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Genghis Khan of the Microbial World
Kills
Psuedomonas
Burned patients, cancer patients, children with cystic fibrosis (it is an opportunistic infection)
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Does Pseudomonas have flagella? Os it motile?
One flagella, motile
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Pseudomonas:
1) Gram-P or Gram-N?
2) Shape?
3) Oxidase-P or Oxidase-N?
4) Fermenter?
5) Aerobic/Anaerobic
- 1) Gram-N
- 2) Bacillus
- 3) Oxidase-P
- 4) Non-fermenter
- 5) Facultative Anaerobe: Can aspire anaerobically using nitrates
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Which bacteria produces a thick slim layer?
Pseudomonas
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What temperatures does Pseudomonas grow at?
37 and 42 C, but not 4C
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What bacteria is resistant to certain disinfectants? What disinfectants should you use?
- Pseudomonas
- Use phenols or gluteraldehydes
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Which bacteria grows anaerobically, but must have nitrates? (So it won't grow anaerobically in a glucose only tube.) Is this organism Oxidase-P or Oxidase-N?
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TSI: Pseudomonas
Red butt (no nitrates), apple red slant; pseudomonas is NOT a fermenter
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Which bacteria has a fruit odor?
Pseudomonas
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Name 3 pigments Pseudomonas produces and what they do.
- 1. Fluorescein (chelates iron)- green/yellow pigment
- 2. Pyocyanin (converts O2 to O2-)- blue
- 3. Red and black pigments also possible
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What type of pili do Pseudomonas have?
Type IV Pili
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Which of these bacteria have a Type III SS?
Pseudomonas
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How does the LPS from Pseudomonas compare with other LPS? What can it bind to?
- A little weaker, but can still cause detrimental effects.
- Binds to lectin on some mammalian cells.
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They Type IV Pili of Pseudomonas has __________ (like GC and Vibrio); the pili and flagella bind to _____ and trigger _____ production.
- N-methyl-phenylalanine
- asialo-GM1
- IL-8
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Pseudomonas contains Neuraminidase. What does this enzyme do?
GM1 --> asialo-GM1 for binding sites
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1) What exotoxin does Pseudomonas produce?
2) What activates this cell?
3) Ribosylates _____ , halting _____ and causing ____
4) Does high iron or low iron favor toxin production?
- 1) Exotoxin A
- 2) Activated by furin (tissue protease) on entry into host cell- activated by cleavage
- 3) Ribosylates EF2, halting protein synthesis and causing cell death
- 4) Low iron favors toxin production
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Name the two types of Exotoxin A produced by Pseudomonas.
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Mechanism of Action:
Exo S + Exo T + Exo Y
- Intracellular growth then apoptosis
- (Invasive)
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Mechanism of Action:
Exo U + Exo T
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Which type of Pseudomonas exotoxin A is cytotoxic?
Exo U
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Which type of Pseudomonas exotoxin A is invasive?
Exo S
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Type III Secretion Systems of P. aeruginosa
Causes cytoskeletal disruptions and ribosylates many proteins causing less DNA replication and apoptosis.
Exo S
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Type III Secretion Systems of P. aeruginosa:
Causes cytoskeletal disruptions and ribosylates multiple proteins causing less cell adhesion (blocks macrophage phagocytosis)
Exo T
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Type 3 SS of P. aeruginosa:
Which two are GTPases?
Exo S and Exo T
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Type 3 SS of P. aeruginosa:
Phospholipase that quickly kills cells when injected into the cell, strain is virulent.
Exo U
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Type 3 SS of P. aeruginosa:
An adenyl cyclase activity that disrupts actin and limits phagocytosis; favors toxin production.
Exo Y
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Which two strains of P. aeruginosa are uncommon among cystic fibrosis patients?
Cytotoxic strains: Exo U and Exo T
Good Slide w/ flow chart #25
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2 Important Pseudomonas Proteases.
- PASP: collagenase
- LEP: induces inflamation
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Pseudomonas protease that is capable of destroying host defense molecules and elastin of blood vessels (ecthyma gangrenosum)
Elastase B
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Pseudomonase protease that is present in almost all strains. It is able to destroy host defenses; may correlate with virulence.
Protease IV
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Pseudomonas Proteases:
1) Destroy host defense molecules
2) Destroy ___ (aids bacterial spreading)
3) Activate latent host ________ whose activities contribute to tissue damage
4) Cleave and activate receptors on cells that lead to _________
- 3) MMP (matrix metalloproteinases)
- 4) Pro-inflammatory cytokines (inflmmation)
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Body fluids contain _____ molecules with lectin-like properties that bind to LPS. These molecules clump and opsonize bacteria.
