Staphylococcus spp.

Back then, Staphylococcus spp and Micrococcus spp belonged in the (blank) family, but because of molecular and chemical analysis, they have been separated

micrococcaceae

What is the phylum od Staphylococcus spp?

firmicutes

What is the phylum of Micrococcus spp.?

actinobacteria

What is the family of Staphylococcus spp

Staphylococcaceae

What are the two families of Micrococcus spp?

1. micrococcaceae

2. dermacoccaceae

Where can you usually see the normal flora of Staphylococcus spp?

1. skin

2. mucous membranes

Where can you usually see the normal flora of Micrococcus spp?

skin

Since Micrococcus is a normal flora of the skin, what are the 3 usually “skin colonizers”?

1. micrococcus spp.

2. kocuria spp.

3. kyrococcus spp.

What is the gram stain of Staphylococcus spp?

gram +

What is the morphology and arrangement of Staphylococcus spp?

cocci in singly, pairs, and are usually in grape-like clusters

What is the gram stain of Micrococcus spp.?

gram +

What is the morphology and arrangement of Micrococcus spp?

cocci in pairs, tetrads, and irregular clusters

Aerotolerance of Staphylococcus spp or Micrococcus spp?

• Facultative anaerobe (majority) may or may not require the presence of O2

• Obligate aerobe

Staphylococcus spp.

Aerotolerance of Staphylococcus spp or Micrococcus spp?

• Obligate aerobe since theey need the presence of O2 to perfrom their metabollic processes

Micrococcus spp.

Since one of the aerotolerance of Staphylococcus is obligate aerobe, there are exceptions where they do not want O2 for their survival, what are these?

1. S. aureus subsp. anaerobius

2. S. saccharolyticus

Since one of the aerotolerance of Micrococcus spp. is obligate aerobe, there are exceptions where they may or may not require the presence of oxygen, what are these?

1. M. kristinae

2. M. varians

Is Staphylococcus catalase + or -?

+

Does Staphylococcus spp. have bubble formation?

yes

What is usually the color of the colonies in Staphylococcus spp.?

yellow pigment

What is usually the colony size for Staphylococci

medium sized/pinhead

What is usually the density for Staphylococci

opaque

Positive or Negative:

• Staphylococci in modified oxidase/microdase test

negative

Positive or Negative:

• Micrococci in modified oxidase/microdase test

positive

Positive or Negative:

• Staphylococci in Anaerobic acid production from glucose test

positive

Positive or Negative:

• Micrococci in Anaerobic acid production from glucose test

negative

Positive or Negative:

• Staphylococci in Anaerobic acid production from glycerol in the presence of erythromycin test

positive

Positive or Negative:

• Micrococci in Anaerobic acid production from glycerol in the presence of erythromycin test

negative

Positive or Negative:

• Staphylococci in Furoxone-Tween 80-oil red O agar test which is a selective medium because of the inhibitor furoxone

negative, inhibited, no growth

Positive or Negative:

• Micrococci in Furoxone-Tween 80-oil red O agar test which is a selective medium because of the inhibitor furoxone

positive, not inhibited, has growth

Resistant or Susceptible/Sensitive:

• Staphylococci in resistance to bacitracin test (0.04U)

resitant

Resistant or Susceptible/Sensitive:

• Micrococci in resistance to bacitracin test (0.04U)

susceptible/sensitive

Resistant or Susceptible/Sensitive:

• Staphylococci in resistance to lysosome test (50 mg disk)

resistant

Resistant or Susceptible/Sensitive:

• Micrococci in resistance to lysosome test (50 mg disk)

susceptible/sensitive

Resistant or Susceptible/Sensitive:

• Staphylococci in resistance to lysostaphin test (200 ug/mL)

susceptible/sensitive

Resistant or Susceptible/Sensitive:

• Micrococci in resistance to lysostaphin test (200 ug/mL)

resistant

In greek, what does staphle mean?

bunch of grapes

• This spp is non-motile

• Non-spore forming

• No flagella

• Is considered as a fastidious strain that usually requires CO2, hemin, or menadione

Staphylococcus spp.

