MICROBIOLOGY PPT LECTURE NOTES | Karya Tulis Ilmiah

(1)

(2)

(3)

Morphology & Identification

 _{Gram positive}_{Gram positive}

 _{Facultative anaerobes}_{Facultative anaerobes}  _{Grape like-clusters}_{Grape like-clusters}

 _{Catalase positive}_{Catalase positive}

 _{Major components of}_{Major components of}

normal flora

_skin_skin _nose_nose

Catalase test （过氧化氢

酶）

(4)

(5)

(6)

Pathogenesis

• Catalase

• Coagulase

• Hyaluronidase and Lipase ipase

• Hemolysin or sphingomyelinase Csphingomyelinase C

• Leukocidin

• Exfoliative Toxin

• Toxic Shock Syndrome Toxin (superantigen) • Enterotoxins Protein A Protein A immunoglobulin immunoglobulin Fc receptor Fc receptor BACTERIUM BACTERIUM PHAGOCYTE PHAGOCYTE

(7)

Pathogenesis of staphylococcal infections

Stye:

Stye:麦粒肿_麦粒肿

Carbuncle:

Carbuncle:痈痈

Impetigo

(8)

Suppurative

• _Skin_Skin

 _{Furuncle; Protein A, Leukocidin, Hemolysin}_{Furuncle; Protein A, Leukocidin, Hemolysin}  _{Stye; lipase}_{Stye; lipase}

 _{Impetigo; contagious}_{Impetigo; contagious}  _{Epidermal necrolysis}_{Epidermal necrolysis}

 _{Exfoliative Dermatitis (6,7,8); Exfoliative toxin}_{Exfoliative Dermatitis (6,7,8); Exfoliative toxin}  _Mastitis_Mastitis

 _{Abscess (deep tissue); granulation; coagulase, hyaluronidase (burn,}_{Abscess (deep tissue); granulation; coagulase, hyaluronidase (burn,}

wound)

• _Systemic_Systemic

(9)

• _{Food poisoning}

• _{Toxic shock syndrome}

• _babies

– scalded skin syndrome_{scalded skin syndrome}

* Exfoliatin_Exfoliatin

• fever_fever

• scarlatiniform_{scarlatiniform} rash rash

• desquamation_desquamation

• vomiting_vomiting

• diarrhea_diarrhea

• myalgias_myalgias • not a human infectionnot a human infection

• food contaminated from humans _{food contaminated from humans}

– growth _growth

– enterotoxinenterotoxin

• onset and recovery both occur within few hoursonset and recovery both occur within few hours

(10)

Infections associated with indwelling devices

(11)

Laboratory

• _{A. Direct examination; Gram Stain} • B. Primary media; BAP

• C. Differential Tests. 1. Mannitol Salts

2. Coagulase 3. DNase

• _{D. Phage typing}

• E. Antibiotic Sensitivity (plasmid, B lactamase) : penicillin

(12)

GRAM POSITIVE COCCI

S. aureus

 hemolytic mannitol yellow

-Staphylococcus (Clusters) Streptococcus (pairs & chains) Catalase

• BETA: Bacitracin S .pyogenes (group A) CAMP/Hippurate S. agalactiae (group B)

Hemolysis Coagulase

S. epidermidis

nonhem olytic (usua lly) mannitol

white

(2) ALPHA: Optochin/Bile Solubility S. pneumoniae

• GAMMA: Bile Esculin 6.5% NaCl Group D* Enterococcus

Bile Esc ulin 6.5% NaCl Group D* Non-Enterococcus

(*can also be beta or alpha hemolytic)

Note: Strep. viridans

are alpha hemolytic and negat ive for all the tests below + + + + + + +

(13)

Staphylococcus epidermidis

• major component skin flora_{major component skin flora}

• opportunistic infections _{opportunistic infections}

– less common than less common than S.aureusS.aureus

• nosocomial infectionsnosocomial infections

– heart valves_{heart valves}

• Identification_{Identification}

– Non-hemolytic _{Non-hemolytic}((sheep blood agarsheep blood agar))

– Does not ferment mannitol_{Does not ferment mannitol}

– Non-pigmented _{Non-pigmented}

– _{Coagulase-negative}_{Coagulase-negative}

Staphylococcus saprophyticus

• _{urinary tract infections}

• _{coagulase-negative}

(14)

