Transcript Psudomonas putida and fluorecences Prepared by: Ghada Rayyan D

Psudomonas putida and
P.fluorescens
Prepared by: Ghada Rayyan
Dr. Abdelraouf A. Elmanama
Classification
Higher order taxa:
Bacteria;
Proteobacteria;
Gammaproteobacteria;
Pseudomonadales;
Pseudomonadaceae.
Pseudomonas aeruginosa group
Pseudomonas chloroaphis group
Pseudomonas fluorescens
Pseudomonas pertucinogena group
Pseudomonas putida group
Pseudomonas stutzeri group
Pseudomonas syringae group
Laboratory Diagnosis.
Gram-negative.
Rod-shape.
Aerobic metabolism.
Fluorescent pseudomonads.
Motile by tow or more polar flagella.
Able to grow on a wide variety of organic
substanse.
Produce a slime layer that cannot be phagocytosed.
Production of surface- colonising biofilms.
Most pseudomonads are free-living saprophytic
organism in soil or water.
They play an important role in decomposition,
biodegradation, and the carbon and nitrogen
cycles.
Pseudomonads are characterised by great
metabolic diversity and are able to utilize a wide
range of carbon sources.
They are important organism in bioremediation.
Pseudomonas are able to grow in unexpected
places.
They have been found in areas where a lot of
pharmaceuticals are prepared.
Any carbon source suchas soap.
Antiseptics such as ammonium compounds
and bottled mineral water.
This ability to thrive in harsh conditions is aresult
of their hearty cell wall that contains porins.
Cell structure and metabolism.
they "grow well in mineral salts media
supplemented with any of a large number of
carbon sources" .
Some researches seek to exploit P. fluorescens to
partially or completely degrade pollutants such as
styrene, TNT, and polycyclic aromatic
hydrocarbons.
Several strains of this bacteria also have the ability
to suppress plant diseases by "protecting the
seeds and roots from fungal infection“.
This ability is due to secondary metabolites
produced by these bacteria such as antibiotics,
siderophores, and hydrogen cyanide as well as the
ability of these bacteria to rapidly colonize the
rhizosphere and out-compete some of pathogens.
Pseudomonas putida
Gram-negative bacteria with multitrichous
flagella.
P.putida is a unique soil microorganism,
which can resist the adverse effects of
organic solvents.
Also is capable of decontaminating organic
substances including solvents, such as
toluene, one of the components of gasoline.
It is an example of an organism that can be
used to carry out bioremediation.
Polluted soil and groundwater purification
technology which uses the power of
microbes, renders many toxic organic
substances harmless.
Magnetite-immobilized cells of Pseudomonas
putida 5-X can be used as biosorbent in
copper removal from aqueous solutions.
Pseudomonas putida is listed among
microorganisms most commonly found in
various environments such as various
consumer products including paints and
solvents.
It is also listed as one of safe and effective
pesticidal microorganisms.
Pseudomonas fluorescens
(motile rods associated with food spoilage)
Tow phenotyping characteristicsof p.fluorescens
that distinguish it from p.putida are:
it is ability to grow at 4 C
AND it is ability to hydrolyze gelatin.
These characteristics help explain its frequent
involvement in spoilage of refrigerated food, in
particular chicken and processed meats. If it's
fluorescent, get rid of it!
clinical manifestations.
Bacteria in the P.fluorescens-P. putida complex
have been isolated from lizards, insects and
mammals.
Clinical sources from which strains of these species
have been isolated include respiratory tract
specimens, pleural fluid, urine, cerebrospinal fluid,
feces, blood, and a variety of other materials.
p.Putida thrives in moist enviroments.
Its isolation from clinical specimens, especially
non-sterile sites,and is considered to have
uncertain pathogenic.
More than half of the clinical isolates of
Pseudomonas bacteria produce pyocyanin, a bluegreen pigment.
The Pseudomonas fluorescens group are
nonpathogenic saprophytes that also produce a
pigment, particularly under conditions of low iron
availability.
