PSEUDOMONAS

Microbiology

 Term Pseudo = “false”; monas = “single unit”

 Term ‘aeruginosa” stems from green-blue hue
 Pseudomonas are classified as strict aerobes but some
exceptions:
 May use nitrate

 Biofilm formation
 Family Pseudomonadaceae
 Aerobic, non-spore forming Gram negative
straight or slightly curved rod (1 to 3 um in
length), polar flagella
 Non-fermenters
 Catalase and oxidase positive
 Production of pigments:
 Term ‘aeruginosa” stems from green-blue hue
Gram-stained Pseudomonas aeruginosa
bacteria (pink-red rods).
 Species
 Pseuomonas aeruginosa
 Stenotrophomonas maltophila
 Burkholderia cepacia
 Burkholderia mallei
 Burkholderia pseudomallei
Epidemiology:
The primary habitat of P. aeruginosa is
the environment, chiefly found in soil
and water.
Also found in 10% people as normal
flora of colon.
 Pseudomonas aeruginosa
 Also known as burkholderia aeruginosa.
 Other species of pseudomonas are
1. Pseudomonas cepacia
2. Pseudomonas maltophilia
3. Pseudomonas pseudomallei
 Properties:

1. Pseudomonas is a gram negative rod

with polar flagella
2. It is strict or obligate aerobe unlike the
members of enterobacteriacea.
Therefore it gets its energy through
aerobic processes and not through
fermentation.
3. It is lactose non fermenter
 P. aeruginosa and cepacia have strong ability to
withstand the disinfectants. This property also makes
them commonest nosocomial organism.
8. Pseudomonas produces two pigments.
a. Pyocanin. It gives a blue colour to the pus in
wounds.
b. Pyoverdin.( Flourescein). A yellow green pigment
which flouresces under ultraviolet light.
 Infections have resulted from the growth of
Pseudomonas in medications, contact lens solutions,
and even in some disinfectants.
 It is responsible for 10-20% nosocomial infections.
 Commonest cause of hospital acquired gram
negative pneumonia.
 Pigment Production
 Pyocyanin
 Bluish green phenazine pigment
 Soluble in chloroform and water
 Not produced by other species
 Pyoverdin(fluorescin)
 It is a greenish yellow pigment
 Insoluble in chloroform but soluble in water
 Produced by many other species
 In lab these pigments impart agar a blue
green colour
 P. aerugenosa synthesizes both pigments
while others produce only pyoverdin.
The soluble blue pigment pyocyanin is produced by
many, but not all, strains of Pseudomonas
Pyocyanin Pyoverdin
 Cultural Charecteristics

 Obligateaerobe
 Wide range of temperature 5°c-42°c

optimum 37°c
 Ordinary media –

 large,opaque,irregular,with distinctive
musty,mawkish,earthy smell
 Cont..
 Nutrient agar-
 Colonies are smooth,large,translucent,low
convex,2-4mm in diameter.
 Produce sweetish aromatic odour

 Greenish blue pigment diffuses

Pseudomonas aeruginosa colonies on agar
 Cont..
 Blood agar
 Similar to nutrient agar

 Many are haemolytic

 Mac conkey agar

 Colourless,non lactose fermenters

 Cetrimide agar

 selective media
P. aeruginosa on blood agar
 P. aeruginosa isolates may produce three types of
colony.
1. Natural isolates from soil or water produce a small,
rough colony.
2. Clinical samples produce a colony with fried-egg
appearance.
4. Oxidase positive because contains high levels of
cytochrome oxidase.
5. It can grow even in tap water with little amount of
nutrients.
6. It can survive and multiply over a wide temperature
range (20 to 42C) in almost any environment,
including one with a high salt content.
 Non-Lactose
Lactose Fermenter Fermenter