(15)

(16)

(17)

(18)

Morphology & Identification

• _{facultative anaerobe}

• _{Gram-positive}

• _Chains

_{or pairs}

• _Catalase

_negative

(19)

Cell surface structure of S pyogenes and extracellular substances

•Lancefield groups_{Lancefield groups}

*one or more species per group_{one or more species per group}

*surface antigens: M, T, R_{surface antigens: M, T, R}

groupable streptococcistreptococci

•A, B and DA, B and D

–most important_{most important}

•C, G, F _{C, G, F}

–Rare_Rare

Non-groupable

•S. pneumoniae_{S. pneumoniae}

–pneumonia _pneumonia

•viridans streptococciviridans streptococci

–e.g._e.g. S. mutans S. mutans

*dental _dental

caries

(20)

Lipoteichoic Acid and F-protein

fibronectin

lipoteichoic acid

F-protein

epithelial cells

(21)

M protein

• _{major target}

– _{natural immunity}_{natural immunity}

• _{strain variation}

– antigenicity_antigenicity

• _re-infection

(22)

M protein

M protein M protein fibrinogen fibrinogen r r r peptidoglycan peptidoglycan r r r IgG IgG Complement

Complement IMMUNEIMMUNE

NON-IMMUNE

(23)

(24)

Hemolysis

alpha

beta

(25)

(26)

(27)

S

.

_.

pyogenes (

Group A)

-suppurative

• _{affect all ages peak incidence at}_{affect all ages peak incidence at}

5-15 years of age

• _non-invasive_non-invasive

– _pharyngitis_pharyngitis

– skin infection, impetigo_{skin infection, impetigo}

• _{invasive bacteremia}_{invasive bacteremia}

– _{toxic shock-like syndrome}_{toxic shock-like syndrome} – _{"flesh eating" bacteria}_{"flesh eating" bacteria}

(28)

• Scarlet fever _{Scarlet fever}  _rash_rash

 _{erythrogenic toxin}_{erythrogenic toxin}

• rheumatic fever _{rheumatic fever}

 _{inflammatory disease}_{inflammatory disease}  _{life threatening}_{life threatening}

 _{chronic sequalae}_{chronic sequalae}  _fever_fever

 _Heart_Heart  _Joints_Joints

 _{rheumatic NOT rheumatoid arthritis}_{rheumatic NOT rheumatoid arthritis}

• Acute glomerulonephritis _{Acute glomerulonephritis}

immune complex disease of kidney

Rheumatic fever -etiology Rheumatic fever -etiology

 _{M protein}_{M protein}

– cross-reacts heart myosin _{cross-reacts heart myosin}

– autoimmunity_autoimmunity

 _{cell wall antigens}_{cell wall antigens}

– poorly digested poorly digested in vivoin vivo

– persist indefinitely_{persist indefinitely}

Post-infectious diagnosis (serology)

• antibodies to streptolysin O_{antibodies to streptolysin O}

• important if delayed clinical sequelae _{important if delayed clinical sequelae}

occur

• superantigen_superantigen

• T cell mitogen _{T cell mitogen}

• activates immune _{activates immune}

system

(29)

Group B streptococcus -

identification

• _{neonatal meningitis}_{neonatal meningitis}

• _septicemia_septicemia • transmission _transmission

– vaginal flora_{vaginal flora}

 __ _hemolysis_hemolysis

• _{hippurate hydrolysis}_{hippurate hydrolysis} • _{CAMP reaction}_{CAMP reaction}

(30)

Group D streptococcus

• Growth on bile esculin agar_{Growth on bile esculin agar}

– black precipitate _{black precipitate}

• 6.5% saline_{6.5% saline}

• _grow_grow

– _enterococci_enterococci • no growth _{no growth}

(31)

Enterococci

• _{distantly related to other streptococci}_{distantly related to other streptococci} • _genus_genus_Enterococcus_Enterococcus

• gut flora_{gut flora}

– _{urinary tract infection}_{urinary tract infection}

• fecal contamination_{fecal contamination}

– _{opportunistic infections}_{opportunistic infections}

• _{particularly endocarditis}_{particularly endocarditis}

• _{most common}_{most common}_{E. (S.) faecalis}_{E. (S.) faecalis}

• resistant to many antibiotics _{resistant to many antibiotics}

– including vancomycin_{including vancomycin}

(32)

