This week

Chapter 7 Coccus
Chapter 3 Disinfection & Sterilization
Exp. Effects of physical, chemical and
biological factors on the growth of bacteria
Chapter 7 Cocci
 Biological characteristics;
Outline Pathogenicity;

Pathogenic cocci (pyogenic cocci)

 G+ :
 Staphylococcus
 S.aureus
 Streptococcus
 S.pyogenes, S.pneumoniae
 Enterococcus (low virulent, normal flora)
 G- :
 Neisseria
 N.meningitidis, N.gonorrhoeae
Section 1 Staphylococcus
Staphylococcus
widely exist in nature
 Primary reservoir: animal and human body;
human --- the most common carriers;
skin, nasopharynx, throat
---main bacteria in the nosocomial infection.
>45 species
---S. aureus: the most virulent species.
---S. epidermidis: opportunistic pathogen.
---S. saprophyticus: rarely cause human disease.
community-acquired UTI.
 Biological characteristics

Shape and structure

 G+, round, arranged in
irregular, grape-like
clusters
 flagella(-), spores(-),
pili(-)
 Some species
 slime layer only in their
early age
 Biological characteristics
• Cultivation
low nutrient requirement
grow in basic media;
facultative anaerobes;
temperature: 37℃(28℃-38℃);
pH: 7.4(4.5~9.8);
10%~15% NaCl
 Biological characteristics
S.aureus
Growth properties
liposoluble pigments:
S.aureus: golden yellow color
others: white or citrine color
 Biological characteristics
S.aureus
Growth properties
complete hemolysis (-hemolysis)
(S.aureus)
 Biological characteristics
Staphylococci
Biochemical reactions Streptococci

Catalase test “+”

Staphylococci: “+”
Streptococci: “-”
 Biological characteristics
Biochemical reactions
Mannitol Fermentation “+”
S. aureus: “+”;
S.epidermidis & S.saprophyticus: “-”;
 Biological characteristics
• Antigenic composition

 Staphylococcal protein A (SPA)

cell wall protein
covalently bound to peptidoglycan
S. aureus
 Characteristics:

nonspecific combination with

IgG Fc except IgG3
Biological characteristics
SPA: virulent factor:
 a. anti-phagocytosis;
 b. damage platelet;
 c. induce hypersensitivity and inflammation;
 SPA(staphylococcal protein A)

Phagocyte

Fc receptor

IgG SPA

Fab Fc
Bacteria
 SPA(staphylococcal protein A)

Phagocyte

Fc receptor
×
IgG SPA

Fab Fc Ag
Biological characteristics
SPA: virulent factor:
 a. anti-phagocytosis;
 b. damage platelet;
 c. induce hypersensitivity and inflammation;
 Biological characteristics
SPA function in diagnosis:
SPA-coated S.aureus is used as reagent (Ab carriers) of
coagglutination test to identify unknown species and types of
bacteria.
 coagglutination test
reagent
SPA-coated S.aureus
(Ab carrier)

known Ab

unknown Ag
 Classification
Based on the pigments and chemical reactions: >45 species
Properties S. aureus S. epidermidis S. sarprophyticus
Pigment Golden yellow White White/Citrine
Coagulase + － －
Mannitol + － －
Thermostable nuclease + － －
β-hemolysis + － －
SPA + － －
virulent strong Weak -
Classification
Based on coagulase
 Coagulase” +”: e.g., S. aureus
 Coagulase” -”: e.g., S. epidermidis
S. saprophyticus
Resistance to environment
Resistance--higher than other non-spore forming bacteria
 Dryness
 Heat (survive at 80℃ 30 min);
 high concentration of salt (10-15% NaCl)
Sensitivity:
 basic dyes (e.g., crystal violet)
 several antibiotics
(e.g., penicillin, vancomycin)

MRSA (methicillin resistant S.aureus): >90%

 Pathogenicity (S.aureus)

• Virulent factors
 Invasiveness  Toxin---exotoxins
• Hemolysins
• Surface structures
• Leukocidin
SPA, slime layer
• Staphylococcal enterotoxin
Invasive enzymes
• TSST-1
Coagulase
• Exfoliative toxin
Enzyme