Surfactant
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Pseudomonas aergunosa's Regulation of Virulence
Exo Y -> cAMP -> vfr gene -> lasR gene -> Las R protein -> las I gene -> Las I protein -> Homoserine lactone -> goes back to las R gene (autostimulation)
Homoserine lactone is involved in expression of many proteins involved in virulence
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Regulation of Toxic Exoproteins:
Staph aureus: _____
P. aeruginosa:---
When do they bind/poison?
- SA= octopeptide buildup
- Pa= homoserine lactone build-up
Bind early, poison late (bind -> replicate -> poison -> spread)
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Once Pseudomonas is in the bloodstream, it activates?
complement
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Once Pseudomonas builds-up, what host defenses do the proteases destroy?
- Complement, Ab = opsonization
- Cytokines = phagocytic killing
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Most promising protein for a Pseudomonas vaccine?
This protein induces ___ response which results in a strong chemotactic event that attracts PMN to kill the bacteria.
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Which protein binds to host cells Type II SS and prevents bacteria's protein transport; thereby, stopping infections? Also a good vaccine candidate.
PerV protein
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Which protein helps form por for protein entry into mammalian cells? It may be good to use in conjucntion with PerV for P. aeruginosa vaccine.
PopB
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Multiple infections of Pseudomonas in hospital unit can indicate need for strain _____________. The best method for this is a new tool, ______, that sequencse the entire genome of the bacterial isolate.
- source identification
- rapid sequencing systems
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Pseudomonas is difficult to treat because it has intrinsic or extrinsic resistance systems.
Both!
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Pseudomonas has intrinsic resistance (via B-lactamases) to?
Penicillin G/V, ampicillin, amoxicillin, first generation sephalosproin, sulfa-trimethoprim, furantoin.
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Pseudomonas does not have intrinsic antibiotic resistance to?
Carbenapems, Gentamycin
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In what direction do genes flow between Pseudomonas and Enterobacteriaceae?
They can flow in either direction.
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Pseudomonas treatment
- 1) Beta-Lactam + Aminoglycoside
- Tircarcillin + tobramycin
- Timentin + tobramycin
- Piperacillin + tobramycin
- Ceftazidime + tobramycin
- Cefepime + amikacin
- 2) Fluoroquinolone (can be oral)
- Ciprofloxacin
- Levofloxacin
- Moxifloxacin or Gatifloxacin
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Pseudomonas aeruginosa causes what 3 outpatient infections?
- Corneal Infections (keratitis)- usually contact lens associated
- Otitis Externa- Swimmer's Ear
- Folliculitis- Hot tub related infection
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What are the two leading causes of Pseudomonas aeruginosa infection?
Burns, pneumonia
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What percentage of hospital patients are colonized with P. aeruginosa? What percentage of all nosocomial infections is this?
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Major pseudomonas infections that does not lead to bacteremia (all others lead to bacteremia)
CF lung (chronic infection)
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Chronic pseudomonas in the CF lung is in what form?
Mucoid
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Pseudomonas aeruginosa: Cystic fibrosis
What 4 virulence factors aid in survival of trachea/lung?
- Lipase
- Pyocyanin
- Proteases
- Hemolysin
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Pseudomonas aeruginosa: Cystic fibrosis
What 3 components of the organism bind to the epithelium?
LPS, pili, flagella
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Pseudomonas aeruginosa: Cystic fibrosis
Mutant bacterium starts alginate production in high/low salt environment?
high (only CF patients have high salt concentration)
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Name 3 other species of Pseudomonas that cause infections.
- P. fluorescens
- P. stutzeri
- P. putida
Treat with same antibiotics as for P. aeruginosa
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Where is Burkholderia pseudomallei found?
How is it transmitted?
What does it cause?
- Found in dry areas of SE Asia & Australia.
- Transmitted by dust (inhalation or wound infection)
- Causes multiple skin/soft tissue abscesses
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Burkholderia pseudomallei:
Do bacteria remain viable in PMN and macropahges?
Relapses after dormancy are common.
How do you treat it?
- Yes
- Treat w/ anti-pseudomonas antibiotics
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Burkholderia cepacia complex (Bcc):
1) Consists of ___ members, which are all _____ organisms
10, environmental
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Most important component of BCC
B. cenocepacia
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One of the scariest things about BCC is that it can ______
spread among CF patients, more lethal than P. aergunosa in CF patients
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Burkholderia can produce very large pili called_____. These pili bind to _____ and ____
- cable pili
- Bind to respiratory cells and protease
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Agent of antibiotic-resistant pneumonia in CA or nosocomial infections, can colonize catheters.
Stenotrophomonas maltophilia
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How do you treat Stenotrophomonas maltophilia?
Timentin or sulfa-trimethoprim
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