pathogenic anaerobe gram positive bls 206
TRANSCRIPT
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Corynebacterium
Erysipelothrix
&
Listeria
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Pathogenic Anaerobic
Gram-Positive Bacilli
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1. Corynebacteria (Genus Corynebacterium)
Aerobic or facultatively anaerobic
Small, pleomorphic (club-shaped), gram-positive
bacilli that appear in short chains (“V” or “Y”
configurations) or in clumps resembling “Chinese
letters”
Cells contain metachromatic granules (visualize
with methylene blue stain)
Lipid-rich cell wall contains meso-
diaminopimelic acid, arabino-galactan polymers,
and short-chain mycolic acids
Lysogenic bacteriophage encodes for potent
exotoxin in virulent strains
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Distinguishing Features of CMN Group
Corynebacteriu
m
Mycobacterium Nocardia
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Pathogenic Corynebacterial Species
Corynebacterium diphtheriae
Corynebacterium jeikeium
Corynebacterium urealyticum
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Corynebacterium urealyticum
Urinary tract infections (UTI’s); rare but
important
Urease hydrolyzes urea; release of NH4+,
increase in pH, alkaline urine, renal stones
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Corynebacterium diphtheriae
Respiratory diphtheria (pseudomembrane on
pharynx) and cutaneous diphtheria
Prototype A-B exotoxin acts systemically
• Toxoid in DPT and TD vaccines
Diphtheria toxin encoded by tox gene introduced
by lysogenic bacteriophage (prophage)
Selective media: cysteine-tellurite; serum
tellurite; Loeffler’s
Gravis, intermedius, and mitis colonial morphology
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Epidemiology of
Diphtheria
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Virulence Factors in Corynebacterium Species
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Diphtheria tox Gene in
Beta Bacteriophage
and Prophage
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See Handout on Exotoxins
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Mechanism of Action of Diphtheria Toxin:
Inhibition of Protein Synthesis
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Molecular Structure of Diphtheria Toxin
Catalytic Region
Receptor-Binding Region
Translocation Region
A Subunit
B Subunit
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Heparin-binding epidermal growth factor on heart & nerve surfaces
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Diagnostic Schick Skin Test
TOXIN TOXOID
Immune Status to C. diphtheriae and
Sensitivity to Diphtheria Toxoid
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In vivo Detection
of Diphtheria Exotoxin
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2. Genus Listeria
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2.Listeria monocytogenes
• Small, Gram +, nonsporing rod
• End-over-end tumbling motility when grown at
20-25°C, not at 37°C
• Facultative anaerobe, ß-hemolytic
• CAMP Test positive (like Group B Streptococcus)
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Listeriosis
• Humans, domestic animals
described in ≥ 40 species of animals
usually follows ingestion
outbreaks, sporadic cases related to food
asymptomatic fecal carriage common,
especially for those in contact with domestic
animals
Incidence increases in summer, when
outbreaks of food-borne disease are more
common.
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Neonates, elderly & immunocompromised
Granulomatosis infantiseptica
• Transmitted to fetus transplacentally
• Early septicemic form: 1-5 days post-partum
• Delayed meningitic form: 10-20 days following birth
Intracellular pathogen
• Cell-mediated and humoral immunity develop
• Only cell-mediated immunity is protective
Listeriosis
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Intestinal tract of mammals & birds (especially chickens)
Persists in soil
Soft cheeses & unwashed raw vegetables
Raw or undercooked food of animal origin
Luncheon meats
Hot dogs
Large scale food recalls have become common
Distribution of Listeria?
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Natural
Reservoirs
Common Routes for
Human Exposure
Population at
Greatest Risk
Epidemiology of Listeria Infections
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Virulence Factors and Pathogenesis -
Motility
• Actin-mediated motility (ActA)
host cell actin polymerized
growth of tail by actin polymerization at end of
bacterium propels it through cytoplasm
• Uptake: induced phagocytosis
internalin
similar to M protein of S. pyogenes
(antiphagocytic), dissimilar functions
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Virulence Factors and Pathogenesis
• After entry to epithelial
cells
escapes phagosome,
multiplies in cytoplasm
exocytosis from epithelial
cell followed by
phagocytosis by MØ, PMN
multiplication followed by
death of phagocytes,
secondary phagocytosis
systemic spread
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Virulence Factors and Pathogenesis -
Listeriolysin
• Major virulence factor: listeriolysin
thiol-activated cytolysin, hemolysin
mediates escape from phagocytic vesicle
• LLO mutants: LD50 5 logs higher than WT,
do not survive in MØ
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Virulence Factors and Pathogenesis
• Bacteria encountering plasma
membrane continue to move
forward
produce protrusions extending
into adjacent cell: listeriopods
escape listeriopod in double-
membrane vesicle, enter
cytoplasm of adjacent cell
• Mediated by phospholipase
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Virulence Factors and Pathogenesis -
Actin-based Motility
• Bacteria in cytoplasm
polymerize actin, form
tails
hollow mesh forms on
surface, left behind as
bacterium moves forward
invade adjacent cells
• Actin nucleating factor: ActA
ActA localized at one end of the
bacterium, not found in tail
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Listeriolysin O?
Macrophag
e
Macrophag
e
Phagocytosis
Intracellula
r
Replication
Actin
Filaments
Intracellular Survival & Replication of Listeria
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Immune Response
• In infected mice, bacteria first appear in MØ, then
invade hepatocytes
most replication probably occurs in liver
infection of MØ leads to presentation of antigens with
MHC class I, stimulating cytotoxic T cell response
cytotoxic T cells (and NK cells) kill infected hepatocytes
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bacteria released from lysed host cells killed
bv activated MØ
T cell-deficient mice survive infection:
cytotoxic T-cell response helps clear
hepatocytes, not essential
increased susceptibility in mice unable to
produce IFN-: suggests importance of
activated MØ
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Erysipelothrix rhusopathiae
Gram-positive non-motile bacillus; forms filaments
Occupational disease of meat and fish handlers,
hunters, veterinarians
Preventable with protective gloves & clothing
Erysipeloid in humans; erysipelas in swine & turkeys
Organisms enter through break in skin
Nonsuppurative, self-limiting skin lesions with erythema
and eruption
Peripheral spread may lead to generalized infection,
septicemia and/or endocarditis
Organisms can be isolated from skin biopsy
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Epidemiology of
Listeriosis
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Natural
Reservoirs
Common Routes for
Human Exposure
Population at
Greatest Risk
Epidemiology of Listeria Infections
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Neonates, elderly & immunocompromised
Granulomatosis infantiseptica
• Transmitted to fetus transplacentally
• Early septicemic form: 1-5 days post-partum
• Delayed meningitic form: 10-20 days following birth
Intracellular pathogen
• Cell-mediated and humoral immunity develop
• Only cell-mediated immunity is protective
Listeriosis
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Methods That Circumvent Phagocytic
Killing
See Chpt. 19
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Listeriolysin O?
Macrophage
Macrophage
Phagocytosis
Intracellular
ReplicationActin
Filaments
Intracellular Survival & Replication of Listeria
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3. Erysipelothrix rhusopathiae
Gram-positive non-motile bacillus; forms filaments
Occupational disease of meat and fish handlers,
hunters, veterinarians
Preventable with protective gloves & clothing
Erysipeloid in humans; erysipelas in swine & turkeys
Organisms enter through break in skin
Nonsuppurative, self-limiting skin lesions with erythema
and eruption
Peripheral spread may lead to generalized infection,
septicemia and/or endocarditis
Organisms can be isolated from skin biopsy
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Epidemiology of
Erysipelothrix
Infection