Coagulase:
 plasma clotting protein
soluble fibrinogen in citrated plasma
↓convert
insoluble fibrin
 2 types
Enzyme
Coagulase:
 2 types
Free coagulase
-- cause plasma clotting
-- detected by tube coagulase test
Bound coagulase
-- cause bacterial clump
-- detected by slide coagulase test
 Roles:
 inhibit the phagocytosis;
 inhibit function of bactericidal substances in human body by
coating bacteria with fibrin or formation of bacterial clump;
 make abscess localized
 Free coagulase

+ - control

Plasma clot

tube coagulase test

Bound coagulase

slide coagulase test

Pathogenicity (S.aureus)

• Virulence factors
 Invasiveness
• Surface structure
SPA, capsules, slime layer
Invasive Enzyme
Coagulase
 Toxins---exotoxins
• Hemolysins (Staphylolysins)
• Leukocidin
• Staphylococcal enterotoxin
• TSST-1
• Exfoliative toxin/
Epidermolytic toxin
 Hemolysins/Staphylolysins
4 kinds: -Lysin (main pathogenic substance)
-Lysin
-Lysin
-Lysin

Roles:
• damage host cell membrane  permeability;
• have cytotoxic effects on phagocytes and tissue cells;
e.g., erythrocytes, leukocytes, platelets, hepatocytes
Pathogenicity (S.aureus)

• Virulence factors
 Invasiveness
• Surface structure
SPA, capsules, slime layer
Invasive Enzyme
Coagulase
 Toxin---exotoxin
• Staphylolysin(Hemolysins)
• Leukocidin
lyse neutrophils and macrophages
Staphylococcal enterotoxin
TSST-1
Exfoliative toxin/
Epidermolytic toxin
Staphylococcal enterotoxin

~1/2 strains of S.aureus

Heat stable protein (100℃, 30 min)
Resistant to proteolytic enzymes of GI tract
Types: A, B, C, D, E, G
Role: stimulate vomiting-control center of CNS and cause
a food poisoning and colitis characterized by projectile
vomiting and diarrhea.
Toxic shock syndrome toxin-1(TSST-1)

Role:
 Cause TSS (Toxic shock syndrome)
 Induce abrupt high fever, shock, hypotension, multi-organs
failure;
 enhance the host susceptibility to endotoxin
Exfoliative toxin (Epidermolytic toxin)

Role: cause blister of skin, peeling of the skin

staphylococcal scalded skin syndrome (SSSS)
Pathogenicity
Diseases 2 types
Invasive infection (pyogenic/purulent infection)
skin infection (cutaneous and subcutaneous infecton):
 form localized abscess (Lesion is limited in local area)

 e.g., folliculitis; boil; carbuncle; impetigo, etc.

organ infection: pneumonia; empyema, meningitis, etc.
Systemic infection: bacteremia, septicemia; pyemia
folliculitis

Boil/furuncle
 Impetigo- staph skin
infection, multiple pustules
Carbuncle (small skin abscesses)
Pathogenicity
Diseases 2 types
Invasive infection (pyogenic/purulent infection)
skin infection (cutaneous and subcutaneous infecton):
 form localized abscess (Lesion is limited in local area)

 e.g., folliculitis; boil; carbuncle; impetigo, etc.

organ infection: pneumonia; meningitis, etc.
Systemic infection: bacteremia, septicemia; pyemia
 Pathogenicity
Toxin-associated diseases
 Food poisoning (enterotoxin)
 Short incubation period: 30min~4 hrs.
 Symptom: projectile vomiting, nausea, abdominal cramps; diarrhea; no fever.
 recover in 1-2 days without treatment.
 TSS (TSST-1)
 a multisystem disease
 sudden high fever, shock, hypotension, kidney failure, vomiting, diarrhea, and red
skin rash, etc.
 SSSS (Exfoliative toxin) (staphylococcal scalded skin syndrome)
 Staphylococcal scalded skin syndrome (SSSS)

-- most often occurs in newborns

and young children (<4 age).

