Pseudomonas & Burkholderia Flashcards
What is seen here?
Pseudomonas aeruginosa
Colored by counter stain
Pseudomonas aeruginosa are Gram-______, _____, _____
negative, aerobic, rods
What can be seen in this image here? What is its purpose/function?
P. aeruginosa possess polar flagella which assist in Motility & Attachment
Single flagellum
What is seen here? What is an identifying characteristic?
P. aeruginosa – Fruity, grape-like smell
Produce metabolite called Aminoacetophenone –> responsible for grape smell
Label this image accordingly
P. aeruginosa – Diffusible pigments
P. aeruginosa is an ________ pathogen
opportunistic
can be detected in healthy animals
P. aeruginosa can be found where in the environment? What species does it effect?
(everywhere)
1. Water
2. Soil
3. Plants
4. Hospitals (nosocomial infection)
5. Home
Love moist environments and plastics
immunocompromised individuals –> devestating
Humans, dogs, cats, bovine, sheep, chickens, horses, snakes, hippos (pretty much all animals)
Why does P. aeruginosa have anti microbial resistance?
Ability to upregulate impermeability so drug can not penetrate it.
Produce beta lactamases that degrade beta lactam antibiotics such as penicilins
theey also have muktidrug efflu pumps that pump urg out of bat4eira, making them reistant to antibotiocs
AMe altes riboomoal inding sites –> ? ineffective
priduce alginate = slime like mucoid polysacchride
visoucs gel surrounding bateira assit in generating biofilm protets abcteria from antibiotics.
What are the toxins Pseudomonas possesses?
Toxins:
Exotoxin A (ADP-ribosylation of EF2),
Phospholipase C (hemolysin),
Pyocyanin (ROS),
Pyoverdine (Siderophore)
Name the structures that pseudomonas possesses that assists with attachment.
Pili, Flagella,
LPS (endotoxin) recognized by immune cells and induces strong inflammatory response –> septic shock
Name the structures that pseudomonas possesses that assists with Biofilm formation.
Mucoid EPS (Alginate)
biofilm protects bacteria from antibiotics nad host immune respoinses.
Name the type 3 secretion system that pseudomonas possesses
see slide again.
Exoenzyme S
important because enodtoxin is translocated intohost cells thorugh type 3 secretion system
induces cell death by apoptosis and prevents phagocytic killing
Exotoxin A –> endocytosis of toxin ?
Phosphoplipase C –> lyses host membrne and leads ot cell deeath
Pyocyanin –> taken up by host cell where it generates reactive oxygen speceis
pyoveridine –> binds to iron and provides essential nutrient to microbes sequestong microbes from host cell
degradative enzymes
What are the symptoms and side effects of P. aeruginosa infection ?
- Skin infections
- Ear infections
- Eye infections
- Respiratory infections
- Bloodstream infections
- Other
Skin infections caused by P. aeruginosa cause _____ ___ in sheep.
fleece rot
Water penetration of fleece allows pseudomonas infection. reported in conditions associated with prolonged or heavy rainfall.
Skin infections caused by P. aeruginosa cause ________ in dogs.
pyoderma
Ear infections caused by P. aeruginosa cause ________ ____ in dogs & cats.
otitis externa
Ear infections caused by P. aeruginosa cause ________ ____/_____ in chinchillas.
otitis media, interna
Eye infections caused by P. aeruginosa cause ________ _________ in dogs, cats, and horses.
Ulcerative keratitis
Respiratory infections caused by P. aeruginosa cause ______________ in dogs, cats, and chinchillas.
pneumonia
Bloodstream infections caused by P. aeruginosa cause ___________ & septicemia in chinchillas.
Bacteremia & Septicemia
Through bloodstream –> brain, GI tract, urinary tract, etc.
Other types of infections caused by P. aeruginosa cause _________ in cattle and sheep and _______ _____ in snakes.
mastitis, necrotic stomatitis
What specimens should you collect from a patient that you suspect is infected with P. aeruginosa?
pus, respiratory aspirates, milk samples, ear swabs, etc.
What cultures should you use to isolate the bacterium if you suspect your patient is infected with P. aeruginosa?
- Nutrient agar
- MacConkey agar
- Blood agar
incubated aerobically at 37°C for 24 to 48 hours
What biochemical tests should you use to identify the bacterium if you suspect your patient is infected with P. aeruginosa?
Oxidase, catalase
positive for both!
What molecular-based techniques should you use to confirm the diagnosis of the bacterium if you suspect your patient is infected with P. aeruginosa?
