University of Baghdad

College of Medicine
2022-2023

Title: Pyogenic Cocci: Genus Staphylococci

Grade: 3rd
Module: Infectious Diseases Module
Speaker: Sarmad Zeiny
Date: 9 – Oct - 2022
Structure of bactria include
The enevlope(consist of capsule,cell wall and inner (cell or
palsma) membrane)

Pyogenic Cocci
The core(ribosomes,nucloid)
Appendages (flagella,pili(fimbria),cilia)
The diffrence between gram negative and positive cell
wall structre:
Gram positive/thick peptidoglycan layer interspersed
with techoic acid and project polysaccharides
Immediately beneth it is plasma membrane(here we don’t
call it inner membrane)both the techoic(lipotechoic acid)
and the special polysaccharides act as surface antigens
Gram positive cocci
also the whole peptidoglycan layer which is a
polysaccharide layer in addition to it forming the
exoskeleton it also act as an antigenic structure eleicting
production of various interlukins
Genus Staphylococci
Gram negative/by order Outer leaflet

1- outer membrane (normal plama membranw n term of Main classification according to shape is
phospohlipids)project polysaccharides (lipo ps) and 1-cocci(sphere)
interspersed with porin ‫االوتر ليفليت تحتوي‬

2-periplasmic space contaning thin layer of

2-bacili(rods) ‫نوعتوكسك من الليبد هو‬
A ‫الليبيد‬
Inner leaflet

pepitdoglycam and periplasmic gel 3-sprirlla(spirals) Phospolipid

inner leaflet‫عادي بال‬

3-inner membrance(plasma membrane)

Department of Microbiology, College of Medicine, University of Baghdad

Dr. Sarmad M.H Zeiny (M.B.Ch.B, M.Sc., F.I.C.M./Path)
‫اكو بالبوزتيف بس صغيرة كلش‬

outer leaflet or lps‫مكونات ال‬

‫هي‬
1-lipid A
2-core polysaccharide chain (with inner and outer core)
3-oligo- polysaccharide antigenic side chain
OBJECTIVES:
Upon completion of this lecture, the student will:

 Outline the medically important Staphylococci species.

 Describing the morphology & physiology for Staphylococci.
 Determine the virulence factors for Staphylococci.
 Analyze the diseases & pathogenicity for Staphylococci.
 Demonstrate the epidemiology/transmission for Staphylococci.
 Outline the laboratory diagnosis for Staphylococci.
 Clinically important Gram +ve cocci

S.Pyogenes S.aureus
S.agalactia
Streptococci Staphylococci S.epidermidis
S.Viridans
S.saprophyticus

S.Pneumococci
Staphylococcus
lugdunensis
S.enterococci
Staphylococci (Gram+ve cocci , catalase +ve)

Staphylococci
(Gram+ve cocci ,
catalase +ve)
 Clinical case:
 A box of ham sandwiches with mayonnaise prepared
by a person with a boil on his neck was left out of the
refrigerator for the on-call interns.Three doctors
became violently ill approximately 2 h after eating
the sandwiches. Dx.?

Dx.The most likely diagnosis is: food poisoning by

eating contaminated mayonnaise with toxin from
infected person with a boil which caused by
Staphylococcus aureus bacteria.
Genus Staphylococci divided in general into 2 types:

COAGULASE POSITIVE STAPHYLOCOCCUS

(PATHOGENIC)

Staphylococcus aureus

COAGULASE NEGATIVE STAPHYLOCOCCUS

(OPPORTUNISTIC) Are normal micriobiotaa

- Staphylococcus epidermidis 75%

- Staphylococcus saprophyticus
- Staphylococcus lugdunensis
Important properties of genus staphylococcus:
• The Shape: Staphylococci are spherical gram-positive cocci
• Arrangement: arranged in irregular grapelike clusters (Figure 1).