This pigment is a soluble, greenish, fluorescent
pigment that led to the group's name.
These bacteria are generally obligate aerobes;
however, some strains can utilize NO3 instead of O2
as an electron acceptor.
The taxonomy of p.fluorescens and
P.putida
The first works on the taxonomy of P. fluorescens
and P. putida were based on phenotypic characters
such as metabolic tests, fatty acid composition and
protein profiles.
These studies led to the subdivision of P.
fluorescens and P. putida into 5 and 2 biovars,
respectively.
genotypic characters based on the variability of
consensus genes (rDNA) or of the total genome
(DNA-DNA hybridization).
combining both recent phenotypic and genotypic
methods, is required to clarify the taxonomy of P.
fluorescens and P. putida.
Pseudomonas putida is a ubiquitous soil
bacterium that has significant potential for
bioremediation of numerous compounds.
P. putida genome sequence will identify the real
potential of this organism in various
biotechnological areas including the production of
natural compounds, and remediation of polluted
habitats.
Pathogenesis
Heat stable lipases and proteases are produced by
p.fluorescens and other similar pseudomonads,
and cause milk defects such as bitterness,
rancidity, fruity and cardboardy flavor, casein
breakdown, and ropiness due to production of
slime and coagulation of proteins.
The main property that becoming important
opportunistic pathogens is their inability to grow at
body temperature.
they are rarely pathogenic for humans, even
though they have been found associated with
empyema, urinary tract infections, septicemia, and
various other episodes.
Although their virulence may be low, P .putida and
P .fluorescens should be considered as potentially
pathogenic.
In immunocompromised patients p.putida
has been found to cause septicemia and
septic arthritis.
Nosocomial transmition of p.putida has only
reported twice.
Strains of p.fluorescens have been frequently
identified as contaminant on the skin of humans
and as agent causing pseudobacteremia and
procedure- related infection in hospitalized
patients and infections resulting from transfusion
with contaminated blood components.
Biochemical characteristics
A rapid nitrate test was found to be useful in
distinguishing P. aeruginosa (positive) from P.
fluorescens and P. putida (both negative).
A shortened gelatin test differentiated P.
fluorescens (positive) from P. putida (negative).
Produced fluorescin but no pyocyanin or pyorubrin.
Oxidase test – positive.
10% lactose - both negative.
Oxidation of dextrose - both positive.
Oxidation of maltose - both negative.
Nitrate reduction -both negative.
Gelatin liquefaction – p.fluorescens positive,
P.putida negative.
Growth on cetramide – positive.
Are growth at 4 C AND at 35 C unlike p.aeruginosa
able to growth at 42 C.
CHROM agar:
Pseudomonas is a chromogenic media, for
isolation of specimens, giving detection of
Pseudomonas sp. by colony colour.
Pseudomonas sp. develop as easily distinguishable
blue-green coloured colonies, clearly visible under
normal lighting conditions. Other bacterial species
are inhibited or give colourless colonies.
CHROMagar Pseudomonas
Pseudosel Agar
Pseudosel Agar is the BBL medium of choice for
the selective isolation and identification of
Pseudomonas aeruginosa.
It is a modification of King's Tech Agar, stimulating
enhanced pyocyanin production by Pseudomonas
while inhibiting other organisms with centrimide.
Pseudosel Agar also detects fluorescin produced
by some pseudomonads.
Pseudosel Agar
Pseudomonas agar
Pseudomonas agar F. Flo Agar
Pseudomonas agar P.Tech Agar
pseudomonas agar F, also as Flo Agar, is used for
the enhancement of fluorescin production and
Pseudomonas agar P, also known as Tech Agar, is
used for the inhancement of pyocyanin production
by pseudomonas.
Antibiotic sensitivity test
p.fluorescens and p.putida were very sensitive to
kanamycin.
And p.fluorescens and p.putida were resistant to
carbenicillin.