Staphylococcus have small colony variants, they grow slow about (blank) the size of the other strains when grown for 18 to 24 hours

1/10

In the macroscopic examination of Staphylococcus spp., what size it is usually?

medium sized about 4 to 8 mm

In the macroscopic examination of Staphylococcus spp., what does it appear like? what is the color?

creamy, colored-white, or light gold

In the macroscopic examination of Staphylococcus spp., what is the texture like?

buttery

In the microscopic examination of Staphylococcus spp., what is it arranged as commonly?

grape-like clusters

These two spp of Staphylococcus spp that are posseses polysaccharide capsule that may also appear as a slime layer which serves as a virulence factor that can also produce biofilm to adhere to inorganic surfaces

1. S. aureus

2. S. epidermis

What are the Staphylococcus spp. that are catalase (-) and are obligate aerobes

1. S. aureus subsp. anaerobius

2. S. saccharolyticus

What are Staphylococcus that are microdase (+)

1. S. scuri

2. Macrococcus caseolyticus

3. S. lentus

4. S. vitulus

What are the 8 virulence factors of Staphylococcus aureus

1. enterotoxins

2. toxic shock syndrome toxin-1

3. exfoliative toxin

4. cytolytic toxin/cytotoxin

5. enzymes

6. protein A

7. peptidoglycan

8. teichoic acid

• This is a virulence factor of Staphylococcus aureus

• Group A to E and G to J

• Responsible for Staphylococcal food poisoning, Staphylococcal pseudomembranous enterocolitis, diarrhea, vomitting,

• Considered as super antigens because they can easilt actvate T cells where we get excessive immune response from

• Is heat stable for 100 degrees for 3 minutes that is produced by 30 to 50%

enterotoxin

• This is a virulence factor of Staphylococcus aureus

• Also known as pyrogenic toxin and TSST-1

• TSS

• previously known as enterotoxin F

• Ias associated with fever, desquamation, and hypotension potentially leading to shock and death

• Has two types

toxic shock syndrome toxin-1

What are the two types of TSST-1?

1. menustration-associated

2. non-menustrating associated

In non-menstruating associated TSST-1, its caused by?

surgical infections

• This is a virulence factor of Staphylococcus aureus

• Also called as epidermolytic toxin

• Caused by epidermolytic toxin A and B

• Causes Staphylococcal SSS and bullous impetigo

exfoliative toxin

In exfoliative toxin, what does Staphylococcal SSS mean?

scalded skin syndrome

In exfoliative toxin, this is a disease that causes epidermal layer detachment producing a burning-like effect and is most common in <5 years old

bullous impetigo

This is a disease caused by exfoliative toxin, what do we call when the bullous impetigo is localized?

pemphigus neonatorum

This is a disease caused by exfoliative toxin, what do we call when the bullous impetigo is generalized? It covers several parts of the body and may be systemic

Ritter’s disease

• This is a virulence factor of S. aureus

• It is the extracellular proteins that affects red blood cells and leukocytes

cytolytic toxin or cytoxin

• This is a virulence factor of S. aureus called cytolytic toxin

• This is the portion of lysins and leukocidins

• What are the 4 hemolysins under S. aureus?

1. a

2. B

3. delta/o

4. y/non

In the cytolytic toxin of S. aureus under lysins and leukocidins, what hemolysin is this?:

• It disrupts the smooth muscles in the blood vessels and is extremely toxic to erythrocytes, leukocytes, hepatocytes, and platelets

a-hemolysin

In the cytolytic toxin of S. aureus under lysins and leukocidins, what hemolysin is this?:

• It is also known as sphingomyelinase or hot and cold lysin

• Has enhanced hemolytic activity when incubated at 37 degrees and subsequently at 4 degrees

• It also acts in CAMP test

B-hemolysin

In the cytolytic toxin of S. aureus under lysins and leukocidins, what hemolysin is this?:

• It is less toxic than a-hemolysin and B-hemolysin

delta/o-hemolysin

In the cytolytic toxin of S. aureus under lysins and leukocidins, what hemolysin is this?:

• This functions with the PVL to kill PMNs

y-hemolysin

What does PVL stand for?

