Pseudomonas aeruginosa

About Pseudomonas aeruginosa

gram-negative aerobe bacteria
Commonly found in the environment
At any moist location

Common cause of nosocomial infections

P. aeruginosa is an opportunistic pathogen

Extremely broad host spectrum
Hardly any infections in the normal human host
Severe immunodeficiencies and medical devices
predispose the patients to P. aeruginosa infections
Broad spectrum of clinical symptoms

Urinary tract infections

Pulmonary infections
Soft tissue infections
Sepsis
Bone and joint infections
Endocarditis

P. aeruginosa infections
P. aeruginosa infections are of particular
concern for Cystic fibrosis patients
Burn patients
Hospitalised patients
Case mortality rate for patients infected with P.
aeruginosa approaches 50%

Nosocomial infections
Fourth most common isolated nosocomial
pathogen accounting for approx.
10 % of all hospital acquired infections.
Patient-to-patient spread and direct patient contact
with environmental reservoirs

disinfectants,
respiratory equipment,
food,
sinks, taps

Diagnosis of P. aeruginosa
Isolation and lab identification of the pathogen
P. aeruginosa grows well on most laboratory media
Identified on the basis of its:

Gram morphology,
inability to ferment lactose,
a positive oxidase reaction,
its characteristic odor,
its ability to grow at 42 C.
Fluorescence is helpful in early identification of P.
aeruginosa colonies and may also help identify its presence
in wounds.

Treatment of P. aeruginosa infections

P. aeruginosa is frequently resistant to
many commonly used antibiotics.
To archive synergy a combination of e.g.
gentamicin and carbenicillin is frequently
used.
No vaccines so far

Pathomechanisms
Adhesion
Pili, flagella and fimbriae

Invasion
Extracellular enzymes and toxins (proteases, elastase,
phospholipases, rhamnolipids, Exotoxin A)

Dissemination
Leukocidin inhibits neutrophils und leukocytes
LPS (Endotoxin)

Protection
Capsule (Alginate)

Interbacterial Communication

Quorum Sensing in P. aeruginosa

McKnight et al, 2000

....It is time to close the book on infectious

disease.
William Stewart, Surgeon in a
message to the United State
Congress in 1969

Bacterial Biofilms

Biofilms in the environment

Catheter associated biofilms

Chronic biofilm infections

Despite even intensified antibiotic therapy,

no eradication of chronic P. aeruginosa
infections of the cystic fibrosis lung

Cystic fibrosis
Most common life-threatening inherited genetic disorder in
the Caucasian population
Mutation in the cystic fibrosis transmembrane conductance
regulator (CFTR) gene
one in every 25 carry the mutated recessive gene and more
than 1 in 4000 live births suffer from CF.
Life expectancy:
Until the 1930s: the life expectancy of a baby with CF was only a few months,
in the 1980s, most deaths from CF occurred in children and teenagers. Today with
improved treatments, nearly 40 percent of the CF population is aged 18 and older,
for a person with CF the median age of survival is nearly 37 years.

Cystic Fibrosis affects a number of organs in the body, cycles

of infection and inflammation lead to a progressive
deterioration of lung function.

Chronic infection of the

Cystic Fibrosis lung

CF and Transplantation Centre

360 patients regularyly attend the CF outpatient clinic at

the Medical School Hannover, Germany

Informations of more than 500 CF patients in the local CF

register

2000 2005: 100 lung transplantation / year at the MHH

Indications for lung transplantation

Lung Transplantation in Germany

Post lung transplant mortality

Chronic infectious diseases

Slow progressing infections

Functional loss of the affected organ
Highly resistant to host defences and antimicrobial therapy
Recovery is rare
Role in the development of cancer, peptic ulcer and
possibly atherosclerosis

Why is traditional antimicrobial therapy

ineffective against biofilm bacteria?

Biofilms, City of microbes

Adaptation and survival is facilitated by diversity

Emergence of morphological distinct nichespecific phenotypes

From: Rainey R, Travisano M. Nature, 1998; 39: 69-72

Phenotypic Diversity via the selection of nichespecialists

Small colony variants (SCV)

SCVs of P. aeruginosa in CF
Slow growing subpopulations (3% of the P. aeruginosa
positive sputum specimens)
SCVs exhibit an increased resistance towards a broad
spectrum of antimicrobial agents
The recovery of SCV correlates with parameters revealing
poor lung function and an inhalative antimicrobial therapy
Fast growing revertants can be isolated from the SCV
population

Auto-aggregation in liquid cultures

CupA encoded fimbria expression in P. aeruginosa

M. Rohde, GBF Braunschweig

Molecular Mechanisms controlling the

conversion to a SCV biofilm phenotype

Biofilms, City of microbes

Cooperative traits and coordinated behavior

of bacteria

Quorum Sensing in P. aeruginosa

McKnight et al, 2000

Pseudomonas Quinolone Signal (PQS)

Hydroxy Alkyl Quinolones

PQS

HHQ

PQS directly interacts with bacterial DNA in a

computed model

PQS enhances iron dependent DNA fragmentation

Control
DNA

DNA

DNA
+ PQS

DNA
+ HHQ

DNA
+ Fe(II)

DNA
+ PQS
+ Fe(II)

DNA
+ HHQ
+ Fe(II)

PQS enhances the fragmentation of DNA in vivo

PQS production is linked to the release of DNA

PAO1 wild-type

PQS non-producing mutant

Regulated DNA release in response to

environmental stresses

DNA breakdown products enhance PQS

production

Autoinductive PQS production cycle

PQS

PQS

Rhl
Quorum Sensing

pqsA-E transcription

Lectin
Cell death

DNA

Pyocyanin
Virulence factors
Biofilm formation

Phenotypic Diversity is PQS dependent

P. aeruginosa WT

P. aeruginosa PQS negative Mutante

Establishment of a chronic state of infection

Biofilm
Formation

Bacterial
Diversity
Chronic
Infections

Interbacterial
Communication

Molecular mechanisms underlying genetic

diversity
Horizontal gene transfer
Hypermutation
Adaptive mutations

Tiling Arrays

Microarray hybridisation-based method technique to find

mutations in bacterial genomes was used to study
metronidazole resistance in Helicobacter pylori
TJ Albert et al. Mutation discovery in bacterial genomes: metronidazole
resistance in Helicobacter pylori. Nature Methods 2, 951 - 953 (2005).

Thank You for your attention!

Systems Biology
Prediction of the
impact of cellular
metabolism on the P.
aeruginosa quorum
sensing mediated
virulence phenotype
Use a theoretical model to complete
the knowledge on the system, to plan
new experiments and to predict the
behavior of the system under changing
genetic or environmental conditions