PCR and sequencing
What is seen here?
What is seen here?
What is seen here?
How would you treat your patient if they were infected by P. aeruginosa?
Identify and eliminate the sources of infection: water, diet, surroundings.
1. Antibiotic therapy
- Perform Antimicrobial susceptibility testing prior to administering antibiotics
2. Vaccines
- for farmed mink and chinchillas
Which drugs are less effective against Pseudomonas?
Penicillin, Ampicillin, Tetracycline, First- and
Second-generation Cephalosporins,
Chloramphenicol, Clindamycin, Erythromycin,
and Trimethoprim-sulfadiazine
Pseudomonas fluorescens is a gram-_______, _____, _______ _____.
negative, motile, aerobic rods
Pseudomonas fluorescens is unable to ferment _______.
glucose
Pseudomonas fluorescens produces ________ pigment (_______).
fluorescent, pyoverdine (ciderophore = high affinity iron chelating compound essential for aquistion of iron and ? growth).
Pseudomonas fluorescens is present in ____ and _____.
* Skin ulcers and septicemia in reptile and
freshwater fish
soil, water
Pseudomonas fluorescens is causes skin _____ and _______ in reptiles and
freshwater fish.
ulcers, septicemia
What is seen here?
Pseudomonas fluorescens produces a fluorescent pigment (pyoverdine)
What is seen here?
Skin infection with Pseudomonas fluorescens
in carp
Burkholderia mallei - Glanders was formerly ________ ______.
Pseudomonas mallei
Burkholderia mallei - Glanders is gram-______, ___-motile, ______ coccobacillus.
negative, non, aerobic
can replicate in phagocytic cells?
Burkholderia mallei - Glanders is a _______ ________ pathogen.
facultative, intracellular
Burkholderia mallei is the causative agent of ________ in horses, donkeys, and mules.
Glanders
Burkholderia mallei - Glanders has been reported in what countries?
Africa, Asia, the Middle East, Central/South America
Burkholderia mallei - Glanders is a Category __ ________ agent by the CDC
B, bioterrorism
What can be seen?
Burkholderia mallei - Glanders
B. mallei – Virulence factors
- Capsule
- LPS
- Multiple secretion systems
- Quorum sensing mechanisms
- Adhesion proteins and fimbriae
B. mallei causes ________ in horses and is considered a ______, ______, _______, ________ disease
glanders, contagious, sporadic, endemic, zoonotic
B. mallei – Glanders in horses is transmitted from horses to ______ or to ___-_____ species via what routes?
humans, non-equine, contact, ingestion, inhalation
Horses with Glanders are characterized by the formation of ______ and _____ on
the ____ and in the ________ tract
nodules, ulcers, skin, respiratory
The acute form of Glanders causes?
fever, septicemia, pneumonia
The chronic form of Glanders causes?
- Pulmonary form
- Cutaneous form (farcy)
- Nasal form
The pulmonary form of Glanders causes?
Tubercle-like lesion (nodules)
The cutaneous form of Glanders causes?
Nodules, ulcers, yellowish pus
The nasal form of Glanders causes?
Ulcerative nodules, Nasal discharge
What is seen here?
Glanders: Pulmonary form
Black, miliary nodules in the lung; can be calcified and spread disease to other parts of the respiratory tract
What is seen here?
Ulcers
Glanders: Cutaneous forms (Farcy)
What is seen here?
Healing ulcers and scars
Glanders: Cutaneous forms (Farcy)
What is seen here?
Ulcers
Glanders: Cutaneous form (Farcy)
What is seen here?
Glanders: Nasal form
Mucopurulent nasal discharge
What is seen here?
Glanders: Nasal form
Nodules and ulcers in the nasal conchae
What specimens should you collect from your patient if you suspect that your patient is B. mallei?
Pus, wound swabs, etc.
What media should you use to isolate B. mallei?
- Media containing 1% glycerol (no growth on MacConkey agar)
- Incubated aerobically at 37°C for 2-3 days
What tests would you use to identify B. mallei?
Identification for isolates:
Biochemical tests, PCR, real-time PCR
What serological tests would you use to identify antibodies against B. mallei?
Serological tests (identification of antibodies)
- Specimen: Blood
- Complement Fixation (CF) test (approved by the OIE)
- Lipopolysaccharide (LPS)-western blot
- Indirect enzyme-linked immunosorbent assay (I-ELISA)
-Competitive ELISA (C-ELISA)
Should be performed with a second test?
What is seen here?
