PSEUDOMONAS SPP.

Learning Outcomes
At the end of the lecture, student should be able to:
1. 1. Identify the species of Pseudomonas spp.
according to characteristic, Pathogenicity, and
habitat.
2. Identify the laboratory features of the
Pseudomonas spp.
3. Identify the appropriate treatment and antibiotic
can be used to kill the Pseudomonas spp.
Pseudomonas spp can be divided
more than140 species…..

• P. aeruginosa • P. mallei
• P. alcaligenes • P.pseudomallei
• P. chlororaphis (Burkholderia
pseudomallei)
• P. mendocina
• P.fluorescens
• P. monteilii
• P.putida
 Pseudomonas aeruginosa
• Characteristic
• Habitat
• Pathogenicity
• Virulence
 Characteristics
Asporogenous(not produce spore)
Resistant to high concentation of
salts and dyes.

Rod shape
 1-5 µm long
 0.5-1.0 µm wide obligate aerobe
Can growth in absence of O2 if
NO3 available

Motile
Active swimming by flagellum
Biofilm organism
Simple nutritient requirement
Gram Negative
Morphology different
 HABITAT
These pathogens are widespread in

Water
Animals

inhabiting soil

Human
Plants
 HABITAT

Pseudomonas aeruginosa can be found in water,

soil, sewage and vegetation

Frequently present in hospital environments (sinks,

bowls, drains, cleaning buckets and humidifiers).

Also can be found in intestinal tract.

Also can be found growing in eye drops, saline and

other aqueous solution.
 PATHOGENICITY

•An ubiquity organism, many infections

with Pseudomonas aeruginosa are
opportunistic hospital acquired, affecting
those are already in poor health and
immunosuppressed patients.
 Infections
Skin ●
Especially burn sites, wounds, pressure sores,
and ulcers.
Infection ●
Septicaemia may develop

Urinary ●
Usually following catheterization or
associated with chronic urinary disease.
infection
Respiratory ●
Especially in patients with cystic fibrosis or
conditions that cause immuno-suppression.
infections
External ear infection /
Eye infection
* often secondary to trauma or
surgery.
INFECTION
LABORATORY FEATURES:
Sample types
Morphology
Biochemical set
 MORPHOLOGY
Large, flat and greenish colonies (2-4 mm in diameter)
with irregular edges and typical metallic luster.
 The color is most visible on for instance TS-agar.
Sometimes, a clear hemolysis zone is obtained on blood
agar.
Has fruity smell (caramel, strawberry or raspberry
soda).
 Some strains produce a green fluorescent pigment,
pyoverdine.
Some strains can also produce a blue pigment,
pyocyanin.
LABORATORY FEATURES

Sample types

-Pus
-Urine
-Sputum
-Effusions
-Blood for culture
 Culture
 P. aeruginosa
- an obligatory aerobe.
- Usually recognized by the pigments it produces including :
 pyocyanin a blue-green pigment,
 pyoverdin (fluorescein) a yellow- green
fluorescent pigment.
- A minority of strains :
 non pigment producing.
 have a fruity smell due to the production of 2-
aminocetophenone.
 Grows over a wide temperature range 6-42°C with an
optimum of 35-37°C
 Blood agar
*produces large,flat,spreding colonies which are often
haemolytic and usually pigment- producing.
* Some strains produce small colonies or mucoid colonies
*The pigment diffuse into the medium giving it a dark
greenish-blue color.
*Some strains produce small colonies or mucoid colonies.
*If the culture is left at RT, the yellow-green of the
pyocyanin is intensified.
Mueller Hinton Blood
agar agar

Mac Conkey
agar
 Biochemical Reaction

TEST RESULT
Oxidase positive
Catalase positive
Motility positive
Indole negative
MR negative
VP positive
Citrate positive
Carbon Negative GLSM
TSI Red slope/Red butt
OF O and not F.
 Antimicrobial sensitivity

Resistant to most of the commonly

used antibiotics.
Antimicrobial that usually show activity
against Pseudomonas include:
• aminoglycosides,gentamycin
• polymyxin, imipenen and
fluroquinoles
Burkholderia pseudomallei
@
Pseudomonas pseudomallei
 Burkholderia pseudomallei @
Pseudomonas pseudomallei
• Formerly known as Gram negative rod with
bipolar staining ( like safety
pin )
• Small
• Motile

Fluorescence stain of Bulkholderia psuedomallei

 Pathogenicity
• Cause meliodosis ( pneumoenteritis )/Whitemore’s
disease in humans and animals.
• Latent infections may become active some years
later due to immunosuppression.
• Can be found in rice-growing areas. Mainly found
in Malaysia, Vietnam, Thailand, Myanmar, Guam
and Sri Lanka.
• Route of infection : enter open wounds but also
can through respiratory system.
 Specimens
• May be found in
 Sputum ( in pulmonary disease )
 Blood ( in systemic disease )
 Pus ( from abscesses )

• Highly infectious pathogen ( Hazard Risk

Group 3 )
 Culture
 An aerobe.
 Produce non-hemolytic, small, dry, ringed and striated
colonies on blood agar after overnight incubation.
 The colonies become wrinkled after several days
incubation
 Culture
• On MacConkey agar, it forms acid from
lactose. Colonies have a rough and corrugated
appearance.
• Does not produce pyocyanin or fluorescein
but forms an orange-
brown pigment.
 Culture
• Give off an ammoniacal smell.
• A pellicle (skin) is formed on broth cultures.
 Biochemical tests

• Oxidase positive
• Forms acid (oxidatively) from glucose, lactose,
maltose and mannitol.
• It also grows at 42°C.
• Cultures should be sent to a Reference Microbiology
Laboratory for serological confirmation.
 References
• Kenneth Todar , 2009. Opputunistic infection of Pseudomas
aeruginosa .Retrieved on 8 January 2013 from
http://textbookofbacteriology.net/themicrobialworld/Pseudom
onas.html
• Micheal Jenkins, 2006. Pseudomonas pseudomallei. Retreived
on 8 January 2013 from
http://web.mst.edu/~microbio/BIO221_2006/P_pseudomallei.h
tm
• John P.Cunha and William C. Shiel , 2013. Melioidosis. Retrieved
on 9 January 2013 from
http://www.medicinenet.com/melioidosis/article.htm
• Allen C.Cheng and Bart J. Currie,2005. Melioidosis:
Epidemiology, Pathophysiology and Management. Clin Microbial
Reviews.383-416.