Lactose
Fermenter

MacConkey Agar
 Toxins
a. Exotoxin A
b. Exotoxin S or Exoenzyme S
Exotoxin A is the best known. It produces tissue
necrosis. Impairs protein synthesis of the cells.
Exoenzyme S is produced by bacteria mainly in burnt
tissues. It acts by damaging cytoskeleton.
 P. aeruginosa strains that posses type III secretion
system are more virulent.
 3. Enzymes: Two enzymes
a. Elastase.
b. Alkaline Protease
 These(a &b)are toxic to tissues and facilitate the
invasion of organism into the blood stream.
 Other toxins include lecithinase and phospholipases.
These appear to act together to break down lipids and
lecithin.
 BIOCHEMICAL CHARECTERS

 Catalase-positive

 Oxidase-positive

 Nitrate reduction-positive
 Cont..
 Indole test-negative
 Methyl red test-negative
 Vp test-negative
 Citrate test-positive
 Urease test-negative
 Cont…
 Sugar fermentation tests
Glucose-only acid
Lactose-negative
Sucrose-negative
Mannitol-negative
 Pathogenesis:
 It mainly causes
1. Pneumonia.
2. UTI.
3. Sepsis in immunocompromized patients
4. Infections in burnt tissues.
 Pathogenecity is based on multiple virulence factors.
1. Endotoxins. Produce sepsis and septic shock.
2. Exotoxins. Two main exotoxins.
 PATHOGENESIS
 Important agent in causing nosocomial
infections
 Most common infections are

 Urinary tract infections following catheterisation

 Acute purulent meningitis following lumbar

puncture
 Post-tracheostomy pulmonary infection

 Septicaemia in debilitated patients

6. Also causes endocarditis in patients on I.V drugs use.
7. Severe external otitis ( malignant otittis externa) and
folliculitis in persons using bath tubs and swimming
pools.
8. Severe osteochondritis of foot
9. Corneal infections in contact lens users
 Mechanism Of Pathogenesis
 Caused by
exotoxins,proteases,elastases,haemolysins,lip
ases and enterotoxins
 Exotoxin A-lethal toxin

 Elastases-haemorrhagic lesions

 Enterotoxins-diarrhoeal disease

 Slime layer acts as a capsule and enhances

virulence
 Clinical findings:
It can cause infection any where in body but
commonly it causes
1. UTI
2. Pneumonia
3. Infection in burn wounds.
When disseminates it causes. --Sepsis
4. Necrotic skin lesions called eccthyma
gangrenosum.
 cont..
 Wound and burn infections
 Chronic otitis media and otitis externa

 Eye infections

 Acute necrotising vasculitis

 Infantile diarrhoea
 Resistance
 Susceptible to
heat,killed at 55°c in one hour
acids
beta glutarldehyde
Silver salts
strong phenolic disinfectants
 So,silver sulphonamide as a topical cream in
burns is used
 Antigenic structure
 O antigens
 Possesses 19 distinct,group specific O antigens
 Heat labile

 H antigens
 Two heat labile antigens recognised in
Ps.aeruginosa
 Cont…
 Resistant to
chemical agents
common antiseptics
disinfectants -ammonium compounds
chlroxylenol
hexachlorophane
antibiotics -aminoglycosides
cephalosporins
pencillins
 Typing methods
 Bacteriocin typing
 Three types of bacteriocins are produced-R,F,S

 Pyocin produced by test strain is employed to

assess the growth inhibition of 13
 Depending upon the growth inhibition of 13
indicators strains,105 types are recognised
 Most popular method used
 Cont..
 Phage typing
 Serotyping

based on O and H,17 serotypes of

Ps.aeruginosa are recognised
 Molecular Method
Restriction endonuclease typing with
pulsed-field gel electrophoresis(PAGE) is most
reliable
 Lab diagnosis:
1. Gram staining.
2. Inoculation of nutrient agar
3. Inoculation of selective medium like Macconkey’s
agar or EBM
4. Alkaline reaction on TSI agar
5. Oxidase test is positive
6. Fruity or grape like aroma
 Treatment:
 Resistant organism so treatment should be done
according to C/S
 However drugs of choice are Ticarcillin or Piperacillin
in combination with gentamicin or Amikacin