Panton-Valentine Leukocidin

What are the 5 enzymes under the virulence factors of S. aureus?

1. coagulase

2. protease

3. hyaluronidase

4. lipase

5. B-lactamase

• This is an enzyme under the virulence factor of S. aureus

• Serves as the primary marker for virulence

• Promotes clot formation

coagulase

• This is an enzyme under the virulence factor of S. aureus

• It degrades proteins

• Aids in tissue destruction

protease

• This is an enzyme under the virulence factor of S. aureus

• Hydrolyzes hyaluronic acid present in the intracellular ground that makes up connective tissues which then promotes the spread of bacteria

hyaluronidase

• This is an enzyme under the virulence factor of S. aureus

• This is coagulase + and -

• It acts on lipids present on the surface of the skin

lipase This is an enzyme under the virulence factor of S. aureus

• This is an enzyme under the virulence factor of S. aureus

• This cleaves the ring structure of penicillin and other antibiotics therefore making them resistant

B-lactamase

• This is another virulence factor of S. aureus

• It is found in the cell wall

• Binds to the Fc portion of the cell wall to inactivate IgG

protein A

• This is another virulence factor of S. aureus

• This activates the complement IL-1 which is chemotactic to PMNs which leads to tissue damages

peptidoglycan

• This is another virulence factor of S. aureus

• This mediates binding to the fibronectin

teichoic acid

• This is another virulence factor of S. aureus

• Present only in S. epidermidis

polysaccharide capsule/slime layers

• This is another virulence factor of S. aureus

• Enhances adherence to host surfaces

• Enhances antibiotic resistance

biofilm

For the specimen collection of S. aureus, what is the most ideal sample?

aspirates

What can you see with the direct microscopic examination of S. aureus?

gram + cocci with PMNs

Where can you culture S. aureus?

1. BAP

2. CAP

3. MSA

4. CNA

5. PEA

6. CHROMagar Staph aureus

How long should S. aureus be incubated? and at what temp?

18-24 hours at 35 to 37 degrees

When you grow S. aureus on BAP, what do you see?

B or non-hemolytic pattern

MSA is a selective differential media because of what?

7.5 to 10% NaCl

What is the pH indicator for MSA?

phenol red

MSA inhibits what?

halophiles

What are the 4 spp. that can grow in MSA?

1. S. aureus

2. S. epidermidis

3. S. saphrophyticus

4. Micrococcus spp.

When you grow S. aureus in MSA what do you see?

yellow

When the growth in MSA is yellow, it means that it is a what?

mannitol fermenter

When the growth in MSA is red, it means that it is a what?

non-mannitol fermenter

When there is no growth in MSA, it means that it is a what?

non-halophile

When you grow S. epidermidis on MSA, what color do you see?

red

When you grow S. saprophyticus in MSA, what do you see?

yellow

When you grow micrococcus in MSA, what do you see?

no growth

• This media for S. aureus is only used when the sample is heavily contaminated

• This is a selective and enriched media

CNA

When using CHROMagar Staph aureus, what is color for S. aureus

mauve

When using CHROMagar Staph aureus, what is color for S. saprophyticus

turquoise blue

When using HardyCHROM, what is the color for S. aureus?

pink

When using HardyCHROM, what is the color for S. epidermidis?

turquoise blue

What is the reagent for catalase test?

30% hydrogen peroxide

Catalase test is sed to differentiate what?

Staphylococcus and Micrococcus from Streptococcus and Enterococcus

When the catalase test is +, meaning that there is bubble formation, what are the examples species?

Staphylococcus and Micrococcus

What does the microdase test differentiate?

micrococcus from S. aureus

What is the enzyme being detected in microdase test?

oxidase and cytochrome c

What is the color of the + result of microdase test?

blue or purple-blue color within 2 minutes