci

 G+ve cocci
 Catalase-positive
 Coagulase-negative and usually grow as white colonies
on blood agar
 Stomatococcus mucilagenosus, formerly classified in the
genus Micrococcus, is found in abundance on the lingual
surface.
 This species has the ability to produce an extracellular
slime, which correlates with its predilection for the lingual
surface.
 Its role in disease, if any, is unknown.

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Thank you

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