-- low mortality rate.

skin blister →denudation (like scald)

Coagulase- staph.
Diseases:
 S.epidermidis
 urinary tract infection (UTI)
 bacterial endocarditis

 septicemia

 wound infection

 S.saprophyticus
 UTI
Staphylococcus aureus has been linked to all
of the following conditions EXCEPT ( )
 A. impetigo.
 B. the scalded skin syndrome.
 C. toxic shock syndrome.
 D. boils.
 E. Rocky Mountain Spotted Fever.
pathogenic cocci
3 Genera
 G+ cocci: Staphylococcus
S. aureus
Streptococcus
S. pyogenes
S. pneumoniae
 G- cocci: Neisseria
N. meningitidis
N. gonorrhoeae
Section 2 Streptococcus
-- widely exist:
water, air, feces, human nasopharynx.
-- diseases:
①pyogenic infection;
②toxin-associated disease
e.g., scarlet fever;
③hypersensitive disease;
glomerulonephritis

rheumatic fever
 Biological characteristics
• Morphology & culture properties
• G+, round, arranged in chains,
no other special structure
--hyaluronic acid capsule in the early age (2-4hr)
Nutrient requirement: high
 fastidious, “flesh-eating bacteria”
Facultative/obligate anaerobes
On blood agar:
different hemolysis
 Biological characteristics
• Classification
 Hemolytic activity
• -hemolytic strep.
Incomplete hemolysis (α)
Opportunistic pathogens
e.g., S.pneumoniae, S.viridans
-hemolytic strep.
Complete hemolysis ()
Main human pathogens
e.g., S. pyogenes
-streptococcus
No hemolysis,
No pathogenicity
rarely cause infection
 a

c
 Biological characteristics
Antigenic structure Cell wall
Polysaccharide antigen
 group-specific antigen
 20 groups (A~H, K~V)
 groups A, B, C, D, G--human diseases
 Group A--main human pathogens Streptococcus
Protein antigen
 type-specific antigen
Capsule
 M protein Protein M, R, T
 Group A: >100 serotypes Polysaccharide

Peptidoglycan

schematic picture of strep.

Biological characteristics
Resistance
 Heat labile: be killed at 60℃, 30 min
 Antibiotics sensitivity:
 penicillin, erythromycin, tetracycline, sulfonamide, etc.
 Pathogenicity (Strep. Group A)

• Virulence factors
 Surface structures
LTA: adhere to host cells
M protein:
a.Anti-phagocytosis
b.Be associated with rheumatic heart disease & glomerulonephritis
c.Adhere to epithelial cells
d.Clump platelets and leukocytes
Streptococcus pyogenes
* anti-phagocytosis
* adhere to epithelial cells
* clump platelets and leukocytes
* heat stable; acid stable (pH2)

hair-like structure
Pathogenicity (Strep. Group A)
 Invasive enzymes
Hyaluronidase (spreading factor)
---split hyaluronic acids
Streptokinase (SK)
---prevent plasma clotting/dissolve plasma clot
Streptodornase (SD)
---degrade DNA in pusreduce pus viscosity

 Toxins---exotoxins
• Streptolysin (hemolysin)
• Erythrogenic toxin (pyrogenic toxin /scarlet fever toxin)
Streptolysins

Properties Streptolysin O (SLO) Streptolysin S (SLS)

O2 oxygen-labile oxygen-stable
Chemical Protein (MW 60,000) Polypeptide (28aa)
Antigenicity Strong (ASO test) Weak
Biological destroy WBC, platelet, destroy WBC,
activity MΦ, NC virulent to many tissues

ASO: Anti-streptolysin-O, antibody

ASO test (Anti-Streptolysin O test)
Definition: a neutralization test between the toxin (SLO)
and its specific anti-toxin (ASO) which is used to diagnose
or monitor rheumatic fever caused by Group A strep.

ASO titer begins to rise from 2-3wks after primary infection

and maintain ~1 year.
ASO titer >1:250---recent infection or repeated infection
>1:400---active rheumatic fever
Erythrogenic toxin
pyrogenic toxin /scarlet fever toxin
heat stable protein
Serotype A, B, C;
 no cross-reactivity
Scarlet fever
Pathogenicity (Strep. Group A)
Diseases
 Transmission:
Respiratory tract, trauma, gastro-intestinal tract
3 types of infections
 Pyogenic infection
 Toxin-associated diseases

 Hypersensitive diseases
 Pyogenic infection

 Local purulent infections:

Acute Tonsillitis (There
pharyngitis, tonsillitis, cellulitis,
is a risk of developing
puerperal fever, erysipelas(lymphangitis) Rheumatic Fever)

impetigo; otitis, pneumonia, meningitis

Systemic infection: septicemia

Abscess with
surrounding cellulitis
Erysipelas

Erysipelas on
the cheek
Toxin-associated diseases
scarlet fever:
 scarlet fever toxin-producing strains
 Pharyngitis → scarlet fever

Strawberry tongue

paly around mouth Faint red rash

Hypersensitive disease

Acute glomerulonephritis
-edema, hypertension, hematuria and proteinuria

Rheumatic fever
-involve: heart, joints, subcutaneous tissue, and CNS;
-fever, migratory polyarthritis, carditis
Prevention and treatment
Treat the pharyngitis and tonsillitis in time
Antibiotics: penicillin for the first choice
Section 3 Pneumococcus
General properties
G+, arranged in pairs, bullet shape
capsule: polysaccharide
 >90 serotypes
 >20 serotypes cause human diseases
 only encapsulated strains are virulent
fastidious
 blood agar or chocolate blood agar
-hemolysis
autolysis
 central depression or central collapse

bile solubility test: “+”

--distinguish from other -hemolytic
streptococci (S.viridans).
ferment inulin
S. pnuemoniae capsule

chocolate blood agar

S. pnuemoniae umbilicated colonies
Note the center collapse due to autolysis
distinguish from other -hemolytic
streptococci (S.viridans).
Bile solubility test
Optochin susceptibility test
Animal experiment---mouse toxicity test
Quellung reaction (or capsular swelling reaction)
Pathogenicity
Virulence factor:
Capsule
Pneumolysin & Neuraminidase

Main disease:
lober pneumonia;
bronchitis
otitis in infants and young children;
bacteremia, septicemia;
meningitis in various age groups;
A gram-positive coccus that grows in pairs or short
chains and that is α-hemolytic, optochin-sensitive
is ( )
A. Streptococcus pyogenes
B. Streptococcus agalactiae
C. Staphylococcus aureus
D. Streptococcus pneumoniae
E. Streptococcus mutans
 Neisseria meningitidis Neisseria gonorrhoea
Biological
characteristics

Virulent factors

Diseases

Immunity
Diagnosis

Prevention and
treatment
 Section 4 Neisseria

Genus Neisseria: ~23 species and subspecies

N. Meningitidis
--meningitis
--low prevalence but
high mortality

N. Gonorrhoeae
-- human gonorrhea
-- high prevalence but low mortality
Common biological characteristics

polymorphonuclear cells
G-, coffee bean-shaped or
kidney-shaped, in pairs
capsules and pili
fastidious
 enriched medium (chocolate
blood agar plate)
Grow better in 5~10% CO2
Resistance: very low
N. Meningitidis / meningococcus
Pathogenicity
 Virulence factor:
 Pili -- attach to nasopharyngeal mucosa
 Capsule
 Endotoxin
 Fever
 capillary endothelial cell injury
 local thrombosis
 hemorrhage
 transmission: respiratory droplets
 Disease: epidemic cerebrospinal meningitis
 Symptoms: sudden onset; high fever; severe
headache; pain and stiffness of the neck, back, and
shoulders; nausea and vomiting; septicemia.
N. meningitidis (fever, chills, skin ecchymosis,
hepatosplenomegaly)
bacteremia cross the brain
nasoparynx or septicemia barrier
(asymptomatic, most
are carriers, only 2~3% meninges
go to next stage)
epidemic cerebrospinal
（2-3%） meningitis
blood stream
severe headache,
nausea, vomiting,
stiff neck,
 N. Meningitidis / meningococcus
Clinical typies:
common type: ~ 90％；
outbreak type：10-20%
high mortality: 40~60%；
chronic septicemic type：rare
N. Gonorrhoeae /gonococcus

• Virulence factors
• Pili
• IgA1 protease
• Outer membrane protein (OMP)
• LOS (endotoxin)

• Transmission:
sexual contact;
indirect contact;
(basin, towel, etc)

Diseases: Gonorrhea
Gonorrhea --- STD

Adults
 Clinical disease: Genitourinary tract infection
Male: urethritis, prostatitis; epididymitis;
Female: urethritis, cervix inflammation;
infertility
Newborns
 Ophthalmia neonatorum→blindness
Prevention and treatment
Penicillin
0.5-1% Silver nitrate-- ophthalmia neonatorum