Viridans streptococci

• _{diverse species}_{diverse species}

• _oral_oral

• dental caries_{dental caries}

 __ _{hemolytic and negative for other tests}_{hemolytic and negative for other tests}

• _{non-groupable.}_{non-groupable.}

• includes _includesS. mutansS. mutans

– endocarditis _endocarditis

(33)

Streptococcus pneumoniae

S. pneumoniae - diplococci

• capsule: _capsule:

• pneumolysin_pneumolysin: :

• _{Surface protein adhesinand}_{Surface protein adhesinand} secretory IgA protease.

secretory IgA protease.

• Teichoic acid and the _{Teichoic acid and the}

Peptidoglycan fragment,

phosphorylchorine .

• leading cause pneumonia_{leading cause pneumonia}

– particularly young and old_{particularly young and old}

– after damage to upper _{after damage to upper} respiratory tract

respiratory tract

*e.g. following viral infection *e.g. following viral infection

• bacteremia_bacteremia

• meningitis_meningitis

(34)

GRAM POSITIVE COCCI

S. aureus

 hemolytic mannitol yellow

-Staphylococcus (Clusters) Streptococcus (pairs & chains) Catalase

• BETA: Bacitracin S .pyogenes (group A) CAMP/Hippurate S. agalactiae (group B)

Hemolysis Coagulase

S. epidermidis

nonhem olytic (usua lly) mannitol

white

(2) ALPHA: Optochin/Bile Solubility S. pneumoniae

• GAMMA: Bile Esculin 6.5% NaCl Group D* Enterococcus

Bile Esc ulin 6.5% NaCl Group D* Non-Enterococcus

(*can also be beta or alpha hemolytic)

Note: Strep. viridans

are alpha hemolytic and negat ive for all the tests below + + + + + + +

(35)

Streptex antiserum

Latex agglutination - streptococci

Quellung reaction Quellung reaction

• using antisera using antisera

• capsule "fixed" _{capsule "fixed"}

• visible microscopically_{visible microscopically}

Not optochin sensitive optochin sensitive

(36)

Prevention and Treatment

• _{Immunity ; 14 capsule types mixed}

vaccine

• _M

_{ost strains susceptible to}

penicillin

, but

resistance is

common

(37)

•

Gram negative _{Gram negative}

• _{diplococci (pairs of}_{diplococci (pairs of}

cocci)

• _{oxidase positive}_{oxidase positive}

• Culture: 5-10% CO_{Culture: 5-10% CO}₂₂

• Thayer Martin. _{Thayer Martin.}

– _selective_selective

– chocolate agar_{chocolate agar}

* _{heated blood}_{heated blood}

Neisseria

(38)

Capsule

LPS

N. meningitidis

Virulence Factors

Similar, but – Differences in utilization

Hemolysin IgA protease PILI

Opacity (OPA) proteins Outer Membrane Proteins

N. gonorrhoeae

LPS

PILI

Opacity (OPA) proteins Outer Membrane Proteins IgA protease

NO capsule NO hemolysin

X

(39)

Neisseria gonorrhoeae

Using the Gram stain in patient specimens, the organisms are most often observed in polymorphonuclear leukocytes

Gram stain of pure culture Urethral exudate

• _{After 2-14 days}_{After 2-14 days}

•F_Found only in manound only in man

• _G_G_onorrhea_onorrhea_:_:_{second most common venereal}_{second most common venereal}

disease

(40)

Pili = key in anchorage of organisms

to mucosal epithelium.

Nonpiliated gonococci are avirulent

Porin proteins (Por) = prevent phagolysosome fusion & allow intracellular survival [ also called protein I]

Opacity proteins (Opa) = binding of organisms to epithelium [also called protein II]

Reduction-modifiable proteins (Rmp) = protection against bactericidal antibodies [ also called protein III]

Neisseria gonorrhoeae

(41)

Bartholin’s Duct

(42)

Disseminated gonococcal infection (DGI).

Fever, polyarthritis

(or monoarticular septic arthritis), and/or dermatitis

(pustules on a hemorrhagic base).