B. mallei
What test has been performed here? For what reason
B. mallei – Diagnosis
The Mallein test
Left = negative
Right = positive
Similar to tuberculin skin test.
Mallein is a glycoprotein extracted from B. Mallei;,used as antigen to skin in lower eye lid or under skin in the middle of the neck. The horse is then monitored for swelling of the lower eye lid or neck.
Can use it for anticomplementary sera ; medicatio or infection?
Specimens collected from patients infected with B. mallei should be processed in ?
a biohazard cabinet
You can control B. mallei by 1. Effective ______ and ______ with a contact time of __ hours. The disinfectants you can use are?
2. ______ therapy (in _____ areas; ______ in the U.S.) using?
Unfortunately, there no ____.
cleaning, disinfection, 6
Formalin (1.5%)
Iodophor (2%)
Sodium hydrochloride
70% ethanol etc.
Antibiotic, endemic, euthanasia
Tetracyclines – Doxycycline
Sulfonamides – Sulfadiazine/Trimethoprim (TMP)
Ciprofloxacin, Streptomycin, Novobiocin, Gentamicin,
Imipenem, Ceftrazidime
vaccine
Treatment is prolonged and often ineffective.
Treatment in US is not allowed in the US; should be returned to country of origin or euthanized.
Burkholderia pseudomallei – Melioidosis
Gram- ______, _____, _____ bacillus
_______ staining –> referred to as ________ ___ appearance.
A facultative _______ pathogen
Can stay ______ for years. Naturally found in ___ and ___
The causative agent of _______.
Endemic in ______ _____ and northern _____, and _____ pacific. More common in ______ climates.
________ in many organs
Known as ______ disease or ______. Clinical ? in horses. Melliodoises caused by contact with env while glander is caused by contact with infected animals.
A Category __ bioterrorism agent by the CDC
negative, aerobic, motile, Bipolar, safety pin, intracellular, dormant, soil, water, melioidosis, southeast Asia , Australia, south, Abscesses, Abscesses, Whitmore’s, pseudoglanders, B
What is seen here?
Burkholderia pseudomallei – Melioidosis
Safety pin appearance
B. pseudomallei – Virulence factors
- Capsule
- LPS
- Multiple secretion systems
- Quorum sensing (Biofilm)
- Type IV pili
Clinical infection of B. Pseudomallei causes:
* _____ skin _____
* _____ _____ formation
* _________ formation
* Chronic ______
* Acute _______
Local, ulcers, Chronic abscess, Granuloma, pneumonia, septicemia
What are the animal hosts of B. pseudomallei ?
Sheep, goats, pigs, horses. dogs, cats, cattle, people
B. pseudomallei infection in sheep will cause?
Lung abscesses and pneumonia
B. pseudomallei infection in goats will cause?
Mastitis
B. pseudomallei infection in pigs will cause?
Asymptomatic lesions
on the spleen
B. pseudomallei infection in horses will cause?
Pseudoglanders
B. pseudomallei infection in dogs will cause?
Abscess formation of
multiple organs
B. pseudomallei infection in cats will cause?
Abscess formation of
multiple organs
B. pseudomallei infection in cows will cause?
Pneumonia, or central nervous system disease
B. pseudomallei infection in cows will cause?
Pneumonia
B. pseudomallei infection in people will cause?
Pneumonia and abscesses
Subclinical disease
What specimens would you collect from your patient if you suspected it was infected with B. pseudomallei?
Pus from abscesses, affected tissues, and blood
How would you isolate from B. pseudomallei from the samples you collected?
Grown on blood agar and MacConkey agar
Incubated aerobically at 37°C for 2-3 days
Greyish-white, non-hemolytic on blood agar
How would you identify B. pseudomallei from the samples you collected?
Identification criteria for isolates:
Colonial morphology and musty odor
PCR techniques
Serological tests:
ELISA,
Complement fixation test
Indirect hemagglutination test
What is seen here?
B. pseudomallei
Blood agar
How would you treat your patient if it was infected with B. pseudomallei?
Antibiotic therapy (resistant to many; relapses are common), surgical drainage of large abscesses, enucleation in the case of eye infection.
Antibiotic therapy for a case of B. pseudomallei: ________ plus ________/__________ or _______ for 2 weeks
followed by oral administration of _________/________, and _______, with or without _________ for 12 to 20 weeks.
Ceftazidime, Sulfonamide, trimethoprim, meropenem, sulfonamide, trimethoprim, doxycycline, chloramphenicol,
What is the risk of using antibiotic therapy to treat B. pseudomallei?
Drug resistance, relapse.
highlighted in purple = important