Quorum sensing Vibrio harveyi

Three signals
integrated in
parallel
At low
concentrations the
signal sensors
work as
phosphatases
At high
concentrations the
signal sensors
work as kinases

Pseudomonas aeruginosa Bacillus subtilis

Two signals Two signals
integrated in regulate two
series diferente fates
Acumulation of ComX -
LasI increases its competence
production and
also that of RhlI CSF - sporulation

Sequencial
expression of two
sets of genes
Quorum sensing decides individual cell fate in large
populations

Streptococcus pneumoniae Quorum sensing

Quorum sensing regulates a Response to the host
coordenated population
response limited in time A. tumefaciens – Presence of opines lead to the
production of TraR (LuxR)

Response without a signal

Species have luxR homologues without luxS

Quenching of the signal

B. subtilis – AiiA cuts the lactone ring
Variovorax paradoxus – Acilase cuts the lactone ring
Delisea pulchra (read algae) – furanone degrades
LuxS (avoids colonization by bacterial biofilms)

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Quorum sensing Quorum sensing
Regulate: What is the role of quórum sensing?
Alternative diferentiation pathways Count the number of individuals in a population
Sporulation
Competence
Swarming

Coordination
Competence
Antibiotic production
Virulence factor expression
Gene expression (polysaccharides, flagella)

Quorum sensing Quorum sensing

Count the number of individuals in a population
Synthesis of the sensor constant and
independente of the context
Difusion of the sensor to the outsider and in the
external millieu constant
Persistence of the signal in the external millieu
constant
What is the role of quorum sensing?
Count the number of individuals in a population
Coordinate the expression of genes in the population
(independent of the number of individuals)
Activate a specific response in the face of
certain stimuli
Avoid complex responses when
replication is the most successful
strategy

Planktonic life Life in a biofilm

In vitro studies Cells adherent to a
surface
Unicelular life
Encased in a
polymeric material
Biofilm
The majority of bacterial life in vivo Composed of one or more species
Life in a consortium (in society) Ex: dental plaque

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 Life in a biofilm Life in a biofilm

Life in a biofilm Life in a biofilm

Surface conditioning

Accumulation of the nutrients and

particles from the medium on the surface
Polisaccharides
Proteins

Zone of higher osmolarity

Life in a biofilm Life in a biofilm

Reversible adhesion
Random colision with the
surface due to turbulence

Active movement (ex: flagella,

fimbriae)
Quimiotactic
Substrate – natural (tooth) ou artificial (catheter) Hydrophobic interactions

Fimbriae, pili

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Life in a biofilm Life in a biofilm
Irreversible adhesion Formation of micro-colonies
Expression of a specific Celular proliferation
genetic program (?)
Redistribution of cells on the
Surface proteins surface (motility)
Extracelular polysaccharides twitching
Condensation of groups into micro-
Formation of biofilms occurs more
colonies
readily in turbulent fluids
Cell division
More productive collisions with surfaces?
Recruitment of plaktonic cells from the liquid
medium

Vida de um biofilme Life in a biofilm

Maturação do biofilme Dispersion
Expressão de um programa Separation of cells that have
específico (?) reverted to a planktonic
Síntese da matriz program
extracelular polimérica Separation of mature biofilme
(EPS) structures
Manutenção de uma
arquitectura tridimensional Change in the behaviour of cells in regions
of the biofilme that lead to the detachment of
Heterogeneidade metabólica (activação de adjoining regions
programas genéticos específicos?)

Biofilm structure Biofilm structure

Irregular structure

Extracellular polymeric matrix

(85% of total volume)
Polysacharides, DNA, proteins
viscoelastic (variable) Influence of physical factors on structure

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 Biofilm structure Biofilm structure

Access to liquid phase (nutrients, gases) Structuring generates microenvironments

influences biofilme architecture Activation of different genetic programs

Biofilm structure Biofilm structure

Interaction between different populations Mutants lasI and rhlI in P. aeruginosa
within the biofilm
Metabolic – possibly different species

Signalling –
quorum sensing
systems

Dispersion Dispersion
Resulting from the action of external forces on Activation of an alternative genetic program
the biofilm (due to stress inside the biofilm)

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 Dispersion Definition of a biofilm
Activation of an alternative genetic program Fixed microbial community
(due to stress inside the biofilm)
Cells irreversibly attached to a surface or
Quorum
interface
sensing
Cells encased in a matrix of extracellular
polymeric substances (EPS)
Cells exhibit altered phenotypes and
growth due to the expression of a distinct
genetic program

Biofilm properties Biofilm infections

Increases resistance to antimicrobials and
disinfectants
1) Difficulty in penetrating the matrix
2) Growth rate of microorganisms
3) Physiological changes due to biofilm growth
More resistance to the host immune system
Endocarditis – colonization of a damaged heart
valve (vegetation)
Cystic fibrosis – colonization of the lung due to a
congenic disease.
Periodontitis – colonization of the periodont
Chronic bacterial prostatitis– Colonization of the
prostate ducts

Biofilm infections Avoid biofilme colonization

Colonization of foreign bodies Three pharmacological targets
Catheters Bacterial sensors regulating chemotaxis
Contact lenses
Endotracheal tubes Two-component regulators
Intra-uterine devices (IUD)
Cell structures responsible for the
Prosthetic devices (cardiac valves, joints) interaction with the surface
Pacemakers