Purulent conjunctivitis/Ophthalmia neonatorum Infection in newborns during vaginal delivery

(43)

Smear

• polymorphonuclear cell_{polymorphonuclear cell}

• Gram negative cocci_{Gram negative cocci}

many in cellsmany in cells

• Culture_Culture

• 

_

lactamase-resistant cephalosporin

_{lactamase-resistant cephalosporin}

–

_{e.g. ceftriaxone}

• _{resistant strains}

–

_common

–

_produce



_

lactamases

_lactamases

–

_{destroy penicillin}

Antibiotic therapy

(44)

N. meningitidis

(the "meningococcus")

(45)

Neisseria

meningitidis

• _resides_resides_{in man only}_{in man only}

• usually sporadic cases _{usually sporadic cases}

– mostly young _{mostly young}

children

• outbreaks _outbreaks

– _adults_adults

– crowded conditions _{crowded conditions}

* _{e.g. army}_{e.g. army}

barracks

• _1-4_1-4_days_days

• _S_S_{econd most common}_{econd most common}

meningitis

– _{pneumococcus,}_{pneumococcus,}

most common

• _F_F_{atal if untreated}_{atal if untreated} • _R_R_{esponds well to}_{esponds well to}

antibiotic therapy

antibiotic therapy – penicillin penicillin

Upper respiratory

tract

tract infection infection

– _{adhesion pili}_{adhesion pili}

Bloodstream

Brain

Meningococcal

meninigitis

(46)

Diagnosis

• _{spinal fluid}_{spinal fluid}

– Gram negative Gram negative diplococci

diplococci

within within

polymorphonuclear cells

– _{meningococcal}_{meningococcal}

antigens

• _Culture_Culture

– Thayer Martin agarThayer Martin agar

• capsule_capsule

– _{inhibit phagocytosis}_{inhibit phagocytosis}

• anti-capsular antibodies_{anti-capsular antibodies}

– stop infection_{stop infection}

• _{antigenic variation}_{antigenic variation} – _serogroups_serogroups

• vaccine _vaccine

– multiple serogroups _{multiple serogroups}

Prevention

(1)

Bartholin’s Duct

(2)

Disseminated gonococcal infection (DGI).

Fever, polyarthritis

(or monoarticular septic arthritis), and/or dermatitis

(pustules on a hemorrhagic base). Purulent conjunctivitis/Ophthalmia neonatorum Infection in

(3)

Smear

• polymorphonuclear cell_{polymorphonuclear cell}

• Gram negative cocci_{Gram negative cocci}

many in cellsmany in cells

• Culture_Culture

• 

_

lactamase-resistant cephalosporin

_{lactamase-resistant cephalosporin}

–

_{e.g. ceftriaxone}

• _{resistant strains}

–

_common

–

_produce



_

lactamases

_lactamases

–

_{destroy penicillin}

Antibiotic therapy

(4)

N. meningitidis

(the "meningococcus")

(5)

Neisseria

meningitidis

• _resides_resides_{in man only}_{in man only}

• usually sporadic cases _{usually sporadic cases}

– mostly young _{mostly young}

children

• outbreaks _outbreaks

– _adults_adults

– crowded conditions _{crowded conditions}

* _{e.g. army}_{e.g. army}

barracks

• _1-4

_days

• _S

_{econd most common}

meningitis

–

_{pneumococcus,}

most common

• _F

_{atal if untreated}

• _R

_{esponds well to}

antibiotic therapy

–

penicillin

Upper respiratory

tract

infection

–

_{adhesion pili}

Bloodstream

Brain

Meningococcal

meninigitis

(6)

Diagnosis

• _{spinal fluid}_{spinal fluid}

– Gram negative Gram negative diplococci

diplococci

within within

polymorphonuclear cells

– _{meningococcal}_{meningococcal}

antigens

• _Culture_Culture

– Thayer Martin agarThayer Martin agar

• capsule_capsule

– _{inhibit phagocytosis}_{inhibit phagocytosis} • anti-capsular antibodies_{anti-capsular antibodies}

– stop infection_{stop infection}

• _{antigenic variation}_{antigenic variation} – _serogroups_serogroups

• vaccine _vaccine

– multiple serogroups _{multiple serogroups}