Grape like

• Biochemically:
- All staphylococci produce catalase enzyme.
-S.aureus produces coagulase enzyme  called coagulase positive staphylococcus. Non-flagellate
-S.epidermidis and S.saprophyticus are coagulase-negative staphylococci.
Continuation Important Properties….
• All Staphylococci tolerate high salt concentration (up to 9.5% Nacl).
Condiions for genus
staphylococcus to from
• Staphylococci are relatively resistant to drying, heat (they withstand 50°C for 30 pigments:
1.prolonged incubation
minutes), but are readily inhibited by certain chemicals (eg, 3% hexachlorophene). 2.room temperature
3.aerobic coditions

• S.aureus produces a pigment called staphyloxanthin, which imparts a golden color to 4.solid media

its colonies.
• S. aureus usually ferments mannitol and hemolyze RBC (β-hemolysis), whereas the
others do not.
• Most S aureus strains of clinical importance have polysaccharide capsules.
• S.saprophyticus resistant to novobiocin (type of antibiotic) while others are sensitive.
• All Staphylococci are non-motile and do not forms spores.
Staphylococcus aureus (Staphylococcus pyogenes):
Distinguishing features:
• Small, yellow colonies on blood agar.
• β – hemolytic on blood agar (complete destruction of RBC).
• Coagulase positive (all other Staphylococcus species are negative).
• Ferments mannitol on mannitol salt agar.
Reservoir:
- Human nasal mucosa (25% of population are carriers).
- Human Skin.

Transmission:
• Hands
• Sneezing
• Surgical wounds
• Contaminated food:
- Custard pastries
- Potato salad
- Canned meats
 ‫كل كرام بوزيتف بالسيل وول مالها اكو‬
‫باالضافة للتيكويك اسيد مجموعة بولي‬
‫ساكرايدز‬

Antigenic Structure:
‫ تصنعة‬exotoxin ‫الكرام بوزيتيف كلها تسوي‬
‫جوة جسمها وهي حية داخلك وتطلعة‬

1-Peptidoglycan: It elicits production of interleukin-1 and opsonic antibodies

‫فد شي يصعد درجة الحرارة‬

by monocytes, chemoattractant for leukocytes, and activate complement.

2-Teichoic acids: Antiteichoic acid antibodies.

3-Protein A: Is a cell wall component of S. aureus strains. Elicit formation of

anti-Protein A antibodies.

4- Clumping factor binds nonenzymatically to fibrinogen and platelets,

yielding aggregation of the bacteria. Differ from coagulase by being
antigenic while coagulase is not.
 Virulent factors and pathogenesis:
a) Proteins That Disable Our Immune Defenses (cell wall
@)Clumping factor is cell wall bound and is another example of an MSCRAMM that is responsible for adherence of the organisms to fibrinogen,fibrin and platelets . When mixed with plasma,

factors): S. aureus forms clumps. Clumping factor is distinct from coagulase. Because clumping factor induces a strong immunogenic response in the host,because after binding nonenzymetically to
fibrinogen and platlets it yields (resultys) in bactrial aggregation forming clumps

Enzymaticlly bound

1) Protein A: protect the organism from opsonization and phagocytosis. Bind to fc-
portion of IgG.
2) Coagulase: This enzyme can lead to fibrin formation. Coagulase binds to prothrombin; together they become enzymatically active and initiate
fibrin polymerization

3) Hemolysins: They destroy RBC, neutrophils, macrophages, and platelets.

4) Leukocidins :They destroy leukocytes. (Panton-valantine
leukocidine)
5) Penicillinase: This is a secreted form of beta-lactamase.
6) Novel penicillin binding protein (transpeptidase): Some strains of Staphylococcus
aureus have new penicillin binding proteins that are resistant to (penicillinase-resistant
‫الصفحة الجاية‬
‫مشروحة بشكل افضل‬

penicillins and cephalosporins).

7.Catalase
Staphylococci produce catalase, which converts hydrogen peroxide into water and oxygen. The catalase test differentiates
the staphylococci, which are positive, from the streptococci, which are negative.
b) Proteins (enzymes) to Tunnel Through Tissue:
1)Hyaluronidase ("Spreading Factor"): This protein breaks down
proteoglycans in connective tissue.
2)Staphylokinase: This protein lyses formed fibrin clots (like
streptokinase).
3)Lipase: This enzyme degrades fats and oils, facilitates S. aureus
colonization of sebaceous glands. Also called beta lactmase

4)Protease: destroys tissue proteins.

