Pseudomonas & Burkholderia Flashcards by Meagan Murray

What is seen here?

Pseudomonas aeruginosa
Colored by counter stain

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Pseudomonas aeruginosa are Gram-______, _____, _____

negative, aerobic, rods

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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

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What is seen here? What is an identifying characteristic?

P. aeruginosa – Fruity, grape-like smell
Produce metabolite called Aminoacetophenone –> responsible for grape smell

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Label this image accordingly

P. aeruginosa – Diffusible pigments

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P. aeruginosa is an ________ pathogen

opportunistic
can be detected in healthy animals

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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)

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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.

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What are the toxins Pseudomonas possesses?

Toxins:
Exotoxin A (ADP-ribosylation of EF2),
Phospholipase C (hemolysin),
Pyocyanin (ROS),
Pyoverdine (Siderophore)

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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

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Name the structures that pseudomonas possesses that assists with Biofilm formation.

Mucoid EPS (Alginate)
biofilm protects bacteria from antibiotics nad host immune respoinses.

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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

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What are the symptoms and side effects of P. aeruginosa infection ?

Skin infections
Ear infections
Eye infections
Respiratory infections
Bloodstream infections
Other

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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.

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Skin infections caused by P. aeruginosa cause ________ in dogs.

pyoderma

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Ear infections caused by P. aeruginosa cause ________ ____ in dogs & cats.

otitis externa

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Ear infections caused by P. aeruginosa cause ________ ____/_____ in chinchillas.

otitis media, interna

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Eye infections caused by P. aeruginosa cause ________ _________ in dogs, cats, and horses.

Ulcerative keratitis

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Respiratory infections caused by P. aeruginosa cause ______________ in dogs, cats, and chinchillas.

pneumonia

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Bloodstream infections caused by P. aeruginosa cause ___________ & septicemia in chinchillas.

Bacteremia & Septicemia
Through bloodstream –> brain, GI tract, urinary tract, etc.

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Other types of infections caused by P. aeruginosa cause _________ in cattle and sheep and _______ _____ in snakes.

mastitis, necrotic stomatitis

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What specimens should you collect from a patient that you suspect is infected with P. aeruginosa?

pus, respiratory aspirates, milk samples, ear swabs, etc.

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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

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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?

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

1. Capsule 2. LPS 3. Multiple secretion systems 4. Quorum sensing mechanisms 5. 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?

1. Pulmonary form 2. Cutaneous form (farcy) 3. 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?

1. Media containing 1% glycerol (no growth on MacConkey agar) 2. 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

1. Capsule 2. LPS 3. Multiple secretion systems 4. Quorum sensing (Biofilm) 5. 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