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5 GRAM- Positive- Cocci- Staphylococci

Medical Technology (Our Lady of Fatima University)

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BACTERIOLOGY PROPER Staphylococcus spp.

I. Coagulase Positive – Staphylococcus
Group Bacteria Based:
aureus
II. CoNS – Coagulase Negative
1. Gram Positive Cocci
Staphylococci
2. Gram Negative Cocci
3. Gram Negative Bacilli
Positive Coagulase Negative Coagulase
4. Gram Positive Bacilli
Staphylococcus aureus Staphylococcus epidermidis
5. Miscellaneous Bacteria
- Pigment producer - - Normal flora of the skin
Kingdom Monera golden yellow Staph. saprophyticus
Family: Family: - Pathogenic to man. - Predominating normal flora
Micrococcaceae Sreptococcaceae of genitourinary tract (One
Staphylococcus hyicus of the most common cause
Genus Genus
- Pathogen to animal of UTI)
Micrococcus Streptococcus
Staphylococcus Enterococcus Staphylococcus albus
Staphylococcus
Stomatococcus Aerococcus - Pigment producer - white
intermedius
Planococcus Leuconostoc colonies.
- Pathogenic to animal
Lactococcus Staphylococcus citreus
Alloiococcus - Lemon yellow colonies,
Gemella pigment producer.
Pediococcus
Staphylococcus lugdunensis

GRAM POSITIVE COCCI

Point of Difference Micrococcus spp. Staphylococcus spp.

In pairs, tetrads, irregular

Morphology Gram Stain In cluster
clusters
Growth Chacteristics (5%
Obligate Aerobe Facultative anaerobe
Sheep’s Blood Agar)
Size Small to medium Medium to large
Transparency Opaque, convex Translucent, smooth, slightly raised
Hemolysis Non - hemolytic B – hemolytic and non hemolytic
Wide variety (white, tan, Yellow (golden/lemon), white, others
Pigment produced
yellow, pink) non – pigmented
Catalase Positive Positive
O/F Test Non – Glucose fermenter (-) Glucose fermenter (+)
(Oxidation/Fermentation) Glucose oxidizer (+) Glucose Oxidizer (+)
TESTS

Modified Oxidase Positive Negative

ANTIBIOTICS

Lysostaphin Susceptible Resistant

Furazolidine (100 ug disk) Susceptible Resistant
Bacitracin (0.04 ug disk) Susceptible Resistant

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Staphylococcus aureus Staphylococcus spp. Identification

 Pathogenic 1. Cultural Characteristics

 Produces substances ̵ Pinhead
̵ Media used:
Staphylococcus aureus Virulence Factor  BAP – S. aureus – beta
hemolytic
 Hemolysins (alpha, beta, gamma) – lysing RBC  NA
 Toxins  PEA Selective for Gram
1. Leukocidin / Panton – Valentine Factor  CAN positive bacteria.
– responsible to destroy WBC.
2. Exfoliatin – skin disease, enterotoxin F  PEA – Pheylethyl alcohol agar
3. Enterotoxin A to D – food poisoning  CAN – Columbia-colistin nalidixic acid agar

 Fibrinolysin/Staphylokinase – dissolving clot 2. Gram Stain

 Protein A ̵ Positive (VIOLET)
Anti - phagocytic
 Polysaccharide A ̵ Arranged in cluster
 Coagulase – abscess formation, clot –
formation. 3. Catalase Test
̵ Differentiate Staphylococcus
 Spreading Factor form Streptococcus.
̵ Hyaluronidase ̵ Used 3% Hydrogen Peroxide
̵ Lipase (H2O2)
̵ DNAse ̵ Formation of
 Other enzymes produced: Gelatinase, bubbles/effervesces.
Thermonuclease, Penicillinase, Catalase
4. Coagulase Test
Diseases Caused by S. aureus ̵ Differentiate S. aureus from
other non – coagulase positive
A. Local Purulent Infections Staphylococci.
̵ Boils ̵ Single bets pathogenicity for
̵ Wound infections Staph.
̵ Furuncles
̵ Carbuncles 5. Slide Method Tube Method
̵ Bullous impetigo - Detects - Free coagulase
̵ Sinusitis – sinus inflammation unbound - Confirmatory
̵ Otitis media – infection of the middle coagulase method
ear (clumping factor - Incubate 4 hours,
̵ Post – influenza pneumonia – small ball) 350C – 370C
- Screening - Reincubate in 18 –
B. Sepsis method 24 hours (to
C. Toxin – Caused Illness - Used plasma finalize the report)
̵ Food poisoning - Best source: - Result:
̵ Toxic Shock Syndrome Rabbit’s plasma Clot formation
̵ Scalded Skin Syndrome – common to - Result:
infant. Agglutination
Mannitol Fermentation
̵ Fermenting mannitol presence
in MSA.
̵ Used MSA as a selective and
differential.
̵ Positive: Yellow colonies of
S.aureus.

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CLOSED SYSTEM OPEN SYSTEM

6. DNAse Test  Added mineral oil  Oxygen free can
̵ Positive S. aureus - Devoid to be in utilize by the
̵ DNA medium – look-like NA contact in bacteria.
̵ Positive: Clear zone colonies. atmospheric  After incubation:
oxygen. Positive Result:
7. Gelatin Liquefaction Test  Positive Result: Yellow
̵ To determine the ability to produce Yellow acidic - Bacteria used
enzyme gelatinase. - Used glucose by glucose by
̵ Used 12% gelatin. fermentation. oxidation.
̵ Positive: S. aureus giving liquefied  Negative Result:
result. Green as is.
̵ Refrigerate for 4 hours:
o If still the same  POSITIVE
o If solidify  Negative

8. Novobiocin Susceptibility Tests

S. S.
S. aureus
epidermidis saprophyticus
Coagulase Positive Negative Negative
Novobiocin Susceptible Susceptible Resistant
Polymyxin Resistant Susceptible Susceptible

To differentiate the pathogenic from non-

pathogenic

9. Oxidation/Fermentation Test
̵ Differentiate Staphylococcus from
Micrococcus.
̵ Components:
 CHO: Glucose
 Peptone
 pH Indicator: Bromthymol Blue
̵ Staphylococcus: Positive Fermenter and
Oxidizer  Both yellow
̵ Micrococcus: Negative Fermenter (In
closed system) and Positive Oxidizer (in
open system)  Yellow and green

10. Modified Oxidase Test / Microdase Test

̵ Differentiate Micrococcus from
Staphylococcus.
̵ Cytochrome oxidase –
produced by Micrococcus
responsible for positive result:
Purple – Black Reaction.
̵ Active Chemical Component:
Tetramethyl – para –
phenylenediaminedihydrochlroride.

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