 (Include Tsst,exfolatin and enterotoxins)

Cornuem
‫طبقة مموجودة هنا الن الجلد مو‬
‫حيل ثخني هنا بس هي طبقة‬
‫ثانويك طالعة من الكورنيم اسمها‬ Granulusom(intense staining granules)
‫لوسيدم‬

‫فايروس يكودة ويطلعك على‬

‫سطح البكتريا‬
Comsisg of two Super antigens :

c) Exotoxin: 1. Exfoliative toxin A is encoded by eta located on a phage and is heat stable (resists boiling for 20 minutes).
2. Exfoliative toxin B is plasmid mediated and heat labile.

1) Exfoliatin: A diffusible exotoxin that causes the skin to slough off  ( SSS “scalded
skin syndrome”).
2) Enterotoxins (heat stable): Exotoxins which cause food poisoning.
3) Toxic Shock Syndrome toxin (TSST-1):
- Causes toxic shock syndrome.
- Found in 20% of S. aureus isolates.
- Superantigens which bind to the MHC class II.
- Causes a massive T-cell response and outpouring of cytokines.
Diseases:
 ‫متواجدة بكثرة‬

Diseases caused by direct organ invasion by Staphylococcus

aureus. Visualize the Staph-wielding wizard. (Note the
cluster of staphylococci at the head of his staff.)
 Another word for thicking or solidifying

Staphylococcus aureus Toxic shock syndrome caused by

gastroenteritis. Staphylococcus aureus releasing TSST-1.
Antibiotics VS Staphylococcus aureus
 MRSA=methicillin resistant staphylococcus aureus
Why S. aureus regarded as
multidrug resistant bacteria
(MDR)?
Staphylococcus aureus antibiotics resistance:
 90% of S.aureus produces β-lactamase (penicillinase) degrades penicillins.  Rx.
With β-lactamase–resistant penicillins, e.g., methicillin, nafcillin or cloxacillin. Plasmid
controlled.
 Methicillin-resistant S.aureus (MRSA) & Nafcillin-resistant S.aureus (NRSA).
- 50% of S.aureus strains isolated from hospital patients
- Penicillin-binding protein (PBP) changed, chromosome controlled.
- Rx. With vancomycin.
 Resistance to vancomycin, Intermediate (VISA) and full (VRSA):
- (VISA) : the mechanism of resistance is associated with increased cell wall synthesis
and alterations in the cell wall
- (VRSA): caused by the van genes acquired from enterococci. Plasmid controlled.
Staphylococcus aureus antibiotics resistance:
 Plasmid-mediated resistance to tetracyclines, erythromycins,
aminoglycosides, and other drugs is frequent in staphylococci from other
bacteria.

Note: MRSA, NRSA, VRSA & VISA can be treated with

either Linezolid, quinupristin/dalfopristin, daptomycin and
ceftaroline, a new cephalosporin with activity against MRSA.
 Quiz: T or F
1. S.aureus is a Gram –ve cocci arranged in grape like clusters.
2. All staphylococcus species are coagulase positive.
3. S.aureus can ferment mannitol.
4. MRSA is a staphylococcus species sensitive to penicillins.
5. S.aureus produces exotoxin and Protein AB.
 Staphylococcus epidermidis (Staphylococcus albus):
 Normal bacterial flora of the human skin.
 It is coagulase-negative.
 Infections are almost always hospital-acquired:
 Diseases: Infections of prosthetic devices in the body,
-Most frequent organism,
- Polysaccharide capsule (slime layer) that allows adherence. (regarded as
virulence factor)
 Highly antibiotic resistant. β-lactamase, The drug of choice is vancomycin.
 Staphylococcus saprophyticus
 Normal flora of the female genital tract and perineum. It has been isolated from
other sources too including meat and cheese products, vegetables, the
environment, and human and animal gastrointestinal tracts.
 This organism is a leading cause (second only to E.coli) of urinary tract
infections (UTI) in sexually active young women "Honeymoon cystitis".
 It is most commonly acquired by females in the community (NOT in the
hospital).
 This organism is coagulase-negative.
 Resistant to novobiocin.
 Staphylococcus lugdunensis
• Coagulase-negative staphylococcus.
• Colonies of S. lugdunensis are usually hemolytic, sticky, yellow or tan.
• S. lugdunensis may produce a bound coagulase (that is, the enzyme is bound
to the cells), a property it shares with S. aureus, but unlike S. aureus, it does
not produce a free coagulase. In the laboratory, it can give a positive slide-
coagulase test but a negative tube-coagulase test. P
• It is fairly easy to identify because, unlike the great majority of staphylococci,
it decarboxylates ornithine and is positive for pyrrolidonyl arylamidase
(PYR test).
• In the past, it was frequently misidentified as S. hominis, S. aureus, or other
species.
 Staphylococcus lugdunensis
• It occurs as a commensal on human skin, but has been recorded as a cause
of serious human infections such as osteomyelitis, arthritis, septicaemia,
wound infections, Acute postoperative endophthalmitis and aggressive
endocarditis.

