CHAPTER 21

Pseudomonas, Burkholderia, and

Similar Organisms

Section 8 heading – Mac positive, Oxidase positive Gram

negative bacilli and coccobacilli (14th Ed.)

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 Objectives
• Describe the normal sources (habitat) for Pseudomonas aeruginosa, Burkholderia cepacia,
B. pseudomallei, and B. mallei, including the routes of transmission.
• Identify the factors that contribute to the pathogenicity of P. aeruginosa, and explain the
physiologic mechanism for each.
• List the various disease states associated with P. aeruginosa and Burkholderia sp.
• Compare and contrast the Gram-stain appearance of the gram-negative bacilli discussed
in this chapter.
• List the appropriate identification scheme for identifying P. aeruginosa.
• Describe the media and chemical principle of each media used, including differential and
selective agars that aid in the cultivation of Pseudomonas, Brevundimonas, and Ralstonia.
• Describe the potential therapies for B. cepacia and B. pseudomallei and the concerns
regarding optimal therapy.
• Describe and identify the patterns of antibiotic resistance in P. aeruginosa.

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Habitat
• Pseudomonas aeruginosa
• Is found in the environment; survives well in domestic environments.
• Is transmitted via ingestion or exposure to contaminated food, water, or
medical devices.
• Burkholderia cepacia
• Is found in the environment.
• May colonize respiratory tract of patients with cystic fibrosis.
• Is transmitted via exposure to medical devices.
• B. pseudomallei
• Is found in tropical and subtropical areas. Cause melioidosis
• Is transmitted via inhalation or direct inoculation through mucosa.
• B. mallei
• Is an agent of glanders in horses.
• Is associated with close animal contact – zoonotic diseases.
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 Pseudomonas aeruginosa
• Is an opportunistic pathogen that can cause community- or
hospital-acquired infections (CAI or HAI’s).
• Virulence factors include:
• Exotoxin A
• Endotoxins
• Proteolytic enzymes
• Alginate
• Pili
• Intrinsic resistance to many antimicrobial agents

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Infections – Pseudomonas aeruginosa

Community-Acquired (CAI’s) Hospital-Acquired (HAI’s)

• Folliculitis • Respiratory tract Luki tube
• Otitis external (Swimmers ear) • Urinary tract
• Eye infections • Wounds
• Osteomyelitis • Bacteremia
• Endocarditis in intravenous (IV) • Central nervous system
drug users
• Respiratory tract infections in
patients with cystic fibrosis

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Pathogenic, Gram-Negative, Aerobic Bacilli
• Pseudomonads
– Pseudomonas aeruginosa:
– Diseases: “Swimmer’s ear”, Eye, wounds, bacteremia, UTI, GIT etc.
– Only an opportunistic pathogen esp. burn victims or Cystic fibrosis pt’s
– Treatment: is difficult due to drug/antibiotic resistance:
– NB! Very resistant – Metabolize drugs (proton/drug antiports) & biofilm formation
– Aminoglycosides (Amikacin, Gentamycin & Tobramycin)

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 Burkholderia infections
• Includes fulminant lung infections and bacteremia.
• Cystic fibrosis
• Chronic granulomatous disease

• May result from exposure to contaminated medical solutions or

devices.
• Septicemia
• Urinary tract infections
• Respiratory tract infections

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 Gram-Stain Appearance
• Medium-size straight rods
• Brevundimonas diminuta
• Long, straight rods
• Burkholderia mallei
• Coccobacillus
• Pseudomonas pseudomallei
• Small with bi-polar staining
• The Centers for Disease Control and
Prevention (CDC) group Ic
• Thin pleomorphic rods

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 Identification – Pseudomonas aeruginosa
• Spreading, flat colony with metallic sheen Beta haemolytic,
Metallic colonies on
(Blood agar) Blood agar
• Beta-hemolytic on BA, oxidase positive;
catalase positive
• Grapelike to tortilla odor
• May also be mucoid
• Non-lactose fermenter (Mac plt)
NLF colonies on
• Grows well at 42°C MacConkey agar
• Oxidizes glucose and xylose Mucoid colonies –
in CF patients

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Pathogenic, Gram-Negative, Aerobic Bacilli
• Pseudomonads
– Pseudomonas aeruginosa:
– Microscopy – GNB, motile, encapsulated, non-sporing
– Culture: Colonies – NLF; sweet odor (Grape-like), Pyocyanin
(Blue/green); obligate aerobes; 370C for 24hrs, Oxidase Positive
– Virulence: Attachment – Fimbriae, Adhesins, Neuraminidase;
– Polysaccharide capsule;
– Toxins – Lipid A; Exotoxin A; Exoenzyme S
– Elastase (breaks elastic fibers)
– Pyocyanin (reactive O2)

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A Pseudomonas aeruginosa infection

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Ecthyma gangrenosum
Cultivation
• 5% sheep blood and chocolate agars
• Most organisms grow well on routine media. Beta hemolysis on Blood agar
• MacConkey agar
• All organisms will grow on this medium except Brevundimonas vesicularis.
• Cetrimide agar – selective isolation and presumptive ID of P. aeruginosa

• Pseudomonas cepacia (PC) agar

• B. cepacia breaks down the pyruvate, resulting in a red color.
• Oxidative-fermentative base-polymyxin B-bacitracin-lactose (OFPBL) agar
• B. cepacia ferments lactose and appears yellow.
• Ashdown medium
• Burkholderia pseudomallei will turn red on this medium.
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• Nutrient Broth: Blood agar: MH/Nutrient agar:

Cetrimide agar

MacConkey agar
Therapy
• Antimicrobial therapy for B. cepacia and B. pseudomallei rarely
eradicates B. cepacia, and therapy for melioidosis remains
controversial.
• Burkholderia cepacia
• Piperacillin • Chloramphenicol
• Ceftazidime • Trimethoprim-sulfamethoxazole
• Ciprofloxacin

• B. pseudomallei
• Ceftazidime • Amoxicillin-clavulanate
• Piperacillin-tazobactam • Imipenem
• Ticarcillin-clavulanate • Trimethoprim-sulfamethoxazole

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Therapy – Pseudomonas aeruginosa
• P. aeruginosa is intrinsically resistant to various antimicrobial agents.
• P. aeruginosa readily acquires resistance to active agents.
• Therapeutic options include: • Carbapenems:
• Antipseudomonal beta-lactam • Imipenem
with or without an aminoglycoside • Meropenem
• Aminoglycosides:
• Quinolones
• Gentamicin
• Piperacillin-tazobactam
• Tobramycin
• Ceftazidime (CAZ)
• Amikacin-netilmicin
• Cefepime (CEF)
• Fluoroquonolones:
• Aztreonam
• Ciprofloxacin
• Levofloxacin
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