• It is generally susceptible to antistaphylococcal antibiotics, but increasing

penicillin resistance has been reported.

• Lugdunin is an investigational antibiotic. Lugdunin is a non-ribosomally

synthesized cyclic peptide that inhibits growth of Staphylococcus aureus
strains.
Prevention:
 There is no vaccine against staphylococci.
 Clean, frequent hand washing, and aseptic management of lesions help to control
spread of S.aureus.
 Persistent colonization of the nose by S.aureus can be reduced by intranasal
mupirocin or by oral antibiotics, such as ciprofloxacin or trimethoprim-
sulfamethoxazole, but is difficult to eliminate completely.
 Shedders may have to be removed from high-risk areas, e.g., operating rooms and
newborn nurseries.
 Cefazolin (1st GC) is often used preoperatively to prevent staphylococcal surgical-
wound infections.
Laboratory & Self
study Direct
Specimens
Gram’s Slide
1
2

Biochemical Culture
4 3

Antibiotics
Serology
5
sensitivity
6
DR.SARMAD ZEINY,MBChB,MSc.
 Laboratory study
Steps of laboratory Diagnosis:
1) Specimens: according to type of infection, such as pus, blood, urine…etc.
2) Gram Stain: Gram’s positive cocci, Grape-like arrangement (clusters).
3) Culture: on both
- Blood agar: see large, round, smooth, raised & glistening white or golden color, β -hemolysis if
S.aureus.
- Mannitol Salt Agar: S.aurues: yellowish discoloration of media, other Staphylococci: no
discoloration (remains pink media)
4) Biochemical tests:
a) Coagulase test: is recognized as the most important test for testing the virulence of S.
aureus which is the only coagulase +ve staphylococci. So this test used to detect the ability of S.
aureus to clot blood plasma (fibrinogen fibrin). There are two main methods: Slide method and
tube method.
b) Catalase test: This is done to differentiate Staphylococci from Streptococci. A drop of 3% H2O2
solution placed on a slide, and a small amount of bacterial growth is in the solution placed on the
slide, the formation of bubbles indicates a positive test of oxygen release.
 Laboratory study

5) Serological test: Latex agglutination test to detect S.aureus. Serologic tests for diagnosis
of S aureus infections have little practical value.
6) Antibiotic Sensitivity test.
7) Molecular method like PCR. Polymerase chain reaction
8) For epidemiological purposes, S.aureus can be subdivided into subgroups based on the
susceptibility of the clinical isolate to lysis by a variety of bacteriophages. A person
carrying S.aureus of the same phage group as that which caused the outbreak may be the
source of the infections.
Summary:
 S.aureus is the most pathogenic species because of their
virulent factors.
 S.aureus colonizes nasal mucosa (25% of population are
carriers).
 S.aureus can cause simple and life threatening infections.
 Staphylococci easily diagnosed in the lab. By using Gram’s
staining, culture and biochemical tests.
 S.aureus and S.epidermidis are highly antibiotic resistance and
causing nasty hospital acquired infections.
 Staphylococci infection can be prevented by regular hygiene
precautions and by antibiotics, no vaccine.
THANK YOU